identifier
stringlengths 11
32
| pdf_url
stringlengths 17
4.62k
⌀ | lang
stringclasses 120
values | error
stringclasses 1
value | title
stringlengths 2
500
⌀ | source_name
stringlengths 1
435
⌀ | publication_year
float64 1.9k
2.02k
| license
stringclasses 3
values | word_count
int64 0
1.64M
| text
stringlengths 1
9.75M
|
|---|---|---|---|---|---|---|---|---|---|
https://openalex.org/W1834315828
|
https://www.biodiversitylibrary.org/itempdf/37678
|
English
| null |
First report on economic zoology,
| null | 1,903
|
public-domain
| 95,108
|
56
^^^^ LONDON:
PRINTED
BY ORDER
OF THE
TRUSTEES. SOLD BY
Longmans &
Co., 39, Paternoster Eow, E.G.; B. Quaeitch, 15, Piccadilly, W,
DuLAU & Co.,
37, Soho
Square, W. ; Kegan Paul, Trench,
Trubner &
Co., Charing Cross Road, W.C. ;
and at the
British Museum (Natural History), Cromwell Road, S.W. fttU ERRATA. Page iv. line 16 for moritans read morsitans. Page XV. line 30 for morrhuae read morrhua. Page 5 line 21 for Jensenii read Jensen. Page 6 line 15 for laying read lying. Page 36
after line 5 add "Dilute with 30 times
its bulk
of water before use." of water before use." Page 76 line 8 for Jig. 9,
c, read Jig. 8,
c. Page 77 line 6 for on read as. Page 77 line 6 for on read as. Page 87 line 2 for p. 115, read p. 105. Page 105 line 28 for soluble read insoluble. Page 127 line 36 for sulphate read sulphite. Page 159 line 31 for fatile read fatale. Page 169 line 20 for occompamjing read accompanying. PBEFACE. The present volume consists primarily of a
series of Reports
to
the
Board of Agriculture, of Eeports and letters to a variety of unofficial
correspondents,
and
of Reports
to
the
Foreign
Office and
to
the
Colonial
Office, drawn up by Mr. F. V. Theobald during the years
1901-1902. Mr. Theobald
has
recently
been
employed
by
the
Trustees of the British Museum to assist the Director in such work,
especially with a view to furnishing the Board of Agriculture with
scientific information on Economic
Zoology,
in
accordance with a
request made by
that
Department
of
his
Majesty's Government. Mr. Theobald is well known
as an authority on Economic Zoology,
and has
the advantage,
in carrying
out his
work
at
the Natural
History Museum,
of consulting with the various
specialists on
the
scientific staff, as well as of making use of the collections and library
of the Museum. I have added
to Mr. Theobald's Reports an introductory scheme
or outline of the study known as Economic Zoology, in the form of
a
classified survey
of the various
sub-divisions which
it
is found
convenient to recognise in the treatment of this
subject. For
this
classification
I am responsible, whilst Mr. Theobald has been good
•enough
to
fill in the
list of selected examples. I have
also added
some
correspondence
on
Tsetse-fly
disease, and
on
the
proposed
investigation of the Pearl Fisheries
of Ceylon, and on the Marine
Resources
of the West Indies, which
arose from my being consulted
in my
official
capacity
by
his Majesty's
Secretaries
of State
for
Foreign Affairs and for the Colonies. The Trustees have ordered the publication
of the present Report,
in
order that
the valuable information which
it
contains may be
made
easily
accessible
; and
further,
in
order
to make
clear
the
nature and amount of scientific information on matters
of economic
importance which the
staff of the Zoological Department
is almost
daily called upon and
is prepared to furnish
to the public service or
to individuals. It must be remembered that the Reports and letters
printed in this volume form only a portion
of the work of economic
importance which
is carried out by this Museum, in addition
to the First Report on Economic Zoology. iv far more extensive work
in
the pure
science
of Natural
History,
which
is
the primary occupation of
its
official
staff. British Museum (Natural History),
London, S.W. PBEFACE. The Trustees
published
in
1901
a
descriptive
treatise
on
Mosquitoes
in
three
volumes, with forty-two plates, which was prepared by Mr. Theobald
in connection with the specimens of Culicidfe ah'eady in the Museum,
and others specially collected
for the work, with a view
to assisting
in the
study
of the relationship of Culicidse to Malaria and
other
diseases. A supplementary volume of this work, by Mr. Theobald,
has been completed and published in the present year. Also
in the
present year the Trustees have published an
illustrated monograph
on
the
Tsetse-flies,
by
Mr. Austen,
Assistant
in
the
Zoological
Department. Our
rapidly increasing knowledge of the
activity of
the minute parasites known
as Trypanosoma, as the specific causes
of disease both in man and in horses and cattle, renders an accurate
knowledge of the species of Tsetse-flies necessary, since one of these
flies,
the
Glossina
moritans
of
Westwood,
is
the
carrier
of
the
Trypanosoma causing the deadly disease of horses and cattle known
in South Africa as Nagana, and
it is possible that
other species of
Glossina
are
concerned,
in
a
similar way,
in
the
distribution of
disease. It is not, however, only in correspondence and publications, and
in the researches of the
naturalists of the
staff that
this Museum
renders direct assistance
to the development of the knowledge and
application of Economic Zoology. The large study collections of the
JMuseum have, for a long time past, comprised important series from
all parts
of the world of carefully named and recorded specimens
of
animals having economic importance, either as pests or as sources
of
commercial products. In addition
to
these,
several
cases
are now
exhibited in the North Hall of the Museum, in which the life-bistory
and activities of animals important to man in one or other of the
relations recognised in the
classification adopted in
this volume, are
illustrated with
a view
to
the
edification
of
the
public, and
the
promotion
of the public
interest in the thorough scientific treatment
of the subject. I have
to
thank
the
Board
of
Agriculture
for
permission
to
reproduce some of the Keports furnished to the Board. E. EAY LANKESTEE. British Museum (Natural History),
London, S.W. May 15th, 1903. May 15th, 1903. G R
u r
E. Feuit Pests
: An
enquiry
re
Gall Mites
in
Black-currant
bushes; The
Apple-Blossom
Weevil ; Strawberry Beetles
; Slug-worms on Fruit trees and Hedgerows
;
Maggots in Apples
; Maggots in Imported Apples
;
Infestation of Fruit
trees by Winter Moth caterpillars
; The Pear Midge {Diplosis jiyrivora)
;
Scale
disease and "false scale" amongst
fruit
trees and bushes; Scale
Insects on Plum Trees
; The Apple Bark-louse or Mussel Scale (Mytilaspis
pornoritm)
; Eggs on Apple
trees, and a further remedy for Mussel Scale
;
Aphides (A.mali) on Apple trees
; The Peach Aphis
; A general wash for
Fruit
trees
; Further
Information
re Winter Washing of Fruit trees
Canker fungus (Nedria ditissima) on Apple twigs mistaken
for insect
work
; Enquiry as to poison for Moles ...... 3 G R
u r
E. SiB-GBOUP
i?.—Section
I. ANIMALS INJURIOUS TO AGRICULTURE. SiB-GBOUP
i?.—Section
I. ANIMALS INJURIOUS TO AGRICULTURE. I'ereal Pests: Eel-worm
disease
in
oats; The
Frit Fly (Oscinis frit) on
oats
; Smut
in Bailey and Insects. Root Crop Pests
:
SilpTia rugosa on
Turnips,
and
other
Silphidaj;
Notes
on
Surface
Larvre; The Pigmy
Mangold
Beetle (Atomaria
linearis); Black
Fly on
Mangolds (Aphis
atriplids,
Linn.);
Flies
(Bibionidas)
on
Mangolds;
Muscid
Larvfe
attacking
roots
;
Chafer
Larvas
(Melolonthidx)
;
Leather
Jackets
(Tipulidai)
; The
Green
Rose- Chafer
on
Beans
and
Currant
Bushes. Potato Pests
: Myriapoda in Potatoes ; Wire-worm (Agriotes murinus)
in Potatoes; A new Potato Feeder, the Cinnabar Moth (Euchelia jacobem). Mustard Pests
:
Destruction of the Mustard Beetle. Feuit Pests
: An
enquiry
re
Gall Mites
in
Black-currant
bushes; The
Apple-Blossom
Weevil ; Strawberry Beetles
; Slug-worms on Fruit trees and Hedgerows
;
Maggots in Apples
; Maggots in Imported Apples
;
Infestation of Fruit
trees by Winter Moth caterpillars
; The Pear Midge {Diplosis jiyrivora)
;
Scale
disease and "false scale" amongst
fruit
trees and bushes; Scale
Insects on Plum Trees
; The Apple Bark-louse or Mussel Scale (Mytilaspis
pornoritm)
; Eggs on Apple
trees, and a further remedy for Mussel Scale
;
Aphides (A.mali) on Apple trees
; The Peach Aphis
; A general wash for
Fruit
trees
; Further
Information
re Winter Washing of Fruit trees
Canker fungus (Nedria ditissima) on Apple twigs mistaken
for insect
work
; Enquiry as to poison for Moles ...... 3 SiB-GBOUP
i?.—Section
I. ANIMALS INJURIOUS TO AGRICULTURE. I'ereal Pests: Eel-worm
disease
in
oats; The
Frit Fly (Oscinis frit) on
oats
; Smut
in Bailey and Insects. Root Crop Pests
:
SilpTia rugosa on
Turnips,
and
other
Silphidaj;
Notes
on
Surface
Larvre; The Pigmy
Mangold
Beetle (Atomaria
linearis); Black
Fly on
Mangolds (Aphis
atriplids,
Linn.);
Flies
(Bibionidas)
on
Mangolds;
Muscid
Larvfe
attacking
roots
;
Chafer
Larvas
(Melolonthidx)
;
Leather
Jackets
(Tipulidai)
; The
Green
Rose- Chafer
on
Beans
and
Currant
Bushes. Potato Pests
: Myriapoda in Potatoes ; Wire-worm (Agriotes murinus)
in Potatoes; A new Potato Feeder, the Cinnabar Moth (Euchelia jacobem). Mustard Pests
:
Destruction of the Mustard Beetle. CONTENTS. PREFACE
iii
INTRODUCTION. The Classification of Anoials from the
Point of View of EcoNOiiic Zoology
. . . . xi ub-group
J5.—Section
II.
ANIMALS
INJURIOUS TO HORTICUL-
TURE. Sub-group
J5.—Section
II. ANIMALS
INJURIOUS TO HORTICUL-
TURE. Land Bugs on Chrysanthemums (Lygus pratensis)
; The destruction of Ants
The destruction of subterranean
insects and other ground garden pests
Directions for the employment of the Gas Treatment under glass
; White
Grubs or Maggots {Fhorhia bmssicai) causing
great damage amongst
cabbage, carrots and broccoli, and exterminating Cabbage-fly
,
. 30-36 First Report on Economic Zoology. vi PAGES
Sub-group
J?.—Section
III. ANIMALS INJURIOUS' TO FORESTRY. Willow-beetle
at
Norwicli
(^Sarperda
carcharias, Linn.);
Sirex
Flies
in
Fir
trees
{S. juvenms
and
8. gigas); The Poplar Saw-fly
{Cladius
viminalia)
; Saw-fly
Larvre on Willows ;
Insect Galls on Osier plants
The Felted Beech-coccus (jCryptococcus fagi)
;
Chermes corticalis on Pine
trees
36-40 Appendix. Tapeworm in Sheep at Okehampton
; Black Wire-worm in Mangolds
;
List of
leaflets prepared and revised for the Board ..... 49-50 Tapeworm in Sheep at Okehampton
; Black Wire-worm in Mangolds
;
List of
leaflets prepared and revised for the Board ..... 49-50 Tapeworm in Sheep at Okehampton
; Black Wire-worm in Mangolds
;
List of
leaflets prepared and revised for the Board ..... 49-50 GROUP
B. Origin and Varieties of Domesticated Geese ...... 53-54r GEO UP
F. The Furniture Beetles and Clothes Moths
;
Insects and Mites
in Furniture
the Larder Beetle (Dermestes lardarius) ; Weevils amongst stored com
(Calandra granaria) ......... 41-4:7
Fungoid disease in Black-currant leaves (Septoria ribis)
; Gooseberry fungus
(^Puccinia pringsheimiana) ........ 47-49 GEOUP
D. Dipterous larva? in human excreta
; Correspondence on the Mosquito annoy-
ance at Blackheath ......... 55-59 GROUP
E. Contents. vii The Pear Leaf Blister Mite
; Big Bud in Currants
;
Oribatidse or Beetle
Mites
;
Hover
Flies
or Syrphidm
; The
Mussel
Scale
(Mytilaspis
pomormn). Hop Pests
: Woodlice in Hops. Cereal Pests : Beetles
in Barley affected with Smut. Potato Pests
: The Rosy Rustic Moth
(Hydnecia micacea)
; Surface larvas attacking Potatoes and Celery ; The
Pigmy Potato Beetle (Bathyscia
wollastoni)
;
Millepedes
attacking
Potatoes ; Notes on the Colorado Beetle in England. Root-crop Pests
:
Cabbage Aphis on Turnips
;
Tipulidse or Daddy-long-legs
. . 62-104: Section
III.
ANIMALS INJURIOUS TO FORESTRY. Section
III. ANIMALS INJURIOUS TO FORESTRY. Goat-Moth larvse attacking Willows
; Insects on Osiers and Willows ; Insects
on Elm and Willow ; Pissodes notatus ravaging Austrian Pines
; The
Spruce Gall Aphis (Ohermes ahietis) ...... 113-119 Goat-Moth larvse attacking Willows
; Insects on Osiers and Willows ; Insects
on Elm and Willow ; Pissodes notatus ravaging Austrian Pines
; The
Spruce Gall Aphis (Ohermes ahietis) ...... 113-119 Sub-group
G. The Cigar Beetle {Lasioderma
testaceum. Duff)
; The Indian Meal Moth
(Flodia inttrpundella) attacking Almonds ; The Larder Beetle
. 125-127 SUB-GBOUP A. Acarine
household
pests
(^Glyciphagus
domesticus
and
G. spinipes)
;
Anohium
tcsselJatum
in
St. Alban's
Cathedral
;
Earwigs
causing
annoyance indoors ......... 119-124 GENERAL SUBJECTS. Green Matter in Lewes Public Baths ;
Short Reports .... 127-129 Green Matter in Lewes Public Baths ;
Short Reports .... 127-129 Section
II.
ANIMALS INJURIOUS TO HORTICULTURE. Julidx destroying plants in the gardens of Downton Castle, Ludlow
; Wood-
lice in Gardens
; A new Phytoptid Disease in Violas ; The Narcissus Fly
{Merodoii
equestris)
; The Marguerite Fly and its Destruction
;
The
Carrot-fly and Aphides on
Carrots
; Correspondence and
Report on
Insects in Orchid Houses
; Fumigation for Mealy Bug
. . . 105-113 Julidx destroying plants in the gardens of Downton Castle, Ludlow
; Wood-
lice in Gardens
; A new Phytoptid Disease in Violas ; The Narcissus Fly
{Merodoii
equestris)
; The Marguerite Fly and its Destruction
;
The
Carrot-fly and Aphides on
Carrots
; Correspondence and
Report on
Insects in Orchid Houses
; Fumigation for Mealy Bug
. . . 105-113 Sub-group A. Teredo and Canadian Timber ........ 143-144
• Teredo and Canadian Timber ........ 143-144
Other Short Reports
..-.•. •. . <. . 144-145 GROUP
D. A poisonous Land Bug in Singapore ; The Screw Worm in St. Lucia
. 130-131 First Report on Economic Zoology. viii GROUP
E. iSuB-GBOUP A. PAGES
The Screw Worm in Cattle at St. Lucia
; Pony Flies .... 132-133
Stfe-GBOUP B. Scale InsectB in Monte Video {Mytiiaspis citricola)
; Scale on Pineapples
in Jamaica {Diaspis bromelise)
; Pine
Beetle (Hylesinus jnniperda);
Damaged
Coffee-berries
from
Uganda
and
Costa
Rica
;
Weevils
( Uyjaomeces sqitamosk^) defoliatihg riibVier ; IiiseCt P^sts 'of Wfest Indies 5
other Reports
. . . 133-142 Stfe-GBOUP B. Scale InsectB in Monte Video {Mytiiaspis citricola)
; Scale on Pineapples
in Jamaica {Diaspis bromelise)
; Pine
Beetle (Hylesinus jnniperda);
Damaged
Coffee-berries
from
Uganda
and
Costa
Rica
;
Weevils
( Uyjaomeces sqitamosk^) defoliatihg riibVier ; IiiseCt P^sts 'of Wfest Indies 5
other Reports
. . . 133-142 A. REPORTS TO THE FOREIGN OFFICE. I'setse-fly and its Connection with the Buffalo (Sos caffa) (Correspondence) ; 147-155
White Ants or Termites in the Sudan (Correspondence and Report)
; 155-163
Locust Plagues in the Sudan (Correspondence and Report)
. . 163-169
B. REPORTS TO THE COLONIAL OFFICE. The Marine Resburces of the West Indies ; Ceylon P6arl Fisheries
. •. 169-178
Appendix. List of North African Locusts ........ 179-184
List of African Termites ......... 184-185
Index
186-192 I'setse-fly and its Connection with the Buffalo (Sos caffa) (Correspondence) ; 147-155
White Ants or Termites in the Sudan (Correspondence and Report)
; 155-163
Locust Plagues in the Sudan (Correspondence and Report)
. . 163-169 Appendix. List of North African Locusts ........ 179-184
List of African Termites ......... 184-185
Index
186-192 Contents. IX LIST OF FIGURES. Pig. 1. The Pigmy Mangold Beetle (Atoinaria linearis}
. Fig. 2. The Green Rose Chafer (Cetonia auratd)
. Fig. 3. False-scale, true scale and Canker-blisters on Fruit Twi
Fig. 4. Protection of Cabbage Plants from Root Maggots
Fig. 5. The Bud Moth {Hedya ocellana)
Fig. 6. Twig with larval cases of Bud Moth and Buds
Fig. 7. The Pith Moth (Zauenift a^ra). Fig. 8. Mussel Scale (^Mytilaspis pomorum)
. Fig. 9. The Potato Stem-borer (Hydriecia micacea)
Fig. 10. The Pigmy Potato Beetle {Bathyscia woUastoni)
Fig. 11. Injurious Tipididse, or Daddy-long-legs
Fig. 12. Larva and Pupa of Pissodes notatus
. Fig. 13. The Banded Pine Weevil (Pissodes notatus)
Fig. 14. Pine cone damaged by P. notatus
Fig. 15. Household Mites (Glyciphagus domesticus and G. spinipes')
Fig. 16. Coffee Berries damaged hy Arseocerus fasciculatus and Scolyti
Fig. 17. Termite protection ...... Fig. 18. A Hopper-Dozer ...... 8p.(?)
PAGES
9
14
24
35
65
66
70
75
82
85
97
116
116
117
121
137
161
166 A Classification of Animals from the point of view
of Economic Zoology. Geoup a.—Animals
captured
or
slaughtered
by man for food,
or
for
the use by him
in
other ways,
of
their
skin, bone,
fat,
or
other products. Examples.—Animals of
tlie chase
;
food-j&shes
;
whales
;
pearl-mussels. Geoup a.—Animals
captured
or
slaughtered
by man for food,
or
for
the use by him
in
other ways,
of
their
skin, bone,
fat,
or
other products. Examples.—Animals of
tlie chase
;
food-j&shes
;
whales
;
pearl-mussels. Group B.—Animals
Ired or cultivated by man for food or for the use of
their products
in
industry
or
for
their
services
as living
things. Examples.—Flocks
and
herds
;
horses
;
dogs,
poultry
gold-fish
;
bees
; silkworms and leeches. Group
C.—Animals which directly promote man's operations as a civilised
being without being killed, captured or trained by him. Examples.—Scavengers such
as vultures
;
carrion-feeding
insects
; earthworms and flower-fertilizing insects. Group
I).—Animals which concern man
as causing bodily injury, some-
times
death,
to him, and
in
other
cases
disease, often
of a
deadly character. Examples.—Lions ; wolves
; snakes
; stinging and parasitic
insects
;
disease
germ-carriers,
as
flies
and
mosquitoes
;
parasitic worms
;
parasitic protozoa. Group
E.—Animals
which
concern man
as
causing
bodily
injury
or
disease (both possibly of a deadly character) to {A), his stock
of domesticated animals
;
or {B), to his vegetable plantations
;
or
(6'),
to wild animals
in the preservation
of which he
is
interested
; or {D), wild plants
in the preservation
of which
he is interested. Examides.—Similar to those
of Group D, but also insects
and worms which destroy crops,
fruit and forest
trees, and
pests such as frugivorous birds, rabbits and voles. First Report on Economic Zoology. xii Oiioup
F.—Animals
which
concern man
as
being
destructive
to
his
worked up products of
art and
industry, such
as {A)
his
various works, buildings, larger constructions and habitations
;
(jB) furniture, books, drapery and clothing
; (C) his food and
his stores. Examples.—White
ants
;
wood-eating
larvffi
;
clothes
moths, weevils, acari and marine borers. Oiioup
F.—Animals
which
concern man
as
being
destructive
to
his
worked up products of
art and
industry, such
as {A)
his
various works, buildings, larger constructions and habitations
;
(jB) furniture, books, drapery and clothing
; (C) his food and
his stores. Examples.—White
ants
;
wood-eating
larvffi
;
clothes
moths, weevils, acari and marine borers. A Classification of Animals from the point of view
of Economic Zoology. <tROUP
Cr.—Animals which
are
known
as
" beneficials " on
account
of
their being destructive
to
or
checking
the
increase
of
the
injurious animals classed under Groups I), E and F. Examples.—Certain carnivorous and
insectivorous
birds,
reptiles and amphibia
; parasitic and predaceous insects, acari,
myriapoda, etc. Examples.—Certain carnivorous and
insectivorous
birds,
reptiles and amphibia
; parasitic and predaceous insects, acari,
myriapoda, etc. The above
is a complete
classification
of animals
in
their economic
relation to man, and proceeds from the simpler relations of primitive man
and the animals around him
to the more complex relations
of
civilised
man with his endless arts and industries and circumscribed conditions. It is, however, convenient
in the treatment of the subject, whether in
a Museum Collection
or
in
a
Handbook,
to
deal with
the
last group
(Group G), the
beneficial
animals,
in
immediate
connection
with
the
injurious animals by the destruction
of which they render
service. The
diseases of
injurious animals caused by parasitic plants such as fungi and
bacteria
ai-e naturally connected
also with
this subject of " beneficials."
But in the artificial scheme which we have decided
for
practical reasons
to accept, they are omitted, and the student is referred to the botanist and
pathologist for the treatment of these vegetable organisms. A similar treatment
of Group E, namely, those animals which injure
other animals
in the conservation of which man
is
interested, would be
convenient in some ways. But
it
is not followed here for two reasons,
firstly, because
it
is convenient rather
to
associate
this group with the
animals causing disease or death
to man, the animals
of the two groups
being in many cases identical or closely nslated, and
secondly, because tlie
zoologist has to take cognizance of
a further large and important series of
injurious animals, namely, those which destroy or injure the cultivated or
Avild plants in the life of which man is interested. Animals Captured or Slaughtered by Man for Food, or for the
use by him,
in other ways, of their
Skin, Bone, Fat, or
other Products. Animals Captured or Slaughtered by Man for Food, or for the
use by him,
in other ways, of their
Skin, Bone, Fat, or
other Products. This group
includes
those
animals
having
the
most primitive and
direct relation to man, those which he hunts and captures or kills. Perhaps
the
relation
of some
(but not
all)
of those animals which
infest or attack the body of
uncivilised man may be regarded
as equally
primitive,
that
is
to
say, the
relations
are
free from the complicating
circumstances of the civilisation of great communities of mankind. It is not desirable
in a general Museum
of Natural History to bring
together a special
series
of these animals
of the chase
or fishery. They
are best seen and
are
fully represented
in
the
general
galleries of the
Museum,
Here they may be roughly enumerated. According to locality
and circumstance, almost any animal may become the source of food or of
economic products to
this or that
race
of man. In the
list given below
those animals only
are
cited which
are regularly and
habitually pursued
by man,
either for the
piu-poses
of
procuring them
for food
or
as the
source of economic products. p
We divide the group into two sub-groups. {a) Animals pursued for food. {a) Animals pursued for food. iV) Animals pursued for their economic products. Introduction. Introduction. xiii space
at our disposal
for
the
exhibition
of
specimens
relating
to
the
subject of Economic Zoology, we find
it
safficient
to distinguish in each
group or smaller division the " British " and the " Extra-British " animals. An animal once established
as an inhabitant of Britain we shall consider
as British, whether
it
is
of foreign importation or long established as an
inhabitant of these islands. A Classification of Animals from the point of view
of Economic Zoology. It
is obvious that the subject-matter of Economic Botany could be
set forth
in a
series
of groups exactly parallel to those which we have
employed
for reviewing
the
subject-matter
of
Economic
Zoology
;
we
should merely have
to
substitute the word " plant " for " animal " in the
groups given above, and
to use
the
appropriate words
in the
place
of
" captured " and " slaughtered." A review of the contents of each
of the main Groups A to G is given
below. It
is to be noted that the animals of Group G will, as explained
above, be placed
in our Museum
series (and
in any further treatment of
the subject based on this prodromus) alongside of the
particular forms of
injurious animals to which they are hostile. It is also found convenient
in
a subject which has such definite local
interest and importance
as has that of Economic Zoology to
sub-divide
every group into a
series
of sections corresponding
to
large geographical
areas. For the purposes
of the Natural History Museum, and with the First Report on Economic Zoology. Mammals
All except the carnivora are eaten; but civilised man
is
more selective than uncivilised man, and refuses as a
rule to eat mammals not belonging to the Ruminantia,
excepting the pigs, hares, rabbits, sometimes horse, and
occasionally
his dog. The manatee
is eaten in West
Indies
; the fox bat (Pteropus
edulis) in Malay
; white
whale (Delphinaptcrns Icucas) in Greenland and Siberia. Various marsupials—the koala (Pliascolarctus cinereus)
;
wallabies and kangaroos
(Macropus)
;
rabbit-bandicoot
(Peragale lagotis), etc. Survey of Sub-group
(h) of Group A. ANIMALS PURSUED FOR THEIR ECONOMIC PRODUCTS. Protozoa
None. T'orifera
. . Sponges (grass sponge, Hippospongia equina ; wool sponges,
H. equina,
variety
gossypina ;
Zimocca
sponge,
S. zimocca ;
yellow
sponge, variety
corlosia, and
others) Crustacea
AH groups are eaten. Even
cirrhipedes {Pollicipes)
are
sold in the market at Madrid and Balanus psittacus in
South America. Arachnida
Only by uncivilised man. Hexapoda
Locusts
in
countries
where
they abound
(Greece
and
N. S. Wales)
;
larvse
of aquatic insects and midges are
compressed
into cakes in Africa (Kunga cakes)
; bees
(honey)
;
the
grubs
of palm
weevils
[RJnjncliophorus
palmariim)
are eaten by natives
of India and Burma. jNIanna produced by scale-insects (Gossyparia mannipara
in Arabia)
; ants eaten in India
; Bugong moths
eateft
in
Australia; the
Chinese
eat
the
chrysalids
of
the
silk moths. Karens
eat
cicadas
;
Kaffirs
and
East
Indians cook termites, and also eat them raw. Chilopoda
By South American Indians. Diplopoda
None. Mcllusca
Examples of
all groups are eaten either raw or cooked by
both
civilised and
micivilised
people
(oysters,
Ostrea
edulis, 0. parasitica
; clams, Mya arcnaria, species of
ilfoc/raandFen^tsandRazorshells, Ensis Americana; Ark
shells, Area and Codakia in America and West Indies). Piddocks (Pholas) are eaten in Normandy
;
snails (Helix
aspcrsa) in France; linlimus ovahis
is sold as food in
Rio Janeiro
; whelks (Buccinitm) and limpets [Patella)
in Europe
; even sea
slugs (Aplysia) are eaten in the
South Sea Islands. Tunicates
One
species,
Cynthia
microcosmus,
is
eaten raw
and
cooked by the Adriatic fishermen. Fishes
AH kinds are eaten, even in civilised countries. Fish fins
and fish maws eaten by Chinese, and isinglass obtained
from swim-bladders of sturgeons and other fish. Amphibians
Frogs only are eaten both in Europe (Rana esculenta) and
India (croaking and spangled frogs). Reptiles
Many
lizards
(Iguana
tubcrculata
in
West
Indies,
J. delicatissima in
S. First Report on Economic Zoology. First Report on Economic Zoology. xiv xiv
First Report on Economic Zoology. Crustacea
AH groups are eaten. Even
cirrhipedes {Pollicipes)
are
sold in the market at Madrid and Balanus psittacus in
South America. Arachnida
Only by uncivilised man. Hexapoda
Locusts
in
countries
where
they abound
(Greece
and
N. S. Wales)
;
larvse
of aquatic insects and midges are
compressed
into cakes in Africa (Kunga cakes)
; bees
(honey)
;
the
grubs
of palm
weevils
[RJnjncliophorus
palmariim)
are eaten by natives
of India and Burma. jNIanna produced by scale-insects (Gossyparia mannipara
in Arabia)
; ants eaten in India
; Bugong moths
eateft
in
Australia; the
Chinese
eat
the
chrysalids
of
the
silk moths. Karens
eat
cicadas
;
Kaffirs
and
East
Indians cook termites, and also eat them raw. Chilopoda
By South American Indians. Diplopoda
None. Mcllusca
Examples of
all groups are eaten either raw or cooked by
both
civilised and
micivilised
people
(oysters,
Ostrea
edulis, 0. parasitica
; clams, Mya arcnaria, species of
ilfoc/raandFen^tsandRazorshells, Ensis Americana; Ark
shells, Area and Codakia in America and West Indies). Piddocks (Pholas) are eaten in Normandy
;
snails (Helix
aspcrsa) in France; linlimus ovahis
is sold as food in
Rio Janeiro
; whelks (Buccinitm) and limpets [Patella)
in Europe
; even sea
slugs (Aplysia) are eaten in the
South Sea Islands. Tunicates
One
species,
Cynthia
microcosmus,
is
eaten raw
and
cooked by the Adriatic fishermen. Fishes
AH kinds are eaten, even in civilised countries. Fish fins
and fish maws eaten by Chinese, and isinglass obtained
from swim-bladders of sturgeons and other fish. Amphibians
Frogs only are eaten both in Europe (Rana esculenta) and
India (croaking and spangled frogs). Reptiles
Many
lizards
(Iguana
tubcrculata
in
West
Indies,
J. delicatissima in
S. America, water lizards, Varanus
dracsena, in India, and
others) are eaten
by
civilised
man
;
also
chelonians,
as
the
green
turtle,
Chelmie
midas, and the hawksbill, C. imbricata. Alligators are
eaten by Indians, and crocodiles by Siamese. _ Snakes
are eaten by Australian aborigines. Birds
All kinds eaten except birds of prey and fish-eating birds. Eggs of some wild species, as plovers and gulls. Nests
formed by swiftlets (Collocaliafucipliaga and Cfrancica)
used for soups by Chinese. Survey of Sub-group
(a) of Group A.
ANIMALS PURSUED FOR FOOD. Protozoa
None. Porifera
None. Coelentera
Sea anemones
(cul de
niulet)
are
to
be
seen
in most
French fish markets and are also eaten in Sicily, Trieste,
and Istria {Actinia viridis and others). Echinoderma
Ecliinus
(sea urchins), the ovaries
of various species
in
all parts of
the world,
especially in the West
Indies
and Adriatic Coast,
Holothurians, known
as " beche-de-mer "
or "trepang,"
are dried and cooked by the Chinese, Neapolitans and
others. Platyhelmia
Cestodes (tapeworms) are eaten by the Chinese. Nemertina
None. Nematoda
None. Chsetopoda
Palolo worms (Eunice) are eaten in the Samoan Islands
in large quantities. First Report on Economic Zoology. America, water lizards, Varanus
dracsena, in India, and
others) are eaten
by
civilised
man
;
also
chelonians,
as
the
green
turtle,
Chelmie
midas, and the hawksbill, C. imbricata. Alligators are
eaten by Indians, and crocodiles by Siamese. _ Snakes
are eaten by Australian aborigines. Birds
All kinds eaten except birds of prey and fish-eating birds. Eggs of some wild species, as plovers and gulls. Nests
formed by swiftlets (Collocaliafucipliaga and Cfrancica)
used for soups by Chinese. Mammals
All except the carnivora are eaten; but civilised man
is
more selective than uncivilised man, and refuses as a
rule to eat mammals not belonging to the Ruminantia,
excepting the pigs, hares, rabbits, sometimes horse, and
occasionally
his dog. The manatee
is eaten in West
Indies
; the fox bat (Pteropus
edulis) in Malay
; white
whale (Delphinaptcrns Icucas) in Greenland and Siberia. Various marsupials—the koala (Pliascolarctus cinereus)
;
wallabies and kangaroos
(Macropus)
;
rabbit-bandicoot
(Peragale lagotis), etc. Survey of Sub-group
(h) of Group A. ANIMALS PURSUED FOR THEIR ECONOMIC PRODUCTS. Protozoa
None. T'orifera
Sponges (grass sponge, Hippospongia equina ; wool sponges, None.
Sponges (grass sponge, Hippospongia equina ; wool sponges,
H.
equina,
variety
gossypina ;
Zimocca
sponge,
S.
zimocca ;
yellow
sponge, variety
corlosia, and
others)
are collected in many parts, as West
Indies, Florida,
Mediterranean. ANIMALS PURSUED FOR THEIR ECONOMIC PRODUCTS. hitrodtiction. xv Coelentera
Red coral (Corallium rubrum) and Isis and Mopsea. Echinoderma
Starfish
(Asterias
vidgaris)
are
collected
and
used
as
manure
(five-finger
naanure)
in some
parts
of
Great
Britain. Platyhelmia to Chsetopoda None. Crustacea
None. Arachnida
None. Hexapoda
Various insects are sought for by man for their products
lac insects
(CoccidcB)
; " cantharides " (= dried beetles,
Cantharides vesicatoria and others)
;
oil extracted from
locusts in Algeria
;
galls (Cyiiips) for dyes, tanning and
ink
;
beetles for their metallic elytra, used as ornaments
for embroidery (Chrysochtis auratus, Buprestis vittata)
;
' ground pearl
' of Bahamas produced by a scale insect,
Margarodes formicarium. '
Chilopoda
None. Diplopoda
None,
MoUusca
Many
shells
are
collected
for
ornamental
purposes
cameos (Cassis madagascarensis and others)
;
Sepia for
cuttle
bone
and
sepia
;
pearl
oysters
(Aviculidx)
;
cowries (Cyprsea moneta) are used
as money by some
uncivilised races
; the byssus
of Pinna for silk
; pearls
and
mother-of-pearl from pearl oysters {Avicula mar-
garitifera), and purple pearls from Area ; others for dyes,
as Aplysia camelus in Portugal. Tunicata (Ascidians)
None. Fish
Various sharks for
their skin (shagreen);
fish skins are
used to clarify beer, also
isinglass. Glue
is also made
from
fish skins in India. Cod
liver
oil from the cod
{Oadus tnon-htix). Fish
bile used chemically in India. Fish scales, of the bleak (Leticiscus alburnus) and dace
(L. OTtZgraris), to make artificial pearls. Fish scales of
the mahasir (Barbus
tor)
also used in manufacture of
playing cards in India. Amphibians
Frogs for their skin for ornamental book-binding. Reptiles
Many lizards and crocodiles are captured for their skins
for
ornamental purposes
;
also
turtles (hawksbill and
green turtle) for tortoise shell. Birds
The plumage of manywild birds for ornamental purposes
—grebes
(Colymbus
cristatus), skins
used
for
muffs,
trimmings, etc. ; gulls {Rissa tridactyla and others), for
hats and decoration
; Argus pheasant (Argus gigantetis)
;
peacock
feathers
in
China
;
eagle
feathers
in
N. America;
jays, cockatoos,
and
parakeets
for making
artificial
flies; Eider dncks (So77iateria wwHissima) for
eider down in Greenland, Iceland, and Norway; wild
swans for swan down
;
bile
(fel)
of the peacock used
medicinally in India. ANIMALS PURSUED FOR THEIR ECONOMIC PRODUCTS. Mammals
Most
groups
are
sought
for
for
their
(i)
furs;
(ii)
skin and hide;
(iii)
bone and horn;
(iv)
fats; .and
(v) scents,
(i) For
furs—seal,
bear, ermine, marten,
pine
marten,
sable, chinchilla, skunk, mink,
neutria, caracal,
wol-
verine,
marmot,
musquash,
genet,
squirrel,
Arctic
fox,
moles,
etc. Various
marsupials—wallabies
and
kangaroos
(^Macivpus), phalangers {Tridiosurus), and
others. (ii) For
skin
and
hide—the
wild
pig,
white
whale
(= porpoise
hide) and true
porpoises
(Pfiocfe-na
com-
munis)
;
buffalo and chamois
;
seal
for
black enamel
leather,
(iii) For
bone
and
horn—elephant
and
narwhal
for
ivory
;
'
oxen,
deer,
and
buffalo
for
horn
;
whales
(Balxna mysticettis) for whalebone. (i) For
furs—seal,
bear, ermine, marten,
pine
marten,
sable, chinchilla, skunk, mink,
neutria, caracal,
wol-
verine,
marmot,
musquash,
genet,
squirrel,
Arctic
fox,
moles,
etc. Various
marsupials—wallabies
and
kangaroos
(^Macivpus), phalangers {Tridiosurus), and
others. (ii) For
skin
and
hide—the
wild
pig,
white
whale
(= porpoise
hide) and true
porpoises
(Pfiocfe-na
com-
munis)
;
buffalo and chamois
;
seal
for
black enamel
leather, (ii) For
skin
and
hide—the
wild
pig,
white
whale
(= porpoise
hide) and true
porpoises
(Pfiocfe-na
com-
munis)
;
buffalo and chamois
;
seal
for
black enamel
leather, (iii) For
bone
and
horn—elephant
and
narwhal
for
ivory
;
'
oxen,
deer,
and
buffalo
for
horn
;
whales
(Balxna mysticettis) for whalebone. First Report on Economic Zoology First Report on Economic Zoology xvi Mammals
cont^nwiA .. (iv) For
fat, soaps and
oil—whales and porpoises, wild
pigs and bears
; spermaceti from sperm whale, (v) For
scents—musk
from
a
gland
in
the
skin
of
abdomen of male musk deer (Mosdms moschifems) in
India and used
as a stimulant medicinally, and scept
from
civets {Vivcira)
;
ambergris from sperm whaje
(Physeta macrocephalus). (v) For
scents—musk
from
a
gland
in
the
skin
of
abdomen of male musk deer (Mosdms moschifems) in
India and used
as a stimulant medicinally, and scept
from
civets {Vivcira)
;
ambergris from sperm whaje
(Physeta macrocephalus). GROUP
B. Animals Bred or Domesticated by Man
for Food,
or for the
use of their Products in Industry, or for their Services as
living things. This group
is related
to the organisation
of human
society in com-
munities possessing
fixed
dwellings, fields, stock-yards, etc. The animals
here comprised
are captured, bred and reared by man. The purposes of
this domestication by man
are
diverse, and the group may be broken up
into sub-groups
or
sections accordingly, but
they are
of
very unequal
size. The chief purposes of man's domestication of animals are
j
(a) the
provision
of
food
for
himself
;
{h) the provision
of
beasts
of
burden
{c) the
provision
of
assistance
or
companions
in
the
chase
(hounds,
ferrets, cheetah,
etc.)
;
{(V) the
provision
of guardians
for
flocks, house
and other property
;
{e) the provision
of animals which
shall amuse and
delight their owner either by brilliant plumage and
colour, song (birds),
or
1)y courage and
skill
in fighting (gamecocks,
fighting
fish)
;
(/) the
provision of hides, wool, fat, bone and other products, such as wax, honey,
silk and cochineal
;
{g) the
utilisation of the animal as a
surgical agent
(the leech). animals bred or domesticated for the provision of
assistance or as companions in the chase. Birds and Mammals alone come in this section. Birds
Hawks and falcons are used in various parts of the world
for sporting purposes. Mammalia
Various
dogs,
such
as foxhomids, deerhounds,
spaniels,
setters,
terriers
;
horses
;
elephants
; the cheetah and
ferrets. Birds
Hawks and falcons are used in various parts of the world
for sporting purposes. Mammalia
Various
dogs,
such
as foxhomids, deerhounds,
spaniels,
setters,
terriers
;
horses
;
elephants
; the cheetah and
ferrets. ANIMALS BRED OR DOMESTICATED FOR THE PROVISION
OF BEASTS OF BURDEN. Mammalia
The mammalia alone form " beasts
of burden," such as
elephants
in
India; camels
in
Africa,
etc. ; oxen
in
India and parts
of Europe
; the horse and ass, cosmo-
politan
;
the
reindeer
in
Lapland
; dogs
in
Arctic
regions and parts
of temperate Europe
;
at one time
llamas in Peru. Survey of Sub-group
(«) of Group B. ANIMALS BRED OR DOMESTICATED FOR THE PROVISION OP FOOD. Protozoa
None,
Porifera
, None. Coelentera
None. Echinoderma
None. Platyhelmia
None. Nemertina
None. Nematoda
None. Chsetopoda. . ,
None. Crustacea
Lobsters in Newfoundland
;
crayfish in France. Arachnida
... None. Hexapoda
Bees (Honey). Chilopoda
None. Dilopoda
None. MoUusca
Several kinds
of molluscs are cultivated, such as oysters
{Ostrca cdulis), cockles (Cardium cdrde), mussels (My-
tilus edulis), snails in parts of Europe (Helix pomatia). ;
Tunicata
None. Fish
Several fish are cultivated and bred for food (and sport)
salmon ((Saimo saZflr), trout (Salmo fario, S. levenensis),
rainbow
trout
[Salmo
iridcns),
land-locked
salmon
(S. sebago), whitefish (Coregonns alhus and C. ckqwi-
formis), the shad (Clupca sapidissima), carp {Cyprinits
carpio). Iiiiroduciion. xvii Amphibia
;;. Frogs are bred aud cultivated for food in parts of America
and Europe. Reptiles
Noue. Birds
Fowls, turkeys, guinea-fowls, ducks and geese,
are
culti-
vated and bred in most parts of the world. Mammals
Ruminantia, as oxen
(J3otJi(?aj), sheep (Ouirfas), and goats,
are
bred
by man
in most
parts
of the
world, even
amongst
uncivilised
tribes,
for food
; Pachyderms,
as
the pigs
; and Rodents, as
rabbits,
also
for
food. The
llama in parts of S. America, also the alpaca. Amphibia
;;. Frogs are bred aud cultivated for food in parts of America
and Europe. Reptiles
Noue. Birds
Fowls, turkeys, guinea-fowls, ducks and geese,
are
culti-
vated and bred in most parts of the world. Mammals
Ruminantia, as oxen
(J3otJi(?aj), sheep (Ouirfas), and goats,
are
bred
by man
in most
parts
of the
world, even
amongst
uncivilised
tribes,
for food
; Pachyderms,
as
the pigs
; and Rodents, as
rabbits,
also
for
food. The
llama in parts of S. America, also the alpaca. Survey of Sub-group
(7>) of Group B. Survey of Sub-group
(<^) of Group B. Hexapoda
Silk moths
Aiitherssa
mylitfa
(Tusseh
silk), Anthcrsea
pernyi. A, yavia-mai,
Attaciis cyntJiia (Ailanthus
silk),
and Bombyx viori are bred and reared in India, Cliina^
Japan and Europe
for
the
silk formed by the
larvse,
also
for the "cat-gut" made
from
the
inside
of
the
silkworms. Bees are kept by
all
civilised nations
for
the production
of honey and wax. Cochineal
insects. (Coccus
cacti)
are
cultivated
for dyes and
colours
in
Mexico, Peru, Spain, Algiers. Chilopoda
None. Diplopoda
None. Mollusca
None. Tunicata
None. Fish
None. Survey of Sub-group
(e) of Group B. Survey of Sub-group
(e) of Group B. ANIMALS BRED AND DOMESTICATED FOR THE AMUSEMENT AND"
DELIGHT OF THEIR OWNER,, EITHER BY BRILLIANT PLUMAGi;
AND COLOUR, SONG, OR BY COURAGE AND SKILL IN FIGHTING. Hexapoda
Mantis
flies and some
beetles are kept by the
Italian^
and Chinese
for fighting
;
also a grasshopper {CEticits)
;
and crickets also by Chinese. ,-[_'
Reptilia
Horned
or
Californian toads
{PJin/nosovia)
are kept ag
pets in North and Central America. The
coral snake
{Tortrix
scijtale)
is used by native women in tropical
America as an adornment round their necks. Fish
Fighting races of fish
(il/r/crq/)oJi?s ^M/5f7iflx, var.) are bred
by the Siamese. Goldfish and
others
for ornamental
purposes. Birds
Fowls (Aseels and Malays) are bred by the Malayans and
other races for fighting, and various game-fowls
(Pile
game, black-breasted reds, duckwings) in Europe for the,
same purpose. Great
varieties
of
birds are bred fo»
their brilliant plumage and colour and song—canaries,
bullfinches,
silver
pheasants
{Gennibus
nycthemcrus),
golden pheasants (Chryi>ol(yphus ficta), and others—au^
various
ornamental
ducks,
geese
and
swans,
pear,
fowls. Mammalia
Race-horses, greyhounds, and whippets,
bull dogs, pugs
and
fancy dogs. Dutch,
lop-eared, and
other fancy
rabbits. Cavies or guinea pigs. A special race of buUg
is cultivated for fighting purposes in Spain. Survey of Sub-group (/) of Group B. animals bred and domesticated for the provision of hides,
wool, pat, bone, feathers, and other products such tas
WAX, HONEY, SILK. AND MEDICAMENTS. Protozoa
None. Porifera
None. Ccelentera
None. Survey of Sub-group
(<^) of Group B. animals bred and domesticated for the provision of
guardians for flocks, house, and other property. Reptilia
The
Corn snake
{Culaber
guttatus)
is
domesticated in
N. America, and keeps off rats and mice. Rat snake in
India (Zaviciiis viucosus). Birds
The secretary bird {Gypogcranus serpcntarius) is domesti
cated by Cape farmers
for
killing
snakes,
etc.,
that
attack their animals. Mammalia
The
animals
represented in this sub-group are mainly
dogs. The common cat. Genets are domesticated in the
south of Europe for killing rats and mice. Mungooses
(HcrjKsfcs)
are
tamed
in
India,
America,
and
else-
where, and
keep
snakes,
rats, and
mice away
from
buildings
and
ships. The
hedgehog
in
Europe
for
destroying cockroaches. kviii
First Report on Economic Zoology^ Survey of Sub-group
(e) of Group B. ANIMALS BRED AND DOMESTICATED FOR THE AMUSEMENT AND"
DELIGHT OF THEIR OWNER,, EITHER BY BRILLIANT PLUMAGi;
AND COLOUR, SONG, OR BY COURAGE AND SKILL IN FIGHTING. Hexapoda
Mantis
flies and some
beetles are kept by the
Italian^
and Chinese
for fighting
;
also a grasshopper {CEticits)
;
and crickets also by Chinese. ,-[_'
Reptilia
Horned
or
Californian toads
{PJin/nosovia)
are kept ag
pets in North and Central America. The
coral snake
{Tortrix
scijtale)
is used by native women in tropical
America as an adornment round their necks. Fish
Fighting races of fish
(il/r/crq/)oJi?s ^M/5f7iflx, var.) are bred
by the Siamese. Goldfish and
others
for ornamental
purposes. Birds
Fowls (Aseels and Malays) are bred by the Malayans and
other races for fighting, and various game-fowls
(Pile
game, black-breasted reds, duckwings) in Europe for the,
same purpose. Great
varieties
of
birds are bred fo»
their brilliant plumage and colour and song—canaries,
bullfinches,
silver
pheasants
{Gennibus
nycthemcrus),
golden pheasants (Chryi>ol(yphus ficta), and others—au^
various
ornamental
ducks,
geese
and
swans,
pear,
fowls. Mammalia
Race-horses, greyhounds, and whippets,
bull dogs, pugs
and
fancy dogs. Dutch,
lop-eared, and
other fancy
rabbits. Cavies or guinea pigs. A special race of buUg
is cultivated for fighting purposes in Spain. Survey of Sub-group (/) of Group B. animals bred and domesticated for the provision of hides,
wool, pat, bone, feathers, and other products such tas
WAX, HONEY, SILK. AND MEDICAMENTS. Protozoa
None. Porifera
None. Ccelentera
None. Echinoderma
None. Platyhelmia
None. Nemertina
None. Nematoda
... None,
Chsetopoda
None. Crustacea
None. Arachnida
Spiders
have been kept
in some countries with a view
to using their webbing as silk. Survey of Sub-group (/) of Group B. Survey of Sub-group (/) of Group B. animals bred and domesticated for the provision of hides,
wool, pat, bone, feathers, and other products such tas
WAX, HONEY, SILK. AND MEDICAMENTS. Protozoa
None. Porifera
None. Ccelentera
None. Echinoderma
None. Platyhelmia
None. Nemertina
None. Nematoda
... None,
Chsetopoda
None. Crustacea
None. Arachnida
Spiders
have been kept
in some countries with a view
to using their webbing as silk. Hexapoda
Silk moths
Aiitherssa
mylitfa
(Tusseh
silk), Anthcrsea
pernyi. A, yavia-mai,
Attaciis cyntJiia (Ailanthus
silk),
and Bombyx viori are bred and reared in India, Cliina^
Japan and Europe
for
the
silk formed by the
larvse,
also
for the "cat-gut" made
from
the
inside
of
the
silkworms. Bees are kept by
all
civilised nations
for
the production
of honey and wax. Cochineal
insects. (Coccus
cacti)
are
cultivated
for dyes and
colours
in
Mexico, Peru, Spain, Algiers. Chilopoda
None. Diplopoda
None. Mollusca
None. Tunicata
None. Fish
None. animals bred and domesticated for the provision of hides,
wool, pat, bone, feathers, and other products such tas
WAX, HONEY, SILK. AND MEDICAMENTS. (vi) For other products—sugar of milk from whey of cows'
milk
;
' fel
' or purified
ox-bile
;
pepsina from mucus
membrane of the stomach of sheep, pigs, calves. Modern
medicine makes use of nearly all the glands of domesti-
cated mammalia in order to manufacture "extracts"
of a curative nature. g
(ii) For wool—sheep, such as Merinos, Lincolns, Leicesters,
Persian Lamb
;
goats,
as Angora, Kashmir or Thibet
and Sudan goats
; camels for hair which is woven into
cloth
in Persia
;
alpaca and
the
llama in Peru and
Bolivia. (iii) For
fat—pigs,
sheep,
oxen
;
prepared
suet
from
internal fat of sheep
; wool fat (^Adcps lanx)
; prepared
hog's lard. (iv) For bone—oxen, sheep, horses, and
all domesticated
animals. Horn—cattle and sheep. (iv) For bone—oxen, sheep, horses, and
all domesticated
animals. Horn—cattle and sheep. (v) For milk—cows, goats, mares. (vi) For other products—sugar of milk from whey of cows'
milk
;
' fel
' or purified
ox-bile
;
pepsina from mucus
membrane of the stomach of sheep, pigs, calves. Modern
medicine makes use of nearly all the glands of domesti-
cated mammalia in order to manufacture "extracts"
of a curative nature. (vi) For other products—sugar of milk from whey of cows'
milk
;
' fel
' or purified
ox-bile
;
pepsina from mucus
membrane of the stomach of sheep, pigs, calves. Modern
medicine makes use of nearly all the glands of domesti-
cated mammalia in order to manufacture "extracts"
of a curative nature. THE UTILISATION OF THE ANIMAL AS A SURGICAL AGENT. The chief animals coming in this sub-group are the Medicinal Leeches
SanguisHfja
mecUcinalls
and
S. officinalis. In
3Iexico
another
leech,
a
species of Hsmnentaria, is made use of. animals bred and domesticated for the provision of hides,
wool, pat, bone, feathers, and other products such tas
WAX, HONEY, SILK. AND MEDICAMENTS. Introduction. xix Introduction. xix
Amphibia
None. Reptilia
Noue. Birds
Ostriches
are farmed
for
their featliers in South Africa
and at Nice. Swans
also
to provide
feathers (swans-
down). Albumen prepared from fowls' eggs. Mammalia
The same as Sub-group (a),
i.e. cattle, sheep, goats, pigs,
and others
for
(i)
hide;
(ii) wool;
(iii)
fat;
(iv) bone
and horn
;
(v) milk
; and
(vi) other products. (i) For hide—^oxen (Bovidx), ox-hide, cow-hide, calf-skin
;
sheep-skin
for
chamois and
Morocco
leather; lamb-
skin
for
gloves. Goat-skin used
for Slorocco leather
and bottle making in the East. (ii) For wool—sheep, such as Merinos, Lincolns, Leicesters,
Persian Lamb
;
goats,
as Angora, Kashmir or Thibet
and Sudan goats
; camels for hair which is woven into
cloth
in Persia
;
alpaca and
the
llama in Peru and
Bolivia. (iii) For
fat—pigs,
sheep,
oxen
;
prepared
suet
from
internal fat of sheep
; wool fat (^Adcps lanx)
; prepared
hog's lard. (iv) For bone—oxen, sheep, horses, and
all domesticated
animals. Horn—cattle and sheep. (v) For milk—cows, goats, mares. (vi) For other products—sugar of milk from whey of cows'
milk
;
' fel
' or purified
ox-bile
;
pepsina from mucus
membrane of the stomach of sheep, pigs, calves. Modern
medicine makes use of nearly all the glands of domesti-
cated mammalia in order to manufacture "extracts"
of a curative nature. Amphibia
None. Reptilia
Noue. Birds
Ostriches
are farmed
for
their featliers in South Africa
and at Nice. Swans
also
to provide
feathers (swans-
down). Albumen prepared from fowls' eggs. Mammalia
The same as Sub-group (a),
i.e. cattle, sheep, goats, pigs,
and others
for
(i)
hide;
(ii) wool;
(iii)
fat;
(iv) bone
and horn
;
(v) milk
; and
(vi) other products. (i) For hide—^oxen (Bovidx), ox-hide, cow-hide, calf-skin
;
sheep-skin
for
chamois and
Morocco
leather; lamb-
skin
for
gloves. Goat-skin used
for Slorocco leather
and bottle making in the East. (ii) For wool—sheep, such as Merinos, Lincolns, Leicesters,
Persian Lamb
;
goats,
as Angora, Kashmir or Thibet
and Sudan goats
; camels for hair which is woven into
cloth
in Persia
;
alpaca and
the
llama in Peru and
Bolivia. (iii) For
fat—pigs,
sheep,
oxen
;
prepared
suet
from
internal fat of sheep
; wool fat (^Adcps lanx)
; prepared
hog's lard. (iv) For bone—oxen, sheep, horses, and
all domesticated
animals. Horn—cattle and sheep. (v) For milk—cows, goats, mares. First Report on Economic Zoology. First Report on Economic Zoology. XX Survey of Group
C. ^
Protozoa
.. None. Porifera
None. Ccelenteia
None. Echinoderma
None. Platyhelmia
None. Nemertina
None. Nematoda
Various eel-worms (Anguilliilidge) hasten decay in plants. Chsetopoda
The earthworms
form
surface
soil
and bury
stones on
the surface, and prepare seed beds, etc., for plants. Crustacea
None. Arachnida
None. Chilopoda
None. Diplopoda
None. Hexapoda
Many insects act as scavengers in
all parts of the world;
the
larvae
of
flies
[Calliphcyra)
feed upon and
hasten
decay of carrion
; some carrion beetles [SilphidsR)
; dung
beetles
{CopridiB
and
Dynastidx)
;
burying
beetles
(Necrophonis)
also
act
as
scavengers;
humble
bees
{Bo7nbi) fertilise clover, and many other insects act
as
fertilisers {Trigona and Vanilla, Blastopliaga and 15gs). Mollusca
None. Tunicata
None. Fish
Pish, especially carp, keep water free from insects and decay-
ing matter, and are used for such purposes in reservoirs. Amphibia
None. Reptilia
None. Birds
Vultures, by destroying the flesh of dead animals and man. Mammals
Rats feed upon
carrion, as well as sound food
; and also
jackals and semi-wild dogs. Survey of Group
C. ^
Protozoa
.. None. Porifera
None. Ccelenteia
None. Echinoderma
None. Platyhelmia
None. Nemertina
None. Nematoda
Various eel-worms (Anguilliilidge) hasten decay in plants. Chsetopoda
The earthworms
form
surface
soil
and bury
stones on
the surface, and prepare seed beds, etc., for plants. Crustacea
None. Arachnida
None. Chilopoda
None. Diplopoda
None. Hexapoda
Many insects act as scavengers in
all parts of the world;
the
larvae
of
flies
[Calliphcyra)
feed upon and
hasten
decay of carrion
; some carrion beetles [SilphidsR)
; dung
beetles
{CopridiB
and
Dynastidx)
;
burying
beetles
(Necrophonis)
also
act
as
scavengers;
humble
bees
{Bo7nbi) fertilise clover, and many other insects act
as
fertilisers {Trigona and Vanilla, Blastopliaga and 15gs). Mollusca
None. Tunicata
None. Fish
Pish, especially carp, keep water free from insects and decay-
ing matter, and are used for such purposes in reservoirs. Amphibia
None. Reptilia
None. Birds
Vultures, by destroying the flesh of dead animals and man. Mammals
Rats feed upon
carrion, as well as sound food
; and also
jackals and semi-wild dogs. GROUP
D. Animals which concern Man
as causing bodily injury, some-
times death, to him, and
in other cases disease, often of a
deadly character. This large group contains representatives
in most
of the divisions of
the
animal
kingdom. It may
conveniently be
divided
into two sub-
groups,
viz.,
(a) animals which
cause
injury, by
direct
attack,
to man,
and {h) animals which cause disease by acting as germ carriers. In the former section parasitism plays an important roh. Animals which directly promote Man's operations as a civilised
being, without being killed, captured or trained by him. Animals which directly promote Man's operations as a civilised
being, without being killed, captured or trained by him. This is a remarkable group—remarkable because
it
is
so small. The
fact
is
that
in
more
primitive
conditions
of
civilisation man
would
recognise
more
clearly
than
he
now
does
his
indebtedness
to
other
animals, as, for instance, the fisherman looks to the sea-gulls for guidance. Highly
civilised man has almost completely
separated
himself from the
ancient association with the animal world, excepting where he has seized
and
domesticated
or
more
or
less
trained
the
useful
animal. The
scavenger animals and
birds and the earthworms still act freely for man's
benefit without submitting to his yoke. 1) 2 Survey of Sub-group
(a) of Group D. ANIMALS WHICH CAUSE INJURY, BY DIRECT ATTACK, TO MAN. ANIMALS WHICH CAUSE INJURY, BY DIRECT ATTACK, TO MAN. Protozoa
Malarial
hsemamoebidse,
coccidia,
Amceba
coli,
and
Trypanosoma in the blood. Porifera
None. Coelentera
Jelly-fish, by stinging man when in sea water. Echinoderma
Spines of £c/irM«s may penetrate the skin. Platyhelmia
Numerous
tapeworms
(Ccstoda)
are
parasitic
in
the
intestines
of man
(Teenia
solium,
Tsenia
saginata),
and
in
their cystic
or
hydatid
stage
in the various
organs and
connective
tissue
(Echinococcus
hominis). Flukes (Trcmatoda)
also
occur
as
parasites
in man
(BiUiarzid Timmatohium, Distomum hepaticum). Introduction. xxi Nemertina
None,
Nematoda
Many species occur as parasites in the alimentary canal
(Ascarids and Ankylostomiivi), in the blood system and
connective tissues {Filarix), and cause disease (Elephan-
tiasis)
; the Guinea worm (Filaria mcdinensis)
; TrichinOr
spiralis. Chsetopoda
Land
leeches
[Hxmadispa
ccylonica)
attack man
in
Ceylon and India and elsewhere. Crustacea
Crabs and lobsters bite, also land-crabs (Gccarcinidas). Arachnida
A few spiders (Mygalc) have poisonous bites; mites {Sar-
coptidaa) produce
itch,
etc., and the sting of scorpions
is poisonous. Chilopoda
Centipedes (ScotojJC7icZra gigantca, S. 7)wrsita)is) in tropical
climates are poisonous, and produce painful wounds. Diplopoda
None. Hexapoda
Insects
of
the
orders
Diptera
(flies) and
Hemiptera-
Heteroptera
(bugs)
bite
in
all
parts
of
the
world. Biting Diptera include :—mosquitoes (Cidicidie), sand-
flies
{Sirmdidee),
gadflies
(Tabanidx),
stinging-flies
[Chnjsops,
Stcntioxys),
tsetse-flies
(Glossina). Diptera
also live as parasites in their adult stage (jigger-flea) and
in the larval state in man (Dermatobia, Screw-worm). Bugs of the genera Cimex and Conorhiiins. Mollusca
The
bites
of
some
are
poisonous
(Cuntisa
aulicus
in
Moluccas,
C. tcxtilis in South Sea Islands, and most
other Toxiglossa),
Tunicata
None. Fish
Sharks and various poisonous fish, the latter (i) poisonous
as food, Cliipea thrissa, C. vcncnosa ;
species of Scants,
Tetrodon,
Diodon,
^alistcs,
Ostracioii
;
the
roes
of
barbel, pike
and
burbot, and (ii) on
account
of
the
poisonous
wounds
they may
produce—weavers (Tra-
chinus), stinging rays (Trygonidx), species of Synanccia
and Thalassophryne. Amphibia
None. Reptilia
Many snakes
are
poisonous
(rattlesnake,
cobra,
fer
de
lance, blue adder, pufi adder, purple and short death
adders) and a single lizard (the Heloderma). Crocodiles
and alligators may devour man
(the
gavial,
Gavialis
gangetictis, Crocodilus vulgaris). Birds
None. Mammals
Most
of the large carnivora (lions, tigers, bears, wolves)
may cause bodily injury to man and often devour him. Any large mammals such as rhinoceros, elephant, deer,
may attack and injure him. First Report on Economic Zoology. First Report on Economic Zoology. xxii jMollusca
Some moUasca may carry germs, as oysters and cockles
carry typhoid. Tunicata
Noue. Fish
Noue. Mammalia
The imperfectly cooked
flesh
of various mammals may
carry disease to mau, as the flesh of pigs (cysts of Tssnia
suliam, and Trichina spiralis, thus producing tapeworm
and trichinosis in man)
;
the
flesh
of
oxen
(cysts
of
tapeworms, Txnia sagiaata)
;
milk
of cow and other
animals carries tuberculosis. jMollusca
Some moUasca may carry germs, as oysters and cockles
carry typhoid. Tunicata
Noue. Fish
Noue. Mammalia
The imperfectly cooked
flesh
of various mammals may
carry disease to mau, as the flesh of pigs (cysts of Tssnia
suliam, and Trichina spiralis, thus producing tapeworm
and trichinosis in man)
;
the
flesh
of
oxen
(cysts
of
tapeworms, Txnia sagiaata)
;
milk
of cow and other
animals carries tuberculosis. GROUP
E. Animals which concern Man as causing bodily injury or disease
(both possibly of a
deadly character)
to
{A)
his stock of
Domesticated Animals
; or {B) to his Vegetable Plantations
;
or
{G)
to Wild Animals; or
(D) Wild
Plants
in the pre-
servation of which he is interested. The examples coming
in
this group
are somewhat similar to those in
Group U, but in addition must be included
also worms and insects, which
destroy crops and fruit, garden produce and forest trees, and such pests as
the frugivorous
birds, rabbits and
voles. This forms the
largest group,
and
is directly connected with
tlie
cultivation and improvement
of land
by tillage and
forestry, and the animals used by man
in carrying on his
work, and kept for the improvement of the soil aud food. The sub-groups may conveniently be
divided
into smaller groups or
sections. ANIMALS WHICH CONCERN MAN
AS CAUSING BODILY INJURY OR
DISEASE (BOTH POSSIBLY OP A DEADLY NATURE) TO HIS STOCK
OF DOMESTICATED ANIMALS. This sub-group may be
divided
into
smaller
groups
or
sections
as
follows
:—Animals
which
concern man
as
causing
bodily
injury
and
disease to his stock of domesticated
animals,
i.e. (a) cattle
;
(/S) sheep and
goats
;
(y) horses, asses and mules
;
(8) the pig
;
(c) elephant and camel
iX) guinea pig
;
(ji) dogs and cats
;
{p)
rabbits, and (i) poultry. animals which cause disease by acting as germ carriers. Protozoa to Crustacea
None. Arachnida
Probably
ticks
(Ixodidx) may
distribute
disease
from
animals to man. Chilopoda and Diplopoda
None. Hexapoda
All
piercing-mouthed
insects may
either
carry
disease
direct from man to man or animals to man
gadflies
(Tabanidx), Stomoxys and
tsetse-fly
{Muscidse),
sand-
flies (Simulidse)
; bugs [Cimex a.nA. Conorhinus)
; or they
may act as intermediate hosts of parasites (mosquitoes
and malaria and filariasis). Germs may also be carried
to food and drink by dung-feeding
flies {Musca,
Calli-
phora,
Scatqphaga)
from
latrines
and
foul
matter
(typhoid). Arachnida
Probably
ticks
(Ixodidx) may
distribute
disease
from
animals to man. Chilopoda and Diplopoda
None. Hexapoda
All
piercing-mouthed
insects may
either
carry
disease
direct from man to man or animals to man
gadflies
(Tabanidx), Stomoxys and
tsetse-fly
{Muscidse),
sand-
flies (Simulidse)
; bugs [Cimex a.nA. Conorhinus)
; or they
may act as intermediate hosts of parasites (mosquitoes
and malaria and filariasis). Germs may also be carried
to food and drink by dung-feeding
flies {Musca,
Calli-
phora,
Scatqphaga)
from
latrines
and
foul
matter
(typhoid). Arachnida
Probably
ticks
(Ixodidx) may
distribute
disease
from
animals to man. Chilopoda and Diplopoda
None. Hexapoda
All
piercing-mouthed
insects may
either
carry
disease
direct from man to man or animals to man
gadflies
(Tabanidx), Stomoxys and
tsetse-fly
{Muscidse),
sand-
flies (Simulidse)
; bugs [Cimex a.nA. Conorhinus)
; or they
may act as intermediate hosts of parasites (mosquitoes
and malaria and filariasis). Germs may also be carried
to food and drink by dung-feeding
flies {Musca,
Calli-
phora,
Scatqphaga)
from
latrines
and
foul
matter
(typhoid). ANIMALS CAUSING BODILY INJURY AND DISEASE TO CATTLE. Protozoa
None. Platyhelmia
TargQwovxn.?, [Moniczia lylanissima) and cysts (Cysticercus
bovis, C. tenuicoUis) produce disease
;
also flukes {Disto-
inum liepaticum, D. rtiagna). Nematoda.. Numerous
thread
and
round
worms
{Ascaris,
Tricho-
ccplialus, etc.). Chsetopoda
None. Introduction. Introduction. xxi i i Introduction. xxi i i
-Arachnida
Ixodidx or ticks (Ixodes) and mange mites (Sarcc^tidsR). Hexapoda
Biting
flies (Tabamis, Hmmatopota,
Siviulium), warble
flies [Hypoderma lineata and H. bovis). Amphibia
None. Beptilia
Poisonous
snakes
attack
cattle—puff
adder;
crocodiles
{Crocodilus vulgaris) in African rivers. Birds
None. Mammals
The larger carnivora attack cattle, especially leopards. II. As germ carriers. Arachnida
Ticks
(Ixodidx)
distribute
various
bovine
diseases,
as
Texas fever. Hexapoda
Probably biting
flies, such as TabanidiB, Stomoxys, carry
disease germs (Anthrax)
;
tsetse-fly and Nagana. -Mollusca
Certain
species
of
water
snails
(Limns&idae)
carry
the
germs of flukes. -Arachnida
Ixodidx or ticks (Ixodes) and mange mites (Sarcc^tidsR). Hexapoda
Biting
flies (Tabamis, Hmmatopota,
Siviulium), warble
flies [Hypoderma lineata and H. bovis). Amphibia
None. Beptilia
Poisonous
snakes
attack
cattle—puff
adder;
crocodiles
{Crocodilus vulgaris) in African rivers. Birds
None. Mammals
The larger carnivora attack cattle, especially leopards. II. As germ carriers. Arachnida
Ticks
(Ixodidx)
distribute
various
bovine
diseases,
as
Texas fever. Hexapoda
Probably biting
flies, such as TabanidiB, Stomoxys, carry
disease germs (Anthrax)
;
tsetse-fly and Nagana. -Mollusca
Certain
species
of
water
snails
(Limns&idae)
carry
the
germs of flukes. ANIMALS CAUSING BODILY INJURY AND DISEASE TO SHEEP AND GOATS. I. By direct injury. Protozoa
Protozoa
are
said
to
cause
a
disease
in
the
feet
of
sheep in Australia. Platyhelmia
Numerous cestodes (Moniezia expansa, planissirna, alba)
in intestines and cysts in the body (Cysticercus temci-
collis), and
in the brain (Ccenurus
ccrebralis), causing
" sturdy."
The
liver
fluke
(Distomum
Jiepaticum)
producing " rot."
Nemertina
None. Nematoda
Numerous filaria in
all sheep (Ivmg worms, Eustrongylus
filaria and husk). Strongylus contortus in intestines. Arachnida
Sarcoptidx, producing scab (Psoroptes communis v. ovis)
;
Ixodidx or ticks. Chilopoda and Diplopoda
None. Hexapoda
Keds (Melophagus ovinus), nasal
fly
(CEstrus
ovis), and
sheep maggots (Lucilia), lice (Mallophaga) in sheep and
goats. Beptilia
Many poisonous snakes (puff adder, cobra). Amphibia
None. Birds
Some
birds
of
prey
(eagles);
the
kaka
parrot
(Nestor
meridio7ialis) attacks sheep in New Zealand. Mammals
Large carnivora abroad and dogs generally. II. As germ carriers. Arachnida
Ixodidx, and louping ill (Ixodes reduvius), and heartwater
{Amblyom7na). Hexapoda
Biting flies (Tabanus, Chrysops) may carry anthrax. Mollusca
Some water
snails (Limnxidx)
carry the germs
of the
fluke. Survey of Section
(y) op Sub-geoup A. ANIMALS CAUSING BODILY INJURY AND DISEASE TO HORSES,
ASSES, AND MULES. I. By direct injury. Protozoa to Echinoderma None. Platyhelmia
A few occur in horses (Tgsnia plicata, and per-foliata),
Nemertina
None. Nematoda
Many in all parts of the body; armed strongyles (ScZero-
stomum
tctracanthum and equinum)
;
thread worms
Filaria)
; maw worms (Oxyuris curvula and mastigoides). ANIMALS CAUSING BODILY INJURY AND DISEASE TO HORSES,
ASSES, AND MULES. I. By direct injury. Protozoa to Echinoderma None. Platyhelmia
A few occur in horses (Tgsnia plicata, and per-foliata),
Nemertina
None. Nematoda
Many in all parts of the body; armed strongyles (ScZero-
stomum
tctracanthum and equinum)
;
thread worms
Filaria)
; maw worms (Oxyuris curvula and mastigoides). Protozoa to Echinoderma None. Platyhelmia
A few occur in horses (Tgsnia plicata, and per-foliata),
Nemertina
None. Nematoda
Many in all parts of the body; armed strongyles (ScZero-
stomum
tctracanthum and equinum)
;
thread worms
Filaria)
; maw worms (Oxyuris curvula and mastigoides). ANIMALS CAUSING BODILY INJURY AND DISEASE
IN
PIGS. I. By direct injury. Protozoa
Balantidium coli causes ill-health. Porifera to Echinoderma. None,
Platyhelmia
Cysticerciis
cellulosse, cyst
of human tapeworm (measles
in pigs)
; no sexual tapeworm. Echiiwcoccus frequent
in the liver. Nemertina
None. Nematoda
Ascaris suilla in intestines and others; Ecliinorhynchus
gigas
;
Trichina spiralis ;
Strongylidse (S. paradoxus) in
bronchi
;
Steplienurus dentatus. Chsetopoda
None. Arachnida
Itch mites
(Sarcoptes
sca6ei) transmittable to man, and
De7nodex. Diplopoda and Chilopoda
None. Hexapoda
Stomoxys
(biting
or
stinging
fly)
;
MallopJmga
and
Hxmatopinus (lice). Amphibia
None. Reptilia
Poisonous snakes rarely attack the pig. Birds
None. Mammals
Larger carnivora,
II. As germ carriers. Protozoa to Birds
None known. Mammalia
Rats carry trichinosi s
( Trichina spiralis)
Survey of Section
(e) of Sub-group A. ANIMALS CAUSING BODILY INJURY AND DISEASE
IN ELEPHANTS
AND CAMELS. I, By direct injury. Protozoa
None. Porifera and Echinoderma None. Platyhelmia
Amphistomes cause sickness in Indian elephants; Trema^
todes in lungs
of camel
; also
Gysticercus tenuicollis in
camels; Echinococcus in liver. Nemertina
Nona. Nematoda
Strongylus filaria in the bronchi of camels. Arachnida
Pentastomes {Linguatula) in camel
; Ixodes cam^linus and
Galeodes aratwides, a great camel pest, and Sarcoptidm,
Ohilopoda and Diplopoda
None. First Report on Economic Zoology.- First Report on Economic Zoology.- XX iv Aiachnida
Acari
produce mange
and
sores
{Sarcoptcs,
Psoraptes]'
Sy^yibiotcs). Hexapoda
Biting &ies (Tabanus, Hssinatojyota)
; warble flies (Gosiro--
jyJnlus) (" bots ")
;
forest flies [Hippohoscidx). Mollusca
None. Fish
Electric
eel
{Electrophoiits
clectricus)
attacks- horses at
water in Brazil. Amphibia
None. Reptilia
Many poisonous snakes, especially the pufE
addfer, fer de
lance and others, and crocodiles. Birds
None. Mammalia
A few large carnivora, as leopards
; vampire-bats'( Va/>npyru&
spectrum). II. As germ carriers. Survey of Section (Q op Sub-gboup A.
AlflMALS CAUSING BODILY INJURY AND DISEASE
IN GUINEA
PIGS. Protozoa
Coccidiaand Infusoria {Monocercomonas cavias) live in the
intestines. Porifera to Echinoderma. None. Platyhelmia
Trematodes or flukes (Distomum cavise). Nemertina
None. Nematoda
None (?). Arachnida
Pentastomes (Linguatala) occur in the intestines. Hexapoda
None kno\vii except fleas (Pulicidas). Reptilia
Various poisonous snakes. Amphibia
None. Birds
Most rapacious birds. Mammals
Most large carnivora. ANIMALS CAUSING BODILY INJURY AND DISEASE
IN ELEPHANTS
AND CAMELS. Protozoa
None. Porifera and Echinoderma None. Platyhelmia
Amphistomes cause sickness in Indian elephants; Trema^
todes in lungs
of camel
; also
Gysticercus tenuicollis in
camels; Echinococcus in liver. Nemertina
Nona. Nematoda
Strongylus filaria in the bronchi of camels. Arachnida
Pentastomes {Linguatula) in camel
; Ixodes cam^linus and
Galeodes aratwides, a great camel pest, and Sarcoptidm,
Ohilopoda and Diplopoda
None. Introduction. xxv Hexapoda
CEstrus
cameli, common
in
camels,
and
biting
flies
(Tabanus, Chrysops). Reptilia
None
(?). Amphibia
None (?). Birds
None. Mammals
None. II. As germ carriers. None known. Hexapoda
CEstrus
cameli, common
in
camels,
and
biting
flies
(Tabanus, Chrysops). Reptilia
None
(?). Amphibia
None (?). Birds
None. Mammals
None. II. As germ carriers. None known. Survey of Section
(S) of Sub-group A.
ANIMALS CAUSING BODILY INJURY AND DISEASE TO RABBITS. I. By direct injury. Protozoa
Coccidia produce disease in the liver (liver-rot). Porifera to Echinoderma. None. -Platyhelmia
Numerous tapeworms in the intestines and cysts in the
organs
and
tissues
(Txnia
i^cctinata,
Cysticcrcus
jnsciformis)
Nemertina
None. Nematoda
Strongylidx
often
cause
great
mortality
(Strongylus
strigosiis, Oxyuris ainbigua). Chsetopoda and Crustacea
None. Arachnida
Psoroptcs produce scab and itch. Hexapoda
Fleas {PuLcx gonioccplialus). Reptilia
Various snakes. Birds
Rapacious birds (hawks, falcons and crows). Mammals
The fox, dogs, cats, weasels, stoats, etc. II. As germ carriers. None known. II. As germ carriers. None known. ANIMALS CAUSING BODILY INJURY AND DISEASE TO DOGS AND
CATS. I. By direct injury. Protozoa
Protozoal forms in the blood cause malignant jaundice in
dogs
in
Africa. Coccidium pcrforans in intestines [of
dog. Porifera to Echinoderma. None. Platyhelmia
Numerous Cestodes infest dogs (Tasnia cxmirus, T. mar-
ginata, T. echinococcus). T. crassicollis in cats. Nemertina
None. Nematoda
Various species in both
cat and dog (Ascaris mystax in
cats; Filaria inimitis in dogs). Arachnida
Ticks (Ixodidm)
; Mange Insects (Psoroptes, Symbiotes, and
Dcmodecidx), and Pentastomes. Chilopoda and Diplopoda
None. Hexapoda
Fleas
(Pulicidas) and
lice {Mallophaga) on hoih dog a,ndi
oat. Reptilia
Poisonous snakes, and especially the alligator {AlUgator
Mississippiensis) . Amphibia
None. Birds
None. Mammals
Other large carnivora. II. As germ carriers. Protozoa to Crustacea
None. Arachnida
A Tick {Ixodes sp.) carries germs of malignant jaundice. Hexapoda
Culicidas carry the embryos
of
Filaria
immitis. Lice
(Trichodectes) , the cysts of Tasnia cdnina. Chilopoda to Birds
None. Mammals
Sheep, rabbits, hares and mice contain hydatids of sorne
canine
and
cat
tapeworms
{Ca!nurus
cerebralis
in
sheep, Cysticercus piscifo7-mis in hares and rabbits). -"XXV i
First Report on Economic Zoology. Introduction. Introduction. xxvii xxvii Survey of Suu-oroup B of Group E.
animals which concern man as causing injury and
disease to his vegetable plantations. This group deals with animals which cause
loss to farmers, gardeners
and
foresters. It may then
be
divided
into three main
sections
;
(i.)
animals
injurious
to
agriculture
;
(ii.)
to
horticulture
;
and
(iii.)
to
forestry. Each
of these
sections may be
again conveniently
divided up
into
smaller groups. Survey of Section
I. of Sub-group B. ANIMALS IXJURIOU.S TO AGRICULTURE. This section may be divided up into several headings under the names
of the particular crops grown by man. It is not possible here to enter at
length into this sub-group, owing to
its extensive nature. The following
divisions
of
this
section may
be
employed
:—(a) animals
injurious to
cereal crops
;
(/?)
to
pulse
;
(y)
to root crops
;
(8)
to forage crops and
grass
;
(c) to fruit and fruit trees
;
(^) to hops
;
(iy) to tea
;
{&) to coffee
;
(t) to sugar. ANIMALS CAUSING BODILY HARM AND DISEASE TO POULTRY. Protozoa
Cause
false
coccidiosis
of
liver
{Amceba
meleagris),
diphtheritic roup, epithelioma contagiosum. Platyhelmia
Tape worms often
occur
in
all
poultry {Davainea pro-
glottina, Drcpanidoteenia infundihuliformis and others). Nemertina
None. Nematoda
Thread worms (Heicrakis) live in the intestines of fowls;
the gape worm [Syngaimis trachealis, gapes). Chsetopoda
None. Arachnida
Sarcoptidx {Ae^lnvamg scabies, Sarcoptes
i^uis), scaly leg
{Sarcoptes miitans)
; ticks (Argas)
; mites (Dermanyssus
avium) on skin and feathers. Chilopoda and Diplopoda
None. Hexapoda
Lice
(Mallophaga),
fleas
(Pulicidx)
and
certain
flies
(Ornitliomyia)
and
sand
flies
(Sinmliiim)
cause
annoyance. MoUusca
None. Reptilia
Poisonous snakes attack poultry (chicken snake, Coluber
quadrivittatus in N. America and others). Birds
Hawks
(sparrov/-hawk)
and
crows
take
the
chicks
;
eagles, kites
; peregrine falcon. Mammals
Many mammals prey on poultry and
eggs
(fox,
polecat,
weasel, rat, Indian civet, leopard cat). II. As germ carriers. Chsetopoda
Earthvrorms
carry
the
ova and embryos
of
the
gape
worm and are eaten by fowls. ANIMALS INJURIOUS TO CEREAL CROPS.
The animal pests of corn crops are very numerous. Protozoa
None. Platyhelmia
None. Nemertina
None. Nematoda
Eelworms (AiiguilliUidie) cause disease,
as tulip root in
oats. Chsetopoda
Some Enchytrams apparently cause disease to plants. Crustacea
Land Isopods (woodlice) sometimes Ao h&xm. {Armadilli-
dium, Oniscus). Arachnida
None(?). Hexapoda
Numerous
insects
eat
leaf
(larvae
of
Nochix),
stalks
(Hessian
fly,
Cccidomyia
destructor, maize
aphis,
A. viaiis, Ccphiis pyginanis, Leucanium, etc.),
roots
(wire-
worm, Elatcridas), and seed and blossom (wheat midge
Diplosis tritici). Chilopoda
None. Diplopoda
Millepedes {Julidee) attack the roots of corn. Mollusca
Many snails (iJeZfcwZ«) and slugs
(Liwftcirfie) devour the
leaves. Reptilia and Amphibia
. . . None. Birds
Destroy the seed and pull up young plants (rooks, starlings,
wood-pigeons,
cranes
{Gruidie),
wild
geese
{Anser
anscr in Europe, Plectroptcrus gcnnbcnsis in Transvaal). Mammals
Ruminantia, especially deer,
often do harm to standing
corn
;
rodents, as voles and mice. Survey of Sub-section
(/3) of Section I. ANIMALS INJURIOUS TO PULSE. Protozoa to Nemertina
. . . None. Nematoda
Eel-worms (Tylcnchtcs). Crustacea
...;
Land Isopods (woodlice) attack young plants
{Porcellio,
Oniscus). First Repoi^t on Economic Zoology. Aiachnida
None (?). Hexapoda
Many insects attack leafage (pea weevils,
Sitones), stem
(Ai)hides),
roots
(wire-worm
or
Elatcr
larvae),
seed
(pea moth, Grapliolitha pisana),
Chilopoda and Diplopoda
None. Mollusca
'. Snails (Helix) and slugs (Limax). Reptilia and Amphibia
... None. Birds
Many of the passerine birds take the seed in the ground
;
also pigeons, rooks and jackdaws. Mammals
Mice take seed in the ground. Survey of Sub-section (y) of Section
I. ANIMALS INJURIOUS TO ROOT CROPS. Protozoa to Nemertina
. . . None. Nematoda
None (?). Crustacea
Woodlice (Onisciis, Ar77mdillidium.). Arachnida
None (?). Chilopoda
None. Diplopoda
Many Jididep, damage roots. Hexapoda
Large numbers
of
insects attack root crops (flea beetles
(Halticidie)
;
surface
larvae
(Noctitge);
diamond-back
moth
(Plntella
maculipcnnis)
;
root
weevils
(Ceuto-
rhynchus) and Aphides). Mollusca
Slugs and snails (Liwacid* and BieZicidas). Beptilia and Amphibia
... None. Birds
Many
birds
eat the
seedlings
(linnets, sparrows, green-
finches, larks). Mammalia
Rabbits, hares and deer. Survey of Sub-section (8) of Section I. ANIMALS INJURIOUS TO FORAGE CROPS AND GRASS. Protozoa to Nemertina
... None. Nematoda
Tylenchus devastatrix causes clover sickness. Arachnida
None. Hexapoda
Large numbers
attack roots (chafer
larvae,
Melolontlia,
,«. Cetonia, leather jackets or
larvae
of
Tipulidie), leaves
(weevils
Apions),&ni seeds (thrips, midges
Diplosis). Chilopoda and Diplopoda
None. Mollusca
Slugs and snails attack young forage crops. Reptilia
None. Amphibia
None. Introduction. Introduction. xxix Amphibians and Reptilia
None. Birds
Many frugivorous
birds,
hornbills
(Bitcerotidse),
trogons
(Trogonoideie),
cockatoos
and
parrots
{Plissoloijhus
moliiccensis),
depredate
whole
fields
of
fruit,
in
Moluccas
;
ring parrot (Palieoniis torqjuitjis), in Africa
and
India, grey parrot (Psittacus
erithaciis) in Africa,
do much harm in
fruit plantations
; others do so now
and
then;
starlings
{Stiirnus),
thrushes
{Turdid,-e),
toucans {Rhamphastidse)
; other birds damage twigs, etc. (plant cutters, Phyfotomidx). Mammalia
Rabbits
and
hares damage young trees
by
biting
the
bark
:
squirrels take nuts and soft fruit, and some fruit
bats
{PtcrapuH
imJioccphalns,
P. Kcrandrcnii),
and
others do mucli harm abroad. ANIMALS INJURIOUS TO CEREAL CROPS.
The animal pests of corn crops are very numerous. Birds
The seed of grass, clover, etc., is eaten by sparrows and
finches. Mammalia
Moles cause annoyance in pastures by throwing up hills
gophers in America
;
voles and rabbits. Survey of Sub-section (c) of Section
I. ANIMALS INJURIOUS TO FRUIT AND FRUIT TREES,
Protozoa to Chaetopoda
... None. Crustacea
Woodlice damage soft fruits (Oniscits, Asellus). Arachnida;
Various
mites
(red
spider,
Tctranychtis
and
Bryobia)
damage the leafage. Hexapoda
Hosts of insects attack fruit—codling moth, woolly aphis,
San Jos6 scale, pear midge, mussel scale, bark
beetles,
plum cuculio, fruit flies {Ceratitis), phylloxera. Leafage,
•
fruit, stem and roots are all subject to insect ravages in
all parts of the world. Chilopoda
None. Diplopoda
Some millepedes (Julidx) damage soft fruits. Mollusca
Slugs and snails damage soft fruits on the ground. ANIMALS INJURIOUS TO HOPS. Protozoa to Nemertina
. . None. Nematoda
Eel-worms (Hctcrodcra), produce disease. Crustacea
Woodlice {Annadilliditiin &ndi Ascellus),
Arachnida
Red si^iders (Tetranychiis tclarius). Chilopoda
None. Diplopoda
Millepedes (Jididse) damage the roots.'
Hexapoda
Many
insects
attack
root,
leaves and stem
Aphis, fioa
beetle (Haltica conciniia), wire-worm {Agriotcs liiieatus),
hop-dogs
{D. pudibiiiula),
shy-bugs
(Calocoris
fulco-
maculatiis). MoUusca
Occasionally snails (ifcZix). Heptilia and Ampnibia
... None. Birds
Some birds, as linnets, damage the cones. Mammals
Rabbits and hares attack the bine. Survey of Sub-section
(r?) of Section I. ANIMALS INJURIOUS TO TEA. Protozoa to Chaetopoda
. . . None. Crustacea
Land crabs in India. Arachnida
Red tea mite (Tetranychus
hiaculatus)
\
fire-legged
tea
mite {Typhlodromus carinatus)
; yellow tea mite {Acarus
transluceus)
; pink mite (Phytoptus tlwx). Hexapoda
"RqA. horer {Zcuzera coffex); black grub
{Agrotis siiffnsa)
faggot worm {Eumcta carmcri)
;
tea
scale
[Aspidiotns
theas)
;
tea
aphis
{Ccylonia
tlieascola)
;
white
ants
(Termes
taprobanes)
; tea mosquito or tea bug (Hclio-
peltis theivora), and otliers. Chilopoda and Diplopoda
None. MoUusca
Snails and slugs do much harm (Hcliocarion salius a,nd. others). Reptilia and Amphilia
... None. Birds
None. Mammals
Wild elephants do much harm in tea plantations;
also
stray cattle, hares and laud rats. Survey of Sub-section (6) of Section I. ANIMALS INJURIOUS TO COFFEE. P'rotozoa co Crustacea
None. Arachnida
None
recorded,
but
probably
several
occur
on
coffee
leaves. ANIMALS INJCRIOUS TO SUGAR-CANES. Protozoa to Nemertina
... None. Nematoda
Several
damage
sugar
cane
(Hctcro<iera
saccTeari,
Tij-
lenchiis saccJiari, and others). Arachnida
Numerous mites damage leafage and stems (Histiostoma,
rostroserrat lis, Tarsonijmus Bancroftii). Hexapoda
Sugar-cane
borers [Diatrxa soccharalis)
', tropical sugar-
cane borer
[Cltilo
saccharalis)
; pin
borers
{Xylebarus
jncens
and
X. pcrforans
;
Sandwich
Island
borers
{Sphcnopliorus
obscurus)
;
white
grub
of
Queensland
{Ijcpidiota
sqnamulata)
;
sugar
scale
{Aspidiotus sac-
cJiari). Chilopoda and Diplopoda
None. Mollusca
None. Beptilia and Amphibia
... None. Mammalia
Rats and stray domestic stock. Protozoa to Nemertina
... None. Nematoda
Several
damage
sugar
cane
(Hctcro<iera
saccTeari,
Tij-
lenchiis saccJiari, and others). Arachnida
Numerous mites damage leafage and stems (Histiostoma,
rostroserrat lis, Tarsonijmus Bancroftii). Hexapoda
Sugar-cane
borers [Diatrxa soccharalis)
', tropical sugar-
cane borer
[Cltilo
saccharalis)
; pin
borers
{Xylebarus
jncens
and
X. pcrforans
;
Sandwich
Island
borers
{Sphcnopliorus
obscurus)
;
white
grub
of
Queensland
{Ijcpidiota
sqnamulata)
;
sugar
scale
{Aspidiotus sac-
cJiari). Chilopoda and Diplopoda
None. Mollusca
None. Beptilia and Amphibia
... None. Mammalia
Rats and stray domestic stock. ANIMALS INJURIOUS TO HORTICULTURE. The number and variety of plants
cultivated
in the garden
is so great
and so varied that it
is not possible to sub-divide them in detail. For our
purpose we may, however, divide them into two sub-sections in connection
with their animal enemies
: (a) The animals injurious to culinary plants. (a) The animals injurious to culinary plants. {(3) The animals injurious to ornamental plants, {(3) The animals injurious to ornamental plants, ANIMALS INJURIOUS TO TEA. Protozoa to Chaetopoda
. . . None. Crustacea
Land crabs in India. Arachnida
Red tea mite (Tetranychus
hiaculatus)
\
fire-legged
tea
mite {Typhlodromus carinatus)
; yellow tea mite {Acarus
transluceus)
; pink mite (Phytoptus tlwx). Hexapoda
"RqA. horer {Zcuzera coffex); black grub
{Agrotis siiffnsa)
faggot worm {Eumcta carmcri)
;
tea
scale
[Aspidiotns
theas)
;
tea
aphis
{Ccylonia
tlieascola)
;
white
ants
(Termes
taprobanes)
; tea mosquito or tea bug (Hclio-
peltis theivora), and otliers. Chilopoda and Diplopoda
None. MoUusca
Snails and slugs do much harm (Hcliocarion salius a,nd. others). Reptilia and Amphilia
... None. Birds
None. Mammals
Wild elephants do much harm in tea plantations;
also
stray cattle, hares and laud rats. Survey of Sub-section (6) of Section I. ANIMALS INJURIOUS TO COFFEE. P'rotozoa co Crustacea
None. Arachnida
None
recorded,
but
probably
several
occur
on
coffee
leaves. First Report on Economic Zoology, First Report on Economic Zoology, First Report on Economic Zoology, XXX Hexapoda
Coffee scales {Lccaninm caffex,
Aspidiotus
articnlatus)-;
mealy
bug
{Dactnlobius
destructor)
;
coffee
moth
(Hcliotlds armigcra)
;
coffee miner
(Gracillaria
coffei-
foliella)
;
also Onjzia
Ceylanica,
Zciczera
coffcm, and
others. Chilopoda and Diplopoda
None. MoUusca
Probably snails and slugs. Reptilia and Amphibia
... None. Birds
None. Mammals
The same as section
(t;). AXIMALS IXJURIOUS TO FORESTRY, The
animal
pests
of
forestry may he
best
treated
in
detail under
smaller sections dealing with allied oroups of trees
;
i.e., animals injurious
to (A)
Pine.s
;
(P)) Oak
;
(C)
"Willows, etc., but
for our purpose here
nO'
division need be made. There arc no enemies amongst the lower groups
of invertebrates until
we come to the Arachnida
i
A few attack the leaves of trees (Tctranychus). Hexapoda
Most orders occur on forest
trees—wood
borers,
as wood
wasps
{Sirrx'), goat moth {Cossus)
;
leaf eaters,
gypsy
moth (Portln'siii), tent caterpillars (Ciissiocaw^w)
; bark
beetles [Tomicus, Fissodcs). Chilopoda and Dilopoda... None
Mollusca
None. Reptilia and Amphibia
... None. Birds
Woodpeckers and others damage the trunks, and others
(crossbifk, Loxia') take cones and seeds. Capercaillzie
and 'Other grouse damage buds and young growths. Mammals
Many mammals do harm by barking trees (deer, rabbits
mice, volea, hares). Arachnida
i
A few attack the leaves of trees (Tctranychus). Hexapoda
Most orders occur on forest
trees—wood
borers,
as wood
wasps
{Sirrx'), goat moth {Cossus)
;
leaf eaters,
gypsy
moth (Portln'siii), tent caterpillars (Ciissiocaw^w)
; bark
beetles [Tomicus, Fissodcs). Arachnida
i
A few attack the leaves of trees (Tctranychus). Hexapoda
Most orders occur on forest
trees—wood
borers,
as wood
wasps
{Sirrx'), goat moth {Cossus)
;
leaf eaters,
gypsy
moth (Portln'siii), tent caterpillars (Ciissiocaw^w)
; bark
beetles [Tomicus, Fissodcs). y
g g
Mammals
Many mammals do harm by barking trees (deer, rabbits
mice, volea, hares). ANIMAIiS INJURIOUS TO CULINARY PLANTS. Protozoa to Nemertina
. . . None. Nematoda
Various
eel-worms
(Tijlcnclms,
Hetcrodcra). (Tomato
root disease). Chsetopoda
Encliytrseus and a few others damage roots. Crustacea
Land isopods (Oniscns, etc.) attack roots and seedlings. Arachnida
Red spiders and various acari damage leaves. Hexapoda
Most
groups
of
insects
attack
vegetables—onion
fly
[Fhorhia ccpetoruni), cut-worms (Nocttise), thrips, aphis,
leather jackets {TipHlidm). Chilopoda
None. Diplopoda
Millepedes attack various roots (Julus, Polydcsvms, etc.),
Mollusca
Snails and slugs attack delicate leaves. Amphibians
None. Reptiles
None. Birds
Several finches, the sparrow, and other small birds, as long-
tailed tits, take seeds, buds, and fruit. Mammals
Mice, rats, voIgs, skunks, and others do damage in gardens. Protozoa to Nemertina
. . . None. Nematoda
Various
eel-worms
(Tijlcnclms,
Hetcrodcra). (Tomato
root disease). Chsetopoda
Encliytrseus and a few others damage roots. Crustacea
Land isopods (Oniscns, etc.) attack roots and seedlings. Arachnida
Red spiders and various acari damage leaves. Hexapoda
Most
groups
of
insects
attack
vegetables—onion
fly
[Fhorhia ccpetoruni), cut-worms (Nocttise), thrips, aphis,
leather jackets {TipHlidm). Chilopoda
None. Diplopoda
Millepedes attack various roots (Julus, Polydcsvms, etc.),
Mollusca
Snails and slugs attack delicate leaves. Amphibians
None. Reptiles
None. Birds
Several finches, the sparrow, and other small birds, as long-
tailed tits, take seeds, buds, and fruit. Mammals
Mice, rats, voIgs, skunks, and others do damage in gardens. Introdiictwn. xxxi ANUIALS INJURIOUS TO ORNAMENTAL PLANTS. Protozoa to Nematoda
... None. Chsetopoda
Earthwonus often do harm to potted plants. Crustacea
Land isopods are destructive, especially under glass. Arachuida
Many acari cause harm {Tcfrawfchns, Bryobia). \
Hexapoda
Most orders of
insects are injurious (carnation maggot,
narcissus flj' (Mcrodon cqncsfris), scales (Chionfcsjns rosee,
Lccanium
olex),
thrips
(Thripidae),
mealy
bug (Dac-
fi/lobius),
rose
sa^\'flies
{Hijlotoma
rosse, Blcnnocampa
2)Hsilla). Chilopoda
None. Diplopoda
Millepedes
[Jnlidiv^
often
attack
ornamental
plants,
especially bulbs. Mollusca
Snails and slugs {Hclicidai and Liiiiacidx). Amphibia and Reptilia
... None. Birds
Small
birds
take
plant
seeds
(finches,
sparrows),
also
damage the blossoms. Mammals
Moles, rabbits, mice, voles, and
rats all do harm amongst
ornamental plants. GKOUP
F. Animals which
concern Man
as
being
destructive
to
his
worked-up Products of Art and Industry, such as [A) his
various Works, Buildings and
larger Constructions and
Habitations
;
(B)
his Furniture and
Books, Drapery and
Clothing
;
{C) Food and Clothes. Animals which
concern Man
as
being
destructive
to
his
worked-up Products of Art and Industry, such as [A) his
various Works, Buildings and
larger Constructions and
Habitations
;
(B)
his Furniture and
Books, Drapery and
Clothing
;
{C) Food and Clothes. The numerous animal
pests coming
in
this group do not
all confine
their attacks to one sub-group only, but they will be dealt with under the
•heading of that sub-group in which they occasion most damage. A large
number
of these
pests
are cosmopolitan, having been distributed chiefly
by
artificial
agencies
{i.e.,
Corn
Weevils,
Cockroaches,
Rats). Others
have a wide distribution from
natural agencies, such
as ocean currents
{Teredo worms). ANIMALS
DESTRUCTIVE TO
MAN'S BUILDINGS AND LARGER
CONSTRUCTIONS AND
HABITATIONS. ANIMALS
DESTRUCTIVE TO
MAN S BUILDINGS AND LARGER
CONSTRUCTIONS AND
HABITATIONS. Protozoa to Chsefcopoda ... None. Craastacea
Limnoria terebrans and
Zi^/norH?^, and others do damage
to marine works and shipping. Arachnida
None. Hexapoda
Numerous
insects
destroy
the
woodwork
of
bridges,
telegraph
poles,
etc., such as
Termites or white ants;
carpenter
bees
{Xylocoim); death watch
beetles
(.4; io-
hiuvi)
\ ants (ForTnicidae). Chilopoda and Diplopoda
None. MoUusca
Teredo worms damage marine works by boring into the
wood
; Dreisscna by
entering
water
pipes
;
Saxicava
burrow into stone piers. Tunicata
,
None. Eishes
None. Birds
Birds do
damage and cause annoyance by building
in
chimney stacks
(sparrows,
storks)
and
by
destroying
mortar in buildings
(pigeons). Woodpeckers damage
telegraph poles in Germany. Mammals
<
Burrowing animals may undermine man's buildings and
habitations
(rabbits,
rats,
mice),
and dam-forming
animals
(beavers),
by
causing
floods, may
damage
bridges
;
otters, voles, by burrowing, damage canal and
river banks. StTB^GEOTPS C
Alfl) B >0F GrOUP E, It
is
difficult
to enumei-ate
the
members
of
these two sub-groups,
Xumerous parasitic worms attack wild rabbits, big game, and game birds. Mange miles and
ticks
attack
the
fox and other animals. Birds destroy
the
useful
earth-worms. The
schedules
adopted
by the
International
Conference for the Preservation of "Wild Animals in Africa may be referred
to as giving some
indicaitions on the
subject. In Sub-group D we find
numerous insects damaging sucli wild plants as rushes, croci, the cranberry,,
pepper plants, and familiar forest'-trees-already considered. First Report on Economic Zoology. xxxii animals injurious to man's food and other stores. Protozoa to Crustacea
None. Acarina
A few acari attack food—housebold raites (^Olyciphagus)
cheese mites (Tyroglyphus)
; sugar mites (GlycipJiagus). Hexapoda
Numerous insects attack man's food and
other
stores,
both dry goods and fresh—com
weevils
{Calandra)
;
bacon
beetles {Dennestes)
; cheese
fly {Piophila)
; blow
flies
{CallipJuyra)
;
cockroaches
{Blattidx)
;
cigar
beetles (Lasioderma)
; drug beetle {Atiobium paniceum)
;
death-watch (4 fropos divinaUn-ia)
; silver fish (Lepisma). Chilopoda and Diplopoda
None. Mollusca
Slugs attack corks. Tunicata to Birds
None. Mammals
Rats and Mice. Protozoa to Crustacea
None. Acarina
A few acari attack food—housebold raites (^Olyciphagus)
cheese mites (Tyroglyphus)
; sugar mites (GlycipJiagus). Hexapoda
Numerous insects attack man's food and
other
stores,
both dry goods and fresh—com
weevils
{Calandra)
;
bacon
beetles {Dennestes)
; cheese
fly {Piophila)
; blow
flies
{CallipJuyra)
;
cockroaches
{Blattidx)
;
cigar
beetles (Lasioderma)
; drug beetle {Atiobium paniceum)
;
death-watch (4 fropos divinaUn-ia)
; silver fish (Lepisma). Chilopoda and Diplopoda
None. Mollusca
Slugs attack corks. Tunicata to Birds
None. Mammals
Rats and Mice. Introduction. xxxiii xxxiii Hexapoda
continued
books
and
papers damaged
by book-worms
{Atropos
divinatoria) and by
CTielifers
or
tailless
scorpions
in
India,
also
by
Lepisma. Clotbing
also damaged
in
India by Anthrenus vorax. Cbilopoda and Diplopoda
None. Tunicata to Birds
None. Mammalia
Mice and rats (occasionally). Hexapoda
continued
books
and
papers damaged
by book-worms
{Atropos
divinatoria) and by
CTielifers
or
tailless
scorpions
in
India,
also
by
Lepisma. Clotbing
also damaged
in
India by Anthrenus vorax. Cbilopoda and Diplopoda
None. Tunicata to Birds
None. Mammalia
Mice and rats (occasionally). Animals which are known as " Beneficials " on account of their
being
destructive
of
or
checking
the
increase
of
the
injurious Animals classed under Groups D, E, and F. Animals which are known as " Beneficials " on account of their
being
destructive
of
or
checking
the
increase
of
the
injurious Animals classed under Groups D, E, and F. The animals
falling
in
this group, spoken
of
as a rule
as " natural
enemies," are best treated
in connection with the pests enumerated in the
groups D, E and
F. They may be
beneficial,
either by
(i) being
pre-
daceous, or (ii) being parasitic upon the pests of crops, animals, and man. ANIMALS
INJURIOUS
TO
FURNITURE,
BOOKS, DRAPERY AND
CLOTHING. Protozoa to Crustacea
None. Arachnida
Acari (GZJ/c^p7^a<7Ms) spoil furniture and are obnoxious. Hexapoda
Termites or white ants; wood-boring
beetles (Xylobinm
and Anobium)
;
leaf-cutting
bees {Megachile)
; clothes
moths
{Tilled);
Derviestes
beetles attack
soft
goods;
cockroaches
[Blattidan)
attack
boots,
also Anobium; Protozoa to Crustacea
None. Arachnida
Acari (GZJ/c^p7^a<7Ms) spoil furniture and are obnoxious. Hexapoda
Termites or white ants; wood-boring
beetles (Xylobinm
and Anobium)
;
leaf-cutting
bees {Megachile)
; clothes
moths
{Tilled);
Derviestes
beetles attack
soft
goods;
cockroaches
[Blattidan)
attack
boots,
also Anobium; Survey op Group G. Protozoa
None. Porifera to Echinoderma
None. Platyhelmia
Parasitic
cestodes
help
to
keep down
certain
noxious
birds and mammals (rabbits, sparrows, and others). Nemertina
None. Nematoda
Act the same as Platyhelmia. Crustacea
None?. Arachnida
Spiders
by
destroying
noxious
insects;
mites
(Der-
manyssus) by being parasitic on destructive birds. Hexapoda
Many insects prey upon other insect pests. Hover flies
(Syrphidie) and lace-wing flies (Henicrobiidx) feed upon
Aphides
; dragon flies {Odonata) upon mosquito larvae,
butterflies, etc. ; carnivorous ground beetles (Carabidm,
etc.), lady birds (Coccincllidai) upon Aphides and scales. Protozoa
None. Porifera to Echinoderma
None. Platyhelmia
Parasitic
cestodes
help
to
keep down
certain
noxious
birds and mammals (rabbits, sparrows, and others). Nemertina
None. Nematoda
Act the same as Platyhelmia. Crustacea
None?. Arachnida
Spiders
by
destroying
noxious
insects;
mites
(Der-
manyssus) by being parasitic on destructive birds. Hexapoda
Many insects prey upon other insect pests. Hover flies
(Syrphidie) and lace-wing flies (Henicrobiidx) feed upon
Aphides
; dragon flies {Odonata) upon mosquito larvae,
butterflies, etc. ; carnivorous ground beetles (Carabidm,
etc.), lady birds (Coccincllidai) upon Aphides and scales. C First Report on Economic Zoology. First Report on Economic Zoology. Chilopoda
Most centipedes kill noxious ground insects and molluscs. Diplopoda
None. Tunicata
None. Pishes
Many fish prey upon mosquito larvse—carp, etc. Amphibia
Frogs and toads especially, by devouring insects and slugs. Reptilia
Snakes
destroy
insects,
rats,
mice,
and
other
noxious
animals (fer de lance, grass snake, rat or corn snake). Birds
All insectivorous birds and some birds of prey (warblers,
swallows, starlings, rooks, kestrel hawk, owls. Starlings
[Sturnus)
devour
ticks
on
sheep
;
Bwpliaga
or
ox-
peckers the ticks on oxen in S. Africa*). Mammals
Many mammals are beneficial (moles, skunk, hedgehogs,
fox, shrews, various insect-eating bats {yes'pertiliomdai)
and others). * The Oxpeckers do some harm
as
well, for when no ticks are present on the ox
or sheep they will wound the back of the animal, pecking deeply into the flesh. E. EAY LANKESTER. GROUP
E. Animals
which
concern Man
by
causing
bodily
injury
or
disease, both
possibly
of a
deadly
character, to
(A)
his
stock of Domesticated Animals, (B)
his Vegetable Planta-
tions, or (C) to Wild Animals in the preservation of which
he
is interested, or (D) Wild Plants
in the
preservation
of which he
is interested. Eel-worm Disease
in Oats. Some oat
plants, sent by a coiTesponclent
of the Board from South
Tawton, Devon, from
a
field that was seriously damaged,were found
to
be
attacked by
eel-worms
{Tylcnchus
devastatrix, Kuhn). The
popular names for the disease these eel-worms cause are " tulip-root
and
"segging."
This
disease
takes
its
name
from
the
swollen
appearance
of
the
base
of the
stem. This
swollen
basal
part
is
suiTOunded in most cases with contorted shoots
of a pale unhealthy
hue. The minute eel-worms
are found in abundance amongst
the
deformed
shoots
and
in
the
stem
(at
its
base). This
species
of
eel-worm attacks
chiefly
oats,
rye,
clover,
onions,
turnips, but
also
occurs
in wheat,
buckwheat, and
various
wild
grasses,
as
sweet- B
2 First Report on Ecoiioinic Zoology. 4 scented vernal and annual meadow-grass
;
it
is also found
in daisies,
buttercups, and
plantains. Teasels and hyacinths
also
harbour
it,
according to Eitzema Bos. It appears that barley
and
carrots are
free from its attack. Part
of
their
life
is
spent
in
the
soil, and
they can
then be
successfully treated as mentioned below. Peevextion and Treatment. 1. Deep
ploughing
in autumn
;
the
depth
should
be
eighteen
inches. By
this
treatment
the
layer
of
earth
that
contains
the
eel-worms
is buried, and so they are put out of the way of the next
crop. Ploughing
at
a
less depth
does good
if
a skim
coulter
is
attached, but the deeper the land is ploughed the better. 2. On eel-worm land avoid crops in
rotation that are
attacked,
and use those
that
are not
i.e. barley. Clover
after
'•' tulip-root
must carefully be avoided. 2. On eel-worm land avoid crops in
rotation that are
attacked,
and use those
that
are not
i.e. barley. Clover
after
'•' tulip-root
must carefully be avoided. 3. Sulphate of potash on a diseased
field does good at about the
rate of 1 cwt. to the acre. There is not the least doubt that by late autumnal deep ploughing^
by judicious
rotation, and by the use of certain
artificial manures,
the pest
can
be
easily fought with
success. It
is
also
said
that
stable manure should be avoided. There was
also
a single puparium
of the
Frit Fly {Oscinis frit)
found in one plant from this district. (Oscinis frit, L.) (Oscinis frit, L.) Oat plants sent to the Board of Agriculture from the sewage farm
of
the
Croydon
Corporation
were
being
destroyed
l)y
a
small
dipteron, which
has done a
great deal
of damage
to the
oat and
barley crops in the south of England. This small fly is the so-called
Frit Fly, the Oscinis frit of Linnaeus, the Oscinis vastator of
Curtis. The larvse of the
Oscinis feed just inside the crown of the plant,
and in the majority of cases destroy it
; but when they feed between
the outer leaves, as they often seem to do, the crop may survive. Crops
that look irreparably damaged often tiller out and produce
a moderate
yield. This year (1902)
the
pest has
]jeen very harm-
ful. The
flies were nearly all hatched out by June, but some sent
in
the sample
of oat plants from Croydon were only just entering
the pupal
stage. The second
lu'ood
probably lays
its
eggs
on the Reports
to
the Board of Agriculttire. 5 developing grain. In Sweden
tins
second attack
is
often harmful,
producing light shrivelled samples of corn (frits). Little or nothing can be done when
a crop
is badly attacked, as
the one reported from Croydon. Moreover, the damage
is now (June)
done, and the second brood cannot be materially lessened, even by
ploughing up the crop, as they are hatching out rapidly. One
feature
has been
noticed
in
districts where
Oscinis frit
is
abundant,
namely,
that
early
sown
crops
suffer
the
least. Oats
sown 29th of March were not attacked
; those sown on 29th of April
had over 70 per cent, of the stems attacked. Where
a
field
is ureparably damaged
it
is
as
well
to
deeply
plough
it up
at
once, so as to bury the puparia
of the
fly and thus
prevent
the second and summer
brood, that
cannot
be
so
easily
destroyed, from appearing. Wlien early signs of the crop being attacked are
noticed, the loss
may
be
materially
lessened
by
the
timely
application
of
some
stimulating dressing such as nitrate of soda. Sil'pha
riigosa,
L., on Turnips, and other Silphidw. Sil'pha
riigosa,
L., on Turnips, and other Silphidw. One
of the
Carrion
Beetles, Silpha rugosa, L., closely related to
the Beet
Carrion
Beetles (Silpha opaca and
>S'. atrata), sent
to the
Board, was appearing in great nninbers on a turnip
crop. Both of
the
latter species
are injurious
to mangolds,
the
larv?e devour
the
leaves and stem and often destroy
a crop
entirely,
Silplia rugosa
occurs with
them
nearly always
and
is
probably the commonest
member of the genus, being found almost everywhere, but there have
been no observations made on this
species showing that it does any
harm
to
crops, such as
occurs with
the two closely related Carrion
Beetles. The
larviie
of the
Silphidiie
and
also
the
adults
are
normally
carrion
feeders. Numbers
of
these
three
species may usually be
found in June
in and under any dead carcase laying about in the
fields, but as previously stated the two species S. opaca and
>S'. atrata
frequently give up theii' carnivorous habits and attack mangolds and
beetroots. Canon Fowler also records finding the larv?e of a member
of
this genus on the
roots
of plants
in
the
Isle
of Wight. It
is,
therefore, possible that
S. rugosa may sometimes become
vegetarian
in habit, but no mention has been made
of
this, nor does the Board
correspondent make any such statement. The larvre of S. rugosa are moderately broad with the edges of the
thoracic
(i.e. first
three) segments rounded, those of
the remaining
segments
projecting. The
l)ody ends
in two processes
called
cerci,
which in
this
species
are
long, at
least
tlu'ee times
as long as the
anal process between them
;
the head is large and projecting. When full grown, which is usually by the middle, but sometimes
not
until the end, of July, they bury themselves in
the
soil
to a
depth of from three to four inches and turn to pup?B, After from three
to four weeks beetles appear from these pupse,
and these beetles apparently live through the winter. Miss
Ormerod
records
the
Beet Carrion Beetle
{S. opaca)
on
potato and
also
as devouring the
Spurrey
(Sjyergula
arvensis), and
it
is thus possible
that we may get
S. rugosa working in a similar
manner on various plants and not on any one in particular. Smut in Barley and Insects. The barley sent by a correspondent
of the Board from Brackley,
Northamptonshire, was found to be suffering from the fungoid disease
called Loose Smut
( Ustilago nuclei, Jensenii). Two methods
of treatment are known for Smuts
:
(1) the " blue-
stone " treatment and
(2) the " hot water "
treatment. The former
does well
for covered smut
( Ustilago jensenii, Eostr.), but has
little
effect on Loose Smut. The " hot water" treatment consists of soaking the
grain for five
minutes in hot water heated to 126° F. The
grain should be warmed just before by putting it in a sack
and steeping
it
for a few minutes in water of 120°
F. After the
erain has been
five minutes
in the hot
water, 126° F., it should be
taken out and plunged
right away into cold water and then spread
out to dry. Some Clavicorn Beetles
of the genus Phalacrus of PaykuU were
found in the diseased ears. They apparently feed upon the spores of
the Smut fungus, but
at the same time they no doubt help to dis-
tribute this fungoid disease, for they are often seen covered with the
spores, amongst which they crawl during feeding time. The remedy
of this combined attack lies in treatment of the seed
; no steps need be
taken in regard to the insects. First Report on Economic Zoology. First Report on Economic Zoology. 6 Sil'pha
riigosa,
L., on Turnips, and other Silphidw. As
there
is
a
possibility
of
these
carrion
beetles
becoming
destructive, steps should l)e taken to have the turnip and other fields
cleared of them and any of their
larvse. To do
this
is a compara-
tively easy matter
if we employ natural traps
in the form
of dead Rep07'ts
to the Board of AgiHctdture. 7 birds
or small mammals, scattered over the ground every here and
there. The dead bodies attract the beetles and the larvte, which can
be shaken out into pails of hot lime and so destroyed. Caterpillars
(Surface
larvce) on Turnips,
etc. The Turnip
or Dart Moth {Agrotis
segetis) and the
Heart and Dart Moth
{A. exdamationis). Caterpillars
(Surface
larvce) on Turnips,
etc.
The Turnip
or Dart Moth {Agrotis
segetis) and the
Heart and Dart Moth
{A.
exdamationis). Some
larvai attacking turnips and
potatoes, sent by a correspon-
dent of the Board of
Agriculture, were the caterpillars of the Heart
and
Dart
Moth
{Agrotis
exdamationis),
popularly
called
Surface
Larvffi—Cutworms
in Canada and the United States. Two
species
occur in abundance, viz., A. exdamationis and A. segetis ; the latter
is called the Turnip Moth. Another correspondent, writing from Dadlington, Nuneaton, states
"that
turnips
are eaten
off and
potatoes burrowed
into. In
this
district acres are spoilt by eating the turnips under the ground." These
caterpillars
were
also
sent
by
a
correspondent
from
Loughborough, where they were attacking mangolds. There
has been
a
serious outbreak
of
these
pests
during
the
present year in all parts of England and Wales. A leaflet (No. 33) has been issued by the Board on
these pests
under the
title of " Surface
Caterpillars."
To this leaflet the follow-
ing information should be added
: A leaflet (No. 33) has been issued by the Board on
these pests
under the
title of " Surface
Caterpillars."
To this leaflet the follow-
ing information should be added
: («) No
dressings can be applied in
sufficient
strength
to
kill
these
caterpillars
as
recommended, but
the
dressings
do
good
in
helping on growth of the plant. («) No
dressings can be applied in
sufficient
strength
to
kill
these
caterpillars
as
recommended, but
the
dressings
do
good
in
helping on growth of the plant. (h) Land
after
mangolds
had
best be
lightly
broken
up, not
deeply ploughed
;
this would bury, not expose,
the
larvaj, and
bii'ds
are the greatest help in keeping them in check. (c) In land invaded by Cutworms, a crop of must'ard ploughed in
has done good. {d) The long and exhaustive series
of experiments, conducted at
the Agricultural Experimental
Station of Cornell
University, have
shown that by far the most
successful way of combating these pests
is the employment
of
" poison -baits."
These "poison-baits"
have
been
employed
in
the
field
by
soaking
clover,
lucern,
etc., in
a
solution
of Paris green and throwing
little heaps of
it about in the
field amongst the roots,
etc., or, in the case of garden cultivation, bran
soaked in Paris green may be used. When
clover,
etc., is employed the
solution
of
Paris
green
in First Report on Economic Zoology. Caterpillars
(Surface
larvce) on Turnips,
etc.
The Turnip
or Dart Moth {Agrotis
segetis) and the
Heart and Dart Moth
{A.
exdamationis). 8 wliicli
it
is
dipped
should be one pound
of Paris
green
to
tifty
gallons of water. For bran
bait use 1 lb. of Paris green
to 25
lbs. of wheat
bran,
and mix with just enough water to make a mash. Of course it should not be placed where poultry go
or sheep or
other stock feed. In
mangold
lields
the
clover
bait
might
be
tried
where
the
attack is very bad. The fresher the clover the better the results would be. The baits
should be placed on the ground late in the afternoon. The
green-stuff might
be
sprayed
with
the Paris green
l^efore
being cut—so as to save the trouble of dipping it. The Pigmy Mangold Beetle.
(Atomaria
linearis,
Stephens.) Some
beetles
sent by a correspondent of the Board from Barn-
staple
that were destroying his mangolds proved
to be the Pigmy
Mangold Beetle {Atomaria linearis). A
similar attack was reported
to Wye College by Mr. Thos. Powell. In
this
case
the
beetles had
destroyed
a
field
of
mangolds
on
the Waldershare
estate. Miss
Ormerod has recorded damage
to mangolds by an Atomaria which
she identified as liiicaris, Stephens. As far as
is known, these
little
beetles
(Fig. 1,
d)
destroy
the
sprout
of
the mangold
seed
just
as
it germinates, and
later they
attack both root and the leaves. The leaves
are gnawed away and gradually die (Fig. 1, c)
; they
also gnaw away
the lower parts of the leaf stalks below the ground
level and
so
kill the plants. The tap root is attacked, the damaged
part turning black (Fig. 1,
b). It thus
seems that all parts
of the
plants
suffer
in
the young
stages
and
during
germination. The
beetles may be found in great numbers on the ground under clods
of
earth, on
the
leaves and
in
the earth around the
roots. They
appear in May and June and seem to decrease in July and August,
those
occurring
in
the
last two months apparently being a second
brood. Nothing
is known of
its
life-history,
but
apparently
the
larvae feed below ground, probably on the roots of the plants. This
attack was
first observed
in 1839 by M. Bazin
at Mesnil
St. Firmin, and later Macquart noticed
this pest devouring the fields
of red beet in the environs
of Lille
to such an
extent
that whole
crops were ploughed up. It occurs in Great Britain in many districts
in great numbers and does much harm
;
it is not noticed on accoun Reports
to the Board of Agriculture. 9 of
its
small
size. It
seriously injured the mangold crop at Ciren-
cester in 1891
;
it is also recorded as damaging mangolds at Lyming-
ton, Ashburton (Devon), Weston-super-Mare,
Sliifnal
(Shropshire),
Denham (Bucks)
; and
it has been abundant
this year in the neigh-
bourhood of Wye, Kent. The
beetles may be noticed on the wing,
pairing on warm
evenings. They probably hibernate
in the adult
stage. Fig. 1.—THE PIGMY BEETLE {Atomaria
linearis). A, damaged plant
;
h, damaged tap root
;
C, holes eaten in leaves
;
D, adult. Fig. 1.—THE PIGMY BEETLE {Atomaria
linearis). The Pigmy Mangold Beetle.
(Atomaria
linearis,
Stephens.) A, damaged plant
;
h, damaged tap root
;
C, holes eaten in leaves
;
D, adult. The
best way
of
destroying
this
pest
would
be
to
run
a
Strawsoniser over the lield with ordinary Paris green wash in July,
when the beetles seem to feed mainly on the young seed leaves. The
best way
of
destroying
this
pest
would
be
to
run
a
Strawsoniser over the lield with ordinary Paris green wash in July,
when the beetles seem to feed mainly on the young seed leaves. If the crop
is
destroyed
as
is
the
case
with
this outbreak
at
Barnstaple, deep ploughing would be advisable. In
districts on the Continent where
this
beetle is a serious pest
to
sugar-beet, thick sowing
of seed is practised and would be worth First Report on Economic Zoology. lo (loiiiiT;
in
the case of mangolds in
this country in districts where the
beetle occurs in large numbers. If
the land
is
in good heart maize may be put in
in
the
place
of the
mangolds
that
have
been
destroyed. The
beetles would
probably leave this plant alone, if they are not all destroyed by deep
ploughing. This
is not an uncommon
pest,
liut
is not reported very often
on accoimt of the damage
being
attributed
to
other
causes, such as
ants,
etc.,
the
beetles, owing
to
their small
size,
being
either not
observed or if observed mistaken for ants. {Aj)liis atriplicis, Linn.) Some
insects
attacking the seed heads of the mangold were sent
to the Board
of Agriculture from Childerley Hall, Cambridge
; they
were one
of
the Aphides known
as
Ajjhis
atriplicis,
Linn. This
species
feeds normally on the Chenopodiums in
the summer and on
the common Orache (Atriplex 2)cdula) in the autumn. The apterous females
are
of various
colours, green, olive
green,
black. Buckton
descril^es
four distinct
varieties
:
(1) wholly black,
(2) black with orange
tibite and white patches on the body, (3) body
green with white
bands,
legs
ochreous
or
whitish,
(4) head and
thorax
black, abdomen green with white
spots. The pupal
stage
is
black with white patches, thorax and wing cases olive. The wdnged
female that produces living yonng is dark olive, the abdomen
barred
with black and with
lateral
spots
;
the honey tubes are green
at the
base and
black at the apex
;
the
legs
are yellowish except
the hind
femora and
tips of the other femora. The male is wingless and of a
greenish-yellow
colour,
head
black,
thorax
with
black
markings
the abdomen
has
three
longitudinal rows
of
black
spots
forming
almost bands on the apical part. Legs, and cornicles dull grey. The oviparous female is also apterous and
green, tlie head having
two dark spots. Flies (Bibiomdce) on Mangolds. Some
flies were sent by a correspondent from
Billericay of the
dipterous family— Bibionida?, species
Bibio Jioriulanus—from a
field
of mangolds cleared
oft' in one night. These
flies can have had nothing
to do with
the
two
acres
of
mangold reported to have been destroyed. Although
their
larvte
are
more
or
less
injurious
to
roots, the
adult flies do no harm, not having a biting or piercing mouth. The damage reported seems
to point
to the small
beetle recently
sent
to the Board from North Devon—the Pigmy Mangold Beetle
{Atomaria linearis), which is evidently abundant in some parts. The correspondent was advised to look
for
these small
beetles,
which may be most easily caught by pulling up the young mangolds
and the earth round them, when the beetles fall out of the soil. On
fine days they occur above ground as well {vide page 8). In any case the flies sent cannot have damaged the mangolds in
the way reported. The Life-histoky. Little
is known concerning
its
life-history. The females of the
last
generation
lay
their eggs on
the dead
rolled up leaves of the
plants upon which
they have
been
feeding, amongst
their
debris
formed
of cast
skins, frass, etc. The ova are elongated
oval, yellow
at
first, and
gradually become
black. These eggs
are
laid
in
the
autumn
after
the apterous males have appeared and
fertilised
the Reports
to the Boai'd of AgricultMre, 1 females. How the wiuter
is passed
is
not
known. In the
early
and
late summer they
feed upon wild Chenopodiums and mangolds
and in the autumn on Afrijdcx latifolia. Not only does this aphis cause the leaves to roll up longitudinally,
but they also feed upon the seed heads of the mangold. Spraying
M'ith
paraffin
emulsion
or
quassia wash would check
their increase and clear most of them
oft'. This, of course, should not
be done when the sun is out. First Report on Ecoitomic Zoology. First Report on Ecoitomic Zoology. 12 Tkeatment. Soot and lime have been found of some benefit. The chief thinff
to do, however, is to treat the land with gas-lime during the autumn
and winter
to
kill the hibernating
insects, then
in
the
puparium
stage in the soil, and so prevent their doing damage another year. On
the
Continent
some
good
is
said
to
have
been done by
applying superphosphate of lime as a preventative. Treatment
with gas-lime
is
the only certain way
of
lessening
these pests on a large scale. Tlie three small caterpillars were probably non-injurious. Tlie three small caterpillars were probably non-injurious. Muscid Larvae attacking Roots. A correspondent
of the Board of Agriculture
sent from
Burley
Beacon, lUngwood, Hants, two
different
kinds
of
larvae
attacking
roots of plants. (1) A dipterous maggot changed during
transit
into the so-called
chrysalis
or puparium
stage
of
one
of
the
flies
belonging
to
the
group Antlwinyida:. There are
several of
these
diptera which are
root-feeders
in
their maggot
stage. Probably
the
one
sent
was
Antlwmyia radicum, L. (1) A dipterous maggot changed during
transit
into the so-called
chrysalis
or puparium
stage
of
one
of
the
flies
belonging
to
the
group Antlwinyida:. There are
several of
these
diptera which are
root-feeders
in
their maggot
stage. Probably
the
one
sent
was
Antlwmyia radicum, L. (2) Three small larvaj which were the
caterpillars of one of the
Tinese. (2) Three small larvaj which were the
caterpillars of one of the
Tinese. The damage was probably
all caused by the Anthomyia
larvse,
which are often serious root pests. {Mdolonthidcc. ) Several enquiries have been made
at
the
Board
of Agriculture
during
the
past
summer
concerning
Chafer
larvte,
the
so-called
White
Grubs. From
Ferryside,
South
Wales,
the
grubs
of
the
Garden Chafer or Cock-y-bonddu (PA?///ojjer^7i« horticola, L.) were sent
on
June
20th. They
were
leported
as Antler Moth
caterpillars
(Charwas graminis, L.). The Cock-y-bonddu is apparently the common
Welsh
chafer, for nearly all the
larva3 examined have proved to be
this species
; the attack seems
to have been fairly general in Wales
this year. The Summer Chafer {Rhizotrogus solsiitialis, L.) has
also
done much damage in many parts of Great Britain. From Launceston
they were
especially reported
as damaging
pasture
land,
also from
Wye. By far the most abundant
species, however, has been the Cock-
chafer {Mdolontha vulgaris, Fabr.), which has occurred this summer
in
large
numbers
in
the
following
localities
:
Wetherby,
York,
Chester, Lytham,
Southwell, Pewsey, Eoydon, Limpsfield, Croydon,
Tooting, Highgate, St, Leonards, Catford and AVye. The brood appeared from the first to the thiixl week in June. The
Summer Chafer {R. solsiitialis) has occurred as adults
at Wye,
the
brood occurring from the
third week
in July to the end of August. In
districts
whei'e these broods
are
recorded we
shall now know
when to expect the next brood of beetles and so be prepared to take
steps to collect them wholesale as soon as they appear. In this way
only can any real good be done in districts where these Chafer larvae
are
harmful. Full information
regarding
these
pests
is given
in
the revised leaflet No. 25 of the Board. Reports
to the Board of Agricidtttre. 13 Leather-Jackets or Larvae of
Tipulidce. The
insects sent
to the Board of Agriculture by a coiTespondent
from Eaton, Norwich,
that had been damaging the
roots
of
grass
proved
to be the pupre
of one
of the Daddy Long Legs {Tipulidse). The larvie of these Tipulidse are known as " leather-jackets," and are
very destructive to all kinds
of roots, especially grass. Pasture land
is often ruined by them. They were those
of the Yellow Spotted Crane Fly (PachyrJiina,
maculosa), whose
larvoj work
in a very similar way to those of the
Common Crane Fly (Tipula
oleracea). The
pupa3' of the latter are
larger than those of the former. (A full report on these pests is given on pages 94 to 104). (A full report on these pests is given on pages 94 to 104). The Green Rose Chafer {Cetonia
aurata, Lmn.) on
Beans and Currant Bushes. The Green Rose
Chafer (Cetonia aurata) was
sent to the Board
from
Gloucester,
with
a
note
that
they were appearing in
great
quantities and were stripping the beans and currant bushes of
tlieii'
leaves. This
beetle
is generally distriljuted
in the South of England and
occurs
in plenty in the Midlands, but becomes
rarer in the North. It is usually very common at Gloucester, so that
it
is not surprising
that it now and then occurs in such numliers as to become a
serious
pest. Tlie beetle attacks
all kinds of flowers and also the leaves
;
it
is
especially injurious to
tlie
rose, apple and strawberry. It is also
recorded
as damaging turnips
for
seed. When
attacking
blossoms
the beetles seem to mainly devour the anthers and thus destroy the
crop. They are very frequently found in Peonies and on the Elder
they also destroy Iris blossom
at
times. These
l3rillant
Ijeetles
fly
readily
in
bright sunshine, but become very
.sluggish
during
dull,
damp, and cold weather. LiFE-HISTOEY. The
beetles appear from
the middle
of May on through
June. They lay their eggs in the ground, seeking out some crack or crevice
into which they crawl. Heaps of rich earth such as cucumber beds
and vine
borders
are
favourite places
for them
to
lay
their
eggs. These soon give rise to white grubs very like those of the Cockchafer, First Report on Economic Zoology. 14 but which can easily be tokl by having a deep reddish-brown spot on
each side
of the first thoracic segment
;
the legs are also longer than
in
the
Cockchafer
grub,
and
the
whole
surface
is
clothed
with
transverse rows
of
reddish-brown
hairs. The
larvaj may
also
be
found amongst rotten and rotting wood, but mainly in rich soil
; their
food consists
chiefly of the
roots
of various
plants and probably of
decaying vegetable matter as well. When full grown they attain the
length of an inch and a
half, taking from two to three years to reach
maturity. The pupal stage takes place in an eaithen
cell over an
inch in length formed deep in the ground
; the outer part of the cell is
rough, the inner surface smooth. The pupa
is
of an ochre
colour. A
B
Fig. 2. THE GREEN BOSE CHAFER
(Cctoilia aurato). A, Imago
;
B, Larva, B Fig. 2. THE GREEN BOSE CHAFER
(Cctoilia aurato). A, Imago
;
B, Larva, The grubs appaiently pupate in the summer, and the beetles appear
from these in the following May and June. Canon Fowler notes that
the little larv<Te and perfect insects are often found in ants' nests. Myriapoda in Potatoes. Numbers
of centipedes and some millepedes
were
sent
to
the
Board from Honiton with a note
to the
efiect that they (the centi-
pedes) were destroying the potato crop in that neighbourhood. The
Scolopcndridse were mostly
alive
in
a
small
tin
box, but
several had been killed by the stronger ones in the box. These Scolopendiidte are certainly carnivorous and do not seem
to be destructive
to
roots, although
Curtis mentions
such
a
habit,
quoting the following from a
correspondent
:
" Mr. Hope attributed
the potato disease
to the
attacks
of wire-worms, and also to a small
Scolopendra which was found in myriads
infesting
diseased potatoes
at Southend." In
all
cases
where
these
myriapods
are
sent
as
the
culprits
other pests will be found on
careful examination. In the box sent
from Honiton were
also the remains
of some small
Julidce
which
have undoubtedly been the cause
of the
trouble. The large centi-
pedes sent had probably been feeding
off
these
Julidoe and
other
animals in the soil. With
regard
to destroying the
Julidte, nothing further can
be
added to the information given on pages 86 and 105. Remedies. By far the most successful way to cope with these large sluggish
beetles
is by
" hand-picking."
This
should
be
done
during
dull
weather when they are very quiet,
as on warm
days
they become
more active and fly about. Heaps of leaf mould, cucumber beds, and heaps of decaying wood
should be examined when turned over or moved and the grubs hand-
picked. Old
tree stumps frequently harbour them and should thus
be
grubbed
up
in
the
winter
and
burnt. In
garden
and
field
•cultivation poultry do much good if turned on
to the land when it is
being broken up, for they greedily devour these larvse as well as those
of the Cockchafer. " Tui'f-traps,''
i.e. heaps
of
rotting
turf, may be
left
here
and Reports
to the Board of ^Agriculture. 15 there about
in the garden
to
attract
any
stray
beetles
to
deposit
their eggs
; these heaps can be examined in the winter and all the
grubs burnt. Wire-worm (Lacon murinus,
L.) in Potatoes. An
insect
sent
to
the
Board
in
a
potato
from
Barley,
near
Burnley, proved
to be the larva
of one
of
the
Elateridpe
or Click
Beetles,
i.e., a wire-worm
Lacon muriiius. There
is unfortunately no remedy when wire-worm get
into the
potato crop. The
field should be deeply trenched
later on and
a
crop of mustard grown afterwards. Sometimes wire-worm will leave
potatoes for wurzel and
caiTot, so that slices of either, if procurable,
might be put here and
there along the rows just under the ground
and examined every few days, or rape cake may be spread between
the rows, as
this
class
of larvte
are very
fond
of this
as
food, and
would probably be drawn away from the plants. (Mr. Deadman, of Wye, finds that beet-root forms a much more
attractive bait than any other root for catching these pests.) (Mr. Deadman, of Wye, finds that beet-root forms a much more
attractive bait than any other root for catching these pests.) First Report on Economic Zoology. 1 6 A New Potato Feeder
The Cinnabar Moth {Euchelia jacohecs). During the past year quite a number of new potato pests have
appeared. Amongst them may be mentioned the
caterpillars
of the
Cinnabar Moth
(Uuchelia jacohecv),
sent hj a correspondent
of the
Board from Alton, Hampshire. This moth
is
fairly common. The
front wings when expanded measure from an inch and a half to an
inch and three-quarters across
; they are dull black with a narrow red
stripe near the upper margin and two
spots on the outer margin of
the same colour
; the hind wings are
scarlet red with a nan^ow dull
black margin. The moth appears
in May and June and flies slowly
during the day-time. The caterpillars
feed usually on the Eagwort
(Senecio jacohcva), and sometimes, as stated by the Board's correspon-
dent, on
the
Groundsel. As
far as can be found out the potato
is
quite a new food-plant. As the larva? are found
in companies, they
could easily he cleared out of the potato crop, which should certainly
be done, as they are very ravenous feeders. Eagwort may frequently
be seen quite stripped of its leaves by these black and orange ringed
larvie. The Mustard Beetle (Phcedon
hetulce,
Linn.), The Mustard Beetle (Phcedon
hetulce,
Linn.), The Mustard
Beetle
{Phcedon
hetulce,
Linn.) was
reported
this
year (1902) to the Board as very destructive at Holbeach, Lincolnshire,
and information was asked for as to the best plans of coping with the
attack. It can be materially lessened by various methods. The beetles pass the winter in a torpid condition
in any shelter
where they have been working in the summer and autumn. The larvic hatch from eggs laid in the spring upon various plants. The
beetles which deposit
tliese spring eggs have previously passed
the winter in hollow stems
of reeds along the dykes and ditches of
the
district and
also commonly in the hollow mustard
stocks
left
about in and around the fields and also in the mustard stubble. They
also winter in mustard
stacks,
cracks and
crevices
of gates, posts,
fences, rough
grass
and
all
manner
of
places. The
larva? when
mature pass into the ground to pupate, in which stage they remain
from two
to
three weeks
;
the
beetles coming
from
these pupa? at
once attack the mustard crop. We find the beetle practically all the
summer
:
it
is therefore probable that there
is more than one brood Reports
to the Board of Agriculture. 17 every year. The eggs
laid iu the spring are placed on
all kinds of
Cruciferaj
;
the
larvfe feed upon
the
leaves
;
they
are
dull, smoky
yellow creatures, slightly hairy and spotted with
l)lack, the liead and
the
six
legs are also black
;
there
is
also a distinct caudal foot and a
row of tubercles along each side from which can be protruded curiou*
yellow glands
; when mature they reach
al)out
three-fourths
of an
inch in length. These
larvai can be
easily seen on
the
leaves, and
are vulnerable at this stage. PrEYENTIYE and EEMEDIAL MeA8U1!ES. All precautions should be taken to destroy as much winter shelter
as possible. After a bad
attack
it would be advisable
to burn the
mustard straw, not
at
once, but
after
it has been allowed
to stand
some time in heaps in the fields
; the beetles would seek winter shelter
there, and on firing the heaps they would be destroyed. All hedge trimmings and reedy growths along dykes should be cut
and burnt during the winter. No experiments on
a
large
scale
seem
to have
been made in
destroying the larvaj upon the young plants when it
is posdble to get
on the land. There is no doubt that the proper time
to attack
this
pest is in its larval stage when feeding upon the y<ning leaves. The fields should then be sprayed by means of a horse Strawsonizer
with
Paris-green wash
; the time
to
carry out
this
operation would
depend upon the time the grubs are noticed on the leaves. The
beetles
also
attack
the
young
leaves, and would
also
be
destroyed by the same wash. The beetles may also be collected, when
present in numbers on the young plant, by dragging a long
strip
of
tarred sacking attached
to
a
light rod over
the
fields, and
also by
special machines. The beetles which attack the crop later on
in the
year may be kept in hand by preventing their movements from place
to place. Towards the latter part of the year when
so much damage
is reported, the l)eetles do not seem inclined to use
their wings, but
migrate
in a
l)ody along the ground from
field
to
field. Tliey can
thus be " held up " like locusts by cutting a trench across their line of
march, or by burning damp straw so that the smoke blows on to them. The employment
of a shallow trench about
a
foot deep
is the best
plan to check them, especially if it can be filled or smeared repeatedly
with tar. It
is
also important
to
keep
horse-hoeing
as
long
as
possible
between the rows
; by
this means the pupa? are turned out
of
tlie
earth and are exposed to the attack of various birds. c 1 8 First Report on Economic Zoology. Mustard should always therefore be drilled far apart when grown
for seed
; more than a foot should be allowed between each row. An Enquiry re Bud Mites {Eriopliyes
ribis^ Nalepa) in
Black Currant Bushes. A correspondent
of the Boai'd
of Agriculture living
at Suckley
•sent an enquiry regarding
the
stacking
of
diseased black currant
bushes and the subsequent escape of the Bud Mites {Eriophycs rihis). Information was
sent that
it
is
best
to burn
the
cuttings from the
Wack cun-ant bushes
infested with the " Big Bud " mite. There are
several points not yet
settled in the life-history of this
pest, notably
how long the egg stage
lasts
; under the circumstances,
it
is
best to
destroy all infested parts, as ova will be found
at most times of the
year in the buds. The probability is, however, that if the black currant faggots were
stacked in the centre of the rick
of other wood, that the acari w^ould
die
out and
the
eggs become
destroyed
;
but, owing
to
the
great
increase and ravages
of this pest, it is best to be on the safe side and
to recommend the burning
of the
infested cuttings, which cannot be
of much value as wood. The most complete
life-history of
this
serious
pest has
recently
appeared in the Journal of the S. E. Agricultural College, by Mr. E. J. Lewis (No. 11, pp. 55 to 80 (1902)). PrEYENTIYE and EEMEDIAL MeA8U1!ES. Not
only can the crop then be easily horse-hoed, but special machines can
be taken
across
the
fields
between
the rows
to
catch
the
beetles. Wooden scoops, with
tar or
soft soap smeared over the insides, may
be arranged
so
as
to be pulled through
the
field, either by hand or
horse-power, and so collect the beetles. The
early
spraying with some
arsenical wash
so
as
to
kill the
larvfE and beetles
is, however, most to be recommended. The Apple Blossom Weevil. (Antho7iomits ^^omoram, Linn.) (Antho7iomits ^^omoram, Linn.) Some apple blossoms sent by a correspondent
of the Board from
Cottenham
on
July
20th
were
attacked
by
the
Apple
Blossom
Weevil (An/honoimcs poonorum). The
blossoms
all
contained
the
mature
beetles, ready to emerge. These beetles feed upon the leaves Reports
to the Board of Agricultitre. 19 to some extent
for the
rest of the
year, and
hibernate
during
the
winter under the bark of
trees, and amongst rubbish, etc., at the foot
of hedgerows. It appears from observations made on the specimens
sent from
Cottenham
that
the
beetles
do
not
leave
the
dead blossoms
for
some days. Much good may be done l)y collecting, as
far
as
possible, all the
dead blossoms beneath the trees (first seeing that all " capped" blossoms
have fallen
;
if not, shake the remainder
off the
trees) and burning
them. At
the same time spray the ground beneath the
trees with
.strong soft soap and paraffin wash, adding double the amount of paraffin
usually employed. At present
all we can do in this
attack
is
to destroy the beetles
and so prevent theu- increase. This can be done in three ways
: (i)
by
that mentioned above
;
(ii) by destruction
of winter
shelter by
use
of caustic
alkali wash
; and
(iii) by jarring
tlie
tiees when the
l)lossom appears in spring so as to shake
off the weevils on to sheets
spread beneath the
trees, when they can be swept up and destroyed. Warm days should be chosen
for this, preferaldy with a S.W. wind. This
has
been
found
to
do
considerable
good
where
properly
-carried out. Stress should be laid on the destruction of fallen diseased blossom. A few days, or even hours, may be sufficient for the beetles to escape,
.and so give them every chance to continue their work next season. Strawberry Beetles. A
correspondent
of
the
Board
sent an enquiry in September
asking
for
information
concerning
beetles
that
had
been
very
harmful amongst the strawberries in parts of Norfolk. The following
report was sent in retuin
: Several species
of ground beetles
attack
strawberries, including
the
following
: Harpahis
ruficornis, Fabr. ; Omascus vulgaris, Linn. Sterojms mandidus, Fabr. ; and Calathus cisteloides, Panzer. These
beetles attack the benies
at
night, usually just when the
fruit
is
ripening. The
insects
remain under
the
earth,
straw,
or
grass between the rows during the day, making holes in the
soil and
having regular runs opening through the
litter. Green fruit
is also
attacked,
the
skin being eaten
away,
the
seeds
usually being
left
intact. There are nevertheless records of the seeds also being eaten,
the ground being described
as "covered
with
a jwwdery
deposit,"
^caused by the seeds eaten off the berries. c
2 First Report on Economic Zoology. 20 The most destructive species appears
to be H. ntficornis, which
is winged, and which evidently migrates in large numbers. These
beetles will feed on other substances, such as live worms,
cooked and uncooked meat,
etc. Harpalus
ruficornis
and
others,
have been recorded before in Norfolk, namely at Walsingham. (Eriocamjm- limacina, Cameron.) The
larvre
of the Tear and Cherry Sawfly {Eriocampa limacina)
have been received by the Board of Agriculture from Enfield. They
were reported by the correspondent as doing damage to fruit-trees and
to
the
hedgerows. They
are frequently very
destructive
to
pear,,
cherry, and other leaves {vide p. 72). Another correspondent from Willingham reports them in September
as damaging the leaves of cherry trees theie. The
life-history,
etc.,
of
this
pest
is
dealt
with
in
the
revised
leaflet No. G2. It has
not
been
nearly
so
abundant
as
in
past
seasons. TREATMENT. The
only successful
plan
is
that
adopted
by
^Messrs. Laxton
Bros., namely, to sink small pudding basins in the
soil between the
plants every few yards and baiting them with
" lights " and sugar-
water
;
the beetles swarm
to
this and are unable
to crawl back up
the
sides
of the
basins
;
similar good
results have been gained by
using
ordinary jam
pots
or
glass
jars. l*robably
poisoned
baits. would act well, Ijut trapping as given abo^'e is a well
tried and most,
successful plan. Maggots in Imported Apples. Some larvie sent by a correspondent
to the Board of Agricultm-e
from Smithfield Market, Manchester, proved to be those of the Codling
Moth {Carpocapsa 'pomonclla). The
correspondent
pointed
out
the
danger
of
constant
fresh
importations of this pest from aljroad. Some observations have been
privately made which couflrm this report. The Codling Moth has been distributed to
countries where it was
formerly unknown in the way described by the correspondent
of the
Board. If large numbers occur alive, as recorded, there is undoubtedly
much harm being done, and this may account for tlie great increase of
this apple pest during recent years, which
in some
cases during the
past season has completely ruined tlie apple crop. As
the
pest
is
very
abundant
in America, and probably
the
unsound apples are shipped to the English market, some steps should
be
taken, if
it proves
to be
a
general
rule, to
safeguard the apple
growers
of
this
country
by
similar
means
to those
employed in
Tasmania. Apple
barrels
have
been
examined
and
numbers
of
Codling
Maggots \\QXQ been found. These get distributed over the country,
and cannot but help increase this pest in our orchards. The maggot in Lisbon apples may prove to be one of the fruit flies
iTrijpda or Ccratitis). Reports
to the Board of Agricttltttre. 21 Small dipterous maggots have
also been recorded—possibly
the
larvffi of the Apple Fruit Fly {Trypeta pomondla), an introduced pest. Small dipterous maggots have
also been recorded—possibly
the
larvffi of the Apple Fruit Fly {Trypeta pomondla), an introduced pest. The larvie of the above can be told as follows
: (a) Codling
^laggot,
pink, with
six
jointed
legs
in
front,
four
pairs of prolegs in the middle of the body, and an anal pah*. (a) Codling
^laggot,
pink, with
six
jointed
legs
in
front,
four
pairs of prolegs in the middle of the body, and an anal pah*. (h) Weevil ^Maggots, white, footless, more or less curved, and with
a wrinkled skin. (h) Weevil ^Maggots, white, footless, more or less curved, and with
a wrinkled skin. (h) Weevil ^Maggots, white, footless, more or less curved, and with
a wrinkled skin. (c) Sawfly Maggot, white, six jointed legs in front, more than four
pairs of fleshy prolegs. (c) Sawfly Maggot, white, six jointed legs in front, more than four
pairs of fleshy prolegs. {d) Fruit Fly Maggot, white, footless, not curved, small. Fruit Fly Maggot, white, footless, not curved, small. {d) Fruit Fly Maggot, white, footless, not curved, small. Maggots in Apples. A con-espondent from Uttoxeter ft)rwarded to the B(jard an apple-
that had been damaged by the Codling Maggot {Carpocapsa pomoneUa^
Linn.). This pest is dealt with in Leaflet 30. The correspondent refers to the damage to the apples being due to
weevils
:
" We are suffering in our trees from the
effects
(jf a weevil
that has spoiled and lost us most of the fruit." Two
weevils have been
recorded attacking
apples, namely, the
Purple Apple Weevil
(Ehijnchitcs hacckus,
Linn.) and
the Copper
Coloured Weevil (B. cuprcus, Linn.). Apples also suffer from another grub which has been very preva-
lent this year, the Apple Sawfly {Hoplocampa festudiuca, Klug.). Reports
to the Board of Agricttltttre. Infestation of Fruit Trees by Winter Moth
Caterpillars,
etc. Winter
]\Ioth
{Chcimatohia
hrumata,
Linn.)
caterpillars
were
reported
destroying the
foliage
<jf fruit trees by a
correspondent
of
the
Board
of
Agriculture
at
Laceby,
near
Grimsby. Advice
as First Report on Economic Zoology. 22 regards spraying with
Paris green and the importance
of
" sticky
banding " "vras called attention to. Information concerning this pest is given
in
leaflet Xo. 4
; but it
should be pointed out that the use
<jf quassia
or
soft-soap wash
is
now known to be quite useless
for these biting-mouthed insects, and
also that when Paris green is used animals may V)e kept under the trees. It may also be pointed out that
JVyssia zonaria has no importance
as a fruit pest, feeding only on yarrow and dog-rose, nor is Phygalia
Ijilosaria found on fruit-trees, its food plant being the oak. The Winter Moth was
also reported
as damaging the leaves of
apple and pear at Glazebrook, near Manchester. The same correspondent sent some flies belonging to the Bihionidce. They have no connection with the
caterpillars
as
supposed. These
dipterous insects belong to the species Bihio marci (St. Mark's Fly). The
larv?e
live
in the
soil and some's^diat resemble small " leather-
jackets "
; usually they
occur
in
masses, and seem
to do
no
little
harm to the roots
of grass and other plants. Great numbers of this
species and B. hm'tulamis have appeared this
year, and have been
sent by numerous other correspondents. {Diplosis jpjjrivora,
Ptiley.) Pear
fruitlets
sent
to
the
Board
from
Brackenwaite, Wigton,
Cumberland, were attacked by the Pear JMidge
{Diplosis jtyrivora). This
same
pest
was
reported
from
Glazebrook, near ^lanchester. It
is interesting to note the northern extension
of this
pest and
its
much
later period
of reaching the mature
larval stage in the north. Drenching the ground under the
trees with
paraffin emulsion either
when the larvte
are
falling or
as the
flies
are hatching out
is most
beneficial. An unobserved
feature
in
the
attack
of this pest may
here be mentioned. When the fruitlets are " struck by the fly," the^
swell much more rapidly than
the sound
ones, and can thus always
be told on the tree by their being often twice the
size
of a healthy
fruitlet. Scale
Disease
and
False
Scale
amongst
Fruit
Trees
and
Bushes
(Apple,
Pear,
Gooseberry,
Currant,
etc.)- Specimens sent to the Board of Agi'iculture from Hayling Island
of apple,
pear,
quince, gooseberry and
currant,
all
supposed
to be Reports
to the Board of Agriculture. 23 attacked by scale, were in some instances perfectly healthy
; in others,
scale insects (Coccida^) were causing disease. The attention
of growers
is drawn
to the normal appearance of
bark and rind of the different fruit
trees, as
it
is not an uncommon
thing to have
cj^uestions
asked regarding the spots present on the
twigs of various
fruit
trees and bushes, which are
often,
as
in the
present case, mistaken for scale insects. The
following
specimens
sent may
]3e
taken
as
examples
of
this
: A. (Apple
;
an
exceptionally bad
case
of Quarrenden
apple
fourth year.) This specimen was covered with the Apple-bark Louse or Mussel
Scale {Mytilaspis pomorum) (Fig. 3, a). B. (Pear, presumed
to be
scale
of a
different
variety, somewhat
like bark blisters.) This twig
was
quite clean
;
the small grey spots (Fig. 3, h)
are
normal bark
spots, but might
easily be mistaken
for the San Jose
Scale {Aspidiotus perniciosus), or the Japanese Fruit
Scale {Diaspis
amygdali). C. (Gooseberry with
scale. This
scale
in
most
cases has
not
been noticed to leave a protruded egg trail. Does it
? See E.) This specimen
contained two specimens
of
the Brown Currant
Scale {Lccanium
ribis) ; no white protruding egg mass,
i.e., no white
cottony nest is formed so as to protrude from the scale in this species,
as
in
the White Woolly
Currant
Scale {Pulvinaria
rihesii). The
young scales
of this
species
are much flatter than the old and paler
in colour. D. (Currant with remnants of " egg trail.") Although no
scale
is
attached
to
the
twig
of currant
sent
it
contained a large cottony mass of eggs and wool. This resembles the
cottony cushion seen in Pulvinaria rihesii (vide C). This scale can
easily be
told from
L. ribis by being raised up off
the rind by a pad of white wool. E. (Gooseberry with remnant of egg trail, etc.) This spray of gooseberry had two mature scales of Lecanium ribis
and several smaller ones
;
also a mass
of white wool with no scale
attached. The scales are the same as C. Possibly the white wool is
due to the same species as D. F. (Apple Branch, grey spots on bark.) F. Life-history of Scales (Coccidcc.) The eggs of the Coccidie
are found under the
scales
;
the young
scales are minute, active, six-legged insects with projecting antenme
(" horns ") and
often
hair-like
processes. They can only be
seen
with
a
strong
magnifying
power. Scales
(Coccidre)
are provided
with a long piercing mouth, which the
larvse thrust
into the bark,
leaf or fruit. The larvte then degenerate, and by degrees form over the
body a scaly covering—beneath which you
find the mature female,
which
is legless
; the male scale insect is rare, and imlike the female
is active, having a pair of wings. Scale insects on trees in the open
pass the ^vinter in Great Britain mostly in the egg stage beneath the
scales, a few as immature females. Reports
to the Board of Agriculture. 25 These small red
spots
are not
scales nor the stage
(jf any insect,
the qnince having red bark spots. It
is important
to notice whether we have
scale on the tree or
whether the markings are normal plant structures. Scales are often
very harmfid in this country and should be checked before they get
the upper liand of the tree and sap its vitality. Three washes may be used for scale insects
: 1. Paraffin emulsion. 2. Eesin wash. 3. Caustic alkali wash. The
first
(paraffin emulsion)
is
chiefly
of use when the young
scale
insects
are
emerging
from
beneath
the
scales
;
observations
personally by the grov/er must be made, as the times of hatching vary
very much. Caustic alkali wash
is the most
beneficial
to
use (^•ide article in
the September number of the Board " Journal"). Fumigation with hydrocyanic
acid gas
is the best scale remedy,
but is difficult to apply to large trees. Should this be employed, full
information will always be sent from this Department. Winter washing with caustic alkali wash
is strongly advised. Scale
Disease
and
False
Scale
amongst
Fruit
Trees
and
Bushes
(Apple,
Pear,
Gooseberry,
Currant,
etc.)- (Apple Branch, grey spots on bark.) The grey
barnacle-like
spots
referred
to
are
not
scales
or any
insect, but bark spots. The grey
barnacle-like
spots
referred
to
are
not
scales
or any
insect, but bark spots. G. (Quince, red spots on bark.) G. (Quince, red spots on bark.) First Report on Economic Zoology. 24 Fig. 3. scale, false scale and effect op fbost and canker. <i, Mussel scale on apple wood
;
.v, the scales
;
(il, mussel scale enlarged
; a2, its eggs
;
6, pear
twig showing grey scale-like spots (normal
plant structure)
;
c, apple twig blistered
!iy host and canker (?), not insect work
; u and
c, blistered area. Fig. 3. scale, false scale and effect op fbost and canker. <i, Mussel scale on apple wood
;
.v, the scales
;
(il, mussel scale enlarged
; a2, its eggs
;
6, pear
twig showing grey scale-like spots (normal
plant structure)
;
c, apple twig blistered
!iy host and canker (?), not insect work
; u and
c, blistered area. Reports
to the Board of Agriculture. The Apple Bark Louse or Mussel Scale.
(Mi/tilasjns pomorum, Linn.) Several correspondents of the Board of Agricultm^e have reported
damages
to apple
trees by the mussel
scale, namely, from Eomsey,
Tarporley, Hayling
Island, and other places previously mentioned. One
con-espondent
thought
the
scale
connected
with
thrips
and
canker fungus. The
best way
to destroy
this
scale
is
to use the
caustic
alkali
wash
in
winter, and
spray
in the
spring and
early summer with
paraffin emulsion. A full account of this pest is given on page 75. A full account of this pest is given on page 75. Spkayers. For bush-fruit
the
best sprayers
are the "Knapsack
sprayers,"
the " Eclair," the " Notus," the " Anti-pest," and others. Ordinary
syringes
are
of
no
use
in
washing
plants. The
insecticide must be sent out in the finest possible spray. Paraffin emulsion should be used
first in April
for currant scale
and
repeatedly
every
two
weeks
for
at
least
two
months
;
for
mussel scale in May, and likewise onwards. It is advisable to try caustic
alkali wash alone for the first year. This need not be put on by a " mistifier," as the object is to satm-ate
the tree, not to spread a fine even layer of the wash over it as when
Paris screen or emulsions are used. Scale Insects on Plum Trees. Plum
trees
sometimes
sufter
from
scales. The
Oyster-Shell
Bark Louse (Asjoidiotus
ostrecqformis, Cmtis)
is found on plum. A
brown Lecanium has also been found, but was not identified. The Japanese Fruit Scale {Diaspis amygdali) also occurs on plum,
and thus care should be taken
to examine all Japanese cherries that Plum
trees
sometimes
sufter
from
scales. The
Oyster-Shell
Bark Louse (Asjoidiotus
ostrecqformis, Cmtis)
is found on plum. A
brown Lecanium has also been found, but was not identified. The Japanese Fruit Scale {Diaspis amygdali) also occurs on plum,
and thus care should be taken
to examine all Japanese cherries that First Report on Economic Zoology. 26 may be bought. Nuinbers of these fruit trees are introduced and the
scales may be
also
l)rought over with
the
plants. If the
scale
is
seen the plants should be fumigated or destroyed. The Brown Currant Scale {Lecanium
rihis) has
also been found
on
plum
trees,
especially
in
gardens when
the
trees
are
grown
against walls. Eggs on Apple Trees and a further Remedy
for Mussel Scale. (i) Apple
shoots
sent
to
the Board
of Agiiculture from South
Norwood were covered with the eggs
of (1) the Eed Spider {Bryobia
pruni),
(2) the Apple
Sucker {Psylla
mali). The
former are
red
globular eggs, the latter are elongated
o^'al and white. Two only of
the Psylla eggs could be detected. The majority of the Bryobia eggs Reports
to the Boai'd of Agriculture. Tj were shrivelled and killed, as also were the two Psylla eggs
; some of
the former, however, had liatched
out. Psylla mali has been most
destructive
during
the
past
season,
both
in
Worcestershire
and
Herefordshire, and has been more or less troublesome in Kent. For
these two pests a combined wash of paraffin emulsion and
sulphur should be used. Ordinary paraffin emulsion may be made
by mixing
e(iual portions
of boiling
soft soap
solution and paraffin
together, and then churning them up by means of a force pump until
a creamy emulsion is produced. When required for use, this may be
mixed with twenty-five times its bulk of warm water. To every three gallons
of this
dilute emulsion, add one ounce of
liver of sulphur and well mix. Spray in a fine spray, so that every
part of the tree is wetted. (ii) The only thing to do
after the buds have
burst
to destroy
mussel
scale
is
to wash with paraffin emulsion two
or three times
during
the late spring
and
early
summer. Commence
at
end of
April—if possible a look out should be kept— and
as soon
as any
signs
of the yoimg active
scale
insects
are noticed crawling
about
the
trees washing
should
be
carried
out
;
the
corroding
effect
of
paraffin emulsion
is not
great on
the mature
scales,
but
it
soon
destroys the immature forms. The wash recommended
for the two former insects
will do also
for this scale. A General Wash for Fruit Trees. A general insect wasli required by a Gloucestershire correspondent
of the Board of Agriculture may be made as follo^vs
: A. (1) Dissolve 1 oz. of arsenate of soda in soft water and add to
16 gallons of soft water. (2) Then dissolve 3 ozs. of acetate of lead in soft water and add to
above and stir well
( = Arsenate of lead
vjash). (2) Then dissolve 3 ozs. of acetate of lead in soft water and add to
above and stir well
( = Arsenate of lead
vjash). B. Dissolve
1 quart
of
soft
soap
in
2
quarts
of
boiling
soft
water. Then remove from
tlie
fire and while
still boiling hot add
1 pint of paraffin and churn the whole together for ten minutes with
a
small hand
syringe. (For
use
alone
dilute
with
ten
times
its
volume of soft water (= Paraffin
emulsion.) For mixing with A,
add
about
two
pints
of
tlie concentrated
emulsion (B) and mix well. The combined wash
will then destroy
both biting or mandibulate and sucking or haustellate insects. By far the most successful wash
for " Apple Sucker "
is Quassia
wash as used in the Kent hop gardens. Aphides
{A.
mali, Fabr.) on Apple Trees. Some insects sent by a correspondent of the Board from Chesham
proved to be the Apple Aphis {Aphis mali), which has been doing an
enormous
amount
of damage
this
season (1902)
in
the
South of
England—many orchards being covered by them. Steps should be
taken
as soon as the plant
lice
are seen
to destroy them. This can
easily be done if the leaves are not too far curled up, but even then
some good can be done by spraying. The best wash to advise for this Aphis is soft soap and quassia, but
if the operator has proper sprayers, paraffin emulsion. It should be
pointed out that the wash must go on the under surface of the leaves. Two washings, at
a few days' interval, are usually necessary for this
pest. This
pest was
also
reported
as
doing
considerable damage
at
Tunbridge Wells, and also from Glazebrook, near Manchester. First Repoi't on Economic Zoology. 28 The Peach Aphis. Amongst
a number
of
Apliides
sent
by a correspondent from
Tunbridge Wells were specimens of the Peach Aphis (A2)his amygdali). The Peach Aphis
also
attacks
nectarines and causes the
leaves
to
curl up and
to
fall,
often leaving the branches quite
bare. It also
feeds on
the
sloe and tobacco
plant. Another
species, Hyaloptei-^s
pruni,
Fabr,,
also attacks the peach, but from the remains
sent the
species
in
question
here
is
A. amygdali. These
Aphides
have
nothing
to
do
with the
curled
fleshy leaves
seen on
peach
trees
caused by the fungus Exogcsius deformans. It
is
advisable
to
use
quassia wash only on peach,
as
paraffin
emulsion might damage
the foliage and
the young
fruit, the peach
being much tenderer than the apple or plum. Canker Fungus {Nectria
ditissima) on Apple Twigs
mistaken for Insect Work. The
effect of " canker " is often mistaken for insect work. Apple
twigs Avere sent
to the Board of Agriculture from Bournemouth with
brown
blister-like
patches
over them
; these were
attacked
Ijy the
canker
fungus
Nectria
ditissima
(fig. 3). The
shoots
had
been
probably damaged by frost early in the season
; canker becomes more
prominent lower down the tree. Another correspondent from Eugby
also
sent
twigs with the
typical canker of apple
;
the small
scarlet
fruit
or perithecia of the fungus were present on most
of the twigs
sent. There
was
a
lot
of
this
disease
showing
these
blister-like
patches this year (1902). Further Information
re Winter Washing of
Fruit Trees. In answer to an enquiry of a correspondent of the Board of Agi-i-
<3ulture,
re winter washing
of fruit
trees,
it was
pointed
out
that
" washing "
and
" spraying "
of
fruit
and
other
trees
are
mei-ely Reports
to the Board of Agriculture. 29 different terms for the same treatment. Hop growers call the process
" washing,"
fruit growers
both
" washing " and
" spraying."
In
all
insecticides
it
is well
to put
the wash on
the foliage
or fruit
in
as
fine a mist as possible, but with the Caustic Alkali wash this
is not
essential. The wash may be syringed over the trees
thickly, but
it
is advisable to use a proper sprayer or washer such as the Strawson
" Anti-pest."
After spraying with this wash the machine should be
well cleaned out with cold water. No
care
is necessary regarding
the buds as long as they have not commenced to burst. Three quarters of a pound of soft soap for the ten gallons of wash
may be used in place of the treacle previously advised. India rubber
gloves are sometimes worn by the sprayer, but are not necessary. It
is far better to fix a circular disc pointing downwards on the spraying
tube so that any wash that runs down will fall clear of the hands. Another Board correspondent was informed that ordinary treacle
may be substituted for coarse agricultural treacle in this wash. Certain gardening papers have
raised an objection
to the use of
treacle
or
soft
soap
in
this
wash,
but
it
is
certainly
a
beneficial
ingredient. Land Bugs on Chrysanthemums.
{Lygus iiratensis, Eabr.) The
insects sent
to the Board
of Agriculture by a correspondent
from South Norwood,
S.E., are
Hemiptera-Heteroptera
(Bugs) and
belong
to the
species known
as Lygus ijvatensis, the L. canvpestris of
Linnseus. This
is a very common and widely
distributed
British
species and is sometimes harmful to various garden plants. There is,
however, no record of their attacking chrysanthemums. Several other
species of land bugs are injurious to garden plants,
including
the
so-called
potato
bugs,
Phytocoris
'pabulinus,
L. and
Lygtis containincdus, Fallen. These bugs injure the plants by sucking out the juices, puncturing
stem, leaf and blossom. The life-history of Lygus pmtensis is not known, but
it may be
mentioned
that the eggs are usually laid on the
plants upon which
the insects feed—these eggs give
rise
to the
larval
or louse stage
a
creature much like
the
adult,
but wingless
;
the next
stage,
the
pupal
stage, differs in having two bud-like processes on each side of
the body, the wing buds. These
plant bugs are injurious in all
three stages. Some winter
as eggs, others hibernate amongst rubbish in hedgerows, etc. Enquiry as to Poison for Moles. Several enquiries have been made
regarding
Moles. One
cor-
respondent wrote asking the best way to poison these animals. This Department knows
of no poison
of any use
in destroying
moles, but probably bisulphide
of carbon would be
as successful in
killing these animals as it is in destroying the Canadian Pouched Eat First Report on Economic Zoology. 30 or Gopher. This
Latter animal
is most harmful in America, but the
mole is not with us and its destruction should not be advocated. If
they are very numerous, as on the land of the BoaixVs correspondent
•they should be trapped alive and spread over the country. Section
II. Animals Injurious to Horticulture. Reports
to the Board of Agriculture. 31 In
a
similar
attack
in
hops by
a
species known
as
Calocoris
fidvomaculatus of De Geer jarring over boards
cleared the
invaded
gardens. The only washes found of any use are soft soap washes, especially
paraffin emulsion with an extra 3 lb. of soft soap to the 100 gallons. To be of much service
the wash must be used when the
insects
are
in
the larval
or pupal stage. A look-out should be kept in the
garden
to see where
this chrysanthemum pest passes the winter
;
if
the winter quarters are discovered steps should be taken
to destroy
the
insect
before spring, if
it hibernates in the adult
stage. If the
species passes the winter in the egg stage on the chrysanthemums,
the plants should be sprayed with
paraffin emulsion
as soon as the
young larvie are seen to appear. Treatment. The
only
remedies
of any
avail
against
these
creatures
are
(i)
collecting them by jarring the plants over taiTed boards held on
each side and (ii) treatment by washing. Reports
to the Board of Agriculture. The Destruction of Ants. The
following information was
sent
to
a con-espondent
of
the
Board of Agriculture at Kingston-on-Thames concerning the method
of destroying ants damaging grass and clover. First find out the
ants'
nest. This may be under the ground, in
Avhich
case
it can only be detected
by following
the ants and
so
finding the opening in the soil down which they descend
;
nests may
also be formed under dome-shaped masses of earth, etc., above ground
or they may be
under stones and
rotting
wood, according
to
the
species of ant concerned. Having located
the
nests, make
a hole in each one about
eight
inches
in
depth
and
then
pour
into each hole from
1
to
2
ozs. of bisulphide of carbon, according
to the
size
of the
nest, and
fill
up
the hole
at once with
earth. If the ground
is regularly under-
mined with
ants' nests
and
tunnels, treat the
soil with the same,
making holes every three feet apart, and pour into each hole 2 ozs. of
the bisulphide of carbon. This is as a rule not necessary for ants, the
nests only needing treatment. Evening
is the
best time
to attack
them. Care must be taken not
to put a light near the Ijisulphide of
carbon or to
let
it come
in contact with hot metal, as
it
is highly
inflammable. This
is the only plan by which ants can
l)e
cleared
out of the soil and has so far always met with success. There
are
veiy
few
parasitic
enemies
of
ants. A
genus
of
Ichneumon flies, Masmosoma, is one of the few parasites
that attack
them, whilst a brilliant Chalcid, Eucharis myrmecim, is known to prey
on the
large Australian Myrmccia. A small
fly, Pliora formicarum,
Verrall, lays
its
eggs on
ants,
inside which
the
larvai
live. Ants First Report on Economic Zoology. 32 are
also attacked by
mites. Numerous
biixls, of course, prey upon
them. The Destruction of Subterranean Insects and
other Ground Garden Pests. The Destruction of Subterranean Insects and
other Ground Garden Pests. A box containing the foUoM^ing
creatures was
sent by a corres-
pondent
of the Board of Agriculture from Glasgow, with a request
for information as to how to clear them out of his garden. i. Wire-worm—The larval
stage
of the click beetles {Elaieridce). These are hard, shiny and bright yellow. ii. Julida3, including ii. Julida3, including a. Two species of Jul us. h. A species of Polydcsmv.s. h. A species of Polydcsmv.s. iii. Scolopcndridcc, including GeojMlus
loiigicornis. The wire-worms are, of course, very injurious, and so also
are the
Julidcv. A small white Julus
sent was an immature
form,
but
a
different
species
to the large dark snake millepede {Jidus terrestris,
Linn.). The Polydesmus can easily be told by the sides of the body being
notched and by its more or less flattened form. It was
too damaged
to
identify,
l)ut was probably Pohjdesmus comiilanahis, Linn. It
is
also injurious to plant life. Geopliilus loiigicornis, the long snake-like
yellow species with one
pair of legs
to
each
segment, is a centipede
and
is
beneficial, probably
feeding
upon
the young
Julid;e. The
pests may
1 )e destroyed in the following way
: (ii) By trapping, (i) By fumigation. Directions
for the Employment of the Gas
Treatment under Glass. Several
enquiries
ha\'e
been received concerning the destruction
of greenhouse pests. Various methods of fumigating plants under glass
are employed,
such as sulphur fumes and tobacco smoke. Neither of these are
as
good as
the employment
of
hydrocyanic
acid
gas. For
Aphides,
Ked
Spider, and Wood Lice the following quantities are necessary
2
ozs. of cyanide, 4
ozs. sulphuric
acid,
7
ozs. of water per 1000
cubic
feet
of space. Proceed as follows
: Pour the water into a jar,
then add the acid to the water. Wrap up
the cyanide in a piece of
blotting-paper and drop it into the jar of acid and water from outside
the house. This can be done by tying the cyanide bag on to a stick
with a longish piece of string, then close the door or window. Leave
the house shut up for at least three-quarters of an horn-, then open all
doors and windows
to
ventilate
freely
; be
careful not
to
enter the
house
for
at
least an hour
after the doors and windows have been
thrown open. Fumigate
at dusk and when the foliage
is dry. The
temperature
must not be above
G0°
F. The
best
temperature
is
50^ F. ; above 60^ F. the foliage gets
scorched, as
it
also does
if you
fumigate
in bright
light. If the house
is more than
10,000
cubic
feet two jars would be necessary, but up to that size one is ample. The
result of one experiment may be quoted here. Greenhouse,
2,000 cubic
feet, containing Chrysanthemums in full bloom, severely
infested with Green Fly. Cyanide 3^
ozs., acid
5
ozs., water 9
ozs. Temperature 50° F. Time 25 minutes. 1 hour after sunset. Various methods of fumigating plants under glass
are employed,
such as sulphur fumes and tobacco smoke. Neither of these are
as
good as
the employment
of
hydrocyanic
acid
gas. For
Aphides,
Ked
Spider, and Wood Lice the following quantities are necessary
2
ozs. of cyanide, 4
ozs. sulphuric
acid,
7
ozs. of water per 1000
cubic
feet
of space. Proceed as follows
: Pour the water into a jar,
then add the acid to the water. Wrap up
the cyanide in a piece of
blotting-paper and drop it into the jar of acid and water from outside
the house. Teeatmext. Peat moss manure
is always attractive
to Myriapoda and other
pests and should be
avoided. Lime
if applied
in proper quantity
always seems
to check the
increase of millepedes, but has no
effect
on the wire-worm. Both wireworms and millepedes are prominent
garden pests and can only be treated in two ways, viz
: (i) Fumigation
for subterranean
insects and other animal pests
is best carried out by using bisulphide of carbon. Proceed as follows
:
Make a small hole in the flower bed
or border every two yards and
pour in h oz. of the bisulphide
of carbon and
close up each hole as
soon
as
the carbon
is poured
in. This must be done
so
that the
bisulphide
of carbon does
not touch
the
roots of a
plant, thatis, it Reports
to the Board of Agriculture. 33 must be put in the earth between the
pLants. Care must be
taken,
as it is of an inflammable nature and the fumes are also poisonous. (ii) Trapping largely employed for wire-worm consists of placing
pieces of carrot, mangel, beetroot or turnips in the ground, and taking
them up
every few
days
and
collecting
the
wire-worm
that
are
feeding upon
the
"bait."
Millepedes may
also
be
caught
in
this
way, but for them
large
hollow, more or
less
rotten, roots form the
best trap. It has also been found that the small millepedes {Jidus
pulcliellus) may
be
caught in numbers
by
placing cabbage
leaves
soaked
in
a
solution
(1
oz. to
the gallon
of water)
of
Paris green
on the ground. The millepedes come
to
the
surface
at night and
feed upon the leaf and are so poisoned. Bisulphide
of carbon treatment
is
best
;
failing
that, some good
may be done by trapping by means of ground bait. White Grubs or Maggots
{Phorhia
hrassicce,
Bouclie)
causing
great
damage
amongst
Cabbages,
Carrots and
Broccoli. The larvae and puparia of the Cabbage Eoot Fly {Phorhia hrassicce)
were reported
to the Board
of Agriculture from
Castle
Croft, near
Wolverhampton,
as doing considerable harm. Several
other corre-
spondents
reported
the same
pest. The
different
reports
sent out
are here
united. This fly
is
a great
pest
in most cabbage-growing
districts
in
Great
Britain, and
also
causes endless harm
in North
America. The only publication of value on this pest is by Professor Slinger-
land, of Cornell University Agricultural Experiment Station. (" The
Cabbage Eoot Maggot,
etc.,"
Bull. 78, Cornell Univ. Exp. Station,
Nov., 1894.) The
flies, which
are
very
like
the
house-fly,
appear
all
the
summer
in successive broods. Maggots may be
found
as
late
as
November. These latter pupate in the
soil, but apparently some of
the
adults
also hibernate and come out and lay
their eggs
in the
spring. Generally
there
are
three
broods
in
Great
Britain, and
undoubtedly the majority
pass the winter
in
the
puparium
stage
either in the ground or in the heaps of cabbage stumps and roots one
sees so frequently on the farm. Directions
for the Employment of the Gas
Treatment under Glass. This can be done by tying the cyanide bag on to a stick
with a longish piece of string, then close the door or window. Leave
the house shut up for at least three-quarters of an horn-, then open all
doors and windows
to
ventilate
freely
; be
careful not
to
enter the
house
for
at
least an hour
after the doors and windows have been
thrown open. Fumigate
at dusk and when the foliage
is dry. The
temperature
must not be above
G0°
F. The
best
temperature
is
50^ F. ; above 60^ F. the foliage gets
scorched, as
it
also does
if you
fumigate
in bright
light. If the house
is more than
10,000
cubic
feet two jars would be necessary, but up to that size one is ample. The
result of one experiment may be quoted here. Greenhouse,
2,000 cubic
feet, containing Chrysanthemums in full bloom, severely
infested with Green Fly. Cyanide 3^
ozs., acid
5
ozs., water 9
ozs. Temperature 50° F. Time 25 minutes. 1 hour after sunset. D First Report on Economic Zoology. 34 Eesult. Every Apliis
killed, also
slugs,
flies, wasps and
butter-
flies. Xot a petal
or
leaf injured. Cost
5fZ. Journal
S. E. Agric. College. The quantities given first are, however, now found most successful. Prevention and Teeatment. The results obtained from a long series of experiments conducted
by Professor Slingerland seem to show that
only two things can be
done
to mitigate the
evil caused by the Eoot Maggot. As
a pre-
ventive
the
only
effective
device
is
to apply around
each
plant,
when it is set out, a disc made of tarred paper or card. These can be
cut out by machinery in large numbers, and as placing them around
each
plant
before
it
is
set takes
so
little time the plan has been
adopted on a large scale by many American growers. A plan of the card disc invented by Mr. Goff is given on page 35, A plan of the card disc invented by Mr. Goff is given on page 35,
This
disc (A) must lie flat on the
surface of the
soil
to
stop the
flies from crawling beneath. Reports
to the Board of Agriailtttre . Willow Beetle at Norwich.
{Sarperda carcharias,
Linn.) ' A beetle received by the Board from a correspondent at Thorpe,
Norwich, proved
to
l^e
one
of
the
longicorn
beetles—the
largest
British
species
Sarperda carcharias,
Linn. It
is found
chiefly in
the
Fen
districts, and was
at
one
time
very common
there, but
appears to be now more local. It is found in and about old willows. The
larva? burrow into
the wood of willow, aspen, and poplar
; they
do considerable damage to a tree, but are seldom sufficiently abundant
to call for any remedial measures. Reports
to the Board of Agriailtttre . 1
qt. soap dissolved in
1 gal. of boiling water, and
1 pt. of crude carbolic mixed with the abo\'e. Pour round the roots of each plant. Reports
to the Board of Agriailtttre . 35 The only other plan found
to be
successful
is
the use of bisul-
phide of carbon or carbolic acid, the former injected
into the ground. This
treatment
on
a large
scale would
be
costly
in
Great
Britain
and need not be referred to here. Gas-lime has been found
of benefit, but to be so it has to be put
on strong and may damage the next
year's crop
to some extent
;
at
other times
it
is a complete failure. Gas-lime to be
of use must be
put on so strong that the land often requires a year's rest. All cabbage stumps and other
roots
after an attack
should
be
burnt. A dressing of superphosphate
of lime has been found beneficial
on the Continent. Fig. 4. stem of plant passing through card disc
lying close on the ground. A, Star-shaped
slit so as to allow stem
to pass through card
; B, Slit to push
stem up to A. Fig. 4. A, Star-shaped
slit so as to allow stem
to pass through card
; B, Slit to push
stem up to A. stem of plant passing through card disc
lying close on the ground. A, Star-shaped
slit so as to allow stem
to pass through card
; B, Slit to push
stem up to A. out has also met with success in preventing the fly from laying eggs,
but is by no means certain in action. By far the best plan is to give up growing cabbage for two years
on the land and plough deeply, or if hand
cultivation dig two
spits
deep. If it
is necessary to continue to grow cabbage again and again on
land, attention should be paid
to the method
of discing the plants
when being set. A small amount of soot and lime may also be put
in at dibbling time with the young plants. Carbolic acid in
soft soap and water
as follows was found
the
next
best remedy and preventative
after the
tarred discs
in a large
series of experiments in America. D
2 First Report on Economic Zoology 36 The mixture used was made as follows
:
1
lb. hard soap. 1
qt. soap dissolved in
1 gal. of boiling water, and
1 pt. of crude carbolic mixed with the abo\'e. Pour round the roots of each plant. The mixture used was made as follows
:
1
lb. hard soap. Section
III.
Animals Injurious to Forestry. Animals Injurious to Forestry. {Cladius
viniinalis.) {Cladius
viniinalis.) Some
larvai sent
to the Board of Agriculture from Brondesbury
attacking
poplars
were
those
of
the
Poplar
Saw-fly
{Cladius
viriiinalis). The eggs are laid on the leaf-stalk of the
poplar, which becomes
Swollen and bends over on each side so as to cover the eggs. The
young
larva
are green with
black
heads
;
at
the
second
moult they become orange and green with twelve large black marks
on each side, etc. When
full fed they are
entirely orange with the
black marks very prominent. The double cocoon is usually spun beneath loose bark or may be
between the leaves. They feed in companies and eat the epidermis
usually on the under side of the leaf. These is one brood which occurs in August and September. The
insect
is very common and
the
larvte sometimes do
some
harm to the leafage of young trees. They also occur on the willow and osier. Reports
to the Board of Agriculture. Reports
to the Board of Agriculture. 37 The trees should be cut down and burnt in the winter when the
larvse
and
pupte
are
safely
housed
in
the
wood. There
are
no
remedies
for the
Sirex
Flies, but
all damaged and diseased
timber
should be
cleared out
so
as
to check
theii-
increase. Damaged
or
unhealthy trees are mainly attacked. {SircM juvcnci's,
Linn, and
S. gigas,
Linn.) {SircM juvcnci's,
Linn, and
S. gigas,
Linn.) Fir trees in the neighbourhood of Bath have been reported to the
Board of Agriculture as being seriously attacked by the tM^o common
British Wood Wasps
[Sirex juvencus and Sircx gigafi),
the former
popularly called the
Steel Blue
Sirex,
the
latter
the
Giant
Sirex. The larvfe of these two insects, wliich burrow into the wood, remain
and pupate in the timber and may be found there during the Avinter. Infested
trees should be
cut down and burnt. They should not
be cut up into post and
rails, as is often done, as many of the
larviB
and puppe will hatch out after the wood has even been creosoted. Fir trees in the neighbourhood of Bath have been reported to the
Board of Agriculture as being seriously attacked by the tM^o common
British Wood Wasps
[Sirex juvencus and Sircx gigafi),
the former
popularly called the
Steel Blue
Sirex,
the
latter
the
Giant
Sirex. The larvfe of these two insects, wliich burrow into the wood, remain
and pupate in the timber and may be found there during the Avinter. Infested
trees should be
cut down and burnt. They should not
be cut up into post and
rails, as is often done, as many of the
larviB
and puppe will hatch out after the wood has even been creosoted. Sawfly Larvae on Willows. Some
Sawfly
larvae,
sent
by
a
correspondent
from
Heading,
belonging to the genus Nematus, were reported as attacking willows. A great number of Sawflies feed on the willow and osier, and several
of this genus Nematus have larva3 very similar
to the two
sent. It
was not possible
to say
for certain what the species was unless
the
perfect
insects were bred, but
it was probably the
species known as
Nematus conjugatus, Dbm. ; but
at the same time there were slight
differences seen
in the larv?e sent and the description of those of that
species given by Cameron. Another
species has
similar green and
orange larvse, N. croceus, Fall., but the black markings in those from
Eeading did not agree. Nematus pavidtis, Lep., does most harm
to
osiers in this country
;
but they were not that
species, as
the orange marks
are
larger, and
rthere were certain black dots which are not seen in N. pavidus. 38 First Report on Economic Zoology. Insect Galls on Osier Plants. Diseased osier-shoots were sent
to the Board of Agriculture by a
correspondent,
infested with
the
larvfe
of one
of the
Gall Midges
{Cecidomyidcc). The material sent was not sufficient to say definitely
the species of Cecid doing the damage, but probably it was Ceddomyia
salicina, Schrk. The following Cecid Willow
Galls, formed on
stems and twigs,
may occur in abundance in any part of Great Britain
: Those that form true leaf galls are not mentioned in this report. (1.) Cecidomyia heterdbia, Lw. The larvae live in the male flowers
and in rosettes on the leaves of salix, especially
aS'. amygdalina. These
larvae pupate in the galls. (2.)
C. salicina, Schrk. The
larvae
live
in the withered
tips of
the young
shoots
of salix—in
the
terminal
leaflets
of the
shoots
which wither away and form
a bud-shaped nidus. Three to eight
larvae inhabit each gall. (3.)
C. terminalis, Lw. The larvae
are
yellowisli-red and live in
bloated galls on the shoots of salix; 20-30 ova are deposited
at a
time. When the
larvae leave the shoots, many scars appear between
the healthy and galled parts. They pupate in the ground. (4.)
C. rosaria, Lw. The larvae form rose- shaped galls at the ends
of the boughs. They pupate in the rosette. (5.)
G. saliciperda,
Duf. Orange
larv£e
found, from
July
to
August, in the wood of young willows under
the bark, where they
form short irregular passages with gall-shaped
swellings, and cause
the bark to crack and become scabby. (5.)
G. saliciperda,
Duf. Orange
larv£e
found, from
July
to
August, in the wood of young willows under
the bark, where they
form short irregular passages with gall-shaped
swellings, and cause
the bark to crack and become scabby. (6.)
C. salicis, Schrk. The
larvae form
large woody galls on the
boughs, many in each gall, and pupate in the swellings they form. (6.)
C. salicis, Schrk. The
larvae form
large woody galls on the
boughs, many in each gall, and pupate in the swellings they form. (6.)
C. salicis, Schrk. The
larvae form
large woody galls on the
boughs, many in each gall, and pupate in the swellings they form. On making a careful examination
of the material
sent, one gall
was found to contain four orange-yellow Cecid larvae. Insect Galls on Osier Plants. Probably (as
most of the
galls were empty) they pupate in
the
earth, and thus
some good might be done by a heavy dressing of soot in the spring, or
sand sprinkled with
paraffin and
spread over
the
stocks or
stools. But until the
life-history is known
little can be done
to
eradicate
the pest. The Felted Beech Coccus.
(Cryptococcus fagi, Barensprung.) (Cryptococcus fagi, Barensprung.) Very few scale
insects are sufficiently abundant on forest trees in
this country to do much harm. One of the worst is the Felted Beech
Scale
(Ciyptococcus fayi)
of
Barensprung. This
insect
has
been Reports
to the Board of Agriadture. 39 reported from Castle Eden, Durham, by Mr. E. Burden, of the Castle,
and a request sent to
the Board of Agi'iculture for information. Mr. Burden, \mting
later
to me
at the British Museum, says
:
" I have
now noticed a large number of trees attacked more or less in the same
way. It looks like a regular epidemic, as it certainly has not appeared
in the same way for the
last few years. My forester
tells me that
they had the same
sort of epidemic on the Tyne, or in Northumber-
land, some fifty years ago (I think) and lost a lot of fine beeches."
It
has also been reported to the British Museum from Longwillow Hall,
Morpeth, from whence the following note was sent
:
" At a distance
the
tree looks as
if
it had been whitewashed
; when it is scraped off,
the yellow eggs or insects are
to be
seen. Two trees are covered on
the E. side of their stems. I remember a beech—not an old tree—in
Gloucester which was
affected in
the same
way, and
died
after
a
time. It smells something like the larva of a Goat Moth." This
scale
insect
chiefly attacks the trunk, but may ascend into
the boughs. The females give
rise
to
larvae in September, and they
envelop themselves
in
a white cottony
secretion, and then
cast
off'
their antennae and legs and remain for the rest of their lives devoid of
such appendages. The
adult female
is
a small orange-yellow
sac,
surrounded by a white mass
; these white patches
often unite and
form large felted masses, beneath which the larvae burrow and develop. These scale insects suck out the sap very greedily, and often do much
harm when present in large numbers. In time they cause the bark
to
peel
off the
tree and then decay and death may ensue. Large
numbers
of
trees are attacked in parts
of Surrey ;
it is also common
in Cheshire, Huntingdonshire, and probably occurs in small numbers
wherever the beech grows in Europe. The Felted Beech Coccus.
(Cryptococcus fagi, Barensprung.) The trees should be sprayed in the summer with strong
paraffin
emulsion twice at an interval of two days. In the winter they should
be sprayed with caustic alkali wash. The method of scrubbing the tree trunks
is too laborious
if the
attack is on a large
scale, and thorough spraying with warm paraf&n
emulsion is quite effective. Mr. Burbidge, of the Botanic Gardens, Dublin, has informed
Mr.. Newstead that
the
weeping
beech,
of which
there
are two kinds^
grafted on common beech stocks is not
affected by this coccus. The
stock may thus be attacked, but the weeping scion is not. This insect
is not
attacked by birds and
very rarely by
insect
parasites, according to Mr. Newstead. This insect
is not
attacked by birds and
very rarely by
insect
parasites, according to Mr. Newstead. Should the trees be cut down they should be burnt at once. First Report on Economic Zoology. 40 GROUP
F. GROUP
F. Animals
which
concern Man
as
being
injurious
to
his
worked-up Products of Art and Industry, such as (A)
his
various
Buildings and
larger Constructions, Habitations,
(B) Furniture and Books, Drapery and Clothing, (C) Food
and Stores."" Chermes
corticalis,
Kalt, on Pine Trees. Chermes
corticalis,
Kalt, on Pine Trees. Conifer shoots and bark, sent by a correspondent
of
tlie Board of
Agiicultiire from Holmleigb, Matfield, Paddock Wood, Kent, covered
with a plant louse which belonged to the genus Chermes, several
of
wliicli attack conifers. It is known as Chermes corticalis, Kalt. The apterous female is small, oval and yellowish-brown in colour,
the abdomen exuding a large quantity of white, flocculent silky matte
mixed with white meal
; beneath this the insect conceals itself. It is
rather firmly fixed to the bark by its short rostrum with long flexible
setae. The pupal
stage
is oblong and has transverse brown bars on
the abdomen,
the wing
cases,
antennae and
legs
also deep brown. These give
rise
to
the winged viviparous female, which has a black,
shiny thorax, and abdomen ringed and covered with masses
of white
wool
; the
legs
are
black, and
the fuscous wings have
coarse brown
veins. The
insects
are very harmful when present
in large numbers
;
the
trees should
. be sprayed with paraffin emulsion, especially when
the
larvae
are hatching
out, and the disease
in consequence rapidly
spreading. The
larvae were very active when the specimen arrived
in June. This
species
is
often
destructive
to
the
Scotch
pine
(Pinus
sylvestris) and Weymouth pine {Pinus strohvs), both on the twigs and
trunk. The white flocculent matter
is seen in June around the base
of the needles. There the wingless female produces her yellow eggs,
which gradually become brown. Numerous
natural
enemies
occur
in
this
species
;
the
ova
are
preyed
upon,
according
to Buckton, by
the
larvae
of Scymnus
dis-
xoideus, and by the Land Bug (Anthocoris fusca, Kalt), and by
the
larvae of Agromyza chermivora, Kalt. Reports
to the Board of AgricMlture, 41 GROUP
F. Animals
which
concern Man
as
being
injurious
to
his
worked-up Products of Art and Industry, such as (A)
his
various
Buildings and
larger Constructions, Habitations,
(B) Furniture and Books, Drapery and Clothing, (C) Food
and Stores."" I.
Furniture Beetles. Seveeal
enquiries
have been received from correspondents
of
the
Board concerning Furniture Beetles and Clothes Moths. The
so-called Furniture Beetles
are
usually known
as
" Death
Watches "
;
they are beetles belonging to
the genera Anohium and
Xestolmm. The group of beetles to which they belong, the AnoUina,
are mostly found
in old wood
; several are found in houses, the two
commonest being the Anobium
domesticum,
Foure, and Xestobivm
tessellatum,
F. Eleven
species
of AnoUina
are
found
in
Great
Britain
; the two
previously mentioned and A. paniceum,
L.,
and
ErnoUus
mollis,
L., are the domestic
forms. Furniture and wood-
work are chiefly damaged by A. domesticum and X tessellatum. The
species A. paniceum attacks
all kinds of stored goods, such
as flour,
bread, biscuits, medicinal
stores, skins, etc., and has been introduced
into most of our towns, but appears not to be common in Scotland. Anohium
domesticum
is a most
destructive
furniture pest, the
larvffi eating galleries
into the solid wood, and
often
so completely
riddling
it that
it
falls
to pieces
;
tables, chairs, and
furniture may
become so honey-combed by these pests that they suddenly collapse. The
exits
to
these galleries
are
seen
as
small round holes
in the
wood-work. In soft woods they soon cause complete destruction. Xesidbium
tessellatum
is also generally distributed, and occurs in
old
trees, such
as oaks and willows, but
especially in
old wood
in
churches and houses. It is also rare in Scotland. Both these species make a curious ticking
noise, which has given
rise
to their popular name of " Death Watch."
This noise
is made First Report on Economic Zoology. 42 chiefly during
the
pairing
season, and
is produced by
the
beetles
striking their heads upon the wood on which they are standing so as
to attract their mates, who make a similar noise in reply. This noise
is made during the day as well as at night, but is not so noticeable at
that time. Several species make
this ticking noise, but those most
often heard in houses
are A. domesticum and X tessellatum. It
is
said
that
the
larva
also
can
produce
this
sound,
but
it
is
not
definitely known to do so. The
larvse make quite long
galleries
into the wood,
and when
mature pupate in
little chambers from which
the
beetles escape by
eating their way out. II. Clothes Moths. Three
species
of
small
moths
attack
clothes, woollen
articles,
carpets, etc. These all belong
to the group Tineiim, and have now
become almost cosmopolitan
;
in origin they are probably Old World
species. The tliree species are the following
: i. The Case-making Clothes Moth {Tinea pellionella, L.). ii. The Webbing Clothes Moth (Tineola Usclliella, Hum.), iii. The Tapestry Moth {Tinea tapetzella, L.). They are all common and very destructive in this country, both in
private houses and in stores. 1. The Case-making Clothes Moth, Tinea jpellionella, L., is a small
moth with wing expanse
of nearly half an
inch, the
fore wings
are
yellowish-grey with three
indistinct brownish
spots, the hind wings
grey,
and
the
wing-fringes
grey. The
larvae
feed on
all
woollen
goods,
carpets,
furs, and
feathers. The moth appears in February,
and may continue in successive broods until November. In America
there appears to be only one brood in the north, the moths appearing
from June
to August, but in the south there are two or more broods,
the moths appearing from January to October. The eggs are very small and are usually laid on the food material. The larvse are small dull white
caterpillars, the head reddish-brown,
and the second segment with a dark brown plate behind. They form
a tubular, slightly flattish case in which they pass the whole of their
existence, the head and
first few segments and legs being protruded
when the caterpillars move from place
to
place. This
case has
fre-
quently to be enlarged as the larva grows
;
this is done by the insect
making a
slit half-way down the tube and then inserting a patch of
new material, and then
a similar
slit
is made and
filled in on
the
opposite side, and then the same
is done at the opposite half of the
case, the larva having previously turned round inside the tube
; when
the tube wants lengthening, additions are simply made at each end of
the tube. These larval cases are made from the material upon which
the insects are feeding, change of food thus changing the colour of the
case
; sometimes when feeding on variously coloured fabrics the cases
are thus multi-coloured. Inside each case
is lined by a
fine white
silk spun by the larva. Treatment. Where furniture
or woodwork
is attacked by these beetles and
their larvse, steps should be taken
at once
to destroy them, as they
return generation after generation to lay their eggs on the same object
until
it crumbles
right away. Amongst
the best ways
of
treating
attacked furniture are the following
: (a) Painting with a brush with corrosive sublimate
;
this
poison
kills the beetles as they make their exit. (b) Fumigating with hydrocyanic acid gas
; where small objects,
such as chairs, are invaded they may be put in some closed cupboard
and left in the fumes for some days. (This gas, one must remember,
is a deadly poison,
as well
as
the
cyanide
of potassium used in its
manufacture.) (c) Benzine may also be applied to polished
furniture, but
it
is
best used mixed with carbolic acid
; furniture so treated has, of course,
the polish taken off and will require repolishing. (d) Eooms in which these pests are present should be fumigated
every week during July, either with sulphur or hydrocyanic acid gas,
and then well washed down with
carbolic. Of course, during fumi-
gation all windows should have the
crevices, etc. papered up and the
doors
tightly
closed
If
hydrocyanic gas
is used
it must be used
with care, and should only be employed
in certain
cases
;
it could
not be used in high attics, as the windows should be opened from
the outside so
as
to allow the fumes to escape from the room before
anyone enters. In the case of attics, where windows cannot be opened
from the
outside, sulphur had
best be
used. In the hands
of an
expert a whole house may be treated with the gas. Reports
to the Board of Agriculttire. 43 II. Clothes Moths. When mature these " houses " are either spun
to tlie substance upon which the
larvse have been feeding,
or more
often the larvse wander to
the walls and
ceilings and then fasten the First Report on Economic Zoology. 44 tubes firmly to the surface with
silk. The pupal stage takes
place
inside the case and lasts from two to three weeks. 2. The Webbing Clothes Moth, the Tincola biselliella of Hummel,
is about
the same
size
as
the preceding
species, and has the front
wings pale ochreous, and more or less shining, without any spots
; the
hind wings
are
whitisli and
the
head
reddish-yellow. The brvn
feeds on a great
variety
of substances, such
as woollen
goods, furs,
feathers, the
linings
of chairs and
sofas, and has been found feeding
on cobwebs. The moth appears from March to October and produces
two broods in the year. The larva
is dirty white, and spins a silken
webbing as
it
progresses
over
its
food
material
;
no
true case
is
formed
as
in
the preceding
species
;
but when mature
it
spins
a
cocoon
of
pieces
of
hair or wool
of
irregular outline and
pupates
within
it. 2. The Webbing Clothes Moth, the Tincola biselliella of Hummel,
is about
the same
size
as
the preceding
species, and has the front
wings pale ochreous, and more or less shining, without any spots
; the
hind wings
are
whitisli and
the
head
reddish-yellow. The brvn
feeds on a great
variety
of substances, such
as woollen
goods, furs,
feathers, the
linings
of chairs and
sofas, and has been found feeding
on cobwebs. The moth appears from March to October and produces
two broods in the year. The larva
is dirty white, and spins a silken
webbing as
it
progresses
over
its
food
material
;
no
true case
is
formed
as
in
the preceding
species
;
but when mature
it
spins
a
cocoon
of
pieces
of
hair or wool
of
irregular outline and
pupates
within
it. 3. The Tapestry moth, the
Tinea
tapetzella
of
Linnaeus,
is
also
known as Trichophaga
tapetzella ;
its
fore wings
are black from the
base to the middle, then white clouded with grey towards the tip, the
hind wings are pale grey and the head
is white
; the wing expanse is
about three-fourths
of an
inch. It appears during June and July. II. Clothes Moths. The larva forms
galleries
in the
cloth
or other substance
it attacks,
these galleries being lined with silk. It affects carpets, horse cloths,
upholsterings, especially in carriages, also furs and skins. The pupal
stage takes
place
inside the
galleries. Heavy and coarse materials
are
attacked mainly by
this
species, which damages by
its
actual
burrowing into the material upon which it feeds. Treatment for Clothes Moths. Frequent removal, beating and shaking
of clothes,
etc.,
will do
much
to prevent the harm caused by these three
pests. Materials
which
are
liable
to be
attacked should
be put away in boxes and
cupboards with pieces of naphthalene in muslin bags placed here and
there. Exposure to sunlight and plenty of air in May and June will
do much
to prevent
clothes being spoiled by them. Benzine has a
very
deleterious
effect on
these
pests
and any valuable
materials
might be treated now and then with this substance. In the case
of cloth-covered furniture spraying with benzine
is
the most successful way of treatment. Large
dealers of carpets and
furs could always keep
their stock
free from attack by adopting cold storage. A temperature of 40° F. is protective. Reports
to the Board of Agriculture. 45 (Dermestes lardarius, L.) Some
insects
sent
by
Mr. Edgar
J. Lewis
to
the
Board
of
Agriculture,
and
which
had
been
attacking
and
causing
damage
to
winter-ciu'ed
bacon,
proved
to be the
Larder
beetle
{Dermestes
lardarius). This
insect
is common
to North America, Europe, and
Asia. It
attacks
not
only
bacon and
hams,
but
cheese,
horns,
skins,
feathers,
hair,
silk
and
other
dry
goods. Fresh
liams
and
bacon
are
not so
liable
to
be
attacked
as
those
that
are
slightly
tainted, improperly cured
or injured
in any way. The
beetles are
very
disposed
to lay
tlieir eggs
in any
crevice, and have probably
done so in this case where the muslin bags are sewn up. The larvie
are very minute when
first hatched and can
easily penetrate muslin
unless it is very fine. The larvaa as they mature bury themselves in
the bacon, but at first they feed on the exterior. Insects and Mites in Furniture. Some furniture and household pests
sent by a correspondent
of
the Board from Whitchui'ch, Glamorganshire, proved to be two species. They were (i) the
so-called " Death Watch " (Atropos divinatoria)—
the same name
is given to certain furniture beetles, Anohium tessel-
lation, etc.—and (ii) Mites belonging to
the genus
Glyciphagus, and
were
G. domesticus, De
Geer,
the
Glyciphagus cursor, Gerv. Speci-
mens have been sent
to A. Michael, Esq. for
identification. They
are both best destroyed by fumigation. Sulphur is usually employed,
but if both the pests are particularly abundant the rooms should be
fumigated
with
hydrocyanic
acid
gas
as
well. Eooms
should
be
well brushed down and the floors washed with
soft-soap and water. Books,
etc., which harbour the Atropos should be subjected
to the
fumes of benzine in closed boxes. Fumigation with sulphur answers
best for the
mites, but
is not
so
effective upon the Atropos, hence
hydrocyanic
gas
is mentioned. (A
full report on household mites
is given on page 120.) Teeatment. Bacon
is
best hung
as
is
sometimes done
in America,
in
thin
paper bags, care being talceu that
all
crevices
are
closed, or
else the First Report on Economic Zoology. 46 minute
larvie comiug from the eggs
laid on the paper may manage
to work their way through. When the larvae and
beetles
are found in the bacon the attacked
part should be cut away fairly deep, and well washed with a strong
solution of salicylate of soda or salicylic acid. After a bad attack the
store
room should be well white-washed
and
then fumigated with
hydrocyanic
acid
gas,
bisulphide
of
carbon
or
with
sulphur
to
destroy the beetles—the
first for preference, as it is safer to use than
the bisulphide and more effective than sulphur. The use of bisulphide
of carbon
to destroy the beetles and larvae
amongst
the hacon has been suggested and would be quite successful
if we had only to deal with those two stages and the pupal stage, but
unfortunately the
beetles
also
lay
their
eggs
in
and
around the
attacked parts, and
I have not
at present obtained any satisfactory
results of the action of bisulphide
of carbon or hydrocyanic acid gas
on insect and mite
ova. In
all
cases experimented with so far the
ova have not been harmed
to any appreciable extent. The infected
bacon or ham had thus
best have the attacked parts where many of
the eggs appear
to be
laid cleansed with
salicylic
acid. A second
fumigation fourteen days after the first is the safest plan to follow. Treatment. 1. Well
clean
out
the
barn
or
other
building
in
which
the
beetles
have
been
at
work
;
walls,
ceiling
and
floors
should
be
cleaned, washed with whitewash and soft soap and all refuse burnt. 1. Well
clean
out
the
barn
or
other
building
in
which
the
beetles
have
been
at
work
;
walls,
ceiling
and
floors
should
be
cleaned, washed with whitewash and soft soap and all refuse burnt. 2. Keep grain in bulk and constantly stir. 2. Keep grain in bulk and constantly stir. 3. Keep well ventilated with cold air and plenty of light. In a
warm climate ventilation would do no good, but cold air soon checks
their reproductive powers. 3. Keep well ventilated with cold air and plenty of light. In a
warm climate ventilation would do no good, but cold air soon checks
their reproductive powers. 4. If
the
store
house
or
barn
is
fairly
air-tight,
close up
all
openings where possible and then fumigate with bisulphide of carbon. Evaporate 1
lb. of the bisulphide
of carbon to every 1000 cubic feet
of space (about). Put the carbon about
the surface
of the grain in
flat saucers—the heavy fumes penetrate through the grain and
kill
all
forms
of
life, but
do
not harm
the
grain—leave
closed
for
twenty-four hours and then well ventilate and move the grain over. If the grain could be treated in closed bins so much the better
1
lb. of the bisulphide
to every 100
bushels
of grain
is
sufficient,
leaving for twenty-four hours. A caution must be given that this substance is
: A caution must be given that this substance is
: 1. Inflammable. 1. Inflammable. 2. Both the fumes and liquid poisons. A detailed report on this pest
is given in the Journal of the S. E. Agricultural College, No. 5, pp. 11-21, 1897. The infested grain given
to poultry would do no harm—the birds
would devour the insects as well. The infested grain given
to poultry would do no harm—the birds
would devour the insects as well. the Corn Weevil, the climate not being warm enough for C. oryzce to
flourish to any extent. the Corn Weevil, the climate not being warm enough for C. oryzce to
flourish to any extent. the Corn Weevil, the climate not being warm enough for C. oryzce to
flourish to any extent. {Calandra granaria, L.) {Calandra granaria, L.) Corn Weevils {Calandra granaria) were sent by a correspondent
of
the Board
of Agriculture which had been
attacking some
oats
stored
in a
barn. This
beetle, and a
closely related
one, the Eice
Weevil (C. oryzce, L.), which has also been forwarded by another cor-
respondent, from damaged Indian corn, are the most destructive corn
pests
in granaries,
stores,
ships
and
barns, that
are known. Not
only do they
attack
stored
corn, but
also
all
cereals
in
transit. Whole
cargoes
of
wheat, etc.,
are
often
destroyed
in
transit from
India, Australia,
etc. One
or two instances
are known
of
field attack near mills
in
Great Britain. The beetles lay their eggs in the corn, the maggots feed inside the
grain and there pupate. They breed
fairly rapidly in
this country
and may attack other stored goods than cereals. Reproduction may go on
all the year in
mills, but
chiefly takes
place in the spring and summer. The warmer the temperature the
more rapidly do they breed. In Great Britain we mainly suffer from Reports
to the Board of Ag7'icMltMre. 47 First Report on Economic Zoology., First Report on Economic Zoology., 48 Treatment. The bushes should be sprayed with cupmiii and Bordeaux mixture
some time before and after harvesting
; the first application should be
about two weeks before the rust spots usually appear. Bordeaux mixture may stain the fruit, so that for an early washing
before
the
fruit
is
picked cupram had
best be
used, and Bordeaux
mixture after harvesting. Neither should be used for three weeks before the fruit is gathered
as they are to a certain extent poisonous. BORDEAux Mixture. Dissolve
the
copper
in
10
gallons
of water, boil
the
lime and
treacle with a quart of water for half an hour. When dissolved, mix
them together and
stir them up
well. The mixture
is
then ready
for use. Fungoid Disease
in Black Currant Leaves.
{Septoria
rihis.) The currant leaves sent to the Board of Agriculture from Wickham
Market, Suffolk,
are invaded by
a fungus which produces so-called
Currant Eust or Leaf Spot. The fungus
is apparently Septoria rihis. This
disease attacks
all kinds
of currants,
and
appears,
as
a
rule,
about the beginning of July. It is
first noticeable
as
small brown
spots. Dull whitish
spots
also appear, but these may be due
to another fungus. Both may be
treated, however, as one, so far as remedies go. Pkeparation of Cltram. Measure
out \ pint
of strong ammonia (avoid
the
fumes), and
add it to 2 quarts of water. Weigh out 1 oz. of carbonate of copper,
wrap it up in a piece of copper gauze and suspend it by a copper wire
in the ammonia liquor. Let it remain all night. When required for
use dilute the blue fluid with 12 gallons
of water. This
is the
best
fungicide for all ripening fruit. BORDE
Copper sulphate
Lime
Treacle
AVater
,
Aux Mixture. 1
lb. lib. lib. 10 gallons. APPENDIX. Amongst other enquiries made
to
the Board
of Agriculture,
of
which short letters only were sent, may be mentioned the following
: I. Tapeworms in Sheep at Okehampton. A coiTCspondent
of
the
Board
sought
information
concerning
Cestodes in sheep. The writer was referred to an
article dealing with this subject in
the " Agricultural Gazette " for Jan. 20, 1902, p"40. The chief British
ovine tapeworm
is Moniezia
cvpcmsa, which
is very destructive
to
lambs in many parts of the country. Its life-history is not known. Treatment. All that cau
l)e done is to wash
as soon as the fruit
is gathered
with
Bordeaux
mixture
;
the
best wash
is
the
" Wye
Bordeaux
mixture," prepared as follows
: Copper sulphate (bluestone)
. . 1 lb. Lime
. . . . . . 1 lb. Agricultural treacle
. . . .1
llj. Water
. . . . . .10 ga Dissolve the bluestone in 10 gallons
of water, and
boil the lime
and treacle with a quart of water for half an hour. When the blue-
stone is all dissolved and the lime and treacle liquid fairly cool, pour
the
latter into
the bluestone liquid and stir well. It
is then ready
for use and will keep any time. The
soil should
lie well limed
in
the autumn, and
the
bushes
sprayed again early next year, about the
iii'st week in May. Reports
to the Board of Agriculture. 49 {Pucdnia pringsheimiana, Kleb.)
' {Pucdnia pringsheimiana, Kleb.) The gooseberries and leaves
sent are attacked by a fungus. This
fungus is the ascidium stage of Pucciiiia pringsheimiana of Klebahn. fungus is the ascidium stage of Pucciiiia pringsheimiana of Klebahn. The cluster cups or secidia occur on the gooseberry, both on the
leaf and fruit
;
the other stages affect certain species of Carex (sedges). It
is generally seen
in damp
places, but
is
rarely
in
sufficient
abundance to do any practical harm. As in tlie present case it is doing
considerable damage, any further notes will be gladly received. The cluster cups or secidia occur on the gooseberry, both on the
leaf and fruit
;
the other stages affect certain species of Carex (sedges). The cluster cups or secidia occur on the gooseberry, both on the
leaf and fruit
;
the other stages affect certain species of Carex (sedges). It
is generally seen
in damp
places, but
is
rarely
in
sufficient
abundance to do any practical harm. As in tlie present case it is doing It
is generally seen
in damp
places, but
is
rarely
in
sufficient
abundance to do any practical harm. As in tlie present case it is doing
considerable damage, any further notes will be gladly received. Reports
to the Board of Agriculture. 2. Black Wire-worm in Mangolds. A correspondent wrote
for information
concerning
Black
Wire-
worm attacking his mangolds. No specimen being sent and nothing being known of any creature
having
this popular name, no information could be given. Further
information on this subject will be gladly received. E E First Report on Economic Zoology. First Report on Economic Zoology. 50 LIST OF LEAFLETS PREPARED AND REVISED FOR THE
BOARD OF AGRICULTURE. Prepared. No. 68. Currant Aphides. No. 69. Tent CaterpillaTs. No. 70. Winter Washing of Fruit Trees. No. 71. The Colorado Beetle. Bevised and Enlarged. No. 1. The Currant Bud Mite. No. 2. Weevils. No. 12. The Gooseberry Saw
Fly. No. 14. The Raspberry Moth. No. 15. The Apple Blossom Weevil. No. 16. The Apple Sucker. No. 20. 1'he Magpie Moth. No. 22. The Diamond Back Moth. No. 28. Cockchafers. No. 30. The Codling Moth. No. 33. Surface Caterpillars. No. 34. The Woolly Aphis or American Blight. No. 35. The Celery Fly. No. 38. The Carrot Fly. No. 46. The Stem Eel-worm. No. 47. The Asparagus Beetle. No. 48. The Pea and other Thrips. No. 49. The Fruit Tree Beetle. No. 53. The Pear Midge. No. 62. The Pear and Cherry Saw Fly
No. 40. The Kestrel or Windhover. No. 42. The Short-eared Owl. No. 43. Titmice,
No. 44. The Common Lapwing or Peewit. No. 45. The Starling. No. 51. The White or Barn Owl. No. 54. The Spotted Flycatcher. No. 55. The Swallow. No. 6. The Field Vole. No. 57. The External Parasites of Poultry
No. 58. The Internal Parasites of Poultry. LIST OF LEAFLETS PREPARED AND REVISED FOR THE
BOARD OF AGRICULTURE. Prepared. No. 68. Currant Aphides. No. 69. Tent CaterpillaTs. No. 70. Winter Washing of Fruit Trees. No. 71. The Colorado Beetle. Bevised and Enlarged. No. 1. The Currant Bud Mite. No. 2. Weevils. No. 12. The Gooseberry Saw
Fly. No. 14. The Raspberry Moth. No. 15. The Apple Blossom Weevil. No. 16. The Apple Sucker. No. 20. 1'he Magpie Moth. No. 22. The Diamond Back Moth. No. 28. Cockchafers. No. 30. The Codling Moth. No. 33. Surface Caterpillars. No. 34. The Woolly Aphis or American Blight. No. 35. The Celery Fly. No. 38. The Carrot Fly. No. 46. The Stem Eel-worm. No. 47. The Asparagus Beetle. No. 48. The Pea and other Thrips. No. 49. The Fruit Tree Beetle. No. 53. The Pear Midge. No. 62. 2. Black Wire-worm in Mangolds. The Pear and Cherry Saw Fly
No. 40. The Kestrel or Windhover. No. 42. The Short-eared Owl. No. 43. Titmice,
No. 44. The Common Lapwing or Peewit. No. 45. The Starling. No. 51. The White or Barn Owl. No. 54. The Spotted Flycatcher. No. 55. The Swallow. No. 6. The Field Vole. No. 57. The External Parasites of Poultry
No. 58. The Internal Parasites of Poultry. Prepared. No. 46. The Stem Eel-worm. No. 47. The Asparagus Beetle. (A) BRITISH. Animals
bred
or domesticated by Man
for Food
or for the
use
of
their
products
in
industry
or
for
their
Services
as living things. Origin and Varieties of Domesticated Geese. Au
enquiry
as
to the
origin,
etc.,
of
Domesticated
Geese was
received from
the Hon. Florence Amherst. No
information
could
be
gathered
in regard
to Pomeranian Goose and
little concerning
the
Strasburg
Goose. The
latter is
a white goose with
blue eyes
and thus probably of Embden origin. There seem
to be
five
well-marked
varieties
of
Domesticated
Geese
:
(1)
Toulouse
;
(2) Embden
;
(3)
African
or
Indian
;
(4)
Brown China
; and (5) Wliite China. The main characters of these may be summed up as follows
: EEPOKTS
ON
ECONOMIC
ZOOLOGY
SENT IN
REPLY TO
VARIOUS
CORRESPONDENTS. E 2 53 Nech-fcathers curled or hoisted ; no knob to bill, 1. Toidouse.—Adult
grey
;
bill
reddish,
nail
white
or
flesh-
coloured. Eye brown or hazel, rim colour of bill. Abdominal pouch
well developed. Gosling greenish-yellow. 1. Toidouse.—Adult
grey
;
bill
reddish,
nail
white
or
flesh-
coloured. Eye brown or hazel, rim colour of bill. Abdominal pouch
well developed. Gosling greenish-yellow. 2. Embden or Bremen.—Adult white
;
bill yellow to orange
; nail
white. Eye
blue, rim orange. Abdominal pouch
small. Gosling
yellowish. Legs orange, claws white. 2. Embden or Bremen.—Adult white
;
bill yellow to orange
; nail
white. Eye
blue, rim orange. Abdominal pouch
small. Gosling
yellowish. Legs orange, claws white. First Report on Economic Zoology. 54 Bill knohbcd, note much harsher than in 1 and 2. Bill knohbcd, note much harsher than in 1 and 2. 3. African
or Indian.—Adult with back, wings and
tail
dark
grey
;
bill
and
knob
black. Eye
hazel
or
brown. Legs
orange
Dewlap under bill. 4. Brown CJiina.—Plumage much as
in
African
;
bill and knob
dark brown
to
black. Eye
hazel. Legs dark with
greenish tinge. Dewlap under bill. 4. Brown CJiina.—Plumage much as
in
African
;
bill and knob
dark brown
to
black. Eye
hazel. Legs dark with
greenish tinge. Dewlap under bill. 5. White
China.—Plumage
white;
knob
orange. Eye
blue. Bill orange with white nail. Legs orange with white claws. 5. White
China.—Plumage
white;
knob
orange. Eye
blue. Bill orange with white nail. Legs orange with white claws. There
is no doubt that the European Geese are descended from
the
C4rey Lag (Anser ferns), which has not only a wide European
distribution, but is also Asiatic. There
is no doubt that the European Geese are descended from
the
C4rey Lag (Anser ferns), which has not only a wide European
distribution, but is also Asiatic. The blue eye
of the Embden has a similar parallel in the Blue-
eyed White Chinese Goose. The blue eye
of the Embden has a similar parallel in the Blue-
eyed White Chinese Goose. The
presence
of
the
twisted
or
curled
neck
feathers
of
the
Toulouse, Eml)den and other Europeans is characteristic of the Grey
Lag, and both will revert much to the Grey Lag in appearance. The
presence
of
the
twisted
or
curled
neck
feathers
of
the
Toulouse, Eml)den and other Europeans is characteristic of the Grey
Lag, and both will revert much to the Grey Lag in appearance. GROUP
D. Animals which concern Man as causing bodily injury, some-
times death, to him, and in other cases disease, often of
a deadly character. Nech-fcathers curled or hoisted ; no knob to bill, The two Chinese
varieties
are evidently clearly descended from
the Anser cygnoides ;
the
blue-eyed
white
variety
having
been
obtained from the wild form by selection. Thus we get two parallel
cases
of
blue-eyed
white
varieties
produced from
different parent
stock. The African
or Indian Goose
is
due
to
crossing
between
the
Grey Lag and Chinese (White), Dipterous Larvce
in Hwnan Excreta. 55 Live Dipterous Larvae
in Human Excreta and
Notes on Species producing Myiasis. Mr. J. W. Bridge, of the University, Birmingham, sent two larvae
from
the
excreta
of
a woman
with
the
following
letter,
dated
Dec. 4th, 1901 :— I ask you
to
identify
the specimens which
I am
sending
here-
with. They were sent
to me by a doctor
at Shrewsbury who states that
they were passed per rectum by a patient
suffering from
cancer. If you
can give me any chie to their identification I shall be grateful. To this the following reply was sent
: The
larvae
you
send
from
the
excreta
of
the woman
suffering
from
cancer
are
those
of
one
of
the Anthomyiidte and
of
the
genus
Homalomyia. Certain species of these diptera are responsible for authentic
cases
of
internal
Myiasis. They
are taken
in with
vegetable
food and
retain their vitality and are sometimes passed in the fa3ces alive. They are
in no way connected with cancer. Most
of
the
cases
of human
Myiasis
are
due
to
the
larvae
of
Compsomyia,
CctllipJiora,
SarcopliUa,
Homalomyia,
Ochromyia,
Dermatohia and Auchmeroyia. Myiasis may be
either (i) cutaneous
or (ii) internal. Homalomyia
canicularis,
Linn.,
has
been
reported
by
Hagen
(Proc. Bost. Soc, N.H. xx. 107)
as
living in
the
larval
stage in the
urethra
of
a
patient. I'robably,
says
Nuttall,
a
case
of
pseudo-
parasitism. The following
other dipterous
larvae have been known to cause
external or cutaneous myiasis. The following
other dipterous
larvae have been known to cause
external or cutaneous myiasis. SarcopJiila magnifica, Schiner, which deposits
its eggs in wounds
on man and animals. SarcopJiila magnifica, Schiner, which deposits
its eggs in wounds
on man and animals. 56 First Report on Economic Zoology, Luciliu
eccsar, Linn., also lays
its eggs on wounds
; probably the
sheep-fly, Z. sericata, Meig., does the same. Luciliu
eccsar, Linn., also lays
its eggs on wounds
; probably the
sheep-fly, Z. sericata, Meig., does the same. The Senegal or Cayor Fly {Ochromyia anthropo'phaga, Blanchard). The
larva}
or Cayor worms develop beneath the skins of man and
animals and
often produce
serious Myiasis in Senegal. Dermatohia
noxialis,
Goudot,
also
lives beneath
the
skin
of man
as
well
as
animals,
occurring from
^Mexico
to
Brazil. It
is known
under
a
rariety of names, such
as the Macaw worm
in Cayenne, the Ura in
Brazil, the Torcel at Costa Eica, the Mozoquil worm in j\Iexico. Live Dipterous Larvae
in Human Excreta and
Notes on Species producing Myiasis. The
Maggot
Fly
of
Natal
{Anchmcroyia
{Bencjalia)
deprcssa,
Walker), also produces serious cutaneous Myiasis
;
this pest is one of
the Sarcophagidre. It
is not restricted
to Natal, but occurs further
up the coast, having been recorded from Delagoa Bay,
The range of
this serious cutaneous
parasite seems
to be limited
to the coast and
no further inland than 1000
feet elevation. It
is common from the
Tugela downwards (vide Agri. Journal, Natal Dept. Agri. and Mines,
vol. iv. p. 606, 1901. C. Fuller). Vide
also note on Screw Worm,
(p. 131). To Dr. Ernest George Annis, M.R.C.S., etc..
Medical Officer of Health, Greenwich.
Dear
Sir,—It is somewhat difficult to answer the questions in your
letter re the annoyance caused by
biting
insects
reported
to your Com-
mittee without seeing actual specimens of the pests. Correspondence on the Mosquito Annoyance at
Blackheath. The following
letter was received
on November the
6th,
1901^
from the Public Health Department, Borough of Greenwich
: Dear Sie,—Complaints have been made to my Committee respecting
the inconvenience caused to
tlie inhabitants
in
this neighbourhood by the-
bite or sting- of insects found in the neighbourhood of Greenwich Park and
Blackheath,
popularly supposed
to
be mosquitoes. I
shall esteem
it
a
favour if you can give me any information you may have respecting this-
class of insect found in that immediate neighbourhood. I am, etc., Ernest George Annis, M.P.C.S., etc., etc.,
Medical Officer of Hecdtli. To F. V. Theobald, Esq.,
Natural Histoi-y Museum. To F. V. Theobald, Esq.,
Natural Histoi-y Museum. Mosquito Annoyance at Blacklieath. 57 I know
of no records
of mosquitoes from either Greenwich Park or
Blackheath. Two
species
occur
in
abundance
in
some
of
the
docks
(London and Albert), namely, Culexjnpiens, L., and Cuhx (lorsaUs,M.Q\gQn,
and from the
latter dock I have received Culex pukritarsis, of Rondani,
Cidex dorsaUs
is a vicious
biter, and I believe occurs
all down the
river. Cidex pipiens
also bites severely at times,
the bites being followed by large
red oedematous patches. I am
afraid without
investigation on the spot I
cannot possibly help you further. I am, etc., Fred. V. Theobald. To F. V. Theobald, Esq.,
Natural Histoi-y Museum. To F. V. Theobald, Esq.,
Natural Histoi-y Museum. Mosquito Annoyance at Blacklieath. BoEOUGH OF Greenwich
Public Health Depabtment,
Town Hall, Greenwich Road, S.E,,
12th November, 1901. 12th November, 1901. Dear Sir,—Please accept my best thanks for the information you
have
so
kindly
supplied
to
me, and
I
will endeavour
to
obtain
a few
specimens
of the
insects referred
to and submit them
to you if you are
agreeable. I am, etc.,
E. G. Annis. E. G. Annis. British Museum (Nat. Hist.),
Cromwell Road, S.W.,
21st November, 1901. British Museum (Nat. Hist.),
Cromwell Road, S.W.,
21st November, 1901. Bbitish Museum (Nat. Hist.),
Cromwell Road, S.W.,
Wi A iil
1902 ,
Wi Apiil, 1902. To Councillor Walter Dannatt, Blackheath. To Councillor Walter Dannatt, Blackheath. Dear Sir,—Your letter regarding the above subject has reach<idme. In November of last year I had some correspondence from the Public Depart-
ment of Health of the Borough of Greenwich on this same subject. I wrote
to Dr. Annis, the Medical
Officer
of
Health, saying I could not give any
definite advice, as
I did not know what
the
biting
insects were. Much
depends on the species of Culex or Anopheles. Do you know, or can you give
me any idea of the character of the Culex. C. dorsalis and C. jJipiens occur
along the Thames. The former is a very vicious biter, and I have known
it
<3ause much annoyance in the docks and at Eochester. C. pipiens also bites
at
times, but may never
do
so
in some
districts. The two
Anopheles
breed in different ways. A. hifurcatus
is in the larval stage in the winter,
A. macuUpenms in the spring and summer, but the former occurs again in
the smnmer in larval stage. C. pipiens larvae occur from May onwards in
tubs, cisterns,
etc., mostly in June, July and September. Culex dorsalis
I have found in small artificial collections of water in July, but it probably
also occurs in May. I am afraid only a careful examination will help you. I
should
paraffin the ponds for Anopheles (1) in winter
;
(2) in May and
June
; for Cidex (1) May and (2) July and September
; much depends on
local circumstances. For instance, Anopheles larvte occur in ponds, ditches,
rivers and
canals, in stone troughs and
in
rain-water
baiTels. So much
ground has to be covered, but I fully expect the pests you have are Culex,
probably both C. dorsalis and C. pipiens, which will be much easier to get rid
of. I have some exact data
of
larval appearance at home, and I
will look
this up and send you. I expect you know all about the way of
treating
the water to kill the larva
;
if not, I shall be pleased to help you. Without knowing the district and local conditions I cannot help you as Dear Sir,—Your letter regarding the above subject has reach<idme. Bbitish Museum (Nat. Hist.),
Cromwell Road, S.W.,
Wi A iil
1902 In November of last year I had some correspondence from the Public Depart-
ment of Health of the Borough of Greenwich on this same subject. I wrote
to Dr. Annis, the Medical
Officer
of
Health, saying I could not give any
definite advice, as
I did not know what
the
biting
insects were. Much
depends on the species of Culex or Anopheles. Do you know, or can you give
me any idea of the character of the Culex. C. dorsalis and C. jJipiens occur
along the Thames. The former is a very vicious biter, and I have known
it
<3ause much annoyance in the docks and at Eochester. C. pipiens also bites
at
times, but may never
do
so
in some
districts. The two
Anopheles
breed in different ways. A. hifurcatus
is in the larval stage in the winter,
A. macuUpenms in the spring and summer, but the former occurs again in
the smnmer in larval stage. C. pipiens larvae occur from May onwards in
tubs, cisterns,
etc., mostly in June, July and September. Culex dorsalis
I have found in small artificial collections of water in July, but it probably
also occurs in May. I am afraid only a careful examination will help you. I
should
paraffin the ponds for Anopheles (1) in winter
;
(2) in May and
June
; for Cidex (1) May and (2) July and September
; much depends on
local circumstances. For instance, Anopheles larvte occur in ponds, ditches,
rivers and
canals, in stone troughs and
in
rain-water
baiTels. So much
ground has to be covered, but I fully expect the pests you have are Culex,
probably both C. dorsalis and C. pipiens, which will be much easier to get rid
of. I have some exact data
of
larval appearance at home, and I
will look
this up and send you. I expect you know all about the way of
treating
the water to kill the larva
;
if not, I shall be pleased to help you. Without knowing the district and local conditions I cannot help you as
I should wish
I think I said thi
t
the Offi
of Public Health Without knowing the district and local conditions I cannot help you as
I should wish. I think I said this to the Officer of Public Health. Without knowing the district and local conditions I cannot help you as
I should wish. British Museum (Nat. Hist.),
Cromwell Road, S.W.,
21st November, 1901. To Dr. E. G. Annls, M.K.C.S., etc.. Medical Officer of Health, Greenwich. Dear Sir,—I shall be pleased to examine the noxious flies that are
causing annoyance in your district whenever you care to send them here. Yours, etc.,
Fred. V. Theobald. Yours, etc., Yours, etc., Fred. V. Theobald. Borough of Greenwich
Public Health Department,
26th November, 1901. To F. V. Theobald, Esq.,
British Musemu (Nat. Hist.) To F. V. Theobald, Esq.,
British Musemu (Nat. Hist.) Dear Sir,—Yours of the 21st to hand, for which I thank you, and
I am endeavouring to obtain specimens of the insects referred to, but they
do not seem to be so prevalent in the colder weather. h I shall, however, be pleased to avail myself of your kind
offer when I
am in a position to do so. I am, etc., Yom'S,
E. G. Annls. I am, etc., Yom'S, I am, etc., Yom'S, E. G. Annls. First Report on Economic Zoology. 58 Further correspondence on
this subject was received from Coun-
cillor Walter Dannatt, of Blackheath, who, writing on April 1st, 1902,
states that
: We have been much troubled with gnats and
mosquitoes in this
neighbourhood, the last two summers especially, and I thought something
might be done to abate the nuisance. ... I proposed
to have a solution
of petroleum put in the ponds on Blackheath and the neighbourhood to
destroy the
larvae, and I thought
if
all householders who have rain-water
tanks and
tubs were
to put some
petroleum
into
thoir
receptacles
that
many of the Culex would then be destroyed. Will you kindly inform me
when the breeding season of
these insects is ? I may say that these
pests
have been so troublesome in this neighbourhood that most people dare not
sit in their gardens of an evening in the summer. Many people have been
laid up by bites from these insects. Bbitish Museum (Nat. Hist.),
Cromwell Road, S.W.,
Wi Apiil, 1902. Bbitish Museum (Nat. Hist.),
Cromwell Road, S.W.,
Wi A iil
1902 I think I said this to the Officer of Public Health. Any further information I can give you I shall be very pleased to do. I am, etc.,
Fred. V. Theobald. I am, etc., Fred. V. Theobald. Mosquito Annoyance at Blackheath. Mosquito Annoyance at Blackheath. 59 DONNINGTON, VaNBRUGH ROAD,
Blackheath, S.E.,
Wi April, 1902. To F. V. Theobald, Esq., M.A. Dear
Sir,—Many thanks for your kind and
interesting letter. I
am
Avriting
the London County Council on
the matter
of
treating the
ponds
at Blackheath. I
shall be greatly
obliged
if you
will favour me
with the modus operandi of using the petroleum, and
if any particular kind
is used, and where obtained. I presume
it will destroy any fish that are in
the pond ? There are some
Culices about now, but whether they bite
or what,
I
know not. My opinion
is, that the
species that troubles us most
in the
summer
are
rather
small,
and
I
have
noticed
them
settle
but
never
appear
to crawl
; whether that will prove what species they are
I do not
know. Thanks, I shall be very glad
of
the
data you
refer
to
of
larval
appearances. I suppose the enclosed large one
is a Cidex ;
there
are plenty in out-
houses now. My opinion
is, that the summer
species
is
smaller. I have
found a couple which were killed against the wall
last summer. Are they
Anopheles ? I am, etc., I am, etc., Walter Dannatt, F.E.S. British Museum (Nat. Hist.),
Cromwell Road, S.W. DONNINGTON, VaNBRUGH ROAD,
Blackheath, S.E.,
Wi April, 1902. To F. V. Theobald, Esq., M.A. British Museum (Nat.
Hist.),
Cromwell Road, S.W. To Walter Dannatt, Esq., F.E.S., Blackheath. Dear
Sir,—As far as I can judge, the two remains of the Culicidae
you send are those of Culex pipiens, the perfect one certainly is. No doubt
the smaller one is Gidex dorsalis, a very vicious biter. The usual times Anopheles macidipennis and different Cidex
larvae are
found is in June, July, August and September. This would be the time
to destroy the larvae, I fancy. A. hifarcatus, as I mentioned in a previous
letter,
occurs
in the
larval
stage
in winter
also. They
breed
in
rain
barrels, small puddles,
etc.,
so the work must be done very thoroughly. I should
advise
a look-out
to be kept for
the
larvte
before the
parafl&n
treatment is started. The paraffin
is best applied by dipping a bundle of rag tied on the end
of a stick into the paraffin and then dabbing the pool over in many places,
so as to get the whole
surface covered with
a
thin film. I do not think
fish would be hurt by the treatment, and doubt
if you can get a better
substitute at present for
paraffin. I will find out from Dr. Daniels, who
has bred Gidex dorsails, where they
live. The
only ones
I have found
were in a small puddle near a tap at Rochester. The
paraffin treatment, I
think, should be done two or three times at
a week's interval soon after the larvae are first noticed. I am, etc., I am, etc.,
(Signed)
Fred. V. Theobald. (Signed)
Fred. V. Theobald. 6o First Report on Economic Zoology. GEO UP
E. Animals
which
concern Man
as
causing
bodily
injury
or
disease, both
possibly
of a
deadly character,
to
(A)
his
stock
of
Domesticated
Animals,
or
(B)
his
Vegetable
Plantations, or
(C) to Wild Animals
in
the
preservation
of which he
is interested, or (D) Plants in the preserva-
tion of which he
is interested. GEO UP
E. Animals
which
concern Man
as
causing
bodily
injury
or
disease, both
possibly
of a
deadly character,
to
(A)
his
stock
of
Domesticated
Animals,
or
(B)
his
Vegetable
Plantations, or
(C) to Wild Animals
in
the
preservation
of which he
is interested, or (D) Plants in the preserva-
tion of which he
is interested. SUB-GEOUP A,
ANIMALS WHICH CONCEPtN MAN BY
CAUSING
BODILY INJUEY OE
DISEASE
TO
HIS
STOCK OF DOMESTICATED ANIMALS. * Alcohol must not be used as a stimulant after Thymol. Horse Worms and the Use of Thymol. The following enquiry was received from a correspondent at Fort
Camden,
Crosshaven,
Co. Cork, regarding
horse worms
and
their
treatment
: " Seeing
several
articles lately
in the
Field
about
the
use
of
Thymol for
the eradication
of worms
in
horses, and on the 5th a
letter from the Hon. Miss Dillon mentioning your name
as having
had a
large experience in
the
use
of Thymol
for
that purpose,
I
would
be obliged
to you
if you would give me information on the
following points:—The amount required for a mare 15*2, five years
old;
the worms
are about one
inch
long, small white
ones. The
mare has been out on grass
all the summer
till about a month ago. Should the mare be
kept on
bran mashes
for a day or two before
giving Thymol." y
The following reply was sent
: " From your
description
I imagine the worms
in your mare are
the Maw Worm {Oxyuris curvula). Thymol has been found of great
benefit
for
these nematodes,
as well
as
for
the armed
strongyles
{Sderostomum equinnm, S. tetracanthum, and
;S^. ruhnim). The
best
plan
is
to
give
bran
mashes
the
day
before and
then
give
the
Thymol
in
sweet milk. Fifteen
grains have been found
sufficient
early
in
the
morning
and
again
in
the afternoon
for
all
horses. Depluming Scabies
in Fowls. 6i Up
to
tliree drams may be given, but in
all
cases
I have known,
fifteen grain doses
are
sufficient, and more simply produce collapse. This
can
of
course
always
be
counteracted
by
administering
stimulants." * The importance
of Thymol as a nematocide
is now well-known,
it being especially valuable
for
the armed
sclerostomes that cause
often fatal epizootics in studs in this country. Depluming Scabies
{Sarcoptes
Icems) in Fowls. Depluming Scabies
{Sarcoptes
Icems) in Fowls. Amongst the enquiries
regarding animal
parasitic
diseases was
one from Mr. K. J. J. Mackensie, Lecturer on I'oultry-keeping to the
Suffolk
County
Council,
regarding
Depluming
Scabies
in Fowls. Feathers were sent from
birds
suffering from an ailment with symp-
toms pointing to this disease. An examination
of the feathers sent
did not reveal any Sarcoptes
Iccvis. This mite, which produces the
acariasis, ending in
loss
of feathers, lives
at
the base of the
quills
amongst a white powdery substance. The feathers sent had broken
off close
to
their
roots, and any powdery substance attached would
have gone. As
far as one can say without
definite proof
(i.e. the
actual presence of mites) the feathers looked
as
if the parasite had
been
at work. To
be
sure
of
this
it would be necessary
to send
feathers pulled out from the diseased bird that do not seem diseased,
and also scrapings of the skin from the invaded
areas
; such material
must be fresh or sent in
dilute
spuit. This
disease was found by
Eailliet in 1886 in a poultry-yard in Normandy, and
since in many
fresh areas. In this country there
is a general idea that " feather-
pulling " or eating is due solely to vicious habits, whereas it is nearly
always due
to the minute Sarcopt mite which lives around and
at
the base
of
the
feathers. This acarus
is most abundant in spring
and summer
;
males,
females,
and
larvae
are
all
found
together
around the leases of the feathers on any part of the body. A dozen
or more mites may often be found
in
close proximity, causing the
iiritation which leads to the birds plucking at the feathers. First Report on Economic Zoology. First Report on Economic Zoology. 62 SUB-GEOUP
B. ANIMALS WHICH CAUSE INJURY OR
DISEASE TO MAN'S VEGETABLE PLANTATIONS. Tmetocera ocellana, Schiff. a
Ocellana, Schiff. = luscana, F. ; = comitana, Hb. ; = 'pyrifoliana, C!em. ; = oculana, Harris
;
= ocuUna, Pack. Tortrix ocellana, Schiff, Syst. Verz. Schm. Eur., 130, No. 7 (1776). Pyralis ocellana, F., Mant. Ins. II., 228. Pyralis ocellana, F., Mant. Ins. II., 228. Fyralis luscana, F., Syst. Ent. (1787)
;
III. (2), 255, (1793). Fyralis luscana, F., Syst. Ent. (1787)
;
III. (2), 255, (1793). Tortrix comitana, Hb., Sml. Schm. Ent. (1800). Tortrix comitana, Hb., Sml. Schm. Ent. (1800). Grapholita (Tmetocera) ocellana, Schiff. = lariciana, Z. Stett. Ent. Ztg., XXX Grapholita (Tmetocera) ocellana, Schiff. = lariciana, Z. Stett. Ent. Ztg., XXX = lariciana, Z. Stett. Ent. Ztg., XXXIV., 129-130 (1873). Zellerana, Borgniann, Forst. N.W. Ztschr. (Tubeuf), IV., 171. Zellerana, Borgniann, Forst. N.W. Ztschr. (Tubeuf), IV., 171. Tmetocera
ocellana + lariciana,
Stgr. and
I?bl.,
Cat. Lep. Pal.,
II.,
128,
No. 2255, 9 (1901), M. Europe. Tmetocera
ocellana + lariciana,
Stgr. and
I?bl.,
Cat. Lep. Pal.,
II.,
128,
No. 2255, 9 (1901), M. Europe. {Hedya
ocellana,
Fab.) Numerous
enquires have
been made
during
the
past
summer
concerning the
larvte
of a small
Tortricid Moth which damages the
buds of various fruit trees when they are bursting and for some time
after. These
larvcC which
are
prevalent
to a greater or less extent
every year are mostly those of the Bud Moth {Hedya ocellana, Fab.). This
fruit
pest
has
long
been known
in
Europe, Kollar
having
described
its ravages
as
far back
as
1857. Since
that
date
little
has been added to our knowledge in Europe, but in 1896 Slingerland
published
a
detailed account
of
its
life-history. The accounts
of
Kollar and Slingerland differ in some essential
points, but the insect
they write about
is
undoubtedly the
same. It
is
quite probable
that both observers are right, the insect
living in two ways,
as we
see may occur in the
case
of the Colorado Beetle {vide p. 87). The
Bud Moth not only occurs in Europe, but also on the North American
Continent, both in Canada
and
the United
States. The
south of
England has been most affected by its presence, but it occurs in the
western and eastern counties. It has been especially recorded from
Bournemouth. Most fruit trees are subject to its ravages, but cheny
and apple suffer to the greatest extent. The " Bud Moth " has been
described under a
great
variety of names, some of which
are given
below
: Hedya ocellana, Fab. Penthina ocellana, Tr. Sjnlonota ocellana,
Pyralis luscana, Fab. Tortrix comitana, Hb. Tmetocera ocellana, Schiff. Penthina oculana, Harris. Penthina pyrifoliana, Clem. Hedya ocellana, Fab. Penthina ocellana, Tr. Sjnlonota ocellana,
Pyralis luscana, Fab. Tortrix comitana, Hb. Tmetocera ocellana, Schiff. Penthina oculana, Harris. Penthina pyrifoliana, Clem. The Bud Moth. 63 Tlie following references have been kindly supplied by
j\lr. Jolm
Hartley Durrant
: Tlie following references have been kindly supplied by
j\lr. Jolm
Hartley Durrant
: First Report on Economic Zoology. First Report on Economic Zoology. 0. OcELLANA, Schiff. ; + Laeiciana, Hein. = zdhrana, Brgmn. lariciana, Hein, Schm. Deutsch. Tortr., 206, No. 371 (1863). Sjnlonota lariciana, Kuaggs, Eiit. Ann., 1866, 166 (1865). Hedya lariciana,
Brit. Ent. Mo. Ma?., X., 66 (1873). Tmetoccra ocellana, Tern, Mass. Agr. Exp. Stn. Bull, 12 (1891). Timtocera ocellana,
Ltnr., Can. Ent., XXIIL,
231-2 (1891);
Kept. N.Y. VII. (1891). Tmetocera ocellana, Fletcher, Rp
Dp
Agr
Can
1891
195 (1891) 0. OcELLANA, Schiff. ; + Laeiciana, Hein. = zdhrana, Brgmn. lariciana, Hein, Schm. Deutsch. Tortr., 206, No. 371 (1863). Sjnlonota lariciana, Kuaggs, Eiit. Ann., 1866, 166 (1865). Hedya lariciana,
Brit. Ent. Mo. Ma?., X., 66 (1873). Tmetoccra ocellana, Tern, Mass. Agr. Exp. Stn. Bull, 12 (1891). Timtocera ocellana,
Ltnr., Can. Ent., XXIIL,
231-2 (1891);
Kept. N.Y. VII. (1891). Tmetocera ocellana, Fletcher, Rp. Dp. Agr. Can., 1891, 195 (1891). Tmetoccra
ocellana,
Slingerland, Cornell Univ. Agr. Exp. Stn., Ent. Div,,
Bull. 50, 3-29, figs. 1-8, III. (1893). Tmetocera
ocellana,
Slingerland, Cornell Univ. Agri. Exp. Stn., Div. Ent.,
Bull. 107, 57-66,
figs. 32-39 (1896). Tmetocera
ocellana,
Ltnr,
U.S. Dept. Agr.,
Div. Ent. (n.s.). Bull. 6,
54,
N.Y. (1896). Tmetocera
ocellana,
Sndrs., U.S. Dept. Agri., Div. Ent. (n.s.) Bull. 26,
69 Sjnlonota lariciana, Kuaggs, Eiit. Ann., 1866, 166 (1865). Hedya lariciana,
Brit. Ent. Mo. Ma?., X., 66 (1873). Tmetoccra ocellana, Tern, Mass. Agr. Exp. Stn. Bull, 12 (1891). Timtocera ocellana,
Ltnr., Can. Ent., XXIIL,
231-2 (1891);
Kept. N.Y. VII. (1891). Tmetocera ocellana, Fletcher, Rp. Dp. Agr. Can., 1891, 195 (1891). Tmetocera ocellana, Fletcher, Rp. Dp. Agr. Ca Tmetoccra
ocellana,
Slingerland, Cornell Univ. Agr. Exp. Stn., Ent. Div,,
Bull. 50, 3-29, figs. 1-8, III. (1893). Tmetocera
ocellana,
Slingerland, Cornell Univ. Agri. Exp. Stn., Div. Ent.,
Bull. 107, 57-66,
figs. 32-39 (1896). Tmetocera
ocellana,
Ltnr,
U.S. Dept. Agr.,
Div. Ent. (n.s.). Bull. 6,
54,
N.Y. (1896). Tmetocera
ocellana,
Sndrs., U.S. Dept. Agri., Div. Ent. (n.s.) Bull. 26,
69
(1900). LITERATUEE. Pentliina pyrifoliana, Clem., Pr. Ac. Nat. Sc. Phil., 1860, 357 (1860). Penthina oculana, Harris, Inj. Ins., 482 (1862). Pentliina oculana. Pack., Rp. Mass. Bd. Agr. (1869); Am. Ent., I., 251. Grapholitha oculana, Sndrs., Can. Ent., III., 13, fig. 9 (1871). Grapholitha oculina. Pack, Mass. Agr. Rpt. (1870). Grapholitha oculina. Pack, Am. Nat., IV., 684 (1871). Tmetocera ocellana, Z., Beitr., 61 (1875). Tmetocera ocellana. Fern., Tr. Am. Ent. Soc, X., 48, No. 349 (1882). Tmetocera ocellana. Fern., Tr. Am. Ent. So Tmetocera ocellana, Fletch., Rept. Dep. Agri. Can. (1885). Tmetocera ocellana, Harvey, Ann. Rp. Marine Exp. Stn. (1888). Tmetocera ocellana, Tern, Hatch. Exp. Stn. Rp., 1888, 11-12 (1889). Tmetocera ocellana. Cook, Ann. Rep. Mich, Agri. Exp. Stn., IV. (1891). Tortrix comitana, Hw., Lp. Br., 434 (1811). Tortrix comitana, Hw., Lp. Br., 434 (1811). Tortrix luscana, TroL, Enum. Tortr. Wiirt., No. 106 (1828). Penthina luscana,
Dp., H.N. Lp. Fr.,
IX.,
203,
pi. 245,
10 (1834), and
Supp. IV., pi. 84, 2, Penthina ocellana, Tr., Schm. Eur., X. (3), 51. Penthina comitana, Wd., Ind. Ent. (1833-9). Penthina ocellana, Tr., Treat. Ins. KoUar, 234 (1837). Tortrix (Penthina) ocellana, HS. and SB., Schm. Eur., IV., 233 (1849). Hedya ocellana, Wilk., Br, Tortr., 118 (1859). Tmetocera ocellana, Ld., Wien Ent., III., 367-8 (1859). Grapholitha (Tmetocera) ocellana, Hein., Schm, Deutsch, Tortr., 206 (1863). rapholitha (Tmetocera) ocellana, Hein., Schm Grapholitha (Tmetocera) ocellana, Hein., Schm, Deutsch, Tortr., 206 (1863). Tmetocera ocellana, Stgr., Cat. Lp. Eur,, 260, No. 1210 (1871). Tmetocera ocellana, Stgr., Cat. Lp. Eur,, 260, No. 1210 (1871). Grapholitha (PxcUsca) ocellana, Snell, Vlind. Ned. Micr., 325-6 (1882). Tmetocera
ocellana, MP. and FT.,
Nat. Sc,
VHI.,
188
(1889);
Meyr. H. B., Br. Lp., 476 (1895). Tmetocera ocellana, Stgr. and Rbl., Cat. Lp. Pal., 11., 125, No, 2255 (1901)., Tmetocera ocellana, Stgr. and Rbl., Cat. Lp. Pal., 11., 125, No, 2255 (1901).,
Hedya ocellana, Theob , Agri. Gaz., 7
7
and 14. 7 (1902) Hedya ocellana, Theob., Agri. Gaz., 7. 7, and 14. 7 (1902). 64 How THE Test may be Detected. The
presence
of this
fr\iit
pest may be
detected
by
the
early
destruction of the developing buds, which on partly opening are seen
to be eaten and
shrivelled and which soon
t\vn\ brown
; these buds
will be found
to contain a small reddish-brown
caterpillar, the cause
of the injury. Later the damage is still more noticeable, the opening
leaf and blossom being spun together, for both leaf and blossom buds
are attacked. There may frequently be noticed a gummy appearance
of the opening
leaflets. The
larva
enters
the
bud,
if
it
has
not
already
burst, by
eating down
between
the
bracts, and
there,
as
described by KoUar, a drop of sap forms which tends to hold the bud
intact and
to stop it from bursting
;
in any case if
it does open the
young leaves soon shrivel and turn brown. During the past summer,
trees, especially cherry, have been quite ruined in
this way. More
usually, many of the buds fully open, leaf and blossom appearing as
usual,
the
caterpillars
later
spinning
the
bunches
of
leaves
and
blossoms into a mass with silk. These leaf nests frequently die right
away and turn brown in a very characteristic manner. Slingerland
states
that in America the larvte
also sometimes " burrow down
the
stalk for two or three inches, causing
it
to die."
This habit has not
so far been observed in England
;
in all cases the larva boring down
the top shoots of apple and pear has proved
to be that of the
l*ith
Moth {Laverna atra) (vide
p. 68). When nearly mature the Bud
Moth
larva forms more
or
less
of a tube by rolling up a leaf and
roughly
lines
it with
loose
silk. The
caterpillar at
first only uses
this
tube
as
a
shelter
from which
it
emerges
to
feed,
gradually
drawing
the neighbouring
lea^'es
together
by
silken
cords. From The Bud Moth. 65 ol)servations made during the last few years
it seems that the cater-
pillars chiefly feed at night. Life-history. The Bud Moth (Fig. 5, d) is one of the Tortricidce. It
varies
in
length from half-an-inch
to two-thirds
across
the
expanded wings. The front wings are dark grey with a broad greyish-^^'llite band across
the middle with grey spots and streaks
; near the anal angle of the
wing is a triangular
blackish spot and towards
the
tip a leaden-grey
eye-like spot with several black dots. The hind wings are grey. The Fig. 5,
the bud moth (H. ocellana). a, The lava
;
b, the pupa
;
c, leaf eaten by larva; in autumn
;
d, imago
;
e, eggr (after Slingerland). Fig. 5,
the bud moth (H. ocellana). a, The lava
;
b, the pupa
;
c, leaf eaten by larva; in autumn
;
d, imago
;
e, eggr (after Slingerland). fore wings
are subject
to considerable
variation
;
in some specimens
the median greyish-white band is distinctly dull slaty-grey. This moth appears in June and July when it may be seen flying
about in orchards, gardens, and lanes at
dusk. During the day they
rest amongst
the
leaves
of the
trees and
upon
the
trunks,
their
coloration
rendering
them
most
inconspicuous,
especially
when
resting on an old moss or lichen grown tree. The female lays her eggs at night
; they may be deposited singly
or
in
clusters, usually upon
the upper sides
of the
leaves. Kollar
states, however, that they are placed at the base of leaf and fruit buds
and
that they remain as ova during the whole of the winter. Both r 66 First Report on Economic Zoology. Fletcher and
Slingeiiand have found
that
the
eggs hatch
in
late
summer and
early autumn and
that winter
is passed in the
larval
stage. This is undoubtedly the usual
case, but
that some hibernate
in
the egg condition
I think very probable, as I have found young
larvffi in the early spring not more than one-twelfth of an inch long,
whilst those that hibernate are considerably larger. The
eggs
(Fig. 5,
e)
are
flat, round
or
oval,
the
centre
being
slightly elevated, they are very transparent and look like little drops
of gum on
the
leaf. When
several are
laid
together they overlap
one another like flsh
scales. Food Plants. All
fruit
trees
are
attacked
by
this
pest, but
it
is
especially
cherry and apple that suffer in
this country. It also occurs on the
sloe and plum, and has been recorded from the blackberry, whilst in
North America it attacks the peach and quince also. Life-history. In
general appearance they resemble
the ova of the Codling Moth
;
the outer edge is marked with a well
defined reticulate sculpturing
; the
central part of the egg is usually
green, the colour being due
to the developing larva within showing
through. The egg stage lasts from seven
to ten days. The young larvse on
coming from the egg at once commence to feed upon the lower layers
of the leaf, forming
for themselves a
little tube of silk open at each
end and attached to the
leaf,
usually at the mid-rib. When feeding
off the
leaf the
larvse form a
slight silken web
which serves
as a protection for them. In this
manner the larvse go on feeding until about the
middle
of September, by which time they
are
about half grown. Professors
Slingerland and
Fletcher were the
first to show that these small
larvse pass the winter in small silken cases on the
trees, as inconspicuous objects covered with dirt
and not more than one-eighth of an inch long. These winter
houses
(Fig. 6) are
found
at
the
base of a bud or under a dead leaf or bud
scale. When quite young
the
larvse
are
green, and
this
is
the
colour
most
usually
found when
they
are
in
their
"houses."
When
the buds
commence to
swell
they
crawl
out
and
enter
them and become dull reddish-brown with black
head
and
black
first
segment
;
when
nearly
mature they become a more pronounced reddish-
brown,
the
dark
head,
first
segment
and
legs
showing
up
pro-
minently. When quite
mature they reach half-an-inch
in
length. During the
latter part. of
their
life they
feed
amongst
the
leaves,
which
they
spin
together, and
pupate
in
a
tube
of
dead
leaves
as previously described. The pupa
is
bright brown, with two rows
of backwardly projecting spines on each segment. Fig. 6. Twig with larval cases (a) of
Bud Moth and buds (6). Fig. 6. Twig with larval cases (a) of
Bud Moth and buds (6). Fig. 6. Twig with larval cases (a) of
Bud Moth and buds (6). g
Twig with larval cases (a) of
Bud Moth and buds (6). The Bud Moth. 67 One
brood
only occurs
in
Great
Britain. The moth
is
very
common
all over the South
of England, but becomes
rarer
in
the
North. Natueal Enemies. Five species of Ichneumon flies prey upon the caterpillars of this
moth in Europe, but none have been noticed
in Great
Britain. In
North
America
they
also
are
preyed
upon
by
three
species
of
Ichneumons. Amongst birds we find the blue and great tits {Parits
cceriUeus and F. major) picking the
larvae out of the buds and leaf
nests. The sparrow also has been observed feeding upon them. A
large sand-wasp,
Odynerus
catsJcillcnsis,
stores
its
nests with
these
caterpillars
in
North
America. None
of
these
natural
enemies,
unless
it be the Paridte or Tits, do much good in keeping down this
Bud Moth. (Antithesia
varier/ana, Hb.) Another Tortrix
lives
in
a very similar way
to
the Bud Moth,
namely,
Antithesia
variegana
or
A. cynosbatcUa. This
moth
is
about
two-thiixls
of an inch in width of expanded wings
;
the
fore
wings have the
basal two-thirds brownish-black, the apical
third
is
white
clouded with
grey towards
the hind margin
;
there
are also
two or three black spots projecting from the edge of the central band. The moth appears in June, and lays her eggs on the leaf. The larvae
are
tliick
and
dark
green
with
black
spots
;
the
head
and
first
segment
also being black. They may be found in the early spring
on hawthorn and
sloe growing in the
hedgero'S\'s and
also in most
apple and
pear orchards. The winter
is
passed, according
to
Mr,
Newstead, much in the same way as the Bud Moth. Penthina pruniana, a closely related
species, also probably lives
in the same way. Penthina pruniana, a closely related
species, also probably lives
in the same way. Methods of Preventing the PiAvages of Bud Moth Laev.e. Now that we know
that the larvae feed upon
the
leafage
in
the
late summer we can
to a large extent check the ravages of this pest
by
arsenical
spraying. Larvae
are
always
more
easily
destroyed
when young, and there
is not the
least doubt
that spraying
in
the
autumn will
kill them. There should
also be a second washing in
the
spring when
the
caterpillars
are
to some
extent
exposed
just
when the buds are bursting, and this followed by a third dressing
to
kill those that escape when they are in their leaf and blossom nests. Hand-picking may be resorted to in gardens and nm-series and where
single low
trees
are invaded, the
leaf
nests
being
easily seen and
picked off by hand
before
the moths have emerged from
the pupal
stage. Washing with
caustic
alkali wash does not seem
to check
this
pest,
for
trees
so
treated
last winter (1902)
suffered severely from
the Bud Moth and also Pith Moth. Probably the
larval
cases were
hidden under the bud-bracts and in such places that the wash does
not reach, the larval cases also protecting the caterpillars within from
the burning action of the wash. F 2 First Report on Economic Zoology. 68 The Allied Bud Moth. The Allied Bud Moth. (Antithesia
varier/ana, Hb.) Life-history. This moth belongs to the group of small moths known as Thidnm
and
to the genus Laverna. Its wing expanse
is a
little
less than
half an
inch when
fully expanded
;
the
front
wings
are
almost
entirely
black, but may
be
mottled
with
black, dark
brown and
rusty brown
; the inner margin of the fore wings
is- white
to beyond
the middle, where an irregular oblique white bar proceeds
to the
tip
of the wing, and two branches from
this
intersect the black
apical
portion
; the
posterior wings
are
grey
and,
like
all
Tineince, have
long
fringes;
the head
is
almost
entirely
white. It
is
subject
to
much variation. Some specimens
are almost black
;
these Stainton
considers a
distinct
variety. The moth appears in June according
to
Stainton, but all those that have been bred or observed appeared
in
July. Some received
in 1898
hatched on July
5th, others not
until
the
21st. Difference
of
locality
is
sure
to
account
for
a
difference in the time of their appearance. The eggs are apparently
laid on the leaves
; no definite observa-
tions
have
been made, however. In July
I
found
several
small
batches of eggs on an apple
tree previously badly attacked by Pith
Moth, but I am not certain if they were those of the Laverna. They
occurred
in small
batches from
one-fifth
to
one-fourth of an
inch
long
;
in form they resembled those of the Bud Moth, ^dz., flat and
scale-like and
almost
transparent. One
batch
was
composed
of
twenty eggs
overlapping
one
another like
fish
scales. The whole
surface of the egg
is covered with a well-defined reticulate sculptur-
ing, not the outer part only, as in those of the Bud Moth or Codling
Moth. Penthina variegana was seen near
this
tree, so that
it may
be the eggs were of that insect. In any case the eggs, whatever they may be like, give rise
to
the
larvae the same summer, and the young larvae feed first on the leaves. As winter approaches,
the
larvte, wliich
are
still quite
small, bore
just under the bark of a twig or into the apex of a shoot and remain
there most of the winter. During January and February the
larvse
tunnel right
into
a young shoot and work up
the
pith (Fig. 8,
e). (Lavema
atra, Haw.
; 23utri]pcnella, Zell.) (Lavema
atra, Haw. ; 23utri]pcnella, Zell.) Numerous inquiries were made during the past year concerning
the
larva
of a small moth—the
Pith Moth {Lavema
atra). This
attack has been known
to
fruit-growers and
gardeners
for
a
long
time. It
is
noticed every
year,
but
in
1902
it
was abnormally
abundant, and
a vast amount
of damage was done by it over Great
Britain,
The moth
is not
often
seen, and probably but few people
are acquainted
with
it. It
occurs
over a
large
area
of
England
up to Newcastle. The
damage
done
by
the
larva? is
particularly noticeable
in
nursery stock. They burrow up the terminal
shoots and
kill them,
the
result being deformed and stumpy
trees. The attacked shoots
flag and then die and turn brown, the dead masses varying from two
to four inches
in
length. These dead shoots may remain some time
on the tree, or they may fall to the ground naturally, or be beaten off
by heavy rain. The attack may readily be told from that of the Bud
Moth by the
absence
of leaves spun
together, and the absence of
damaged buds and blossom
; but the whole shoot dies away. Apple
trees
are
chiefly attacked, but reports
of
its
ravaging
pears
have
also been received. The larvte are also found on hawthorn and other
wild Eosace.ie. Although it is a wide-spread
pest, the following
localities may be The Pith Moth. 69 mentioned as having been particularly troubled with
it
: Worcester
Herefordshire
(Eoss)
;
Gloucester
;
Cambridgeshire
(Wisbech)
;
Bournemouth
;
Sussex
(especially
at
Polegate)
;
Surrey and
Kent
generally. Life-history. In
this
tunnel the
caterpillar
lives
until June
;
its presence
does
not stop the leaves and blossom from unfolding, although later they
flag, turn brown, and
die
right
off" (Fig. 7,
b). These dead shoots First Report on Economic Zoology. 70 Fig. 7. the pith moth (Lavcrna atra). A, Imago
; B, attacked apple-shoot, the upper portion shrivelling up and dying away
'
C, processes on pupa
; D, pupa (enlarged) in situ
; E, showing position of larva
(natural size) in situ; F enlarged larva in broken open bud
; G, larva (enlarged). Fig. 7. the pith moth (Lavcrna atra). A, Imago
; B, attacked apple-shoot, the upper portion shrivelling up and dying away
'
C, processes on pupa
; D, pupa (enlarged) in situ
; E, showing position of larva
(natural size) in situ; F enlarged larva in broken open bud
; G, larva (enlarged). Fig. 7. the pith moth (Lavcrna atra). A, Imago
; B, attacked apple-shoot, the upper portion shrivelling up and dying away
'
C, processes on pupa
; D, pupa (enlarged) in situ
; E, showing position of larva
(natural size) in situ; F enlarged larva in broken open bud
; G, larva (enlarged). A, Imago
; B, attacked apple-shoot, the upper portion shrivelling up and dying away
'
C, processes on pupa
; D, pupa (enlarged) in situ
; E, showing position of larva
(natural size) in situ; F enlarged larva in broken open bud
; G, larva (enlarged). Th
Pith M th The Pith Moth. 71 it" broken
off will be found
to contain the Pith Moth caterpillar or
pupa, usually situated near the apex of a shoot. The
larva (g) is dull reddish-brown with a deep brown head and
first segment
; the other segments show more
or
less
traces of pale
brown spots four
in
a row on
the second and
third segments and
four placed
in
a quadrangle on
the remaining segments. The two
anterior segments have two
lateral spots and the remainder a single
lateral spot. The apex is deep brown. When mature they reach one-
third of an inch in length and then pupate near the apex of the shoot
they have tunnelled. They reach their full-fed stage during the last
two weeks in June. If the dead
shoots are picked off, the moth can
be easily bred. Life-history. The pupa (d) is of an ochraceous hue
;
the head and front
of the
thorax and tip of the body mahogany red. It is cylindrical in form
and about one-fourth of an inch long. On the ventral surface of the
penultimate segment
are two blunt processes separate and diverging
outwards, hairy at their apices (Fig. 7, c)
; the eyes are black and the
wing cases and
legs
long, the former pointed. This stage lasts from
two
to
three weeks, the moths emerging from the end
of June into
July. The pupas may sometimes be seen
projecting from
the dead
shoot. Stainton says the
larvre
also occur
in hawthorn
berries
in
September and that the black variety only is found
in apple shoots
in Februaiy and March. Preventive Measuees. It is quite obvious
that there can be no remedy for this pest, but
we can do much in the way of prevention by hand-picking the dead
shoots
before the moths emerge
in June. This can only be done, of
course, where small trees are attacked, and it usually happens that it
is only on such trees that the attack takes place. Late spraying with Paris green would probably prove
beneficial,
as it would
kill the young
larvse, which seem to feed
first of all on
the
leaves. Of course, care must be taken in regard
to
the fruit. The trees should be washed not less than four weeks before the fruit is
gathered, but as soon as it is harvested a heavy spraying may be given. The time to spray therefore must depend on the variety of apple
concerned. This
is
certainly worth
giving
a
fair
trial,
as beyond
hand-picking we can do nothing to check the increase of this pest, so
noticeable during the past few years. First Report on Economic Zoology. 72 The
Pear and
Cherry Sawfly
or Slugworm.
{Eriocamjpa limacina, Cameron.) A few enquiries were received during the past year regarding the
Pear and Cherry Sawfly {Eriocanipa
limacina). This fruit
tree pest
has not been
so abundant
as usual
during
the
past summer and
autumn. In
some
districts
where
it
is
usually harmful
it
has
scarcely been
noticed. One
correspondent
^WTiting
from
Sitting-
bourne, Kent, asked
for information
concerning these
pests, "found
in numbers on and destroying his plum and cherry trees," and for the
best means of preventing and destroying them. Then- normal food
plants
are
cherry, pear and sometimes hawthorn. It has not been
notified before as injurious to plum, although Miss Ormerod mentions
plum, and
sometimes
peach, as being occasional food plants, and on
one
occasion it is recorded on the quince. Cameron, in his work on
" British
Phytophagous
Hymenoptera "
(vol. i. p. 225),
mentions
other food plants, as Eubus, Amygdalus, Quercus and Betula. There
is no doubt
that
this insect
is very
susceptible to damp
weather and thus has not been nearly so harmful during the past
year. Considerable relief from
this
pest has been reported by adopting
the plan of removing and burning the surface
soil during the winter
months from beneath trees that had been attacked. Phyllobius and Insects on
Vines. 73 To this the following reply was sent
: From the
fact that
yon
sprayed
twice
with
Paris
green, and
the
description you give of the
withering
shoots, leaves, and
flowers in your
orchards, I feel pretty certain you have been suffering from the combmed
attack of two
of the prominent apple and pear pests this season, viz., the
Pith Moth {Laverna
atra) and the Bud Moth
{Hedya
oceJlana). The
former
is most abundant
generally, but
I have had many reports of the
damage done by the latter. g
y
The Pith Moth is in the pupal
stage in the dead shoots
still and wiU
soon hatch out. On
a small
scale these should be hand-picked, but
I
doubt
if
it could be
done
in
large
orchards. But
judging
from
the
recorded
facts
in
the
life-history
of
these two
moths,
I am
strongly
inclined to
believe we can cope with them on
a
large
scale by autumn
and
late summer
spraying,
for
the
eggs
hatch
out
in July and the
larvie
(small)
feed
until
the
winter, when
they
hibernate. Arsenical
spraying then, say in August (depending on the
fruit), and again
as soon
as the crop is harvested would give relief, for at no other time can we get
at the Pith Moth, and not at the Bud Moth until
it has done the damage. The green caterpillars you refer to,
if they are not " loopers," are probably
Penthina variegana, one of the Tortrices related to the Bud Moth.* Phyllobius or Leaf Weevils. A single instance only of the
attack
of Leaf Weevils
has been
reported. In
districts where they are usually very abundant they
have
only
occurred
in
small
numbers
during
the
past
summer. Messrs. James Carter & Co. sent specimens
of the species Phyllohiiis
viridaris on the 19th of June, stating that they were overrunning the
garden of a
client, and asked
for information how
to eradicate the
pest. This particular leaf weevil is common on elm hedges as a rule. The following reply was sent
: With regard to eradicating this pest, the only treatment is as follows
:
1. Spraying with Paris green wherever the foliage
can be so treated
BOW (i.e. on apple, plum, pear, and nut). 2. Beating down the
beetles on to tarred sacks
in early morning has
been found to clear a garden speedily of this pest. But probably spraying will be
the
least
costly treatment if on a large
scale. *
Since
this
report was
sent, I have found the treatment advised has
cleared
Apple trees of the young larva?. Notes on Fruit Pests
in Orchards at Wisbech. Some interesting notes on
the ravages of insect pests were sent,
together
with an
enquiry
as
to
the
cause
of the
damage,
from
Mr. B. W, Gatherwood, of Wisbech. In this letter he states that Plum blossom was cut
off by frost, but apple
trees
mixed
with
the
above were comparatively all right, except for a few caterpillars, of what I
took to be the Winter Moth, on the 24th of May, the trees showing every
sign of
a plentiful crop
of apples,
I may say in the last week
of April
and the first week in May we syringed twice with Paris green (1 in 200). When I returned home on the 7th
of June my apple
trees and some of
the plum presented an appearance as if a hot blast of
air had passed over
the whole garden, withering all
shoots, leaves, and flowers
; the few leaves
left were all riddled with holes, leaving only the ribs of the leaf,
I could
find no insects then
or since except a few green caterpillars,
I am quite
at a
loss
to know the cause
of
this wholesale
destruction. I should be
glad
to have a reply from you on the
subject, and you would
be
con-
ferring on the
district a great boon
if you
would
suggest
a
remedy,
I
firmly believe
if we
had kept dressing
the
trees
with
the
solution
mentioned until the apple blossom had gone we should have had a crop. Phyllobius and Insects on
Vines. Insects on Vines in Jersey. Three kinds
of
Arthiopods have been sent
by
Col. Sanderson,
from
Jersey,
attacking
the
vines, with
enquiries
regarding them. They were the following
: First Report on Economic Zoology, 74 (i.) Two small larval mealy Bugs {Dactylohius citri). (i.) Two small larval mealy Bugs {Dactylohius citri). (ii.) A single mite {Tetranyclius telarius). (ii.) A single mite {Tetranyclius telarius). (iii.) Several specimens in larval, pupal, and adult winged stages
of one of the Pseudo-Neuroptera belonging to the family Psocidce. The following note was sent to Col. Sanderson
: The life-history of the Psocidce is roughly as follows
: The females
lay their eggs in patches on leaves, bark, etc., of trees and plants and
other
objects, many on
dead and
decaying
matter
and
preserved
objects. The females cover the eggs with a web. Larvoe and pupfe
are much alike
; wing-buds gradually appear on the larva and so the
pupal stage is assumed. Dry vegetable matters and lichens form the
food of one section (PJiocina)
; the food of the other section {Atropina)
is dried insects, plants, books, papers, etc. One of the latter, Atropos
divinatoria, is sometimes
called the Death Watch,
A beetle, Ano-
hium
tessellatum, is
also
called the Death Watch. The
Psocid?e do
much harm
to papers,
books, and insect
collections. The Phocina
live more
or
less
in
societies on
tree trunks, palings, and amongst
rough herbage and on
trees, especially on Conifers
; both sexes can
spin a web of
silk. Some species do harm
to living plants, but the
majority do not. The Psocidte sent belong to the genus
Ccecilius. With
regard
to
the
male
" Mealy
Bugs "
which
you
enquire
after, they can soon be told, for they have one pair of wings as in all
other Coccidte, and thus differ from the winged Psocids. The common " Mealy Bug " is known
as
Dactylohius
citri, Bois-
duval, and is the same as D. dcsiructoi^ of Comstock. It is a world-
wide
species. The males
are not
so very
rare
; they are
of a dull
reddish
colour and have two long white thread-like processes at the
end of the body
;
the single pair of wings are dark iridescent blue,
and when the
insects
are
settled
these wings overlap and hide the
abdomen. They are very different in appearance from the sedentary
females. Insects on Vines in Jersey. Two
other
" Mealy Bugs " occur
in
Great
Britain, namely the
Long-fringed Mealy Bug (D. longipinus) and
the native Laburnum
and Gorse Mealy Bug
(Psciidococcus
ulicis). The former can be
told
by the long margined processes. The latter is really non-injurious. The
other forms you
sent
with
the
winged
Ccecilius
were
its
larval and pupal stages. Many of the Psocida3 remain very like the
larval stage you send,
i.e. in an apterous condition. The Mussel Scale. IS The Mussel Scale or Oyster-Shell Back Louse.
{Mytilasjtis pomorum, Bouche.) Numerous enquiries
are constantly being made by fruit-growers
and gardeners
concerning
the Mussel
Scale
(Mi/tilasjns jJomorum). There being no
leaflets issued
or other ready information
for
fruit-
growers,
the
following
notes
have been
prepared with
a
view
to
meeting this want. The Mussel
Scale
is found
chiefly on
apple, but
also on
pear,
currant, plum and wild
Crataegus, such
as the Hawthorn. I have
also
found
it
abundantly
on
Blackthorn
in
Devonshii-e. This Fig 8. mussel scale {Mytilaspis pomonim). a, Upper
;
b, lower siu'face of ? scale
;
c, <f scale. (Greatly enlarged.) Fig 8. mussel scale {Mytilaspis pomonim). a, Upper
;
b, lower siu'face of ? scale
;
c, <f scale. (Greatly enlarged.) pest, the worst
scale insect we have
in
Great
Britain,
is
found
in
North
America, and
also
in
Australia, New
Zealand, and
South
Africa, having been imported on nm:sery stock. In this
latter way
it
is
also
largely
distributed
in
this
country. The
necessity
of
fumigating young stock
before planting
is
thus
rendered essential
and should be done by
all nurserymen before their stock
is sent out. A few
scales may
easily
escape
detection
and
so
set
up
a
large
colony, to the detriment and even death
of the tree. I know of no
district in England where
this pest does not occur in greater or
less
abundance. Old
trees and neglected orchards
chiefly encourage
it,
but young stock suffer far more than old. This scale insect is frequently taken for growths on the bark. This scale insect is frequently taken for growths on the bark. The
scale
is
the
product
of a minute
insect belonging
to
the The
scale
is
the
product
of a minute
insect belonging
to
the First Report on Economic Zoology. 76 Coccidcc ; the male and female
scales
differ in appearance and
size
;
the male scales are seldom observed. The female scale (Fig 8, a and
h) is about one-eighth of an inch
long
;
in form the scales
are rounded behind, but taper to a point at
one end—the head end
; they may be straight or curved, and even much
contorted. In colour they vary from deep brown to almost grey. The
male scale is much smaller than
the female and of the form shown
in fig. 9,
c. The Mussel Scale or Oyster-Shell Back Louse.
{Mytilasjtis pomorum, Bouche.) They damage the trees by sucking out the sap by means
of long, flexible mouths which they insert into the plant tissues. This
scale not only occurs on the trunk and boughs of the trees, but also on
the leaf and
fruit. Foreign apples
are frequently imported covered
with
this and
other scale
pests. The
scale, as in
all
Coccidce, is
a
product
formed by
the
insect
which
lives
beneath
it,
partly
by
excretions from its body, partly by the
cast skins
of the
insect, the
so-called cxuvia:. Treatment. The
trunks, etc., of
all
trees must be
kept
clean,
i.e., free from
rough
bark, moss and
lichens. This
can be
done by washing
in
winter with
caustic
alkali wash, which
at the same time
corrodes
and loosens the
scales from the
trees. Trees badly infested should
also be sprayed in the early summer about the middle of June with
paraffin emulsion, two
or three
times,
at intervals
of
a
few
days. This kills numbers of the young and corrodes away to some extent
any remaining scales. "VA^iitewashing the trunks of the trees as far
as the forks of the boughs does some good and keeps the wood in a
healthy state. All young stock should
be
treated
to
destroy
the
scale before being planted
or soon
after. The best method is fumi-
gation with hydrocyanic acid gas, the most valuable scale remedy. Natural Enemies. Scales have many natural
enemies, but
this
species and those
that attack the currant
in
this country are not materially lessened
by them. Anyone
trusting
to
parasites
to help
the farmer in
this
respect evidently is not acquainted with these
pests in our orchards. Amongst the natural enemies birds alone do any good. The Paridae
or
Tits
feed upon
this
scale, and a few
other
birds on
the
Tree-
creeper and Wryneck. Lady-birds and
their larvae
eat
scale,
but
none
seem
very
partial
to
the
Mussel
scale
in
Great
Britain. Minute
hymenoptera
Chalcididae—also
live
as
parasites
upon
them, but
seldom do any
appreciable
good;
in
the
first
instance
they occur too late in the attack
to prevent the damage, and never
are
sufficiently abundant
to check the pest for the following year. Sound advice
to
fruit growers
is
to go on washing and ignore the
infinitesimal help given by these minute
parasites. Also encourage
those useful birds the Tits in orchard and garden. Life-history. The eggs (Fig. 3, a'), are laid by the sedentary female under the
scale. They resemble to the naked eye small whitish dust. As many
as eighty may be counted under a single scale, but the number varies
considerably. The eggs give rise
in the early summer to very small
active
six-legged
larvae, which crawl from beneath the
scales and
may be distributed from tree
to
tree by
the
wind, by
birds, and by
predatory insects, such as lady-l)irds. They are about one-hundredth
of an inch long. In a short time they fix themselves to the plant by
their short proboscis and draw away the sap
; the scale then commences
to form by the excretion of a few waxy threads and gradually grows
to the form shown in Fig. 8. During this period the larva loses its
legs and becomes converted into a fleshy legless creature
; the female
remains feecUng beneath the scale and is provided with a long flexible
proboscis, which
is inserted into the
tissues of the
plant. Towards
the end of the summer she deposits her eggs and
dies, her shrivelled
skin remaining beneath the scale. If the larva
is going
to become a male, not only
is a
different
scale produced (most
often upon
the
leaves), luit a
totally different
mature
insect. The
male
undergoes
a
kind
of
pupal
stage and
escapes from the
scale as a small winged
insect, provided with two
rather large wings and a pointed process
at the end of the abdomen,
which
it
can
insert
under
the
female
scale
and
so
carry
out
fertilisation. The males are very rare, most of the females reproducing
asexually. A single brood normally exists in this country. Mussel Scale and Beetle Mites. 77 Treatment. Soft soap wash
as suggested by a correspondent would be
quite
useless. The Oiibatida? have very hard chitinous skins, and probably
no wash that could be used in the summer would
affect them. Late
in the autumn
caustic
alkali wash might be tried and might prove
beneficial. The method of preparing and using
tlie alkali wash will be found
in Leaflet 70 of the Board of Agriculture. Oribatidae or Beetle Mites on Forest and Fruit
Trees. The
Beetle
Mites
are
frequently
sent
by
fruit-growers
with
enquuies as to their economy. The records
of them on
forest trees
are few. Specimens
haA'e
been
sent
from
the
Dii'ector, Royal
Botanic
Gardens, and recei^'ed by him from Mr. A. S. Birknell, of Barcombe,
Sussex, which are recorded by him as damaging chestnut (horse) and First Report on Economic Zoology. 78 lime trees. There are a number of species of Oribata
; one, 0. glohata,
is
often
present
in
swarms
on
fruit
trees,
especially the
plum. Instead of finding them injurious, they appear to be either beneficial
or non-obnoxious. They have been seen feeding off the spores of the
canker fungus and various green vegetal matters on fruit trees. But
in one
or two instances
fruit-gi'owers have noticed
that they have
done some harm
to
the
leaves. They often swarm in the forks
of
the boughs and
axils of the twigs and
buds. The species sent by
Mr. Bii'knell
has
been
identified
by
Albert
Michael
as
Oribata
orhicidaris, which also occurs on various fruit trees in Kent. This
species has been
dealt with
in the Journal
of
the South-
Eastern Agricultural College (No. 6, p. 11 (1897)). Albert Michael, the
chief authority on Mites, agrees that these Oribatidse do no practical
harm. Several fruit-growers
have informed me that they cause the
leaves of the plum to die off, however. It is possible this may have
been due
to
other
causes. All
cases personally investigated
have
shown no damage to the trees, although thousands of these shiny mites
were present. {Eriophyes jpyri, Sch.) Several enquiries were made concerning
the
Pear-Leaf
Blister
Mite
during
1902. Information was
sent
that
these
little
acari
enter the leaf by the stomata and
live
in
the
soft internal tissues,
where they soon commence to form a galled patch. They move from
leaf
to leaf, but spread very slowly. Often one
tree in an orchard
will be attacked for years before any neighbouring ones show signs of
having contracted the disease. Frequently we see a single branch of
a tree diseased year after year and yet the
lest of the
tree remains
clean. In nearly all cases I have
seen, the fruit becomes hard and
gritty and is usually deformed. All we can do
is
to hand-pick the Big Btid Mite and Hover Flies. 79 diseased leaves
in
July, not later, and wash the trees
frequently in
early spring and autumn with liver of sulphur wash—that is paraffin
emulsion and
liver of sulphur. Dr. Nalepa informs me the
mites
winter in the buds. The
*'Big Bud"
Mite.
{Eriophyes ribis, Nalepa.) A correspondent from near Tewkesbury
wrote
as
follows
:
" A
dealer has
offered me
6,000
black
currants
(Baldwin's), but
they
contained a lot of swollen buds like the
enclosed. I want to know
if they are infested with
Gall Mites, if so whether
it would be safe
to plant them."
The following reply was sent
:
" The black currant
buds
you
send
are
badly
infested
with
the
Currant Bud
Mite
(^Eriophyes {Phytiyptus) ribis). This mite
is perhaps more prevalent
in the Baldwin currant than in any other variety, but the only kind
I have never seen attacked
is the old cottage-garden
one, that
is
so
prevalent in Kent, a light cropper, however." " It
is most unwise to have any infested
plants, however cheap. It is really quite useless attempting black currant growing with
the
' big bud
' pest in the
plantations, as at
present there
is no known
remedy for it. It is most essential to be
successful
to
start on land
new to black currants and with clean stock." The following reply was sent
: The following reply was sent
: The
so-called
" Monkey-peas "
or woodlice
are
omnivorous
feeders. They sometimes do considerable harm out of doors
as well
as in green-
houses, especially to
soft
fruits. I have never heard
of them damaging
hops in any way, but
it is quite probable that they would do so if present
in suificient nmubers. They are more
likely to eat away on the surface of
the ground than upon the
bine. Strawberries
are
often
gnawed
away
around
the crown by them. Woodlice can
easily be trapped by putting
old baskets full
of damp moss upside down, beneath which the woodlice
congregate and can easily be collected. Woodlice
in Hops. Specimens
of the so-called Monkey-peas were forwarded on the
2nd of June from Macknade, FaA'-ersham, by Mr. F. Neame, with the
following note
: I noticed the other day in one of the hop gardens large numbers of the
insect commonly known
as
" Monkey-pea."
Are they
likely
to damage
the bine
at
all by biting
it ? If you could inform me on
this point, I
should be much obliged. Hover Flies {SyrjMdce: Aphis Feeders). A number
of enquiries have been made
this season (1902) con-
cerning
the
leech-like
larvre
of the Hover
Flies
(Syrphidce). The
following note was
sent
to Mr. Bear, of Hailsham, in answer
to an
enquiry as to the nature of these larvse
: The three larvje you send feeding on the Apple Aphis are those of one
of the Hover Flies
{Si/rphidrr.) Catabomha pijrastri, Linn. They are most
beneficial, being ravenous Aphis feeders attacking aU kinds of " Dolphins."
There are a good many species, sufficiently abundant to do a great dea]
in keeping Aphis
in check, but unfortunately they come rather
late and
much harm is often done by the Aphides before they make
their presence
felt. Syrphus
ribesil,
Linn., and
S. grossulariae,
Meig., seem
to be
the
most abundant after the one you send. g
p
g aU
ds of
Dolphins. There are a good many species, sufficiently abundant to do a great dea]
in keeping Aphis
in check, but unfortunately they come rather
late and
much harm is often done by the Aphides before they make
their presence
felt. Syrphus
ribesil,
Linn., and
S. grossulariae,
Meig., seem
to be
the
most abundant after the one you send. These three species were sent from widely different localities. First Report on Economic Zoology. 8o The Rosy Rustic
{Hydrcecia micacea) attacking
Potatoes. Amongst
the
numerous
enemies
from
wliich
the
potato
crop
suffered during 1902 were
the
larvae
of
one
of the noctuid moths
known
as
the Eosy
Rustic
{Hydrcecia
micacea). This
attack was
reported to the Board of Agriculture from two localities, namely from
"Winton, ]\Ianchester, and from near Wigan, Lancashire, and w^as
also
observed personally. The two notes sent with these
larviie were as
follows
: (1) Enclosed
are grubs that have attacked and are destroying a large
patch
of potatoes
;
\y\\\ you kindly say what they are and what remedy I
can take. (1) Enclosed
are grubs that have attacked and are destroying a large
patch
of potatoes
;
\y\\\ you kindly say what they are and what remedy I
can take. can take. (2)
I
herewith
send
two
insects
that
I
have
found
amongst my
potatoes. They were looking well to about a week ago. Any information
about them will be thankfully received. (2)
I
herewith
send
two
insects
that
I
have
found
amongst my
potatoes. They were looking well to about a week ago. Any information
about them will be thankfully received. In both
cases the
caterpillars were found
to
be working
in
a
similar way, namely, by tunnelling up the
stalks
of
the
potatoes,
completely hollowing them out and so
killing the haulm. Should
this pest become very numerous
it would
be
a
serious matter,
as
remedies
are
quite
out
of
the
question
save
hand-picking
the
attacked haulm. These
caterpillars
are recorded
as feeding
in
the
stems of equisetums, docks,
valerian, but probably attack a variety
of other plants. The
larva when mature varies from an inch and a quarter to an
inch and a half in length. The back and sides
are
dull purplish-
brown, paler on the
first three segments
and
where
the
segments
join, the sides and venter are of a dull flesh coloirr, the legs pale and
the head yellowish-brown
; on the second segment is a brown semi-
circular plate broadly margined in
front with blackish-brown and a
shiny yellowish-brown patch on the anal segment with a
posterior
border of small dark warts
; on the segments are small dark-ljrown
tubercular warts each with a
fine terminal hair
; the
spiracles
are
deep brown and the prolegs pale with black extremities. Before pupation
the
larva
becomes
a
paler
dull smoky
flesh
colour all over, with a dusky median dorsal line. Beetles on Barley affected with Smut. Specimens
of
barley
affected with
" Smut " and
small
beetles
found with
the fungus
were -received on
26tli
June
(1901) from
]Mr. Neame, of Faversham. Mr. Neame
stated
that he found them
in a field of barley badly affected with smut
; they were of a glistening
black colour, and seemed to be only on the ears affected by smut, and
occun-ed on almost every foul ear that had recently speared. Where
the smut had begun
to blow away he could not
find them. They
seemed
to be eating through the skin
of
the
ears. The following
answer was sent to his letter of enquiry
: The Beetles you send are known as Plialacrvs comisms, Paykull. They
are common and generally distributed throughout the London and Southern
districts, rather common in the Midlands, rarer further North. I do not
know anything
of the economy of the
five
British
species, but they are
certainly not
injurious. Two N. American
species, F. ^Jolihis,
L., and
P. penicillatus,
Say, have been noticed
to feed upon the
spores
of
the
Smut-fungus on wheat. There is no doubt, I think, that those you notice
in your barley are there, as you say, to
feed
off the fungus
attacking the
crop, and that they may therefore be looked upon as beneficial creatures
;
but
at the same time
it
is very probable they carry the spores about with
them and so help spread this serious cereal malady. The Rosy Rustic. 8i Rosy Rustic and Surface Larvce. 83 hour by one specimen
under
observation. The damage caused by
a small number in
a plot of potatoes
will be seen to be very great. The larvre have been reported as early as May 10th in dock. Stainton
says
it feeds on the roots of various Cyperacea. The pupal stage is
found in the ground in an earthen excavation. It is light yellowish-
browTi in
colour, about three-fourths of an inch
long, ending
in an
anal spike and some short stout bristles, pointing backwards, on the
last two segments. The imago appears in August and September, a
few stragglers even
in
October. It
is
particularly found in gardens
and
lanes
and by hedgerows,
also
along
the
borders
of
ditches,
marshes and fens. It
flies
at night and readily comes to light. The
fore wings
are pale brown with
a rosy
tinge,
a
dark
broad patch
below the upper border between the inner line and the elljowed line. The hind wings are whitish-grey with a darker central line. The wing
expanse varies from one and a quarter to one and a half inches. An almost
identical attack
is
recorded
from
America *
by two
species, Hydrcecia nitela and H. nehris, Guen. The former
is known
as
the
Potato
Stalkborer and has
been
recorded
doing
injury
to
potatoes
in Western Maine, the
damage
being
due
to
the
larvse
boring up
the stalk and causing
it
to
wilt. It
also
attacks wheat
and carnations
in Ohio (Webster). It
also injures tomato, spinach,
cauliflower, dahlia, aster, lily, spirwa, salvia, thistle and other plants,
and has been noticed in currant, apple, peach and
blackberry twigs,
and
wheat
and
other
corn. No doubt our species
has
a
similar
varied
diet. I do not know the egg stage of this moth, nor where
the eggs are deposited in this country. Pl{EVENTIOX. All we can do
is to hand-pick the haulm
in gardens and fields
where
it
is seen
to
wilt and then destroy the
larvae
inside
; by
so
doing
a
great deal of damage
will
be
saved. Poultry should
be
turned on potato fields, when the crop has been lifted, to devour the
pupa?, and the men should be instructed
to
kill
all pupae they turn
up in digcrine:. ^00" *
" Some
Insects
injurious
to Vegetable Crops,"
F. H. Chittenden, U.S.
Dept.
Agri., Div. Ent., Bull. 33, 1902. The Rosy Rustic
{Hydrcecia micacea) attacking
Potatoes. The full-fed stage
is reached from the beginning to the end of July. During the whole
of
its life the larva burrows up the haulm and emits a great quantity
of green frass
;
a round exit hole
is made in the stem, the frass being G First Report on Economic Zoology. 82 emitted
tliroiigli
this. Buckler quotes
a
letter from the Hon. T. de
Grey as follows
:—" I
first observed the
larva by pulling up, on the
14th May, a sickly-looking plant of Uquisetum
arvense. It appeared
to be feeding on the root and stem below the surface of the ground, emitted
tliroiigli
this. Buckler quotes
a
letter from the Hon. T. de
Grey as follows
:—" I
first observed the
larva by pulling up, on the
14th May, a sickly-looking plant of Uquisetum
arvense. It appeared
to be feeding on the root and stem below the surface of the ground, Fig. 9. the potato stem-borer {HydrcBcia micacea). a, Imago
;
1) ami d, larva
;
c, pupa
;
e, hole in potato haulm caused by larva. (Natural size). Fig. 9. the potato stem-borer {HydrcBcia micacea). a, Imago
;
1) ami d, larva
;
c, pupa
;
e, hole in potato haulm caused by larva. (Natural size). but when placed
in
a
bottle with a
supply
of
the
food
plant,
it
immediately
entered
a
stem,
and
fed
upon
the
inner substance,
hollowing it completely out, and ejecting the frass at the lower end."
This describes the way it has been observed working in potato haulm. They work with great
rapidity, eight inches being
tunnelled
in an but when placed
in
a
bottle with a
supply
of
the
food
plant,
it
immediately
entered
a
stem,
and
fed
upon
the
inner substance,
hollowing it completely out, and ejecting the frass at the lower end."
This describes the way it has been observed working in potato haulm. They work with great
rapidity, eight inches being
tunnelled
in an Rosy Rustic and Surface Larvce. Surface Larvae attacking Celery and Potatoes. The
Heart
and
Dart
Moth
(A. exclamationis,
Linn.)
larva,
commonly
called
Surface
Larvte
or Cutworms,
were
reported
by
Messrs. Carter and
Co. as
seriously attacking one
of their
clients' *
" Some
Insects
injurious
to Vegetable Crops,"
F. H. Chittenden, U.S. Dept. Agri., Div. Ent., Bull. 33, 1902. G
2 First Report ou Economic Zoology. 84 celery and
potatoes. They have
been very abundant
this year in
many parts
of the country, and have caused
a great deal of harm to
all kinds of roots. Various remedies have been suggested and used with varied results. Soot and lime broadcasted over the
soil and worked in with a prong-
hoe
in an attack on
turnip has been
successful. Bran
soaked
in
Paris green and placed in heaps just under or on
the
soil has been
known
to
destroy them
; hundreds
being
poisoned
l3y
eating
the
arsenic on the bran. Kainit, at the
rate
of 3
cwt. to the
acre, has
also
been
successful. In
regard
to
the
attack
on
potatoes,
it
is
difficult
to do any good, but with the
celeiy,
soot and lime broad-
casted on each side of the rows would prove beneficial. One grower has recommended watering with
paraffin emulsion. If the plants are young this would no doubt be an excellent remedy. {Batliyscia
'wollastoni,
Jans.) Early in June, Mr. Stains, gardener to F, Monins, Esq.,
of Eing-
would, near Dover,
sent some
potatoes badly damaged and
full
of
small Snake Millepedes (vide pp. 15, 32, and 86) {Julus lyulchellus). These often serious pests had undoubtedly done most of the damage. At
the same
time, either by accident
or by intent, there were sent
with the potatoes a few small brown beetles. These were identified
by Mr. Waterhouse as Bathyscia wollastoni. Canon Eowler, in his " British Coleoptera" (Vol. III., p. 71), says
that this species is " found in rotting seed potatoes."
Mr. Stains was
so
informed, and
the
subject
of
these
little
beetles
dropped. But
early
in
July
they
appeared
upon
the
scene
again. I
noticed
some
" Up-to-Date "
potatoes
in my garden with
the haulm badly
damaged
;
there were no
signs
of any larvre to be seen either by day
or at night. My gardener said the damage was due to small "brown
bugs "
in
the
soil, and on
digging up some
tubers
I found them
covered with this small beetle. Eather more than two-thirds
of the
crop proved unsound, some from " rot," but the majority owing to the
]\avages of this beetle, Not only is the sound seed potato eaten, but the tubers themselves. The beetles work first
of
all along the
surface
of the
potato, eating
surface galleries, and then tunnel
little round holes
into
the tubers
these tunnels and channels soon decay and turn brown, and
so the
tuber rots away. An attacked potato cut in two looks
as
if riddled
with fine shot. The seed potatoes seem to
l)o hollowed out
; whether o Beetle,
uent decay I could n
M
St i
t Pigmy Potato Beetle
85 Pigmy Potato Beetle, 85 this
is due
to
the
beetles
or subsequent decay I could not say
for
certain, but I believe from the latter. On July 14th, I wrote
to
this
effect
to Mr, Staines, to which
lie
replied that he had
also come
to
the conclusion that these beetles
were causing much harm. Not
content with damaging
potatoes, they spread into an onion B B Fig. 10.—THE PIGMY POTATO BEETLE (Batliyscia ivoUastcmi, Jans.)
A, Section of damaged potato (c, holes eaten by the Ijeetle, D)
; B, damaged
outer suiface. Fig. {Batliyscia
'wollastoni,
Jans.) 10.—THE PIGMY POTATO BEETLE (Batliyscia ivoUastcmi, Jans.)
A, Section of damaged potato (c, holes eaten by the Ijeetle, D)
; B, damaged
outer suiface. bed and ruined
quite half the
crop. Just
as
in the potato crop, so
with the onion, they work underground. The
rootlets of the onions
were eaten off, so that the plants fell down
;
the beetles also nibbled
at the lower part of the onions and so caused them to split and become
worthless. I
tried them with numerous
other food
plants
experi-
mentally, and found parsnips the only one they would touch
; but
if
a
parsnip, an
onion, and a potato were put
in
the same
box, they bed and ruined
quite half the
crop. Just
as
in the potato crop, so
with the onion, they work underground. The
rootlets of the onions
were eaten off, so that the plants fell down
;
the beetles also nibbled
at the lower part of the onions and so caused them to split and become
worthless. I
tried them with numerous
other food
plants
experi-
mentally, and found parsnips the only one they would touch
; but
if
a
parsnip, an
onion, and a potato were put
in
the same
box, they First Report on Economic Zoology. 86 invariably sought the potato, and when that was gone the onion. I
have not had
sufficient time
to
trace the
life-history
of this beetle,
which can only be looked upon as a serious potato pest in East Kent
and probably elsewhere, its small
size and peculiar habits rendering
it very inconspicuous. From enquiries I have made, it seems
to be
commonly distributed over
that
part
of the
county, and
has
been
connected with the diseased state of potatoes Ijy many local men. The
beetles
(Fig. IOd)
are
very small, never more than
2 mm,
long
;
oval and convex, of a dull, reddish-brown colour, with iine dull
yellowish
hairs
and
the
surface with minute punctures
; the thorax
a
little broader than the elytra, rounded
at the sides and contracted
in front
; the elytra are narrow behind and rounded at the apex
; legs
long and
slender, and the
tibia?, especially the middle
ones, spinose. Canon Fowler records
it from Finchley and Hammersmith
;
all
the
other records are from Kent. {Batliyscia
'wollastoni,
Jans.) They are extremely active, lioth when on the potato and
in the
ground, running with great rapidity and falling from the tuber at the
least shock. Evidently they hibernate
as
adults, for when going
to
press (December) they are still alive. Millepedes attacking Potatoes. Mr. F. Powers
forwarded some
potatoes from
Great Staughton,
badly attacked by small Snake Millepedes {Julus ^pulcliellus) ; but two
other species were present, one Ijelonging
to the genus Jnlus and the
tliird to the genus Polydesmus. These animals breed in the
soil and
often cause considerable
loss
; they are probably primarily attracted
by the decaying seed. Another correspondent, Mr. Staines, writing from Eingwould, near
Dover, says
: I am sending you two
potatoes which
are
attacked
by some
pest,
a
small, thin, worm-like pest, enclosed with the tubers. As you will see, they
attack the
old
set, also the young
tuber. AVhen
it attacks the old
set,
it
entirely
destroys
the
crop,
the
haulm
ceases
to
grow,
and
eventually
disappears. In my
opinion,
it
is
quite
as bad
a
pest
as
PhijtlioiJliora
infestaiis. If
you can
give me any information
regarding
the
pest
I
should be extremely
obliged. Last
year when planting
I
dusted round
each tuber soot and
lime,
with good
results. This year
I
did
not
use
soot and lime, and am very much troubled with
it. Liming the soil is the only treatment of practical use on a large
scale. There is no doubt Mr. Staines's plan of dusting soot and lime
around
each tuber when
set
is a good plan—lime being the great
specific against Millepedes. A plan that has met with success where o Beetle. animals
is given
in th
re Bathyscia wollastoid,
84). Colorado Beetle. 87 gardens are troubled with
these animals
is given
in the Eeport
on
p. 115. The small beetles also sent were Bathyscia wollastoid, an abundant
potato pest in East Kent (vide p. 84). Notes on the Colorado Beetle in England. 88 serve as baits for any fresh beetles that might appear in the spring of
the
following
year. In
spite
of
the
heavy
dressing
of gas
lime,
volunteer
potatoes
and
weeds
germinated
freely,
showing how
uncertain gas lime is in its action. The length of the different stages of the beetle
in England, as far
as my observations
go, are
as
follows
: The egg
stage, ten days
; the
larval
stage, from three weeks
to a month or
five weeks
; and the
pupal stage, from a week
to ten days in the summer. The eggs are
usually laid on the under surface
of the leaves
in
clusters
of from
nine to
forty, but Mr. Craigie has observed the
beetles at Tilbury to
deposit a few on the upper surface as well. They are very conspicuous
elongated oval orange bodies attached to the leaves, after the manner
of those of the Lady-birds. The eggs were found at Tilbury also on
the leaves of the Sow-thistle {Sonchus). There
is some
variation
in
their colour
: some are deep orange, others pale orange, according
to
their age. All the
beetles
I brought away from
Tilbury in
the autumn of
1901 went
to earth by the middle of October and remained under
ground
all the time,
except during a few warm days
in November,
when two appeared on
the
surface
of the
earth. They made their
appearance
above ground
in April, but
did
not
all come up
until
May 4th. They were kept
until May 20th, when they commenced
egg laying. These " check " specimens were then destroyed. At
this date no beetles had made their appearance at Tilbury, so
that the probability is that the
drastic measures taken
last autumn
cleared off all those in the adult stage. According to Eiley, the beetle passes the winter only in the adult
stage, and thus it was hoped that the measures taken had exterminated
the pest in this country. But in May, 1902, beetles commenced to appear again at Tilbury. Those that I saw alive were
certainly not hibernated specimens,
so
easily told by their dingy appearance. The specimens appearing
at
this time were without doubt freshly-hatched
ones, the
elytra being
quite pale compared with those that hibernated. It thus seems that
the Colorado Beetle passes the winter
also in the pupal stage in the
soil. Notes on the Colorado Beetle in England. The appearance of
the Colorado Beetle in England
in 1901 was
not
surprising. Ships from American ports may very
easily bring
over specimens that have flown on board whilst
in harbour on the
other
side. A
single pregnant female woudd be enough
to
start
a
colony. It
is probably in
this way that
tlie colony established
in
1901 in Tilbiuy Docks originated. It has been surmised that the pest
may have come over in American potatoes
;
that is scarcely likely, for
the only American potatoes
that
are sent
to
this country
are seed
potatoes, which are particularly clean. At present there is no infesta-
tion of the Colorado Beetle in Europe, so that
it can only have been
derived from
an American
source. Two
previous
outbreaks
have
been known in Europe, both in Germany. The outbreak
of this beetle in 1901 was reported to the Board of
Agriculture during August. The
beetles were
then very
vigorous,
•eggs and larvtB of all sizes being found. The colony was evidently well established, and looked as if it had
been in existence some
little
time. It was situated
inside Tilbury
Dock on some potatoes on the workmen's allotments
; they extended
for about
fifty yards, and had
materially defoliated the
potatoes
in
some places. The adults did not readily seem
to
take wing, but crawled about
with
great
energy. Later on
in the
year,
those
taken away
for
breeding pm-poses, however, showed a strong disposition to fly
; they
were constantly flying with great
force in the breeding-cage in which
I kept them, and beautiful objects they are, too, when
their
brilliant
rose-coloured under wings are expanded. By keeping some specimens
in warmth, I succeeded in getting through a complete life-cycle by
•October, but those kept out-of-doors did not deposit any eggs
after I
brought them away from Tilbury. The land where this beetle had taken up
its abode was cleared of
all potato haulm, and the haulm burnt with paraffin
at night on the
ground
under
the
superintendence
of
the
Board
of
Agriculture
officials
; the
ground was
also
soaked with
paraffin, ploughed
ten
inches deep, and then dressed with gas lime
at
the
rate
of 60
tons
per acre. Potatoes were planted on and around parts of
tlie area
to First Report on Economic Zoology. Notes on the Colorado Beetle in England. This will account
for
this second outbreak at Tilbury. There
is no doubt
that some
larvce had
already
buried themselves
deep
in the
soil previous
to
the
plot being
treated, and no doubt many
of
these were below the ten inches ploughed up, and
so escaped the
effects of the gas lime and paraffin. Professor Howard informs me that
this
is known
to happen
in
America
as
well
:
Professor Smith having
observed
the
beetle
to 89 Colorado Beetle. pass
the winter
in
the
pupal
stage, which, however,
is apparently
exceptional. This secondary method
of passing the winter makes the insect
more difficult
to cope with when
it makes
its appearance in a new
country, and should be borne in mind in the destruction of any future
colony that is found. The number of beetles that appeared
this
year (1902) was not
large, but
sufficient
to show that they can well survive our winter,
even under such unfavourable circumstances
as existed on the plots
at Tilbuiy. From specimens in the National collection it seems that there are
three closely-related species of Borypliora, namely, D. ^mdccemlineata,
Stal
; D. juiida, Germ. ; and D. melanotJwrax, Stal. The first-named
has
black
legs,
but
otherwise
resembles
the
Colorado
Beetle. D. mclanotliorax has an
entirely dark
thorax, not yellow with black
spots and
central mark, as seen in the Colorado Beetle. D. juncta^
known as the Bogus Colorado Beetle, has two
of the black lines on
the wing cases very closely united, forming almost one broad single
line. There
is
a
fourth
species
in
the
collection, D. multitceniata,
Stal, but there seems to be no difference between
it and D. undecem-
lineata. The only species likely to be confounded with the potato pest
is
D. junda in its larval stage
; but it can easily be told when young by
being much paler than decemlineata, and when adult by having a pale
head instead of a black one
;
its eggs also differ, being wliite instead
of orange. The Pteports sent to the Board of Agriculture on visits paid to the
infested plots are appended. First Report on Colorado Beetle Outbreak at Tilbury. Beport of Secunil
Visit to Tiluarii re Colorado Beetle. Beport of Secunil
Visit to Tiluarii re Colorado Beetle. On September 17th,
11)01, I
visited Tilbury with Mr. Craigie to make
further examination of the allotments upon v.hich the Colorado Beetle had
been found breeding. No
signs
of any fresh
beetles had been
noticed by the foreman who
had
charge
of
the
plot
of
land. None
could
be found under
Ijoards,
sacking,
etc.,
places
where
they
might
hibernate. It
is
extremely
improbable that any could
live
in the ground covered with the gas lime
in the way
it has been treated. No signs of any damage could be detected
on the potatoes, not yet dug, on the railway bordering the infested patches
in the docks. There is a possibility that a few stray indi\'iduals might live in amongst
the rough grasses
neor
the
plots, ^\hicll
although
treated with gas
lime,
still
offer many tempting
spots
for
wintering
where
the
lime
has
not
fallen. It would be
well to plant a few batches
of
early potatoes about, as
traps
for
any
that may have
escaped
;
this
would
probably
stop any
stragglers in the spring from straying aw-ay. The specimens taken away at ]ny first ^isit have now all gone to earth. On placing them amongst fresh leaves and on the soil, even in the sun, they
refuse to remain above ground. These will be kept as checks, and as soon
as they commence to appear in the spring from the soil, information will
be sent to the Board, when a sharp look-out may be kept at Tilbury. For
the present nothing further can be done. First Report on Colorado Beetle Outbreak at Tilbury. First Report on Colorado Beetle Outbreak at Tilbury. The potato plots in the allotments iu Tilbury Dockyard were visited on
Aiigust 22nd, I'JOl. g
The Colorado Beetle {Dorijphora 10-Uneata) was found
to be present. The
beetles were not
at
all
numerous, not more than two dozen being
observed. They were, however, very active, and breeding was going
on. Larvas-
n all stages were found and three batches of ova. They were, however, very active, and breeding was going
on. Larvas-
n all stages were found and three batches of ova. The beetles seemed to be limited to about fifty yards of
the allotments,
the end
nearest
the
station
being quite free from the
pest. A single
specimen was also found on
the Nightshade. None were detected in the
rough herbage surrounding the
plots, a likely place to harbour hibernators
during the winter. g
The beetles showed great vitality, but the damage, although noticeable First Report on Economic Zoology. 90 on the crop, was not us great as one \vonld
expect. Amongst tlie potatoes
mentioned were several
of
tlie
large Ladybird {Cocclnella
septempuiiciata),
Avbich were feeding on
the eggs
of
the
beetle. It wonld
be advisable to
have all surronnding
plots examined, especially those on the other side of
the high fence separating the dockyard from
the
railway. As the beetles
occur on poppies and tomatoes both these plants should be searched. RecomrmndMlions. 1. Constant supervision and hand-picking adults and ova on infested
plot in the docks for one month. 2. Very careful examination of the potato
plots along the railway line
just outside the dock property. 3. Clearing
off the rough herbage
in
close proximity to the
infested
plot. (It would be aS well not to burn this near
; smoke very often makes
insects fly when they otherwise would be sedentary.) i. An examination
of
all potato
fields and plots within a three-mile
radius of the dockyard plot. This had best be made twice
; once as soon
as possible, and again about the 21st of June. A
visit ought
to be paid now as
well
as
later,
because some
of the
beetles may have flown away last year and
hibernated, and
like those kept
at Wye, have appeared during
the
early part of May and commenced to
breed. If this has happened the larvae should be quite large. 5. It would be well
to have the
leaflet on
this pest sent to all potato
growers on both sides of the river to distribute to their men. Colorado Beetle. Colorado Beetle. 91 It is probable
that they may go on appearing
for another two weeks,
bnt
I
think
it
well
to have a watch kept
over
the
plot
for
at
least a
month. That a brood may appear over some time we may judge from the
fact that eggs and
larvse
in
all
stages were found
last August at the same
time, although
it
is probable
that the
first brood would hatch out about
the same time. Bcport on the Colorado Beetle at Tilhury (1902). I have
visited
the
plot
of
land
in
Tilbury Dock
upon which the
Oolorado
Beetle
bred
last
summer,
and
found
that
the
beetles
were
appearing in small numbers and
that they had already commenced to lay
their eggs (June 2nd). As
stated by Mr. Brown, the beetles were coming
out of the ground that had been treated with gas lime and paraffin during
August, 11)01. The
beetles seen
by me were
quite fresh specimens and
presented
a
nuich brighter shiny appearance than those
I
kept
alive during the winter
at Wye
; one specimen found during my visit on June 2nd
anjis certainly
immature. It thus seems that these
beetles
that are now appearing have
passed the winter
in
the pupal
stage. The larv^ had no doubt gone to
earth before the land was treated
last autumn. A number may well have
gone deeper than
10
inches,* so
that they would escape
the action of the
gas lime and paraffin, and thus the appearance of the
beetles this year can
be accounted for. * The land was ouly ploughed to this depth. * The land was ouly ploughed to this depth. No.
6.
Chrysomela marginalis, Duft. Insects sent as Colorado Beetles. A number
of insects were
sent
to the Board of Agriculture
as
Colorado Beetles from different localities
;
it is a matter of interest to
note
the
great
variety
of
creatiu-es
sent—none
being
the
pest
in
question. The specimens are as follows
: q
p
No. 1. No. 2. No. 3. ) The Cockchafer {Melolontha vulgaris, Fabr.). No. 4. No. 5. )
No. 6. Chrysomela marginalis, Duft. No. 7. Bibio hortulanus, Linn, (a fly). No. 8. Larva3 of one of the Clirysomelidm (Colorado Beetle family)
of the genus Timarcha. No,
9. Bmying Beetle {NecroplioriLs sp.). No. 10, The Cockchafer {M. vulgaris, Eabr.). No. 1. No. 2. No. 3. ) The Cockchafer {Melolontha vulgaris, Fabr.). No. 4. No. 5. ) No. 6. Chrysomela marginalis, Duft. No. 8. Larva3 of one of the Clirysomelidm (Colorado Beetle family)
of the genus Timarcha. No,
9. Bmying Beetle {NecroplioriLs sp.). No. 10, The Cockchafer {M. vulgaris, Eabr.). 92 First Report on Economic Zoology. No. G
is never in
sufficient numbers to do much harm. No. 8
feeds
chiefly on
rank
grasses and low
herbs, and
does no harm. No. 9 is beneficial, the beetles and their larvee acting as scavengers. No. 7 is
to some extent injurious
in the
larval stage
; the larvce
occur in large
masses
in
the
soil and attack the
roots
of various
plants,
especially
grass
;
gTeat
numbers
have
appeared
in
some
districts this spring. ^
'
-1
9' > Melolontha vulgaris, Fabr. ^
'
-1
9' > Melolontha vulgaris, Fabr. No. 13. Pyrochroa serraticornis, Scop. No. 13. Pyrochroa serraticornis, Scop. No. 13. Pyrochroa serraticornis, Scop. This latter is a very common
beetle, which passes
its larval and
pupal stages in rotten oak, birch, beech, willow, and other wood. Neither beetle nor larva are in the least destructive. No. 14. Telephorus rnsticus, Fall. No. 15. TclcpJwms hicolor, F. No. 16. Melolontha vv.lga.ris, Fabr. Nos. 14 and 15
are popularly known
as " Soldiers and Sailors."
They are most
voracious, the females even devouring
theu' mates
;
the larvce are
also carnivorous, feeding on insect grubs, earthworms,
slugs, etc., so that these beetles should be protected. Nos. 14 and 15
are popularly known
as " Soldiers and Sailors."
They are most
voracious, the females even devouring
theu' mates
;
the larvce are
also carnivorous, feeding on insect grubs, earthworms,
slugs, etc., so that these beetles should be protected. No. 17. Clytus aridis, Linn. No. 18. Melolontha vulgaris, Fabr. No. 19. Crioceris asparagi, Linn. No. 20. Melolontha vulgaris, Fabr. No. 20. Melolontha vulgaris, Fabr. No. 19 is the Asparagus Beetle. No. 17 is of little importance. No. 17 is of little importance. Nos. 18 and 20 are the large Cockchafer. Nos. 18 and 20 are the large Cockchafer. No. 21. Bihio hortnlanus, Linn. No. 21. Bihio hortnlanus, Linn. No. 22. Larvae of Chryomelid Beetle (Timarcha). No. 23. Steropus mandidus, Linn. No. 24
)
.^
j^P
V Melolontha vidgaris, Fabr. No. 26. Pterostichiis vulgaris, Linn. No. 26. Pterostichiis vulgaris, Linn. No. 23 attacks mangolds and strawberries, eating the fruit
of the
latter plant. No,
24
also
attacks
strawberry
; both
are somewhat
abundant
this season. No,
24
also
attacks
strawberry
; both
are somewhat
abundant
this season. No. 25.-)
No. 30. \ The Eosy Eustic {Uydrcecia micacea). No. 31.j No. 25.-)
No. 30. \ The Eosy Eustic {Uydrcecia micacea). No. 30. \ The Eosy Eustic {Uydrcecia micacea). No. 30. \ The Eosy Eustic {Uydrcecia micacea). No No. 30. \ The Eosy Eustic {Uydrcecia micacea). No. 31.j The pupa sent to the Board of Agriculture, numbered
32, is the
pupa of the 7-spotted Lady-bird (Coccinella septempuoictata). The pupa sent to the Board of Agriculture, numbered
32, is the
pupa of the 7-spotted Lady-bird (Coccinella septempuoictata). Cabbage Aphis. 93 Reported Colorado Beetle at Hockley. Reported Colorado Beetle at Hockley. Some pupae sent from Hockley were those of the 7-spotted Lady-
bii'd (C. scptempiLnctata). Supposed Colorado Beetle at South Benfleet. Pupae sent from South
Benfleet
as Colorado
beetles were those
of
the
7-spotted Lady-buxl (C. septempunetata). Suspected Colm^ado Beetle at Northfleet. Pupse also sent from Northfleet were those of the 7-spotted Lady-
bird (C. septemjmnetata), and
not any
stage
of the Colorado Beetle,
Nothing was found in the tube resembling Pig. 2 of the pamphlet,
but the pupae are those figured at No. 7. There
was
also
sent
a
small
adult
Lady-bu'd
{Hippodamia
variegata). No. 13. Pyrochroa serraticornis, Scop. There are no
records
of any
of
the
true
Lady-bii'ds, except
a
single
species
of Suhcoccinella, S. vigintiquatuorpunctata, Linn., doing
any harm
to
foliage in
this
country, but many
of the large genus
Epilachnct, which does not occur here, do considerable harm, and are
all
herbivorous and
not
carnivorous. It
is
extremely improbable
that
tlie
potato
leaves
sent
by Mr. Craigie, which appear
to
be
devoured
to some
considerable
extent, have been
attacked by
the
Coccinellids
sent. It
is much
more
likely
that "Surface
Larv?e,"
which are nocturnal feeders, were the cause of the damage, or slugs. At the same
time,
it
is
of course not impossible for carnivorous
insects to develop herbivorous habits. Search should be made on the
potatoes
for Plant Lice,
the normal Lady-bu'd food
;
if none
occur,
then there would be more reason to think that the larval Coccinellids
had become herbivorous. The Cabbage Aphis on Turnips. A
correspondent forwarded
in
September
some
turnip
leaves
seriously damaged by aphides from the Isle of Thanet. The leaves
were attacked by the Cabbage Aphis {Apliis hrassiccc, Linn.)
Nothing
can, of course, be done
as late as this
;
as a rule the Ichneumon flies
parasitise them
in
enormous
numbers
in
September. It
is
not
known
for certain how they pass the winter—but probably in
the
egg-stage on wild Cruciferae. This
species
does not usually attack First Report on Economic Zoology. 94 turnips or any other root-crop leaves, but Curtis
evidently observed
it on
the stems of turnip
leaves. Its normal food
plants
are the
various Brassicce, upon which
it may occur in great numbers, causing
large crinkled
folds and swellings on the
leaves, which turn white. The Aphides are covered with a white mealy coat. Unless one has a
field Strawsoniser one can do nothing in such
attacks. Their Life-history and Treatment. Several
enquiries
have
been
received
during 1902 concerning
Leather-Jackets. The so-called Leather-Jackets, or the
larvae
of the Daddy Long-
legs,
or
Crane
Flies,
that do most harm
to
crops, belong to
five
species, namely, the common Crane Fly {Tipula olortcea);
the Marsh
Crane
Fly
{Tiimla
j^oJndosa)
;
the
Striped-abdomen
Crane
Fly
(T. lateralis, Meig.)
;
the Yellow-Spotted Crane
Fly
{Pacliyrrliina
maculosa), and an allied species, P. quadrifaria. Some years it is one
species
that does most harm, in other years
another, or
all may be
equally abundant. During the year 1902 the Yellow-Spotted Crane
Fly (P. maculosa) was most abundant
generally. The
larvae
of
all
species work in a very similar way, the grubs feeding upon roots of all
kinds of plants, often working into the interior of large roots just below
the surface of the ground. In such plants as the dahlias, carnations
and hops, they often cannot be detected, as they work so far into the
roots. They not only attack plants below ground, but they frequently
appear on the surface, and have been noticed to eat through straw-
berry runners. Their appearance on the surface
is chiefly at night. I have frequently noticed those
of oleracea and maculosa feeding in
large numbers above ground on damp summer nights. Eitzema Bos
has not only observed the larvae of maculosa feeding above ground at
night, but
also " by day in dark, damp weather," and watched them
at
work on
the growing
field
crop. This habit
of coming above
gi'ound
at night
to
feed
is one we must pay especial attention
to
from an economic point
of view. All these
larvae
are particularly
prevalent in grass land and
clover
lay, where they find
congenial
surroundings amongst
the tangled
gTowth
of
roots and
in
turnip
fields
; but at the same time we get them in
rich, clean garden
soil,
causing havoc amongst lettuce, cabbage, and tender flowering plants. During
the
past
season
(1902)
the
larvte
of
P. macidosa
were
observed working into the stems of cornflowers just below and above Injurioits
TipulidcB. 95 ground, and caused complete destruction
of beds
of quite large size. It
is
mainly
in
undisturbed ground
that
these
insects
propagate,
especially when
there
is
moisture,
as in damp meadows, marshes,
and amongst the vegetation along dykes and
ditches. The smaller
larViT. of F. Their Life-history and Treatment. maculosa occur most abundantly on
light
soils, but not
by any means
entirely, for I have seen them in swarms during the
past
year
on
clay
land. They
occur
in
hilly
districts
just
as
abundantly as in low-lying marshy land, in light and heavy soil
; in
fact, they have as wide a distribution as the common Crane Fly. The larvae of all these and other injurious species live throughout
the winter, feeding all the
time, except when tlie ground
is frozen
;
they then pass deeper into the earth to escape the cold. Some seem
to reach maturity sooner than others
of each
species,
for
it
is not
infrequent to find adults of some of the species occurring over several
months, but the main brood occurs
about
the
same
time
;
others
have two
or more broods in the
year. Grass land and root
crops
perhaps
suffer more
than anything
else
from
the
ravages
of
the
Leather-Jackets. The
following
instances may
here
be
recorded. In
1813, according
to Kirby and Spence
(" Introduction
to Ento-
mology "),
hundreds
of
acres
of
grass
land
were
destroyed by
Leather-Jackets. In
1842
the
marsh
lands
by
the
side
of
the
Thames
in
the Isle
of Grain were so completely destroyed by these
grul)s that the ground was bare. This occurred again in 1894 in the
same
area. The
larvae, of
course, differ
in
certain
features
in each
species. In general form they are cylindrical, without any feet, with
a distinct horny head,
retractile,
i.e., it can
easily be drawn into the
succeeding segments
; the posterior end
is truncated and ends
in
a
number
of fleshy
projections,
so-called
papillae, which
vary in
the
different
species. There
are
two
respiratory
orifices
on
the
last
segment. The mandibles are dentate and work transversely, not upon
one another, but upon
two other
fixed
pieces. They
are not only
found
living in
roots, but
also in rotting wood and even in water,
both
salt and
fresh. The pupae
of
these
insects can
easily be
dis-
tinguished by their having two horn-like
projections from the head
;
the segments of the abdomen are encircled more or less with spines,
especially beneath, and, like most of the nematocera, are naked, that
is they are not enshrouded
in
a
puparial
case. Their Life-history and Treatment. This
stage
in
the
root-feeding Tiimlidai
is always found in the ground where the larvae
have been feeding, generally at some little distance from the suiface. Just
Ijefore the imago
is ready
to emerge they wriggle partly out
of the ground,
the abdominal
spines
being used
for
this purpose
usually about half the pupa
projects above the
level
of the
earth. First Report on Economic Zoology. 96 The imagines then escape. It is a very common sight to see hundreds
of these empty pupal cases sticking up amongst a few square feet of
pasture. They
are
especially
noticeable, projecting from
the edges
of lawns along gravel paths. These
insects do
endless
mischief to
lawns,
but
never
to
the same
extent
that they do
to permanent
pasture, because the mowing and rolling, especially if carried on late
into the autumn, kills so many of the
adults, and destroys the eggs,
besides compressing the ground
so firmly that the Leather-Jackets
can move but slowly from root to root. Very frequently the damage done to grass land by their larvre is
attributed
to
other
causes. Miss
Ormerod
gives
the
following
instance
:—" On May 24th Mr. W,
Gray,
^vl•iting from
Langholm,
Dumfriesshire,
N.B.,
sent me
some
quite
young
caterpillars
of
the Antler Moth of various
sizes, from very small up
to as much
as
a
third
or half-grown. He mentioned
at
the same
time
the
injured appearance
of the
grass, but that on searching for the cater-
pillars there seemed very little
sign
of them, which he ascribed
to
their being
still so small that they escaped observation. However,
about a month later the true cause of the damage was found."
Tlie
maggots
proved
to
be
the
larvae
of
P. maculosa
(Eeport
XIX.,
p. 33, 1896). p
The
five
chief injurious
species may have
their
characteristics
summarised as follows
: I. The Common Ceane-Fly. (Tipida oleracea). {Tipula paludosa). This
species
is nearly as common as the former, wliich it
closely
resembles. It appears, however, a little
later as a rule, and may be
at once distinguished by the wings
of the female being shorter than
the body and by the absence of the pale streak under the costa in the
female;
this
pale
streak,
however,
occurs
in
the
male,
but
the
(genitalia
differ
from
those
of
oleracca. The
legs
are
also much
stouter than in
oleracca. The body, especially in the female, is
of a
general ferruginous colour, with the
dorsal
stripe weakly developed. The palpi are also stouter than
in the common crane-fly. Its larvse
feed in similar situations to the former. I am not acquainted with
its structural differences. I. The Common Ceane-Fly.
(Tipida oleracea). This species (Fig. 11, i) is widely distributed over Great Britain, its
larvae and those of the next species being the common forms of large
Leather Jackets so destructive to
all
crops. The adults appear from
]\Iay
to
September,
the
majority being
seen
during August
and
September, but they may occur even
into October
in considerable
niimbers. They can stand a
fair amount of frost, for I have seen
them
alive
after
the night
temperature
has been
as low as
28'' F. The
adult
is
silvery-grey
;
the
thorax
striped
;
the
metathorax
silvery-white
;
the
abdomen
slaty-grey
;
the
segments
becoming
testaceous
towards
their
edges,
and
there
is
a
dark
lateral
line
between
the upper part and the testaceous
sides
; the apex
is
also
testaceous. The long,
slender legs
are testaceous
;
the
tarsi dark
brown. The wings
are longer than
the body,
greyish;
the
custa
brown, and sharply contrasted from the rest
of the wing, and hcncath
it there is a greyish, li^npid streak in both $ and 9
. The larvae when full grown reach an inch in length and about the The larvae when full grown reach an inch in length and about the Injiirioits
TipiilidcB. 97 thickness
of a
goose-quill. The
skin
is
quite tough when they are
mature, much
wrinkled, and
of an earthy
color. The
blunt
tail- 5
il
Fig. 11.—INJURIOUS
TiintUdee or daddy-long-legs. 1, Tipula
oleracea
;
2, T. lateralis
;
3, Pachijrliina moculosa;
i and
5, pupa and larva
of
T. ukracea
; 6 and 7, pupa and larva of P. macidosa
;
8, eggs of
T. oleracea
;
9, of maculosa
;
10, basal cell and veins near, in Fachyrhina
;
11, in Tiimla. •end
is
furnished
with
four
large
tubercles
on the edge
with two 5 il 5 5
Fig. 11.—INJURIOUS
TiintUdee or daddy-long-legs. 1, Tipula
oleracea
;
2, T. lateralis
;
3, Pachijrliina moculosa;
i and
5, pupa and larva
of
T. ukracea
; 6 and 7, pupa and larva of P. macidosa
;
8, eggs of
T. oleracea
;
9, of maculosa
;
10, basal cell and veins near, in Fachyrhina
;
11, in Tiimla. •end
is
furnished
with
four
large
tubercles
on the edge
with two
below, and in the centre the two respiratory openings. H H First Report on Economic Zoology. 98 They pupate in July, August and September. I. The Common Ceane-Fly.
(Tipida oleracea). The pupa is about
as long as the larva, but not
quite so thick, and in colour varies from
dirty brown
to brown
; the ventral spines
are
large, and
there
are
small ones on the dorsal parts of the segments
;
the tail-end is spiny
and acuminate. dirty brown
to brown
; the ventral spines
are
large, and
there
are
small ones on the dorsal parts of the segments
;
the tail-end is spiny
and acuminate. The larvpe of this
large Tipulid feed on all manner of roots, but
are especially prevalent in grass land. {Tipnlct lateralis, Meigen.) are especially fond of damp, wet, muddy earth. The
other two
recorded
injurious
species
belong
to
the
genus
Pachyrhimi.s of Macquart. The members
of this genus can be
told
by their more fragile form and black and yellow colouring, and they
have
the
three veins from
the
discal
cell, generally
starting from
separate bases (Fig. 11, 10). {Tipnlct lateralis, Meigen.) This
is
a
very abundant
species which
sometimes
appears
in
swarms. I
have
frequently had
the
larvte
sent me
as damaging
grass lands from different parts of England. I have noticed
it
to be
particularly abundant along the grassy roadsides in Huntingdonshire
some
years, notably in 1890 and
1900, when great numbers
of the
maggots were attacking the grass in neighbouring fields. Grass roots
seem
to be
the main food of the larva?. There
are no records of
it
attacking garden produce or other
crops, but it doubtless does so. It
occurs
in the adult
stage in June and July and again in September. In the latter month I found
the flies swarming
in the fields around
Sidmouth in 1889. The thorax of this species (Fig. 11, 2) has three brown stripes and
is margined with deep brown
; the middle line
is broadest
anteriorly
and has
a
dark
central
line
in
front. The abdomen has chestnut-
brown
side
lines, and
eacii segment has
the
posterior border with a lujitriGiis
Tipulidce. 99 fme pale line. The legs are reddish-brown, the tips of the joints dark
brown. The wings are tinged with brown and there is an oblique pale
mark by the stigma. The
cross-veins are clouded with dark brown
and the marginal
cell yellowish-brown. Its length varies from half
to two-thirds of an inch. The larva varies from three quarters of an
inch
to
nearly an
inch
in length
;
it
is thick
skinned,
of a dirty
brownish yellow hue, often with a coating
of earth when
it assumes
a brownish appearance, and has three dark stripes running down tlie
body on the back interrupted by the segments
;
there are a few dark
short
hairs
;
the anal end with four short thick
papillae
above,
all
much the same
length, but
the two middle ones closer together and
a
little smaller than the outer pair, and two short, coarse ones on the
lower edge. The pupa is nearly an inch in length, of a dirty whitish
colour at first, becoming
blackish-brown. On the ventral
surface of
the fifth
to the eighth segments
is an unequal sized transverse row
of bristles near each
posterior border
;
there
are
also spines on the
front parts of the ventral segments
;
the
last segment
is surrounded
by ten spines, four above, four belo-w', and two on each side. The larva? (PachyrJiina maculosa, Meigen.) This
is
a most abundant
species in Great
Britain in fields, road-
sides, and
especially in gardens. It appears
in June and July and
again
in September. After
the two
large
Crane
Flies
this
is the
most harmful species, some years
it being far more destructive
in
its
larval stage than they are. Its
life-history was first worked out by
the greatest economic entomologist England has seen—John Curtis. During the season
of 1902
it appeared in enormous numbers
in
some
districts,
such
as Kent and
Huntingdonshire, and
has
been
reported in great abundance elsewhere. I also found it swarming in
parts of Devonshire in 1888. Curtis speaks
of
it
as swarming on the sea coast, and mentions
" seeing myriads on sand banks
in
the
Isle
of Portland, also
at the
back of the Isle of Wight, and at Lowestoft in Suffolk." H
2 First Report on Economic Zoology. loo Most of the small " leather-jackets "
infesting gardens are of this
species; they
especially attack
lettuce,
peas,
young
brassiere,
and
garden flowering plants. There
are
two
broods during
the
year, and
in
some
seasons
possibly
a
third. Curtis
recoi'ds them
as
early
as
]\Iay. I have
taken it in numbers as late as August. The
adult
(Fig. 11,
5)
is
yellow, the abdomen
having a broad
interrupted dorsal line
;
the head has a dark triangular patch behind
;
the thorax three black stripes, the lateral pair curved outwards at the
front end
; and the sides (pleurae) before the halteres blackish-brown
on three sides. The wings are transparent with a pale brown stigma. The thin
delicate
legs
are
testaceous, dusky towards the
tips. In
length this species varies from a little under to half an inch. The eggs of P. maculosa are oval and jet black. The larvse when
mature
are
never
more
than
three-quarters
of an inch
long. In
colour they are earthy and the skin
is wiinkled, but not tough as in
the Tipula:. They are cylindrical, somewhat attenuated at each end
the alimentary canal shows through the
skin, above and below, as a
broad dark stripe. Each segment has a transverse row of four
stiff
bristles, the inner ones
of each row the
shorter
;
laterally are short,
.stiff, black hairs. V. The Allied Spotted Crane-Fly.
{Pachyrhina quadrifaria, Meigen.) This is a closely related
species
to the preceding, and like
it
is
generally
distributed over England, but does not seem
to occur
in
such swarms, nor do its larvse seem to occasion
as much harm. It,
however, has been sent
to me from various parts
of
Surrey and
I
have
observed
its
larvte
in
great
numbers
ravaging
flower
and
vegetable plants
at Kingston-on-Thames in 1884 and 1886
;
in the
latter
year
it was
especially
abundant
in
the
south
of
England. The adult appears in June and July. The female is yellow, the abdomen with a black dorsal stripe
; the
head with a triangular black
spot
behind. The
thorax
has
three
broad black
stripes, the
lateral pair including two yellow spots, and
the metathorax
has three
black
stripes. In the
$
the abdominal
stripe is inteiTupted on the anterior border of each segment, in the 9
the dorsal stripe is dilated on the hind border of each segment. The
wing is transparent, with the stigma brown, and the hind
cross-vein
and the last piece
of the vein below
it infuscated. Legs testaceous
tips
of the femora and
tibise
black, and
the
tarsi
dusky. Length,
half an inch. The
deep
brown
stigma
will
at
once
separate
it
from
the
preceding species. The larva
is seldom more than half an inch
long, of a greyish-
yellow colour, with
thick
skin, and very like
that
of
P. maculosa
foui' dorsal papillae, the two
inner ones much
shorter than the two
outer ones
; the two ventral papillse short, also brown stripes beneath
the stigmas. The pupa is about as long as the larva, brownish-yellow,
with sharply indented
segments
;
two
short,
thin,
rather
spatulate
cephalic
horns
;
each
segment mth
six
or
seven
dorsal
spines
ventral surfaces with five teeth-like spines. Injurious
Tipulid<x. loi (PachyrJiina maculosa, Meigen.) They can
at once be
told from the large leather-
jackets when
the
latter are immature,
i.e., about the
size
of matm^e
maculosa grubs, by the anal processes
;
in
this
species the truncated
tail has two hooks or papillee, and two short ones between them, with
two blunt tubercles below and two
fleshy protuberances capable
of
dilatation
and
contraction
;
there
are
also
two
central
spii\acles
between each stigma and
the
ventral
papillpe
a
transverse row
of
three small dark brown
spots. They reach maturity in the spring
and pupate
in
the
soil. The pupae are brown
to golden brown in
colour, slightly narrower than the
larvae, and have the two
straight
cephalic horns
; the abdominal segments have each a row of minute
spines
above
and
six
large
ones
beneath, and
on
either
side an
elevated spiny line, the penultimate segment has six long spines and
two small ones, and there is a large conical process at the
tail with a
shorter one beneath
it. Curtis
describes them
as not only eating
roots, but also eating off trusses of the strawberry flowers close to the
crown. He
also found them
in May
at the
roots
of
lilacs
and
amongst the roots of grass
; they also destroyed carrots, raspberry and
strawberry
roots,
lettuces and various
flowers. Miss
Ormerod,
as
previously noted, gives records of its damage in the Scottish uplands,
where
its working was mistaken for that of the larva of the Antler
Moth (Charceas (jraminis). Injurious
Tipulid<x. Natural Enemies of Tipulid/E. The Tipulidse are preyed upon by
a number of natural enemies
wliich, however, are not
sufficiently potent to stop them doing much
harm and causing great
loss both to the
agriculturist and horticul-
turist. The
subject
of
natural enemies
is
one
to which
special
attention should be paid, but
it
is quite useless
to expect very great
benefit
to
accrue
from
any
except
the
birds. People who
are
acquainted more with the laboratory than the field talk of the use of
parasitic
hymenoptera
{IchnPAimonidce
and
Chalcididce)
and
of First Report on Economic Zoology. 102 predaceous
insects
as
if they would do all that
is required to check
an
insect
pest. In
a
few
cases
predaceous
insects
have
done
enormous good, under abnormal
conditions, but in a
state
of nature
they
never
appear
in
sufficient numbers
to make any
impression
until the pest
in question has increased
to such an extent
that the
harm
is
all done. The
introduction
of new
predaceous
forms
is,
however, always worth
trying,
as now and
again good
has
resulted,
as
seen in the
case
of
the
Icerya
scale
and
Vedalia
cardinalis. When
people commence
to
talk
of
stopping spraying
for Aphides
or scales because the beneficial parasites are killed also, it is sufficient
evidence
they know
little
of
fruit-growing
or
the
fruit
grower's
troubles. In
the
Tipulidte we
find
scarcely
any record
of
insects
that
destroy
either
larvfe
or
adults. Curtis
mentions
Ichneumons
as
attacking the larvne, but says no more. Although
I have bred large
niimbei's from
different parts of Britain
I have never come across a
parasite, with the exception of a single 9 Tachina, sp. (?) that appeared
in a cage of T. olcracea. Numerous birds, however, feed very largely on
these insects, not only upon the larvie, but also upon the adult flies. Amongst those birds which
are great " leather-jacket " destroyers
are the following
: the Eook, the Starling, the Peewit, various Gulls,
the Pheasant, and, to a lesser extent, all
the
Ttirdidce, or Thrushes,
and Blackbirds. Enormous numbers of these
larvae are devoured in
autumn,
winter,
and
spring
by
the
three
former
birds,
and
the
pheasant's crop may frequently be found full of them. The decrease
ia
the number
of
Peewits
has
been
marked by
a
corresponding
increase in this
pest as well
as
in
the Wire-worm. PREVENTIVE AND KeMEDIAL MEASURES. Although there is no known remedy
for " leather-jackets " in the
field
there
is much we
can
do
to
lessen
the amount
of harm
caused by them and
to prevent
their undue
increase. In
garden
cultivation, on the other hand, we can destroy them even in
the soil,
if it is considered worth tlie while. In the tirst
place, we can do some good in preventing egg
laying. This we may do
in three ways;
firstly, by keeping down
all long
herbage during the autumn, long rank growths
at the headlands and
along
hedgerows
;
secondly,
by
bush-harrowing
pasture
land
or
heavily rolling the same when the swarms of flies
are noticed in the
fields,
hundreds
will
thus
be killed and
so prevented from laying
their eggs, and the eggs in many cases themselves will be destroyed
;
tlmxlly, late mowing and
rolling of lawns, croquet grounds, etc., will
not only kill large numbers
of the
adults, but eggs
as well. There
is no
doubt
that
for
preference the flies will go
to damp
areas
to
deposit their eggs, and thus
drainage will do good. This
has been
carried out on many occasions and has always been attended with
good
results. Pasture land
and
clover lay
should
be
broken
up
when
possible
early
in
the autumn, so
that
the
flies
can
find no
shelter amongst which
to
lay
rheir
eggs. The
land may
first
be
dressed with gas lime, spread over it
at once and allowed
to remain
on the
surface for a few days. The smell would deter the flies from
laying their eggs and would probably affect any small larvae present. The effect
of gas lime
is very variable, however, and
it
is doubtful
from recent experience if
it has much
effect upon large subterranean
insects. The old plan of
" paring and burning " the
stubble
or grass on
breaking up pasture
is perhaps the best method
of clearing out this
and other ground
pests. It
of course has
its disadvantages and
is
now seldom
practised, but there
is not the
least doubt that it is the
only way to lessen these pests in the soil. Eolling with a cross-kill or Cambridge ring roller does some good
by compressing the
soil, and
so
preventing
the
" leather-jackets
''
from free movement in the ground. This
is
especially advantageous
where they are attacking wheat
or
barley. Injurious
Tipitlidce. Injurious
Tipitlidce. 103 Natural Enemies of Tipulid/E. All
these birds
should be encouraged, not only because of their destroying " leather-
jackets," but other pests as well. The adults
are devoured by the Spotted and other Fly-catchers,
by the Swallow, and even by the Sparrow. The Rook
also devours
large numbers as they are ovipositing in the
fields. Poultry also do
much good,
for they feed whenever
opportunity occurs upon both
larvre and
adults. These birds
all do good because they
are always
present and are constantly feeding. They are often attracted in greater
numbers when hosts of insects appear, but even when normally present,
by devouring these and other pests
before many of them have done
harm, they do
inestimable good, whilst on the
other hand
parasitic
and predaceous insects come too late. Moles
also
feed
off leather-jackets, and do
far more good than
harm
even
in
pasture
land,
where
their
" heaps "
cause
some
inconvenience. PREVENTIVE AND KeMEDIAL MEASURES. In dealing
with
their
life-history it was pointed out that the grubs come
to the surface at
night
; a heavy ring-roller, of course, would kill any number of their
larvse
if
passed over them, and could not
fail
to
do much
good, First Report
oii
Ecoiioinic Zoology. 104 especially where
as
in grass
land we have absolutely no means
of
controlling them. The extra cost of night work
would
be
amply
repaid. It
is recorded by Miss Ormerod " that
the
grubs may be
collected by a top dressing of rape-cake and the
roller
passed over
the ground in the morning with good results "
;
this does not always
seem to have the desired effect, however. Traps may be employed in gardens in the form of heaps of turf,
partly buried in the soil
; the
flies lay their eggs there and the
larvae
seem
to
be
attracted
to
it and can
then
be
collected and
killed. Eotting turf-manure and leaf-mould heaps should be kept free from
surface weeds and top dressed with gas lime, or else they will form
breeding grounds
for these Tipulidte, and the larvae will be
carried
to field and garden, and so contaminate the
soil. Grass-borders in
gardens are favourite breeding grounds, and from
these
the
larvae
spread to the bedding plants. In garden borders and beds they can
be easily destroyed by injecting bisulphide of carbon into the
soil at
the rate of half an ounce to every square yard. This should be done
in
late autumn
or early spring. When crops
are attacked various
stimulating manures should be employed
; they do not kill the larvae,
but they hasten on the growth of the plant and repair some of the
damage that the grubs have done. Nitrate of soda has the greatest
effect upon leather-jackets, but
is not permanent
in
its
action. It,
however, causes a cachectic condition in these pests, from which they
do not recover for some days, if its application at the rate of 2^ cwt. to
the acre
is followed by rain. Miss Ormerod records an experiment
in wliich
at the
rate
of 2
cwt. to the acre they had not recovered
from
its
ill
effects
after
eight
days. The
effect
of
this
artificial
manure on insects is so extremely variable that one must not rely on
it too much. Section IL Animals Injurious to Horticulture. Animals Injurious to Horticulture. Julidae destroying Plants in the Gardens of
Downton Castle, Ludlow. In answer
to a communication received from Mr. C. Boughton
Knight,
of Downton
Castle,
regarding the damage and annoyance
caused by Myriapoda, the following report was sent
: The
Millepedes that are damaging
strawberry
and
other
plants
in
the
gardens
at Downton
Castle
are
the small Snake Millepede {Jidus
pulchellus). These animals
live both on healthy and diseased plants. The eggs are
laid
in the ground
; the young JuU have but few legs
at
first. I have
found them breeding at all times of the year, but especially in spring and
early summer. Jidi/s 'pvlchellus is particnlarly prevalent in damp localities
and where decaying vegetation is left about. Heaps of leaf mould harbour
it especially. Lilies are
particularly subjected
to the ravages of this pest,
but all roots seem to be liable to its attack. I have made one
or two experiments
Avith
the ones
sent me
from
Ludlow, and I find that poisoned bait
is very satisfactory. I simply dipped
the pieces of mangold and potato in a strong solution
of Paris green and
covered the
baits with a cabbage
leaf. This morning
all the Millepedes
were dead, those not so treated all alive and well. I think
I shonld adopt
this plan
to
clear them out
in preference
to
any other. The baits should be larger than the pieces sent with the pests
and should be left
to soak in
the Paris green
for an hour
;
of course the
poison,
which
is
soluble
in
water, must be
kept
stirred up every now
and then. Put
the
baits down wet and cover
over with a green
leaf, the
best
time to start them would be at night (dusk). Heaps
of
leaf mould,
etc., should have a good dressing
of gas lime
mixed with them if the creatures are observed there. There were
also a few Polydesmiis complanatus with the Julus
;
they
also are easily poisoned. There were
also a few Polydesmiis complanatus with the Julus
;
they
also are easily poisoned. PREVENTIVE AND KeMEDIAL MEASURES. At the same time
it must be borne in mind that if
it
has
not
the
desired
effect
on
the grub,
it
is
still of
value
as a
stimulant
to
the
plant. Hand
and
horse
hoeing
have
also
been
recommended and largely followed, but
the good
done
is scarcely
sufficient for the outlay. PVoodlice in Gardens. 105 *
" Some
Insects inju'ions
to the Violet, Rose, and
otiier Oraaiiiental Plants,"
Bull. 27 (N.S.), U.S. Dep. of Agriculture, 1901, p. 47. Woodlice in Gardens. In answer to a
letter sent by Sir William Thiselton-Dyer, from
Mr. Thomas A. Lance, of Sydenham, Surrey, the following reply was
sent concerning Woodlice
: The
scientific name of the woodlouse most commonly found in Great
Britain
is PorcelUo
scaler, Linn. There
are two
other common
species First Report on Economic Zoology, io6 tlitit do
harm,
viz.,
Onisci/s
asclli's,
Linn., and ArmadiUidium
vulgare,
Lat. Oniscus aseUus
is omnivorous, but does nuich harm
in liot-liouses
and to soft wall-fruit
;
it also eats away at strawberry roots. This species
rolls
itself up
into a ball and can be
told from the ArmadiUidium and
Porcellio by having eight-jointed
antennse, the two
latter having
seven-
jointed ones. j
ArmadllUdium vidgare
is larger and
of a uniform
slaty blue and rolls
itself up very readily. p
y
y
Porcellio
scaler
is
brown, much variegated
in
colour,
with a rough
shell and two long spines behind. Probably they have been
spread
in
the manure from
the heap you
refer to. You
might
cover
the
manure
heap
with
lime—gas lime
(hot)
in
preference—but it must be left on the heap for some four weeks before it is
put on the land. J should put a layer an inch thick of gas lime over the
heap and let it stand for some time. Woodlice may
easily be trapped along
borders by putting here and
there pots filled with moss and horse dung. They can be collected in the
day-time and so destroyed. Many plants are harmed by these pests
;
as a rule the harder the leaf
the more the plant escapes. There
are some twenty
species
of Woodlice found
in
Great
Britain. These
land
isopoda
are included
in twelve
genera. They may mostly
be found under moss, decaying wood, and leaves, both out-of-doors and in
greenhouses. Some few, such as Ligia
ocecmica, Linnaeus, and Fhiloscia
Gouchii, Kinahan, seem to be partial to the neighbourhood of the sea. t This
is
described
under
the name
Eriophyes
violae,
n.
s]).
(Sitzung
der
matiiematiscii-naturwissenschaftlicheu
Classe
vom
11.
Dec.
l0O2,
Kaiserlichc
Akademie der Wissenschaften in Wien). A New Phytoptid Disease
in Violas. Quite a new disease in violas has Ijeen reported by Mr. Charles J. Gleed,
of Cliveden Gardens, Maidenhead. The specimens sent had
most
of the leaves curled
tightly over
at each
side and were quite
hopelessly deformed. Mr. Gleed wrote that he
" thought
it was the cold weather
; but
the attack is not
general, two or three
plants here and
there, about
30 per cent,
of the
plants and both young
stuff struck
this
spring,
and
old
plants
off which
cuttings have
been
taken,
are
attacked
indiscriminately." At
first
sight
one would
say
the damage was due
to
Biplosis
violicola—the Violet Gall-Midge described by Mr. Chittenden * and
excellently figured—but an examination soon revealed the real cause
of
the
disease. There
were
found
in
all
the
leaves
examined
a
number
of short, thick
green phytopti which seemed
especially
to
congregate towards the apex of the leaves. As many as fifty of this
large species were counted in one leaf. It is larger than the Currant The Narcissus Fly. 107 Gall Mite, and cau easily be seen with a hand lens. Specimens were
sent
to Dr. Nalepa, who informs me
it
is a new species which he
is
going to describe.t Information was sent to Mr. Gleed to destroy the plants that were
attacked and all cuttings taken from them, and to burn the earth in
which they were growing. If allowed
to
spread,
this mite would probably form a
serious
source of loss to nurserymen. {Mcrodon
equcstris, Fabr.) {Mcrodon
equcstris, Fabr.) A correspondent, Mr. T. J. Leney, sent the
larvpe of
the
Nar-
cissus
Fly, from
Chertsey, with
the
following
note
:
" They
play
havoc with the narcissus bulbs and are evidently the maggot of some
fly. They commence boring from the
base
of
the
bulb
upwards,
eating out the
centre. I cannot quite understand
their
beginning
operations at the base of the bulb
; one would have thought the eggs
would be deposited at the top and the maggots work down the bulb,
whereas the point of entrance
is in nearly all cases as shown by the
dot in the drawing." The larvas were of the dipterous genus Merodon, several of which
are known
to
feed upon
the bulbs of the narcissus, etc., in Europe. One
species only
is
so
far
recorded
from
Great
Britain, namely,
Merodon equcstris, Fabr., according to Mr. Verrall, but Walker in
his
work
on
" British
Diptera "
also
gives
Merodon
clavipes, Meigen,
probably in error. There are three varieties of Merodon equestris, viz.,
var. narcissi,
F.,
var. vcdidus,
Meig.,
and
var. transvcrscdis,
Meig. These
three were
at one time treated
as
distinct
species. Which
variety the larvre sent belong to it is not possible yet to say. This Narcissus Fly has frequently attacked the bulbs
in
Corn-
wall, and I have had it reported to me from Ham, in Surrey. The fly appears in May, and may be seen flying over daffodils and
other similar plants. The
fly resembles
to some
extent
a
bee
in
form
;
it
is three-fourths
of an inch long
;
the body is deep bluish-
black
with
transverse
bands
of
golden
yellow;
the
wings
grey,
fringed with dull yellow
;
legs black, short and stout. The female probably lays
her eggs near or upon the bulb. Tlie
larvcne, however, always seem
to enter from
the
lower
part
of
the First Report on Economic Zoology. io8 bulb
as you
describe. Those
I have kept attacked as many as ten
bulbs
before becoming mature. When one bulb
is destroyed they
crawl through the soil and enter the next one at its base. The grubs
seem
to
reach
maturity during November, but
some
kept
under
observation lived until January. When
full fed they form a cell in
the earth near the
last bulb attacked and
line
this
cell with silk,
forming a perceptible cocoon. The Marguerite Fly and
its Destruction. Specimens of Marguerite leaves tunnelled and generally damaged
were received from Great Staughton,
near
St. Neots, on
the 4th
of
June. The
following
reply
was
sent
in answer
to a request
for
information in regard to destroying the culprits
: The white daisies that are attacked
are
being tunnelled by the
little
grubs of a small fly, the Marguerite Fly {Napomyza lateralis, Fall). The
only thing one can do
is
to
pick off
all the
diseased
leaves, and
if
they
are very bad destroy (burn) the whole
plant. There
is no remedy, and
if
left alone they go on spreading very rapidly. When the plants are young
spraying with
paraflBn emulsion wiU
keep the fly away, but when
it has
once laid its eggs on the leaf you can do nothing for
it. {Mcrodon
equcstris, Fabr.) The puparium
is dark brown, oval
in
form, and has two projecting processes in front. It
is probable
that
this pest
is
constantly being imported from
the Continent. It
is very desirable
to
lift
all bulbs
in a bed
that
is
invaded
before October ends, even if it is not the year for their removal. All
bulbs should be examined and any showing decay destroyed, or
the
liy will go on increasing and may do endless
harm. Where
beds
have been invaded and the bulbs
lifted, the ground should be deeply
dug and the top spit buried so
as
to prevent the flies emerging next
year. Mr. Leney informs me
that on
lifting
the
bulbs
to
have
them examined and
to
kill
the
larvae no more than two full-grown
larvae occurred in each bulb, but more frequently only one in a bulb
when, however, the larvae are in a younger stage and about half the
size
of those sent
(fully mature) he invariably found from seven
to
eight in a single bulb. ^to'- The
Carrot
Fly
[Psila
rosce,
Fabr.)
and
Aphides
on
Carrots. The
Carrot
Fly
{Psila
roscc) was
very^
destructive
during
the
summer of 1902. One
correspondent, Mr. Hammond, WTiting from
Canterbury,
states
as
follows
:
" I
have
since looked
at my neigh- Insects
in Orchid Houses. 109 bour's
carrot-bed
; every carrot has been
ruined
;
his bed
is utterly
ruined."
Together with the work
of
the
Carrot Fly in
this
neigh-
bourhood was a bad attack
of Aphis. " People about here," writes
Mr. Hammond, " are
complaining that their
carrots
are very much
infested with Aphides
at
the
roots."
This was
early
in
October. By the
20th
Mr. Hammond
found
that they had
nearly
all
dis-
appeared. They
are
to be
found
generally on
the crown
of the
carrot
; they do not cause the
cracks, but
shelter
in
them. These
Aphides
were
too
shrivelled
to identify when they
arrived
; they
were probably Schizoneura fodiens. Gatton Pabk, Surbey,
Btn July, 1902, Gatton Pabk, Surbey,
Btn July, 1902, Btn Jul
To Pkofessor E, Ray Lankester, M.A., etc.,
Natural History Museum, South Kensington, S.W. My Dear
Sir,—I am a collector of orchids and somewhat largely
interested
in their hybridization, but our efforts are materially
interfered
with by a little fly or its grub (specimens of which
I
enclose), and whose
ravages we
have found no
means
of
preventing. It
is no uncommon
thing for them to clear off the whole of the contents of a pot of seed
as
soon as it is sown and germinates. We have made the following observa-
tions in regard to it
: They seem to frequent and thrive most where they have damp moss to
dwell
in, which unfortunately
is an essential
feature of
successful
orchid
growing. Being often found on plants that have no seed on them, they of course
have other food, but apparently they have largely
increased
in my houses,
which, however, may be due rather to the amount of moist moss about than
to
the amount of
orchid
seed which
they have
to
feed
on. They are
equally troublesome in what we know as the " Cool House " (50° to 00° F.)
as in the hot ones (65° to 85° F,). They devour the seed immediately
it
commences
to germinate, and
if not devoured
in
this
stage they attack
the
small bulblet
as
soon
as
it
is formed, eating
it from the base and
leaving the shell only. They are harmless
to
plants after the early stages
and so are not troublesome to orchid growers generally. Careful
search
has
failed
to discover
a
grub. Hence
it
is assumed
(possibly erroneously) that it is the fly which does the mischief. Further,
the winged one (? male)
is rarely found on the
pots, which
leads
to
the
assumption that it is the wingless one which does the mischief. If we are
wi'ong
in the assumption that the male only has wings our observation
would lead to the suggestion that the fly
is harmful only before
it develops
its wings. The body of
the wingless one on the pots
is much larger than
the
one
found
with wings. When
the
seed
is sown on
a
flat
surface
without harbour we are not
troubled
; but
directly the
seed
is removed,
which
it has to be soon after germination, it becomes liable to attack. Gatton Pabk, Surbey,
Btn July, 1902, First Repoi't on Economic Zoology. no Various
methods
have
been
unsuccessfully
tried,
particularly
the
following
: Various
methods
have
been
unsuccessfully
tried,
particularly
the
following
: Fumigation, either with tobacco or a compound known as " X.L, All
(this
is supposed
to
contain
nicotine, camphor, methylated
spirits, etc.),
but neither has any material
eifect. Quassia affects them only for a short
time, as they leave the pots and return in a few days. The pots have been
placed under water for hours, but upon being taken out the flies are equally
as lively as before. Fly-paper and strings similarly covered are of no avail. y p p
g
y
(N.B.—The houses are regularly fumigated for pests, in general about
every ten days.) I
should be
very
glad
to know
if
there
is any method
of
ridding
ourselves
of
the
pest by
destruction
or
of
making
it
harmless
to
the
germinating seed and
bulblet by
driving
it from
the
pot or otherwise. Any information
as
to
its known habits might
help ns
to work
out
its
destruction
if no remedy
is known. It
is of
course
important
that any
remedy shall be harmless
to
the
orchid
seedlings
themselves. They are
extremely delicate and porous and have
to be kept in a constant state of
moisture. I must apologise for having troubled you with so long a letter, but the
ravages of this insect are most annoying and often rob us of
results which
have promised
to be of
great
interest
in the horticultural
world, and
I
should feel much indebted
if you could let me know of a remedy or of any
one who would
be
likely
to
advise me
should
you
personally
be
not
acquainted w'ith one. I beg to remain, etc.,
(Signed) I beg to remain, etc.,
(Signed) I beg to remain, etc., Jeremiah Colman. Report on Insects in Orchid Houses. The
insects sent by Mr. Colman,
of
Gatton
Park,
Surrey, causing
harm to orchids, are in too broken a condition to identify accurately. The
small
flies are SciarincR and belong to the genus Zygonema, of which only
one species occurs in Britain
Z. sciarina (Meigen), found in summer and
autumn
in underwoods and moss—but without seeing fresh and
perfect
specimens
it
is not
possible to be certain of the species. The life-history
is not known, but probably they breed
in the damp moss. The larvs do
not seem
to have been observed
; they would
possibly be
in
the form of
small white footless grubs. The
flies can do no harm—it would be
the
larva3—but
there
is no
douljt
that the damage
is done to
the
orchids by the wingless creatures
sent at
the same
time. There
is no connection
between
the
Avingless
insects and the
Sciarina3. The wingless fonns are Collembola, or Spring-
tails. Some of these are
certainly very
injurious, but
little
is known
of
them, however. The young of these Spring-tails resemble very closely the
adult, and
live and grow in similar situations and under similar conditions
and are injurious during the Avhole of
their existence. Preparations have
been
made
of
this
Collembola,
and
attempts
will
be
made
at
its
identification. The majority of
species
live under damp moss and
stones and
are no
doubt encouraged by
the methods
necessary
in
orchid
cultivation. All
that can be suggested
is
that Mr. Colman experiments on
a small
scale Insects
in Orchid Houses. 1 1 with some common
orchids and hydrocyanic acid
gas. This gas can be
used
for such delicate plants
as maidenhair fern without injury and
is
fatal to
all forms
of animal
life, but
its
effect on orchids has not been
observed. If there
is much moisture on the plants
this gas
loses much
of
its potency, and the
air during fumigation should be dry. Its effects
in an orchid house might not therefore be
as
successful
as under other
circumstances. It
is
certainly worth
trying, however, as fumigation with
tobacco, etc., would have
little
effect on these creatures that
are causing
the annoyance. Great care should be exercised in the use of hydrocyanic acid gas, as
it
is a most dangerous poison to man. Should
Mr. Colman
think
it
advisable
to
experiment
with
this
insecticide, information as to procedure can be sent him. Report on Insects in Orchid Houses. There does not seem to be any other way in which these
pests can be
eradicated under the conditions necessary for orchid cultivation. Fred. V. Theobald. Gatton Park, Sueeey,
2Wi July, 1902. To Peofessor Laxkestee,
British Museum (Nat. Hist.)
Cromwell Road, S.W. To Peofessor Laxkestee,
British Museum (Nat. Hist.)
Cromwell Road, S.W. Apologising for troubling you, etc.,
(Signed)
Jeremiah Jeremiah Colman. To Peofessor Laxkestee,
British Museum (Nat. Hist.)
Cromwell Road, S.W. Dear
Sie,—In
further
reply
to
the
interesting
report
of
Mr. Theobald of
the
21st,
I have had an opportunity of
a
careful
discussion
of the points raised with my gardener. I
understand
the
report
to
throw
considerable
doubt
upon
the
probability
of
the
larvae
of
the
Sciarin^
being
harmful. Under
the
circumstances, and
as
it seems certain that the Coliembola or Spring-tails
are, I suggest that we ignore the former, especially as we have not been able
to observe them and have no actual evidence of their causing mischief. There will be no difficulty in adopting the suggestion that the
effect
of hydrocyanic acid gas upon orchids shall be ascertained by experiments. It
is believed, however, that, used in moderation,
it will not be harmful
to
them. It is certainly unfortunate
that its effect
is
likely to be minimised
by
moisture. The moisture on the moss, etc., can be reduced, but
it
is
such an
essential
feature
in
the
early
stages of
orchid growing
that
it
would have
to
be done
with
great care and
not for any length of
time. Should
it be the young Spring-tails which are most largely responsible for
the
mischief,
material
relief
from
these
pests might
be
secured by an
application of the gas
before the seed
is sown or before the germinating
plants are transferred thereto and when the compcst
is
in
a
perfectly dry
state. A good deal depends upon
their habits, but
I am
writing on
the
assumption
that
the young may not develop very quickly or be
able
to
reach
the
pots
before
the
plants
are
sufficiently strong
to
take care of
themselves. It seems difficult to place the pots in such a position as to be
out of reach of the adult Spring-tails, but we will gladly experiment if any
suggestion can be made. We have now placed some of the pots on a zinc
tray on
stands, which seems
to have minimised
the
mischief somewhat. Although the more orthodox way of
raising the seed seems to be to sow it First Report on Economic Zoology. 1 1 2 upon the moss surrounding growing
orchids, it
is quite possible
to
raise
it and
transfer
it to
very small
pots, so
that
if we have
the means of
preventing the depredations of these
Spring-tails over a small area a great
deal would be accomplished. Section
III. Animals Injurious to Forestry. Goat Moth Larvce attacl^iug IVitlows. 113 The times
to
t'uini^iite
for
]\Ieiily Ww^
are
(a)
before
the
vines
bloom
;
(J) after the crop has heeu feathered. The house shouhl
l)e well A'entilated
for
at
least oue hour
after
fumigation
before
anyone
should
go
into
it,
the
-windows
being
arranged so as to open from the outside, and also the door. The cost comes to about
4r/. per 1,000 cul)ic feet. Tins treatment
has been found not to damage CAen maiden-haix ferns
if carried out
properly, and there is no danger
if proper precautions are taken. It
is not
advisaltle
to
leave the treatment
to
ignorant
people, as
the
fumes and tlie cyanide are of course deadly poisons. Fumigation under Glass for Mealy Bug and
other Pests. Frequent enquiries have been made as to the use of hydrocyanic
acid gas under glass
for the destruction
of Mealy Bug. This
treat-
ment will be found to far surpass the old methods of fumigating with
tobacco and various patent compounds. The method of fumigating with hydrocyanic acid gas (HON) for
Mealy Bug, Scale, etc., under glass,
is as follows
: For every 1,000 cubic feet of space use 5 ozs. sulphuric acid, 8 ozs. water, 3 ozs. cyanide of potassium. The water should be put into a jar and then the acid added to it
remember to always add the acid
to the water, and not the water
to
the
acid
;
the cyanide should be
in small lumps and wrapped up in
blotting-paper
; the cyanide is then dropped into the jar of water and
acid and the fumes allowed
to generate
for an hour. It, of course,
lias to be done quickly and with
care, owing to the poisonous fumes
being so deadly
to
all forms
of animal
life. The cyanide should be
dropped
into the acid and water from outside the house
;
this can
easily be done by putting the jar close to the door or window, so that
it can be shut as soon as the packet
of cyanide touches the mixture. When wrapped
in
blotting-paper, some
seconds
elapse
before
the
fumes generate. Jf the glass-house is more than 10,000 cubic feet another jar will
be required, and for every additional 10,000 feet. The foliage of all plants to be treated should be
as nearly dry as
possible. The temperature never more than 60° Fahr. 50° Fahr. is the best
temperature. At heat over 50° Fahr. there is a
risk
of harming the
foliage. Do
not
fumigate
in
a
strong
light,
as
foliage may
then
be
damaged
; fumigate always after sunset. Do not fumigate vines when in bloom
or just before the grapes
have comnaenced to ripen. Goat Moth Larvce attacl^iug IVitlows. Insects on Osiers and Willows. In answer
to
a
request
for names
of insects
obser^'ed by Mr. Marsh, of Milford
School, near Godalming, who gives instruction in
Basket-A\'ork, and who
is growing the
different
^'arieties
of Willows
and
Osiers with
a view
of comparing them, and
also
finding
out
something
about
their
culture
and what
insects
affect
them,
the
following reply was sent
: As far as one can say from yom" descriptions
of the insects
attacking
your willows and osiers, they are as follows
:— (1) The
Minor
Shoulder
Knot
Moth
{Epunda
viminaUs). The
moth
appears
in
July
and
August,
the
lar'\^a
in May
;
the
pupa
is
subterranean. All
the
larvae
of
this genus
live exposed and
extended
along the stems of plants. (2) The
larva)
of Syrphida3 or Hover
Flies
; they are not injurious^
but beneficial, beiug Aphis feeders. (2) The
larva)
of Syrphida3 or Hover
Flies
; they are not injurious^
but beneficial, beiug Aphis feeders. (3) A green Aphis, undoubtedly Siphocorym
caprea,
Fabricius. It
is
found on
all Avillows and occurs from April to July. It
is fairly common
round Guildford, Godalming and that part of Surrey. (3) A green Aphis, undoubtedly Siphocorym
caprea,
Fabricius. It
is
found on
all Avillows and occurs from April to July. It
is fairly common
round Guildford, Godalming and that part of Surrey. g
p
y
(4) This
Aphis
is
called
Melanocanthus
salicis,
Linn. It
is
especially found on
SalLr viminaUs. The wingless forms appear in April,,
the winged females from the end of .June throughout July. It
is recorded
from your district (Guildford) and
I have found
it in abundance on osiers
at Wye
;
it is also recorded from Kentish Town. (4) This
Aphis
is
called
Melanocanthus
salicis,
Linn. It
is
especially found on
SalLr viminaUs. The wingless forms appear in April,,
the winged females from the end of .June throughout July. It
is recorded
from your district (Guildford) and
I have found
it in abundance on osiers
at Wye
;
it is also recorded from Kentish Town. y
(5)
This Aphis
is
Ghaitophorus salicivorus. Walker. It varies much
in hue. They are often seen brick red in colour. (5)
This Aphis
is
Ghaitophorus salicivorus. Walker. It varies much
in hue. They are often seen brick red in colour. (5)
This Aphis
is
Ghaitophorus salicivorus. Walker. It varies much
in hue. They are often seen brick red in colour. Insects on Osiers and Willows. The osier has a great number of insect
pests, especially amongst
the sawflies, cecids or gall midges, moths and beetles. A list of the more important is being prepared. A list of the more important is being prepared. Goat Moth Larvae attacking Willows. Mr. "W. S. Mockett,
of Eamsgate,
wrote
in Septeml)er regarding
the damage to willows l)y the lar\'je of the Coat ]\I(jth. Several other
^correspondents have
also applied
for information both
in regard to
their life-history and ways of destroying the
larvae. Ash, oak, elm,
as
well
as
fruit
trees, are
attacked by
these large
larvie, and they
frequently kill the trees outright. If there
are
not many Coat Moth
larvas
in
a
tree
it
is
quite
possible
to destroy
tliem. This may
l)e done
in several ways
; the
old plan was to insert a wire into the oi)ening
of the txmnel
to
tind
out which way the
tunnel
goes, and
if downwards
use
a
fluid, if
upwards a gas. The
best
fluid
is
paraffin
emulsion,
with
a
little
Paris-green
injected l)y means
of a
S3ainge, the nozzle being forced into the liole
and surrounded by clay until the injection is over. If a gas or fume
is used, sulphur
acts well
; use bee-bellows and blow the fumes
in,
fixing the nozzle as before with clay. About June, smear the trunk of the tree with cow-dung and clay,
mixed with paraffin, as
far up as holes are found
;
this
pre^•ents egg-
laying. By far the best x^lau has recently been found in the use
of
cyanide of potassium. Place a small piece
of stick cyanide in each
hole and then
close up with
clay. The fumes soon
kill
tlie
larva^. within theu" tunnels. I First Repoi't on Economic Zoology. 114 (Schizoneura lanuginosa, Hartig., and Lachnus viminaUs, Fonsc.) (Schizoneura lanuginosa, Hartig., and Lachnus viminaUs, Fonsc.) Specimens
of Aphides
attacking elm and willow were received
in
October,
1901, from
Miss
J, Bmioughs
Norgate, from
Enfield. One, a large gall on the elm, proved
to be the work of an Aphis
of
the same genus as the White Woolly Aphis or American Blight. It
is known
as Schizoneura
lanuginosa. The Willow Aphides Lachnus
viminaUs,
Fonsc. The
correspondent
stated
that
her
attention
was
called
to
this aphis by the number of wasps hovering over a
lilac bush beneath
the willows. They Avere feeding
off the gummy
honey-dew. The
large masses
of aphides were then discovered on
the
willow
in
their
typical
position. William
Curtis noticed
that Insects on Elm and Willow. 1 1 wasps
feed readily
off
the honey-dew
excreted by
this
species
of
aphis, and also that bees
totally disregarded
it. The flow of honey-
dew produced by this species is very copious and does much damage
to the trees and those beneath. It is not
at all unusual for willows
and osiers to be killed outright by
it. The effect of the punctures of these plant
lice
is to leave distinct
brown
scars
in
stripes. This
plant
louse
is
also known
as
the
Aphis
salUjna, Walker, other synonyms being Aphis
salicis,
Curtis,
Aphis
viminalis,
Boyer
de
Fonscolombe,
and
Lachnus viminalis. Passerine. The wingless
viviparous female
is dark yellowish-bro^^Ti
to
greyish-bro\^^l
; the antennne red
at the
base,
black
at the
tips,
and there are two dark spots on the thorax. The abdomen
is much
rounded and in
the
centre
is
a
curious
horn-like
projection
; the
cornicles are large and short and there
are five to
six rows of large
black spots on
it. The legs are deep brown, rather long and hauy. In length they are about 0*16
of an inch. The pupa
is much
like
the
larva, but rather longer and with bright brown wing cases, and
the dorsal tubercle is very large. The winged female is quite a large insect, 4 to 5 mm. in length, of
a dull brown colour with darker marks, the abdomen being spotted
with
black, one large spot placed centrally
;
this spot apparently is
the representative
of the tubercle seen in the wingless female
; the
short
cornicles
are almost conical. (Schizoneura lanuginosa, Hartig., and Lachnus viminaUs, Fonsc.) The long wings always seem to
be carried horizontally when the insect is at rest
; the stigma is long,
nan'ow and black
; the insertion and
cubitus
orange-yellow. The
legs
are
long,
the
tibiae yellowish-red, the
two-jointed
tarsi
deep
brown. As this is certainly a very harmful species
steps should be taken
to destroy them by washing the willows with paraffin emulsion. This
species
is very common
in some
districts
on willows and
osiers. They congregate in masses often half a foot in length and an
inch or more wide
; they are usually grouped side by side with
their
heads pointing downwards. When disturbed these sedentary insects
l)ecome most
active, yet do not leave their abode
; they throw their
long hind legs up and wave them about
in an
erratic manner, \^dth
the probable intention of frightening off the enemy, especially hymen
opterous
parasites. The
effect
of this
species on
the
trees
is very
strange. Some osiers observed this year were killed by them, whilst
others close to only presented a yellow-leafed appearance
; some shed
their leaves, others recovered in a few weeks. Cameron records a case
where
this Lachnus swarmed in
such numbers
at
Carshalton that
trees
thirty
to
forty
feet high had
been
killed by
their poisonous I
2 First Report on Economic Zoology. ii6 influence. It
is
in
osier
cultivation
that
it proves most
danj^jerous
and
it
should
be
destroyed when
noticed
l)y
hand-picking
or
spraying. Pissodes notatus,
Fabr., ravaging Austrian Pines. Damage to Austrian I'ines
l)y the J>anded Pine "\Vee\'il {FUsodes
iiotatas, Fabr.) has been reported by Mr. E. Hyne and
others during
the past year. According to the reports of Continental foresters, PU'^odes notatus
almost exclusively follows the
I'ine Weevil {Hijlohius ahletts, Fabr.). [t is usually found on
trees rendered unliealthy by the Hylohius and \ w r
Fig. 13. The Banded Pine Weevi
( Fissdile.t iiotatu.t). Fig. 12. LaiTii (i) and \t\\\)A (h) of
I'issoiles iwUittts. Fig. 13. The Banded Pine Weevi
( Fissdile.t iiotatu.t). Fig. 12. LaiTii (i) and \t\\\)A (h) of
I'issoiles iwUittts. finishes the damage begun by that
l)eetle. Fissodes notatus occurs in
all manner of places, in wood split
for fuel, in young li^ing stems, in
pine
cones
and
in
the
bark
at the base
of
old
trees. The
chief
damage
it does
is where
it attacks young unhealthy trees. Planted
pines
suffer
more
than
those
self-sown,
(1)
liecause
tlie planting
often throws them back, (2) on account of the crowding in the nurseries
which makes the young trees sickly. The Fissodes chiefly feeds then
on trees attacked l)y the JTi/lohius and those grown on unkindly soil and
thus more or less
uuliealtliy. If the supply of unhealthy trees fails
then these beetles will attack soimd ones. The
beetle (Fig. 13)
is about one-third
of an
incli long and of a
red<lis]i-brown cohjur, irregularly covered with
bright hairs; the pro-
thorax has eight yellowisli spots
; tlie elytra with two broad pale bands
running
transversely across them. The beetles
a];)pear in
April and The
beetle (Fig. 13)
is about one-third
of an
incli long and of a
red<lis]i-brown cohjur, irregularly covered with
bright hairs; the pro-
thorax has eight yellowisli spots
; tlie elytra with two broad pale bands
running
transversely across them. The beetles
a];)pear in
April and The Banded Pine
M^eeznl. 1 1 May and af;ain in August and September. There seems
to
be one
brood only in the year. May and af;ain in August and September. There seems
to
be one
brood only in the year. The female
beetle lays her eggs
singly, dail}^ or
at
inter\als
of
a fe^v days, egg-laying
lasting
ovei- a period
of several weeks
uj)
to
two
months. The
eggs
are
generally
laid just above the root up
to
as much
as
six
feet
above
the
gr(jund. Pissodes notatus,
Fabr., ravaging Austrian Pines. Egg-
laying commences early in April and
in
]\Iay and may occur again in the autumn,
l)ut
usually
the
females
oviposit
in the
spring. This
l)eetle prefers four to eight
year
old
plants,
l)ut may
attack
those
of much
greater
age. The
larva^
eat
their way between
the
wood
aiul
the
bark, forming
slightly winding passages
Fig. 14. which
inci'ease
in
size
as
the
larvte
Pine cone damasied by I'issodex
notatug. glow. \V hen
mature
they
construct
o\al
filnous cocoons composed of wood filjres
in wliich they pupate. The
larvie. also
live inside pine
cones, whicli they
turn
yellowish-
grey. As many as three larva^ may occur in a single cone. Attacked
plants may be
tohl by small drops
of turpentine on the bark and by
the premature deatli
of
the
needles. The
winter
is passed
in the
l)eetle
stage and
also
in the
lar\al and pupal
stages. The
l)eetles
hibernate
in
the
chinks
of
the
l)ark,
as
near
the
inner bark
as
possible,
mostly where
the
root
and
trunk
join,
generally
abo^-e
ground, but sometimes below. Fig. 14. Pine cone damasied by I'issodex
notatug. Fig. 14. Pine cone damasied by I'issodex
notatug. Peevention and
Ti;eatment. As there
is no doul)t that
the Hylohius
is often followed by this
Pissodes, and that tlie
latter does not often occur without the former,
steps
.should be taken
to destroy the Hylohius if
it occurs
; then the
Fissodes will cease to increase. The causes of Hylohius attack are (1) leaving old stumps
in
the
ground and dead
felled
timljer
al)out
;
(2)
the
presence
of
sickly
trees from either (a) bad
planting,
(]j) unkindly soil, or
(c) gi'owing
the trees too close together. Destruction of all diseased timljer (roots and all) when larva,' and
pupa'
are
in
the
tree
in ^May and June should always
jje carefully
attended to. Laying newly cut stems of ])iue in open parts in April and May First Report on Economic Zoology. ii8 forms an excellent trap
;
in
a few
hours, says KoUar, they will be
found covered with
beetles, particularly so when
the stem
(of
each
trap) has been pressed into the earth. These decoys must not be
laid
too
late and must
all
be burnt
before the brood escapes. This plan has frequently been known
to
clear a forest of Hylobius pest. Billets of imbarked fire-wood
laid about
will attract the beetles
to lay their eggs. These should be destroyed from the end of June
to the middle of July. Smearing the lower parts
of the trunks with a mixture of mud
and lime early in April would probably check egg-laying or perhaps
it would be better still carried out in March. Young
trees containing Pissodes
larvae should be pulled up and
burned in June and July. All
cones
attacked
should
be
collected
and burned
; they may
easily be
told by
the exuding
turpentine. Wood-peckers
(Ficadce)
should be encouraged. Tkeatmext. It is most important that
all gall-bearing trees in young spruce
plantations should be
felled and the
galled
boughs
biu'nt
in
the
summer. In fresh planted areas the trees should be gone over in the
summer and the galls carefully picked off and burnt. They are always most abundant where the trees
are too thickly
planted and on cold
clay
soils
; both
thick
growth
and
clay soil
should be avoided for spruce plantations. GROUP
F. Animals which concern Man as being injurious to his worked-
up Products of Art and Industry, such as (A) his Buildings
and larger Constructions and Habitations,
(B) Furniture,
Books, Drapery and Clothing, (C) Food and Stores. Earwigs causing Annoyance. 119 (Chermes
ahietis, Linn.) Deformed growths on Spruce were sent by Mr. J. Saunders,
of
49, Eothesay Eoad, Luton. These proved to be caused by the Spruce
Gall Aphis {Chermes ahietis, Linn.). These
galls
are
at
first bright
green and rosy and shaped
like a small pine-cone. The " mother
Chermes is oval, wingless, and woolly, green and purple in hue with
blackish
legs. This form
is found
in
the
spring
and
inserts
her
proboscis into the tissue of the plant just below a bud. This causes
the irritation which commences the diseased growth. The female lays her eggs amongst a woolly secretion on the gall
the young
larvse coming from the same stick their proboscides into
the gall which still further swells and grows up more or less around
each
larva. The larvje are really enclosed by the unnatural swollen
leaves
of
the
bud
overlapping
them. Later
these
galls
harden,
become
brown,
the
chambers
split
open, and
the
Chermes make
their
exit. These
soon
turn
to
pup?e, and
then
yellowish-green
winged females, which fly from
spruce
to spruce and deposit about
twenty eggs
each. These eggs give
rise
to
larvce which grow
into
the
" mother-queen " in
the
spring. The male
is a small apterous
louse found in the galls, very sedentary in habits. Earwigs causing Annoyance. Earwigs causing Annoyance Indoors. Mr. F. W. Carter,
of the Board
of Agriculture,
^^Tites
that
his
house
at Boxmoor
is infested with Earwigs (31.vii.02), which come
into the house each night
in
large quantities
; they appeared
to be
living in tlie crevices between the window-frames and the brickwork,
and also they seemed to be in the soil. What
I am anxious
to
know,
A\Tites Mr. Carter,
is what they feed
upon, their
habits, etc., and
also what,
if any, chemicals could
safely be
employed
to
eradicate
them
from
the
window
frames. I
have
tried
syringing with paraffin and water, but no use. I have also tried spreading
unslacked lime under the window
sills
to prevent them creeping into the
house, but
of no use. Can you suggest any means
of eradicating them,
etc. The following reply was sent
: " Judging from the description you give of the Earwig nuisance, I
should say the species is the large Earwig {Forficula auricular ia), which First Report on Economic Zoology. I20 seldom flies. A smallei' species, Lohia iiiinor, uses its wings much more
freely. The food of earwigs is very Aaried, they are both carnivorous
and herbivorous
; hops,
fruit,
snails, slugs, flowers, leaves, etc., fonii
their bill oi fire as a rule. " Tliey are mostly nocturnal in liabits, hiding away duiing the day
in
cre\ices in
walls, woodwork,
etc., under
the baik of
trees, under
clods of earth, and any shelter upon the ground. They lay their eggs
in a hole
in
the ground, about twenty to
thirty yellowish ova being
placed together. Some
authorities say the female looks
after
these
eggs
and
tlie young
for some
time. They take from two
to
four
weeks to incubate. The young are
at
first very pallid and wingless,
but
after
several moults they reach the mature winged form. The
winter is passed in the adult stage, the insects hibernating under the
bark
of trees, beneath rubbish, etc. Tliey become
noticeable
in
the
latter part
of the summer, especially in August, l)ut may
l>e found
much sooner. The small Earwig {Lithia miiwr) flies in the day-time
as well as at night. " With regard to their destruction, 'trapping' is the most successful
method. Place some baskets filled with straw or dry moss under the
windows and some flower pots
filled with moss on the window sills. Earwigs causing Annoyance Indoors. These should be examined in the day-time and
the
insects collected
and destroyed. I
do
not think you could employ any chemical, as
they get
into
so many places of a day-time
; but by
' trapping
' you
will get rid
of the nuisance, especially if you put
a plum
or other
fruit in each pot." {Gbjci'phcKjns domesticus and G. spinipcs). {Gbjci'phcKjns domesticus and G. spinipcs). Several
instances
of
acarine
pests
have
been
i-eported. Mr. White WTites fr(jm liirmingham,
" This mite
is a veritalfle plague in
my house."
It proved
to
l)e
the
Glyr.lphag as domesticus, De Geer
(the G. cursor of Ger\'ais). Another correspondent wrote from Walthamstow as follows
: Another correspondent wrote from Walthamstow as follows
:
" I have upholstered a suite for my firm which has been sent into
the country, and since it has been in the customer's possession it has
developed a small insect as sample sent on enclosed piece of banding
. . . my firm seem to think
I have been using dirty material, but
it. is nothing of the kind." The
mites
sent
were
identified
by
Albert
Michael,
Esq.,
as Furniture Pests. 121 Glyciphagiis Hpinipa^ of Kocli, and G. dumcstln'x, De Geer. G. apinipefi
is an abundant and
widely distributed mite, and
feeds
chiefly on
dried animal and vegetable matter. It
is found abundantly in stra\s
and hay, also in flour, meal, cantharides, horsehau; etc. G. doincsticus
is
also an abundant acarus
in
houses, sheds, stables, etc., and
feeds
on hay, straw, bran, on
dried
fruits, dead
insects,
cork, tobacco, and
unclean horsehair. It is frequently found in furniture. Oudemans
found
it " literally covering the furniture
of the whole house," and
states
that they fed on
the
animal
fat which adhered
to the
not
thoroughly cleaned
horsehair with which the
furniture was
stuffed. ' B
ld
itk ' B A
' B
Fig. 15. household mitks. A. O'li/ciphagiis iloinest ictis (De Geei). B. G. S2niiijies (Koch). (After Albeit Michael.) A B Fig. 15. household mitks. A. O'li/ciphagiis iloinest ictis (De Geei). B. G. S2niiijies (Koch). (After Albeit Michael.) B. G. S2niiijies (Koch). It
is thus
likely they often originate from
the
stuffing
used, l)ut
it
is not possible to say. It
is thus
likely they often originate from
the
stuffing
used, l)ut
it
is not possible to say. Nine
species
of
this genus
of
mites
occcur
in
Great
Britain. Three
species (G. dispar, Michael
;
G. crameri, Michael
; and G. pla-
tygaster,
INIichael)
live
in moles'
nests
; one {G. semrus, Haller), in
squiiTel
nests
;
the
others
{G. palmifer,
Eol)t. Fum. ;
G. canestrini,
Armanelli
; and G. pUDiiujcr, Koch), commonly in
staltle fodder and
in dust and sawdust. LiFE-HISTOKY. The
wind may also spread these minute creatures in other stages as well. The " feather-bristle " mites, or Glyeiphagi, may often
be noticed
in
houses
suffering from
dry
rot. They do not
seem
to do any
material damage in a house except
to stored
goods, unless
it
is by
carrying the
spores
of dry rot fungus
about. Mr. White
stated
in
one
of
his
communications,
" the
white
insects
give
considerable
trouble on my furniture
; perfectly harmless, but unpleasant."
They
may also be noticed in abundance in furniture attacked by the furni-
ture pest—the Death Watch (Anolium tessaUatum)—living amongst
the dust and debris these pests produce. The " feather-bristle " mites, or Glyeiphagi, may often
be noticed
in
houses
suffering from
dry
rot. They do not
seem
to do any
material damage in a house except
to stored
goods, unless
it
is by
carrying the
spores
of dry rot fungus
about. Mr. White
stated
in
one
of
his
communications,
" the
white
insects
give
considerable
trouble on my furniture
; perfectly harmless, but unpleasant."
They
may also be noticed in abundance in furniture attacked by the furni-
ture pest—the Death Watch (Anolium tessaUatum)—living amongst
the dust and debris these pests produce. Your
letter has not been answered
because
I was waiting
the
result
of some experiments
Prof.
Hall
Avas
making
for me
re
bisulphide
of
carbon.
He finds
it
will not hurt gold
picture frames, etc., unless there
are uupurities in the gold.
Nor will
it hurt furniture, foods
or draperies LiFE-HISTOKY. These minute
acari
deposit
their
eggs amongst
the substances
upon which they feed. The
eggs
are
comparatively laige, oval and
smooth-shelled,
of
a
dull
grey
or white,
the outer covering
])eing First Report on Eco)W7nic Zoology. 122 more or less
soft, not a hard, rigid
shell. The egg gives rise to the
so-called larval
stage, which resemhles the
adult, except that
it has
six instead
of eight
legs, and they are usually colourless and semi-
transparent. This
larval stage does not
last
long,
a
single ecdysis
bringing it to the third or nymph condition. The nymph resembles the adult when n earing maturity, but when
young it has the appearance
of the
larva. This
is the period when
the mite grows, and
it assumes
its
fourth
pair
of
legs. The nymph
casts
its skin
twice. Another
curious
stage
exists
in these
acari,
namely, the hypopial
stage, in which the mite assumes
a
different
appearance—a stage in which
it can more easily be distributed from
place to place by becoming attached to flying insects, etc., but in the
two
furniture pests
this
stage
is
rudimentary
; in
G. domcsticus
it
never emerges from the young nymphal
skin, and
in G. »pinipcs
it
seldom does
so. This
" skin-like "
case
protects the mite and
so
enables it to withstand heat, moisture, etc., and in
this stage
it may
very easily be
distributed from
place
to place by
the wind. The
wind may also spread these minute creatures in other stages as well. The nymph resembles the adult when n earing maturity, but when
young it has the appearance
of the
larva. This
is the period when
the mite grows, and
it assumes
its
fourth
pair
of
legs. The nymph
casts
its skin
twice. Another
curious
stage
exists
in these
acari,
namely, the hypopial
stage, in which the mite assumes
a
different
appearance—a stage in which
it can more easily be distributed from
place to place by becoming attached to flying insects, etc., but in the
two
furniture pests
this
stage
is
rudimentary
; in
G. domcsticus
it
never emerges from the young nymphal
skin, and
in G. »pinipcs
it
seldom does
so. This
" skin-like "
case
protects the mite and
so
enables it to withstand heat, moisture, etc., and in
this stage
it may
very easily be
distributed from
place
to place by
the wind. for 1,000 cubic feet of space. for 1,000 cubic feet of space. Proceed as follows
: Add the 4 ozs. of acid to the 7 ozs. water in a
deep saucer or jam-pot
; then
roll up the small lumps of cyanide in
blotting-paper and drop
into the acid and water. Leave for a couple
of hours
; then freely ventilate the room
; do not enter it for an hour
after ventilation, as, of course, you must not breathe the fumes, as they
are deadly, and
so
is the cyanide. It would
be
safest to bury the
residue, but
it
is innocuous. You
can
easily manage windows
for
ventilation, so as not to have
to enter the room to do so. I
should
put the saucer just inside the
door,
so the arm can reach
it,
and
drop the cyanide in blotting-paper into the
saucer, shutting the door
immediately. Of course, do not
let people stand about outside the
door, as some fumes may come through
crevices, etc. Get the room
as air-tight as possible. The proportions I give have been found sufficient in greenhouses
to kill Eed Spider, Woodlice, Slugs, Aphis, and Caterpillars. The employment of this gas for Bud Mite in Currants {Eriophyes
rihis) has not proved it to be successful. Sulphur in some form alone
seems to affect
acari. It
is thus interesting to learn that fumigation
with hydrocyanic acid gas did not affect this household pest, but Mr. White tells me he cleared it out by sulphur fumigation, Tkeatment. Washing well
all
likely corners where they may shelter with a
strong
solution
of
" Chinosol "
was
recommended. Failing
this,
fumigation with hydrocyanic acid gas
or disulphide
of carbon might
be tried. These, of course, are poisonous, also
the fumes, and
bisul-
phide
of
carbon
is
also inflammable,
so must
be used with
care
{vide p. 126). A further
letter regarding fumigation
for
this pest was received
from Mr. Howard White, to which the following answer was sent
: Your
letter has not been answered
because
I was waiting
the
result
of some experiments
Prof. Hall
Avas
making
for me
re
bisulphide
of
carbon. He finds
it
will not hurt gold
picture frames, etc., unless there
are uupurities in the gold. Nor will
it hurt furniture, foods
or draperies Furnitttre Pests. 123 if well aired afterwards
; but as it is highly explosive he advises the use of
hydrocyanic acid gas as used for destroying insects under glass, etc. The cyanide treatment
is deadly
to
all
insect
life and does not harm
food or anything
; but care has to be taken that the poison is not eaten or
the fumes inhaled by any person. You could only use the Chinosol for floors and crevices. Directions for using Hydrocyanic Acid Gas Indooi's. 7 ozs. of water, 7 ozs. of water, Directions for using Hydrocyanic Acid Gas Indooi's. The following
are instructions sent to Mr. White with regard
to
the gas treatment. The proportions for hydrocyanic acid gas treatment are as follows 2 ozs. of cyanide of potass 2 ozs. of cyanide of potassium 4 ozs. of sulphuric acid. Anohium tessellatum in St. Alban's Cathedral. An insect, sent by Mr. Nathaniel Hicks, in oak from the
roof
of
St. Alban's Cathedral, proved to be one
of the common Wood-boring
Beetles—a
serious
furniture
pest—known
as
the
" Death Watch
{Anohium tessellatum). First Report on Economic Zoology. 124 Judging from
tlie destructive
habits of
this pest aud
the
great
difficulty in ridding furniture
of
it when once
it gets well into the
wood, it is extremely dangerous to use such wood unless it
is treated
to destroy all the pests in
it first. For
treatment
:
corrosive
sublimate
alone
seems
of any
avail. Paraffin has been found practically useless. The Indian Meal Moth (Plodia
interjnmctella, Huebn.)
attacking Almonds. Almonds
sent
to
the Museum from
stores
in
the Docks, proved
to be attacked by one
of
the Meal
Moths
(Plodia
intcrpnnctella,
Huelm.), popularly called the Indian Meal Moth. The Indian Meal Moth caterpillar often spins a mass of
silk such
as sent with the attacked almonds. It is recorded as attacking beans,
peas, i)eanuts, walnuts, dried
fruits, almonds, and various other dried
products, including cinnamon-bark, dried dandelion roots, etc. The moth is about three-fourths
of an inch
across the expanded
wings
; some specimens, however, only reach half an inch. The outer
two-thirds of the front wings are
dull
reddish-1 )rown
; the basal part
and all the hind wings dull grey. The moth
deposits her eggs oxev
the articles destined for larval food, and also on boards, on walls, and
on floors ;
these white oxa are laid both singly and in groups of from
three to twelve. In a few days, variously estimated at from three to
seven, they hatch. The larva varies from dull white
to pale reddish
or dull yelhjw, with brown head, and
is more or
less
hairy. When
mature
it
reaches
half an inch
in
length, and then
spins a
loose
cocoon in which
it changes
to
a pale
l)rown pupa. The M'hole life-
cycle takes lour or fi\'e weeks
;
S(j that a number of broods may appeal-
under favourable conditions. Fumigating with liisnlphide
of carbon, or hydrocyanic
acid
gas,
is the only treatment. Notes and
instructions on fumigation with bisulphide
of carbon
and hydrocyanic acid gas are appended; neither harm food
if freely
ventilated afterwards {vide p. 126). Cigar Beetle and Larder Beetle. 125 The Cigar Beetle. {Lasiodcnna
tcsfacca,
l)uf.) Dr. Cliristy,
of
St. James Place, forwarded some Indian
cigars, a
case of which were l)eing
seriously damaged hy a small
l)eetle. The
pest
is kuown
as the
Cigar
JJeetie [Ludodemui
tcdacca,
Duf.), and
is closely related to the American, West Indian, and almost cosmopo-
litan Cigarette
Beetle {Ladoderma
>icrrworne). It
is
recorded from
India as attacking cheroots, rice, saffron, the
leaf coverings of opinm
balls, etc. They
have
l)een
noticed
largely
in Burmah
cheroots,
and
are
frequently found in ^lanilla cigars and cheroots. It is almost cosmo-
politan,
and
attacks
most
dry
vegetable
substances. I
do
not
remember having seen
it in Havana cigars. The egg stage lasts from
eight to sixteen days. Tlie grub
stage normally takes six weeks, but
under
unfavtniraljle conditions
it may last
for a year or more. The
pupal stage lasts about eight days. The beetles may live
for two months. The development
of the
larva'
is hastened and retarded by heat and
cold. Both
larvic and
beetles do the damage
;
a single tunnel into the cigar, as a rule, stops
it drawing, so that the damage done by a comparatively small number
of beetles and tlieir
larvio in a case may often be
consideral)le. This
pest, when
a
case
is opened and found
to
be
infested, may
easily
be checked by bisulphide fumigation. Instructions for the Fumigation of Stores with Bisulphide of Carbon. 1. Obtain the best bisulphide of carbon. 2. Eemember that
it
is both poisonous and highly inflammable
;
no light of any kind should go near
it, nor should it
l)e used where
electric wkes run, 3. Dried goods are best placed in a
large
air-tight
bin, and
then
the bisulphide placed in
saucers on
the
substance
to be fumigated. The bin should be closed and kept shut
for four
or
five hours
;
the
treated commodities should then be freely ventilated. 4. The quantity to use is 1 lb. to every 1000 cubic feet of space. If you are much troubled with
insects
in
stores it would be well
worth
having
large
bins
(air-tight) made
for
the
treatment. The
vapour given
off
is heavier than
air, and hence penetrates into the
stuff below. As there may be eggs (which I do not think are affected), a second
fumigation two weeks later would be advisable. You want to pour the bisulphide
out
rapidly, so
as not to inhale
much of the fumes
; a small quantity breathed in will not
affect one,
but it is well to be very careful. Treatment
in
bins
is
far
safer and
better than
fumigating
the
whole room, which may be well cleaned out by scrubbing with
hot. soft-soap and water. The Larder Beetle. (DermcstiS
lardarins, Linn.) The
Larder
Beetle previously mentioned
(p. 45) was
sent with
various enquiries ])y a correspondent from Wantage. This
beetle
is
also known
as
the Bacon
]>eetle. As
far
as
personal observations go it lays its
eggs
first in INIay and on through
the year in successive broods, under favourable conditions. No observations have been made on the length of
egg-life. The
larvie that
I have kept under observation took nearly five weeks
to
reach maturity, but I believe they may do so in four weeks. Hams and bacon
affected
by
this
pest
should
have
the
parts
invaded by the
insects
cut away and washed with a strong solution
of salicylate of soda. Store rooms
in which
this
pest
lias occurred
should be weU swept
out and
either fumigated with bisulphide
of
carbon or hydrocyanic acid gas. First Report on Economic Zoology. 126 Instructions for the Fumigation of Stores with Bisulphide of Carbon. Hydrocyanic Acid Gas. The fumes of this gas are also deadly poisonous to all animal life,,
with the exception, it seems, of Mites or Acari. It is safer in one way
to use than the former, owing to its not being inflammable. It
is formed by
the
mixture
of (1)
cyanide
of potassium;
(2)
sulphuric acid
; and (3) water. The fumes do not harm substances for food
; but in applying
this,
remedy you must
be
careful the men
do
not
breathe
the
fumes. Eooms can be fumigated, and the
stores in bulk, in
bins, as
before. The following rules should be remembered
: (1) Cyanide
of potassium
{a) and the fumes when mixed with
sulphuric acid (l) are very poisonous. Therefore do not breathe the
latter. (2) Use the following proportions
: (2) Use the following proportions
: 2 ozs. of cyanide, 4 ozs. of sulphuric acid, 7 ozs. of water, for every 1,000 cubic feet to be fumigated. for every 1,000 cubic feet to be fumigated. General Subjects. 127 (3) Work
as
follows
: Add
the 4
ozs. of acid
to
the 7
ozs. of
water in a saucer (never add the water
to
the acid)
; then put the
2 ozs. of cyanide in small lumps wrapped up in blotting-paper
into
the saucer and close the bin up quickly
;
the fumes do not come off
for a few seconds, so there need be no danger of inhaling them. If
fumigating a room, put the saucer close to a door and drop the cyanide
into it and shut the door rapidly
; manage
so that you need only put
your arm into the room. Leave
the
stores
in the fumes
for two hours
at
least, and then
open the windows from outside
; do not go into the room for at
least
an hour after the doors and windows have been thrown open. Night-
time is the best time to fumigate
;
a man
should keep guard to stop
people going near when
either method
is employed, if many people
are al^out and used to entering the room. If you can put the stores in
bins, I should use bisulphide
;
if to
fumigate a room or store, the hydrocyanic gas. With reasonable care
there is no danger in the use of either. Fi/st Report on Economic Zoology. Fi/st Report on Economic Zoology. 128 Suh-Oroup A.
Animals Injurioxis to Domesticated Animals. Filariasis in Lambs. (W. H. Hammond, Esq., Canterbury.)
A Parasite in Fowls' Eggs. (Dr. Humphrys, Marycliiu'cli, Torquay.) Filariasis in Lambs. (W. H. Hammond, Esq., Canterbury.) A Parasite in Fowls' Eggs. (Dr. Humphrys, Marycliiu'cli, Torquay.) A Parasite in Fowls' Eggs. (Dr. Humphrys, Marycliiu'cli, Torquay.) Gkoup
F. The Death Watch (Anohium domestkuni), which were
reported
as appearing
in
large numbers on the walls of a room
that had been shut up for a year
full
of boxes, at Eastbourne. (Miss E. Branscouibe.) The Death Watch (Anohium domestkuni), which were
reported
as appearing
in
large numbers on the walls of a room
that had been shut up for a year
full
of boxes, at Eastbourne. (Miss E. Branscouibe.) The Clothes Motli (Tineola
hisrllielht), also
reported V)y the same observer, with a
note that " They do not fly about as ordinary moths, but sit on the walls and
ceilings with folded wings, waiting
for
nie to
kill them. Sometimes I kill
ten in a room, tlien find uonn for a day or two
; then eight or ten make their
appearance
in
a room which
is shut up and no window
opened. I am
thinking
of
shutting up
the house,
so
it
is
important
for me
to know
what to d<>."
(Vide Report, p. 43.) Method
of
destroying
insects
(sp. (?) )
in
Acacia
wood. (T. Christy &
Co.,
London.)
Letter ct)ntaining a note
that " they always submit
drugs with
weevils and insects
in
theru
to
a
jirocess
of
baking."
AVe have
therefore
made arrangements for tliem
to
receive the Avhole
of om-
parcel
(of acacia
wood) to treat
it in the ordinary way. Green Matter in Lewes Public Bath. To an enquiry received by the
Director from the Lewes Town
Council regarding the inconvenience
caused by quantities of green
matter appearing in the Public Baths, the following report was sent
: The
green
matter sent from
the
public
bath
at
Lewes
is
mainly
composed
of
algte—the
majority
are
Desmids
and
Diatoms—but
the
slime masses are formed by a
Leiieoiiostoc. There
are no traces of any of
these in the sample of water sent taken direct from the pump. The bottom and
sides
of
the
bath are probably covered
witli
these
organisms, which increase with great rapidity
; the slime masses formed by
the Leuconostoc forming on the
floor and
sides
of the bath, and when
disturbed by swimmers float to the
surface. The green colour
is due to
certain
of
the
" blue
green "
algae
contained
with
the
Desmids
and
Diatoms
in the slime. A few protozoa and dipterous
larvae and seeds of
elm, etc., were also present, but the cause
of the green floating masses is
undoubtedly the slime-forming alg» referred to. The
remedy found
successful
in
cases
of
a
similar
nature
is
the
employment of sulphate
of
lime. The bath should be well cleansed and
washed out a few times with a strong solution
of the above, the sides as
well as the floor. GKOur B. Sericulture, list of works on. (H. A. Kelly, Casale Litta, Lombardy.) Suh-Oroup B.
(Section HI.)
Forestry. Suh-Oroup B. (Section HI.)
Forestry. Cecidomyia
salicis, on Willows near Canterbury. (Mr. W. H. Hammond.)
Gi>at
Moth attacking elm. (Brondesbury.) Sulj-Groiip B.
(Section
I.)
Agriculture: Fruit. The Wood Leopard (Zcuzcra wsculi) attacking Apple Trees at Hailsham. The Pith Moth (Lnnrria atra) attacking Api)le Shoots at Hailsham. (ilr. Bear)
;
at
Swanley. (Mr. Cecil Hooper.) The Bud Moth {Hedya
ocellana) attacking Apple and Cherry
at
Hailsham and
Swanley. Winter Moth (C'heimatohia hrumata) from Swanley. (Mr. Cecil Hooper.) Red Plum Maggot (Opadia fnnehrana) in fruit in Kent. (Mr. W. H. Hannnond.) Apple
Sawfly
(Hoj)locampa
testudinea)
at G nestling,
Sussex. (Rev. E. N. Bloomfield.) Pear Midge
(Biplosis
piyrivora)
attacking
Pears
at
Guestling. (Rev. E. N. Bloomfield)
;
at Swanley (Wilkinson)
;
at Ross (Getting). General. 1 29 Scale
Insects on Currants (P. rihesix, and
L. rihis). (G,
B. King,
Lawrence)
Massachusetts, U.S.A.)
Regarding
Pulvinaria, ribesix
Professor
King
writes
as
follows:—"The
Pulvinaria is what
1
believe
to be
P. vitis, L. ;
in
fact, it
is
identical
with what
I
have
written of (MS.)
a
species common on grapes
iu
Germany." Scale
Insects on Currants (P. rihesix, and
L. rihis). (G,
B. King,
Lawrence)
Massachusetts, U.S.A.)
Regarding
Pulvinaria, ribesix
Professor
King
writes
as
follows:—"The
Pulvinaria is what
1
believe
to be
P. vitis, L. ;
in
fact, it
is
identical
with what
I
have
written of (MS.)
a
species common on grapes
iu
Germany." Scale
Insects on Currants (P. rihesix, and
L. rihis). (G,
B. King,
Lawrence)
Massachusetts, U.S.A.) Regarding
Pulvinaria, ribesix
Professor
King
writes
as
follows:—"The
Pulvinaria is what
1
believe
to be
P. vitis, L. ;
in
fact, it
is
identical
with what
I
have
written of (MS.)
a
species common on grapes
iu
Germany." Information
re Currant
Scale and
caustic alkali wasli, advising use of same
to b
continued. Good
results
reported by correspondent, J. Riley,
Esq., Putley
Court, Ledbury. Winter Moth and Grease Banding. Information
seht
to
Mr. R. Amos,
Perr\-
Court, Wye, as to date, etc., for keeping bands on the trees. Sub- Group B.
(Section II.)
Horticulture. Acari
attacking
Roots
of
Flowers
(Tetranychus). (James
Nimms,
17,
Great
Tower St.) Leaf-Cutting Bee (Megachile willoughbleUa) tunnelling Apple wood. (Sir Joseph
Hooker, per the Director, and Mr. Towns-Smith, Yalding, Kent.) Thysanoptera
:
Correspondence
regarding
Haliday's
types. (Mr. Froggatt,
Gov. Ent., N. S. Wales.) Leaf Miners (Phytomyza, sp. (?)) in Melons. (Mr. Staines, Riugwould, Dover.) To the Editor of " The Journal of Tropical Medicine." To the Editor of " The Journal of Tropical Medicine." Dear
Sir,—I
beg
to
forward
you
an
insect
which
I
shall be
glad
if you wdl kindly get identified. It inflicts a very nasty sting, which
is done by a huge proboscis capable of being folded up beneath the maxilla
and neck. Acute pain and inflammation follow in a few minutes. In one
case the whole leg became swollen. Yours, &c., Yours, &c., LiM Boon Keno, M.B., C.M.Edin. LiM Boon Keno, M.B., C.M.Edin. Singapore. General. List of books, e'c, useful
for the Herefordshire Fruitgrowers' Association. (S. E. Agri. College.) First Report on Economic Zoology. 130 (B) EXTRA-BRITISH. Animals which concern Man as causing bodily
injury, some-
times
death,
to
him,
and
in
other
cases
disease,
often
of a deadly character. A Poisonous Land Bug from Singapore. A large land bug received by Dr. Cantlie from Singapore proved
to
be
one
of
the
Hemiptera
heteroptera, known
as
Conorliinus
ritbrofasciatus, De
Geer. It
is
neotropical
and
oriental
in
dis-
tribution. Some of the
foreign bugs
are very poisonous. The note sent to
Dr. Cantlie is of considerable interest. It is as follows
: (From "The Journal op Tropical Medicine," Novevihcr
1, 1901). (From "The Journal op Tropical Medicine," Novevihcr
1, 1901). IDENTIFICATION OF THE INSECT REFERRED TO BY
DR. LIM BOON KENG. To the Editor o/"The Journal of Tropical Medicine." Dear
Sir,—The
insect you send from
Singapore
is one
of
the
Hemiptera-heteroptera known
as Conorhinus riihrofasciatus, De
Geer. It
is neotropical and oriental in distribution. Yours, etc.,
Fred. V. Theobald. Fred. V. Theobald. British Museum (Natural History). British Museum (Natural History). Screw Worms
in Human Beings, 131 A full account
of Hemiptera-heteroptera obnoxious
to man
lias
recently been
compiled
by Eaphael
Blanchard
entitled,
" Sur
la
Piqiire de quelques Hemipteres."
Archives de Parasitologie, V. No. 1,
p. 139 (1902). SUB-GEOUP
A.
ANIMALS WHICH CONCERN MAN BY
CAUSING
BODILY
INJURY
OR
DISEASE
TO
HIS
STOCK OF DOMESTICATED ANIMALS. SUB-GEOUP
A. ANIMALS WHICH CONCERN MAN BY
CAUSING
BODILY
INJURY
OR
DISEASE
TO
HIS
STOCK OF DOMESTICATED ANIMALS. GROUP
E. GROUP
E. Animals which
concern Man
as
causing
bodily
injury
or
disease,
both
possibly
of a deadly character,
to
(A)
his
stock of Domesticated Animals, or
(B)
to
his Vegetable
Plantations, or
(C)
to Wild Animals in the
preservation
of which he is interested, or (D) Plants in the preservation
of which he is interested. GROUP
E. Animals which
concern Man
as
causing
bodily
injury
or
disease,
both
possibly
of a deadly character,
to
(A)
his
stock of Domesticated Animals, or
(B)
to
his Vegetable
Plantations, or
(C)
to Wild Animals in the
preservation
of which he is interested, or (D) Plants in the preservation
of which he is interested. Screw Worms
in Human
Beings. A note regarding
the Screw Worm {Compsomyia macdlaria)
in
human beings was sent by Dr. St. George Gray. Screw Worms were
taken from the mouth and nose of a female patient dying of phthisis
in the Victoria Hospital. " I am not
at
all
sure that they are the
larvae of Compsomyia
macellaria, which
is
very common
in
some
parts of this island and which attacks cattle, for I have never seen a
single specimen
of the adult
fly in the vicinity of the hospital, but
there are innumerable
flies
of other
species
about. I am trying to
breed out a few of these in a jar of earth, so that I may be absolutely
certain of the fly. I may mention one
fact about them which may
be
of interest. Out
of four
patients who were attacked by Screw
Worms two occupied the same bed, one after the other, and a third
the next bed
to
it. The other case was in a more remote part of the
hospital." About a month after, specimens of the
fly and its puparia arrived
with
the
following interesting notes by Dr. St. George Gray
:
" I
notice that those bred from the
larvse have a more decided bluish
colour than those caught
in
the open. This may
be
due
to
the
fact that the former had never
fed. During
life the eyes
are
of a
brick-red
colour,
but
this
changes
after
death."
The
larva? of
C. macellaria have
frequently been known to attack human bemgs,
entering the external orifices. K
2 First Report on Economic Zoology. 132 Bulletin
of the Agricultural Station
of Louisiana, No.
2, second
series.
"The Texas Screw Worm," by Prof. H. A. Morgan, 1890.
All animals seem to be attacked by
it. Section
I. Animals Injurious to Agriculture. Pony
Flies. {Lyperosia, sp. ?) Mr. E. E. Green forwarded some small
flies
that were
causing
annoyance
in the pony-breeding
establishments
in
Ceylon,
They
were examined by Mr. Austen and found
to belong
to
the
family
Muscidae and
to the genus Lyperosia, sp. (?). The species is probably
new. The Screw Worm in Cattle in St.
Lucia. Two diptera sent by the Director of the Imperial Department of
Agiiculture of the West Indies that are injurious to cattle in St. Lucia
proved
to
be the well known
" Screw Worm " Ely. The
scientific
name
of the
fly
is
Compsomyia
macellaria,
of Fabriciiis. There
is
any amount
of
literature
on
this
pest,
its
life-history being well
known. The fly is common from the Argentine to Canada. It especially
attacks the natural openings
of animals, notably the
" sheaths "
of
horses and the navel of newly -born animals
; but the fly will lay its
egg upon any abraded surface of the
skin. Wliere
ticks, etc., have
been killed on an animal
is
a favourite place
for the fly to deposit
her
eggs,
the
fly being
attracted
by the
blood. Abrasions
from
contact with barbed wire form
favourite
localities. Human beings
are
also subject to
its
rarages, especially in the nose and
ear
{vide
"Psyche"
iv.,
pp. 27-30,
1883,
and
page
131). Amongst
the
many
excellent
account|
issued
by
American
stations
is
the
following
:
\ Bulletin
of the Agricultural Station
of Louisiana, No. 2, second
series. "The Texas Screw Worm," by Prof. H. A. Morgan, 1890. Bulletin
of the Agricultural Station
of Louisiana, No. 2, second
series. "The Texas Screw Worm," by Prof. H. A. Morgan, 1890. All animals seem to be attacked by
it. All animals seem to be attacked by
it. Pony Flies and Scale Insect. 133 Scale Insect
{Mytilaspis
cifricola, Packard) on Orange
Trees in Monte Video. Dr. E. S. Miller,
E.N., sent
from
Monte Video a scale
insect
affecting
the
orange
trees
there and
asking
for information
as
to-
destroying
it. This
scale proved to be Mytilaspis citricola, Packard. It occurs in the United
States, West
Indies, China, Brazil, Southern
Europe, Ceylon, Eiji, etc. It has been recently introduced into South
Africa. Fruit from Southern Europe, Canary and Madeira is usually
infested. Its food plants are all citrus fruits and probably all llosaceaj. In
Jamaica, Cockerell records it on the Murraya. Its original home was
probably the West Indies or South America. It occurs on leaf, fruity
stems and twigs. This scale is about ^th
of an inch
long, and
is about three times
as long as
it
is wide, and
like the Apple Mussel scale in outline, the
anterior end being narrow and the
posterior broad and rounded, the
whole scale somewhat curved. The
colour
is
variable, some
are
dull
purplish,
others
almost
brown. Beneath the scale
is white
;
this lower white portion coming
away with the scale retains the insect or eggs within. The male scale is almost straight and ^th of an inch long. The
eggs, which vary from
twenty-five
to
seventy under each
scale, are white. All the specimens examined from Monte Video had
eggs within them. First Report on Economic Zoology. 134 Three or four broods may occur iu tropical climates. It is known
under a variety of popular names, such
as
the Purple Scale and the
Orange Mussel Scale. Treatment of Affected Trees. There are two Avays of treating scale-insect attack
: There are two Avays of treating scale-insect attack
:
I. Spraying, with either paraffin emulsion or resin wash, I. Spraying, with either paraffin emulsion or resin wash, II. The gas treatment. 1. Spraying for scale attack. 1. Spraying for scale attack. A. Paraffin
emulsion. This
is
used
to
kill
the
larval
scale
insects when
they are
crawling
from
beneath
the
scales
and
to
corrode the scales away. The time of migration of the larv?e should
be noticed when this wash
is
used, and the wash applied when they
are crawling about, to do most good. But
it has been found more or
less
satisfactory at any time
if applied on
several occasions during
the year, best at intervals of two weeks. Paraffin emulsion
is made
as
follows
:—Mix
equal
portions
of
soft
soap
dissolved
in
boiling water and
paraffin, and
then churn
them up by means
of
a
force pump
until a creamy emulsion
is
produced. When
required
for use mix with twenty times
its bulk
of water, B. Kesin
wasli. There
are many recipes for this—the following
is a good one
:—dissolve
1
lb. of caustic soda
in 1^ gallons of soft
water, then dissolve 2
lbs. of resin and
1
lb. of tallow by moderate
heat, and
as
it is cooking
stir in gradually 1 quart
of the dissolved
caustic soda solution, and then add water until you have 22 pints of
liquid. This forms
a thick brown soap which
is
sufficient
for 44
gallons of wash
;
it being added
to the water and well stirred
; warm
water,
if possible, should be employed. This should be applied before
the blossom
bursts,
but
in bad
attacks
it may
be
used
later
as
it is
well
to
clear out the
scale, even
at the loss of
all that
year's
crop, through destroying the blossom, 2. Gas
treatment. This
is
largely employed
for
scale
in
the
United
States, the Cape
of Good Hope,
etc. The
trees have
to be
covered with a tent
of gas-proof canvas
; the hydrocyanic acid gas is
generated by putting water and sulphuric
acid
in
a
saucer
or
jar'
and then putting lumps
of 60 per
cent, grade cyanide
of potassium
into
the
mixture. The fumes may
be
allowed
to
ascend
for an
hour or more—evening or dull days
are the
best times to fumigate. The slower the gas generates
the
better
;
the greater the quantity of Scale on Pineapples and Pine Beetle. 135 water and the
larger the lumps
of cyanide used the slower the gas
comes
off. Experiments conducted by Mr. {Dias-pis
hromdicc,
Kerner.) Pineapples are frequently damaged by a
scale
insect, which now
and then causes the fruit to
rot. Specimens have been
sent by Mr. Hammond, of Kingston, Jamaica,
for
identification and information
concerning it. This scale
is the Pineapple Scale (Diaspis Iromelite,
Kerner). The
scale
is
tliin,
circular and
pure white—the females
yellow
or
orange. Like
most
Diaspids, they
burrow beneath
the
epidermis of the plants and become almost entirely hidden. It chiefly attacks the leaves, but now and then the fruit. It should be destroyed as soon as the fruit is cut. It chiefly attacks the leaves, but now and then the fruit. It should be destroyed as soon as the fruit is cut. 1. Spraying for scale attack. Cousins
in England show that the
following quantities
are required per cubic
foot
of space—3
ozs. of
cyanide, 5 ozs. of acid, 8 ozs. of water per 1000 cubic feet. Both the
cyanide and the gas generated are poisons. If gas treatment cannot be
carried
out owing to lack of material
or apparatus, then spraying should be employed. "Manual of Forestry," Vol. IV., p. 242. {Hylesinus pinipei^da, Linn.) Some pine wood sent by R. E. Haslam, Esq., from Monico, was
found to be attacked by the Pine Beetle {Hylesinus piniperda, Linn.)
It had killed an old
tree and two young
ones. This insect
chiefly
attacks diseased and damaged timber
; but
if no unhealthy trees are
about
it
will
attack
healthy
ones. As
a
rule
one
sees
this
pest
working in plantations of about thirty years standing. <> Sootch and Weymouth Pines are chiefly attacked, but cluster and
other species
of pines
are frequently recorded
as being damaged by
this
pest. This insect does harm
in
three ways
;
(i) the
beetles and
larvse
attack ])ark and
bast, the former making longitudinal
galleries with First Report on Economic Zoology. 136 one to three
air holes — the
larvic
eat out secondary galleries in the
bast which branch out at right angles to the primary gallery
;
(ii) the
beetles in August and September bore
into the pith
of young pine
shoots at a distance of from one to three inches from their extremities,
eating out a tunnel up
to
the terminal bud. The holes wliere the
entrance is made are surrounded by a ring of opaque resin
;
(iii) the
beetles bore into the sap wood
of the root-stock of quite sound trees
to hibernate, and thus
trees may become
sickly that were formerly
quite sound, and so attract l^eetles during the following year. The
beetles appear
in April and May, and again
in June and
July. The
larvse hatch
in April and May, and pupate in June or
July and even August. Those that hatch in June may produce a second brood in August,
and this second brood attacks the terminal shoots and branches. The
whole life cycle lasts from sixty to eighty days. It should be remembered that the
beetles hibernate
in the adult
stage
in
the
root -stocks
and
roots
of
standing
trees,
also in old
stumps. Schlich * gives the following protective and remedial measures
: i. Timely and frequent thinnings
of woods and quick removal of
all sickly wood. ii. Clearance of felling areas by the middle of April. iii. Uprooting of stumps and broken trees or barking the same. iv. Pine woods if damaged by fire should be felled. iv. Pine woods if damaged by fire should be felled. V. Insect-eating mammals and birds should be protected. V. {Hylesinus pinipei^da, Linn.) Insect-eating mammals and birds should be protected. vi. All standing trees containing larvte and pupte should be felled
and barked and the bark burned. vi. All standing trees containing larvte and pupte should be felled
and barked and the bark burned. vii. Trap trees should be felled from February to September, so as
to supply
trees which
are not
too
dry for the
beetles
to
Ijreed
in. These should be barked at the middle of May, and others at intervals
of four to six weeks, and the bark burnt. Of these rules
the most
important
are
; (A)
the destruction of
attacked
trees
at the proper time
; and (B) using certain unliealthy
trees
as
"trap"
trees. If
there
are
no
unhealthy
trees
in
the
plantation,
certain
of them
should
be made into
" trap "
trees by
linging the worst trees. This is done by cutting strips of bark round
the trees in the early spring so as to produce an unliealthy state, and
so
attract the beetles
to lay
their eggs and
thus
keep them away
from the surrounding ones. These " trap " trees should be burnt
later,
before
the
larvse and
pupse have matured. "Manual of Forestry," Vol. IV., p. 242. Insects Injurious
to
Coffee. 137 Beetles Damaging Coffee-Berries. Two samples of coffee-berries damaged by small beetles have been
received during the past year. One was sent by a correspondent in
London with the foUowiEg information
:
" I enclose a small sample
of coffee taken from
a bag recently
in, from
Costa
Eica, and
shall
lie glad
if you can
tell me the life-history of the creatures with the
coffee, and whether they are likely to spread
to
other goods
in
the
warehouse, either coffee or cocoa or goods
of a kindred nature."
On
examination
the
sample was found
to contain
a number
of
live
beetles, and most of the berries were greatly damaged. C
A
B
Pig. 16. B, Coffee damaged by (A), Arxoctrm fasciculatus, De Geer (cf and 9)
; C, by Scolyti, sp. (?) C C Pig. 16. Pig. 16. B, Coffee damaged by (A), Arxoctrm fasciculatus, De Geer (cf and 9)
; C, by Scolyti, sp. (?) g. 6. B, Coffee damaged by (A), Arxoctrm fasciculatus, De Geer (cf and 9)
; C, by Scolyti, sp. (?) The
beetles sent
are known
as Arccocerus fascicidatus, De Geer
(Fig. 16,
a). Their
life-history
is well
known. They
are cosmo-
politan and abundant in the Old and New Worlds. The larvfe live in coffee-berries, and will attack cocoa, ginger and
other commodities, as also will the adult beetles. It
is well known in Central America, in India and
the Cape of
Good Hope, but
does
not seem abundant
in Europe. It has
also
been
found
on
Tamariscus
gallicus
and
in
and
on
packages
of
Tegenaria. The larva is short and cylindrical, with distinct legs and
about one-fourth of an inch long
;
its movements
are
slow. It has First Report on Economic Zoology. 1 38 been observed in branches of a kind of ginger from China, eating the
woody
parts, making long
galleries
deep into
the branches,
wliich
become
full
of dust
(frass). Wlien they are ready
to pupate they
make a large nest or cell near the bark, so that the beetle can escape
easily. The pupal stage
lasts from ten to fifteen days. The beetles
are very agile, jumping often nearly an inch
; they
also
fly
readily,
so that
they might soon spread themselves over a storehouse. They
also
feign death when touched. They were breeding in
the
coffee
berries
sent
as well
as having damaged them by eating them. Beetles Damaging Coffee-Berries. If
such a consignment is not destroyed,
it should be treated at once
to
kill
the
beetles. This may be done
either by
heat (if possible in
this
case) or by fumigating the mass with hydrocyanic acid gas in
closed receptacles. The
life-history
of
this
pest
has
been
fully
described
by
M. E. Lucas in
tlie Annales of the Entomological Society of France
(tom. 1, 4th se., p. 399, 1861). The
other sample of coffee
berries (Fig. 16, c) was
received
at
Kew from Uganda and sent on to the British Museum. The follow-
ing note was sent back to Kew regarding the cause of damage
: " Some coffee berries from Uganda have been handed to me by Sir
George
Hampson. They
have
been
damaged
by
Scolytidoi. Mr. Waterhouse
says
it is most probably a new species. As only a few
fragments of the
beetle could be found,
it
is not
possible to refer to
it
in
detail. Could you
obtain
fresh specimens
of
the
beetle and
have
them
sent
here ? I can
find no
record
of any
similar pest
attacking the berries. " A Ehyncota
StacJiia geometrica, Motsch (MS,)— attacks young
coffee berries in Ceylon and does some harm." These berries were eaten into, many hollowed right out, the outer
shell often perforated in two or three places. Rubber. Some Weevils
sent
by
the
Curator
of
Selangor JMuseum
that
were reported to him as
defoliating Para Rubber {Hevca
hrasiliensis)
proved to be the Hypomeces squamosus of Fabricius. Insect Pests of the West Indies. 139 Insect Pests of the West Indies. Cotton Stainer (Dysdercus audreae), attacks bolls,
-'amaica. C. Coffee. 1. Scale {Aspidiotus articidafus). Jamaica. Leaves. D. Allspice. 1. Pimento Borer {Cyrtomerus pilicornis'), bores into twigs. Jamaica. E. Banana. 1. Aspidiotus articulatus, on leaf.'\
2. Ceroplastes floridensis. 3. Aspidiotus personatus.)
4. Aspidiotus Jicus. )
Jamaica. rare. 2. Ceroplastes floridensis. 3. Aspidiotus personatus.)
4. Aspidiotus Jicus. )
Jamaica. rare. F. Congo Pea Pests. Vide No. 54, Inst. Jamaica Mus. Notes. Insect Pests of the West Indies. In answer
to Mr. Hammond, the following
list
of West Indian
insect
pests has
been
forwarded. The Orange
Tests
are
not given
in
this
list. Probably the Museum
of the Jamaican
Institute has
records, as tliey have publislied catalogues under the title of Museum
Notes. This
list
is
further
augmented
by
the
identification
of
specimens sent over by the
Imperial Department
of Agriculture
of
the West Indies. The names of these
species and their food plants
are given in the following pages. A. Insects injurious to Sugar Cane. 1. Large Sugar Cane
Borer
{Diatroea
saccharalis). (" Ins. Life," Vol. iv,
pp. 95, 103.)
2. Sugar X^leborus or Pin Borer {XyJehorus per/orans). (" Ins. Life," VtiL
V. p. 31.)
3
Sugar
Cane
Pin
Borer
{Xi/hborus
putescens)
I'arbados
Trinidad A. Insects injurious to Sugar Cane. 1. Large Sugar Cane
Borer
{Diatroea
saccharalis). (" Ins. Life," Vol. iv,
pp. 95, 103.)
2. Sugar X^leborus or Pin Borer {XyJehorus per/orans). (" Ins. Life," VtiL
V. p. 31.)
3. Sugar
Cane
Pin
Borer
{Xi/hborus
putescens). I'arbados,
Trinidad,
St. Vincent. 4. Boring Weevil (Sphenojthorus). A. Insects injurious to Sugar Cane. 1. Large Sugar Cane
Borer
{Diatroea
saccharalis). (" Ins. Life," Vol. iv,
pp. 95, 103.) 2. Sugar X^leborus or Pin Borer {XyJehorus per/orans). (" Ins. Life," VtiL
V. p. 31.) 3. Sugar
Cane
Pin
Borer
{Xi/hborus
putescens). I'arbados,
Trinidad,
St. Vincent. 3. Sugar
Cane
Pin
Borer
{Xi/hborus
putescens). I'arbados,
Trinidad,
St. Vincent. 4. Boring Weevil (Sphenojthorus). 4. Boring Weevil (Sphenojthorus). 5. Tropical Sugar Cane Borer (Chilo saccharalis). 5. Tropical Sugar Cane Borer (Chilo saccharalis). 6. Scale (in Jamaica) (Aspidiotus sacchari, Ckll.). 6. Scale (in Jamaica) (Aspidiotus sacchari, Ckll.). 7. Mites, vide
(
Histiostoma rostroserratas (decaying plants). Bull. 40, Royal
j Immature Oamasids (predacious). Kew Gardens,
j
Damacics
or
Mofaspis,
sp. Tarsonymus
hancrofti,
April, 1890. I
Miller, does damage to canes. 7. Mites, vide
(
Histiostoma rostroserratas (decaying plants). Bull. 40, Royal
j Immature Oamasids (predacious). Kew Gardens,
j
Damacics
or
Mofaspis,
sp. Tarsonymus
hancrofti,
April, 1890. I
Miller, does damage to canes. B. COTTOX. 1. Dactylohius virgatus, Ckll. = Mealy Bug on under side of loaves. Jamaica. 2. Cotton Stainer (Dysdercus audreae), attacks bolls,
-'amaica. B. COTTOX. 1. Dactylohius virgatus, Ckll. = Mealy Bug on under side of loaves. Jamaica. 2. Cotton Stainer (Dysdercus audreae), attacks bolls,
-'amaica. 1. Dactylohius virgatus, Ckll. = Mealy Bug on under side of loaves. Jamaica. 2. I. Chincona Pests. 1. Diaspis ptiargonii, Ckll. Jamaica.' 2. Lecanium decidrophthorsR, Ckll. Jamaica. F. Congo Pea Pests.
Vide No. 54, Inst. Jamaica Mus. Notes. G. Cocoa Plant. 1. Cocoa Bug {Memhracidas). Leaf-hopper found in Trinidad. 2. Leaf-cutting Ant {Alta pedeus, L.). Trinidad. 3. Longhom Beetle (Sterastoma depresstim), on young plants. 4. Palm Weevil (Rhynchophorus palmarum). G. Cocoa Plant. 1. Cocoa Bug {Memhracidas). Leaf-hopper found in Trinidad. 2. Leaf-cutting Ant {Alta pedeus, L.). Trinidad. 3. Longhom Beetle (Sterastoma depresstim), on young plants. 3. Longhom Beetle (Sterastoma depresstim), on young plants. 4. Palm Weevil (Rhynchophorus palmarum). 4. Palm Weevil (Rhynchophorus palmarum). First Report on Economic Zoology. 140 5. Diaspis hoisduvaUl. Tiinidad. 6. Fiorinia gallucida, Sig. 7. Mytitaspis buxi, Sig. = M. pandani, Cou. Abundant on leaves in Jamaica. 9.
Fiorina camellias, Cou. 9. Fiorina camellias, Cou. H.
CocoANUT Palm. H. CocoANUT Palm. 1. Autocarpis hoisduvaUi. Jamaica. 2. Aspidiotus pumicse, OsW. Jamaica. 3. Dactylopius longifilis. Jamaica. 4. Dactylopius virgatus, on leaves. Jamaica. 5. Diaspis vandalicus, Galvoz. Jamaica, abundant
; and Santiago de Cuba. 6. Rufous Scale {Aspidiotus
artictdatus), Morgan. Jamaica. (" lus. Life,"
iv. 380.)
7. Aspidiotus minutus, Ckll. 8. Aspidiotus rapax v. palmas, Ckll. 9. Fiorina camellias, Cou. 1. Autocarpis hoisduvaUi. Jamaica. 1. Autocarpis hoisduvaUi. Jamaica. 1. Autocarpis hoisduvaUi. Jamaica. 2. Aspidiotus pumicse, OsW. Jamaica. 2. Aspidiotus pumicse, OsW. Jamaica. 3. Dactylopius longifilis. Jamaica. 4. Dactylopius virgatus, on leaves. Jamaica. 5. Diaspis vandalicus, Galvoz. Jamaica, abundant
; and Santiago de Cuba. 6. Rufous Scale {Aspidiotus
artictdatus), Morgan. Jamaica. (" lus. Life,"
iv. 380.) 7. Aspidiotus minutus, Ckll. 8. Aspidiotus rapax v. palmas, Ckll. J.
Mango. J. Mango. 1. Ceroplastes fioridensis, leaves. 2. Vindonia stelli/era. 3. Aspidiotus personatus. 4. Aspddiotus articulatus. 5. Lecanium olern,
6. Lecanium mangiferae,
7. Dactylobius longifilis, Cou. 8. Aspidiotus, sp. (?), pale patches on fruit. 9. Planchonia pustulans, Ckll. Moutserrat. >
leaves. 1. Ceroplastes fioridensis, leaves. 3. Aspidiotus personatus. 4. Aspddiotus articulatus. K. POMEGKAXATE. 1. Ceroplasies floridensis, on leaves. 2. Aspidiotus pumicse, Ckll. Jamaica. Insect Pests of the
JVest Indies. Insect Pests of the
JVest Indies. 141 K.
POMEGKAXATE. L. Peach. 1. Bermuda Peach Maggot (Ceratitis, sp. (?)), attacks fruit. L. Peach. 1. Bermuda Peach Maggot (Ceratitis, sp. (?)), attacks fruit. 1. Pineapple Scale (Diasjns bromelia). N. Orange. 1. Aspidiotus citricola. Bermuda. 2. Chionaspis citri. Bermuda. 0. Animal Pests. 1. The Screw-woiTO Fly {Compsomyia macellaria). St. Lucia. NAMES OF ECONOMIC INSECTS SENT
BY THE IMPERIAL
DEPARTMENT OF AGRICULTURE OF THE WEST INDIES. Note.—(A.)
Aptera
;
(C.)
Coleoptera
;
(B.)
Blattidae
;
(L.)
Lepidoptera
;
(H.) Hymenoptera ; (He.) Hemiptera. Note.—(A.)
Aptera
;
(C.)
Coleoptera
;
(B.)
Blattidae
;
(L.)
Lepidoptera
;
(H.) Hymenoptera ; (He.) Hemiptera. Note.—(A.)
Aptera
;
(C.)
Coleoptera
;
(B.)
Blattidae
;
(L.)
Lepidoptera
;
(H.) Hymenoptera ; (He.) Hemiptera. Sugar Cane.
Myochronus armatus, B.
On leaves.
Barbados.
(C.) Sugar Cane. Myochronus armatus, B. On leaves. Barbados. (C.) Arbowboot. Calpodes ethlius. Cram. Barbados. (L.)
Megachile Jlamtarsata, Smith. Barbados, St. Vincent. (II.)
Megachile martindali, Asbmead. Barbados. (H.) Megachile Jlamtarsata, Smith. Barbados, St. Vincent. (II.) Megachile Jlamtarsata, Smith. Barbados, hile martindali, Asbmead. Barbados. (H. Andiba, sp. Cleogonus rubetra, F. Grenada and Trinidad. (C.)
Diori/merus, sp. (?) Andiba, sp. Cleogonus rubetra, F. Grenada and Trinidad. (C.)
Diori/merus, sp. (?) Banaxa. Tomarus bitaberculatus, Beard. St. Lucia. (C.) Cassava. Dilophonota ello, Linn. Montserrat. (L.) Cocoa. Gryphalus, sp. (?)
Grenada. (0.) Cowpeas. Bruchus cMnensis, Linn. Barbados. (C.) COBNWEAL. Carpophilus dimidiatus, Fabr. Barbados. (C.) Castillva elastica. Tmniotes scalaris, Fabr. Grenada. (C.) Divi-Divi (Cmsalpinia coriaria). Bruchus, sp. (to fruit),
Antigua. (C.) Ficus. Phryneta verrucosa. Baibados. (C.)
Leucoplicea surinamensis. Barbados. (B.) FiDDLE-WOOD. Pyraustra melUnaUs, Hubn. Barbados. (L.) Grasses. Eemiyia repanda, Fabr. Trinidad. (L.) Guinea Cork. Calandra oryzse, Linn. Antigua. (C.) Geandilla. LacHca pallens, Fabr. Montserrat. (C.) Indian Corn. Trogosita mauritanica. Antigua. (C.)
Aphis maidis, Fitch. Montserrat. (He.)
Spodoptera frugiperda. Barbados. (L.) First Report on Economic Zoology. 142 Mango. Platypus paralldus, Fabr. Grenada. (C.) Malpighia. Podagrica amoenissiina, Chen. MS. Antigua. (C.) Orange. Lagochiriis arauci/ormis, L. Grenada. (C.) Pigeon Pea Bush. Rhyparobia ma'hrai, F. Barbados. Pigeon Pea Bush. Rhyparobia ma'hrai, F. Barbados. Pigeon Peas
(Dried). Bnichus A-macuIatus, F. Antigua. Pigeon Peas
(Dried). Bnichus A-macuIatus, F. Antigua. Palm Seeds. Coccotrypes dadyloperda, Fabr. Trini lad. (C.) Sweet Potatoes. Cryptorhynchusi bataius, G. Waterh. To tubers. Barbados. (CI.)
Coptocycha trisiynata, Bobem, var. histripunctata, B. To leaves. Antigua. (C.)
Cheetocnemgi amazona, Baty. Eats leaves. Barbados. SOLANUM MeLONGENA. Epitrix paroula, Fabr. On leaves. Barbados. Corythaici monaclia, StS,l. On leaves. Barbados. Tamarind. Cathartus cassia?., Reich. To
1 ods. Barbados. (C.)
CryjiJialus, sp. To pods. Barbados. (C.) Tannia. Tomarus hitiiherculatus, Beard. St. Luci \. (C.) Woolly Pykol. Thermesia yemmataUs, Hubn. Barbados. (L.) Wood (Dry). 2'riholium firrugineum. Barbados. (C.)
Platypus, spp. Barbados. (C.) Wood (Dry). 2'riholium firrugineum. Barbados. (C.)
Platypus, spp. Barbados. (C.) Stored Gooub,
etc. Lepisma, sp. Starch, Gum, etc. Barbados. (A.)
Calandra oryzx, ]Ann., vsii: pallida. To macaroni. Barbadtis. (C.)
Triholiumferrugineum. To corn-flour. Barbados. (C.)
Lasioderma serricorne, F. (C.)
Ptriplaneta americana, L. (B.)
BliES. BliES. G R
U P
F. Animals
which
concern Man
as
being
destructive
to
his
worked-up
Products
of Art and
Industry,
such
as
his
various
(A)
Buildings
and
larger
Constructions
and
Habitations,
(B)
Furniture
and
Books,
Drapery
and
Clothing, (C) Food and Stores. Galleria melloncUa, Linn.
To honeycomb.
Antigua.
(L.) BENEnciAL Insects. Megilla maculata, var. De Geer. Eats various insects. Scymnus
ochroderus, Mulsaut. Feeds on
Aphis
maidis. Montserrat and
Barbados. (C.)
Exochomus nitidulus, Fab. Feeds on Coti^idse. Barbados. (C.)
Chrysopa, sp. Feeds on various insects in Barbados. (N.) 143 Teredos and Canadian Timber. A communication was
received on
23rcl November,
1901, from
the
Imperial
Institute
in regard
to
the
wood-boring
Teredos
and
Canadian
timber (Abies
canadensis). In
this
communication
the
following was
stated
:—" This Department
has
supplied
full
infor-
mation with reference to the general
characteristics of the wood and
its mechanical
properties. It appears, liowover, that other wood
so
employed
in
South Africa
is
liable
to
be
attacked by
the Teredo
worm. The Canadian wood is known not
to be readily attacked by
worms or
insects of any description
in
Canada, but the question
is,
does
this
particular
worm
occur
in
Canada ? "
The
following
replies were sent to the Institute
: (1.) The Teredo worm, one of the mollusca, attacks all manner of
wood. Canadian
fir,
judging
from
the
specimens
so
frequently
washed up on the shores of England, Norway, etc., seems very
liable
to
the
attack
of
Teredo. One
species. Teredo
megotara, Hanley,
found in England, both on fixed, floating and
drift timber (especially
in
Canadian
fir),
occurs
in
America. Stimpson
described
it
as
{T. dilatata) infesting
fixed wood and harbour buoys
at Lynn, New
England. Tryon states that
tliis species extends from Massachusetts
to South Carolina—it also occurs
in Greenland and Iceland—so lias
a very wide range, as one would expect, being ti'ansported on floating
wood, aided by the Gulf Stream. I do not know any exact
localities to give for Teredo in Canada,
but the one I mention and others occur there. I do not know any exact
localities to give for Teredo in Canada,
but the one I mention and others occur there. It
is
surprising
to
find
that Ahies canadensis
is not subject
to
boring mollusca in Canada, as drift wood of that fir appears particularly
prone to the attack of the pest. It
is
surprising
to
find
that Ahies canadensis
is not subject
to
boring mollusca in Canada, as drift wood of that fir appears particularly
prone to the attack of the pest. It
is
surprising
to
find
that Ahies canadensis
is not subject
to
boring mollusca in Canada, as drift wood of that fir appears particularly
prone to the attack of the pest. First Report on Econornic Zoology. 144 (2.) Since
I
last wrote you re Teredo and Canadian pine
I
find
the following species occur in Canada
: Tereda dorsalis (the commonest
species), Xylophaga fimhriata, and X. Teredos and Canadian Timber. hipinnata on
the West Coast,
and Teredo megotara on the East Coast. The species occurring on the United
States coast (Massachusetts)
are T. navalis, T. norvegica, T. dilatata, T.clilorotica and X. fimhriata. T. norvegica occurs in oak,
fir, and birch, and
is found
in Europe
as well. T. navalis in
fir, elm, etc. ;
also in Europe. T. megotara in
any wood
;
also European. X. hipinnata
occurs
in
Europe, West
Indies, etc., in almost any wood,
X. fimhriata has
also occurred
in
teak in Em-ope. The only
Teredo
quoted from
South
Africa
is
T. {Hyperotus)
nucivora
of Spengler, which
bores
into
floating
cocoa-nuts. This
species
Mr, Edgar Smith says he believes
to be only
tropical and
sub-tropical. Teredos which bore into wood,
floating
or
otherwise,
may be found anywhere,
and doubtless have
a
very wide
range. They may be carried very great distances by ocean currents. Other Short Eeports
sent. The
Eat
Elea {T. musculi)
;
Information
concerning
it,
and
its
connection with Plague. (Dr. Cantlie,) The
Eat
Elea {T. musculi)
;
Information
concerning
it,
and
its
connection with Plague. (Dr. Cantlie,) Tapeworms in the Bile Duct of Sheep in Transvaal (Dr, Theiler),
A new species under investigation. Tapeworms in the Bile Duct of Sheep in Transvaal (Dr, Theiler),
A new species under investigation. Hippo and
other
Flies (Tahanus
dorsovitta,
Lucilia marginalis,
and Lucilia, sp. (?)) from
Zambesi. (L, Lloyd
Prichard,
M.K.C.P.,
etc., Jersey.) Hippo and
other
Flies (Tahanus
dorsovitta,
Lucilia marginalis,
and Lucilia, sp. (?)) from
Zambesi. (L, Lloyd
Prichard,
M.K.C.P.,
etc., Jersey.) Ticks
(Ixodidce)
on
Toads
in
Para, and
their
connection
with
Drepanidium found in Toads. (Dr. Durham.) Drepanidium found in Toads. (Dr. Durham.)
Tsetse-fly in Gambia {Glossina
lonyipalpis, Wied., var, tachinoides,
Westwuod). (Dr. Dutton.)
The Swilt Tick, intermediate host of Filaria cypseli. (Dr, Dutton,)
A new species of Mallophaga,
]\Ielolonthid
Larvte
(Apogonia raicca) attacking
Coco Palms
in
Ceylon,
(E, E. Green.) Tsetse-fly in Gambia {Glossina
lonyipalpis, Wied., var, tachinoides,
Westwuod). (Dr. Dutton.) The Swilt Tick, intermediate host of Filaria cypseli. (Dr, Dutton,)
A new species of Mallophaga, ]\Ielolonthid
Larvte
(Apogonia raicca) attacking
Coco Palms
in
Ceylon,
(E, E. Green.) EEPOETS
TO
(A)-THE FOKEIGN OFFICE
AND
(B)~THE COLONIAL
OFFICE, 147 1. TSETSE-FLY AND BUFFALO CORRESrONDEXCE. FORRIGN OfFITE,
15th Jt0ie, 1901. SiE,—I am directed by tliy Marquess of Lausdowne to transmit
to
you the accompauyiug copy
of a
letter from the
Colonial
Office ou the
subject
of
the
Tsetse-fly, forwarding
a copy
of a
letter from a Mr. Val
Gieigud
to the
British South
Africa Company remarking on
the special
virulence of
this fly in districts where Buffalo are principally to be found,
and I am to request that you will favour His Lordship with your observa-
tions on the subject. I am, Sir, Your most obedient humble servant, Your most obedient humble servant, Clement Ll. Hill. Peofessob E. Ray Lankester,
British Museum (Nat. Hist.)
South Kensington, S.W. Peofessob E. Ray Lankester,
British Museum (Nat. Hist.)
South Kensington, S.W. Downing Steeet,
nth June, 1901. The Under Secretary of
State for the Colonies presents
his compli-
ments
to the Under
Secretary for Foreign Affairs, and
is directed by tlie
Secretary of
State
to
transmit,
for
the information
of the Marquess of
Lansdowne, a copy of the
letter noted
in the subjoined schedule on the
subject of the Tsetse-fly and Buffalo. jNIwenga R.,
" Hook of the Kafue,"
N.E. Rhodesia,
24:th February, 1901. Mr. Val Gielgud to the British South African Company. -'
Sir,—I see from the papers
tliat
the
international conference for
the
preservation of
biir
irame
has decided
to recommend that buffalo be
L 2 L 2 First Report on
Ecoiioinic Zoology. 148 placed on the
list of game to be absolutely protected. I, therefore, wish
to bring
to your notice my observations on the Tsetse-fly and the Buffalo,
the
correctness
of which
are,
I
believe, borne out by the
experience
of
Mr. George Gray when
travelling through a fly country in 1899 and
also
by information obtained from native sources. The
Tsetse-fly has always abounded
in
districts where
Buffalo were
numerous, and
since the almost
total destruction of Buffalo by rinderpest
these
flies have not disappeared, although, perhaps, not
so numerous
as
formerly. The bite of the insect, however, appears
to have become much
less deadly to domestic animals, and
stock and dogs not only survive
the
bites for a much longer period than formerly, but in many cases suffer no
ill at all. This has been my personal
experience, and
I think
I am correct
in
saying Mr. Gray's is similar. British Museum (Nat. Hist.),
Cromwell Road, London, S.W.,
24«i June, 1901. British Museum (Nat. Hist.),
Cromwell Road, London, S.W.,
24«i June, 1901. To Sir Clement Ll. Hill, K.C.M.G., C.B. To Sir Clement Ll. Hill, K.C.M.G., C.B. 1. TSETSE-FLY AND BUFFALO CORRESrONDEXCE. The natives say that now the Buffalo are dead
the Tsetse-fly no longer
kills stock, but of course natives are notoriously careless and loose in their
statements. It
is
a
fact
also that
in many
places where
Buffalo
have
become extinct the Tsetse-fly has also vanished
; this
is the case in parts of
Sebungwi and Zankie districts. As there are large fly districts in northern
Rhodesia I call your attention to these facts, as
it appears to me that the
protection
of
the
Buffalo and
the
Tsetse-fly are
identical, and
I would
doubt
if the
benefit accruing from
the
preservation
of the former will
compensate for the disadvantages arising from the existence of the latter. I am, etc.,
Val Gielgud. I am, etc., Val Gielgud. To Sir Clement Ll. Hill, K.C.M.G., C.B. Sir,—I have the honour
to acknowledge the receipt of your letter
of
the
15th
instant, enclosing
a copy
of
a
letter from Mr. Val Gielgud
with reference to Tsetse-fly and Buffalo. In accordance with your request that I should furnish the Marquess of
Tjansdowne with some
observations on the subject I would
wish, first of
all, to draw your
attention
to
the powers given by Article IV. of
the
"dispositions" adopted by the
Conference
of
Plenipotentiaries
on
the
preservation of African wild
animals, May 1st, 1900. The final clause of
that
article was inserted on my suggestion with a view
to such a
case
as
that
reported
by
Mr. Gielgud, and
gives
power
to dispense
with
the
principles agreed upon " dans un interet superieur d'administration." It is, therefore, within the provisions of the agreement signed by
the
Plenipotentiaries for the Government to authorise the British South Africa
Company to destroy Buffalo, in order to
protect domesticated
cattle from
disease, A question, however, of a very serious nature arises as to whether there
is sufficient ground
for concluding
that
the
parasite
of the blood, which
is introduced by the
bite
of the
Tsetse-fly into domesticated animals, is
specially and ahundantly harboured in the blood of the wild Buffalo. The
theory
is held that
the
parasite {Herpetomonas nagance)
of
the
blood is comparatively harmless
to wild indigenous forms, such as Buffalo, Reports
to the Foreign
Office. 149 Antelope, etc., although multiplying in their blood, but that
it
is deadly to
introduced domesticated animals. Hence,
it
is supposed,
it
flourishes
in
the wild game and
is more abundant among them
tlian it would be if
its
presence caused death. I am inclined to believe this theory correct, but it
has not been properly tested. Before the destruction of Buffalo
in the vicinity of herds of domesti-
cated cattle is authorised,
it ought
to be clearly shoivn by experiment and
observation of competent
medical men
that
the
Buffalo harbours
the
parasite, or at any rate that it can harbour it without
being
killed off
as
are
domesticated
animals. It would
be
a
matter
of small expense,
in
comparison
with
the
enormous
pecuniary
interests
involved,
for
the
British South
Africa Company to employ a medical authority to experi-
ment on wild Buffalo, captured and kept
in a paddock for the purpose of
settling the question. To Sir Clement Ll. Hill, K.C.M.G., C.B. And it seems to me that authority to destroy the Buffalo should not be
granted to the Company until they have furnished
satisfactory scientific
evidence of the harbouring of the Nagana parasite by the Buffalo. I am. Sir, Your most obedient humble servant, Your most obedient humble servant, Your most obedient humble servant,
(Signed)
E. Ray Laxkestek. (Signed)
E. Ray Laxkestek. FoBEiGN Office,
15th October, 1901. SiE,—With reference to your letter of the 24th June, I am directed
by the Secretary of
State for Foreign Affairs
to
transmit to you for your
information the accompanying
copy
of
a despatch from
His
Majesty's
Commissioner
in the East Africa Protectorate
relative
to the connexion
between Tsetse-fly and the Buffalo. SiE,—With reference to your letter of the 24th June, I am directed
by the Secretary of
State for Foreign Affairs
to
transmit to you for your
information the accompanying
copy
of
a despatch from
His
Majesty's
Commissioner
in the East Africa Protectorate
relative
to the connexion
between Tsetse-fly and the Buffalo. I am. Sir, I am. Sir, Your most obedient humble servant, Your most obedient humble servant,
(Signed)
Maetin Gos.selin. Maetin Gos.selin. Maetin Gos.selin. To Sir Charles Eliot, K.C.M.G., C.B.,
His Majesty's Commissioner and To Sir Charles Eliot, K.C.M.G., C.B.,
His Majesty's Commissioner and Consul General, Mombasa. To Sir Charles Eliot, K.C.M.G., C.B.,
His Majesty's Commissioner and Consul General, Mombasa. His Majesty's Commissioner and Consul General, Mombasa. Sir,—I beg to acknowledge receipt of a copy of the correspondence
re
Tsetse-fly and
the preservation of
the Buffalo which you forwarded
to
uie for my opinion, and I have the honom* to transmit
to you herewith my
observations on the subject. The
Tsetse-tly belt
of
British East Africa, exclusive of Jubaland, may
be said to extend from Mtoto Andes to Simha, a distance of roughly ninety
miles
;
it
is
situated
in a densely wooded, low-lying
part
of the country,
about 3000 feet to 3400 feet above sea level. Driajani, an old camping ground, within
this
area, was considered by
the late Captain Haslam and myself to be the most dangerous place for fly,
on the old
transport route, but strange
to say it was
practically devoid of
game of any kind. In my opinion, Buffalo and other big game are not the only factors in
the Tsetse-fly theory, and we must
first consider the question
of climate
and humidity before we condemn the Bos caffa as the true and only source
of the Tsetse-fly and Tsetse-fly disease. I believe
that the distribution
of the
fly
is
entirely influenced by the
]>liysical
aspects of the country and
that for
its existence
it must have a
humid, low-lying position. Major Bruce
in
his
excellent report says
(see
p. 20. Professor E. Eay Lankestee. Mombasa,
&th September, 1901. To the Marquess of Lansdowne, K.G., etc., etc. To the Marquess of Lansdowne, K.G., etc., etc. To the Marquess of Lansdowne,
.G.,
,
My Lord,—In
reply
to Your
Lordship's
despatch, No. 259
of
July
20th,
respecting
the
connection
between
the
Tsetse-fly
and
the
Buffalo, I have
the honour
to
transmit
letters from
Messrs. Stordy and
MacClellan and Doctor Radford. After reading
this correspondence and
discussing
the
question with
other persons, my own opinion is that where there are Buffaloes, Tsetse-flies
are usually (but not always) found, but that the flies also occur in districts
where
there
are no
Buffaloes. Hence
it would
appear that the Buffalo
cannot be the only host of the parasite which the Tsetse-fly introduces into
the blood of domestic animals with fatal results. I have, etc., I have, etc.,
(Signed)
C. Eliot. C. Eliot. (Signed) First Report on Econo/nic Zoology. 1 50 Nairobi,
East Africa Protectorate,
'drd September, 1901. Nairobi,
East Africa Protectorate,
3rd September, 1901. Nairobi,
East Africa Protectorate,
3rd September, 1901. To R. SxoRm-, Esq., M.R.C.V.S. To R. SxoRm-, Esq., M.R.C.V.S. To R. SxoRm-, Esq., M.R.C.V.S. Dear Mr. Stordy,—^Many thanks for sending me the communi-
<iations
you have received
from
Sir
Charles
Eliot,
re
the
Buffalo and
Tsetse-fly. That the
two should
be
associated
is
not
extraordinary when
one
remembers
that both
inhabit
densely-wooded, damp, secluded
districts,
but
the arguments advanced
to prove that
the Bos
Gaffa alone
is
the
host of the Tsetse, and
that the extermination of the former leads to the
disappearance of the latter do not appear to me to be convincing. From
my own observation I am inclined to the opinion that hosts other than the
one
species
mentioned
(or
some
other
factors)
are
necessary
for
the
propagation
of the
fly, and that the hsematozoou
is in
all probability to
be found in many species of diptera. During my residence
in
Jubaland, East
Africa
Protectorate, which
•extended over two
years, I had ample opportunity of studying
the habits
and distribution
of
the
Tsetse-fly and the
effect
it produced on domestic
animals. Speaking
generally,
the
fly
belt
is confined
to two
distinct
areas
in
that province. (1) The
valley
of the Juba River within
the
forest
belt. Here
fly
abounds
for
a
distance
of
upwards
of
400
miles—in
fact,
so
general
is
it in places that
it
is a source of annoyance to Europeans and natives. Yet within the whole
of that great
tract of country Buffalo are few and
far between. The late Mr. Jenner and
I came
across them in one place
only (Lake Ohdey and district). Within
this
belt
is the district of Gosha,
80 to 100 miles in length, where the fly is peculiarly abundant, yet Buffalo
are not found
;
notwithstanding
this,
the ha3matozoon
is very
virulent,
and on the occasion of the late Mr. Jenner's expedition to Lugh in
189'J,
he
lost
every camel
and
pack-ox
that went
through
{vide my report,
May 16th, 1899, forwarded
to
the Secretary of State for Foreign Affairs,
No. 53, May 31st, 1899). y
(2) The region
of Lake Kumbi
to the
north-east
of Desek Wama
(Lake Hardinge), where the
distribution
of
the
fly
is
restricted
to the
dense
forest belt
in the neighbourhood
; yet
Buffalo
are not known
to
frequent
this
district. To Sir Charles Eliot, K.C.M.G., C.B.,
His Majesty's Commissioner and Further report
on Tsetse-fly
disease
in
Zululand,
1896),
"That
the
presence
of wild
•animals
in the
vicinity of horses and oxen
is not the
01dy factor in the
problem
is shown by the fact that
in
the
old days when
big game was
numerous
and roamed
over
the
whole
country,
hunters
and
travellers
never complained
of
fly
until they encountered
the
disease
in low-lying
tracts of country or along the
large river valleys." As in the Hermansdorp district
of Cape Colony herds
of Buffalo
are
still
to be found, yet
Tsetse-fly with its concomitant
disease
is unknown,
so in the high
altitude
of the Kedong
(6000
feet), in
this
Protectorate,
herds of Buffalo are to be met with, greatly reduced in numbers by rinder-
pest within recent
years
it
is
true, yet neither
Tsetse-fly nor
Tsetse-fly
•disease have ever been known to occur, nor has
the fly or its disease been
heard of in the Baringo district of the Uganda Protectorate, where herds
iii Buffalo and other big game exist. g
When studying
the causes which rendered
the
Island
of Mombasa
uninhabitable for
horses, I ascertained that an organism, the morphology
of Avhich was identical with that found in animals suffering from Tsetse-fly
disease, was found in donkeys which had never left the island. I expressed an opinion then {vide Preliminary Eeport as
to the causes
which rendered the Island of Mombasa uninhabitable
for horses in IM)',))
with regard
to African Nagana and Indian Surra being one and
the same
disease, and as the occurrence of Surra cannot be attributed to the presence
of wild animals or
Tsetse-fly, we mast
explain, ere we destroy the
buffalo
in an attempt
to stamp out
Nagana, why a
disease
identical
with
that Reports
to the Foreign
Office. 151 calised by
the
bite
of
the
Glossina
morsitans
occurs
in
places such
as
Mombasa, where Tsetse-fly and Buffalo are non-existent, calised by
the
bite
of
the
Glossina
morsitans
occurs
in
places such
as
Mombasa, where Tsetse-fly and Buffalo are non-existent, I have, etc., gned)
Robert J. Stordy, M.E.C.V.S. (Vety. Officer, E.A.P.). (Signed)
Robert J. Stordy, M.E.C.V.S. (Vety. Officer, E.A.P.). (Signed)
Robert J. Stordy, M.E.C.V.S. (Vety. Officer, E.A.P.). To R. SxoRm-, Esq., M.R.C.V.S. It appears
to me reasonable to assume that where
fly is abundantly found extending over a large
tract
of country that
its
natural host
(if one only) should be
in large numbers also
;
or else the fly
must be possessed of extraordinary migratory powers. y
g
y p
If
that
host
be
the
Buffalo,
it
is
strange
that
it
is
particularly
conspicuous
by
its absence
in
the
extensive
districts mentioned,
while
waterbuck and bushbuck
are common
in most
parts, and yet
again
in First Report on Economic Zoology. 152 others
no
animals
of
any
sort
are
found
save
monkeys
and
rodents. Amongst
other
pests that
affect camels very
seriously
in Jubaland
is a
species of gadfly which
is restricted
in
its distribution
to the open plains
and sparsely-bushed country
in the
districts
of Desek Wama and IJerib. This
fly, unlike the
Tsetse, attacks animals during the day
at
all hours,
and
the
symptoms
produced
by
it
(in
camels)
are
identical
in
every
particular with those produced by Tsetse, but I was unable at the time to
substantiate
this
theory microscopically owing
to lack
of
the
necessary
materials, etc., for carrying on investigations. Yours truly, William
S. Radfobd, Medical Officer, East Africa Protectorate, Nairobi,
ith September, 1901. Sm,—Eeferring to our conversation on the subject of Tsetse-fly in
the Jubaland Province, I would
state that along the Juba Eiver where
low-lying forest exists (my observations cover a distance of some hundred
and twenty miles from Bulbula
to Gele)
Tsetse-fly abounds. The worst
places are damp, dark, and low-lying, shaded
chiefly by the Ndoma Palm. In many
such
localities
there
is
little
or
no game
and
certainly no
Buffalo. On
the
other
hand,
from
Mtudo
northwards
buffalo
are
found,
especially
in
rainy weather
and
in
the
heat
of
the
day frequent
the
thickest depth of the forest
;
here, too. Tsetse-fly is found in large numbers,
as also in the dry lake beds adjacent to the river. At the same time half
a mile
or
so away from the
river and lake beds
in the dense dry bush,,
Avhere
Buffalo
feed
in
the
early mornings
and
evenings,
the
fly does
not appear. Reports
to the Foreign
Office. Reports
to the Foreign
Office. 153 Majesty's
Commissioner
in
the
British
Central
Africa
Protectorate
respecting the supposed connexion between Tsetse-fly and I>uffalo. I am, etc, Clemext
Ll. Hill. Clemext
Ll. Hill. To The Dikector,
Natural Historv Museum. The Eesidbncy, Zomba,
British Central Africa Protectorate,
mth Septeinber, 1901. To R. SxoRm-, Esq., M.R.C.V.S. In many places the areas
in which
fly exist are quite smalU
possibly only a few hundred yards in
extent, and
I have seen a watering
])lace made entirely free of
fly for the time being by cutting down forest
and
undergrowth
and
burning
the
grass
in
the
immediate
vicinity. Again,
at
Lake
Hardinge
(now
dry), where
little
or no
forest
exists,
where buffalo
are frequently
seen, and always
large herds
of waterbuck,
and Somalis graze their cattle
at
all times
of the year without
ill effects. Passing
on, however,
some
thirty
miles
to
the Rumbi
forest
on
the
Affmadu road. Tsetse-fly abounds, especially in wet weather. My opinion, therefore, is that
Tsetse-fly is
to be found in certain dark,
damp, low-lying localities, irrespective of big game of any kind. I have, etc.,
(Signed)
J. W. P. McClellan. Foreign Office,
^Oth November, 1901. ,
Sir,—With
reference
to my letter
of
October
15th
last,
I am
directed by the Secretary of State for Foreign Affairs to transmit
to you
for
your information
the accompanying
copy
of
a despatch
from
His To The Maequess of Lansdowne, K.G. To The Maequess of Lansdowne, K.G. My Loed,—I have the honour to acknowledge the receipt of your
Lordship's despatch No. 190
of July 20th, with enclosures regarding
the
question
of the existence of the
Tsetse-fly in connection with the
preser-
vation of the Buffalo, and in reply to submit the following remarks, in so
far as my own experience has taught me, on this vexed question. I may say at once that
I am lirmly of the opinion that
in East Africa
the
existence
of the
Tsetse-fly was never in any way connected with
the
presence of the Buffalo more than any other species of game. I first met with the ti'ue Tsetse, in any great numbers, and consequently
suffered much from
their
needle-like
bite, in German East Africa, about
eighty miles inland from Saadani, in February, 1886. At that time impala,
hartebeest, zebras, and warthogs were found
in
large numbers,
also
a few
sable
antelopes, but
there were no
Buffaloes
anywhere in the vicinity of my shooting grounds. y
In 1887
I again found
this
fly in
great numbers
in a small patch of
thick bush, about a mile and a half long and three quarters of a mile wide,
about ten miles west of Taveita, In this bush which projected from the forest I certainly found buffaloes
occasionally, but as a rule they preferred
to lie up for the day in the thick
and cooler forest, in which there were no Tsetse-flies. The bush
in question was a favourite
resort
of impalas, and a small
dik-dik (Modoqua), the latter in great numbers, and also a few bush-bucks
and waterbuck. At that time (1887) Buffaloes may be said to have swarmed
in
the
vicinity
of
Tareita, but
I
never
saw
a
Tsetse-fly
in
this
one
particular patch of bush. Later
on, iu
1888-89
and
1890, the
fly was met
with, also
in great
numbers, along the old caravan road from about two miles south of the
Tsavo river, as far as Kibwezi. Between these two points there were practi-
cally no Buffalo, but a great number of dik-dik and a few impala. The
flies and the small game are still there, but there are certainly no Buffaloes. In 1891-2, after rinderpest had carried
oft" nearly all the Buffaloes (at
least 90 per
cent.) throughout East Africa, Mr. The Eesidbncy, Zomba,
British Central Africa Protectorate,
mth Septeinber, 1901. To
His
Majesty's
J^kincipal
Seceetary
of
State
for
Foreign
Affairs. To
His
Majesty's
J^kincipal
Seceetary
of
State
for
Foreign
Affairs. My Lord,—With
reference
to
your
Lordship's
despatches Nos. 141
and
155,
enclosing copies
of
correspondence on the subject
of the
connection between
the
existence
of
Tsetse-fly and
the
preservation
of
Buffalo, I
have
the
honour
to append a few notes which
give my own
experience during the past fourteen years in Africa on this subject. (1)
Tsetse-fly
would
appear
to
depend
upon
wild game
for
their
existence,
as
I have never found Tsetse
in any locality where game was
totally non-existent. (2)
Tsetse does not appear to be in any way specially dependent upon
buffalo. On the
plains
at the north end
of
Nyassa,
before
rinderpest
made
its
appearance,
there were
vast herds
of
Buffalo, but
no
Tsetse. The natives at the north end at that time owned large quantities of cattle
which
could
be
seen
grazing
in
close
proximity
to
Buffalo. When
rinderpest came it killed practically all the cattle and all the Buffalo. In
other districts
of
British Central Africa Tsetse are found
in large
quantities where Buffalo, at the present date, at any rate, do not exist. (3)
Tsetse are not found
(in
British Central Africa)
in open
plains,
although such
plains may have
large
quantities
of game on them, and
in
spite of the fact that
at the edges of the plains, where forest abounds
Tsetse are found. It would appear, therefore, that what regulates the presence of Tsetse-
fly
is the description
of the country almost as much
as the abundance or
scarcity of game. I have, etc.,
(Signed) I have, etc.,
(Signed)
Alfred Sharpe,
His Majesty's
Commissioner and Consul- General. I have, etc.,
(Signed)
Alfred Sharpe,
His Majesty's
Commissioner and Consul- General. (
g
)
p
His Majesty's
Commissioner and Consul- General. Foreign Office,
21th November, 1901. Foreign Office,
21th November, 1901. To The Director, Natural History Museum. Sir,—With reference to my letter of the 20th instant, I am directed
by the Secretary of
State for Foreign Affairs to transmit
to you for your
information a copy of a despatch which has been received from the acting
British
Commissioner
in
Uganda,
respecting
the
supposed
connection
between Tsetse-fly and Buffalo. I am, etc., Clement
Ll. Hill. First Report on Economic Zoology. 154 Entebbe, Uganda,
TitU Seiitanhcr, lUOl. To The Maequess of Lansdowne, K.G. Rogers, the present sub-
commissioner
of the Tanaland province, and myself found
the
Tsetse-fly
existing in considerable numbers
in a narrow belt of forest, not more than
a mile wide, between Mkonumbi and Witu, and we were told by the natives
that the
Gallas, when driving
cattle to Lamu for
sale, always drove them
through
the
forest
by
night, and
that
the
herdsmen
carried
smoking
firebrands to keep the
flies off. p
With the exception of a few bushbnck and duykers, there was no game
in the vicinity of this belt of forest. These
four
places
are
the
only
areas,
the
first and
third
ones only
being
of any considerable
extent, in which
I have myself met with the
true Tsetse-fly, and yet, until they were decimated by rinderpest. Buffaloes
were more
or
less common throughout East
Africa, and
perhaps
in no
part
of
the
Continent were
they ever more
plentiful
than
the
Masai
country
between
Kilimanjaro
and
Lake
Baringo, Man
Plateau,
and
Turkwell. Throughout the whole of
this vast area the Tsetse was, and
is,
non-existent. Reports
to the Foreign
Office. Reports
to the Foreign
Office. 155 I may add that
there
is a species of
Tsetse-fly found along the wooded
portion of the lake shore here at Entebbe. I may add that
there
is a species of
Tsetse-fly found along the wooded
portion of the lake shore here at Entebbe. A specimen
of
this fly I gave to Sir Harry Johnston, and I
believe he
sent
it home. It
is plentiful in the botanical gardens. In these gardens,
with
the
exception
of
a few monkeys and
squirrels, and
certain small
nocturnal
beasts,
such
as
the
ichneumons,
etc.,
and
an
occasional
hippopotamus,
there
are
no mammals, and
if, as
is supposed, the
fly
is
necessarily dependent on the presence
of
suitable mammals on which
to
feed, the blood
of these animals, and
occasionally man, must
necessarily
form
its food supply. In conclusion I may add that
I have ventured
to
hold
the opinion that the Tsetse
is
like the mosquito, only a bloodsuckei-
by predilection, and, in support of this view, I may state that on my return
to Kibwezi in April, 1892, at a time when the whole of the fly "belt" was
parched and dried up—there being no water between Msongoleni and the
Tsavo river, a distance of fifty miles
; and consequently there was no game
of any kind—the Tsetse was more plentiful than at any other time, before
or since, I have passed through that area. Between Mtoto-Ndai and Kinani
I caught on my own person thirteen
of these
flies, and my half-naked
porters
su-ffered even more than I did
from their bites. I
can, therefore, not
readily believe
that
all
these
flies could
exist
in
such a dried-up and at that time intensely hot
locality if solely dependent
on the blood of a very infrequent passer-by or a stray dik-dik, I have, etc.,
(Signed)
F, J. Jackson'. I have, etc.,
(Signed) I have, etc., (Signed)
F, J. Jackson'. F, J. Jackson'. Sudan Government,
Civil Secretary's Office, Cairo,
1th August, I'JOl. Sudan Government,
Civil Secretary's Office, Cairo,
1th August, I'JOl. To TiiK Beitish Ageijt and Consul-General, Cairo. Sir,—We are much troubled
in the Sudan by White Ants. They
destroy not only wooden
telegraph
poles,
boxes,
furniture,
timber,
etc.,
but in the Khartoum district green and growing plants. This
is in our experience an unusual procedure for the Sudanese White
Ants (who mostly confine themselves
to wood), and shows that there must
be several varieties
of the
pest. This particular form
of White Ant has
its nest about the size of a small melon, 4 or 5 feet under ground
;
but
it
is
very
diiflcult
to
extirpate
him
completely without
digging
up
and
spoiling a great deal of ground. First Report on Economic Zoology. 156 As Lower Egypt is not troubled by these insects, I have applied in vain
to the School of xYgricultiire at Cairo for information as to the best method
of getting rid of them. I may add that we have planted a good number
of Casuarina trees, which are supposed
to be proof against the attacks of
AVhite Ants
; but they eat the trees with the greatest impartiality. I have the honour, therefore, to suggest that you will have the kindness
to forward a copy of
this
letter to the proper quarters with a request that
I may be supplied with any information there may be on the subject, or
that I may be referred to any books or papers on the same. I am, etc., I am, etc.,
(Signed)
Gleichen, Majoe,
Assistant Civil Secretary for Governor-General. Cairo, Wi August, 1901. To The Maequess of Landsdowne, K.G., etc., etc. To The Maequess of Landsdowne, K.G., etc., etc. My Lord,—I
have
the
honour
to
transmit
to
your
Lordship
herewith copies of a note which
I have received from the Civil Secretary
to the Soudan Government, asking for assistance
in procuring information
as to the best means for combating the ravages of the White Ant, which
is
extremely destructive in the Soudan. I am informed that, in certain parts of America, the White Ant is very
prevalent, and that considerable attention has been directed to this subject
by the Department
of Agriculture
in the United
States. It
is prol)able
also that the Colonial Office are in possession of valuable information, and
more might perhaps be obtained from the Horticultural Gardens at Kew. Reports
to the Foreign
Office, 157 Sudan Government,
Civil Secretary's Office, Cairo,
1th August, I'JOl. I
should
be
most
grateful
if
your
Lordship would render me any
assistance which
is
possible,
in
obtaining
such
information as muy
l)e
available, for the use of the Soudan Government. I have, etc., I have, etc., I have, etc.,
(Signed)
Eennell Rouu. (Signed)
Eennell Rouu. (Signed)
Eennell Rouu. FoBKiGN Office,
22nd August, 1901. FoBKiGN Office,
22nd August, 1901. To THE DiRECTOK OF THE ROYAL GARDENS, KeW. Sir,—I am directed by the Marquess of Landsdowne to transmit to
you the accompanying copy of a despatch from His Majesty's Acting Agent
and
Consul-General
in Egypt,
relative
to the ravages committed
in the
Sudan by the White Ant. I am to enquire whether the Director of the Eoyal Gardens can furjiish
any information on the best means
of
combating
the ravages
of
tliese
insects. I am, etc., T. H. Sanderson. Reports
to the Foreign
Office, Royal Botanic Gardens, Kew,
IZrd Angust, 1901. To Sir T. H. Sanderson, G.C.B., K.C.M.G.,
Foreign Office, Downing Street. To Sir T. H. Sanderson, G.C.B., K.C.M.G.,
Foreign Office, Downing Street. Sir,—I have the honour to acknowledge the receipt of your letter
of yesterday's date enclosing
a
copy
of
a
dispatch from His Majesty's
Acting
Agent
and
Consul-Genera I
in
Egypt
relative
to
the
ravages
committed in the Sudan by the White Ant. In reply I have to state that Kew is not in possession of anything but
the most general information on the subject and
is therefore unable to
furnish any advice which would
be
of any practical
utility to Sir Rennell
Rodd. I
have
forwarded
the
correspondence
to
the
Director
of
the
Natural History Museum, South Kensington, and requested him to examine
the question and communicate with you. I am, etc., I am, etc., I am, etc.,
(Signed)
W. T. Thiselton-Dyeu. I am, etc.,
(Signed)
W. T. Thiselton-Dyeu. (Signed)
W. T. Thiselton-Dyeu. British Museum (Nafc. Hist.),
Cromwell Road, S.W.,
Itli September, 1901. British Museum (Nafc. Hist.),
Cromwell Road, S.W.,
Itli September, 1901. British Museum (Nafc. Hist.),
Cromwell Road, S.W.,
Itli September, 1901. To Sir T. H. Sanderson, G.C.B.,
Foreign Office, S.W. To Sir T. H. Sanderson, G.C.B.,
Foreign Office, S.W. Sir,—I am directed by
Professor Eay Lankester
to acknowledge
the receipt of your letter of the 22nd ult. addressed to the Director of the
Royal Gardens, Kew, enclosing copy of a despatch.from His Majesty's Acting
Agent and Consul-General
in Egypt, relative to the ravages committed in
the Sudan by the White Ant. I am to state, for the information of the Marquess of Lansdowne, that
Professor Ray Lankester
is giving
his attention to the question, and that
he will further communicate with you
in regard to the matter. I am, etc.,
(Signed)
0. E. Fagan. I am, etc., (Signed)
0. E. Fagan. British Museum (Nat. Hist.),
Cromwell Road, S.W.,
5th November, 1901. British Museum (Nat. Hist.),
Cromwell Road, S.W.,
5th November, 1901. Report on Termites or White Ants, and methods
of checking the ravages of the same, prepared
at the request of the Sudan Government. Without having
specimens of the White Ants
or Termites
that
arc
causing havoc
in the Sudan not only to wood-work, telegraph poles, etc.,
hut also to green crops
in the vicinity of Khartoum,
it
is not possible to
give
a
satisfactory account
of
any methods
for
combating
the
pests,
investigation
to
be
of
any
practical
use must
be made
on
the
spot. Information has been, however, collected from all sources concerning any
measures that have been taken in various parts of the world with a view
of
checking
the
serious damage
these
insects
do. This information
is
embodied
in
this report. The various African Termites are also einime-
rated—their various ways of working pointed out and detailed methods of
destroying them given. A number of suggestions for preserving
articles
from their attack and possible new remedies are also given. British Museum (Nat. Hist.),
Cromwell Road, S.W.,
5th November, 1901. To Sir T. H. Sanderson, G.C.B.. Foreign Office, S.W. Sir,—Referring to your
letter
of 22nd August
last to the Director
of the Royal Gardens, Kew, and
to my acknowledgment of
the 7th of
September, relative
to a despatch from His Majesty's Acting Agent and
Consul-General in Egypt on
the subject of the ravages committed in the
Sudan by the White Ant, I have
the honour to
enclose herewith, for the
information of the Marquess of Lansdowne, a report prepared by Mr. F. V. Theobald,
of
this Department, on the Termites
or White
Ants,
dealing
generally with their prevention and destruction. I am, etc.,
(Signed)
E. Ray Lankester. I am, etc., I am, etc.,
(Signed)
E. Ray Lankester. (Signed)
E. Ray Lankester. First Report on Economic Zoology. 158 Report on Termites or White Ants, and methods
of checking the ravages of the same, prepared
at the request of the Sudan Government. A coinplete list of African Termites is given in the Appendix (p. ISl). *
" Insect Life," II. 283. Damage caused by
Teraiites. Tlie usual way of working
is
to
destroy wood-work of
all
kinds. In
all
instances Termites work
in the dark
; they enter wood-work from the
ground, working up
inside the wood from where the
poles, supports and
timbers are placed
in the
soil. Furniture, books and papers are attacked
and
destroyed,
the wood-work
being completely hollowed
out, nothing
but a thin papery outer shell
left, which naturally can stand no
pressure
and so, soon collapses. Damage
to
living substances and crops
is by no means unusual. The
American
T. fiavipes
has
been
recorded
destroying
turnip
roots,
by
gradually eating out the
interior.*
In
Florida they damage living
trees
by eating away
the bark about the collar and
root, but growing wood
is
only attacked by them under exceptional circumstances when
there
is no
dead wood
or when
they wish
to
escape from
the
heated
soil.f
This
species
also
attacks
potatoes growing
in
rich
soil
or where
there
is
a
considerable quantity
of
decaying
vegetable
matter. The
insects form
scars or pits covering the surface, often over-hung by the dead and dying
skin. Termes fatih
is very destructive
to
trees
in Arabia.^
In Ceylon tea
and
coffee plants are attacked by them, the stems being gnawed through
just below the ground, Termes australis, according to French (" Handbook of Injurious Insects
of Victoria," pp. 11,
1?>7, 1893), attacks vines and fruit
trees
in
Victoria. ],)amage to living plants
is therefore not unusual. African Termites.* The following Termites
are common in Africa
; the species found
in
Central and Northern Africa being separately tabulated at the end of the
list. 1. Calotermes flavicolUs, T., Ent. S^\ st. Fabr., 11, 15,
\\. 91. 2. Hodotermes ocliraceiis, Burm. =
Termes ocJiraceus, Ramb., Egyp. Xeurop., pi. 2,
fig. 21, Uaiiib. =
Termes ocJiraceus, Ramb., Egyp. Xeurop., pi. 2,
fig. 21, Uaiiib. o. Hodotermes mossavihicus, Hagen, Linn. Ent., 12, p. 94. o. Hodotermes mossavihicus, Hagen, Linn. Ent., 12, p. 94. 4. Termes heUicosus, Smeathman,
Pliil. Trans., Vol, 71, p. 141. 5. Termes ang^istatus, Eamb., Neuropt, p. 306, No. 11. 6. Termes capensis, De Geer, Memoirs, VII., p. 47, tab. 38,
fij;. 7. 5. Termes ang^istatus, Eamb., Neuropt, p. 306, No. 11. 5. Termes ang^istatus, Eamb., Neuropt, p. 306, No. 11. 6. Termes capensis, De Geer, Memoirs, VII., p. 47, tab. 38,
fij;. 7. 6. Termes capensis, De Geer, Memoirs, VII., p. 47, tab. 38,
fij;. 7 7. Termes destructor, Smeathman, Phil. Trans., Vol. 71, p. 141, No. 4, tab. 10. 8. Termes ludfugus, Kossi, Mant. Etr., 1, p. 107. 9. Termes {Eutermes) atrox, Smeathman, Phil. Trans., Vol. 71. 10. Termes (Eutermes) lateralis, AValker, Lin. Ent., 12,
}). 216. 11. Termes (Eutermes) trinervius, Eamb., Ntriiropt., p. 308. 12. Termes (Etitermes) mordax, Smeathman, Phil. I'rans., Vol. 71, p. 141. 12. Termes (Etitermes) mordax, Smeathman, Phil. I'rans., Vol. 71, p. 141. 13. Ttrmes viator, Lat., Hist. Nat., XIII., p. 51, 8. 14. Termes {Eutermes') arhorum, Smeathman, Phil. Trans., Vol. 71,
]>. 141. 15. Termes incertas, Hagen., Linn. Ent., 12, p. 230. .Species found in N. Africa, Egypt, Sudan, etc., down to the Equator. .Species found in N. Africa, Egypt, Sudan, etc., down to the Equator. p
The following
species occur
in North and Central
Africa and along
the seaboard
: C. flavicolUs
)
. i Algeria. us
)
°
C. lucifug
T. trinervius, Tripoli
C. flavicolUs
I
T. atrox
T. ludfugus
JT. ochraceus
Egypt. Reports
to the Foreign
Office. 159 T. heJ/icosus
\
I^o'ig'^1^'' I^^rf^i'-. KordofMU,
I
Sennaar, and Abyssinia. T. destructnr, Kordofan. IT. viarum
T. helUcosus
T. destructor
2\ mordax
\
T atrox
I
^^^^'^'^ Leone and Gambia. 7\ arhorum
T. trinervius
T. lateralis
I
T.fatale, Arabia. X "History of Arabia, Ancient and Modern," A. Crichton, 1833. *
" Insect Life," II. 283.
t
" Insect Life," I. 341.
X "History of Arabia, Ancient and Modern," A. Crichton, 1833. t
" Insect Life," I. 341. *
" Insect Life," II. 283.
t
" Insect Life," I. 341.
X "History of Arabia, Ancient and Modern," A. Crichton, 1833. A'abieties of Nests
{Termitaria). Termites
or White Ants form
variously-shaped
nests. The ways of
destroying Termites
differ
according
to
the
type
of
Termitaria. The
following types of
nests seem to occur (1) large mound-nests, often six to
ten feet high {T. beUicosus), (2) small dome-shaped nests over tree stumps,
seldom more
than two
feet high {Eutermes
sp.),
(;>) Arboreal
nests, on
live and dead
trees, approached by a covered
tunnel up
the
tree trunk
(E/ftermes arhorum and Eutermes sp.),
(4)
Small round
nests
in
the
soil
mentioned by Major Count Gleichen in his letter of inquiry (sp. ?). First Report on Economic Zoology. i6o Torinitaria of the arboreal species are also found on the roofs of houses,
stables,
etc. The
nests occur both above and below ground. Methods
of cstenniiiation must therefore depend on the species causing destruction. Methods of Peevention and Remedies. As the White Ants nearly always work under cover, the damage they
do is often not detected until too late. It is therefore necessary to protect
objects from being attacked. This can be done (1) by making ground
wood-work either obnoxious, poisonous or inaccessible to the Ants
;
(2) by
lessening the number of Termites by poisoning and destruction of their
nests. * "Insect Life,"
I. 35B. Destruction and Prevention where damaging living Trees. When
the Termites
attack
trees by eating away
the bark about the
collar and
root,
the
earth
should
be removed from
the
infected
parts
and
the
ground should be
exposed
to the depth
of
several
inches, and
the dead wood and bark cut
off with a
knife. A
liberal
supply
of
hot
water will destroy those that cannot be reached with a knife. Pyrethrum
and
kerosene enudsion
in
extreme
dilute
solution can be
applied with
success, but the
latter should be
used with great
caution. Trees which
have been
girdled may
be
saved by
inserting
scions
between
the
root
below and
the stalk
above,
thus
re-establishing
the
connection between
the two. A
poultice of mud and cow-dung applied to
the affected part
will protect it and assist in the formation of new bark.f Destructiox of Termites and their Nests. When the nests can be
located much good can be done by destroying
the
nests and
inmates. This may be done by pouring
kerosene
oil
or
carbolic into
the
nests. The
action
is temporary,
however,
for
it
only
•drives many of the ants away to form fresh nests. Clearing White Ants from Wood-work. When present
in
wood-work
of
a house
or
other
building,
Riley *
suggests
injecting steam
or hot water
or kerosene wherever an opening
seems to lead into their burrows in timber. Use of Arsenic Poisons for the
Insects. A
far better plan
is
to put
arsenic and syrup
into
the openings
of
their tunnels
or
into the
nests. The
arsenic may be mixed
up
with
sugar
into a thick syrup. Paris green would probably answer well. Tlie
insects
will feed
off
this and soon die, and
it
has
been found
that
the
dead
Termites
are devoured by other Termites which themselves become
poisoned, and
so
great numbers
are
destroyed. Experiments
should
be
made
in
this
direction and if successful should
be carried out on a
large
scale. "Where this plan
has been tried
in
isolated
nests
it has met with
"marked success. First Report on Economic Zoology. First Report on Economic Zoology. 1 62 Making Ground Wood-work Obnoxious and Poisonous
TO Termites. Steeping
posts,
poles,
timbers
of
houses,
etc.,
\\\
various
smelling
substances has met with more or less success. Use
of Creosote.—In
India creosote was used by the Government for
treating all the railway sleepers before they were laid. In the
outskirts of Columbia great damage has been done by White
Ants (T. Jfavipes) to board fences
:
" The chief damage is done where the
boards meet on the posts. It
is
particularly noticeable where a batten
is
nailed on
at
a
joint. Professor Atkinson
states
that
tar
poured
on
between the posts and
the boards soon after building the fence will act as
a preventive." * Experiments conducted with wood-boring
insects and
creosote-soaked
posts has not
invariably met with success with Termites
or
other
insects
after
the wood has been " planted " some
time. It cannot
therefore
be
recommended
for telegraph
poles,
etc.,
that
are
being
destroyed
in
the
Sudan. Use of Arsenioi/s Soda.—Of. more lasting effect is steeping the parts of
poles, etc., that
are
placed
below ground
in
arsenious soda
dissolved
in
mineral oil. Protection
of Telegraph Poles and Buildings.— Telegraph poles might
easily be protected by having the part buried
either embedded in cement
or encased in zinc or tin. The metal should be painted with non-corrosive
paint. Cement casing would be
best, as the
tin would probably corrode
rapidly unless carefully painted, and the least hole w^ould let the pests into
the wood. Zinc
casing has been employed for foundation wood-work
in
buildings with success, the zinc passing up the timbers out of the ground
and then bent over (Fig. 17, 2) so as to prevent the ants crawling upwards. Complete dryness in buildings
is also essential
in regard
to checking some
species of White Ants. All floors of houses
in the districts where White
Ants
are
destructive
should
be made
of
concrete (or
raised
well
above
ground,
as shown
in
Fig. 17,
3). No
furniture
should be allowed
to
stand
against a
wall. Where wooden
floors
are
essential, the
furniturr
may be
protected by standing the
legs
in
small tins with
paraffin
oil
in
them. * "Insect Life,"
I. 35B. Fig. 2. \vrr«^
a
:r-3
•
•
•
'c-
•. Fig.l. \vrr«^
a
:r-3
•
•
•
'c-
•. Fig.l. Fig. 2. •
'c-
•. Fig.l. t Riley, " Insect Life,"
I. 341. *
" Insect Life," II. 253.
t Riley, " Insect Life,"
I. 341.
X Bull. 30 (N.s.), Dept. Agric, U.S.A. *
" Insect Life," II. 253. Reports
to the Foreign
Office. Reports
to the Foreign
Office. 163 To The Civil Secretary,
Sudan Government, Cairo. Dear
Sir,—Referring
to my letter of the 17th ultimo, I have to
inform you that a
full report on White Ants, dealing generally with their
prevention and destruction, was forwarded
to the Foreign
Office on the
5th
inst. to be transmitted
to His Majesty's Agent and Consul-General
in Cairo. I shall be glad to learn that it has reached you. I remain, etc., (Signed)
C. E. Fagan. (Signed)
C. E. Fagan. (Signed)
C. E. Fagan. Woods not attacked by White Ants. From a report concerning the ravages of the White Ant in St. Helena
it
is gathered
that
certain woods
resist the
attack
of these pests
better
than others. The wood of Myrtacexz and teak were found
to
be the last
attacked and
to
resist
their
ravages. A correspondent
in West
Africa
informs me
that they will not touch pitch-pine as mucli as other woods. Oalifornian red wood also appears to be free from attack.^ Conclusion. Beyond
these
points
nothing
is known
regarding
the
destruction
and prevention of Termites. It
is
certainly (1) advisable to
set
all foun-
dations of wood-work in cement so
as to prevent the entry of the
ants
;
(2)
to adopt
the
precaution
of
steeping wood-work
for
the ground
in
arsenious
soda, and (3) to employ arsenic
as
a poison
in the
nests near
all habitations, works and railways. The
probability
is that by such pre-
cautions the ravages of these pests in the .Sudan would be greatly lessened. (Signed) (Signed)
Fred. V. Theobald. Fred. V. Theobald. British Museum (Nat. Hist.),
Cromwell Road, S.W. \bUi November, 1901. \bUi November, 1901. To The Civil Secretary,
Sudan Government, Cai Sudan Government,
Civil Secretary's Office, Cairo,
24:th November, 1901. Dear
Sir,—I have the honour to acknowledge your letter of the
15th
instant and
to thank you for the " Report on White Ants," which
was received through H. B. M.'s Agency here. g
This report
will eventually be
of
the greatest
use
as a guidance
in
lighting these pests. I remain, etc., E. G. Blunt, Lieut.-Colonel. E. G. Blunt, Lieut.-Colonel. The Civil Secretary and Sudan Agent, Cairo. The Civil Secretary and Sudan Agent, Cairo. M
2 M
2 First Report on Economic Zoology. 164 COKRESPONDENCE AXB REPOPtT PREPAPtED FOR THE
FOREIGN OFFICE. Sudan Government,
Civil Secretary's Office, Cairo,
2ncZ October, 1901. To The Directoe,
Natural History Mnsfuni,
Cromwell Road, S.W. Bbitish Museum (Nat, Hist.),
CromweU Road, S.W. Bbitish Museum (Nat, Hist.),
CromweU Road, S.W. To The Civil Secretary,
Sudan Government, Cairo, Egypt. To The Civil Secretary,
Sudan Government, Cairo, Egypt. Dear
Sir,—Referring
to Count Gleichen's
letter of the 2nd inst. (No. C.S.S. 4/1259), I am desired by the Director to send you herewith a
report on the
subject
of
locust
plagues
in
the Sudan, with notes and
suggestions for the destruction of the locusts. I am
to point out that while Count Gleichen's letter of the 2nd
inst. refers
to locusts, his
letter
of the 7th August
is on the subject of White
Ants. A separate report will be forwarded
to you in due course in regard
to this last question, which
is forming the subject of investigation by the
Museum. I remain, etc.,
(Signed)
C. E. Fagan. I remain, etc.,
(Signed) (Signed)
C. E. Fagan. Report on Locust Plagues in the Sudan. At the request
of the Foreign
Office, the follo\nng information
regarding the Locust Plagues in the Sudan has been despatched for
the use of the Sudan Government. Particular attention
is
called
to
sections B. 2, 3, and 4 in the Report. Experiments should
certainly be
tried
in
connection with
the African
Locust fungus and the use of
*' poison-baits." Prevention and Remedies
for Locust Plagues. A. Destruction by capture
in,
1,
trenches
;
2,
traps
;
3, by burning,
This was fully dealt with in the proof of a jjaper sent from Egypt. A. Destruction by capture
in,
1,
trenches
;
2,
traps
;
3, by burning,
This was fully dealt with in the proof of a jjaper sent from Egypt. B. 1. Destruction of the eggs. 2. Collection of " hoppers " by special machines. 3. "Poison-baits." 4. Fungoid disease. g
5. Plants poisonous to locusts. 6. Natural enemies. Reports
to the Foreign
Office. 165 Appendix
(p. 179). Various locusts, North African. To The Directoe, Dear
Sir,—I wrote
to yon in August
last asking for such infor-
mation as you could kindly give me about the different species
of
locusts,
the means
of distinguishing them, and
their habits. I am sending you by
this mail a proof copy of the instructions
that have been drawn up for the
use of
oflBcers in the administration of the Sudan, in which
it
is proposed
to include your notes. Any remarks you might think
fit to make would be of great interest,
as the instructions will not be printed until your notes have been received
and included. These notes are being published with the shortest possible delay, and I
should
like
if possible
to have them circulated by the beginning of next
month. I should, therefore, be very much obliged
to you
if you could
let
me have your notes and remarks as soon as possible. I must apologise for the trouble
I am giving you, but the question of
the destruction
of
locusts
is
so important in
the Sudan that I feel
it my
duty
to
collect
as much information
as
I
possibly can before publishing
the notes. I remain, etc.,
(Signed) I remain, etc.,
(Signed) g
,
Assistant Civil Secretary and Sudan Agent, Cairo. British Museum (Nat. Hist.),
Cromwell Road, S.W.,
\Wi October, 1901.. To The Civil Secretary,
Sudan Government, Cairo. Sir,—I am desired by the Director to acknowledge receipt of Count
Gleichen's letter of the 2nd inst., enclosing proof of printed instructions for
the use of
officers in the administration
of the Sudan, with regard
to the
destruction of locusts. A full reply thereto will be sent as soon as possible. I have the honour to be, etc.,
(Signed)
C. E, Fa.g.\n, Assistant Secretary.. Reports
to the Foreign
Office. Various locusts, North African. Various locusts, North African. (J) By collecting. (J) By collecting. Egg masses may
be
collected where plenty of native
labour
can
be
obtained. The authorities in Cyprus in 1881 had 1300 tons of eggs collected by
natives at so much per pound. A look-out should be kept
to see where the females deposit their eggs,
and those particular parts of the district should be searched soon after. B.
1. Destruction of Eggs.
(a) By cultivation. The eggs are usually laid in firm ground to guard them against natural
enemies. By turning up and loosening the
soil
to a depth of three inches, the
eggs
can
be
exposed,
and
numbers
are
destroyed
by
birds,
parasitic
insects, etc. First Report on Economic Zoology. First Report on Economic Zoology. 1 66 B.
2. Machines for Catching
Locusts
(" Hopper Dozers "). These machines of various patterns resemble a shallow earth scoop or
long tray. They are largely employed in America in Locust plagues. A " hopper-dozer "
is
usually a flat iron or zinc tray, containing tar or
paraffin. This tray is dragged or pushed along by a horse or man against
the wind—the young locusts jumping out of the way get blown in, and are
thus killed. Fig. 18. a hopper-dozer used for COIiLECTING LOCUSTS. {After S. J. Hunter, Kansas). Fig. 18. a hopper-dozer used for COIiLECTING LOCUSTS. {After S. J. Hunter, Kansas). One machine mentioned
in "Insect Life"
is 15 feet long, 2 feet deep,
and 4 to 5 feet wide
;
this box
is divided into sixteen compartments filled
with lime water. A plan of one of the most recent and most successful machines used in
America
is here appended. This machine was
used by the Hon. Thos»
H
d
d y
y
The
jjans should be
2
feet wide, 4
inches
deep, and 8
inches at the Reports
to the Foreign
Office. 167 back
; they
are
laid on
1 x 4 boards, previously
nailed
to runners
; the
height
of the pans above the ground
varies with the height
of the crop
over which the " hopper dozer "
^^ill be taken. The pan should be partly filled with paraffin and water and taken across
the infected crops until full, when fresh oil and water must be added. These machines can be made of any size. These machines can be made of any size. B.
3. MiNEKAL Poisons. Arsenical poisons can be employed to advantage where animals are not
likely to touch them. In America poisoned bran
is successfully employed. Mr. Coquillett
(U.S. Dept. Agriculture) has found
the
folloAving formula
the best
:
1
lb. arsenic,
1
lb. sugar, G
lbs. bran. Add water
to make au
ordinary mash. This
is prepared
as follows
: Mix the dry bran and arsenic
in a tub,
dissolve the sugar
in warm water, and
mix with the
arsenic and
bran. Place this mixture about in
little heaps
;
its action
is not rapid, but always
fatal in about twenty-four hours. (Destruction by Fungoid Disease.) A fungus known as Empvsa
(jrylll found on grasshoppers and
locusts
has been used as a remedy with more or less success. Its introduction into
Egypt might probably be very beneficial, and certainly should be tried. gyp
It
has been imported
into Americs- from
Natal, and
has destroyed
injurious swanns of locusts in Colorado and Mississippi. Dr. Lounsbury (Cape of Good Hope Kept., 189G) refers to this disease
and
its employment, and
says
it causes destruction
to
the swarms when
proper conditions of moisture are present. It has been introduced
into Australia and has met with some success
there also. It has been introduced
into Australia and has met with some success
there also. The method
of employment adopted
by Mr. Froggatt, Government
Entomologist to New South Wales,
is here appended. The method
of employment adopted
by Mr. Froggatt, Government
Entomologist to New South Wales,
is here appended. The method
of employment adopted
by Mr. Froggatt, Government
Entomologist to New South Wales,
is here appended. The fungus must be cultivated in a laboratory on gelatine and sent out The method
of employment adopted
by Mr. Froggatt, Government
Entomologist to New South Wales,
is here appended. The fungus must be cultivated in a laboratory on gelatine and sent out
to operators in test tubes. The operator proceeds
as
follows
:—The fungus should be sent out in
definite quantities, enough
of the culture to make a tumbler full of liquid
being a useful proportion. The
operator should
boil
sufficient water and
let it cool down to luke-warm. The contents of the tube are then extracted
and mashed up with two teaspoonfuls
of sugar and
well
stirred up in the
water with several
bits
of
cork, which have been previously placed in the
glass
as
indicators. Cover the tumbler with a sheet
of paper and then
place it in a warm room and leave for twenty-four hours. AVhen examined,
if
fit for use, the cork indicators should show mycelium growing on them. This culture
is taken
to the
infested
land. Then proceed to catch some
locusts by means of a
net. B.
4. The Afeican Locust Fungus {Empusa fjrijlU). (Destruction by Fungoid Disease.) (Destruction by Fungoid Disease.) The culture placed in a tin is spread over the
locusts and they are
released, when they carry infection to
others and
so
destroy myriads of the pests, The operator proceeds
as
follows
:—The fungus should be sent out in
definite quantities, enough
of the culture to make a tumbler full of liquid
being a useful proportion. The
operator should
boil
sufficient water and
let it cool down to luke-warm. The contents of the tube are then extracted
and mashed up with two teaspoonfuls
of sugar and
well
stirred up in the
water with several
bits
of
cork, which have been previously placed in the
glass
as
indicators. Cover the tumbler with a sheet
of paper and then
place it in a warm room and leave for twenty-four hours. AVhen examined,
if
fit for use, the cork indicators should show mycelium growing on them. This culture
is taken
to the
infested
land. Then proceed to catch some
locusts by means of a
net. The culture placed in a tin is spread over the
locusts and they are
released, when they carry infection to
others and
so
destroy myriads of the pests, Mr. Froggatt, Government Entomologist
of New South Wales,
states
that one tumbler full of liquid is sufficient for 1000 locusts. First Report on Economic Zoology. 1 68 The most favourable
time
to
treat them
is
in the evening—damp
weather if possible being chosen—as the increase of the fungus is doubtfid
unless the air is moist. Further
experiments
should
be
conducted,
however,
before
this
is
definitely considered satisfactory. B.
5. Plants Poisonous to Locusts. 1. Common Garden Larkspur {Delpliinium). 2. Castor Oil Plant {Ricinus communis). These might be employed around
gardens, orchards, etc., as a barrier
to the advance of locust armies. It should be pointed out, however, that stock will eat Larkspur and are
thereby poisoned {vide Dr. E. V. Wilcox's Rept., Bull. 15, Montana Exper. Station, 1897, on " Larkspur Poisoning of Sheep "). B.
G. Natukal Enemies. Locusts
suffer from many natural enemies, both vertebrate and inver-
tebrate. Amongst
the
former may
be
mentioned
fowls
and
turkeys. Droves of the
latter
clear
off locusts very rapidly, as many as
fifty being
found at once
in a turkey's
crop. Encouragement of these birds should
be given in all districts where
locusts abound. Numerous ^vild birds also
feed off locusts. Amongst
insect
enemies
are
numerous
diptera
or
flies,
especially
Tachina
Flies
{Tac/iinidce), and Flesh
Flies
(Sarcophagid(B), Avhose
larvae
or maggots live inside and destroy the young locusts. Many carnivorous flies, such
as the Asilidte, or " Wolf
Flies," feed
off
the young " hoppers." Predacious beetles and
their
larvae devour locusts
in different parts of
the world, especially the locusts' eggs. In North America a
species
of
mite, /. locvstarnm, Riley, is the most
effective enemy of the various locusts. These mites feed
oft" the eggs and
also the winged adults. No natural enemies are able to cope with
locusts, however, unless
it be
the Locust FungU!^ {Eminisa gnjllii) (vide B. 4), (Signed) (Signed) Feed. Y. Theobald. Reports
to the Colonial Office. 169 My first letter on
locusts, dated 28tli August (copy enclosed) addressed
to you, must have been
lost in the
post, as
it
^\as
carefully sent to your
Mdress. I note that
a
separate report
re White Ants
will be forwarded to me
later. I have received a copy of the " Bulletin " for July, 189(>, of the
Botanical Department
of Trinidad throiigh
the
Colonial
Office
; but the
" Bulletin " deals with
the West Indian White Ants
only, which present
no resemblance whatever to the Sudan White Ants. I am anxiously awaiting the result
of
the
investigations kindly under-
taken by the Museum ])efore taking any further steps. I remain, etc.,
(Signed)
Gleichen. I remain, etc.,
(Signed)
Gleichen. B.—REPORTS
TO
THE
COLONIAL
OFFICE.
1. THE MARINE EESOUECES OF THE WEST INDIES. Colonial Office,
Downing Street, S.W.,
IWi AiJiil, 1901. To Pkofessor E. Ray Lankester, LL.D., F.R.S. Sir,—I am directed by Mr. Secretary Cham])erlain
to
transmit
to
you
the
occompanying
copy
of
a
letter
which
he
has
received
from
Dr. Morris, the Imperial Commissioner of xlgriculture for the West Indies,
relating
to
a
paper,
of
which
a copy
is
also
enclosed, on
the Marine
Resources
of
the West
Indies, by
Dr. J. E. Duerden,
Curator
of
the
Museum of the Institute of Jamaica, together with the copy of a despatch
on the same subject from the Governor of Jamaica. 2. Mr. Chamberlain would be glad if you would be so good as
to take
these papers
into your consideration and favour him with your opinions
upon the subject. p
j
3. In accordance with Dr. Morris's request, copies of these papers have
also been referred to Professor Howes. I am, etc, I am, etc,
(Signed)
H. Bertram Cox. (Signed)
H. Bertram Cox. Imperial Department op Agriculture for
THE West Indies,
Barbados,
Uth ll
h
1901 Imperial Department op Agriculture for
THE West Indies,
Barbados,
Uth ll
h
190 Uth llarch, 1901. Sudan Government,
Civil Secretary's OflBce, Cairo,
Zrd November, 1901. Sudan Government,
Civil Secretary's OflBce, Cairo,
Zrd November, 1901. To C. E. Fagan, Esq., Assistant Secretary,
Natural History Department, British Museum Dear Sir,—I beg to thank you for your letter of the 17th October
enclosing some very valuable suggestions for the destruction
of
locusts. They should eventually prove of
the greatest use, though as yet the Sudan
is too new and too thinly populated
to permit
of operations being carried
out very effectively. Reports
to the Colonial Office. Uth llarch, 1901.
To The Under Secretary of State,
Colonial Office.
Sir,—I have the honour to forward, herewith, a copy of a paper on
" The Marine Resources
of the West
Indies." read
before
the
late West
Indian
Agricultural
Conference by Dr.
J.
E,
Duerden,
Curator
of
the
Museum of the Institute of Jamaica. To The Under Secretary of State,
Colonial Office. Sir,—I have the honour to forward, herewith, a copy of a paper on
" The Marine Resources
of the West
Indies." read
before
the
late West
Indian
Agricultural
Conference by Dr. J. E,
Duerden,
Curator
of
the
Museum of the Institute of Jamaica. First Report on Economic Zoology. lyo i. Owing:
to the special interest attached
to the subject, tlie paper has
been
issued
as an extra number of the " West Indian
Bulletin."
It
will
iilso appear amongst the Conference
i)apers
to be published
in
the second
volume of the Bulletin now in the press. 3. Althou2:h
the
British West Indian
Islands
are surrounded by wide
seas,
inhaljited by
large numbers
of edible
fish
of
excellent
quality,
the
methods employed in capture are somewhat i^riniitive, and in no instance is
advantage taken of modern improvements. At present a considerable trade
in salt fish is carried on between
these islands and
British North America,
the annual value of which
is estimated at £234,000. 4. Dr. Duerden, so
far
as
I am
aware,
is
the
first
to draw attention
from
the
scientific
point
of
view
to
the
potentialities
of
the
marine
resources
of these islands. It would, in my opinion, be most valuable
if
the subject could be taken up as a part
of the research work entrusted to
this Department. This would be
in harmony with what has been done
with considerable advantage
at Cape Colony and
in connection with the
recently created Board of Agriculture in Ireland. I estimate that the cost
of adding a Fishery Branch
to this Department would be about £800
to
£1000 per annum. .5. I commend
for
special consideration
the resume given
at the close
of Dr. Duerden's paper (pp. 18 and ID). He rightly points out that the
West Indian Fisheries and the men associated with them have been wholly
neglected by the agencies devoted
to
the improvement and
extension of
the
industrial
resources
of
these
Islands," and he
concludes
as
follows
:
"The
directions
along
which
development and
investigation
in
fishery
matters are most needed
at
present within the West Indies may finally be
summarised
:—(1) The best methods of capturing and curing tropical fish
;
(2) Knowledge of the life-history and
habits
of the edible and migratory
fish
;
(3) Encouragement
of
enterprise
in
fisheries
generally
;
(4) The
best means of shipping
live turtle. Repoi'ts
to the Colonial
Office. Repoi'ts
to the Colonial
Office. 171 King's House, Jamaica,
%th March, 1901,. King's House, Jamaica,
%th March, 1901,. To The Right Hon. J. Chamberlain, M.P., etc. Sir,—I have the honour to transmit
to you a copy of a pamphlet. by Dr. J. E. Uuerden on the subject of
Fisheries
in the West Indies and
to commend
it to your consideration. 2. I
undei-stand from
Dr. Duerden
that
Dr. Morris
has
expressed! himself very fiu-ourably with regard
to the suggestions contained
in the
paper, and
that
it
is
possible he may recommend
that
experiments for
giving
eifect
to them should be carried ouo in connection with, and under
the
auspices
of,
the
Imperial Department
of
Agricultm-e
for
the West
Indies. 3. Were the financial circumstances of Jamaica different
to what they
are,
I should consider
it desirable
that
efforts
should
be made
by the
Colonial Government to improve and develop the fisheries
of
the
island. Any such efforts are, however, for the present, at
all events, " beyond
the
range of practical politics." 4. I
feel, however, that
it would be of great benefit
to the Colony if
experiments in the directions suggested by Dr. Duerden, particularly with
regard
to the
artificial breeding and
culture
of
turtles, could be
carried
out, and I shall be very glad to learn that you have found yourself able to-
sanction any recommendation which Dr. Morris may make
for the matter-
being taken in hand by the Imperial Department of Agriculture.. I have,
etc., (Signed)
Augustus W. L. Hemming,
Governor.. (Signed)
Augustus W. L. Hemming,
Governor.. g,
Governor.. British Museum (Nat. Hist.),
Cromwell Road, S.W.,
3rd May, 1901.. British Museum (Nat. Hist.),
Cromwell Road, S.W.,
3rd May, 1901.. To The Under Secretary of State,
Colonial Office. Artificial hatching and rearing of the
green turtle and
the hawksbill
;
(5)
Restocking of the exhausted grounds
around Barbados with artificially reared
sea-eggs
;
(G)
Oyster, sponge, and
lobster culture. One of the great endeavours of to-day in the West Indies
is to supplement
in
as many
directions
as possible the old
industries
of
sugar and rum by the introduction and encouragement
of other products
;
and in the undeveloped resources of
the sea the Colonies have a possession
which,
if rightly
used,
will
constitute a valuable adjunct
to
the many
agricultural efforts." ('). In order that the subject may be placed before the Secretary of State
in a complete form I suggest that a copy of this letter and of Mr. Duerden's
paper be referred for
their opinion as Zoological Experts to Professor Ray
Lankcster, F.R.S., Director of the British Museum (Nat. Hist.), and to Pro-
fessor Howes, F.R.S., of the Royal College of Science, South Kensington. 7. In the meantime copies of Dr. Duerden's paper have been communi-
cated
to the Governors and
to
all
the leading
officials and
residents
in
these Colonies. 7. In the meantime copies of Dr. Duerden's paper have been communi-
cated
to the Governors and
to
all
the leading
officials and
residents
in
these Colonies. 8. I forward, under separate cover, five extra copies of the Report for
the use of the Colonial office. 8. I forward, under separate cover, five extra copies of the Report for
the use of the Colonial office. I have, etc. I have, etc. have, etc. D. Morris,
Commissioner of Agriculture for the West Indies, I have, etc. (Signed)
D. Morris,
Commissioner of Agriculture for the West Indies, (Signed) o
s,
Commissioner of Agriculture for the West Indies, Abstract of Dr. Duerden's Report on the Marine
Resources of the West
Indies. In an extra number of the West Indian
Bulletin
issued
in
1001, Dr. J. E. Duerden reviews
the
chief
marine
resources
of
the
British West
Indies. In
this he gives an account
of the fisheries of the West Indies,
Und points out that in a few instances only are the marine products of any
export value to the Colonies, whilst on the other hand there
is an enormous
import trade in dried and preserved fish. The principal marine
resources
are as folloAvs
:
Turtles, Jamaica being
the
chief centre
of
the West
Indian
turtle
trade,
the exports
for lOOO
being about £10,000. The industry is concerned with the two well-known
species,
the
Green
Turtle
{Clielone
mkias) and
the
Hawksbill
{Chelone
imbrkata). The
supply
is
chiefly
ol)tained
from
around
the Cays and
Mosquito coast
of Central America. The Report shows
that there
is an
evident diminution
in
the
supply, merchants never being
able
to obtain
sufficient
to
meet
the
export demands. The two
subjects dwelt upon
concerning turtles of great importance are
their
artificial rearing so as to
produce them in greater numbers to meet the demand and the best method
of shipping them. The mortality from capture to
landing
in England
varies from as much as 2b to oO per cent. Under the
heading
of " fishing "
is
given
a general account of the
industry, and
stress
is
laid
on
the
primitive methods adopted by the
fishermen
:
" fishing as adopted
elsewhere
is comparatively neglected and
undeveloped."
The amount
of
coral
prevents
trawling, and moreover
there
are evidently not enough
flat
fish
to make
this method pay. The
use
of seine and
other
nets along the shore and shallow banks
is very
profitable
; but unfortunately the
habits of the schools of migratory fish,
such
as June fish,
herring, sea
mullet, etc., are not known, nor the
best
means of catching them, nor of preserving them when caught. The fishing
industry appears to be mainly in the hands of natives. Amongst the chief
fish
of good
quality
are various
species
of Snappers {Mesoprion), Yellow
Tail {Ocyunis chrysurus), Grunts (Rmmdo/i), Silks {J'ropidiurus dentatus),
King and June fish, river and marine mullet and CaUperus {MugiT), and
Snook iCentropomus). They occur around
Jamaica, where
the average
price of fish
is
^^d. a pound. Barbados
is celebrated for
its large flying-
fish industry. British Museum (Nat. Hist.),
Cromwell Road, S.W 3rd May, 1901.. To H. Bertram Cox, Esq.,
Colonial Office, Downing St., S.W. To H. Bertram Cox, Esq.,
Colonial Office, Downing St., S.W. Sir,—In
accordance
with
Mr. Secretary
Chamberlain's
request,
conveyed
in your
letter
of
April
29th,
I have
considered
the
letter
of
Dr. Morris and
Dr. Duerden's Report on
the Marine
Resources
of
the^
West Indies, of which you were good enough to send me copies. The matter referred to has been for some time under my consideration^
and I find myself
in entire agreement with Dr. Morris. I think it would
be a most valuable step in
the
public interest were the Marine Resources
of
the West
Indian
Islands taken up
as
a part
of
the
research work
entrusted to the Imperial Department of Agriculture for the West Indies. p
The paper by
Dr. Duerden on " The Marine
Resources of
the West
Indies "
is a valuable
one. The various sources of wealth in
the seas of
the West
Indies
are each
carefully sketched. By the references made to
marine
investigations carried on elsewhere, Dr. Duerden shows that he
is
well informed
in
the subject, whilst his observations and suggestions and
general handling
of
the
subject show
originality and
full
competence. Dr. Duerden has had a thoroughly sound
training as a scientific biologists
His original papers, as well as the present report on the Marine Resources-
of the West
Indies, prove him
to
be
a
trustworthy scientific adviser, who First Report on Economic Zoology. 172 would
be regarded with respect and
confidence
l)y
scientific men
in
this
"country were he further employed in connection with this subject. I am
of
the
opinion
that
the
carryini«:
out
of
the
investigations
Suggested by
Dr. Duerden's
Report, under the
auspices
of
the
Imperial
iDepartmeht of Agriculture for
the West Indies, would
lead to economic
l-esults
of
practical value and
justify
the expenditure of public funds in
that direction. I have, etc.,
(Signed)
E. Rav Lankester. I have, etc., (Signed)
E. Rav Lankester. Reports to the Colonial
Office, I'^y Under the heading
Oysters we
find
that
the West
Indies
have
an.,
oyster
in abundance, Ostrea parasitica, Gmelin, which grows on the roots
of mangrove trees. There
is no system
of cultivation at present. They
are much appreciated and
find a ready sale. Many other edible mollusca
occur, including the Mussel
(M>/tiliis
exnstus, Linn., the
Scallop {Pecten
zigzag, Chemn.), various " ark " shells. Area- spp. ? and C'odahia tigerina. Lobsters, shrimps and
crahs are also amongst the marine resources, the
most abundant
lobster
being PaUntirus
arg^s,
Latr. They and other
species are caught
in fish-pots from amongst the coral. It
is pointed out
that successful cultivation
of
lobsters
might
be
carried
out
just
as
in
Newfoundland and Canada. Sea-eggs. Barbados occupies an almost unique position
in having an
important industry founded on the marine forms of
life called sea-urchins,
or sea-eggs. Its annual value
is
estimated' at nearly £4,000. The roe or-
reproductive organs, are the
part used as food. There
is unfortunately a
great decrease in the number of these Echinoderms. The chief species is. known as
Hijyponoe
csculenta, Leske. They form a
stapJe food for a few
months along the coast. Before any remedial measures can be adopted,
it
is necessary to know the life-history of this sea-egg. Hohthurians,
Beche-de-Mer,
Trepaiig,
etc. (It'eat
numl)ers
of
these
echinoderms occur on the
floors
of
the
seas
in. the W'est
Indies. The
species have not been
identified, but the Jamaican ones are of the genera
Holothnria and SticopJtus. Experimental shipments of Be'che-de-Mer were
carried
out a few
years
ago
at
the
Caico? Islands
with
the
object
of
supplying the American Chinese with
their favourite
article of diet. To
show the importance
of
this marine animal, the report mentions that the
annual export value to Queensland
is about £23^000. Abstract of Dr. Duerden's Report on the Marine
Resources of the West
Indies. The Flying-Fish
{Exococtus
roberti)
is estimated
to
yield
annually £13,000. Snappers and Brines {Centropristes
ocidatus) are
also
taken in numbers by line fishing around Barbados. Reports to the Colonial
Office, First Report on Economic Zoology. First Report on Economic Zoology. 174 The resume is given iu Dr. Morris's letter, p. 188. The report has three
appendices
: pp
(1) The Fisheries of Barbados, where we learn the approximate income
is £ll>,50(i. (2) The Jamaican Fisheries
; an account of the operations in Jamaica
of
the Caribbean Sea
Fisheries Development
Syndicate,
Limited, which
does not appear to have been financially successful. (3) A Report on the Sea-egi,' Industry of Barbados. (3) A Report on the Sea-egi,' Industry of Barbados. 2. CEYLON PEAEL FISHEKIES. Colonial Office,
Downing Street,
Ith August, 1900. To The Director of the British Museum (Nat. Hist.). Sir,— I am
directed by Mr. Secretary Chamberlain
to
enclose for
your consideration copies of a special Report on the Ceylon Pearl Fisheries
and of the
last report on the
inspection of the
pearl
banks, which have
been received from the Governor of that Colony. Mr. Chamberlain would be much obliged
if you would be good enough
to advise him on the subject generally, but
I am
to
state that he doubts
whether
the
Colonial
(lovernment would
be
well
advised
to
incur any
considerable expense in the matter, unless
it
is
considered
to be of
great
scientific
interest,
as
the
local
conditions seem
to
be
well
known
for
practical purposes. The Governor of Ceylon has suggested
that the opinions of the Royal
Society, British
Association, and
Zoological
Society should be
invited
as
to
wliether
it
is
desirable
that
these
fisheries should
be
inspected
by a
scientific expert, and also
that Dr. Herdman, F.R.S., should be consulted,
but no application has yet been made to these Societies or to Dr. Herdman,
pending an expression of your opinion. I am to add
that various
prints
relating
to
Pearl
Fisheries in Ceylon
can be seen in the Library of this Office. I am, etc.,
(Signed)
C. P. Lucas. British Museum (Nat. Hist.),
Cromwell Road, S.W.,
\Wi August, 1900. SpON&ps, The West
Indies and
Florida, along with the Mediterranean, are the
principal sponge producing areas of the vforld ;.but fine bath sponges also,
come from Australia. The shores around the Bahamas
ai-ie the best known sponge grounds in
the
West
Indies. They
form
the
greatest
industry
of
that
Colony. Dredging and diving for them have been prohibited. They are gathered
by means of two-pronged forks attached to staves 2.5 feet
in length. The
sponge exchange
is
at Nassau. The annual value
is
nearly £100,000. The United States Government has undertaken the
investigation
of the
Florida sponge grounds with
a
view
to the
l)etter development
of
the
industry. Successful experiments in
, the
artificial propagation of sponges
by transplanting and by cuttings have been carried out in the Mediterranean
and in Florida. Amongst other
industries mentione.d
in the report
are
the whale
oil
industry, still carried out
in a small degree around isome of the islands of
the Lesser Antilles. Ambergris
is occasionally found on the coasts of the
Bahama Islands. Companies
of
dolphins
are
often
sepn, traversing
the
length
of
Kingston Harbour, but no attempt
is ever made
to secure them. The
Manatee
is also sometimes caught and
the
flesh
sold,^ byt they breed too,
slowly to become of n^uch ep.onomic, importance. British Museum (Nat. Hist.),
Cromwell Road, S.W.,
\Wi August, 1900. To The Right Hon. Joseph Chamberlain. Sir,—I have read the Special Report on the Ceylon Pearl Fisheries
forwarded
to me by Mr. Lucas at your request. I am of opinion that the
recommendations made by
Sir W. Twynam are well based and should, so
far as I am able to judge, be carried into effect. The questions connected
Avitli
the proper management and
fishing of
pearl
oyster
banks and
other
similar submarine
som'ces
of
wealth
are
of
great
scientific
interest, and
should,
in my
opinion, be
continually
investigated and
dealt with
in
the
interest of
the community. Results Repoi'ts
to the Colonial
Office. 175 obtained in Ceylon may be found
to be of value from a commercial point
of view in Queensland or again in the West Indies (Sponge fisheries). I am
decidedly
of
the
opinion
that
the
expenditure
of
a
certain
proportion
of
the
revenue
derived
from
the
Ceylon
Pearl
Fisheries,
upon thorough and
authoritative study of
the questions raised
in
Sir
AV. Twynam's report by
the
best scientific
naturalists whose
services can
be
obtained
must
in
the
course
of time—if
persisted
in
and
sufficiently
supported by mouey needed
for experiments and
investigation—produce
a valuable return
to
the
State
in
the form of
increase
in commercial
results. A brief inspection of the banks by a capable scientific naturalist or the
employment
of
a
second-rate
man
of
no
real
scientific
knowledge
or
training would, in my judgment, be a waste of public money. I should myself like to see Professor Herdman, of Liverpool, entrusted
with
a two
or
three
years' mission
in
connection with the Ceylon
Pearl
Fisheries. He
has given
special
attention to oysters and
oyster fisheries,
and
is
a man
of
genuine
knowledge and
also
possessed
of
energy and
initiative. It would be
possible for him to give
three or four montlis
in
each year to his professional work in England and to spend
the rest of
the
year (at the ])roper season) in Ceylon. I think that Sir W. Twynam's report might very well be submitted
to
Professor Herdman for his opinion, and
that before taking a definite step
it might be
well
to submit
his proposals
to
the
Council
of
the
Royal
Society for
their
advice. But
I
should
not recommend
that
either
the
Zoological Society or the British Association be consulted. It
is evident from
Sir W. British Museum (Nat. Hist.),
Cromwell Road, S.W.,
\Wi August, 1900. Twynam's report
there
are many matters
connected with
the pearl banks upon which a competent naturalist versed
in
marine
biology
could
at
once
clear
up
doubt. For
instance,
the
mysterious enemy of the
oysters mentioned
in the
report, which
drills
a
small round
hole
in
their shells. Every marine
biologist knows
at once
that
this must be one of the whelk-like gastropods, which preys upon
the
pearl
oyster as do
its congeners
in European
seas
prey upon European
oysters and comb-shells. Were
a competent
naturalist,
such
as
Professor Herdman, entrusted
with
a thorough study of
the Ceylon
pearl
banks, and
provided
with a
well-fitted steam cruiser for dredging, sounding, diving, etc., there can be
no doubt that, in the first place, zoological results of great general
interest
would be obtained, as well as collections of value to the national Museum,
and new facts of
the most varied kind tending to advance our knowledge
of marine life. I
believe, moreover, that
in
the second
place such knowledge of the
facts would be definitely gained
as would enable
the Ceylon Government
to improve the pearl fisheries and to manage them in the best possible way
with a view to getting the proper commercial return from them. It
is
impossible
to
foretell
what
results
a
clever
naturalist might
obtain. The
artificial reanng
of
the
spat
of
the
jiearl
oyster and
the
nursing and transference of the young oysters as carried out in regard
to
European
oysters might
be found
possible and
of immense commercial
value. Finally
the
artificial
production
of
pearls
is
always,
as
far
as
zoological
science
enables us
to form an
opinion, a
possibility. Perhaps
I may, in conclusion, be allowed
to point out
that, some thirty-five years First Report on Economic Zoology. 176 ago, Jiu experimental inquiry
into the ]iearl fisheries of Ceylon, which was
initiated
l)y (iovernment, ended
in
failure and disappointment, owing
to
the
fact
that
the
matter
was
entrusted
to a gentleman
who, though
acquainted with
sea-fishing
as a sportsman, had no
scientific knowledge
or training. g
During the past
thirty-five years our knowledge of the treatment of
oysters and similar questions has vastly increased. If a
naturalist who
is
really worthy of
trust and conversant Avitli the
snbject
is sent
to Ceylon
to
study
the
pearl hanks,
it
is, in my
opinion,
highly probable that the expenditure involved will
l^e amply repaid
l)y the
results. CoLONiAri Office,
Downing Street, CoLONiAri Office,
Downing Street,
23rrf August, 1900. 23rrf August, 1900. To The Director, British Museum (Nat. Hist.). To The Director, British Museum (Nat. Hist.). Sir,—I am directed by Mr. Secretary Chamberlain to acknowledge
the receipt of your letter of
the
l;-»th instant and to thank you for your
advice on the subject of the Pearl Fisheries of Ceylon. 2. Mr. Chamberlain
will communicate with Dr. Herdman and subse-
quently with the Royal Society, as you suggest. 3. I am to ask
that you
will be
so good
as
to
retiu'n the Reports
enclosed
in the
letter from
this Department of the 7th instant, as there
are no
other
copies
of these
prints available. The Governor of Ceylon
has been asked
to send further copies, which will be forwarded
to you as
soon as they are received. I am, etc.,
(Signed)
C. P. Lucas. I am, etc., (Signed)
C. P. Lucas. British Museum (Nat. Hist.),
Cromwell Road, S.W.,
\Wi August, 1900. Such a man could not be obtained
for a
less payment than one
thousand
pounds
a
year,
exclusive
of
all
expenses
;
and
it
would
be
necessary to employ him for three years at least. I am, etc., I a , etc.,
(Signed)
E. Ray Lankester. (Signed)
E. Ray Lankester. Abstract of Report on the Ceylon Pearl Fisheries.
By Sir W.
C
Twynam, K.C.M.G.
(Colombo,
189'.)). By Sir W. C. Twynam, K.C.M.G. (Colombo,
189'.)). In this long report
of
sixty-six
pages, Sir W. C. Twynam
first
points
out the injurious nature of currents and foul water to the pearl oyster. For some years the real spat of the pearl oyster does not seem to have
been known,
the
spat
of
other
Avicuhe being taken
for
young
pearl
oysters. y
The enemies
of the
oyster are given, amongst
them the following
:
shellfish,
the
chank
of
commerce
{Turbinella
pi/non),
the
horse
and
elephant chanks {PyrvJa carnaria and Murex regius). A small mussel {Modiola) known as the Suran spreads a kind of blanket
over the oysters and suffocates them
;
this
is, however, rare in the Ceylon, Reports
to the Colonial
Office. 177 beds. The crab
is
also
said
to be
injurious,
cutting
the byssus
of the
oyster. A
note
is given on page
6
regarding an enemy
that makes
a
round
hole
in
the
oyster
shell
;
this mysterious enemy
is
one
of
the
carnivorous whelks. Evidently, from the report, numerous small mollusca prey on the pearl
oysters. Two
fish, the Trigger Fish {Balistes
mitis) and
skates (Trijgoii
irarnah), also do much harm. On page 5
it
is stated that " the numerous
rock fish which abound on the Arippu banks feed on oysters
. . . the quan-
tity devoured by these voracious fish must be considerable."
Later,
it is
stated to be useful, as it preys on the injurious Suran or mussel. Skates of
several unknown species are referred to as very destructive. Divers, both
European and
native,
give various
tales
as
to the damage done by
sea
snakes, but nothing authentic is given. Floods of fresh, muddy water are stated to be most injurious. Little definite seems
to be recorded as to the age of pearl
oysters, but
it
is stated " that
oysters may be profitably fished at the age of four years,
and
that they are in
their prime at five years, and may be kept
till that
age if circumstances permit of
it, but if they are kept until the sixth yea-
they are almost
certain to
be found
dead."
The best time to fish then:
however, does not appear to be settled. The
advisability
of retaining
native
divers
is entered
into
at
some
length,
their
superiority
over
the European
at
this work
being
clearly
pointed
out. Abstract of Report on the Ceylon Pearl Fisheries.
By Sir W.
C
Twynam, K.C.M.G.
(Colombo,
189'.)). Their reward
is now
raised
to
one-third
of
the
oysters
collected. Recommendations to
start a chank
fishery in
the neighbour-
hood of the pearl banks are
given. One
fishery exists north of Manaar
Islands, about 2,000,000 chanks being exported from Jaffna
to Calcutta. Ths chanks are used as ornaments by the Hindus. The main body of the report (39 pages) is taken up by eight appendices. The first dealing with spat, true and
false
;
enemies of
the oyster
; chank
fishery
; age of the pearl oyster and artificial culture
;
being extracts from
the
report
of Mr. Thomas, Madras
Civil
Service, to
the Government
of
Madras, on the Pearl Banks and Fisheries of Tuticorin. The most important
part in this report regarding
the true spat
is here
reproduced
: " The
challenged
spat
in
the
largest
shell
which
I
have
seen
is
4-i
sixteenths
of
an
inch
from
binge
to
contour
rectangularly
at
its
widest
point, and
the
largest
drawing
in
Sir
J. Emerson
Tennent's
work
is
no
more
;
it
is,
therefore,
so
small
as
to
need
very
close
examination. Looked
at
under a hand
lens
and under
a low power
microscope,
I
made
it
out
to
differ from
the
shell
of the pearl
oyster
in being much more
convex, more
oblique
;
in having
the
ear on
the
short
side,
not produced
in an
almost
straight
line,
but rounded
off
and turned up
instead
of
being
flat
;
in having the right
valve
fitting
deeply into the left
valve, with the edge of the right valve turned back at
about an angle of 45° for the whole contour in some, for others only from
the
sinal
ear to
half way round
the contour,
instead
of
_ the two
vaJre^i
meeting each other nearly
fiat, as in the pearl oyster
,
in having none of the
spines with which the
pearl
oyster
is
covered, and
distinctly
different
flanges
;
in having no algce adhering to
it
;
in having the umbones more
anterior or advanced beyond the hinge line
;
in adhering to weed, said
to
be Saragossum vidgare, instead
of to rock and such-like hard substances First Report on Economic Zoology. 178 in being
differently coloured, the dark
lines of colour radiating
as in the
drawing from the convexity to the contour. * The
figures
given
in
Tennent's
' Natural
History
of
Cevlon
'
are
therefore
wrong.—F.V.T. Abstract of Report on the Ceylon Pearl Fisheries.
By Sir W.
C
Twynam, K.C.M.G.
(Colombo,
189'.)). Again,
it is only the concave
part of the shell that
is coated with glistening
nacre, the broad deflected
margin being dull. It
is not
so
in the
pearl
oyster, in which the nacre
comes
close
to
the
margin. In
the
aviculce, of which our
pearl
oyster
Avicula {Meleagrina) margaritifera is one, the prolonged hinge line, straight
at the hinge,
is brought in below with a curve that gives it a similitude to
the wing of a bird, and the sinal ear, though shorter, is also slightly curved
in below. In some
avicidce. this
formation
is
more
expressed
than
in
others, so that they are divided
into two sections of the long-Avinged and
short-winged
avkulce. Avicula macroptera
is the type of the former, and
Avicula
heteroptera and
A. crocea
are
illustrations
of
it
;
of
the
latter
A. margaritifera is the type, but still has the peculiarity distinctly present. In the challenged spat
it
is wholly absent. At the same
time, however,
that
it
is
said
in
Reeve's
" Conchologia
Iconica "
that
this
feature
is
always
present
in
the
aviculce,
it
is
not
shown
in
the small
shell
of
A. vexillum, figured magnified
in
this work, which, as far as the drawing
goes, has a general similitude to the challenged spat and has against
it the
remark " Habitat, Ceylon (in deep water), Gardner," but beyond
this
the
text
description, though very
brief, hardly
tallies, and
there
are
to mv
thinking three,
if not four, forms among the challenged spat, sll of whict
show under
the
microscope "the
prismatic
cellular
structure
of
shell
found in most of
the aviculm " (Carpenter). My belief
is that
they have
been so long sailing under the false colours of being
the pearl oyster spat,
that
they
are unnamed
and
seemingly mature
aviculce,
but
I am
not
concerned to name them
; all my contention for the purposes of this report
is that they are not pearl oysters.* This is pointed out
as having an important bearing on the
suppose<l
disappearance of young pearl oysters from certain beds. * The
figures
given
in
Tennent's
' Natural
History
of
Cevlon
'
are
therefore
wrong.—F.V.T. 179 EPUII'l'lGEUlD.E. Uromenus,
Bol. S. France,
Corsica,
Sardinia, Algeria. rugosicollis, Serv. S. France, Cor-
sica, Sardinia, Algeria. costaticollis, Luc. Algeria. laticolUs, Luc. Algeria. Finoti, Br. Algeria. agarena, Bol. Ceuta. latipennis, Fisch. Algeria. Uromenus,
Bol. S. France,
Corsica,
Sardinia, Algeria. rugosicollis, Serv. S. France, Cor-
sica, Sardinia, Algeria. costaticollis, Luc. Algeria. laticolUs, Luc. Algeria. Finoti, Br. Algeria. agarena, Bol. Ceuta. latipennis, Fisch. Algeria. Uromenus,
Bol. S. France,
Corsica,
Sardinia, Algeria. PHASGONUEID^. STENOPELMATrD.E. Lezina, Walk. concolor, W. Egypt. Magrettia, Brunn. abominata, Bruua. Suakini, Don-
gola. obscura, Burr. Somali. Hetbodid^. Oyminj][>roctus, Karsch. Maurelii, Luc. Senegal, Soudan. abortiva, Serv. S. and W. Africa. Anepisceptus, Fieb. horridus,
Burm. Egypt,
Syria,
Arabia. Servillei, Reiche, Abyssinia, Somali. Revoilii, Luc. Somali. Hippolyti,
Kb. {Servillei,
Luc). Abyssinia. Buspolii, Schulth. Somali. Suakimensis, Kb. Suakim. Roheckii, Schulth. Somali. Etigaster, Serv. spinulosus, L. Morocco. Woodii, Kb. Somali. Powysi, Kb. Morocco. Ouyoni, Luc. Algeria. STENOPELMATrD.E. Madiga, Kb. aberrans, Schulth. Somali. Magrettia, Brunn. abominata, Bruua. Suakini, Don-
gola. obscura
Burr
Somali Bradyopisthius, Karsch. paradoxurus, Karsch. Bradyopisthius, Karsch. paradoxurus, Karsch. MEC0P0DID.E. Ariagona, Krauss. Margaritx, Kr. Tenerifte. Pachysmopoda, Karsch. ahhreviaia, Tasch. Sokotra. Clenodecticus, Bol. Bolivar i, Targ. Sardinia, Oran. Vasorensis, Fin. N. Africa. Euthyphlebia, Schulth. parallda, Sch. Somali. Pholldoptera, Wesm. pundifions, Bumi. Egypt, Syria. Phanekopterid^. Odontura, Kamb. spinulicauda,
Eanib. S. EurojiC,
Algeria. Borrei, Bol. Algeria. algerica, Brunn. Algeria. quadridentata, Krauss. Algeria. terniensis, Fin. Algeria. Tetl igonia, Linn. ulhifrons, Fabr. Europe, Madeira. GoXOCEPHALID.r.. Platystolus, Bol. pachygaster, Luc. Algeria. Pycnogaster, Graells. Finoti, Bol. Algeria. Conocephalus, Thunb. nitidulus, Scoj). S. Euroj e, Afric a
Algeria, Canaries, etc. Meconemidji:. Becticus, Serv. griseus, L. Europe, Madeira. laticuuda, Brunn. Sicily, Algeria. tessellaia, Charp. Europe,
Algeria. seniai, Fin. N. Africa. KabyJa, Fin. N. Africa. OropTiila, Krar.ss. mihigena, Kr. Tenerift'e. DECTIC1D.E. Xiphidium, Serv. concolor, Burm. Hungary, Egypt. stramineum, De Haan. Egypt. luguhre, Redt. Egypt. Somali, Burr. Somali. conocephalus,
L. N., W. and
IL
Africa, Madagascar. Brymadusa, Stein. faUaciosa, Fin. Algcr'a. Pterolepis, Ramb. Gessardi, Bouu. Tunis. indigena, Fin. Algeria. BhacocJeis, Fieb. maura, Borm. Tunis. Hetbodid^. Oyminj][>roctus, Karsch. Maurelii, Luc. Senegal, Soudan. abortiva, Serv. S. and W. Africa. Anepisceptus, Fieb. horridus,
Burm. Egypt,
Syria,
Arabia. Anepisceptus, Fieb. horridus,
Burm. Egypt,
Syria,
Arabia. Anepisceptus, Fieb. horridus,
Burm. Egypt,
Syria,
Arabia. Servillei, Reiche, Abyssinia, Somali. Revoilii, Luc. Somali. Hippolyti,
Kb. {Servillei,
Luc). Abyssinia. Buspolii, Schulth. Somali. Suakimensis, Kb. Suakim. Roheckii, Schulth. Somali. Etigaster, Serv. spinulosus, L. Morocco. Woodii, Kb. Somali. Powysi, Kb. Morocco. Ouyoni, Luc. Algeria. Servillei, Reiche, Abyssinia, Somali. Ephippigera, Latr. transfuga, Brunn. Algeria. nigromarginata,
Luc. Morocco,
Algeria. compressicolUs, Fiscb. Algeria. antennata, Brunn. Algeria. innocenfii, Fin. Tunis. oudryanus, Bonn. Tunis. vosseleri, Krauss. Algeria. trilineata, De Haan. Tripoli. Vaucherinna, Sauss. Morocco. N
2 Revoilii, Luc. Somali. Hippolyti,
Kb. {Servillei,
Luc). Abyssinia. Suakimensis, Kb. Suakim. Roheckii, Schulth. Somali. Etigaster, Serv. spinulosus, L. Morocco. Woodii, Kb. Somali. Powysi, Kb. Morocco. Ouyoni, Luc. Algeria. Powysi, Kb. Morocco. Powysi, Kb. Morocco. Ouyoni, Luc. Algeria. N
2 Appendix. i8o Calliphona, Krauss. Konig!,, Kr. Canaries. AUuaudi, Kr. Canaries. Mauretanica, Sauss. Morocco. maroccana, Sauss. Morocco. lobata, Sauss. Algeria. tairiiata, Sauss. Morocco. hastata, Sauss. Morocco. Calliphona, Krauss. Konig!,, Kr. Canaries. AUuaudi, Kr. Canaries. Mauretanica, Sauss. Morocco. maroccana, Sauss. Morocco. lobata, Sauss. Algeria. tairiiata, Sauss. Morocco. hastata, Sauss. Morocco. Mauretanica, Sauss. Morocco. maroccana, Sauss. Morocco. lobata, Sauss. Algeria. tairiiata, Sauss. Morocco. hastata, Sauss. Morocco. Sagid^
A^a^a, Charp. ornata, Burm. Egypt. Steropleurus, Bol. scllujer,
Charp. Spain,
rorlugal,
N. Africa (?)
I.ucasi, Briinn. Algeria. Algerica, Bruun. Algeria. PnASGONURlDiE. Phasgonura, Westw. Europe, N. Africa. viridissima, L. N. and W. Africa. Savignyi, Luc. Algeria. margini/era. Walk. Africa. maroccana, Bol. Tangier. algerica, Bol. Algeria. Pseudisotima, Brum. punctata, Br. Somali. Peropyrrhicia, Brunn. Massaice, De Borm. Abyssinia. maculata, Schultli. Somali. Fitmenymus, Pict. Vaucherianvs, Pict. Morocco. Fitmenymus, Pict. Vaucherianvs, Pict. Morocco. Leptophyes, Fieb. Antinorii, De Borm. Sliire. Appendix. .181 Phaneroptcra,
Si.'rv. falcata. Scop. S. Europe, N. Africa,
W. and Central Asia, Madeira. nana,
Cbarp. S. Europe,
E., W. and
S. Africa (can bardly
fail
to
occur in N. Africa). minima, Brunn. Egypt. punctata, Burr. Somali. Epiphlehus, KarscL. crypterius, Karsch. Somali. Buspolii, Scbultli. Somali. Peronura, .Scliultb. Somali, Sell. Somali. Rivx, Sell. Somali. Rhegmatopoda, Brunn. Feeli, Burr. Somali. Acrometopa, Fieb. Servillei,\
Brull^. S. Europe,
Egypt. Conchotopoda, Karscb. Buspolii, Scbiilth. Somali. Eutlujphhhia, Scbultli. parallela, Scb. Somali. MyrnieocopJiana, Bruun. falJax, Br. Soudan. Oonatoxia, Karscb. macuJata, Karscb. Somali. Phaneroptcra,
Si.'rv. falcata. Scop. S. Europe, N. Africa,
W. and Central Asia, Madeira. nana,
Cbarp. S. Europe,
E., W. and
S. Africa (can bardly
fail
to
occur in N. Africa). minima, Brunn. Egypt. punctata, Burr. Somali. Milititsa, Burr. Somaliensis, Burr. Somali. Diogena, Biimn. Fausta, Burm. Xubia, Aden. Tylopsis, Fieb,
lilifolia,
Fabr. Mediterranean
Region, Egypt, etc. perpulchra. Burr. Somali. Karschiana, Schultb. Somali. D b
W lk Peronura, .Scliultb. Somali, Sell. Somali. Rivx, Sell. Somali. Rhegmatopoda, Brunn. Feeli, Burr. Somali. Acrometopa, Fieb. Servillei,\
Brull^. S. Europe,
Egypt. Milititsa, Burr. Somaliensis, Burr. Somali. Diogena, Biimn. Fausta, Burm. Xubia, Aden. Conchotopoda, Karscb. Buspolii, Scbiilth. Somali. Tylopsis, Fieb,
lilifolia,
Fabr. Mediterranean
Region, Egypt, etc. perpulchra. Burr. Somali. Karschiana, Schultb. Somali. Eutlujphhhia, Scbultli. parallela, Scb. Somali. MyrnieocopJiana, Bruun. falJax, Br. Soudan. Oonatoxia, Karscb. macuJata, Karscb. Somali. Oonatoxia, Karscb. macuJata, Karscb. Somali. Debrona, Walk. angiistipennis, Burr. Somali. .. ACRIDUD.E. Acrydiam, Geofif. Nobrei, Bol. Portugal, Morocco (?)
bipunctatum, L. Europe, Algeria. ceperoi, Bol. Cadiz, Tangier. depressa,
Bris. Europe,
Zanzibar
(certainly all N. Africa). annulata, I'bunb. Algeria. variabilis, Klug,
Egypt, Syria. «e&MZosa, Tbunb,
(= unguiculala,
Ramb.). Arabia,
Old
World,
Algeria, Egypt, etc. grandis,
Klug. Egypt,
2\ubia,
Quetta. Paratdtix, Bol. meridionalis,
llamb. S. Europe,
Egypt, Nubia. scalaris,
Klug. Africa, W. Asia,
Egypt, Canaries, etc. Coptottttix, Bol. rufipes, Bol. Somali. Oxycoryphus, Fiscb. compressicornis,
Latr. S. Europe,
W. Asia, Egypt, Algeria, Senegal. venustus, Walk. Cairo. LOCUSTID.E. nasutus, Linn. Morocco (?), Somali. miniiita, Klug. Upper Egypt. annulata, I'bunb. Algeria. variabilis, Klug,
Egypt, Syria. «e&MZosa, Tbunb,
(= unguiculala,
Ramb.). Arabia,
Old
World,
Algeria, Egypt, etc. grandis,
Klug. Egypt,
2\ubia,
Quetta. scalaris,
Klug. Africa, W. Asia,
Egypt, Canaries, etc. nasutus, Linn. Morocco (?), Somali. miniiita, Klug. Upper Egypt. annulata, I'bunb. Algeria. variabilis, Klug,
Egypt, Syria. «e&MZosa, Tbunb,
(= unguiculala,
Ramb.). Arabia,
Old
World,
Algeria, Egypt, etc. grandis,
Klug. Egypt,
2\ubia,
Quetta. scalaris,
Klug. Africa, W. Asia,
Egypt, Canaries, etc. TuYXALIDiE. Tryxalis, Bol. turritus,
L. S. Europe,
Asia,
Africa
(Egypt,
Nubia,
Algeria,
etc.). giganteus,
Fuessly. S. Europe,
Egypt. Uneatus, Tbunb. Morocco. bicolor, Tb. Egypt, Arabia. [N. G. (near last, but b. w. rudi-
mentary).]
tereticornis, BruUe. Canaries. Acrida, Linn. Plmraonis,
Klug. Upper
Egypt,
Bagbdad. ensis, Burr. Sokotra. Duronia, St&l. fracta, Krauss. Egypt. lucasi, Bol. Algeria. laurse, Borm. Tunis. savignyi, Krauss. Egypt. Cldrista, Karscb. flexuosa, Scbultli. Somali. ,
Paracinema, Fiscb. tricolor, Tbunb. S. Europe, Africa,
Algeria. sylvestris, Tbunb. Algeria. Ochrilidia, Stal. tibialis,
Fieb. S. Europe,
Egypt, Paracinema, Fiscb. tricolor, Tbunb. S. Europe, Africa,
Algeria. Tb
b
Al
i sylvestris, Tbunb. Algeria. Ochrilidia, Stal. tibialis,
Fieb. S. Europe,
Egypt,
Syria. Appendix. 1 82 Brachycroiaphus, Krauss. tryxalicera. Fab. Brachycroiaphus, Krauss. tryxalicera. Fab. (Edipoda, Latr. gratiosa,
Serv. S. Europe,
Asia,
Egypt, Canaries. cgerulescens,
Linn. Europe,
W. Asia, Africa, Zanzibar. fuscocinda,
Luc. Sicily, Algeria,
Canaries, Tunis. canariensis, Krauss. Canaries. Mauretania, Luc. Algeria. Arcyptera, Serv. hispanica, Ramb. S. France, Spain,
Algeria. cgerulescens,
Linn. Europe,
W. Asia, Africa, Zanzibar. Stenohothrus, Fisch. pulvinatvA,
Fisch. S. Europe,
Algeria. lxtu&, Walk. Cairo. Bonneti, Bol,
Tunis. amcena, Bris. Algeria. Jjucasi, Bris. Algeria. Simonyi, Krauss. Canaries. fuscocinda,
Luc. Sicily, Algeria,
Canaries, Tunis. Bonneti, Bol,
Tunis. Thalpomena, Sauss. Algeriana, Luc. Algeria. Maderm, Serv. Madeira. Picteti, Krauss. Teneriffe. amcena, Bris. Algeria. Jjucasi, Bris. Algeria. Simonyi, Krauss. Canaries. Picteti, Krauss. Teneriffe. Stauronotus, Fisch. cruciatus, Pall. S. Europe, Morocco,
Egypt, Algeria. Oenei, Ocsk. S. Europe, W. Asia,
Algeria, Egyjiit. Aerotylus, Fieb. insubricus,
Scop. Mediterranean
Region,
W. Asia,
Canaries,
Algeria, Egypt. Aerotylus, Fieb. insubricus,
Scop. Mediterranean
Region,
W. Asia,
Canaries,
Algeria, Egypt. patruelis, Sturm. S. Earoije, Africa,
Algeria, Egypt. longipes,
Charp. S. Europe, W
Asia,
E. Africa,
Abyssinia,
Algeria, Canaries, Sokotra. errabundus, Fin. Algeria. patruelis, Sturm. S. Earoije, Africa,
Algeria, Egypt. Epacromia, Fisch. drepem, Lat. S. Europe, Algeria,
Asia Minor, Canaries. thalassina,
Fabr,
Europe,
Egypt,
Madeira. lucasi, Bruun. Algeria. longipes,
Charp. S. Europe, W
Asia,
E. Africa,
Abyssinia,
Algeria, Canaries, Sokotra. errabundus, Fin. Algeria. errabundus, Fin. Algeria. lucasi, Bruun. Algeria. Egnatius, St&l. ccerulans, Krauss. Algeria. Leptoscirtus, Sauss. aviculus, Sauss. Egypt. Savignyi, Sauss. Egypt. LocusnDiE. Chlcehwa, Sauss. Kollari, Schulth. Somali. gracilis, Schulth. Somali. Sphingonotus, Fieb. ccerulans, Linn. Europe, W. Asia,
Egypt, Madeira. azurescens, Ramb. Spain, Egypt,
Algeria, Abyssinia. Sphingonotus, Fieb. ccerulans, Linn. Europe, W. Asia,
Egypt, Madeira. gracilis, Schulth. Somali. Quiroguesia, Pant. Quiroguesia, Pant. Quiroguesia, Pant. notabilis. Walk. Canaries,
Spain,
S.W. Asia. Blanchardiana, Sauss. Somali, etc. azurescens, Ramb. Spain, Egypt,
Algeria, Abyssinia. Clausii, Kitt. S. Russia, Egypt. callosus, Fieb. S. Euroi^e, Algeria,. Syria. (Edaleus, Fieb. flavusy Linn. Europe, Asia, Africa,
(Algeria, etc.). asperus, Brulle. Canaries. granulatiis,
Brull^. Canaries, Al-
geria. Locusta, Linn. inorimtus, Schulth. Somali. danica, Linn., Old World
(all W. Africa,
Egypt,
etc.),
Canaries,
Madeira, and Azores. migratoroides,
Reiche. Africa
(Abyssinia, etc.). Eeteropternis, St&l. Savignyi, Krauss. Egypt. Pycnodictya, St&l. Galinieri, Reiche. Abyssinia. Forbesi^ Burr. Sokotra. Locusta, Linn. inorimtus, Schulth. Somali. danica, Linn., Old World
(all W. Africa,
Egypt,
etc.),
Canaries,
Madeira, and Azores. migratoroides,
Reiche. Africa
(Abyssinia, etc.). E t
t
i Sefras, Fin. Algeria. Savignyi, Sauss. Egypt, Nubia. Canariensis,
Sauss. Canaries,
etc. etc. arenarius, Luc. Algeria. niloticus, Sauss. Egypt. latifasciatus. Walk. Egypt, Arabia. tricinctus, W. Egypt, Arabia. octo/asciatus, Serv. Egypt. variegatus. Walk. Egypt. arenarius, Luc. Algeria. niloticus, Sauss. Egypt. Eeteropternis, St&l. Savignyi, Krauss. Egypt. latifasciatus. Walk. Egypt, Arabia. Pycnodictya, St&l. Galinieri, Reiche. Abyssinia. Forbesi^ Burr. Sokotra. octo/asciatus, Serv. Egypt. variegatus. Walk. Egypt. Appendix. 183 Fuiotia, Bona. spinicollis, Borni. Tunis. Ocnerodes, Brunn. Durietii, Bol. Morocco. mtcroptera, Bris. Algeria. nigropunctatus, Luc. Algeria. Volxemi, Bol. Algeria. lo7igicor7iis, Bol. Algeria. Acinipe, Ramb. hispanica, Ramb. Spain, Algeria. Saharse, Pict. Biskra. Muelleri, Krauss. Algeria. Foreli, Pict. Gabes. Algerica, Brunn. Algeria. expama, Brunn. Gibraltar, Algeria,
Mauritanica, Bol. Morocco. Fuiotia, Bona. spinicollis, Borni. Tunis. Ocnerodes, Brunn. Durietii, Bol. Morocco. mtcroptera, Bris. Algeria. nigropunctatus, Luc. Algeria. Volxemi, Bol. Algeria. lo7igicor7iis, Bol. Algeria. Acinipe, Ramb. hispanica, Ramb. Spain, Algeria. Saharse, Pict. Biskra. Muelleri, Krauss. Algeria. Foreli, Pict. Gabes. Algerica, Brunn. Algeria. expama, Brunn. Gibraltar, Algeria,
Mauritanica, Bol. Morocco. Leptopttrnis, Sauss. Rhamses, Sauss. Egypt. canescens, Sauss. Egypt. JJelioscirtus, Sauss. capitanvs, Bonn. Tunis. Finotianus, Sauss. Algeria. Eremoehd^. lo7igicor7iis, Bol. Algeria. Eremobla, Serv. cisti, Fabr. S.W. Europe, Algeria. ClavelU, Luc. Syria, Tunis. pulchripennis, Serv. Egypt. continuata, Serv. Cairo. Acinipe, Ramb. hispanica, Ramb. Spain, Algeria. Saharse, Pict. Biskra. Muelleri, Krauss. Algeria. Foreli, Pict. Gabes. Erernoclioris, Sauss. insignis, Luc. Algeria. Algerica, Brunn. Algeria. expama, Brunn. Gibraltar, Algeria, PrRGOMOBPHID^. Eunupius, Stal. Brunneri, St. Algeria. sitifense, Bris. Algeria. Numidx, Sauss. Tunis. Maroccanus, Sauss. Morocco. granosus, Stal. Algeria. qtutdridentata, Bris. Algeria. Vaucherianus, Sauss. Morocco. Chrotoc/onus, Serv. Bormansi, Bol. Shoa. nngustatus, Blanch. Egypt. Savignyi, Bl. Egypt. Blanchardi, Krauss. Egypt. homolodema, Bl. Sennaar. higuhris, Bl. Egypt. Maroccanus, Sauss. Morocco. Pamphagus, Thunb. elephas, Linn. Algeria. Pyrgomorpha, Serv. conica, Oliv. S. Europe,
Algeria,
Egypt. XipJiocera, Latr. Brunneriana,
Sauss. Somali,
Abyssinia. Egypt. debilis. Fin. Algeria. debilis. Fin. Algeria. Parasphena, Bol. _pic<a, Bol. Massowa. CYETACANTHAORIDiE. Pcecilocera, Serv. hiei-oglyphica, Klug. Dongola. uittata, KI. Dongola. hufonia, Klug. Egypt, Syria. vuhanus, Serv. Egypt. vignaudii, Guer. Abyssinia. Dericorys, Serv. acutispina, Stdl. Egypt. alhidula, Serv. Egypt. Millierii, Borm. Tunis.. Platyphyma, Fisch. ruftpe'i, Brunn. Algeria.. Phymateus, Serv. nUdebrandli, Bol. Somali, Zanzibar. Armindia, Krauss. Brunneri, Kr. Tenerifife. Maura, St&l. apicalis, Bol. Massowa. Xenippa, Stal. aridula, St. Khartum. Dictyophorus, Thuub. griseus, Reiche. Abyssinia. aridula, St. (A'.G^. = Acridium (auct. nee Geofifr.).)^
JEgyptium,
Linn. S. Europe,
N. Africa (Egypt, Algeria, etc.). Schistocerca, Stdl. peregrina,
OLw. S. Europe, W. Asia, N. Africa (Egypt, Algeria,
etc.). Cyrtacanthacris, Walk. compta. Walk. Suakim. (A'.G^. = Acridium (auct. nee Geofifr.).)^
JEgyptium,
Linn. S. Europe,
N. Africa (Egypt, Algeria, etc.). Pamphagodes, Bol. Riffen sis, Bol. Morocco. Pamphagodes, Bol. Riffen sis, Bol. Morocco. Pamphaqid^. Schistocerca, Stdl. peregrina,
OLw. S. Europe, W. Asia, N. Africa (Egypt, Algeria,
etc.). Prionosthenus, Bol. galericulatus, Stal. Egypt. Eurrdgus, Bol. monticohts, Pamb. Cyrtacanthacris, Walk. compta. Walk. Suakim. Appendix. 1 84 llohcckia, Scliulth. ohes'i, Schulth. Somali. Sphodromerus, St&l. decoloratus, Fin. Algeria. inconsjncuus, Schulth. Somali. serapis, Serv. Egypt, Sinai. orieutaUs, Schulth. Somali. sijnatun, Walk. Suakim. ffeteraris, Walk. Somali, Schulth. Somali. Tylotropidius, Stal. Somalicus, Schulth. Somali. Calliptamus, Serv. italicus,
Linn. S. Europe,
W. Asia, N. Africa (Algeria, Tunis),
Madeira, Massowa, Sokotra. ictericus, Serv. Spain, Algeria. vulcanius, Krauss. Canaries. discoidalis. Walk. Egypt. imitator, Walk. Egypt, Arabia. similis, Brunn. Egypt, Syria. turhidus, Walk. Egypt. cidcaratus, Stal. Massowa. Thisoicetrus, Brunn. littoralis, Ramb. S. Europe, Egypt,
Nubia, Algeria, Arabia, etc. cmrulescens, StiU. Massowa. grossus, Schulth. Somali. F.uprepocnemls, Fieb. plorans, Charp. S. Europe, Africa,
Asia, Egypt, Algeria. cinerea, Blanch. Teneriife. cidcaratus, Stal. Massowa. morbosus, Serv. Egypt, Arabia. II.
LIST OF AFRICAN TERMITES. (Compiled from Sjostedt's " Monogi-aph " (Svenska Ak. Handlingar,
34 (4) (1900), by Mr. W. F. Kirby.) (Compiled from Sjostedt's " Monogi-aph " (Svenska Ak. Handlin
34 (4) (1900), by Mr. W. F. Kirby.) (Compiled from Sjostedt's " Monogi-aph " (Svenska Ak. Handlingar,
34 (4) (1900), by Mr. W. F. Kirby.) pallidus, Ramb. Mauritius. Voeltzkou'i, Wasm. Madagascar. Ilodotermes, Hag. mossambicus, Hag. Damara, Cape,
Caffraria,
Natal,
Mozambique,
German E. Africa. ochraccus,
Burm. Egypt,
Timis,
Morocco, Persian Gulf. viator, Latr. Cape (Hex River, etc.). Wasmanni,
Sjost. N. Africa
(Wady M'bellem). Ilavilandi, Sharp. ( = mossambicus
see Sjostedt). viarum, Smeathm. Phil. Trans. Ixxi
p. 189 (1781). Sierra Leone. AuriviUii, Sjostedt. Cape (Darling). Calotermes, Hag. flavicoUis, Fabr. Pahearctic Region
Mediterranean
;
Algeria, Egypt. Camerunensis,
Sj. ( = robustus, Sj.). Cameroons. Madagascariensis, Wasm. Nossi
Be,
Durban
(=
Durhanensis,
Hav. :
sec. Sj.). Ilavilandi,
Sj. Cameroons,
Fer-
nando Po, Congo. Howa, Wasm. Madagascar. rryptops,
Sj. Ugalla (Ituri-Fahro)
[? Uganda]. Ilodotermes, Hag. mossambicus, Hag. Damara, Cape,
Caffraria,
Natal,
Mozambique,
German E. Africa. ochraccus,
Burm. Egypt,
Timis,
Morocco, Persian Gulf. viator, Latr. Cape (Hex River, etc.). Wasmanni,
Sjost. N. Africa
(Wady M'bellem). Ilavilandi, Sharp. ( = mossambicus
see Sjostedt). viarum, Smeathm. Phil. Trans. Ixxi
p. 189 (1781). Sierra Leone. AuriviUii, Sjostedt. Cape (Darling). Calotermes, Hag. flavicoUis, Fabr. Pahearctic Region
Mediterranean
;
Algeria, Egypt. Camerunensis,
Sj. ( = robustus, Sj.). Cameroons. Madagascariensis, Wasm. Nossi
Be,
Durban
(=
Durhanensis,
Hav. :
sec. Sj.). Ilavilandi,
Sj. Cameroons,
Fer-
nando Po, Congo. Howa, Wasm. Madagascar. rryptops,
Sj. Ugalla (Ituri-Fahro)
[? Uganda]. Ithinotermes, Hag. putorius, Sj. Cameroons, Fermiudo
Po, Gaboon, Congo. Acanthoterm.es, Sj. acanthothvrax, Sj. Cameroons. militaris. Hag. Togo,
Congo,
Angelo. spiniger, Sj. Congo. viator, Latr. Cape (Hex River, etc.). Ilavilandi, Sharp. ( = mossambicus
see Sjostedt). Termes, Linn. niger, Sj. Cameroons. gratus, Sj. Togo. vitrialatus, Sj. Congo. Goliath,
Sj. Kilimanjaro,
Masai
Land, Dar-es-Salaam, British Cen-
tral Africa. Natalensis,
Hav. (=
termiticola,
Sj.). Liberia, Gold Coast, Togo,
Congo,
Soudan,
Angola,
Caifra,
Natal, N. Transvaal. bellicosits, Smeathm. (cf. infra'). crucifer,
Sj. Sierra
Leone, Togo,
Cameroons, Congo. cavithorax, Sj. Cameroons. luci/agus,
Rossi. Mediterranean,
Algeria, Egypt. viarum, Smeathm. Phil. Trans. Ixxi
p. 189 (1781). Sierra Leone. Ilavilandi,
Sj. Cameroons,
Fer-
nando Po, Congo. crucifer,
Sj. Sierra
Leone, Togo,
Cameroons, Congo. cavithorax, Sj. Cameroons. Howa, Wasm. Madagascar. Howa,
as . adagasca . rryptops,
Sj. Ugalla (Ituri-Fahro)
[? Uganda]. luci/agus,
Rossi. Mediterranean,
Algeria, Egypt. Appendix. 185 ileslructur, Smeathm. (= flavicoUis,
Walk.). Senegal,
Sierra
Leone,
Natal
(?),
Kordofan,
Bahr-el-
Abiad. ganyika, Tangier, Sennaar, Kordo-
fan, Abyssinia. Entermes, Hav. fungifaher, Sj. S. Leone, Cameroons. bilobatus, Hav. Natal. atrox, Smeathm. S. Leone, Cape. macrothorax,
Sj. Gold
Coast,
Cameroons. longiceps,
Sj,
Cameroons. albotarsalis, Sj. Cameroons, Congo. (Compiled from Sjostedt's " Monogi-aph " (Svenska Ak. Handlin
34 (4) (1900), by Mr. W. F. Kirby.) AurivilUi, Sj. Cameroons. lateralis. Walk. Sierra
Leone,
Cameroons. iruncatus, Wasm. Madagascar. arboricola, Sj. Cameroons. mordax, Smeathm. Sierra Leone. pallidipes, Sj. S. Leoiu', Cameroons. Camerunensis,
Sj. Cameroons. Caffrarix, Sj. Caffraria, Natal. lafericius,
Hav. Natal,
Mozam-
bique. iiquaticus, Sj. Tog;o, Cameroous. iiquaticus, Sj. Tog;o, Cameroous. jiiicrops, Sj. Usambara. iiquaticus, Sj. Tog;o, Cameroous. jiiicrops, Sj. Usambara. latialatus, Sj. Congo. jiiicrops, Sj. Usambara. longiceps,
Sj,
Cameroons. albotarsalis, Sj. Cameroons, Congo. AurivilUi, Sj. Cameroons. latialatus, Sj. Congo. latialatus, Sj. Congo. AurivilUi, Sj. Cameroons. capensis,
De
Geer. Gambia
(?)
Cape, Caffraria. lateralis. Walk. Sierra
Leone,
Cameroons. JJuchhohi,
Rj. Liberia,
Fernando
Po, Gaboon. iruncatus, Wasm. Madagascar. arboricola
Sj
Cameroons iruncatus, Wasm. Madagascar. Po, Gaboon. nngustipennis, Sj. Congo. vulgaris, Hav. Natal. angnstatus,
Ramb. Cape,
Natal,
Caffraria. arboricola, Sj. Cameroons. nngustipennis, Sj. Congo. vulgaris, Hav. Natal. nngustipennis, Sj. Congo. mordax, Smeathm. Sierra Leone. angnstatus,
Ramb. Cape,
Natal,
Caffraria. pallidipes, Sj. S. Leoiu', Cameroons. Camerunensis,
Sj. Cameroons. Lilljeborgi, Sj. Cameroons. Lilljeborgi, Sj. Cameroons. nmplus, Sj. Congo. Lilljeborgi, Sj. Cameroons. nmplus, Sj. Congo. Gabonensis,
Sj. (= MuUeri,
Sj.). Gaboon. Sikoras, Wasm. Madagascar. fuscotihialis, Sj. Cameroons, Gaboon. Gabonensis,
Sj. (= MuUeri,
Sj.). Gaboon. subtilis, Wasm. Aldabra,
Mauri-
tius. parvus, Hav. Gold Coa-st, Natal. tiohilis, Sj. Cameroons. b dit'
Hav
Natal tiohilis, Sj. Cameroons. badit'.s, Hav. Natal. simplicidens, Sj. Cameroons. hasidens, Sj. Togo. anidentatus, Wasm. Gold
Coast,
Zanzibar. heterodon, Sj. Cameroons. badit'.s, Hav. Natal. simplicidens, Sj. Cameroons. rectangularis, Sj. Cameroons. hasidens, Sj. Togo. hasidens, Sj. Togo. hastatus, Hav. Cape. anidentatus, Wasm. Gold
Coast,
Zanzibar. socialis, Sj. Gold Coast, Cameroons. capricornis, Wasm. Madagascar. incertus, Hag. Mozambique, Natal. monodon,
Gerst. Mozambique,
Usegabii, Usagara, Transvaal. aqiiat'icus, Sj. Togo, Cameroons. (belUcosus ; synonyms
:
var. Mos-
mmbica. Hag;
subsp. Sansi-
barica, Wasm. ;
fatale,
Fabr. capensiSy
Latr.J;
subhyalinus,
Ramb.;
viator. Walk. ; falciger,
Gerst.)
beUicosus
;
localities
:
Senegal,
Kerrj- Coast,
Sierra Leone, Gold
Coast,
Togo, Cameroons, Congo,
Angola,
Natal,
N. Transvaal,
Dekgoa Bay, Mozambique, Use-
gaba, L^sambara, Zanzibar, Tan- incertus, Hag. Mozambique, Natal. monodon,
Gerst. Mozambique,
Usegabii, Usagara, Transvaal. baculi, Sj. Cameroons. hospes, Sj. Cameroons. trinervius,
Pamb. Pal. Region,
Sierra
Leone,
Congo,
Damara,
Cape, Natal. (belUcosus ; synonyms
:
var. Mos-
mmbica. Hag;
subsp. Sansi-
barica, Wasm. ;
fatale,
Fabr. capensiSy
Latr.J;
subhyalinus,
Ramb.;
viator. Walk. ; falciger,
Gerst.) mauricianus, Ramb. Mauritius. mauricianus, Ramb. Mauritius. togoensis, Sj. Togo. togoensis, Sj. Togo. geminatus, Wasm. Gold Coast. arboruvi, Smeathm. Senegambia arboruvi, Smeathm. Senegambia
Cameroons, Cape. arboruvi, Smeathm. Se ega b a
Cameroons, Cape. laticeps, Wasm. Madagascar. latifrons,
Sj. (Compiled from Sjostedt's " Monogi-aph " (Svenska Ak. Handlin
34 (4) (1900), by Mr. W. F. Kirby.) Togo,
Cameroous,
Fernando Po. chrysopleura, Sj. Cameroons. canaUculatus, Wasm. Madagascar. nigrita, Wasm. Madagascar. 1 86 INDEX. Armadillidium vulgare, 106
Armed Strongyles, 60
Arrowroot pests of West Indies, 141
Arsenate ot lead wash, 28
Aspidiotus ostrexformis, 25
Aspidiotus perniciosus, 23
Atomaria linearis, 8, 11
Atropos divinatoi'ia, 45, 74
Auchmeroyia, 55
Auchmeroyia depressa, 56
Austrian pine, Fissodes notatus on, 116
Aviculae, 176
Avictda crocea, 178
Avicula heteroptera, 178
Avicula macroptera, 178
Avicula margariti/era, 178
Avicula vexillum, 178
B. Balistes mitis, 177
Banana, insects on, in West Indies, 141,
144
Banded
Pine
Weevil, 116;
prevention
and treatment, 117
Bark spots, 23
Barley, affected with smut and
beetles,
80
Barley, smut and insects in, 5, 80
Bathyscia wollastoni, 84, 87
B#che-de-Mer in West Indies, 173
Beech Coccus, 38
Beet Carrion Beetle, 6
Beetle Mites, 77
Benzine, use
of, for furniture pests, 42
for clothes pests, 44
Bihio hortulanus, 11, 22, 91
Bihio marci, 22
Bihionidx, on mangolds, 11; on
fruit,
22
Bisulphide of carbon, use of, 126
Hlick Fly on mangold, 10
Black Wire worm, 49
Bordeaux Mixture, 48
Brown Curiant Scale, 26 Armadillidium vulgare, 106 A. Armed Strongyles, 60 Arrowroot pests of West Indies, 141 Acacia
Wood,
method
of
destroying
insects in, 128 Arsenate ot lead wash, 28 Aspidiotus ostrexformis, 25 Acari at roots of flowers, 129
Agromyza chermivora, 40
Agrotis exclamationis, 7, 83
Agrotis segetis, 7
Allied Bud Moth, 68 Acari at roots of flowers, 129
Agromyza chermivora, 40
Agrotis exclamationis, 7, 83 Aspidiotus perniciosus, 23 Atomaria linearis, 8, 11 Atropos divinatoi'ia, 45, 74 ,
Agrotis segetis, 7 Agrotis segetis, 7 Auchmeroyia, 55 Allied Bud Moth, 68 Auchmeroyia depressa, 56 Allied Spotted Crane Fly, 101 Allspice, Pimento Borer in West Indies,
139 Almonds,
attacked
by
Indian
Meal
Moth, 124 Andira, insect pests of, 141
Anobium domesticum, 41, 128
Anohium paniceum, 41, 45 Andira, insect pests of, 141
Anobium domesticum, 41, 128
Anohium paniceum, 41, 45
Anohmm tessellatum, 41
; in St. Alban's
Cathedral, 122, 123
Anthocoris fusca, 40
Anthomyia radicum, 11
Anthomyidae, 11
Anthonomus pomorum, 18
Antipest sprayer, 26
Antithesia variegana, 68
Ants, destruction of, 31
Aphides, on apple trees, 27 ; on
carrots,
27
; on osiers and willows, 114
Aphis amygdali, 28
Aphis atripUcis, 10
Aphis brassicse, 93
Aphis mali, 27
Apogonia rauca, 144
Apple Blossom Weevil, 18 ; Sucker, 26
Aphis, 27
;
Fruit Fly, 21
Apples,
maggots
in,
20;
maggots
in
imported
apples,
21;
Lisbon,
21;
Scale on, 22, 29
; Bark Louse, 26
; eggs
on apple
trees, 26
; Aphides on apple
trees, 27
;
Sawfly, 20, 128
Area, 173
Araeocerusfasciculatus, attacking coffee-
berries, 137
Ark Shells, edible, in West Indies, 173 Balistes mitis, 177
Banana, insects on, in West Indies, 141,
144 Banded
Pine
Weevil, 116;
prevention
and treatment, 117
Bark spots, 23 Barley, affected with smut and
beetles,
80 Barley, smut and insects in, 5, 80
Bathyscia wollastoni, 84, 87 Beetle Mites, 77 Benzine, use
of, for furniture pests, 42
for clothes pests, 44 Apples,
maggots
in,
20;
maggots
in
imported
apples,
21;
Lisbon,
21;
Scale on, 22, 29
; Bark Louse, 26
; eggs
on apple
trees, 26
; Aphides on apple
trees, 27
;
Sawfly, 20, 128 Bihionidx, on mangolds, 11; on
fruit,
22 Araeocerusfasciculatus, attacking coffee-
berries, 137 Ark Shells, edible, in West Indies, 173 Index. 187 Bryobia pruni, 26
Bud Mites in Black Currant, 18, 78
Bud Moth, 62;
literature
on, 63;
life-
history
of, 65 ;
at
Wisbech,
73
;
at
Hailsham and Swanley, 128
C. A. Cabbage Aphis on Turnips, 93
Cabbage Boot Fly, 34
Cxcilius, 74
Caitophorus salicivorus, 114
Calandra granaria, 46
Calandra oryzas, 46
Calathus cisteloides, 19
Caliperus, 172
CalUphora, 55
Calocoris fulvomaculatus, 31
Canker fungus mistaken for insect work,
29
Carpocapsa pomonella, 20
Carrion Beetles, 6
Carrot Fly, 108
Carrots, Aphides on, 108
Case-making Clothes Moth, 43
Cassava, insects on, in W. Indies, 141
Cattle, Screw Worm in, 132
Caustic Alkali wash, 25
Cayor Fly, 56
Cecidomyia, 38
Cecidomyia heterobia, 38
;
rosaria, 38
salicis,
38,
129
;
saliciperda,
38
salicina, 38
;
terminalis, 38
Cecidomyidae, 37
Centropomus, 172
Centropristes oculatus, 172
Ceratitis in apples, 21
Cereal Pests, 3, 80
Cetonia aurata, 13
Ceylon, Pearl
Fisheries, 174;
abstract
of Report on, 176
Chafer,
Green
Bose,
13;
garden
and
summer, 12
Chafer Larvae, 12
Chafers, 12
Chanks of commerce, 176
Cheimatohia hrumata, 21
OheJone imbricata, 172
Chelone midas, 172
Chermes abietis, 18
Chermes corticalis, 39
Cherry Sawfly, 21, 72
Chincona pests, 140
Chinosol, use of, 123
Chrysanthemum, land bugs on, 30
Chrysomela marginalis, 91
Cigar Beetle, 125
Cinnabar Moth, 15
Cladius viminalis, 37 Clothes Moths,
43, 128
; treatment
for,
44, 126
Clytus arietis, 92
Coccidae, 29 ;
life-history of, 25
Coccinella septem-punctata, 92
Cocoanut
Palm,
insects
on,
in
West
Indies, 140; Melolonthid
larvae
on,
in Ceylon, 144
Cocoa Plant, insects on, in West Indies,
139
Codling Moth, 20
Coffee berries, damaged by Beetles, 137
Collembola in orchid houses, 110
Colorado
Beetle, notes
on,
in England,
87;
First
Beport
on
outbreak
at
Tilbury, 89
; Report of second visit to
Tilbury
re, 90
;
Report on, 1902, 90
insects
.--ent
as,
91
;
reported
at
Hockley, 93;
at South
Benfleet and
Northfleet, 93
Common Crane Fly, 96
Compsomyia, 55, 131
Compsomyia
macellaria
in man, 131
;
in cattle in St. Lucia, 132
Conorhinus rubrnfasciatus, 130
Cornweal, insect pest in, 141
Corn Weevil, 46
Corrosive sublimate, use of, 42
Cotton, insect pests
of, in West Indies,
139
Cow peas, insect pest
in, 141
Crabs in West Indies, 173
Crane Flies, 94
Crioceris asparagi, 92
Cryptococcus fagi, 38
Cupram, prejiaration of, 48
Currant Scale, brown, 26
; white woolly,
23, 129
Cutworms,
7, 83
D. A. Dactylobius citri, 74
Dadylobius destructor, 74
Dactylobius longipinus, 74
Dart Moth, 7
Death Watch, 41, 45, 122, 128
Depluming Scabies, in fowls, 61
Dermatobia, 55
Dermestes lardarius, 45, 125
Diaspis arr,ygdaU, 23
; on plum, 25
Diaspis bromeliae, 135
Diplosis pyrivora, 22, 128
IHplosis violicola, 106
Dipterous larvae in human excreta, 55
Dorypliora decemlineata, 89
Doryphora juncta, 89
Doryphora melanothorax, 89
Doryphora undecemUneata, 89 Clothes Moths,
43, 128
; treatment
for,
44, 126 Clothes Moths,
43, 128
; treatment
for,
44, 126 Bud Moth, 62;
literature
on, 63;
life-
history
of, 65 ;
at
Wisbech,
73
;
at
Hailsham and Swanley, 128 Coccidae, 29 ;
life-history of, 25 Coccinella septem-punctata, 92 Cocoanut
Palm,
insects
on,
in
West
Indies, 140; Melolonthid
larvae
on,
in Ceylon, 144 Bud Moth, 62;
literature
on, 63;
life-
history
of, 65 ;
at
Wisbech,
73
;
at
Hailsham and Swanley, 128 C. Earwio;R, causing annoyance indoors, ll'.f
" Eclair "' sprayer, 26
Eel-worm disease in oats, 3
Eggs on apple trees, 26
Elamnosoma^ 31
Elephant Chanks, 176
Epilachna, 98
Epunda viminalis, 114
Eriocami a Umacina, 21, 72
Eriophyes pyri, 78
Eriophyes ribis, 18, 79
Eriophyes violx, 107
Ernobius mollis, 41
Eucharis Myrrmcise, 31
Eucheliaj'acobese, 16
Exocsetus roberti, 172
P. Felted Beech Coccus, 38
Ficus,
insects pests
of,
in West
Indies,
141
Fiddlewood, insect on, 141
Filariasis in lamhs, 128
Fish and fishing in West Indies, 172
Flying Fish in West Indies, 172
Foreign Office, Reports to, 145
Forficula auricularia, 119
Fowl's eggs, a parasite in, 128
Frit Fly in oats, 4
Fruit
pests,
18;
Bud
Mites
in
black
currant
bushes,
18;
Apple
Blossom
Weevil, 18
; Strawberry
Beetles, 19
Slug Worms, 21
; Maggots
in apples,
20
; Pear Midge, 22
;
Scale and
False
Scale, 22
; Apple Bark Louse, 26
;
in
orchards at Wisbech, 73
Fruit
trees,
infestation
of, by
Winter
Moth, 21
;
a
general
wash
for, 28
winter washing of, 28
Fumigation, with hydrocyanic acid
gas,
33
;
for
subterranean
pests, 32
;
for
Mealy
Bug
under
glass,
112;
with
bisulphide of carbon, 126. Fungoid disease in black currant leaves,
47
Furniture Beetles, 41
; treatment of, 42
Furniture Insects, 45
Furnitiire Mites, 45
G. Galls on osier plants, 37
Garden Chafer, 12
Gas
treatment,
under
glass,
33;
for
scale
134
Geese
Geese
of,
Gener
Geop
Glos
Ga
Glyc
Olyc
Olyc
Olyc
Glyc
Olyc
Olyc
Olyc
Glyc
Glyc
Goat
Goose
Grand
Grass
Grea
Gree
Grou
32
Grunt
Guin
Hsam
Harp
Hawk
Hear
Hedy
Hipp
Hipp
Hippo
Holot
Homa
Hopl
Hops
Hors
Hors
Hous
Hove
Hydr
Hydr
Hydr
Hydr
ind
Ho
Be
Bu
Hyle
13
Hylob
Hylo
Nypo P. Felted Beech Coccus, 38 Felted Beech Coccus, 38
Ficus,
insects pests
of,
in West
Indies,
141
Fiddlewood, insect on, 141
Filariasis in lamhs, 128
Fish and fishing in West Indies, 172
Flying Fish in West Indies, 172
Foreign Office, Reports to, 145
Forficula auricularia, 119
Fowl's eggs, a parasite in, 128
Frit Fly in oats
4 Fish and fishing in West Indies, 172 H. Hsamulon, 172
Harpalus ruficornis, 19
Hawksbill Turtle, 172
Heart and Dart Motli, 7, 83
Hedya ocellana, 62, 73, 128
Hippodamia variegata, 93
Hippo Flies, 144
Hippono'e escnhnta in Barbados,
17.">
Holothurians in West Indies, 173
Homalomyia, 55
Hoplocampa testudinca, 20, 108
Hops, pests of, 81
Horse Chanks, 176
Horse Worms, 60
Household Pests, Acarine, 120
Hover Flies, 79, 114
HydrsRcia micacea on j^wtatoes, 81, 92
Hydrxcia nebris, 83
Hydrxcia nitela, 83
Hydrocyanic acid
gas, fumigation
with,
indoors,
123
;
under
glass,
33
;
for
Household
Mites,
123
;
for
Larder
Beetle,
126;
under
glass
for
Mealy
Bug, 112
Hylesinus piniperda, 135; treatment of,
136
Hylobius, 116
Hylobius dbietis, 116
Nypomeces squfimosus, 138 Fowl's eggs, a parasite in, 128 Frit Fly in oats, 4 Fruit
pests,
18;
Bud
Mites
in
black
currant
bushes,
18;
Apple
Blossom
Weevil, 18
; Strawberry
Beetles, 19
Slug Worms, 21
; Maggots
in apples,
20
; Pear Midge, 22
;
Scale and
False
Scale, 22
; Apple Bark Louse, 26
;
in
orchards at Wisbech, 73 Fruit
trees,
infestation
of, by
Winter
Moth, 21
;
a
general
wash
for, 28
winter washing of, 28 Fruit
trees,
infestation
of, by
Winter
Moth, 21
;
a
general
wash
for, 28
winter washing of, 28 Fumigation, with hydrocyanic acid
gas,
33
;
for
subterranean
pests, 32
;
for
Mealy
Bug
under
glass,
112;
with
bisulphide of carbon, 126. Fungoid disease in black currant leaves,
47 Furniture Beetles, 41
; treatment of, 42 Hydrocyanic acid
gas, fumigation
with,
indoors,
123
;
under
glass,
33
;
for
Household
Mites,
123
;
for
Larder
Beetle,
126;
under
glass
for
Mealy
Bug, 112 Furniture Insects, 45 Furniture Insects, 45 Furnitiire Mites, 45 Furnitiire Mites, 45 E. Earwio;R, causing annoyance indoors, ll'.f
" Eclair "' sprayer, 26
Eel-worm disease in oats, 3
Eggs on apple trees, 26
Elamnosoma^ 31
Elephant Chanks, 176
Epilachna, 98
Epunda viminalis, 114
Eriocami a Umacina, 21, 72
Eriophyes pyri, 78
Eriophyes ribis, 18, 79
Eriophyes violx, 107
Ernobius mollis, 41
Eucharis Myrrmcise, 31
Eucheliaj'acobese, 16
Exocsetus roberti, 172
P. Felted Beech Coccus, 38
Ficus,
insects pests
of,
in West
Indies,
141
Fiddlewood, insect on, 141
Filariasis in lamhs, 128
Fish and fishing in West Indies, 172
Flying Fish in West Indies, 172
Foreign Office, Reports to, 145
Forficula auricularia, 119
Fowl's eggs, a parasite in, 128
Frit Fly in oats, 4
Fruit
pests,
18;
Bud
Mites
in
black
currant
bushes,
18;
Apple
Blossom
Weevil, 18
; Strawberry
Beetles, 19
Slug Worms, 21
; Maggots
in apples,
20
; Pear Midge, 22
;
Scale and
False
Scale, 22
; Apple Bark Louse, 26
;
in
orchards at Wisbech, 73
Fruit
trees,
infestation
of, by
Winter
Moth, 21
;
a
general
wash
for, 28
winter washing of, 28
Fumigation, with hydrocyanic acid
gas,
33
;
for
subterranean
pests, 32
;
for
M
l
B
i Garden Chafer, 12 C. Cocoa Plant, insects on, in West Indies,
139 Codling Moth, 20 Coffee berries, damaged by Beetles, 137 Collembola in orchid houses, 110 Colorado
Beetle, notes
on,
in England,
87;
First
Beport
on
outbreak
at
Tilbury, 89
; Report of second visit to
Tilbury
re, 90
;
Report on, 1902, 90
insects
.--ent
as,
91
;
reported
at
Hockley, 93;
at South
Benfleet and
Northfleet, 93 ex
Geese, African, etc., 54
Geese, domesticated, origin and
varieties
of, 53
General wash for fruit trees, 28
Geophilus longicorm's, 32
Glossina
lonyipalpns,
v. tuchinoidcs,
in
Gambia, 144
Glyciphagus canestrini, 121
Olyciphagus crameri, 121
Olyciphagus cursor, 45, 120
Olyciphagus dispar, 121
Glyciphagus domestictis, 45, 120
Olyciphagus palmifer, 121
Olyciphagus platygaster, 121
Olyciphagus plumigei; 121
Glyciphagus sciurus, 121
Glyciphagus spiuipes, 121
Goat Moth attacking willows, 113
Gooseberry Fungus, 48
Grandilla, insect pest on, 141
Grasses, insect pest of, 141
Grease-banding and Winter Moth, 129
Green Turtle, 172
Ground garden
jiests, 32; treatment
of,
32
Grunts, 172
Guinea Corn, insect ^xist of, 141
H. Hsamulon, 172
Harpalus ruficornis, 19
Hawksbill Turtle, 172
Heart and Dart Motli, 7, 83
Hedya ocellana, 62, 73, 128
Hippodamia variegata, 93
Hippo Flies, 144
Hippono'e escnhnta in Barbados,
17.">
Holothurians in West Indies, 173
Homalomyia, 55
Hoplocampa testudinca, 20, 108
Hops, pests of, 81
Horse Chanks, 176
Horse Worms, 60
Household Pests, Acarine, 120
Hover Flies, 79, 114
HydrsRcia micacea on j^wtatoes, 81, 92
Hydrxcia nebris, 83
Hydrxcia nitela, 83
Hydrocyanic acid
gas, fumigation
with,
indoors,
123
;
under
glass,
33
;
for
Household
Mites,
123
;
for
Larder
Beetle,
126;
under
glass
for
Mealy
Bug, 112
Hylesinus piniperda, 135; treatment of,
136
Hylobius, 116
Hylobius dbietis, 116
Nypomeces squfimosus, 138 Index i88 Geese, African, etc., 54
Geese, domesticated, origin and
varieties
of, 53
General wash for fruit trees, 28
Geophilus longicorm's, 32
Glossina
lonyipalpns,
v. tuchinoidcs,
in
Gambia, 144
Glyciphagus canestrini, 121
Olyciphagus crameri, 121
Olyciphagus cursor, 45, 120
Olyciphagus dispar, 121
Glyciphagus domestictis, 45, 120
Olyciphagus palmifer, 121
Olyciphagus platygaster, 121
Olyciphagus plumigei; 121
Glyciphagus sciurus, 121
Glyciphagus spiuipes, 121
Goat Moth attacking willows, 113
Gooseberry Fungus, 48
Grandilla, insect pest on, 141
Grasses, insect pest of, 141
Grease-banding and Winter Moth, 129
Green Turtle, 172
Ground garden
jiests, 32; treatment
of,
32
Grunts, 172
Guinea Corn, insect ^xist of, 141 E. Gas
treatment,
under
glass,
33;
for
scale, 134 •M. I. I. Indian Corn,
pests
of,
in West
Indies,
141
Indian Meal Moth, 124
Insect Pests of West Indies, 139
Ixodidae on Toads in Pai;i, 144
J. Japanese Fruit Scale, 23
; on pluui, 25
Jersey, insects on vines in, 73
Julidx
in
potatoes,
15;
destroying
plants at Downton Castle, 105
Julm pulchdlus, 33, 84, 86, 105
K. " Knapsack " Sprayer, 26
L. Labia minor, 119
Lachnus viminalis, 114
Lacon murinus, 15
Lady-bird, 7-spotted, 92
Lambs, Fdariasis in, 128
Land
Bug,
on
chrysanthemums,
30
poisonous, 130
Larder Beetle, 45, 125
Lasloderma testacea, 126
Laverna atra, 64
;
at Wisbech, 68
;
at
Hailshani and Swanlev, 128
Leaf-Cutting Bee, 129
Leaflets,
prepared,
for Board
of
Agri-
culture, 50
;
revised and enlarged, 50
Leal Miners in melons, 129
Leaf Weevils, 73
Leather Jackets, 13
Lecanium rihis, 23, 129
; on plum, 26
Leuconostoc, 127
Ligia oceanica, 106
Lime tree, insect pest of, in West Indies,
142
Lobsters, in West Indies, 173
Locusts,
African,
184;
machines
for
catching, 166
; fungus attacking, 167
;
plants
poisonous
to,
168
;
natural
enemies of, 168
Locust Plagues,
in
Sudan,
Report
on,
164
l.ucilia cxsar, 56
Lucilia sericea, 56
Li/gus contaminatus, 30
Lygus
pratensis,
on
chrysanthemums,
30
; treatment of, 30
J/i/perosia, 133 M. Macaw Worm, 56
Maggot Fly, of Natal, 56
Malpighia, insect on, in Antigua, 142
Mammalia, 29
Mango, insects
on, in AVest
Indies, 140,
142
Mangold,
Pigmy
Beetle
on,
8;
Black. Fly
on,
10;
Flies
(Bibionidx)
on,,
11
; Black Wire Worm in, 49
Marguerite Fly, 108
Marine
resourcet^,
of West Indies, 169
Dr. Duerdeu's Report on, 172
Marsh Crane Fly, 94, 98
Maw Worm, 60
Mealy Bug,
74
; fumigation
for,
under
glass, 112
Megachih
WillouglibieUa, 129
Melanocanthus salicis, 114
Melolontha vidgaris, 12, 92
Melolonthidx, 12
Melons, Leaf-miners in, 129
Merodon clavipes, 107
Merodon eqtiestris, 107
Mesoprion, 172
Millepedes, 87, 105
Minor Shoulder Enob Moth, 114
Modiola, 176
Moles, poison for, 29
Moniezia expansa, 49
Monkey-peas, 80
Mosquito annoyance at Blackheath, 56
Mugil, 172
Murex regius, 176
Muscid larva?, attacking roots, 34
Mussel, edible, in West Indies, 173
Mussel Scale, 23, 26, 75
;
further remedy
for,
26
;
life-history
of, 75
;
natural
enemies of, 77
; treatment of, 77
Mustard Beetle, 16
Mustard pests, 16
Myiasis, human, 55
M>riapoda, 15, 105
Mytilaspis
citricola,
at
Monte
Video,
133
Mytilaspis pomorum, 23, 26, 75
Mytilus exmtus, 173
N. J. Japanese Fruit Scale, 23
; on pluui, 25
Jersey, insects on vines in, 73
Julidx
in
potatoes,
15;
destroying
plants at Downton Castle, 105
Julm pulchdlus, 33, 84, 86, 105 Marine
resourcet^,
of West Indies, 169 •M. Napomyza lateralis, 108
Narcissus Fly, 107
Nedria ditissima, 29
Nematus conjugattis, 37
Nematus pavidus, 37
Nyssa zonaria, 21 Indian Corn,
pests
of,
in West
Indies,
141
Indian Meal Moth, 124
Insect Pests of West Indies, 139
Ixodidae on Toads in Pai;i, 144 Indian Corn,
pests
of,
in West
Indies,
141 Mangold,
Pigmy
Beetle
on,
8;
Black. G. g,
Hylesinus piniperda, 135; treatment of,
136 Galls on osier plants, 37
Garden Chafer, 12 Hylobius, 116 Gas
treatment,
under
glass,
33;
for
scale, 134 Index. 189 K. " Knapsack " Sprayer, 26 " Knapsack " Sprayer, 26 190 Pigeon peas (dried), jiest in, 142
Pigmy Mangold Beetle, 8, 11
Pigmy Potato Beetle, 84
Pineapples, scale on, 135
Pine Beetle, 135
; treatment for, 136
Pissodes notatus, 116
Pith
Moth,
68;
at
Wisbech,
73;
at
Hailsham and Swauley, 128; preven-
tive measures, 71
Plodia interpunctella, 124
Plum Maggot, Red, 128
Poison Baits, 7
Polydesmus complanatus, 32, 105
Pomegranate Pests, of West Indies, 140
Pony Flies, 133
Poplar Sawfly, 37
Porcellio scaber, 105
Potatoes, Myriapoda
in,
15, 87;
Wire
Worm
in, 15
; A new
potato
feeder
(Euchelia
jacobex),
16
;
The
Eosy
Rustic {HydrsRcia
micaceci), 81
;
Siu'-
face Larva3, 83
; Pigmy Potato
Beetle,
84
;
Millepedes, attacking, 87
Potato pests, 15, 81
Psila rosx, 108
Psocida?, on vines, 74
Psylla mali, 26
Pterostichus vulgaris, 92
Puccinia pringsheimiana, 48
Pulvinaria ribesii, 33, 129
Purple Apple Weevil, 20
Pyrochroa serraticornis, 92
Fyrulla carnaria, 176
E. Rat
Flea
and connection with
plague,
144
Red Plum Maggot, 128
Red Spider, killed by gas treatment, 33
Resin Wash, 25, 134
Bhizotrogus solstitialis, 12
Bliynchites baccus, 20
Phynchites cupreus, 20
Rice Weevil, 46
Root Crop Pests, 6, 93
Rose Chafer, 13
Rosy Rustic, attacking potatoes, 81, 92
S. St. Alban's Cathedral, Andbium in, 123
San Jose Scale, 23
Sarcophagus magnifica, 55
Sarcophila, fi5
Sarcoptes lasvis, 61
Sarperda carcharias, 36
Sawfly, on poplar, 37
; on willows, 37 Pigeon peas (dried), jiest in, 142
Pigmy Mangold Beetle, 8, 11
Pigmy Potato Beetle, 84
Pineapples, scale on, 135
Pine Beetle, 135
; treatment for, 136
Pissodes notatus, 116
Pith
Moth,
68;
at
Wisbech,
73;
at
Hailsham and Swauley, 128; preven-
tive measures, 71
Plodia interpunctella, 124
Plum Maggot, Red, 128
Poison Baits, 7
Polydesmus complanatus, 32, 105
Pomegranate Pests, of West Indies, 140
Pony Flies, 133
Poplar Sawfly, 37
Porcellio scaber, 105
Potatoes, Myriapoda
in,
15, 87;
Wire
Worm
in, 15
; A new
potato
feeder
(Euchelia
jacobex),
16
;
The
Eosy
Rustic {HydrsRcia
micaceci), 81
;
Siu'-
face Larva3, 83
; Pigmy Potato
Beetle,
84
;
Millepedes, attacking, 87
Potato pests, 15, 81
Psila rosx, 108
Psocida?, on vines, 74
Psylla mali, 26
Pterostichus vulgaris, 92
Puccinia pringsheimiana, 48
Pulvinaria ribesii, 33, 129
Purple Apple Weevil, 20
Pyrochroa serraticornis, 92
Fyrulla carnaria, 176
E Pigeon peas (dried), jiest in, 142 0. 0. " Knapsack " Sprayer, 26 West Indies, 140, 142 Plodia interpunctella, 124 Pomegranate Pests, of West Indies, 140
Pony Flies, 133
Poplar Sawfly, 37
Porcellio scaber, 105 Potatoes, Myriapoda
in,
15, 87;
Wire
Worm
in, 15
; A new
potato
feeder
(Euchelia
jacobex),
16
;
The
Eosy
Rustic {HydrsRcia
micaceci), 81
;
Siu'-
face Larva3, 83
; Pigmy Potato
Beetle,
84
;
Millepedes, attacking, 87
Potato pests, 15, 81
Psila rosx, 108
Psocida?, on vines, 74
Psylla mali, 26
Pterostichus vulgaris, 92
Puccinia pringsheimiana, 48
Pulvinaria ribesii, 33, 129
Purple Apple Weevil, 20
Pyrochroa serraticornis, 92
Fyrulla carnaria, 176 Oyster-shell Bark Louse, 25, 75 Oysters
in West
Indies, 173
;
pearl
in
Ceylon, 176
Oxyuris curvula, 60 PachyrrJiina maculosa, 13, 99
I'achyrrhina quadri/arsa, 13, 101
Palm seeds, pest on, 142
Paraffin emulsion, 25, 27, 28, 134
Paris Green, 7
Peach Aphis, 28
Peach pest, in Wesst Indies, 140
Pear Midge, 22;
at Guestling,'etc., 128
Pear-leaf Blister Mite, 79
Pear Sawfly, 21, 72
Pearl
Fisheries,
Ceylon, 174
;
abstract
of Eeport on, 176
Pecten ziczag, 173
Penthina pruniana, 68
Phaedon betulae, 16
Phalacrus, 5, 80
Phalacrus corruscus, 80
Phalacrus penicillatus, 80
Phalacrus politus, 80
Philoscia Couchii, 106
Phora forniicarum, 31
Phorbia brassicss, 34
Phygalia pilosaria, 21
Phyllobius, 73
Phyllobius viridaris, 73
Phyllopertha horticola, 12
Phyfochoris pabulinus, 30
Phytomyza, 129
Pliytoptid disease, new, in Violas, 106
Pigeon pea bush, pest on, in AVest Indies,
142 E. Rat
Flea
and connection with
plague,
144
Red Plum Maggot, 128
Red Spider, killed by gas treatment, 33
Resin Wash, 25, 134
Bhizotrogus solstitialis, 12
Bliynchites baccus, 20
Phynchites cupreus, 20
Rice Weevil, 46
Root Crop Pests, 6, 93
Rose Chafer, 13
Rosy Rustic, attacking potatoes, 81, 92
S. St. Alban's Cathedral, Andbium in, 123
San Jose Scale, 23
Sarcophagus magnifica, 55
Sarcophila, fi5
Sarcoptes lasvis, 61
Sarperda carcharias, 36
Sawfly, on poplar, 37
; on willows, 37 Rat
Flea
and connection with
plague,
144
Red Plum Maggot, 128
Red Spider, killed by gas treatment, 33
Resin Wash, 25, 134
Bhizotrogus solstitialis, 12
Bliynchites baccus, 20
Phynchites cupreus, 20
Rice Weevil, 46
Root Crop Pests, 6, 93
Rose Chafer, 13
Rosy Rustic, attacking potatoes, 81, 92 " Knapsack " Sprayer, 26 Oats, eehvorm in, 3
Oats, Frit Fly on, 4
Ochromyia, 55
Ochromyia anthropopliaga, 56
Ocyurus chrysurus, 172
Omasens vulgaris, 19
Oniscus asellus, 106
Orange pests. West Indies, 140, 142
Orange
trees
in Monte Video, attacked
by scale, 133
;
pests in Grenada, 142
Orchid houses, insects in, 109
Oribatidx, 11
Orihata gJohata, 11
Orihata orbicularis, 11
Oscinis frit, 4
Oscinis vastator, 4
Osier plants, insect galls on, 37
;
insects
on, 114
Ostrea parasitica, 173
Oyster-shell Bark Louse, 25, 75
Oysters
in West
Indies, 173
;
pearl
in
Ceylon, 176
Oxyuris curvula, 60
PachyrrJiina maculosa, 13, 99
I'achyrrhina quadri/arsa, 13, 101
Palm seeds, pest on, 142
Paraffin emulsion, 25, 27, 28, 134
Paris Green, 7
Peach Aphis, 28
Peach pest, in Wesst Indies, 140
Pear Midge, 22;
at Guestling,'etc., 128
Pear-leaf Blister Mite, 79
Pear Sawfly, 21, 72
Pearl
Fisheries,
Ceylon, 174
;
abstract
of Eeport on, 176
Pecten ziczag, 173
Penthina pruniana, 68
Phaedon betulae, 16
Phalacrus, 5, 80
Phalacrus corruscus, 80
Phalacrus penicillatus, 80
Phalacrus politus, 80
Philoscia Couchii, 106
Phora forniicarum, 31
Phorbia brassicss, 34
Phygalia pilosaria, 21
Phyllobius, 73
Phyllobius viridaris, 73
Phyllopertha horticola, 12
Phyfochoris pabulinus, 30
Phytomyza, 129
Pliytoptid disease, new, in Violas, 106
Pigeon pea bush, pest on, in AVest Indies,
142 0. Oats, eehvorm in, 3
Oats, Frit Fly on, 4
Ochromyia, 55
Ochromyia anthropopliaga, 56
Ocyurus chrysurus, 172
Omasens vulgaris, 19
Oniscus asellus, 106
Orange pests. West Indies, 140, 142
Orange
trees
in Monte Video, attacked
by scale, 133
;
pests in Grenada, 142
Orchid houses, insects in, 109
Oribatidx, 11
Orihata gJohata, 11
Orihata orbicularis, 11
Oscinis frit, 4
Oscinis vastator, 4
Osier plants, insect galls on, 37
;
insects
on, 114
Ostrea parasitica, 173
Oyster-shell Bark Louse, 25, 75
Oysters
in West
Indies, 173
;
pearl
in
Ceylon, 176
Oxyuris curvula, 60 Pigmy Mangold Beetle, 8, 11 Pith
Moth,
68;
at
Wisbech,
73;
at
Hailsham and Swauley, 128; preven-
tive measures, 71 Orange pests. " Knapsack " Sprayer, 26 Labia minor, 119
Lachnus viminalis, 114
Lacon murinus, 15
Lady-bird, 7-spotted, 92
Lambs, Fdariasis in, 128
Land
Bug,
on
chrysanthemums,
30
poisonous, 130
Larder Beetle, 45, 125
Lasloderma testacea, 126
Laverna atra, 64
;
at Wisbech, 68
;
at
Hailshani and Swanlev, 128
Leaf-Cutting Bee, 129
Leaflets,
prepared,
for Board
of
Agri-
culture, 50
;
revised and enlarged, 50
Leal Miners in melons, 129
Leaf Weevils, 73
Leather Jackets, 13
Lecanium rihis, 23, 129
; on plum, 26
Leuconostoc, 127
Ligia oceanica, 106
Lime tree, insect pest of, in West Indies,
142
Lobsters, in West Indies, 173
Locusts,
African,
184;
machines
for
catching, 166
; fungus attacking, 167
;
plants
poisonous
to,
168
;
natural
enemies of, 168
Locust Plagues,
in
Sudan,
Report
on,
164
l.ucilia cxsar, 56
Lucilia sericea, 56
Li/gus contaminatus, 30
Lygus
pratensis,
on
chrysanthemums,
30
; treatment of, 30
J/i/perosia, 133 Lambs, Fdariasis in, 128 Laverna atra, 64
;
at Wisbech, 68
;
at
Hailshani and Swanlev, 128 Leaf-Cutting Bee, 129 Leaflets,
prepared,
for Board
of
Agri-
culture, 50
;
revised and enlarged, 50 Mussel, edible, in West Indies, 173 Leaf Weevils, 73 Leather Jackets, 13 Locusts,
African,
184;
machines
for
catching, 166
; fungus attacking, 167
;
plants
poisonous
to,
168
;
natural
enemies of, 168 Locusts,
African,
184;
machines
for
catching, 166
; fungus attacking, 167
;
plants
poisonous
to,
168
;
natural
enemies of, 168 Locust Plagues,
in
Sudan,
Report
on,
164 Locust Plagues,
in
Sudan,
Report
on,
164 l.ucilia cxsar, 56 Lygus
pratensis,
on
chrysanthemums,
30
; treatment of, 30
J/i/perosia, 133 Index. T. T. Tamarind, pests on, in West Indies, 142
Taniiia, pest on, in West Indies, 142
Tapestry Moth, 43
Tapeworms,
in sheep, 49
;
in
bile duet
of sheep in Transvaal, 144
Telephorus bicolor, 92
Telephorus rusticus, 92
Teredo chlorotica, 144
Teredo dilatata, 143
Teredo dorsalis, 144
Teredo megofara, 143
Teredo navalis, 144
Teredo norvegica, 144
Teredo micivora, 144
Teredos and Canadian timber, 143
Termites, in Sudan, 155
;
Report on, to
Foreign
Office, 158;
damage
caused
by, 159; methods of prevention and
remedies, 160
Tetranychus telarius, 74
Thymol, use of, for Horse Worms, 60
Thysanoptera (Haliday's types), 129
Ticks on toads
in
Para, 144; on swifts,
144
Tinea pellionella, 43
Tinea frapetzella, 43
Tiniola biselliella, 43, 128
Tiptila lateralis, 94, 98
Tipula oleracea, 13, 94, 96
Tipula paludosa, 94, 98
Tipulidaj,
injurious,
13,
94;
natural
enemies
of,
101
;
preventive
and
remedial measures for, 103
Trapping, for Wire Worm, 33
'J'repang, in West Indies, 173
Trichophaga tapetzella, 43
Trigger Fish, 177
Tropidiurus dentatus, 172
Trygon tvarnak, 111
Trypeba pomoneVa, 21
Tsetse-fiy in Gambia, 144
; and
Buffalo,
correspondence, 147
Tuli|) root, in oats, 3
Turbintlla pyrmn, 176
Turnip Moth (Agrotis segetis), 1
Turnips,
Silpha
rugosa
on,
6
;
cater-
pillars on, 7
; Cabbage Aphis on, 93
Turtles in West Indies, trade of, 172
Tylenchus devastatrix, 3 Tamarind, pests on, in West Indies, 142 Taniiia, pest on, in West Indies, 142 Scolytidx, attacking coffee berries, 138 Tapestry Moth, 43 Scolytidx, attacking coffee berries, 138
Screw
Worms,
in human beings, 131
in cattle in St. S. Index. 191 Scale Disease and False Scale amongst
fruit
trees, 22
;
treatment of affected
trees, 134
Scale
Insects,
on
plum
trees,
25
;
on
orange trees at Monte Video, 133
; on
pineapples, 135
; washes for, 25
Schizoneurafodiens, 109
Schizonetira lanuginosa, on elm, 114
Sclerostomum equinum, 60
Sclerostomum rubrum, 60
Sclerostomum tetracanthum, 60
Scolopendridx, 15, 32
Scolytidx, attacking coffee berries, 138
Screw
Worms,
in human beings, 131
in cattle in St. Lucia, 132
Scymnus discoideus, 40
Sea Esgs in Barbados, 173
" Segging," 3
Senegal Fly, 56
Septaria ribis, 47
Sericulture, Works on, 129
Sheep, Tapeworms in, 49, 144
Shrimps in West Indies, 173
Sill<s, 172
Silpha atrata, 6
Silpha opaca, 6
Silpha rugosa on turnips, 6
Siphidie, 6
Siphocoryne caprex, 114
Si rex
Belies in tir trees, 30
Sirex gigas, 36
Sirex fuvencu% 36
Slug Worms, 21
Smut in barley, 5
Snappers, 172
Snook, 172
Solanum melongena,
pests
on,
in West
Indies, 142
" Soldiers and Sailors," 92
Sponges, in We^t Indies, etc., 179
Spotted Crane Fly, 99
Sprayers, 26
Springtails, 110
Spruce Gall Aphis, 118
Stachia geometrica on coffee berries, 138
Steropus mandidus, 19, 92
Stichophus, 173
Stored goods, pests
on, in West
Indies,
142
Strawberry Beetles, 19
Striped-abdomen Crane Fly, 94, 98 " Suran " pearl-oyster pest, 176
Surface larvae, on turnips, 7, 83
Syrphidss, 79, 114
Syrphus grossularim, 79
Syrphus ribesii, 79
Sweet potatoes, pests on, in West Indies,
142 Scale Disease and False Scale amongst
fruit
trees, 22
;
treatment of affected
trees, 134 Scale
Insects,
on
plum
trees,
25
;
on
orange trees at Monte Video, 133
; on
pineapples, 135
; washes for, 25 Sweet potatoes, pests on, in West Indies,
142 Schizoneurafodiens, 109 Schizonetira lanuginosa, on elm, 114 LONDON: PRINTED BY WILLIAM CLOWES AND SONS, LIMITED,
DCKB STREET, STAMFORD STREET,
S.B., AND GREAT WINDMILL STREET,
W. T. Washes,
for
scale
insects,
25
;
general
-wash for fruit trees, 28
Webbing Clothes Moth, 43
Weevils amongst stored corn,
4fi
;
tre«t-
ment of, 47
;
defoliating rubber, 138
;
Leaf, 73
West Indies, insect pests of, 139
White Ants in Sudan, 155
White Grubs in cabbage, 34
; prevention
and treatment of, 34
Willow Aphides, 114
Willow Beetle, 36
Winter Moth, 21
; at Swanley, 128
Willows, attacked by Goat Moth, 113
;
Sawfly larva3 on, 37
;
insects on, 114 T. Yellow Spotted Crane Fly, 13, 09
YelloAv-tail Fish, 172
Zeuzera xscuJi, 128
Zygoneura, 110
Zygoneura sciarina^
11<) Winter washing of fruit trees, 28
Wire Worm in iiotatoes, 15
Wood I-eopard Moth, 128
Wood Lice, killet by gas treatment, 33
;
in hops, 80 ;
in gardens, 105
Woodpeckers, 123
Wood, pests on, in West Indies, 142
Wood Wasps 36
Woolly Pyrol, insect pest on, at Barba-
dos, 142
X. Xestobiuni tessellatum, 41
Xylophaga jimbriata, 144
Xylophaga hipinnata, 144
Y. Yellow Spotted Crane Fly, 13, 09
YelloAv-tail Fish, 172
Zeuzera xscuJi, 128
Zygoneura, 110
Zygoneura sciarina^
11<) Winter washing of fruit trees, 28
Wire Worm in iiotatoes, 15
Wood I-eopard Moth, 128
Wood Lice, killet by gas treatment, 33
;
in hops, 80 ;
in gardens, 105
Woodpeckers, 123
Wood, pests on, in West Indies, 142
Wood Wasps 36
Woolly Pyrol, insect pest on, at Barba-
dos, 142
X. Xestobiuni tessellatum, 41
Xylophaga jimbriata, 144
Xylophaga hipinnata, 144
Y. Yellow Spotted Crane Fly, 13, 09
YelloAv-tail Fish, 172
Zeuzera xscuJi, 128
Zygoneura, 110
Zygoneura sciarina^
11<) Winter washing of fruit trees, 28
Wire Worm in iiotatoes, 15
Wood I-eopard Moth, 128
Wood Lice, killet by gas treatment, 33
;
in hops, 80 ;
in gardens, 105
Woodpeckers, 123
Wood, pests on, in West Indies, 142
Wood Wasps 36
Woolly Pyrol, insect pest on, at Barba-
dos, 142
X. Xestobiuni tessellatum, 41
Xylophaga jimbriata, 144
Xylophaga hipinnata, 144
Y. Yellow Spotted Crane Fly, 13, 09
YelloAv-tail Fish, 172
Zeuzera xscuJi, 128
Zygoneura, 110
Zygoneura sciarina^
11<) VsHIago mtda, t
Ustihiijo jensinii, 5
V. Vines, insects on, in Jer.sey, 73
w. Washes,
for
scale
insects,
25
;
general
-wash for fruit trees, 28
Webbing Clothes Moth, 43
Weevils amongst stored corn,
4fi
;
tre«t-
ment of, 47
;
defoliating rubber, 138
;
Leaf, 73
West Indies, insect pests of, 139
White Ants in Sudan, 155
White Grubs in cabbage, 34
; prevention
and treatment of, 34
Willow Aphides, 114
Willow Beetle, 36
Winter Moth, 21
; at Swanley, 128
Willows, attacked by Goat Moth, 113
;
Sawfly larva3 on, 37
;
insects on, 114 w. T. Lucia, 132
Scymnus discoideus, 40
Sea Esgs in Barbados, 173
" Segging," 3
Senegal Fly, 56
Septaria ribis, 47
Sericulture, Works on, 129
Sheep, Tapeworms in, 49, 144
Shrimps in West Indies, 173
Sill<s, 172
Silpha atrata, 6
Silpha opaca, 6
Silpha rugosa on turnips, 6
Siphidie, 6
Siphocoryne caprex, 114
Si rex
Belies in tir trees, 30
Sirex gigas, 36
Sirex fuvencu% 36
Slug Worms, 21
Smut in barley, 5
Snappers, 172
Snook, 172
Solanum melongena,
pests
on,
in West
Indies, 142
" Soldiers and Sailors," 92
Sponges, in We^t Indies, etc., 179
Spotted Crane Fly, 99
Sprayers, 26
Springtails, 110
Spruce Gall Aphis, 118
Stachia geometrica on coffee berries, 138
Steropus mandidus, 19, 92
Stichophus, 173
Stored goods, pests
on, in West
Indies,
142
Strawberry Beetles, 19
Striped-abdomen Crane Fly, 94, 98
Subcoccinella, 93
Subcoccinella vigintiquatuorpunctata, 93
Subterranean Insects, destruction of, 32
Sudan, Termites in, 158
; Locust plagues
in, 164
Sugar cane, insects injurious to, in West
Indies, 139, 141
Summer Chafer, 12 Teredos and Canadian timber, 143 Subcoccinella vigintiquatuorpunctata, 93 Subterranean Insects, destruction of, 32 Sudan, Termites in, 158
; Locust plagues
in, 164 Sugar cane, insects injurious to, in West
Indies, 139, 141
Summer Chafer, 12 Index. 192 VsHIago mtda, t
Ustihiijo jensinii, 5
V. Vines, insects on, in Jer.sey, 73
w. Washes,
for
scale
insects,
25
;
general
-wash for fruit trees, 28
Webbing Clothes Moth, 43
Weevils amongst stored corn,
4fi
;
tre«t-
ment of, 47
;
defoliating rubber, 138
;
Leaf, 73
West Indies, insect pests of, 139
White Ants in Sudan, 155
White Grubs in cabbage, 34
; prevention
and treatment of, 34
Willow Aphides, 114
Willow Beetle, 36
Winter Moth, 21
; at Swanley, 128
Willows, attacked by Goat Moth, 113
;
Sawfly larva3 on, 37
;
insects on, 114
Winter washing of fruit trees, 28
Wire Worm in iiotatoes, 15
Wood I-eopard Moth, 128
Wood Lice, killet by gas treatment, 33
in hops, 80 ;
in gardens, 105
Woodpeckers, 123
Wood, pests on, in West Indies, 142
Wood Wasps 36
Woolly Pyrol, insect pest on, at Barba
dos, 142
X. Xestobiuni tessellatum, 41
Xylophaga jimbriata, 144
Xylophaga hipinnata, 144
Y. Y. West Indies, insect pests of, 139 White Ants in Sudan, 155 White Grubs in cabbage, 34
; prevention
and treatment of, 34 Willow Aphides, 114 Willow Beetle, 36 Winter Moth, 21
; at Swanley, 128 Willows, attacked by Goat Moth, 113
;
Sawfly larva3 on, 37
;
insects on, 114 SMITHSONIAN INSTITUTION LIBRARIES
3
9088
00775
5564
|
https://openalex.org/W2969823322
|
https://ojs.unsiq.ac.id/index.php/paramurobi/article/download/178/75
|
Indonesian
| null |
ETOS KERJA ISLAM DALAM PENDIDIKAN ISLAM
|
Paramurobi
| 2,018
|
cc-by-sa
| 5,095
|
Jurnal Paramurobi, Vol. 1, No. 1, Januari-Juni 2018 Jurnal Paramurobi, Vol. 1, No. 1, Januari-Juni 2018 Rifqi Muntaqo, Muhammad Khozinul Huda Fakultas Tarbiyah dan Keguruan UNSIQ Jawa Tengah
Jl. KH. Hasyim Asy'ari Km. 03, Wonosobo, Jawa Tengah
rifqimuntaqo@unsiq.ac.id Keyword; Work ethic, Islamic work ethic. And education. Abstract Ethos as understood as two moral nitai as a unity of goodness of action that is always
done or a commendable action done in certain professions. First, the orientation of moral
values as the basis of a person's moral attitude in behaving and acting as part of his
profession and can be a standard that should or should be followed for others and certain
communities of his time. Second, the evidence of practice and the way in which it executes
the hat. Therefore, the evidence that moral values form the basis of the moral attitude of
this commendable act means one on the plane of thought or concept, and on the other
hand lived its implementation in life or in one of its fields. An important work is based
on three dimensions of human consciousness, namely; Dimension Makrifat, dimension
makrifat based on the ability of a person to be able to understand the signs that Allah
has spread as a mercy for his makhluq. Dimensions of Nature, understanding of self-
attitude to establish a goal where the direction of action is taken. And Shari'a Dimension,
its orientation that action is more imprint than just saying (action speaks louder than a
word). Achieve an Islamic work ethic such as having confidence and optimism, free soul
feeling, God's confidence is always in my heart, insightful and profound, competitive and
positive, authoritative, and most importantly the ability to predict the future well. Rifqi Muntaqo, Muhammad Khozinul Huda Abstrak Etos sebagaimana dipahami sebagai dua nitai moral sebagai satu kesatuan kebaikan
tindakan yang selalu dilakukan atau suatu tindakan terpuji dilakukan pada bidang profesi
tertentu. Pertama, orientasi nilai-nilai moral sebagai dasar sikap moral seseorang dalam
bersikap dan bertindak sebagai bagian profesinya dan bisa menjadi standar yang sebaiknya
atau seharusnya diikuti bagi orang lain dan masyarakat tertentu pada masanya. Kedua, bukti
praktik dan cara pelaksanaannya tentang hat itu. Oleh karena itu, bukti bahwa nilai-nilai
moral sebagai dasar sikap moralnya tindakan terpuji ini, berarti di satu pihak berada pada
dataran pikiran atau konsep, serta di pihak lain dihayati pelaksanaannya dalam kehidupan
atau pada salah satu bidangnya. Suatu pekerjaan penting didasarkan pada tiga dimensi
kesadaran yang dimiliki manusia, yaitu; Dimensi Makrifat, dimensi makrifat didasarkan
pada kemampuan seseorang untuk mampu memahami tanda-tanda yang telah ditebarkan
Allah SWT sebagai rahmat bagi makhluq-Nya. Dimensi Hakikat, pemahaman sikap diri
untuk menetapkan sebuah tujuan kemana arah tindakan dilangkahkan. Dan Dimensi
Syariat, orientasinya bahwa tindakan lebih membekas daripada sekedar berkata (action
speaks louder than a word). Mencapai etos kerja islami dalam pendidikan diantaranya
memiliki sikap percaya diri dan optimis, perasaan jiwa yang merdeka, keyakinan Allah
always in my heart, berwawasan yang luas dan mendalam, berjiwa kompetitif dan positif,
berwibawa, dan yang terpenting kemampuan memprediksi masa depan dengan baik. Kata Kunci; etos kerja, etos kerja islam, pendidikan. Jurnal Paramurobi, Vol. 1, No. 1, Januari-Juni 2018 5
Franz Magnis Suseno, Berfilsafal Dari Konteks,
(Jakarta: Gramedia, 1992), hlm. 120 A. PENDAHULUAN orangtuanya (industri rumah tangga) di luar
jam sekolah. Mereka diajari arti perjuangan
hidup. Bagaimana seharusnya berjuang untuk
memperoleh uang dan memanfaatkan waktu
sebaik-baiknya. Bagi mereka, yang penting
seberapa banyak bisa menabung, bukan seberapa
besar penghasilan yang diperoleh. Industri
rumah tangga di Cina sangat berkembang dan
justru memperkuat perekonomian Cina. Tidak
ada waktu untuk santai atau membuang-buang
waktu dan mereka berusaha selalu memberi nilai
tambah dalam kehidupannya. Kita sering melihat slogan etos kerja yang
menarik terpampang di dinding kantor atau
perusahaan untuk mengingatkan para pekerjanya
terhadap kewajibannya yaitu memiliki tanggung
jawab
meningkatkan
kinerjanya. Namun
berbeda pada realisasinya, kita bisa saksikan
kiprah mereka dalam melaksanakan tugasnya,
misalnya, pada jam sibuk terdapat beberapa
pekerja sibuk ngobrol, membaca koran, ber-
SMS-an dan bahkan duduk-duduk di kantin. Pendidikan dan pelatihan untuk pengembangan
sumber daya manusia di insitusi sudah sering
dilakukan tetapi belum tampak perubahan sikap
mental secara signifikan. Membangun sikap
mental dan etos kerja perlu waktu panjang dan
kesabaran namun tegas. Dengan ketatnya persaingan hidup di cina,
berdampak pada sikap mental positif pekerja
Cina. Mereka sangat menghargai waktu dan
uang dan itu tercermin pada sikap disiplin,
bekerja secara total, memanfaatkan potensi
diri secara maksimal, bersemangat tinggi, tidak
mudah putus asa, kreatif mencipta, berpendirian
kuat dan bekerja secara efektif dan efisien. Lalu bagaimana dengan etos kerja kita, sebagai
seorang muslim yang sejati?. Jika kita menyelami etos kerja penduduk
di Cina, akan dapat kita peroleh gambaran nyata
bagaimana etos kerja tinggi itu dipraktikkan. Dari sejak masa kanak-kanak atau masa sekolah,
mereka sudah dilibatkan dalam kegiatan bisnis 62 Jurnal Paramurobi, Vol. 1, No. 1, Januari-Juni 2018 ISSN : 2615-5680 62 Etos Kerja Islam Dalam Pendidikan Islam B. PEMBAHASAN Franz Magnis Suseno (ditulis F.M. Suseno) menjelaskan, bahwa terdapat kesamaan
antara sikap moral dengan etos, namun tidak
identik. Kesamaannya terletak pada kemutlakan
sikapnya, sedangkan perbedaannya terletak pada
tekanannya. Sikap moral, menegaskan orientasi
pada norma norma sebagai suatu standar yang
harus diikutinya, sementara etos menegaskan
bahwa sikap itu merupakan sikap yang sudah
mantap dan atau sudah menjadi kebiasaan,
suatu hat yang nyata-nyata mempengaruhi,
dan menentukan bagaimana seseorang atau
sekelompok orang mendekati atau melakukan
sesuatu. Karenanya, istilah "etos" diungkapkan
sebagai semangat dan sikap batin yang tetap
pada seseorang atau sekelompok orang sejauh
di dalamnya termuat tekanan-tekanan moral dan
nilai-nilai moral tertentu.5 1
Tim Penyusun Kamus, Kamus Besar Bahasa
Indonesia, (Jakarta: Balai Pustaka, 1988) hlm. 458
dan hlm. 488.
2
Rahmat Fajri, Etos Kerja dalam Islam dan Kristen,
(Yogyakarta: Pustaka Raja, 2005) hlm. 35.
3
Clifford Geertz, Kebudayaan dan Agama, terj.
Francisco Budi Hardiman, (Yogyakarta: Kanisius,
1992), hlm. 50-52
4
Nurcholish Madjid, Islam Doktrin dan Peradaban,
(Jakarta: Yayasan Wakaf Paramadina, 1992), hlm.
411. 4
Nurcholish Madjid, Islam Doktrin dan Peradaban,
(Jakarta: Yayasan Wakaf Paramadina, 1992), hlm.
411. 1
Tim Penyusun Kamus, Kamus Besar Bahasa
Indonesia, (Jakarta: Balai Pustaka, 1988) hlm. 458
dan hlm. 488.t 2
Rahmat Fajri, Etos Kerja dalam Islam dan Kristen,
(Yogyakarta: Pustaka Raja, 2005) hlm. 35.f 3
Clifford Geertz, Kebudayaan dan Agama, terj.
Francisco Budi Hardiman, (Yogyakarta: Kanisius,
1992), hlm. 50-52 1. Pengertian Etos Kerja Dua kata ini seperti yang didefinisikan
dalam kamus besar bahasa indonesi adalah: etos
diartikan sebagai "pandangan yang khas dari
suatu golongan sosial"; adapun kerja memiliki
arti "perbuatan melakukan sesuatu".1 Kata etos
ini dalam pengertian kamus berasal dari bahasa
yunani (ethos) yang berarti watak atau karakter.2 Adapun Clifford geertz menjelaskan
bahwa etos adalah segi-segi moral dan estetis dari
suatu kebudayaan tertentu yang mengandung
unsur-unsur evaluatif. Etos suatu bangsa adalah
sifat, watak dan kualitas kehidupan mereka. Lebih lenjut dikatakan Geertz, etos adalah sikap
mendasar terhadap diri mereka sendiri dan
terhadap dunia mereka yang direfleksikan dalam
kehidupan.3 Menurut Bertens, "etos" adalah salah
satu kata Yunani yang masuk ke dalam banyak
bahasa (termasuk bahasa Indonesia). Kata
itu menunjukkan ciri-ciri, pandangan, nilai-
nilai yang menandai suatu kelompok atau
seseorang. Menurutnya, dalam concise oxford
dictionary (1974). kata "etos" disifatkan sebagai
characteristic spirit of community, people or
system yang maksudnya, "suasana khas yang
menandai suatu kelompok, seseorang atau
system". Etos menunjukkan kepada suasana
khas yang meliputi kerja atau profesi dan perlu
ditekankan bahwa, kata "suasana" harus dipahami
dalam arti baik secara moral. Karenanya, jika
bicara etos dalam profesi tertentu, mesti sebagai
hat yang terpuji. Sedangkan
Nurcholish
Madjid
menjelaskan bahwa etos adalah: karaktristik
dan sikap, kebiasaan serta kepercayaan, dst.,
yang bersifat khusus tentang seorang individu
atau sekelompok manusia. Dan dari perkataan
"etos" terambil pula perkataan "etika" dan "etis"
yang merujuk kepada makna "akhlaq" atau
yang bersifat "akhlaqi", yailu kualitas esensial
seseorang atau suatu kelompok, termasuk suatu
bangsa. Juga dikatakan bahwa "etos" berarti
jiwa khas suatu kelompok manusia, yang dari
jiwa khas itu berkembang pandangan bangsa
tersebut tentang yang baik dan yang buruk,
yakni etikanya.4 Toto
Tasmara
dalam
bukunya
membudavakan etos kerja islami, menurutnya
etos kerja adalah totalitas kepribadian diri serta
cara mengekspresikan, memandang, meyakini,
dan memberikan sesuatu yang bermakna,
yang mendorong dirinya untuk bertindak dan 63 63 ISSN : 2615-5680 Jurnal Paramurobi, Vol. 1, No. 1, Januari-Juni 2018 Rifqi Muntaqo, Muhammad Khozinul Huda meraih amal yang optimal (high performance). Etos kerja muslim didefenisikan sebagai sikap
kepribadian yang melahirkan keyakinan yang
sangat mendalam bahwa bekerja itu bukan
saja untuk memuliakan dirinya, menampakkan
kemanusiaannya, melainkan juga sebagai suatu
manifestasi dari amal saleh. 6
Daryono, Etos Dagang Orang Jawa, (Yogyakarta:
Pustaka Pelajar, 2007), hlm. 4-5. 7
Achmadi, Islam sebagai paradigma ilmu pendidikan
(Yogyakarta: Aditya media, 1992), hlm. 19. 3. Tiga Dimensi Yang Membentuk Etos Kerja Seseorang yang memiliki etos kerja
adalah mereka yang selalu obsesif atau ingin
berbuat sesuatu yang penuh manfaat (shalih)
yang merupakan bagian dari amanah Allah. Itulah sebabnya, cara pandang kita dalam
melaksanakan suatu pekerjaan harus didasarkan
pada tiga dimensi kesadaran, yaitu; 1. Pengertian Etos Kerja Sehingga bekerja
yang didasarkan pada prinsip-prinsip iman
bukan saja menunjukkan fitrah seorang muslim,
melainkan sekaligus meninggikan martabat
dirinya sebagai hamba Allah yang didera
kerinduan untuk menjadikan dirinya sebagai
sosok yang dapat dipercaya, menampilkan
dirinya
sebagai
manusia
yang
amanah,
menunjukkan sikap pengabdian sebagaimana
firman Allah, "Dan tidak Aku menciptakan
jin dan rianusia melainkan supaya mereka
menyembah-Ku." (QS. Adzaariyat: 56). fitrah manusia serta sumber daya insani yang
ada padanya, menuju terbentuknya manusia
seutuhnya (insan kamil) sesuai dengan norma
Islam.7 Sedangkan Pendidikan ke-Islam-an
atau pendidikan Agama Islam yakni upaya
pendidikan agama Islam atau ajaran Islam
dan nilai-nilanya, agar menjadi jiwa, motivasi
bahkan dapat dikatakan way of life seseorang. Pencapaian tujuan pendidikan agama Islam
lebih diorientasikan pada pembangkitan intuisi
agama (perasaan beragama) dan kesiapan rohani
dalam mencapai pengalaman transendental. Dengan demikian tujuan utamanya bukanlah
sekedar
mengalihkan
pengetahuan
dan
keterampilan (sebagai isi pendidikan), melainkan
lebih merupakan suatu ikhtiar untuk menggugah
fitroh insaniyah (to stir up certain innate
powers), sehingga peserta didik bisa menjadi
penganut atau pemeluk agama yang taat dan
baik (muslim paripurna). Kemudian pendidikan
pada umumnya, bertujuan lebih menitikberatkan
pada pemberian pengetahuan dan ketrampilan
khusus dan secara ketat berhubungan dengan
pertumbuhan serta pemilihan dunia kerja
yang diperlukan dalam masyarakat. Dengan
demikian, pendidikan lebih dititikberatkan pada
pencapaian maksimal kognitif-psikomotorik
dan kurang banyak menyentuh sisi rohani serta
karakteristik kemanusaiaannya. Mencermati
penjelasan
tentang
etos
tersebut berarti, dalam kata etos mengandung
dua nitai moral sebagai satu kesatuan kebaikan
tindakan yang selalu dilakukan atau suatu
tindakan terpuji yang Bering dilakukan pada
bidang profesi tertentu. Pertama, orientasi
nilai-nilai moral sebagai dasar sikap moral
seseorang dalam bersikap dan bertindak sebagai
bagian profesinya dan bisa menjadi standar
yang sebaiknya atau seharusnya diikuti (norma
moral) bagi orang lain dan masyarakat tertentu
pada masanya. Kedua, buk-li praktiknya dan
cara pelaksanaannya tentang hat itu. Eksistensi
nilai-nilai moral sebagai clasar sikap moralnya
tindakan terpuji ini, berarti di satu pihak berada
pada dataran pikiran atau konsep, serta di pihak
lain dihayati pelaksanaannya dalam kehidupan
atau pada salah satu bidangnya.6 Jurnal Paramurobi, Vol. 1, No. 1, Januari-Juni 2018
64 8
Toto Tasmara, Membudayakan Etos Kerja Islami,
(Jakarta: Gema Insani, 2002), hal. 6-7. 9
Toto Tasmara, Membudayakan Etos Kerja Islami,..
hlm. 73-76.
10 Toto Tasmara, Membudayakan Etos Kerja Islami,..
hlm. 105-106. 2. Pendidikan Islam Ikhlas
merupakan suatu bentuk perbuatan atau
pelayanan tanpa ikatan. Seorang muslim
takut
bahwa
sesuatu
pekerjaan
yang c. Ikhlas (memiliki moralitas yang bersih),
salah satu kompetensi moral yang dimiliki
seorang yang berbudaya kerja islami itu
adalah nilai keikhlasan. Ikhlas terambil
dari bahasa arab mempunyai arti; bersih
dan mumi (tidak terkontaminasi). Ikhlas
merupakan suatu bentuk perbuatan atau
pelayanan tanpa ikatan. Seorang muslim
takut
bahwa
sesuatu
pekerjaan
yang c. 2. Pendidikan Islam Istilah pendidikan Islam dan pendidikan
agama Islam. Pendidikan Islam adalah segala
usaha untuk memelihara dan mengembangkan a. Dimensi Makrifat, Dimensi makrifat (aku
tahu) harus dihayati oleh setiap subjek pelaku
kerja sehingga dia mampu mengambil posisi ISSN : 2615-5680 Jurnal Paramurobi, Vol. 1, No. 1, Januari-Juni 2018
64 Etos Kerja Islam Dalam Pendidikan Islam a. Pemanfaatan waktu, salah satu esensi
dan hakikat dari etos kerja adalah cara
seseorang menghayati, memahami dan
merasakan betapa berharganya waktu. AI-Qur'an meminta setiap muslim untuk
memperhatikan
dirinya
dalam
rangka
persiapan menghadapi hari esok. Secara
sangat
sederhana,
salah
satu
bukti
mengaktualisasikan ayat al-Qur'an yang
berkaitan dengan waktu tersebut tampaklah
bahwa setiap muslim adalah manusia yang
senang menyusun jadwal harian, mampu
merencanakan pekerjaan dan programnya,
merealisasikannya,
dan
mengevaluasi
seluruh
kegiatannya.9 yang jelas dalam kedudukannya sebagai
pengemban amanah yaitu berupa pekerjaan. Kita tidak mungkin bekerja optimal kecuali
mengetahui terlebih dahulu "siapa aku
dalam hubunganku dengan pekerjaanku,
apa kelebihankun, apa pula kelemahanku"
dan seterusnya. Pemahaman makrifat ini
akan selalu menumbuhkan semangat untuk
berbuat, berkreasi dan berinovasi. Dimensi
makrifat didasarkan pada kemampuan
seseorang untuk mampu memahami tanda-
tanda (alamat) yang telah ditebarkan Allah
SWT sebagai rahmat bagi makhluq-Nya. Dalam hal ini, al-Qur'an memberikan
motivasi yang sangat kuat kepada kita untuk
mendayagunakan daya nalar agar mampu
membedah tabir rahasia dan potensi alam
semesta.8 b. Hidup berhemat dan efisien, berhemat
disini
diartikan
bukanlah
dikarenakan
ingin menunpukkan kekayaan sehingga
melahirkan sifat kikr individualis, melainkan
karena ada suatu reserve bahwa tidak
selamanya waktu itu berjalan secara lurus,
ada up and down, sehingga berhemat berarti
mengestimasikan apa yang akan terjadi di
masa yang akan datang. Sedangkan efisien
berarti melakukan segala sesuatu secara
benar, tepat dan akurat. Adapun efektivitas
berkaitan dengan tujuan atau menetapkan
hal yang benar. Efisien berarti berkaitan
dengan cara melaksanakan, sedangkan
efektivitas berkaitan dengan arah tujuan.10 b. b. Dimensi Hakikat (aku berharap), Sikap diri
untuk menetapkan sebuah tujuan kemana
arah tindakan dilangkahkan. Setiap pribadi
muslim meyakini bahwa niat atau dorongan
untuk menetapkan cita-cita merupakan ciri
bahwa dirinya hidup. c. Dimensi Syariat (aku berbuat), Pengetahuan
tentang peran dan potensi diri, tujuan, serta
harapan-harapan hendaklah dipraktikkan
dalam bentuk tindakan nyata, yang telah
diyakini kebenarannya. Dikatakan bahwa
tindakan lebih membekas daripada sekedar
berkata (action speaks louder than a word). c. Ikhlas (memiliki moralitas yang bersih),
salah satu kompetensi moral yang dimiliki
seorang yang berbudaya kerja islami itu
adalah nilai keikhlasan. Ikhlas terambil
dari bahasa arab mempunyai arti; bersih
dan mumi (tidak terkontaminasi). ISSN : 2615-5680
13 Toto Tasmara, Membudayakan Etos Kerja Islami,..
hlm. 85-86.
14 Toto Tasmara, Membudayakan Etos Kerja Islami,..
hlm. 86.
15 Toto Tasmara, Membudayakan Etos Kerja Islami,..
hlm. 88. 11 Toto Tasmara, Membudayakan Etos Kerja Islami,..
hlm. 78. 11 Toto Tasmara, Membudayakan Etos Kerja Islami,..
hlm. 78.
12 Toto Tasmara, Membudayakan Etos Kerja Islami,..
hlm. 80-81. 4. Etos Kerja Seorang Muslim Ciri-ciri orang yang mempunyai dan
menghayati etos kerja akan tampak dalam sikap
dan tingkah lakunya yang dilandaskan pada
suatu keyakinan yang sangat mendalam bahwa
bekerja itu ibadah dan berprestasi itu indah. Di bawah ini bebcrapa ciri etos kerja seorang
muslim, yakni; 65 65 ISSN : 2615-5680 65 Jurnal Paramurobi, Vol. 1, No. 1, Januari-Juni 2018 Rifqi Muntaqo, Muhammad Khozinul Huda dilatarbelakangi motivasi atau pamrih selain
melaksanakan amanah, walaupun atas nama
"ikhlas dan Uinta", akan berubah menjadi
komoditas semata-mata. Sikap ikhlas bukan
hanya output dari cars dirinya melayani,
melainkan juga input atau masukan yang
membentuk
kepribadiannya
didasarkan
pada sikap yang bersih.11 kepuasan dari pekerjaannya itu. Adapun ciri-
ciri orang yang berkomitmen antara lain;
siap berkorban demi pemenuhan sasaran
perusanaan yang lebih penting, merasakan
dorongan semangat dalam misi yang
lebih besar, dan menggunakan nilai-nilai
kelompok dalam pengambilan keputusan
dan penjabaran-penjabaran pilihan.13 d. Memiliki
kejujuran,
Imam
al-Qusairi
mengatakan bahwa kata shadiq ‘orang yang
jujur’ berasal dari kata shidq ‘kejujuran’. Kata shiddiq adalah bentuk penekanan
(mubalaghah) dari shadiq dan berarti
orang yang di dominasi oleh kejujuran. Dengan demikian, di dalam jiwa seorang
yang jujur itu terdapat komponen nilai
ruhani yang memantulkan berbagai sikap
yang berpihak kepada kebenaran dan
sikap moral yang terpuji (moral upright). Prilaku yang jujur adalah prilaku yang
diikuti oleh sikap tanggung jawab atas apa
yang diperbuatnya. Jujur pada diri sendiri
berarti pula kesungguhan yang amat sangat
untuk meningkatkan dan mengembangkan
misi dan bentuk keberadaannya untuk
memberikan yang terbaik bagi orang lain.12 d. Memiliki
kejujuran,
Imam
al-Qusairi
mengatakan bahwa kata shadiq ‘orang yang
jujur’ berasal dari kata shidq ‘kejujuran’. Kata shiddiq adalah bentuk penekanan
(mubalaghah) dari shadiq dan berarti
orang yang di dominasi oleh kejujuran. Dengan demikian, di dalam jiwa seorang
yang jujur itu terdapat komponen nilai
ruhani yang memantulkan berbagai sikap
yang berpihak kepada kebenaran dan
sikap moral yang terpuji (moral upright). Prilaku yang jujur adalah prilaku yang
diikuti oleh sikap tanggung jawab atas apa
yang diperbuatnya. Jujur pada diri sendiri
berarti pula kesungguhan yang amat sangat
untuk meningkatkan dan mengembangkan
misi dan bentuk keberadaannya untuk
memberikan yang terbaik bagi orang lain.12 f. Istiqomah/
kuat
pendirian,
pribadi
muslim yang professional dan berakhlaq
memiliki sikap konsisten (dari bahasa
latin; consistere), yaitu kemampuan untuk
bersikap secara taat asas, pantang menyerah
dan mampu mempertahankan prinsip serta
komitmennya walau harus berhadapan
dengan risiko yang membahayakan dirinya. Mereka mampu mengendalikan diri dan
mengelola emosinya secara efektif. 12 Toto Tasmara, Membudayakan Etos Kerja Islami,..
hlm. 80-81. Jurnal Paramurobi, Vol. 1, No. 1, Januari-Juni 2018
66 4. Etos Kerja Seorang Muslim Konsekuen
dan
berani
menghadapi
tantangan, adalah merupakan ciri tersendiri
bagi seorang muslim khususnnya untuk
memiliki
keberanian
menerima
suatu
konsekwensi dari keputusan yang telah
diambil. apabila kita melayani dengan pelayanan
yang berkualitas sehingga orang yang ada
disekitar kita merasakannnya. m. Memiliki harga diri, terdapat dua faktor
pembentuk seseorang menjadi professional
dan berakhlaq, yakni konsep diri dan citra
diri. Konsep diri merupakan rujukan utama
bagi hidup seseorang, citra diri adalah
penilaian atas dirinya sendiri, dan harga diri
yaitu penilaian menyeluruh mengenai diri
sendiri, bagaimana ia menyukai pribadinya,
harga diri mempengaruhi kreativitasnya,
dari sini kemudian muncul harga diri. i. Memiliki sikap percaya diri, sikap ini
melahirkan
kekuatan,
keberanian
dan
tegas dalam bersikap dengan berbagai
konsekuensi sebagai akibat perbuatannya. i. Memiliki sikap percaya diri, sikap ini
melahirkan
kekuatan,
keberanian
dan
tegas dalam bersikap dengan berbagai
konsekuensi sebagai akibat perbuatannya. j. Kreatif, karakteristik orang yang kreatif
antara lain; terbuka (mau mendengar
dan anenerima lebih banyak informasi),
pengendapan (hasil dari keterbukaannya
terhadap rangsangan luar, pengetahuan
dan pengalaman orang lain, membuat
mereka
memiliki
kekayaan
bathin),
reproduksi
(mengeluarkan
kembali
hasil pengalaman dirinya dalam bentuk
kreativitasnya, pada awalnya, mungkin saja
mereka meniru orang lain tetapi kemudian
berusaha menyempumakannya sehingga
menjadi dirinya sendiri), evaluasi, clan
pengembangan
diri. n. Memiliki jiwa kepernimpinan, seorang
pemimpin adalah seorang yang mempunyai
personalitas yang tinggi, tidak segan untuk
menerima kritik, bahkan mengikuti apa
yang terbaik. o. Rerorientasi ke masa depan, menetapkan
dengan jelas dan kemudian mengarahkan
tindakannya kepada tujuan yang telah
ditetapkan,
bukan
hanya
berspekulasi
dengan masa depan. p. Memiliki jiwa wiraswasta, orang yang
memiliki jiwa wiraswasta adalah mereka
yang selalu melihat sudut kehidupan dunia
sebagai peluang. Berpikirnya sangat analitis,
melihat segala sesuatu dalam gambar yang
besar. k. Bertanggungjawab;
adalah
merupakan
sikap dan tindakan dari seseorang di
dalam menerima sesuatu sebagai amanah
dengan penuh rasa cinta, orang tersebut
menunaikannya dalam bentuk pilihan-
pilihan yang melahirkan suatu tindakan
yang prestatif atau selalu berprentasi. Sikap ini terkait dengan bagaimana cara
seseorang tersebut mempertahankan prinsip
dan kemudian bertanggung jawab untuk
melaksanakan prinsip-prinsipnya tersebut
dengan tetap menjaga keseimbangan dan
melahirkan nilai bagi harga dirinya. q. Memiliki insting bertanding (fastabiqul
khoirat), Semangat bertanding dalam segala
lapangan kebajikan dan meraih prestasi
merupakan sisi lain dari citra seorang
muslim yang memiliki semangat jihad. Mana mungkin seseorang bisa berlomba
atau bertanding apabila tidak ada gairah
untuk berlatih, bekerja, bergerak dan
berjuang tanpa kenal lelah dan kegagalan. l. 4. Etos Kerja Seorang Muslim Tetap
teguh pada komitmen, positif, dan tidak
rapuh kendati berhadapan dengan situasi
yang menekan. Sikap konsisten melahirkan
kepercayaan diri yang kuat dan memiliki
integritas serta mampu mengelola stress
dengan tetap penuh gairah.14 f. Mereka mampu mengendalikan diri dan
mengelola emosinya secara efektif. Tetap
teguh pada komitmen, positif, dan tidak
rapuh kendati berhadapan dengan situasi
yang menekan. Sikap konsisten melahirkan
kepercayaan diri yang kuat dan memiliki
integritas serta mampu mengelola stress
dengan tetap penuh gairah.14 g. Berdisiplin, sikap berdisiplin (latin; disciple)
yaitu kemampuan untuk mengendalikan
diri dengan tenang dan tetap taat walaupun
dalam
situasi
yang
sangat
menekan. Pribadi yang berdisiplin sangat berhati-
hati dalam mengelola pekerjaan serta
penuh tanggungjawab dalam memunuhi
kewajibannya. Orientasi dari mata hati
dan profesinya terarah pada hasil yang
akan
diraih
(achievements)
sehingga
mampu menyesuaikan diri dalam situasi
yang menantang. Merekapun memiliki
daya adaptibilitas atau keluwesan untuk
menerima inovasi atau gagasan baru.15 g. Berdisiplin, sikap berdisiplin (latin; disciple)
yaitu kemampuan untuk mengendalikan
diri dengan tenang dan tetap taat walaupun
dalam
situasi
yang
sangat
menekan. Pribadi yang berdisiplin sangat berhati-
hati dalam mengelola pekerjaan serta
penuh tanggungjawab dalam memunuhi
kewajibannya. Orientasi dari mata hati
dan profesinya terarah pada hasil yang
akan
diraih
(achievements)
sehingga
mampu menyesuaikan diri dalam situasi
yang menantang. Merekapun memiliki
daya adaptibilitas atau keluwesan untuk
menerima inovasi atau gagasan baru.15 g. e. Memiliki komitmen (aqidah, aqad dan
i’tiqad),
yang
dimaksudkan
dengan
commitment (dari bahasa latin; committere)
adalah
keyaikanan
yang
mengikat
(aqad) sedemikian kukuhnya sehingga
membelenggu seuruh hati nuraninya dan
kemudian mengerakkan perilaku menuju
arah tertentu yang diyakininya (i’tiqad). Penelitian menunjukkan bahwa pegawai
yang memiliki komitmen tinggi pada
perusahaan merupakan orang yang paling
rendah tingkat stresnya dan dilaporkan
bahwa mereka yang berkomitmen itu
merupakan orang yang paling merasakan e. Memiliki komitmen (aqidah, aqad dan
i’tiqad),
yang
dimaksudkan
dengan
commitment (dari bahasa latin; committere)
adalah
keyaikanan
yang
mengikat
(aqad) sedemikian kukuhnya sehingga
membelenggu seuruh hati nuraninya dan
kemudian mengerakkan perilaku menuju
arah tertentu yang diyakininya (i’tiqad). Penelitian menunjukkan bahwa pegawai
yang memiliki komitmen tinggi pada
perusahaan merupakan orang yang paling
rendah tingkat stresnya dan dilaporkan
bahwa mereka yang berkomitmen itu
merupakan orang yang paling merasakan ISSN : 2615-5680 66 Etos Kerja Islam Dalam Pendidikan Islam h. Konsekuen
dan
berani
menghadapi
tantangan, adalah merupakan ciri tersendiri
bagi seorang muslim khususnnya untuk
memiliki
keberanian
menerima
suatu
konsekwensi dari keputusan yang telah
diambil. h. 4. Etos Kerja Seorang Muslim Mereka bahagia karena melayani atau
menolong, dengan selalu memiliki rasa
ingin melayani dengan ketulusan, bukan
karena tugas dan kewajiban saja melainkan
bahwa terdapat kebahagiaan dalam diri kita r. Mandiri (Independent), jiwa yang merdeka
adalah manifestasi dari sebuah keyakinan
bahwa hanya kepada Allah lah kita meminta 67 67 ISSN : 2615-5680 Jurnal Paramurobi, Vol. 1, No. 1, Januari-Juni 2018 Rifqi Muntaqo, Muhammad Khozinul Huda pertolongan, ini merupakan keyakinan yang
melahirkan sikap kemandirian sehingga kita
mampu untuk terus berinovasi dan kreatif. tidak akan ada satu makhluk pun di muka
bumi ini yang mampu mengubah dirinya
kecuali dirinya sendiri. Betapapun hebatnya
seseorang untuk memberikan motivasi, hal
itu hanyalah sebuah kesia-siaan belaka, bila
pada diri orang tersebut tidak ada keinginan
untuk dimotivasi. s. Kecanduan belajar dan haus mencari ilmu,
sikap orang berilmu adalah cara dirinya
berhadapan dengan lingkungan. Dia kritis
dan mampu melakukan analisis yang tajam
terhadap segala fenomena yang berada
disekitarnya, sehingga dia tidak mudah
terkecoh atau terjebak oleh gejala-gejala
yang tidak didukung oleh persyaratan yang
tepat dan benar (faktual) serta proporsional. s. Kecanduan belajar dan haus mencari ilmu,
sikap orang berilmu adalah cara dirinya
berhadapan dengan lingkungan. Dia kritis
dan mampu melakukan analisis yang tajam
terhadap segala fenomena yang berada
disekitarnya, sehingga dia tidak mudah
terkecoh atau terjebak oleh gejala-gejala
yang tidak didukung oleh persyaratan yang
tepat dan benar (faktual) serta proporsional. 16 Toto
Tasmara,
Etos
Kerja
Pribadi
Muslim
(Yogyakarta:
PT
Karipta,
1994),
hal.
53 Jurnal Paramurobi, Vol. 1, No. 1, Januari-Juni 2018
68 5. Pencapaian etos kerja yang islami16 beretos kerja dengan menyandarkan dirinya
kepada Allah dan berjalan dengan penuh
kesungguhan, karena dia yakin bila dirinya
bersungguh-sungguh,
dengan
perangkat
iman, ilmu, dan amal shalehnya, Allah
pasti akan memberikan berbagai jalan
keberhasilan
baginya. g. Memiliki harga diri. Kepribadian seorang
muslim itu sungguh mengesankan. Dia
menjadi pribadi yangdirindukan karena
setiap tindakannya selalu memberikan
bekas yang mendalam dihati orang lain. Cara dia berbica, bertindak, bahkan cara dia
memperlakukan diri dan orang lain sunguuh
memberikan kesan yang menggetarkan hati. c. Allah always in my heart. Inilah bekal
kehidupan yang paling berharga, sebuah
perasaan yang melahirkan nilai moral yang
luhur, bahwa kemanapun kita berpaling
niscaya
disanalah
ada
wajah
Allah
(albaqarah:115), sehingga kesadaran ini
,menjadi dasar pengawasan melekat pada
diri setiap pribadi muslim. h. Berorientasi ke depan. Demi kesuksesan
seorang Muslim berpikir maju ke depan, dia
tidak takut terhadap rintangan, penderitaan,
ataupun
kegagalan. Baginya,
segala
kepedihan adalah modal untuk merasakan
kebahagiaan dimasa depan. 17 Toto Tasmara, Etos Kerja Pribadi Muslim... hlm. 31. 5. Pencapaian etos kerja yang islami16 a. Percaya diri dan optimis, Dalam khazanah
Islam dikenal kata khusnuzh-zhan yang
merupakan rangkuman dari 2 karakter yakni
optimis dan percaya diri. Orang yang optimis
adalah mereka yang melihat cakrawala
dunia penuh warna-warni dan mendorong
dirinya penuh keberanian. Adapun Su’uzh-
zhan, penuh prasangka negative, atau
pesimistis adalah tipe manusia yang melihat
dunia hanya satu warna, tidak ada jalan
keluar, batas cakrawala semakin sempit bagi
hidupnya sehingga dirinya menjadi peragu,
bahkan pengecut. t. Memiliki semangat perantauan, semangat
perantauan adalah semangat menjelajah
hamparan
bumi,
memetik
hikmah,
mengambil pelajaran dari berbagai peristiwa
budaya manusia. Hal ini dapat menyebabkan
seseorang berwawasan universal, tidak
terperangkap dalam fanatisme sempit dan
apalagi kauvinisme. u. Memperhatikan kesehatan dan gizi, hal
ini penting karena etos kerja sangat terkait
dengan bagaimana cara dirinya memelihara
kesehatan
dan
kebugaran
jasmaninya. Bahwa di dalam tubuh yang kuat terdapat
jiwa yang sehat. OPTIMIS
PESIMIS
Masih punya sepuluh
menit lagi
Tinggal sepuluh menit lagi
Saya akan terus mencoba
Ah, sudahlah menyerah
saja
Selalu jadi pemecah
masalah
Selalu jadi bagian dari
permasalahan
Dalam kesulitan ada
kesempatan
Dalam kesempatan banyak
kesulitan OPTIMIS
PESIMIS
Masih punya sepuluh
menit lagi
Tinggal sepuluh menit lagi
Saya akan terus mencoba
Ah, sudahlah menyerah
saja
Selalu jadi pemecah
masalah
Selalu jadi bagian dari
permasalahan
Dalam kesulitan ada
kesempatan
Dalam kesempatan banyak
kesulitan v. Tangguh dan pantang menyerah, bagian dari
kepribadian muslim yang mampu dan gemar
hidup dalam tantangan (challenge). w. Berorientasi pads produktivitas (prilaku
yang selalu mengarah pada cara kerja yang
efisien). b. Jiwa yang merdeka. Jiwa yang merdeka
dan bertanggung jawab itu akan mampu
bereksperimen, yang mana merupakan
bagian dai kecerdasan ruhaniahnya untuk
memperoleh
berbagai
jawaban
atas
perantauannya dimuka bumi ini. Baginya,
the sky is the limit. Sepanjang dia masih
diberi kemampuan untuk hidup, dia tidak
ingin berkreasi, dan memberikan yang
terbaik. Inilah cirri seorang muslim yang x. Memperkaya
jaringan
silaturahim,
bersilaturahmi berarti membuka peluang
dan sekaligus mengikat simpul¬-simpul
informasi dan menggerakkan kehidupan. Silaturahim tidak saja bernilai ibadah, namun
juga sebagai salah satu ruh pengembangan
dirinya. y. Memiliki semangat perubahan, pribadi yang
memiliki etos kerja sangat radar bahwa 53 ISSN : 2615-5680 Jurnal Paramurobi, Vol. 1, No. 1, Januari-Juni 2018
68
ISSN : 2615-5680 Etos Kerja Islam Dalam Pendidikan Islam beretos kerja dengan menyandarkan dirinya
kepada Allah dan berjalan dengan penuh
kesungguhan, karena dia yakin bila dirinya
bersungguh-sungguh,
dengan
perangkat
iman, ilmu, dan amal shalehnya, Allah
pasti akan memberikan berbagai jalan
keberhasilan
baginya. 6. Problematika yang menghambat etos
kerja d. Berwawasan. Semangat pencarian ilmu
pengetauhuan seharusnya melekat kepada
meereka yang mengaku sebagai seorang
muslim
karena
hanya
dengan
ilmu,
kebenaran akan lebih mudah dipahami. Penguasaan
ilmu
juga
menyebabkan
berbagai inovasi dibidang IPTEK. a. Kesalahan paradigma berpikir terhadap
tindakan a. Kesalahan paradigma berpikir terhadap
tindakan Tanda-tanda kesalahan paradigma kita
tentang berpikir terhadap tindakan adalah suka
pesimis dalam melakukan sesuatu, bermalas-
malasan dalam kerja karena nrimo nasib hidup,
tergelam dalam fatalisme, apalagi hanya sekedar
menunggu dan mengkhayal datangnya rizki
dadakan yang duharapkan akan nonmpol karena
lewat undian, judi, lotre walau dengan istilah
yang lebih modern sekalipun.17 e. Memiliki kemampuan bersaing. Hidup
adalah pertempuran yang maha dahsyat. Musuh kita yang paling nyata adalah
setan dengan segala atributnya. Harus ada
keyakinan kuat dalam diri kita untuk tidak
sejengkalpun kalah dari godaan setan. Misalnya seorang hakim, dengan atas nama
hukum dan keadilan dia mengetok palu
dengan suapan materi dari pelaku bukan
murni keadilan. Setan sehalus sutra merayu
kita maka harus kita lawan dengan sekuat
baja rayuan itu. e. Memiliki kemampuan bersaing. Hidup
adalah pertempuran yang maha dahsyat. Musuh kita yang paling nyata adalah
setan dengan segala atributnya. Harus ada
keyakinan kuat dalam diri kita untuk tidak
sejengkalpun kalah dari godaan setan. Misalnya seorang hakim, dengan atas nama
hukum dan keadilan dia mengetok palu
dengan suapan materi dari pelaku bukan
murni keadilan. Setan sehalus sutra merayu
kita maka harus kita lawan dengan sekuat
baja rayuan itu. Harapan (hope) hanya bisa diraih bila
memenuhi kualitas kepribadian yang secara
metaforis dapat digambarkan dalam rumus:
Quality of your (head + heart + hand) + hard working = hope Harapan (hope) hanya bisa diraih bila
memenuhi kualitas kepribadian yang secara
metaforis dapat digambarkan dalam rumus: b. Kesalahan paradigma beribadah Achmadi, Islam sebagai paradigma ilmu
pendidikan, Yogyakarta: Aditya media,
1992. Umat Islam bukanlah umat yang terpenjara
oleh ibadah ritual, Melainkan sangat terobsesi
untuk mewujudkannya dalam bentuk gerak yang
memberikan rahmat bagi sekitarnya. Umat Islam
harus keluar dari kemandegan (statis), karena
sifat statis dan kehilangan ruh untuk berkreasi
(ijtihad dalam bidang amaliah) merupakan
tanda-tanda kematian. Ibarat meneguk air laut,
kian diteguk terasa kian haus pula rasanya. Islam adalah Agama yang bergerak dinamis
penuh energy, tidak pernah mengenal kamus
berhenti dalam berbuat kebaikan, menggapai
prestasi ilahiah karena tempat perhentian seperti
itu hanyalah kelak diliang lahat nanti. Daryono, Etos Dagang Orang Jawa, Yogyakarta:
Pustaka Pelajar, 2007. Fajri, Rahmat, Etos Kerja dalam Islam dan
Kristen, Yogyakarta: Pustaka Raja, 2005. Geertz, Clifford, Kebudayaan dan Agama, terj. Fransisco Budi Hardiman, Yogyakarta:
Kanisius, 1992. Madjid,
Nurcholish,
Islam
Doktrin
dan
Peradaban, Jakarta: Yayasan Wakaf
Paramadina,
1992. Suseno, Franz Magnis, Berfilsafat Dari Konteks,
Jakarta: Gramedia, 1992. DAFTAR PUSTAKA gawang lawan, tidak ada kata diam. b. Kesalahan paradigma beribadah Quality of your (head + heart + hand) + hard working = hope Quality of your (head + heart + hand) + hard working = hope Dengan demikian, kepastian hidup adalah
gerak, dan gerak itulah yang menunjukan
tanda kebermaknaan dalam hidup. Bekerja
berarti sebuah kegiatan yang dinamis, proaktif. Bagaikan seorang penyerang atau gelandang
didalam permainan sepak bola, dia akan terus
berlari untuk mengejar dan menggiring bola ke f. Berpikir positif. Dalam segala hal, jiwa
yang merdeka bebas pula menentukan
pilihannya. Sebagai seorang muslim, dia
hanya akan memilih dan berpihak pada
segala hal yang positif menurut prinsip-
prinsip keimanannya. Ada perasaan hina
yang menusuk hati bila dirinya terperosok
dalam hal-hal negatif. f. Berpikir positif. Dalam segala hal, jiwa
yang merdeka bebas pula menentukan
pilihannya. Sebagai seorang muslim, dia
hanya akan memilih dan berpihak pada
segala hal yang positif menurut prinsip-
prinsip keimanannya. Ada perasaan hina
yang menusuk hati bila dirinya terperosok
dalam hal-hal negatif. 17 Toto Tasmara, Etos Kerja Pribadi Muslim... hlm. 31. 69 ISSN : 2615-5680 69 Jurnal Paramurobi, Vol. 1, No. 1, Januari-Juni 2018 Rifqi Muntaqo, Muhammad Khozinul Huda Jurnal Paramurobi, Vol. 1, No. 1, Januari-Juni 2018
ISSN : 2615-5680 D. SIMPULAN Yang membedakan semangat kerja dalam
Islam adalah kaitannya dengan nilai serta cara
meraih tujuannya. Bagi seorang muslim bekerja
merupakan kewajiban yang hakiki dalam rangka
menggapai ridha Allah. Sedangkan orang kafir
bermujahadah untuk kesenangan duniawi dan
untuk memuaskan hawa nafsu. Tasmara, Toto, Etos Kerja Pribadi Muslim,
Yogyakarta: PT Karipta, 1994. Tasmara, Toto, Membudayakan Etos Kerja
Islami, Jakarta: Gema Insan, 2002. Tim Penyusun Kamus, Kamus Besar Bahasa
Indonesia, Jakarta: Balai Pustaka, 1988. Hal-hal yang harus dilakukan untuk
mencapai etos kerja islami untuk menunjang
pendidikan islam adalah percaya diri dan
optimis, jiwa yang merdeka, Allah always in
my heart, berwawasan, memiliki kemampuan
bersaing, berpikir positif, memiliki harga diri,
dan berorientasi ke depan. Jurnal Paramurobi, Vol. 1, No. 1, Januari-Juni 2018
0
ISSN : 2615-5680
|
https://openalex.org/W4251971083
|
https://hal.archives-ouvertes.fr/hal-02926817/file/acp-8-3459-2008.pdf
|
English
| null |
Mesoscale inversion: first results from the CERES campaign with synthetic data
| null | 2,007
|
cc-by
| 10,325
|
Mesoscale inversion: first results from the CERES
campaign with synthetic data
T. Lauvaux, M. Uliasz, C. Sarrat, F. Chevallier, Philippe Bousquet, Christine
Lac, K. Davis, Philippe Ciais, A. Denning, P. Rayner Mesoscale inversion: first results from the CERES
campaign with synthetic data
T. Lauvaux, M. Uliasz, C. Sarrat, F. Chevallier, Philippe Bousquet, Christine
Lac, K. Davis, Philippe Ciais, A. Denning, P. Rayner campaign with synthetic data
T. Lauvaux, M. Uliasz, C. Sarrat, F. Chevallier, Philippe Bousquet, Christine
Lac, K. Davis, Philippe Ciais, A. Denning, P. Rayner To cite this version: T. Lauvaux, M. Uliasz, C. Sarrat, F. Chevallier, Philippe Bousquet, et al.. Mesoscale inversion: first
results from the CERES campaign with synthetic data. Atmospheric Chemistry and Physics, 2008, 8
(13), pp.3459-3471. 10.5194/acp-8-3459-2008. hal-02926817 1
Introduction Abstract. We investigate the ability of a mesoscale model
to reconstruct CO2 fluxes at regional scale. Formally, we
estimate the reduction of error for a CO2 flux inversion at
8 km resolution in the South West of France, during four days
of the CarboEurope Regional Experiment Strategy (CERES)
in spring 2005. Measurements from two towers and two
airplanes are available for this campaign. The lagrangian
particle dispersion model LPDM was coupled to the non-
hydrostatic model Meso-NH and integrated in a matrix inver-
sion framework. Impacts of aircraft and tower measurements
are quantified separately and together. We find that the con-
figuration with both towers and aircraft is able to significantly
reduce uncertainties on the 4-day averaged CO2 fluxes over
about half of the 300×300 km2 domain. Most of this reduc-
tion comes from the tower measurements, even though the
impact of aircraft measurements remains noticeable. Imper-
fect knowledge of boundary conditions does not significantly
impact the error reduction for surface fluxes. We test alter-
native strategies to improve the impact of aircraft measure-
ments and find that most information comes from measure-
ments inside the boundary layer. We find that there would be
a large improvement in error reduction if we could improve
our ability to model nocturnal concentrations at tower sites. Construction of a coherent picture of the global carbon cycle,
compatible with all available observations remains an impor-
tant and elusive scientific challenge. The two complementary
approaches are termed bottom-up or top-down. In bottom-
up approaches pointwise estimates of CO2 exchange at the
surface (Baldocchi et al., 2001) are integrated in space and
time or are used to validate and calibrate land surface mod-
els (Krinner et al., 2005) together with satellite retrievals of
surface properties. In top-down approaches, these integrated
fluxes are inferred from their signatures on atmospheric con-
centration after being transported in the atmosphere (Enting,
2002). A coherent description requires that both methods
provide statistically consistent flux estimates. Furthermore,
the uncertainties on the two estimates must be small enough
that each estimate carries meaningful information. Uncer-
tainties on bottom-up estimates increase with spatial scale (as
more extrapolation is required) while uncertainties on top-
down estimates increase with decreasing scale due to the ill-
conditioning of the inverse problem and the smoothing of at-
mospheric transport (Enting, 2002). Mesoscale inversion: first results from the CERES campaign with
synthetic data T. Lauvaux1,2, M. Uliasz3, C. Sarrat2, F. Chevallier1, P. Bousquet1, C. Lac2, K. J. Davis4, P. Ciais1, A. S. Denning3, and
P J R
1 T. Lauvaux1,2, M. Uliasz3, C. Sarrat2, F. Chevallier1, P. Bousquet1, C. Lac2, K. J. Davis4, P. C
P. J. Rayner1 1Laboratoire des Sciences du Climat et de l’Environnement/IPSL,CEA-CNRS-UVSQ, Gif-sur-Yv
2Centre Nationale des recherches M´et´eorologiques, Toulouse, France 1Laboratoire des Sciences du Climat et de l’Environnement/IPSL,CEA-CNRS-UVSQ, Gif-sur-Yvette, France
2Centre Nationale des recherches M´et´eorologiques, Toulouse, France 3Department of Atmospheric Sciences, Colorado State University, Fort Collins, Colorado, USA
4Department of Meteorology, The Pennsylvania State University, University Park, Pennsylvania, USA 3Department of Atmospheric Sciences, Colorado State University, Fort Collins, Colorado, USA
4Department of Meteorology, The Pennsylvania State University, University Park, Pennsylvania, U Received: 25 April 2007 – Published in Atmos. Chem. Phys. Discuss.: 19 July 2007
Revised: 13 February 2008 – Accepted: 7 June 2008 – Published: 2 July 2008 Received: 25 April 2007 – Published in Atmos. Chem. Phys. Discuss.: 19 July 2007
Revised: 13 February 2008 – Accepted: 7 June 2008 – Published: 2 July 2008 HAL Id: hal-02926817
https://hal.science/hal-02926817v1
Submitted on 8 Oct 2020 L’archive ouverte pluridisciplinaire HAL, est
destinée au dépôt et à la diffusion de documents
scientifiques de niveau recherche, publiés ou non,
émanant des établissements d’enseignement et de
recherche français ou étrangers, des laboratoires
publics ou privés. HAL is a multi-disciplinary open access
archive for the deposit and dissemination of sci-
entific research documents, whether they are pub-
lished or not. The documents may come from
teaching and research institutions in France or
abroad, or from public or private research centers. Atmos. Chem. Phys., 8, 3459–3471, 2008
www.atmos-chem-phys.net/8/3459/2008/
© Author(s) 2008. This work is distributed under
the Creative Commons Attribution 3.0 License. Atmospheric
Chemistry
and Physics 2
Description of CERES The potential of measurements is evaluated by their im-
pact on the posterior uncertainty of fluxes (e.g. Gloor et al.,
2000; Rayner and O’Brien, 2001; Law et al., 2003). In prin-
ciple this requires only knowledge of the prior uncertainty
covariances for data and concentrations plus knowledge of
atmospheric transport, i.e. no dependence on data or fluxes
themselves. However there is a requirement for consistency
between the quality of the final simulation (difference be-
tween simulated and observed concentrations) and the data
uncertainty (Michalak et al., 2005). Realistic evaluation of
possible data must therefore consider the ability of the mod-
elling system to simulate it. The paper therefore includes a
simple comparison of prior simulation and observed concen-
trations. During CERES (Dolman et al., 2006), CO2 concentration
measurements on instrumented towers and aircraft were col-
lected during six weeks in May–June 2005. The CERES do-
main includes the pine forest of Les Landes (West) and a
large agricultural area with a mixture of winter and summer
crops, in the south west of France (Fig. 1), from Bordeaux
in the north west to Toulouse in the south east. Compared to
other regional studies involving aircraft (e.g. Stephens et al.,
2000; Gerbig et al., 2003; Filippi et al., 2003), the CERES
domain covers a smaller region of about 300×300 km with
several flights each day. Other experiments on similarly-
sized domains were described by (Uliasz et al., 2005) which
comprised a larger set of instrumented towers but without air-
craft, and, Dolman et al. (2002) where aircraft were used but
without towers. The outline of the paper is as follows: First we briefly re-
view the CERES campaign. Then we introduce the various
elements of the inversion system. We commence the results
with a brief comparison of the a priori simulation and ob-
servations then discuss the constraint afforded by the vari-
ous observations. Finally we investigate various alternative
strategies for airborne and tower sampling. The CERES experiment involved two aircraft measuring
atmospheric CO2 during several Intensive Observation Peri-
ods (IOP), and two towers (Marmande and Biscarosse) mea-
suring atmospheric CO2 continuously (Fig. 1). Both towers
provide CO2 concentrations every 30 min, which we average
hourly. The Biscarosse tower is located 2 km from the At-
lantic shore, in the pine forest of Les Landes, at 50 m height
on a 70 m hill. 1
Introduction The problem is compli-
cated by the difficulties of simulating atmospheric transport
at smaller scales on the continents (Geels et al., 2007; P´erez-
Landa et al., 2007). There have been very few cases with
dense enough observations to test consistency. The CERES
campaign described below (Dolman et al., 2006) is an at-
tempt to do this over a limited domain. This paper describes
the components of a top-down inversion system to estimate
fluxes over the domain of CERES and to evaluate the poten-
tial of various atmospheric measurements to constrain fluxes. Actual inversion estimates will be described in a future paper
once all necessary data is validated. Correspondence to: T. Lauvaux
(thomas.lauvaux@lsce.ipsl.fr) Correspondence to: T. Lauvaux
(thomas.lauvaux@lsce.ipsl.fr) Published by Copernicus Publications on behalf of the European Geosciences Union. Published by Copernicus Publications on behalf of the European Geosciences Union. Published by Copernicus Publications on behalf of the European Geosciences Union. 3460 T. Lauvaux et al.: Mesoscale inversion Fig. 1. Map of the vegetation types and the instrumentation over the CERES domain for the 2005 campaign. Fig. 1. Map of the vegetation types and the instrumentation over the CERES domain for the 2005 campaign Atmos. Chem. Phys., 8, 3459–3471, 2008 2
Description of CERES Marmande is located further inland, at 20 m
height, between the pine forest and the agricultural area to Atmos. Chem. Phys., 8, 3459–3471, 2008 www.atmos-chem-phys.net/8/3459/2008/ T. Lauvaux et al.: Mesoscale inversion 3461 the East. In this paper, we study a 4-day period between
the 23 at 6 p.m. to the 27 of May at 12 p.m. which includes
the second IOP of the campaign (26 and 27 of May 2005). The two aircraft, a Piper-Aztec and an ECO-Dimona, flew
ten times between the 23 and 27 of May during the morning
and the early afternoon, with different transects in the region. We used all the flights in this study. Some flights consist of
a vertical profile above the pine forest; some others corre-
spond to a transect from Biscarosse to Marmande, and also
longer ones from Bordeaux to Toulouse mostly in the plan-
etary boundary layer. CO2 concentrations are measured at
high frequency (1 Hz), then averaged to three-minute periods
as detailed below. ment locations and travel to the surface and the boundaries. Compared to a forward mode, all the particles here are used
to estimate fluxes, which reduces the computational cost of
the simulation. The lagrangian model LPDM was enhanced
to simulate aircraft observations based on the precise trajec-
tory of the airplane estimated by GPS (Global Positioning
System). At each second, 10 particles are released at the po-
sition of the aircraft. A longer integration time would yield
more particles and hence more reliable Lagrangian statistics
but would misrepresent the aircraft trajectory. We use higher
resolution for the aircraft measurement period because the
eventual particle distributions are more sensitive to the ex-
plicitly resolved vertical velocity. The dynamical fields in LPDM are forced by mean hori-
zontal winds (u, v), potential temperature, and turbulent ki-
netic energy (TKE) from MESO-NH. At this resolution (less
than 10 km), turbulent motion corresponds to the closure of
the energy budget at each time step. This scalar is used to
quantify turbulent motion of particles as a pseudo random
velocity. Based on the TKE, wind, and potential tempera-
ture, the lagrangian model diagnoses turbulent vertical veloc-
ity and dissipation of turbulent energy. The off-line coupling
between an Eulerian and a Lagrangian model solves most
of the problems of non-linearity in the advection term at the
mesoscale. 3
Models In order to perform a mesoscale inversion,
the non-
hydrostatic atmospheric mesoscale model MesoNH (Lafore
et al., 1998) was coupled offline to the lagrangian disper-
sion model LPDM (Uliasz, 1994). MesoNH enables us to
simulate atmospheric dynamics at high resolution within the
domain together with high frequency CO2 observations of
the CERES campaign. The Lagrangian model is compu-
tationally efficient enough to allow the multiple backward
tracer calculations required for the inversion. While the res-
olution of mesoscale models improves the simulation (and
hence utility) of observations it comes at the cost of a lim-
ited domain size and limited duration. We use a two-way
nesting with resolutions of 8 km and 2 km with 65 levels to
13 km altitude. We simulated the 27 of May at 2 km reso-
lution which is an intensive period of flights, and included
it in the longer simulation of 23–27 May at 8 km resolu-
tion. The 2-way nesting configuration keeps the consistency
of the dynamics between the different grids, which allows
us to use the particle distributions from different runs to-
gether. These two simulations use the analysed data from
the ECMWF as initial and boundary conditions. Dynamical
fields were saved each 20 min, or five min during the flights,
for the off-line coupling with a lagrangian model. The dy-
namical fields at 2 km resolution allow a more precise de-
scription of the vertical transport during the flights. An inter-
comparison study between different RAMS (Regional Atmo-
spheric Modeling System) model versions, WRF (Weather
Research and Forecasting model), and MesoNH (all cou-
pled with biospheric models and prescribed anthropogenic
emissions) showed their ability to reproduce observed CO2
concentrations measured by aircraft during the CERES cam-
paign (Sarrat et al., 2007). At each timestep (from one to 20 s), particles move with a
velocity interpolated from the dynamical fields of the MESO-
NH simulation (5 or 20 min). The timestep depends on
the TKE, following the discretization described in Thomson
(1987). Each time a particle touches the surface, its posi-
tion and release time are saved. Particles here should not be
considered as individual molecules (lost when touching the
surface) but as an air parcel influenced by CO2 fluxes as it
moves along the ground. The formalism for inferring source-receptor relationships
from particle distributions is described by Seibert et al. (2004). 2
Description of CERES Most of the non-linear processes resolved by the
atmospheric model are attributed to a scalar representing the
velocity of the particles. 3
Models At each time step, the fraction of particles (released
from one receptor at one time) within some volume, gives
the influence of that volume on the receptor. If the volume
includes the surface this will yield the influence of surface
sources. If the volume includes the boundary (sides or top) it
yields the influence of that part of the boundary. The particle
distribution at the beginning of the study period defines the
influence of the initial condition. The influence function is
thus decomposed into three different terms, corresponding to
the initial concentration, the surface source contribution, and
the boundary fluxes (Peylin et al., 2005). The tracer backward transport was simulated here by the
Lagrangian Particle Dispersion Model (LPDM) described by
Uliasz (1994). Particles are released from the receptors in a
“backward in time” mode with the wind fields generated by
the eulerian model MesoNH. In a “backward in time” trans-
port mode, particles are released in LPDM from the measure- Mesoscale inversions face the problem of modelling a
limited domain with potentially high contributions from the
boundary fluxes. Regional inversions at the continental scale
(e.g. Gerbig et al., 2003) use large domains and long study
periods to decrease the impact of the lateral boundary and ini-
tial concentrations. In our study, the time period and domain Atmos. Chem. Phys., 8, 3459–3471, 2008 www.atmos-chem-phys.net/8/3459/2008/ 3462 T. Lauvaux et al.: Mesoscale inversion size are limited by computational cost which forces us to deal
with both explicitly. Concentrations at domain boundaries
are defined by a grid of 1◦by 1◦resolution (as typically used
by atmospheric general circulation models) on the horizontal
and 2 levels on the vertical corresponding to the boundary
layer and the free troposphere. The two levels for the bound-
ary fluxes appear as additional unknowns in the inverse sys-
tem. To separate these layers, we use an averaged height of
the boundary layer, given by the daily mean boundary layer
top. This coarse description of the boundaries has the ad-
vantage of introducing fewer unknowns in the inversion, and
still makes it possible to study the impact of boundaries on
the different receptors. The spatial extent of an aircraft obser-
vation is defined by the integration time for the related obser-
vation. A time interval of 3 min produced the best compro-
mise between the need for sufficient particles to gather good
statistics and the ability to resolve the observed distribution. 3
Models Considering the velocity of the aircraft (about 150 km/h), a
3-min time window corresponds to a receptor of less than
8 km long, which is the resolution of the dynamical fields in
Meso-NH. concentrations at the boundary as described above. Minimiz-
ing the equation with respect to s yields s = s0 + C(S0)JT
JC(s0)JT + C(d)
−1
(d −Js0)
(2) (2) More important for this work is the posterior error covariance
for sources given by the expression: C(s)−1 = C(s0)−1 + JT C(d)−1J
(3) (3) We do not solve s in this paper, but focus on the uncertainties
of s (C(s)) that do not depend on the observations d but only
on their errors C(d) and a prior error covariance C(s0). We
notice in Eq. (3) that the posterior covariance C(s) depends
on the prior covariance C(s0). This dependence will be dis-
cussed for our inversion by doubling the prior uncertainty for
the lateral boundaries, to estimate the impact on the error re-
duction for the surface and the boundaries. The value of the
prior flux error was set to 2 gm−2day−1 for the surface and
4 ppm for the boundaries. We do not solve s in this paper, but focus on the uncertainties
of s (C(s)) that do not depend on the observations d but only
on their errors C(d) and a prior error covariance C(s0). We
notice in Eq. (3) that the posterior covariance C(s) depends
on the prior covariance C(s0). This dependence will be dis-
cussed for our inversion by doubling the prior uncertainty for
the lateral boundaries, to estimate the impact on the error re-
duction for the surface and the boundaries. The value of the
prior flux error was set to 2 gm−2day−1 for the surface and
4 ppm for the boundaries. Concerning the estimation of the observation uncertainty,
we assessed it by the comparison of the model results with
aircraft data during the day, and tower data during day and
night. The largest difference is about 3 ppm on different
flights of the 27 of May. Taking into account the uncertainty
of the LPDM model, and the lack of temporal correlations,
we set this diurnal observation error at 4 ppm. The diurnal
variability of the model error for tower data is shown in the
section 6 over the four days. 4
Inversion The lagrangian model backward simulations provide the ma-
trix of influence functions (frequently called the Jacobian J):
the sensitivity of each observation to each unknown. The size
of the J matrix corresponds to the dimension of the vector
of surface fluxes plus the unknown boundary concentrations. The surface fluxes comprise 90×90 points multiplied by the
time resolution of surface fluxes δT (either 1 for a mean flux
or 2 for a split of day and night as explained below). y
Finally, we define the error reduction as : Finally, we define the error reduction as : r = σ post
σ prior
(4) r = σ post
σ prior (4) where σ(x) is the square root of the diagonal of C(x). σpost
represents the posterior error, and σprior the prior error. Re-
trieving a single mean flux over four days is equivalent to
retrieving fluxes at every time step but assuming perfect tem-
poral correlation for every point. From the study of Cheval-
lier et al. (2006) based on daily CO2 fluxes simulated by a
biosphere model and CO2 flux observations, the time cor-
relation of the differences between modelled and observed
CO2 fluxes is still more than 0.5 after 5 days. Our assump-
tion of a perfect error correlation over four days is therefore
defensible. Concerning the spatial correlation of the prior er-
ror covariance, we assumed uncorrelated flux errors on the
domain, as the weakest constraint for this inversion. Using
a spatial error correlation would lead to assume spatial co-
herences in CO2 flux errors which were not clearly identified
at this scale by Chevallier et al. (2006). We do not expect
our prior fluxes to capture the amplitude of the diurnal cycle. We hence allow separate correction of diurnal and noctur-
nal mean fluxes. The separation is somewhat similar to that
of Zupanski et al. (2007) but separated by time rather than
by process. We also investigate the more optimistic option
where the prior flux captures diurnal variability sufficiently
that we need only correct the daily mean. where σ(x) is the square root of the diagonal of C(x). σpost
represents the posterior error, and σprior the prior error. Re-
trieving a single mean flux over four days is equivalent to
retrieving fluxes at every time step but assuming perfect tem-
poral correlation for every point. From the study of Cheval-
lier et al. Atmos. Chem. Phys., 8, 3459–3471, 2008 5
Experiments paths of the next CERES campaign planned for April and
September 2007. For this, we tested different altitudes of
virtual flights in the boundary layer and tried to infer an opti-
mal height from diagnostics of particle distributions and from
the spatial extent of the influence function. Based on one
flight from the 2005 CERES campaign, we created 12 vir-
tual flights with constant altitudes from 100 to 2500 meters. The horizontal coordinates used for these virtual flights cor-
respond to a long transect from Bordeaux to Toulouse. This
optimization gives a first constraint on aircraft measurement
strategy and on the dependency of spatial extent of the fluxes
influencing the observations to flight altitudes . From Eq. 3, the error reduction depends on the prior and ob-
servation error covariance matrices and the Jacobian. For the
observation error, the model error is the main contribution
in our inversion system, compared to measurement uncer-
tainty, aggregation and representation errors. We estimated,
for each tower, the difference between modelled and mea-
sured atmospheric CO2 concentrations during the four days
using the prior estimates of flux. Additionally, we present the
results from the direct simulation of MesoNH coupled with
ISBA-A-gs to identify any differences between the inverse
linearised transport model and the initial direct simulation
used to generate the influence functions. This comparison
will be used to introduce the temporal structure of the uncer-
tainty and its related impact on the error reduction. In a third experiment, the impact of the altitude of CO2
measurement towers was investigated. The sampling altitude
of near-ground stations continuously measuring CO2 largely
determines the spatial extent of the area influencing the mea-
surements. Tall towers observe CO2 concentrations in the
mixed layer and are able to provide information coming from
larger areas (Davis et al., 2003; Gloor et al., 2001). We mod-
elled here a potential tall tower of 300 m height at the exact
position of the actual Biscarosse tower (real altitude is 50 m). The spatial variation of the surface flux contribution is then
compared to the actual one over the 4-day period. We conducted three experiments with the inverse system. For each of them we optimized CO2 fluxes for the four days
of measurements and for each model pixel at 8 km resolu-
tion. 4
Inversion (2006) based on daily CO2 fluxes simulated by a
biosphere model and CO2 flux observations, the time cor-
relation of the differences between modelled and observed
CO2 fluxes is still more than 0.5 after 5 days. Our assump-
tion of a perfect error correlation over four days is therefore
defensible. Concerning the spatial correlation of the prior er-
ror covariance, we assumed uncorrelated flux errors on the
domain, as the weakest constraint for this inversion. Using
a spatial error correlation would lead to assume spatial co-
herences in CO2 flux errors which were not clearly identified
at this scale by Chevallier et al. (2006). We do not expect
our prior fluxes to capture the amplitude of the diurnal cycle. We hence allow separate correction of diurnal and noctur-
nal mean fluxes. The separation is somewhat similar to that
of Zupanski et al. (2007) but separated by time rather than
by process. We also investigate the more optimistic option
where the prior flux captures diurnal variability sufficiently
that we need only correct the daily mean. The boundary concentrations are divided into two levels
and 5 horizontal grid cells for each side of the domain. The
observations consist of 102 hourly concentration measure-
ments from each tower plus an observation each 3 min from
each flight, i.e. 852 observations for the ten flights. The num-
ber of elements in the J matrix is finally 8140×δT ×102×2
+ 8140×δT ×852. The dimension of J makes it possible to solve the inverse
problem using the classical matrix solution for one averaged
flux per grid cell over the four days or for separate averaged
fluxes for day and night. (e.g. Tarantola, 1987; Enting, 2002). Briefly we minimize a cost function: χ2 = 1
2[(s −s0)T C(s0)−1(s −s0)
+(Js −d)T C(d)−1(Js −d)]
(1) (1) Where s represents the unknown sources we seek, s0 the
a priori source estimate, d the observed data and C(x) the
uncertainty covariance of a vector quantity. s and J include Atmos. Chem. Phys., 8, 3459–3471, 2008 www.atmos-chem-phys.net/8/3459/2008/ 3463 T. Lauvaux et al.: Mesoscale inversion (a)
(b)
Fig. 2. CO2 concentrations observed (solid line), modelled with MesoNH and ISBA-A-gs (dashed line), and modelled with our linearised
transport for the inversion (dotted line), at the tower of (a) Biscarosse and (b) Marmande from the 23 of May at 6 p.m. to the 27 at 12 p.m. (b) (a) (b) (a) Fig. 2. 5
Experiments In the first experiment, we tested the potential of the
observations performed during the CERES campaign to re-
duce uncertainties on CO2 fluxes. This study produces maps
of error reduction over the domain. The error reduction was
mapped to a final resolution at 8 km. The different receptors
are simulated separately during the period. Biscarosse, Mar-
mande, and ten different flights were combined to estimate
the final potential of the method. The last case uses only the
two towers, but we separated the flux into two terms, a diur-
nal and a nocturnal, including the temporal structure of the
observation uncertainty from the previous result. We assess
here the impact of the nocturnal and diurnal observations in
the system considering the previous comparison. 4
Inversion CO2 concentrations observed (solid line), modelled with MesoNH and ISBA-A-gs (dashed line), and modelled with our linearised
transport for the inversion (dotted line), at the tower of (a) Biscarosse and (b) Marmande from the 23 of May at 6 p.m. to the 27 at 12 p.m. 6
Results When using only the flight
from Biscarosse to Marmande (Fig. 6a), the largest error re-
ductions (about 90%) occur for one or two boundary pixels
(100 km per pixel) of the upper western boundary. South of
the trajectory, error reduction is about 5% in a small region of
few tens of km. Another region at the East of the Pyrenees,
also visible in the tower case, shows a reduction of about 1%. The last inversion uses all the different available measure-
ments during the second CERES IOP (Fig. 6c). It shows an
extended error reduction around the towers from 60 to 90%,
but also some regions of larger error reduction in the East
and South of the Pyrenees of 10 to 60%. As in the aircraft
case, a few grid cells of the upper boundary show a reduction
of about 90%, the rest being less than 10%. The error re-
duction at the surface is extended by the different flights, and
increased by 15 to 20%. Even though one flight shows lim-
ited impact (Fig. 6b), the addition of the ten different flights
is noticeable in the final error reduction. The backward in time simulation begins on the 28 at mid-
night and lasts 4 days until the 23 of May at 6pm. At the end
of the afternoon of the 27, a northerly wind prevails at the
Biscarosse tower, parallel to the sea shore. During the 26 and
the 27 of May, a sea breeze starts around noon, affecting the
Biscarosse tower (Fig. 3b). The footprint of the Biscarosse
measurement tower is then localised to the West of the tower,
over the Atlantic Ocean. During the 27, the Marmande tower
is affected by a strong south eastern wind, amplified by a val-
ley effect. This wind, called the “Autan wind”, is generated
by the usual synoptic conditions at this period of the year,
with a low pressure system over the Pyrenees mountains. The
cyclonic system induced by the low pressure forces the air
mass from the mediterranean sea to enter the valley between
the Pyrenees mountains and the Montagne Noire (Fig. 3a). During the previous night, between the 26 and the 27 of May,
the two towers show similar footprints to the south east of the
measurement towers, corresponding to the Autan wind situ-
ation. 6
Results At the Marmande tower, even though the main wind
direction remains SouthEasterly, the plume of particles dur-
ing the 26 of May shows a wave distribution corresponding
to changes in the wind direction. During the 24 and the 25
of May, similar meteorological situations occur alternatively
over the domain. The last setup demonstrates the capacity of the inverse sys-
tem under more limiting assumptions. First, we solved for
two different fluxes corresponding to the daily and the noc-
turnal averaged fluxes. Second, we introduced a large diur-
nal variability in the observation uncertainty C(d), of 4 ppmv
during the day, and 100 ppmv during the night, which corre-
sponds roughly to neglecting the nighttime data. This ex-
treme assumption also illustrates the impact of the nighttime
data in estimating both daytime and nighttime CO2 fluxes. We used the results from the data comparison to estimate the
observation error, showing large values during nighttime at
Marmande tower, generalized here to both towers. The four
different error reduction maps presented in the Fig. 7 show
the error reduction when retrieving the diurnal or nocturnal
fluxes, considering a constant observation uncertainty or a
large diurnal variability in the model error. The spatial ex-
tent of significant error reduction is reduced by the increased
number of unknowns (Fig. 7a and b), and also by the larger
observation uncertainty (Fig. 7c and d). Concerning the cases
using a constant observation error (Fig. 7a and b), the error
reduction for the nocturnal flux shows a larger extension but
the high values (>30%) are reduced to a few pixels around
the towers, compared to the daytime flux error reduction. If we increase the nighttime observation error (Fig. 7c and
d), the error reduction decreases for both fluxes, which im-
plies that the nighttime observations constrain daytime and Figure 3 shows the dominant boundary influence from
the West of the domain for the beginning of the 4-day pe-
riod. The dominant location changes during the other days,
but remains localised, which justifies the fine description of
boundary conditions. Considering the particles released from
Biscarosse, we estimated the time distribution of the parti-
cles reaching the boundaries during the 27 of May (Fig. 5). Within 15 h (backward in time) of their release, 99% of them
left the domain. This result defines the maximum backward
integration time for LPDM. 6
Results During this 4-day period, CO2 concentrations were measured
continuously at the two towers, except for a gap in the data
series of Biscarosse starting the 25 of May at 6 p.m. for about
40 h. The comparison of the prior fluxes transported by our
jacobian with the data shows the diurnal variability of the ob-
servation error, much larger during nighttime than daytime,
especially at the Marmande tower (Fig. 2). The nocturnal The second experiment aimed at optimizing the flight Atmos. Chem. Phys., 8, 3459–3471, 2008 www.atmos-chem-phys.net/8/3459/2008/ 3464 T. Lauvaux et al.: Mesoscale inversion accumulation of the atmospheric CO2 at this site reaches
530 ppmv during the second night, whereas the modelled
concentrations in the direct simulation and for the linearised
transport reach about 430 ppmv and 440 ppmv respectively. During the day, the difference between the observed CO2
concentrations and the linearised transport solution is about
5 ppmv for the two tower measurement sites, consistent with
the previous quantification of the observation uncertainty. At
the biscarosse tower measurement site, the diurnal variabil-
ity of the CO2 concentrations is about 20 ppmv, with an error
of about 10 ppmv compared to the two modelled CO2 con-
centrations. The linearised transport solution shows a large
diurnal cycle up to 25 ppmv, slightly higher than observed at
the tower. Although these errors seem large, (Michalak et al.,
2005) noted that, for statistical coherence, the prior simula-
tion error must be smaller than the observational error plus
the prior flux error projected into concentration space. Here
we use the structure of the error mainly for guidance; it is
clear for example that nighttime data at Marmande would be
hard to use. that affect this observation. It also defines the time period of
observations affected by the initial condition. The synoptic
winds were weak during May 27 (about 3 m/s), which makes
this estimation an upper limit for the other days. For the first experiment, we ran an inversion using only the
two towers, measuring throughout the 4-day period (Fig. 6b). In the vicinity of the towers ( tens of km), error reduction can
reach 90% but decreases rapidly to 50%. In the directions
of the main daily winds, the reduction is about 30% 300 km
from the towers, in a narrow band. On the boundaries, the
reduction of error is less than 1% and almost uniform on
different sides of the domain. 6
Results Beyond this (earlier in time)
there are no more particles in the domain hence no fluxes Atmos. Chem. Phys., 8, 3459–3471, 2008 www.atmos-chem-phys.net/8/3459/2008/ T. Lauvaux et al.: Mesoscale inversion
3465
(a)
(b)
Fig. 3. Distribution of the particles (logarithmic scale) released from Biscarosse (B) and Marmande (M) towers (a) the 27 of May between
6:30 a.m. and 7:30 a.m. (b) the 26 of May between 8:30 p.m.‘and 9:30 p.m. 3465 T. Lauvaux et al.: Mesoscale inversion (b) (a) (b) (a) Fig. 3. Distribution of the particles (logarithmic scale) released from Biscarosse (B) and Marmande (M) towers (a) the 27 of May between
6:30 a.m. and 7:30 a.m. (b) the 26 of May between 8:30 p.m.‘and 9:30 p.m. Fig. 5. Time distribution of the particles leaving the domain, re-
leased in a 400-s interval (equivalent to 200 particles) at Biscarosse
tower. Fig. 4. Number of grid cells with an error reduction of 1% (solid
line), with an error reduction equals or more than 30% (dashed line,
top x-axis) and the averaged value of the error reduction within the
domain (dash-dotted line) depending on the altitude of the virtual
flights. Fig. 5. Time distribution of the particles leaving the domain, re-
leased in a 400-s interval (equivalent to 200 particles) at Biscarosse
tower. altitudes, from 100 m to 2500 m. Three different diagnos-
tics were used to estimate the impact of the height of the
trajectory on the surface contribution (Fig. 4): the number
of grid cells with a number of particles corresponding to an Fig. 4. Number of grid cells with an error reduction of 1% (solid
line), with an error reduction equals or more than 30% (dashed line,
top x-axis) and the averaged value of the error reduction within the
domain (dash-dotted line) depending on the altitude of the virtual
flights. Fig. 5. Time distribution of the particles leaving the domain, re-
leased in a 400-s interval (equivalent to 200 particles) at Biscarosse
tower. Fig. 4. Number of grid cells with an error reduction of 1% (solid
line), with an error reduction equals or more than 30% (dashed line,
top x-axis) and the averaged value of the error reduction within the
domain (dash-dotted line) depending on the altitude of the virtual
flights. altitudes, from 100 m to 2500 m. Three different diagnos-
tics were used to estimate the impact of the height of the
trajectory on the surface contribution (Fig. 7
Discussion for the real Biscarosse tower (50 m above the ground). The
most important region of influence covers a large area be-
tween Biscarosse and Marmande towers, and also 2 narrow
bands due to sea breezes on different days of the experiment. We also see a smaller area at the East of the Pyrenees as
with the previous cases. Using the virtual tall Biscarosse
tower, the spatial extent of surface influence is reduced over-
all (Fig. 8b), especially around the tower, and elsewhere is
not visibly enhanced. For the boundary concentrations, re-
duction remains equivalent at less than 1%. The main dif-
ference is due to a change in nighttime response. Particles
released during nighttime are above the nocturnal boundary
layer, compared to the original Biscarosse tower accumulat-
ing particles in the reduced mixed layer. For the second CERES IOP, estimated error reduction larger
than 30% covers an area of about 200 km in the North-South
direction and 100 km in the eastwest direction. This sug-
gests it is possible to do a meaningful top-down/bottom-up
comparison for the pine forest and part of the agricultural
area. Aircraft data showed a small but noticeable contribu-
tion at the surface, using the same observation error as the
one used for daytime tower data. The comparison from Sar-
rat et al. (2007) showed the relatively smaller observation
error for aircraft data modeling compared to near-ground ob-
servations, which is mainly due to the lower variability of
the CO2 concentrations at higher altitudes. We have already
noted the assumptions about spatial and temporal correla-
tions that underlie these results. Clearly, at the resolution
of a few kilometers, more work is necessary on the topic of
flux error correlation, both in space and in time. This is a case where our choice to solve for night and day
fluxes together has a significant impact. Overall, it seems that
the use of tall towers should be treated carefully in inversions
on small regions. Although they are easier to model, (hence
can use lower data uncertainty) a large part of the observed
variability is associated with long range transport, affecting
the boundary conditions of the limited domain. In our inverse system, boundaries appear to be less well
constrained by observations than surface fluxes. 6
Results Error reduction (%) on surface fluxes over the 4-day period: (a) Piper Aztec flight (b) 2 towers, (c) 2 towers+10 flights. 6
Results 4): the number
of grid cells with a number of particles corresponding to an
error reduction greater than 1%, the highest reductions of er-
ror of each flight, and the averaged error reduction for all
the surface grid cells for each flight. The three different
measures are constant in the boundary layer, and decrease
quickly above it, with almost the same shape. Between 500 m
to 1000 m, the three measures of error reduction vary due to
a change in the horizontal mean wind. A local horizontal
wind shear affects the particle distribution when the simu-
lated aircraft flies in one of the main horizontal winds. The
vertical mixing in the boundary layer is strongly dominant
compared to the horizontal mixing, implying that any flight
measurement at one location within the boundary layer will
be influenced by a similar surface area. nighttime fluxes in the same way. The nocturnal flux is
mostly constrained by the nighttime observations as shown
by the narrow high error reduction area around the tower, but
the diurnal flux is also constrained mostly by the nighttime
observations. During the morning, the low boundary layer
height concentrates the particles near the surface, whereas
the high boundary layer during the afternoon distributes the
particles throughout the column. As both periods are consid-
ered diurnal observations, most of the surface influence hap-
pens during the morning. The surface influence is propagated
(backward in time by transport) so that morning observations
are strongly linked to night time fluxes. The last experiment uses only the Biscarosse tower. Thanks to the high resolution of the transport model, we esti-
mated the area of influence in the case of a tall tower (300 m)
at Biscarosse. Figure 8a shows the particle “touchdowns” The second experiment arises from the first one, where it
appears that aircraft measurements are a weaker constraint
than those from towers. In order to optimize this contribu-
tion, a series of 12 virtual flights was simulated at different Atmos. Chem. Phys., 8, 3459–3471, 2008 www.atmos-chem-phys.net/8/3459/2008/ 3466
T. Lauvaux et al.: Mesoscale i
(a)
(b)
(c)
Fig. 6. Error reduction (%) on surface fluxes over the 4-day period: (a) Piper Aztec flight (b) 2 towers, (c) 2 towers+10 flights. 3466 T. Lauvaux et al.: Mesoscale inversion (a) (b) (b) (a)
(c) (c) Fig. 6. Atmos. Chem. Phys., 8, 3459–3471, 2008 7
Discussion The observa-
tion error was set to 4 ppmv during the day and 100 ppmv
during the night following our comparison (Fig. 2). The
increase of the diurnal observation error would mainly de-
crease the error reduction by few percents but won’t affect
seriously its spatial distribution. For future inversions, the
preliminary analysis of flux model errors, and simulated con-
centrations, will guide the definition of the final state vector,
especially the choice of a daily mean or diurnal and noctur-
nal components. The prior flux correlations will be guided
by analysis of flux tower measurements. The error reduc-
tion presented here would be affected by these additional el-
ements depending on the hypothesis introduced in the inverse
system. Fig. 9. Error reduction (%) on surface fluxes over the 4-day period
for one mean flux, using only daytime tower observations. Fig. 9. Error reduction (%) on surface fluxes over the 4-day period
for one mean flux, using only daytime tower observations. boundaries. For aircraft measurements, the contribution of
the boundaries is more important due to the fact that most of
the particles released at higher altitudes reach the boundaries
without touching the surface. However, even with the use
of aircraft, the optimization of the boundary fluxes remains
limited because aircraft measurements represent many fewer
particles than continuous ground sites. This means also that
the impact of the boundaries on final CO2 concentrations ob-
served is reduced. The prior uncertainty of the boundaries
was increased to assess its impact on the posterior error. The
error reduction for surface fluxes remains constant while high
values at the boundaries show higher reductions. The result
is explained by Eq. (3). For a given C(d) and J there is a
larger reduction of error if C(s0)−1 is small, i.e. a large prior
uncertainty. The lack of impact on the reduction of error for
the surface is explained by the weak coupling between the
surface and boundary parts of the inversion. The behaviour of error reduction is sensitive to vertical ve-
locity in the boundary layer. Averaged vertical velocity es-
timated in a preliminary simulation appeared to be weaker
than observed. A simple first approach to examining the im-
portance of this issue is to estimate the impact of strongly
varying day/night turbulence intensities and depth of mix-
ing. 7
Discussion The final
contribution of boundaries remains limited because particles
released in LPDM touch the surface three times on average
before leaving the domain after passing once through the Atmos. Chem. Phys., 8, 3459–3471, 2008 www.atmos-chem-phys.net/8/3459/2008/ 3467 T. Lauvaux et al.: Mesoscale inversion Fig. 7. Error reduction (%) on surface fluxes over the 4-day period using only the tower measurements: (a) diurnal mean flux, constant
bservation error of 4 ppm (b) nocturnal mean flux, constant observation error of 4 ppm (c) diurnal mean flux, using only daytime data (d)
octurnal mean flux, using only daytime data. Fig. 7. Error reduction (%) on surface fluxes over the 4-day period using only the tower measurements: (a) diurnal mean flux, constant
observation error of 4 ppm (b) nocturnal mean flux, constant observation error of 4 ppm (c) diurnal mean flux, using only daytime data (d)
nocturnal mean flux, using only daytime data. (a)
(b)
Fig. 8. Particle distribution over 4-day period: (a) Real Biscarosse tower (50 m), (b) Fictive tall tower at Biscarosse (300 m). (b) (a) (b) (a) Fig. 8. Particle distribution over 4-day period: (a) Real Biscarosse tower (50 m), (b) Fictive tall tower at Biscarosse (300 m). g. 8. Particle distribution over 4-day period: (a) Real Biscarosse tower (50 m), (b) Fictive tall tower at Biscaro Fig. 8. Particle distribution over 4-day period: (a) Real Biscarosse tower (50 m), (b) Fictive tall tower at Biscarosse (300 m). Atmos. Chem. Phys., 8, 3459–3471, 2008 www.atmos-chem-phys.net/8/3459/2008/ T. Lauvaux et al.: Mesoscale inversion 3468 (Sarrat et al., 2007). The nocturnal accumulation of CO2 in
the boundary layer at Marmande tower remains lower than
observed in the direct simulation (Fig. 2b). The offline cou-
pling with the lagrangian model reinforces this additional
vertical diffusion. The integration of the particles near the
surface is optimal between 50 m to 100 m high during the
night, higher than expected. The reduced height of the noc-
turnal boundary layer explains the diurnal variability. The
absolute error on the boundary layer height during the night
induces a larger relative error compared to daytime, which is
directly related to the modelled CO2 concentration. The er-
ror reduction of the diurnal CO2 fluxes is less affected by the
larger nocturnal observation error than nocturnal CO2 fluxes. But the final error reduction is larger for the nocturnal com-
ponent of the fluxes. 7
Discussion Considering the inversion setup for
one mean flux, the error reduction using the two towers of
the campaign is reduced but still covers a few tenths of km
(Fig. 9). We suppose in this experiment the model is able to
reproduce the diurnal variability of the CO2 fluxes, as shown
in the study of Ahmadov et al. (2007) during the same cam-
paign using the coupled model WRF-VPRM. The observa-
tion error was set to 4 ppmv during the day and 100 ppmv
during the night following our comparison (Fig. 2). The
increase of the diurnal observation error would mainly de-
crease the error reduction by few percents but won’t affect
seriously its spatial distribution. For future inversions, the
preliminary analysis of flux model errors, and simulated con-
centrations, will guide the definition of the final state vector,
especially the choice of a daily mean or diurnal and noctur-
nal components. The prior flux correlations will be guided
by analysis of flux tower measurements. The error reduc-
tion presented here would be affected by these additional el-
ements depending on the hypothesis introduced in the inverse
system. (Sarrat et al., 2007). The nocturnal accumulation of CO2 in
the boundary layer at Marmande tower remains lower than
observed in the direct simulation (Fig. 2b). The offline cou-
pling with the lagrangian model reinforces this additional
vertical diffusion. The integration of the particles near the
surface is optimal between 50 m to 100 m high during the
night, higher than expected. The reduced height of the noc-
turnal boundary layer explains the diurnal variability. The
absolute error on the boundary layer height during the night
induces a larger relative error compared to daytime, which is
directly related to the modelled CO2 concentration. The er-
ror reduction of the diurnal CO2 fluxes is less affected by the
larger nocturnal observation error than nocturnal CO2 fluxes. But the final error reduction is larger for the nocturnal com-
ponent of the fluxes. Considering the inversion setup for
one mean flux, the error reduction using the two towers of
the campaign is reduced but still covers a few tenths of km
(Fig. 9). We suppose in this experiment the model is able to
reproduce the diurnal variability of the CO2 fluxes, as shown
in the study of Ahmadov et al. (2007) during the same cam-
paign using the coupled model WRF-VPRM. 8
Conclusions Chevallier, F., Viovy, N., Reichstein, M., and Ciais, P.:
On
the assignment of prior errors in Bayesian inversions of
CO2 surface fluxes,
Geophys. Res. Letters,
33,
L13802,
doi:10.1029/2006GL026496, 2006. We have developed a demonstration system for regional in-
verse modelling at the meso scale, and tested it for the
CERES intensive campaign (2nd IOP). Using available mea-
surement locations and dates, a large part (more than 50%)
of the domain (300 km×300 km) is constrained with an er-
ror reduction larger than 30%. No spatial a priori correla-
tion was used to enlarge the impact of the data. Concentra-
tion measurements on towers play a major role in reducing
uncertainties for surface fluxes, whereas aircraft measure-
ments above the planetary boundary layer influence mostly
the boundaries. The intensive period of flights improved the
error reduction at the surface by 15 to 20% compared with
the two tower-only case. This implies also a reduced impor-
tance of boundary conditions compared to surface influence
for near ground observations. Davis, K., Bakwin, P., Berger, B., Yi, C., Zhao, C., Teclaw, R., and
Isebrands, J.: The annual cycle of CO2 and H2O exchange over
a northern mixed forest as observed from a very tall tower, Glob. Change Biol., 9, 1278–1293, 2003. Dolman, A. J., Noilhan, J., Durand, P., Sarrat, C., Brut, A., Piguet,
B., Butet, A., Jarosz, N., Brunet, Y., Loustau, D., Lamaud, E.,
Tolk, L., Miglietta, R. R. F., Gioli, B., Magliulo, V., Esposito,
M., Gerbig, C., Krner, S., Galdemard, P., Ramonet, M., Ciais,
P., Neininger, B., Hutjes, R. W. A., Macatangay, J. A. E. R.,
Schrems, O., P´erez-Landa, G., Sanz, M. J., Scholz, Y., Facon,
G., Ceschia, E., and Beziat, P.: CERES, the CarboEurope Re-
gional Experiment Strategy in Les Landes, South West France,
May–June 2005, Bull. Am. Meteorol. Soc., 87(10), 1367–1379,
doi:10.1175/BAMS–87–10–1367, 2006. Dolman, H., DeMartino, B., Gioli, B., Hutjes, R. W. A., Lin-
droth, A., Miglietta, F., Millan, M. M., Sanz, M. J., and Schu-
macher, M.: Regional assessment and monitoring of the car-
bon balance within Europe (RECAB): Experimental strategy and
mesoscale modeling preliminary results, European Geophysical
Union, Nice, EGS02-A-06856, 2002. Acknowledgements. We wish to thank all participants in the
CERES campaign for making their data freely available on the
official website (http://carboregional.mediasfrance.org/campagne/
index). CarboEurope is supported by the European Commission
under the 6 Framework Programme. 7
Discussion This can be approximated with vertical turbulent veloc-
ities of a few meters per second and mixing depths of one
to two kilometers for a strongly convective diurnal bound-
ary layer such as 27 May. The present LPDM parameteriza-
tion for boundary layer turbulence corresponds to Gaussian
but inhomogeneous conditions (Thomson, 1987; Du, 1997). Using enhanced mixing in a test, we found that error reduc-
tion was smaller due to a loss of particles to the free tro-
posphere. A more sophisticated planetary boundary layer
scheme should be developed to improve the vertical mix-
ing in LPDM. We also examined the vertical mixing over
the ocean, mostly dominated by wind shear, weaker than ob-
served at the Biscarosse tower due to the complex dynamical
processes induced by the highly negative energy balance at
the surface. This development needs the description of other Overall, for short term mesoscale inversions, the need for
constraint at the boundaries is clear for observations in the
free troposphere, but not in the boundary layer. Global mod-
els seem able to give this information at least in the context
of weak mean wind and strong convection near the surface. The use of global models to constrain boundaries might also
be critical when using tall towers. As shown by the third ex-
periment, such towers are mainly influenced by large scale
motions. The high spatial resolution used for this inversion shows
that the most observable flux contribution comes from re-
gions close to the measuring instruments within the bound-
ary layer. This spatial distribution is strongly dependent on
the quality of atmospheric transport, but mesoscale models
like MesoNH have shown their ability to reproduce complex
dynamical processes in the lower atmosphere during the day Atmos. Chem. Phys., 8, 3459–3471, 2008 www.atmos-chem-phys.net/8/3459/2008/ 3469 T. Lauvaux et al.: Mesoscale inversion CO2 fluxes, 1988–2003, Global Biogeochem. Cy., 20, GB1002,
doi:10.1029/2004GB002439, 2006. CO2 fluxes, 1988–2003, Global Biogeochem. Cy., 20, GB1002,
doi:10.1029/2004GB002439, 2006. dynamic processes such as entrainment and detrainment at
the top of the planetary boundary layer. We will investigate
such a scheme in the context of an inversion using actual
measurements. Ba Baldocchi, D. D., Falge, E., Gu, L., Olson, R., Hollinger, D.,
Running, S., Anthoni, P., Bernhofer, C., Davis, K., Fuentes, J.,
Goldstein, A., Katul, G., Law, B., Lee, X., Malhi, Y., Meyers,
T., Munger, J. W., Oechel, W., Pilegaard, K., Schmid, H. 7
Discussion P.,
Valentini, R., Verma, S., Vesala, T., Wilson, K., and Wofsy, S.:
FLUXNET: A new tool to study the temporal and spatial variabil-
ity of ecosystem-scale carbon dioxide, water vapor, and energy
flux densities, Bull. Am. Meteorol. Soc., 82, 2415–2435, 2001. Finally, towers are the most important source of informa-
tion for the inversion of surface fluxes. At the same time,
aircraft measurements allow us to constrain the limits with
only a small effect on the final surface fluxes and their pos-
terior uncertainties. The combination of these observations
defines a complete framework to assess sources and sinks for
mesoscale domains. Bousquet, P., Peylin, P., Ciais, P., Qu´er´e, C. L., Friedlingstein, P.,
and Tans, P. P.: Regional changes in carbon dioxide fluxes of
land and oceans since 1980, Science, 290, 1342–1346, 2000. Carouge, C.: Vers une estimation des flux de CO2 Europ´een, PhD
thesis, 2006. 8
Conclusions This research was funded in
part by the US Department of Commerce, National Oceanic and
Atmospheric Administration (NOAA), Office of Global Programs,
Global Carbon Cycle program, grant number NA040AR4310124. Du, S.: Universality of the lagrangian velocity structure function
constant (C0) across different kinds of turbulence, Bound. Lay. Met., 83, 207–219, 1997. Enting, I. G.: Inverse Problems in Atmospheric Constituent Trans-
port, Cambridge University Press, 2002. Edited by: I. Aben Edited by: I. Aben Enting, I. G., Trudinger, C. M., and Francey, R. J.: A synthesis in-
version of the concentration and δ13C of atmospheric CO2, Tel-
lus, 47B, 35–52, 1995. T. Lauvaux et al.: Mesoscale inversion T. Lauvaux et al.: Mesoscale inversion Peylin, P., Rayner, P. J., Bousquet, P., Carouge, C., Hourdin, F.,
Ciais, P., Heinrich, P., and AeroCarb Contributors: Daily CO2
flux estimate over Europe from continuous atmospheric mea-
surements: Part 1 inverse methodology, Atmos. Chem. Phys., 5,
3173–3186, 2005,
http://www.atmos-chem-phys.net/5/3173/2005/. ill
hi k
i
A S
i
h A Gerbig, C., Lin, J. C., Wofsy, S. C., Daube, B. C., Andrews,
A. E., Stephens, B. B., Bakwin, P. S., and Grainger, C. A.:
Toward constraining regional-scale fluxes of CO2 with atmo-
spheric observations over a continent: 1. Observed spatial vari-
ability from airborne platforms, J. Geophys. Res., 108(D24),
4756, doi:10.1029/2002JD003018, 2003. Gerbig, C., Lin, J. C., Wofsy, S. C., Daube, B. C., Andrews, A. E.,
Stephens, B. B., Bakwin, P. S., and Grainger, C. A.: Toward
constraining regional-scale fluxes of CO2 with atmospheric ob-
servations over a continent: 2. Analysis of COBRA data using a
receptor-oriented framework, J. Geophys. Res., 108(D24), 4756,
doi:10.1029/2003JD003770, 2003. Peters, W., Miller, J., B., Whitaker, J., Denning, A., S., Hirsch, A.,
Krol, M., C., Zupanski, D., Bruhwiler, L., Tans, P., P.: An en-
semble data assimilation system to estimate CO2 surface fluxes
from atmospheric trace gas observations, J. Geophys. Res., 110,
24304, doi:10.1029/2005JD006157, 2005. Raupach, M. R., Marland, G., Ciais, P., Le Quere, C., Canadell,
J. G., Klepper, G., and Field, C. B.: Global and regional drivers
of accelerating CO2 emissions, PNAS, 104, 10 288–10 293, http:
//www.pnas.org/cgi/content/abstract/104/24/10288, 2007. Gloor, M., Fan, S. M., Pacala, S., and Sarmiento, J.: Optimal sam-
pling of the atmosphere for purpose of inverse modeling: A
model study, Global Biogeochem. Cy., 14, 407–428, 2000. Gloor, M., Bakwin, P., Hurst, D., Lock, L., Draxler, R., and Tans, P.:
What is the concentration footprint of a tall tower?, J. Geophys. Res., 106, 17 831–17 840, 2001. Rayner, P. J. and O’Brien, D. M.: The utility of remotely sensed
CO2 concentration data in surface source inversions, Geophys. Res. Let., 28, 175–178, 2001. Gurney, K. R., Law, R. M., Denning, A. S., Rayner, P. J., Baker,
D., Bousquet, P., Bruhwiler, L., Chen, Y.-H., Ciais, P., Fan, S.,
Fung, I. Y., Gloor, M., Heimann, M., Higuchi, K., John, J.,
Maki, T., Maksyutov, S., Masarie, K., Peylin, P., Prather, M.,
Pak, B. T. Lauvaux et al.: Mesoscale inversion C., Randerson, J., Sarmiento, J., Taguchi, S., Takahashi,
T., and Yuen, C.-W.: Towards robust regional estimates of CO2
sources and sinks using atmospheric transport models, Nature,
415, 626–630, 2002. Rayner, P. J., Scholze, M., Knorr, W., Kaminski, T., Giering, R.,
and Widmann, H.: Two decades of terrestrial Carbon fluxes from
a Carbon Cycle Data Assimilation System (CCDAS), Glob. Bio-
geochem. Cy., 19, 2026, 2005. R¨odenbeck, C., Houweling, S., Gloor, M., and Heimann, M.: Time-
dependent atmospheric CO2 inversions based on interannually
varying tracer transport, Tellus B, 55, 488–497, 2003. Rodgers, C. D.: Inverse methods for atmospheric sounding: Theory
and practice, World Scientific Publishing Co. Pte. Ltd. 2000. Keeling, C., D., Piper, S., C., Heimann, M.: A Three-dimensional
Model of Atmospheric CO2 transport based on observed winds:
4. Mean annual gradients and interannual variations, AGU, As-
pects of Climate Variability in the Pacific and the Western Amer-
icas, Geophysical Monograph 55, 305–363, 1989. Sarrat, C., Noilhan, J., Lacarr`ere, P., Donier, S., Dolman, H.,
Gerbig, C., Ciais, P., and Butet, A.: Atmospheric CO2 mod-
eling at the regional scale:
Application to the CarboEu-
rope Regional Experiment, J. Geophys. Res., 112, D12105,
doi:10.1029/2006JD008107, 2007. Krinner, G., Viovy, N., de Noblet-Ducoudr´e, N., Ogee, J., Polcher,
J., Friedlingstein, P., Ciais, P., Sitch, S., and Prentice, I. C.:
A dynamic global vegetation model for studies of the cou-
pled atmosphere-biosphere system, Global Biogeochem. Cy., 19,
GB1015, doi:10.1029/2003GB002199, 2005. Sarrat, C., Noilhan, J., Dolman, H., Gerbig, C., Ahmadov, R., Tolk,
L. F., Meesters, A. G. C. A., Hutjes, R. W. A., Ter Maat, H. W.,
P´erez-Landa, G., and Donier, S.: Atmospheric CO2 modeling
at the regional scale: an intercomparison of 5 meso-scale atmo-
spheric models, Biogeosciences Discuss., 4, 1923–1952, 2007,
http://www.biogeosciences-discuss.net/4/1923/2007/. Lafore, J., Stein, J., Bougeault, P., Ducrocq, V., Duron, J., Fis-
cher, C., Hereil, P., Mascart, P., Masson, V., Pinty, J. P., Re-
delsperger, J., Richard, E., and de Arellano, J. V.: The Meso-NH
atmospheric simulation system. Part I: adiabatic formulation and
control simulations, Ann. Geophys., 16, 90–109, 1998,
http://www.ann-geophys.net/16/90/1998/. Seibert, P. and Frank, A.: Source-receptor matrix calculation with a
Lagrangian particle dispersion model in backward mode, Atmos. Chem. Phys., 4, 51–63, 2004 Stephens, B. B., Wofsy, S. C., Keeling, R. F., Tans, P., and Potosnak,
M. J.: The CO2 Budget and Rectification Airborne Study: Strate-
gies for measuring rectifiers and regional fluxes, Inverse Methods
in Global Biogeochemical Cycles, Geophysical Monograph 114,
AGU, 2000. Law, R. References Filippi, D., Ramonet, M., Ciais, P., Picard, D., Roulley, J.-C. L.,
Schmidt, M., and Nedelec, P.: Greenhouse airborne measure-
ments over Europe, Geophys. Res. Abstracts, 5, 2003. Ahmadov R., Gerbig, C., Kretschmer, R., Koerner, S., Neininger,
B.,
Dolman,
A. J.,
and Sarrat,
C.:
Mesoscale covari-
ance of transport and CO2
fluxes:
Evidence from ob-
servations and simulations using the WRF-VPRM coupled
atmosphere-biosphere model, J. Geophys. Res., 112, D22107,
doi:10.1029/2007JD008552, 2007 Geels, C., Gloor, M., Ciais, P., Bousquet, P., Peylin, P., Vermeulen,
A. T., Dargaville, R., Aalto, T., Brandt, J., Christensen, J., Frohn,
L. M., Haszpra, L., Karstens, U., R¨odenbeck, C., Ramonet, M.,
Carboni, G., and Santaguida, R.: Comparing atmospheric trans-
port models for future regional inversions over Europe. Part 1:
Mapping the CO2 atmospheric signals, Atmos. Chem. Phys., 7,
3461–3479, 2007, Baker, D. F., Law, R. M., Gurney, K. R., Rayner, P., Peylin, P.,
Denning, A. S., Bousquet, P., Bruhwiler, L., Chen, Y.-H., Ciais,
P., Fung, I. Y., Heimann, M., John, J., Maki, T., Maksyu-
tov, S., Masarie, K., Prather, M., Pak, B., Taguchi, S., and
Zhu, Z.: TransCom 3 inversion intercomparison: Impact of
transport model errors on the interannual variability of regional http://www.atmos-chem-phys.net/7/3461/2007/. http://www.atmos-chem-phys.net/7/3461/2007/. Atmos. Chem. Phys., 8, 3459–3471, 2008 www.atmos-chem-phys.net/8/3459/2008/ 3470 www.atmos-chem-phys.net/8/3459/2008/ Atmos. Chem. Phys., 8, 3459–3471, 2008 T. Lauvaux et al.: Mesoscale inversion M., Rayner, P. J., Steele, L. P., and Enting, I. G.:
Data and modelling requirements for CO2 inversions using
high frequency data, Tellus, 55B, 512–521, doi:10.1034/j.1600-
0560.2003.0029.x, 2003. Tans, P., Fung, I. Y., and Takahashi, T.: Observational constraints on
the global atmospheric CO2 budget, Science, 247, 1431–1438,
1990. Michalak, A. M., Hirsch, A., Bruhwiler, L., Gurney, K. R., Pe-
ters, W., and Tans, P. P.: Maximum likelihood estimation of
covariance parameters for Bayesian atmospheric trace gas sur-
face flux inversions, J. Geophys. Res., 110(D24), D24107,
doi:10.1029/2005JD005970, 2005. Tarantola, A.: Inverse Problem Theory: Methods for Data Fitting
and Parameter Estimation, Elsevier, Amsterdam, 1987. P´erez-Landa, G., Ciais, P., Gangoiti, G., Palau, J. L., Carrara, A.,
Gioli, B., Miglietta, F., Schumacher, M., Millan, M. M., and
Sanz, M. J.: Mesoscale circulations over complex terrain in the
Valencia coastal region, Spain, Part 2: linking CO2 surface fluxes
with observed concentrations, Atmos. Chem. Phys., 7, 1851–
1868, 2007,
http://www.atmos-chem-phys.net/7/1851/2007/. Thomson, D. J.: Criteria for the selection of stochastic models of
particle trajectories in turbulent flow, J. Fluid Mech., 180, 529–
556, 1987. Uliasz, M.: Lagrangian particle modeling in mesoscale applica-
tions, Environmental Modelling II, ed. P. Zanetti, Computational
Mechanics Publications, 71–102, 1994. Uliasz, M., Denning, A. S., Schuh, A., Richardson, S. J., Miles,
N., Davis, K. J., and Zupanski, D.: Estimation of regional CO2 http://www.atmos-chem-phys.net/7/1851/2007/. Atmos. Chem. Phys., 8, 3459–3471, 2008 www.atmos-chem-phys.net/8/3459/2008/ 3471 T. Lauvaux et al.: Mesoscale inversion Zupanski,
D.,
Denning,
A. S.,
Uliasz,
M.,
Zupanski,
M.,
Schuh, A. E., Rayner, P. J., Peters, W., and Corbin, K.:
Carbon flux bias estimation employing Maximum Likelihood
Ensemble Filter (MLEF), J. Geophys. Res., 112, D17107,
doi:10.1029/2006JD008371, 2007. fluxes using concentration measurements from the ring of towers
in northern Wisconsin, American Geophysical Union, Fall Meet-
ing, 2005. fluxes using concentration measurements from the ring of towers
in northern Wisconsin, American Geophysical Union, Fall Meet-
ing, 2005. Vermeulen, A. T., Eisma, R., Hensen, A., and Slanina, J.: Transport
model calculations of NW-European methane emissions, Env. Sci. and Policy, 2, 315–324, 1999. Wang, Y., P., Leuning, R., Cleugh, H., Coppin, A.: Parameter es-
timation in surface exchange models using non-linear inversion:
How many parameters can we estimate and which measurements
are most useful?, Glob. Change Biol., 7, 495–510, 2001. Atmos. Chem. Phys., 8, 3459–3471, 2008 www.atmos-chem-phys.net/8/3459/2008/
|
https://openalex.org/W1980921371
|
https://pub.uni-bielefeld.de/download/2726546/2729563/fpsyg-06-00316.polyanskaya.pdf
|
English
| null |
Perception of speech rhythm in second language: the case of rhythmically similar L1 and L2
|
Frontiers in psychology
| 2,015
|
cc-by
| 13,907
|
ORIGINAL RESEARCH
published: 25 March 2015
doi: 10.3389/fpsyg.2015.00316 Perception of speech rhythm in
second language: the case of
rhythmically similar L1 and L2
Mikhail Ordin* and Leona Polyanskaya*
Fakultät für Linguistik und Literaturwissenschaft, Universität Bielefeld, Bielefeld, Germany We investigated the perception of developmental changes in timing patterns that happen
in the course of second language (L2) acquisition, provided that the native and the target
languages of the learner are rhythmically similar (German and English). It was found that
speech rhythm in L2 English produced by German learners becomes increasingly stress-
timed as acquisition progresses. This development is captured by the tempo-normalized
rhythm measures of durational variability. Advanced learners also deliver speech at
a faster rate. However, when native speakers have to classify the timing patterns
characteristic of L2 English of German learners at different proficiency levels, they attend
to speech rate cues and ignore the differences in speech rhythm. Edited by:
Judit Gervain Edited by:
Judit Gervain,
Université Paris Descartes, France Edited by:
Judit Gervain,
Université Paris Descartes, France Reviewed by:
Juan M. Toro,
Universitat Pompeu Fabra, Spain
Pilar Prieto,
Universitat Pompeu Fabra, Spain
*Correspondence:
Mikhail Ordin and Leona Polyanskaya,
Fakultät für Linguistik und
Literaturwissenschaft, Universität
Bielefeld, Bielefeld 33615, Germany
mikhail.ordin@uni-bielefeld.de;
leona.polyanskaya@uni-bielefeld.de Specialty section: Specialty section:
This article was submitted to
Language Sciences, a section of the
journal Frontiers in Psychology Received: 08 November 2014
Accepted: 05 March 2015
Published: 25 March 2015 Keywords: speech rhythm, rhythm metrics, durational variability, rhythm acquisition, rhythm perception, timing
patterns, rhythm development, second language Keywords: speech rhythm, rhythm metrics, durational variability, rhythm acquisition, rhythm perception, timing
patterns, rhythm development, second language Introduction Reviewed by:
Juan M. Toro,
Universitat Pompeu Fabra, Spain
Pilar Prieto,
Universitat Pompeu Fabra, Spain Reviewed by:
Juan M. Toro,
Universitat Pompeu Fabra, Spain
Pilar Prieto,
Universitat Pompeu Fabra Spain The differences between languages and linguistic varieties are manifested in the acoustic compo-
nents of the signal that are perceived by the auditory system and cognitively processed to extract
linguistic structures. Some minute acoustic differences are perceived by the native speakers, while
some gross acoustic changes in the speech stream may be ignored—either not perceived not
attended to. In this study we concentrated on the perceptual relevance of the changes in speech
rhythm in second language (L2) that happen in the course of L2 acquisition, provided that the
native and the target languages of the learner are rhythmically similar. *Correspondence:
Mikhail Ordin and Leona Polyanskaya,
Fakultät für Linguistik und
Literaturwissenschaft, Universität
Bielefeld, Bielefeld 33615, Germany
mikhail.ordin@uni-bielefeld.de;
leona.polyanskaya@uni-bielefeld.de g
g
g
y
y
We start by introducing the notion of rhythm. Further we move on to discussing why perception
of rhythmic patterns might be linguistically relevant. Then we report how speech rhythm develops
in L2 English spoken by German learners, and why it is worth studying whether people are sensi-
tive to the changes in L2 speech rhythm, when the target and native languages of the learner are
rhythmically similar. A brief overview of empirical studies in German and English speech rhythm
is provided to highlight rhythmic similarities between languages. Later, we report the results of the
perception experiment aimed to answer the main question of the research: are the rhythmic changes
that happen in the course of acquisition perceptually relevant, if the L1 and L2 of the learner are
rhythmically similar? In the end, we show the theoretical implications of our findings. where Those languages that produce the
effect of stress-timing display vowel reduction, more complex C
clusters, have more different syllable types, opposition between
phonologically long and short vowels, between geminate and
non-geminate consonants, are less likely to exhibit vowel har-
mony and fixed stress. Rhythm metrics reflect these language-
specific phonological properties. To name a few examples, 1C
is thought to be indicative of the syllabic structure, syllable com-
plexity and consonantal phonotactic constraints. 1V is supposed
to be indicative of the degree of vowel reduction. VarcoV and
VarcoC reflect the same properties 1C and 1V do, but Varco
measures are supposed to neutralize the effect of the tempo dif-
ferences, and thus to reduce the effect of idiosyncrasies in speech
production. %V indicates the syllabic structure and inventory. Languages with more restricted syllabic inventory operate less
complex syllables, usually of the CV structure. The more types
of syllables there are in the language inventory, the more conso-
nants are added to the onset or coda of the syllables. This reduces
the proportion of vocalic intervals to the overall duration of the
utterance, and %V decreases. Prieto et al. (2012) demonstrated
that prosodic edges and heads are marked by manipulating dura-
tional ratios in a language-specific way, and this may also account
for small differences in rhythm measures between languages. The
analysis of the surface durational variability of V and C intervals
indeed allows spreading the languages on a continuous scale with
respect to their rhythmic properties and to say that one language
is more or less stress-timed than another, or that the two lan-
guages are rhythmically similar. However, it did not yet provide
an unambiguous support for the Rhythm Class Hypothesis that
suggests that languages are split into distinct rhythm categories. Some of these metrics are influenced by the speech rate to a
higher degree than the others (Dellwo and Wagner, 2003; Dellwo,
2006; Wiget et al., 2010). For example, 1V depends on the mean
duration of vowels in an utterance. That is, if speech is deliv-
ered at a faster rate, mean durations become smaller and 1V
tends to decrease. Dellwo (2006) suggested Varco measure to
normalize for the tempo differences and to capture the differ-
ences in durational variability irrespective of the differences in
speech rate between the languages. where where m—number of interval in an utterance for which PVI is
calculated, m—number of interval in an utterance for which PVI is
calculated, d—duration of kth interval. Higher values of %V and lower values of the other metrics cor-
respond to the languages that are traditionally defined as syllable-
timed (Ramus et al., 1999; Low et al., 2000; Grabe and Low, 2002;
Dellwo and Wagner, 2003; White and Mattys, 2007), and pos-
sibly provide the necessary cues to differentiate the durational
patterns of the rhythmically contrastive languages (Ramus and
Mehler, 1999; Ramus et al., 1999). A new concept of speech rhythm has been introduced in an
attempt to find the perceptually relevant acoustic correlates of
rhythmic patterns (Ramus et al., 1999). It rests on the assumption
that consonantal and vocalic intervals in the speech signal can
exhibit language-specific patterns of durational variability. Lan-
guages that are traditionally classified as “stress-timed” exhibit
higher degree of durational variability compared to “syllable-
timed” languages. That is, stress-timing is characterized by more
substantial differences in duration of vowels and consonantal
clusters within the same utterance produced by the same speaker. To capture the variability in duration of speech intervals, a num-
ber of the so-called rhythm metrics have been proposed. Among
the most commonly used interval-based rhythm metrics are the
pairwise variability index (PVI) (Grabe and Low, 2002), the stan-
dard deviation in duration of speech intervals (1) and the per-
centage of vocalic material in an utterance (%V) (Ramus et al.,
1999), the coefficient of variation in duration of speech inter-
vals (Varco) (Dellwo and Wagner, 2003). Conventionally these
metrics are applied to vocalic (V) and consonantal (C) inter-
vals, i.e., sequences of consecutive vowels or consonantal clus-
ters that can straddle the syllabic and word boundaries within an
utterance). Yet the metrics have also been applied to capture the
durational variability of other speech intervals in order to inves-
tigate the multiple rhythms on multiple timescale, e.g., on the
timescale of feet (Nolan and Asu, 2009) or syllables (Ordin et al.,
2011). ,
;
,
)
Dauer (1983, 1987) and Schiering (2007) analyzed the phono-
logical structure of languages which give the impression of stress-
timing or syllable-timing. where Grabe and Low (2002) also
suggested a normalized version of PVI in an attempt to neutralize
the influence of the speech rate on the measures of the local dura-
tional variability. Formulas 1 and 2 show how the raw (rPVI) and
the normalized (nPVI) versions are calculated. White and Mat-
tys (2007) and Wiget et al. (2010) reported Varco measures, %V
and nPVI are more robust to the fluctuations of the speech rate
compared to non-normalized metrics. Citation: Ordin M and Polyanskaya L (2015)
Perception of speech rhythm in
second language: the case of
rhythmically similar L1 and L2. Front. Psychol. 6:316. doi: 10.3389/fpsyg.2015.00316 The word rhythm implies the idea of periodicity. Based on the auditory impression that certain
events or certain speech constituents reoccur periodically in the speech stream, the languages
were classified into stressed-timed (in which stressed syllables were perceived to be distributed at
roughly equal intervals, e.g., German, English, Dutch, Russian) and syllable-timed (in which all
syllables were perceived to be of roughly equal duration, e.g., French, Italian, Spanish). Later, a new March 2015 | Volume 6 | Article 316 Frontiers in Psychology | www.frontiersin.org 1 Perception of speech rhythm in L2 Ordin and Polyanskaya r PVI =
"m −1
X
k = 1
dk −dk + 1
/(m −1)
#
(2) rhythmic class of mora-timed languages (in which moras are sup-
posedly perceived as roughly equal in duration, e.g., Japanese,
West Greenlandic) was added. Experimental studies, however,
failed to find empirical evidence to support this impression
(Roach, 1982; Dauer, 1983; Pamies Bertran, 1999). However,
adults (Ramus et al., 1999) and even infants (Nazzi et al., 1998;
Ramus and Mehler, 1999; Nazzi and Ramus, 2003) are able to
differentiate between rhythmic patterns of languages that are
traditionally classified as stress- and syllable-timed. Therefore,
researchers continued looking for the acoustic correlates of audi-
torily perceived differences in speech rhythm. (2) Frontiers in Psychology | www.frontiersin.org Rhythm Changes in Second Language
Acquisition Papers focussed on acquisition of speech rhythm in L2 are rare. Most of these studies concentrate on comparing rhythm in L2
speech with the target represented by an adult native speaker. Examined L2 speech is usually produced by rather advanced
learners. The results showed that the rhythm scores in L2 speech
are intermediate between those in the native and the target lan-
guage of the learners (White and Mattys, 2007). This is usually
interpreted as the influence of the native language of the learner
on his speech production in the L2. Low et al. (2000) showed that
nPVI-V in L2 Singaporean English is influenced by the L1 Chi-
nese language. Rhythm in L2 English was shown to be affected by
L1 Chinese, French, Spanish, Romanian and Italian (White and
Mattys, 2007; Gut, 2009; Mok, 2013, etc.). These important findings regarding sensitivity of listeners to
rhythmic differences have been done using discrimination tests. We know that people can discriminate between utterances even
with small differences in durational variability. However, certain
functions attributed to speech rhythm are not based on discrim-
ination, but rather on classification (segmentation, evaluation of
accentedness, detection of linguistic origin of the speaker, etc.). Classification is different from discrimination. The listener may
be able to perceive some acoustic differences when attending to
them, but nevertheless ignore these differences when attributing
an acoustic signal to a certain group, or when making a decision
whether an acoustic signal is a representative of a certain class. In this particular study we focused not merely on whether the
differences in L2 rhythm between utterances delivered by learn-
ers at different proficiency levels are detected. We were rather
interested in whether listeners are able to reliably classify the
utterances of L2 learners into distinct classes based on timing
differences between utterances, and if so, which timing patterns
listeners use to form the classes. The studies with the emphasis on development of rhythmic
patterns in the course of L2 acquisition are even rarer. One of
the few exceptions is the study by Ordin and Polyanskaya (2014)
who compared how speech rhythm develops in L1 and in L2
acquisition. They found that speech rhythm develops from more
syllable-timed toward more stress-timed patterns both in child
L1 and in adult L2 speech. Research Question Previous studies have showed that rhythmic patterns change as
language acquisition progresses even when the native and the
target languages of the learners are rhythmically similar (Ordin
et al., 2011). In our study, we were interested whether these
developmental changes in speech rhythm are perceptually rel-
evant. It is already known that the listeners are sensitive to
the rhythmic differences between rhythmically contrastive lan-
guages (Ramus and Mehler, 1999) as well as between German
and English, i.e., between rhythmically similar languages (Vicenik
and Sundara, 2013). Listeners are also able to distinguish rhyth-
mic patterns of the utterances from the same language (White
et al., 2012; Arvaniti and Rodriquez, 2013). Therefore, we think
that the fine distinctions between rhythmic patterns typical of L2
English of adult learners at different proficiency levels might be
detected. Importance of Rhythmic Patterns for Speech
Processing This led to the hypoth-
esis that alongside with constructing the first representation of
their native language, babies use rhythmic patterns to bootstrap
on the syntactic properties of the language and on lexicon
(Christophe and Dupoux, 1996; Mazuka, 1996; Mehler et al.,
1996, 2004; Nespor et al., 1996). Rhythmic patterns are also used
to develop strategies for segmentation of continuous speech
and consequent word extraction and learning (Christophe et al.,
2003; Thiessen and Saffran, 2007). In light of these considera-
tions, we could suggest that the ability to recognize the durational
cues pertaining to the speech rhythm is of the utmost importance
for language acquisition and speech processing (e.g., for devel-
opment and implementation of language-specific segmentation
strategies). Therefore, sensitivity to timing differences, which is
already observed in infancy, also persists in adulthood (Ramus
and Mehler, 1999; White et al., 2012). Adults also use rhythmic
cues to recognize the foreign accent in L2 speech and to detect
the linguistic origin of the speaker (Kolly and Dellwo, 2014), to
evaluate the degree of accentedness in L2 speech (Polyanskaya
et al., 2013), to extract discrete linguistic units from continuous
speech (Christophe et al., 2003). German share phonological parameters that are known to affect
the rhythm metrics. Both of these languages are classified as
stress-timed in terms phonetic timing patterns captured by met-
ric scores (Grabe and Low, 2002) and exhibit the phonological
characteristics typical of stress-timed languages (Dauer, 1987;
Schiering, 2007). Therefore German learners of English do not
have to acquire phonological characteristics like production of
complex syllables and complex consonantal clusters, opposition
of long and short vowels, etc. Table 1 provides the metric scores
in monolingual adult speech delivered by adult native speak-
ers of either German or English, as reported in various studies. No unambiguous tendency is evident as for in which of these
languages the durational variability is higher. %V seems a bit
lower in German, which can be explained by a slightly higher
syllabic complexity and a higher number of C clusters in Ger-
man than in English (Delattre, 1965 cited in Gut, 2009). Com-
parison of the metric scores for German and English with those
reported for traditional syllable-timed languages (Ramus et al.,
1999; Grabe and Low, 2002; White and Mattys, 2007) shows
that both German and English exhibit higher duration variability
and lower %V. Importance of Rhythmic Patterns for Speech
Processing People are sensitive to the timing patterns which are cap-
tured by rhythm metrics. Mehler et al. (1996) hypothesized
that
pre-linguistic
infants
perceive
incoming
continuous
speech as a succession of vocalic and consonantal segments,
vocalic
segments
are
processed
as
informative
harmonic
signals of variable duration and intensity, which are alter-
nating with unanalyzed noise (consonantal intervals). This n PVI = 100 ×
"m −1
X
k = 1
dk −dk + 1
(dk + dk + 1)/2
/(m −1)
# (1) March 2015 | Volume 6 | Article 316 Frontiers in Psychology | www.frontiersin.org 2 Perception of speech rhythm in L2 Ordin and Polyanskaya Time-Intensity-Grid-Representation of incoming continuous
speech is based on innate perception mechanisms that help them
to construct the first representation of their language. Ramus
et al. (1999) observed that languages with similar rhythmic
properties tend to share more typological characteristics of
grammatical and phonological structure. This led to the hypoth-
esis that alongside with constructing the first representation of
their native language, babies use rhythmic patterns to bootstrap
on the syntactic properties of the language and on lexicon
(Christophe and Dupoux, 1996; Mazuka, 1996; Mehler et al.,
1996, 2004; Nespor et al., 1996). Rhythmic patterns are also used
to develop strategies for segmentation of continuous speech
and consequent word extraction and learning (Christophe et al.,
2003; Thiessen and Saffran, 2007). In light of these considera-
tions, we could suggest that the ability to recognize the durational
cues pertaining to the speech rhythm is of the utmost importance
for language acquisition and speech processing (e.g., for devel-
opment and implementation of language-specific segmentation
strategies). Therefore, sensitivity to timing differences, which is
already observed in infancy, also persists in adulthood (Ramus
and Mehler, 1999; White et al., 2012). Adults also use rhythmic
cues to recognize the foreign accent in L2 speech and to detect
the linguistic origin of the speaker (Kolly and Dellwo, 2014), to
evaluate the degree of accentedness in L2 speech (Polyanskaya
et al., 2013), to extract discrete linguistic units from continuous
speech (Christophe et al., 2003). Time-Intensity-Grid-Representation of incoming continuous
speech is based on innate perception mechanisms that help them
to construct the first representation of their language. Ramus
et al. (1999) observed that languages with similar rhythmic
properties tend to share more typological characteristics of
grammatical and phonological structure. Frontiers in Psychology | www.frontiersin.org Participants Piske et al. (2001) analyzed a range of factors that influence pro-
nunciation of L2 learners. These factors, among others, included
the age and the length of exposure to the L2, amount of L2
use, language learning aptitude and motivation, learning mode. In our study we controlled these factors by collecting the rel-
evant information in a detailed language-background question-
naire (see Appendix 1 in online Supplementary Materials). Based
on the questionnaire, we selected only those speakers who formed
a homogeneous group and varied only in the degree of L2 mas-
tery. The relevant information gleaned from the questionnaire
was further verified in an informal interview during the recording
sessions. Speech Material Germany at the time of the recordings. However, they reported
to have little to no command of German, lived in close English-
speaking community at the UK military bases in Nord-Rhein
Westphalia, worked in only English-speaking environment, had
English as their home and neighborhood language, came from
monolingual English-speaking families and were raised in mono-
lingual environment. Rhythm Changes in Second Language
Acquisition The authors showed that both vocalic
and consonantal variability in duration in L2 English increases as
a function of the length of residence in the UK in adult speech
when the target (English) and the native (Italian or Punjabi)
languages of the learners are rhythmically contrastive. Ordin et al. (2011) showed that durational variability in
speech of L2 learners also increases with proficiency growth
when the target (English) and the native (German) languages
of the learners exhibit similar rhythmic properties. English and March 2015 | Volume 6 | Article 316 Frontiers in Psychology | www.frontiersin.org 3 Perception of speech rhythm in L2 Ordin and Polyanskaya TABLE 1 | Metric scores for German and English as reported in various studies. Language
Rhythm metrics
References
%V
VarcoV
n-PVI-V
rPVI-C
1C
VarcoC
German
41.7
52.5
68.7
65.0
Russo and Barry, 2008
42.8
71.7
Dellwo and Wagner, 2003
46.4
59.7
55.3
52.6
Grabe and Low, 2002
39.8
51.5
53.6
67.0
62.0
54.0
Arvaniti (2012)—overall score
36
44
55
73
62
51
Arvaniti (2012)—scores obtained on read sentences that were deliberately designed to
enhance durational variability
41
52
56
60
54
50
Arvaniti (2012)—scores obtained on read sentences that were deliberately designed to
inhibit durational variability
41
52
53
56
55
50
Arvaniti (2012)—scores obtained on sentences uncontrolled for phonotactics
42
55
52
72
55
50
Arvaniti (2012)—scores obtained on spontaneous speech
English
38.0
64.0
73.0
70.0
59.0
White and Mattys, 2007
42.0
55.7
Dellwo and Wagner, 2003
41.1
57.2
64.1
56.7
Grabe and Low, 2002
40.1
53.5
Ramus et al., 1999
45.7
54.8
59.9
68.9
60.0
Arvaniti (2012)—overall score
41
48
55
83
68
57
Arvaniti (2012)—scores obtained on read sentences that were deliberately designed to
enhance durational variability
50
46
51
57
49
53
Arvaniti (2012)—scores obtained on read sentences that were deliberately designed to
inhibit durational variability
44
50
56
61
55
55
Arvaniti (2012)—scores obtained on sentences uncontrolled for phonotactics
48
66
66
77
68
59
Arvaniti (2012)—scores obtained on spontaneous speech TABLE 1 | Metric scores for German and English as reported in various studies. Frontiers in Psychology | www.frontiersin.org Elicitation Procedure The selected learners of English first underwent a pronuncia-
tion test so that we could assess the learners’ mastery of pro-
nunciation. The test was devised by the authors and consisted
of two parts: Perception and production. The perception part
was compiled from Vaughan-Rees (2002) and included phoneme
recognition, emotion recognition, intention recognition tasks. The production part included sentence reading. The sentences
for production were composed to evaluate segmental realizations
and prosodic control of the participants in the second language. The test ran for approximately 20 min. The test and the details on
the controlled pronunciation features and assessment criteria can
be found in Appendix 2 in online Supplementary Materials. We have recorded 51 German learners of L2 English (17–35
years old, M = 21; 27 females). We selected for participation
only those people who grew up in or near the city of Bielefeld
in North-Rhein Westphalia. The variety of German spoken in
that region closely resembles what is understood as a Northern
standard variety of German (Hochdeutsch). The selected partic-
ipants did not exhibit features of regional varieties of German. All the participants were monolingual native speakers of German
without speech or hearing disorders. Further on, a 5-min phonetic aptitude test (PAT) was admin-
istered. The authors devised this test based on the oral mimicry
tests described by Pike (1959), Suter (1976), and Thompson
(1991). The test is aimed to predict the general phonetic ability by
asking the participant to imitate novel sounds that do not exist in
their native or target language and to mimic novel prosodic phe-
nomena (e.g., lexical tones, tonal contours with accents which are We have also recorded 10 native speakers of English (south-
ern British variety, 25–40 years, M = 30, 6 females) to compare
the metric scores of the L2 learners of English with those of
the L1 English speakers. The English speakers were residents in March 2015 | Volume 6 | Article 316 Frontiers in Psychology | www.frontiersin.org 4 Perception of speech rhythm in L2 Ordin and Polyanskaya not aligned according to the convention of the learner’s target or
native language, etc.). The test and the details on the assessment
criteria can be found in Appendix 3 in online Supplementary
Materials. The sounds to imitate were presented by the holder of
the IPA certificate confirming his proficiency in producing and
perception of sounds existing in world languages. 1According to the evaluators’ opinion, we did not have true beginners, and our
participants better correspond to the lower-intermediate (B1.1 according to the
Common European Framework for Languages), upper-intermediate (B2.1) and
advanced (C1.1 and C1.2) levels. CEFL specifies skills the learner should achieve
at each of the six levels: A1, A2, B1, B2, C1, C2. However, as each level is usu-
ally covered in language schools during two intensive courses, teachers split each
level into two sublevels, e.g., C1.1 and C1.2. However, official CEFL guidelines do
not split six levels into sublevels, and division into C1.1 and C1.2 is done—rather
arbitrarily—by the teachers. We did not want to resort to commonly used place-
ment tests to evaluate the learners’ proficiency level because standard placement
tests are designed to make an initial assessment to place the student into the course
that fits his level. Placement tests are not designed to estimate the proficiency level
for certification of the achieved proficiency level. Therefore, using several human
evaluators (to avoid human bias) is the best methodological option, in our opinion. Elicitation Procedure The perfor-
mance of participants in PAT did not correlate with their L2 pro-
ficiency (we had both high and low proficiency learners with both
high and low phonetic aptitude). Neither did the performance of
the L2 learners in the PAT correlate with their performance in the
English pronunciation test with any of the metrics calculated on
their speech. This shows that the ability to imitate rhythmic pat-
terns of the target language is not related to the general phonetic
aptitude and we can eliminate a potential alternative explanation
that the differences in rhythmic patterns between learners at dif-
ferent proficiency levels are pertaining to the phonetic aptitude
rather than to the overall proficiency. ratings between the teachers, we used Cronbach alpha, which
is 0.90 for vocabulary, 0.89 for fluency and 0.92 for grammati-
cal accuracy. This shows high agreement between the raters and
confirms the reliability of their assessments. We averaged three
ratings across the parameters for each rater and each interview,
and thus got three mean ratings per learner. The teachers’ assessments and the results of the pronunciation
tests were used to place the learner into one of the following profi-
ciency groups: beginners (12 speakers with ratings between 4 and
6), intermediate (9 speakers with mean ratings between 6 and 8),
and advanced learners (22 speakers with ratings above 8)1. We
used the results of the pronunciation test to assess the pronun-
ciation skills of the learners. Eight speakers were not attributed
to any group, either because the teachers did not agree with
each other in their assessments (2 speakers were excluded for
this reason) or because of the discrepancy between the results of
the pronunciation tests and the teachers’ assessment of accuracy,
fluency and vocabulary resources. Pronunciation skills do not
always agree with the general assessment of the learner’s reading,
writing, listening and speaking skills, vocabulary size, grammar
accuracy, etc. That is why we deemed it necessary to combine the
tutors’ assessment of fluency, accuracy and vocabulary on the one
hand and the mastery of pronunciation on the other hand. In case
when pronunciation lags far behind the general L2 mastery or
exceeds the expected level, the learner was not attributed to any
of the proficiency groups. At the next stage, an informal interview was conducted by
the first author. Segmentation Thirty three elicited sentences per speaker were annotated in
Praat (Boersma and Weenink, 2010). Annotation was performed
by the second author. Each sentence was divided into V and C
intervals. The segmentation was carried out manually by the sec-
ond author based on the criteria outlined in Peterson and Lehiste
(1960) and Stevens (2002) for V and C intervals. The burst of energy corresponding to the release of the closure
was taken as the starting point of a consonantal interval with the
initial voiceless plosive sound after a pause and at the beginning
of a sentence. Either the stop release, or apparent beginning of a
voice bar, or other cues indicating apparent vibration of the vocal
folds (whatever came first) were considered as the beginning of
a consonantal interval with the initial voiced plosive. The mark-
ers of the turbulent noise were taken as the beginning of fricative
consonants. The beginning of the first formant was taken as the
beginning of a sonorant consonant. Consonantal intervals in the The tests and recordings were made individually with every
participant in a sound-treated booth of the audio-visual studio at
the Bielefeld University in Germany. The recordings were made
in WAV PCM at 44 kHz, 16 bit, mono. Elicitation Procedure General questions about preferences in read-
ing and music, lifestyle, career choice, biography, and childhood
were asked (Appendix 4 in online Supplementary Materials). The
interviews were recorded and lasted approximately 12 min long
with each participant. Following the interview, we ran a sentence elicitation task,
similar to one used by Bunta and Ingram (2007). Thirty three
sentences were elicited from each speaker. We used 33 pic-
ture prompts for the elicitation procedure. The participants
viewed picture slides in PowerPoint presentation. Each slide was
accompanied with a descriptive sentence. The participants were
instructed to remember the sentences. The participants could
move to the next image or to go back to the previous slide at
their own pace. When they had viewed all the slides, they were
asked to look at the images again, without the accompanying text,
and to recall and say the sentences that they had been asked to
remember. In a very rare case (<5%) when the speaker could not
remember the sentence or retrieved a modified sentence from
memory, verbal prompts were used to help the speaker to pro-
duce the correct sentence. For example, the participant said “The
dog is running after the cat,” and the expected sentence was “The
dog is chasing the cat.” The researcher responded to the partici-
pant: “Yes, it is. You could also say chasing, which means running
after. Can you say what you see at this picture once again?” One
verbal prompt was sufficient to elicit the expected sentence when
there was a mismatch in the first trial. The recording ran con-
tinuously throughout the sentence elicitation procedure. The list
of elicited sentences and the examples of picture prompts can be
found in Appendix 5 in online Supplementary Materials. Assessment of Learners’ Proficiency Three experienced teachers of English as a foreign language lis-
tened to the recorded interviews and evaluated learners’ fluency,
grammatical accuracy, and vocabulary resources. They used a 10-
point scale for each parameter. To estimate the consistency of March 2015 | Volume 6 | Article 316 Frontiers in Psychology | www.frontiersin.org 5 Perception of speech rhythm in L2 Ordin and Polyanskaya middle of a sentence were considered to start after the vowel fin-
ishes, and to stretch until the onset of the following vowel. The
end of the consonantal interval in the final position was marked
at the end of the acoustic energy. The consonantal intervals in
the final positions were considered to start immediately after the
vowel and finish at the end of the fricative noise (for obstruents)
or at the end of the first formant (for sonorants). Conventional
procedure based on the analysis of the waveform and the spec-
tral characteristics of the speech signal was based to identify the
boundaries of the vocalic intervals. The end of the vowel was
identified by the abrupt change in the vowel formant structure
or by termination of the formants, and by the significant drop in
the waveform amplitude. The onset of the vowel was marked at
the beginning of the voicing identified as the start of the regular
vertical stripes on the spectrogram in the region of the second
and higher formats. The marker indicating the vowel onset or
offset was placed at the point closest to the zero crossing on the
waveform. sentence to account for possible developmental changes in speech
tempo in the course of L2 acquisition, and for the interaction of
speech rhythm and speech tempo. Although some rhythm metrics were claimed to be better
than others at quantifying rhythm, there is no consensus on
which metrics have more discriminative power. White and Mat-
tys (2007), for example, advocated for pairwise metrics, while
Ramus et al. (1999) favored 1C and %V. Loukina et al. (2011)
performed the analysis of 15 rhythm metrics and in experiments
separating pairs of languages by rhythmic properties showed that
a rhythm measure that is successful at separating one pair often
performs poorly at separating another pair. Considering the lack
of consensus on the optimal set of metrics, we decided not to limit
our investigation to the metrics which were found more useful
in certain studies. Assessment of Learners’ Proficiency Metric
Description
%V
Percentage of vocalic intervals
1V
Standard deviation of vocalic intervals duration
1C
Standard deviation of consonantal intervals duration
nPVI-V
Averaged of the mean differences between successive vocalic
intervals
nPVI-C
Averaged of the mean differences between successive consonantal
intervals
rPVI-V
Averaged difference in duration of successive vocalic intervals
rPVI-C
Averaged difference in duration of successive consonantal intervals
VarcoV
Coefficient of variation of vocalic intervals, i.e., standard deviation
divided by the mean
VarcoC
Coefficient of variation of consonantal intervals, i.e., standard
deviation divided by the mean
MeanV
Mean duration of vocalic intervals
MeanC
Mean duration of consonantal intervals TABLE 2 | Metrics used in this study. Pauses and hesitations were not included into V or C intervals
and were discarded. If the same type of the interval was annotated
prior and following the pause, we treated them as two separate
intervals because they are likely to be perceived as such. Final
syllables were included into analysis. March 2015 | Volume 6 | Article 316 Assessment of Learners’ Proficiency Instead we tested all the metrics in order to
see which ones better capture the differences in rhythm between
sentences produced by L2 learners at different proficiency levels. A series of by-sentence ANOVA tests (Table 3) with the values
of the metrics as the dependent variables and proficiency level as
the factor shows that non-normalized rhythm metrics (1V, 1C,
rPVI-v, rPVI-c) and %V do not differ between the proficiency
levels. As the raw metrics do not differ between the proficiency
levels, we are not including them into further statistical tests. In difficult cases where it was necessary to place the boundary
between the consonantal interval represented by a sonorant con-
sonant with a clear formant structure and a vowel, the decision
was based on the amplitude of the first format. Such difficult cases
were associated with the boundaries or categorizing allophones
of /l/ (e.g., in the words girl, ball, table). We based our segmen-
tation on purely phonetic criteria, therefore /l/ was sometimes
marked as a vowel (in case of a vocalized [l]), and sometimes
as a consonant. The decision was based on (1) auditory analy-
sis by an experienced phonetician, and (2) amplitude of the first
formant. If the amplitude did not drop after the preceding vowel
and the segment was perceived by a phonetician as a vocalized [l],
then the segment was segmented as a vocalic interval. We did not
want to pre-define certain types of segments either as consonan-
tal or vocalic. We adopted a phontic approach to speech rhythm. Within the adopted framework, speech rhythm is represented by
the surface timing patterns, which are purely phonetic, and pho-
netic properties are not discrete and cannot be pre-assigned to a
certain phonological category a-priory. TABLE 2 | Metrics used in this study. Calculating the Rhythm Metrics The sentences elicited using the picture prompts were used to
calculate the rhythm metrics. The sentence elicitation procedure
helped us to avoid the reading mode and made speech material
more similar to natural spontaneous speech. Besides, we obtained
lexically identical sentences from every participant, which is nec-
essary to analyze the development of speech rhythm per se, not
affected by the differences between the sentences in phonotactics,
number of syllables in polysyllabic words, syntactic structures
and phrasing. TABLE 3 | Non-significant ANOVA tests for the rhythm metrics between
proficiency groups. Metric
Significance of
Significance of welch test
levene’s test
(if Levene’s test is significant) or
F statistics of the analysis of variance
rPVI-v
0.513
0.4
rPVI-c
0.007
0.267
1V
0.748
0.692
1C
< 0.0005
0.154
%V
0.691
0.068
March 2015 | Volume 6 | Article 316 TABLE 3 | Non-significant ANOVA tests for the rhythm metrics between
proficiency groups. g
Traditional rhythm metrics were calculated on each sentence. The overview of the selected metrics was given in the Table 2. We
also calculated the mean duration of V and C intervals for each Frontiers in Psychology | www.frontiersin.org Perception of speech rhythm in L2 Ordin and Polyanskaya ANOVAs on the rate-normalized rhythm measures revealed
significant difference between proficiency levels at p < 0.0005
for each metric. These metrics were included into multivari-
ate model. The MANOVA test with nPVI metrics, Varco met-
rics and mean durations of V and C intervals as the dependent
variables and proficiency level as the factor revealed a significant
effect of proficiency level on the rhythm measures, 3 = 0.856,
F(12, 2822) = 19.06, p < 0.0005, µ2 = 0.075. Figures 1–3 show
that the metric scores increase as L2 acquisition progresses, which
indicates that German learners of English deliver L2 speech at
a higher rate and with higher degree of stress-timing as their
L2 mastery grows. The differences between the proficiency lev-
els pairwise for each metric are mostly significant (significance
values are given in Table 4). the pairwise durational variability and speech rate discriminate
between the proficiency levels much better than utterance-wise
variability. We also wanted to see how close the advanced German learn-
ers of English are to their target in regard to acquisition of rhyth-
mic patterns. Calculating the Rhythm Metrics For this, we compared the metric scores calculated
on the sentences produced by the advanced learners of English
with those calculated on the sentences spoken by native English
speakers. T-tests (Table 7) reveal that the metric scores do not FIGURE 2 | nPVI-v and nPVI-r in the sentences produced by native
English speakers and by German learners of English at beginning,
intermediate and advanced proficiency levels. Error bar shows 95%
confidence interval. The MANOVA was followed up with the discriminant anal-
ysis. We used only those metrics that were found to differ sig-
nificantly between proficiency levels in our previous tests. The
analysis revealed two discriminant functions. The first function
explained 96.9% of variance, canonical R2 = 0.14, and the sec-
ond explained only 3.1% of variance, R2 = 0.005. In combination
these functions significantly differentiated the proficiency levels,
3 = 0.856, χ2
(12) = 220.318, p < 0.005. The second function
alone did not significantly differentiate between the proficiency
levels, 3 = 0.995, χ2
(5) = 7.232, p = 0.204. This can also be
seen on the discriminant function plot (Figure 4). Classification
results (Table 5) show that the model classifies correctly 57% of
cases (chance is 33%). The correlations between the outcomes and discriminant
functions revealed that the measures of local—pairwise—
variability and of speech rate loaded on the first function, and
global measures of variability loaded more highly on the second
function (see Table 6). As the first function explains substantially
more variance that the second function, we can conclude that FIGURE 2 | nPVI-v and nPVI-r in the sentences produced by native
English speakers and by German learners of English at beginning,
intermediate and advanced proficiency levels. Error bar shows 95%
confidence interval. FIGURE 1 | VarcoV and VarcoC in the sentences produced by native
English speakers and by German learners of English at beginning,
intermediate and advanced proficiency levels. Error bar shows 95%
confidence interval. FIGURE 3 | meanV and meanC in the sentences produced by native FIGURE 3 | meanV and meanC in the sentences produced by native
English speakers and by German learners of English at beginning,
intermediate and advanced proficiency levels. Error bar shows 95%
confidence interval. FIGURE 3 | meanV and meanC in the sentences produced by native
English speakers and by German learners of English at beginning,
intermediate and advanced proficiency levels. Error bar shows 95%
confidence interval. Calculating the Rhythm Metrics FIGURE 1 | VarcoV and VarcoC in the sentences produced by native
English speakers and by German learners of English at beginning,
intermediate and advanced proficiency levels. Error bar shows 95%
confidence interval. March 2015 | Volume 6 | Article 316 Frontiers in Psychology | www.frontiersin.org 7 Perception of speech rhythm in L2 Ordin and Polyanskaya TABLE 4 | Significance for comparisons of rhythm metrics between proficiency levels pairwise (with Hochberg’s correction). Comparison
VarcoV
VarcoC
nPVI-V
nPVI-C
meanV
meanC
Beginner—Intermediate
<0.0005
0.855
<0.0005
0.17
0.004
0.315
Intermediate—Advanced
0.328
<0.0005
0.096
<0.0005
0.004
0.011
TABLE 6 | Structure matrix of the discriminant function coefficients. Function
1
2
meanV
−0.5∗∗
0.07
nPVI-C
−0.472∗∗
−0.404
nPVI-V
−0.469∗∗
−0.444
meanC
−0.347∗∗
−0.224
VarcoC
0.436
0.79∗∗
VarcoV
0.409
−0.552∗∗
The stars indicate larger correlation between each variable and one of the discriminant
functions. TABLE 4 | Significance for comparisons of rhythm metrics between proficiency levels pairwise (with Hochberg’s correction). 4 | Significance for comparisons of rhythm metrics between proficiency levels pairwise (with Hochberg’s correction). TABLE 6 | Structure matrix of the discriminant function coefficients. TABLE 6 | Structure matrix of the discriminant function coefficients. Function
1
2
meanV
−0.5∗∗
0.07
nPVI-C
−0.472∗∗
−0.404
nPVI-V
−0.469∗∗
−0.444
meanC
−0.347∗∗
−0.224
VarcoC
0.436
0.79∗∗
VarcoV
0.409
−0.552∗∗
The stars indicate larger correlation between each variable and one of the discriminant
functions. FIGURE 4 | Discriminant function plot. purposes of marking edges and heads of prosodic constituents
(see Prieto et al., 2012). FIGURE 4 | Discriminant function plot. The analysis shows that speech rate and the degree of
stress-timing increase as a function of proficiency growth. This
tendency, however, can only be captured by normalized rhythm
metrics. Raw metrics do not differ between the proficiency lev-
els. The values of the raw metrics are influenced by the speech
tempo, i.e., by the mean durations of speech intervals: The faster
one talks, the shorter the V and S intervals become; the shorter
speech intervals result in smaller durational differences in pairs
of consecutive intervals and in smaller standard deviation in
duration of speech intervals. As the mean durations of speech
intervals significantly differ between the proficiency levels, we
should also expect significant differences in the values of the
raw metrics. However, this was not confirmed. Calculating the Rhythm Metrics We believe that
the values of the raw metrics are influenced by two conflicting
forces: The tendency to deliver speech at a faster rate and with
higher durational variability at high proficiency levels. This con-
flict prevents the emergence of significant differences in the val-
ues of raw metrics between proficiency levels. Normalization—
removing the influence of speech tempo—allows us to notice the
trend to enhance durational variability in L2 speech with profi-
ciency. The lack of significant differences in %V between the pro-
ficiency levels presents an interesting case. In earlier studies %V
has been reported to be robust to fluctuations in speech tempo
(Wiget et al., 2010) and to discriminate between L1 and L2 speech
(White and Mattys, 2007). However, in our experiment %V was
not informative. %V is the proportion of the vocalic material in
a sentence, and that is determined by phonotatic differences. The
proportion of vocalic material will be lower in the languages that
allow complex consonantal clusters and reduction of vowels in
unstressed positions (e.g., German, Russian, English). These lan-
guages are traditionally classified as stress-timed (Dauer, 1983). The languages on the opposite end of the spectrum impose strong
phonotactic constraints, prefer simple CV syllables and feature TABLE 5 | Classification Results based on the Discriminant Analysis. Predicted Group Membership (in %)
Beginners
Intermediate
Advanced
Original Group
Beginners
38.4
0
55.1
Membership (in%)
Intermediate
26.6
0.3
73.1
Advanced
9.8
0
90.2 TABLE 5 | Classification Results based on the Discriminant Analysis. differ between sentences spoken by advanced German learners
and native speakers of English, with the exception of meanC
(overall shorter C intervals in the utterances of L2 speakers)
and rPVI-C (raw pairwise variability of consonantal intervals is
higher in speech of learners of English). The difference in 1C
is on the verge of significance (p = 0.069), and the scores are
again higher in sentences produced by L2 learners. Significant
and marginally significant difference in consonantal variability
is easily accounted for the differences in articulation rate of C
intervals: longer C intervals in L2 speech result in larger standard
deviations and pairwise durational differences. What is impor-
tant is that pairwise durational variability of consonantal inter-
vals per se, i.e., when the differences in speech rate are normalized,
is also significantly higher in speech of advanced L2 learners. Frontiers in Psychology | www.frontiersin.org Calculating the Rhythm Metrics In our study, we used the
same set of sentences elicited from different speakers, thus the
lexical differences that could potentially influence %V were elim-
inated. The target and the native languages of the L2 learners
were similar in terms of phonotactic and phonological proper-
ties, and the learners did not have problems with producing the
clusters of consonants in English sentences. %V captures phono-
tactic and phonological differences, but the sentences spoken by
learners at different proficiency levels in our study manifested
only phonetic differences in timing patterns, phonotactics and
phonological characteristics were the same. Therefore, it is not
surprising that %V was not found to differ between sentences
produced by L2 learners at different proficiency levels. To conclude, the analysis confirms significant differences in
rhythmic patterns between proficiency levels in L2. Rhythm mea-
sures are more consistently stress-timed at higher proficiency
levels. Raw metrics are influenced by conflicting tendencies to
deliver speech at a faster rate and with higher durational vari-
ability at higher proficiency levels, and thus do not increase with
proficiency. The developmental tendency to increase the degree
of stress-timing in L2 speech has been observed even when both
the native and the target languages of the learner are rhythmi-
cally similar. The main research question of our study was to
investigate the perceptual relevance of the rhythmic differences
between proficiency levels. Based on the literature review, we
assumed that listeners are sufficiently sensitive to the durational
variability of C and V intervals to discriminate timing patterns of
L2 utterances delivered by learners at different proficiency levels. We wanted to find out whether the detected differences in timing
patterns between proficiency levels are used to classify utterances
into discrete categories. To address this question, we set up the
perception experiment. The discriminant analysis also reveals that the advanced learn-
ers are more consistent in realization of timing patterns com-
pared to lower-proficient learners. Inspection of the discriminant
function plot (Figure 7) reveals that the variate scores for the
advanced learners are more compact, while the variate scores for
the beginners are spread more evenly along the first discrimi-
nant function. The discriminant function plot also showed that
the variate scores for different groups of acquirers overlap (see
overlapping circles on Figure 7). This means that beginners pro-
duced sentences sometimes with high degree of durational vari-
ability, and sometimes with lower degree of durational variability. Calculating the Rhythm Metrics Advanced learners constantly produced the sentences with high
degree of durational variability. In other words, the productions
of beginners varied greatly between stress-timed and syllable-
timed rhythm patterns, but productions of advanced learners
were more consistently stress-timed. Methods
Participants We have recruited 25 native English speakers to act as listeners
in the perception study (age range—21–24 years, M = 22; 13
females). Care was taken to form a socially homogeneous group
of listeners with the same language background. All participants
were students of Ulster University, monolingual English speak-
ers (see our criteria for monolinguality in the description of the
participants for Experiment 1). All listeners grew up in or around
Belfast and were speaking the same regional variety of English
(verified by a native speaker of English, phonetician and Belfast
resident). We ensured that the participants did not differ in age,
educational level, social status, language background, experience
with foreign languages, and all had equal exposure to educated
standard British English. y
We can draw the same conclusion if we look at Table 5. Rhythm and tempo measures correctly predict the speaker’s pro-
ficiency level for 57% of sentences. The overall accuracy is sig-
nificantly above chance (33%), but the accuracy for the sentences
produced by speakers on different proficiency levels varies sub-
stantially. Sentences produced by advanced speakers were clas-
sified correctly in 90.2% of cases, while sentences produced by
beginners were classified correctly only in 38.4% of cases. This
means that the half of the sentences spoken by beginners exhibit
higher degree of variability that is typical of stress-timed rhythm
in 90.2% sentences spoken by advanced learners. On the other
hand, only 9.8% of sentences spoken by advanced learners exhibit
lower durational variability overlapping with 38.4% of sentences
from beginners. The analysis of the discriminant function plot
and the classification accuracy indicates that the timing patterns Calculating the Rhythm Metrics Advanced learners overshoot with increasing durational variabil-
ity of C intervals, although they successfully acquire variability of
V intervals. This can be explained by less assimilation of conso-
nants in clusters within syllables in L2 speech (i.e., tendency to
clearly produce all the consonants in the clusters) and by incom-
plete mastery of fine modifications in prosodic timing for the March 2015 | Volume 6 | Article 316 Frontiers in Psychology | www.frontiersin.org 8 Perception of speech rhythm in L2 Ordin and Polyanskaya TABLE 7 | t-tests comparing metric scores in English speech of native English speakers and advanced L2 learner of English. meanV
meanC
%V
1V
1C
VarcoV
VarcoC
rPVI-V
rPVI-C
nPVI-V
nPVI-C
t(1054)
1.216
3.371
−1.314
−0.064
1.818
−1.433
0.131
1.014
4.506
1.412
4.506
p
= 0.224
= 0.001
= 0.189
= 0.949
= 0.069
= 0.152
= 0.896
= 0.311
< 0.0005
= 0.158
< 0.0005 less vowel shortening (e.g., French, Japanese). These factors
increase the proportion of vocalic material in speech. Therefore,
we assume that %V is a powerful predictor to discriminate
between rhythmically contrastive languages. %V can also reflect
the differences in lexical material, i.e., whether the utterances per
se differ in phonotactic characteristics. In our study, we used the
same set of sentences elicited from different speakers, thus the
lexical differences that could potentially influence %V were elim-
inated. The target and the native languages of the L2 learners
were similar in terms of phonotactic and phonological proper-
ties, and the learners did not have problems with producing the
clusters of consonants in English sentences. %V captures phono-
tactic and phonological differences, but the sentences spoken by
learners at different proficiency levels in our study manifested
only phonetic differences in timing patterns, phonotactics and
phonological characteristics were the same. Therefore, it is not
surprising that %V was not found to differ between sentences
produced by L2 learners at different proficiency levels. become more stable and consistent as a result of the acquisition
progress. less vowel shortening (e.g., French, Japanese). These factors
increase the proportion of vocalic material in speech. Therefore,
we assume that %V is a powerful predictor to discriminate
between rhythmically contrastive languages. %V can also reflect
the differences in lexical material, i.e., whether the utterances per
se differ in phonotactic characteristics. Stimuli Supposedly,
during the training session the participants formed new percep-
tion categories for further discrimination between the stimuli
from different groups. Then the testing session began. Eighteen out of thirty tree elicited sentences per speaker were
selected for stimuli preparation. Six sentences had three stressed
syllables (e.g., the ‘dog is ‘ eating the ‘bone), six sentences included
two stressed syllables (e.g., the ‘book is on the ‘table) and six sen-
tences had only one stressed syllable (e.g., it’s ‘raining outside). The selected sentences produced by the selected speakers were
listened to in order to make sure that the sentences were indeed
pronounced with the expected number of stressed syllables. The
selected sentences are marked with asterisk in Appendix 5 in
online Supplementary Materials. We selected 378 sentences in
total for the perception experiment (21 speakers ∗18 sentences). For the testing session, we prepared 270 stimuli (different
from those used in the training session, 5 speakers per proficiency
group, 18 sentences per speaker). The procedure was the same as
in the training session, but the listeners received no feedback, and
all the stimuli were played only once. We used the speech resynthesis technique (Ramus and
Mehler, 1999) to prepare the stimuli. We replaced all consonantal
intervals in the selected sentences with “s” and all vocalic intervals
with “a” and resynthesizing sentences with constant fundamen-
tal frequency in MBROLA. The durations of “s” and “a” intervals
were equal to the duration of C and V intervals in the origi-
nal sentences. This technique degraded segmental and most of
the prosodic information from the sentences. The only preserved
differences between the identical sentences spoken by learners
at different proficiency levels were the differences in durational
ratios of C and V intervals. Regardless of the recent criticism of
this technique (Arvaniti and Rodriquez, 2013), its usefulness has
been demonstrated in a number of studies (Ramus et al., 1999;
Ramus and Mehler, 1999; Vicenik and Sundara, 2013; Kolly and
Dellwo, 2014, etc.), and we found this delexicalization method to
be optimal for the purposes of our study. The duration of the experiment varied between participants
and usually exceeded 90 min. The participants could take a short
break and have a rest pause during the training session and
between the training and the testing session, but not during
the testing session. Stimuli We selected sentences elicited from seven speakers per profi-
ciency group in the first experiment to prepare the stimuli. The
selected speakers from the advanced group had the highest mean
ratings given by the evaluators (see description of the first exper-
iment, Section Procedure). The selected speakers from the begin-
ners had the lowest mean ratings from the evaluators. We also
randomly selected seven speakers from the group of intermediate
learners. March 2015 | Volume 6 | Article 316 Frontiers in Psychology | www.frontiersin.org 9 Perception of speech rhythm in L2 Ordin and Polyanskaya Eighteen out of thirty tree elicited sentences per speaker were
selected for stimuli preparation. Six sentences had three stressed
syllables (e.g., the ‘dog is ‘ eating the ‘bone), six sentences included
two stressed syllables (e.g., the ‘book is on the ‘table) and six sen-
tences had only one stressed syllable (e.g., it’s ‘raining outside). The selected sentences produced by the selected speakers were
listened to in order to make sure that the sentences were indeed
pronounced with the expected number of stressed syllables. The
selected sentences are marked with asterisk in Appendix 5 in
online Supplementary Materials. We selected 378 sentences in
total for the perception experiment (21 speakers ∗18 sentences). We used the speech resynthesis technique (Ramus and
Mehler, 1999) to prepare the stimuli. We replaced all consonantal
intervals in the selected sentences with “s” and all vocalic intervals
with “a” and resynthesizing sentences with constant fundamen-
tal frequency in MBROLA. The durations of “s” and “a” intervals
were equal to the duration of C and V intervals in the origi-
nal sentences. This technique degraded segmental and most of
the prosodic information from the sentences. The only preserved
differences between the identical sentences spoken by learners
at different proficiency levels were the differences in durational
ratios of C and V intervals. Regardless of the recent criticism of
this technique (Arvaniti and Rodriquez, 2013), its usefulness has
been demonstrated in a number of studies (Ramus et al., 1999;
Ramus and Mehler, 1999; Vicenik and Sundara, 2013; Kolly and
Dellwo, 2014, etc.), and we found this delexicalization method to
be optimal for the purposes of our study. was played. When all 108 stimuli were presented, the partici-
pant had a 2-min break before the stimuli were played again. The training procedure was repeated three times. Results We calculated rhythm metrics on the stimuli that were classified
by the majority of listeners as Burabah, Losto, and Mahutu. The
metrics were calculated on V and C intervals. We performed the
discriminant analysis to test whether rhythm metrics statistically
discriminate between the stimuli classified into three groups. The
analysis revealed two discriminant functions. The first function
explained 94.8% of variance, R2 = 0.52, and the second function
explains 5.2% of variance, R2 = 0.05. These functions in combi-
nation significantly differentiate between the groups, λ = 0.457,
χ2
(20) = 149.9, p < 0.0005. The second function alone is not sig-
nificant, λ = 0.945, χ2
(9) = 11.101, p = 0.282. The overall accu-
racy of the model is 69% (chance is 33.3%), accuracy of Burabah
is 91%, Losto—52%, Mahutu—58.5% (chance level is 33.3% for
each category). See Table 8 for the details on the classification
accuracy. Stimuli During the experiment the participants were
offered hot and cold drinks and sweet snacks to help them cope
with possible fatigue. The participants could have their drinks
and snacks during the rest pauses as well as during the train-
ing session. The order of stimuli presentation was randomized
using the internal Praat algorithm in attempt to counterbalance
for possible fatigue effect. March 2015 | Volume 6 | Article 316 Procedure The experiment was carried out with each participant individ-
ually in the phonetic laboratory of Ulster University. The stim-
uli were presented to the listeners in two sessions: Training and
testing. The listeners were not informed that the stimuli were
derived from L2 English speech because we did not want the
listeners use linguistic expectations regarding what the stim-
uli in L2 English might sound like. This might have created
a bias that would be difficult to control. Instead, the listeners
were told that the stimuli were derived from three rare exotic
African languages. We coined these languages Burabah (sen-
tences of the advanced L2 learners converted into “sasasa” stim-
uli), Losto (stimuli based on durations in sentences of intermedi-
ate learners of English), and Mahutu (resynthesized sentences of
beginners). The structure matrix (Table 9) reveals that the first function
is loaded with the raw metrics and mean durations of V and C
intervals, while the second function is loaded with the normalized
metrics. This means that the normalized metrics cannot discrim-
inate between the groups, but mean durations and raw metrics
discriminate between the stimuli identified as Burabah, Losto,
and Mahutu with probability significantly above chance. g
We chose 108 stimuli for the training session (18 stimuli per
speaker, 2 speakers per proficiency group). Before the session,
each listener was exposed to nine stimuli, randomly selected from
those used later in the training session, 3 stimuli per proficiency
group, i.e., per “exotic language.” The listener had 1 min to listen
to these stimuli by clicking with a mouse on nine buttons on the
computer screen. Each button had a caption with the “language”
name. After 1-min familiarization, the stimuli were presented to
listener one by one. The listener had to identify from which lan-
guage (Mahutu, Losto, or Burabah) it originates. The listener was
expected to click one of the three buttons on the computer screen
with a mouse pointer. Each button had a caption with the “lan-
guage” name. On response, the listener was provided with the
feedback which “language” it really was, and the next stimulus TABLE 8 | Classification Results (prior probabilities: all groups equal). Predicted Group membership
Mahutu(%)
Losto(%)
Burabah(%)
Original
Mahutu
58.5
26.4
15.1
Losto
28.4
52.2
19.4
Burabah
1.3
7.6
91.1
March 2015 | Volume 6 | Article 316 TABLE 8 | Classification Results (prior probabilities: all groups equal). Frontiers in Psychology | www.frontiersin.org Procedure Frontiers in Psychology | www.frontiersin.org 10 Perception of speech rhythm in L2 Ordin and Polyanskaya FIGURE 6 | 1V and 1C for the stimuli identified as Mahutu, Losto, or
Burabah. Error bar shows 95% confidence interval. FIGURE 7 | meanV and meanC for the stimuli identified as Mahutu,
Losto, or Burabah. Error bar shows 95% confidence interval. durational variability for the listeners performing the classifi TABLE 9 | Structure matrix of the discriminant function coefficients. Metrics
Function
I
II
meanV
0.719∗
0.404
meanC
0.607∗
−0.181
rPVI-v
0.458∗
0.351
rPVI-c
0.371∗
−0.097
1C
0.352∗
−0.064
1V
0.421
0.559
VarcoV
−0.002
0.341∗
nPVI-c
0.102
0.208∗
nPVI-v
0.002
0.203∗
VarcoC
0.035
0.127∗
The stars indicate larger correlation between each variable and one of the discriminant
functions. TABLE 9 | Structure matrix of the discriminant function coefficients. FIGURE 6 | 1V and 1C for the stimuli identified as Mahutu, Losto, or
Burabah. Error bar shows 95% confidence interval. FIGURE 6 | 1V and 1C for the stimuli identified as Mahutu, Losto, or
Burabah. Error bar shows 95% confidence interval. FIGURE 5 | rPVI-V and rPVI-C for the stimuli identified as Mahutu,
Losto, or Burabah. Error bar shows 95% confidence interval. FIGURE 7 | meanV and meanC for the stimuli identified as Mahutu,
Losto, or Burabah. Error bar shows 95% confidence interval. FIGURE 7 | meanV and meanC for the stimuli identified as Mahutu,
Losto, or Burabah. Error bar shows 95% confidence interval. FIGURE 5 | rPVI-V and rPVI-C for the stimuli identified as Mahutu,
Losto, or Burabah. Error bar shows 95% confidence interval. FIGURE 7 | meanV and meanC for the stimuli identified as Mahutu,
Losto, or Burabah. Error bar shows 95% confidence interval. Figures 5–7 show the differences in the rhythm metrics that
significantly differ between stimuli classified into three groups. Only mean durations and non-normalized metrics (rPVI and the
standard deviation) differ significantly between the stimuli iden-
tifyed as Burabah, Mahutu, and Losto and statistically discrimi-
nate between the groups. Rhythm metrics normalized for tempo
and %V do not differ between the stimuli classified into three dif-
ferent groups, and do not discriminate between stimuli attributed
to different classes. durational variability for the listeners performing the classifi-
cation task. However, we still do not know the relative con-
tribution of the rhythm compared to tempo in classification. Procedure Step
Metrics
β
T
B
p
R2
R2 change
Significance of R2 change
1
meanV
0.525
10.1
61.108
<0.0005
0.276
0.276
<0.0005
2
meanV
0.377
7.163
43.814
<0.0005
0.386
0.110
<0.0005
meanC
0.363
6.914
46.28
<0.0005 TABLE 11 | Coefficients and parameters of the regression model with Frequency_Mahutu as the dependent variable. Step
Metrics
β
T
B
p
R2
R2 change
Significance of R2 change
1
meanV
0.525
10.1
61.108
<0.0005
0.276
0.276
<0.0005
2
meanV
0.377
7.163
43.814
<0.0005
0.386
0.110
<0.0005
meanC
0.363
6.914
46.28
<0.0005 Coefficients and parameters of the regression model with Frequency_Mahutu as the dependent variable. tempo equals 5.62 syl/s. for the stimuli identified as Burabah,
4.41 syl/s. for the stimuli identified as Losto, and 4.4 syl/s. for the
stimuli identified as Mahutu. ANOVA analysis showed that the
difference in tempo between the groups is significant, F(2, 196) =
64.077, p < 0.0005. Pairwise comparisons (with the Bonfer-
roni correction) reveal that the difference lies between “Losto”
and “Burabah” stimuli, while the difference between “Losto”
and “Mahutu” groups is not significant. Speech tempo in the
stimuli identified as Burabah is 25.7% higher than in the stim-
uli identified as Losto. This increase is above the threshold for
just noticeable tempo difference (Quene, 2007; Thomas, 2007). Speech tempo in the stimuli classified as Mahutu is 6.6% slower
than in the stimuli identified as Losto, and this difference is below
the just noticeable threshold. added, and the model was significantly improved. Adding raw
rhythm metrics as predictors did not improve the model fur-
ther. Table 10 summarized the main details of the regression
model. The results show that the most important predictors are mean
durations of V and C intervals, which are negatively correlated
with the frequency of “Burabah” response. This means that the
shorter the speech intervals (i.e., the faster the tempo), the more
likely the listener will classify the stimulus as Burabah. We also performed stepwise multiple regressions with Fre-
quency_Mahutu and with Frequency_Losto as dependent variable
(details of the regression models are in Tables 11, 12 respec-
tively). The analyses show that the most influential predictors for
both Frequency_Mahutu and Frequency_Losto are meanV and
meanC. The predictors are positively correlated with the fre-
quency of “Losto” and “Mahutu” responses, which means that the
stimuli with longer C and V intervals (i.e., slower speech rate) are
more likely to be identified as Losto or Mahutu. Procedure Listeners’ sensitivity to speech tempo can be explained by a
number of studies in physiology of hearing. Schreiner and Urbas
(1986, 1988) showed that auditory neurons fire in response to a
sharp increase in intensity that usually coincides with the vowel
onset. Consequently, the rate at which “s” and “a” alternate in the
stimuli determines the rate at which the neurons fire. Moreover,
some studies suggest a direct relation between a syllable-length
unit (“sa” unit in our stimuli) and the neural response in the
auditory cortex (Viemeister, 1988; Greenberg, 1997; Wong and
Schreiner, 2003; Greenberg and Ainsworth, 2004). Besides, the
auditory system imposes certain limitations on the speech tempo. If the assumptions to the speech rate and to the length of the
syllable-like units are violated, speech processing and decoding
of speech at the cortical level is compromised (Ghitza and Green-
berg, 2009; Ghitza, 2011). Therefore, there is a physiological basis
for discriminating fast and slow stimuli, or stimuli with longer
and shorter syllable-like units. Frontiers in Psychology | www.frontiersin.org Procedure To address this issue, we calculated the frequency for Burabah,
Losto, and Mahutu response for each stimulus, i.e., how many
listeners out of 25 identified each stimulus as Burabah (Fre-
quency_Burabah), Losto (Frequency_Losto), or Mahutu (Fre-
quency_Burabah). After that we performed stepwise multiple
regression to assess the ability of meanV, meanC, rPVI-C, rPVI-
V, 1V, and 1C to predict Frequency_Burabah. Stepwise regres-
sion was chosen because we wanted to evaluate whether both
the mean durations and the variability measures were necessary
to predict Frequency_Burabah. The constructed model included
only two steps. At the first step, meanV was entered into equation
as the most powerful predictor. At the second step, meanC was Figures 5–7 show that the stimuli identified as Burabah
exhibit shorter mean durations and smaller standard deviations
in duration of speech intervals and smaller durational differ-
ences in pairs of consecutive intervals. As the raw metrics are
influenced by the speech rate, we cannot say what the partici-
pants were listening for to make their judgments—speech tempo,
durational variability, or both. As meanV and meanC display
the highest correlations in the structure matrix (Table 9), we
could conclude that tempo is probably more important than Figures 5–7 show that the stimuli identified as Burabah
exhibit shorter mean durations and smaller standard deviations
in duration of speech intervals and smaller durational differ-
ences in pairs of consecutive intervals. As the raw metrics are
influenced by the speech rate, we cannot say what the partici-
pants were listening for to make their judgments—speech tempo,
durational variability, or both. As meanV and meanC display
the highest correlations in the structure matrix (Table 9), we
could conclude that tempo is probably more important than March 2015 | Volume 6 | Article 316 Frontiers in Psychology | www.frontiersin.org 11 Perception of speech rhythm in L2 Ordin and Polyanskaya TABLE 10 | Coefficients and parameters of the regression model with Frequency_Burabah as the dependent variable. parameters of the regression model with Frequency_Burabah as the dependent variable. 1
meanV
−0.638
−13.574
−102.93
<0.0005
0.407
0.407
<0.0005
2
meanV
−0.489
−10.506
−78.832
<0.0005
0.519
0.111
<0.0005
meanC
−0.365
−7.853
−64.49
<0.0005
TABLE 11 | Coefficients and parameters of the regression model with Frequency_Mahutu as the dependent variable. Conclusion differently by listeners with different native languages for pur-
poses of speech processing (Christophe et al., 2003; Murty et al.,
2007; Thiessen and Saffran, 2007; Kim et al., 2008). However,
their importance in processing non-linguistic stimuli when cog-
nitive mechanisms are less intensely employed might be low. When the presented stimuli are not processed as speech-like,
rhythmic differences between stimuli are not used for discrim-
ination or classification (Ramus et al., 2000). Thus, in our
experiment listeners rely more on those patterns in acoustic
signal that have direct physiological correlates rather than on
the patterns that are processed though the relay of cognitive
filters. We have shown significant differences in speech timing between
the sentences produced by the German learners of English at
different proficiency levels. As L2 acquisition progresses, L2
English is delivered at a faster rate and with a higher degree of
stress-timing. Further analysis revealed that realization of tim-
ing is more stable in L2 speech produced by advanced L2 learn-
ers. Advanced learners tend to speak consistently with a higher
degree of stress-timing. Lower proficiency speakers randomly
vary the degree of durational variability in their speech, some-
times delivering L2 speech with high durational variability, and
sometimes with a more syllable-timed rhythm. We suggest that
timing control in L2 speech production improves as acquisition
progresses, and rhythm becomes more stable. We would like to emphasize that our results do not indicate
the inability of the participants to hear the rhythmic differences. To test the ability to detect the differences in L2 speech rhythm
between proficiency levels, discrimination test is to be carried
out, and a number of studies showed that such small differ-
ences are detected. Using classification task, we can determine
which timing patterns are used to classify the utterances into
groups. It is possible, that larger differences in durational vari-
ability (e.g., between rhythmically contrastive languages that are
traditionally defined as stress-timed and syllable-timed) might
become more linguistically relevant and used in classification. Smaller differences as those revealed between L2 varieties are
not sufficiently different to be processed as linguistically rele-
vant, and timing patterns are classified based on direct phys-
iological correlates. Further research is necessary to address
which rhythmic differences could be processed as linguistically
relevant. Although rhythmic changes in L2 acquisition can be easily
profiled with normalized rhythm metrics, raw metrics do not
exhibit a clear uni-directional development. Discussion The results show that listeners classify the stimuli based on speech
tempo and ignore the differences in the durational variability
between the “sasasa” sequences. The Figures 5–7 also show that
there is no difference between the stimuli identified as Losto
and Mahutu for 1V, 1C, rPVI-V, rPVI-C, meanC, and meanV
measures. Faster stimuli with both low and high variation in
duration of V and C intervals were classified as Burabah, and
slower stimuli were almost randomly attributed to either Losto
or Mahutu. We conclude that the listeners formed only two cate-
gories: one for faster stimuli that were classified as Burabah, and
the other for slower stimuli that were randomly identified either
as Mahutu or Losto. We are not aware of any evidence of direct physiologi-
cal correlates for the ability to differentiate fine distinctions in
durational variability. Thus, we assume that differentiation of
fine distinctions in rhythmic patterns involves cognitive pro-
cessing. Peculiarities of predominant rhythmic patterns in a
certain language correlate with grammatical, morphological and
other structural characteristics. Rhythmic patterns guide the
way the language is acquired. They influence the strategies of
segmentation of continuous speech. Rhythmic cues are exploited This result agrees with psychoacoustic data in tempo percep-
tion. Quene (2007) and Thomas (2007) studied just-noticeable
differences in tempo and found that 5–8% change in tempo
(expressed as beats per minute for non-speech stimuli and
syllables-per-minute for speech stimuli) is easily detected by the
subjects. We analyzed the tempo differences between the stimuli
which were classified as Losto, Mahutu, and Burabah. Average March 2015 | Volume 6 | Article 316 Frontiers in Psychology | www.frontiersin.org 12 Perception of speech rhythm in L2 Ordin and Polyanskaya TABLE 12 | Coefficients and parameters of the regression model with Frequency_Losto as the dependent variable. Step
Metrics
β
t
B
p
R2
R2 change
Significance of R2 change
1
meanV
0.427
7.741
41.821
<0.0005
0.183
0.183
<0.0005
2
meanV
0.358
5.993
35.018
<0.0005
0.207
0.024
=0.005
meanC
0.170
2.847
18.207
=0.005
FIGURE 8 | Splitting the “sasasa” stimuli into two categories based on durational variability of vocalic and consonantal intervals and speech rate. TABLE 12 | Coefficients and parameters of the regression model with Frequency_Losto as the dependent variable. References Gut, U. (2009). Non-native Speech. A Corpus-Based Analysis of the Phonetic and
Phonological Properties of L2 English and L2 German. Frankfurt: Peter Lang. Arvaniti, A. (2012). The usefulness of metrics in the quantification of speech
rhythm. J. Phonet. 40, 351–373. doi: 10.1016/j.wocn.2012.02.003 Kim, J., Davis, C., and Cutler, A. (2008). Perceptual tests of rhythmic similarity: II. Syllable rhythm. Langu. Speech 51, 343–359. doi: 10.1177/0023830908099069 Kolly, M.-J., and Dellwo, V. (2014). Cues to linguistic origin: the contribution
of speech temporal information in foreign accent recognition. J. Phonet. 42,
12–23. doi: 10.1016/j.wocn.2013.11.004 Arvaniti, A., and Rodriquez, T. (2013). The role of rhythm class, speaking rate and
F0 in language discrimination. Lab. Phonol. 4, 7–38. doi: 10.1515/lp-2013-0002 Boersma, P., and Weenink, D. (2010). Praat: Doing Phonetics by Com-
puter (Version 5.1.22). Retrieved: December 15, 2010. Available online at:
http://www.praat.org/ Loukina, A., Kochanski, G., Rosner, B., Shih, C., and Keane, E. (2011). Rhythm
measures and dimensions of durational variation in speech. J. Acoust. Soc. Am. 129, 3258–3270. doi: 10.1121/1.3559709 Bunta, F., and Ingram, D. (2007). The acquisition of speech rhythm by bilingual
Spanish- and English-speaking four-and five-year-old children. J. Speech Lang. Hear. Res. 50, 999–1014. doi: 10.1044/1092-4388(2007/070) Low, L., Grabe, E., and Nolan, F. (2000). Quantitative characterizations of speech
rhythm: syllable-timing in Singapore English. Langu. Speech 43, 377–401. doi:
10.1177/00238309000430040301 Christophe, A., and Dupoux, E. (1996). Bootstrapping lexical acquisition: the role
of prosodic structure. Linguist. Rev. 13, 383–412. doi: 10.1515/tlir.1996.13.3-
4.383 Mazuka, R. (1996). “How can a grammatical parameter be set before the first
word?,” in Signal to Syntax: Bootstrapping from Speech to Grammar in Early
Acquisition, eds J. L. Morgan and K. Demuth (Mahwah, NJ: Lawrence Erlbaum
Associates Inc), 313–330. Christophe, A., Gout, A., Peperkamp, S., and Morgan, J. L. (2003). Discovering
words in the continuous speech stream: the role of prosody. J. Phonet. 31,
585–598. doi: 10.1016/S0095-4470(03)00040-8 Mehler, J., Dupoux, E., Nazzi, T., and Dehaene-Lambertz, G. (1996). ˇDCoping with
linguistic diversity: the infant’s viewpoint,” in From Signal to Syntax: Bootstrap-
ping from Speech to Grammar in Early Acquisition, eds J. Morgan and K. D. Demuth (Hillsdale, NJ: Erlbaum), 101–116. Dauer, R. (1983). Stress-timing and syllable-timing reanalyzed. J. Phonet. 11,
51–62. Dauer, R. (1987). “Phonetic and phonological components of language rhythm,”
in Proceedings of the 11th International Congress of Phonetic Sciences, (Tallinn,
Estonia), 447–450. Mehler, J., Sebastian-Galles, N., and Nespor, M. (2004). Conclusion Faster speech rate
at higher proficiency levels lowers the values of raw metrics,
while the need to enhance durational variability pushes the met-
rics up. These conflicting forces did not allow raw metrics to
reveal a clear developmental change as a function of acquisition
progress. Perception experiment was set up to investigate whether
monolingual English use the differences in L2 speech timing
between proficiency levels to group the utterances with differ-
ent timing patterns into the same class. Although L2 speech
indeed becomes increasingly more stress-timed with proficiency,
native speakers of English, when asked to classify different timing March 2015 | Volume 6 | Article 316 Frontiers in Psychology | www.frontiersin.org 13 Perception of speech rhythm in L2 Ordin and Polyanskaya between the stimuli are not sufficiently large to be linguistically
relevant. patterns into separate groups, paid attention to the differences
in speech rate and ignored the differences in speech rhythm
between the utterances produced by the L2 learners at differ-
ent proficiency levels. Faster utterances were grouped separately
from slower utterances. Both groups included utterances with
high and low durational variability of speech intervals. This trend
is schematically illustrated on Figure 8. The sensitivity of the
listeners to speech tempo is physiologically determined. The fact
that listeners ignore rhythmic differences in classification can
be explained by non-linguistic nature of the stimuli. Process-
ing of “sasasa” stimuli in our experiment, assumingly, does not
involve cognitive mechanisms that are employed in processing of
linguistic material, and listeners pay attention to those features
of the acoustic signal that have direct physiological correlates. Further research is necessary to understand whether the cogni-
tive filter is not applied to processing these stimuli because they
are not perceived as speech, or because the differences in rhythm References “Biological foundations
of language: language acquisition, cues for parameter setting and the bilingual
infant,” in The New Cognitive Neuroscience, ed M. Gazzaniga (Cambridge, MA:
MIT Press), 825–836. Dellwo, V. (2006). “Rhythm and speech rate: a variation coefficient for deltaC,” in
Language and Language-Processing, eds P. Karnowski and I. Szigeti (Frankfurt
am Main: Peter Lang), 231–241. Mok, P. (2013). Speech rhythm of monolingual and bilingual children at 2;06: Can-
tonese and English. Bilingualism: Language and Cognition 16, 693–703. doi:
10.1017/S1366728910000453 Dellwo, V., and Wagner, P. (2003). “Relations between language rhythm and
speech rate,” in Proceedings of the 15th International Congress of Phonetics
Sciences (Barcelona), 471–474. Murty, L., Otake, T., and Cutler, A. (2007). Perceptual tests of rhythmic similarity:
I. Mora Rhythm. Langu. Speech 50, 77–99. doi: 10.1177/00238309070500010401 Ghitza, O. (2011). Linking speech perception and neurophysiology: speech decod-
ing guided by cascaded oscillators locked to the input rhythm. Front. Psychol. 2:130. doi: 10.3389/fpsyg.2011.00130 Nazzi, T., Bertoncini, J., and Mehler, J. (1998). Language discrimination by new-
borns: toward an understanding of the role of rhythm. J. Exp. Psychol. Hum. Percept. Perform. 24, 756–766. doi: 10.1037/0096-1523.24.3.756 Ghitza, O., and Greenberg, S. (2009). On the possible role of brain rhythms in
speech perception: intelligibility of time-compressed speech with periodic and
aperiodic insertions of silence. Phonetica 66, 113–126 doi: 10.1159/000208934 Nazzi, T., and Ramus, F. (2003). Perception and acquisition of linguistic rhythm
by infants. Speech Commun. 41, 233–243. doi: 10.1016/S0167-6393(02)00106-1 Grabe, E., and Low, L. (2002). “Durational variability in speech and the rhythm
class hypothesis,” in Papers in Laboratory Phonology, Vol. 7, eds C. Gussenhoven
and N. Warner (New York, NY: Mouton de Gruyter), 515–546. Nespor, M., Guasti, M., and Christophe, A. (1996). “Selecting word order: the
rhythmic activation principle,” in Interfaces in Phonology, ed U. Kleinhenz
(Berlin: Akademie Verlag), 1–26. Greenberg, S. (1997). “Auditory function,” in Encyclopedia of Acoustics, ed M. Crocker (New York, NY: Wiley), 1301–1323. Nolan, F., and Asu, E. (2009). The pairwise variability index and coexisting
rhythms in language. Phonetica 66, 64–77. doi: 10.1159/000208931 Ordin, M., and Polyanskaya, L. (2014). Development of timing patterns in first and
second languages. System 42, 244–257. doi: 10.1016/j.system.2013.12.004 Greenberg, S., and Ainsworth, W. (2004). “Speech processing in the auditory sys-
tem: An Overview,” in Speech Processing in the Auditory System, eds S. Green-
berg, W. Ainsworth, A. Popper, and R. Fay (New York, NY: Springer-Verlag),
1–62. Ordin, M., Polyanskaya, L., and Ulbrich, C. (2011). Supplementary Material The Supplementary Material for this article can be found
online
at:
http://www.frontiersin.org/journal/10.3389/fpsyg. 2015.00316/abstract Acknowledgments We acknowledge the financial support of the German Research
Foundation (DFG) and the Open Access Publication Fund of
Bielefeld University for the article processing charge. This work
was supported by the Alexander von Humboldt foundation. We
are also thankful to Ferenc Bunta and David Ingram for sharing
the picture prompts that we used for the sentence elicitation task. References “Acquisition of timing patterns
in second language,” in Proceedings of Interspeech 2011 (Florence), 1129–1132. March 2015 | Volume 6 | Article 316 Frontiers in Psychology | www.frontiersin.org 14 Perception of speech rhythm in L2 Ordin and Polyanskaya Pamies Bertran, A. (1999). Prosodic typology: on the dichotomy between stress-
timed and syllable-timed languages. Lang. Design 2, 103–130. Suter, R. W. (1976). Predictors of pronunciation accuracy in second lan-
guage learning. Lang. Learn. 26, 233–253. doi: 10.1111/j.1467-1770.1976.tb
00275.x Peterson, G., and Lehiste, I. (1960). Duration of syllable nuclei in English. J. Acoust. Soc. Am. 32, 693–703. doi: 10.1121/1.1908183 Thiessen, E., and Saffran, J. (2007). Learning to learn: infants’ acquisition of stress-
based strategies for word segmentation. Lang. Learn. Dev. 3, 73–100. doi:
10.1080/15475440709337001 Pike, E. V. (1959). A test for predicting phonetic ability. Lang. Learn. 9, 35–41. doi:
10.1111/j.1467-1770.1959.tb01127.x Piske, T., McKay, I., and Flege, J. (2001). Factors affecting degree of foreign accent
in an L2: a review. J. Phonet. 29, 191–215. doi: 10.1006/jpho.2001.0134 Thomas, K. (2007). Just noticeable differences and tempo change. J. Sci. Psychol. 14–20. Polyanskaya, L., Ordin, M., and Ulbrich, C. (2013). “Contribution of timing pat-
terns into perceived foreign accent,” in Elektronische Sprachsignalverarbeitung
2013, ed P. Wagner (Dresden: TUDpress), 71–79. Thompson, I. (1991). Foreign accents revisited: the English pronunciation
of Russian immigrants. Lang. Learn. 41, 177–204. doi: 10.1111/j.1467-
1770.1991.tb00683.x Prieto, P., del Mar Vanrell, M., Astruc, L., Payne, E., and Post, B. (2012). Phonotac-
tic and phrasal properties of speech rhythm. Evidence from Catalan, English,
and Spanish. Speech Commun. 54, 681–702. doi: 10.1016/j.specom.2011.12.001 Vaughan-Rees, M. (2002). Test Your Pronunciation: Book With Audio CD. London:
Longman. Vicenik, C., and Sundara, M. (2013). The role of intonation
in lan-
guage and dialect discrimination by adults. J. Phonet. 41, 297–306. doi:
10.1016/j.wocn.2013.03.003 Quene, H. (2007). On the just noticeable difference for tempo in speech. J. Phonet. 35, 353–362. doi: 10.1016/j.wocn.2006.09.001 Viemeister, N. (1988). “Psychophysical aspects of auditory intensity coding,” in
Auditory Function, eds G. Edelman, W. Gall, and W. Cowan (New York, NY:
Wiley), 213–241. Ramus, F., Hauser, M., Miller, C., Morris, D., and Mehler, J. (2000). Language dis-
crimination by human newborns and by cotton-top tamarin monkeys. Science
288, 349–351 doi: 10.1126/science.288.5464.349 Ramus, F., and Mehler, J. (1999). Language identification with suprasegmental
cues: a study based on speech resynthesis. J. Acoust. Soc. Am. 105, 512–521. doi: 10.1121/1.424522 White, L., and Mattys, S. (2007). References Calibrating rhythm: first language and sec-
ond language studies. J. Phonet. 35, 501–522. doi: 10.1016/j.wocn.2x007. 02.003 White, L., Mattys, S., and Wigit, L. (2012). Language categorization by adults is
based on sensitivity to durational cues, not rhythmic class. J. Mem. Lang. 66,
665–679. doi: 10.1016/j.jml.2011.12.010 Ramus, F., Nespor, M., and Mehler, J. (1999). Correlates of linguistic rhythm in the
speech signal. Cognition 73, 265–292. doi: 10.1016/S0010-0277(99)00058-X Roach, P. (1982). “On the distinction between ‘stress-timed’ and ‘syllable-timed’
languages,” in Linguistic Controversies, ed D. Crystal (London: Edward Arnold),
73–79. Wiget, L., White, L., Schuppler, B., Grenon, I., Rauch, O., and Mattys, S. (2010). How stable are acoustic metrics of contrastive speech rhythm? J. Acoust. Soc. Am. 127, 1559–1569. doi: 10.1121/1.3293004 Russo, R., and Barry, W. J. (2008). “Isochrony reconsidered. Objectifying relations
between rhythm mesures and speech tempo,” in Proceedings of Speech Prosody
2008 (Campinas, Brazil), 419–422. Wong, S., and Schreiner, C. (2003). Representation of stop-consonants in cat pri-
mary auditory cortex: intensity dependence. Speech Commun. 41, 93–106. doi:
10.1016/S0167-6393(02)00096-1 Schiering, R. (2007). The phonological basis of linguistic rhythm. Cross-linguistic
data and diachronic interpretation. Sprachtypologie Universalienforschung 60,
337–359. doi: 10.1524/stuf.2007.60.4.337 Conflict of Interest Statement: The authors declare that the research was con-
ducted in the absence of any commercial or financial relationships that could be
construed as a potential conflict of interest. Conflict of Interest Statement: The authors declare that the research was con-
ducted in the absence of any commercial or financial relationships that could be
construed as a potential conflict of interest. Schreiner, C., and Urbas, J. (1986). Representation of amplitude modulation in the
auditory cortex of the cat. I. The anterior auditory field (AAF). Hear. Res. 21,
227–241. doi: 10.1016/0378-5955(86)90221-2 Schreiner, C., and Urbas, J. (1988). Representation of amplitude modulation in the
auditory cortex of the cat. II. Comparison between cortical fields. Hear. Res. 32,
49–64. doi: 10.1016/0378-5955(88)90146-3 Copyright © 2015 Ordin and Polyanskaya. This is an open-access article distributed
under the terms of the Creative Commons Attribution License (CC BY). The use,
distribution or reproduction in other forums is permitted, provided the original
author(s) or licensor are credited and that the original publication in this jour-
nal is cited, in accordance with accepted academic practice. No use, distribution or
reproduction is permitted which does not comply with these terms. Stevens, K. (2002). Toward a model for lexical access based on acoustic land-
marks and distinctive features. J. Acoust. Soc. Frontiers in Psychology | www.frontiersin.org References Am. 111, 1872–1891. doi:
10.1121/1.1458026 March 2015 | Volume 6 | Article 316 Frontiers in Psychology | www.frontiersin.org 15
|
https://openalex.org/W2981822968
|
https://res.mdpi.com/d_attachment/applsci/applsci-09-04524/article_deploy/applsci-09-04524-v2.pdf
|
English
| null |
15N Natural Abundance, Nitrogen and Carbon Pools in Soil-Sorghum System Amended with Natural and NH4+-Enriched Zeolitites
|
Applied sciences
| 2,019
|
cc-by
| 13,576
|
Received: 1 October 2019; Accepted: 22 October 2019; Published: 25 October 2019 Abstract: The use of rocks containing high amounts of natural zeolites (zeolitites) as soil amendment
has been found as a valuable method for increasing agriculture sustainability. However, the
potentialities and the effects of zeolitites on the biogeochemical cycles of nitrogen (N) and carbon (C)
have still not been clearly addressed in the literature. The objective of this study was therefore
to investigate the N and C pools and 15N distribution in an agricultural soil amended with both
natural and NH4+-enriched zeolitites with the aim of understanding their effects on the soil-plant
system, during sorghum cultivation, under fertilization reductions. Zeolitites were applied to an
agricultural soil both at natural state (5 and 15 kg m−2) and in an enriched state with NH4+ ions from
pig slurry (7 kg m−2). Both zeolitites at natural and enriched state increased soil cation exchange
capacity and affected microbial biomass, causing an initial decrease of microbial C and N and then a
possible increase of fungal population. N-NO3−content was lower in natural zeolitite treatments,
that lead to a lower NO3−availability for denitrifying bacteria. Zeolitites slightly affected the fixed
N-NH4+ pool. δ15N turnover indicated that N from NH4+-enriched zeolitites remained in the soil
until the growing season and that fertilizers partially substituted the fixed pool. Leaf δ15N content
indicated that plants assimilated N from NH4+-enriched zeolitites and evidenced a higher fertilization
recovery in natural zeolitite treatments. Organic C tended to be higher in all zeolitite treatment
rhizospheres. In soils amended with zeolitites at natural state (at both application rates) sorghum
yield was similar (+3.7%) to that obtained in the control while it was higher (+13.9%) in the plot
amended with NH4+-enriched zeolitites. Keywords: Zeolitite; carbon and nitrogen pools; 15N natural abundance; zeolite agriculture; microbial
biomass; nitrification and denitrification reduction applied
sciences applied
sciences applied
sciences Article
15N Natural Abundance, Nitrogen and Carbon Pools
in Soil-Sorghum System Amended with Natural and
NH4+-Enriched Zeolitites Giacomo Ferretti 1
, Barbara Faccini 1, Livia Vittori Antisari 2
, Dario Di Giuseppe 3,*
and Massimo Coltorti 1 1
Department of Physics and Earth Sciences, University of Ferrara, 44121 Ferrara, Italy;
giacomo.ferretti@unife.it (G.F.); barbara.faccini@unife.it (B.F.); massimo.coltorti@unife.it (M.C.) 1
Department of Physics and Earth Sciences, University of Ferrara, 44121 Ferrara, Italy;
giacomo.ferretti@unife.it (G.F.); barbara.faccini@unife.it (B.F.); massimo.coltorti@unife.it (M.C.)
2
Department of Agricultural and Food Science, Alma Mater Studiorum, University of Bologna,
40127 Bologna, Italy; livia.vittori@unibo.it giacomo.ferretti@unife.it (G.F.); barbara.faccini@unife.it (B.F.); massimo.coltorti@unife.it (M.C.)
2
Department of Agricultural and Food Science, Alma Mater Studiorum, University of Bologna,
40127 Bologna, Italy; livia.vittori@unibo.it 3
Department of Chemical and Geological Sciences, University of Modena and Reggio Emilia,
41121 Modena, Italy 3
Department of Chemical and Geological Sciences, University of Modena and Reggio Emilia,
41121 Modena, Italy *
Correspondence: dario.digiuseppe@unimore.it; Tel.: +39-05-9205-8497 *
Correspondence: dario.digiuseppe@unimore.it; Tel.: +39-05-9205-8497 1. Introduction The intensification of agricultural and zootechnical technology has brought food production to a
high cost in terms of environmental quality [1–4]. The large use of fertilizers, as well as the need to apply
manure, increase nutrients leaching and runoffrisk to ground and surface water (e.g., eutrophication),
soil desertification, biodiversity reduction and to an increase in greenhouse gases production (e.g., NH3,
N2O, CO2) [4,5]. www.mdpi.com/journal/applsci www.mdpi.com/journal/applsci Appl. Sci. 2019, 9, 4524; doi:10.3390/app9214524 Appl. Sci. 2019, 9, 4524 2 of 19 In this respect, natural zeolitites (NZ), particularly those containing clinoptilolite and chabazite,
have been widely used in agriculture and horticulture. Zeolitites are rocks containing more than 50% of
zeolites [6], hydrated minerals capable of binding NH4+ from solutions in their extra-framework sites,
preventing NH4+ leaching by rainfalls and/or irrigation. In this way, zeolitites can increase nitrogen use
efficiency (NUE) and behave as slow-releasing fertilizers, resulting in a more sustainable agriculture,
where the amount of fertilizers can be decreased without lowering the production yield [6–12]. Due to
their physic-chemical characteristics, zeolitites can be used as soil conditioners, carriers of fertilizers,
antibacterial agents, insecticides, pesticides, as enhancer of soil biological activity and additional
reducer of the Sodium Adsorption Ratio (SAR) index [13–19]. A yield increases of up to 65% in
several crops (sorghum, wheat, corn, rice, marrow, chard, basil, tomato, radish, watermelon, lettuce,
sunflower) in soils amended with natural or NH4+-enriched zeolitite (NEZ) were reported by several
studies [6,20,21]. Analogously, Reháková et al. [14] found an increase of roots biomass in parsley
(Petroselinum satilvum), carrots (Daucus carota), and onions (Allium cepa). The same authors detected
an increase in strawberry (Senga sengana) yield and green biomass of 70% and 49%; respectively,
when natural zeolitite was applied as fertilizer. The aforementioned studies provide very important
and useful information regarding the beneficial effects obtained using NZ in terms of crop yield,
environmental protection (reduction of nitrate leaching) and water use efficiency. Notwithstanding
these promising results, a detailed study on the N and C pools dynamics in a zeolitite-amended field is
still lacking. The implementation of the Water Framework Directive (2000/60/EC) and the Nitrate Directive
(91/976/EEC) led to the designation of large areas vulnerable to nitrate pollution (Nitrate Vulnerable Zone,
NVZ) such as the Ferrara Province (Emilia Romagna, Italy). It lays at the end of the Po Plain where the
intensive agricultural practices of the entire Emilia-Romagna are conveyed. 2.1. Zeolitites
g (
o obtain bloc The natural K-rich, Na-poor zeolitite used in this study is a byproduct from a quarry located
near Sorano village (Italy, Grosseto province) (Figure 1, 42◦41′20.65” N; 11◦44′26.29” E) that is mainly
exploited to obtain blocks and bricks for construction and gardening. It is a low-cost, natural and
eco-friendly granular by-product of the quarrying activity, part of a large zeolitized pyroclastic deposit
whose total zeolitic content is on average 70% (chabazite, 68.5%; Phillipsite, 1.8%; Analcime, 0.6%; [10]). The total CEC of the zeolitite is 2170 mmol kg−1, of which 1460 are due to Ca2+, 600 to K+, 70 to Na+
and 40 to Mg2+. The dry bulk density and water retention vary with grain size from 870 kg m−3 and
48.4% for Ø <3 mm to 560 kg m−3 and 34.2% for Ø 3–6 mm, respectively. g
g
anular by-product of the quarrying activity, part of a large zeolitized pyroclas
l zeolitic content is on average 70% (chabazite, 68.5%; Phillipsite, 1.8%; Analc
otal CEC of the zeolitite is 2170 mmol kg−1, of which 1460 are due to Ca2+, 600 t
to Mg2+. The dry bulk density and water retention vary with grain size from
for Ø <3 mm to 560 kg m−3 and 34.2% for Ø 3–6 mm, respectively. rain size distribution of the employed zeolitites are reported in Figure 2. Part g
p
y
The grain size distribution of the employed zeolitites are reported in Figure 2. Part of the NZ was
subjected to an enrichment process, which allowed the saturation of the zeolitite with NH4+, creating
the NEZ. The enrichment process was carried out by static mode in a prototype tank where the zeolitite
is mechanically mixed with pig slurry (average NH4+ content of 2000 mg L−1) at specific solid/liquid
ratio, stirring and resting times, gaining between 400 and 800 mmol of NH4+ per kg of material [23]. N and C pools of this material are reported in Table 1. ted to an enrichment process, which allowed the saturation of the zeolitite w
e NEZ. The enrichment process was carried out by static mode in a prototype ta
e is mechanically mixed with pig slurry (average NH4+ content of 2000 mg L−1)
d ratio, stirring and resting times, gaining between 400 and 800 mmol of NH4+
3]. N and C pools of this material are reported in Table 1. 1. Introduction The high number of breeding
farms located on this territory and the consolidate agricultural practice of animal slurry spreading (liquid
mixture of urine, water and feces), increases N2O emission by nitrification/denitrification and NO3−
leaching, further emphasizing the vulnerability of this province [22]. In this framework, ZeoLIFE project (LIFE10+ ENV/IT/00321) was conceived to test different
zeolitite treatments (ZT) at the field scale aiming at reducing the input of N from fertilizers and the
irrigation water in agriculture. This project exploits the high cation exchange capacity (CEC) of this
material by up-taking NH4+ from pig slurry using an especially designed prototype tank [23] or,
alternatively, from chemical fertilizers directly in the field after addition to the soil [24]. The current scientific literature lacks field studies in which a detailed investigation of the dynamics
of N and C pools in soil modified with natural zeolitites was conducted. In this paper, we present
a detailed study on the dynamics, of both organic and inorganic N and C pools in soils amended
with NZ and NEZ (e.g., exchangeable and fixed NH4+ in clay interlayers, nitrate, microbial biomass
N and C, total organic carbon and carbon in humic substances). Conventional agricultural practices (plowing, application of chemical fertilizers and monocropping)
were compared with those related to the innovative zeolitite methods. Non-amended soils (control) were
compared with both NZ and NEZ amended soils (applied at different rates), over a cultivation cycle of
sorghum (Sorghum vulgare Pers.). The main objectives of this work were therefore the following: (1)
Evaluate how the addition of NZ and NEZ affects N and C pool dynamics in the soil-plant system
over one agronomic year; (1)
Evaluate how the addition of NZ and NEZ affects N and C pool dynamics in the soil-plant system
over one agronomic year; (2)
Determine if the use of NZ or NEZ can allow a reduction in the application of chemical fertilizers
to soil; (2)
Determine if the use of NZ or NEZ can allow a reduction in the application of chemical fertilizers
to soil; (3)
Determine if the use of NZ or NEZ have beneficial effects on crop yield. Considering the global need of improving agricultural sustainability and reducing environmental
pollution, this work may help to unveil the potential of these methodologies. Considering the global need of improving agricultural sustainability and reducing environmental
pollution, this work may help to unveil the potential of these methodologies. 1. Introduction 3 of 19 Appl. Sci. 2019, 9, 4524
9, 9, x FOR PEER RE 2.1. Zeolitites
g (
o obtain bloc The average grain size of the upper soil horizon is constituted by
0.6 ± 0.1% of medium sand (200–630 µm), 7.4 ± 0.3% of fine sand (63–200 µm), 49.2 ± 3.1 of silt (2–63
µm) and 42.0 ± 3.4% of clay (< 2 µm) [28]. The average organic matter content is 8.1 ± 1.5%, hydraulic
conductivity of the soil is 1.7 ± 2.4 m day-1 while the average bulk density is 1.15 ± 0.05 kg m−3 [28]. The upper soil horizon is also generally well drained due to the presence of a sub-surface drainage
system at −1 m.b.g.l. Soil mineralogical composition is characterized by quartz, feldspar, calcite and
clay minerals (illite smectite clorite serpentine and mixed layer) [10]
The area belongs to the eastern Po River plain, where ancient interdistributary bays and brackish
marshes were recently (1860–1960) reclaimed [25,26]. The soil of the experimental field belongs to the
cartographic unit FOR1-LCO1 of the Emilia Romagna 1:50,000 Soil Map (www.regione.emiliaromagna.it),
and consisting of recent interfluvial silty-clay deposits classifiable as Calcaric Gleyic Cambisol [26,27]. The average grain size of the upper soil horizon is constituted by 0.6 ± 0.1% of medium sand
(200–630 µm), 7.4 ± 0.3% of fine sand (63–200 µm), 49.2 ± 3.1 of silt (2–63 µm) and 42.0 ± 3.4% of clay
(<2 µm) [28]. The average organic matter content is 8.1 ± 1.5%, hydraulic conductivity of the soil is
1.7 ± 2.4 m day−1 while the average bulk density is 1.15 ± 0.05 kg m−3 [28]. The upper soil horizon is
also generally well drained due to the presence of a sub-surface drainage system at −1 m.b.g.l. Soil
mineralogical composition is characterized by quartz, feldspar, calcite and clay minerals (illite, smectite,
clorite, serpentine and mixed-layer) [10]. clay minerals (illite, smectite, clorite, serpentine and mixed-layer) [10]. The experimental field was parceled in order to compare the different zeolitite treatments with
the traditional practices. The parcels were designed linear and continuous in order to facilitate the
movements of farm machines. In each parcel, three different sub-parcels were considered to consider
inter parcel variability and to have statistical significance. One parcel (0.5 ha) was amended with 7
kg m-2 (7CZ) of NEZ (Ø < 3mm); two parcels (1 ha each) were amended with 5 (5NZ) and 15 (15NZ)
kg m-2 of coarse-grained (Ø 3–6mm) NZ, respectively, and 3.5 ha were used as control (CNTR). 2.1. Zeolitites
g (
o obtain bloc 1 Location of the exploited zeolitite quarry and ZeoLIFE experimen
Figure 1. Location of the exploited zeolitite quarry and ZeoLIFE experimental field. L
ti
f th
l it d
litit
d Z
LIFE
i
Figure 1. Location of the exploited zeolitite quarry and ZeoLIFE experimental field. 4 of 19
4 of 19 Appl. Sci. 2019, 9, 4524
Appl Sci 2019 9 x FOR Figure 2. Particle size analysis of the employed zeolitites. Figure 2. Particle size analysis of the employed zeolitites. Figure 2. Particle size analysis of the employed zeolitites
Figure 2. Particle size analysis of the employed zeolitites. 2 2 Experimental Field Setting
2.2. Experimental Field Setting 2 2 Experimental Field Setting
2.2. Experimental Field Setting p
g
The experimental field is located near Codigoro town (Italy), 40 km eastward of Ferrara (Figure
1, 44°50’33’’ N and 12°05’45’’ E) and 15 km from the Adriatic Sea in a reclaimed area at an average
altitude of 3 m below sea level. The average daytime temperatures ranges from 3°C in January and
25 °C in July and the marine thermoregulation generally maintains the minima over zero, reducing
the number of night frosts. The average rainfall is between 500 and 700 mm per year, representing
the regional pluviometric minimum with peaks in autumn and summer (sub-continental climate)
The experimental field is located near Codigoro town (Italy), 40 km eastward of Ferrara (Figure 1,
44◦50’33” N and 12◦05’45” E) and 15 km from the Adriatic Sea in a reclaimed area at an average altitude
of 3 m below sea level. The average daytime temperatures ranges from 3◦C in January and 25 ◦C in
July and the marine thermoregulation generally maintains the minima over zero, reducing the number
of night frosts. The average rainfall is between 500 and 700 mm per year, representing the regional
pluviometric minimum with peaks in autumn and summer (sub-continental climate). the regional pluviometric minimum with peaks in autumn and summer (sub-continental climate). The area belongs to the eastern Po River plain, where ancient interdistributary bays and brackish
marshes were recently (1860–1960) reclaimed [25,26]. The soil of the experimental field belongs to the
cartographic
unit
FOR1-LCO1
of
the
Emilia
Romagna
1:50,000
Soil
Map
(www.regione.emiliaromagna.it), and consisting of recent interfluvial silty-clay deposits classifiable
as Calcaric Gleyic Cambisol [26,27]. 2.3. Soil and Plant Sampling A total of 60 soil samples (from 0 to 30 cm depth) were collected in the three sub-parcels for each
treatment by manual drilling using an Ejielkamp Agrisearch auger. Soil samplings were carried out in
four periods of the agronomical year: (1) November 2012 (Nov-2012), after the zeolitite application;
(2) May 2013 (May-2013), before sowing and the first fertilization; (3) June 2013 (Jun-2013), after the
urea addition; (4) September 2013 (Sep-2013), at the harvest. Soil samples collected at the harvest were
subdivided in bulk soil and rhizosphere. The rhizosphere was separated by gently shaking the plant
roots. Three representative plants from each treatment and each sub-parcel were sampled during the
harvest; each plant was subdivided in roots, stems, leaves and panicles that were subsequently dried
at 50 ◦C and milled (total of 48 samples). 2.1. Zeolitites
g (
o obtain bloc According to ZeoLIFE plan, in the first year of field experiment, sorghum (Sorghum vulgare
Pers.) was sowed on May, 9th, 2013. Nitrogen fertilizers were distributed in two steps: di-ammonium
phosphate during the sowing, with an application of 21.6 kg N ha−1 in all parcels and urea on June 3rd,
2013, with different doses in the various parcels. In the CNTR parcel, an average of 170 (±16) kg N ha−1
were supplied, while a reduction of approximately 30% and 50% of urea in NZ (both 5NZ and 15NZ
for 122 and 127 kg N ha−1, respectively) and NEZ parcels (81 kg N ha−1) was applied, respectively. Harvest occurred in September, 23rd, and the yield was separately evaluated for each treatment. 2.1. Zeolitites
g (
o obtain bloc The
smaller size of 7CZ parcel was due to the long time required to produce the NEZ using the “ZeoLIFE”
prototype tank [23,24], which has a limited production rate of about 250 kg day-1. Addition and
spreading into the field of NZ and NEZ were carried out between October 7th and November 6th,
The experimental field was parceled in order to compare the different zeolitite treatments with
the traditional practices. The parcels were designed linear and continuous in order to facilitate
the movements of farm machines. In each parcel, three different sub-parcels were considered to
consider inter parcel variability and to have statistical significance. One parcel (0.5 ha) was amended
with 7 kg m−2 (7CZ) of NEZ (Ø < 3mm); two parcels (1 ha each) were amended with 5 (5NZ) and
15 (15NZ) kg m−2 of coarse-grained (Ø 3–6mm) NZ, respectively, and 3.5 ha were used as control
(CNTR). The smaller size of 7CZ parcel was due to the long time required to produce the NEZ using
the “ZeoLIFE” prototype tank [23,24], which has a limited production rate of about 250 kg day−1. Addition and spreading into the field of NZ and NEZ were carried out between October 7th and 5 of 19 Appl. Sci. 2019, 9, 4524 November 6th, 2012. NEZ supplied approximately 410 kg N ha−1 in the 7CZ parcel. Immediately
after spreading, the field was ploughed and the zeolitite was located until an approximate depth of
30–40 cm. According to ZeoLIFE plan, in the first year of field experiment, sorghum (Sorghum vulgare
Pers.) was sowed on May, 9th, 2013. Nitrogen fertilizers were distributed in two steps: di-ammonium
phosphate during the sowing, with an application of 21.6 kg N ha−1 in all parcels and urea on June 3rd,
2013, with different doses in the various parcels. In the CNTR parcel, an average of 170 (±16) kg N ha−1
were supplied, while a reduction of approximately 30% and 50% of urea in NZ (both 5NZ and 15NZ
for 122 and 127 kg N ha−1, respectively) and NEZ parcels (81 kg N ha−1) was applied, respectively. Harvest occurred in September, 23rd, and the yield was separately evaluated for each treatment. November 6th, 2012. NEZ supplied approximately 410 kg N ha−1 in the 7CZ parcel. Immediately
after spreading, the field was ploughed and the zeolitite was located until an approximate depth of
30–40 cm. 2.4. Analytical Techniques 2019, 9, 4524 6 of 19 distilled water was added and a heating cycle of 10 min in a microwave oven was applied. Then, 20 mL
of Milli-Q H2O was added and samples were left still for 24 hours. Finally, samples were centrifuged
at 5000 rpm for 10 min and washed 2 times with 0.5 M KCl and analyzed by elemental analysis for the
evaluation of TN and δ15N [32]. Total extractable carbon (TEC) was obtained adding 100 mL of 0.1 M NaOH + 0.1 M NaP2O7·10H2O
to 10 g of air-dried soil. The solution was left in a Dubnoffbath at 60◦C for 24 hours and then centrifuged
at 6600 rpm for 15 min and filtered at 0.45 mm (Millipore). Part of this solution was acidified with
HCl 1:1 to reach pH < 2 to allow the precipitation of Humic Acids (HA), which were separated after
centrifugation at 7000 rpm for 25 min by the Non-Humic-Substances (NHS) and Fulvic Acids (FA). The
separation of FA from NHS was performed by solid chromatography using a polyvinylpyrrolidone
(PVP) polymer in acid ambient, which allows the retention of FA on the PVP and lets NHS pass through
the column. The FA was then recovered by bringing the pH of the column to alkaline values with 0.5 M
NaOH. The evaluation of C contained in TEC, HA and FA was obtained by reverse titration with 0.2 N
Fe2SO4 after oxidation of the C contained in the sample to CO2 adding 5 mL of 1 N K2Cr2O7 + 20 mL
of H2SO4 and heating at 150 ◦C for 10 min [33]. The C extraction was carried out on one replicate. g
p
Humification Index (HI) and Humification Degree (HD) were calculated as the ratio of C contained
into NHS divided by the C contained into Humic Substances (HS) and as the ratio between HS and
TEC expressed as percentage respectively (Equations (2) and (3), [33,34]). HI = [TEC −(HA + FA)]
HA + FA
(2)
HD = [100 × (HA + FA)]
TEC
(3) (2) (3) where HI is the Humification Index, DH is the Humification Degree, TEC is Total Extractable C, HA is
Humic Acids and FA are Fulvic Acids. C and N immobilized by microbial biomass (Cmic and Nmic) were obtained by the chloroform
fumigation-extraction method [35]. 2.4. Analytical Techniques CEC and exchangeable bases were determined using the Co(NH3)6Cl3 method [29,30] on both
bulk soil and rhizosphere samples collected in Sep-2013, pH was determined on 1:5 (w:v) extracts using
a pH-meter, while carbonate content was determined using the volumetric method. Total nitrogen
(TN) and organic C (TOC) of soil and plant samples were determined by an elemental CHNS-O
EA 1110 Thermo Fisher Scientific analyzer coupled with mass spectrometry (Delta Plus, Finnegan,
Thermo-Fisher). Soil samples were weighted in silver pots and treated with HCl to eliminate the
inorganic C, while plant samples were weighted in tin pots. 15N natural abundance was measured in
total and fixed nitrogen pools (TN and Fix N-NH4+) and in sorghum organs; the natural abundance was
expressed in δ% (Equation (1); [31]), where δ represents the difference from the standard atmospheric
N2 (0.3663% 15N): δ15N (%) =
15N
14N
sample −
15N
14N
st
15N
14N
st
× 1000
(1) (1) To evaluate the influence of the N source on the plant N isotopic signature, mixing lines were
built between each end-member represented by soil, NEZ and urea. To evaluate the influence of the N source on the plant N isotopic signature, mixing lines were
built between each end-member represented by soil, NEZ and urea. NO3−was extracted with Milli-Q (Millipore USA) water in a 1:5 (w/v) ratio, the solution was
shaken for 1 hour and then filtered. NO3−was determined by ion chromatography using an isocratic
dual pump ion chromatography ICS-1000 Dionex. An AS-40 Dionex auto sampler was employed to
run the analysis; Quality Control (QC) samples were run every 10 samples and the standard deviation
for all QC samples was better than 4%. Exchangeable N-NH4+ was extracted with 1M KCl in a 1:10 (w/v) ratio, the solution was shaken
for 1 hour and then filtered. The solution was diluted and analyzed with an Ion Selective Electrode
(ISE) Orion 95-12 connected to a Thermo Fisher Orion 4star pH-ISE benchtop. Interferences with Cl−
and K+ were verified and excluded by comparing measurement made with NH4Cl, CH3COONH4 +
1M KCl and CH3COONH4 + H2O standard solutions. After the elimination of organic and exchangeable N, fixed N-NH4+ (Fix N-NH4+) was determined
by adding 20 mL of KBrO to 2 g of air-dried soil samples in two replicates. After 2 hours, 40 mL of Appl. Sci. 2.4. Analytical Techniques Following this, 10 g of soil samples in four replicates were
humidified until 60% of water holding capacity (WHC) and incubated at room temperature for 1 week
at constant WHC. After this period, two replicates were extracted with 0.5 M K2SO4, representing the
non-fumigate sample (NF). On the other two samples, CHCl3 was added in a closed drier equipped
with a void pump to allow CHCl3 volatilization at room temperature. In these fumigate (F) samples
the presence of CHCl3 atmosphere guarantees the death of the microbial biomass and allows release
of the immobilized C and N after the same extraction with 0.5 M K2SO4 performed for NF. After the
extraction, both NF and F samples were analyzed with a Shimadzu Total Organic Carbon Analyser
TOC-V CPN coupled with a TN unit (TNM-1). Sorghum harvest was carried out with a harvester and
the total yield of each parcel was weighted separately directly in the farm using the local vehicle scale. It was not possible to harvest separately the three sub parcels of each treatment, as this practice would
have required an excessive time and precision not compatible with the host farm agricultural practices. 3. Results
able 1). Mo After the enrichment process with pig slurry, NEZ gains a total N load of 5.84 (± 0.90) g kg−1
(Table 1). Most of the N was present in the form of Exch N-NH4+, while Fix N-NH4+ is considerably
lower. N-NO3−load is on average 148 mg kg−1 (the material is not washed after the treatment before
the introduction into the soil system). NEZ is characterized by a very high Nmic and a low Cmic,
resulting in a Cmic/Nmic ratio lower than 1. TOC content is quite low (10.3 g kg−1 on average) resulting
in a low C/N. Soil pH was not affected by the addition of both NZ and NEZ (p > 0.05), remaining close
to neutral values in all treatments (with exception of Jun-2013 where after urea addition an increase
toward sub-alkaline values (p < 0.05), Figure 3). CEC increased significantly (p < 0.05) in the bulk
soil of all ZT. Ca2+ and Mg2+ were the major exchangeable bases, followed by K+ and Na+, with the
latter always higher in 7CZ than in the other treatments (Table 2). No significant differences were
found between the various treatments regarding exchangeable K+ (p > 0.05), while Ca2+ and Mg2+ are
generally higher in ZT, especially in the bulk soil with respect to the rhizosphere (p < 0.05). wer. N-NO3− load is on average 148 mg kg−1 (the material is not washed after the treatment b
e introduction into the soil system). NEZ is characterized by a very high Nmic and a low
sulting in a Cmic/Nmic ratio lower than 1. TOC content is quite low (10.3 g kg−1 on average) resu
a low C/N. Soil pH was not affected by the addition of both NZ and NEZ (p>0.05), remaining
neutral values in all treatments (with exception of Jun-2013 where after urea addition an inc
ward sub-alkaline values (p < 0.05), Figure 3). CEC increased significantly (p < 0.05) in the bulk
all ZT. Ca2+ and Mg2+ were the major exchangeable bases, followed by K+ and Na+, with the
ways higher in 7CZ than in the other treatments (Table 2). No significant differences were f
etween the various treatments regarding exchangeable K+ (p > 0.05), while Ca2+ and Mg
enerally higher in ZT, especially in the bulk soil with respect to the rhizosphere (p<0.05). Table 1 Average N and C pools of NH4+ enriched zeolitite (NEZ) Standard deviation within Table 1. 2.5. Statistical Analysis At least three replicates per treatments have been analyzed at each sampling point to have
statistical significance. For evaluating significant differences between the treatments, parametric
statistic was applied to the dataset. Two-Way ANOVA was employed (after verifying normality of
data distribution with Shapiro-Wilk test) for testing significant differences within factors time and
treatments and for testing interactions between factors (treatments*time). Successively, a series of
One-Way ANOVA and Tukey (HSD) post-hoc pairwise multiple comparison tests were applied at each
sapling time at “p” level of 0.05 using Sigmaplot 12.0. Correlation analyses were also performed (using
Pearson coefficient “ρ”) in order to quantify linear associations between variables for each treatment. Appl. Sci. 2019, 9, 4524
ppl. Sci. 2019, 9, x FOR P 7 of 19 7 of 19 3.1. Nitrogen and Carbon Pools in Soil Soil N pools significantly differed (p < 0.05) between the treatments and among the sampling
times, with the exception of TN that shows significant differences among sampling times only (p < 0.05)
(Table S1, Supplementary Materials). High TN were detected in Nov-2012 and Sep-2013, while a
depletion was observed in May-2013 and Jun-2013. Similar behavior is recorded for Fix N-NH4+ pool but in this case, all ZT shows slightly lower
values (p < 0.05) in Nov-2012 with respect to the CNTR, while in Jun-2013 only 15NZ treatment is
significantly lower (Table S1, Supplementary Materials). At the beginning of the experiment, N-NO3−
content was significantly higher in the CNTR and 7CZ parcels than in those amended with NZ (5NZ
and 15NZ, respectively) (p < 0.05). In May-2013 a general depletion of N-NO3−pool is noticed in all the
treatments, increasing in Jun-2013 after urea addition especially in CNTR parcel. N-NO3−values of ZT
(especially 5NZ and 15NZ) were remarkably lower in this period with respect to the CNTR (p < 0.05)
(Table S1, Supplementary Materials). Exch N-NH4+ was significantly affected by NEZ addition in
Nov-2012 (p < 0.05) (138 mg kg−1 vs 50 mg kg−1 of 7CZ and CNTR, respectively) while starting from
May-2013 these differences completely disappeared (p > 0.05). In Jun-2013, there were no differences
between the treatments (p > 0.05) while a relative enrichment of the bulk soil Exch N-NH4+ with
respect to the rhizosphere was apparent in Sep-2013. Generally, at the beginning of the experiment, all treatments showed a low Nmic content, following
the order 7CZ > CNTR > 5NZ = 15NZ, suggesting that zeolitites differentially affected the Nmic
pool. Nmic was increased by NEZ addition while it was decreased by NZ addition. The Nmic pool
increases in May, while it decreases in June after the chemical fertilization; as expected, Nmic content in
rhizospheric soil is higher than that determined in the bulk soil at the harvest (Table S1, Supplementary
Materials). A remarkable increase of Cmic and Nmic and a contemporaneous decrease of N-NO3−in all
the treatments was observed in May-2013 (Figure 4). During the monitored period, Cmic ranged from 16 to 432 mg kg−1. At the beginning of the
experiment large differences (p < 0.05) were detected among the various treatments. In this period, Cmic
content decreases following the order CNTR (148 mg kg−1) >15NZ (98.5 mg kg−1) > 7CZ (43.2 mg kg−1)
> 5NZ (16.1 mg kg−1). 3. Results
able 1). Mo Average N and C pools of NH4+-enriched zeolitite (NEZ). Standard deviation within brackets. brackets. le 1. Average N and C pools of NH4+-enriched zeolitite (NEZ). Standard deviation within brackets. TN
g·kg−1
5.84 (0.90)
Fix N-NH4+
mg·kg−1
343 (21)
Exch N-NH4+
mg·kg−1
4884 (330)
N-NO3−
mg·kg−1
148 (7.4)
Nmic
mg·kg−1
876 (152)
TOC
g·kg−1
10.3 (0.1)
Cmic
mg·kg−1
117 (1)
C/N
1.78
Cmic/Nmic
0.13
brackets. TN
g·kg−1
5.84 (0.90)
Fix N-NH4+
mg·kg−1
343 (21)
Exch N-NH4+
mg·kg−1
4884 (330)
N-NO3−
mg·kg−1
148 (7.4)
Nmic
mg·kg−1
876 (152)
TOC
g·kg−1
10.3 (0.1)
Cmic
mg·kg−1
117 (1)
C/N
1.78
Cmic/Nmic
0.13
Figure 3. Mean soil pH for each treatment. Error bars represent standard deviation. Figure 3. Mean soil pH for each treatment. Error bars represent standard deviation. Fi
3 M
il
H f
h
E
b
d
d d
i
i Figure 3 Mean soil pH for each treatment Error bars represent standard devia
Figure 3. Mean soil pH for each treatment. Error bars represent standard deviation. Appl. Sci. 2019, 9, 4524 8 of 19 Table 2. Average of exchangeable bases and cation exchange capacity (CEC) of soil samples, both
bulk soil and rhizosphere. Standard deviation within brackets. Significant (p < 0.05) or non-significant
(p > 0.05) differences between treatments at each sampling time are indicated by capital letters. Ca2+
K+
Mg2+
Na+
CEC
Time
Treatment
cmol·kg−1
cmol·kg−1
cmol·kg−1
cmol·kg−1
cmol·kg−1
Sep-2013 Rhizo
CNTR
18.3 (0.7) A
8.3 (1.7) A
14.4 (1.2) A
1.8 (0.7) AB
42.8 (4.0) A
5NZ
18.6 (0.6) A
9.9 (0.8) A
17.2 (0.9) B
1.3 (0.1) A
47.1 (1.9) A
15NZ
17.7 (0.9) A
10.4 (0.8) A
17.9 (1.4) B
1.6 (0.6) A
47.5 (2.5) A
7CZ
19.4 (1.1) A
7.7 (3.8) A
15.9 (0.6) AB
3.0 (0.4) B
46.0 (2.7) A
Sep-2013 Bulk
CNTR
19.9 (1.3) A
6.1 (3.3) A
16.8 (1.3) A
2.5 (1.1) AB
45.3 (2.3) A
5NZ
20.3 (0.01) AB
4.9 (0.1) A
21.9 (0.1) B
2.0 (0.02) A
49.1 (0.04) AB
15NZ
23.0 (0.02) B
5.9 (0.004) A
19.5 (0.1) AB
1.7 (0.1) A
50.2 (0.2) B
7CZ
22.0 (0.2) AB
5.0 (0.1) A
19.9 (0.2) B
3.6 (0.04) B
50.6 (0.6) B
3 1 Nitrogen and Carbon Pools in Soil 3.1. Nitrogen and Carbon Pools in Soil 3.1. Nitrogen and Carbon Pools in Soil In May‐2013 a general depletion of N‐NO3− pool is noticed in all
the treatments, increasing in Jun-2013 after urea addition especially in CNTR parcel. N-NO3− values
of ZT (especially 5NZ and 15NZ) were remarkably lower in this period with respect to the CNTR (p
< 0.05) (Table S1, Supplementary materials). Exch N-NH4+ was significantly affected by NEZ addition
in Nov-2012 (p < 0.05) (138 mg kg−1 vs 50 mg kg−1 of 7CZ and CNTR, respectively) while starting from
May-2013 these differences completely disappeared (p > 0.05). In Jun-2013, there were no differences
between the treatments (p > 0.05) while a relative enrichment of the bulk soil Exch N-NH4+ with
respect to the rhizosphere was apparent in Sep-2013. Generally, at the beginning of the experiment, all treatments showed a low Nmic content,
following the order 7CZ > CNTR > 5NZ = 15NZ, suggesting that zeolitites differentially affected the
Nmic pool. Nmic was increased by NEZ addition while it was decreased by NZ addition. The Nmic pool
increases in May, while it decreases in June after the chemical fertilization; as expected, Nmic content
in rhizospheric soil is higher than that determined in the bulk soil at the harvest (Table S1,
Supplementary materials). A remarkable increase of Cmic and Nmic and a contemporaneous decrease
of N-NO3− in all the treatments was observed in May-2013 (Figure 4). Appl. Sci. 2019, 9, x FOR PEER REVIEW
9 of 19
During the monitored period, Cmic ranged from 16 to 432 mg kg-1. At the beginning of the
experiment large differences (p < 0.05) were detected among the various treatments. In this period,
Cmic content decreases following the order CNTR (148 mg kg−1) >15NZ (98.5 mg kg−1) > 7CZ (43.2 mg
kg−1) > 5NZ (16 1 mg kg−1) In May 2013 C
i increased in all treatments except for 15NZ and Figure 4. Nmic (columns) and N-NO3- (lines) content of soil samples across the monitoring period. Figure 4. Nmic (columns) and N-NO3−(lines) content of soil samples across the monitoring period. decreased again in Jun-2013. As expected, Cmic content in the rhizosphere is higher than that
determined in the bulk soil. The average Cmic/Nmic ratio ranges from 3.3 to 94 (Figure 5). 3.1. Nitrogen and Carbon Pools in Soil In May-2013, Cmic increased in all treatments, except for 15NZ, and decreased
again in Jun-2013. As expected, Cmic content in the rhizosphere is higher than that determined in the
bulk soil. The average Cmic/Nmic ratio ranges from 3.3 to 94 (Figure 5). In Nov-2012, low values of
Cmic/Nmic were observed in 5NZ and 7CZ treatments; concerning 5NZ, the low ratio is due to the
lower Cmic with respect to the other treatments, while for 7CZ the low ratio may have been due to the
high Nmic. All ZT have a higher Cmic/Nmic ratio than CNTR after May-2013 because of the lower Nmic;
on average, Cmic/Nmic ratio increased in order of 15NZ > 5NZ > 7CZ > CNTR. TOC showed significant
differences between the treatments and among the sampling times (p < 0.05). In particular, a depletion
of TOC content was observed in 7CZ in Nov-2012 and in Jun-2013, after urea addition (Table S1, 9 of 19
t is
O3- Appl. Sci. 2019, 9, 4524
values (p < 0.0
significantly lo Supplementary Materials). As expected, TOC content in Sep-2013 was higher in rhizosphere than in
the bulk soil and there was a significant correlation between TN and TOC (ρ = 0.83, p < 0.05). Soil C/N
ratio showed significant differences among the treatments only at the beginning of the experiment
because of the higher TN of 7CZ (p < 0.05), while during the rest of the monitored period, soil C/N
ratio remained constant around 10 (Table S1, Supplementary Materials). HS C content was reported in
Table 3. TEC content ranged from 8.3 to 19.2 g kg−1 (data not shown), whereas the extraction ratio
ranged between 41 to 87%. Notably, the highest extraction ratio was observed in 15NZ parcel, followed
by 5NZ. Generally, HA C content is higher than that determined in FA, with exception of 7CZ in
Nov-2012, in which the FA/HA ratio is higher than 1. A low HI was generally recorded in all the
treatments with the exception of 5NZ, where especially in May, June and Spet-2013 Bulk, HI increases
through values above 0.5, highlighting a prevalence of NHS. The HD% generally varied between 30%
and 50%, with the exception of 15NZ that showed large variations (from 27.8% in Nov-2012 to 68.2% in
May-2013). and 15NZ, respectively) (p < 0.05). 3.1. Nitrogen and Carbon Pools in Soil In Nov-2012,
low values of Cmic/Nmic were observed in 5NZ and 7CZ treatments; concerning 5NZ, the low ratio is
due to the lower Cmic with respect to the other treatments, while for 7CZ the low ratio may have been
due to the high Nmic. All ZT have a higher Cmic/Nmic ratio than CNTR after May-2013 because of the
lower Nmic; on average, Cmic/Nmic ratio increased in order of 15NZ > 5NZ > 7CZ > CNTR. TOC showed
significant differences between the treatments and among the sampling times (p < 0.05). In particular,
a depletion of TOC content was observed in 7CZ in Nov-2012 and in Jun-2013, after urea addition
(Table S1, Supplementary materials). As expected, TOC content in Sep-2013 was higher in
rhizosphere than in the bulk soil and there was a significant correlation between TN and TOC (ρ=0.83,
p < 0.05). Soil C/N ratio showed significant differences among the treatments only at the beginning
of the experiment because of the higher TN of 7CZ (p < 0.05), while during the rest of the monitored
period, soil C/N ratio remained constant around 10 (Table S1, Supplementary materials). HS C
content was reported in Table 3. TEC content ranged from 8.3 to 19.2 g kg−1 (data not shown), whereas
the extraction ratio ranged between 41 to 87%. Notably, the highest extraction ratio was observed in
15NZ parcel, followed by 5NZ. Generally, HA C content is higher than that determined in FA, with
exception of 7CZ in Nov-2012, in which the FA/HA ratio is higher than 1. A low HI was generally
recorded in all the treatments with the exception of 5NZ, where especially in May, June and Spet-
2013 Bulk, HI increases through values above 0.5, highlighting a prevalence of NHS. The HD%
generally varied between 30% and 50%, with the exception of 15NZ that showed large variations
(from 27.8% in Nov-2012 to 68.2% in May-2013). Figure 4. Nmic (columns) and N-NO3- (lines) content of soil samples across the monitoring period. Figure 4. Nmic (columns) and N-NO3−(lines) content of soil samples across the monitoring period. generally varied between 30% and 50%, with the exception of 15NZ that showed large variations
(from 27.8% in Nov-2012 to 68.2% in May-2013). Figure 5. Microbial biomass C/N ratio over the monitoring period. Figure 5. Microbial biomass C/N ratio over the monitoring period. Figure 5. Microbial biomass C/N ratio over the monitoring period. Figure 5. 3.2. Total N and C Content in Sorghum Plant Components No significant differences in TN and TOC content were detected in the sorghum organs
(e.g., roots, stems, leaves and panicles) among the treatments (p > 0.05) (Table S2, Supplementary
Materials). Generally, leaves and panicles had a higher TN content than roots and stems, while
TOC was similar in all the plant organs, resulting in decline of the C/N ratio following the order
stems > roots > panicles > leaves. 3.1. Nitrogen and Carbon Pools in Soil Microbial biomass C/N ratio over the monitoring period. Appl. Sci. 2019, 9, 4524 10 of 19 Table 3. C content of Humic Acids (HA) and Fulvic Acids (FA), percentage of Total Extractable and Total
Organic Carbon (TEC/TOC) ratio, Humification Index (HI) and Humification Degree (HD) according to
Ciavatta et al. [33] and Sequi et al. [34]. HA
FA
TEC/TOC
HI
HD
Time
Treatment
g kg−1
g kg−1
%
%
Nov-2012
CNTR
5.3
3.4
47.4
0.4
34.4
5NZ
4.3
5.0
46.2
0.2
37.7
15NZ
3.9
3.7
41.4
0.5
27.8
7CZ
2.7
3.9
40.1
0.3
31.9
May-2013
CNTR
7.4
3.2
63.0
0.5
43.1
5NZ
8.6
3.5
74.4
0.6
46.9
15NZ
11.4
3.6
87.3
0.3
68.2
7CZ
5.9
2.5
48.6
0.1
43.8
Jun-2013
CNTR
4.5
3.1
52.3
0.4
38.6
5NZ
4.3
2.9
65.0
0.8
35.5
15NZ
3.8
3.1
50.6
0.3
39.2
7CZ
3.4
3.0
43.6
0.6
27.4
Sep-2013 Bulk
CNTR
5.9
3.2
52.9
0.2
44.2
5NZ
3.9
2.9
54.5
0.9
29.4
15NZ
8.6
3.3
70.7
0.4
51.3
7CZ
5.9
3.5
57.9
0.4
41.2
Sep-2013 Rhizo
CNTR
8.1
3.1
58.1
0.3
43.4
5NZ
6.9
3.2
48.5
0.4
34.7
15NZ
9.7
3.2
49.2
0.2
41.0
7CZ
7.1
3.3
48.8
0.3
37.0 3.2. Total N and C Content in Sorghum Plant Components 3.3. δ15N Natural Abundance in Soil and Sorghum Organs TN and Fix N-NH4+ δ15N differs significantly (p < 0.05) among treatments and time (Table 4). At the beginning of the experiment (Nov-2012), 7CZ shows a remarkably higher isotopic signature
(δ15N of 7.46% and 8.27% for TN and Fix N-NH4+, respectively) with respect to the other treatments
(p < 0.05). Furthermore, without considering 7CZ treatment, Fix N-NH4+ pool showed a high δ15N
variability (from 4.55% to 6.00%) with respect to the TN pool (from 3.38% to 3.86%). In May-2013,
TN δ15N is still significantly higher (p < 0.05) in 7CZ treatment, while no significant differences were
observed regarding Fix N-NH4+ pool. In Jun-2013 (after the chemical fertilization), all treatments
showed a homogeneous δ15N in both TN and Fix N-NH4+ pools (p > 0.05). In Sep-2013, rhizosphere
samples show higher δ15N with respect to the bulk soils in both TN and Fix N-NH4+ pools (especially
in the latter). The isotopic signatures of NEZ, urea and di-ammonium phosphate were: +18.4 (± 0.5) %,
−1.18 (± 0.06) % and −0.89 (± 0.09) %, respectively (Table 5). Isotopic analysis on the harvested plants
indicated that the δ15N of plant tissues was influenced by the different agricultural managements
employed in the experimentation (Table 5). δ15N was significantly higher in 7CZ plants leaves and
significantly lower in all 5NZ plants organs with respect to the CNTR plants (p < 0.05). In Figure 6,
the TN vs δ15N of the different plants’ organs are plotted together with urea, NEZ and bulk soil. It is
evident that the 7CZ leaves are strongly affected by NEZ contribution, while 5NZ plants are more
influenced by a low δ15N N-source. Appl. Sci. 2019, 9, 4524 11 of 19 11 of 19 3.4. Sorghum Yield The CNTR parcel returned a yield of 5818 kg ha−1. The two NZ returned 6032 kg ha−1 each, while
7CZ parcel gave 6627 kg ha−1. Compared with the CNTR, the three zeolitite-treated parcels gained
from 3.7 to 13.9%, although the yield results are not statistically significant. The CNTR parcel returned a yield of 5818 kg ha−1. The two NZ returned 6032 kg ha−1 each, while
7CZ parcel gave 6627 kg ha−1. Compared with the CNTR, the three zeolitite-treated parcels gained
from 3.7 to 13.9%, although the yield results are not statistically significant. Table 4. Average 15N natural abundance in soil N pools (TN and Fix N-NH4+). Standard deviation
within brackets. Significant (p < 0.05) or non-significant (p > 0.05) differences between treatments at
each sampling time are indicated by capital letters. “*” and “x” symbols indicates the presence or the
absence, respectively, of a significant difference among the variables for Treatments and Time in the
experiment period and correlating treatments with time (Treatment*Time). TN δ15N vs air
Fix δ15N vs air
Time
Treatment
%
%
Nov-2012
CNTR
3.86 (0.15) A
6.00 (0.17) B
5NZ
3.83 (0.22) A
5.28 (0.45) AB
15NZ
3.38 (0.19) A
4.55 (0.31) A
7CZ
7.46 (0.53) B
8.27 (0.36) C
May-2013
CNTR
3.75 (0.20) AB
3.34 (0.15) A
5NZ
3.34 (0.06) A
3.38 (0.30) A
15NZ
3.31 (0.62) A
3.84 (0.04) A
7CZ
4.87 (0.17) B
3.11 (0.12) A
Jun-2013
CNTR
3.11 (0.28) A
3.65 (0.43) A
5NZ
3.37 (0.10) A
3.38 (0.17) A
15NZ
3.32 (0.05) A
3.46 (0.13) A
7CZ
3.27 (0.17) A
3.73 (0.19) A
Sep-2013 Bulk
CNTR
2.68 (0.11) A
1.37 (0.11) A
5NZ
2.39 (0.09) A
1.83 (0.23) AB
15NZ
2.73 (0.14) A
1.79 (0.03) AB
7CZ
2.55 (0.29) A
2.09 (0.16) B
Sep-2013 Rhizo
CNTR
4.22 (0.38) A
2.47 (0.26) A
5NZ
4.17 (0.31) A
2.81 (0.75) A
15NZ
4.55 (1.11) A
2.40 (0.46) A
7CZ
4.45 (0.19) A
2.52 (0.31) A
NEZ
18.4 (0.5)
17.3 (0.4)
Urea
−1.18 (0.06)
-
Diammonium
Phosphate
−0.89 (0.09)
-
Treatment
*
*
Time
*
*
Treatment*Time
*
*
le 5
Average 15N natural abundance in sorghum organs at the harvest
SD within brac Table 5. Average 15N natural abundance in sorghum organs at the harvest. SD within brackets. Significant (p < 0.05) or non-significant (p > 0.05) differences between treatments for each organ are
indicated by capital letters. Table 5. 3.4. Sorghum Yield Average 15N natural abundance in sorghum organs at the harvest. SD within brackets. Significant (p < 0.05) or non-significant (p > 0.05) differences between treatments for each organ are
indicated by capital letters. δ15N vs Air
Plant Organ
Treatment
%
Roots
CNTR
10.2 (1.4) AB
5NZ
6.33 (1.69) A
15NZ
10.3 (2.8) AB
7CZ
11.6 (2.7) B 12 of 19 Appl. Sci. 2019, 9, 4524 Table 5. Cont. δ15N vs Air
Plant Organ
Treatment
%
Stems
CNTR
9.95 (0.81) B
5NZ
6.08 (1.56) A
15NZ
11.1 (1.1) B
7CZ
11.0 (1.5) B
Leaves
CNTR
12.3 (1.6) B
5NZ
8.88 (0.94) A
15NZ
13.8 (2.2) BC
7CZ
18.1 (1.7) C
Panicles
CNTR
10.3 (0.4) B
5NZ
6.82 (0.97) A
15NZ
10.0 (1.8) B
7CZ
11.7 (2.0) B 4.1.1. NEZ Effects on N Pool Size and Isotopic Signature 4.1.1. NEZ Effects on N Pool Size and Isotopic Signature 4.1.1. NEZ Effects on N Pool Size and Isotopic Signature The addition of NEZ into the soil (7CZ treatment) did not lead to significant differences in TN or Fix
N-NH4+ content regardless the N input of about 410 kg of N ha−1 brought by NEZ introduction. On the
other hand, this N input has clearly influenced Exch N-NH4+, N-NO3−and Nmic pools, increasing their
size. Through the application of a simple mass balance equation, it was estimated that the increment
in 7CZ Exch N-NH4+ pool corresponded to the amount of N introduced with NEZ in the first 30 cm
of soil. As it is known, NH4+-enriched zeolites can act as slow-released fertilizer [37–39] and in our
experiment this behavior was clarified by the isotopic signature of the TN pool during the agricultural
management. The δ15N increase of TN pool in 7CZ with respect to that found in the other treatments
is in fact due to the N isotopic signature of the pig slurry contained into the NEZ. Dittert et al. [40]
found a δ15N of 5.90% in agricultural soil treated with slurry while a δ15N of 3.90% was found in
soils treated with inorganic fertilizers. Pig slurry usually has high δ15N values, as a result of the
fractionation processes occurring during volatilization of NH3 after the excretion, which causes an
enrichment in the heavier 15N atoms in the substrate (slurry) with respect to the product (ammonia
gas) [40–44]. The significantly higher δ15N of TN in May-2013 suggested that a significant amount
of N introduced with NEZ was still present in the soil before the sowing. At the beginning of the
experimentation, the organic C cycle of 7CZ treatment was affected by the high N input provided by
NEZ, resulting in a decrease of TOC content and C/N ratio; furthermore, an increase of FA fraction can
be interpreted as an increase in depolymerization processes. 4. Discussion As expected, CEC was generally higher in ZT with respect to the CNTR, especially in the bulk soil. Ozbahce et al. [36] and Gholamhoseini et al. [20] found a similar tendency after mixing zeolitite in the
first 30 cm of soil or after the spreading in combination with cattle manure. Both bulk and rhizosphere
soils in the 7CZ parcel showed higher exchangeable Na+ with respect to the other treatments, likely
because of the NEZ enrichment process with pig slurry. 4.1. NEZ Influence on Soil N and C Pools. 4.1. NEZ Influence on Soil N and C Pools. 4.1.3. NEZ Specific Effects on Fixed N-NH4+ Pool A possible hypothesis is that Fix N-NH4+ pool is in dynamic equilibrium with exchangeable and
soluble N-NH4+ pools [52,53]. In our case, this hypothesis is supported by the δ15N turnover of fixed
pool that suggested a continuous N replacement during the monitoring period. At the start of the
experimentation, Fix δ15N of CNTR and NZ treatments was probably still affected by previous slurry
treatments as shown by the higher isotopic signature. Generally, microbial activity increases with the
increase of soil temperature, affecting the other N pools, in particular, a decrease in N-NO3−and Exch
N-NH4+ pools (as in May-2013 sampling) can alter the equilibrium with fixed interlayer ammonium. This decrease can cause a release of N from Fix N-NH4+ pool in order to maintain equilibrium, with a
continuous N turnover and a consequent change in δ15N. No δ15N turnover of fixed ammonium was
observed immediately after the chemical fertilization applied in Jun-2013, while the addition of urea
was marked by a pH increase [54]. On the contrary, in Sep-2013, a probable storage of N from chemical
fertilization was visible in Fix N-NH4+ pool (and consequently in TN pool), as demonstrated by the
significantly lower δ15N (very close to the δ15N of urea and di-ammonium phosphate). This behavior
is recorded only in the bulk soil, where the exploitation by the plant roots is less important than in
the rhizosphere. 4.1.2. NEZ Specific Effects on Soil Microbial Biomass On the other hand, Nmic increment in 7CZ parcel was not counterbalanced by a parallel Cmic
increment, resulting in a strong decrease of the Cmic/Nmic and likely in an increase of organic matter
(OM) mineralization processes [44]. No evidence of enhanced nitrification was observed in 7CZ parcel Appl. Sci. 2019, 9, 4524 13 of 19 13 of 19 after the zeolitite spreading as N-NO3−content was very similar to the CNTR notwithstanding the
large N input provided by NEZ spreading (with residual N-NO3−from the pig slurry treatment). The agronomic year before the experimental cultivation was low in precipitation and the drought
caused low yield and accumulation of high amounts of N from fertilizers in the upper soil horizon,
thus no differences were expected between CNTR and ZT. A possible explanation of this behavior
is that controlled retention and release capacity of NH4+ contained in the microsite of the open-ring
structure of the mineral. These ions are physically protected and likely less available to nitrifying
bacteria (leading consequently to a lesser N-NO3−content in the pore-waters) [45–48]. This hypothesis
is plausible but not supported by a direct measurement of gross nitrification rates (e.g., by the mean
of 15N pool dilution technique). In this light, further studies are urgently needed where gross N
transformation rates are measured in soil amended with NEZ and NZ to efficiently quantify gross N
production and consumption through mineralization, nitrification and denitrification processes. p
p
g
p
In May-2013, Cmic and Nmic increased significantly in all treatments, while N-NO3−pool
significantly decreased (but not Exch N-NH4+ pool with the exception of 7CZ) (Figure 4), suggesting
that microbial immobilization prevailed over mineralization [49]. In addition to NO3−decrease,
7CZ treatment showed a reduction of Exch N-NH4+ pool, where the N surplus caused by the NEZ
introduction completely disappeared. A fraction of this mineral N depletion can be attributable
to NO3−leaching and/or to N2-N2O losses through denitrification process (probably enhanced by
waterlogged conditions; [25]). It is important to consider that NEZ introduction affected mainly
the Exch N-NH4+ and not the Fix N-NH4+ pool and that 7CZ still had high TN δ15N in May-2013,
confirming that an important fraction of NEZ-N was still present in the soil system. It is thus possible
that a large fraction of the exchangeable N contained into NEZ has been immobilized by the microbial
biomass [50,51]. 4.2. NZ Influence on Soil N and C Pools The introduction of natural zeolitites (5NZ and 15NZ, respectively) affected the soil C and N
pools. The decrease in the total reserve of Fix N-NH4+ in 15NZ in Nov-2012 was probably due to a
dilution effect operated by the high amount of NZ (devoid of N) introduced. Instead, the decrease of
Fix N-NH4+ during the growing season suggest an exploitation by plants, confirming the active role of
this pool in crop nutrition [55,56]. Nevertheless, no significant differences in the amount of Fix N-NH4+
pool were found between the various treatments, suggesting that the N subjected to CEC processes by
zeolitites belongs to the exchangeable pool. The generally lower N-NO3−content in the NZ treatments
may be attributed both to the aforementioned controlled retention of NH4+ by the zeolitite, which can Appl. Sci. 2019, 9, 4524 14 of 19 14 of 19 reduce the availability of N to nitrifying bacteria and to the lower amount of N fertilizer (from 30 to
50%) with respect to CNTR (where N-NO3−pool increased significantly). The introduction of NZ also
affected microbial biomass N and C immobilization. In Nov-2012, lower Cmic was observed in all ZT,
especially in 5NZ, suggesting that NZ introduction cause an initial disequilibrium in microbial biomass. During the sorghum-growing season (Jun-2013), microbial biomass was probably influenced by the
competition with the plants, resulting in a consistent decrease in Cmic and Nmic in all the treatments. Apparently, 15NZ treatment had a stronger influence on microbial biomass N, resulting in a lower N
immobilization that was already visible in May-2013 (in this case even Cmic pool was affected) and
lasted until Sep-2013, suggesting more zeolitite was in the soil the less N was easily available to soil
microorganisms. The C/N ratio of microbial biomass may give information regarding bacterial or
fungal prevalence in the microbial community; usually bacteria have on average a lower C/N than fungi
because of a lower N requirement by the latter [57,58]. Agricultural management (tillage or no-tillage),
nutrient availability, soil pH, temperature and moisture may influence fungal/bacterial ratio [15,59,60]. In this respect, Cmic/Nmic ratio suggested that the introduction of natural zeolitites caused a shift in
microbial population through a greater fungal prevalence [15,59,60] especially in 15NZ. 4.3. Zeolitite Influence on Rhizosphere At the harvest, rhizosphere was depleted in Exch N-NH4+ with respect to bulk soils but
significantly enriched in microbial biomass C-N, TOC, HA and TN. It is reasonable that the decrease
in the exchangeable pool was due to an exploitation by plant roots while bulk soil results relatively
enriched. Rhizosphere is known to be a zone with higher metabolic activity, thus the release of organic
compounds from roots may explain the increase in microbial biomass, TOC, HA and TN [61,62] and
also the slight fractionation observed in TN and Fix N-NH4+ pools δ15N. It is interesting to note that
at the end of the growing season, rhizosphere TOC was probably influenced by zeolitite addition
(especially in 15NZ), showing an increase with respect to the CNTR. This evidence suggests a higher
efficiency of microbial biomass, but this is not supported by a parallel increment in Cmic and Nmic
with respect to CNTR. On the other hand, Ozbahce et al. [36] found that the application of NZ mixed
in the first 30cm soil layer increased OM content, although a clear explanation of this phenomenon
was not given. Italian zeolitic tuffs can adsorb a significant amount of HS from solutions, especially
when divalent cations (such as Ca2+) act as bridge between the negatively charged mineral surface and
the organic phase [63]. A plausible hypothesis is that zeolitite can adsorb HS reducing in this way
organic C losses and increasing thus TOC in the soil [64]. Ca2+ is indeed the major exchangeable base
of the studied NZ and it can act as bridge for the sorption of HS in zeolitic tuff. Further studies are
required to confirm this very important aspect. 4.2. NZ Influence on Soil N and C Pools A greater
fungal/bacteria biomass ratio can be often interpreted as marker of a more sustainable agricultural
system, where the nutrients required for the plant growth are supplied by OM decomposition and
N mineralization with probably lesser N loss in the environment [15,59,60]. It is thus likely that the
presence of zeolites caused a lower N accessibility into the N soil pools, making it less available to soil
microorganisms and favoring the development of fungal biomass. These implications on microbial
biomass affect the soil C cycle, where a lower HI and a greater FA fraction content can contribute to
lower C immobilization. 4.4. Plant-Zeolitite Interactions N is preferentially allocated in leaves and panicles and the lack of significant differences in
N content with respect to the distinct treatments suggests that, notwithstanding the fertilization
reductions, plants uptake similar N amounts, reflecting a higher fertilizer recovery or N use efficiency. As reported in literature [65,66] δ15N of leaves is determined by the physiological mechanism within
the plants and by the isotopic ratio of external N sources. Moreover, intra-plant N isotopic variations,
between roots and leaves can be due to different patterns of N assimilation or reallocation. These
intra-plant δ15N variations were recognizable in the sorghum plants, where leaves δ15N was generally Appl. Sci. 2019, 9, 4524 15 of 19 greater (from 2 to 3%) than that determined in roots, except for 7CZ where differences were even
greater (about 6%). Many authors used 15N natural abundance of leaves in order to trace the N
source in the soil [65–69] mostly because leaves represent the primary sink of N and the largest plant
N pool [41]. Average δ15N of 7CZ leaves was very close to NEZ δ15N and significantly higher than
the leaves from the other parcels (Figure 6). The higher isotopic signature of 7CZ leaves indicated
that during the growing season, an uptake from a significantly higher δ15N occurred. Assuming
negligible fractionation effects during uptake, the main N source was probably represented by NEZ-N. Indeed, 7CZ is the only parcel where a high δ15N input (represented by pig slurry) was added during
the experimentation. It has been demonstrated that crops grown under chemical fertilizers (such as
urea) have lower δ15N with respect to plants grown under organic fertilizers (which have usually
higher δ15N) [70–72]. It is important to consider that the other N inputs were urea and di-ammonium
phosphate, which have a negative δ15N and were applied to all treatments, supporting the hypothesis
that NEZ was an efficient slow release fertilizer and retained N even after several months and with
no important N losses occurring (from leaching or denitrification). Most N in soil is bound in forms
not immediately available to plants [41,71], so usually the δ15N of TN is not a good approximation of
the isotopic signature of the N source preferentially used by plants. Anyhow, 15N natural abundance
in leaves of CNTR, 15NZ and especially 5NZ reflect the use of N characterized by a lower δ15N and
related to chemical fertilizers signature. 4.4. Plant-Zeolitite Interactions It is important to note that notwithstanding the lower urea
application (−30% with respect to the CNTR), in 5NZ and 15NZ treatments, leaves δ15N is similar
or even lower with respect to CNTR. In particular, as visible from the mixing model (Figure 6), 5NZ
plants showed the lowest δ15N not only in leaves but also in roots, stems and panicles, suggesting a
higher efficiency in the uptake of N from a source with lower δ15N, i.e., chemical fertilizers. Appl. Sci. 2019, 9, x FOR PEER REVIEW
15 of 19
denitrification). Most N in soil is bound in forms not immediately available to plants [41,71], so
usually the δ15N of TN is not a good approximation of the isotopic signature of the N source
preferentially used by plants. Anyhow, 15N natural abundance in leaves of CNTR, 15NZ and
especially 5NZ reflect the use of N characterized by a lower δ15N and related to chemical fertilizers
signature. It is important to note that notwithstanding the lower urea application (−30% with respect
to the CNTR), in 5NZ and 15NZ treatments, leaves δ15N is similar or even lower with respect to
CNTR. In particular, as visible from the mixing model (Figure 6), 5NZ plants showed the lowest δ15N
not only in leaves but also in roots, stems and panicles, suggesting a higher efficiency in the uptake
f N f
ith l
δ15N i
h
i
l f
tili The positive yield results of all ZT must be attributed also to the soil physical properties
amelioration induced by zeolitite amendments. In our case, zeolitites can in fact increase soil water
retention and, thanks to their coarser size with respect to the fine-grained soil (particle-size analysis
performed within ZeoLIFE project are not shown in this work) they can also increase soil permeability
and aeration [28,36]. of N from a source with lower δ15N, i.e. chemical fertilizers. The positive yield results of all ZT must be attributed also to the soil physical properties
amelioration induced by zeolitite amendments. In our case, zeolitites can in fact increase soil water
retention and, thanks to their coarser size with respect to the fine-grained soil (particle-size analysis
performed within ZeoLIFE project are not shown in this work) they can also increase soil
permeability and aeration [28,36]. Figure 6. δ15N vs total nitrogen (TN) mixing diagram for sorghum plant organs. Chemical fertilizers,
NEZ and Soil have been chosen as end-members. Figure 6. 4.4. Plant-Zeolitite Interactions δ15N vs total nitrogen (TN) mixing diagram for sorghum plant organs. Chemical fertilizers,
NEZ and Soil have been chosen as end-members. Figure 6. δ15N vs total nitrogen (TN) mixing diagram for sorghum plant organs. Chemical fertilizers,
NEZ and Soil have been chosen as end-members. Figure 6. δ15N vs total nitrogen (TN) mixing diagram for sorghum plant organs. Chemical fertilizers,
NEZ and Soil have been chosen as end-members. Appl. Sci. 2019, 9, 4524 16 of 19 16 of 19 5. Conclusions Funding: This work has been sustained by the EU-funded ZeoLIFE project (LIFE+10/ENV/IT000321). Funding: This work has been sustained by the EU-funded ZeoLIFE project (LIFE+10/ENV/IT000321). Acknowledgments: We gratefully acknowledge Francesco Droghetti for his help during sampling procedures,
Paola Gioacchini for her support in EA-IRMS analyses and Marco Natale for his help in laboratory runs and
anonymous reviewers for the valuable comments and suggestions. Conflicts of Interest: The authors declare no conflict of interest. References . Conway, G. The Double Green Revolution: Food for All in the 21st Century; Penguin: London, UK, 1997; p. 1. Conway, G. The Double Green Revolution: Food for All in the 21st Century; Penguin: London, UK, 1997; p. 334. 2. Smil, V. Nitrogen in crop production: An account of global flows. Glob. Biogeochem. Cycles 1999, 13, 647–662. [CrossRef] 2. Smil, V. Nitrogen in crop production: An account of global flows. Glob. Biogeochem. Cycles 1999, 13, 647–662. [CrossRef] 2. Smil, V. Nitrogen in crop production: An account of global flows. Glob. Biogeochem. Cycles 1999, 13, 647–662. [CrossRef] 3. Tilman, D.; Fargione, J.; Wolff, B.; D’Antonio, C.; Dobson, A.; Howarth, R.; Schindler, D.; Schlesinger, W.H.;
Simberloff, D.; Swackhamer, D. Forecasting agriculturally driven global environmental change. Science 2001,
292, 281–284. [CrossRef] [PubMed] 3. Tilman, D.; Fargione, J.; Wolff, B.; D’Antonio, C.; Dobson, A.; Howarth, R.; Schindler, D.; Schlesinger, W.H.;
Simberloff, D.; Swackhamer, D. Forecasting agriculturally driven global environmental change. Science 2001,
292, 281–284. [CrossRef] [PubMed] 4. Oenema, O.; Kros, H.; Vries, W. Approaches and uncertainties in nutrient budgets: Implications for nutrient
management and environmental policies. Eur. J. Agron. 2003, 20, 3–16. [CrossRef] 4. Oenema, O.; Kros, H.; Vries, W. Approaches and uncertainties in nutrient budgets: Implications for nutrient
management and environmental policies. Eur. J. Agron. 2003, 20, 3–16. [CrossRef] 5. Kroeze, C.; Aerts, R.; Breemen, N.; Dam, D.; Hoek, K.; Hofschreuder, P.; Hoosbeek, M.; Klein, J.; Kros, H.;
Oene, H.; et al. Uncertainties in the fate of nitrogen I: An overview of sources of uncertainty illustrated with
a Dutch case study. Nutr. Cycl. Agroecosyst. 2003, 66, 43–69. [CrossRef] 6. Passaglia, E. Zeoliti Naturali, Zeolititi e Loro Applicazioni; Arvan: Padova, Italy, 2008; p. 104. 7. Wu, Q.; Chi, D.; Xia, G.; Chen, T.; Sun, Y.; Song, Y. Effects of Zeolite on Drought Resistance and Water—Nitrogen
Use Efficiency in Paddy Rice. J. Irrig. Drain. Eng. 2019, 145, 4019024. [CrossRef] 7. Wu, Q.; Chi, D.; Xia, G.; Chen, T.; Sun, Y.; Song, Y. Effects of Zeolite on Drought Resistance and Water—Nitrogen
Use Efficiency in Paddy Rice. J. Irrig. Drain. Eng. 2019, 145, 4019024. [CrossRef] 8. Wu, Q.; Chen, T.; Chi, D.; Xia, G.; Sun, Y.; Song, Y. Increasing nitrogen use efficiency with lower nitrogen
application frequencies using zeolite in rice paddy fields. Int. Agrophys. 2019, 33, 263–269. [CrossRef] 8. Wu, Q.; Chen, T.; Chi, D.; Xia, G.; Sun, Y.; Song, Y. 5. Conclusions In this work, a detailed investigation of the N-C pools and δ15N dynamics in the soil-sorghum
system were studied in a field amended with natural and NH4+-enriched zeolitites, under fertilization
reductions. ZT generally had a weak influence on the total reserve of Fix N-NH4+ pool. However, its
reserve decreased during the growing season and a δ15N turnover occurs, confirming both its active
role in plant nutrition and its dynamic equilibrium with the other soil N pools. Moreover, Fix δ15N
turnover indicate that part of N from chemical fertilizers was probably stored in this pool at the end
of the agronomic year. The N introduced in the soil system with NEZ increased the Exch N-NH4+
reserve, suggesting that N adsorbed by zeolitites preferentially affected this pool. ZT (especially NZ)
influenced the N-NO3−pool, resulting in lower nitrate contents suggesting a possible nitrification and
denitrification rate reduction. Microbial activity was strongly affected by NEZ and NZ introduction
which seems to cause at first a disorder, then a possible change in microbial population towards a fungal
prevalence, usually indicative of a system with lower N accessible to microorganisms. 15N natural
abundance in soils and plants, together with the positive yield results, suggested that plants benefited
from N NEZ. However, in all likelihood, plants did not directly exploit N from NEZ, but rather the
transfer involves complex interactions among soil microbial biomass. Further investigations focused on
this biotic mediation are needed. Most of the N-C pools dynamics were not affected by NZ amendments
but, in all likelihood, an increase in fertilizers recovery by plants occurred. The spreading of zeolitites
may improve soil physical-chemical properties, enhance nutrient use efficiency as well as augment
economical and an environmental saving. Supplementary Materials: The following are available online at http://www.mdpi.com/2076-3417/9/21/4524/s1,
Table S1: Average of N and C pools. Table S2: Average of TN, TOC and C/N of sorghum organs. Supplementary Materials: The following are available online at http://www.mdpi.com/2076-3417/9/21/4524/s1,
Table S1: Average of N and C pools. Table S2: Average of TN, TOC and C/N of sorghum organs. Author Contributions: G.F., L.V.A. and M.C. conceptualized the research project. G.F., L.V.A., B.F. and D.D.G.,
designed the study and analyzed the data. All authors contributed to the writing of the manuscript. Author Contributions: G.F., L.V.A. and M.C. conceptualized the research project. G.F., L.V.A., B.F. and D.D.G.,
designed the study and analyzed the data. All authors contributed to the writing of the manuscript. .
Li, Z.; Zhang, Y.; Li, Y. Zeolite as slow release fertilizer on spinach yields and quality in a greenhouse
J. Plant Nutr. 2013, 36, 1496–1505. [CrossRef] .
Wu, Q.; Chen, T.; Chi, D.; Xia, G.; Sun, Y.; Song, Y. Increasing nitrogen use efficiency with lower nitro
application frequencies using zeolite in rice paddy fields. Int. Agrophys. 2019, 33, 263–269. [CrossRef] References CN elemental and isotopic
investigation in agricultural soils: Insights on the effects of zeolitite amendments. Chem. Der Erde 2017, 77, 18. Ferretti, G.; Keiblinger, K.M.; Zimmermann, M.; Di Giuseppe, D.; Faccini, B.; Colombani, N.; Mastrocicco, M.;
Zechmeister-Boltenstern, S.; Mentler, A. High resolution short-term investigation of soil CO2, N2O, NOx and
NH3 emissions after different chabazite zeolite amendments. Appl. Soil Ecol. 2017, 119, 138–144. [CrossRef] 19. Ferretti, G.; Di Giuseppe, D.; Natali, C.; Faccini, B.; Bianchini, G.; Coltorti, M. CN elemental and isotopic
investigation in agricultural soils: Insights on the effects of zeolitite amendments. Chem. Der Erde 2017, 77,
45–52. [CrossRef] 20. Gholamhoseini, M.; Ghalavand, A.; Khodaei-Joghan, A.; Dolatabadian, A.; Zakikhani, H.; Farmanbar, E. Zeolite-amended cattle manure effects on sunflower yield, seed quality, water use efficiency and nutrient
leaching. Soil Tillage Res. 2013, 126, 193–202. [CrossRef] g
g
21. Di Giuseppe, D.; Ferretti, G.; Faccini, B.; Blasi, E.; Passeri, N.; Bianchini, G.; Coltorti, M. Is it possible to
cultivate corn in a sustainable way using a quarry waste? Period. Mineral. 2016, 85, 179–183. 22. Arcara, P.G.; Gamba, C.; Bidini, D.; Marchetti, R. The effect of urea and pig slurry fertilization on denitrification,
direct nitrous oxide emission, volatile fatty acids, water-soluble carbon and anthrone-reactive carbon in
maize-cropped soil from the Po plain (Modena, Italy). Biol. Fertil. Soils 1999, 29, 270–276. [CrossRef] 23. Faccini, B.; Di Giuseppe, D.; Malferrari, D.; Coltorti, M.; Abbondanzi, F.; Campisi, T.; Laurora, A.; Passaglia, E. Ammonium-exchanged zeolitite preparation for agricultural uses: From laboratory tests to large-scale
application in ZeoLIFE project prototype. Period. Mineral. 2015, 84, 303–321. 24. Faccini, B.; Di Giuseppe, D.; Ferretti, G.; Coltorti, M.; Colombani, N.; Mastrocicco, M. Natural and
NH4+-enriched zeolitite amendment effects on nitrate leaching from a reclaimed agricultural soil (Ferrara
Province, Italy). Nutr. Cycl. Agroecosyst. 2018, 110, 327–341. [CrossRef] 25. Mastrocicco, M.; Colombani, N.; Di Giuseppe, D.; Faccini, B.; Coltorti, M. Contribution of the subsurface
drainage system in changing the nitrogen speciation of an agricultural soil located in a complex marsh
environment (Ferrara, Italy). Agric. Water Manag. 2013, 119, 144–153. [CrossRef] 26. Di Giuseppe, D.; Faccini, B.; Mastrocicco, M.; Colombani, N.; Coltorti, M. Reclamation influence and
background geochemistry of neutral saline soils in the Po River Delta Plain (Northern Italy). Environ. Earth Sci. 2014, 72, 2457–2473. [CrossRef] 27. IUSS Working Group. World Reference Base for Soil Resources. References Increasing nitrogen use efficiency with lower nitrogen
application frequencies using zeolite in rice paddy fields. Int. Agrophys. 2019, 33, 263–269. [CrossRef] 9. Li, Z.; Zhang, Y.; Li, Y. Zeolite as slow release fertilizer on spinach yields and quality in a greenhouse test. J. Plant Nutr. 2013, 36, 1496–1505. [CrossRef] 9. Li, Z.; Zhang, Y.; Li, Y. Zeolite as slow release fertilizer on spinach yields and quality in a greenhouse test. J. Plant Nutr. 2013, 36, 1496–1505. [CrossRef] Appl. Sci. 2019, 9, 4524 17 of 19 17 of 19 10. Malferrari, D.; Laurora, A.; Brigatti, F.; Coltorti, M.; Di Giuseppe, D.; Faccini, B.; Vezzalini, M. Open-field
experimentation of an innovative and integrated zeolitite cycle: Project definition and material characterization. Rend. Lincei-Sci. Fis. 2013, 24, 141–150. [CrossRef] 1. Colombani, N.; Di Giuseppe, D.; Faccini, B.; Ferretti, G.; Mastrocicco, M.; Coltorti, M. Estimated water sav
in an agricultural field amended with natural zeolites. Environ. Process. 2016, 3, 617–628. [CrossRef] 12. Di Giuseppe, D.; Ibáñez Insa, J.; Melchiorre, M.; Coltorti, M. On the potential effect of micronized zeolites on
seed germination: A prospective study. Period. Mineral. 2018, 85, 179–183. 13. Ferretti, G.; Di Giuseppe, D.; Faccini, B.; Coltorti, M. Mitigation of sodium risk in a sandy agricultural soil by
the use of natural zeolites. Environ. Monit. Assess. 2018, 190, 646. [CrossRef] [PubMed] 14. Reháková, M.; ˇCuvanová, S.; Dzivák, M.; Rimár, J.; Gaval’ová, Z. Agricultural and agrochemical uses of
natural zeolite of the clinoptilolite type. Curr. Opin. Solid State Mater. Sci. 2004, 8, 397–404. [CrossRef] 15. Ferretti, G.; Keiblinger, K.M.; Di Giuseppe, D.; Faccini, B.; Colombani, N.; Zechmeister-Boltenstern, S.;
Coltorti, M.; Mastrocicco, M. Short-term response of soil microbial biomass to different chabazite zeolite
amendments. Pedosphere 2018, 28, 277–287. 16. Bernardi, A.C.C.; Oliviera, P.P.A.; Monte, M.B.M.; Souza-Barros, F. Brazilian sedimentary zeolite use in
agriculture. Microporous Mesoporous Mater. 2013, 167, 16–21. [CrossRef] 17. Colombani, N.; Mastrocicco, M.; Di Giuseppe, D.; Faccini, B.; Coltorti, M. Variation of the hydraulic properties
and solute transport mechanisms in a silty-clay soil amended with natural zeolites. Catena 2014, 123, 195–204. [CrossRef] 18. Ferretti, G.; Keiblinger, K.M.; Zimmermann, M.; Di Giuseppe, D.; Faccini, B.; Colombani, N.; Mastrocicco, M.;
Zechmeister-Boltenstern, S.; Mentler, A. High resolution short-term investigation of soil CO2, N2O, NOx and
NH3 emissions after different chabazite zeolite amendments. Appl. Soil Ecol. 2017, 119, 138–144. [CrossRef]
19. Ferretti, G.; Di Giuseppe, D.; Natali, C.; Faccini, B.; Bianchini, G.; Coltorti, M. References International Soil Classification System for Naming
Soils and Creating Legends for Soil Maps; World Soil Resources Reports, No. 106; FAO: Rome, Italy, 2007. 28. Colombani, N.; Di Giuseppe, D.; Faccini, B.; Ferretti, G.; Mastrocicco, M.; Coltorti, M. Inferring the
interconnections between surface water bodies, tile-drains and an unconfined aquifer-aquitard system:
A case study. J. Hydrol. 2016, 537, 86–95. [CrossRef] 29. Orsini, L.; Rèmy, J.C. Utilization du chlorure de cobaltihexammine pour la déterminantion simultanée de la
capacité d′èchange et des bases échangeables des sols. Sci. Sol. 1976, 4, 269–275. Appl. Sci. 2019, 9, 4524 18 of 19 18 of 19 30. Ciesielsky, H.; Sterckeman, T. Determination of cation exchange capacity and exchangeable cations in soils
by means of cobalt heamine trichloride. Effects of experimental conditions. Agronomie 1997, 17, 1–7. 31. Mariotti, A. Natural N-15 abundance measurements and atmospheric nitrogen standards. Nature 1984, 311,
251–252. [CrossRef] 32. Silva, J.A.; Bremner, J.M. Determination and isotope-ratio analysis of different forms of nitrogen in soils:
5. Fixed ammonium. Soil Sci. Soc. Am. J. 1966, 30, 587–594. [CrossRef] 33. Ciavatta, C.; Govi, M.; Vittori Antisari, L.; Sequi, P. Characterization of humified compounds by extraction
and fractionation on solid polyvinylpyrrolidone. J. Chromatogr. 1990, 509, 41–146. [CrossRef] 34. Sequi, P.; De Nobili, M.; Leita, L.; Cercignani, G. A new index of humification. Agrochimica 1986, 30, 34. Sequi, P.; De Nobili, M.; Leita, L.; Cercignani, G. A new index of humification. Agrochimica 1986, 30, 175–179. 35. Vance, E.D.; Brookes, P.C.; Jenkinson, D.S. An extraction method for measuring soil microbial biomass C. Soil Biol. Biochem. 1987, 19, 703–707. [CrossRef] 35. Vance, E.D.; Brookes, P.C.; Jenkinson, D.S. An extraction method for measuring soil microbial biomass C. Soil Biol. Biochem. 1987, 19, 703–707. [CrossRef] 36. Ozbahce, A.; Tari, A.F.; Gönülal, E.; Simsekli, N.; Padem, H. The effect of zeolite applications on yield
components and nutrient uptake of common bean under water stress. Arch. Agron. Soil Sci. 2015, 61, 615–626. [CrossRef] 37. Barbarick, K.A.; Pirela, H.J. Agronomic and horticultural uses of zeolites: A review. In Zeo-Agriculture: Use of
Natural Zeolites in Agriculture and Aquaculture; Pond, W.G., Mumpton, F.A., Eds.; Westview Press: Boulder,
CO, USA, 1984; pp. 93–103. 38. Eslami, M.; Khorassani, R.; Coltorti, M.; Malferrari, D.; Faccini, B.; Ferretti, G.; Di Giuseppe, D.; Fotovat, A.;
Halajnia, A. Leaching behaviour of a sandy soil amended with natural and NH4+ and K+ saturated
clinoptilolite and chabazite. Arch. Agron. Soil Sci. 2018, 64, 1142–1151. [CrossRef] 39. References Dwairi, I.M. Evaluation of Jordanian zeolite tuffas a controlled slow-release fertilizer for NH4+. Environ. Geol. 1998, 34, 1–4. [CrossRef] 40. Dittert, K.; Georges, T.; Sattelmacher, B. Nitrogen turnover in soil after application of animal manure and
slurry as studied by the stable isotope 15N: A review. Z. Pflanzenernähr. Bodenkd. 1998, 161, 453–463. [CrossRef] 41. Högberg, P. Tansley review No. 95 15N natural abundance in soil-plant systems. N. Phytol. 1997, 137,
179–203. 2. Schmidt, O.; Ostle, N.J. Tracing nitrogen derived from slurry in earthworms using 15N/14N stable isot
ratios at natural abundances. Appl. Soil Ecol. 1999, 12, 7–13. [CrossRef] 43. Sang-Sun, L.; Woo-Jung, C.; Jyn-Hyeob, K.; Jae-Woon, J.; Scott, X.C.; Han-Yong, K.; Kwang-Sik, Y.;
Soo-Myung, C. Nitrogen and carbon isotope responses of Chinese cabbage and chrysanthemum to the
application of liquid pig manure. Plant Soil 2007, 295, 67–77. 44. Kitayama, K.; Iwamoto, K. Patterns of natural 15N abundance in the leaf-to-soil continuum of tropical rain
forests differing in N availability on Mount Kinabalu, Borneo. Plant Soil 2001, 229, 203–212. [CrossRef] 45. Gualtieri, A.F.;
Passaglia, E. Rietveld structure refinement of NH4-exchanged natural chabazite. Eur. J. Mineral. 2006, 18, 351–359. [CrossRef] 46. Hedström, A. Ion exchange of ammonium in zeolites: A literature review. J. Environ. Eng. 2001, 127, 673–681. [CrossRef] 47. Widiastuti, N.; Wu, H.; Ang, H.M.; Zhang, D. Removal of ammonium from greywater using natural zeolite. Desalination 2011, 277, 15–23. [CrossRef] 8. Malekian, R.; Abedi-Koupai, J.; Eslamian, S.S. Influences of clinoptilolite and surfactant-modified clinoptil
zeolite on nitrate leaching and plant growth. J. Hazard. Mater. 2011, 185, 970–976. [CrossRef] ,
;
p , J ;
,
p
p
zeolite on nitrate leaching and plant growth. J. Hazard. Mater. 2011, 185, 970–976. [CrossRef]
49. Azam, F.; Malik, K.A.; Hussain, F. Microbial biomass and mineralization-immobilization of nitrogen in some
agricultural soils. Biol. Fertil. Soils 1986, 2, 157–163. [CrossRef] 49. Azam, F.; Malik, K.A.; Hussain, F. Microbial biomass and mineralization-immobilization of nitrogen in some
agricultural soils. Biol. Fertil. Soils 1986, 2, 157–163. [CrossRef] 50. Ito, O.; Watanabe, I. Immobilization, mineralization and availability to rice plants of nitrogen derived from
heterotrophic nitrogen fixation in flooded soil. Soil Sci. Plant Nutr. 1981, 27, 169–176. [CrossRef] 51. Schulten, H.R.; Schnitzer, M. The chemistry of soil organic nitrogen: A review. Biol. Fertil. Soils 1998, 26,
1–15. [CrossRef] 52. Meurant, G. Soil Organic Matter and Its Role in Crop Production; Elsevier Science: Amsterdam, The Netherlands,
1973; p. 634. 53. References Scherer, H.W.; Werner, W. Significance of soil microorganisms for the mobilization of nonexchangeable
ammonium. Biol. Fertil. Soils 1996, 22, 248–251. [CrossRef] Appl. Sci. 2019, 9, 4524 19 of 19 54. Martens, D.A.;
Bremner, J.M. Urea hydrolysis in soils:
Factors influencing the effectiveness of
phenylphosphorodiamidate as a retardant. Soil Biol. Biochem. 1984, 16, 515–519. [CrossRef] 55. Li, C.; Fan, X.; Mengel, K. Turnover of interlayer ammonium in loess-derived soil grown with winter wheat
in the Shaanxi Province of China. Biol. Fertil. Soils 1990, 9, 211–214. [CrossRef] 56. Marzadori, C.; Vittori Antisari, L.; Gioacchini, P.; Sequi, P. Turnover of interlayer ammonium in soil cropped
with sugar beet. Biol. Fertil. Soils 1994, 18, 27–31. [CrossRef] 57. Hodge, A.; Robinson, D.; Fitter, A. Are microorganisms more effective than plants at competing for Nitrogen? Trends Plant Sci. 2000, 5, 304–308. [CrossRef] 58. Nannipieri, P.; Ascher, J.; Ceccherini, M.T.; Landi, L.; Pietramellara, G.; Renella, G. Microbial diversity and
soil functions. Eur. J. Soil Sci. 2003, 54, 655–670. [CrossRef] 59. Vries, F.T.; Hoffland, E.; Eekeren, N.; Brussaard, L.; Bloem, J. Fungal/bacterial ratios in grasslands with
contrasting nitrogen management. Soil Biol. Biochem. 2006, 38, 2092–2103. [CrossRef] 60. Strickland, M.S.; Rousk, J. Considering fungal:bacterial dominance in soils—Methods, controls, and ecosystem
implications. Soil Biol. Biochem. 2010, 42, 1385–1395. [CrossRef] 61. Merckx, R.; Van Ginkel, J.H.; Sinnaeve, J.; Cremers, A. Plant induced changes in the rhizosphere of maize and
wheat. I. Production and turnover of root-derived material in the rhizosphere of maize and wheat. Plant Soil
1986, 96, 85–93. [CrossRef] 62. Liljeroth, E.; Van Veen, J.A.; Miller, H.J. Assimilate translocation to the rhizosphere of two wheat cultivars
and subsequent utilization by rhizosphere microorganisms at two soil nitrogen levels. Soil Biol. Biochem. 1990, 22, 1015–1021. [CrossRef] 63. Capasso, S.; Salvestrini, S.; Coppola, E.; Buondonno, A.; Colella, C. Sorption of humic acid on zeolitic tuff:
A preliminary investigation. Appl. Clay Sci. 2005, 28, 159–165. [CrossRef] 64. Söderström, B.; Hedlund, K.; Jackson, L.E.; Kätterer, T.; Lugato, E.; Thomensen, I.K.; Jørgensen, H.B. What
are the effects of agricultural management on soil organic carbon (SOC) stocks? Environ. Evid. 2014, 3, 2. [CrossRef] 65. Evans, D.R. Physiological mechanism influencing plant nitrogen isotope composition. Trends Plant Sci. 2001,
6, 121–126. [CrossRef] 66. Evans, D.R.; Bloom, A.J.; Sukrapanna, S.S.; Ehleringer, J.R. Nitrogen isotope composition of tomato
(Lycopersicum Esculentum Mill. cv. T-5) grown under ammonium or nitrate nutrition. Plant Cell Environ. 1996, 19, 1317–1323. [CrossRef] 67. © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/). References Erskine, P.D.; Bergstrom, D.M.; Schmidt, S.; Stewart, G.R.; Tweedie, C.E.; Shaw, J.D. Subantarctic Macquarie
Island ± a model ecosystem for studying animal-derived nitrogen sources using 15N natural abundance. Oecologia 1998, 117, 187–193. [CrossRef] [PubMed] 68. Hulton, B.Z.; Sigman, D.M.; Schuur, E.A.G.; Edin, L.O. A climate-driven switch in plant nitrogen acquisition
within tropical forest communities. Proc. Natl. Acad. Sci. USA 2007, 114, 8902–8906. [CrossRef] [PubMed] 69. Kahmen, A.; Wanek, W.; Buchmann, N. Foliar d15N values characterize soil N cycling and reflect nitrate
or ammonium preference of plants along a temperate grassland gradient. Oecologia 2008, 156, 861–870. [CrossRef] [PubMed] 70. Choi, W.J.; Ro, H.M.; Hobbie, E.A. Patterns of natural 15N in soils and plants from chemically and organically
fertilized uplands. Soil Biol. Biochem. 2003, 35, 1493–1500. [CrossRef] 71. Bateman, A.S.; Kelly, S.D.; Jickells, T.D. Nitrogen Isotope Relationships between Crops and Fertilizer:
Implications for Using Nitrogen Isotope Analysis as an Indicator of Agricultural Regime. J. Agric. Food Chem. 2005, 53, 5760–5765. [CrossRef] 72. Yun, S.I.; Ro, H.M.; Choi, W.J.; Chang, S.X. Interactive effects of N fertilizer source and timing of fertilization
leave specific N isotopic signatures in Chinese cabbage and soil. Soil Biol. Biochem. 2006, 38, 1682–1689. [CrossRef] © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/).
|
https://openalex.org/W2550656211
|
https://orca.cardiff.ac.uk/id/eprint/96326/1/20160550.full.pdf
|
English
| null |
Contact probing of stretched membranes and adhesive interactions: graphene and other two-dimensional materials
|
Proceedings - Royal Society. Mathematical, physical and engineering sciences
| 2,016
|
cc-by
| 12,349
|
Subject Areas: nanotechnology, mathematical modelling,
mechanics Research Cite this article: Borodich FM, Galanov BA. 2016 Contact probing of stretched membranes
and adhesive interactions: graphene and other
two-dimensional materials. Proc.R.Soc.A
472: 20160550. Feodor M. Borodich1 and Boris A. Galanov2 1School of Engineering, Cardiff University, Cardiff CF24 0AA, UK
2Institute for Problems in Materials Science, National Academy of
Sciences of Ukraine, 3 Krzhyzhanovsky Street, Kiev 03142, Ukraine
FMB, 0000-0002-7935-0956 http://dx.doi.org/10.1098/rspa.2016.0550 Received: 9 July 2016
Accepted: 17 October 2016 Contact probing is the preferable method for studying
mechanical properties of thin two-dimensional (2D)
materials. These studies are based on analysis of
experimental force–displacement curves obtained by
loading of a stretched membrane by a probe of an
atomic force microscope or a nanoindenter. Both non-
adhesive and adhesive contact interactions between
such a probe and a 2D membrane are studied. As
an example of the 2D materials, we consider a
graphene crystal monolayer whose discrete structure
is modelled as a 2D isotropic elastic membrane. Initially,
for
contact
between
a
punch
and
the
stretched circular membrane, we formulate and solve
problems that are analogies to the Hertz-type and
Boussinesq frictionless contact problems. A general
statement for the slope of the force–displacement
curve is formulated and proved. Then analogies
to the JKR (Johnson, Kendall and Roberts) and
the Boussinesq–Kendall contact problems in the
presence of adhesive interactions are formulated. General nonlinear relations among the actual force,
displacements and contact radius between a sticky
membrane and an arbitrary axisymmetric indenter
are derived. The dimensionless form of the equations
for power-law shaped indenters has been analysed,
and the explicit expressions are derived for the
values of the pull-off force and corresponding critical
contact radius. on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from Contact probing of stretched
membranes and adhesive
interactions: graphene and
other two-dimensional
materials
Feodor M. Borodich1 and Boris A. Galanov2
on November 29, 2016
alsocietypublishing.org/ rspa.royalsocietypublishing.org rspa.royalsocietypublishing.org Keywords: two-dimensional materials, graphene,
membrane, Johnson, Kendall and Roberts
theory, adhesion, nanoindentation 1. Introduction 2 rspa.royalsocietypublishing.org Proc. R.Soc. A 472: 20160550
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . For many years, it was assumed that atomically thin two-dimensional (2D) materials could not
exist in the free state due to their instability (e.g. a discussion by Meyer et al. [1]). The group
led by Geim showed that layered materials may be exfoliated using a sticky tape; in particular,
one can produce atomically thin 2D carbon allotrope, named graphene, and that graphene is
stable [1,2]. The group led by Ruoff showed that there are other methods for producing graphene-
based materials (like graphene oxide) in significant quantities [3,4]. Currently, 2D materials are a
wide new field in condensed matter research [5]. In addition to graphene and graphene oxide
these 2D materials include mica, weakly coupled planes of CuO2, MoS2, NbSe2 WSe2, TaS2,
TaSe2, etc. [5–7]. These 2D materials have a great potential for applications in modern electronics. Indeed, it is known that graphene has remarkable electronic properties, high tensile strength and
low density (e.g. [8,9]), hence it is considered as an excellent material for nanoelectromechanical
systems (NEMSs). Examples of these systems include the circular drum resonators covered
by graphene, pressure sensors and membrane-based NEMS devices with suspended graphene
spanned between electrodes (e.g. [10–12]). Nowadays studies of 2D materials are concentrated not only on their remarkable electronic
properties but also on their mechanical behaviour. Contact probing is the preferable method for
studying mechanical properties of thin 2D materials (e.g. [4,13–15]). A comprehensive review
of 2D materials, methods of their fabrications and studies of their mechanical properties have
been recently published by Castellanos-Gomez et al. [7]. Usually, the probe of an atomic force
microscope (AFM) or a nanoindenter is used. If a probe is applied at the centre of a suspended
2D material, i.e. at the centre of a circular membrane or in the middle of a doubly clamped
beam, then one can record the external load (P) and the corresponding displacement (δ) of
the indenter in order to get the force–displacement curve. This is the so-called depth-sensing
nanoindentation. on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from Author for correspondence: 2016 The Authors. Published by the Royal Society under the terms of the
Creative Commons Attribution License http://creativecommons.org/licenses/
by/4.0/, which permits unrestricted use, provided the original author and
source are credited. rspa.royalsocietypublishing.org Proc. R.Soc. A 472: 20160550
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Thus, for proper understanding of interactions between the tip and a 2D membrane, one needs
to study the influence of the adhesive forces on their contact. Graphene may be considered as one
of the main classes of 2D materials. Although adhesive contact problems for elastic solids and
elastic membranes have been studied for a long period of time (e.g. [29,30] and reference therein)
and the contact probing of graphene membranes is a very popular technique (e.g. [7]) to the best
of our knowledge, the adhesive contact problem for atomically thin object (e.g. graphene layer)
has not been studied yet. In addition, the used theoretical models assumed non-zero bending
rigidity of the membranes. In this paper, a 2D layer under tensile loading is modelled as a linear
elastic membrane whose bending rigidity may be neglected. The membrane is isotropic because
it is known [31] that the elastic in-plane properties of 2D graphene layer in a linear approximation
are isotropic. To solve contact problems with molecular adhesion for a stretched membrane, the
JKR approach along with Derjaguin’s ideas will be employed. The former approach involves
calculations of the work done by the surface attractions and the work of deformation in the elastic
objects [22,25]. The paper is organized as follows: In §2, we present some preliminary information about graphene and some other carbon-
based materials. Then we discuss formulations of the classic frictionless non-adhesive Hertz and
Boussinesq contact problems and mechanics of adhesive contact for elastic three-dimensional
(3D) solids. In §3, we first discuss equations of elastic stretched membranes. Then for the stretched 2D
membranes, we formulate the non-adhesive frictionless contact problems that are analogies to
the corresponding Hertz and Boussinesq contact problems. These new problems are solved using
the Green function approach. For the problems under consideration, a general statement about
slopes of the force–displacement relations is formulated and proved. The JKR theory of adhesive contact was originally developed for linearly elastic isotropic
spheres. In §4, the adhesive contact between a stretched graphene membrane and an
axisymmetric convex punch of arbitrary profile is formulated and solved in the framework of
the JKR theory. The derivation of the main relations is quite straightforward because it is based
on the use of the above mentioned general statement. 2. Preliminaries We use both the Cartesian and cylindrical coordinate frames, namely x1 = x, x2 = y, x3 = z and
r, ϕ, z, where r =
x2 + y2 and x = r cos ϕ, y = r sin ϕ. rspa.royalsocietypublishing.org Proc. R.Soc. A 472: 20160550
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Then connections between the obtained
results and problems of adhesive probing of graphenes are discussed assuming that the indenter
shape is described by a power-law function of an arbitrary real degree m > 1. In this case, the
exact relations between the actual force, the indenter displacements and contact radius have been
derived and the explicit expressions are found for the values of the pull-off force and for the
corresponding critical contact radius. Special attention is given to problems for spherical and
flat-ended circular punches. 1. Introduction Using analysis of the P–δ curve, one can try to extract the mechanical properties
of the material. Usually, the classic P–δ expressions [16] for doubly clamped structures have
been used [7]. However, these expressions are valid for structures of finite thickness of non-zero
bending rigidity. There is a need for studying contact problems for atomically thin 2D materials. In nanoindentation tests of 2D materials, deformations of a freestanding membrane may
depend not only on the force P acting on the probe but also on the attraction between the
probe tip and the material due to interaction forces having electromagnetic origin. Discussing
the phenomenon of adhesion, Robert Hooke wrote ‘for the Congruity, in the Vibrative motions,
may be the cause of all kind of attraction, not only Electrical, but Magnetical also, and therefore it
may be also of Tenacity and Glutinousness.’ [17]. Peter Lebedev [18] gave the first electromagnetic
explanation for the nature of the van der Waals (vdW) forces. Later it was realized that the vdW
forces include forces of different origins (the Keesom, Debye and London forces) that mean,
respectively, attraction between: two permanent dipoles, a permanent dipole and a corresponding
induced dipole and two instantaneously induced dipoles [19,20]. Various ways for description of
vdW interactions and adhesion between solids are still discussed by the scientific community. Recently, it has been argued that a qualitatively correct description of the vdW interactions
between polarizable nanostructures over a wide range of finite distances can only be attained
by accounting for the wavelike nature of charge density fluctuations [21]. Molecular adhesion
becomes increasingly significant as the contact size decreases [20], hence, one can expect that
adhesive interactions may have a great influence on the contact between a probe and a sample of
a 2D material. There are various approaches to adhesive contact problems. In particular, these approaches
may include the use of the Derjaguin approximation and the energy approach [22] and the use
of an appropriate interaction potential between points on the surfaces, for example, a Lennard–
Jones potential or piecewise-constant approximations of these potentials [23,24]. Nowadays, there
are several classic approaches to describe adhesive contact between solid spheres that include the
JKR (Johnson, Kendall and Roberts) and the DMT (Derjaguin–Muller–Toporov) theories that may
be considered as limits of the Maugis transition between the JKR and DMT approaches [23,25,26]. on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from To study contact problems with molecular adhesion one needs to know the work of adhesion,
w that is equal to the energy needed to separate two dissimilar surfaces from contact to infinity. The above classic models are very helpful for studying various phenomena that involve molecular
adhesion. For example, the non-direct BG method for the experimental determination of the work
of adhesion and elastic contact modulus of materials [27] is based on the use of these classic
models. It has been shown recently that this non-direct method is fast and robust [28]. The similar
techniques should be developed for problems of adhesive contact between an indenter and a
stretched membrane of 2D material. 3 rspa.royalsocietypublishing.org Proc. R.Soc. A 472: 20160550
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The carbon atoms in the graphene crystal are bonded to each other by covalent bonds. It is
clear that a linearization of interatomic interactions will represent the covalent bonds as an elastic
spring whose elastic constant is one of the discrete model parameters. A comprehensive review of
existing approaches to modelling the elastic in-plane properties of graphene has been presented
by Berinskii & Borodich [41]. It was shown that a number of popular linear and nonlinear models
of graphene lattices (in particular, popular nonlinear base on the use of the Tersoff or Brenner
potentials) have serious flaws and often the results obtained using these models do not have
physical meaning. On the other hand, the two-parametric molecular mechanics model developed
by Gillis [42] where parameters represent the central and non-central interaction of carbon
atoms, can well describe the experimental results for small in-plane deformations of graphene. Nowadays, it is still not clear what is better to consider as thickness of the graphene layer and
to avoid this problem, the elastic modulus for graphene and nanotubes are often defined as a
product of the conventional Young’s modulus with the layer thickness (see a discussion in [41]). The rigorous studies of the graphene crystal lattice as 2D linear elastic continuum [31] showed
that the lattice has isotropic elastic properties. p
p
p
The graphene oxide (G–O) is another 2D carbon-based material. It is a graphene plane
contaminated by sp3-hybridized carbons bearing hydroxyl and epoxide functional groups on the
plane, and by carboxyl and carbonyl groups at the plane edges. As it was noted by Suk et al. [4],
the detailed understanding of G–O structure is still being developed. One may assume that the
main arguments presented by Berinskii & Borodich [31] in application to pure graphene, are also
valid in application to G–O and therefore, this 2D material has also isotropic elastic properties. (a) Graphene and other carbon-based materials Graphene may be considered as the main example of 2D materials. The term graphene denotes
a carbon allotrope that can be described as an atom thick sheet made of its atoms. In fact, on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from graphene is a single layer of graphite, i.e. the atoms of graphene are densely packed into a 2D
honeycomb crystal lattice. Owing to symmetry of the graphene lattice, a hexagonal ring may
be used to represent its structure. The initial length of the hexagon side (carbon interatomic
distance) is a = 1.42 Å and the initial angle between two any sides is α = 120◦. Properties of
carbon allotropes were studied for many years (e.g. reviews by Derjaguin & Fedoseev [32]). Initially, these studies were concentrated on graphite and diamond allotropes. Tubular carbon
allotropes that later started to be called carbon nanotubes (CNTs) were discovered only in 1950
due to the electron microscopy studies fulfilled by Radushkevich & Lukyanovich [33,34] in the
Institute of Physical Chemistry (USSR Academy of Sciences) where an extended programme on
carbon research was led by Derjaguin (Department of Surface Forces) and Dubinin (Department
of Sorption Processes). The results of diffraction studies of the carbon hollow whiskers (CNTs)
showed that CNTs are made from the pure graphene planes elongated along the tubes and this can
explain high tensile strength of the CNTs [35]. Thus, graphene is a basic building block for many
graphitic materials of other dimensionalities [36]. The mechanical properties of graphene have
been experimentally studied using various approaches (e.g. [10,37,38]). On the theoretical front
the elastic in-plane properties of graphene were studied using various linearized discrete models
and models involving various nonlinear multi-body potentials (e.g. discussions in [39,40]). 4 (c) The adhesive contact models Derjaguin [22] presented the first attempt to consider the problem of adhesion between elastic
spheres or between an elastic sphere and an elastic half-space. He assumed that the deformed
shape of the sphere can be calculated by solving the Hertz contact problem (this assumption was
not correct) and suggested to calculate the adhesive interaction using the so-called Derjaguin
approximation (this approximation is very useful). This approximation does not employ the
pairwise summation of the interactions between all elements of solids, but it reduces the volume
molecular attractions to surface interactions. The Derjaguin idea that the virtual work done by the
external load is equal to the sum of the virtual change of the potential elastic energy and the virtual
work that will be consumed by the increase of the surface attractions (see (21) in Derjaguin [22]),
is the cornerstone of many approaches to adhesive contact problems. For example, Sperling [46]
used this idea to calculate the full energy of the system and derived the equations that are identical
to equations of the JKR theory (see a discussion in [44] for details). However, the JKR approach
is more elegant. Indeed, to get the final result, Sperling [46] solved the problem using rather
complicated calculations, while according to the JKR approach, the problem may be solved by
assuming that the contact system has come to its real state in two steps: (i) first it has got real
(true) contact radius a1 and an apparent depth of indentation δ1 under some apparent Hertz
load P1, then (ii) using the Boussinesq solution for contact a flat-ended punch of radius a1, the
system is unloaded from P1 to a real value of the external load P0 (the true external load) keeping
the contact radius a1 constant. The original JKR approach has the same assumptions as the
frictionless axisymmetric Hertz contact. Although the approach was applied to a sphere described
as a paraboloid of revolution z = r2/(2R), very far generalizations of the approach are possible. Recently, the approach has been extended to non-slipping boundary conditions [47], transversely
isotropic solids [48] and the punches of arbitrary axisymmetric blunt shapes contacting elastic
materials with rotational symmetry of their elastic properties [44]. on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from is equal to the initial distance between the surfaces. Hence, the equation of the punch surface
given by a function f, can be written as x3 = −f(x1, x2), f ≥0. After the punch contacts with the
half–space, displacements ui and stresses σij are generated. is equal to the initial distance between the surfaces. Hence, the equation of the punch surface
given by a function f, can be written as x3 = −f(x1, x2), f ≥0. After the punch contacts with the
half–space, displacements ui and stresses σij are generated. 5 rspa.royalsocietypublishing.org Proc. R.Soc. A 472: 20160550
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . j
Using a known expression for a potential of an ellipsoid, the contact problem for solids whose
shapes are approximated as elliptic paraboloids, was solved by Hertz [43]. Hertz showed also that
the contact region of the problem is an ellipse. Boussinesq [45] presented independently solutions
to other contact problems. The Boussinesq contact problem for a flat-ended cylinder assumes that
the contact region is always a circle of a fixed radius. (b) The Hertz and Boussinesq contact problems for an elastic-half-space The non-adhesive 3D Hertz formulation assumes that initially there is only one point of contact
between two elastic solids. In a geometrically linear formulation of the boundary-value contact
problem, each solid is modelled as a positive half-space x3 ≥0. The boundary plane x3 = 0 is
denoted by R2. The origin (O) of Cartesian x1, x2, x3 coordinates is at the point of initial contact
between the solids. Assuming that contact is frictionless and that the radial displacements may
be neglected, Hertz [43] gave a formulation of the boundary-value problem (see for details [44]). In particular, the Hertz type of contact problems means that the contact region is unknown in
advance, and only vertical displacements of the boundary are taken into account. Hence, one
needs to find the finite region D of the points at which the bodies are in mutual contact, the
relative approach of the bodies δ > 0 and the displacements and the stresses within the solids. It is possible to show that the problem formulation is mathematically equivalent to the problem
of contact between a positive half-space and a rigid indenter (punch) whose shape function f 3. Contact problems for a circular graphene membrane: analogies to the Hertz
and Boussinesq problems Owing to specific character of 2D materials, it is reasonable to model a material layer as a
membrane whose bending rigidity is negligible. Modelling the material as an elastic membrane
allows us to use a geometrically linear formulation of the boundary-value problem like the classic
contact problems have. Of course, the formulation has to be slightly modified. We consider only
axisymmetric problems of frictionless contact between punches described as bodies of revolution
and a circular stretched membrane (a circular drum). Evidently, the above Hertz and Boussinesq
contact problems formulated for an elastic-half-space have to be reformulated to reflect the
features of a 2D solid. (a) The elastic membrane equation and the Green function solution Mathematical problems related to stretched membranes have been studied by many researchers
(e.g. [49,50]). Hence, we provide here just formulations of the appropriate boundary-value
problem. Let us consider an elastic membrane Ω that is supported by a rigid frame in
the horizontal plane R2. Let the membrane be held under a uniform tension T, i.e. initially the
membrane is stretched. Hence, each point (x, y) of the closure ¯Ω of Ω represents a material point
of the membrane when it is stretched without any other applied force. If some external force of density F(x, y) is applied perpendicularly to the membrane surface
then vertical displacements of the membrane u3(x, y) appear. It is assumed that the internal forces
lay in the tangent plane to the deformed surface. Thus, the shape of the deformed surface at
equilibrium is presented as (x, y, u3(x, y)) and the tension T does not depend on u3(x, y), i.e. T is constant. The function u3(x, y) satisfies the elastic membrane equation that actually is the
following Poisson equation T∇2u3(x, y) = −F(x, y),
(x, y) ∈Ω,
∇2 = 1
r
∂
∂r
r ∂
∂r
+ 1
r2
∂2
∂ϕ2 = ∂2
∂x2 + ∂2
∂y2 ,
(3.1) (3.1) where ∇2 is the Laplace operator. If it is assumed additionally that the stretched membrane sticks to the border of the frame ∂Ω,
then one has a homogeneous Dirichlet boundary condition If it is assumed additionally that the stretched membrane sticks to the border of the frame ∂Ω,
then one has a homogeneous Dirichlet boundary condition u3(x, y) = 0,
(x, y) ∈∂Ω. (3.2) (3.2) Let us consider a circular drum, i.e. a circular membrane that is supported by a rigid circular
frame of radius R. (c) The adhesive contact models It is attempted here to follow
the original JKR approach as closely as possible, hence we denote as a1, P0 and δ2 the true values
of the radius of adhesive contact, the external load and the displacement, respectively. The total energy UT is obtained by summation of the stored elastic energy UE, the mechanical
energy in the applied load UM and the surface energy US UT = UE + UM + US. (2.1) (2.1) According to the Derjaguin approximation and the JKR assumption that the adhesive interactions
out of the contact region are negligible, the surface energy can be written as According to the Derjaguin approximation and the JKR assumption that the adhesive interactions
out of the contact region are negligible, the surface energy can be written as US = −wA,
(2.2) (2.2) where A is the area of the contact region. where A is the area of the contact region. Sperling [46] argued that the total energy has minimum at equilibrium, whereas Johnson
et al. [25] argued that the energy satisfies the Griffith criterion. However, both arguments lead
to the same equation dUT
da1
= 0
or
dUT
dP1
= 0. (2.3) (2.3) It is known that the frictionless JKR results can be also obtained by linear fracture mechanics
approach [23]. If G is the energy release rate at the edge of the contact then in the frictionless on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from case, the equilibrium is given by G = w (Griffith’s criterion). However, the concepts of fracture
mechanics cannot be directly applied in the case of atomically thin 2D membranes. Thus, the
energy approach will be used further. case, the equilibrium is given by G = w (Griffith’s criterion). However, the concepts of fracture
mechanics cannot be directly applied in the case of atomically thin 2D membranes. Thus, the
energy approach will be used further. 6 (a) The elastic membrane equation and the Green function solution A schematic of contact between a convex smooth axisymmetric punch f(r) and an elastic membrane. Dash lines
correspond to the problem without an external load (P = 0) and the solid lines describe the system state after application of
the load P. Here T is the tension in the membrane; R and a are the radii of the drum and the contact region, respectively, and δ
is the displacement of the punch nose. acting on a sector of the membrane with the central angle ϕ is equal to Tr ϕ du3/dr. Therefore,
the equation of equilibrium of the membrane along the vertical axis z under a vertical force of
intensity F(r) per unit area is d
dr
Tr ϕ du3
dr
+ F(r)r ϕ = 0. In this case, the equation (3.1) and the boundary condition (3.2) can be written as In this case, the equation (3.1) and the boundary condition (3.2) can be written as T 1
r
d
dr
rdu3
dr
+ F(r) = 0,
0 ≤r ≤R
(3.5) (3.5) and u3(r) = 0,
r = R. (3.6) (3.6) (a) The elastic membrane equation and the Green function solution The Green function G(r, ϕ, ρ, ψ) for a circular membrane having the Dirichlet
boundary condition can be written as [51] G(r, ϕ, ρ, ψ) = 1
4π ln R2 + r2ρ2/R2 −2rρ cos(ϕ −ψ)
r2 + ρ2 −2rρ cos(ϕ −ψ)
(3.3) (3.3) and the solution u3(r, ϕ) to the elastic membrane equation (3.1) with homogeneous boundary
condition (3.2) is presented in the form and the solution u3(r, ϕ) to the elastic membrane equation (3.1) with homogeneous boundary
condition (3.2) is presented in the form u3(r, ϕ) = 1
T
Ω
G(r, ϕ, ρ, ψ)F(ρ, ψ)ρ dρ dψ = 1
T
2π
0
R
0
G(r, ϕ, ρ, ψ)F(ρ, ψ)ρ dρ dψ. (3.4) (3.4) In the case of axial symmetry, the membrane deflection is only a function of the radius r and
it does not depend on the angle ϕ, i.e. u3(r, ϕ) ≡u3(r) . Hence, the partial derivative ∂/∂r can be
replaced by the total derivative d/dr. The elastic membrane equation (3.1) for this particular case
could be derived in a very simple way (e.g. [49]). Indeed, the vertical component of the tension on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from p
y
yp
g g
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . R
T
R
f
P
0
T
r
a
a
d
z
Figure 1. A schematic of contact between a convex smooth axisymmetric punch f(r) and an elastic membrane. Dash lines
correspond to the problem without an external load (P = 0) and the solid lines describe the system state after application of
the load P. Here T is the tension in the membrane; R and a are the radii of the drum and the contact region, respectively, and δ
is the displacement of the punch nose. R
T
R
f
P
0
T
r
a
a
d
z 7
rspa.royalsocietypublishing.org Proc. R.Soc. A 472: 20160550
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 Figure 1. on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from Note that, in the general case of an axisymmetric convex smooth punch f(r), f(0) = 0 acting on
the membrane, one has δ = u3(0, 0). It follows from (3.3) that 8 8
rspa.royalsocietypublishing.org Proc. R.Soc. A 472: 20160550
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . G(0, 0, ρ, ψ) = 1
4π ln R2
ρ2 . Substituting the above expression and (3.8) into (3.4), one obtains bove expression and (3.8) into (3.4), one obtains δ = u3(0, 0) = 1
T
2π
0
a
0
1
4π ln R2
ρ2 · T 1
ρ
d
dρ
ρ df(ρ)
dρ
ρ dρ dψ
(3.11) or or δ = 2π
4π
a
0
ln
R
ρ
2 d
dρ
ρ df
dρ
dρ = af ′(a) ln
R
a
+ f(a). (3.12) (3.12) If the punch shape is described by monomial (power-law) function of radius of an arbitrary
real degree m ≥1 If the punch shape is described by monomial (power-law) function of radius of an arbitrary
real degree m ≥1 f(r) = Bmrm,
(3.13) f(r) = Bmrm, (3.13) where m is the degree of the monomial function and Bm is the constant of the shape, then one has where m is the degree of the monomial function and Bm is the constant of the shape, then one has p(r) =
TBmm2rm−2
for r ≤a
0
for r > a. (3.14) (3.14) Hence, it follows from (3.10) that a =
P
2πmTBm
1/m
. (3.15) For a power-law punch (3.13), it follows from (3.11) that For a power-law punch (3.13), it follows from (3.11) that δ = Bmam
ln
R
a
m
+ 1
=
P
2πmT
1 −ln
P
2πmTBmRm
. (3.16) (3.16) Let us derive a general relation for slopes of δ–P curves similar to the relation derived
by Borodich [44] in the case of frictionless Hertz-type contact problem. The following general
statement for the slope of the force–displacement curve can be formulated and proved. (b) An analogy to the Hertz-type contact problem (b) An analogy to the Hertz-type contact problem If one considers an analogy to the axisymmetric Hertz-type contact problem then for a convex
smooth punch f(r) pressed by a vertical load P, one needs to find the radius a of unknown region
DC of contact between the punch and the graphene membrane and the displacement of the punch
nose δ (figure 1). It is known that within the contact region the displacements are given by u3(r) = δ −f(r),
r ≤a. (3.7) (3.7) In the general case, the contact pressure p(r) is acting along z axis, i.e. it creates positive force
density In the general case, the contact pressure p(r) is acting along z axis, i.e. it creates positive force
density p(r) = F(r) = −T 1
r
d
dr
rdu3
dr
= T 1
r
d
dr
rdf(r)
dr
,
r ≤a
(3.8) (3.8) because u′
3(r) = −f ′(r) for r ≤a; and p(r) = 0 for r > a. Note the pressure may be negative due to
the presence of adhesive interactions. The total vertical load acting on the membrane P satisfies the equation 2π
0
R
0
F(r)r dr dϕ = 2π
R
0
F(r)r dr = P. (3.9) (3.9) Hence, for an axisymmetric convex smooth punch f(r), f(0) = 0 acting on the membrane, one has Hence, for an axisymmetric convex smooth punch f(r), f(0) = 0 acting on the membrane, one has P = 2πT
a
0
d
dr
rdf(r)
dr
dr = 2πTaf ′(a). (3.10) (3.10) Here we have used the Newton fluxion notation f ′ to denote a derivative of f. on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from Let us derive a general relation for slopes of δ–P curves similar to the relation derived
by Borodich [44] in the case of frictionless Hertz-type contact problem. The following general
statement for the slope of the force–displacement curve can be formulated and proved. Let a circular linear elastic isotropic membrane be supported by a rigid circular frame of radius
R and the membrane be stretched by a uniform tension T. Let the behaviour of the membrane be
described by (3.5) and (3.6). Let an axisymmetric convex, smooth (f ∈C2(R2 \ {0})) punch f(r),
f(0) = 0 be in contact with the membrane under action of the load P and the contact problem be
described by (3.7)–(3.9). y
Then the slope of the δ–P curve at any point is Then the slope of the δ–P curve at any point is dP
dδ = K,
K =
2πT
ln(R/a),
(3.17) (3.17) he radius of the contact region. where a is the radius of the contact region. where a is the radius of the contact region. Indeed, it follows from (3.10) that P = 2πTaf ′(a). Then one has Indeed, it follows from (3.10) that P = 2πTaf ′(a). Then one has dP
da = 2πT[f ′(a) + af ′′(a)]. (3.18) (3.18) he other hand, it follows from (3.12) that δ = [af ′(a) ln(R/a) + f(a)] and, therefore, On the other hand, it follows from (3.12) that δ = [af ′(a) ln(R/a) + f(a)] and, therefore, On the other hand, it follows from (3.12) that δ = [af (a) ln(R/a) + f(a)] and, therefore,
dδ
da =
f ′(a) ln
R
a
+ af ′′(a) ln
R
a
−af ′(a)1
a + f ′(a)
= ln
R
a
[f ′(a) + af ′′(a)]. (3.19) dδ
da =
f ′(a) ln
R
a
+ af ′′(a) ln
R
a
−af ′(a)1
a + f ′(a)
= ln
R
a
[f ′(a) + af ′′(a)]. Comparing (3.18) and (3.19), one obtains Comparing (3.18) and (3.19), one obtains dP
da = K dδ
da dP
da = K dδ
da that leads to (3.17). that leads to (3.17). on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from R
T
R
P
0
T
r
a
a
d
z
Figure 2. on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from A schematic of contact between a flat-ended axisymmetric punch and an elastic membrane. Dash lines correspond
to the position of the membrane without action of an external load (P = 0) and the solid lines describe the system state after
application of the load P. Here T is the tension in the membrane; R and a are the radii of the drum and the punch, respectively,
and δ is the displacement of the punch. 9 9
rspa.royalsocietypublishing.org Proc. R.Soc. A 472: 20160550
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Figure 2. A schematic of contact between a flat-ended axisymmetric punch and an elastic membrane. Dash lines correspond
to the position of the membrane without action of an external load (P = 0) and the solid lines describe the system state after
application of the load P. Here T is the tension in the membrane; R and a are the radii of the drum and the punch, respectively,
and δ is the displacement of the punch. (c) An analogy to the Boussinesq contact problem If one considers an analogy to the axisymmetric Boussinesq contact problem for a flat-ended
punch of radius a pressed by a vertical load P (figure 2), then it follows from (3.5) that the pressure
p(r) = 0 for 0 ≤r < a. Hence, one has p(r) = Cδ(r −a), where δ(r −a) is the Dirac delta-function having its support on the circle of radius a and the
constant C is defined from the condition that the total vertical load acting on the membrane P
satisfies the equation P =
2π
0
R
0
p(r)r dr dϕ =
2π
0
R
0
Cδ(r −a)r dr dϕ = 2πaC. (3.20) (3.20) Hence, C = P/(2πa). Owing to axial symmetry of the problem, u3(r, ϕ) = u3(r, 0) = u3(r), i.e. one can put ϕ = 0 in the
expression for the Green function and correspondingly into (3.4) Owing to axial symmetry of the problem, u3(r, ϕ) = u3(r, 0) = u3(r), i.e. one can put ϕ = 0 in the
expression for the Green function and correspondingly into (3.4) u3(r) = C
T
1
4π
2π
0
R
0
ln R2 + r2ρ2/R2 −2rρ cos ψ
r2 + ρ2 −2rρ cos ψ
δ(ρ −a)ρ dρ dψ or or u3(r) = Ca
T
1
4π
2π
0
ln R2 + r2a2/R2 −2ra cos ψ
r2 + a2 −2ra cos ψ
dψ. (3.21) (3.21) Using a known expression (e.g. 865.73 at [52]) Using a known expression (e.g. 865.73 at [52]) 2π
0
ln(k2 ± 2kl cos ψ + l2) dψ = 4π ln k,
for 0 ≤l ≤k,
(3.22) 2π
0
ln(k2 ± 2kl cos ψ + l2) dψ = 4π ln k,
for 0 ≤l ≤k,
(3.22) (3.22) one can calculate the membrane deviation by rearranging the above integral (3.21), into the
following form u3(r) = Ca
T
1
4π
2π
0
ln R2/(ra) + ra/R2 −2 cos ψ
r/a + a/r −2 cos ψ
dψ. (3.23) (3.23) Let us consider the membrane deflection outside the contact region, i.e. the case a ≤r ≤R. Putting
k2 = R2/(ra) and l2 = ra/R2, and applying (3.22) to the nominator of (3.23) and then putting k2 = r/a
and l2 = a/r and applying (3.22) to the denominator, one gets Let us consider the membrane deflection outside the contact region, i.e. (c) An analogy to the Boussinesq contact problem the case a ≤r ≤R. Putting
k2 = R2/(ra) and l2 = ra/R2, and applying (3.22) to the nominator of (3.23) and then putting k2 = r/a
and l2 = a/r and applying (3.22) to the denominator, one gets u3(r) = Ca
T
1
4π 4π
ln
R
√ra −ln
r
a
= aC
T ln R
r =
P
2πT ln R
r . (3.24) (3.24) Let us consider now the membrane deflection inside the contact region, i.e. the case 0 ≤r ≤a. Putting k2 = R2/(ra) and l2 = ra/R2, and applying (3.22) to the nominator of (3.21) and then putting on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from l2 = r/a and applying (3.22) to the denominator, one gets k2 = a/r and l2 = r/a and applying (3.22) to the denominator, one gets 10 u3(r) = Ca
T
1
4π 4π
ln
R
√ra −ln
a
r
= aC
T ln R
a =
P
2πT ln R
a = const. (3.25) rspa.royalsocietypublishing.org Proc. R.Soc. A 472: 20160550 (3.25) The logarithmic decay in the membrane solution is in agreement with known solutions that were
obtained for membranes of finite thickness (e.g. [29,30]). Thus, it follows from (3.25) that the displacement δ of a flat-ended punch of radius a acting
on an elastic membrane of radius R and stretched by the uniform tension T (figure 2), is a linear
function of the external load P δ = u3(r) = P
K ,
K =
2πT
ln(R/a), 0 ≤r ≤a. (3.26) (3.26) The formulae (3.24) and (3.25) give the full solution to the contact problem for a circular flat-ended
punch. The formulae (3.24) and (3.25) give the full solution to the contact problem for a circular flat-ended
punch. 4. Adhesion to a circular graphene monolayer membrane 4. Adhesion to a circular graphene monolayer membrane Here the JKR theory will be extended to a circular graphene monolayer membrane. The original
theory was developed for adhesive contact between two isotropic elastic spheres. on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from The total energy UT is calculated according to (2.1). Therefore, one has energy UT is calculated according to (2.1). Therefore, one has 11 11
rspa.royalsocietypublishing.org Proc. R.Soc. A 472: 20160550
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . UT = P1δ1 −
P1
0
δ(P) dP −(P2
1 −P2
0)
2K(a1)
−P0δ1 + P0
(P1 −P0)
K(a1)
−wπa2
1. Taking into account that Taking into account that −(P2
1 −P2
0)
2K(a1)
+ P0
(P1 −P0)
K(a1)
= −(P1 −P0)2
2K(a1)
, one has UT = (P1 −P0)δ1 −
P1
0
δ(P) dP −(P1 −P0)2
2K(a1)
−wπa2
1. (4.6) (4.6) This expression is an analogy to the expression obtained for a generalized JKR contact theory in
the case of 3D elasticity [44]. Because the equilibrium state satisfies the equation (2.3), we need to calculate the appropriate
derivatives. First, let us get the following expressions d
dP1
[(P1 −P0)δ1] = (P1 −P0) dδ1
dP1
+ δ1,
d
dP1
P1
0
δ(P) dP = δ(P1) = δ1,
d
dP1
(P1 −P0)2
2K(a1)
= (P1 −P0)
K(a1)
−(P1 −P0)2
2
1
2πTa1
da1
dP1
. It has been taken into account in the above expression that d
dP1
1
K(a1)
=
d
dP1
ln(R/a1)
2πT
= −
1
2πTa1
da1
dP1
. Hence, we get Hence, we get dUT
dP1
= (P1 −P0) dδ1
dP1
−(P1 −P0)
K(a1)
+
(P1 −P0)2
4a1πT
−2wπa1
da1
dP1
= 0. (4.7) (4.7) Using (3.17) of the above general statement, one obtains from (4.7) that the equilibrium condition
for the general JKR model is Using (3.17) of the above general statement, one obtains from (4.7) that the equilibrium condition
for the general JKR model is dUT
dP1
=
(P1 −P0)2
4a1πT
−2wπa1
da1
dP1
= 0
(4.8) (4.8) or (P1 −P0)2 = 8π2wTa2
1. (a) General expressions for an arbitrary axisymmetric punch As one neglects the adhesive forces acting outside the contact region, it follows from (2.2) that the
surface energy can be written as
2 US = −wπa2
1. (4.1) US = −wπa2
1. (4.1) According to the JKR formalism, we calculate the elastic energy of the system UE as the
difference between the elastic energies (UE)1 on loading and (UE)2 on unloading branches. Therefore, the stored elastic energy UE is UE = (UE)1 −(UE)2,
(4.2) (4.2) where (UE)1 = P1δ1 −
P1
0
δ(P) dP
(4.3) (4.3) at the loading of a curved axisymmetric punch until the fictitious load P1 that corresponds to the
true contact radius a1; and at the loading of a curved axisymmetric punch until the fictitious load P1 that corresponds to the
true contact radius a1; and (UE)2 =
P1
P0
δ(P) dP =
P1
P0
P
K(a1) dP = P2
1 −P2
0
2K(a1)
(4.4) (4.4) for a flat-ended punch of radius a1 that is unloaded from P1 to the true contact load P0. The above
solution (3.26) for a flat-ended circular punch has been used to calculate (4.4). Thus for a flat-ended punch of radius a1 that is unloaded from P1 to the true contact load P0. The above
solution (3.26) for a flat-ended circular punch has been used to calculate (4.4). Thus UE = (UE)1 −(UE)2 = P1δ1 −
P1
0
δ(P) dP −P2
1 −P2
0
2K(a1) . The mechanical work of the applied load is calculated as The mechanical work of the applied load is calculated as UM = −P0δ2 = −P0(δ1 − δ),
(4.5) (4.5) where δ = δ1 −δ2 is the change in the depth of penetration due to unloading. Taking into
account the solution (3.26), one obtains where δ = δ1 −δ2 is the change in the depth of penetration due to unloading. Taking into
account the solution (3.26), one obtains account the solution (3.26), one obtains δ = P1 −P0
K(a1) , δ = P1 −P0
K(a1) , and therefore, one has and therefore, one has UM = −P0δ2 = −P0
δ1 −P1 −P0
K(a1)
. (b) General expressions for power-law punches 12 12
rspa.royalsocietypublishing.org Proc. R.Soc. A 472: 20160550
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Let us consider the problem for a power-law punches whose shape is described by (3.13). It
follows from (3.15) that P = 2πmTBmam. (4.11) (4.11) Substituting (4.11) and (3.16) at a = a1 into (4.10), one get the expressions that solve the adhesive
contact problem for an arbitrary power-law punch of degree m Substituting (4.11) and (3.16) at a = a1 into (4.10), one get the expressions that solve the adhesive
contact problem for an arbitrary power-law punch of degree m P0 = 2πT
mBmam−1
1
−
2w
T
a1
(4.12) (4.12) and and δ2 = mBmam
1
ln
R
a1
+ 1
m
−
2w
T a1 ln
R
a1
. (4.13) (4.13) Evidently, the expressions (4.12) and (4.13) reduce to the above non-adhesive contact problems
(the analogy to the Hertz-type contact problems) in the case w = 0. on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from (4.9) (4.9) Further one has
P
P
√
8 T
K(
) δ Further one has
P1 −P0 = π
√
8wTa1 = K(a1) δ P1 −P0 = π
√
8wTa1 = K(a1) δ and hence, the following expression is valid and hence, the following expression is valid δ =
2w
T a1 ln
R
a1
. Thus, the general relations of the adhesive problem of contact between a graphene membrane
and an arbitrary convex, smooth blunt axisymmetric punch f(r), f(0) = 0, in the framework of the
JKR approach are P0 = P1 −π
√
8wTa1
and
δ2 = δ1 −
2w
T a1 ln
R
a1
. (4.10) (4.10) on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from (i) Real shapes of indenter tips Usually, for the experimental tests either special nanoindenters are used as the probe, or AFMs
having an indentation testing function [7]. An important point in such studies is the geometric
deviation of the probe shape from its nominal geometry, therefore, researchers employing
nanoindenters developed various empirical area functions to relate the apparent contact area to
depth. Borodich et al. [53] argued that at shallow depth, the nanoindenter blunt shapes are often
well described by power-law functions of degree m with 1 < m ≤2. This point was later discussed
by many researchers (for some references, see [44]). The same statement is applicable to actual
shapes of probes used for contact mode AFM. p
p
To describe the actual shapes of AFM tips, four different non-axisymmetric tips coated by
diamond-like carbon were studied using both a special tip characterizer and the 2D images
of the tip profiles obtained by scanning electron microscope. The values of m extracted from
the power law approximations of the AFM tip geometry z = −f(r, θ) ∼−Bm(r, θ)rm (z < 30 nm)
gave the values of m not close to 1 but rather m ≈2. In addition, one has to realize that (i)
nanoindenters drive the indenting tip always perpendicular to the surface during the depth-
sensing nanoindentation test, while loading of a probe through an AFM cantilever involves the
lateral movement and (ii) the AFM cantilever is mounted within an AFM device with a specific
tilt angle (see a discussion in [54]). Hence, even if the AFM tip may be nominally rather sharp,
the actual shape of a non-vertically fixed AFM tip near the first point of contact is rather blunt. Hence, the above expressions (4.12) and (4.13) can be used to describe the real adhesive contact
between a probe tip and a membrane of 2D material. (ii) Dimensionless expressions An elastic half-space itself does not have any characteristic scale. Hence, there are two
dilation similarity transformations of Hertz-type contact for a half-space [44,55]: (i) the punch
shape function is transformed by the same homogeneous dilations along all axes and (ii) the
punch shape function is transformed by dilation along the vertical axis only. The existence
of these similarity transformations is the reason that the 3D Hertz-type contact problems are
self-similar [56,57]. Owing to the absence of a characteristic scale, the choice of the characteristic parameters of
a 3D adhesive contact problem is rather arbitrary [27]. For example, as the characteristic scale
for a 3D adhesive contact problem, Borodich et al. [47] took the radius a1 of the contact region
at P0 = 0. For 2D membranes, the dimensionless variables could be also specified using various on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from 0
–0.4
–0.2
0
0.2
0.4
0.6
0.8
1.0
1.5
1.0
m = 2.00
m = 1.75
m = 1.25
m = 1.50
0.5
a1/a*
P0/P*
Figure 3. The dimensionless P0/P∗–a1/a∗relations (4.18) for monomial indenters of various degrees m. (Online version
in colour.) 0
–0.4
–0.2
0
0.2
0.4
0.6
0.8
1.0
1.5
1.0
m = 2.00
m = 1.75
m = 1.25
m = 1.50
0.5
a1/a*
P0/P* 13 Figure 3. The dimensionless P0/P∗–a1/a∗relations (4.18) for monomial indenters of various degrees m. (Online version
in colour.) characteristic scales, in particular, via the radius of the membrane. It follows from (4.12) that in
the adhesive contact problem for a 2D material, the non-zero radius a1 of the contact region at
P0 = 0 is characteristic scales, in particular, via the radius of the membrane. It follows from (4.12) that in
the adhesive contact problem for a 2D material, the non-zero radius a1 of the contact region at
P0 = 0 is a1(0) =
2w
Tm2B2m
1/2(m−1)
,
m > 1. (4.14) (4.14) One can see that a1(0) does not depend on the radius of the drum R. If this value is taken as
a characteristic size of the contact region in order to write dimensionless parameters, then the
characteristic parameters of the adhesive contact problems may be taken as a∗= a1(0),
P∗=
π2(m−1)23m−2wmTm−2
m2B2m
1/2(m−1)
and
δ∗=
2mwm
m2TmB2m
1/2(m−1)
. (ii) Dimensionless expressions (4.15) (4.15) In this case (4.12) and (4.13) have the following expressions P0
P∗=
a1
a∗
m
−
a1
a∗
(4.16) (4.16) and and δ2
δ∗=
a1
a∗
m 1
m −ln
a1/a∗
R/a∗
+ a1
a∗ln
a1/a∗
R/a∗
. (4.17) (4.17) If we denote ¯P = P0/P∗, ¯a = a1/a∗and ¯δ = δ2/δ∗, then (4.16) and (4.17) can be written as the
following dimensionless relations ¯P = ¯am −¯a
(4.18) ¯P = ¯am −¯a (4.18) and ¯δ = ¯am
1
m −ln
¯a
¯R
+ ¯a ln
¯a
¯R
,
¯R = R
a∗
(4.19) (4.19) that are valid for an arbitrary axisymmetric monomial punch of degree m > 1. The above case of power-law shaped punches is important for probing membranes by
indenters. Because the P0–a1 relation does not depend on the radii of the drum R, then in turn,
the ¯P–¯a one does not also depend on R. The graphs of the dimensionless ¯P–¯a relations (4.21) for
power-law indenters whose degree m are within the 0.25 ≤m ≤2 range are shown in figure 3. on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from 1
rspa.royalsocietypublishing.org Proc. R.Soc. A 472: 20160550
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . –2
0
2
0
0.5
1.0
1.5
2.0
2.5
18
10
12
14
16
4
6
8
d/d *
a/a*
m = 2.00
m = 1.75
m = 1.25
m = 1.50
Figure 4. The dimensionless ¯a–¯δ relations for power-law indenters for m within the 0.25 ≤m ≤2 range. (Online version
in colour.) –2
0
2
0
0.5
1.0
1.5
2.0
2.5
18
10
12
14
16
4
6
8
d/d *
a/a*
m = 2.00
m = 1.75
m = 1.25
m = 1.50 14 Figure 4. The dimensionless ¯a–¯δ relations for power-law indenters for m within the 0.25 ≤m ≤2 range. (Online version
in colour.) –2
–1
0
1
2
3
4
–0.4
–0.2
0
0.2
0.4
0.6
0.8
1.0
5
d/d *
P/P*
m = 2.00
m = 1.75
m = 1.25
m = 1.50
Figure 5. (d) Spherical punches The particular case of a spherical punch is very important for various applications. Let the punch
be of radius Rs, then f(r) = B2r2, m = 2 and B2 = 1/(2Rs). For non-adhesive contact, one has For non-adhesive contact, one has p(r) =
2T/Rs = const. for r ≤a
0
for r > a. (4.22) (4.22) It follows from (3.9) and (4.22) (or from (3.15)) that
πa2 2T
Rs
= P
or
a =
PRs
2πT
(4.23) It follows from (3.9) and (4.22) (or from (3.15)) that .22) (or from (3.15)) that
πa2 2T
Rs
= P
or
a =
PRs
2πT
(4.23) (4.23) and one gets from (3.16) the following nonlinear expression for δ(a) under a sphere of radius Rs and one gets from (3.16) the following nonlinear expression for δ(a) under a sphere of rad δ = a2
2Rs
2 ln
R
a
+ 1
. (4.24) (4.24) For adhesive contact, one can use the above general solution for monomial punches. It follows
from (4.15): a∗= Rs(2w/T)1/2, P∗= 4πRsw and δ∗= 2Rsw/T. Therefore, one obtains from (4.20)
and (4.21): (¯a)c = 1
2
and
( ¯P0)c = −1
4. Thus, for a spherical punch, one gets explicit expressions for the values of the critical contact
radius, displacement and the corresponding pull-off force in dimensional form ac = Rs
w
2T ,
δc = Rsw
4T
1 + 2 ln
Rs
R
w
2T
,
and
Pc = −πRsw. on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from Thus, the curve P–δ
describes the depth-sensing indentation of the system under
consideration. It is clear that if the external compressive load is not reduced then the indenter
and the membrane jump out of the contact at the point dP/dδ = 0, i.e. at the point of the tangent
line is horizontal in experiments at fixed load P (e.g. [23,44]). This point can be calculated from
the following relations 15 rspa.royalsocietypublishing.org Proc. R.Soc. A 472: 20160550
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . dP
dδ = dP/da
dδ/da = 0
or
dP
da = 0. It follows from (4.18) that for m > 1 the punch separates from the membrane at the critical
radius (¯a)c (¯a)c = m1/(1−m)
(4.20) (4.20) and at the corresponding critical load ¯Pc and at the corresponding critical load ¯Pc (¯P)c = (¯a)m
c −(¯a)c = mm/(1−m) −m1/(1−m) = m1/(1−m)(m−1 −1). (4.21) (4.21) (ii) Dimensionless expressions The dimensionless ¯P–¯δ relations for power-law indenters for m within the 0.25 ≤m ≤2 range. (Online version
in colour.) –2
–1
0
1
2
3
4
–0.4
–0.2
0
0.2
0.4
0.6
0.8
1.0
5
d/d *
P/P*
m = 2.00
m = 1.75
m = 1.25
m = 1.50 Figure 5. The dimensionless ¯P–¯δ relations for power-law indenters for m within the 0.25 ≤m ≤2 range. (Online version
in colour.) The graphs of the dimensionless ¯a–¯δ relation for power-law indenters whose degree m are
within the 0.25 ≤m ≤2 range are shown in figure 4. Using the above ¯P–¯a and ¯a–¯δ relations, one
can get the graphs of the dimensionless ¯P–¯δ relation for power-law indenters whose degree m
is within the 0.25 ≤m ≤2 range (figure 5). These and the above graphs have been obtained for
R/a = ¯R/¯a = 100. One can see that in the case of blunt smooth punches, the general character of
the new relations for 2D materials is similar to the character of the corresponding relations for
elastic 3D materials [44,47,48]. 5. Conclusion It has been argued that to study mechanical properties of atomically thin materials such as
graphene or graphene oxide, their discrete structure may be modelled as a stretched elastic
membrane whose bending rigidity is neglected. For such membranes, we have formulated
and solved axisymmetric contact problems that are analogous to the Hertz and Boussinesq
frictionless contact problems. For these new problems, a general statement for the slope of the
force–displacement curve has been formulated and proved. The results may be used not only in
application to monoatomic thick 2D materials but also to membranes of other types, e.g. few-layer
graphene sheets [2], the graphene oxide paper [58] and other thin films. Because attractive surface forces may be very significant at nanometer scale, analogies to the
JKR and the Boussinesq–Kendall contact problems in the presence of adhesive interactions have
been formulated. General nonlinear equations are derived for the relations between the actual
external force, the probe displacements and the contact radius in the case of an arbitrary smooth,
convex axisymmetric indenter. Note that the pressure distribution under action of a rigid punch
has the delta-function singularity on the circular line of the contact radius a1. Note that we have studied the problems applicable to the real tips of both nanoindenter
and AFM probes that may be normally described by power law functions of degrees m > 1. We have excluded from our consideration the adhesive contact problem for a cone (m = 1). This
case will be studied in another publication. For power-law shaped axisymmetric indenters, the
dimensionless form of the equations has been analysed, and the explicit expressions are derived
for the values of the pull-off force and for the corresponding critical contact radius. The particular
cases of spherical indenters and flat-ended circular indenters have been considered in detail. The
approach presented may be a ground for development new techniques for experimental studies
of mechanical properties of 2D materials. p
p
As it has been noted by an anonymous reviewer, considering that all of the problems discussed
have radial symmetry, equations (3.5) and (3.6) could have been developed without reference to
any Green’s function. Equally, the jump condition across r = a could have been given directly,
and all of the solutions (non-adhesive and adhesive) could have been derived directly from
the ordinary differential equation and the boundary and jump conditions. on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from From the equilibrium equation (2.3), one has From the equilibrium equation (2.3), one has 16 rspa.royalsocietypublishing.org Proc. R.Soc. A 472: 20160550
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . dUT
da1
= 0 = −2wπa1 +
P2
c
4πTa1
(4.27) (4.27) and, hence, one may obtain the adherence force (the pull-off force) of a flat-ended circular punch
of radius a1 Pc = 2πa1
√
2wT. (4.28) (4.28) Studying the frictionless Boussinesq–Kendall adhesive contact problem for an elastic-half
space, Maugis [23] came to the conclusion that the adherence force is proportional neither to the
energy of adhesion nor to the area of the contact. One can see from (4.28) that the same conclusion
is valid for a circular elastic membrane, but the adherence force is proportional to the perimeter
of the punch. (e) An analogy to the Boussinesq–Kendall adhesive contact problem Consider an axisymmetric flat-ended punch of radius a1 that is vertically pressed into an elastic
membrane. The elastic material deforms according to (3.26), i.e. δ = P0/K. The surface energy
is given as above by (2.2). Using (3.26), one obtains that the stored elastic energy UE and the
mechanical energy of the applied load UM are, respectively, UE =
P0
0
P dδ = P2
0
2K
and
UM = −P0δ = −P2
0
K . (4.25) (4.25) The total energy UT can be obtained by summation of all components UT = −wπa2
1 −P2
2K. (4.26) (4.26) on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from References 1. Meyer JC, Geim AK, Katsnelson MI, Novoselov KS, Booth TJ, Roth S. 2007 The structure of
suspended graphene sheets. Nature 446, 60–63. (doi:10.1038/nature05545) 1. Meyer JC, Geim AK, Katsnelson MI, Novoselov KS, Booth TJ, Roth S. 2007 The structure of
suspended graphene sheets. Nature 446, 60–63. (doi:10.1038/nature05545) 2. Novoselov KS, Geim AK, Morozov SV, Jiang D, Zhang Y, Dubonos SV, Grigorieva IV,
Firsov AA. 2004 Electric field effect in atomically thin carbon films. Science 306, 666–669. (doi:10.1126/science.1102896) 2. Novoselov KS, Geim AK, Morozov SV, Jiang D, Zhang Y, Dubonos SV, Grigorieva IV,
Firsov AA. 2004 Electric field effect in atomically thin carbon films. Science 306, 666–669. (doi:10.1126/science.1102896) 3. Stankovich S, Dikin DA, Dommett GHB, Kohlhaas KM, Zimney EJ, Stach EA, Piner RD,
Nguyen ST, Ruoff RS. 2006 Graphene-based composite materials. Nature 442, 282–286. (doi:10.1038/nature04969) 3. Stankovich S, Dikin DA, Dommett GHB, Kohlhaas KM, Zimney EJ, Stach EA, Piner RD,
Nguyen ST, Ruoff RS. 2006 Graphene-based composite materials. Nature 442, 282–286. (doi:10.1038/nature04969) (
)
4. Suk JW, Piner RD, An J, Ruoff RS. 2010 Mechanical properties of monolayer graphene oxide. ACSNano 4, 6557–6564. (doi:10.1021/nn101781v@proofing) 4. Suk JW, Piner RD, An J, Ruoff RS. 2010 Mechanical properties of monolayer graphene oxide. ACSNano 4, 6557–6564. (doi:10.1021/nn101781v@proofing) p
g
5. Castro Neto A, Novoselov K. 2011 Two-dimensional crystals: Beyond graphene. Mater. Express
1, 10–17. (doi:10.1166/mex.2011.1002) 5. Castro Neto A, Novoselov K. 2011 Two-dimensional crystals: Beyond graphene. Mater. Express
1, 10–17. (doi:10.1166/mex.2011.1002) 6. Castro Neto A, Novoselov K. 2011 New directions in science and technology: two-
dimensional crystals. Rep. Prog. Phys. 74, 082501. (doi:10.1088/0034-4885/74/8/082501) y
p
g
y
7. Castellanos-Gomez A, Singh V, van der Zant HSJ, Steele GA. 2015 Mechanics of freely-
suspended ultrathin layered materials. Ann. Phys. 527, 27–44. (doi:10.1002/andp.201400153) 8. Bolotin KI, Sikes KJ, Hone J, Stormer HL, Kim P. 2008 Temperature-dependent transport
in suspended graphene. Phys. Rev. Lett. 101, 096802–096804. (doi:10.1103/PhysRevLett. 101.096802) )
9. Geim AK. 2009 Graphene: status and prospects. Science 324, 1530–1534. (doi:10.1126/
science.1158877) 10. Chen C, Rosenblatt S, Bolotin KI, Kalb W, Kim P, Kymissis I, Stormer HL, Heinz TF, Hone J. 2009 Performance of monolayer graphene nanomechanical resonators with electrical readout. Nat. Nanotechnol. 4, 861–867. (doi:10.1038/nnano.2009.267) 11. Zhang Y, Zhao YP. 2014 Detecting the mass and position of an adsorbate on a drum resonator. Proc. R. Soc. A 470, 20140418. (doi:10.1098/rspa.2014.0418) p
12. Aguilera-Servin J, Miao T, Bockrath M. on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from the results on experimental studies of actual shapes of tips of industrial AFM probes. We thank also two
anonymous reviewers for their valuable comments. the results on experimental studies of actual shapes of tips of industrial AFM probes. We thank also two
anonymous reviewers for their valuable comments. the results on experimental studies of actual shapes of tips of industrial AFM probes. We thank also two
anonymous reviewers for their valuable comments. 17 17
rspa.royalsocietypublishing.org Proc. R.Soc. A 472: 20160550
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5. Conclusion We agree with this
comment, however, we would like to add that we intend to use the Green’s function approach in
our future papers for solving non-axisymmetric contact problems. Authors’ contributions. F.M.B. and B.A.G. contributed equally. Competing interests. The authors declare no competing interests. Funding. Supported by the Leverhulme Trust. Acknowledgements. The work was initiated as a part of activities of the CARBTRIB International Network
supported by the Leverhulme Trust. The authors are grateful to the Leverhulme Trust for the support of their
collaboration. Thanks are due to Dr E. Brousseau and Mrs Z.N.R. Alraziqi (Cardiff University) for sharing pp
y
Acknowledgements. The work was initiated as a part of activities of the CARBTRIB International Network
supported by the Leverhulme Trust. The authors are grateful to the Leverhulme Trust for the support of their
collaboration. Thanks are due to Dr E. Brousseau and Mrs Z.N.R. Alraziqi (Cardiff University) for sharing on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from 25. Johnson KL, Kendall K, Roberts AD. 1971 Surface energy and the contact of elastic solids. Proc. R. Soc. Lond. A 324, 301–313. (doi:10.1098/rspa.1971.0141) 18 p
26. Derjaguin BV, Muller VM, Toporov YP. 1975 Effect of contact deformations on adhesion of
particles. J. Colloid Interface Sci. 53, 314–326. (doi:10.1016/0021-9797(75)90018-1) 27. Borodich FM, Galanov BA. 2008 Non-direct estimations of adhesive and elastic properties
of materials by depth-sensing indentation. Proc. R. Soc. A 464, 2759–2776. (doi:10.1098/
rspa.2008.0044) p
)
28. Borodich FM, Galanov BA, Gorb SN, Prostov MY, Prostov YI, Suarez-Alvarez MM. 2013
Evaluation of adhesive and elastic properties of polymers by the BG method. Macromol. React. Eng. 7, 555–563. (doi:10.1002/mren.201300107) g
29. Shanahan MER. 2000 Adhesion of a punch to a thin membrane. C. R. Acad. Sci. Paris Ser. IV 1,
517–522. (doi:10.1016/S1296-2147(00)00147-5) (
(
)
)
30. Shull KR. 2002 Contact mechanics and the adhesion of soft solids. Mater. Sci. Eng. R Rep. 36,
1–45. (doi:10.1016/S0927-796X(01)00039-0) (
(
)
)
31. Berinskii IE, Borodich FM. 2013 On the isotropic elastic properties of graphene crystal lattice. Surf. Effects Solid Mech. 30, 33–42. (doi:10.1007/978-3-642-35783-1_3) f
ff
32. Derjaguin BV, Fedoseev DV. 1975 The synthesis of diamond at low pressure. Sci. Am. 233,
102–109. (doi:10.1038/scientificamerican1175-102) f
ff
32. Derjaguin BV, Fedoseev DV. 1975 The synthesis o
102–109. (doi:10.1038/scientificamerican1175-102) 33. Radushkevich LV, Lukyanovich VM. 1950 Structure of sorbents owing by the evidence
provided by an electron microscopy study. Zh. Fiz. Khim. 24, 21–42. (in Russian). p
y
py
y
34. Radushkevich LV, Lukyanovich VM. 1952 Structure of the carbon produced in the thermal
decomposition of carbon monoxide on an iron catalyst. Zh. Fiz. Khim. 26, 88–95. (in Russian). 35. Derjaguin BV, Fedoseev DV. 1977 Growth of diamond and graphite from the gas phase. Moscow,
R
i
N
k
(i
R
i
) 34. Radushkevich LV, Lukyanovich VM. 1952 Structure of the carbon produced in the thermal
decomposition of carbon monoxide on an iron catalyst. Zh. Fiz. Khim. 26, 88–95. (in Russian). decomposition of carbon monoxide on an iron catalyst. Zh. Fiz. Khim. 26, 88–95. (in Russian). 35. Derjaguin BV, Fedoseev DV. 1977 Growth of diamond and graphite from the gas phase. Moscow,
Russia: Nauka (in Russian). p
y
35. Derjaguin BV, Fedoseev DV. 1977 Growth of diamond and graphite from the gas phase. Moscow,
Russia: Nauka (in Russian). 36. Geim AK, Novoselov KS. 2007 The rise of graphene. Nat. Mat. 6, 183–191. References 2015 Nanoscale pressure sensors realized from
suspended graphene membrane devices. Appl. Phys. Lett. 106, 083103. (doi:10.1063/1.4908176) p
g
p
pp
y
13. Frank IW, Tanenbaum DM, van der Zande AM, McEuen PL. 2007 Mechanical properties of
suspended graphene sheets. J. Vaccum Sci. Technol. B 25, 2558–2561. (doi:10.1116/1.2789446) p
g
p
14. Lee C, Wei X, Kysar JW, Hone J. 2008 Measurement of the elastic properties and intrinsic
strength of monolayer graphene. Science 321, 385–388. (doi:10.1126/science.1157996) 15. Castellanos-Gomez A, Poot M, Steele GA, van der Zant HSJ, Agraït N, Rubio-Bollinger
G. 2012 Elastic properties of freely suspended MoS2 nanosheets. Adv. Mater. 24, 772–775. (doi:10.1002/adma.201103965) (
)
16. Timoshenko S, Woinowsky-Krieger S. 1959 Theory of plates and shells. New York, NY: McGraw
Hill. 17. Hooke R. 1667 Micrographia: or some physiological descriptions of minute bodies made by magnifying
glasses with observations and inquiries thereupon. London, UK: John Martin and James Allestry. 17. Hooke R. 1667 Micrographia: or some physiological descriptions of minute bodies made by magnifying
glasses with observations and inquiries thereupon. London, UK: John Martin and James Allestry. 18. Lebedew P. 1894 Ueber die mechanische Wirkung der Wellen auf ruhende Resonatoren. I. Electromagnetischen Wellen. Ann. Phys. 288, 621–640. (doi:10.1002/andp.18942880803) 18. Lebedew P. 1894 Ueber die mechanische Wirkung der Wellen auf ruhende Resonatoren. I. Electromagnetischen Wellen. Ann. Phys. 288, 621–640. (doi:10.1002/andp.18942880803) 19. Deryagin BV, Krotova NA, Smilga VP. 1978 Adhesion of solids. New York, NY: Consultants
Bureau. 20. Kendall K. 2001 Molecular adhesion and its applications. New York, NY: Kluwer Academic/
Plenum Publishers. 21. Ambrosetti A, Ferri N, DiStasio RA, Tkatchenko A. 2016 Wavelike charge density
fluctuations and van der Waals interactions at the nanoscale. Science 351, 1171–1176. (doi:10.1126/science.aae0509) 22. Derjaguin B. 1934 Untersuchungen über die Reibung und Adhäsion, IV. Theo
Anhaftens kleiner Teilchen. Kolloid Z. 69, 155–164. (doi:10.1007/BF01433225) (
)
23. Maugis D. 2000 Contact, adhesion and rupture of elastic solids. Berlin, Germany: Springe 24. Goryacheva IG, Makhovskaya YuYu. 2008 Adhesion effects in contact interaction of solids. Comptes Rendus Mécanique 336, 118–125. (doi:10.1016/j.crme.2007.11.003) on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from (doi:10.1038/
nmat1849) 37. Bunch JS, van der Zande AM, Verbridge SS, Frank IW, Tanenbaum DM, Parpia JM, Craighead
HG, McEuen PL. 2007 Electromechanical resonators from graphene sheets. Science 315,
490–493. (doi:10.1126/science.1136836) (
)
38. Gomez-Navarro C, Burghard M, Kern K. 2008 Elastic properties of chemically derived single
graphene sheets. Nano Lett. 8, 2045–2049. (doi:10.1021/nl801384y) 39. Krivtsov AM. 2009 Theoretical mechanics. Elastic properties of single-atomic and two-atomic crystals. Saint Petersburg, Russia: Polytechnical University Publisher (in Russian). 40. Berinskii IE, Krivtsov AM. 2010 On using many-particle interatomic potentials to compute
elastic properties of graphene and diamond. Mech. Solids 45, 815–834. (doi:10.3103/
S0025654410060063) )
41. Berinskii IE, Borodich FM. 2013 Elastic in-plane properties of 2D linearized models of
graphene. Mech. Mater. 62, 60–68. (doi:10.1016/j.mechmat.2013.03.004) g
p
j
42. Gillis P. 1984 Calculating the elastic constants of graphite. Carbon 22, 387–391. (doi:10.1016/
0008-6223(84)90010-1) 43. Hertz H. 1882
Ueber die Berührung fester elastischer Körper. J. Reine Angew. Math. 92,
156–171. [English transl. Hertz H. (1896) On the contact of elastic solids. In Miscellaneous Papers
by H. Hertz (eds DE Jones, GA Schott), pp. 146–162. London, UK: Macmillan. y
pp
44. Borodich FM. 2014 The Hertz-type and adhesive contact problems for depth
indentation. Adv. Appl. Mech. 47, 225–366. (doi:10.1016/B978-0-12-800130-1.00003-5) 45. Boussinesq J. 1885 Applications des potentiels à l’ etude de l’ equilibre et du mouvement des solides
elastique. Paris, France: Gauthier-Villars. q
,
46. Sperling G. 1964 Eine Theorie der Haftung von Feststoffteilchen an festen Körpern. Dissertation, Technische Hochschule Karlsruhe, Germany. y
47. Borodich FM, Galanov BA, Suarez-Alvarez MM. 2014 The JKR-type adhesive contact
problems for power-law shaped axisymmetric punches. J. Mech. Phys. Solids 68, 14–32. (doi:10.1016/j.jmps.2014.03.003) j j
p
48. Borodich FM, Galanov BA, Keer LM, Suarez-Alvarez MM. 2014 The JKR-type adhesive
contact problems for transversely isotropic elastic solids. Mech. Mater. 75, 34–44. (doi:10.1016/
j.mechmat.2014.03.011) j
49. Karman TV, Biot MA. 1940 Mathematical methods in engineering. An introduction to the
mathematical treatment of engineering problems. New York, NY: McGraw Hill. 50. LeDret H, Lucquin B. 2016 Partial differential equations: modeling, analysis and numerical
approximation. Basel, Switzerland: Birkhauser. on November 29, 2016
http://rspa.royalsocietypublishing.org/
Downloaded from 51. Polyanin AD. 2002 Handbook of linear partial differential equations for engineers and scientists. Boca
Raton, FL: Chapman and Hall/CRC. 19 p
. Dwight HR. 1961 Tables of integrals and other mathematical data. New York, NY: Macmillan rspa.royalsocietypublishing.org Proc. R.Soc. A 472: 20160550
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53. Borodich FM, Keer LM, Korach CS. 2003 Analytical study of fundamental nanoindentation
test relations for indenters of non-ideal shapes. Nanotechnology 14, 803–808. (doi:10.1088/
0957-4484/14/7/319) 54. Al-Musawi RSJ, Brousseau EB, Geng Y, Borodich FM. 2016 Insight into mechanics of AFM
tip-based nanomachining: Bending of cantilevers and machined grooves. Nanotechnology 27,
385302. (doi:10.1088/0957-4484/27/38/385302) 54. Al-Musawi RSJ, Brousseau EB, Geng Y, Borodich FM. 2016 Insight into mechanics of AFM
tip-based nanomachining: Bending of cantilevers and machined grooves. Nanotechnology 27,
385302. (doi:10.1088/0957-4484/27/38/385302) (
)
55. Borodich FM. 1989 Hertz contact problems for an anisotropic physically nonlinear elastic
medium. Strength Mater. 21, 1668–1676. (doi:10.1007/BF01533408) 55. Borodich FM. 1989 Hertz contact problems for an anisotropic physically nonlinear elastic
medium. Strength Mater. 21, 1668–1676. (doi:10.1007/BF01533408) 56. Galanov BA. 1981 Approximate solution to some problems of elastic contact of two bodies. Mech. Solids 16, 61–67. 56. Galanov BA. 1981 Approximate solution to some problems of elastic contact of two bodies. Mech. Solids 16, 61–67. 57. Borodich FM. 1983 Similarity in the problem of contact between elastic bodies. PMM J. Appl. Math. Mech. 47, 519–521. (doi:10.1016/0021-8928(83)90077-1) 57. Borodich FM. 1983 Similarity in the problem of contact between elastic bodies. PMM J. Appl. Math. Mech. 47, 519–521. (doi:10.1016/0021-8928(83)90077-1) 58. Dikin DA, Stankovich S, Zimney EJ, Piner RD, Dommett GHB, Evmenenko G, Nguyen ST,
Ruoff RS. 2007 Preparation and characterization of graphene oxide paper. Nature 448, 457–460. (doi:10.1038/nature06016) 58. Dikin DA, Stankovich S, Zimney EJ, Piner RD, Dommett GHB, Evmenenko G, Nguyen ST,
Ruoff RS. 2007 Preparation and characterization of graphene oxide paper. Nature 448, 457–460. (doi:10.1038/nature06016)
|
https://openalex.org/W4205974076
|
https://mediatum.ub.tum.de/doc/1651513/document.pdf
|
English
| null |
Effects of Cage Position and Light Transmission on Home Cage Activity and Circadian Entrainment in Mice
|
Frontiers in neuroscience
| 2,022
|
cc-by
| 2,240
|
EDITORIAL
published: 13 January 2022
doi: 10.3389/fnut.2021.835535 EDITORIAL published: 13 January 2022
doi: 10.3389/fnut.2021.835535 INTRODUCTION The benefits of regular physical activity and a healthy diet are well-documented in the literature
(1, 2). However, efforts to counter the global obesity epidemic and related metabolic diseases
through interventions focusing on either physical activity or diet alone have been of limited success
(3, 4), highlighting the need for new, more refined and integrated approaches. Although it appears
intuitive that interventions combining both lifestyle components have the capacity to result in
greater health benefits than singular approaches (5), it is also possible that changing behavior related
to one component may result in compensatory changes in the other. For example, the obvious
health benefits of increased physical activity may be overridden when the resulting increase in
energy expenditure is compensated by an increase in dietary energy intake and unhealthier food
choices (6). Likewise, dietary interventions such as caloric restriction may result in compensatory
reductions in physical activity behavior (7), which could negatively affect physical fitness and
performance, undermine weight loss success, and expose individuals to greater risk for future
weight regain and other associated diseases (8–10). Editorial on the Research Topic Understanding the Interaction Between Physical Activity and Diet for the Promotion of Health
and Fitness Karsten Koehler 1* and Clemens Drenowatz 2 Karsten Koehler 1* and Clemens Drenowatz 2 1 Department of Sport and Health Sciences, Technical University of Munich, Munich, Germany, 2 Division of Sport, Physical
Activity and Health, University of Education Upper Austria, Linz, Austria Keywords: exercise, eating behavior, weight loss, lifestyle, long-term health, energy balance Edited and reviewed by:
David Christopher Nieman,
Appalachian State University,
United States *Correspondence:
Karsten Koehler
karsten.koehler@tum.de *Correspondence:
Karsten Koehler
karsten.koehler@tum.de The goal of this Research Topic was to strengthen our understanding of how physical activity
and diet are related with each other in the context of health and fitness promotion. As such, this
Research Topic includes research targeting physical activity, which refers to any bodily movement
that results in energy expenditure, as well as exercise, a subset of physical activity with the goal
of maintaining or improving physical fitness (11). The Research Topic combines a total of 9
original studies and systematic reviews, which cover three basic themes ranging from the interplay
between nutrition and physical activity for weight loss (theme 1), the impact of exercise on food
intake regulation (theme 2) and the potential for negative health consequences of excessive exercise
(theme 3). Specialty section:
This article was submitted to
Sport and Exercise Nutrition,
a section of the journal
Frontiers in Nutrition
Received: 14 December 2021
Accepted: 20 December 2021
Published: 13 January 2022 Specialty section:
This article was submitted to
Sport and Exercise Nutrition,
a section of the journal
Frontiers in Nutrition Specialty section:
This article was submitted to
Sport and Exercise Nutrition,
a section of the journal
Frontiers in Nutrition Received: 14 December 2021
Accepted: 20 December 2021
Published: 13 January 2022 THEME 1: WEIGHT LOSS AND BODY COMPOSITION Koehler K and Drenowatz C (2022)
Editorial: Understanding the
Interaction Between Physical Activity
and Diet for the Promotion of Health
and Fitness. Front. Nutr. 8:835535. doi: 10.3389/fnut.2021.835535 Despite considerable efforts, global rates of overweight, and obesity continue to rise and excess body
weight is considered a major threat to future public health (12). Accordingly, various strategies,
including attempts to alter diet and physical activity, have been implemented to tackle the obesity January 2022 | Volume 8 | Article 835535 Frontiers in Nutrition | www.frontiersin.org Editorial: Physical Activity and Diet Interaction Koehler and Drenowatz epidemic. In their systematic review Correia et al. highlight
beneficial effects of intermittent fasting on body weight. Diet-induced changes in body weight, however, are generally
short lived and even greater benefits can be accomplished
with the inclusion of exercise training. Exercise also plays
an important role for maintaining lean body mass during
caloric restriction as indicated by Roth et al. who reported
a decline in lean mass during caloric restriction despite
assuring a high protein intake. In addition to structured
programs, lifestyle adjustments also play a critical role in
weight loss. Myers et al. show that particularly vigorous
physical activity, along with a reduction in energy-dense
foods, was associated with a more pronounced weight loss
in women. Similarly, van Baak et al. report greater weight
loss during caloric restriction in participants who increase
their physical activity. Furthermore, increased physical activity
was associated with beneficial changes in various cardio-
metabolic risk factors and weight loss maintenance beyond the
intervention period. thereby providing an avenue for reducing disease burden later
in life. THEME 3: POSSIBLE NEGATIVE
CONSEQUENCE Besides the beneficial effects of physical activity and exercise
there
are
also
some
possible
harmful
effects
that
need
to be acknowledged. Ribeiro et al. discuss the potential
detrimental effects of exacerbated exercise on the gastrointestinal
environment that can, among others, impair gastric motility, and
nutrient absorption. Moore et al. further address the increased
risk for low energy availability in endurance athletes, and
emphasize the need for adequate dietary energy intake during
periods of high energy demands. The overall benefits of physical
activity, however, should not be questioned by these results. Rather, these studies highlight the complex interaction between
diet and physical activity and their effects on the human body
and health. SUMMARY This overview of current research related to physical activity and
diet highlights the importance of integrating both components
regardless whether the goal is to maximize weight loss or to
diminish the potential negative effects at the upper end of the
activity spectrum. A deeper understanding of the interaction
between these two critical lifestyle approaches is required for
the development of combined interventions involving physical
activity and diet that result in successful, long-term health
improvements while avoiding unhealthy compensatory behavior
in the other domain. While physical activity and exercise increase energy expenditure,
most exercisers increase their dietary energy intake. This
phenomenon, often referred to compensatory eating, was
also described in a recent study by Horner et al. who
showed that neither gastric emptying nor appetite-regulating
hormones were significantly altered after a 4-week exercise
intervention, suggesting that short-term changes in gastro-
intestinal regulation play no major role in compensatory
eating. Post-exercise food intake was also studied by Okada
et al. who reported that administration of exogenous ketones
impacted appetite-regulating hormones but failed to affect
appetite perception and post-exercise energy intake. These two
interventions are complemented by a systematic review and
meta-analysis by Hubner et al. on the effects of exercise
on
appetite
regulation
in
older
adults. Despite
limited
research in this demographic, exercise, and physical activity
appear to promote satiety sensitivity and appetite control, AUTHOR CONTRIBUTIONS KK and CD wrote the introduction and the summary. CD
summarized the publications pertaining to weight loss and
excessive exercising. KK wrote the summaries of publications
relating to the impact of exercise on food intake regulation. Both
authors approved the submitted version. 8. Weiss EP, Racette SB, Villareal DT, Fontana L, Steger-May K, Schechtman
KB, et al. Lower extremity muscle size and strength and aerobic capacity
decrease with caloric restriction but not with exercise-induced weight loss. J
Appl Physiol. (2007) 102:634–40. doi: 10.1152/japplphysiol.00853.2006 9. Wang X, Lyles MF, You T, Berry MJ, Rejeski WJ, Nicklas BJ. Weight regain
is related to decreases in physical activity during weight loss. Med Sci Sports
Exerc. (2008) 40:1781–8. doi: 10.1249/MSS.0b013e31817d8176 old) who are overfat, or obese; systematic review and meta-analysis. J Diabetes
Metab Disord. (2015) 14:31. doi: 10.1186/s40200-015-0154-1
6. King NA, Horner K, Hills AP, Byrne NM, Wood RE, Bryant E, et al. Exercise,
appetite and weight management: understanding the compensatory responses
in eating behaviour and how they contribute to variability in exercise-induced
weight loss. Br J Sports Med. (2012) 46:315–22. doi: 10.1136/bjsm.2010.082495
7. Redman LM, Heilbronn LK, Martin CK, de Jonge L, Williamson DA, Delany
JP, et al. Metabolic and behavioral compensations in response to caloric
restriction: implications for the maintenance of weight loss. PLoS One. (2009)
4:e4377. doi: 10.1371/journal.pone.0004377
8. Weiss EP, Racette SB, Villareal DT, Fontana L, Steger-May K, Schechtman
KB, et al. Lower extremity muscle size and strength and aerobic capacity
decrease with caloric restriction but not with exercise-induced weight loss. J
Appl Physiol. (2007) 102:634–40. doi: 10.1152/japplphysiol.00853.2006
9. Wang X, Lyles MF, You T, Berry MJ, Rejeski WJ, Nicklas BJ. Weight regain
is related to decreases in physical activity during weight loss. Med Sci Sports
Exerc. (2008) 40:1781–8. doi: 10.1249/MSS.0b013e31817d8176 old) who are overfat, or obese; systematic review and meta-analysis. J Diabetes
Metab Disord. (2015) 14:31. doi: 10.1186/s40200-015-0154-1 7. Redman LM, Heilbronn LK, Martin CK, de Jonge L, Williamson DA, Delany
JP, et al. Metabolic and behavioral compensations in response to caloric
restriction: implications for the maintenance of weight loss. PLoS One. (2009)
4:e4377. doi: 10.1371/journal.pone.0004377 6. King NA, Horner K, Hills AP, Byrne NM, Wood RE, Bryant E, et al. Exercise,
appetite and weight management: understanding the compensatory responses
in eating behaviour and how they contribute to variability in exercise-induced
weight loss. Br J Sports Med. (2012) 46:315–22. doi: 10.1136/bjsm.2010.082495 REFERENCES old) who are overfat, or obese; systematic review and meta-analysis. J Diabetes
Metab Disord. (2015) 14:31. doi: 10.1186/s40200-015-0154-1 1. Warburton
DER,
Bredin
SSD. Health
benefits
of
physical
activity:
a
systematic
review
of
current
systematic
reviews. Curr
Opin
Cardiol. (2017)
32:541–56. doi:
10.1097/HCO.000000000000
0437 1. Warburton
DER,
Bredin
SSD. Health
benefits
of
physical
activity:
a
systematic
review
of
current
systematic
reviews. Curr
Opin
Cardiol. (2017)
32:541–56. doi:
10.1097/HCO.000000000000
0437 6. King NA, Horner K, Hills AP, Byrne NM, Wood RE, Bryant E, et al. Exercise,
appetite and weight management: understanding the compensatory responses
in eating behaviour and how they contribute to variability in exercise-induced
weight loss. Br J Sports Med. (2012) 46:315–22. doi: 10.1136/bjsm.2010.082495 6. King NA, Horner K, Hills AP, Byrne NM, Wood RE, Bryant E, et al. Exercise,
appetite and weight management: understanding the compensatory responses
in eating behaviour and how they contribute to variability in exercise-induced
weight loss. Br J Sports Med. (2012) 46:315–22. doi: 10.1136/bjsm.2010.082495 2. Yu E, Malik VS, Hu FB. Cardiovascular disease prevention by diet
modification: JACC health promotion series. J Am Coll Cardiol. (2018)
72:914–26. doi: 10.1016/j.jacc.2018.02.085 2. Yu E, Malik VS, Hu FB. Cardiovascular disease prevention by diet
modification: JACC health promotion series. J Am Coll Cardiol. (2018)
72:914–26. doi: 10.1016/j.jacc.2018.02.085 3. Freire R. Scientific evidence of diets for weight loss: different macronutrient
composition, intermittent fasting, and popular diets. Nutrition. (2020)
69:110549. doi: 10.1016/j.nut.2019.07.001 4. Swift DL, McGee JE, Earnest CP, Carlisle E, Nygard M, Johannsen NM. The
effects of exercise and physical activity on weight loss and maintenance. Prog
Cardiovasc Dis. (2018) 61:206–13. doi: 10.1016/j.pcad.2018.07.014 5. Clark JE. Diet, exercise or diet with exercise: comparing the effectiveness of
treatment options for weight-loss and changes in fitness for adults (18-65 years January 2022 | Volume 8 | Article 835535 Frontiers in Nutrition | www.frontiersin.org 2 Editorial: Physical Activity and Diet Interaction Koehler and Drenowatz Publisher’s Note: All claims expressed in this article are solely those of the authors
and do not necessarily represent those of their affiliated organizations, or those of
the publisher, the editors and the reviewers. Any product that may be evaluated in
this article, or claim that may be made by its manufacturer, is not guaranteed or
endorsed by the publisher. 10. Locher JL, Goldsby TU, Goss AM, Kilgore ML, Gower B, Ard JD. Calorie
restriction in overweight older adults: do benefits exceed potential risks? Exp
Gerontol. (2016) 86:4–13. doi: 10.1016/j.exger.2016.03.009 Frontiers in Nutrition | www.frontiersin.org 11. Caspersen CJ, Powell KE, Christenson GM. Physical activity, exercise, and
physical fitness: definitions and distinctions for health-related research. Public
Health Rep. (1985) 100:126–31. January 2022 | Volume 8 | Article 835535 10. Locher JL, Goldsby TU, Goss AM, Kilgore ML, Gower B, Ard JD. Calorie
restriction in overweight older adults: do benefits exceed potential risks? Exp
Gerontol. (2016) 86:4–13. doi: 10.1016/j.exger.2016.03.009
11. Caspersen CJ, Powell KE, Christenson GM. Physical activity, exercise, and
physical fitness: definitions and distinctions for health-related research. Public
Health Rep. (1985) 100:126–31.
12. Hruby
A,
Hu
FB.
The
epidemiology
of
obesity:
a
big
picture.
Pharmacoeconomics. (2015) 33:673–89. doi: 10.1007/s40273-014-0243-x REFERENCES Publisher’s Note: All claims expressed in this article are solely those of the authors
and do not necessarily represent those of their affiliated organizations, or those of
the publisher, the editors and the reviewers. Any product that may be evaluated in
this article, or claim that may be made by its manufacturer, is not guaranteed or
endorsed by the publisher. j
g
11. Caspersen CJ, Powell KE, Christenson GM. Physical activity, exercise, and
physical fitness: definitions and distinctions for health-related research. Public
Health Rep. (1985) 100:126–31. 12. Hruby
A,
Hu
FB. The
epidemiology
of
obesity:
a
big
picture. Pharmacoeconomics. (2015) 33:673–89. doi: 10.1007/s40273-014-0243-x 12. Hruby
A,
Hu
FB. The
epidemiology
of
obesity:
a
big
picture. Pharmacoeconomics. (2015) 33:673–89. doi: 10.1007/s40273-014-0243-x Copyright © 2022 Koehler and Drenowatz. This is an open-access article distributed
under the terms of the Creative Commons Attribution License (CC BY). The use,
distribution or reproduction in other forums is permitted, provided the original
author(s) and the copyright owner(s) are credited and that the original publication
in this journal is cited, in accordance with accepted academic practice. No use,
distribution or reproduction is permitted which does not comply with these terms. Conflict of Interest: The authors declare that the research was conducted in the
absence of any commercial or financial relationships that could be construed as a
potential conflict of interest. January 2022 | Volume 8 | Article 835535 Frontiers in Nutrition | www.frontiersin.org 3
|
https://openalex.org/W2163099364
|
http://wrap.warwick.ac.uk/4427/1/WRAP_Danon_journal.pone.0006895.pdf
|
English
| null |
Use of Cumulative Incidence of Novel Influenza A/H1N1 in Foreign Travelers to Estimate Lower Bounds on Cumulative Incidence in Mexico
|
PloS one
| 2,009
|
cc-by
| 5,973
|
University of Warwick institutional repository: http://go.warwick.ac.uk/wrap This paper is made available online in accordance with
publisher policies. Please scroll down to view the document
itself. Please refer to the repository record for this item and our
policy information available from the repository home page for
further information. To see the final version of this paper please visit the publisher’s website. Access to the published version may require a subscription. Author(s): Marc Lipsitch, Martin Lajous, Justin J O'Hagan, Ted Cohen,
Joel C Miller, Edward Goldstein, Leon Danon, Jacco Wallinga, Steven
Riley, Scott F Dowell, Carrie Reed, Meg McCarron Article Title: Use of Cumulative Incidence of Novel Influenza A/H1N1
in Foreign Travelers to Estimate Lower Bounds on Cumulative Article Title: Use of Cumulative Incidence of Novel Influenza A/H1N1
in Foreign Travelers to Estimate Lower Bounds on Cumulative
Incidence in Mexico
Year of publication: 2009
Link to published article: Year of publication: 2009 University of Warwick institutional repository: http://go.warwick.ac.uk/wrap Abstract Background: An accurate estimate of the total number of cases and severity of illness of an emerging infectious disease is
required both to define the burden of the epidemic and to determine the severity of disease. When a novel pathogen first
appears, affected individuals with severe symptoms are more likely to be diagnosed. Accordingly, the total number of cases
will be underestimated and disease severity overestimated. This problem is manifest in the current epidemic of novel
influenza A/H1N1. Methods and Results: We used a simple approach to leverage measures of incident influenza A/H1N1 among a relatively
small and well observed group of US, UK, Spanish and Canadian travelers who had visited Mexico to estimate the incidence
among a much larger and less well surveyed population of Mexican residents. We estimate that a minimum of 113,000 to
375,000 cases of novel influenza A/H1N1 have occurred in Mexicans during the month of April, 2009. Such an estimate
serves as a lower bound because it does not account for underreporting of cases in travelers or for nonrandom mixing
between Mexican residents and visitors, which together could increase the estimates by more than an order of magnitude. Conclusions: We find that the number of cases in Mexican residents may exceed the number of confirmed cases by two to
three orders of magnitude. While the extent of disease spread is greater than previously appreciated, our estimate suggests
that severe disease is uncommon since the total number of cases is likely to be much larger than those of confirmed cases. Citation: Lipsitch M, Lajous M, O’Hagan JJ, Cohen T, Miller JC, et al. (2009) Use of Cumulative Incidence of Novel Influenza A/H1N1 in Foreign Travelers to
Estimate Lower Bounds on Cumulative Incidence in Mexico. PLoS ONE 4(9): e6895. doi:10.1371/journal.pone.0006895 ditor: Alison P. Galvani, Yale University, United States of America Received May 21, 2009; Accepted July 27, 2009; Published September 9, 2009 eived May 21, 2009; Accepted July 27, 2009; Published Septem sitch et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits
tion, and reproduction in any medium, provided the original author and source are credited. Copyright: 2009 Lipsitch et al. This is an open-access article distributed under the terms of the Creative Commons Attribut
unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract Funding: This work was supported by the US National Institutes of Health cooperative agreement 5U01GM076497 ‘‘Models of Infectious Disease Agent Study’’
(M.Lipsitch, J.O’H. and E.G.), the Mexican National Council for Science and Technology (CONACyT), the Ministry of Health of Mexico (M.Lajous) and the Department
of Epidemiology, Harvard School of Public Health (M.Lajous and J.O’H.), the RAPIDD program of the Science & Technology Directorate, Department of Homeland
Security, and the Fogarty International Center, National Institutes of Health (J.C.M.), the Medical Research Council, UK (L.D.) and the Research Fund for The Control
of Infectious Disease of the Government of the Special Administrative Region of Hong Kong, and NIH R01 TW008246-01 from the Ecology of Infectious Disease
program (SR). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. mpeting Interests: ML has received compensation for consulting for the Avian/Pandemic Flu Registry, funded in part by Roche. Competing Interests: ML has received compensation for consulting for the Avian/Pandemic Flu Registry, fu mpeting Interests: ML has received compensation for consulting for the Avian/Pandemic Flu Registry, funded in part by Roche. -mail: mlipsitc@hsph.harvard.edu * E-mail: mlipsitc@hsph.harvard.edu Use of Cumulative Incidence of Novel Influenza A/H1N1
in Foreign Travelers to Estimate Lower Bounds on
Cumulative Incidence in Mexico Marc Lipsitch1,3*, Martin Lajous2,3, Justin J. O’Hagan3, Ted Cohen3,4, Joel C. Miller3,5, Edward Goldstein3,
Leon Danon3,6, Jacco Wallinga7, Steven Riley8, Scott F. Dowell9, Carrie Reed10, Meg McCarron10 1 Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, Massachusetts, United States of America, 2 Center for Population Health
Research, National Institute of Public Health, Cuernavaca, Mexico, 3 Department of Epidemiology, Harvard School of Public Health, Boston, Massachusetts, United States of
America, 4 Division of Global Health Equity, Brigham and Women’s Hospital, Harvard School of Public Health, Boston, Massachusetts, United States of America, 5 Fogarty
International Center, National Institutes of Health, Bethesda, Maryland, United States of America, 6 Department of Biological Sciences, University of Warwick, Coventry,
United Kingdom, 7 Epidemiology and Surveillance Unit, Centre for Infectious Disease Control, National Institute of Public Health and the Environment (RIVM), Bilthoven,
The Netherlands, 8 School of Public Health and Department of Community Medicine, University of Hong Kong, Hong Kong, People’s Republic of China, 9 Division of
Global Disease Detection and Emergency Response, Centers for Disease Control and Prevention, Atlanta, Georgia, United States of America, 10 Influenza Division, National
Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, United States of America http://dx.doi.org/10.1371/journal.pone.0006895 Publisher statement: Lipsitch M, Lajous M, O'Hagan JJ,
Cohen T, Miller JC, et al. (2009) Use of Cumulative Incidence
of Novel Influenza A/H1N1 in Foreign Travelers to Estimate
Lower Bounds on Cumulative Incidence in Mexico. PLoS ONE
4(9): e6895. doi:10.1371/journal.pone.0006895 PLoS ONE | www.plosone.org Citation: Lipsitch M, Lajous M, O’Hagan JJ, Cohen T, Miller JC, et al. (2009) Use of Cumulative Incidence of Novel Influenza A/H
Estimate Lower Bounds on Cumulative Incidence in Mexico. PLoS ONE 4(9): e6895. doi:10.1371/journal.pone.0006895 September 2009 | Volume 4 | Issue 9 | e6895 Introduction By assuming
(conservatively) that the risk of infection experienced by Mexicans
is at least equal to that experienced by visitors, and using travel
data to assess the amount of person-time at risk for visitors, we
estimate the incidence rate in proportion to the Mexican
population, and estimate a lower-bound of how many cases may
have been present in Mexico at a defined time. Introduction lative incidence is often difficult to measure in a large epidemic,
because often there is a bias toward ascertainment of severe cases. lative incidence is often difficult to measure in a large epidemic,
because often there is a bias toward ascertainment of severe cases. Where
underreporting
of
asymptomatic
and
mild
cases,
especially those that do not present for medical care, is likely,
there is a need for nonstandard approaches to estimate the
magnitude of the epidemic and severity of disease. Here we
propose and apply such a method to estimate the number of cases
of novel influenza A/H1N1 in Mexico up to approximately April
30, 2009, based on the number of cases observed in foreign
travelers. Intuitively, the notion is that such travelers act as
‘‘canaries in the mine’’ who briefly experience the daily risk of Where
underreporting
of
asymptomatic
and
mild
cases,
especially those that do not present for medical care, is likely,
there is a need for nonstandard approaches to estimate the
magnitude of the epidemic and severity of disease. Here we
propose and apply such a method to estimate the number of cases
of novel influenza A/H1N1 in Mexico up to approximately April
30, 2009, based on the number of cases observed in foreign
travelers. Intuitively, the notion is that such travelers act as
‘‘canaries in the mine’’ who briefly experience the daily risk of A reliable estimate of the cumulative number of infections for an
emerging disease, such as novel influenza A/H1N1, is critical to
determine both the magnitude of the problem and the severity of
disease. Cumulative incidence is the most direct estimate of the
magnitude
of
the
epidemic,
while
cumulative
deaths
and
hospitalizations must be divided by cumulative incidence (with
appropriate correction for reporting delays and censoring [1]) to
estimate the probability of severe outcomes for individuals that
become infected. While critical for situational awareness, cumu- PLoS ONE | www.plosone.org September 2009 | Volume 4 | Issue 9 | e6895 September 2009 | Volume 4 | Issue 9 | e6895 1 H1N1 Incidence in Mexico be somewhat less than assumed here. We therefore do not include
this large estimate in our overall range of estimates. infection prevalent in Mexico during their visit, then return home
to areas where, given the elevated level of concern, they may be
detected as cases of novel H1N1, even if not severe. Baseline estimate We estimate that approximately 375,000 Mexicans were
infected with novel H1N1 influenza with symptom onset up to
approximately April 30, 2009. This estimate derives from 283
cases among US, UK, Spanish and Canadian travelers, counting
confirmed and probable cases for the US and confirmed cases only
for the other two countries. Citizens of these countries together
accounted for approximately 689,250 airplane passenger visits to
Mexico in the period April 1–30, 2009, and international visitors
to Mexico had a mean length of stay of approximately 3.5 days, for
a total of 2.4 million person-days of exposure during this period
(Table 1). This implies that visitors experienced an incidence rate
of 91 cases per million person-days at risk. In the same period, the
Mexican population of approximately 107 million persons had
306107 million, or 3.2 billion person-days of exposure. Sensitivity analysis: possible clusters among travelers,
and border state cases Several cases among travelers may have resulted from clusters of
exposure and/or from transmission within the traveling group. In
order to exclude the effects of transmission among travelers or
cases of disease imported by means other than air travel, we
provide a revised estimate calculated from a subset of 228 cases. This reduced number of cases excludes both secondary cases
within putative clusters of travelers (these data were available for
travelers from each country except Spain) and excludes US cases
residing in or south of the closest major city to the Mexican border
who may have visited by means other than air travel. This
approach yields an estimate of 302,000 cases in Mexicans;
additional correction for clustering in Spanish cases, if the
required data were available, would further reduce this figure. Here we estimate that at least 113,000–375,000 cases of novel
H1N1 influenza occurred in Mexicans before the end of April,
2009. We discuss the uncertainties associated with this estimate
and present our rationale for why this number represents a lower
bound for the true number. Finally, we discuss the implications for
estimating the case-fatality proportion of this infection in Mexico. Sensitivity analysis: length of stay For reasons discussed below, we believe that 3.5 days is an
appropriate estimate for the mean duration of stay in Mexico for
all visitors, which heavily weights US visitors because the US is the
largest source of visitors. However, given that one study suggests a
considerably longer length of stay [2], and that non-US visitors
likely stay longer given the longer trip involved, we performed a
sensitivity analysis assuming that visitors from the US, Canada and
European countries have lengths of stay of 8.7, 10.5, and 13.9 days
respectively, using numbers from an unpublished 2008 update of
the 2001–5 survey (Gerardo Vazquez, Mexico Ministry of
Tourism, personal communication). Using the data with possible
clusters and near-border cases removed, produces a low estimate
of 113,000 cases in Mexican residents. September 2009 | Volume 4 | Issue 9 | e6895 Sensitivity analysis: non-homogeneous disease across
Mexico y
y
y
Travel history was known for 49% (929/1890) of US confirmed
cases and 48% (86/179) of Canadian confirmed cases, 97% (37/
38) of the UK cases and 100% (93/93) of Spanish cases. If the
proportion of cases with travel history to Mexico is assumed to be
the same for those with missing data in this field, the imputed
number of total cases with travel history would rise to 418, and the
implied number of cases in Mexicans would rise to 554,000. We
strongly suspect that travel history is more likely to be known in
those who did travel to Mexico than in those who did not, which
would suggest that the correction for missing travel history should This
analysis
assumes
that
incidence
during
April
was
homogeneous across 107 million Mexicans. If the rates of disease
among Mexicans in travel destinations was higher or lower than
elsewhere, this might substantially alter these estimates. The
national cumulative incidence of suspect cases as of May 9 was
17.32/100,000, which was 16x higher than that in Puebla, the
state with the lowest incidence, and 4x lower than that in Distrito
Federal, the capital, with the highest reported incidence. Quintana
Roo, the state containing Cancun, which is the most popular Table 1. Cases of novel influenza A/H1N1 among travelers to Mexico from three countries as of May 6, 2009 (Canada) or May 8,
2009 (US, UK, Spain) and associated estimates. Table 1. Cases of novel influenza A/H1N1 among travelers to Mexico from three countries as of May 6, 2009 (Canada) or May 8,
2009 (US, UK, Spain) and associated estimates. Table 1. Cases of novel influenza A/H1N1 among travelers to Mexico from three countries as of May 6, 2009 (Canada) or May 8,
2009 (US, UK, Spain) and associated estimates. Discussion We have estimated that there are likely to have been at least
113,000–375,000 cases of novel H1N1 influenza among Mexicans
with onset during the month of April, 2009. Taking into account what
we consider to be extreme sensitivity analyses, this estimate could
change by approximately 2-fold in either direction. This exceeds the
number of confirmed cases reported to WHO, 1204 as of May 8, 2009
(http://www.who.int/csr/don/GlobalSubnationalMaster_20090508_
1815.jpg), by a factor of approximately 100 or more. We have shown in Table 1 the estimates obtained using only
travelers from each country individually. Here, the US-based
estimates are the lowest, with greater estimates from those based
on Canadians and still greater estimates based on Europeans. In
part this may reflect a longer duration of trips for travelers from
more distant destinations, but even using the destination-specific
duration data does not remove this effect. As we note below, we
cannot rule out the possibility that some transmission occurred on
airplanes; such transmission might be more likely in travelers flying
longer distances. Differences in patterns of exposure within
Mexico, chance variation and other factors must account for the
remaining differences. It is unsurprising that we estimate a larger number than the
number of cases confirmed in Mexico, since ascertainment there
has been particularly focused on severe cases. Nevertheless, we
regard this estimate as likely a lower bound on the actual number
of cases in Mexico, for two principal reasons. First, the analytic
approach assumes that the incidence rate in Mexicans in Mexico is
equal to that in travelers. If indeed the infection has been
transmitting extensively within Mexico, one would expect that the
exposure of travelers to the virus would be somewhat less than that
of residents, due to nonrandom mixing between residents and
travelers; travelers should be less exposed to residents than other
residents are. Prior models of influenza transmission (set in the
United States) have assumed that 36–51% of influenza transmis-
sion takes place outside of home or school [3]. One might roughly
estimate that this is the proportion of transmission to which both
visitors and residents would be exposed, suggesting that incidence
in residents might be 2–3x as high as that in visitors; however, this
approach has obvious limitations given the uncertainty of those
estimates and the fact that they were made for a different country. This simple model has several principal limitations. Discussion First, we do
not incorporate exposure of travelers who arrive by ship or
overland, only by air. While we have excluded from the numerator
the one traveler case with a known cruise ship exposure, we may
have slightly overestimated the incidence in travelers by neglecting
such exposures. Second, our calculations make the assumption that
incidence is uniform geographically throughout Mexico and across
age group. All but one state in Mexico have now reported cases
(http://portal.salud.gob.mx/sites/salud/descargas/pdf/influenza/
situacion_actual070511.pdf), and all have at least suspect cases [3],
so it is likely reasonable to assume that persons throughout Mexico
were exposed to some extent. However, the exposure may not have
been uniform. This may be a further reason to consider our estimate
as a lower bound, since the detected cases are heavily concentrated
in the State of Mexico and the Distrito Federal, the destination of
,18% of visitors from these countries, while the most popular
airport of entry for visitors from the US, UK and Canada in April
2009 was Cancun, which accounted for 47.5%–74.5% of visitors for
each nationality but had relatively low reported incidence. As the
pandemic has evolved, it has become clear that different age groups
experience different risks of confirmed and probable infection with
the pandemic virus, with the highest rates of confirmed and
probable infection among persons under 25 years old (http://www. cdc.gov/h1n1flu/surveillanceqa.htm). Finally, we assume that
transmission to travelers occurred in Mexico, not on an aircraft. An influenza outbreak on an aircraft has been documented [9], and
if a cluster of such infections were included in our numbers, it would
result in an overestimate of incidence in Mexico. Notably, 36% of
travel-associated cases in Spain for whom data were available were
symptomatic during the inbound flight; given the incubation period
of influenza, these travelers, at least, could not plausibly have
become infected during the flight [10]. Second, while most cases ascertained in the traveler population
to date have been mild, one nonetheless expects that many mild
cases (as well as probable but unconfirmed cases) in travelers are
absent from our calculations. A survey in New York City, where
case ascertainment was aggressive surrounding the St. Francis
School outbreak, indicated that over 1000 persons associated with
the school experienced influenza-like illness, in a period where
only 74 confirmed or probable cases were ascertained. Sensitivity analysis: non-homogeneous disease across
Mexico US (confirmed+probable)
Canada (confirmed)
UK (confirmed)
Spain (confirmed)
Total
Cases with Mexico travel history
132
62
19
70
283
Cases with travel history known/total cases
928/1890
86/179
37/38
93/93
With only one case per possible cluster, and
near border cases removed
85
56
17
no data to assess
clusters; 70 assumed
228
Travel volume for April
526,861
119,473
22,013
20,903
668,347
Inferred incidence rate (/million person-days)
72
148
246
957
117
Inferred cases in Mexico
229,000
475,000
789,000
3,062,000
375,000
Inferred incidence rate (/million person-days)*
18
44
55
241
35
Inferred cases in Mexico*
59,000
142,000
178,000
771,000
113,000
doi:10.1371/journal.pone.0006895.t001 A/H1N1 among travelers to Mexico from three countries as of May 6, 2009 (Canada) or May 8,
d estimates nza A/H1N1 among travelers to Mexico from three countries as of May 6, 2009 (Canada) or May 8,
iated estimates. September 2009 | Volume 4 | Issue 9 | e6895 2 PLoS ONE | www.plosone.org 2 H1N1 Incidence in Mexico proportion as the deaths reflect cases from an earlier, smaller
phase of the epidemic [6]. Also, counting only laboratory
confirmed deaths is likely to result in a significant underestimation
of the true number of deaths, because of insensitivity depending on
the timing and adequacy of the specimen, the fact that many
severe pneumonia patients were not tested (approximately 1000–
2000 such cases typically occur in Mexico in April [7]), and the
fact that a majority of influenza deaths are attributed to circulatory
causes rather than identified as pneumonia or influenza [8]. Nonetheless, as the number of deaths accumulates, especially if
illness onset dates are available for fatal cases, our estimates may
provide an appropriate denominator for revised estimates of the
case-fatality proportion. The number of hospitalizations associated
with suspect cases was 6,754 as of May 9 [4], which combines with
our denominator to give a hospitalization proportion of about 2%,
closer to figures observed elsewhere. single destination for travelers from these countries, reported
incidence of 12.10/100,000. If these incidence numbers reflect
true incidence variation in the country (which is unlikely to be the
only source of variation), then total Mexican incidence should be
1.4 times higher than that estimated from Cancun travelers, or 4x
lower than that estimated for Mexico City travelers. Unfortunate-
ly, destination data are not available for the majority of travel-
associated cases in any of the four countries we considered. PLoS ONE | www.plosone.org September 2009 | Volume 4 | Issue 9 | e6895 Discussion For our primary
analysis, however, we used figures from the Ministry of Tourism
indicating a mean length of stay of 3.4 days (see Methods), while
an independent study conducted by the National Association of
Hotels and Motels finds a similar value of 3.6 days for the mean
length of hotel stay by foreign visitors, and a very recent survey
found that the majority of US leisure travelers interested in visiting
Mexico take vacations for 4 nights or less (personal communica-
tion). Our travel volumes are lower in part because we have used
citizenship rather than first destination outside Mexico (to better
reflect likely final destination) and have used data on number of
incoming passengers (corrected to estimate outgoing passengers)
rather than flight data, which may perhaps reflect capacities rather
than actual numbers. Altogether, these differences in data sources
could account for approximately a 3-fold variation in estimates,
apart from the variation due to different time periods considered. content&view = article&id = 125&Itemid = 193. Estimates of the
number of travelers returning from Mexico during the period
April 1–30 were obtained using data from Mexican immigration
records deposited in the Sistema Integral de Operacio´n Migratoria
(SIOM). This database contains information on the citizenship of
all travelers arriving into Mexican airports. Assuming that the
populations of inbound and outbound travelers from Mexico are
in near-steady state the number of inbound travelers should give a
reliable estimate of the number of outbound travelers. Records
were abstracted for the period April 1–30. Note that our method is
not strongly sensitive to the exact period considered, since
additional days would proportionately increase the person-time
for Mexican residents and approximately proportionately increase
the person-time for visitors. We did not decrement the person-time
to account for time no longer at risk once a Mexican resident was
infected. The number of Canadian, British and Spanish travelers arriving
into Mexico began to drop off on April 27th, likely in response to
the media coverage of the outbreak, while the number of US
travelers to Mexico began to decrease on April 26th. As it is
unlikely that the number of outbound travelers decreased over this
period we calculated the average number of travelers arriving into
Mexico for each day of the week using data for the first three
weeks of April. These estimates were used instead of the actual
daily numbers of travelers for the latter days of April. Data sources Cases in travelers. Cases ascertained in the US in travelers
were obtained from the US CDC line list dated May 8 at 0100
EDT, reflecting cases reported up to May 7. Possible clusters of
traveler cases were detected by manual scan of the line list for cases
with common county of report, closely related onset dates, and no
indication
that
they
lived
in
different
households. Cases
ascertained in Canada in travelers were obtained from a copy of
the Canadian line list dated May 6 residing at the US CDC. Possible clusters of traveler cases were noted on the line list itself. Cases ascertained in the UK in travelers were obtained from a
comprehensive scan of press reports cross-checked with UK
Health Protection Agency daily updates to ensure consistency of
numbers, and possible clusters were ascertained the same way. One case from the United States known to be in a woman visiting
Mexico on a cruise ship was excluded since cruise ship visitors
were not included in our travel estimates. The number of cases in
travelers was denoted U. Use of line lists from 6–8 days after our
period of interest was selected because for those entering the US
CDC line list, the mean delay from symptom onset was 7 days. Hence, the US data, which represented the majority of cases,
should be representative of cases with onset in the period up to
April 30. The number of cases from Spain was taken from the
recent report produced by the Surveillance Group in Spain [10]. p
)
Alternative estimates obtained from a 2008 Bank of Mexico
tourism survey (Gerardo Vazquez, Mexico Ministry of Tourism,
personal communication) an earlier version of which was used by
Fraser et al. [11] give longer durations of stay overall and indicate
heterogeneity by nationality in length of stay: 8.7 nights for US
citizens, 10.5 nights for Canadians and 13.9 nights for others. These estimates were used in a sensitivity analysis. We note that
with a typical incubation period of about 1–2 days for influenza A
[13], individuals infected early on in a stay of two weeks would
have been sick for a week or more before returning home, at which
point they might have stopped shedding detectable virus. Discussion The total
number of travelers into Mexico was denoted Pt. The mean
duration of stay was assumed to be D~3.5 days. This was based
on a mean stay of 3.6 days from survey data for hotel stays in April
2009 from the National Association of Hotels and Motels of
Mexico (personal communication) and on a mean stay of 3.4 days
from survey data posted by the Mexican Tourism Ministry
(http://www.sectur.gob.mx/wb/secturing/sect_8978_study_of__
tourist_pr). In addition, a survey of a representative sample of US
leisure travelers interested in visiting Mexico conducted in
February and March of 2009 found that 74% of all vacations
taken by this group were 4 nights or less (P. Yesawich, National
Leisure Travel Monitor, personal communication). Accurate estimation of the magnitude of an emerging epidemic
is essential for maintaining situational awareness and determining
a rational public health response. The simple approach applied
here indicates that the likely number of cases of H1N1 influenza
among Mexican residents during the month of April, 2009 was at
least two orders of magnitude larger than that detected. While
such calculations should not be interpreted as precise estimates of
cumulative incidence, they provide important perspective in
interpreting data from detected cases in situations where extensive
surveillance is unlikely to occur. Data sources Our
estimates are based on infections confirmed in the country to
which a traveler returned, and would therefore tend to miss many
such infections, suggesting that only a fraction of such a long stay
would be ‘‘at risk’’ for the event of infection detected upon return. Analysis If the incidence rate in Mexicans were x times that in visitors,
then the following equality should hold, relating the incidence rate
in each population: x U
DPt ~
M
30PM, where in the month of April
each Mexican had 30 days at risk, and each visitor had D days at
risk on average. Estimates for each quantity except for M, the
unknown number of incident cases in Mexican residents, were
provided from data, under the conservative assumption that x~1,
and the equation was solved for M. The major statistical
uncertainty in our estimates comes from the number of visitors
who were infected, which as a count with a value of 283 should Discussion If these
figures reflect the typical rate of under-reporting in the United
States, then the inferred figures from Mexico should increase by
.1000/74 = 14-fold. Likewise, any foreign residents who became
ill in Mexico (rather than in their home country) may have been
missed in our counts of travelers. In essence, the method used here
is a way to estimate cases in a population where they are likely
being undercounted, based on travelers to countries in which
undercounting, though present, is less severe. Since the inferred
number of cases in Mexican residents scales linearly with the
number observed in travelers, the number in Mexican residents is
likely to be considerably higher than we have estimated. Forty-eight deaths were observed up to May 9 among
laboratory-confirmed cases in Mexico [4]. While it might be
tempting to calculate a case-fatality proportion by dividing this
number by the estimated number of cases in Mexico, such a
calculation would likely be misleading, for several reasons. In a
growing epidemic, given a significant delay from illness onset to
death
[5],
one
expects
to
underestimate
the
case-fatality Our estimates of cases are larger, by about 10-fold, than those
reported by Fraser et al. [11]. Importantly, this reflects the fact
that we base ascertainment on numbers available on May 6–8,
while Fraser et al. base ascertainment on numbers available on September 2009 | Volume 4 | Issue 9 | e6895 PLoS ONE | www.plosone.org September 2009 | Volume 4 | Issue 9 | e6895 3 H1N1 Incidence in Mexico April 30. With rapid epidemic growth, the difference of one week
is likely to account for a difference of perhaps 2-8-fold. Also, Fraser
et al. use a longer mean length of stay (9 days) and a larger travel
volume. Estimates of the length of stay cited by Fraser et al. [11]
were close to 9 days in 2001-5 [2], and we have considered a
sensitivity analysis based on an updated version of that survey,
using numbers specific to origin of the travelers. Discussion For our primary
analysis, however, we used figures from the Ministry of Tourism
indicating a mean length of stay of 3.4 days (see Methods), while
an independent study conducted by the National Association of
Hotels and Motels finds a similar value of 3.6 days for the mean
length of hotel stay by foreign visitors, and a very recent survey
found that the majority of US leisure travelers interested in visiting
Mexico take vacations for 4 nights or less (personal communica-
tion). Our travel volumes are lower in part because we have used
citizenship rather than first destination outside Mexico (to better
reflect likely final destination) and have used data on number of
incoming passengers (corrected to estimate outgoing passengers)
rather than flight data, which may perhaps reflect capacities rather
than actual numbers. Altogether, these differences in data sources
could account for approximately a 3-fold variation in estimates,
apart from the variation due to different time periods considered. Accurate estimation of the magnitude of an emerging epidemic
is essential for maintaining situational awareness and determining
a rational public health response. The simple approach applied
here indicates that the likely number of cases of H1N1 influenza
among Mexican residents during the month of April, 2009 was at
least two orders of magnitude larger than that detected. While
such calculations should not be interpreted as precise estimates of
cumulative incidence, they provide important perspective in
interpreting data from detected cases in situations where extensive
surveillance is unlikely to occur. April 30. With rapid epidemic growth, the difference of one week
is likely to account for a difference of perhaps 2-8-fold. Also, Fraser
et al. use a longer mean length of stay (9 days) and a larger travel
volume. Estimates of the length of stay cited by Fraser et al. [11]
were close to 9 days in 2001-5 [2], and we have considered a
sensitivity analysis based on an updated version of that survey,
using numbers specific to origin of the travelers. PLoS ONE | www.plosone.org Person-time at risk The Mexican population was assumed to be PM = 106,682,518
persons as estimated by the National Council for Population
of Mexico http://www.conapo.gob.mx/index.php?option = com_ PLoS ONE | www.plosone.org September 2009 | Volume 4 | Issue 9 | e6895 September 2009 | Volume 4 | Issue 9 | e6895 4 H1N1 Incidence in Mexico Barrios from City Express Hotels, Alejandro Vazquez from Posadas Hotels
and Peter Yesawich from Ypartnership for providing length of stay data. have a coefficient of variation of 6%, negligible compared to the
uncertainties of underreporting and differences in exposure of the
visitor and resident populations. For this reason, statistical
uncertainty was not explicitly quantified in our estimates. Author Contributions Conceived and designed the experiments: M. Lipsitch JM SR SD. Performed the experiments: M. Lipsitch. Analyzed the data: M. Lipsitch
JJO TC JM EG LD JW CR MM. Wrote the paper: M. Lipsitch. Obtained
the data: M. Lajous JJO CR MM. Conceived and designed the experiments: M. Lipsitch JM SR SD. Performed the experiments: M. Lipsitch. Analyzed the data: M. Lipsitch
JJO TC JM EG LD JW CR MM. Wrote the paper: M. Lipsitch. Obtained
the data: M. Lajous JJO CR MM. Acknowledgments We thank Lyn Finelli, Martin Cetron, and David Shay for assistance in
initiating this project, and Neil Ferguson for helpful comments. We also
thank Gerardo Vazquez from the Ministry of Tourism, Mexico, Luis 13. Lessler J, Reich NG, Brookmeyer R, Perl TM, Nelson KE, Cummings DA
(2009) Incubation periods of acute respiratory viral infections: a systematic
review. Lancet Infect Dis 9: 291–300. 7. Kuri-Morales P, Galvan F, Cravioto P, Zarraga Rosas L, Tapia-Conyer R
(2006) Mortalidad en Me´xico por influenza y neumonı´a (1990–2005). Salud
Publica Mex 48: 379–384. PLoS ONE | www.plosone.org 12. Ypartnership/Yankelovich I (2008) National Leisure Travel Monitor. 6. Anderson RM, Fraser C, Ghani AC, Donnelly CA, Riley S, et al. (2004)
Epidemiology, transmission dynamics and control of SARS: the 2002-2003
epidemic. Philos Trans R Soc Lond B Biol Sci 359: 1091–1105. 5. Brundage JF, Shanks GD (2008) Deaths from bacterial pneumonia during 1918-
19 influenza pandemic. Emerg Infect Dis 14: 1193–1199. September 2009 | Volume 4 | Issue 9 | e6895 References 8. Thompson WW, Shay DK, Weintraub E, Brammer L, Cox N, et al. (2003)
Mortality associated with influenza and respiratory syncytial virus in the United
States. JAMA 289: 179–186. 1. Ghani AC, Donnelly CA, Cox DR, Griffin JT, Fraser C, et al. (2005) Methods
for estimating the case fatality ratio for a novel, emerging infectious disease. Am J Epidemiol 162: 479–486. J
9. Moser MR, Bender TR, Margolis HS, Noble GR, Kendal AP, et al. (1979) An
outbreak of influenza aboard a commercial airliner. Am J Epidemiol 110: 1–6. y
(
)
p
3. Halloran ME, Ferguson NM, Eubank S, Longini IM, Jr., Cummings DA, et al. (2008) Modeling targeted layered containment of an influenza pandemic in the
United States. Proc Natl Acad Sci U S A 105: 4639–4644. 10. Surveillance Group for New Influenza A(H1N1) Virus Investigation and Control
in Spain (2009) New influenza A(H1N1) virus infections in Spain, April–May
2009. Eurosurveillance 14: 1–4. 4. Direccion General Adjunta de Epidemiologia; Ministerio de Salud de Mexico
(2009) Brote de Influenza Humana A H1N1 Mexico; Boletin Diario No. 14, 09/
05/09. 11. Fraser C, Donnelly CA, Cauchemez S, Hanage WP, Van Kerkhove MD, et al. (2009) Pandemic potential of a strain of influenza A(H1N1): Early findings. Science 324: 1557–61. 5. Brundage JF, Shanks GD (2008) Deaths from bacterial pneumonia during 1918-
19 influenza pandemic. Emerg Infect Dis 14: 1193–1199. 6. Anderson RM, Fraser C, Ghani AC, Donnelly CA, Riley S, et al. (2004)
Epidemiology, transmission dynamics and control of SARS: the 2002-2003
epidemic. Philos Trans R Soc Lond B Biol Sci 359: 1091–1105. 13. Lessler J, Reich NG, Brookmeyer R, Perl TM, Nelson KE, Cummings DA
(2009) Incubation periods of acute respiratory viral infections: a systematic
review. Lancet Infect Dis 9: 291–300. PLoS ONE | www.plosone.org September 2009 | Volume 4 | Issue 9 | e6895 5
|
https://openalex.org/W4306721768
|
https://www.researchsquare.com/article/rs-1909456/latest.pdf
|
English
| null |
Coconut endocarp shell ash (CESA): a non-conventional catalyst for green synthesis of 2-amino-4H-benzochromenes
|
Research on chemical intermediates
| 2,022
|
cc-by
| 5,931
|
Coconut endocarp shell ash (CESA): a non-
conventional catalyst for green synthesis of 2-
amino-4H-benzochromenes Sandip P. Patil
PDVP College, (Affiliated to Shivaji University
Sachinkumar K. Shinde
PDVP College, (Affiliated to Shivaji University
Suresh S. Patil
(
sanyujapatil@yahoo.com
)
PDVP College, (Affiliated to Shivaji University Sandip P. Patil
PDVP College, (Affiliated to Shivaji University
Sachinkumar K. Shinde
PDVP College, (Affiliated to Shivaji University
Suresh S. Patil
(
sanyujapatil@yahoo.com
)
PDVP College, (Affiliated to Shivaji University Research Article Keywords: Coconut endocarp shell, Ash, Multicomponent reaction, Naphthols, Green chemistry
Posted Date: August 4th, 2022
DOI: https://doi.org/10.21203/rs.3.rs-1909456/v1
License:
This work is licensed under a Creative Commons Attribution 4.0 International
License. Read Full License Page 1/21 Page 1/21 Abstract In the present research, the coconut endocarp shell as a natural waste was converted into a valuable non-
conventional catalyst by simple thermal treatment, namely coconut endocarp shell ash (CESA). The
prepared CESA was characterized using several analytical techniques, including TGA, FT-IR, XRD, BET,
SEM, and EDX. The catalytic property of the CESA was examined in the synthesis of pharmaceutically
active 2-amino-4H-benzochromene derivatives via one-pot three component condensation of aldehydes,
malononitrile, and α or β-naphthol in ethanol at room temperature. The most promising aspects of the
presented approach are the high catalytic efficiency, short reaction time, excellent yield of products,
operational simplicity, waste to wealth, and optimization with the design of experiment. Moreover, the
CESA catalyst was recycled five times without any apparent loss of its catalytic activity. Introduction In the last few decades, green chemistry principles have played an important role in protecting the
environment. Thus, modern synthetic chemists make continuous efforts to come up with green principles
while conducting organic transformation with eco-friendly processes, especially eco-friendly catalyst [1–
3]. In general, green chemistry principles focus on the design and application of clean and safe
procedures in order to maximize efficiency and productivity, use renewable bio-degradable materials,
minimize by-products and hazardous reagents, which reduce the use of hazardous chemicals that are
risky to the environment and human beings. From this point of view, some aspects of green chemistry,
such as multicomponent reactions and reusable catalyst, can be combined, effectively following green
chemistry protocols can be expected [4, 5]. Multicomponent reaction (MCR) is an efficient, powerful technique that achieves a significant role in
modern synthetic organic chemistry due to the fact that all the required reactants are incorporated into
one pot to produce the desired products in a single step without isolation of any intermediate. Further,
using a one-pot, multi-component reaction approach could result in several advantages such as
operational simplicity, flexibility, shortage of synthetic routes, atom economy, decreased pollution, low
cost, straightforward reaction design, and simple purification of target products. Moreover, MCRs with a
heterocyclic moiety are particularly used in the construction of biologically active scaffolds and drug-like
molecules, which have increased interest in the research community [6–9]. In addition, 2-amino-4H-benzochromene represents an important class of heterocyclic structural motifs. It
is found in a number of natural products with remarkable biological active molecules [10], and varied
pharmacological properties such as anti-tumor or tumor vascular disrupting agent [11], anti-proliferative
[12, 13], cytotoxic [14], anti-cancer and SRC kinase inhibitory action [15], anti-bacterial [16], anti-fungal
[17], and anti-rheumatic actions [18]. Figure 1 represents some of the bioactive 2-amino-4H-
benzochromene compounds that possess different types of pharmaceutical activities. Introduction The present research work is to
explore that, naturally sourced catalytic system is a promising alternative tool in organic transformation. Herein, we report the first time a straightforward, cheap, and environmentally benign application of
coconut endocarp shell ash (CESA) as a recoverable green catalyst, for synthesizing 2-amino-4H-
benzochromene, from the condensation of β- or α-naphthol, aromatic aldehydes with malononitrile at
room temperature in ethanol solvent (Scheme 1). This is a clean, efficient synthetic method, that
preserves the simplicity as well as consistently gives good to excellent yields of corresponding products. In continuation of our present research, focusing on replacing inefficient traditional protocols with more
efficient, cost-effective, and environmentally friendly alternatives [50–54]. The present research work is to
explore that, naturally sourced catalytic system is a promising alternative tool in organic transformation. Herein, we report the first time a straightforward, cheap, and environmentally benign application of
coconut endocarp shell ash (CESA) as a recoverable green catalyst, for synthesizing 2-amino-4H-
benzochromene, from the condensation of β- or α-naphthol, aromatic aldehydes with malononitrile at
room temperature in ethanol solvent (Scheme 1). This is a clean, efficient synthetic method, that
preserves the simplicity as well as consistently gives good to excellent yields of corresponding products. Introduction Page 2/21 Page 2/21 A literature survey revealed that, because of the biological activity of these compounds, by one-pot three
component reaction of aryl aldehyde, malononitrile, and β- or α-naphthol, several improved protocols have
been investigated to synthesize 2-amino-4H-benzochromene by employing various catalysts, such as
piperidine [19], TBABr [20], basic alumina [21], hexadecyl trimethyl ammonium bromide (HTMAB) [22],
TiCl4 [23], NaOH [24], Amberlyst A-21 [25], KF-Al2O3[26], 1,4-diazabicyclo[2.2.2]octane [27], Mg/Al
hydrotalcite [28], Na2HPO4[29], Fe(HSO4)3[30], thiourea dioxide [31], p-dimethylaminopyridine [32], basic
ionic liquid [33], Nano zeolite clinoptilolite [34], PANDMAPF [35], Ni-Al2O3 [36], SILLP [37], potassium 2-
oxoimidazolidine-1,3-diide (POImD) [38], also some nanocatalyst or ionic liquid supported nanocatalyst
have been used for these transformations include CNT-Fe3O4 IL [39], SiO2@Im-Fc[OAc] [40], Fe3O4@PS-
Arg MNPs [41], γ- Fe2O3@HAp@CPTMS@AT [42], and GO Fc@Fe3O4 [43], However, for the synthesis of 2-
amino-4H-benzochromenes, many reported methods have effective and their own merits, but they suffer
from some drawbacks such as harsh reaction conditions, prolonged reaction times, use of toxic and
expensive solvents, difficult workup procedures, and furnishing the products in unsatisfactory yields. Also, the catalyst plays an important role in organic transformation for selectivity and determining yield. In recent years, the use of agro-food waste material as a catalyst has received significant attention in
green protocols for multi-component synthesis. Therefore, natural feedstock as catalyst offers a
promising alternative tool for organic processes due to their ability to act as catalysts with large surface
area, biodegradability, low toxicity, low cost, and reusability properties [44, 45]. Therefore, the main goal of the current research work is to investigate the organic transformation through
the utilization of naturally sourced catalysts. So, to avoid the problem of disposal of solid waste biomass
and to minimize possible environmental pollution, the naturally waste biomass used for the development
of promising catalysts to make synthetic protocols cheap can be considerably reduced [46]. Cocas
nucifera is a plant from the Areceae family (Palm family). On over 10 million hectares of land, coconuts
are grown all over the world. India, Indonesia, and the Philippines account for over 75% of global coconut
production [47, 48]. According to literature, the coconut shell is rich in organic substance, having 33.61%
cellulose, 36.51% lignin, 29.27% pentose, and 0.61% ash and inorganic matter [49]. In continuation of our present research, focusing on replacing inefficient traditional protocols with more
efficient, cost-effective, and environmentally friendly alternatives [50–54]. Experimental Procedure Materials And Methods Page 3/21 Page 3/21 All chemicals and laboratory-grade reagents used for this study were obtained from Sigma-Aldrich and
Merck chemical companies and were utilized without further purification. An analytical thin layer
chromatography (TLC) technique was used to check the purity of substances, and the progress of the
reaction was performed using Merck 0.2 mm silica gel 60 F-254 Al-plates. The confirmation and
characterization of the isolated pure form of products by spectroscopic data (IR, 1H NMR, 13C NMR, mass
spectra). Infrared (FT-IR) spectra were obtained in KBr pellets on a Bruker ALPHA FT-IR spectrometer. Brucker-AC spectrometers in CDCl3 and DMSO as solvent (for 1H NMR at 300 MHz and for 13C NMR at 75
MHz) are used to record 1H and 13C NMR spectral data. A Shimadzu QP2010 GCMS with an iron
temperature of 280°C was used to obtain the mass spectra. The melting points of purified derivatives
were determined using the DBK programmable melting point as an open capillary method. Thermal
gravimetric analysis (TGA) was measured using a TA SDT Q600 V20.9 Build instrument in the presence
of air at a linear heating rate of 10 oC min− 1 from 25 to 1000 oC. For measurement of the scanning
electron microscope (SEM), a JEOL (Tokyo, Japan) JSM-5200 was used. The N2 adsorption-desorption
isotherm for BET was recorded with a Micromeritics ASAP 2010 surface area and porosity analyser. An
energy-dispersive X-ray spectroscopy analysis without oxygen was recorded with the Quanta 200 3D FEI
scanning electron microscope. Preparation Of Coconut Endocarp Shell Ash (Cesa) Catalyst To prepare the CESA catalyst, bio-waste endocarp shells of Cocos nucifera were collected from the local
food market. The collected coconut endocarp shells (CES) (Fig. 2a) were washed by using distilled water
to remove dirt and sun dried for two days. The dried CES (50 g) were broken into small pieces by using a
mortar and pestle (Fig. 2b) and heated in a muffle furnace up to 800°C for two hours in a silica crucible. A
fine soft ash (Fig. 2c) was obtained and was named as coconut endocarp shell ash (CESA) (Fig. 2c). Further, CESA was used as a catalyst for the synthesis of 2-amino-4H-benzochromenes. DSC-TGA analysis The thermal behavior of coconut endocarp shell ash (CESA) was investigated by using thermo
gravimetric analysis (TGA) inflow of nitrogen atmosphere. The usual sample loading was 7-8 mg (Fig. 3). The TGA analysis of CESA shows the temperatures at which it is decomposed when heated in a
controlled environment. At temperatures that ranged from 0 to 1000 °C, four distinct stages of weight loss
were observed. The results reveal that, CESA showed the observed weight change below 100 oC. In the
first step, minor weight loss at temperatures below 130.70 °C was 2.915 %, and in the second step,
another weight loss occurred between 130.70 °C and 177.70 °C was 2.509 % due to the loss of adsorbed
moisture. In the third step, weight loss between temperature ranges of 177.70 °C and 627.76 °C was due
to the decomposition of chlorides and phosphates. The decomposition of metal carbonates caused the
10.90% weight loss in the fourth step between the temperatures of 627.76 °C and 740.23 °C, while metal
oxides and silica were responsible for the 71.34% weight of residue remaining after the temperatures of
984.10 °C. FTIR analysis Fourier transform infrared (FT-IR) analysis was used to determine the active functional groups in the
CESA catalyst. In the FT-IR spectrum of CESA (Fig. 4), the strong absorption bands occurred in the range
of 2382 cm-1, 2311 cm-1, 1515 cm-1, 1028 cm-1, 628 cm-1, clearly show that the presence of metal
carbonates and the absorption band at 1695 cm-1 indicates the presence of the C=O group of carbonate. Also, the weak absorption stretching band that appears at 3362 cm-1 can be attributed to the presence of
the OH group in the CESA catalyst that supports the formation of metal hydroxides due to the absorption
of moisture from the environment. XRD analysis The phase composition of the synthesized CESA catalyst was analysed by X-ray diffraction (Fig. 5). After
thermal treatment with synthesized CESA, it can be seen that the peaks appeared at 2θ = 28.08°, 28.84°,
29.86°, 30.19°, 44.87°, 47.17°, 49.89°, 68.54°, and 75.66° hint the presence of characteristic peaks of
metal carbonates. The peaks appearing at 2θ = 31.01°, 33.59°, 34.65°, 40.60°, 41.05°, and 43.12° were
characteristic peaks of metal oxides. Metal hydroxides were found at 2θ = 20.95°, 21.09°, and 22.01°. Thus, the conversion of reactant into product, these active phases of the catalyst were utilized. Characterization of coconut endocarp shell ash (CESA) catalyst To identify the active sites of the CESA catalyst, it was characterized by using different analytical
methods, including DSC-TGA, FT-IR, XRD, EDX, BET, and SEM. General procedure for the synthesis of 2-amino-4 H -benzochromene derivatives In a clean and dry 25 mL round-bottom flask, a CESA catalyst (10 wt. %) was added to the mixture of aryl
aldehyde 1 (1 mmol), malononitrile 2 (1 mmol), β-naphthol 3a or α-naphthol 3b (1 mmol) and ethanol. Then, at room temperature, the reaction mixture was magnetically stirred for an appropriate time until the
desired product was formed completely. The reaction progress was monitored by TLC (ethyl acetate:
hexane 1:9). After the reaction was completed, the reaction mixture was treated for solvent extraction
using ethyl acetate (4 x 10 mL). The combined organic phase was first washed with water, dried
(Na2SO4), and the solvent was removed under reduced pressure to obtain a crude product. The further
obtained crude products were purified by recrystallization using 96% ethanol. The identities of the
synthesized products were established by FT-IR, 1H NMR, and 13C NMR spectroscopic methods. Additionally, the melting points of the synthesized products were measured, and they agree with values
previously reported elsewhere in the literature. Additionally, the melting points of the synthesized products were measured, and they agree with values
previously reported elsewhere in the literature. Page 4/21 Page 4/21 Page 4/21 EDX analysis Page 5/21 Page 5/21 Page 5/21 The type of elements in coconut endocarp shell ash (CESA) was investigated by recording EDX. It is
shown in Fig. 6. The report reveals that this catalyst included the oxides of K, Na, Si, and Ca elements
present with uniform distributions and moderate to less distributions of elements, including Mg, Mn, Fe
and Cl. The carbonates, oxides, and hydroxides of corresponding metals gives basicity to the CESA
catalyst which plays a crucial role in the catalyst. The type of elements in coconut endocarp shell ash (CESA) was investigated by recording EDX. It is
shown in Fig. 6. The report reveals that this catalyst included the oxides of K, Na, Si, and Ca elements
present with uniform distributions and moderate to less distributions of elements, including Mg, Mn, Fe
and Cl. The carbonates, oxides, and hydroxides of corresponding metals gives basicity to the CESA
catalyst which plays a crucial role in the catalyst. BET analysis Fig. 7 indicates the data related to surface area, mean pore volume, and average pore radius for the
activated CESA catalyst. The surface area and mean pore volume of the CESA catalyst were 168.92 m2/g
and 0.093536 cm3/g-1, respectively. The mean pore radius was 2.215 nm, respectively. It was found to be
active, which provides a better catalytic surface area to catalyse the reaction effectively. SEM analysis The synthesized CESA catalyst was analyzed by employing scanning electron microscopy (SEM) to
determine the surface morphology and particle shape as represented in Fig. 8. These SEM images clearly
illustrate the CESA particles were observed with irregular sizes; they also have a porous nature due to their
smooth and soft active surface area, which provides a higher catalytic activity. 3.2 Optimization of the reaction conditions % of CESA catalyst in 5 mL of ethanol solvent is sufficient to show the reactivity of
the reactants (1 mmol each), and the corresponding product 4b was obtained at a 98% yield within 05
min. (Table 1, entry 9). Furthermore, the optimization of the model reaction was investigated by using
MeOH, THF, CH3CN, and H2O as solvents, and the yield of the desired product was obtained from 42-84%
(Table 1, entries 12-16). Thus, the best yield of the desired product was achieved in optimal ethanol
solvent (5 mL) by using 10 wt. % of CESA catalyst at room temperature (Table 1, entry 9). After finding the promising optimal reaction conditions for the model reaction, to extend the scope of
catalytic activity of the CESA catalyst, to prepare 2-amino-4H-benzochromene using the reaction between
various aryl aldehydes (1a-p), malononitrile (2) with β-naphthol (3a), were investigated under optimum
conditions. The results are presented in Table 2. According to the results in Table 2, the presence of
electron –withdrawing or releasing groups on their aryl aldehyde ring was successfully participated in
optimal reaction conditions and gave the products with excellent yields 4a-m. It was noticed that aryl
aldehydes having electron withdrawing groups had somewhat faster reaction rates than those with
electron-releasing groups. It was also discovered that the yields of meta- and para-substituted aryl
aldehydes were higher than those of ortho-substituted aryl aldehydes. Generally, this reaction is
straightforward, and the desired product precipitates out of the reaction mixture. Isolation of the catalyst
or evaporation of organic solvent from the reaction mixture affords pure products using simple filtration. All the desired products 4a-m of this reaction give good to excellent yields within short reaction times
under the optimized reaction conditions. The structural confirmation of the isolated pure product 4a-m
was investigated by melting point, IR, 1H NMR, and 13C NMR spectral data. The excellent performance of the synthesized CESA catalyst in the synthesis of biologically active 2-
amino-4H-benzochromenes (4a-m) from various aldehydes, malononitrile and β-naphthol encouraged us
to study its effect on the synthesis of 2-amino-4H-benzochromenes (5a-l) by replacement of the β-
naphthol with α-naphthol. Several derivatives of the 2-amino-4H-benzochromenes have been synthesized
and the results are presented in Table 2. It was discovered that the CESA catalyst provides a good to
excellent yield of the desired product in a shorter reaction time. 3.2 Optimization of the reaction conditions After this initial success, the efficiency of naturally sourced CESA catalyst was investigated by choosing
the reaction of 4-chlorobenzaldehyde 1b (1 mmol), malononitrile 2 (1 mmol) and β-naphthol 3a (1mmol)
as a model substrate for the synthesis of 3-amino-1-(4-chlorophenyl)-1H-benzo[f]chromene-2-carbonitrile
4b. First, we concentrated on determining the optimal reaction conditions, which included a variety of
solvents, amounts of synthesized CESA catalyst, and temperatures for model reaction. These were
examined for our proposed synthetic conversion, and those are summarized in table 1. Preliminary optimized reaction conditions indicated that, in the absence of the catalyst and solvent, even
under room temperature or at reflux conditions (Table 1, entry 1), only a trace amount of the desired
product 4b was obtained (Table 1, entries 1, 2). However, the model reaction was examined in the
absence of a catalyst, but in the presence of ethanol and water as solvents, only a small amount of the
desired product 4b was obtained at room temperature (Table 1, entries 3, 4). This result reveals that a
catalyst must be required for this transformation. We notice that, in the absence of solvents, the yield of
desired product 4b is improved due to the addition of the CESA catalytic amount, but there is no further
improvement in the yield of the desired product after adding the catalytic amount (Table 1, entry 5) and
even at reflux conditions (Table 1, entry 6). The result indicates that there is a necessity of solvent to
improve the yield of the desired product. Interestingly, when this model reaction was tested out in ethanol
as a solvent with varying catalytic amounts (2, 5, 10, or 12 wt.%) of CESA at room temperature, gave 85%
to 98 % corresponding product yield after 05 minutes (Table 1, entries 7-10). Page 6/21 Notably, it was found that the yield of the desired product drastically increased when increasing the
catalytic amount from 2 to 10 wt. % in the presence of EtOH (Table 1, entry 7-9). In contrast, with raising
the catalytic amount, there were no improvements in the yield of desired product 4b (Table 1, entry 10). It
was found that at reflux conditions, there were no improved effects on the product yield (Table 1, entry
11). Therefore, 10 wt. 3.2 Optimization of the reaction conditions In terms of green chemistry, the recyclability of the catalyst with high efficiency of the product yield of the
heterogeneous CESA catalyst are greatly desirable. Recyclability was the most important point for these
catalysts. Therefore, to prove the recyclability of the CESA catalyst, it was checked in the synthesis of 2-
amino-4H-benzochromene 4b using 4-chlorobenzaldehyde 1b, malononitrile 2 and β-naphthol 3a under
the optimized reaction conditions. After reaction completion, the reaction mixture was separated with ethyl acetate (4 X 20 mL) to obtain
product 4b without the use of column purification. The remaining aqueous phase of the CESA catalyst Page 7/21 Page 7/21 was dried in an oven for three hours at a temperature 120 o C and reused for the next run of the model
reaction under the same reaction conditions (scheme 2). The result presented in Fig. 9 demonstrates that
the yield of the desired product with each reuse of the catalyst is slightly reduced to a small extent. Fig. 9
shows that the isolated CESA catalyst from the reaction mixture was re-used for five successive runs
without significantly losing catalytic efficiency. A hot filtration test was used to detect the heterogeneity of the recovered CESA catalyst. The results
show that detectable methods with EDX (Fig. 6) have a similar composition, with K (40.21 %), Ca (33.72
%), Si (10.07 %), Na (5.37 %), Fe (5.81 %), Mg (1.83 %), Cl (2.51 %), and Mn (0.38 %). The catalytic activity of the CESA after five runs was confirmed by characterizing XRD, SEM, and EDX. As
shown in the XRD pattern of the CESA catalyst after 5th run (Fig. 10), the phase composition peaks of the
CESA, after 5 times of recovery, remain constant. Also, comparison to the SEM images results of recycled
CESA catalyst shows that, without any major significant change, CESA particles have slightly decreased
in sizes (Fig. 11). Moreover, the EDX spectra of the recycled CESA catalyst have been displayed in Fig. 12. According to the EDX pattern of CESA after the recovery test, suggesting the CESA has almost the same
phase composition of the elements in the catalyst, after consecutive 5 runs. The results shown in Fig. 10,
11, and 12 indicate that CESA has the characteristics of an excellent reusable catalyst. 3.2 Optimization of the reaction conditions According to the characteristics of the CESA catalyst and a literature survey, the proposed mechanism for
the synthesis of biologically active 2-amino-4H-benzochromene derivatives from one-pot tandem
Knoevenagel-Michael reaction of aryl aldehyde, malononitrile and diverse phenolic nucleophiles (β-
naphthol or α-naphthol) by the CESA catalyst is illustrated in Scheme 3. At first, the Knoevenagel reaction
occurs between aryl aldehyde and malononitrile; its methylene group is activated by the loss of an acidic
proton in the presence of a CESA catalyst, followed by the removal of a water molecule, which affords the
arylidinemalononitrile (I) as an intermediate. In the second step, this intermediate (I) reacts with phenolic
nucleophiles, β-naphthol via Michael addition, affording intermediate (II). Finally, an intramolecular
heterocyclization of the intermediate (III) followed by tautomerization afforded the formation of the
desired 2-amino-4H-benzochromene derivative (4). In order to show the efficiency and accessibility of the present work, we compared this method for the
synthesis of the desired product of model reactions 4b and 5b with some previously reported catalysts
(Table 3, entries 1 to 12 and 14 to 23). Each of the reported methodologies has its own merits, but some
of them suffer from drawbacks like harsh reaction conditions, longer reaction times, low yields, and the
use of expensive catalysts. The results summarized in Table 3 clearly show that the access of CESA
catalyst in the present methodology is more effective than reported methods. Page 8/21 Table 3 Comparison of the activity of CESA catalyst with reported catalyst for the synthesis of 4b and
5b. Naphthol
Entry
Reaction conditions
Time
(min.)
Yield
(%)
Ref. Conclusion As a conclusion, we have introduced an efficient one-pot multi-component procedure for the synthesis of
pharmaceutically active 2-amino-4H-benzochromene derivatives in the presence of the CESA as a
reusable natural catalyst in ethanol at room temperature. This method has the important advantages like
simplicity in operation, excellent yields in a short reaction time, using ethanol as a green solvent, and
avoids expensive catalyst. Moreover, the recovered catalyst can be successively reused in five runs
without a significant loss in catalytic activity. These are the key benefits of the present method. Utilization
of a natural waste material as a source of a catalyst not only provides an opportunity to catalyze organic
reactions but also solves the problems of the waste generated. Thus, this CESA catalyst is a better and
more practical alternative for green processes. 3.2 Optimization of the reaction conditions β-
Naphthol
1
Hexadecyl trimethyl ammonium bromide (HTMAB) (10
mol%)/H2O/Reflux
360
82
[22]
2
KF-Al2O3 (200mg)/Grinding
1440
84
[26]
3
Diazobicyclo[2.2.2]octane (30 mol%)/EtOH(5 mL)/RT
120-
240
72
[27]
4
Na2HPO4 (5 mol%)/120 oC
60
92
[29]
5
Fe(HSO4)3 (0.034 mg)/CH3CN (10 mL)/Reflux
240
85
[30]
6
Thiourea dioxide (10 mol%)/H2O(2 mL)/50 oC
480
87
[31]
7
Basic ionic liquid (20 mol%)/70 oC
03
90
[33]
8
Nano zeolite clinoptilolite (0.01 gm.)/H2O
(5mL)/Reflux
20
92
[34]
9
SiO2@Im-Fc[OAc] (10 mg)/90 oC
20
92
[40]
10
Fe3O4@PS‐Arg MNPs (0.07 g)/EtOH (5 mL)/Reflux
10
95
[41]
11
γ-Fe2O3@HAp@CPTMS@AT (0.02 g)/EtOH (2
mL)/Reflux
15
98
[42]
12
GO-Fc@Fe3O4 (10 mg)/100 oC
25
80
[43]
13
CESA
05
98
[ * ]
α-
Naphthol
14
Hexadecyl trimethyl ammonium bromide (HTMAB) (10
mol%)/H2O/Reflux
240
93
[22]
15
TiCl4 (10 mol%)/RT
10
91
[23]
16
KF-Al2O3 (200mg)/Grinding
960
90
[26]
17
Diazabicyclo[2.2.2]octane (30 mol%)/EtOH(5 mL)/RT
120-
240
82
[27]
18
Fe(HSO4)3 (0.034 mg)/ CH3CN (10 mL)/Reflux
180
94
[30] Table 3 Comparison of the activity of CESA catalyst with reported catalyst for the synthesis of 4b and
5b 20 Basic ionic liquid (20 mol%)/70 oC
21
Nano zeolite clinoptilolite (0.01 gm.)/H2O
(5mL)/Reflux
20
95
[34]
22
Potassium 2-oxoimidazolidine-1,3-diide (POImD) 90.08
g)/H2O/MW (900 W)
05
96
[38]
23
γ-Fe2O3@HAp@CPTMS@AT (0.02 g)/EtOH (2
mL)/Reflux
15
99
[42]
24
CESA (10 wt. %)/EtOH/RT
10
96
[ * ]
esent work Basic ionic liquid (20 mol%)/70 oC
21
Nano zeolite clinoptilolite (0.01 gm.)/H2O
(5mL)/Reflux
20
95
[34]
22
Potassium 2-oxoimidazolidine-1,3-diide (POImD) 90.08
g)/H2O/MW (900 W)
05
96
[38]
23
γ-Fe2O3@HAp@CPTMS@AT (0.02 g)/EtOH (2
mL)/Reflux
15
99
[42]
24
CESA (10 wt. %)/EtOH/RT
10
96
[ * ]
ent work *Present work *Present work Acknowledgements The authors are grateful to the Indian Institute of Science (IISC), Bangalore and Common Facility Center
(CFC), Shivaji University Kolhapur for spectral analysis. Competing interests Not applicable Ethical Approval Not applicable Authors' contributions SKS and SSP conceived the experiments and contributed equally. Availability of data and materials Not applicable Funding Not applicable SKS and SSP conceived the experiments and contributed equally. Page 10/21 SPP and SKS performed the collection field material, characterization of the catalyst and molecular
laboratory work. SKS and SSP designed and directed the study and also helped draft the manuscript. SKS and SSP designed and directed the study and also helped draft the manuscript. All authors gave final approval for publication. All authors gave final approval for publication. All authors gave final approval for publication. 1. Y. Gu, Green Chem. 14, 2091 (2012) 1. Y. Gu, Green Chem. 14, 2091 (2012) 2. H. Duan, D. Wang, Y. Li, Green Chem. Soc. Rev. 2015, <bvertical-align:super;>44</bvertical-
align:super;>, 5778 (2015) 3. P.T. Anastas, L.B. Bartlett, M.M. Kirchhoff, T.C. Williamson, Catal. Today 5 3. P.T. Anastas, L.B. Bartlett, M.M. Kirchhoff, T.C. Williamson, Catal. Today 55, 11 (2000)
4. M.B. Gawande, S.N. Shelke, R. Zboril, R.S. Varma, Acc. Chem. Res. 47, 1338 (2014) 4. M.B. Gawande, S.N. Shelke, R. Zboril, R.S. Varma, Acc. Chem. Res. 47, 1338 (2014) 5. R.A. Sheldon, Chem. Soc. Rev. 2012, 41, 1437–1451. 5. R.A. Sheldon, Chem. Soc. Rev. 2012, 41, 1437–1451. 6. A. Domling, W. Wang, K. Wang, Chem. Rev. 112, 3083 (2012) 7. H. Pellissier, Chem. Rev. 113, 442 (2013) 8. S. Brauch, S.S. Berkel, B. Westermann, Chem. Soc. Rev. 2013, <bvertical-align:super;>42</bvertical-
align:super;>,4948 (2013) 9. G. Brahmachari, B. Banerjee, ACS Sustain. Chem. Eng. 2, 411 (2014) 10. R. Pratap, V.Ram, Chem. Rev. 114, 10476 (2014) 11. F. Schmitta, M. Golda, M. Rothemunda, I.C. Andronacheb, B. Biersacka, R. Schoberta, T. Mueller, Eur. J. Med. Chem. <background-color:#FFCC66;buvertical-align:baseline;>163</background-
color:#FFCC66;buvertical-align:baseline;>, 160 (2019) 12. C.P. Dell, Curr. Med. Chem. 5, 179 (1998) 13. T.R. Wiernicki, J.S. Bean, C. Dell, A. Williams, D. Wood, R.F. Kauffman, J.P. Singh, J. Pharmacol. Exp. Ther.<bvertical-align:super;> </bvertical-align:super;>278, 1452 (1996) 14. A.A. Hussein, I. Barberena, T.L. Capson, T.A. Kursar, P.D. Coley, P.N. Solis, M.P.J. Gupta, Nat. Prod. 67,
451 (2004) 15. A. Rafinejad, A. Fallah-Tafti, R. Rakesh Tiwari, N. Shirazi, D. Mandal, A. Shafiee, K. Parang, A. Foroumadi, T. Akbarzadeh, DARU J Pharm. Sci. 20, 100 (2012) Page 11/21
16. A.H. Bedair, H.A. Emam, N.A. El-Hady, K.A.R. Ahmed, A.M. El-Agrody, Il Farmaco 56, 965 (2001) 17. H.F. Ashraf, A. El-Wahab, Pharmaceuticals 5, 745 (2012) 17. H.F. Ashraf, A. El-Wahab, Pharmaceuticals 5, 745 (2012) 18. C.W. Smith, J.M. Bailey, M.E.J. Billingham, S. Chandrasekhar, C.P. Dell, A.K. Harvey, C.A. Hicks, A.E. Kingston, G.N. Wishart, Bioorg. Med. Chem. Lett. 5, 2783 (1995) 19. J. Bloxam, C.P. Dell, C.W. Smith, Heterocycles 38, 399 (1994) 20. T.S. Jin, J.C. Xiao, S.J. Wang, T.S. Li, X. R. Song, Synlett 13, 2001 (2003) 21. R. Maggi, R. Ballini, G. Sartori, R. Sartorio, Tetrahedron Lett. 45, 2297 (2004) 22. T.S. Jin, J.S. Zhang, L.B. Liu, A.Q. Wang, T.S. Li, Synth. Commun. 36, 2009 (2006) 23. B.S. Kumar, N. Srinivasulu, R.H. Udupi, B. Rajitha, Y.T. Reddy, P.N. Reddy, P.S. Kumar, J. Heterocycl. Chem. 43, 1691 (2006) 24. A.Q. Zhang, M. Zhang, H.H. 1. Y. Gu, Green Chem. 14, 2091 (2012) Chen, J. Chen, H.Y. Chen, Synth. Commun. 37, 231 (2007) Zhang, M. Zhang, H.H. Chen, J. Chen, H.Y. Chen, Synth. Commun. 37, 231 (2 25. J.S. Yadav, B.V.S. Reddy, M.K. Gupta, I. Prathap, S.K. Pandey, Catal. Commun. 8, 2208 (2007) dav, B.V.S. Reddy, M.K. Gupta, I. Prathap, S.K. Pandey, Catal. Commun. 8, 22 26. T.S. Jin, H. Xie, Y P. Xue, T.S. Li, Asian J. Chem. 20, 5145 (2008) 27. S. Balalaie, S. Ramezanpour, M. Bararjanian, J.H. Gross, Synth. Commun 28. M.P. Surpur, S. Kshirsagar, S.D. Samant, Tetrahedron Lett. 50, 719 (2009) 29. X.Y. Meng, H.J. Wang, C.P. Wang, Z.H. Zhang, Synth. Commun. 41, 3477 (2011) 30. H. Eshghi, S. Damavandi, G.H. Zohuri, Synth. React. Inorg. Met-Org. Chem. 41, 1067 (2011) 31. S. Verma, S.L. Jain, Tetrahedron Lett. 53, 6055 (2012) 32. K.R. Desale, K.P. Nandre, S.L. Patil, Org. Commun. 5, 179 (2012) 33. H.R. Shaterian, M. Mohammadnia, Res. Chem. Intermed. 41, 1301 (2015) 34. S.M. Baghbanian, N. Rezaei, H. Tashakkorian, Green Chem. 15, 3446 (2013) 35. Y. Zhen, H. Lin, S. Wanga, M. Tao, RSC Adv. 4, 26122 (2014) 36. S.J. Kalita, N. Saikia, D.C. Deka, H. Mecadon, Res. Chem. Intermed. 42, 6863 (2016) 37. L. Kheirkhaha, M. Mamaghania, N.O. Mahmoodia, A. Yahyazadeha, S.S.M. Ziabari, J. Chem. Res. 41,
21 (2016) 38. L.Z. Fekri, M. Barazandehdoust, Polycycl. Aromat. Compd. 41, 1274 (2021) 38. L.Z. Fekri, M. Barazandehdoust, Polycycl. Aromat. Compd. 41, 1274 (2 39. Z. Zarnegar, J. Safari, New J. Chem. 40, 7986 (2016) 40. R. Teimuri-Mofrad, M. Gholamhosseini-Nazari, E. Payami, S. Esmati, Res. Chem. Intermed. 43, 7105
(2017) 41. M.K. Karkhah, H. Kefayati, S. Shariati, Organometal. Chem. 33, e5139 (2019) 41. M.K. Karkhah, H. Kefayati, S. Shariati, Organometal. Chem. 33, e5139 42. P. Jahanshahi, M. Mamaghani, React. Kinet. Mech. Catal. 130, 955 (2020 42. P. Jahanshahi, M. Mamaghani, React. Kinet. Mech. Catal. 130, 955 (2020) 43. S. Mozaffarnia, R. Teimuri–Mofrad, M.R. Rashidi, J. Iran. Chem. Soc. 18, 43. S. Mozaffarnia, R. Teimuri–Mofrad, M.R. Rashidi, J. Iran. Chem. Soc. 18, 1455 (2021) 44. P.R. Boruah, A.A. Ali, B. Saikia, D. Sarma, Green Chem. 17, 1442 (2015) 45. P.B. Hiremath and K. Kantharaju, ChemistrySelect 5, 1896 (2020) 46. J. Song, B. Zhou, H. Zhou, L. Wu, Q. Meng, Z. Liu, B. Han, Angew Chem. Int. Ed. 54, 9399 (2015) 46. J. Song, B. Zhou, H. Zhou, L. Wu, Q. Meng, Z. Liu, B. Han, Angew Chem. Scheme Scheme 1, 2 and 3 are available in supplementary section. Scheme 1, 2 and 3 are available in supplementary section. 1. Y. Gu, Green Chem. 14, 2091 (2012) I Page 12/21 Page 12/21 47. M. Satheesh, M. Pugazhvadivu, B. Prabu1, V. Gunasegaran, A. Manikandan, J. Nanosci. Nanotechnol. 19, 4123 (2019)
48. T.L. Ting, R.P. Jaya, N.A. Hassan, H. Yaacob, D.S. Jayanti, M.A.M. Ariffine, J. Teknologi. 78, 85 (2016)
49. A. Apasi, P.B. Madakson, D.S. Yawas, V.S. Aigbodion, Tribo in Industry 34, 36 (2012)
50. S.K. Shinde, S.A. Damate, S.T. Morbale, M.U. Patil, S.S. Patil, RSC Adv. 7, 7315 (2017)
51. S.K. Shinde, M.U. Patil, S. Damate, S.S. Patil, Res. Chem. Intermed. 44, 1775 (2018)
52. M.U. Patil, S.K. Shinde, S.P. Patil, S.S. Patil, Res. Chem. Intermed. 46, 4923 (2020)
53. A. Pawar, S. Gajare, A. Jagdale, S. Patil, W. Chandane, G. Rashinkar, S. Patil, Catal. Lett., 152, 1854
(2022)
54. S.P. Patil, S.K. Shinde, M.U. Patil, S.S. Patil, Res. Chem. Intermed. 48, 3589 (2022) 47. M. Satheesh, M. Pugazhvadivu, B. Prabu1, V. Gunasegaran, A. Manikandan, J. Nanosci. Nanotechnol. 19, 4123 (2019) 48. T.L. Ting, R.P. Jaya, N.A. Hassan, H. Yaacob, D.S. Jayanti, M.A.M. Ariffine, J. Teknologi. 78, 85 (2016) 49. A. Apasi, P.B. Madakson, D.S. Yawas, V.S. Aigbodion, Tribo in Industry 34 50. S.K. Shinde, S.A. Damate, S.T. Morbale, M.U. Patil, S.S. Patil, RSC Adv. 7, 7315 (2017) 51. S.K. Shinde, M.U. Patil, S. Damate, S.S. Patil, Res. Chem. Intermed. 44, 1775 (2018) 51. S.K. Shinde, M.U. Patil, S. Damate, S.S. Patil, Res. Chem. Intermed. 44, 17 52. M.U. Patil, S.K. Shinde, S.P. Patil, S.S. Patil, Res. Chem. Intermed. 46, 4923 (2020) 53. A. Pawar, S. Gajare, A. Jagdale, S. Patil, W. Chandane, G. Rashinkar, S. Patil, Catal. Lett., 152, 1854
(2022) 53. A. Pawar, S. Gajare, A. Jagdale, S. Patil, W. Chandane, G. Rashinkar, S. Patil, Catal. Lett., 152, 1854
(2022) 54. S.P. Patil, S.K. Shinde, M.U. Patil, S.S. Patil, Res. Chem. Intermed. 48, 3589 (2022) 54. S.P. Patil, S.K. Shinde, M.U. Patil, S.S. Patil, Res. Chem. Intermed. 48, 3589 (2022) Tables Table 1-2 are available in the Supplemental Files section. Table 1-2 are available in the Supplemental Files section. 47. M. Satheesh, M. Pugazhvadivu, B. Prabu1, V. Gunasegaran, A. Manikandan, J. Nanosci. Nanotechnol.
19, 4123 (2019) Figures Page 13/21 Figure 1
Some examples of biologically active 2-amino-4H-benzochromene compounds. Figure 1 Figure 1 Some examples of biologically active 2-amino-4H-benzochromene compounds. Page 14/21
Figure 2
Preparation of CESA Catalyst Figure 2 Figure 2 Figure 2 Preparation of CESA Catalyst Page 14/21 Figure 3
DSC-TGA analysis of Parent coconut endocarp Shell Ash (CESA) Figure 3 DSC-TGA analysis of Parent coconut endocarp Shell Ash (CESA) Page 15/21 Figure 4
FTIR analysis of CESA catalyst Page 16/21
Figure 4
FTIR analysis of CESA catalyst FTIR analysis of CESA catalyst Page 16/21 Figure 5 Figure 5 Figure 5 Page 17/21
Figure 6
EDX spectrum of CESA catalyst. Figure 6 EDX spectrum of CESA catalyst. EDX spectrum of CESA catalyst. Figure 7
BET analysis of CESA catalyst BET analysis of CESA catalyst Page 18/21
Figure 8
SEM images of CESA catalyst (a) Normal view, (b) Magnified view Figure 8 Figure 8 Page 18/21
SEM images of CESA catalyst (a) Normal view, (b) Magnified view Page 18/21
SEM images of CESA catalyst (a) Normal view, (b) Magnified view SEM images of CESA catalyst (a) Normal view, (b) Magnified view Page 18/21 Figure 9
The recyclability of the CESA in the synthesis of 4b under optimized conditions. The recyclability of the CESA in the synthesis of 4b under optimized conditions. Page 19/21 Figure 10
The XRD pattern of CESA catalyst after run five. Figure 10 Figure 10 The XRD pattern of CESA catalyst after run five. Figure 11
SEM images of recycled CESA catalyst (a) Normal view, (b) Magnified view. Figure 11 SEM images of recycled CESA catalyst (a) Normal view, (b) Magnified view. EM images of recycled CESA catalyst (a) Normal view, (b) Magnified view. Page 20/21 Figure 12
EDX spectrum of recycled CESA catalyst. Figure 12 EDX spectrum of recycled CESA catalyst. EDX spectrum of recycled CESA catalyst. Supplementary Files This is a list of supplementary files associated with this preprint. Click to download. SupportingInformation10size.docx Table12.docx Table12.docx floatimage1.png
Scheme01.png
Scheme02.png
Scheme03.png floatimage1.png Page 21/21
|
https://openalex.org/W2767791706
|
https://europepmc.org/articles/pmc5766353?pdf=render
|
English
| null |
Synchronous Uterine Metastases from Breast Cancer: Case Study and Literature Review
|
Curēus
| 2,017
|
cc-by
| 4,098
|
Synchronous Uterine Metastases from
Breast Cancer: Case Study and Literature
Review Aisha Akhtar , Atul Ratra , Yana Puckett , Abu Baker Sheikh , Catherine A. Ronaghan
1
2
3
4
5 1. Surgery, Texas Tech Health Sciences Center Lubbock 2. TTUHSC, Texas Tech University Health
Sciences Center 3. Department of Surgery, TTUHSC 4. Student, Texas Tech University Health Sciences
Center 5. Department of Surgery, Texas Tech University Health Sciences Center Corresponding author: Atul Ratra, dratulratra@gmail.com
Disclosures can be found in Additional Information at the end of the article Open Access Review
Article Open Access Review
Article Open Access Review
Article Abstract Breast cancer rarely metastasizes to the uterus. Here, we report two breast cancer patients with
synchronous metastases to the uterus. Case 1 highlights a 46-year-old female with invasive
ductal carcinoma who presented with a breast mass and was found to have uterine enlargement
on positron emission tomography (PET) scan. Biopsy revealed a metastatic 4 mm focus of
breast cancer in the background of endometrial hyperplasia. Case 2 reports a 62-year-old
postmenopausal female diagnosed with lobular carcinoma of the breast following an abnormal
screening mammogram. A routine pap smear necessitated further workup, revealing
simultaneous endometrial and cervical metastasis. Both patients did not have any gynecologic
symptoms and presented a diagnostic challenge. Categories: Obstetrics/Gynecology, Radiation Oncology, Oncology
Keywords: breast cancer, endometrial cancer, metastasis, extra-genital Received 07/12/2017
Review began 11/10/2017
Review ended 11/10/2017
Published 11/13/2017
© Copyright 2017
Akhtar et al. This is an open access
article distributed under the terms of
the Creative Commons Attribution
License CC-BY 3.0., which permits
unrestricted use, distribution, and
reproduction in any medium,
provided the original author and
source are credited. DOI: 10.7759/cureus.1840 How to cite this article
Akhtar A, Ratra A, Puckett Y, et al. (November 13, 2017) Synchronous Uterine Metastases from Breast
Cancer: Case Study and Literature Review. Cureus 9(11): e1840. DOI 10.7759/cureus.1840 Introduction And Background Breast cancer is the most common cancer worldwide and accounts for 29% of all cancers in
women. It is the second leading cause of cancer mortality among women and the leading cause
of cancer mortality among women 29-60 years of age [1]. Invasive ductal carcinoma (IDC) of the
breast and invasive lobular carcinoma (ILC) of the breast account for 75% and 15% of all cases
of breast cancer, respectively [2]. Six percent of breast cancer patients present with metastases and approximately 30% develop
metastases following definitive treatment [3-4]. ILC is more likely to have metastases at
presentation than IDC [5]. Although the most common sites of breast cancer
metastasis, lung/pleura, liver, bone, and brain, are similar in ductal as well as lobular
carcinoma, lobular carcinoma is more likely to metastasize to the gastrointestinal system,
gynecologic organs, peritoneum-retroperitoneum, adrenal glands, and bone marrow [6]. Received 07/12/2017
Review began 11/10/2017
Review ended 11/10/2017
Published 11/13/2017 © Copyright 2017
Akhtar et al. This is an open access
article distributed under the terms of
the Creative Commons Attribution
License CC-BY 3.0., which permits
unrestricted use, distribution, and
reproduction in any medium,
provided the original author and
source are credited. Metastases to genital organs from extra-genital cancer sites are extremely rare. When present,
ovaries are the most common site of metastasis due to peritoneal spread, accounting for 75.8%
of metastases to genital organs [7-8]. Metastases to genital organs other than the ovaries thus
pose a significant diagnostic challenge. Here, we are presenting two cases of breast cancer with
metastases to the endometrium. 2017 Akhtar et al. Cureus 9(11): e1840. DOI 10.7759/cureus.1840 Review A 42-year-old Hispanic woman with no known comorbidities presented to the emergency room
with a two-month history of a painful and enlarging right breast mass, extending to the right
axilla. Her review of systems was positive for anorexia and an unspecified amount of weight loss
over the last two months. Social history was negative for smoking, alcohol, or illicit drug use. Family history was negative for any cancer. Physical examination revealed a firm, tender, and
immobile mass in the right breast at the one o’ clock position, concerning for breast cancer. An
abdominal exam revealed an enlarged uterus approaching the xiphoid process. An ultrasound
examination revealed a mass at the 12 o’ clock position in the right breast, associated with a
skin thickening of 4.2 mm and axillary lymphadenopathy, the largest measuring 1.2 cm in the
long axis. There were two, right supraclavicular nodes measuring 8.5 mm and 9.7 mm,
respectively. A core needle biopsy of breast mass revealed a poorly differentiated IDC with
modified Scarf-Bloom-Richardson grade III. An immunohistochemical (IHC) analysis
demonstrated that the tumor was moderately positive for estrogen receptor (ER) at 15%,
strongly positive for progesterone receptor (PR) at 50%, Ki-67 unfavorable at 30%, and
HER2/NEU-positive by fluorescence in situ hybridization (FISH). The axillary lymph node
biopsy was also consistent with metastatic breast cancer. A magnetic resonance imaging (MRI) breast was obtained to rule out any malignancy in
the contralateral breast, which only demonstrated a 5.5 cm right breast mass (clinical stage
T4N2). The staging workup, including the bone scan and computed tomography (CT) scan of
the chest, were unremarkable. However, a positron emission tomography (PET) scan
demonstrated massive uterine enlargement with diffuse hypermetabolic activity and bilateral
hydronephrosis due to external compression (Figure 1). FIGURE 1: PET CT of patient shows a large mass in the pelvis
(white arrows)
PET: positron emission tomography; CT: computed tomography FIGURE 1: PET CT of patient shows a large mass in the pelvis
(white arrows) PET: positron emission tomography; CT: computed tomography PET: positron emission tomography; CT: computed tomography A gynecology service was consulted, which performed an endometrial biopsy. The 2017 Akhtar et al. Cureus 9(11): e1840. DOI 10.7759/cureus.1840 2 of 9 histopathology revealed a stromal adenocarcinoma, positive for GATA-3, pancytokeratin, and
ER on IHC, suggesting breast cancer metastasis. The endometrial biopsy was morphologically
similar to the specimen obtained from breast biopsy, affirming endometrial metastasis of
breast cancer. The patient was discussed during a multidisciplinary conference and recommended to undergo
systemic chemotherapy followed by surgery. She completed six cycles of docetaxel, carboplatin,
trastuzumab, and pertuzumab. She had a good clinical response with a barely palpable, vague
right breast mass and a decrease in uterine size to the level of the umbilicus. Restaging the PET
scan showed no hypermetabolic activity. She then underwent a total abdominal hysterectomy
and a bilateral salpingo-oophorectomy. Final pathology showed a small metastatic deposit to
the left ovary, but no evidence of residual metastases in the endometrium. Given the stage IV
disease at presentation, the patient was started on maintenance chemotherapy with
trastuzumab and pertuzumab. She continues to be on maintenance therapy for over one year
without any evidence of recurrence or progression. 2017 Akhtar et al. Cureus 9(11): e1840. DOI 10.7759/cureus.1840 Case 2 A 62-year-old Caucasian woman with a long-standing history of smoking presented after a
screening mammogram revealed a suspicious 14 mm mass in the superior aspect of the left
breast. Her previous mammogram was five years ago, which was normal. Family history was
positive for breast cancer in mother and sister. A physical examination revealed a 1.5 cm,
palpable, immobile, nontender mass at the 12 o’clock position in the left breast. An ultrasound
showed a 2.9 x 2.5 x 2.0 cm mass at the 12 o’ clock position of the left breast and right axillary
lymphadenopathy. Provided contralateral axillary lymphadenopathy, an MRI was obtained to
look for an occult right breast lesion, which showed an axillary tail mass of 1 x 2 cm with
suspicious lymph nodes in the left breast. A core needle biopsy of the left breast mass was
consistent with invasive lobular carcinoma along with fragments of a sclerosing papilloma. IHC
staining demonstrated that the tumor specimen was strongly positive for ER and PR at 90% and
50%, respectively, Ki-67 of 5%, and HER2/NEU negative by FISH. A staging CT scan of chest,
abdomen, and pelvis revealed a mass in the left breast as well as a right axillary tail mass and
an enlarged uterus. A biopsy of the right axillary tail mass and lymph node also showed
findings similar to those from the left breast mass. A routine pap smear necessitated further
workup, revealing simultaneous endometrial and cervical metastasis. Given the enlarged uterus
of 5.6 x 4.0 cm with an endometrium of 5 mm, a gynecology service was consulted and an
endometrial biopsy was obtained, demonstrating metastatic lobular carcinoma, also involving
the endocervix. This has been presented in Figure 2. 3 of 9 FIGURE 2: Ultrasound of the abdomen shows a large
intrauterine mass. IHC staining of the endometrial biopsy specimen was positive for pancytokeratin, vimentin,
ER, PR, and GATA-3 and negative for PAX-8. This has been presented in Figure 3. FIGURE 2: Ultrasound of the abdomen shows a large
intrauterine mass. IHC staining of the endometrial biopsy specimen was positive for pancytokeratin, vimentin,
ER, PR, and GATA-3 and negative for PAX-8. This has been presented in Figure 3. FIGURE 3: Endometrial biopsy revealed a tumor specimen with
morphological features compatible with metastatic lobular
carcinoma (arrows). FIGURE 2: Ultrasound of the abdomen shows a large
intrauterine mass. FIGURE 2: Ultrasound of the abdomen shows a large
intrauterine mass. IHC staining of the endometrial biopsy specimen was positive for pancytokeratin, vimentin,
ER, PR, and GATA-3 and negative for PAX-8. This has been presented in Figure 3. IHC staining of the endometrial biopsy specimen was positive for pancytokeratin, vimentin,
ER, PR, and GATA-3 and negative for PAX-8. This has been presented in Figure 3. IHC staining of the endometrial biopsy specimen was positive for pancytokeratin, vimentin,
ER, PR, and GATA-3 and negative for PAX-8. This has been presented in Figure 3. IHC staining of the endometrial biopsy specimen was positive for pancytokeratin, vimentin,
ER, PR, and GATA-3 and negative for PAX-8. This has been presented in Figure 3. FIGURE 3: Endometrial biopsy revealed a tumor specimen with
morphological features compatible with metastatic lobular
carcinoma (arrows). ,
,
g
p
g
FIGURE 3: Endometrial biopsy revealed a tumor specimen with
morphological features compatible with metastatic lobular
carcinoma (arrows). FIGURE 3: Endometrial biopsy revealed a tumor specimen with
morphological features compatible with metastatic lobular
carcinoma (arrows). FIGURE 3: Endometrial biopsy revealed a tumor specimen with
morphological features compatible with metastatic lobular
carcinoma (arrows). The patient was discussed during a multidisciplinary conference, and the consensus was that
the patient has a bilateral breast lobular carcinoma with metastases of the right breast
carcinoma to the right axillary lymph nodes as well as the endometrium. She was offered
hormonal and systemic chemotherapy with or without hysterectomy. The patient declined any The patient was discussed during a multidisciplinary conference, and the consensus was that
the patient has a bilateral breast lobular carcinoma with metastases of the right breast
carcinoma to the right axillary lymph nodes as well as the endometrium. She was offered
hormonal and systemic chemotherapy with or without hysterectomy. 2017 Akhtar et al. Cureus 9(11): e1840. DOI 10.7759/cureus.1840 further treatment and was lost to follow-up. further treatment and was lost to follow-up. further treatment and was lost to follow-up. The patient declined any 2017 Akhtar et al. Cureus 9(11): e1840. DOI 10.7759/cureus.1840 4 of 9 Discussion We have described two cases of breast cancer with metastases to the uterus. Case 1 was a
patient with IDC who presented with endometrial metastases. Case 2 was a patient with ILC
who presented with endometrial as well as endocervical metastases. These cases are unique due
to their unusual pattern of metastases involving only the endometrial lining and/or cervix. Metastasis to genital organs other than ovaries from extra-genital cancers is extremely
uncommon. Mazur et al. studied 325 patients with metastases to genital organs from all
nonhematologic cancers, and only identified seven extra-genital tumors with endometrial
metastases [8]. Only two of these had uterine metastases from breast cancer. Kumar et al. studied 63 cases of uterine metastases from a variety of extra-genital cancers,
primarily based on an autopsy examination [7]. Breast cancer accounted for approximately 43%
of these cases, followed by colon cancer (18%), stomach cancer (11%), and pancreatic cancer
(11%). Although metastases primarily involved the myometrium only (64%), 33% of patients
also had endometrial involvement. Only two patients had isolated endometrial involvement,
both of whom were detected on autopsy examination. This study suggests that uterine
metastases from extra-genital cancers occur in the context of disseminated disease. These
metastases usually stay overt given the short life expectancy thereafter. However, both patients
presented here had synchronous isolated uterine metastases from breast cancer. Isolated
uterine metastases at diagnosis presented a significant therapeutic challenge to us. Although
the disease was considered to be stage IV, surgical treatment of isolated metastases was
considered in both patients. Such an approach would definitely be controversial based on
current guidelines. However, one patient who underwent surgery is clinically disease-free until
recent follow-up. We conducted a literature search of all patients with uterine metastases of breast cancer,
utilizing PUBMED. We identified a total of 23 previously published cases, which are
summarized in Table 1, along with the two cases we have reported [9-23]. Publication
Age
Histology
ER,
PR,
Her2
Stage at
Dx
Met Sites
Synch
Presentation/Symptoms
1
Karvouni [9]
51
Ductal
ER+,
PR-
TxN1M0
Endometrium, Cervix, Liver,
Bone
No
Vaginal bleed
2
Hara [10]
44
Lobular
ER+,
PR-,
Her2-
T3aN1M0
Endometrium
No
Vaginal bleed
3
Arslan [11]
57
Ductal
ER+,
PR+,
Her2-
T1bN3aM0
Endometrium, Myometrium
No
Abd pain, distention
4
Erkanli [12]
63
Mixed
ER+,
PR+,
Her2+
T2N1M0
Endometrium
No
No symptoms, abnormal
TVUS, elevated CA15-3
ER-,
Endometrium, Myometrium, 2017 Akhtar et al. Cureus 9(11): e1840. Discussion DOI 10.7759/cureus.1840 5 of 9 2017 Akhtar et al. Cureus 9(11): e1840. DOI 10.7759/cureus.1840 22
Sullivan [27]
83
Ductal
ER+,
PR-
LN+
Endometrium
No
Uterine enlargement,
endometrial polyp
23
Aranda [28]
Lobular
N/A
Localized
Endometrium
No
Endometrial polyp
24
Our case #
1
42
Ductal
ER+,
PR+,
Her2-
IV
Endometrium
Yes
Breast mass
25
Our case #
2
62
Lobular
ER+,
PR+
IV
Endometrium
Yes
Breast mass ER+,
PR-
LN+
Endometrium
No
Uterine enlargement,
endometrial polyp
N/A
Localized
Endometrium
No
Endometrial polyp
ER+,
PR+,
Her2-
IV
Endometrium
Yes
Breast mass
ER+,
PR+
IV
Endometrium
Yes
Breast mass 22
Sullivan [27]
83
Ductal
23
Aranda [28]
Lobular
24
Our case #
1
42
Ductal
25
Our case #
2
62
Lobular TABLE 1: Literature review of all cases of breast cancer with uterine metastases
utilizing PUBMED
ER: estrogen receptor; PR: progesterone receptor; Her2: human epidermal growth factor receptor 2; TVUS: transvaginal
ultrasound; CA15-3: cancer antigen 15-3; CEA: carcinoembryonic antigen TABLE 1: Literature review of all cases of breast cancer with uterine metastases
utilizing PUBMED
ER: estrogen receptor; PR: progesterone receptor; Her2: human epidermal growth factor receptor 2; TVUS: transvaginal Out of 23, nine were ductal and 13 were lobular. The remaining three were mixed, apocrine,
and metaplastic. Fifteen were ER positive, 13 were PR positive, and eight cases were Her2-
negative. Only one case was reported with the synchronous presentation of metastatic breast
cancer with endometrial metastasis. Vaginal bleeding was the most common presenting
symptom. Only one patient presented with uterine enlargement. Out of 23, nine were ductal and 13 were lobular. The remaining three were mixed, apocrine,
and metaplastic. Fifteen were ER positive, 13 were PR positive, and eight cases were Her2-
negative. Only one case was reported with the synchronous presentation of metastatic breast
cancer with endometrial metastasis. Vaginal bleeding was the most common presenting
symptom. Only one patient presented with uterine enlargement. A majority of patients with uterine metastases were positive for hormone receptors. It is
unclear whether there is a relation between hormone positive status and risk of uterine
metastases. We know tamoxifen, commonly used for hormone receptor-positive breast cancer,
leads to endometrial hyperplasia. It is plausible that these endometrial changes and the
associated angiogenesis lead to a favorable microenvironment for the seeding of breast cancer
metastases. The uterine metastases of extra-genital cancer can present a diagnostic challenge for
physicians as well as pathologists. Conclusions In conclusion, synchronous endometrial metastases from breast cancer are extremely rare and
can pose a significant diagnostic and therapeutic challenge. In the setting of isolated uterine
metastases from breast cancer, surgery might be a viable option, especially if symptoms are
present. However, it is unknown if surgery in these cases would prolong survival. The long-term
follow-up of such patients is warranted. Even when appropriate workup is done, it may be difficult to
distinguish between uterine cancer and the uterine metastases of breast cancer. Both have a
glandular architecture on histopathology and both are likely to demonstrate positivity for
hormone receptors. Immunohistochemical stains like GATA3 and cytokeratin should be
employed in such cases, which have a sensitivity of 86% for breast cancer [29]. 5
Scopa [13]
50
Lobular
PR+,
Her2+
TxN3M0
Cervix, Ovaries, Fallopian
tubes
No
Vaginal bleed
6
Scopa [13]
81
Lobular
ER+,
PR+
T1N3M0
Endometrium, Myometrium,
Cervix
No
Vaginal bleed
7
Sinkre [14]
58
Metaplastic
ER-,
PR+,
Her2-
T2N0M0
Endometrium, Myometrium,
Ovary
No
Vaginal bleed
8
Giordano
[15]
72
Lobular
ER+,
PR+
T2N1M0
Endometrium, Myometrium,
Cervix, Ovary
No
Vaginal bleed
9
Giordano
[15]
77
Lobular
N/A
T2N1M0
Endometrium
No
Vaginal bleed
10
Kennebeck
[16]
71
Ductal
ER-,
PR-,
Her2-
T1N1M0
Endometrium, Cervix,
Vagina
No
No symptoms, elevated
CEA
11
Al-Brahim
[17]
53
Lobular
ER+,
PR-
T2N1M0
Endometrium
No
Endometrial polyp,
vaginal bleed
12
Ramalingam
[18]
59
Ductal
ER+,
PR+
IIIA
Endometrium, Urinary
bladder
No
Urinary frequency, pelvic
mass
13
Huo [19]
66
Ductal
ER-,
PR-,
Her2-
T2N0M0
Endometrium, Myometrium
No
No symptoms, elevated
CEA, abnormal TVUS
14
Binstock
[20]
43
Ductal
ER+,
PR+,
Her2 -
IIA
Endometrium, Myometrium,
Fallopian tubes, Ovaries,
Cervix, Bones
No
Vaginal bleed
15
Toyoshima
[21]
62
Lobular
ER+,
PR+,
Her2+
T2N1M0
Fibroids, Myometrium
No
Abdominal compression,
fibroids, elevated CEA,
and CA15-3
16
Bezpalko
[22]
47
Lobular
ER+,
PR+,
Her2-
IV
Endometrium, Bone, Bone
marrow, Gallbladder
Yes
Vaginal bleed, breast
edema and induration,
cholecystitis
17
Houghton
[23]
62
Lobular
N/A
T2N1M0
Endometrium
No
Endometrial polyp
18
Houghton
[23]
92
Lobular
N/A
Localized
Endometrium
No
Vaginal bleed,
endometrial polyp
19
Corley [24]
58
Ductal
N/A
N/A
Endometrium, Pleura,
Peritoneum, Ovaries
No
Endometrial polyp,
vaginal bleed
20
Alvarez [25]
69
Lobular
ER+,
PR+
T2N1M0
Endometrium, Bone
No
Endometrial polyp,
vaginal bleed,
21
Lambot [26]
70
Apocrine
ER+
LN+
T1cN1M0
Endometrium
Endometrial polyp,
vaginal bleed 2017 Akhtar et al. Cureus 9(11): e1840. DOI 10.7759/cureus.1840 6 of 9 support was received from any organization for the submitted work. Financial relationships:
All authors have declared that they have no financial relationships at present or within the
previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or
activities that could appear to have influenced the submitted work. Disclosures Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors
declare the following: Payment/services info: All authors have declared that no financial 7 of 9 2017 Akhtar et al. Cureus 9(11): e1840. DOI 10.7759/cureus.1840 References 1. Siegel RL, Miller KD, Jemal A: Cancer statistics, 2016. CA Cancer J Clin. 2016, 66:7–30. 10.3322/caac.21332 2. Li CI, Uribe DJ, Daling JR: Clinical characteristics of different histologic types of breast cancer . Br J Cancer. 2005, 93:1046–1052. 10.1038/sj.bjc.6602787 . Li CI, Uribe DJ, Daling JR: Clinical characteristics of different histologic types of breast cancer . 3. SEER cancer statistics review, 1975-2012. (2014). Accessed: April 2015:
https://seer.cancer.gov/archive/csr/1975_2012/. 4. Fowble BL, Solin LJ, Schultz DJ, Goodman RL: Ten year results of conservative surgery and
irradiation for stage I and II breast cancer. Int J Radiat Oncol Biol Phys. 1991, 21:269–277. 10.1016/0360-3016(91)90771-U 5. Di Meglio A, Freedman RA, Lin NU, et al.: Time trends in incidence rates and survival of newly
diagnosed stage IV breast cancer by tumor histology: a population-based analysis. Breast
Cancer Res Treat. 2016, 157:587–596. 10.1007/s10549-016-3845-5 6. Borst MJ, Ingold JA: Metastatic patterns of invasive lobular versus invasive ductal carcinoma
of the breast. Surgery. 1993, 114:637–641. 7. Kumar NB, Hart WR: Metastases to the uterine corpus from extragenital cancers. A
clinicopathologic study of 63 cases. Cancer. 1982, 50:2163-2169. 10.1002/1097- 0142(19821115)50:10<2163::AID-CNCR2820501032>3.0.CO;2-F 8. Mazur MT, Hsueh S, Gersell DJ: Metastases to the female genital tract. Analysis of 325 cases . Cancer. 1984, 53:1978-1984. 10.1002/1097-0142(19840501)53:9<1978::AID-
CNCR2820530929>3.0.CO;2-1 9. Karvouni E, Papakonstantinou K, Dimopoulou C: Abnormal uterine bleeding as a presentation
of metastatic breast disease in a patient with advanced breast cancer. Arch Gynecol Obstet. 2009, 279:199-201. 10.1007/s00404-008-0665-9 9. Karvouni E, Papakonstantinou K, Dimopoulou C: Abnormal uterine bleeding as a presentation
of metastatic breast disease in a patient with advanced breast cancer. Arch Gynecol Obstet. 2009, 279:199-201. 10.1007/s00404-008-0665-9 10. Hara F, Kiyoto S, Takabatake D, et al.: Endometrial metastasis from breast cancer during
adjuvant endocrine therapy. Case Rep Oncol. 2010, 3:137-141. 10.1159/000313921 10. Hara F, Kiyoto S, Takabatake D, et al.: Endometrial metastasis from breast cancer during
adjuvant endocrine therapy. Case Rep Oncol. 2010, 3:137-141. 10.1159/000313921 11. Arslan D, Tural D, Tatlı AM, Akar E, Uysal M, Erdoğan G: Isolated uterine metastasis of
invasive ductal carcinoma. Case Rep Oncol Med. 2013, 2013:793418. 10.1155/2013/793418 12. Erkanli S, Kayaselcuk F, Kuscu E, Bolat F, Sakalli H, Haberal A: Lobular carcinoma of the
breast metastatic to the uterus in a patient under adjuvant anastrozole therapy. Breast. 2006,
15:558-561. 10.1016/j.breast.2005.10.008 13. Scopa CD, Aletra C, Lifschitz-Mercer B, Czernobilsky B: Metastases of breast carcinoma to the
uterus. 2017 Akhtar et al. Cureus 9(11): e1840. DOI 10.7759/cureus.1840 References Report of two cases, one harboring a primary endometrioid carcinoma, with review of
the literature. Gynecol Oncol. 2005, 96:543–547. 10.1016/j.ygyno.2004.09.064 14. Sinkre P, Milchgrub S, Miller DS, Albores-Saavedra J, Hameed A: Uterine metastasis from a
heterologous metaplastic breast carcinoma simulating a primary uterine malignancy. Gynecol
Oncol. 2000, 77:216–218. 10.1006/gyno.1999.5712 15. Giordano G, Gnetti L, Ricci R, Meriso C, Melpignamo M: Metastatic extragenital neoplasms to
the uterus: a clinicopathologic study of four cases. Int J Gynecol Cancer. 2006, 16:433–438. 10.1111/j.1525-1438.2006.00235.x 16. Kennebeck CH, Alagoz T: Signet ring breast carcinoma metastases limited to the
endometrium and cervix. Gynecol Oncol. 1998, 71:461–464. 10.1006/gyno.1998.5126 16. Kennebeck CH, Alagoz T: Signet ring breast carcinoma metastases limited to the
endometrium and cervix. Gynecol Oncol. 1998, 71:461–464. 10.1006/gyno.1998.5126 17. Al-Brahim N, Elavathil LJ: Metastatic breast lobular carcinoma to tamoxifen-associated
endometrial polyp: case report and literature review. Ann Diagn Pathol. 2005, 9:166–168. 10.1016/j.anndiagpath.2005.03.003 18. Ramalingam P, Middleton LP, Tamboli P, Troncoso P, Silva EG, Ayala AG: Invasive
micropapillary carcinoma of the breast metastatic to the urinary bladder and endometrium:
diagnostic pitfalls and review of the literature of tumors with micropapillary features. Ann
Diagn Pathol. 2003, 7:112–119. 10.1053/adpa.2003.50015 19. Huo Z, Gao Y, Zuo W, Zheng G, Kong R: Metastases of basal-like breast invasive ductal 19. Huo Z, Gao Y, Zuo W, Zheng G, Kong R: Metastases of basal-like breast invasive ductal 2017 Akhtar et al. Cureus 9(11): e1840. DOI 10.7759/cureus.1840 8 of 9 2017 Akhtar et al. Cureus 9(11): e1840. DOI 10.7759/cureus.1840 carcinoma to the endometrium: a case report and review of the literature. Thorac Cancer. 2015, 6:548–552. 10.1111/1759-7714.12195 20. Binstock A, Smith AL, Olawaiye AB: Recurrent breast carcinoma presenting as
postmenopausal vaginal bleeding: a case report. Gynecol Oncol Rep. 2014, 10:38–40. 10.1016/j.gynor.2013.06.003 21. Toyoshima M, Iwahashi H, Shima T, et al.: Solitary uterine metastasis of invasive lobular
carcinoma after adjuvant endocrine therapy: a case report. J Med Case Rep. 2015, 9:47. 10.1186/s13256-014-0511-6 22. Bezpalko K, Mohamed MA, Mercer L, McCann M, Elghawy K, Wilson K: Concomitant
endometrial and gallbladder metastasis in advanced multiple metastatic invasive lobular
carcinoma of the breast: a rare case report. Int J Surg Case Rep. 2015, 14:141–145. 10.1016/j.ijscr.2015.07.036 23. Houghton JP, Ioffe OB, Silverberg SG, McGrady B, McCluggage WG: Metastatic breast lobular
carcinoma involving tamoxifen-associated endometrial polyps: report of two cases and review
of tamoxifen-associated polypoid uterine lesions. Mod Pathol. 2003, 16:395–398. 10.1097/01.mp.0000062655.62606.86 24. Corley D, Rowe J, Curtis MT, Hogan WM, Noumoff JS, Livolsi VA: Postmenopausal bleeding
from unusual endometrial polyps in women on chronic tamoxifen therapy. Obstet Gynecol. 1992, 79:111–116. 25. Alvarez C, Ortiz-Rey J, Estévez F, de la Fuente A: Metastatic lobular breast carcinoma to an
endometrial polyp diagnosed by hysteroscopic biopsy. Obstet Gynecol. 2003, 102:1149–1151. 10.1016/S0029-7844(03)00082-6 26. Lambot MA, Eddafali B, Simon P, Fayt I, Noël J-C: Metastasis from apocrine carcinoma of the
breast to an endometrial polyp. Virchows Arch. 2001, 438:517–518. 10.1007/s004280000379 27. Sullivan LG, Sullivan JL, Fairey WF: Breast carcinoma metastatic to endometrial polyp . Gynecol Oncol. 1990, 39:96–98. 10.1016/0090-8258(90)90408-D 28. Aranda FI, Laforga JB, Martinez MA: Metastasis from breast lobular carcinoma to an
endometrial polyp. Report of a case with immunohistochemical study. Acta Obstet Gynecol
Scand. 1993, 72:585-587. 10.3109/00016349309058170 29. Braxton DR, Cohen C, Siddiqui MT: Utility of GATA3 immunohistochemistry for diagnosis of
metastatic breast carcinoma in cytology specimens. Diagn Cytopathol. 2015, 43:271-277. 10.1002/dc.23206 9 of 9
|
https://openalex.org/W3216269838
|
https://zenodo.org/record/5725000/files/Frontiers%20Output%204%20-%20Evaluation%20%26%20Dissemination%20report.pdf
|
English
| null |
FRONTIERS Intellectual Output 4
|
Zenodo (CERN European Organization for Nuclear Research)
| 2,021
|
cc-by
| 29,419
|
Introduction: The FRONTIERS consortium believe that the science classroom should provide challenging, open,
authentic and higher-order learning experiences for students. There should be opportunities for
students to participate in scientific practices and tasks, to use the discourse of science and work with
scientific tools. Key to stimulating students and discovering the next generation of frontier science
innovators is presenting frontier scientific ideas and activities that are closely related to new
technological achievements and everyday life. Under the unique circumstance of the Covid 19 pandemic, the consortium has been dynamic and
innovative in delivering the project outcomes whilst overcoming unparalleled challenges. Despite
the global lockdown of society, including in-person teaching & learning, the consortium has
delivered high-quality teaching resources that blend frontier science with pedagogical thought,
provided teacher training, facilitated virtual tours of research institutions to an international
audience of teacher and students, and built a community of educators passionate about frontier
physics. The FRONTIERS project designed, developed, and evaluated 21 demonstrators. This series of
innovative educational activities offer unique scientific resources to teachers and students organised
under a systematic pedagogical framework. Teachers are supported on an ongoing basis through the
FRONTIERS Community Support Environment, the online infrastructure that allows engagement and
interaction between teachers, research institutes and project organisers. A series of international e-schools, virtual visits, science contests, and teachers training activities
were organised as part of the FRONTIERS project. Hundreds of teachers got involved in the design
and development of innovative classroom activities in a collaborative way. Through the Frontiers
network teachers collaborated with the outreach teams of large-scale research infrastructures. Being part of a professional network encouraged interaction and provided opportunities to enrich
practices and professional context through cooperation within and between schools, universities,
and frontier research institutions. A series of virtual visits were organised for students where they were introduced to gravitational
wave astronomy and guided by expert scientists in a remote tour to Virgo, Europe’s advanced
gravitational wave astronomy facility. This gave students a chance to witness first-hand how
experiments are performed and how data acquired by these experiments are studied. This way of
introducing science helps students overcome the idea of it being complex and too difficult for them
to understand and helps them to see it as a tool to explore and understand nature. Introduction: The FRONTIERS Tool-Kit, “Effective Ways of Introducing Frontier Science in Schools”, was developed
and will be made available through the e-Twinning collaboration space to all European schools. The
Tool Kit provides a set of recommendation for a pan European roadmap for the introduction of
relevant activities in schools and how scientific work can be used to provide an engaging educational
experience through the exploration of “real science”. Furthermore, by bringing together researchers from different fields of Science, including particle
physics, astronomy and cosmology, this project promoted cooperation between managers of
different infrastructures to achieve trans-national collaboration and the designing of effective and
interdisciplinary educational activities. The project partners are proud to present this evaluation of the Frontiers Project outputs. 2 2 Conclusion: 3 3 The FRONTIERS project aimed to demonstrate how Nobel Prize winning science can be
systematically integrated in the school curriculum. This is demonstrated through a monitored-for-
impact use of innovative educational activities, the FRONTIERS Demonstrators, which provided
feedback for the take-up of such interventions. The project consortium proposed the main ways that
frontier science experiments can leverage educational activities are by: a) Making scientific and educational resources available worldwide through advanced
repositories and effective search mechanisms. b) Supporting effective community building between researchers, teachers and students. c) Creating virtual global classrooms by interconnecting the schools and the research ce The aim of the project was to mobilize 1,000 teachers and 10,000 students in the framework of the
proposed activities. Building on the above ways that frontier science experiments could leverage
educational activities, five project outputs were proposed as a means of achieving the main project
objects. The aim of the project was to mobilize 1,000 teachers and 10,000 students in the framework of the
proposed activities. Building on the above ways that frontier science experiments could leverage
educational activities, five project outputs were proposed as a means of achieving the main project
objects. 1. Select a series of scientific research outreach programmes that successfully introduce the
scientific methodology in school science education, by utilizing existing research infrastructures
of frontier research institutions enriched with online tools (data analysis tools, simulations &
games) and web-interactive educational material (O1). 2. Integrate these initiatives under a common educational approach and develop the FRONTIERS
Demonstrators that could be exploited and widely used from the educational communities in
Europe and beyond (O2). 2. Integrate these initiatives under a common educational approach and develop the FRONTIERS
Demonstrators that could be exploited and widely used from the educational communities in
Europe and beyond (O2). 3. Create virtual learning communities of educators, students and researchers and involve them in
extended episodes of playful learning. The proposed project will involve teachers, students and
researchers in collaborative learning activities. The development of the virtual learning
community will be enhanced by the FRONTIERS Community Support Environment (O3). 4. Systematic validate the proposed approaches and activities in order to identify their impact in
terms of the effectiveness and efficiency. The project will be implemented in schools, science
teacher training centres, and research centres in different countries and a detailed evaluation
report will be prepared (O4). 5. Conclusion: Design and implement a systematic raising awareness strategy that will contribute to the
effective communication of the project’s results and outcomes. A devoted Tool Kit (O5) will be
developed that will be uploaded to the eTwinning collaboration space to act as a starting point
for numerous collaborative projects between schools. Impact of Covid 19 The FRONTIERS project set out to empower science teachers to affect change by enabling them to
bring frontiers science into the classroom using an inquiry-based learning approach. This approach
was adopted to increase students’ interest and attainment in science. A major challenge arising during the lifetime of the project was the Covid-19 pandemic. While the
project achieved and even exceeded its stated objectives, the unprecedented societal lockdowns
prevented elements of the proposed project activities from continuing as originally envisaged. 4 Instead, project partners successfully delivered project objectives through a range of dynamic and
bespoke technical solutions. Instead, project partners successfully delivered project objectives through a range of dynamic and
bespoke technical solutions. A project objective was to provide face-to-face teacher professional development or multiplier
events and practice reflection meetings at a local level in the different partner countries. These
events took place face-to-face in 2019. Due to Covid-19 the project consortium could not continue
with face-to-face events. In March 2020 a decision was made to continue offering the learning and
teaching events using the online technologies available in the individual partner institutions or
supported by project partners. An evaluation of the face-to-face and online professional
development events was carried out using quantitative and qualitative methods. Feedback on the
Demonstrators was also gleaned from the teachers and this led to further refinement and
improvement of the Demonstrators ahead of final implementation. The project planned to either map the process for the effective implementation of frontier science in
school environments by integrating the Demonstrators into school practice or by extra-curriculum
activities (e.g. masterclasses, summer schools, contests). A consequence of the Covid-19 pandemic
and resulting societal lockdowns, was the closure of schools to students for long periods. This
resulted in distance learning being adopted and heavily reduced in-person contact times on return. With normal laboratory-based classes not occurring, project partners decided that in-school
implementation of FRONTIERS Demonstrators, was an unrealistic and unreasonable expectation of
teachers and school authorities. Alternatively, priority was given to direct engagement and
empowerment of teachers to implement the Demonstrators, through international e-school events. The project partners also focused on supporting teachers to engage students in a meaningful way
through facilitated virtual visits to large research infrastructure such as CERN, Virgo and Pierre
Auger. Project Impact The FRONTIERS approach aims to foster a culture of cooperation between research infrastructures
and schools. By spreading good practices between outreach groups of large-scale research
infrastructures and encouraging them to develop their activities in complementary ways the
consortium aimed to demonstrate a vision of the science classroom of tomorrow. As far as the
target groups are concerned the project is focusing on both students and teachers. The FRONTIERS project aims to attract young people to science and pool talent to scientific careers - By simulating the work of the researcher in the classroom - By promoting a better understanding of how science works - By enhancing students’ science related career aspirations - By promoting inquiry-based science teaching and learning Key to achieving project aims is to empower science teachers to affect change. The consortium
believe that the above actions can only be accomplished with the full collaboration and engagement
of teachers and their schools. With this in mind, the FRONTIERS project set out to expand opportunities for teachers’ professional
development, including occasions to interact with working scientists, to help stimulate and innovate
their science classroom, and more generally think differently about their students’ learning of and
about science. By offering teachers a large repertoire of tools and applications, along with a detailed school-based
framework for their effective introduction, the FRONTIERS project sought to empower teachers. This
is not only meant to change their teaching practice and introduce contemporary scientific issues in
their lessons, but also to propose and initiate the necessary changes in their schools, to allow for a
more seamless introduction of ICT innovations. The challenge of professional development for science teachers is to create optimal collaborative
learning situations in which the best sources of expertise are linked with the experiences and current
needs of the teachers. Evaluation Plan The project evaluation section of this report provides an overview of the evaluation process
employed. This overview starts by revisiting the project aims and objectives, the delivered
intellectual outputs and details of their intended impact. The section finishes with an account of the
methodology employed to evaluate the effectiveness of the Frontier project deliverables. Impact of Covid 19 Due to the covid-19 situation the one-week international summer schools planned for July 2020 and
July 2021 and the one-week international winter school in February 2021 were redesigned to take
place fully online. The planned one-week winter school took place over two consecutive weekends
at the end of January and beginning of February and there were individual meetings with teachers
between both weekends. The summer schools took place as fully online one-week schools. Teachers were supported through online learning tools to participate in master classes, summer and
winter schools, teacher networks and contests from March 2020 until July 2021. It is evident from
the teacher feedback that the organisation and structure of these online events provided space for
them to draw on the expertise of the project consortium and collaborate with teachers to gain a
sound understanding of the Demonstrators and of the topics of Frontier science. The alternative approach adopted by the project partners in a time of profound instability and
change, facilitated the stated objectives of the FRONTIERS Project. Teachers were involved in the
design and development of classroom activities in a collaborative way. An international professional
community was established, allowing collaboration with not just other teachers, but also with the
outreach teams of large-scale research infrastructures and universities. As documented in the report
this objective exceeded our expectation as teachers created classroom activities based on the topics
of the FRONTIERS Demonstrators. The teacher created resources are available on the Frontiers
website and can be accessed by teachers across Europe and beyond. 5 5 5 Students The student target group of the project are aged 12-18 with a focus on the development of their
content and concept knowledge, skills and attitudes. The project evaluation will mainly focus on
estimating the impact of the project overall activities on the following distinct areas: a. students’ ability to operate experimental techniques to conduct a scientific inquiry b. students’ ability to monitor, record and analyse data y
,
y
c. enhancement of students’ problem-solving competence and interdisciplinary thinking
d
enhancement of a positive attitude towards STEM disciplines c. enhancement of students’ problem-solving competence and interdisciplinary thinkin d. enhancement of a positive attitude towards STEM disciplines Institutions and Schools At institutional and school level, the project aims to assess and estimate impact at the following
areas: a. to what extent school authority encouraged or facilitated participation of its staff
teachers in training and teacher development programmes b. to what extent school authority encouraged or facilitated development of
interdisciplinary educational activities e.g. by altering school’s timetable so that teachers
from different STEM disciplines can work together c. to what extent schools collaborated or plan to collaborate in the future with other
schools in their area/region/country or other countries d. to what extent the participation of the schools in the FRONTIERS project was considered
as recognition or distinction of achievement and further facilitated the participation in
other projects and initiatives Teachers Teachers
With respect to participant teachers the project aimed to assess and estimate impact in the
following areas: a. the effectiveness of the training seminars and workshops b. the extent participant teachers developed interdisciplinary educational activities c. the extent participant teachers collaborated with other teachers effective integration of already established strong educational communities with project consortium
developed online resources and activities. effective integration of already established strong educational communities with project consortium
developed online resources and activities. Measuring of Impact This evaluation report aims to measure the impact and the effectiveness, both quantitatively and
qualitatively, of the project at three levels; at teacher, student and institutional/school level. The aim of the project was to mobilize 1,000 teachers and 10,000 students in the framework of the
proposed activities. The development of the FRONTIERS users’ community was based on the 6 6 6 Methodology gy
Given the broad range of project aims, intended impacts and participants involved, a mixed methods
evaluation approach was adopted. Utilising both quantitative and qualitative data collection and
analysis, partners set out to systematically and rigorously evaluate the impact of the project outputs
have on integrating Nobel prize winning science into the classroom. The evaluation sets out to
determine whether the project had achieved its intended impact and how it might be improved. Multiple data collection methods and analysis were developed to evaluate the effectiveness of
project outputs. To harness the multiple perspectives of relevant project stakeholders, a series of
instruments and evaluation tools were developed to be more effective and user-friendly than the 7 7 7 7 traditional. The expertise of the project partners has allowed for the development of a dynamic set
of evaluation tools through quantitative and qualitative methods. A mixed methods approach was adopted by project partners to evaluate the multiple stages and
perspectives relevant to the FRONTIERS project outputs. According to Johnson, Onwuegbuzie and
Turner (2007), the combining of elements of qualitative and quantitative research approaches in one
study, has the benefit of providing breadth and depth of understanding and corroboration. Whilst
gaining valuable insight into the lived experience of those closest to the phenomenon through the
qualitative phase, the inclusion of the quantitative methods can make the study findings more
generalisable and acceptable to quantitative-biased audiences. Developing our evaluation plan for the FRONTIERS project, it was decided to employ a Convergent
Mixed Methods Evaluation Design, as described by Creswell and Plano Clark (2018). The Convergent
Mixed Methods Evaluation Design is appropriate for complex projects with multiple stages and
objectives over several years. It allows for the evaluation of outcomes and activities undertaken in
the cultural context with researchers undertaking several iterations of data collection, analysis,
integration and implementation. Onwuegbuzie and Hitchcock (2017) advocate conducting mixed methods impact evaluations of
educational programmes, as the combination of qualitative and qualitative methods provides a
rigorous impact evaluation that yields strong inferences with a focus on if and how the programmes
have impacted society. While the quantitative approaches permit generalisation and test statistical
difference, Bamberger, Vijayendra and Woolcock (2010) explain, the qualitative approaches allow in-
depth description, process analysis and patterns of social interaction. Methodology It is also argued that a mixed
method orientation to evaluation is critical to the three dimensions of transferability, generalisation
and sustainability of evaluating an educational programme, as a “singular commitment to one
paradigm might yield somewhat impoverished capacity to think about these concerns”
(Onweugbuzie and Hitchcock, 2017, p. 59). 8 8 8 Evaluation Summary The FRONTIERS project aimed to demonstrate how Nobel Prize winning science can be
systematically integrated in the school curriculum. By fostering a culture of cooperation to spread
Through the project activities and outputs the consortium sought to bring Nobel prize winning
frontier physics to the classroom by: a) Making scientific and educational resources available worldwide through advanced
repositories and effective search mechanisms. a) Making scientific and educational resources available worldwide through advanced a) Making scientific and educational resources ava
repositories and effective search mechanisms. a) Making scientific and educational resources ava
repositories and effective search mechanisms. repositories and effective search mechanisms. b) Supporting effective community building between researchers, teachers and students. b) Supporting effective community building between researchers, teachers and students. c) Creating virtual global classrooms by interconnecting the schools and the research centres. c) Creating virtual global classrooms by interconnecting the schools and the research centr Key to achieving the project aim is to empower science teachers to affect change. The consortium
believe that the above actions can only be accomplished with the full collaboration and engagement
of teachers and their schools. The Frontier project expanded the opportunities for teachers’
professional development, providing occasions to interact with working scientists, helping them to
introduce innovations in their science classroom, and think differently and more innovatively about
their students’ learning. Project Impact The evaluation report measured the impact and effectiveness of the FRONTIERS project outputs in
bringing Nobel prize winning physics to the classroom. The impact was measured both qualitatively
and quantitatively through a convergent mixed methods impact evaluation approach. This mixed
methods approach allowed for the evaluation of outcomes and activities undertaken in the cultural
context with researchers undertaking several iterations of data collection, analysis, integration and
implementation. Onwuegbuzie and Hitchcock (2017) advocate conducting mixed methods impact evaluations of
educational programmes, as the combination of qualitative and qualitative methods provides a
rigorous impact evaluation that yields strong inferences with a focus on if and how the programmes
have impacted society. This evaluation of the FRONTIERS project set out to measure the impact and effectiveness, both
qualitatively and quantitatively, of the project at teacher, student and institutional level. The impact
and effectiveness of the project at these three levels will be discussed below. Mobilisation Target The first key performance indicator of the FRONTIERS project was to mobilise 1,000 teachers within
the framework of the project. It has been demonstrated, through the range of activities and
platforms employed by the project to engage and mobilise teachers, the consortium has mobilised
well in excess of the target figure. Project activities that were designed to engage and mobilise teachers included the project multiplier
events and online community support. The multiplier events held by the project included the Initial
Teacher Training, International e-Schools and Virtual Visits. The range of platforms used to engage
teachers through the online community support involved Facebook, project website, OSOS platform
and Google Classroom. Teacher Mobilisation Figures
Synchronous Engagement
Initial Teacher Training
303
International e-Schools
277
Virtual Visits
192
Masterclasses
71
Total
843
Asynchronous Engagement
Facebook
1,111
Project Website
8,305
OSOS Platform
174
Google Classroom
70
Total
9,660 Teachers Key to the success of the FRONTIERS project was empowering teachers to affect change and bring
prize winning physics to the classroom. The consortium believed that the project aims can only be
accomplished with the full collaboration and engagement of teachers and their schools. As a result,
the Frontier project set out to expand opportunities for teachers’ professional development. In addition to the stated mobilisation target of 1,000 teachers, the project also aimed to assess and
estimate impact at the following areas: a) Effectiveness of the training seminars and workshops a) Effectiveness of the training seminars and workshops b) Extent that participant teachers developed interdisciplinary educational activities c) Extent that participant teachers collaborated with other teachers 9 9 levels of teacher confidence and ability to motivate students. These quantitative findings were also
supported through the qualitative findings from focus group and round table discussion with
participants. levels of teacher confidence and ability to motivate students. These quantitative findings were also
supported through the qualitative findings from focus group and round table discussion with
participants. It was widely acknowledged that the “hands-on” and practical approach of the training workshops
motivated teachers to participate more fully and enthusiastically. The inquiry-based learning
approach adopted by the project was noted as being of great interest and benefit to teachers and
students. The demonstrators in their content and design facilitated bringing the scientific approach
to the classroom in both its content and pedagogical approach. The age and experience of the teachers taking part in the training were important variables in
relation to the impact the training had on the participants. A statistically significant relationship was
found between participant age and the reported increase in confidence. While all age groups
reported an increase in confidence in delivering Nobel prize winning physics, participants aged 34
and under had the highest average increase with more than 2 points on a 10-point scale. Survey data collected from the International e-School participants found a significant increase was
recorded in the average Self-Efficacy score from pre (M=3.8) and post (M=4.5) results on a 5-point
scale. The confidence of the participant teachers with content knowledge of modern physics
increased comparing pre (M=3.2) and post (M=4.0) training scores. The post training, participants
reported feeling competent to design educational content relating to modern physics (M=4.2, SD
0.635). The feeling that the FRONTIERS Demonstrators and International e-Schools would provide teachers
with the tools, knowledge and confidence to bring frontier science to the classroom was held
amongst participants. Following the completion of the International e-School, participants reported
being very interested in implementing the FRONTIERS Demonstrators with their students (M=4.82,
SD=0.39). Effectiveness of training seminars and workshops The FRONTIERS project delivered a series of high-quality training seminars and workshops to science
teachers, within the partner countries and beyond. Central to the effectiveness of these training
events was the development of the 21 FRONTIERS Demonstrators. The purpose of these outputs was
to empower science teachers in bringing Nobel prize winning physics to the classroom. The selection and development of the Demonstrators, employing quality criteria and pedagogical
framework selection consideration, ensured high quality resources to support bringing Nobel prize
winning physics to the classroom. The 15 initial teacher training events held across the project partner countries provided feedback on
the quality and relevance of the demonstrators, and also the effectiveness of the training provided
to teachers on how to effectively employ the resources in the classroom. The feedback from the 303
participants of the initial teacher training informed the development of the International e-School
series. Using a 5-point scale, questionnaire responses indicated participants were interested and
enjoyed the International e-Schools (M=4.25, SD=0.698). Participants also reported the e-School
content was very valuable and useful to them (M=4.8, SD=0.48). Key to measuring the effectiveness of the training events provided by the FRONTIERS project, was
measuring the impact the training and resources had on the self-efficacy of the participating
teachers. The evaluation methods used to measure the impact and effectiveness of project training
events showed an increase in teacher confidence with the subject of Nobel prize winning physics, up
21%, as well as their ability to motivate students to learn about the topic, up 8%. Analysis of
questionnaire data showed a statistically significant relationship between pre and post training 10 Extent that participant teachers developed interdisciplinary educational activities The FRONTIERS project developed 21 Demonstrators to aid teachers in bringing Nobel Prize winning
physics to the classroom. The range of physics categories covered through the Demonstrators and
the training provided to participants, facilitated the teachers to engage with and develop a range of
interdisciplinary educational activities in the field of physics. The Demonstrators were organised according to categories of frontier physics, from basic physics to
astronomy, cosmology and high energy physics. This interdisciplinary repository of resources and
training were designed to encourage participant teachers to upskill and expand their teaching into
other branches of physics that are less frequently delivered. A key feature of the International e-School was establishing working groups of participating teacher
to work collaboratively to create their own interdisciplinary resources based the FRONTIERS
Demonstrators. This working group approach, supported by project facilitators and frontier physics
researchers, was considered instrumental in participants reporting feeling competent to design
educational content relating to modern physics in post training survey data (M=4.2, SD 0.635). From the focus group findings, Teacher E explained that working with other colleagues to develop
resources and “meeting with mentors was very helpful to strengthen our confidence to talk to our
own students about these topics…” This supported development of professional confidence and self- 11 11 11 efficacy was felt would be of great benefit to the teachers when delivering Nobel prize winning
physics content in their classrooms. The results of these interdisciplinary collaborations are hosted on the FRONTIERS website, as
resources to be shared with other interested and motivated science teachers looking for inspiration. Students The student target group of the project was aged 12-18 years. The focus of the project was on the
development of student content and concept knowledge and support the development of skills and
attitudes towards frontier physics. Extent that participant teachers collaborated with other teachers Highlighted through feedback from the initial teacher training events, a priority for the International
e-Schools was to promote and facilitate participating teachers to work collaboratively with fellow
professionals to explore the resources. The success of this was highlighted in the project focus group
findings with participants. The focus group participants highlighted that the collaborative design of
the Winter School helped the teacher develop a deeper understanding of the content and
educational tool by allowing the professionals to explore and develop the resources together. Survey data findings showed participants were satisfied with the level of collaboration with other
teachers (M=4.18, SD=1.01), and very satisfied with the level of collaboration they experienced with
course organisers (M=4.68, SD=0.72). In focus group discussion that reflected the general feeling of
participants, Teacher C stated, “I liked all things…but if had to choose one thing it would be the
collaboration with colleagues far away from me.” As the International e-Schools were held virtually, online collaborative tools had to be used to
facilitate the teacher engaging with the content and each other. Several of the collaborative tools
employed by the Frontier Project were subject to praise. The use of Zoom to host the live events and
sharing of resources by email was functional and accessible to all participants. In the working groups,
Google Slides received considerable praise. In additional to working collaboratively with other participating teacher at the International e-
School, the FRONTIERS project also set out to facilitate teachers working collaboratively with other
professionals through the online community support and e-twinning projects. International e-School
participants were very interested in collaborating with colleagues to create e-twinning projects
(M=4.64, SD=0.73). The appetite of teachers to collaborate with other professional internationally
was summed by Teacher C in the focus group discussion, who said, ““The main reason I wanted to be
part of the e-Winter school was it was very interesting and exciting to collaborate and to listen to
what teachers from other countries had to say.” Student Skills Development Through the collection of qualitative data from International e-School participants, teachers
highlighted that the Demonstrator resources facilitated students working with real-life data using
tools and techniques reflecting the work of physicists in the field. Despite the barriers presented by
the Covid-19 pandemic and subsequent societal lockdowns, the FRONTIERS Demonstrators has been
shown to be effective in promoting the prioritised student impact criteria as set out below. a. students’ ability to operate experimental techniques to conduct a scientific inquiry a. students’ ability to operate experimental techniques to conduct a scientific inquiry b. students’ ability to monitor, record and analyse data c. enhancement of students’ problem-solving competence and interdisciplinary thinking
d. enhancement of a positive attitude towards STEM disciplines c. enhancement of students’ problem-solving competence and interdisciplinary thinkin p
g
p
p
y
g
d. enhancement of a positive attitude towards STEM disciplines d. enhancement of a positive attitude towards STEM disciplines Integral to the development of the FRONTIERS Demonstrators was the inquiry-based pedagogical
approach. Feedback from teachers that participating in the International e-School training events
highlighted the approach was of great interest and benefit to both teachers and students. The
internal evaluation criteria for development of the Demonstrators included the incorporation of
student skills development. These included: Integral to the development of the FRONTIERS Demonstrators was the inquiry-based pedagogical
approach. Feedback from teachers that participating in the International e-School training events
highlighted the approach was of great interest and benefit to both teachers and students. The
internal evaluation criteria for development of the Demonstrators included the incorporation of
student skills development. These included: •
Learning and Innovation skills include communication, collaboration, problem solving and
thinking outside the box. •
Literacy skills involves the use of technology, ability to read statistics and graphs, access,
evaluate and create media. •
Life skills is a term used to describe the ability to set goals, plan use of resources, respond to
feedback, take responsibility for collecting, organizing and analysing data. Through the voice of the teachers who undertook the training to implement the FRONTIERS
Demonstrators in the classroom, the value and effectiveness of the resource in promoting student
skills development was evident. teachers, it can be stated that the project also achieved its stated aim of mobilising over 10,000
students. In addition to indirect engagement of students through teachers, the FRONTIERS project
also directly engagement students through virtual visits and masterclasses. The total number of
students engaged in this in-person and online events was 1,505. teachers, it can be stated that the project also achieved its stated aim of mobilising over 10,000
students. In addition to indirect engagement of students through teachers, the FRONTIERS project
also directly engagement students through virtual visits and masterclasses. The total number of
students engaged in this in-person and online events was 1,505. Mobilisation Target The student mobilisation target for the FRONTIERS project was 10,000 within the framework of the
proposed activities. The stated mobilisation target of 10,000 students was based on the given ratio
that the mobilisation of 1 teacher would deliver the mobilisation of 10 students. This equivalence is
arrived at as teachers employ resources and training with students in the classroom, at a minimum
of 10:1. As the FRONTIERS project achieved its goal of mobilising over the stated goal of 1,000 12 12 12 In addition to the FRONTIERS Demonstrators as teaching and learning resource, the project partners
facilitated virtual visits to large-scale research facilities to engage students directly. The evaluation
results of these virtual visits delivered with the supporting Demonstrator resources, showed an
increase in student confidence with subject of Nobel prize winning physics. Analysis of findings
following virtual visits to the Virgo gravitational wave detector found students displayed a 22%
increase in familiarity with vocabulary items related to Gravitational Wave production and detection
principles as well as a 15.7% improved familiarity with items related to technical aspects of
measurement. Student Skills Development Participant 4 of the focus group welcomed this way of engaging
students with frontier physics, stating that, “Physics is a nice subject but the blackboard is not
enough to give the students the pleasure of intellectual discoveries.” Participant 4 felt that the
“practical aspect of FRONTIERS resources will instil in students the ability to ask questions and to
explore.” Participant 5 of the same focus group, had tried introducing frontier physics in a theoretical way, but
since participating in the International e-School, they had started “to introduce the topics in a
practical way with the animations and videos that are part of the demonstrators”. As a result, she
and her students feel they are “working like a scientist.” The blend of Nobel prize winning physics with inquiry-based pedagogical approach that speaks to
the professional needs of teacher was highlighted for praise also. The development of the
Demonstrators fulfilling both requirements meant “students will be using scientific language and
interpreting real graphs, using graphs in real life.” The inquiry-based learning approach to delivering
content was identified as encouraging students to employ “critical thinking, problem solving,
justifying reasoning” through “practical investigation.” 13 13 Demonstrator Internal Selection and Evaluation The first two phases of evaluation of the FRONTIERS project demonstrators were internal selection
criteria. These criteria were developed to ensure project outputs would be effective in meeting key
performance indicators and ensuring quality of deliverables. The internal evaluation and selection
criteria of Quality Criteria and Pedagogical Framework are detailed below, followed by a summary of
each of the final demonstrators selected. Institutions/Schools To assess and estimate the impact the project had at school and institution level, one of the
identified measures was the extent to which school authorities encouraged and facilitated the
participation of staff in the training provided by the FRONTIERS project. During the 15 initial teacher training events hosted by partners, 303 science teachers attended. In
addition, 277 teachers from an international audience attended the International e-School online
training events. Each of these teachers was facilitated and encouraged to attend these training
events as they were released by their schools and institutions to attend. The FRONTIERS Project also facilitated the cross-sectional cooperative community building that say
schools, universities and large-scale research institutions work collaboratively to promote Nobel
prize winning physics. During the project lifetime, 15 virtual visits were undertaken to the four large-
scale research facilities. Each of these visits was an opportunity for school communities to
experience the frontier of modern physics that teachers and students rarely are given. The virtual visits were also valuable opportunities for the large-scale research facilities to develop
their out-reach programme networks, showcasing the important work being undertaken and
encouraging the next generation of research scientists. The value of the virtual visits to research
facilities is evident from the number of visits facilitated by the four locations; Virgo, CERN, Pierre
Auger Observatory and the Faulkes Telescope. The impact of the FRONTIERS Project on schools and institutions will be further built upon through
the e-twinning opportunities supported through the Online Community Support. 14 14 Demonstrators The project delivered 21 demonstrators following multiple phases of internal selection and
evaluation criteria. Internal selection and evaluation by partners included best practice quality
criteria and compatibility with the adopted pedagogical framework. A breakdown of the selection
and evaluation criteria is included below, followed by scores for each selected demonstrator with a
short description of content and activities. Project Evaluation Each of the relevant elements project outputs were evaluated in turn. Firstly, the FRONTIERS project
demonstrators were evaluated, followed by the initial teacher training events. Next the International
e-Schools were addressed, finishing with an evaluation of the Virtual Visits facilitated by the project
partners. Best Practice Quality Criteria Possible demonstrators identified by partners were first evaluated using to the quality criteria
outlined below to ensure that only proven best practice resources would be included. As a result,
the outreach practices selected by the FRONTIERS consortium have not only been identified through
bibliographic research but also through the project partners’ hands-on experience. 15
Quality
Criteria
Appropriate
for 12-18 year
olds
Implemented
at least 3
times
Engaged at
least 100
students
Engaged at
least 10
teachers
Publicly
referenced
and visible
Utilises
frontier
science
Online tools
and
interactive
material
Adaptable to
Demonstrator Appropriate
for 12-18 year
olds Implemented
at least 3
times Implemented
at least 3
times Adaptable to
Demonstrator Online tools
and
interactive
material Publicly
referenced
and visible 15 To select the best outreach practices out of the 32 initially proposed by the project consortium, the
criteria were applied using a Google form. A link to the Google form can be found here, as well as
the feedback provided by partners. Of the initial 32 outreach practices that were evaluated using the quality criteria, 25 were proposed
as meeting the FRONTIERS best outreach practice criteria. The remainder of the outreach practices
were not chosen for inclusion, with many being quite new having low levels of impact so far. Those best-practice demonstrators that successfully passed the first evaluation criteria were then
scored according to the pedagogical approach developed and adopted by the project. These
evaluation criteria prioritised those demonstrators that adopted an inquiry-based approach to
learning. Scoring criteria under the above headings were developed for partners to evaluate each
demonstrator. Each of the demonstrators was marked against the above criteria. Other elements of the demonstrators were evaluated to enhance project impact and achievement of
stated aims. To integrate the student skills development and promotion of a positive attitude
towards STEM, the following areas were included at the initial evaluation stage. ● Learning and Innovation skills include communication, collaboration, problem solving and
thinking outside the box. ● Learning and Innovation skills include communication, collaboration, problem solving an ● Literacy skills involves the use of technology, ability to read statistics and graphs, access,
evaluate and create media. ● Life skills is a term used to describe the ability to set goals, plan use of resources, respond to
feedback, take responsibility for collecting, organizing and analysing data. Pedagogical Framework The developers of the National Science Education Standards (National Research Council, 1996)
promoted the enactment of a science curriculum that embraced an inquiry-oriented approach to
science teaching. They put forward the following essential features of inquiry-based teaching and
learning: ●
Learners are engaged by scientifically oriented questions. ●
Learners are engaged by scientifically oriented questions. ●
Learners are engaged by scientifically oriented questions. ●
Learners give priority to evidence, which allows them to develop and evaluate explanations
that address scientifically oriented questions. ●
Learners formulate explanations from evidence to address scientifically oriented questions
●
Learners evaluate their explanations in light of alternative explanations, particularly those
reflecting scientific understanding. ●
Learners communicate and justify their proposed explanations. ●
Learners communicate and justify their proposed explanations. The continuum of inquiry-based learning in science education proposed by Banchi and Bell (2008)
includes 4 types of inquiry; (i). Limited inquiry which is sometimes referred to as confirmation
inquiry, (ii). structured inquiry, (iii). guided inquiry and (iv). open inquiry. The form of scientific
inquiry embraced is determined by the level of teacher guidance. Meyer, Meyer et al. (2013) note
that teachers can have strong conceptions of what inquiry requires, and still struggle to form
instructional plans. Therefore, as suggested by Streich and Mayer (2020) guidance is pivotal in
maximising the benefits of inquiry-based learning. The Frontiers demonstrators were developed using 5 defined stages of inquiry. The Frontiers demonstrators were developed using 5 defined stages of inquiry. 1. Orienting and asking questions
2. Hypothesis generation and design
3. Planning and investigation
4. Analysis and interpretation
5. Conclusion and evaluation
A video was produced by DCU to
support the inquiry process. The video
can be accessed at
https://youtu.be/aMjmowIBrOg 16 16 Final Demonstrators The final demonstrators selected for inclusion by the FRONTIERS project are listed below, organised
according to categories of frontier physics. Following the table, a one-page description of each
demonstrator is provided, with the Pedagogical Framework score and Student Skill Development
score. It should be noted that some demonstrators facilitating a
virtual tour score relatively low on the pedagogical
framework evaluation criteria. Although not aligning with
the pedagogical framework adopted by the consortium,
these demonstrators provide invaluable opportunity for
students to witness and engage with the work of scientist in
real world settings. Each of the FRONTIERS Demonstrators is hosted on the Open Schools for Open Societies platform,
reflecting the pedagogical framework adopted by the project and making them available to the
wider community of teachers. This platform is explored further as part of the project Online
Community Support. 17 17 Non-accelerator physics
1. Cloud Chamber (EA)
2. Study cosmic rays using data from school detectors (IASA)
Gravitational Waves
3. Gravitational Wave Noise Hunting (EA)
4. Virtual Visit to the European Gravitational Observatory (EGO)
5. “Finding black-holes in a chirp”: how to understand the first gravitational-wave detection (PCCP)
6. EGO Control (Class)room (EGO)
7. Earthquake Interferometer (EGO)
Special Relativity
8. Mass-Energy Equivalence (EA)
9. Relativistic muon time dilation (EA)
Astronomy
10. Discovering Alien Worlds - The discovery of an exoplanet (NUCLIO)
11. Black Holes in My School (NUCLIO)
12. Does the Sun rotate? (NUCLIO)
13. Exploring the Sun - Solar differential rotation (NUCLIO)
Cosmology
14. “Measuring the recess velocity of distant galaxies”: calculating the age of the Universe (PCCP)
Optical Physics
15. From the Michelson-Morley experiment to the gravitational-wave detection - Discovering and building
a Michelson interferometer (PCCP)
High Energy Physics
16. The ALICE experiment at CERN (EA)
17. Search for the Z and Higgs bosons (IASA)
18. How to accelerate particles (IASA)
19. Study data from the Large Hadron Collider (IASA)
Basic Physics
20. The magnetic field and its applications (IASA)
21. The Pendulum (PCCP) 18 18 Learning outcomes: ●
To introduce students to the concept of particle detection. ●
To have students build a simple cloud chamber detector. To teach students that invisible ionizing radiation can leave visible “footprints” and what these can be. To initiate a discussion about cosmic rays. Overview This demonstrator introduces students to cloud chambers,
the first experimental apparatus that enabled scientists to
visualize elementary particles coming from the cosmos. Students are introduced to cosmic rays, they learn about the
principles of operation of a diffusion cloud chamber, and with
the help of their teacher they construct a simple diffusion
cloud chamber in order to observe cosmic ray particle tracks
in it. OSOS Link Demonstrator 1 Concepts introduced:
● Cosmic Rays:
● Muons
● Particle Detectors Concepts introduced:
● Cosmic Rays:
● Muons
● Particle Detectors Key activities: 1. Construction of a diffusion cloud chamber with simple materials. 2. Observation of particle tracks due to cosmic ray interactions with the cloud chamber. Skills Development:
7/10 19 19 Overview This scenario explains what cosmic rays are and how
they interact with the earth’s atmosphere. It talks
about the different origins of cosmic rays in respect to
their energy, what they are made of and how those
particles end up producing others inside the
atmosphere. It explains how time dilation works
according to the special theory of relativity and how
that helps muons reach the earth’s surface. Then it
explains how the students can use real data from cosmic ray detectors installed at schools all over the
world make their own plots of muon lifetime. Link OSOS Demonstrator 2
Metadata:
Age: 14-17
Duration: 4 hours
Equipment: PC with internet connection
Concepts introduced:
●
Relativity
●
Particle collisions and interactions
●
Particle lifetime
Learning outcomes:
1. Students learn about cosmic rays, what they are and where they come from
2. They learn about how they interact with the atmosphere
3. They learn how to use data and perform a simple analysis on them
Prior knowledge:
●
Elementary particles
●
Basic electromagnetism
●
Basic astronomy:
solar system, galaxies etc. Learning intentions:
●
Understand what cosmic rays are and where they come from
●
Understand how relativity affects particle lifetime
●
Use existing data to study muon lifetime
Key activities:
1. Videos and information to engage students
2. QuarkNET data and analysis tool for muon lifetime study
3. Student report and discussion
Pedagogical Framework Score:
16/18
Skills Development:
9/10
Learning Skills: 4/4
Literacy Skills: 2/3
Life Skills: 3/3 Link OSOS Demonstrator 2 20 20 Overview The discovery of Gravitational Waves provided
humankind with a new window to the universe, allowing
us to probe extreme cosmic phenomena and testing
nature at its limits. This scenario provides a step-by-step
introduction to gravitational waves and their detection,
introduces the concept of detector sensitivity and
through a game-based approach, introduces students to
the different noise parameters that affect detector
performance. us to probe extreme cosmic phenomena and testing
nature at its limits. This scenario provides a step-by-step
introduction to gravitational waves and their detection,
introduces the concept of detector sensitivity and
through a game-based approach, introduces students to
the different noise parameters that affect detector
performance. OSOS Link Demonstrator 3
Metadata:
Age: 16-18
Duration: 4 hours
Equipment: PC with internet connection
Concepts introduced:
●
Gravitational wave
●
Interferometer
●
Sources of noise
●
sensitivity
●
reach
Learning outcomes:
1. To help students understand the meaning of noise. 2. To guide students to identify sources of noise for gravitational wave detectors. 3. To help students correlate the reach of a detector with its noise level. 4. To show how to compromise budget with performance. 5. To present how to estimate the most important sources of noise and make data driven decisions. Prior knowledge:
●
Waves: Definition and
properties
●
Newton’s law of gravitation
●
Wave interference
●
Signal and noise
●
Orders of magnitude
Learning intentions:
●
describe roughly what gravitational waves are. ●
explain that detector noise deters us from exploring farther regions in the universe using
gravitational waves as messengers. ●
compare the significance of different sources of noise in a gravitational wave detector. ●
explain that even though the effects of gravitational waves on earth are minuscule, these derive
from very high energy processes. Key activities:
1. Building a virtual gravitational wave detector
2. Identifying sources of noise and taking corrective actions to reduce it
3. Searching for glitches in a gravitational wave detector
Pedagogical Framework Score:
18/18
Skills Development:
10/10
Learning Skills: 4/4
Literacy Skills: 3/3
Life Skills: 3/3 introduction to gravitational waves and their detection,
introduces the concept of detector sensitivity and
through a game-based approach, introduces students to
the different noise parameters that affect detector
performance. OSOS Link Demonstrator 3
Metadata:
Age: 16-18
Duration: 4 hours
Equipment: PC with internet connection
Concepts introduced:
●
Gravitational wave
●
Interferometer
●
Sources of noise
●
sensitivity
●
reach
Learning outcomes:
1. Overview Virgo, based in Cascina, Pisa, Italy, is one of the three
experiments in the world that is able to detect
gravitational waves. Several thousands of high-school
and university students visit the site every year since
2005. The demonstrator provides an interactive web-
based environment for making a virtual visit of Virgo
main experimental building. Students would need to
understand the optical scheme of the detector in
order to orientate themselves inside the labyrinth of
vacuum pipes. Supplementary materials about the discovery of gravitational waves, the birth of multi-messenger
astronomy, the operating principle of the detector and its noises is also given. OSOS Link Demonstrator 4
Metadata:
Age: > 16
Duration: 3 hours
Equipment: PC with internet connection
Concepts introduced:
● Space-time ● Gravitational Wave
● Black-hole ● Interferometer
● Laser ● Seismic Noise
Learning outcomes:
1. Teach students about the discovery of gravitational waves and its importance to improve our
understanding of the universe. 2. Give an idea of the experimental challenges of gravitational wave detection. 3. Introduce basic concepts of General Relativity and Astrophysics. Prior knowledge:
● Newtonian Mechanics
Learning intentions:
●
Have an idea of what are gravitational waves
●
Have an idea of what is a laser interferometer and how it works
Key activities:
1. Slides with pictures and videos to engage
2. Understand Virgo optical scheme
3. Treasure hunting inside the interferometer experimental building
4. Impressions and discussion on exploration of the detector
5. Final report and discussion
Pedagogical Framework Score:
5/18
Skills Development:
8/10
Learning Skills: 4/4
Literacy Skills: 3/3
Life Skills: 1/3 OSOS Link Demonstrator 4
Metadata:
Age: > 16
Duration: 3 hours
Equipment: PC with internet connection
Concepts introduced:
● Space-time ● Gravitational Wave
● Black-hole ● Interferometer
● Laser ● Seismic Noise OSOS Link Demonstrator 4 Overview To help students understand the meaning of noise. 2. To guide students to identify sources of noise for gravitational wave detectors. 3. To help students correlate the reach of a detector with its noise level. 4. To show how to compromise budget with performance. 5. To present how to estimate the most important sources of noise and make data driven decisions. Prior knowledge:
●
Waves: Definition and
properties
●
Newton’s law of gravitation
●
Wave interference
●
Signal and noise
●
Orders of magnitude
Learning intentions:
●
describe roughly what gravitational waves are. ●
explain that detector noise deters us from exploring farther regions in the universe using
gravitational waves as messengers. ●
compare the significance of different sources of noise in a gravitational wave detector. ●
explain that even though the effects of gravitational waves on earth are minuscule, these derive
from very high energy processes. Key activities:
1. Building a virtual gravitational wave detector
2. Identifying sources of noise and taking corrective actions to reduce it
3. Searching for glitches in a gravitational wave detector
Pedagogical Framework Score:
18/18
Skills Development:
10/10
Learning Skills: 4/4
Literacy Skills: 3/3
Life Skills: 3/3 21 21 Learning outcomes: g
1. Teach students about the discovery of gravitational waves and its importance to improve our
understanding of the universe. 22 22 Demonstrator 5: Finding black holes in a chirp
How to understand the first gravitational-wave detection Demonstrator 5: Finding black holes in a chirp
How to understand the first gravitational-wave detection Overview Overview
This demonstrator introduces the concept of
gravitational waves, their possible sources, and
the way they have been detected. Students are
introduced to the basics of data analysis using
the real signal of the first gravitational wave ever
detected, produced by the motion of two
inspiralling masses. Students will learn how to determine the masses
and the radius of the binary system, to identify
the two objects as black holes, and what are the fundamental properties and parameters of a black
hole. This demonstrator introduces the concept of
gravitational waves, their possible sources, and
the way they have been detected. Students are
introduced to the basics of data analysis using
the real signal of the first gravitational wave ever
detected, produced by the motion of two
inspiralling masses. p
g
Students will learn how to determine the masses
and the radius of the binary system, to identify
the two objects as black holes, and what are the fundamental properties and parameters of a black
hole OSOS Link Demonstrator 5
Metadata:
Age: >17
Duration: 3 hours
Equipment: PC with internet connection
Concepts introduced:
●
Basics of general relativity
●
Gravitational-wave emission
●
Binary black holes systems
●
Schwarzschild radius
●
Chirp mass
Learning outcomes:
1. Learn the basics of general relativity, black holes, and gravitational-wave emission and detection. 2. understand the methodology and reproduce the analyses behind the discovery of gravitational
waves. 3. Introduce basic concepts of astrophysics and gravitational-wave astronomy. Prior knowledge:
●
Algebra
●
Basic concept of calculus
●
Wave properties
●
Knowledge of Newton and
Kepler laws
Learning intentions:
●
Define a gravitational wave and interpret its signal
●
Explain the gravitational waves observation method
●
Calculate the Schwarzschild radius of an astrophysical object. ●
Explain the merger of a binary system and draw a scheme of the gravitational signal emitted. Key activities:
1. Videos to engage
2. Data analysis and explanation
3. Optional: python scripts for data analysis
4. Final report and discussion
Pedagogical Framework Score:
16/18
Skills Development:
6/10
Learning Skills: 2/4
Literacy Skills: 2/3
Life Skills: 2/3 OSOS Link Demonstrator 5
Metadata:
Age: >17
Duration: 3 hours
Equipment: PC with internet connection
Concepts introduced:
●
Basics of general relativity
●
Gravitational-wave emission
●
Binary black holes systems
●
Schwarzschild radius
●
Chirp mass
Learning outcomes:
1. Learn the basics of general relativity, black holes, and gravitational-wave emission and detection. 2. Overview Analysis of recent data in respect to the environment (Wind speed, Sea activit)
5. Final report and discussion
Pedagogical Framework Score:
13/18
Skills Development:
8/10
Learnin Demonstrator 6: Ego Control (Class)room
Overview
All big experiments like LHC or Virgo have a control
room. It is there that all the magic happens! There all
the troubles, successes, joy and despair of the
experimental physicist life are concentrated. In this
room tens of people workday and night together to
improve the sensitivity of the detectors. The
demonstrator allows the teacher to have in your class
many of the screens present in the Virgo control room. This allows for several studies aimed at understanding the
environment. This activity is very close to the work done da
Supplementary materials about the discovery of gravitatio
astronomy, the operating principle of the detector and its
OSOS Link Demonstrator 6
Metadata:
Age: >16
Duration: 3 hours
Equipment: PC with internet connection
Conce
Gravit
Obser
Seism
Learning outcomes:
1. Give an idea of the experimental challenges of gra
2. Give an idea of the work that physicists do in the V
3. Let students understand the strong interaction be
Prior knowledge:
● Newtonian Mechanics
Learning intentions:
●
What are gravitational waves? ●
What is a laser interferometer and how it works
●
What are the noises that disturb the detector
Key activities:
1. Slides with pictures and videos to engage
2. Configure the PC or RPi to show Virgo real time data
3. Familiarize with the Control Room tools (DMS, VIM, …
4. Analysis of recent data in respect to the environment
5. Final report and discussion
Pedagogical Framework Score:
13/18
Skills D Overview All big experiments like LHC or Virgo have a control
room. It is there that all the magic happens! There all
the troubles, successes, joy and despair of the
experimental physicist life are concentrated. In this
room tens of people workday and night together to
improve the sensitivity of the detectors. The
demonstrator allows the teacher to have in your class
many of the screens present in the Virgo control room. This allows for several studies aimed at understanding the relationship between the detector and the
environment. This activity is very close to the work done day by day by the physicists working on site. Supplementary materials about the discovery of gravitational waves, the birth of multi-messenger
astronomy, the operating principle of the detector and its noises is also given. OSOS Link Demonstrator 6
Metadata:
Age: >16
Duration: 3 hours
Equipment: PC with internet connection
Concepts introduced:
Gravitational Wave Interferometer
Observing range (Parsec) Fourier transform
Seismic Noise
Learning outcomes:
1. Give an idea of the experimental challenges of gravitational wave detection. 2. Give an idea of the work that physicists do in the Virgo control room. 3. Let students understand the strong interaction between the environment and the detector. Prior knowledge:
● Newtonian Mechanics
Learning intentions:
●
What are gravitational waves? ●
What is a laser interferometer and how it works
●
What are the noises that disturb the detector
Key activities:
1. Slides with pictures and videos to engage
2. Configure the PC or RPi to show Virgo real time data
3. Familiarize with the Control Room tools (DMS, VIM, …)
4. Analysis of recent data in respect to the environment (Wind speed, Sea activit)
5. Final report and discussion
Pedagogical Framework Score:
13/18
Skills Development:
8/10
Learning Skills: 4/4
Literacy Skills: 3/3
Life Skills: 1/3 Overview understand the methodology and reproduce the analyses behind the discovery of gravitational
waves. 3. Introduce basic concepts of astrophysics and gravitational-wave astronomy. Prior knowledge:
●
Algebra
●
Basic concept of calculus
●
Wave properties
●
Knowledge of Newton and
Kepler laws
Learning intentions:
●
Define a gravitational wave and interpret its signal
●
Explain the gravitational waves observation method
●
Calculate the Schwarzschild radius of an astrophysical object. ●
Explain the merger of a binary system and draw a scheme of the gravitational signal emitted. Key activities:
1. Videos to engage
2. Data analysis and explanation
3. Optional: python scripts for data analysis
4. Final report and discussion
Pedagogical Framework Score:
16/18
Skills Development:
6/10
Learning Skills: 2/4
Literacy Skills: 2/3
Life Skills: 2/3 Skills Development:
6/10 23 23 Demonstrator 6: Ego Control (Class)room
Overview
All big experiments like LHC or Virgo have a control
room. It is there that all the magic happens! There all
the troubles, successes, joy and despair of the
experimental physicist life are concentrated. In this
room tens of people workday and night together to
improve the sensitivity of the detectors. The
demonstrator allows the teacher to have in your class
many of the screens present in the Virgo control room. This allows for several studies aimed at understanding the relationship between the de
environment. This activity is very close to the work done day by day by the physicists w
Supplementary materials about the discovery of gravitational waves, the birth of multi
astronomy, the operating principle of the detector and its noises is also given. OSOS Link Demonstrator 6
Metadata:
Age: >16
Duration: 3 hours
Equipment: PC with internet connection
Concepts introduced:
Gravitational Wave Inte
Observing range (Parsec) Four
Seismic Noise
Learning outcomes:
1. Give an idea of the experimental challenges of gravitational wave detection. 2. Give an idea of the work that physicists do in the Virgo control room. 3. Let students understand the strong interaction between the environment and
Prior knowledge:
● Newtonian Mechanics
Learning intentions:
●
What are gravitational waves? ●
What is a laser interferometer and how it works
●
What are the noises that disturb the detector
Key activities:
1. Slides with pictures and videos to engage
2. Configure the PC or RPi to show Virgo real time data
3. Familiarize with the Control Room tools (DMS, VIM, …)
4. OSOS Link Demonstrator 6 OSOS Link Demonstrator 6
Metadata:
Age: >16
Duration: 3 hours
Equipment: PC with internet connection
Concepts introduced:
Gravitational Wave Interferometer
Observing range (Parsec) Fourier transform
Seismic Noise
Learning outcomes:
1. Give an idea of the experimental challenges of gravitational wave detection. 2. Give an idea of the work that physicists do in the Virgo control room. 3. Let students understand the strong interaction between the environment and the detector. Prior knowledge:
● Newtonian Mechanics
Learning intentions:
●
What are gravitational waves? ●
What is a laser interferometer and how it works
●
What are the noises that disturb the detector
Key activities:
1. Slides with pictures and videos to engage
2. Configure the PC or RPi to show Virgo real time data
3. Familiarize with the Control Room tools (DMS, VIM, …)
4. Analysis of recent data in respect to the environment (Wind speed, Sea activit)
5. Final report and discussion
Pedagogical Framework Score:
13/18
Skills Development:
8/10
Learning Skills: 4/4
Literacy Skills: 3/3
Life Skills: 1/3 24 24 Demonstrator 7: Earthquake Interferometer
Overview
Using the data shown in Control (Class)room
demonstrator several studies aimed at
understanding the relationship between the
detector and the environment can be made. The
correlation between the sensitivity of the detector
and the arrival of strong remote earthquakes can
be studied. This allows the students to understand
both the nature and propagation speed of seismic waves and at the same time to experience directly
how a gravitational interferometer work. Moreover, this activity is very close to the work done day by
day by the physicists working on site. Overview Overview
Using the data shown in Control (Class)room
demonstrator several studies aimed at
understanding the relationship between the
detector and the environment can be made. The
correlation between the sensitivity of the detector
and the arrival of strong remote earthquakes can
be studied. This allows the students to understand
both the nature and propagation speed of seismic waves and at the same time to experience directly
how a gravitational interferometer work. Moreover, this activity is very close to the work done day by
day by the physicists working on site. OSOS Link Demonstrator 7
Metadata:
Age: >16
Duration: 3 hours
Equipment: PC with internet connection
Concepts introduced:
Gravitational Wave Interferometer
Logarithmic scales Fourier transform
Harmonic oscillator Seismic Noise
Seismic waves & Earthquakes
Learning outcomes:
1. Give an idea of the experimental challenges of gravitational wave detection. 2. Give an idea of the work that physicists do in the Virgo control room. 3. Let students understand the strong interaction between the environment and the detector. Prior knowledge:
● Newtonian Mechanics
Learning intentions:
●
What are gravitational waves
●
What is a laser interferometer and how it works
●
How earthquakes affect GW interferometers
Key activities:
1. Slides with pictures and videos to engage
2. Configure the PC or RPi to show Virgo real time data
3. Familiarize with the Control Room tools (DMS, VIM, …)
4. Analysis of the effect of recent earthquakes on the observing range of the detector
5. Final report and discussion
Pedagogical Framework Score:
15/18
Skills Development:
8/10
Learning Skills: 4/4
Literacy Skills: 3/3
Life Skills: 1/3 OSOS Link Demonstrator 7
Metadata:
Age: >16
Duration: 3 hours
Equipment: PC with internet connection
Concepts introduced:
Gravitational Wave Interferometer
Logarithmic scales Fourier transform
Harmonic oscillator Seismic Noise
Seismic waves & Earthquakes
Learning outcomes:
1. Give an idea of the experimental challenges of gravitational wave detection. 2. Give an idea of the work that physicists do in the Virgo control room. 3. Let students understand the strong interaction between the environment and the detector. Prior knowledge:
● Newtonian Mechanics
Learning intentions:
●
What are gravitational waves
●
What is a laser interferometer and how it works
●
How earthquakes affect GW interferometers
Key activities:
1. Slides with pictures and videos to engage
2. Configure the PC or RPi to show Virgo real time data
3. Familiarize with the Control Room tools (DMS, VIM, …)
4. Overview Analysis of the effect of recent earthquakes on the observing range of the detector
5. Final report and discussion
Pedagogical Framework Score:
15/18
Skills Development:
8/10
Learning Skills: 4/4
Literacy Skills: 3/3
Life Skills: 1/3 OSOS Link Demonstrator 7 Concepts introduced:
Gravitational Wave Interferometer
Logarithmic scales Fourier transform
Harmonic oscillator Seismic Noise
Seismic waves & Earthquakes 25 25 Overview This demonstrator introduces the most famous equation
in Physics, E = mc2 to students. Students form a research
question about the meaning of mass-energy
equivalence, and they explore the equation and its
applications. Students examine the units and
calculations required to apply Einstein’s equation to real
life. Students gain experience of completing calculations
and compare their results to the experimentally
collected data. Students understand that the law of
conservation of energy and the law of conservation of mass don’t apply individually but together. They watch videos which explain this effect and its applications to real world phenomena. OSOS Link Demonstrator 8
Metadata:
Age: 15-17
Duration: 3 hours
Equipment: PC with internet connection
Concepts introduced:
Mass-Energy Nuclear power
Radioactivity The electronvolt unit
The atomic mass unit
Learning outcomes:
1. Introduce students to the most famous equation in the history of Physics. 2. Introduction to the basic principles behind nuclear fission using mass-energy equivalence. 3. Demonstrate that energy and mass are not conserved separately in nature. Prior knowledge:
● Concept of mass
● Concept of energy
● Chemical formula
● Linear equations
Learning intentions:
●
Identify the significance of Einstein’s equation
●
List examples of processes where energy is created according to E=mc2
●
Change the units of energy between eV and J
●
Change the mass of a particle from amu to kg
●
Calculate loss/gain in mass from a chemical reaction and calculate the energy created/required
●
Explain the relationship between mass and energy
Key activities:
1. Matching activity of key words to images
2. Converting units’ calculations
3. Develop hypothesis and test using calculations
4. Summary presentation
Pedagogical Framework Score:
17/18
Skills Development:
6/10
Learning Skills: 3/4
Literacy Skills: 2/3
Life Skills: 1/3 26 26 Overview Overview
This demonstrator introduces students to the phenomenon of
relativistic time dilation, one of the most striking consequences of
Einstein’s Special Theory of Relativity. Utilizing a virtual simulation,
students will investigate the rate of decay in flight of muons produced
in elementary particle collisions in the atmosphere and will be guided
to explain their findings using Einstein’s theory of Special Relativity and
the phenomenon of time dilation. This scenario is a follow-up from the
educational scenario on cosmic muon decay. OSOS Link Demonstrator 9
Metadata:
Age: 16-18
Duration: 3 hours
Equipment: PC with internet connection
Concepts introduced:
Muon Decay in flight
Relativistic Time
Dilation
Flux
Learning outcomes:
●
To introduce students to the concept of radioactive decay. ●
To introduce students to cosmic ray muons. ●
To introduce students to relativistic time dilation. ●
To initiate a discussion about cosmic rays. Prior knowledge:
Exponential decay
Radioactivity
Properties of muons,
Exponential functions
The “eV” unit of energy and its
multiples. Learning intentions:
●
Discuss the properties of muons. ●
Discuss the phenomenon of radioactive decay. ●
Explain the phenomenon of relativistic time dilation in simple terms. ●
Plot data in excel and fit them with an exponential function. ●
Explain the concept of flux and be able to calculate it for given parameters
Key activities:
1. Performing simple kinematics calculations
2. Performing a simple counting experiment for different altitudes. 3. Collecting data, plotting them and fitting them
4. Develop hypothesis and test using calculations
5. Summary presentation
Pedagogical Framework Score:
17/18
Skills Development:
7/10
Learning Skills: 3/4
Literacy Skills: 3/3
Life Skills: 1/3 This demonstrator introduces students to the phenomenon of
relativistic time dilation, one of the most striking consequences of
Einstein’s Special Theory of Relativity. Utilizing a virtual simulation,
students will investigate the rate of decay in flight of muons produced
in elementary particle collisions in the atmosphere and will be guided
to explain their findings using Einstein’s theory of Special Relativity and
the phenomenon of time dilation. This scenario is a follow-up from the
educational scenario on cosmic muon decay. the phenomenon of time dilation. This scenario is a follow-up from the
educational scenario on cosmic muon decay. OSOS Link Demonstrator 9
Metadata:
Age: 16-18
Duration: 3 hours
Equipment: PC with internet connection
Concepts introduced:
Muon Decay in flight
Relativistic Time
Dilation
Flux
Learning outcomes:
●
To introduce students to the concept of radioactive decay. Overview ●
To introduce students to cosmic ray muons. ●
To introduce students to relativistic time dilation. ●
To initiate a discussion about cosmic rays. Prior knowledge:
Exponential decay
Radioactivity
Properties of muons,
Exponential functions
The “eV” unit of energy and its
multiples. Learning intentions:
●
Discuss the properties of muons. ●
Discuss the phenomenon of radioactive decay. ●
Explain the phenomenon of relativistic time dilation in simple terms. ●
Plot data in excel and fit them with an exponential function. ●
Explain the concept of flux and be able to calculate it for given parameters
Key activities:
1. Performing simple kinematics calculations
2. Performing a simple counting experiment for different altitudes. 3. Collecting data, plotting them and fitting them
4. Develop hypothesis and test using calculations
5. Summary presentation
Pedagogical Framework Score:
17/18
Skills Development:
7/10
Learning Skills: 3/4
Literacy Skills: 3/3
Life Skills: 1/3 The “eV” unit of energy and its
multiples. Skills Development:
7/10 27 27 Overview This demonstrator introduces the concept of an
exoplanet and how they have been discovered in our
universe. Students are introduced to the transit method
of exoplanet detection. Real images will be used to look
for changes in the starlight that might result due to the
motion of an orbiting exoplanet. Students will be given
real light images and learn how to use a specific image
software to perform photometry and to analyse the
graphs. The analysis of the light curve will allow students to estimate the rotational period of the
exoplanet and its diameter. Students can present their work to the class and discuss how they
compare with the most accurate results that astronomers have. OSOS Link Demonstrator 10
Metadata:
Age: 14-18
Duration: 3 hours
Equipment: PC with internet connection, light
sensor and basic lab equipment
Concepts introduced:
●
Exoplanets
●
Luminosity
●
Graphing
Learning outcomes:
1. Teach students about the discovery of exoplanets
2. Allow students to understand the science and methodology behind the discovery of alien worlds. 3. Introduce basic concepts of Astronomy and Image Processing
Prior knowledge:
● Basic Astronomy knowledge of a star and planet system
Learning intentions:
●
Define an exoplanet
●
Explain the transit method
●
Describe a method to collect luminosity data
●
Examine images using the salsa J software
●
Form conclusions about a planet from a light graph
Key activities:
1. Videos to engage - possible edpuzzle
2. Explaining the light curve
3. Practical formation of a light curve
4. Salsa J - examination of 3 stars
5. Analysis and explanation
6. Final report
Pedagogical Framework Score:
16/18
Skills Development:
9/10
Learning Skills: 4/4
Literacy Skills: 2/3
Life Skills:3/3 OSOS Link Demonstrator 10 28 28 Overview This demonstrator introduces the concept of black holes
and how difficult it is to find them. Students are
introduced to a method of detecting black holes in
eclipsing binary systems – systems formed by a visible
star and an eclipsing companion. Real images will be used
to look for changes in the starlight that might result due
to the presence of a companion. Students will learn how
to use a specific image software to perform photometry
on the images and will create a graph of brightness
variation. The analysis of the light curve will allow
students to estimate the orbital period of the companion and, given a relation between several
parameters of the known star and the companion’s mass, they will estimate the minimum mass of
the unknown companion and decide if it is a strong candidate to be a black hole. Students can
present their work to the class and discuss how they compare with the most accurate results that
astronomers have. OSOS Link Demonstrator 11
Metadata:
Age: 14-18
Duration: 3 hours
Equipment: PC with internet connection
Concepts introduced:
Black holes
Luminosity
Photometry
Graph analysis
Learning outcomes:
1. Teach students about black holes and how to detect them. 2. Allow students to understand the science and methodology behind the detection of black holes. 3. Introduce basic concepts of Astronomy and Image Processing
Prior knowledge:
Basic Astronomy knowledge
Excel (charts)
Kinematic concepts:
radial velocity, orbital period, Keppler’s Law
Learning intentions:
●
Define a black hole
●
Explain an eclipsing binary system
●
Describe a method to collect luminosity data
●
Examine images using the salsa J software
Key activities:
1. Videos to engage - possible edpuzzle
2. Explaining the light curve
3. Practical formation of a light curve
4. Salsa J - examination of 3 stars
5. Analysis and explanation
6. Final Report
Pedagogical Framework Score:
17/18
Skills Development:
9/10
Learning Skills: 4/4
Literacy Skills: 2/3
Life Skills: 3/3 29 29 Demonstrator 13: Exploring the Sun - Solar differential rotation Demonstrator 13: Exploring the Sun - Solar differential rotation Demonstrator 12: Exploring the Sun - Does the Sun rotate? This demonstrator introduces the Sun as an active star with sunspots. Students will learn how to use a specific image software to explore
sets of real images to understand better sunspots (they will measure
sunspots and compare with Earth). The sets of images will lead
students to say that the Sun rotates. But they are confronted with
images of a transit of an inner planet. Students will have to find a
way to argue that sunspots are not satellites - they belong to the
Sun’s surface and therefore show that the Sun rotates. Students can
present their work to the class and discuss how Galileo Galilei solved
this dilemma. OSOS Link Demonstrator 12
Metadata:
Age: 14-17
Duration: 2 hours
Equipment: PC with internet connection
Concepts introduced:
●
Sun’s activity
●
sunspots
Learning outcomes:
1. Teach students about the Sun’s activity and its influence on Earth. 2. Allow students to understand the evidence of the Sun’s rotation movement. 3. Introduce basic concepts of Astronomy and Image Processing. 4. Recognize that sometimes the interpretation of the data analysis is not straightforward. Prior knowledge:
●
Basic Astronomy knowledge of the solar system (Sun, planets, satellites)
●
Scales
Learning intentions:
●
Describe the Sun as an active star
●
Explain what sunspots are
●
Examine images using the salsa J software
●
Form conclusions about evidences that the Sun rotates
Key activities:
1. Videos to engage
2. Salsa J - making movies and examining images
3. Analysis and explanation
4. Final report and discussion
Pedagogical Framework Score:
16/18
Skills Development:
9/10
Learning Skills: 4/4
Literacy Skills: 2/3
Life Skills: 3/3 OSOS Link Demonstrator 12 30 30 Overview Students will
present their work to the class and discuss how it compare with the
most accurate results that astronomers have. OSOS Link Demonstrator 14
Metadata:
Age: > 16
Duration: 3 hours
Equipment: PC with internet connection
Concepts introduced:
● Spectral lines analysis
● Astronomical redshift
● Universe’s expansion and a
● Cosmic Microwave Backgro
Learning outcomes:
1. Teach students about the expansion of the Universe and the basics of Hu
2. Allow students to understand the daily work of an observational astrono
3. Introduce basic concepts of observational astronomy, cosmology, and im
Prior knowledge:
● Algebra
● Wave properties
● Basics of l
Learning intentions:
●
Explain the expansion of the Universe and its observational discovery
●
Remember the Doppler Effect and the interpretation of redshift. ●
Describe a spectrum and interpret its absorption lines. ●
Explain the idea of standard candles in astronomy. ●
Interpret data evaluating signal/noise ratio. Key activities:
1. Videos to engage
2. Data taking, analysis, and interpretation
3. Final report and discussion
Pedagogical Framework Score:
Skills Development: Overview O e
e
This demonstrator introduces the Sun as an
active star and how its activity can influence
Earth. Students are questioned about its
movements and led to realize that the Sun
rotates. Real images will be used to determine
the solar rotation period. Students will learn how
to use a specific image software to track
sunspots and calculate the Sun’s rotation period. Further analysis will show that the Sun doesn’t
rotate as a rigid body: it exhibits differential rotation. Students can present their work to the class and
discuss how they compare with the most accurate results that astronomers have. OSOS Link Demonstrator 13 OSOS Link Demonstrator 13
Metadata:
Age: 14-17
Duration: 3 hours
Equipment: PC with internet connection
Concepts introduced:
● Sun’s activity
● Sunspots
Learning outcomes:
1. Teach students about the Sun’s activity and its influence on Earth. 2. Allow students to understand the evidence of the Sun’s rotation movement. 3. Introduce basic concepts of Astronomy and Image Processing. Prior knowledge:
● Basic Astronomy knowledge of the Sun as a
star
● Excel (charts)
● Kinematic concepts such as: period
Learning intentions:
●
Describe the Sun as an active star
●
Explain what sunspots are
●
Describe a method to measure the rotation period of the Sun
●
Examine images using the salsa J software
●
Form conclusions about the Sun’s differential rotation
●
Key activities:
1. Videos to engage
2. Salsa J - making movies and examining images
3. Different methods to measure the rotation period of the Sun
4. Analysis and explanation
5. Final report and discussion
Pedagogical Framework Score:
15/18
Skills Development:
9/10
Learning Skills: 4/4
Literacy Skills: 2/3
Life Skills: 3/3 31 31 Demonstrator 14: Measuring the Recess Velocity of Dista
Overview
This demonstrator introduces the concept of the expansion of the
Universe discovered by Edwin Hubble. Students will reproduce
Hubble’s measurements in order to obtain an estimate of the age of
the Universe, thanks to real sky images and data taking. They will play
the role of an observational astronomer and learn about some
fundamental properties of astronomical sources, such as the
magnitude and the luminosity distance. They will also learn about
light spectra, absorption lines, redshift, and the Doppler effect. Students will reproduce the Hubble diagram and get a measurement
of the Hubble constant and the age of the Universe. OSOS Link Demonstrator 15 OSOS Link Demonstrator 15
Metadata:
Age: > 16
Duration: 3 hours
Equipment: PC with internet connection
Concepts introduced:
● Light interference
● Gravitational-wave detection
● Power of interferometry
Learning outcomes:
1. Give students an historical view of two Nobel Prize winning discoveries using the interferometer
and its application to gravitational-wave detection. 2. Allow students to understand the methodology of experimental physics by building, calibrating,
and operating an instrument such as a Michelson interferometer. 3. Introduce basic concepts of electromagnetism and light interferometry. Prior knowledge:
● Wave properties
● Basics of classical optics
Learning intentions:
●
Define the principles of light interference
●
Explain how it can be used to measure distances
●
Test and understand the response of an instrument
Key activities:
1. Michelson interferometer manipulation
2. Videos to engage
3. Data analysis and explanation
4. Final report and discussion
Pedagogical Framework Score:
17/18
Skills Development:
10/10
Learning Skills: 4/4
Literacy Skills: 3/3
Life Skills: 3/3 Overview This demonstrator introduces the concept of waves,
interference, and the wave nature of light. Students are
introduced to the history of the Michelson
interferometer and the Michelson-Morley experiment. They will learn how the same instrument was used to
detect gravitational waves, thus confirming one of the
main predictions of Einstein’s general relativity. Students will experiment with a real small-scale
Michelson interferometer, a powerful instrument that
uses light interference to measure distances with high precision. They will learn about the basic
properties of light interference and the working principle of an interferometer. Real images from the
sites of the LIGO and Virgo instruments will be used to explain how modern-day interferometers are
used to detect gravitational waves. Students will present their work to the class and discuss their
results. Demonstrator 14: Measuring the Recess Velocity of Distant Galaxies This demonstrator introduces the concept of the expansion of the
Universe discovered by Edwin Hubble. Students will reproduce
Hubble’s measurements in order to obtain an estimate of the age of
the Universe, thanks to real sky images and data taking. They will play
the role of an observational astronomer and learn about some
fundamental properties of astronomical sources, such as the
magnitude and the luminosity distance. They will also learn about
light spectra, absorption lines, redshift, and the Doppler effect. Students will reproduce the Hubble diagram and get a measurement
of the Hubble constant and the age of the Universe. Students will
present their work to the class and discuss how it compare with the
most accurate results that astronomers have. OSOS Link Demonstrator 14
Metadata:
Age: > 16
Duration: 3 hours
Equipment: PC with internet connection
Concepts introduced:
● Spectral lines analysis ● Doppler effect
● Astronomical redshift ● Standard candles
● Universe’s expansion and age
● Cosmic Microwave Background
Learning outcomes:
1. Teach students about the expansion of the Universe and the basics of Hubble observations. 2. Allow students to understand the daily work of an observational astronomer. 3. Introduce basic concepts of observational astronomy, cosmology, and image processing. Prior knowledge:
● Algebra
● Wave properties
● Basics of light spectrum
Learning intentions:
●
Explain the expansion of the Universe and its observational discovery
●
Remember the Doppler Effect and the interpretation of redshift. ●
Describe a spectrum and interpret its absorption lines. ●
Explain the idea of standard candles in astronomy. ●
Interpret data evaluating signal/noise ratio. Key activities:
1. Videos to engage
2. Data taking, analysis, and interpretation
3. Final report and discussion
Pedagogical Framework Score:
15/18
Skills Development:
9/10
Learning Skills: 3/4
Literacy Skills: 3/3
Life Skills: 3/3 OSOS Link Demonstrator 14 32 32 Demonstrator 15: Discovering and Building a Michelson Interferometer Learning outcomes: Skills Development:
10/10 33 33 Overview The “ALICE Experiment at LHC” demonstrator, connects students
with fundamental research taking place in one of the 4 large
experiments of the LHC complex at CERN, ALICE, which
reproduces and studies the state of the Universe at a tiny
fraction of a second after the Big Bang: The Quark Gluon Plasma. Students learn about the research done at CERN and by working
in teams, they explore and analyse real scientific data and
search for the Signatures of Quark Gluon Plasma in the ALICE
detector. g
g
Students learn about the research done at CERN and by working
in teams, they explore and analyse real scientific data and
search for the Signatures of Quark Gluon Plasma in the ALICE
detector. OSOS Link Demonstrator 16
Metadata:
Age: 16 - 18
Duration: 4 hours
Equipment: PC with internet connection
Concepts introduced:
● Quark gluon plasma
● Strange particles
● Invariant mass
● Antimatter
Learning outcomes:
1. To demonstrate what Quark Gluon Plasma is. 2. To explain that through a high energy particle collision, students can study the Universe at its
infancy. 3. To employ the conservation of momentum and energy to understand data produced by subatomic
particle collisions. 4. To explain the internal structure of matter and the importance of fundamental particles such as
strange particles, which are not part of our everyday world, in understanding the structure and the
Laws of the Universe at its infancy. Prior knowledge:
● Mass – energy equivalence
● Structure of the atom
● Conservation of momentum
● Conservation of energy
● What is a histogram
Learning intentions:
●
Explain in general terms what Quark Gluon Plasma is and how we can detect it
●
Explain in general terms what strangeness is
●
Explain in general terms what is the mission of the ALICE experiment at CERN
●
Understand that particles and their anti-particles are not the same but have equal masses. Key activities:
1. Observing histograms and extracting scientific information from them. 2. Identifying particles according to their properties in a virtual detector. 3. Creating histograms. 4. Summary presentation. Pedagogical Framework Score:
16/18
Skills Development:
7/10
Learning Skills: 3/4
Literacy Skills: 3/3
Life Skills: 1/3 Learning outcomes: 34 34 Overview Overview
This exercise introduces students to CERN, the
LHC accelerator and the ATLAS experiment. It
explains the basics of the standard model of
particle physics and particle decays, more
specifically Z and Higgs bosons. It explains how
particles can be identified by their signatures and
what invariant masses and histograms are. It
gives instructions on how to use the HYPATIA
event display to analyse data from ATLAS, specifically Z and Higgs boson decays. At the end through a
series of questions and discussions directs students to present their findings. OSOS Link Demonstrator 17
Metadata:
Age: 14-17
Duration: 4 hours
Equipment: PC with internet connection
Concepts introduced:
●
Particle decays
●
Relativistic mass/invariant mass
●
Histograms
Learning outcomes:
1. Students learn about CERN, LHC and ATLAS
2. They become familiar with the standard model of particle physics and particle decays
3. They learn how to analyse data from the ATLAS experiment to look for specific particle signatures
4. They are asked to present their results and justify the outcome of their investigation
Prior knowledge:
● Fundamental forces
● Structure of matter
Learning intentions:
●
Understand the research being conducted at CERN and ATLAS
●
Understand how the LHC works
●
Learn how particle collisions can produce new particles
●
Identify particles by their signature
●
Understand and interpret the results of their investigation
Key activities:
1. Videos about CERN, LHC, ATLAS
2. Explanation about the standard model, particle collisions and decays
3. Investigation through the use of HYPATIA
4. Presentation of student investigation results
Pedagogical Framework Score:
17/18
Skills Development:
8/10
Learning Skills: 4/4
Literacy Skills: 2/3
Life Skills: 2/3 OSOS Link Demonstrator 17
Metadata:
Age: 14-17
Duration: 4 hours
Equipment: PC with internet connection
Concepts introduced:
●
Particle decays
●
Relativistic mass/invariant mass
●
Histograms
Learning outcomes:
1. Students learn about CERN, LHC and ATLAS
2. They become familiar with the standard model of particle physics and particle decays
3. They learn how to analyse data from the ATLAS experiment to look for specific particle signatures
4. Overview They are asked to present their results and justify the outcome of their investigation
Prior knowledge:
● Fundamental forces
● Structure of matter
Learning intentions:
●
Understand the research being conducted at CERN and ATLAS
●
Understand how the LHC works
●
Learn how particle collisions can produce new particles
●
Identify particles by their signature
●
Understand and interpret the results of their investigation
Key activities:
1. Videos about CERN, LHC, ATLAS
2. Explanation about the standard model, particle collisions and decays
3. Investigation through the use of HYPATIA
4. Presentation of student investigation results
Pedagogical Framework Score:
17/18
Skills Development:
8/10
Learning Skills: 4/4
Literacy Skills: 2/3
Life Skills: 2/3 OSOS Link Demonstrator 17 35 35 Overview This exercise in an introduction to particle accelerators
and high energy physics. The focus of the exercise is
the world’s most powerful accelerator, the LHC, which
is located at CERN. Students learn about how it
accelerates, bends and focuses proton beams and how
it makes sure that the protons collide inside one of the
four detectors located on its circumference. To emphasise what they’ve learned after the lesson,
the students play a game in which they have to apply
what they’ve learned in a ‘virtual accelerator’. This exercise in an introduction to particle accelerators
and high energy physics. The focus of the exercise is
the world’s most powerful accelerator, the LHC, which
is located at CERN. Students learn about how it
accelerates, bends and focuses proton beams and how
it makes sure that the protons collide inside one of the
four detectors located on its circumference. To emphasise what they’ve learned after the lesson,
the students play a game in which they have to apply
what they’ve learned in a ‘virtual accelerator’. OSOS Link Demonstrator 18
Metadata:
Age: 6-12
Duration: 2 hours
Equipment: PC with internet connection
Concepts introduced:
● Fundamental Forces
● Magnetic and Electric Fields
● Particle Accelerators
Learning outcomes:
1. To familiarise students with the basic concepts of particle accelerators
2. To introduce students to the high energy physics
3. To introduce students to CERN
Prior knowledge:
● Fundamental forces
● Atom Structure and particles
● Basic electrostatics
Learning intentions:
●
Explain the fundamental forces
●
Explain how electric fields can be used to accelerate particles
●
Explain how magnetic fields can direct particles in the LHC
●
Describe how the LHC accelerates particles
●
List practical uses of the LHC
Key activities:
1. Videos to engage - possible edpuzzle
2. What is the LHC? (Possible student engaged jigsaw or research activity)
3. Online game to stimulate LHC
4. Student report
Pedagogical Framework Score:
14/18
Skills Development:
5/10
Learning Skills: 4/4
Literacy Skills: 1/3
Life Skills: 0/3 OSOS Link Demonstrator 18 36 36 Demonstrator 19: Study Data from the Large Hadron Collider
Overview:
This exercise introduces students to CERN, the LHC
accelerator and the ATLAS and CMS experiments. It
explains the basics of the standard model of particle
physics and particle collisions. It explains what muons are
and how their angles can be measured through the use of
the HYPATIA event display or the CMS iSpy. Overview It mentions
what the expected distributions would look like and
prompts students to study and explain their results and
possible deviations for what is expected. OSOS Link Demonstrator 19
Metadata:
Age: 12 - 16
Duration: 2 hours
Equipment: PC with internet connection
Concepts introduced:
● Particle decays
● Distributions
● Histograms
Learning outcomes:
1. Students learn about CERN, LHC, ATLAS and CMS
2. They become familiar with the standard model of particle physics and particle collisions
3. They learn how to measure and analyse the angular distribution of muon tracks
4. They are asked to present their results and justify the outcome of their investigation
Prior knowledge:
● Structure of matter
● Fundamental forces
● Basic geometry
Learning intentions:
●
Understand the research being conducted at CERN, ATLAS and CMS
●
Understand how the LHC works
●
Learn what happens when particles collide in the LHC experiments
●
Study the distribution of muon track angles
Key activities:
1. Information about CERN, LHC, ATLAS, CMS
2. Explanation about the standard model and particle collisions
3. Investigation through the use of HYPATIA, iSpy
4. Presentation of student investigation results
Pedagogical Framework Score:
17/18
Skills Development:
8/10
Learning Skills: 4/4
Literacy Skills: 2/3
Life Skills: 2/3 Demonstrator 19: Study Data from the Large Hadron Collider Overview: This exercise introduces students to CERN, the LHC
accelerator and the ATLAS and CMS experiments. It
explains the basics of the standard model of particle
physics and particle collisions. It explains what muons are
and how their angles can be measured through the use of
the HYPATIA event display or the CMS iSpy. It mentions
what the expected distributions would look like and
prompts students to study and explain their results and
possible deviations for what is expected. 37 37 Demonstrator 20: The Magnetic Field and its Applications : The Magnetic Field and its Applications
students to CERN, the LHC
AS and CMS experiments. It
e standard model of particle physics
explains what muons are and how
sured through the use of the
r the CMS iSpy. It mentions what
ns would look like and prompts
plain their results and possible
pected. pp Overview: This exercise introduces students to CERN, the LHC
accelerator and the ATLAS and CMS experiments. It
explains the basics of the standard model of particle physics
and particle collisions. It explains what muons are and how
their angles can be measured through the use of the
HYPATIA event display or the CMS iSpy. It mentions what
the expected distributions would look like and prompts
students to study and explain their results and possible
deviations for what is expected. HYPATIA event display or the CMS iSpy. It mentions what
the expected distributions would look like and prompts
students to study and explain their results and possible
deviations for what is expected. OSOS Link Demonstrator 20
Metadata:
Age: 12 - 16
Duration: 2 hours
Equipment: PC with internet connection
Concepts introduced:
● Particle decays
● Distributions
● Histograms
Learning outcomes:
1. Students learn about CERN, LHC, ATLAS and CMS
2. They become familiar with the standard model of particle physics and particle collisions
3. They learn how to measure and analyse the angular distribution of muon tracks
4. They are asked to present their results and justify the outcome of their investigation
Prior knowledge:
● Structure of matter
● Fundamental forces
● Basic geometry
Learning intentions:
o
Understand the research being conducted at CERN, ATLAS and CMS
o
Understand how the LHC works
o
Learn what happens when particles collide in the LHC experiments
o
Study the distribution of muon track angles
Key activities:
1. Information about CERN, LHC, ATLAS, CMS
2. Explanation about the standard model and particle collisions
3. Investigation through the use of HYPATIA, iSpy
4. Presentation of student investigation results
Pedagogical Framework Score:
16/18
Skills Development:
5/10
Learning Skills: 3/4
Literacy Skills: 1/3
Life Skills: 1/3 OSOS Link Demonstrator 20 38 38 Demonstrator 21: The Pendulum
Overview:
This demonstrator introduces the concept of the pendulum and
the key concepts necessary to examine the properties of a
pendulum. The demonstrator is inquiry based and supports
students to develop controlled investigations on the different
variables that affect the period of a pendulum. They will be
looking at dependent and independent variables, and record data
that they will display and analyse. This demonstrator was designed as an introductory exercise for
the gravitational-wave package of demonstrators, and it
introduces the idea of a pendulum as a seismological filter for gravitational-wave detectors. Overview: This demonstrator introduces the concept of the pendulum and
the key concepts necessary to examine the properties of a
pendulum. The demonstrator is inquiry based and supports
students to develop controlled investigations on the different
variables that affect the period of a pendulum. They will be
looking at dependent and independent variables, and record data
that they will display and analyse. This demonstrator was designed as an introductory exercise for
the gravitational-wave package of demonstrators, and it
introduces the idea of a pendulum as a seismological filter for gravitational-wave detector OSOS Link Demonstrator 21
Metadata:
Age: > 12
Duration: 2 hours
Equipment: PC with internet connection and
basic lab equipment
Concepts introduced:
● Pendulum
● Period
● Frequency
● Gravitational wave detector
Learning outcomes:
1. To introduce students to the concept of a pendulum. 2. To investigate the relationships between the variables that affect the period of a pendulum
3. To graph and analyse data and form conclusions about how a pendulum operates. Prior knowledge:
●
Types of variables
●
Graphing
Learning intentions:
o
Define the following: pendulum, cycle/oscillation, frequency, period, variable
o
List examples of everyday pendulums
o
Develop a method for an investigation
o
Identify the types of variables
o
Graph the results of an investigation
o
Form conclusions based on the results collected
Key activities:
1. Pasta activity or alternative? 2. Design a basic pendulum
3. Plan and carryout investigation and collection of data
4. Graph results
5. Final Report
Pedagogical Framework Score:
16/18
Skills Development:
10/10
Learning Skills: 4/4
Literacy Skills: 3/3
Life Skills: 3/3 OSOS Link Demonstrator 21
Metadata:
Age: > 12
Duration: 2 hours
Equipment: PC with internet connection and
basic lab equipment
Concepts introduced:
● Pendulum
● Period
● Frequency
● Gravitational wave detector
Learning outcomes:
1. To introduce students to the concept of a pendulum. 2. To investigate the relationships between the variables that affect the period of a pendulum
3. To graph and analyse data and form conclusions about how a pendulum operates. Prior knowledge:
●
Types of variables
●
Graphing
Learning intentions:
o
Define the following: pendulum, cycle/oscillation, frequency, period, variable
o
List examples of everyday pendulums
o
Develop a method for an investigation
o
Identify the types of variables
o
Graph the results of an investigation
o
Form conclusions based on the results collected
Key activities:
1. Pasta activity or alternative? 2. Design a basic pendulum
3. Demonstrator Initial Teacher Training Workshops Demonstrator Initial Teacher Training Workshops Demonstrator Initial Teacher Training Workshops A range of methodologies were employed at initial
teacher training workshops to evaluate and
further develop the final demonstrators. Through
a series of pilot teacher training workshops
delivered by project partners between September
2019 and March 2020, data was collected and
analysed to evaluate the effectiveness of the
demonstrators. for
Figure 1 Initial Teacher Training event in Ireland 15 initial teacher training workshop events were
held by the project partner countries (Ireland,
Greece, Italy, France and Portugal). The general
format of the workshops was an introduction to the
project showing the FRONTIERS website and Facebook
page, an overview of the project framework followed by an examination of at least two
demonstrators. The workshop facilitators usually included a researcher in the field of physics
pertinent to the demonstrators explored and an education and outreach expert with a physics
background. Figure 1 Initial Teacher Training event in Ireland Figure 1 Initial Teacher Training event in Ireland DCU produced guidelines for conducting the face-to-face training events, with partners delivering
the presentations and workshop content in the native language of participants. The objective of the
workshop /multiplier events was to ensure that teachers could implement at least one demonstrator
in their classroom after the workshop. DCU produced guidelines for conducting the face to face training events, with partners delivering
the presentations and workshop content in the native language of participants. The objective of the
workshop /multiplier events was to ensure that teachers could implement at least one demonstrator
in their classroom after the workshop. There were slight variations in each
workshop with regard to the duration as
well as the mode of exploration of the
demonstrators. The recommended
workshop duration was 4 hours minimum. There were slight variations in each
workshop with regard to the duration as
well as the mode of exploration of the
demonstrators. The recommended
workshop duration was 4 hours minimum. Figure 2 Initial teacher training participants in Greece The initial teacher training workshops
were conducted with 303 science and
physics teachers across the partner
countries, with teachers from primary,
secondary education as well as in-training
teachers. Figure 2 Initial teacher training participants in Greece Overview: Plan and carryout investigation and collection of data
4. Graph results
5. Final Report
Pedagogical Framework Score:
16/18
Skills Development:
10/10
Learning Skills: 4/4
Literacy Skills: 3/3
Life Skills: 3/3 OSOS Link Demonstrator 21 Skills Development:
10/10 39 39 Pre and Post Workshop Questionnaire Question 5
I believe that I can motivate students to learn the
topics of Nobel Prize Physics. I believe that I can motivate students to learn the
topics of Nobel Prize Physics. The demographic data collected from teachers after following the proper consent procedures
included: country, gender and age of teachers. The demographic data collected from teachers after following the proper consent procedures
included: country, gender and age of teachers. Pre and Post Workshop Questionnaire The teachers completed a short pre-questionnaire in hard-copy at the start of the vision building
workshop and a post-questionnaire in hard-copy was completed at the end of the workshop. The pre
and post questionnaires, mainly addressed teachers’ motivation and self-efficacy. All questions were
answered through a 1-10 Likert scale. Likert scale questions encourage respondents to point to the
extent to which they agree or disagree with a particular statement. The advantage of the Likert scale
in this case was that it provided useful quantifiable data with regards teachers’ opinions and
perceptions around bringing Nobel Prize Physics topics into the classroom. 40 40 Pre-Training Questions
Post-Training Questions
Question 1
I appreciate the value of teaching Nobel Prize
Physics. The content of the learning event was
relevant/useful to my needs as a science teacher. Question 2
I am confident with the subject content of Nobel
Prize Physics. I am confident of the subject content of Nobel
Prize Physics. Question 3
I am interested in integrating Nobel Prize Physics
in my science classroom. The learning event has motivated me to further
explore Nobel Prize Physics with a view to
integrating it into the science class. Question 4
There are useful resources available (online) to
teach about Nobel Prize Physics. I can see the potential of applying Nobel Prize
Physics in the teaching of science. Question 5
I believe that I can motivate students to learn the
topics of Nobel Prize Physics. I believe that I can motivate students to learn the
topics of Nobel Prize Physics. The demographic data collected from teachers after following the proper consent procedures
included: country, gender and age of teachers. Pre-Training Questions
Post-Training Questions
Question 1
I appreciate the value of teaching Nobel Prize
Physics. The content of the learning event was
relevant/useful to my needs as a science teacher. Question 2
I am confident with the subject content of Nobel
Prize Physics. I am confident of the subject content of Nobel
Prize Physics. Question 3
I am interested in integrating Nobel Prize Physics
in my science classroom. The learning event has motivated me to further
explore Nobel Prize Physics with a view to
integrating it into the science class. Question 4
There are useful resources available (online) to
teach about Nobel Prize Physics. I can see the potential of applying Nobel Prize
Physics in the teaching of science. Questionnaire Findings We set out
could improve teachers’ self-efficacy in teachi
with the subject content (Question 2) and the
A th
i bl
d t t
Figure 3 Initial teacher training in Italy pre-questionnaire it is evident that prior to the workshops, teachers seemed to appreciate the value
of teaching Nobel Prize Physics (Mean = 8.31, SD = 1.69), and they were interested in integrating
Nobel Prize Physics in their science classroom (Mean = 8.34, SD = 1.57). Participants seemed
marginally confident with the subject content of Nobel Prize Physics (Mean = 6.02, SD = 2.46), and
were quite confident that they could motivate students to learn the topics of Nobel Prize Physics
(Mean = 7.63, SD = 1.60). Finally, the sample identifies that they were marginally confident that
there are useful resources available online to teach about Nobel Prize Physics (Mean = 6.25, SD =
1.95). Following the workshop training, participants completed a post-intervention questionnaire to
compare their confidence and attitudes, gauging the impact of the training. The pre and post
questionnaires completed by each teacher were matched. Following the workshop training, participants completed a post-intervention questionnaire to
compare their confidence and attitudes, gauging the impact of the training. The pre and post
questionnaires completed by each teacher were matched. The results of the post-questionnaire indicate that the participants found the content of the
workshops were relevant and useful to their needs as science teachers (Mean = 8.49, SD = 1.63). After the workshop participants were motivated to further explore Nobel Prize Physics with a view
to integrating it into the classroom (Mean = 8.52, SD = 1.60) and they can see the potential of
applying Nobel Prize Physics in the teaching of science (Mean = 8.34, SD = 1.51). Participants were
quite confident with the subject content of Nobel Prize Physics (Mean = 7.30, SD = 1.93), and
confident that they could motivate
students to learn the topics of
Nobel Prize Physics (Mean = 8.25,
SD = 1.51). Figure 3 Initial teacher training in Italy It is evident that the teachers’
confidence in the subject content
as well as confidence that they
could motivate students to learn
the topics of Nobel Prize Physics
had increased due to their
participation in the training
workshop. This increase will be
further investigated in the following Figure 3 Initial teacher training in Italy Figure 3 Initial teacher training in Italy section. section. Self-Efficacy One of the stated aims of the FRONTIERS project was to empower teachers in delivering Nobel prize
winning physics in the classroom. We set out to investigate whether the FRONTIERS workshops
could improve teachers’ self-efficacy in teaching Nobel prize Physics. This refers to their confidence
with the subject content (Question 2) and their capability to motivate students (Question 5). As the same variables are measured at two points in time with the same group (pre and post
workshop), paired sample t-tests were conducted to discover whether there are statistically
significant differences between means. Questionnaire Findings After the workshop participants were motivated to further explore Nobel Prize Physics with a vie
to integrating it into the classroom (Mean = 8.52, SD = 1.60) and they can see the potential of
applying Nobel Prize Physics in the teaching of science (Mean = 8.34, SD = 1.51). Participants wer
quite confident with the subject content of Nobel Prize Physics (Mean = 7.30, SD = 1.93), and
confident that they could motiva
students to learn the topics of
Nobel Prize Physics (Mean = 8.2
SD = 1.51). It is evident that the teachers’
confidence in the subject conte
as well as confidence that they
could motivate students to lear
the topics of Nobel Prize Physic
had increased due to their
participation in the training
workshop. This increase will be
further investigated in the follow
section. Self-Efficacy
One of the stated aims of the FRONTIERS project was to empower teachers in delivering Nobel p
winning physics in the classroom. We set out to investigate whether the FRONTIERS workshops
could improve teachers’ self-efficacy in teaching Nobel prize Physics. This refers to their confiden
with the subject content (Question 2) and their capability to motivate students (Question 5). A th
i bl
d t t
i t i ti
ith th
(
d
t
Figure 3 Initial teacher training in Italy pre-questionnaire it is evident that prior to th
of teaching Nobel Prize Physics (Mean = 8.31,
Nobel Prize Physics in their science classroom
marginally confident with the subject content
were quite confident that they could motivate
(Mean = 7.63, SD = 1.60). Finally, the sample i
there are useful resources available online to
1.95). Following the workshop training, participants
compare their confidence and attitudes, gaug
questionnaires completed by each teacher we
The results of the post-questionnaire indicate
workshops were relevant and useful to their n
After the workshop participants were motivat
to integrating it into the classroom (Mean = 8
applying Nobel Prize Physics in the teaching o
quite confident with the subject content of No
section. Self-Efficacy
One of the stated aims of the FRONTIERS proj
winning physics in the classroom. Questionnaire Findings Of the 303 workshop participants, 230 (76%) completed both the pre and post training
questionnaires. Of the 230 participant who completed both the pre and post training questionnaire, 105 were male
and 123 were female. Two participants did not indicate gender. The mean age of participants was 44
(SD=13), with the age of participants ranging from 19 to 64. The youngest of the participants were
Initial Teacher Education students from DCU, Ireland. A breakdown of ages and gender can be found
below. Prior to undertaking the pilot training, participants were asked to complete a pre-questionnaire to
explore the confidence and attitude of teachers with frontier science. From the data collected in the
0
10
20
30
40
50
60
70
80
<25
25-34
35-44
45-54
55-64
Age and Gender of Participants
Total
Male
Female 0
10
20
30
40
50
60
70
80
<25
25-34
35-44
45-54
55-64
Age and Gender of Participants
Total
Male
Female Prior to undertaking the pilot training, participants were asked to complete a pre-questionnaire to
explore the confidence and attitude of teachers with frontier science. From the data collected in the 41 41 pre-questionnaire it is evident that prior to the workshops, teachers seemed to appreciate the va
of teaching Nobel Prize Physics (Mean = 8.31, SD = 1.69), and they were interested in integrating
Nobel Prize Physics in their science classroom (Mean = 8.34, SD = 1.57). Participants seemed
marginally confident with the subject content of Nobel Prize Physics (Mean = 6.02, SD = 2.46), an
were quite confident that they could motivate students to learn the topics of Nobel Prize Physics
(Mean = 7.63, SD = 1.60). Finally, the sample identifies that they were marginally confident that
there are useful resources available online to teach about Nobel Prize Physics (Mean = 6.25, SD =
1.95). Following the workshop training, participants completed a post-intervention questionnaire to
compare their confidence and attitudes, gauging the impact of the training. The pre and post
questionnaires completed by each teacher were matched. The results of the post-questionnaire indicate that the participants found the content of the
workshops were relevant and useful to their needs as science teachers (Mean = 8.49, SD = 1.63). Self-Efficacy and Teacher Age There was a broad distribution of ages in our sample; from pre-service science teachers to teachers
with more than 30 years of experience. To explore possible relationship between age of participants
and the impact of the training workshop on participants, a Single Factor ANOVA was undertaken of
both question 2 and question 5 results, considered as relevant to teacher self-efficacy. The participants were divided into five age categories: under 25, 25-34, 35-44, 45-54 and 55-64. The
rationale for the inclusion of the 25 category was to measure the impact of the intervention with in-
training and recently qualified teachers. The score difference from the pre and post training workshop questionnaires were analysed for
questions 2 and 5. Question 5: I believe that I can motivate students to learn the topics of Nobel Prize Physics. Question 5 responses, referring to participant capability to motivate students to learn about Nobel
prize physics, were also found to have a statistically significant (p<.01) relationship between pre
(Mean = 7.63) and post (Mean = 8.25). The 8.12% increase in mean scores had a positive strength of
association considered large with a Pearson Correlation of 0.614. and post (Mean = 7.30) workshop mean scores. The 21.26% increase in mean scores had a positive
strength in association considered large with a Pearson Correlation of 0.724. and post (Mean = 7.30) workshop mean scores. The 21.26% increase in mean scores had a positive
strength in association considered large with a Pearson Correlation of 0.724. Question 2: I am confident with the subject content of Nobel Prize Physics. Question 2 responses, referring to participant confidence with the subject content of Nobel prize
physics, were found to have a statistically significant (p<.01) relationship between pre (Mean = 6.02) 42 42 42 A second Single Factor ANOVA found a statistically significant (p<0.01) relationship between age and
reported increase in confidence post workshop training. All age groups were found to have reported
an average increase in confidence with the subject matter of Nobel prize winning physics. The Under
25 and 25-34 age groups saw the largest reported increase in confidence, with an average increase
of more than 2 points on a 10-point scale (2.03 and 2.11 respectively). The group with the lowest
reported increase in confidence following the training was the 55-64 category, with an average
increase of less than 1 point on a 10-point scale. Anova: Single Factor
Reported increase in teacher confidence with subject matter by age
SUMMARY
Groups
Count
Average
Variance
Under 25
38
2.0263
5.9182
25-34
18
2.1111
3.0457
35-44
44
1.3409
1.8113
45-54
72
1.1111
2.6071
55-64
58
0.7068
1.3336
ANOVA
Source of Variation
SS
df
MS
F
P-value
F crit
Between Groups
54.864
4
13.716
5.0611
0.0006
2.4117
Within Groups
609.76
225
2.7100
Teacher confidence motivating students to learn about Nobel Prize winning physics Anova: Single Factor
Reported increase in teacher confidence with subject matter by age Teacher confidence motivating students to learn about Nobel Prize winning physics Question 5 of the pre and post training workshop questionnaire measured the confidence that
teachers had in their abilities to motivate students to learn about Nobel prize winning physics. An
initial analysis of variance found a statistically significant relationship (p<0.01) between the age of
the participants and their reported abilities to motivate students. The difference in average scores
between age groups was far narrower than the previous analysis. The lowest average score by age
group was the under 25s (6.76 on a 10-point scale), and the highest being the 55-64 age group (7.96
on a 10-point scale). Teacher Confidence with subject of Nobel Prize winning physics Question 2 of the pre and post training workshop questionnaire measured the confidence of
participating teachers with the subject of Nobel prize winning physics. The initial ANOVA analysis of
the reported confidence scores of teachers found a statistically significant relationship (p<0.01)
between the age of teachers and their reported confidence with Nobel prize winning physics. The
lowest average confidence scores were reported amongst the under 25 age category, while the
highest was reported amongst the 55-64 category. 43
0
1
2
3
4
5
6
7
8
Under 25
25-34
35-44
45-54
55-64
Reported Confidence with subject of Nobel prize
winning physics 0
1
2
3
4
5
6
7
8
Under 25
25-34
35-44
45-54
55-64
Reported Confidence with subject of Nobel prize
winning physics Reported Confidence with subject of Nobel prize
winning physics 43 Motivation of Teachers The high levels of motivation observed in teachers participating in the training workshops were
noted by several facilitators. Discussions at training workshops in Greece showed that teachers’
motivation was “to receive training in order to enrich their educational practice and connect with
cutting edge research in physics.” This eagerness to improve educational practice and knowledge of
modern physics was evident in many workshop participants. Despite the motivation and enthusiasm of teachers, it was reported at the beginning of one
workshop in Greece that despite their efforts, students did not seem interested in physics. Following workshops, many teachers reported that they could now see the potential of developing
Nobel Prize Physics in the teaching of science. In Ireland it was stated by teachers that the training
event had “motivated them to further explore Nobel Prize Physics with a view to integrating it into
the science lessons.” relationship (p=0.57) between age and reported increase in confidence post workshop training. All
age groups were found to have reported an average score increase between pre and post training
questionnaires, although not as great as those responses to the previous question analysed. relationship (p=0.57) between age and reported increase in confidence post workshop training. All
age groups were found to have reported an average score increase between pre and post training
questionnaires, although not as great as those responses to the previous question analysed. Anova: Single Factor
Reported increase in teacher confidence to motivate students by age
SUMMARY
Groups
Count
Average
Variance
Under 25
38
0.3421
3.2581
25-34
18
0.7222
2.5653
35-44
44
0.6818
2.2219
45-54
72
0.7777
1.5273
55-64
58
0.5344
0.9549
ANOVA
Source of Variation
SS
df
MS
F
P-value
F crit
Between Groups
5.5066
4
1.3766
0.7329
0.57030
2.4117
Within Groups
422.58
225
1.8781 Anova: Single Factor
Reported increase in teacher confidence to motivate students by age Anova: Single Factor
Reported increase in teacher confidence to motivate students by age Teacher and Facilitator Feedback In addition to the quantitative data provided by the pre and post workshop questionnaires,
qualitative data was collected by the workshop facilitators. Teacher feedback from post workshop
discussions were reported as well as facilitator observations. This qualitative data was thematically
analysed, with the major identified themes discussed below. The Single Factor ANOVA of question 5, measuring the reported increase in confidence of teachers
to motivate students to learn about Nobel prize winning physics, did not find a statistically significant
6
6,2
6,4
6,6
6,8
7
7,2
7,4
7,6
7,8
8
8,2
<25
25-34
35-44
45-54
55-64
Teacher confidence in ability to motivate students to
learn about Nobel prize winning physics Teacher confidence in ability to motivate students to
learn about Nobel prize winning physics The Single Factor ANOVA of question 5, measuring the reported increase in confidence of teachers
to motivate students to learn about Nobel prize winning physics, did not find a statistically significant 44 44 44 Practical Nature of Workshops The teachers appreciated the practical nature of the session as it gave them time to explore a few
demonstrators during the session. It was widely acknowledged that the “hands-on” and practical
approach of the training workshops motivated teachers to participate more fully and
enthusiastically. It was also noted in Greece, the contrast in teacher engagement between the 45 45 practical “hands-on” elements of the workshops compared to the more theoretical, with teachers
seeming “less interested in demonstrators that didn't follow a hands-on approach.” seeming less interested in demonstrators that didn t follow a hands-on approach. Those workshops that incorporated a live virtual visit to a research facility also reported teachers
improved engagement and enthusiasm. Those workshops that incorporated a live virtual visit to a research facility also reported teachers
improved engagement and enthusiasm. Following workshops in Ireland, feedback from the participants indicated that the workshop was
very relevant to their needs, and it helped to develop further confidence in the topic. In addition,
participating teachers indicated that they “felt much more confident after the online session about
the subject content of Nobel Prize Physics.” Curriculum There were mixed feelings reported by teachers participating in workshops on how the
demonstrators could be incorporated into existing curriculums and within limited class contact
times. Teachers in Ireland stated that a number of the demonstrators could be used to support the existing
Earth and Science strand of the science curriculum at Junior Cycle (14–15-year-olds). In France, it was highlighted that cross curricular links could be made as school programmes now
include mandatory computer programming classes. Teachers reported that it could interesting to be
provided with data files and have to build some sort of analysis code in the classroom rather than
use ready-to-use black-box apps. Some teachers in France also
suggested that, rather than using the
demonstrator during normal classes,
they found it very suited for specific
"ateliers scientifiques" (scientific
workshops) that are already taking
place in French high schools. It was
explained that “during workshops
there are no programme constraints
and the students participating are
already the most motivated.” Figure 5 Initial teacher training in Portugal Figure 5 Initial teacher training in Portugal In other countries a common concern in implementing the demonstrators was the pressures and
limitations of the curriculums of individual countries. In Italy it was raised that “given the limited
time available at school, would not be easy to dedicate 2 hours to a single demonstrator.” This
concern was echoed by participants in Greece, with teachers “reluctant to implement scenarios that
are outside the curriculum.” In other countries a common concern in implementing the demonstrators was the pressures and
limitations of the curriculums of individual countries. In Italy it was raised that “given the limited
time available at school, would not be easy to dedicate 2 hours to a single demonstrator.” This
concern was echoed by participants in Greece, with teachers “reluctant to implement scenarios that
are outside the curriculum.” It was suggested that a support to teachers would be mapping each of the demonstrators to national
curriculums. The ability of teachers to edit the demonstrators to their own needs was a major plus
for their applicability. The mapping of the demonstrators to target age groups was another topic of discussion across
workshops. ready-to-use tools and said that often they have to create their own material. The general concept
of providing them with demonstrators was very welcome. ready-to-use tools and said that often they have to create their own material. The general concept
of providing them with demonstrators was very welcome. With the feedback being generally very positive, it was noted at several workshops a desire by
teachers to engage in further detailed training in individual demonstrator. A workshop participant in
Greece suggested “systematic training one demonstrator at a time to have time to assimilate
content.” Some teachers also proposed additional training to improve their own knowledge in
modern physics. This was evident in Portugal, where several teachers said that they would need help
to implement the demonstrators as they “aren’t at ease with modern physics.” Demonstrators and Pedagogical Framework The demonstrators were received very
positively by participating teachers across
the partnership workshop events. In
France, the feedback was very positive
with teachers “convinced that
demonstrators can be brought to the
classroom.” Figure 4 Initial teacher training in France One of the most commonly reported
benefits for teachers was that the
demonstrators were designed using the
language of educators and an enquiry-
based learning approach. One Greek
participant commented that other
outreach activities that she had joined
would “just present content but don’t speak
the language of the educator and don’t offer a means to do them in the classroom.”
Figure 4 Initial teacher training in France Figure 4 Initial teacher training in France The inquiry-based learning approach adopted by the project was noted as being of great interest and
benefit to teachers and students. It was noted in Portugal that the “inquiry methodology was very
interesting because you feel that most teachers value such experience.” Teachers in France reported
inquiry-based learning is not only suited for their teaching but encouraged by National Education. One participant at a workshop in France stated “the most important point is to give a taste of the
scientific method and approach, since science is very often taught in class in a non-scientific fashion.”
The demonstrators in their content and design facilitated bringing the scientific approach to the
classroom in both its content and pedagogical approach. In Ireland, several participants commented on that the demonstrators encouraged students to
undertake research in the classroom using scientific language through the inquiry-based learning
model. One participant highlighted the demonstrator meant “students will be using scientific
language and interpreting real graphs, using graphs in real life.” The inquiry-based learning
approach to delivering content was identified as encouraging students to employ “critical thinking,
problem solving, justifying reasoning” through “practical investigation.” The demonstrators as a teaching resource were welcomed by teachers, with many reporting a lack
of physics education resources available. In general, participating teachers were not aware of online 46 46 Curriculum In Ireland it was felt that “many of the demonstrators are more appropriate for the
Leaving Certificate (17-18 year olds) syllabus.” This was the feeling among participants in France too,
who found that content may be “too complicated for the average student (except maybe last year of
high-school)”. 47 47 47 Teachers in Ireland suggested that less able students may struggle with some of the more difficult
concepts and “abstract thinking” covered in the demonstrators. It was suggested that “less able
students may need background lessons.” There was interest in participants to apply some of the demonstrators to younger school class
groups, including primary school and early high school classes. In was suggested by participants in
Portugal that demonstrators could be simplified for younger age groups. Teacher Professional Development The FRONTIERS Project undertook a range of teacher professional development events throughout
the project lifetime. As per the original project proposal, multiplier events had been planned as
FRONTIERS Visionary Workshops (E1-6) and FRONTIERS Practice Reflection Meetings (E7-12). As a
result of adjustments made to the project activities due to the Covid-19 pandemic, project partners
held professional development events that facilitated the quality engagement of teachers and
students within the revised scope of the project. These events included Teacher Training workshops,
Master Classes and Virtual Visits to large-scale research facilities. A series of in-person FRONTIERS International Training Courses/Summer Schools (C1-3) and an
International Symposium (E13) had also been outlined in the original project proposal. As
international travel and in-person meetings were heavily restricted, a series of online International
e-Schools were hosted by project partners. These International e-Schools facilitated hundreds of
teachers to learn about the FRONTIERS Demonstrators, collaborate with other teachers as well as
physics researchers to develop class plans, and join virtual visits to large-scale research facilities. The impact and effectiveness of these teacher training events are evaluated in the following
sections. Initial Teacher Training Workshops 15 initial teacher training workshops were held by the project partner countries (Ireland, Greece,
Italy, France and Portugal). The objective of these workshop multiplier events was to ensure that
teachers could implement at least one demonstrator in their classroom after the workshop. The
recommended workshop duration was 4 hours minimum. DCU produced guidelines for conducting
the face-to-face training events, with partners delivering the presentations and workshop content in
the native language of participants. The workshops were offered in the teachers’ native language. The general format of the workshops
was an introduction to the project showing the FRONTIERS website and Facebook page, an overview
of the project framework followed by an examination of at least two demonstrators. The workshop
facilitators usually included a researcher in the field of Physics pertinent to the particular
demonstrators explored and an education and outreach expert with a Physics background. The initial teacher training workshops were conducted with 303 science and physics teachers across
the partner countries, with teachers from primary, secondary education as well as in-training
teachers. The evaluation findings of the initial teacher training events were explored in the previous
section. Technology in the Classroom Teachers generally felt that equipment and technology available in schools could be a barrier to
engagement with the demonstrators. In Portugal, some teachers stated that “the need for
computers can be a drawback” as they are not always accessible or available. In France there was a concern that the demonstrators were strictly online, as good Wi-Fi connection
seems to be an issue for some schools and educators. 48 48 International e-Schools The FRONTIERS Project ran three online training courses for science teachers in the form of
International e-Schools. Hosted through Zoom, these synchronous online training events targeted
science teachers who were motivated to introduce Nobel prize winning physics in their classroom. The lesson learned and insights gained from the initial teacher training workshops events informed
the design of the International e-School series. Due to the COVID 19 pandemic, the scale of implementation of the FRONTIERS Demonstrators
initially envisaged for the project was not possible; school were closed to students for large periods 49 49 49 and online learning meant drastic changes in teaching and learning activities. For this reason, the
Frontier project partners decided to focus on engaging with, training and community building of
motivated science teachers through online international training events. The objectives of these online training events were: he objectives of these online training events were: ● Introduce participants to the FRONTIERS pedagogical design and the proposed activities for
bringing Nobel prize physics to the classroom. ● Visit to large scale research facilities in physics with the opportunity to interact with
researchers. ● Discuss of the merits and barriers of introducing Nobel Prize Physics to the Classroom and
presentation of the FRONTIERS methodology to achieve that. ● Provide hands on experience of using demonstrators and opportunities for teachers to
develop lesson plans. In response to the feedback gathered from teachers participating in the initial teacher training
workshops, emphasis was given to: In response to the feedback gathered from teachers participating in the initial teacher training
workshops, emphasis was given to: ● providing in-depth and detailed training to teachers on individual demonstrators ● facilitating teacher collaboration to adapt demonstrators to use with their students ● hosting participants on virtual visits to research facilities ● building a community of teachers motivated to bringing frontier physics to their student Both quantitative and qualitative data was collected to facilitate the evaluation of the International
e-School series. Quantitative data was collected using pre, and post questionnaires with participants. Round table discussions and focus groups were held with participants to gather qualitative data from
the participant perspective. The 3 online training courses that comprised the International e-Schools
are outlined below. Summer School July 2020 The FRONTIERS Project International Summer School 2020 took place over ten days between for the
13th and 24th of July 2020. A link to the agenda and relevant presentations is here, with an agenda
summary provided below. 50 teachers fully completed the Summer School 2020 receiving
certification. The first week of the Summer School 2020 included key-note speakers, introduction to the
FRONTIERS Project, presentation of demonstrators to teachers and formation of module-oriented
teacher working groups. The demonstrator modules explored each day included workshops on how
to bring Gravitational Waves, Astrophysics, High Energy Physics and Cosmology to the classroom. Participants were also given a virtual tour of the Virgo Interferometer in Italy. 50
Monday
Tuesday
Wednesday
Thursday
Friday
Introduction to
FRONTIERS
Key Notes Speakers
Virtual Visit to Virgo
Interferometer
Astrophysics in the
Classroom Module
●
Cloud Chamber
●
Relativistic Muons
●
Cosmic Rays
High Energy Physics in the
Classroom Module
●
Accelerating
Particles
●
Mass Energy
Equivalence
●
Z and Higgs bosons
Astrophysics in the
Classroom Module
●
Black holes
●
Alien Worlds
●
Exploring the Sun
●
Age of the galaxy
Gravitational Waves in
the Classroom Module
●
Build an
Interferometer
●
The Pendulum
●
Study Earthquakes
●
Discover Black
Holes 50
Monday
Tuesday
Wednesday
Thursday
Friday
Introduction to
FRONTIERS
Key Notes Speakers
Virtual Visit to Virgo
Interferometer
Astrophysics in the
Classroom Module
●
Cloud Chamber
●
Relativistic Muons
●
Cosmic Rays
High Energy Physics in the
Classroom Module
●
Accelerating
Particles
●
Mass Energy
Equivalence
●
Z and Higgs bosons
Astrophysics in the
Classroom Module
●
Black holes
●
Alien Worlds
●
Exploring the Sun
●
Age of the galaxy
Gravitational Waves in
the Classroom Module
●
Build an
Interferometer
●
The Pendulum
●
Study Earthquakes
●
Discover Black
Holes 50 Table 1: Summer School 2020 week 1 agenda summary
Week 2 of the Summer School 2020 introduced participants to ways in which Nobel prize winning
physics can be brought to life in the classroom and beyond, with working groups of participants
collaborating to develop their own lesson plans based on FRONTIERS Demonstrators. Summer School July 2020 Table 2: Summer School week 2 agenda summary
Monday
Tuesday
Wednesday
Thursday
Friday
Introducing Nobel Prize
Physics to the Primary
School
●
Playing with
Protons Initiative
●
Introduction to
CERN
●
Particles and their
Interactions
●
Q&A
REINFORCE Visionary
Workshop Day 1
●
Research
Infrastructure for
Citizens in Europe
●
Interactive Session
●
Zooniverse
●
Fishing for
Neutrinos
REINFORCE Visionary
Workshop Day 2
●
Citizen science and
school education
●
Gravitational Noise
Hunting in the
classroom
Working group activities
and evaluation
●
Final rehearsal of
working groups
●
Round Table
Discussions
●
Evaluation
●
Future Outlook
Final Presentations
●
Working group
presentation of
class materials
●
Astrophysics
●
Gravitational
Waves
●
High Energy Physics
Winter School 2020
The FRONTIERS Project Winter School 2021 took place over 6 days between the 29th of January and
the 7th of February 2021. To facilitate teachers attending during the normal academic year and to
avoid further class disruption, the Winter School took place over 2 weekends with meetings
scheduled between both weekends. In total there were 203 participants in flexible attendance, with
30 teachers chosen to participate in workshops groups to develop teaching resources. The Winter School followed a similar format as the previous Summer School. A link to the agenda
and relevant presentations is available here, with an agenda summary provided below. The first
weekend of the Winter School focused on introducing the participants to the FRONTIERS Project,
establishing working groups of participants, workshops on demonstrators and visiting research
facilities. The working groups used Google slides to collaborate on the development of lesson plans. There were scheduled times set aside between both weekends for participants to meet with experts Table 2: Summer School week 2 agenda summary
Monday
Tuesday
Wednesday
Thursday
Friday
Introducing Nobel Prize
Physics to the Primary
School
●
Playing with
Protons Initiative
●
Introduction to
CERN
●
Particles and their
Interactions
●
Q&A
REINFORCE Visionary
Workshop Day 1
●
Research
Infrastructure for
Citizens in Europe
●
Interactive Session
●
Zooniverse
●
Fishing for
Neutrinos
REINFORCE Visionary
Workshop Day 2
●
Citizen science and
school education
●
Gravitational Noise
Hunting in the
classroom
Working group activities
and evaluation
●
Final rehearsal of
working groups
●
Round Table
Discussions
●
Evaluation
●
Future Outlook
Final Presentations
●
Working group
presentation of
class materials
●
Astrophysics
●
Gravitational
Waves
●
High Energy Physics Summer School July 2020 ●
Large Hadron
Collider Data Table 1: Summer School 2020 week 1 agenda summary
●
Large Hadron
Collider Data Week 2 of the Summer School 2020 introduced participants to ways in which Nobel prize winning
physics can be brought to life in the classroom and beyond, with working groups of participants
collaborating to develop their own lesson plans based on FRONTIERS Demonstrators. Week 2 of the Summer School 2020 introduced participants to ways in which Nobel prize winning
physics can be brought to life in the classroom and beyond, with working groups of participants
collaborating to develop their own lesson plans based on FRONTIERS Demonstrators. Table 2: Summer School week 2 agenda summary
Monday
Tuesday
Wednesday
Thursday
Friday
Introducing Nobel Prize
Physics to the Primary
School
●
Playing with
Protons Initiative
●
Introduction to
CERN
●
Particles and their
Interactions
●
Q&A
REINFORCE Visionary
Workshop Day 1
●
Research
Infrastructure for
Citizens in Europe
●
Interactive Session
●
Zooniverse
●
Fishing for
Neutrinos
REINFORCE Visionary
Workshop Day 2
●
Citizen science and
school education
●
Gravitational Noise
Hunting in the
classroom
Working group activities
and evaluation
●
Final rehearsal of
working groups
●
Round Table
Discussions
●
Evaluation
●
Future Outlook
Final Presentations
●
Working group
presentation of
class materials
●
Astrophysics
●
Gravitational
Waves
●
High Energy Physics
Winter School 2020
The FRONTIERS Project Winter School 2021 took place over 6 days between the 29th of January and
the 7th of February 2021. To facilitate teachers attending during the normal academic year and to
avoid further class disruption, the Winter School took place over 2 weekends with meetings
scheduled between both weekends. In total there were 203 participants in flexible attendance, with
30 teachers chosen to participate in workshops groups to develop teaching resources. The Winter School followed a similar format as the previous Summer School. A link to the agenda
and relevant presentations is available here, with an agenda summary provided below. The first
weekend of the Winter School focused on introducing the participants to the FRONTIERS Project,
establishing working groups of participants, workshops on demonstrators and visiting research
facilities. The working groups used Google slides to collaborate on the development of lesson plans. There were scheduled times set aside between both weekends for participants to meet with experts
if they had any questions around the content of the demonstrators. Table 3: Winter School 2021 weekend 1 agenda summary Winter School 2020 Winter School 2020
The FRONTIERS Project Winter School 2021 took place over 6 days between the 29th of January and
the 7th of February 2021. To facilitate teachers attending during the normal academic year and to
avoid further class disruption, the Winter School took place over 2 weekends with meetings
scheduled between both weekends. In total there were 203 participants in flexible attendance, with
30 teachers chosen to participate in workshops groups to develop teaching resources. The FRONTIERS Project Winter School 2021 took place over 6 days between the 29th of January and
the 7th of February 2021. To facilitate teachers attending during the normal academic year and to
avoid further class disruption, the Winter School took place over 2 weekends with meetings
scheduled between both weekends. In total there were 203 participants in flexible attendance, with
30 teachers chosen to participate in workshops groups to develop teaching resources. The FRONTIERS Project Winter School 2021 took place over 6 days between the 29th of January and
the 7th of February 2021. To facilitate teachers attending during the normal academic year and to
avoid further class disruption, the Winter School took place over 2 weekends with meetings
scheduled between both weekends. In total there were 203 participants in flexible attendance, with
30 teachers chosen to participate in workshops groups to develop teaching resources. The Winter School followed a similar format as the previous Summer School. A link to the agenda
and relevant presentations is available here, with an agenda summary provided below. The first
weekend of the Winter School focused on introducing the participants to the FRONTIERS Project,
establishing working groups of participants, workshops on demonstrators and visiting research
facilities. The working groups used Google slides to collaborate on the development of lesson plans. There were scheduled times set aside between both weekends for participants to meet with experts
if they had any questions around the content of the demonstrators. Winter School 2020 Table 3: Winter School 2021 weekend 1 agenda summary Saturday 51 51 Welcome to the Winter School
The FRONTIERS Project
Inquiry Based Learning
Establishing working groups
Virtual Visit to CERN
Module 1: Gravitational Waves
●
Introduction
●
Demonstrators
●
Working Groups
Module 2: High Energy Physics
●
Introduction
●
Demonstrators
●
Working Groups
Virtual Visit to Virgo Interferometer
Module 3: Astrophysics and Cosmology
●
Introduction
●
Demonstrators
●
Working Groups
Module 4: Astroparticle Physics
●
Introduction
●
Demonstrators
●
Working Groups
Virtual visit to Pierre Auger Observatory
The second weekend of the Winter School showcased FRONTIERS best practice applications of Nobel
prize winning physics in primary and high schools. Inspired by these educational experiences,
participants were then given the opportunity to present their working group lesson plans developed
from the FRONTIERS Demonstrators. Module 3: Astrophysics and Cosmology
●
Introduction
●
Demonstrators
●
Working Groups
Module 4: Astroparticle Physics
●
Introduction
●
Demonstrators
●
Working Groups
Virtual visit to Pierre Auger Observatory The second weekend of the Winter School showcased FRONTIERS best practice applications of Nobel
prize winning physics in primary and high schools. Inspired by these educational experiences,
participants were then given the opportunity to present their working group lesson plans developed
from the FRONTIERS Demonstrators. Table 4: Winter School 2021 weekend 2 agenda summary
Saturday
Sunday
FRONTIERS Best Practice Experience
●
Primary Schools
●
High Schools
●
e-Twinning project
Working groups with mentors
Working groups’ presentation of educational materials adapted
from FRONTIERS Demonstrators to being to the classroom. FRONTIERS Next Steps Presentation
Winter School Final Remarks Table 4: Winter School 2021 weekend 2 agenda summary
Saturday
Sunday
FRONTIERS Best Practice Experience
●
Primary Schools
●
High Schools
●
e-Twinning project
Working groups with mentors
Working groups’ presentation of educational materials adapted
from FRONTIERS Demonstrators to being to the classroom. FRONTIERS Next Steps Presentation
Winter School Final Remarks Table 4: Winter School 2021 weekend 2 agenda summary Table 4: Winter School 2021 weekend 2 agenda summary Summer School 2021 The FRONTIERS Project International Summer School 2021 took place over five days between for the
12th and 16th of July 2021. A link to the agenda and relevant presentations is here, with an agenda
summary provided below. 25 teachers fully completed the Summer School 2021 receiving
certification. The FRONTIERS Project International Summer School 2021 took place over five days between for the
12th and 16th of July 2021. A link to the agenda and relevant presentations is here, with an agenda
summary provided below. 25 teachers fully completed the Summer School 2021 receiving
certification. The Summer School 2021 included key-note speakers, introduction to the FRONTIERS Project,
presentation of demonstrators to teachers and visits to large research institutes in physics. The
demonstrator modules explored each day included workshops on how to bring Gravitational Waves,
Astrophysics, High Energy Physics and Cosmology to the classroom. 52
Table 5: Summer School 2021 agenda summary
Monday
Tuesday
Wednesday
Thursday
Friday
Introduction to
FRONTIERS
Muon Lifetime at rest
and in flight
Cosmic Rays,
Muography and Citizen
Science
Virtual Visit to Pierre
Auger Observatory
Virtual visit to Virgo
The Pendulum and
Gravitational Wave
detection
Investigating the
Michelson
Interferometer
Gravitational Wave
Noise Hunting
An introduction to High
Energy Physics
FRONTIERS
Demonstrators on High
Energy Physics
Virtual Visit to ALICE
Experiment at CERN
Discussion
FRONTIERS
Demonstrators on
Astrophysics and
Cosmology
Introduction and live
observation with the
Faulkes Telescope
Discussion
FRONTIERS Best
Practices
Developing the
FRONTIERS Networks
Round Table Discussion
End of Summer School Table 5: Summer School 2021 agenda summary
Monday
Tuesday
Wednesday
Thursday
Friday
Introduction to
FRONTIERS
Muon Lifetime at rest
and in flight
Cosmic Rays,
Muography and Citizen
Science
Virtual Visit to Pierre
Auger Observatory
Virtual visit to Virgo
The Pendulum and
Gravitational Wave
detection
Investigating the
Michelson
Interferometer
Gravitational Wave
Noise Hunting
An introduction to High
Energy Physics
FRONTIERS
Demonstrators on High
Energy Physics
Virtual Visit to ALICE
Experiment at CERN
Discussion
FRONTIERS
Demonstrators on
Astrophysics and
Cosmology
Introduction and live
observation with the
Faulkes Telescope
Discussion
FRONTIERS Best
Practices
Developing the
FRONTIERS Networks
Round Table Discussion
End of Summer School Table 5: Summer School 2021 agenda summary 52 52 52 Data and Analysis The objectives of the evaluative analysis were threefold: 1. Had the FRONTIERS Project delivered high-quality online training on the demonstrators? 2. What impact had summer school on the development of teacher competencies and empo
implementation of FRONTIERS physics in the classroom? 3. Had the FRONTIERS Project established a community of teachers motivated to bringing frontier
physics to the classroom? Of the 50 teachers who successfully completed the Summer School 2020, 22 fully completed the pre,
mid and post engagement questionnaires on whose data quantitative analysis was undertaken. Of the 50 teachers who successfully completed the Summer School 2020, 22 fully completed the pre,
mid and post engagement questionnaires on whose data quantitative analysis was undertaken. In addition to the questionnaire, qualitative data was collected from participants in the form of a
focus group and round table discussions. These qualitative methods allowed the project partners to
understand the participant experience in greater depth. The scope of the focus group was to explore participant experience of the working group
collaboration opportunities with other teachers facilitated by the FRONTIERS Winter School. The
working group teacher collaborations had been run twice as part of the Summer School 2020 and
the Winter School 2021. This qualitative data collection and analysis from the focus group provided a
deeper understanding of the impact the project was having from the perspective of the teachers
involved. Five participants from the Winter School took part in the focus group which took place for 45
minutes over Zoom. Participants were from Poland, Romania, Italy and Greece, with two male and
three female participants. Round table discussions took place with participants at the end of each International e-School event. The purpose of these discussions was to capture data on the experience of participants through the
voices of participants as soon as possible, ensuring an accuracy and detail of their account. A joint analysis of these qualitative and quantitative findings was employed to evaluate the project
impact guided by the above stated objectives. 1. Had the FRONTIERS Project delivered high-quality online training on the demonstrators Teacher C stated, “I liked all
things…but if had to choose one
thing it would be the
collaboration with colleagues
far away from me.” The passion
and motivation of the
participants was summed up by
Teacher B who also said that the
sharing of ideas and
collaboration with other physics
teachers was the most
important part of the Winter
School “as it shows that even in
hard times we have the will and
energy to create amazing things
and to make this work.” Teacher C valued the collaborative nature of the Winter schools as “we saw
how our colleagues were thinking about these topics that we love.”
Figure 6 A screenshot from the virtual visit to CERN Teacher C stated, “I liked all
things…but if had to choose one
thing it would be the
collaboration with colleagues
far away from me.” The passion
and motivation of the
participants was summed up by
Teacher B who also said that the
sharing of ideas and
collaboration with other physics
teachers was the most
important part of the Winter
School “as it shows that even in
hard times we have the will and
energy to create amazing things
and to make this work.” Teacher C valued the collaborative nature of the Winter schools as “we saw
how our colleagues were thinking about these topics that we love.”
Figure 6 A screenshot from the virtual visit to CERN Figure 6 A screenshot from the virtual visit to CERN Figure 6 A screenshot from the virtual visit to CERN Figure 6 A screenshot from the virtual visit to CERN hard times we have the will and
energy to create amazing things
and to make this work.” Teacher C valued the collaborative nature of the Winter schools as “we saw
how our colleagues were thinking about these topics that we love.”
Figure 6 A screenshot from the virtual visit to CERN energy to create amazing things
and to make this work.” Teacher C valued the collaborative nature of the Winter schools as “we saw
how our colleagues were thinking about these topics that we love.” All the participants of the focus group felt that the collaboration with teachers and having the time
to work through questions that they had to solve in the working groups was very useful as it would
help them prepare for when their own students asked questions. 1. Had the FRONTIERS Project delivered high-quality online training on the demonstrators The quality of the online training provided to participants was evaluated in part through the
questionnaire response. Using a 5-point scale, questionnaire responses indicated participants were
interested and enjoyed the summer school (M=4.25, SD=0.698) Participants also reported the
training course was very valuable and useful to them (M=4.8, SD=0.48). These quantitative findings were reinforced by the qualitative findings from the focus group of
participants. The focus group participants also highlighted that the collaborative design of the
Winter School helped the teacher develop a deeper understanding of the content and educational
tool by allowing the professional explore and develop the resources together. Teacher C felt that
they understood the demonstrators from the presentations, “but after the collaboration I
understood many things much better as the other teachers gave many ideas about things that I
didn’t notice /think about before.” As a priority of the Summer School was to provide teachers with the opportunity to collaborate with
other teachers and the facilitators through the online platform, it was important to gather responses
on this aspect of the course. Respondents reported being satisfied with the level of collaboration 53 with other teachers (M=4.18, SD=1.01), and very satisfied with the level of collaboration they
experienced with course organisers (M=4.68, SD=0.72). with other teachers (M=4.18, SD=1.01), and very satisfied with the level of collaboration they
experienced with course organisers (M=4.68, SD=0.72). From the focus group findings, the standout element that participants enjoyed and felt that they
benefitted from most was the opportunity to collaborate with other teachers who share a passion
for physics. This was reflected in the comments of most of the focus group participants. The organisation of the International e-School was given much praise. Although it was generally felt
that participants would have benefitted from having face-to-face engagement, there were a number
of benefits to the online nature. It was acknowledged that there were benefits to participants having a virtual event. Teacher A said,
“I think it was much better than a face to face meeting, collaboration was easy using chat from
Google slides.” The online nature of the Winter School also facilitated virtual visits to research
facilities that would not have been accessible otherwise. Teacher A commented on the benefit of the
virtual visits, “visiting all the sites that I may never be able to visit”. Comparing the online Winter School to other face-to-face workshops and events that participants
had attended, some benefits and drawbacks were identified. Teacher B recalled a European Space
Agency workshop they attended and commented on how valuable it was “to make things together,
construct things and talk to each other and maybe talk to the other group at the next table.”
Although Teacher B felt that the Winter School was really well organised, “if face to face it would be
awesome. All the demonstrators and we
would have the chance to make the
constructions and collaborate and to be
closer to each other and communicate
in a better way.” Figure 7 Screenshot from International e-School The Winter School was the first time
that Teacher B had taken part in a
virtual workshop. While they said that
the Winter School was “great”, they
suggested that it could be done
“blended”. “A mix of face to face and
virtual would be better e.g. first part as
virtual workshop but the second time
after one or two months face to face.” Figure 7 Screenshot from International e-School 1. Had the FRONTIERS Project delivered high-quality online training on the demonstrators On average, participants experienced no difficulties in collaborating with other teachers during the
summer school (M=1.91, SD=1.06). On average, participants experienced no difficulties in collaborating with other teachers during the
summer school (M=1.91, SD=1.06). As the International e-Schools were held virtually, online collaborative tools had to be used to
facilitate the teacher engaging with the content and each other. This brought about several
challenges but also opportunities to work differently yet effectively. As the International e-Schools were held virtually, online collaborative tools had to be used to
facilitate the teacher engaging with the content and each other. This brought about several
challenges but also opportunities to work differently yet effectively. Although local internet connection issues for participants were mentioned, several of the
collaborative tools employed by the Frontier Project were subject to praise. The use of Zoom to host
the live events and sharing of resources by email was functional and accessible to all participants. In
the working groups, Google Slides received considerable praise. Teacher A explained how beneficial Google Slides was in helping with communication and
collaboration. “Google slides helped with the language barriers….. the use of chat in google slides
was excellent as we could communicate even if we were not present at the same time, whereas in
zoom the chat is all gone in chat but the chat on google slides was always there…” This praise was echoed by Teacher B when saying, “we had the chance to write what we think and
talk through the chat and express our ideas and organise collaboratively what we wanted to do. It
was a very convenient and easy way to collaboration…It was easily accessible to everyone.” 54 54 2. What impact had the International e-Schools had on the development of teacher
competencies and empowering implementation of FRONTIERS physics in the classroo competencies and empowering implementation of FRONTIERS physics in the classroom? own students about these topics…” This supported development of professional confidence and self-
efficacy was felt would be of great benefit to the teachers when delivering Nobel prize winning
physics content in their classrooms. own students about these topics…” This supported development of professional confidence and self-
efficacy was felt would be of great benefit to the teachers when delivering Nobel prize winning
physics content in their classrooms. All the participants of the focus group felt
that the collaboration with teachers and
having the time to work through questions
that they had to solve in the working
groups was very useful as it would help
them prepare for implementing the
demonstrator materials in their own
lessons and better prepare them for when
their own students asked questions in
class. Figure 8 International e-School participants The participants at the round table
discussion following Summer School 2021
revealed the training had improved the confidence of teachers in delivering frontier science in the
classroom in a dynamic and meaningful way. Figure 8 International e-School participants Participant 4 stated that, “Physics is a nice subject but the blackboard is not enough to give the
students the pleasure of intellectual discoveries.” They felt that the “practical aspect of FRONTIERS
resources will instil in students the ability to ask questions and to explore.” The feeling that the FRONTIERS Demonstrators and International e-Schools would provide teachers
with the tools, knowledge and confidence to bring frontier science to the classroom was held
amongst many contributing participants. Participant 5, who had taken part in two of the
International e-Schools, said “some of the topics are very difficult for the teacher but these
demonstrators help. I am starting to use the demonstrators in the classroom.” She had tried to
introduce these topics previously but in a “theoretical way”. Using what they had learned through
the International e-Schools, they had started “to introduce the topics in a practical way with the
animations and videos that are part of the demonstrators”. As a result, she and her students “feel
like working like a scientist.” The online nature of the International e-School provided Participant 7 with the knowledge, tools and
opportunity to engage students with frontier science. They had attended the summer school event
to help them “introduce modern physics into the classroom”. They found great benefit from “doing
observation online and having a chance to visit CERN and other facilities. I will use all of these
materials to teach my students.” competencies and empowering implementation of FRONTIERS physics in the classroom Self-Efficacy of participating teachers in teaching Nobel prise physic was measured in pre, mid and
post summer school questionnaires. A significant increase was recorded in the average Self-Efficacy
score from pre (M=3.8), mid (M=4.0) and post (M=4.5). Respondents also reported being far more aware of resources available to them to support bringing
Nobel prize winning physics to the classroom. This correlated with a measurable increase in the
reported feasibility of implementing modern physics in the classroom. In measuring the confidence and self-perceived competence of participating teachers, positive
outcomes were recorded. The confidence of the participant teachers with content knowledge of
modern physics increased comparing pre (M=3.2) and post (M=4.0) training scores. The post
training, participants reported feeling competent to design educational content relating to modern
physics (M=4.2, SD 0.635). From the focus group findings, Teacher E explained that working with other colleagues to develop
resources and “meeting with mentors was very helpful to strengthen our confidence to talk to our 55 One of the main reasons given for taking part in the Winter School by some of the focus group
participants, was that they wanted the opportunity to learn from teachers from other countries. Learning about different national physics curriculums was very valuable. Teacher C stated “The main reason I wanted to be part of the e-Winter school was it was very
interesting and exciting to collaborate and to listen to what teachers from other countries had to
say.” Teacher E echoed this, explaining that the opportunity to compare curriculums was an
important learning experience. “Working with teachers from other countries in the working group
allowed us to … compare curriculums from different countries.” Teacher E was surprised to find how
different each national curriculum was with areas seemingly developed to varying degrees. Although there were a lot of differences discovered between national curriculums, the teachers
found it interesting and exciting working to find common ground. Fertile creative ground was found
between participants as Teacher B explained, “we all found something in common and managed to
collaborate and know that we can find ways if we want even if you have different curriculums and
this is something we can expand through collaboration with schools from different countries…” The differences in national curriculums was overcome by the passion and motivation of the teachers
involved, described Teacher A. “We are all teachers, we have the same goals in all countries. We
want to teach the students and help them to learn. It was interesting and very useful to see what
levels of study there is in other countries.” When asked if they would be interested in participating in follow-up meetings and/or activities, the
response from participants were very positive (M=4.5, SD=0.80). There was strong interest in participating in follow-up meeting and activities in focus group and
round table discussions. Participant 9 of the round table discussion hoped that the “collaboration
continues as it was a great experience for me and my students.” 3. Has the FRONTIERS Project established a community of teachers motivated to bringing frontier
physics to the classroom? Following the completion of the summer school, participants on average reported being very
interested in implementing the FRONTIERS Demonstrators with their students this year (M=4.82,
SD=0.39). Participants were also very interested in collaborating with colleagues to create e-twinning
projects (M=4.64, SD=0.73). 56 56 The FRONTIERS Project invited teachers trained in Gravitational Wave Astronomy in the framework
of FRONTIERS to perform virtual visits with their students. As part of this invitation, teachers were
offered educational resources for an introduction of the topic to the classroom and an assessment
framework and tools to assess the impact and educational merits of the visit. In total, 7 time slots
were reserved for these virtual visits, in which more than 617 students and 24 teachers. The
participants came from 24 schools in 8 countries (Greece (15); Italy (2); Romania (2); France (1);
Pakistan (1); Portugal (1); Bangladesh (1); Austria (1)). Virgo, the largest Gravitational Wave detector in Europe, has been designed and built by a
collaboration between the French Centre National de la Recherche Scientifique (CNRS) and the
Italian Instituto Nazionale di Fisica Nucleare (INFN). It is now operated and improved in Cascina, a
small town near Pisa on the site of the European Gravitational Observatory (EGO), by
an international collaboration of scientists from France, Italy, the Netherlands, Poland, and Hungary. Virtual visits to Virgo were streamed to students live via Zoom. They were moderated by an
education researcher of Ellinogermaniki Agogi and the field work was carried out by researchers and
support staff of EGO/Virgo. They were filmed through a mobile phone streaming directly to the
Zoom call which was led by an EGO/Virgo researcher, who accompanied the visitors through various
areas of the Virgo experiment. The structure of the tour allowed the visitor to explore Virgo and find
out about its mission: from the theory of how interferometry works and how it can be used to listen
to the cosmos to the extraordinary experimental challenges that need to be overcome in order to
accomplish that. The visits lasted about 90-120 minutes and was composed of five parts: •
Introductory seminar •
Exhibits located in the hall of the EGO Main Building After each of these the visitors were encouraged to ask questions, with 5 minutes dedicated to the
questions after each episode. To assess the episodes of the visit, a dedicated evaluation instrument
was developed by the EA project partners. A pre and post student questionnaire was developed to measure the effectiveness of the virtual
visits in impacting on key performance indicators. A link to each of the questionnaires can be found
here: Pre-Questionnaires and Post-Questionnaires. Virtual Visits Figure 9 Flyer for International e-School Summer
2021 To directly engage students with Nobel prize winning
physics and the FRONTIERS Project, a series of virtual visits
to large-scale research facilities were facilitated by project
partners. In total 1,100 students and 192 teachers took
part in 11 virtual visits, allowing access to real-world
physics experiments and the research scientists that work
there. The virtual visits undertaken included a visit to the ATLAS
experiment at CERN in January 2021 organised by project
partners IASA and EA. 150 students participated with it
also broadcast live on Facebook. In May 2021, IASA
organised 2 virtual visits were arranged to the ALICE
experiment at CERN, in which 333 students from 13
schools took part. DCU organised a virtual visit to the ALICE experiment in May for 20 science
teachers in Ireland. In addition, EA organised a series of 7 virtual visits to the VIRGO experiment,
from which pre and post visit data was gathered from teachers and students. These visits and the
analysis of data gathered from participated are discussed below. Figure 9 Flyer for International e-School Summer
2021 57 57 57 offered an introductory presentation about Gravitational Waves by their teachers on average three
days before the visit and answered the pre- questionnaires after that. The post questionnaires were
delivered within 1 week from the completion of the visit. offered an introductory presentation about Gravitational Waves by their teachers on average three
days before the visit and answered the pre- questionnaires after that. The post questionnaires were
delivered within 1 week from the completion of the visit. In order to investigate students’ familiarity with related concepts we performed a non-parametric
Related Samples Wilcoxon Signed Rank test for each vocabulary item to compare the pre and post
answers with the null hypothesis that the medians of the pre and post-test are similar. The null
hypothesis was rejected in all cases with p<0.05. Students display a statistically significant increase in familiarity with vocabulary items related to
Gravitational Wave (GW) production and detection principles (22% increase; p<0.05) as well as
familiarity with items related to technical aspects of GW measurement (15.7% increase; p<0.05). Furthermore, students’ answers to an open-ended question regarding how they would explain
Gravitational Waves to a friend demonstrate a significant increase in their fluency (29.7% increase of
scientifically correct answers; p<0.05). The findings of the present study indicate that students demonstrate a statistically significant
increase in confidence to explain Virgo to a friend (33.3% increase; p<0.05). The findings of the present study indicate that students demonstrate a statistically significant
increase in confidence to explain Virgo to a friend (33.3% increase; p<0.05). Students displayed an increase in their understanding of wave interference and interferometry (77%
increase; p<0.05), something that can be explained taking into account that the visit’s main theme
was the detection of gravitational waves, the principle of which is based on the phenomenon of
wave interference. As this phenomenon was discussed in detail in the introductory presentation, the
visit to the Michelson interferometer exhibit as well as the visit to the Virgo interferometer itself, it
is understood that students’ understanding of destructive interference of light is improved,
however, not adequately taking into account that less than 50% of the students answered correctly
in the pre and post questionnaire respectively. It is proposed that students who attend the visit do
prior preparation in the classroom regarding the phenomenon of wave interference. Figure 10: Screenshot from virtual visit to Virgo 617 students from 29 schools joined the visit,
out of which a subsample of N=101 students
from 24 schools offered matching pre – and
post- questionnaires (response rate of16.8%). Out of the N=101 students, 50 were female and
47 male, with 4 not specifying. The majority of
the respondents were high school students
with age distribution: [12,15): 9 students;
[15,19): 68 students; [19,20]: 20 students, while
4 students didn’t answer the question. Regarding the students’ nationalities: 46 students came from Greece, 25 from Italy, 13 from
Pakistan, 12 from Romania, 3 from Portugal, 1 from Austria and 1 from France. The students were
Figure 10: Screenshot from virtual visit to Virgo 58 Masterclasses A series of Masterclasses were organised by project partners IASA with school students and their
teachers being given the opportunity to learn about Nobel prize winning physics. Facilitated by the 59 59 59 59 Universities of Athens and Crete, students were given a hands-on experience of using the HYPATIA
event display to look for Z and Higgs bosons, as used in the FRONTIERS demonstrators. Teachers
were introduced to the FRONTIERS Project with the Demonstrators presented. Over 8 Masterclass events, held both in-person
and virtually, 405 students and 71 teachers took
part. These events further supported and
promoted the engagement of teachers and
students in the FRONTIERS Demonstrators and the
meaningful engagement with frontier physics. Figure 11: A screenshot from an online Masterclass
event. Universities of Athens and Crete, students were given a hands-on experience of using the HYPATIA
event display to look for Z and Higgs bosons, as used in the FRONTIERS demonstrators. Teachers
were introduced to the FRONTIERS Project with the Demonstrators presented. events, held both in-person
udents and 71 teachers took
urther supported and
ement of teachers and
NTIERS Demonstrators and the
ment with frontier physics. Figure 11: A screenshot from an online Masterclass
event. Over 8 Masterclass events, held both in-person
and virtually, 405 students and 71 teachers took
part. These events further supported and
promoted the engagement of teachers and
students in the FRONTIERS Demonstrators and the
meaningful engagement with frontier physics. Figure 11: A screenshot from an online Masterclass
event. 60 60 Community Support Environment A central aim of the FRONTIERS project was to create a sustainable and expanding network of
teachers to collaborate with researchers and students to introduce Nobel Prize Physics in education. This was provided through the FRONTIERS Community Support Environment. One of the main messages coming from teachers who have participated in outreach activities
regarding frontier science is that they need systematic and professional support to be able to
effectively teach these issues to their students. This means that they need to feel part of a
community of practice, and to have regular interaction with experts. A forum for the exchange of
ideas and practices with their peers was seen to facilitate teachers feeling confident enough to teach
complex scientific issues in an understandable and systematic fashion. In the FRONTIERS community support environment, the participants can discuss with their peers and
interact with researchers in the fields of frontier Physics and Science Education in a systematic
fashion. The community environment also allows members to explore the project’s resources,
participate in debates, share projects, upload their own resources, and participate in online activities
such as webinars. Furthermore, members of the FRONTIERS online community can receive
continuous support by the project partners. The desired characteristics of the FRONTIERS Community were: The desired characteristics of the FRONTIERS Community were: a. Provide opportunities for sustainable engagement and networking: -
Systematic interaction with experts in the fields of frontier physics. - Collaboration with peers sharing the same vision and facing the same challenges, from all over
the world. b. Offer continuous professional support: -
Provide teachers with training materials and model educational activities for their classroom. -
Organize participatory engagement activities for teachers’ professional development. -
Encourage teachers to become content creators. c. Offer appreciation and recognition -
Offer teachers tangible appreciation and recognition for their achievements. -
Encourages teacher discussion and pay attention to their needs and concerns. -
Boost the self-confidence of teachers in teaching frontier physics. -
Offer teachers an active role and give them opportunities to increase their status. -
Offer a platform for text-based and multimedia interaction between community members. -
Offer a platform for text-based and multimedia interaction between community members. -
Offer a repository of unique educational resources, scientific and educational tools as well as
authoring tools for content creation. -
Offer a platform for the organization of virtual participatory engagement and training activities. Facebook Facebook
The Facebook platform has been utilised by
the FRONTIERS Project to engage with physics
teachers and the wider public. The platform has been used to share
FRONTIERS project news, upcoming events
and live streaming events such as virtual visits
to research facilities. It has also been used to
share teacher and student generated material
adapted from the FRONTIERS Demonstrators,
as well as other physics related content. The FRONTIERS Facebook page has a total
reach of 155,129 Facebook users. The page has 1,111 likes, with 1,227 people actively following the
page. Of the Facebook users that like the FRONTIERS page, 63% are female and the largest cohort
aged between 45 and 54 years. The FRONTIERS project Facebook page has a broad international reach. Of the Facebook users that
liked the Frontiers page, most users came from Greece, Portugal and Italy. Also in the top ten
countries that users came from included Turkey, Bulgaria, the United Kingdom and India. Figure 12: The FRONTIERS Project Facebook page s
s
al
,
as 1,111 likes, with 1,227 people actively following the
ONTIERS page, 63% are female and the largest cohort
Figure 12: The FRONTIERS Project Facebook page Figure 12: The FRONTIERS Project Facebook page The Facebook platform has been utilised by
the FRONTIERS Project to engage with physics
teachers and the wider public. The platform has been used to share
FRONTIERS project news, upcoming events
and live streaming events such as virtual visits
to research facilities. It has also been used to
share teacher and student generated material
adapted from the FRONTIERS Demonstrators,
as well as other physics related content. as well as other physics related content. The FRONTIERS Facebook page has a total
reach of 155,129 Facebook users. The page has 1,111 likes, with 1,227 people actively following the
page. Of the Facebook users that like the FRONTIERS page, 63% are female and the largest cohort
aged between 45 and 54 years. The FRONTIERS project Facebook page has a broad international reach. Of the Facebook users that
liked the Frontiers page, most users came from Greece, Portugal and Italy. Also in the top ten
countries that users came from included Turkey, Bulgaria, the United Kingdom and India. Figure 12: The FRONTIERS Project Facebook page Figure 12: The FRONTIERS Project Facebook page The FRONTIERS Facebook page has a total
reach of 155,129 Facebook users. Community Support Environment -
Offer a platform for the organization of virtual participatory engagement and training activities. To develop and facilitate the FRONTIERS Community and realise the above-mentioned desired
characteristics, the following online FRONTIERS Project resources have been utilised. To develop and facilitate the FRONTIERS Community and realise the above-mentioned desired
characteristics, the following online FRONTIERS Project resources have been utilised. 1. Facebook
2. Website
3. Open Schools for Open Societies Platform
4. Summer and Winter Schools
5. Google Classroom 61
Each of the FRONTIERS Project online resources will be examined and evaluated as contributing
towards the FRONTIERS Project Community. Each of the FRONTIERS Project online resources will be examined and evaluated as contributing
towards the FRONTIERS Project Community. 61 Facebook The page has 1,111 likes, with 1,227 people actively following the
page. Of the Facebook users that like the FRONTIERS page, 63% are female and the largest cohort
aged between 45 and 54 years. The FRONTIERS project Facebook page has a broad international reach. Of the Facebook users that
liked the Frontiers page, most users came from Greece, Portugal and Italy. Also in the top ten
countries that users came from included Turkey, Bulgaria, the United Kingdom and India. The FRONTIERS project Facebook page has a broad international reach. Of the Facebook users that
liked the Frontiers page, most users came from Greece, Portugal and Italy. Also in the top ten
countries that users came from included Turkey, Bulgaria, the United Kingdom and India. Website
FRONTIERS Project website has been
developed as an online presence for all
relevant information and updates
relating to the project. This includes
details of project partners, outputs and
community. The website hosts regular news updates
as well as access to the developed Nobel
prize winning physics demonstrators. Figure 13: The FRONTIERS website homepage Also hosted on the website are best-
practice examples of developed resources
from the FRONTIERS Community Teachers
for other teachers to engage with and use
in their own classrooms. Figure 14: The FRONTIERS Demonstrators hosted on the project
website The FRONTIERS website had 8,305 visitors
between January 2019 and August 2021. These users accounted for 12,305 sessions
on the websites, giving an average of 1.48
visits per user. During these sessions each
user visited an average of 2.15 pages on the
website. Figure 14: The FRONTIERS Demonstrators hosted on the project
website Similar to the FRONTIERS Facebook page, the project website had a broad international audience. The top 10 preferred languages of the website users included English, Greek and Arabic. Similar to the FRONTIERS Facebook page, the project website had a broad international aud
The top 10 preferred languages of the website users included English, Greek and Arabic. he top 10 preferred languages of the website users included English, Greek and Arabic. 0
500
1000
1500
2000
2500
English - US
Greek
Arabic
English - UK
Portuguese
French
Arabic - EG
Italian
Arabic - AE
French
Top Ten Language Preferences of Website
Users Website FRONTIERS Project website has been
developed as an online presence for all
relevant information and updates
relating to the project. This includes
details of project partners, outputs and
community. The website hosts regular news updates
as well as access to the developed Nobel
prize winning physics demonstrators. Figure 13: The FRONTIERS website homepage 62 62 Open Schools for Open Societies The FRONTIERS Community aimed to
encourage the cooperation between teachers,
students and researchers and create an online
experience that engages educators in sharing
their best science teaching practices. Figure 15: The Frontiers Demonstrators hosted on OSOS
platform Following detailed comparative research of the
available online platforms, the project
consortium chose Open Schools for Open
Societies (OSOS) platform to host the project
Demonstrators and other community building
activities. The OSOS portal is the online
educational portal for Open Schooling in Figure 15: The Frontiers Demonstrators hosted on OSOS
platform 63 Europe, hosting 100 vibrant online communities of 1000 schools and engaging more than 1500
teachers. Europe, hosting 100 vibrant online communities of 1000 schools and engaging more than 1500
teachers. The OSOS platform provided the opportunity to create educational resources using a dedicated
authoring environment, the ISE authoring tool. This tool facilitated the hosting of the FRONTIERS
Demonstrators following the inquiry based pedagogical approach adopted by the project partners. The OSOS platform provided the opportunity to create educational resources using a dedicated
authoring environment, the ISE authoring tool. This tool facilitated the hosting of the FRONTIERS
Demonstrators following the inquiry based pedagogical approach adopted by the project partners. Educational scenarios using the ISE authoring tool
allowed partners to embed multimedia (videos,
photos) as well as virtual labs and datasets in an
educational scenario. This allowed FRONTIERS
teachers and their students to engage with the
Demonstrators and the Nobel prize winning physics
behind them in a more meaningful and real way. Figure 16: The inquiry-based pedagogical
framework support by the OSOS platform Figure 16: The inquiry-based pedagogical
framework support by the OSOS platform Embedding the multimedia resources and virtual lab
facilities with the Demonstrators on the OSOS
platform, facilitated the teacher and student to
undertake real-world scientific work using the tools Figure 16: The inquiry-based pedagogical
framework support by the OSOS platform All 21 FRONTIERS Demonstrators and associated resources are hosted on the OSOS platform. All 21 FRONTIERS Demonstrators and associated resources are hosted on the OSOS platfor The OSOS platform also hosts communities of educators in each of the partner countries with
resources in their native language. There are six communities hosted on the OSOS; International,
Greek, Irish, French, Italian and Portuguese. There are 174 active members across these virtual
communities. International e-Schools In addition to the initial training of teachers across the project partner countries, three online
International e-Schools have taken place. The training provided to these FRONTIERS Community
members was designed and implemented systematically upon the finalisation of the project’s
demonstrators. Training activities offered teachers an in-depth view of the project’s pedagogical framework;
proposed educational activities helping teachers to enhance their content knowledge. Furthermore,
connections from FRONTIERS resources with the school curricula were highlighted and developed. Teachers were given the opportunity to take part in virtual visits to research facilities and
encouraged to bring them to the classroom. Finally, the training activities aimed to empower
teachers to adapt and create their own content. Participating teachers to the International e-Schools were able to interact synchronously or
asynchronously with the content of the training, interact with their trainers and peers, and complete
given tasks provided by the FRONTIERS community environment. 277 participants have fully completed the International e-Schools so far, with many others joining for
virtual visits hosted as part of the e-School events. This group of teachers who have been engaged,
trained and supported in integrating Nobel prize winning physics into their classes, is evidence of the
active FRONTIERS Community of motivated teachers of physics across the EU and beyond. 64 64 Google Classroom Google Classroom
Google Classroom offers teachers the
potential for organising focus groups
online, uploading exercises, receiving
students’ answers and initiating dedicated
discussions. There are 70 teachers actively enrolled in
the FRONTIERS Google Classroom. An international google classroom has
been developed alongside each national
Frontiers Google classroom. These classrooms are used to engage with FRONTIERS Teachers who
join, communicating FRONTIERS Project updates, upcoming training events, relevant other
materials, as well as an opportunity for the community to engage and support each other. Figure 17: The FRONTIERS Project on Google Classroom ers the
groups
ceiving
ng dedicated
enrolled in
om. oom has
h national
hese classrooms are used to engage with FRONTIERS Teachers who
RS Project updates, upcoming training events, relevant other
unity for the community to engage and support each other. Figure 17: The FRONTIERS Project on Google Classroom Figure 17: The FRONTIERS Project on Google Classroom Google Classroom offers teachers the
potential for organising focus groups
online, uploading exercises, receiving
students’ answers and initiating dedicated
discussions. There are 70 teachers actively enrolled in
the FRONTIERS Google Classroom. An international google classroom has
been developed alongside each national
Frontiers Google classroom. These classrooms are used to engage with FRONTIERS Teachers who
join, communicating FRONTIERS Project updates, upcoming training events, relevant other
materials, as well as an opportunity for the community to engage and support each other. Figure 17: The FRONTIERS Project on Google Classroom An international google classroom has
been developed alongside each national Frontiers Google classroom. These classrooms are used to engage with FRONTIERS Teachers who
join, communicating FRONTIERS Project updates, upcoming training events, relevant other
materials, as well as an opportunity for the community to engage and support each other. 65 65 References Bamberger, M., Vijayendra, R., & Woolcock, M. (2010). Using mixed methods in monitoring and
evaluation. In Sage Handbook of mixed methods in social and behavioural research (2nd ed., pp. 613–641). SAGE. Banchi, H., & Bell, R. (2008). The many levels of inquiry: Inquiry comes in various forms. Science and
Children, 46(2), 26–29. Creswell, J. W., & Plano Clark, V. L. (2018). Designing and conducting mixed methods research (3rd
edition). Sage. Johnson, R. B., Onwuegbuzie, A. J., & Turner, L. A. (2007). Toward a Definition of Mixed Methods
Research. Journal of Mixed Methods Research, 1(2), 112–133. https://doi.org/10.1177/1558689806298224 Meyer, D. Z., Antink Meyer, A., Nabb, K. A., Connell, M. G., & Avery, L. M. (2013). Google Classroom A Theoretical and
Empirical Exploration of Intrinsic Problems in Designing Inquiry Activities. Research in Science
Education, 43(1), 57–76. https://doi.org/10.1007/s11165-011-9243-4 National Research Council. (1996). National Science Education Standards. National Academies Press. https://www.nap.edu/catalog/4962 Onwuegbuzie, A. J., & Hitchcock, J. H. (2017). A meta-framework for conducting mixed methods
impact evaluations: Implications for altering practice and the teaching of evaluation. Studies in
Educational Evaluation, 53, 55–68. https://doi.org/10.1016/j.stueduc.2017.02.001 Streich, I., & Mayer, J. (2020). Effects and Prerequisites of Self-Generation in Inquiry-Based Learning. Education Sciences, 10(10), 1. https://doi.org/10.3390/educsci10100277 Streich, I., & Mayer, J. (2020). Effects and Prerequisites of Self-Generation in Inquiry-Based Learning. Education Sciences, 10(10), 1. https://doi.org/10.3390/educsci10100277 66 66
|
https://openalex.org/W2166272159
|
https://figshare.com/articles/journal_contribution/Long-term_5_year_safety_of_bronchial_thermoplasty_Asthma_Intervention_Research_AIR_trial/10149833/1/files/18291974.pdf
|
English
| null |
Long-term (5 year) safety of bronchial thermoplasty: Asthma Intervention Research (AIR) trial
|
BMC pulmonary medicine
| 2,011
|
cc-by
| 8,248
|
© 2011 Thomson et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative
Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and
reproduction in any medium, provided the original work is properly cited. Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Long-term (5 year) safety of bronchial
thermoplasty: Asthma Intervention
Research (AIR) trial Neil C Thomson1*, Adalberto S Rubin2, Robert M Niven3, Paul A Corris4, Hans Christian Siersted5,
Ronald Olivenstein6, Ian D Pavord7, David McCormack8, Michel Laviolette9, Narinder S Shargill10, Gerard Cox11,
the AIR Trial Study Group Abstract Background: Bronchial thermoplasty (BT) is a bronchoscopic procedure that improves asthma control by reducing
excess airway smooth muscle. Treated patients have been followed out to 5 years to evaluate long-term safety of
this procedure. Methods: Patients enrolled in the Asthma Intervention Research Trial were on inhaled corticosteroids ≥200 μg
beclomethasone or equivalent + long-acting-beta2-agonists and demonstrated worsening of asthma on long-
acting-b2-agonist withdrawal. Following initial evaluation at 1 year, subjects were invited to participate in a 4 year
safety study. Adverse events (AEs) and spirometry data were used to assess long-term safety out to 5 years post-BT. Results: 45 of 52 treated and 24 of 49 control group subjects participated in long-term follow-up of 5 years and
3 years respectively. The rate of respiratory adverse events (AEs/subject) was stable in years 2 to 5 following BT (1.2,
1.3, 1.2, and 1.1, respectively,). There was no increase in hospitalizations or emergency room visits for respiratory
symptoms in Years 2, 3, 4, and 5 compared to Year 1. The FVC and FEV1 values showed no deterioration over the
5 year period in the BT group. Similar results were obtained for the Control group. Conclusions: The absence of clinical complications (based on AE reporting) and the maintenance of stable lung
function (no deterioration of FVC and FEV1) over a 5-year period post-BT in this group of patients with moderate
to severe asthma support the long-term safety of the procedure out to 5 years. * Correspondence: neil.thomson@glasgow.ac.uk
1Gartnavel General Hospital, University of Glasgow, Glasgow, UK
Full list of author information is available at the end of the article Study subjects All patients completing the 12-month follow-up evalua-
tions in the AIR Trial (BT group: standard-of-care +
BT; Control group: standard-of-care) were invited to
participate in a 5 year post-treatment extension study to
evaluate longer-term safety under a new protocol
(NCT00448812). The exclusion criteria for participation
in the extension study were: participation in another
clinical trial involving respiratory intervention, or new
diagnosis of psychiatric disorder which in the judgment
of the investigator could interfere with provision of
informed consent, completion of tests, therapy, or
follow-up. During Year 1, adverse events were solicited during 12
office visits and 9 telephone contacts over the course of
the year, as well as a review of the medical chart. During
the longer-term follow-up, adverse events were actively
solicited from the subject during the annual evaluation
and through a review of the medical chart for the prior
year for subjects managed at the investigator’s institute. The recording of adverse events in Year 1 (AIR Trial)
differed from the subsequent years (AIR Extension
Study) in that in Year 1, multiple symptoms associated
with an adverse event were collected as individual
adverse events; while in the subsequent years, an adverse
event with multiple symptoms was counted as a single
adverse event e.g., multiple respiratory symptoms asso-
ciated with worsening of asthma were considered as a
single event called “asthma (multiple symptoms)”
adverse event. The key inclusion criteria to establish eligibility of
patients to participate in the original AIR Trial were:
18-65 year old ambulatory adults; stable asthma (no
unscheduled visits or change to medications within
6 weeks prior to randomization); requiring inhaled corti-
costeroids (ICS): at least 200 μg beclomethasone or
equivalent per day, and long-acting b2-agonist (LABA):
at least 100 μg salmeterol or equivalent per day; demon-
stration of worsening of asthma after 2-week LABA
withdrawal; pre-bronchodilator FEV1 ≥60% and ≤85%
predicted; methacholine PC20 < 8 mg/ml; and, non-
smoker x 1 yr; if former smoker, less than 10 pack-year
history. The key exclusion criteria were: history of ≥3
lower respiratory tract infections per year (requiring
antibiotics); and, requirement of > 4 puffs/day of a
short-acting b2-agonist, excluding for exercise. The pro-
tocol for the longer-term extension study was approved
by the respective Institutional Review Boards/Ethics
Committees at each participating institution prior to Study procedures Subjects in the BT group were evaluated annually; at
Year 2, Year 3, Year 4, and Year 5 after their last treat-
ment bronchoscopy. Subjects in the Control group were
evaluated at Year 2 and Year 3 and then exited from the
study. Year 1 data are provided for matched pairs in
both the BT and Control groups comprising those sub-
jects that enrolled in the longer-term follow-up. Annual evaluations included a physical examination,
pre- and post-bronchodilator spirometry, static lung
volumes, diffusing capacity, chest x-ray (PA and Lateral),
methacholine PC20 (out to Year 3 only), as well as active
solicitation of information on any adverse events, emer-
gency room visits and hospitalizations for asthma symp-
toms, oral corticosteroid pulses for worsening asthma
symptoms, and any changes in maintenance asthma
medications. Maintenance asthma medication use, oral
corticosteroid use, adverse events, emergency room vis-
its and hospitalizations were verified through medical
record review for about 80% of the subjects whose pri-
mary care was under the supervision of the investigator
at their respective institution. X-ray observations
reported by radiologists at each site were collectively
reviewed by an independent pulmonologist to assess
clinical relevance of the observations if any. Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Page 2 of 9 obtaining a signed informed consent from the study
participants. the percentage of symptom-free days and symptom
scores, while fewer puffs of rescue medication were
required [9]. Two additional randomized controlled
trials, the Research in Severe Asthma (RISA) Trial [10],
and the sham-controlled Asthma Intervention Research
2 (AIR2) Trial [11], have provided additional support for
the effectiveness of BT. The safety of BT over the post-
treatment period out to one year was established in all 3
randomized clinical studies. Because BT is a novel treat-
ment that alters the amount of airway smooth muscle,
and asthma is a chronic disease that is associated with
structural changes in the airway wall, it is important to
know whether this treatment is associated with any
longer term adverse outcomes. Longer-term safety data
out to 5 years are available for a small cohort of BT
treated subjects from the first clinical study of patients
with mild to moderate asthma [12,13]. We now describe
the safety profile of BT out to 5 years post-treatment
from patients with moderate to severe asthma from the
AIR Trial [9]. the percentage of symptom-free days and symptom
scores, while fewer puffs of rescue medication were
required [9]. Two additional randomized controlled
trials, the Research in Severe Asthma (RISA) Trial [10],
and the sham-controlled Asthma Intervention Research
2 (AIR2) Trial [11], have provided additional support for
the effectiveness of BT. The safety of BT over the post-
treatment period out to one year was established in all 3
randomized clinical studies. Because BT is a novel treat-
ment that alters the amount of airway smooth muscle,
and asthma is a chronic disease that is associated with
structural changes in the airway wall, it is important to
know whether this treatment is associated with any
longer term adverse outcomes. Longer-term safety data
out to 5 years are available for a small cohort of BT
treated subjects from the first clinical study of patients
with mild to moderate asthma [12,13]. We now describe
the safety profile of BT out to 5 years post-treatment
from patients with moderate to severe asthma from the
AIR Trial [9]. Background provides an additional option for managing patients
with severe asthma. BT provides therapeutic benefit by
reducing the amount of excess smooth muscle in the
airways, with the resultant effect of reducing broncho-
constriction in response to asthma triggers. Results from
the Asthma Intervention Research (AIR) Trial, the first
randomized clinical trial of BT which compared BT plus
standard-of-care therapy (inhaled corticosteroids (ICS)
and long-acting-b2-agonists (LABA)) to standard-of-care
alone, demonstrated that the mean rate of mild exacer-
bations, as compared with baseline, was reduced in the
BT group but was unchanged in the control group. Furthermore at 12 months, when subjects were on ICS
alone, there were significantly greater improvements in
the BT group than in the control group in the morning
peak expiratory flow, scores on the AQLQ and ACQ, Asthma continues to be a major health concern world-
wide, with over 23 million people in the United States
who suffer with this disease [1]. Approximately 5-10% of
these patients are characterized as having severe persis-
tent asthma based on continued presence of asthma
symptoms despite treatment with current state-of-the-
art medications [2]. Poorly controlled asthma impacts
the patient’s quality of life, increases healthcare utiliza-
tion, and imposes both a social as well as an economic
burden [3-8]. The recent approval of the Alair® Bronchial Thermo-
plasty System for delivering bronchial thermoplasty (BT) * Correspondence: neil.thomson@glasgow.ac.uk
1Gartnavel General Hospital, University of Glasgow, Glasgow, UK
Full list of author information is available at the end of the article Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Adverse Events There were no incidences of pneumothorax, intubation,
mechanical ventilation, cardiac arrhythmias, or death as
a result of BT treatment over the 5 year follow-up. Table 1 Number of Subjects Evaluated Annually out to
5 Years
Subjects
Completing 1
Year Follow-up
in AIR Trial
Subjects
Enrolling for
Longer-Term
Follow-up
Subjects Completing
Follow-up
Year
2
Year
3
Year
4
Year
5
BT
52
45
41a
41b
43
42c
Control
49
24
23d
21e
-
-
a: 4 subjects missed visit. b: 2 subjects missed visit; 1 subject withdrew consent; 1 subject Lost-to-
Follow-up. c: 1 subject missed visit. d: 1 subject missed visit. e: 2 subjects withdrew consent after completing Year 2 evaluation, and 1
subject withdrew consent mid-Year 3. Control group subjects exited from study after completing Year 3 evaluation. Table 1 Number of Subjects Evaluated Annually out to
5 Years Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Page 3 of 9 Respiratory adverse events reported during the course
of the 5 year follow-up are summarized in Table 3. Dur-
ing Year 1, the rate of respiratory adverse events in both
the BT and Control groups was higher as a result of the
method of recording the adverse events whereby multi-
ple symptoms associated with an adverse event were
recorded as separate adverse events. In subsequent years
(Year 2 to Year 5), an adverse event with multiple symp-
toms was recorded as a single adverse event. The rate of
respiratory adverse events in the BT group (AEs/subject)
remained stable in years 2 to 5 following BT. A repeated
measures analysis for the “asthma (multiple symptoms)”
adverse events which reflects worsening asthma control
showed no deterioration over time from Year 2 to Year
5 (p = 0.47). During Year 2 and Year 3, when data for
the Control group was collected, the respiratory adverse
event rate between the BT and Control groups was not
significantly different. Fisher’s Exact test was used to compare proportion of
subjects with respiratory hospitalizations and emergency
room visits in the BT and Control groups during Years
1, 2 and 3. Trends in the percent of subjects with hospi-
talizations or emergency room visits for respiratory
symptoms across Years 1 to 5 were investigated using a
repeated measures logistic regression (generalized esti-
mating equation), modeling the percent of subjects
reporting the event. Maintenance medications ICS dose:
Change from Baseline to each follow-up year in ICS
dose was analyzed with a Signed Rank test. Statistical Analyses
Demographics Group means were compared using Student’s t-test. Hospitalizations and Emergency Room Visits for respira-
tory symptoms: The respective number of subjects com-
pleting each annual follow-up visit was used to calculate
the proportion of subjects with hospitalizations or emer-
gency room visits for respiratory symptoms in each year. Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Results Demographics and Clinical Characteristics
Forty five (45) of the 52 subjects in the BT group (87%),
and 24 of the 49 subjects in the Control group (49%)
who completed the Year 1 evaluation opted to partici-
pate in the extension study. The 7 subjects in the BT
group and 25 subjects in the Control group who
declined to participate in the long-term follow-up did so
for personal reasons, and not due to mortality. The
numbers of subjects enrolling for the longer-term fol-
low-up and those completing scheduled annual evalua-
tions are summarized in Table 1. g
y
Respiratory adverse events occurring at a by-subject
incidence rate of ≥3.0% in any of the years are given in
Table 4. For the majority of respiratory adverse events,
the incidence rates were stable during each year from
Year 2 to Year 5 in the BT group, and from Year 2 and
Year 3 in the Control group. The respiratory adverse
events were typical of asthma. One subject in the BT
group who had undergone the procedure was diagnosed
with a lung abscess in the previously treated left upper
lobe at 14 months (Year 2), and was resolved with surgi-
cal resection. The subject had undergone BT unevent-
fully and had completed the 12 month follow-up with
normal spirometric values and good asthma control
(Post-BD FEV1 at baseline was 2.27L, and at 12 months
was 2.33L). Histological examination of the dissected
lung did not reveal an obstruction or any other poten-
tially contributory abnormality in the airways as a result
of the treatment. The abscess was considered secondary
to an infection. At the time of exit from the study at
5 years, the post-BD FEV1 for this subject was 1.78L
compared to baseline value of 2.27L. The baseline demographic information and clinical
characteristics (at time of entry into the AIR Trial) for
subjects participating in the extension study are pro-
vided in Table 2. The groups were well matched with
no statistically significant differences between them for
any given parameter. Healthcare Utilization Events Subjects)
3 (3)
2 (2)
Lung Function Measures
Morning PEF (L/min)
368.4 ± 99.7
394.1 ± 111.7
Pre-Bronchodilator FEV1 (% predicted)
72.5 ± 10.9
74.9 ± 8.9
Post-Bronchodilator FEV1 (% predicted)
84.4 ± 13.8
86.1 ± 9.5
Diffusion Capacity (mL/min/mm Hg)
15.7 ± 10.7
15.9 ± 11.7
Total Lung Capacity (L)
6.0 ± 1.2
5.9 ± 1.3
Residual Volume (L)
2.1 ± 0.7
2.0 ± 0.7
Methacholine PC20 (mg/ml)
0.25
0.28
Geometric mean (range)
(0.2, 0.4)
(0.1, 0.6)
Definition of abbreviations: BT = Bronchial Thermoplasty; LABA = Long-Acting b2-Agonist;
PEF = Peak Expiratory Flow Rate; FEV1 = Forced Expiratory Volume in 1 second. Table 2 Baseline Demographics and Clinical Characteristics c: Patient reported. BT versus Control: All parameters, Not significant (Student’s t-test of the mean). in Years 2, 3, 4, and 5 compared to Year 1 (Table 5)
(p = 0.55; repeated measures analysis). The number of
ER visits for respiratory symptoms in the Control group
in Years 2 and 3 were comparable to the BT group (p =
0.41 and p = 1.00, respectively; Fisher’s Exact test). Oral Corticosteroid Use for Asthma Symptoms in Years 2, 3, 4, and 5 compared to Year 1 (Table 5)
(p = 0.55; repeated measures analysis). The number of
ER visits for respiratory symptoms in the Control group
in Years 2 and 3 were comparable to the BT group (p =
0.41 and p = 1.00, respectively; Fisher’s Exact test). Oral Corticosteroid Use for Asthma Symptoms The frequency of OCS usage for worsening of asthma symp-
toms is shown in Table 6. Neither the rate of OCS usage nor
the proportion of subjects requiring OCS pulses showed any
worsening over the 5 year period in the BT group. OCS The frequency of OCS usage for worsening of asthma symp-
toms is shown in Table 6. Neither the rate of OCS usage nor
the proportion of subjects requiring OCS pulses showed any
worsening over the 5 year period in the BT group. Healthcare Utilization Events During Year 1 and Year 2, more subjects in the BT
group required hospitalizations for respiratory symp-
toms than the Control group, but these differences were
not significant. There was one hospitalization for
respiratory symptoms in the Control group in Year 3. Over the course of the 5 year post-BT follow-up, the
number of hospitalizations, and the proportion of sub-
jects experiencing hospitalizations for respiratory symp-
toms did not get worse compared to Year 1 after BT
(Table 5) (p = 0.16; repeated measures analysis for pro-
portion of subjects). Similarly, the number of emergency
room (ER) visits for respiratory symptoms and the pro-
portion of subjects experiencing ER visits for respiratory
symptoms remained comparable and did not get worse Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Page 4 of 9 Page 4 of 9 Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Table 2 Baseline Demographics and Clinical Characteristics
BT (n = 45)
Control (n = 24)
Age (yrs)
40.0 ± 11.2
40.8 ± 12.1
Gender
Male 19 (42%)
Male 9 (38%)
Female 26 (58%)
Female 15 (63%)
Race
White 41 (91%)
White 22 (92%)
Black 3 (7%)
Black 2 (8%)
Asian 1 (2%)
Asian 0 (0%)
Height (cm)
166.1 ± 9.6
164.8 ± 7.7
Weight (kg)
76.3 ± 23.3
77.7 ± 16.9
Inhaled Corticosteroid Dose (μg)a
1305 ± 880
1141 ± 1053
LABA Dose (μg)b
109 ± 34
100 ± 15
Symptom-Free Days (%)
33.3 ± 34.3
46.1 ± 41.0
Asthma Control Questionnaire (ACQ) Score
1.3 ± 0.6
1.2 ± 0.7
Asthma Quality of Life Questionnaire (AQLQ) Score
5.6 ± 0.9
5.6 ± 0.9
Rescue Medication Use (No. of puffs/7days)
10.6 ± 14.7
5.5 ± 10.4
Emergency Room Visits for Respiratory Symptoms in prior 12 monthsc
No. Events (No. Subjects)
3 (3)
0 (0)
Hospitalizations for Respiratory Symptoms in prior 12 monthsc
No. Events (No. Healthcare Utilization Events OCS Table 3 Summary of Respiratory Adverse Events
Year 1
Year 2
Year 3
Year 4
Year 5
Number of subjects
BT
45a
45
43
43
42
Control
24a
24
21
– b
– b
Number of subjects reporting (Percent of subjects)
BT
38 (84%)
24 (53%)
24 (56%)
23 (53%)
22 (52%)
Control
18 (75%)
13 (54%)
12 (57%)
–
–
Events per subject
BT
4.5
1.2
1.3
1.2
1.1
Control
3.1
1.2
1.3
–
–
a: Year 1 data only for subjects who enrolled for longer-term follow-up. b: Control group subjects exited from Study after Year 3 evaluations. Table 3 Summary of Respiratory Adverse Events Thomson et al. Table 3 Summary of Respiratory Adverse Events BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Page 5 of 9 Page 5 of 9 Table 4 Subjects with Respiratory Adverse Events (All events reported at ≥3.0% in any year)
Year 1a
Year 2
Year 3
Year 4b
Year 5b
Adverse Event
BT
(n = 45)
Control
(n = 24)
BT
(n = 45)
Control
(n = 24)
BT
(n = 43)
Control
(n = 21)
BT
(n = 43)
BT
(n = 42)
Dyspnea
19 (42.2%)
12 (50.0%)
4 (8.9%)
3 (12.5%)
4 (9.3%)
3 (14.3%)
4 (9.3%)
4 (9.5%)
Cough
17 (37.8%)
7 (29.2%)
4 (8.9%)
1 (4.2%)
2 (4.7%)
3 (14.3%)
3 (7.0%)
2 (4.8%)
Wheeze
14 (31.1%)
4 (16.7%)
2 (4.4%)
1 (4.2%)
3 (7.0%)
1 (4.8%)
3 (7.0%)
2 (4.8%)
Nasal congestion
13 (28.9%)
5 (20.8%)
2 (4.4%)
0
0
0
0
1 (2.4%)
Upper Respiratory Tract Infection
10 (22.2%)
2 (8.3%)
11 (24.4%)
4 (16.7%)
8 (18.6%)
4 (19.1%)
8 (18.6%)
4 (9.5%)
Productive cough
9 (20.0%)
5 (20.8%)
2 (4.4%)
1 (4.2%)
2 (4.7%)
0
0
1 (2.4%)
Chest discomfort
8 (17.8%)
3 (12.5%)
2 (4.4%)
2 (8.3%)
3 (7.0%)
1 (4.8%)
1 (2.3%)
2 (4.8%)
Nasopharyngitis
6 (13.3%)
0
1 (2.2%)
0
0
0
1 (2.3%)
1 (2.4%)
Nocturnal Dyspnea
6 (13.3%)
2 (8.3%)
0
0
0
0
0
0
Respiratory Tract Infectionc
5 (11.1%)
5 (20.8%)
3 (6.7%)
2 (8.3%)
5 (11.6%)
1 (4.8%)
5 (11.6%)
4 (9.5%)
Pharyngolaryngeal pain
5 (11.1%)
3 (12.5%)
0
0
0
0
0
0
Respiratory Tract congestion
4 (8.9%)
2 (8.3%)
0
0
0
0
0
0
Discolored sputum
4 (8.9%)
0
3 (6.7%)
0
0
0
0
0
Rhinitis
2 (4.4%)
0
0
0
1 (2.3%)
0
0
2 (4.8%)
Bronchitisd
1 (2.2%)
0
1 (2.2%)
1 (4.2%)
1 (2.3%)
2 (9.5%)
1 (2.3%)
1 (2.4%)
Pharyngitis
1 (2.2%)
1 (4.2%)
0
0
0
0
0
0
Pleuritic Pain
1 (2.2%)
1 (4.2%)
0
0
0
0
0
0
Rhinorrhea
1 (2.2%)
1 (4.2%)
0
0
1 (2.3%)
0
0
0
Asthma (multiple symptoms)e
0
0
4 (8.9%)
2 (8.3%)
7 (16.3%)
1 (4.8%)
7 (16.3%)
6 (14.3%)
Sinusitis
0
0
1 (2.2%)
1 (4.2%)
2 (4.7%)
0
2 (4.7%)
2 (4.8%)
Nasal polyps
0
0
1 (2.2%)
0
0
0
2 (4.7%)
0
Pneumonia
0
0
0
0
1 (2.3%)
1 (4.8%)
0
0
a: Year 1 data only for subjects who enrolled for longer-term follow-up. Table 3 Summary of Respiratory Adverse Events Adverse events solicited from patient during multiple office visits in Year 1. In
subsequent years, adverse events solicited only at annual follow-up visit. b: Control group subjects exited from Study after Year 3 evaluations. c: Includes adverse events reported as “Respiratory Tract Infection” and “Lower Respiratory Tract Infection”. d: Includes adverse events reported as “Bronchitis” and “Tracheobronchitis”. e: Asthma - In Year 1, all symptoms were collected as individual adverse events; in subsequent years, multiple symptoms of asthma exacerbation were
considered as a single adverse event. th Respiratory Adverse Events (All events reported at ≥3.0% in any year) a: Year 1 data only for subjects who enrolled for longer-term follow-up. Adverse events solicited from patient during multiple office visits in Year 1. In
subsequent years, adverse events solicited only at annual follow-up visit. b: Control group subjects exited from Study after Year 3 evaluations. c: Includes adverse events reported as “Respiratory Tract Infection” and “Lower Respiratory Tract Infection”. usage for asthma symptoms was comparable between the BT
and Control groups during Years 1, 2 and 3. number of subjects had been adjusted to reflect their
current level of asthma control. Compared to their base-
line pre-BT usage, over the course of the 5 years, an
average of 57% of BT subjects reported a decrease in
their LABA use, 40% of subjects reported no change in a: Control group subjects exited from Study after Year 3 evaluations.
b: p value from Fisher’s Exact test Maintenance Asthma Medication Use During the review of maintenance asthma medications
at each annual visit, the medication dosages for a Table 5 Summary of Healthcare Utilization Events
Hospitalizations
Percent of Subjects [95% CI]
(Number of Events)
Emergency Room Visits
Percent of Subjects [95% CI]
(Number of Events)
Total Number of Subjects
BT
Control
p-valueb
BT
Control
p-valueb
Year 1
BT = 45
Control = 24
6.7% [0.0, 14.0]
(3)
0
0.55
4.4% [0.0, 10.5]
(2)
0
0.54
Year 2
BT = 45
Control = 24
6.7% [0.0, 14.0]
(3)
0
0.55
6.7% [0.0, 14.0]
(3)
12.5% [0.0, 25.7]
(3)
0.41
Year 3
BT = 43
Control = 21
2.3% [0.0, 6.8]
(3)
4.8% [0.0, 13.9]
(1)
1.00
4.7% [0.0, 10.9]
(3)
4.8% [0.0, 13.9]
(3)
1.00
Year 4a
BT = 43
Control = 0
2.3% [0.0, 6.8]
(1)
–
9.3% [0.6, 18.0]
(6)
–
Year 5a
BT = 42
Control = 0
2.4% [0.0, 7.0]
(1)
–
4.8% [0.0, 11.2]
(2)
– Pulmonary Function Tests Pulmonary function tests were performed when subjects
were taking only ICS as their maintenance asthma medi-
cation (either after a 2 week withdrawal of LABAs for
subjects that were on ICS+LABA or for subjects on just
ICS). The mean post-bronchodilator FEV1 and FVC
values over time are presented graphically in Figure 1,
respectively. Both measures of lung function (FEV1 and
FVC) remained stable and showed no deterioration over
the 5 year period post-BT. Similarly, total lung capacity and residual volumes
remained stable out to 5 years in the BT group. There
were small improvements over baseline in Diffusion
Capacity in both the BT and Control groups over the
course of the study. a: Year 1 data only for subjects who enrolled for longer-term follow-up. b: Control group subjects exited from Study after Year 3 evaluations. c: Includes adverse events reported as “Asthma”, “Exacerbations of asthma”,
“Wheeze”, “Dyspnea”, and “Respiratory infection”. a: Year 1 data only for subjects who enrolled for longer-term follow-up. b: Control group subjects exited from Study after Year 3 evaluations. c: Includes adverse events reported as “Asthma”, “Exacerbations of asthma”,
“Wheeze”, “Dyspnea”, and “Respiratory infection”. There was an improvement over baseline in the metha-
choline PC20 doubling in the BT group compared to the
Control group in each year out to Year 3. The differences
between groups were statistically significant in Year 2
and Year 3, but not in Year 1 (methacholine PC20 dou-
blings BT versus Control: Year 1: 1.53 ± 2.29 vs 1.00 ±
2.46, p = 0.378; Year 2: 1.21 ± 2.99 vs -0.47 ± 2.31, p =
0.024; Year 3: 1.31 ± 2.96 vs -0.44 ± 2.27, p = 0.025). their LABA use, and 3% reported an increase in their
LABA use. In the Control group, over the course of the
3 years, an average of 54% of Control subjects reported
a decrease in their LABA use, 43% of subjects reported
no change in their LABA use, and 3% reported an
increase in their LABA use. In the same period an aver-
age of 49% of subjects in the BT group and 47% of sub-
jects in the Control group were no longer taking LABA
as controller medication. their LABA use, and 3% reported an increase in their
LABA use. Pulmonary Function Tests In the Control group, over the course of the
3 years, an average of 54% of Control subjects reported
a decrease in their LABA use, 43% of subjects reported
no change in their LABA use, and 3% reported an
increase in their LABA use. In the same period an aver-
age of 49% of subjects in the BT group and 47% of sub-
jects in the Control group were no longer taking LABA
as controller medication. Table 5 Summary of Healthcare Utilization Events Table 5 Summary of Healthcare Utilization Events Table 5 Summary of Healthcare Utilization Events Control group subjects exited from Study after Year 3 evaluations. Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Page 6 of 9 Table 6 Use of Oral Corticosteroid Pulses for Asthma
Symptoms
Oral Corticosteroid Pulses:
Events/Subject/Year
(Percent of Subjects)
Total Number of Subjects
BT
Control
Year 1a
BT = 45
Control = 24
0.60 (24.5%)c
0.42 (20.8%)c
Year 2
BT = 45
Control = 24
0.49 (24.5%)
0.54 (33.3%)
Year 3
BT = 43
Control = 21
0.33 (25.6%)
0.52 (23.8%)
Year 4b
BT = 43
Control = 0
0.63 (27.9%)
–
Year 5b
BT = 42
Control = 0
0.62 (30.9%)
–
a: Year 1 data only for subjects who enrolled for longer-term follow-up. b: Control group subjects exited from Study after Year 3 evaluations. c: Includes adverse events reported as “Asthma”, “Exacerbations of asthma”,
“Wheeze”, “Dyspnea”, and “Respiratory infection”. Table 6 Use of Oral Corticosteroid Pulses for Asthma
Symptoms that typically occur in patients with moderate and severe
asthma. Discussion The recently approved Alair System for bronchial ther-
moplasty provides a new treatment option for patients
with severe asthma that remain symptomatic despite
taking inhaled corticosteroids and long-acting-b2-
agonists, the current standard-of-care medications. Three randomized clinical trials in patients with differ-
ing severity of asthma have demonstrated the safety of
this device-based treatment out to one year [9-11]. In
the AIR Trial, at 12 months post-treatment, while
patients were on ICS alone, these benefits included sig-
nificantly greater improvements in the BT group than in
the Control group in mild exacerbation rates, morning
peak expiratory flow, scores on the AQLQ and ACQ,
the percentage of symptom-free days and symptom
scores, while fewer puffs of rescue medication were
required [9]. Each of these studies has also reported on
the short-term (treatment period) and long-term (out to
12 months) adverse event profile associated with the
bronchial thermoplasty. The reported increase in
respiratory adverse events in the short-term represented
the further aggravation of the worsening of asthma
symptoms that is associated with bronchoscopy in
patients with asthma [14]. During the long-term post-
treatment period out to 12 months, fewer subjects in
the BT group reported respiratory adverse events [9-11]. Data are now presented for the safety of this treatment
over a 5 year period. The mean reduction from baseline in ICS dose for BT
subjects was 182 μg/day (p = 0.09), 135 μg/day (p =
0.32), 150 μg/day (p = 0.25), 151 μg/day (p = 0.23), and
194 μg/day (p = 0.16) at Years 1, 2, 3, 4, and 5 respec-
tively (p-values from a Signed Rank test). At Year 3, the
mean reduction in the ICS daily dose in the Control
group was 112 μg/day. The reduction in ICS was not
significantly different between the BT group and the
Control group at years 2 and 3 (p = 0.93 for Year 2, and
p = 0.92 for Year 3), years when data were available for
both the BT and Control groups. At 3 years the propor-
tion of subjects in the BT group with changes from
baseline in their maintenance ICS dose was 27% with a
decrease, 56% with no change, and 17% with an
increase. The corresponding numbers for the Control
group were 29%, 52% and 19% respectively. Review of Serial (Annual) Chest X-rays Review of Serial (Annual) Chest X-rays 18 of the 45 BT subjects (40%), and 9 of the 24 Control
subjects (37%) had finding(s) noted on chest x-rays. Findings ranged from air trapping, pleural thickening,
increased density/consolidation, hyperinflation, nodules/
granuloma, increased vascular markings, and bronchial
wall thickening. None of the findings noted in either
group were clinically significant structural changes. The
findings noted were either pre-existing, minor and tran-
sient, or consistent with acute inter-current illnesses Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Page 7 of 9 Page 7 of 9 Figure 1 FEV1 and FVC over Time. Post-bronchodilator FEV1 and FVC over time. Data represent group mean values for each year. Note that
participation of the Control subjects in the study was terminated after Year 3 evaluation. Figure 1 FEV1 and FVC over Time. Post-bronchodilator FEV1 and FVC over time. Data represent group mean values for each year. Note that
participation of the Control subjects in the study was terminated after Year 3 evaluation. BT is a novel treatment that alters the amount of air-
way smooth muscle, and asthma is chronic disease that is
associated with structural changes in the airway wall, it
was important to know whether this treatment is asso-
ciated with any longer term adverse outcomes. The parti-
cipation of patients originally enrolled in the AIR Trial in
this additional 4 year follow-up study provides data to
support the longer-term safety of BT out to at least
5 years. The absence of serious events indicates that the
integrity of the airways is not compromised over the long
term as a result of BT treatment. This is supported by
the observation of no deterioration of baseline FEV1 or
FVC over this period, and the ability of the airways to
respond to bronchodilator administration. While the rate
of decline in lung function can vary in asthma, with some
subjects demonstrating equivalent declines to those with-
out airways disease, and others suffer an accelerated
decline [15-17], it is reassuring that following BT, there
was no significant loss of lung function over a 5 year per-
iod. The rate of occurrence of respiratory related adverse
events remained low and stable between Year 2 and Year
5 (1.1 to 1.3 events/subject/year) and comparable to the
Control group for Year 2 and Year 3 (1.2 and 1.3 events/
subject/year, respectively). Review of Serial (Annual) Chest X-rays The frequency and method of collecting adverse events in this study was different in
Year 1 compared to the other 4 years, resulting in a
higher rate of respiratory adverse events in both the BT
and Control groups in Year 1. During Year 1, the solicita-
tion of adverse events was more frequent (12 office visits
and 9 telephone contacts) compared to subsequent years
when adverse events were solicited once a year and could
be subject to recall bias. Also important is the fact that
during Year 1, multiple symptoms for a given adverse
event were recorded as individual adverse events as com-
pared to the subsequent years when multiple related
symptoms were recorded as a single event. Because of
these methodological issues, comparisons of adverse
event rates within each group for Year 1 and subsequent
years are not appropriate. The lower rates of respiratory
adverse events in Years 2 to 5 compared to Year 1 may
reflect some underreporting as a result of recall bias at
the yearly solicitation of adverse events. Every effort was
made to carefully solicit adverse events from study sub-
jects during annual evaluations and augmented with a
thorough review of medical records at participating insti-
tutions. Review of medical charts is lacking for instances
where a subject may have been treated for adverse event(s)
at a different institution. Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Page 8 of 9 Page 8 of 9 Nevertheless a high proportion of patients who received
BT (86%) participated in long-term follow-up, which
provides support for the generalizability of the safety
data. Thirdly, the solicitation of adverse events on a
yearly basis has the potential of under reporting due to
recall bias. However, the occurrence of major events
such as severe exacerbations (steroid pulses for asthma
symptoms), emergency room visits, and hospitalizations
is less frequent and less likely to be forgotten and there-
fore not reported. Medical charts were reviewed to ver-
ify steroid pulses prescribed for asthma symptoms
(severe exacerbations) and reported adverse events or to
identify adverse events that may not have been reported
by the subject, unless the subject had presented at a dif-
ferent medical facility for an adverse event during that
year. Review of Serial (Annual) Chest X-rays Finally, the study was not blinded and this may
add bias to the eliciting of adverse events. Despite these
limitations, over the 5 year follow-up of BT treated sub-
jects, the absence of unexpected respiratory adverse
events, no increases in hospitalizations or emergency
room visits for respiratory symptoms, and maintenance
of stable lung function demonstrated safety of BT out to
5 years. Measures such as ER Visits and hospitalizations for
respiratory symptoms are generally considered to be
important measures of safety, especially if an interven-
tion results in an increase in the rate of one or more of
these events. Consistent with the low rate of respiratory
adverse events is the stable incidence of healthcare utili-
zation events (hospitalizations and emergency room vis-
its for respiratory symptoms). There was no worsening
of the proportion of subjects in the BT group experien-
cing hospitalizations or ER visits for respiratory symp-
toms beyond Year 1 out to 5 years. The rates of ER
visits and the proportion of subjects with ER visits in
the Control group were comparable to the BT group
during Years 1, 2, and 3. Longer-term safety of BT is
supported further by our findings of no deterioration in
measurements of static and dynamic lung volumes or
diffusing capacity over the 5 years post-treatment. In
addition
airway
responsiveness
to
methacholine
remained stable in the BT group out to the last mea-
surement performed at year 3 post-treatment. Stable
lung function measures in this group of patients with
moderate to severe asthma support the previously
reported conclusion based on lung function measures
[12,13] and high resolution CT [13] that BT-treated air-
ways do not develop late scarring that could result in
narrowing of the airways [12]. Conclusions This report provides long-term safety data on BT in
patients in whom BT was performed by trained opera-
tors, and patients were carefully observed especially in
the first year. The absence of clinical complications
based on adverse event reporting, healthcare utilization
events, and the maintenance of stable lung function (no
deterioration of FEV1) over a 5-year period post-BT in
patients with moderate to severe asthma suggest long-
term safety of the procedure out to 5 years. Although x-rays are not sensitive enough for the pur-
pose of demonstrating structural abnormalities, a review
of the serial x-rays obtained annually did not reveal any
clinically significant findings. While the use of high
resolution computed tomography (HCRT) would have
been more informative, it was not performed in this
study. Bronchial hyperresponsiveness to methacholine
improved in the thermoplasty group in years 2 and 3,
but not in year 1. This is an interesting result in favour
of a long-term efficacy of the procedure. However, the
lack of follow-up of the control group on years 4 and 5
and the observational nature of the data limits the rele-
vance of this finding. Abbreviations
AE Ad
E AE: Adverse Event; ACQ: Asthma Control Questionnaire; AIR: Asthma
Intervention Research; AIR2: Asthma Intervention Research 2; AQLQ: Asthma
Quality of Life Questionnaire; BT: Bronchial thermoplasty; FEV1: Forced
Expiratory Volume in 1 second; FVC: Forced Vital Capacity; HRCT: High
resolution computed tomography; ICS: Inhaled corticosteroid; L: Liters; LABA:
Long-Acting-β2-Agonist; OCS: Oral corticosteroid; PC20: Provocative
Concentration causing 25% drop in FEV1; PEF: Peak Expiratory Flow; RISA:
Research in Severe Asthma; SD: Standard deviation There are several limitations to the study. Firstly, not
all patients enrolled in the Asthma Intervention
Research Trial participated in long-term follow-up,
although none of the reasons for non-participation was
due to mortality. Secondly, the follow-up period was
longer in patients who received BT (at 5 years) com-
pared to the Control group (3 years). Subjects who had
been randomized to the Control group in the predeces-
sor AIR Trial may have opted to pursue other treatment
options, and for those that agreed to the longer-term
follow up, it was deemed to be unethical [18] to require
them to withhold other alternative treatment options
(including new experimental treatments) for a period of
5 years in order to avoid confounding of data. Acknowledgements Members of the AIR Trial Study Group were as follows:
Co-Investigators and Study Coordinators – Brazil: Santa Casa: P.G. Cardoso, M. Cavalcanti, P.R.D. Soares, S. Zelmanovitz; Canada: McMaster
University: P. Nair, S. Goodwin, S. Keogh, M. Kjarsgard; Montreal Chest Institute:
J. Bourbeau, F. Houghton, R. Mangaser; London Health Science Centre: N. Patterson, S. Metha, J. Howard, L. MacBean; Laval University: S. Martel, L-P. Boulet, S. Savord, L. Morel, L. Trepanier; Denmark: Odense University Hospital:
F. Rasmussen, H.M. Christensen; United Kingdom: University of Glasgow: S. Bicknell, R. Chaudhuri, E. Hothersall, J. Lafferty, J. Jarvis; University of
Manchester: C. Prys-Picard, G. Fletcher, A. Fletcher; Newcastle University: B. Higgins, T. Small, B. Foggo, L. Mackay, S. Parker; University of Leicester: M. Berry, D Shaw, P. Haldar, A. Charalambou, M. Hopkin, A. Raj, N. Yousaf, N. Goodman. Page 9 of 9 Page 9 of 9 Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 Authors’ contributions IDP, M.D.: Contributed to the acquisition and interpretation of the data, and
gave final approval of the version to be published. DM, M.D.: Contributed to the acquisition and interpretation of the data, and
gave final approval of the version to be published. ML, M.D.: Contributed to the acquisition and interpretation of the data, and
gave final approval of the version to be published. NSS, PhD.: Contributed to the execution of the study, acquisition and
interpretation of the data, writing and revision of the manuscript, and gave
final approval of the version to be published. Had full access to the data
and will vouch for the integrity of data analysis, and the accuracy and
completeness of the reported data. 15. Sherrill D, Guerra S, Bobadilla A, Barbee R: The role of concomitant
respiratory diseases on the rate of decline in FEV1 among adult
asthmatics. Chest 2003, 21(1):95-10. 15. Sherrill D, Guerra S, Bobadilla A, Barbee R: The role of concomitant
respiratory diseases on the rate of decline in FEV1 among adult
asthmatics. Chest 2003, 21(1):95-10. 16. Lange P, Parner J, Vestbo J, Schnohr P, Jensen G: A 15-Year Follow-Up
Study of Ventilatory Function in Adults with Asthma. N Engl J Med 1998,
339(17):1194-200. 16. Lange P, Parner J, Vestbo J, Schnohr P, Jensen G: A 15-Year Follow-Up
Study of Ventilatory Function in Adults with Asthma. N Engl J Med 1998,
339(17):1194-200. 17. James AL, Palmer LJ, Kicic E, et al: Decline in Lung Function in the
Busselton Health Study: The Effects of Asthma and Cigarette Smoking. Am J Respir Crit Care Med 2005, 171(2):109-14. 17. James AL, Palmer LJ, Kicic E, et al: Decline in Lung Function in the
Busselton Health Study: The Effects of Asthma and Cigarette Smoking. Am J Respir Crit Care Med 2005, 171(2):109-14. 18. Rothman KJ, Michels KB: The continuing unethical use of placebo
controls. New Engl J Med 1994, 331:394-398. 18. Rothman KJ, Michels KB: The continuing unethical use of placebo
controls. New Engl J Med 1994, 331:394-398. Competing interests Neil C. Thomson, Adalberto S. Rubin, Robert M. Niven, Paul A. Corris, Hans
Christian Siersted, Ronald Olivenstein, Ian D. Pavord, David McCormack, Michel
Laviolette, Gerard Cox all received industry-sponsored grant funding from
Asthmatx, the manufacturers of the Alair® System, for participating in clinical
trials. Narinder S Shargill is an employee of Asthmatx. Received: 29 October 2010 Accepted: 11 February 2011
Published: 11 February 2011 Received: 29 October 2010 Accepted: 11 February 2011
Published: 11 February 2011 Author details
1 6. Serra-Battles J, Plaza V, Morejon E, Comella A, Brugues 1J: Costs of asthma
according to the degree of severity. Eur Respir J 1998, 12:1322-1326. 1Gartnavel General Hospital, University of Glasgow, Glasgow, UK. 2Irmandade
Santa Casa de Misericórdia da Porto Alegre, Brazil. 3University Hospital of
South Manchester and University of Manchester, Manchester, UK. 4Department of Respiratory Medicine, Freeman Hospital, Newcastle
University, Newcastle, UK. 5Odense University Hospital, Odense, Denmark. 6Montreal Chest Institute, McGill University, Montreal, Canada. 7Glenfield
General Hospital, University Hospitals of Leicester NHS Trust, Leicester, UK. 8London Health Sciences Center, Ontario, Canada. 9Laval Hospital, Laval
University, Quebec, Canada. 10Asthmatx, Inc., Sunnyvale, CA, US. 11St. Joseph’s Healthcare, McMaster University, Hamilton, Canada. 6. Serra-Battles J, Plaza V, Morejon E, Comella A, Brugues 1J: Costs of asthma
according to the degree of severity. Eur Respir J 1998, 12:1322-1326. 7. Antonicelli L, Bucca C, Neri M, et al: Asthma severity and medical resource
utilization. Eur Respir J 2004, 23:723-729. 8. Godard P, Chanez P, Siraudin L, Nicoloyannis N, Duru G: Costs of asthma
l t d
ith
it
E
R
i J 2002 19 61 67 7. Antonicelli L, Bucca C, Neri M, et al: Asthma severity and medical resource
utilization. Eur Respir J 2004, 23:723-729. 8. Godard P, Chanez P, Siraudin L, Nicoloyannis N, Duru G: Costs of asthma
are correlated with severity. Eur Respir J 2002, 19:61-67. 9. Cox G, Thomson NC, Sperb-Rubin A, the AIR Trial Study Group, et al:
Asthma Control During the Year after Bronchial Thermoplasty. New Engl
J Med 2007, 356:1327-1337. 9. Cox G, Thomson NC, Sperb-Rubin A, the AIR Trial Study G ,
, p
,
y
p,
Asthma Control During the Year after Bronchial Thermoplasty. New Engl
J Med 2007, 356:1327-1337. 10. Pavord ID, Cox G, Thomson NC, the RISA Trial Study Group, et al: Safety
and Efficacy of Bronchial Thermoplasty in Symptomatic, Severe Asthma. Am J Respir Crit Care Med 2007, 176:1185-1191. Pre-publication history y
The pre-publication history for this paper can be accessed here:
http://www.biomedcentral.com/1471-2466/11/8/prepub GC, M.B.: Contributed to the acquisition and interpretation of the data,
writing and revision of the manuscript, and gave final approval of the
version to be published. Had full access to the data and will vouch for the
integrity of data analysis, and the accuracy and completeness of the
reported data. The pre-publication history for this paper can be accessed here:
http://www.biomedcentral.com/1471-2466/11/8/prepub doi:10.1186/1471-2466-11-8
Cite this article as: Thomson et al.: Long-term (5 year) safety of
bronchial thermoplasty: Asthma Intervention Research (AIR) trial. BMC
Pulmonary Medicine 2011 11:8. Thomson et al. BMC Pulmonary Medicine 2011, 11:8
http://www.biomedcentral.com/1471-2466/11/8 The database for this study was managed by, and all statistical analyses
were performed by B. Armstrong, MS and J. Quiring, PhD (QST
Consultations, Ltd., Allendale, MI). Data and Safety Monitoring Board – W. Busse, MD, R. Schellenberg, MD,
Scott Berry, PhD, and A.S. Slutsky, MD (Chair)
The authors thank Michael Wechsler, MD (Brigham & Women’s Hospital,
Boston, MA) for his critical review of this manuscript prior to submission. Supported by: Asthmatx, Inc., Sunnyvale, CA The database for this study was managed by, and all statistical analyses
were performed by B. Armstrong, MS and J. Quiring, PhD (QST
Consultations, Ltd., Allendale, MI). The database for this study was managed by, and all statistical analyses
were performed by B. Armstrong, MS and J. Quiring, PhD (QST
Consultations, Ltd., Allendale, MI). Severe Asthma Research Program. J Allergy Clin Immunol 2007,
119:405-413. 3. Fuhlbrigge A, Adams R, Guilbert T, et al: The burden of asthma in the
United States. Am J Respir Crit Care Med 2002, 166:1044-1049. Data and Safety Monitoring Board – W. Busse, MD, R. Schellenberg, MD,
Scott Berry, PhD, and A.S. Slutsky, MD (Chair) Data and Safety Monitoring Board – W. Busse, MD, R. Schellenberg, MD
Scott Berry, PhD, and A.S. Slutsky, MD (Chair) 4. Schatz M, Zeiger R, Mosen D, Vollmer W: Asthma-specific quality of life
and subsequent asthma emergency hospital care. Am J Manag Care
2008, 14:206-211. The authors thank Michael Wechsler, MD (Brigham & Women’s Hospital,
Boston, MA) for his critical review of this manuscript prior to submission. Boston, MA) for his critical review of this manuscript prior to submission
Supported by: Asthmatx, Inc., Sunnyvale, CA Supported by: Asthmatx, Inc., Sunnyvale, CA 5. Vollmer W, Markson L, O’Connor E, Frazier E, Berger M, Buist AS:
Association of Asthma Control with Health Care Utilization - A
Prospective Evaluation. Am J Resp Crit Care Med 2002, 165:195-199. Received: 29 October 2010 Accepted: 11 February 2011
Published: 11 February 2011 2.
Moore W, Bleecker E, Curran-Everett D, et al: Characterization of the severe
asthma phenotype by the National Heart, Lung, and Blood Institute’s Authors’ contributions 11. Castro M, Rubin AS, Laviolette M, for the AIR2 Trial Study Group, et al:
Effectiveness and Safety of Bronchial Thermoplasty in the Treatment of
Severe Asthma: A Multicenter, Randomized, Double-blind, Sham-
controlled Clinical Trial. Am J Respir Crit Care Med 2010, 181:116-124. NCT, M.D.: Contributed to the acquisition and interpretation of the data,
writing and revision of the manuscript, and gave final approval of the
version to be published. Had full access to the data and will vouch for the
integrity of data analysis, and the accuracy and completeness of the
reported data. controlled Clinical Trial. Am J Respir Crit Care Med 2010, 181:116-12 12. Cox G, Miller J, Goodwin S, Fitzgerald JM, et al: Long-Term Follow-up of
Bronchial Thermoplasty for Asthma: Safety Results at 5 Years. Am J Respir
Crit Care Med 2008, 177:A567. ASR, M.D.: Contributed to the acquisition and interpretation of the data, and
gave final approval of the version to be published. ASR, M.D.: Contributed to the acquisition and interpretation of the data, and
gave final approval of the version to be published. ASR, M.D.: Contributed to the acquisition and interpretation of the data, and
gave final approval of the version to be published. RMN, M.D.: Contributed to the acquisition and interpretation of the data,
writing and revision of the manuscript, and gave final approval of the
version to be published. 13. Cox G, Laviolette M, Rubin A, Thomson N: Long Term Safety of Bronchial
Thermoplasty (BT): 3 Year Data from Multiple Studies. Am J Respir Crit
Care Med 2009, 179:A2780. RMN, M.D.: Contributed to the acquisition and interpretation of the data,
writing and revision of the manuscript, and gave final approval of the
version to be published. 14. Moore W, Murphy J, Calhoun W, Castro M, Chung F, Erzurum S, Jarjour N,
Wenzel S, Peters S, Bleecker E, NHLBI Severe Asthma Research Program
(SARP): Safety of investigative bronchoscopy in severe asthma. ATS 2006
Annual Meeting, San Diego, CA 2006. PAC, M.D.: Contributed to the acquisition and interpretation of the data, and
gave final approval of the version to be published. HCS, M.D.: Contributed to the acquisition and interpretation of the data, and
gave final approval of the version to be published. RO, M.D.: Contributed to the acquisition and interpretation of the data, and
gave final approval of the version to be published. References
l 1. National Health Interview Survey, National Center for Health Statistics. CDC 2008 [http://www.cdc.gov/nchs/fastats/asthma.htm], Accessed 2/10/10. CDC 2008 [http://www.cdc.gov/nchs/fastats/asthma.htm], Accessed 2/10/10. 2. Moore W, Bleecker E, Curran-Everett D, et al: Characterization of the severe
asthma phenotype by the National Heart, Lung, and Blood Institute’s
|
https://openalex.org/W2943126381
|
http://ejournal.unp.ac.id/index.php/sendratasik/article/download/103409/101306
|
Indonesian
| null |
TINJAUAN KOREOGRAFI TARI NGAYUN NUCI DI SEMURUP KECAMATAN AIR HANGAT KABUPATEN KERINCI PROVINSI JAMBI
|
Jurnal Sendratasik (edisi elektronik)
| 2,019
|
cc-by
| 3,908
|
Keywords: review of the choreography. Ngayun Nuci Dance Keywords: review of the choreography. Ngayun Nuci Dance D
Jurus
FBS Univer
Jurus
FBS Univer
e-mail: aiuu.le
Thi
ti l
i
t
d
ib D
Jurus
FBS Univer
Jurus
FBS Univer
e-mail: aiuu.le
Thi
ti l
i
t
d
ib Abstract This article aims to describe and explain about the reviews of Dance
Choreography Ngayun Nuci in Semurup Sub Regency of Kerinci Hagat Water Jambi
province. This type of research is a qualitative descriptive methods. The object of
research is the dance Nuci Ngayun. Instrument in this study is the researchers
themselves and assisted by mobile phone camera, voice recorder, and stationery. Types of data using primary data and secondary data. Engineering data collection
done by the study of librarianship, observation, interviews, and documentation. Technique of data analysis by collecting data, selecting data that is considered
important, compiling data that has already been selected, analyzed data with
interpretation techniques. The results of this research show that dance Ngayun
Nuci is a dance creation which is the development of a tradition of Nuci Ngayun
dance serves as the entertainment and aims to preserve traditional arts in
Semurup. Dance Dance-shaped Nuci Ngayun group that developed the science of
dance composition by koreografernya. Nuci Ngayun dance Creations rendered
with planning the choreography for pementasannya. And also this dance
movement is fixed at typical dance tradition. A. Pendahuluan Menurut Soedarsono (1986:81) bahwa tari adalah salah satu cabang kebudayaan
yang substansi materi bakunya adalah Gerak. Sedangkan Hawkins (1990:2) berpendapat
bahwa tari adalah ekspresi perasaan manusia yang diubah kedalam imajinasi dalam
bentuk media gerak sehingga sebagai ungkapan sipenciptanya. Menurt Supardjan (1980:54) tari kreasi baru di Indonesia pada umumnya masih
banyak yang bersumber dari materi tradisional. Tari Ngayun Nuci merupakan tari kreasi
yang masih bnayak bersumber dari tari tradisi karena dilihat dari unsur pokok suatu tari e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019 e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019 69 yaitu gerakannya yang masih monoton, dan unsur pendukung seperti kostum dan musik
yang digunakan masih sederhana Tari Ngayun Nuci tradisi adalah salah satu seni tari yang diwariskan oleh nenek
moyang yang pada awalnya bersifat ritual karena di dalam tarian itu terdapat unsur-
unsur magis. Adapun unsur-unsur megis dalam tarian Ngayun Nuci tak lain dan tak
bukan adalah untuk menyeru leluhur, bias juga diartikan sebagai ritual pengobatan,
termasuk bertujuan untuk tolak bala, melepas nazar dan rasa syukur atas panen padi
kepada pencipta. Mahyudin Dpt (selaku petinggi adat), mengungkapkan bahwa Kenduri Sko adalah
bentuk Upacara pengangkatan kepala adat dari suku saudara laki-laki ibu. Upacara ini
bertujuan untuk mengungkapkan rasa syukur atas berhasilnya panen raya. Saat panen
raya dalam Upacara Kenduri Sko ini, biasanya dihadikan berbagai kesenian masyarakat
tradisional setempat, mulai dari musik sampai kepada tarinya. Salah satu bentuk tarian
yang dipertunjukkan dalam Upacara Kenduri Sko ini adalah tari Ngayun Nuci, yang bisa
difungsikan sebagai tari ritual maupun tari seni pertunjukan dalam acara Kenduri Sko. Di
dalam Upacara adat Kenduri Sko ini tari Ngayun Nuci selalu ditampilkan karena tari
Ngayun Nuci ini berkaitan erat dengan ritual-ritual dan unsur-unsur Kenduri Sko. Tari
Ngayun Nuci dari dulu sapai zaman sekarang masih tetap ditarikan dan tidak pernah
ditinggalkan karena tari Ngayun Nuci ini merupakan salah satu bagian dari mantra-
mantra atau ritual-ritual dalam Upacara adat Kenduri Sko. A. Pendahuluan Dalam perkembangannya, nam-nama gerak pada tari Ngayun Nuci ada yang
berubah, penari yang dulu tidak beraturan jumlahnya dan sekarang terdiri dari 6 orang,
yang dulu tari Ngayun Nuci ditarikan pada saat Upacara Adat Kenduri Sko kemudian
sekarang ditarikan pada acara-acara hiburan, selanjutnya yang tradisional sebelum
melakukan tari ada syarat-syarat tertentu namun sekarang tidak ada lagi syarat-syarat
tersebut, pola lantai yang tidak beraturan sekarang sudah ditata dengan banyak bentuk
pola lantai, kostum pada penari wanita yang dahulu memakai baju adat berwarna merah
dengan bahan beludu memakia ikat pinggang berwarna kuning dari bahan songket,rok
dari songket berwarna kuning dan memakai kuluk sebagai penutup kepala namun pada
sekarang ini baju penari berwarna kuning yang dipadukan dengan rok yang berwarna
hitam gunting caina selutut yang ditempel manik-manik berwana kuning emas, memakai
ikat pinggang besi berwarna kuning emas lalu pada bawahannya memakai celana
berwarna coklat polos,pada bagian kepala wanita menggunakan ikat kepala yang hanya
untuk menutupi setengah kepala yang berwarna kuning diberi manik-manik yang
berjurai warna kuning dikepala diberi hiasan yang berbentuk kipas berwarna hijau putih
dan merah. Pada musik yang dahulu hanya pakai gendang sekarang sudah
dikembangkan dari gendang hingga seruling dan syair lagu. Namun pada saat sekarang
ini tari Ngayun Nuci ini sudah dikemas menjadi tari untuk acara pertunjukkan yaitu di
Gedung Nasional, di Vestifal Danau Kerinci dan di acara Pekan Budaya di Jambi. Nama-nama gerakan tari Ngayun Nuci yaitu : Gerakan tari Ngayun Nuci Tradisi
masih sangat sederhana hanya menggunakan 5 macam gerak yaitu,sembah meminta
ampun, angkat kaki mundam tikirap, lambai tangan mengelambai, ngayun silindang
dan sembah akhir.. Sedangkan pada tari Ngayun Nuci yang sudah di Kreasi sudah banyak
terjadi perubahan seperti pada gerak sudah menjadi 8 macam yaitu, sembah meminta
ampun, angkat kaki tunduk kapalo, tunduk kupalo, tunduk bandan munyembah,angkat
kaki guyang kupalo, mungajak munari, hentak kaki, dan nyampak punyakit. 70 e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019
70 e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019 70 Menurut Iskandar Zakaria selaku Koreografer tari Ngayun Nuci, tari Ngayun Nuci
ini di kemas atau di buat pada Tahun 2012 yang mana tari Ngayun Nuci di buat untuk
acara prtunjukkan, tetapi tetap bersumber dari tari Nagyun Nuci tradisional. Tari Ngayun
Nuci berfungsi untuk menghibur, maka dalam penampilannya idak menggunakan
persyaratan-persyaratan seperti tari Ngayun Nuci pada Upacara Kenduri Sko yang
bersifat ritual. Tari tersebut ditampilkan pada acara Festival Peduli Mayarakat Danau
Kerinci (FMPDK). B. Metode Penelitian Jenis penelitian ini adalah penelitian Kualitatif dengan metode deskriptif. Moleong
(2010:4)
menyatakanbahwa
penelitian
kualitatif
bersifat
deskriptif,
yang
akanmenyajikan data-data melalui kata-kata tertulis atau lisan dari orang-orang dan
perilaku yang diamati”. Instrumen dalam penelitian ini adalah peneliti sendiri sebagai
instrument kunci. Selain peneliti sebagai instrumen kunci, instrumen lain sebagai
pendukung yaitu alat pencatat, alat perekam dan alat audio visual lainnya seperti:
handphone, kamera digital. Jenis data dalam penelitian ini diklafikasikan dalam dua bentuk yaitu data primer
dan data sekunder. Data dalam penelitian dikumpulkan melalui studi pustaka, observasi,
wawancara, yang langsung dikumpul di daerah di mana ditemukannya tari Ngayun Nuci,
khususnya di daerah Air Hangat Kabupaten Kerinci Propinsi Jambi. Dan langkah-langkah
yang digunakan dalam menganalisa adalah pengumpulan data, reduksi data, penyajian
data dan penarikan kesimpulan. 1. Ide Garapan Tari Ngayun Nuci Kreasi
Ide Garapan tari Ngayun Nuci Krea Ide Garapan tari Ngayun Nuci Kreasi ini tercipta pada saat koreografer Iskandar
Zakaria di tuntut untuk membuat sebuah tari garapan baru dari Dinas Pariwisata untuk
acara pentas seni di Jambi dan Festival Peduli Masyarakt Danau Kerinci pada tahun
2012. Kemudian koreografer terinspirasi darigerak tari tradisi Ngayun Nuci yang ada di
Semurup. Kemudian Zakaria Iskandar menciptakan tari Ngayun Nuci berdasarkan
penglihatan Iskandar pada waktu acara Kenduri Sko di Semurup Iskandar merasa tertarik
untuk membuat ulang gerakan-gerakan tari tersebut. A. Pendahuluan Koreografi lebih diartikan sebagai pengetahuan penyusunan tari atau hasil
susunan tari, sedangkan seniman atau penyusunannya dikenal dengan nama
Koreografer, yang dalam bahasa sekarang lebih dikenal dengan penata tari (Sal
Murgiyanto 1983:3-4). Kemudian Soedarsono (1986:134) menyatakan bahwa
pengetahuan komposisi tari yang lazim disebut pengetahuan koreografi adalah
pengetahuan yang harus diketahui oleh seorang koreografer dari sejak menggarap
gerak-gerak tari sampai kepada pengetahuan tata cara menyiapkannya pada program
pertunjukan. 2. Proses Penggarapan Gerak tari Ngayun Nuci merupakan pengembangan gerak tari Ngayun Nuci
Tradisional, jumlah gerak tradisi yang dikembangkan 6 sedangkan gerak tradisi yang
masih tetap terdapat pada gerak tari kreasi 2 ragam gerak. Berikut ragam gerak yang
ada pada tari Kreasi: e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019 e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019 71 Tabel 1. Pengembangan Gerak Tari Ngayun Nuci Tradisi
ke Gerak Ngayun Nuci Kreasi
No
Gerak Tradisi
Gerak Kreasi
Keterangan
1. 2. 3. 4. 5. 6. 7. 8. Gerak duduk
butimpuh
Transisi 1
Gerak mungayun
badan
Gerak Mungharap
Gerak memohon
ampun
Gerak buhempeh
cembung
Transisi 2
Gerak mungajak
munari
Transisi 3
Gerak hentak kaki
Transisi 4
Gerak mungayun
busamo
Gerak Sembah Mintak
Ampun
Angkat Kaki Tunduk Kupalo
Gerak Tunduk kupalo
Gerak tunduk badan
munyembah
Gerak Angkat Kaki Guyang
Kupalo
Gerak Mungajak Munari
Gerak Hentak Kaki
Gerak Nyampak Punyakit. Dikembang
Dikembangkan
Dikembangkan
Dikembangkan
Dikembangkan
Tidak dikembangkan
Dikembangkan
Tidak
Dikembangkan
Berdasarkan tabel ditas bahwa bentu tari Ngayun Nuci Kreasi terdapat 6 ragam
gerak tradisional yang dikembangkan sedangkan gerak tari tradisi Ngayun Nuci terdapat Tabel 1. Pengembangan Gerak Tari Ngayun Nuci Tradisi
ke Gerak Ngayun Nuci Kreasi
No
Gerak Tradisi
Gerak Kreasi
Keterangan
1. 2. 3. 4. 5. 6. 7. 8. Gerak duduk
butimpuh
Transisi 1
Gerak mungayun
badan
Gerak Mungharap
Gerak memohon
ampun
Gerak buhempeh
cembung
Transisi 2
Gerak mungajak
munari
Transisi 3
Gerak hentak kaki
Transisi 4
Gerak mungayun
busamo
Gerak Sembah Mintak
Ampun
Angkat Kaki Tunduk Kupalo
Gerak Tunduk kupalo
Gerak tunduk badan
munyembah
Gerak Angkat Kaki Guyang
Kupalo
Gerak Mungajak Munari
Gerak Hentak Kaki
Gerak Nyampak Punyakit. Dikembang
Dikembangkan
Dikembangkan
Dikembangkan
Dikembangkan
Tidak dikembangkan
Dikembangkan
Tidak
Dikembangkan
Berdasarkan tabel ditas bahwa bentu tari Ngayun Nuci Kreasi terdapat 6 ragam
gerak tradisional yang dikembangkan sedangkan gerak tari tradisi Ngayun Nuci terdapat
2 ragam gerak tari Kreasi Ngayun Nuci. Namun tari Kreasi Ngayun Nuci juga terdapat
gerak transisi sebagai gerak penghubung dalam penampilan setiap gerak-gerak inti pada
gerak tari Kreasi Ngayun Nuci. Tabel 1. Pengembangan Gerak Tari Ngayun Nuci Tradisi
ke Gerak Ngayun Nuci Kreasi ngembangan Gerak Tari Ngayun Nuci Tradisi Berdasarkan tabel ditas bahwa bentu tari Ngayun Nuci Kreasi terdapat 6 ragam
gerak tradisional yang dikembangkan sedangkan gerak tari tradisi Ngayun Nuci terdapat
2 ragam gerak tari Kreasi Ngayun Nuci. 2. Proses Penggarapan Namun tari Kreasi Ngayun Nuci juga terdapat
gerak transisi sebagai gerak penghubung dalam penampilan setiap gerak-gerak inti pada
gerak tari Kreasi Ngayun Nuci. Berdasarkan tabel ditas bahwa bentu tari Ngayun Nuci Kreasi terdapat 6 ragam
gerak tradisional yang dikembangkan sedangkan gerak tari tradisi Ngayun Nuci terdapat
2 ragam gerak tari Kreasi Ngayun Nuci. Namun tari Kreasi Ngayun Nuci juga terdapat
gerak transisi sebagai gerak penghubung dalam penampilan setiap gerak-gerak inti pada
gerak tari Kreasi Ngayun Nuci. Berdasarkan tabel ditas bahwa bentu tari Ngayun Nuci Kreasi terdapat 6 ragam
gerak tradisional yang dikembangkan sedangkan gerak tari tradisi Ngayun Nuci terdapat
2 ragam gerak tari Kreasi Ngayun Nuci. Namun tari Kreasi Ngayun Nuci juga terdapat
gerak transisi sebagai gerak penghubung dalam penampilan setiap gerak-gerak inti pada
gerak tari Kreasi Ngayun Nuci. a. Duduk Butimpuh Dalam tari Ngayun Nuci tradisi, gerak Duduk Butimpuh bertujuan
untuk menunjukkan bahwa mereka akan memulai ritual menari setelah itu
merekapun mulai menari. Pada tari Ngayun Nuci Kreasi gerak ini berubah nama
menjadi gerak Sembah meminta ampun ini hanya sekedar sembah pembuka saja. b. Gerak Mungayun Badan dalam tari Ngayun Nuci tradisi gerak ini disebut Menari
sambil mengayunkan badan. Pada tari Ngayun Nuci Kreasi, gerak ini berubah nama
menjadi angkat kaki tunduk kepalo yang mana penarinya menggerakkan kaki
dengan tangan memegang luci dan mengayun kedua tangan ke kiri dan kanan. c. Gerak Mungharap dalam tari Ngayun Nuci tradisi gerak ini diartikan yang mana para
penari melakukan gerakan duduk dengan penuh harap. Sedangkan tari Ngayun Nuci
Kreasi berubah nama gerak menjadi tunduk badan munyembah dengan gerakan
duduk sambil memberi sembah. d. Gerak Buhempeh Cembung pada Ngayun Nuci tradisi yang mana gerakannya
meletakkan cembung di atas tanah dengan tangan bergerak keatas. Sedangkan pada
tari Ngayun Nuci Kreasi berubah nama gerak menjadi Angkat Kaki Guyang Kupalo
merupakan gerak dengan mengayun kedua tangan. e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019
72 e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019 e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019 72 e. Gerak Mungajak Munari Dalam tari Ngayun Nuci tradisi gerak Mungajak Munari ini
merupakan gerakkan penari melakukan gerakan yang gembira. Pada tari Ngayun
Nuci Kreasi nama tetap Mungajak munari. f. Gerak Hentak Kaki dalam tari Ngayun Nuci Tradisi merupakan gerakan yang mana
penari melakukan gerakan sambil menghentakan kakinya. Pada tari Ngayun Nuci
Kreasi namanya tidak berubah tetap gerak Hentak Kaki. g. 2) Desain Lantai Desain lantai merupakan garis yang dilalui oleh penari di atas lantai dan
formasi kelompok yang dibuat oleh penari dalam tari kelompok. Desain lantai dalam
Tari Ngayun Nuci terdiri dari garis lurus dan garis lingkaran dimana desain lantai
garis lurus berbanjar yang membentuk garis lurus berbanjar dan garis lingkaran
membentuk lingkaran penuh dan juga garis segitiga. 3. Koreografi tari Ngayun Nuci
a. Aspek Bentuk
1) Gerak
Gerak
tari
Ngayun Gerak tari Ngayun Nuci dominan dengan gerak maknawi yang
menyampaikan maksud masyarakat dalam meminta kesembuhan dan rasa syukur
kepada sang pencipta. Gerak dalam tari Ngayun Nuci merupakan pengembangan
dari gerak tari tradisi Ngayun Nuci yang ada Semurup dan gerak yang diciptakan
baru oleh koreografer tari Zakaria Iskandar. 2. Proses Penggarapan Gerak Ngayun Busamo dalam tari Ngayun nuci tradisi gerak ini ditarikan dengan cara
menaburkan beras yang mana bentuk dari rasa syukur atas melimpahnya hasil
panen padi. Sementara tari Ngayun Nuci Kreasi berubah nama menjadi Gerak
Nyampak Punyakit yang mana gerakannya serentak untuk mengakiri tari dengan
gerak ini. g. Gerak Ngayun Busamo dalam tari Ngayun nuci tradisi gerak ini ditarikan dengan cara
menaburkan beras yang mana bentuk dari rasa syukur atas melimpahnya hasil
panen padi. Sementara tari Ngayun Nuci Kreasi berubah nama menjadi Gerak
Nyampak Punyakit yang mana gerakannya serentak untuk mengakiri tari dengan
gerak ini. Tahapan yang dilakukan seorang Koreografi yaitu yang pertama Koreografer
mengumpulkan penari 6 cewek dan 1 cowok yang berfungsi sebagai pawang kemudian
Koreografer menerapkan tata cara latihan dan waktu yang dimiliki untuk latihan, waktu
latihan sebanyak 6 kali dalam satu minggu selama 1 bulan, kemudian Koreografer mulai
menjelaskan nama-nama gerakan yang akan di ajarkan nanti dan seterusnya Koreografer
mengajarkan atau mempraktekkan gerakan satu persatu kepada penarinya, lalu setelah
diajarkan Koreografer meihat kembali penari melakukan gerakan secara bersamaan dan
koreografernya melihat dari depan. Koreografer melihat dari hari ke hari penarinya
sudah menguasai apa yang dia praktekkan sehingga pada akhirnya waktu 1 bulan penuh
untuk latihan sudah berakhir dan Koreografer sudah siap untuk melakukan
pertunjukkan, namun sebelumnya di lakukan gladi resik terlebih dahulu sebelum
menampilkan hasil karya dari Koreografinya sehingga pada akhirnya tari Ngayun Nuci
yang sudah di Kreasikan sudah siap untuk ditampilkan. 3) Desain Dramatik Suasana dramatik pada tari Ngayun Nuci diawali dengan suasana tenang
dengan menceritakan tentang permohonan atau mintak ampun pada nenek moyang
dengan melakukan gerakan sembah mintak ampun dan gerakan angkat kaki tunduk
kupalo juga gerak tunduk kupalo. Selanjutnya pada bagian kedua suasana tari yaitu
tenang yang mana penari melakukan gerakan tunduk badan munyembah dengan
menceritakan permohonan ampun pada nenek moyang, kemudia dilanjutkan
dengan gerakan angkat kaki guyang kupalo dengan suasana tenang. Kemudian pada
bagian ke tiga dilanjutkan dengan gerakan mungajak munari dengan suasana 73
udian pada
n suasana e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019
73 e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019 e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019 e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019 73 gembira yang mana menceritakan tentang kegembiraan atas terkabulnya
permintaan. selanjutnya gerakan hentak kaki juga suasana gembira dan terakhir
dilakukan gerakan nyampak punyakit yang mana gerakan yang klimaks yaitu
mencertikan tentang kegembiraan atas kesembuhan yang dimintak pada nenek
moyang. 4) Komposisi Kelompok Komposisi kelompok dapat dibagi dua yaitu kelompok kecil dan kelompok
besar. Tari Ngayun Nuci termasuk komposisi kelompok besar karena terdiri dari 6
orang penari. 6) Kostum Kostum tari Ngayun Nuci terbuat dari kain tipis yang dasarnya mengkilap
yang warnanya kuning emas kemudian rok terbuat dari kain tipis juga yang dasarnya
mengkilap warnanya hitam dan kuning emas juga ikat pinggang ynag berwarna
merah terbuat dari dasar kain yang polos yang di modifikasi dengan kain polos
berwarna kuning emas. Kemudian ikat kepala terbuat dari kain dasar emas dengan
dilekatkan manik-manik pada bagian bawahnya. 7) Property 7) Property
La Meri (1986:106) Properti adalah benda-benda yang dipegang oleh
penari. Penggunaan properti tari harus mempertimbangkan jenis, fungsi, dan asas
pakai properti secara baik dan benar. Didalam Tari Ngayun Luci menggunakan
properti sapu lidi yang dihiasi dengan tirai. Dan nuci yang sudah di hias, kemudian
sabung yang berwarna merah,hijau dan putih. e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019
b. Aspek isi
1) ide
Menurut Zakaria Iskandar (wawancara, 10 Oktober 2018), terciptanya tari
Ngayun Nuci karena merupakan kebiasaan masyarakat Semurup dalam melakukan
Upacara Kenduri Sko yang dilakukan setiap 5 tahun sekali, yang menceritakan
tentang kegiatan adat masyarakat Semurup, yaitu kegiatan bertani dan cara
penyembuhan penyakit. 2) Suasana
Suasana pada tari Ngayun Nuci diawali dengan suasana tenang pada
gerakan Sembah mintak ampun dan menunjukan proses masyarakat dalam
mempersiapkan sesembahan dalam memintak izin memuji tempat. Kemudian
berangsur naik pada gerak angkat kaki tunduk kupalodan gerak tunduk kupalo, tapi
naik tidak terlalu tinggi dari suasana awal. Kemudian gerakan nya kembali tegang 5) Musik Musik dapat berfungsi untuk membentuk suasana. Dalam tari Ngayun Nuci,
musik menggambarkan suasana yang penuh dengan kekhusukan dan gembira. Suasana penuh kekhusukan tergambar pada saat penari dengan sungguh-sungguh
mengungkapkan permohonan pada roh nenek moyang dan diiringi dengan musik
yang tenang. Kemudian pada suasana gembira tergambar pada saat penari
mengungkapkan keberhasilannya dalam memohon dan diiringi dengan tempo musik
yang lebih cepat. Disamping musik, tari Ngayun Nuci juga diiringi dengan syair. Syair dan
diucapkan dalam bahasa daerah (kerinci). Musik dikemas dalam bentuk pertitur
musik. Alat musik yang digunakan dalam tari Ngayun Nuci adalah rebana, gendang
dan seruling. Aspek isi
1) ide Menurut Zakaria Iskandar (wawancara, 10 Oktober 2018), terciptanya tari
Ngayun Nuci karena merupakan kebiasaan masyarakat Semurup dalam melakukan
Upacara Kenduri Sko yang dilakukan setiap 5 tahun sekali, yang menceritakan
tentang kegiatan adat masyarakat Semurup, yaitu kegiatan bertani dan cara
penyembuhan penyakit. 2) Suasana
Suasana pada tari Ngayun Nuci diawali dengan suasana tenang pada
gerakan Sembah mintak ampun dan menunjukan proses masyarakat dalam
mempersiapkan sesembahan dalam memintak izin memuji tempat. Kemudian
berangsur naik pada gerak angkat kaki tunduk kupalodan gerak tunduk kupalo, tapi
naik tidak terlalu tinggi dari suasana awal. Kemudian gerakan nya kembali tegang 74
upalo, tapi
ali tegang e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019
74 e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019 e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019 dengan gerakan tunduk badan munyembahdengan menunjukan suasana penuh
dengan kekhusukan dalam memohon. Kemudian suasana meningkat lagi pada
gerakan angkat kaki guyang kupalo, mungajak munari, dengan kecepatan yang ikut
meningkat dan menunjukan suasana tegang karena permintaan masyarakat telah
dikabulkan. Kemudian suasana kembali tegang pada Gerakan hentak kakitetapi tidak
terlalu tegang dari gerak sebelumnya di mana pada gerakan ini menunjukkan bahwa
para penari terbawa suasana tari dengan musik yang cepat. Pada bagian akhir pada
gerak nyampak punyakit suasana kembali meningkat dengan kecepatan tempo
musik ikut meningkat. 4. Pembahasan Tari Ngayun Nuci merupakan tari Kreasi yang digarap oleh Koreografer dari
Semurup. Tari Ngayun Nuci digarap oleh Koreografer Zakaria Iskandar. Secara teori
koreografi tari yang diciptakan oleh Zakaria Iskandar telah memuat beberapa elemen
koreografi, seperti gerak, desain lantai, desain atas, komposisi kelompok, penari,iringan
tari dan kostum. Selain itu tari ini telah dirancang dengan pola tari kreasi yang
bersumberkan tari tradisional (Tari Ngayun Nuci). Tari Ngayun Nuci Kreasi juga
berangkat dari sumber tradisional Ngayun
Nuci, dan kemudian dikreasikan dalam
bentuk gerak baru. Maksudnya adalah tari kreasi Nagyun Nuci telah digarap melalui
proses garap koreografi, hal ini terlihat dari elemen-elemen koreografi yang terdapat
dalam tarian Ngayun Nnuci kreasi seperti telah dijelaskan sebelumnya. Setelah mengamati secara koreografi, tari Ngayun Nuci dapat dipastikan
merupakan sebuah tari kreasi baru yang terdiri dari aspek bentuk, dan isi. Tari Ngayun
Nuci tidak dapat dilepaskan dari adanya bentuk dan isi, kedua aspek ini memiliki peran
masing-masing dalam tari Ngayun Nuci. Apabila bentuk adalah ungkapan gagasan atau
ide tari secara visual yang dirasakan dan dipahami oleh penonton melalui ekspresi
penari dan suasana tarian tersebut. Kedua aspek ini saling mendukung satu sama lain. Oleh sebab itu, tari Ngayun Nuci memiliki isi yang disampaikan oleh penarinya melalui
ekspresi atau mimik wajahnya, sesuai dengan pendapat Sal Murgianto tersebut. Tari
Ngayun Nuci memiliki ide atau gagasan yang diungkapkan melalui gerak dan didukung
oleh ekspresi wajah penarinya serta suasana yang ditampilkan melalui pertunjukan teri
tersebut secara keseluruhan dari bagaian pertama sampai akhir pertunjukan. Berarti tari
Ngayun Nuci memiliki isi yang dapat menyampaikan ide yang diungkapkan melalui
suasana dan ekspresi dari tari Ngayun Nuci tersebut. D. Simpulan Lexy, Moleong, (1988). Metode Penelitian Kualitatif. Jakarta : Deperteman
Pendidikan dan Kebudayaan. Sal Murgianto
(1983)
Koreografi pengetahuan Dasar Komposisi Tari
Jakarta D. Simpulan Berdasarkan hasil penelitian yang telah peneliti uraikan, tari Ngayun Nuci Kreasi
merupakan tari yang dikembangkan dari tari tradisi Ngayun Nuci tradisi. Tari Ngayun
Nuci Kreasi menceritakan tentang para petani meminta supaya hasil panen padi semakin
banyak dan permohonan kesembuhan pada sang pencipta. Oleh sebab itu, dapat disimpulkan bahwa tari Ngayun Nuci kreasi tari kreasi yang
berebentuk tari kelompok yang menggunakan komposisi kelompok, serta digarap
dengan mengangkat tradisidari masyarakat Kabupaten Kerinci. Tari Nagyun Nuci Kreasi
merupakan pengembangan dari tari tradisi dengan pendekatan koreografi. Padatari
Ngayun Nuci Kreasi terdapat elemen-elemen komposisi tari yang terdiri dari tema,
gerak, desain atas,desain lantai, desain dramatik, desain musik, perlengkapan-
perlengkapan dan koreografi kelompok. Dimana tema mengangkat tradisi masyarakat 75
engkapan-
masyarakat e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019
75 e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019 e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019 75 Kerinci. Gerak dalam tari Nagyun Nuci Kreasi merupakan pengembangan dari gerak tari
tradisi. Desain lantai yang menunjukan cirikhas dari gerak tradisi yang memiliki kekuatan
dalam gerak yang kuat, pola lantai yang berbentuk segitiga menunjukkan pemujaan,
desain dramatik yang mengalir dari rendah ketinggi, musik yang digarap dengan syair-
syair pemuja kepada roh nenek moyang, proses penggarapan tari yang terstruktur oleh
penciptanya. perlengkapan-perlengkapan dalam tari pendukung suasana dalam tari dan
tema tari. Serta Koreografi kelompok yang menunjukkan nilai sosial masyarakat dalam
rasa syukur terhadap hasil panen padi. Oleh sebab itu, tari Ngayun Nuci Kreasi digarap dengan perencanaan koreografi
untuk pementasannya. Dan juga tari ini gerakannya tetap pada cirikhas tari tradisi. Berdsarkan kesimpulan maka disarankan: 1) perlu disarankan kepada lembaga
kebudayaan Pariwisata Kerinci Kabupaten Kerinci agar lebih memperhatikan tari tradisi
kerinci yang hampir punah agar bisa dikreasikan kembali dengan tidak meninggalkan ciri
khas dari tradisinya, untuk pertunjukan di dalam dan diluar Kabupaten Kerinci, 2)
diharapkan kepada generasi muda di Kabuapten Kerinci agar memeliharatari tradisi dan
tari Kreasi yang ada didalam Kabupaten Kerinci, 3) kepada seniman yang ada di
Kabupaten Kerinci agar lebih giat belajar pengetahuan Koreografi, agar karya-karya tari
yang diciptakan untuk mas mendatang lebih baik lagi, Sebab pengetahuan Koreografi
akan menuntun para Koreografer untuk menciptakan tari dengan cara yang sistematis. p
g
p
g
y
g
aftar Rujukan
Desfiarni. (2004). Tari Lukah Gilo Sebagai Rekaman Budaya Minangkabau Pra Islam:
dari Magis ke Seni Pertunjukan Hiburan.Yogyakarta: Kalika. Indrayuda . (2008). Tari Balance Madam pada masyarakat Nias Padang Sebuah
Perfektif Etnologi. Padang : UNP Press. Daftar Rujukan Desfiarni. (2004). Tari Lukah Gilo Sebagai Rekaman Budaya Minangkabau Pra Islam:
dari Magis ke Seni Pertunjukan Hiburan.Yogyakarta: Kalika. Indrayuda . (2008). Tari Balance Madam pada masyarakat Nias Padang Sebuah
Perfektif Etnologi. Padang : UNP Press. Lexy, Moleong, (1988). Metode Penelitian Kualitatif. Jakarta : Deperteman
Pendidikan dan Kebudayaan. Sal Murgianto. (1983). Koreografi pengetahuan Dasar Komposisi Tari. Jakarta
Depertemen Pendidikan dan Kebudayaan. Sal Murgiyanto. (1983). Koreografi Pengetahuan Dasar Komposisi Tari. Jakarta:
Depertemen Pendidikan dan Kebudayaan. Smith, Jacqueline. (1985). Komposisi tari Sebuah Pertunjukan Praktis Bagi Guru. Yogyakarta : Ikalasti. Soedarsono. (1977). Tari–tarian Indonesia. Jakarta: Proyek-proyek Pengembangan
Media Kebudayaan. e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019 e-jurnal Sendratasik
Vol. 7 No. 3 Seri B Maret 2019 76
|
https://openalex.org/W2793738750
|
https://dro.deakin.edu.au/articles/journal_contribution/The_ADAMTS5_metzincin_regulates_zebrafish_somite_differentiation/20809150/1/files/37066390.pdf
|
English
| null |
The ADAMTS5 Metzincin Regulates Zebrafish Somite Differentiation
|
International journal of molecular sciences
| 2,018
|
cc-by
| 11,158
|
AUTHOR(S) Carolyn Dancevic, Yann Gibert, Joachim Berger, Adam D Smith, Clifford Liongue, Nicole Stupka, Alister
Ward, Daniel R McCulloch 10536/DRO/DU:30110235 Downloaded from Deakin University’s Figshare repository
Deakin University CRICOS Provider Code: 00113B The ADAMTS5 metzincin regulates zebrafish somite differentiation Citation: Dancevic, Carolyn M, Gibert, Yann, Berger, Joachim, Smith, Adam D, Liongue, Clifford, Stupka,
Nicole, Ward, Alister C and McCulloch, Daniel R 2018, The ADAMTS5 metzincin regulates
zebrafish somite differentiation, International journal of molecular sciences, vol. 19, no. 3, Article
number: 766, pp. 1-12. Dancevic, Carolyn M, Gibert, Yann, Berger, Joachim, Smith, Adam D, Liongue, Clifford, Stupka,
Nicole, Ward, Alister C and McCulloch, Daniel R 2018, The ADAMTS5 metzincin regulates
zebrafish somite differentiation, International journal of molecular sciences, vol. 19, no. 3, Article
number: 766, pp. 1-12. DOI: http://www.dx.doi.org/10.3390/ijms19030766 DOI: http://www.dx.doi.org/10.3390/ijms19030766 © 2018, The Authors Reproduced by Deakin University under the terms of the Creative Commons Attribution Licence DOI: http://www.dx.doi.org/10.3390/ijms19030766 The ADAMTS5 Metzincin Regulates Zebrafish
Somite Differentiation Carolyn M. Dancevic 1,2, Yann Gibert 1,2, Joachim Berger 3 ID , Adam D. Smith 1,2,
Clifford Liongue 1,2 ID , Nicole Stupka 1,2 ID , Alister C. Ward 1,2,* and Daniel R. McCulloch 1,2 Carolyn M. Dancevic 1,2, Yann Gibert 1,2, Joachim Berger 3 ID , Adam D. Smith 1,2,
Clifford Liongue 1,2 ID , Nicole Stupka 1,2 ID , Alister C. Ward 1,2,* and Daniel R. McCulloch 1,2 1
School of Medicine, Deakin University, Waurn Ponds, Victoria 3216, Australia;
carolyn.dancevic@deakin.edu.au (C.M.D.); y.gibert@deakin.edu.au (Y.G.);
adam.smith@bio-strategy.com (A.D.S.); c.liongue@deakin.edu.au (C.L.);
nicole.stupka@deakin.edu.au (N.S.); daniel.mcculloch@uq.net.au (D.R.M.) 1
School of Medicine, Deakin University, Waurn Ponds, Victoria 3216, Australia;
carolyn.dancevic@deakin.edu.au (C.M.D.); y.gibert@deakin.edu.au (Y.G.);
adam.smith@bio-strategy.com (A.D.S.); c.liongue@deakin.edu.au (C.L.);
nicole.stupka@deakin.edu.au (N.S.); daniel.mcculloch@uq.net.au (D.R.M.) 3
Australian Regenerative Medicine Institute, Monash University, Clayton, Victoria 3800, Australia;
joachim.berger@monash.edu *
Correspondence: award@deakin.edu.au Received: 25 January 2018; Accepted: 1 March 2018; Published: 7 March 2018 Received: 25 January 2018; Accepted: 1 March 2018; Published: 7 March 2018 Abstract: The ADAMTS5 metzincin, a secreted zinc-dependent metalloproteinase, modulates the
extracellular matrix (ECM) during limb morphogenesis and other developmental processes. Here, the
role of ADAMTS5 was investigated by knockdown of zebrafish adamts5 during embryogenesis. This revealed impaired Sonic Hedgehog (Shh) signaling during somite patterning and early
myogenesis. Notably, synergistic regulation of myod expression by ADAMTS5 and Shh during
somite differentiation was observed. These roles were not dependent upon the catalytic activity of
ADAMTS5. These data identify a non-enzymatic function for ADAMTS5 in regulating an important
cell signaling pathway that impacts on muscle development, with implications for musculoskeletal
diseases in which ADAMTS5 and Shh have been associated. Keywords: metalloproteinase; extracellular matrix; ADAMTS; somite; muscle; zebrafish Downloaded from DRO:
http://hdl.handle.net/10536/DRO/DU:30110235 DRO
Deakin Research Online,
Deakin University’s Research Repository
Deakin University CRICOS Provider Code: 00113B Deakin University CRICOS Provider Code: 00113B International Journal of
Molecular Sciences International Journal of
Molecular Sciences International Journal of
Molecular Sciences Int. J. Mol. Sci. 2018, 19, 766; doi:10.3390/ijms19030766 1. Introduction The A Disintegrin-like and Metalloproteinase domain with Thrombospondin-1 motifs (ADAMTS)
metalloproteinases have important functions during developmental morphogenesis and are also
implicated in chronic disease. The proteoglycanase subfamily of ADAMTS1, 4, 5, 8, 9, 15 and
20 have broad functions, many attributed to their ability to remodel extracellular matrix (ECM)
components, such as the chondroitin sulphate proteoglycans versican and aggrecan. For example,
Adamts20 deficient bt/bt mice have defects in melanoblast survival [1] and Adamts9 haplo-insufficient
mice on an Adamts20 deficient (bt/bt) background present with a secondary cleft palate [2], in each
case associated with reduced versican proteolysis. Furthermore, ADAMTS1 has been implicated in
promoting atherosclerosis [3] and ADAMTS15 acts as a tumor suppressor in breast carcinoma [4],
potentially through proteoglycan proteolysis. However, non-enzymatic roles for several ADAMTS
family members have been described [5–7]. ADAMTS5 has been implicated in classic morphogenesis during development as well as in
chronic diseases such as arthritis and atherosclerosis. For example, combinatorial knockout of Adamts5,
Adamts9 and Adamts20 in mice prevented generation of bioactive fragments of versican that are
necessary for interdigital tissue apoptosis during development [8,9]. Adamts5 knockout mice also
developed myxomatous heart valves [10]. Furthermore, ADAMTS5 is considered one of the most
important aggrecan-degrading enzymes in arthritis [11,12] and may also promote lipoprotein binding
in atherosclerosis [13]. Int. J. Mol. Sci. 2018, 19, 766; doi:10.3390/ijms19030766 www.mdpi.com/journal/ijms Int. J. Mol. Sci. 2018, 19, 766 2 of 12 ECM remodeling is crucial to many developmental and disease processes, in part due to its role
in controlling cell signaling. Heparan sulphate proteoglycans bind fibroblast growth factors (FGFs),
thereby regulating their bioavailability to their receptors (FGFRs) [14] during developmental processes
such as myogenesis [15], as well as acting as co-receptors for Sonic Hedgehog (Shh) signaling [16]. A recent study identified Adamts9 as necessary for umbilical cord vascular development due, at least
in part, on its facilitation of Shh signaling [17]. Furthermore, levels of Hedgehog (Hh) signaling
correlate with the severity of osteoarthritis, which is potentially mediated by a pathway involving
ADAMTS5 [18]. Combined, these studies are suggestive of a complex interplay between the ECM and
crucial cell signaling pathways that involves ADAMTS proteoglycanases. This study identifies a role for ADAMTS5 during zebrafish embryogenesis. Abrogation of adamts5
expression disrupted Shh signaling during somite differentiation and reduced the expression of
the myogenic regulator myod. Importantly, somite differentiation was synergistically dependent
upon Shh and ADAMTS5. 2.1. The Secreted Metalloproteinase ADAMTS5 Is Expressed in Zebrafish Embryos We have previously elucidated a role for ADAMTS5 during myoblast fusion in post-natal
skeletal muscle from Adamts5−/−mice [19]. To investigate this further, zebrafish was employed
as a highly manipulable model of vertebrate development, which possesses a strongly conserved
adamts5 gene that is maternally inherited and then dynamically expressed in early-stage embryos [20]. To obtain a detailed understanding of ADAMTS5 protein expression in zebrafish, whole-mount
immunohistochemistry (IHC) was performed with a previously described anti-ADAMTS5 antibody
directed to its pro-domain [21], which is highly conserved in ADAMTS5 across vertebrates [20,22]. At 8 h post fertilization (hpf) (~80% epiboly), ADAMTS5 was strongly expressed in the dorsal
mesoendoderm at the animal pole with variable expression ventrally at the vegetal pole (Figure 1A). At 18 and 24 hpf, after the commencement of somitogenesis, ADAMTS5 was expressed in the rostral
neural tube (floor plate) and bilaterally in the prosencephalon (Figure 1A). 1. Introduction Moreover, these functions of ADAMTS5 were independent of catalytic
function. These data indicate that ADAMTS5 plays an important non-enzymatic role in regulating the
Shh pathway during embryogenesis that impacts on muscle development. This may be relevant in
conditions where ADAMTS proteins interact with the Shh signaling pathway, such as osteoarthritis and
umbilical cord vascular complications, as well as disorders where the myogenic program is disrupted,
such as muscular dystrophies. 2.2. Silencing of ADAMTS5 Expression To explore adamts5 function, the gene was targeted using two independent morpholino antisense
oligonucleotides (MOs) that were directed to either the AUG translation start site (AUG-MO) or the
splice site at the exon 2/3 boundary (2/3-MO) (Figure 1B), since exon 3 encodes for the catalytic
domain of ADAMTS5 in human, mouse and zebrafish [22]. ADAMTS5 protein expression was found
to be reduced upon adamts5 AUG-MO injection as shown by IHC and immunoblotting (Figure 1C). To confirm altered splicing of adamts5 transcripts after administration of the 2/3-MO, RT-PCR was
performed followed by sequencing analysis (Figure 1D). This indicated a 71% reduction of correctly
spliced adamts5 transcript and identified an alternate adamts5 transcript retaining the 569-bp intron
between exons 2 and 3 that results in inclusion of several premature stop codons (Figure 1D). The AUG-MO was subsequently used throughout the study to ensure translation of the entire gene
was disrupted, as well as to guarantee the maternal transcripts for this gene [20] were also affected;
however, similar data was obtained with the adamts5 2/3-MO [23]. 3 of 12 Int. J. Mol. Sci. 2018, 19, 766 Figure 1. Expression and silencing of adamts5 in zebrafish embryos. (A) ADAMTS5 expression in 8,
18 and 24 hpf wild-type embryos. Note strong early expression in the dorsal mesoendoderm (8 hpf,
arrows) and variable expression ventrally (8 hpf, arrowhead), with later expression in the floor plate
of the neural tube (18 and 24 hpf, arrows) and bilaterally in the prosencephalon (24 hpf, arrowheads). Asterisks = prosencephalon in no primary antibody control. Scale bar = 250 μm; (B) Schematic
representation of the adamts5 gene structure targeted with antisense morpholino oligonucleotides
(MO), and its subsequent splicing, indicating the primers used for RT-PCR and the size of the
resultant products; (C) Reduced ADAMTS5 expression is seen in adamts5 AUG-MO injected embryos
(asterisk) versus control (arrow) by whole-mount antibody labelling (left-hand panel) and Western
blot (right-hand panel) showing the 120 kDa ADAMTS5 species (asterisk) with a region of the
Coomassie blue stained gel shown below, demonstrating even loading; (D) RT-PCR of adamts5 mRNA
obtained from 24 hpf embryos following injection of the adamts5 2/3-MO at the 1-cell stage, showing
amplicons a and b (asterisk). β-actin was used as a house-keeping gene. Figure 1. Expression and silencing of adamts5 in zebrafish embryos. (A) ADAMTS5 expression in 8,
18 and 24 hpf wild-type embryos. 2.2. Silencing of ADAMTS5 Expression Note strong early expression in the dorsal mesoendoderm (8 hpf,
arrows) and variable expression ventrally (8 hpf, arrowhead), with later expression in the floor plate
of the neural tube (18 and 24 hpf, arrows) and bilaterally in the prosencephalon (24 hpf, arrowheads). Asterisks = prosencephalon in no primary antibody control. Scale bar = 250 µm; (B) Schematic
representation of the adamts5 gene structure targeted with antisense morpholino oligonucleotides
(MO), and its subsequent splicing, indicating the primers used for RT-PCR and the size of the
resultant products; (C) Reduced ADAMTS5 expression is seen in adamts5 AUG-MO injected embryos
(asterisk) versus control (arrow) by whole-mount antibody labelling (left-hand panel) and Western blot
(right-hand panel) showing the 120 kDa ADAMTS5 species (asterisk) with a region of the Coomassie
blue stained gel shown below, demonstrating even loading; (D) RT-PCR of adamts5 mRNA obtained
from 24 hpf embryos following injection of the adamts5 2/3-MO at the 1-cell stage, showing amplicons
a and b (asterisk). β-actin was used as a house-keeping gene. 2.2. Silencing of ADAMTS5 Expression Note strong early expression in the dorsal mesoendoderm (8 hpf,
arrows) and variable expression ventrally (8 hpf, arrowhead), with later expression in the floor plate
of the neural tube (18 and 24 hpf, arrows) and bilaterally in the prosencephalon (24 hpf, arrowheads). Asterisks = prosencephalon in no primary antibody control. Scale bar = 250 µm; (B) Schematic
representation of the adamts5 gene structure targeted with antisense morpholino oligonucleotides
(MO), and its subsequent splicing, indicating the primers used for RT-PCR and the size of the
resultant products; (C) Reduced ADAMTS5 expression is seen in adamts5 AUG-MO injected embryos
(asterisk) versus control (arrow) by whole-mount antibody labelling (left-hand panel) and Western blot
(right-hand panel) showing the 120 kDa ADAMTS5 species (asterisk) with a region of the Coomassie
blue stained gel shown below, demonstrating even loading; (D) RT-PCR of adamts5 mRNA obtained
from 24 hpf embryos following injection of the adamts5 2/3-MO at the 1-cell stage, showing amplicons
a and b (asterisk). β-actin was used as a house-keeping gene. Figure 1. Expression and silencing of adamts5 in zebrafish embryos. (A) ADAMTS5 expression in 8,
18 and 24 hpf wild-type embryos. Note strong early expression in the dorsal mesoendoderm (8 hpf,
arrows) and variable expression ventrally (8 hpf, arrowhead), with later expression in the floor plate
of the neural tube (18 and 24 hpf, arrows) and bilaterally in the prosencephalon (24 hpf, arrowheads). Asterisks = prosencephalon in no primary antibody control. Scale bar = 250 μm; (B) Schematic
representation of the adamts5 gene structure targeted with antisense morpholino oligonucleotides
(MO), and its subsequent splicing, indicating the primers used for RT-PCR and the size of the
resultant products; (C) Reduced ADAMTS5 expression is seen in adamts5 AUG-MO injected embryos
(asterisk) versus control (arrow) by whole-mount antibody labelling (left-hand panel) and Western
blot (right-hand panel) showing the 120 kDa ADAMTS5 species (asterisk) with a region of the
Coomassie blue stained gel shown below, demonstrating even loading; (D) RT-PCR of adamts5 mRNA
obtained from 24 hpf embryos following injection of the adamts5 2/3-MO at the 1-cell stage, showing
amplicons a and b (asterisk). β-actin was used as a house-keeping gene. Figure 1. Expression and silencing of adamts5 in zebrafish embryos. (A) ADAMTS5 expression in 8,
18 and 24 hpf wild-type embryos. 2.3. Notochord Morphology Is Perturbed in adamts5 Morphant Embryos
2.3. Notochord Morphology Is Perturbed in adamts5 Morphant Embryos Shh signaling from the notochord has been previously demonstrated to be important for adaxial
and paraxial mesoderm formation and myod expression during myogenesis [24], while no tail (ntl) is
an independent marker for axial mesoderm (notochord) [25]. Expression of shh and ntl remained
unchanged in 12 hpf adamts5 morphants compared to controls [23]. However, at 18 hpf the pattern of
shh (Figure 2A,D) and ntl (Figure 2B,E) staining was altered revealing disrupted notochord
morphology. Shh signaling from the notochord has been previously demonstrated to be important for adaxial
and paraxial mesoderm formation and myod expression during myogenesis [24], while no tail (ntl)
is an independent marker for axial mesoderm (notochord) [25]. Expression of shh and ntl remained
unchanged in 12 hpf adamts5 morphants compared to controls [23]. However, at 18 hpf the pattern of
shh (Figure 2A,D) and ntl (Figure 2B,E) staining was altered revealing disrupted notochord morphology. 2 4 Sk l t l M
l F
ti
I Di
t d i
d
t 5 M
h
t E b
2.4. Skeletal Muscle Formation Is Disrupted in adamts5 Morphant Embryos (A) Expression of shh in the notochord of 18 hpf control (a,a′, arrows) and adamts5 morphant
(b,b′, open arrowheads) embryos, with medio-lateral deviation in the adamts5 morphants (b,b′), with
anterior indicated (*); (B) Expression of ntl in the notochord of 18 hpf control (a,a′, arrows) and
adamts5 morphant (b,b′, open arrowheads) embryos, with medio-lateral deviation in the adamts5
morphants with anterior indicated (*); (C) Expression of myod in adaxial and paraxial mesoderm
of 18 hpf control embryos (a,a′) and its perturbation in adamts5 morphants (b,b′, open arrowheads)
with anterior indicated (*); (D) Quantitation of affected notochords in control and adamts5 morphant
embryos demarcated by shh in Figure 2A; (E) Quantitation of affected notochords in control and adamts5
morphant embryos demarcated by ntl in Figure 2B; (F) Quantitation of embryos with perturbed myod
expression in control and adamts5 morphant embryos demarcated in Figure 2C; (G) Double-transgenic
Tg(acta1:lifeact-GFP)/Tg(acta1:mCherryCaaX) embryos, in which thin filaments are marked green and
sarcolemma red, reveal loss of muscle integrity in 3 dpf adamts5 morphants. Muscle fibers of control
injected larvae feature the typical striation of the myofibril and regular myofibers within chevron-shape
somites, indicated by a dashed line (a). The boxed area in a is magnified in a′–a′′′. Myofibril striation
is partially lost within adamts5 morphants (arrowhead in b′) and the sarcolemma of the myofibers
disrupted (arrow in b′′). The boxed area in b is magnified in b′–b′′′. Scale bar = 50 µm. Figure 2. Notochord morphogenesis and muscle fiber formation is perturbed in adamts5 morphant
embryos. 2 4 Sk l t l M
l F
ti
I Di
t d i
d
t 5 M
h
t E b
2.4. Skeletal Muscle Formation Is Disrupted in adamts5 Morphant Embryos (A) Expression of shh in the notochord of 18 hpf control (a,a′, arrows) and adamts5 morphant
(b,b′, open arrowheads) embryos, with medio-lateral deviation in the adamts5 morphants (b,b′), with
anterior indicated (*); (B) Expression of ntl in the notochord of 18 hpf control (a,a′, arrows) and adamts5
morphant (b,b′, open arrowheads) embryos, with medio-lateral deviation in the adamts5 morphants
with anterior indicated (*); (C) Expression of myod in adaxial and paraxial mesoderm of 18 hpf control
embryos (a,a′) and its perturbation in adamts5 morphants (b,b′, open arrowheads) with anterior
indicated (*); (D) Quantitation of affected notochords in control and adamts5 morphant embryos
demarcated by shh in Figure 2A; (E) Quantitation of affected notochords in control and adamts5
morphant embryos demarcated by ntl in Figure 2B; (F) Quantitation of embryos with perturbed myod
expression in control and adamts5 morphant embryos demarcated in Figure 2C; (G) Double-
transgenic Tg(acta1:lifeact-GFP)/Tg(acta1:mCherryCaaX) embryos, in which thin filaments are marked
green and sarcolemma red, reveal loss of muscle integrity in 3 dpf adamts5 morphants. Muscle fibers
of control injected larvae feature the typical striation of the myofibril and regular myofibers within
chevron-shape somites, indicated by a dashed line (a). The boxed area in a is magnified in a′–a′′′. Myofibril striation is partially lost within adamts5 morphants (arrowhead in b′) and the sarcolemma
of the myofibers disrupted (arrow in b′′) The boxed area in b is magnified in b′–b′′′ Scale bar = 50
Figure 2. Notochord morphogenesis and muscle fiber formation is perturbed in adamts5 morphant
embryos. 2 4 Sk l t l M
l F
ti
I Di
t d i
d
t 5 M
h
t E b
2.4. Skeletal Muscle Formation Is Disrupted in adamts5 Morphant Embryos 2018, 19, 766 partially lost and the sarcolemma appeared irregular, indicating disrupted muscle organization
(Figure 2Gb–b′′′). adamts5 morphants was partially lost and the sarcolemma appeared irregular, indicating disrupted
muscle organization (Figure 2Gb–b′′′). rtially lost and the sarcolemma appeared irregular, indicating disrupted muscle organizat
gure 2Gb–b′′′). amts5 morphants was partially lost and the sarcolemma appeared irregular, indicating disrup
uscle organization (Figure 2Gb–b′′′). Figure 2. Notochord morphogenesis and muscle fiber formation is perturbed in adamts5 morphant
embryos. (A) Expression of shh in the notochord of 18 hpf control (a,a′, arrows) and adamts5 morphant
(b,b′, open arrowheads) embryos, with medio-lateral deviation in the adamts5 morphants (b,b′), with
anterior indicated (*); (B) Expression of ntl in the notochord of 18 hpf control (a,a′, arrows) and adamts5
morphant (b,b′, open arrowheads) embryos, with medio-lateral deviation in the adamts5 morphants
with anterior indicated (*); (C) Expression of myod in adaxial and paraxial mesoderm of 18 hpf control
embryos (a,a′) and its perturbation in adamts5 morphants (b,b′, open arrowheads) with anterior
indicated (*); (D) Quantitation of affected notochords in control and adamts5 morphant embryos
demarcated by shh in Figure 2A; (E) Quantitation of affected notochords in control and adamts5
morphant embryos demarcated by ntl in Figure 2B; (F) Quantitation of embryos with perturbed myod
expression in control and adamts5 morphant embryos demarcated in Figure 2C; (G) Double-
transgenic Tg(acta1:lifeact-GFP)/Tg(acta1:mCherryCaaX) embryos, in which thin filaments are marked
green and sarcolemma red, reveal loss of muscle integrity in 3 dpf adamts5 morphants. Muscle fibers
of control injected larvae feature the typical striation of the myofibril and regular myofibers within
chevron-shape somites, indicated by a dashed line (a). The boxed area in a is magnified in a′–a′′′. Myofibril striation is partially lost within adamts5 morphants (arrowhead in b′) and the sarcolemma
of the myofibers disrupted (arrow in b′′). The boxed area in b is magnified in b′–b′′′. Scale bar = 50
Figure 2. Notochord morphogenesis and muscle fiber formation is perturbed in adamts5 morphant
embryos. 2 4 Sk l t l M
l F
ti
I Di
t d i
d
t 5 M
h
t E b
2.4. Skeletal Muscle Formation Is Disrupted in adamts5 Morphant Embryos 2 4 Skeletal Mu le Fo
atio
I Di u ted i
ada t 5 Mo
ha t E b yo
2.4. Skeletal Muscle Formation Is Disrupted in adamts5 Morphant Embryos 2.4. Skeletal Muscle Formation Is Disrupted in adamts5 Morphant Embryos
Notochord perturbation is linked with defective somitic muscle formation and morphogenesis
[24]. Therefore, the disrupted notochord morphology in the adamts5 morphants suggested that
skeletal muscle development might be affected. This is also consistent with previous observations
indicating a skeletal muscle developmental defect in Adamts5 knockout mice [19]. Reduced or absent
paraxial mesodermal myod expression was also observed at 18 hpf (Figure 2C,F). To analyze potential
myofiber defects, adamts5 AUG-MO was administered to double-transgenic embryos, in which
myofiber thin filaments were labeled with Lifeact-GFP whereas the sarcolemma and t-tubules of the
myofiber were marked with mCherryCaaX via the CaaX-tag [26]. In control injected 3 dpf double-
transgenic larvae, Lifeact-GFP revealed the typical striation of the highly organized myofibril and
mCherryCaaX indicated regularly spaced t-tubules and ordered fiber membranes within chevron-
shaped somites (Figure 2Ga–a′′′). In contrast, the somites of adamts5 morphants were U-shaped,
which resembled a phenotype previously reported in shh mutant embryos [27] (Figure 2G(b)),
fi
i
hh
il bilit
t
ti l
I
dditi
fib il t i ti
ithi
fib
f
Notochord perturbation is linked with defective somitic muscle formation and morphogenesis [24]. Therefore, the disrupted notochord morphology in the adamts5 morphants suggested that skeletal
muscle development might be affected. This is also consistent with previous observations indicating
a skeletal muscle developmental defect in Adamts5 knockout mice [19]. Reduced or absent paraxial
mesodermal myod expression was also observed at 18 hpf (Figure 2C,F). To analyze potential myofiber
defects, adamts5 AUG-MO was administered to double-transgenic embryos, in which myofiber thin
filaments were labeled with Lifeact-GFP whereas the sarcolemma and t-tubules of the myofiber
were marked with mCherryCaaX via the CaaX-tag [26]. In control injected 3 dpf double-transgenic
larvae, Lifeact-GFP revealed the typical striation of the highly organized myofibril and mCherryCaaX
indicated regularly spaced t-tubules and ordered fiber membranes within chevron-shaped somites
(Figure 2Ga–a′′′). In contrast, the somites of adamts5 morphants were U-shaped, which resembled a
phenotype previously reported in shh mutant embryos [27] (Figure 2G(b)), confirming shh availability
as a potential cause. In addition, myofibril striation within myofibers of adamts5 morphants was 4 of 12
f Int. J. Mol. Sci. 2 4 Sk l t l M
l F
ti
I Di
t d i
d
t 5 M
h
t E b
2.4. Skeletal Muscle Formation Is Disrupted in adamts5 Morphant Embryos (A) Expression of shh in the notochord of 18 hpf control (a,a′, arrows) and adamts5 morphant
(b,b′, open arrowheads) embryos, with medio-lateral deviation in the adamts5 morphants (b,b′), with
anterior indicated (*); (B) Expression of ntl in the notochord of 18 hpf control (a,a′, arrows) and
adamts5 morphant (b,b′, open arrowheads) embryos, with medio-lateral deviation in the adamts5
morphants with anterior indicated (*); (C) Expression of myod in adaxial and paraxial mesoderm
of 18 hpf control embryos (a,a′) and its perturbation in adamts5 morphants (b,b′, open arrowheads)
with anterior indicated (*); (D) Quantitation of affected notochords in control and adamts5 morphant
embryos demarcated by shh in Figure 2A; (E) Quantitation of affected notochords in control and adamts5
morphant embryos demarcated by ntl in Figure 2B; (F) Quantitation of embryos with perturbed myod
expression in control and adamts5 morphant embryos demarcated in Figure 2C; (G) Double-transgenic
Tg(acta1:lifeact-GFP)/Tg(acta1:mCherryCaaX) embryos, in which thin filaments are marked green and
sarcolemma red, reveal loss of muscle integrity in 3 dpf adamts5 morphants. Muscle fibers of control
injected larvae feature the typical striation of the myofibril and regular myofibers within chevron-shape
somites, indicated by a dashed line (a). The boxed area in a is magnified in a′–a′′′. Myofibril striation
is partially lost within adamts5 morphants (arrowhead in b′) and the sarcolemma of the myofibers
disrupted (arrow in b′′). The boxed area in b is magnified in b′–b′′′. Scale bar = 50 µm. Int. J. Mol. Sci. 2018, 19, 766 5 of 12 To further analyze myofiber differentiation, myod expression was examined. Reduced or absent
paraxial mesodermal myod expression was observed at 12 hpf (Figure 3A(a,b)), whereas expression of
adaxial mesodermal myod was largely unaffected (Figure 3A(a,b)). Similar observations were made
with the adamts5 2/3-MO (Supplementary Figure S1) or upon co-injection of a p53 morpholino with
the adamts5 AUG-MO (Supplementary Figure S2B). To ensure that the specificity of the phenotype was
due to reduced adamts5 expression, mRNA encoding either wild-type or catalytically-inactive (E411A)
ADAMTS5 were co-injected, with both able to partially rescue the reduced paraxial mesodermal myod
expression (Figure 3A(c,d), respectively, and Figure 3B). This indicated that the enzymatic function of
ADAMTS5 was not necessary to induce the reduced myod expression. Int. J. Mol. Sci. 2018, 19, x FOR PEER REVIEW
5 of 12
of adaxial mesodermal myod was largely unaffected (Figure 3A(a,b)). 2 4 Sk l t l M
l F
ti
I Di
t d i
d
t 5 M
h
t E b
2.4. Skeletal Muscle Formation Is Disrupted in adamts5 Morphant Embryos Similar observations were made
with the adamts5 2/3-MO (Supplementary Figure S1) or upon co-injection of a p53 morpholino with
the adamts5 AUG-MO (Supplementary Figure S2B). To ensure that the specificity of the phenotype
was due to reduced adamts5 expression, mRNA encoding either wild-type or catalytically-inactive
(E411A) ADAMTS5 were co-injected, with both able to partially rescue the reduced paraxial
mesodermal myod expression (Figure 3A(c,d), respectively, and Figure 3B). This indicated that the
enzymatic function of ADAMTS5 was not necessary to induce the reduced myod expression. Figure 3. Loss of paraxial mesodermal myod expression in adamts5 morphant embryos. (A) Expression
of adaxial and paraxial myod in 12 hpf embryos injected with control MO (a, arrowheads), with
adamts5 morphant embryos showing substantial loss of paraxial expression (b, open arrowheads), as
well as mild loss of paraxial myod expression (b, open arrowheads). Rescue of paraxial myod
expression in adamts5 morphants co-injected with mRNA encoding wild-type (c, arrows) or
catalytically-inactive E411A (d, arrows) ADAMTS5. Control embryos injected with ADAMTS5 mRNA
encoding wild-type ADAMTS5 show unaffected myod expression in paraxial mesoderm (e, arrows). Scale bar = 100 μm; (B) Quantitation of embryos showing present or absent myod patterning
represented in (A). Figure 3. Loss of paraxial mesodermal myod expression in adamts5 morphant embryos. (A) Expression
of adaxial and paraxial myod in 12 hpf embryos injected with control MO (a, arrowheads), with adamts5
morphant embryos showing substantial loss of paraxial expression (b, open arrowheads), as well as
mild loss of paraxial myod expression (b, open arrowheads). Rescue of paraxial myod expression in
adamts5 morphants co-injected with mRNA encoding wild-type (c, arrows) or catalytically-inactive
E411A (d, arrows) ADAMTS5. Control embryos injected with ADAMTS5 mRNA encoding wild-type
ADAMTS5 show unaffected myod expression in paraxial mesoderm (e, arrows). Scale bar = 100 µm;
(B) Quantitation of embryos showing present or absent myod patterning represented in (A). Figure 3. Loss of paraxial mesodermal myod expression in adamts5 morphant embryos. (A) Expression
of adaxial and paraxial myod in 12 hpf embryos injected with control MO (a, arrowheads), with
adamts5 morphant embryos showing substantial loss of paraxial expression (b, open arrowheads), as
well as mild loss of paraxial myod expression (b, open arrowheads). Rescue of paraxial myod
expression in adamts5 morphants co-injected with mRNA encoding wild-type (c, arrows) or
catalytically-inactive E411A (d, arrows) ADAMTS5. Control embryos injected with ADAMTS5 mRNA
encoding wild-type ADAMTS5 show unaffected myod expression in paraxial mesoderm (e, arrows). 2 4 Sk l t l M
l F
ti
I Di
t d i
d
t 5 M
h
t E b
2.4. Skeletal Muscle Formation Is Disrupted in adamts5 Morphant Embryos Scale bar = 100 μm; (B) Quantitation of embryos showing present or absent myod patterning
represented in (A). Figure 3. Loss of paraxial mesodermal myod expression in adamts5 morphant embryos. (A) Expression
of adaxial and paraxial myod in 12 hpf embryos injected with control MO (a, arrowheads), with adamts5
morphant embryos showing substantial loss of paraxial expression (b, open arrowheads), as well as
mild loss of paraxial myod expression (b, open arrowheads). Rescue of paraxial myod expression in
adamts5 morphants co-injected with mRNA encoding wild-type (c, arrows) or catalytically-inactive
E411A (d, arrows) ADAMTS5. Control embryos injected with ADAMTS5 mRNA encoding wild-type
ADAMTS5 show unaffected myod expression in paraxial mesoderm (e, arrows). Scale bar = 100 µm;
(B) Quantitation of embryos showing present or absent myod patterning represented in (A). 2.5. Receptor-Mediated Sonic Hedgehog Signaling Is Affected in adamts5 Morphants
W
h
h
d h
d
d ADAMTS
ld l
d
l
d
l
2.5. Receptor-Mediated Sonic Hedgehog Signaling Is Affected in adamts5 Morphants 2.5. Receptor-Mediated Sonic Hedgehog Signaling Is Affected in adamts5 Morphants
h
h
d h
d
d A A
ld l
d
l
d
l
2.5. Receptor-Mediated Sonic Hedgehog Signaling Is Affected in adamts5 Morphants These experiments collectively suggest an interaction between
ADAMTS5 and Shh, such that they act synergistically to stimulate myod expression in adaxial
mesoderm (Figure 6). Int. J. Mol. Sci. 2018, 19, x FOR PEER REVIEW
6 of 12
adaxial expression of myod (Figure 4A(j–l),B) compared to untreated adamts5 morphant embryos
(Figure 4A(d–f),B). In a reciprocal experiment, the Smo agonist, SAG, was used to confirm the
dependency of Shh signaling on adamts5 expression. Administration of SAG on wild-type embryos
disrupted paraxial myod expression in a similar manner to adamts5 morphants (Figure 5A(d–I),B). However, the same concentration of SAG partially rescued the loss of paraxial myod patterning in the
adamts5 morphants (Figure 5A(g–l),B). These experiments collectively suggest an interaction between Figure 4. Combinatorial inhibition of Shh signaling and adamts5 disrupt paraxial and adaxial myod
expression. (A) Adaxial and paraxial myod expression in 12 hpf embryos treated with vehicle control
(a–c, arrows denote paraxial myod expression), vehicle + adamts5-MO (d–f, arrows denote absent
paraxial myod expression), 5 μM cyclopamine (g–i, arrowheads represent similar paraxial myod
staining compared to control group) and 5 μM cyclopamine + adamts5-MO (j–l, open arrowheads
represent absent adaxial myod expression and arrowhead represents absent paraxial myod staining
compared to adamts5 MO group). Scale bar = 200 μm; (B) Quantitation of embryos showing present
or absent myod patterning represented in (A). Figure 4. Combinatorial inhibition of Shh signaling and adamts5 disrupt paraxial and adaxial myod
expression. (A) Adaxial and paraxial myod expression in 12 hpf embryos treated with vehicle control
(a–c, arrows denote paraxial myod expression), vehicle + adamts5-MO (d–f, arrows denote absent
paraxial myod expression), 5 µM cyclopamine (g–i, arrowheads represent similar paraxial myod staining
compared to control group) and 5 µM cyclopamine + adamts5-MO (j–l, open arrowheads represent
absent adaxial myod expression and arrowhead represents absent paraxial myod staining compared to
adamts5 MO group). Scale bar = 200 µm; (B) Quantitation of embryos showing present or absent myod
patterning represented in (A). ADAMTS5 and Shh, such that they act synergistically to stimulate myod expression in adaxial
mesoderm (Figure 6). Figure 4. Combinatorial inhibition of Shh signaling and adamts5 disrupt paraxial and adaxial myod
expression. 2.5. Receptor-Mediated Sonic Hedgehog Signaling Is Affected in adamts5 Morphants
h
h
d h
d
d A A
ld l
d
l
d
l
2.5. Receptor-Mediated Sonic Hedgehog Signaling Is Affected in adamts5 Morphants (A) Adaxial and paraxial myod expression in 12 hpf embryos treated with vehicle control
(a–c, arrows denote paraxial myod expression), vehicle + adamts5-MO (d–f, arrows denote absent
paraxial myod expression), 5 μM cyclopamine (g–i, arrowheads represent similar paraxial myod
staining compared to control group) and 5 μM cyclopamine + adamts5-MO (j–l, open arrowheads
represent absent adaxial myod expression and arrowhead represents absent paraxial myod staining
compared to adamts5 MO group). Scale bar = 200 μm; (B) Quantitation of embryos showing present
or absent myod patterning represented in (A) y
y
g
y
ure 6). re 6) Figure 4. Combinatorial inhibition of Shh signaling and adamts5 disrupt paraxial and adaxial myod
expression. (A) Adaxial and paraxial myod expression in 12 hpf embryos treated with vehicle control
(a–c, arrows denote paraxial myod expression), vehicle + adamts5-MO (d–f, arrows denote absent
paraxial myod expression), 5 μM cyclopamine (g–i, arrowheads represent similar paraxial myod
staining compared to control group) and 5 μM cyclopamine + adamts5-MO (j–l, open arrowheads
represent absent adaxial myod expression and arrowhead represents absent paraxial myod staining
compared to adamts5 MO group). Scale bar = 200 μm; (B) Quantitation of embryos showing present
or absent myod patterning represented in (A). Figure 4. Combinatorial inhibition of Shh signaling and adamts5 disrupt paraxial and adaxial myod
expression. (A) Adaxial and paraxial myod expression in 12 hpf embryos treated with vehicle control
(a–c, arrows denote paraxial myod expression), vehicle + adamts5-MO (d–f, arrows denote absent
paraxial myod expression), 5 µM cyclopamine (g–i, arrowheads represent similar paraxial myod staining
compared to control group) and 5 µM cyclopamine + adamts5-MO (j–l, open arrowheads represent
absent adaxial myod expression and arrowhead represents absent paraxial myod staining compared to
adamts5 MO group). Scale bar = 200 µm; (B) Quantitation of embryos showing present or absent myod
patterning represented in (A). Figure 4. Combinatorial inhibition of Shh signaling and adamts5 disrupt paraxial and adaxial myod
expression. (A) Adaxial and paraxial myod expression in 12 hpf embryos treated with vehicle control
(a–c, arrows denote paraxial myod expression), vehicle + adamts5-MO (d–f, arrows denote absent
paraxial myod expression), 5 μM cyclopamine (g–i, arrowheads represent similar paraxial myod
staining compared to control group) and 5 μM cyclopamine + adamts5-MO (j–l, open arrowheads
represent absent adaxial myod expression and arrowhead represents absent paraxial myod staining
compared to adamts5 MO group). 2.5. Receptor-Mediated Sonic Hedgehog Signaling Is Affected in adamts5 Morphants
h
h
d h
d
d A A
ld l
d
l
d
l
2.5. Receptor-Mediated Sonic Hedgehog Signaling Is Affected in adamts5 Morphants We hypothesized that reduced ADAMTS5 could lead to an altered extracellular environment
that might disrupt Shh signaling, and that since adaxial mesoderm is in closer proximity to the
notochord it might be less disrupted compared to the paraxial mesoderm. Therefore, cyclopamine,
an antagonist of Smoothened (Smo), a receptor in the Shh signaling pathway [28] was used to
understand whether Shh signaling through Smo was impaired in adamts5 morphants. The presence
of 5 μM cyclopamine did not affect adaxial myod expression at 12 hpf in wild-type embryos (Figure
4A(g–I),B). However, treatment of adamts5 morphants with 5 μM cyclopamine severely affected
We hypothesized that reduced ADAMTS5 could lead to an altered extracellular environment that
might disrupt Shh signaling, and that since adaxial mesoderm is in closer proximity to the notochord
it might be less disrupted compared to the paraxial mesoderm. Therefore, cyclopamine, an antagonist
of Smoothened (Smo), a receptor in the Shh signaling pathway [28] was used to understand whether
Shh signaling through Smo was impaired in adamts5 morphants. The presence of 5 µM cyclopamine
did not affect adaxial myod expression at 12 hpf in wild-type embryos (Figure 4A(g–I),B). However, Int. J. Mol. Sci. 2018, 19, 766 6 of 12 treatment of adamts5 morphants with 5 µM cyclopamine severely affected adaxial expression of myod
(Figure 4A(j–l),B) compared to untreated adamts5 morphant embryos (Figure 4A(d–f),B). In a reciprocal
experiment, the Smo agonist, SAG, was used to confirm the dependency of Shh signaling on adamts5
expression. Administration of SAG on wild-type embryos disrupted paraxial myod expression in a
similar manner to adamts5 morphants (Figure 5A(d–I),B). However, the same concentration of SAG
partially rescued the loss of paraxial myod patterning in the adamts5 morphants (Figure 5A(g–l),B). These experiments collectively suggest an interaction between ADAMTS5 and Shh, such that they act
synergistically to stimulate myod expression in adaxial mesoderm (Figure 6). Int. J. Mol. Sci. 2018, 19, x FOR PEER REVIEW
6 of 12
adaxial expression of myod (Figure 4A(j–l),B) compared to untreated adamts5 morphant embryos
(Figure 4A(d–f),B). In a reciprocal experiment, the Smo agonist, SAG, was used to confirm the
dependency of Shh signaling on adamts5 expression. Administration of SAG on wild-type embryos
disrupted paraxial myod expression in a similar manner to adamts5 morphants (Figure 5A(d–I),B). However, the same concentration of SAG partially rescued the loss of paraxial myod patterning in the
adamts5 morphants (Figure 5A(g–l),B). 2.5. Receptor-Mediated Sonic Hedgehog Signaling Is Affected in adamts5 Morphants
h
h
d h
d
d A A
ld l
d
l
d
l
2.5. Receptor-Mediated Sonic Hedgehog Signaling Is Affected in adamts5 Morphants Scale bar = 200 μm; (B) Quantitation of embryos showing present
or absent myod patterning represented in (A) Figure 5. Combinatorial activation of Shh signaling and inhibition of adamts5 rescues paraxial myod
expression. (A) Adaxial and paraxial myod expression in 12 hpf embryos treated with vehicle control
(a–c, arrows), 10 μM Smoothened agonist (Smo agon.) (d–f, arrow/open arrowhead represent
present/absent myod expression in paraxial mesoderm), vehicle + adamts5-MO (g–i, arrow/open
arrowhead represents present/absent myod expression in paraxial mesoderm) and 10 μM Smo agon. + adamts5-MO (j–l, arrows indicate myod expression present in paraxial mesoderm). Scale bar = 100
μm; (B) Quantitation of embryos showing present or absent myod patterning represented in (A). Figure 5. Combinatorial activation of Shh signaling and inhibition of adamts5 rescues paraxial myod
expression. (A) Adaxial and paraxial myod expression in 12 hpf embryos treated with vehicle control
(a–c, arrows), 10 μM Smoothened agonist (Smo agon.) (d–f, arrow/open arrowhead represent
present/absent myod expression in paraxial mesoderm), vehicle + adamts5-MO (g–i, arrow/open
arrowhead represents present/absent myod expression in paraxial mesoderm) and 10 μM Smo agon. + adamts5-MO (j–l, arrows indicate myod expression present in paraxial mesoderm). Scale bar = 100
μm; (B) Quantitation of embryos showing present or absent myod patterning represented in (A). Figure 5. Combinatorial activation of Shh signaling and inhibition of adamts5 rescues paraxial myod
expression. (A) Adaxial and paraxial myod expression in 12 hpf embryos treated with vehicle control
(a–c, arrows), 10 µM Smoothened agonist (Smo agon.)
(d–f, arrow/open arrowhead represent
present/absent myod expression in paraxial mesoderm), vehicle + adamts5-MO (g–i, arrow/open
arrowhead represents present/absent myod expression in paraxial mesoderm) and 10 µM Smo agon. + adamts5-MO (j–l, arrows indicate myod expression present in paraxial mesoderm). Scale bar = 100 µm;
(B) Quantitation of embryos showing present or absent myod patterning represented in (A). Combinatorial activation of Shh signaling and inhibition of adamts5 rescues par mbinatorial activation of Shh signaling expression. (A) Adaxial and paraxial myod expression in 12 hpf embryos treated with vehicle control
(a–c, arrows), 10 μM Smoothened agonist (Smo agon.) (d–f, arrow/open arrowhead represent
present/absent myod expression in paraxial mesoderm), vehicle + adamts5-MO (g–i, arrow/open
arrowhead represents present/absent myod expression in paraxial mesoderm) and 10 μM Smo agon. + adamts5-MO (j–l, arrows indicate myod expression present in paraxial mesoderm). Scale bar = 100
μm; (B) Quantitation of embryos showing present or absent myod patterning represented in (A). Figure 5. 3. Discussion
3. Discussion Zebrafish myogenesis is controlled by multiple pathways [29–31]. Signaling from the notochord
specifies slow-twitch muscle precursors in the adaxial mesoderm [24], which migrate laterally after
somite formation to the most lateral muscle layer [32]. Myotomes develop following elongation and
fusion of somitic cells and their attachment to the somite boundary, with the boundaries between
myotome forming the critical myotendenous junctions that are the primary sites of force generation
[33]. ECM–cell adhesion has been shown to be essential for multiple steps in this process [34]. This
study has identified a novel non-catalytic function of the ECM protein ADAMTS5 in regulating Sonic
Hedgehog signaling that impacted on somite differentiation, with reduced expression of myod in the
paraxial mesoderm and disrupted myotome boundaries. Zebrafish myogenesis is controlled by multiple pathways [29–31]. Signaling from the notochord
specifies slow-twitch muscle precursors in the adaxial mesoderm [24], which migrate laterally after
somite formation to the most lateral muscle layer [32]. Myotomes develop following elongation and
fusion of somitic cells and their attachment to the somite boundary, with the boundaries between
myotome forming the critical myotendenous junctions that are the primary sites of force generation [33]. ECM–cell adhesion has been shown to be essential for multiple steps in this process [34]. This study has
identified a novel non-catalytic function of the ECM protein ADAMTS5 in regulating Sonic Hedgehog
signaling that impacted on somite differentiation, with reduced expression of myod in the paraxial
mesoderm and disrupted myotome boundaries. The phenotypes induced by the adamts5 morphants were rescued with mRNA encoding both
wild-type and catalytically-inactive ADAMTS5. Although unexpected, there is some precedence for
ADAMTS family members demonstrating non-catalytic functions as reviewed recently [35]. For
example, ADAMTS1 has been shown to bind to VEGF through its C-terminal thrombospondin
repeats and spacer domain to block VEGFR2 activation [5]. Moreover, both wild-type and
catalytically-inactive (E363A) ADAMTS15 were able to reduce breast cancer cell migration on matrices
of fibronectin or laminin [6]. Furthermore, enzymatic activity was not required for enhancement of
neurite outgrowth by ADAMTS4, which was instead dependent upon MAP kinase cascade activation
[7]. ADAMTSL family members, which are structurally similar to ADAMTS family members but lack
the N-terminal propeptide and catalytic domain, may also offer some important insights into non-
catalytic functions of ADAMTS family members. 3. Discussion
3. Discussion Most notably, mutations of human ADAMTSL2
have been causally linked to the musculoskeletal disorder Geleophysic Dysplasia [36], where patients
present with severe short stature, joint immobility and cardiac valvular abnormalities. Collectively,
this suggests a role of ADAMTS5 in zebrafish muscle development is likely not related to its
enzymatic function. However, mouse studies have highlighted considerable redundancy amongst
ADAMTS members [35] suggesting that combinatorial targeting might be required to identify
additional functions that may be dependent on enzymatic activity. The phenotypes induced by the adamts5 morphants were rescued with mRNA encoding both
wild-type and catalytically-inactive ADAMTS5. Although unexpected, there is some precedence
for ADAMTS family members demonstrating non-catalytic functions as reviewed recently [35]. For texample, ADAMTS1 has been shown to bind to VEGF through its C-terminal thrombospondin
repeats and spacer domain to block VEGFR2 activation [5]. Moreover, both wild-type and
catalytically-inactive (E363A) ADAMTS15 were able to reduce breast cancer cell migration on matrices
of fibronectin or laminin [6]. Furthermore, enzymatic activity was not required for enhancement
of neurite outgrowth by ADAMTS4, which was instead dependent upon MAP kinase cascade
activation [7]. ADAMTSL family members, which are structurally similar to ADAMTS family members
but lack the N-terminal propeptide and catalytic domain, may also offer some important insights into
non-catalytic functions of ADAMTS family members. Most notably, mutations of human ADAMTSL2
have been causally linked to the musculoskeletal disorder Geleophysic Dysplasia [36], where patients
present with severe short stature, joint immobility and cardiac valvular abnormalities. Collectively,
this suggests a role of ADAMTS5 in zebrafish muscle development is likely not related to its enzymatic
function. However, mouse studies have highlighted considerable redundancy amongst ADAMTS
members [35] suggesting that combinatorial targeting might be required to identify additional functions
that may be dependent on enzymatic activity. Shh is an important regulator of musculoskeletal development, given its role in somite and
neural tube patterning. Duplication, and presumed overexpression, of Shh is associated with
congenital muscular hypertrophy in humans [37]. Shh also enables the formation of the cranial
musculature [38] and polarizes the limb during early morphogenesis [39,40]. Shh has also been
demonstrated to mediate the patterning of somites [41,42]. Shh has the ability to activate myogenesis
in vitro and in vivo [43] with expression and secretion of Shh from the notochord able to induce slow
muscle fiber formation in vivo via myod [24]. 2.5. Receptor-Mediated Sonic Hedgehog Signaling Is Affected in adamts5 Morphants
h
h
d h
d
d A A
ld l
d
l
d
l
2.5. Receptor-Mediated Sonic Hedgehog Signaling Is Affected in adamts5 Morphants Combinatorial activation of Shh signaling and inhibition of adamts5 rescues paraxial myod
expression. (A) Adaxial and paraxial myod expression in 12 hpf embryos treated with vehicle control
(a–c, arrows), 10 μM Smoothened agonist (Smo agon.) (d–f, arrow/open arrowhead represent
present/absent myod expression in paraxial mesoderm), vehicle + adamts5-MO (g–i, arrow/open
arrowhead represents present/absent myod expression in paraxial mesoderm) and 10 μM Smo agon. + adamts5-MO (j–l, arrows indicate myod expression present in paraxial mesoderm). Scale bar = 100
μm; (B) Quantitation of embryos showing present or absent myod patterning represented in (A). Figure 5. Combinatorial activation of Shh signaling and inhibition of adamts5 rescues paraxial myod
expression. (A) Adaxial and paraxial myod expression in 12 hpf embryos treated with vehicle control
(a–c, arrows), 10 µM Smoothened agonist (Smo agon.)
(d–f, arrow/open arrowhead represent
present/absent myod expression in paraxial mesoderm), vehicle + adamts5-MO (g–i, arrow/open
arrowhead represents present/absent myod expression in paraxial mesoderm) and 10 µM Smo agon. + adamts5-MO (j–l, arrows indicate myod expression present in paraxial mesoderm). Scale bar = 100 µm;
(B) Quantitation of embryos showing present or absent myod patterning represented in (A). 7 of 12
7 of 12 Int. J. Mol. Sci. 2018, 19, 766
Int J Mol Sci 2018 19 x FOR Figure 6. Model of interaction between Shh and ADAMTS5. Hypothetical model showing ADAMTS5
and Shh expression synergistically regulate downstream activation of MyoD in adaxial and paraxial
mesoderm. Figure 6. Model of interaction between Shh and ADAMTS5. Hypothetical model showing
ADAMTS5 and Shh expression synergistically regulate downstream activation of MyoD in adaxial and
paraxial mesoderm. Figure 6. Model of interaction between Shh and ADAMTS5. Hypothetical model showing ADAMTS5
and Shh expression synergistically regulate downstream activation of MyoD in adaxial and paraxial
mesoderm. Figure 6. Model of interaction between Shh and ADAMTS5. Hypothetical model showing
ADAMTS5 and Shh expression synergistically regulate downstream activation of MyoD in adaxial and
paraxial mesoderm. 3. Discussion
3. Discussion The adamts5 morphants displayed altered myod
expression in the paraxial—but not adaxial—mesoderm despite levels of shh expression in the
t
h
d b i
ff
t d Thi
i ht b
l i
d b
d
d bi
il bilit
f Shh i th
b
Shh is an important regulator of musculoskeletal development, given its role in somite and neural
tube patterning. Duplication, and presumed overexpression, of Shh is associated with congenital
muscular hypertrophy in humans [37]. Shh also enables the formation of the cranial musculature [38]
and polarizes the limb during early morphogenesis [39,40]. Shh has also been demonstrated to mediate
the patterning of somites [41,42]. Shh has the ability to activate myogenesis in vitro and in vivo [43]
with expression and secretion of Shh from the notochord able to induce slow muscle fiber formation
in vivo via myod [24]. The adamts5 morphants displayed altered myod expression in the paraxial—but
not adaxial—mesoderm despite levels of shh expression in the notochord being unaffected. This might Int. J. Mol. Sci. 2018, 19, 766 8 of 12 be explained by reduced bioavailability of Shh in the absence of ADAMTS5. Since the adaxial
myod-positive cells represent the slow muscle precursors that subsequently move through the fast
muscle region where they impact on fast muscle differentiation [44], it would be of interest to examine
the relative distribution of slow and fast twitch muscle fibers in the adamts5 morphants. The results obtained using agonists and antagonists of the downstream Smo pathway suggest that
ADAMTS may work both upstream, as well as in parallel with Shh signals. Wnt/β-catenin signaling
has been shown to act in co-operation with Shh (and BMPs) in embryonic myogenesis [31]. This could
be mediated, at least partially, via ADAMTS5 since Wnt/β-catenin has been shown to act upstream
of ADAMTS5 in other developmental situations, such as chondrocyte maturation and function [45]. Similarly, defects in Delta/Notch can affect somite boundary formation [46], with this pathway also
shown to induce ADAMTS5 in joint cartilage, providing another potential upstream regulator of
ADAMTS5 during somite differentiation. Defective notochords have been identified in mutants of ECM components, such as fibrillin [47],
collagen [48], the basement membrane proteins laminin alpha [49], beta and gamma [50], as well
cell-associated molecules such as integrins [46]. A number of these defects are due to disrupted
morphogenesis that results from perturbed ECM-cell interactions [49]. 4.1. Zebrafish Lines and Maintenance Wild-type and Tg(acta1:lifeact-GFP)/Tg(acta1:mCherryCaax) [26] zebrafish were maintained, raised
and staged according to standard protocols [54]. Embryos were obtained by mating trios or using a
mass embryo production system (MEPS) (Aquatic Habitats) and raised at 28.5 ◦C. Experiments were
approved by the Deakin University Animal Ethics Committees (G14/2013, 15/05/2013). 3. Discussion
3. Discussion This suggests that altered
morphogenesis as well as disrupted patterning may contribute to the perturbed notochord in adamts5
morphants. In addition, U-shaped myotome boundaries have also been observed in mutants of ECM
components, such as fibronectin [51] and laminin [52], or the alternative ECM processing enzyme
MMP-11 [53], providing precedence for ADAMTS5 impacting on the myotome boundary. This study has identified a new function for the metzincin ADAMTS5. By exploring the role of
ADAMTS5 in zebrafish, understanding has been gained of a potential non-catalytic function in the
regulation of muscle development and maintenance via interaction with the Sonic Hedgehog signaling
pathway. Since both ADAMTS5 and Shh have independent—as well as potential combinatorial—roles
during musculoskeletal development, the complex interplay between ADAMTS and Shh could be
relevant to the development of musculoskeletal diseases, such as muscular dystrophies and arthritis. Further biochemical and functional characterization of potential interactions between ADAMTS5 and
Shh in such diseases may reveal new insights into the development and progression of these diseases. Given that treatment options for these diseases are limited, this knowledge could then be applied to the
development of novel therapeutics that specifically modulate this interaction to slow the progression
of these debilitating conditions. 4.2. Embryo Microinjection and Other Treatments Morpholino antisense oligonucleotides (MOs; Gene Tools) targeting the ATG start codon (5′-
atgctgtcgaaattacaggagtttggcgcgtat) and exon 2/3 splice site (5′-ctatcattgaggacgacggcctgcacgctg
ccttcactgtggctcatgagatc) of zebrafish adamts5 (GenBank: JF778846.1) were used to ablate the adamts5
gene. MOs were solubilized in 1× Danieau buffer and 1 nL injected at a concentration of
1 mg/mL into one-cell stage embryos, as previously described [55]. Alternatively-spliced adamts5
species were confirmed by Sanger sequencing (Australian Genome Research Facility, Melbourne,
Australia) of RT-PCR amplicons generated with flanking primers (5′-ggcggatgtaggaactgtgt and
5′-ttacgcacctcacactgctc). Capped RNA encoding full-length wild-type or catalytically-inactive (E411A) Int. J. Mol. Sci. 2018, 19, 766 9 of 12 ADAMTS5 [21] were synthesized using the T7 mMessage mMachine kit (Thermofisher Scientific,
Scoresby, Victoria, Australia) and 40 pg was microinjected into one-cell stage embryos. For other studies, injected embryos were treated with 5 µM cyclopamine (Sigma-Aldrich, St. Louis,
MO, USA) in DMSO or 10 µM Smoothened agonist SAG (CAS 364590-63-6) (Merck Millipore,
Darmstadt, Germany) in water, along with the corresponding vehicle control at 5.5 hpf and fixed at
12 hpf in 4% PFA/PBS. Supplementary Materials: Supplementary materials can be found at http://www.mdpi.com/1422-0067/19
766/s1. Acknowledgments: The authors thank Thomas Hall and Peter Currie for supplying cDNA constructs and
Christopher Kintakas for gross morphological imaging of adamts5 morphants. Additional thanks go to Suneel
Apte and Sumeda Nandadasa for their helpful discussions regarding this work. Author Contributions: Carolyn M. Dancevic and Daniel R. McCulloch conceived and designed the experiments;
Carolyn M. Dancevic, Yann Gibert, Joachim Berger, Adam D. Smith, Clifford Liongue and Nicole Stupka performed
the experiments; Carolyn M. Dancevic, Yann Gibert, Joachim Berger, Alister C. Ward and Daniel R. McCulloch
analyzed the data; Carolyn M. Dancevic, Alister C. Ward and Daniel R. McCulloch wrote the paper. Conflicts of Interest: The authors declare no conflict of interest. Conflicts of Interest: The authors declare no conflict of interest. 4.3. Whole-Mount In Situ Hybridization and Immunofluorescence Whole-mount in situ hybridization was performed as described [56]. The following antisense
digoxigenin-labelled mRNA probes were synthesized by in vitro transcription: shha and myod [57],
and no-tail (ntl) [58]. Immunofluorescence performed on whole embryos with polyclonal rabbit
anti-propeptide ADAMTS5 (Cat# ab39203-100, Abcam, Pak Shak Kok, New Territories, Hong Kong,
China) at 1:200 followed by anti-rabbit Alexa fluor 594 secondary antibody at 1:500 (Life Technologies,
Carlsbad, CA, USA). Histochemical methods were performed as previously described [59]. 4.4. Western Blotting Cell lysates were extracted from 24 hpf embryos and subjected to Western blotting as described
previously [60], using the polyclonal rabbit antibody against propeptide ADAMTS5 described above
at 1:5000 followed by an anti-rabbit HRP antibody (Cell Signaling Technologies, Danvers, MA, USA) at
1:10,000. Protein concentrations were measured using the Bradford assay and equal loading of protein
confirmed by Coomassie blue staining on duplicate SDS-PAGE gels. References 1. Silver, D.L.; Hou, L.; Somerville, R.; Young, M.E.; Apte, S.S.; Pavan, W.J. The secreted metalloprotease
ADAMTS20 is required for melanoblast survival. PLoS Genet. 2008, 4, e1000003. [CrossRef] [PubMed]
2. Enomoto, H.; Nelson, C.M.; Somerville, R.P.; Mielke, K.; Dixon, L.J.; Powell, K.; Apte, S.S. Cooperation of two
ADAMTS metalloproteases in closure of the mouse palate identifies a requirement for versican proteolysis
in regulating palatal mesenchyme proliferation. Development 2010, 137, 4029–4038. [CrossRef] [PubMed]
3. Jonsson-Rylander, A.C.; Nilsson, T.; Fritsche-Danielson, R.; Hammarstrom, A.; Behrendt, M.; Andersson, J.O.;
Lindgren, K.; Andersson, A.K.; Wallbrandt, P.; Rosengren, B.; et al. Role of ADAMTS-1 in atherosclerosis:
Remodeling of carotid artery, immunohistochemistry, and proteolysis of versican. Arterioscler. Thromb. Vasc. Biol. 2005, 25, 180–185. [PubMed]
4. Porter, S.; Span, P.N.; Sweep, F.C.; Tjan-Heijnen, V.C.; Pennington, C.J.; Pedersen, T.X.; Johnsen, M.;
Lund, L.R.; Romer, J.; Edwards, D.R. ADAMTS8 and ADAMTS15 expression predicts survival in human
breast carcinoma. Int. J. Cancer 2006, 118, 1241–1247. [CrossRef] [PubMed] 1. Silver, D.L.; Hou, L.; Somerville, R.; Young, M.E.; Apte, S.S.; Pavan, W.J. The secreted metalloprotease
ADAMTS20 is required for melanoblast survival. PLoS Genet. 2008, 4, e1000003. [CrossRef] [PubMed] 1. Silver, D.L.; Hou, L.; Somerville, R.; Young, M.E.; Apte, S.S.; Pavan, W.J. The secreted metalloprotease
ADAMTS20 is required for melanoblast survival. PLoS Genet. 2008, 4, e1000003. [CrossRef] [PubMed] 2. Enomoto, H.; Nelson, C.M.; Somerville, R.P.; Mielke, K.; Dixon, L.J.; Powell, K.; Apte, S.S. Cooperation of two
ADAMTS metalloproteases in closure of the mouse palate identifies a requirement for versican proteolysis
in regulating palatal mesenchyme proliferation. Development 2010, 137, 4029–4038. [CrossRef] [PubMed] 2. Enomoto, H.; Nelson, C.M.; Somerville, R.P.; Mielke, K.; Dixon, L.J.; Powell, K.; Apte, S.S. Cooperation of two
ADAMTS metalloproteases in closure of the mouse palate identifies a requirement for versican proteolysis
in regulating palatal mesenchyme proliferation. Development 2010, 137, 4029–4038. [CrossRef] [PubMed] g
g p
y
p
p
3. Jonsson-Rylander, A.C.; Nilsson, T.; Fritsche-Danielson, R.; Hammarstrom, A.; Behrendt, M.; Andersson, J.O.;
Lindgren, K.; Andersson, A.K.; Wallbrandt, P.; Rosengren, B.; et al. Role of ADAMTS-1 in atherosclerosis:
Remodeling of carotid artery, immunohistochemistry, and proteolysis of versican. Arterioscler. Thromb. Vasc. Biol. 2005, 25, 180–185. [PubMed] 3. Jonsson-Rylander, A.C.; Nilsson, T.; Fritsche-Danielson, R.; Hammarstrom, A.; Behrendt, M.; Andersson, J.O.;
Lindgren, K.; Andersson, A.K.; Wallbrandt, P.; Rosengren, B.; et al. Role of ADAMTS-1 in atherosclerosis:
Remodeling of carotid artery, immunohistochemistry, and proteolysis of versican. Arterioscler. Thromb. Vasc. Biol. 2005, 25, 180–185. [PubMed] 4. 4.
Porter, S.; Span, P.N.; Sweep, F.C.; Tjan-Heijnen, V.C.; Pennington, C.J.; Pedersen, T.X.; Johnsen, M.;
Lund, L.R.; Romer, J.; Edwards, D.R. ADAMTS8 and ADAMTS15 expression predicts survival in human
breast carcinoma. Int. J. Cancer 2006, 118, 1241–1247. [CrossRef] [PubMed] 4.5. Statistics Two-tailed paired t-tests were performed between all treatment groups compared to the respective
control groups with scoring performed blind. Significance was achieved at a p-value ≤0.05, with
Gaussian distribution assumed in all cases. Supplementary Materials: Supplementary materials can be found at http://www.mdpi.com/1422-0067/19/3/
766/s1. Supplementary Materials: Supplementary materials can be found at http://www.mdpi.com/1422-0067/19/3/
766/s1 References Glasson, S.S.; Askew, R.; Sheppard, B.; Carito, B.; Blanchet, T.; Ma, H.L.; Flannery, C.R.; Peluso, D.;
Kanki, K.; Yang, Z.; et al. Deletion of active ADAMTS5 prevents cartilage degradation in a murine model of
osteoarthritis. Nature 2005, 434, 644–648. [CrossRef] [PubMed] 13. Didangelos, A.; Mayr, U.; Monaco, C.; Mayr, M. Novel role of ADAMTS-5 protein in proteoglycan turnover
and lipoprotein retention in atherosclerosis. J. Biol. Chem. 2012, 287, 19341–19345. [CrossRef] [PubMed] 14. Yayon, A.; Klagsbrun, M.; Esko, J.D.; Leder, P.; Ornitz, D.M. Cell surface, heparin-like molecules are required
for binding of basic fibroblast growth factor to its high affinity receptor. Cell 1991, 64, 841–848. [CrossRef] 14. Yayon, A.; Klagsbrun, M.; Esko, J.D.; Leder, P.; Ornitz, D.M. Cell surface, heparin-like molecules are required
for binding of basic fibroblast growth factor to its high affinity receptor. Cell 1991, 64, 841–848. [CrossRef]
15. Rapraeger, A.C.; Krufka, A.; Olwin, B.B. Requirement of heparan sulfate for bFGF-mediated fibroblast
growth and myoblast differentiation Science 1991 252 1705–1708 [CrossRef] [PubMed] for binding of basic fibroblast growth factor to its high affinity receptor. Cell 1991, 64, 841–848. [CrossRef]
15. Rapraeger, A.C.; Krufka, A.; Olwin, B.B. Requirement of heparan sulfate for bFGF-mediated fibroblast
growth and myoblast differentiation. Science 1991, 252, 1705–1708. [CrossRef] [PubMed] 15. Rapraeger, A.C.; Krufka, A.; Olwin, B.B. Requirement of heparan sulfate for bFGF-mediated fibroblast
growth and myoblast differentiation. Science 1991, 252, 1705–1708. [CrossRef] [PubMed] 16. Witt, R.M.; Hecht, M.L.; Pazyra-Murphy, M.F.; Cohen, S.M.; Noti, C.; van Kuppevelt, T.H.; Fuller, M.;
Chan, J.A.; Hopwood, J.J.; Seeberger, P.H.; et al. Heparan sulfate proteoglycans containing a glypican 5 core
and 2-O-sulfo-iduronic acid function as sonic hedgehog co-receptors to promote proliferation. J. Biol. Chem. 2013, 288, 26275–26288. [CrossRef] [PubMed] 17. Nandadasa, S.; Nelson, C.M.; Apte, S.S. ADAMTS9-mediated extracellular matrix dynamics regulates
umbilical cord vascular smooth muscle differentiation and rotation. Cell Rep. 2015, 11, 1519–1528. [CrossRef]
[PubMed] 18. Lin, A.C.; Seeto, B.L.; Bartoszko, J.M.; Khoury, M.A.; Whetstone, H.; Ho, L.; Hsu, C.; Ali, S.A.; Alman, B.A. Modulating hedgehog signaling can attenuate the severity of osteoarthritis. Nat. Med. 2009, 15, 1421–1425. [CrossRef] [PubMed] 19. Stupka, N.; Kintakas, C.; White, J.D.; Fraser, F.W.; Hanciu, M.; Aramaki-Hattori, N.; Martin, S.; Coles, C.;
Collier, F.; Ward, A.C.; et al. Versican processing by a disintegrin-like and metalloproteinase domain
with thrombospondin-1 repeats proteinases-5 and -15 facilitates myoblast fusion. J. Biol. Chem. 2013, 288,
1907–1917. [CrossRef] [PubMed] 20. References Porter, S.; Span, P.N.; Sweep, F.C.; Tjan-Heijnen, V.C.; Pennington, C.J.; Pedersen, T.X.; Johnsen, M.;
Lund, L.R.; Romer, J.; Edwards, D.R. ADAMTS8 and ADAMTS15 expression predicts survival in human
breast carcinoma. Int. J. Cancer 2006, 118, 1241–1247. [CrossRef] [PubMed] 4. Porter, S.; Span, P.N.; Sweep, F.C.; Tjan-Heijnen, V.C.; Pennington, C.J.; Pedersen, T.X.; Johnsen, M.;
Lund, L.R.; Romer, J.; Edwards, D.R. ADAMTS8 and ADAMTS15 expression predicts survival in human
breast carcinoma. Int. J. Cancer 2006, 118, 1241–1247. [CrossRef] [PubMed] Int. J. Mol. Sci. 2018, 19, 766 10 of 12 5. Iruela-Arispe, M.L.; Carpizo, D.; Luque, A. ADAMTS1: A matrix metalloprotease with angioinhibitory
properties. Ann. N. Y. Acad. Sci. 2003, 995, 183–190. [CrossRef] [PubMed] 5. Iruela-Arispe, M.L.; Carpizo, D.; Luque, A. ADAMTS1: A matrix metalloprotease with angioinhibitory
properties. Ann. N. Y. Acad. Sci. 2003, 995, 183–190. [CrossRef] [PubMed] 6. Kelwick, R.; Wagstaff, L.; Decock, J.; Roghi, C.; Cooley, L.S.; Robinson, S.D.; Arnold, H.; Gavrilovic, J.;
Jaworski, D.M.; Yamamoto, K.; et al. Metalloproteinase-dependent and -independent processes contribute
to inhibition of breast cancer cell migration, angiogenesis and liver metastasis by a disintegrin and
metalloproteinase with thrombospondin motifs-15. Int. J. Cancer 2014, 136, E14–E26. [CrossRef] [PubMed] 7. Hamel, M.G.; Ajmo, J.M.; Leonardo, C.C.; Zuo, F.; Sandy, J.D.; Gottschall, P.E. Multimodal signaling by the
ADAMTSs (a disintegrin and metalloproteinase with thrombospondin motifs) promotes neurite extension. Exp. Neurol. 2008, 210, 428–440. [CrossRef] [PubMed] 8. McCulloch, D.R.; Nelson, C.M.; Dixon, L.J.; Silver, D.L.; Wylie, J.D.; Lindner, V.; Sasaki, T.; Cooley, M.A.;
Argraves, W.S.; Apte, S.S. ADAMTS metalloproteases generate active versican fragments that regulate
interdigital web regression. Dev. Cell 2009, 17, 687–698. [CrossRef] [PubMed] 9. Dubail, J.; Aramaki-Hattori, N.; Bader, H.L.; Nelson, C.M.; Katebi, N.; Matuska, B.; Olsen, B.R.; Apte, S.S. A new Adamts9 conditional mouse allele identifies its non-redundant role in interdigital web regression. Genesis 2014, 52, 702–712. [CrossRef] [PubMed] 10. Dupuis, L.E.; McCulloch, D.R.; McGarity, J.D.; Bahan, A.; Wessels, A.; Weber, D.; Diminich, A.M.;
Nelson, C.M.; Apte, S.S.; Kern, C.B. Altered versican cleavage in ADAMTS5 deficient mice; a novel etiology
of myxomatous valve disease. Dev. Biol. 2011, 357, 152–164. [CrossRef] [PubMed] 11. Stanton, H.; Rogerson, F.M.; East, C.J.; Golub, S.B.; Lawlor, K.E.; Meeker, C.T.; Little, C.B.; Last, K.; Farmer, P.J.;
Campbell, I.K.; et al. ADAMTS5 is the major aggrecanase in mouse cartilage in vivo and in vitro. Nature
2005, 434, 648–652. [CrossRef] [PubMed] 12. References Brunet, F.G.; Fraser, F.W.; Binder, M.J.; Smith, A.D.; Kintakas, C.; Dancevic, C.M.; Ward, A.C.;
McCulloch, D.R. The evolutionary conservation of the A Disintegrin-like and Metalloproteinase domain
with Thrombospondin-1 motif metzincins across vertebrate species and their expression in teleost zebrafish. BMC Evol. Biol. 2015, 15, 22. [CrossRef] [PubMed] 21. Longpre, J.M.; McCulloch, D.R.; Koo, B.H.; Alexander, J.P.; Apte, S.S.; Leduc, R. Characterization of
proADAMTS5 processing by proprotein convertases. Int. J. Biochem. Cell Biol. 2009, 41, 1116–1126. [CrossRef]
[PubMed] Int. J. Mol. Sci. 2018, 19, 766 11 of 12 22. Brunet, F.; Kintakas, C.; Smith, A.D.; McCulloch, D.R. The function of the hyalectan class of proteoglycans
and their binding partners during vertebrate development. In Advances in Medicine and Biology; Nova Science
Publishers Inc.: Hauppauge, NY, USA, 2012; Volume 52, pp. 49–96. 23. Dancevic, C.M. Deakin University, Waurn Ponds, Victoria, Australia. Unpublished data, 2015. 24. Blagden, C.S.; Currie, P.D.; Ingham, P.W.; Hughes, S.M. Notochord induction of zebrafi
mediated by Sonic hedgehog. Genes Dev. 1997, 11, 2163–2175. [CrossRef] [PubMed] 25. Halpern, M.E.; Ho, R.K.; Walker, C.; Kimmel, C.B. Induction of muscle pioneers and floor plate is
distinguished by the zebrafish no tail mutation. Cell 1993, 75, 99–111. [CrossRef] 26. Berger, J.; Tarakci, H.; Berger, S.; Li, M.; Hall, T.E.; Arner, A.; Currie, P.D. Loss of Tropomodulin4 in the
zebrafish mutant trage causes cytoplasmic rod formation and muscle weakness reminiscent of nemaline
myopathy. Dis. Model. Mech. 2014, 7, 1407–1415. [CrossRef] [PubMed] 27. Schauerte, H.E.; van Eeden, F.J.; Fricke, C.; Odenthal, J.; Strahle, U.; Haffter, P. Sonic hedgehog is not required
for the induction of medial floor plate cells in the zebrafish. Development 1998, 125, 2983–2993. [PubMed] 28. Lewis, C.; Krieg, P.A. Reagents for developmental regulation of Hedgehog signaling. Methods 2014, 66,
390–397. [CrossRef] [PubMed] 29. Stickney, H.L.; Barresi, M.J.; Devoto, S.H. Somite development in zebrafish. Dev. Dyn. 2000, 219, 287–303. [CrossRef] 30. Rida, P.C.; Le Minh, N.; Jiang, Y.J. A Notch feeling of somite segmentation and beyond. Dev. Biol. 2004, 265,
2–22. [CrossRef] [PubMed] 31. Von Maltzahn, J.; Chang, N.C.; Bentzinger, C.F.; Rudnicki, M.A. Wnt signaling in myogenesis. Trends Cell Biol. 2012, 22, 602–609. [CrossRef] [PubMed] 32. Devoto, S.H.; Melancon, E.; Eisen, J.S.; Westerfield, M. Identification of separate slow and fast muscle
precursor cells in vivo, prior to somite formation. Development 1996, 122, 3371–3380. [PubMed] 33. Long, J.H.; Adcock, B.; Root, R.G. Force transmission via axial tendons in undulating fish: A dynamic
analysis. Comp. Biochem. References Physiol. A Mol. Integr. Physiol. 2002, 133, 911–929. [CrossRef] 34. Goody, M.F.; Sher, R.B.; Henry, C.A. Hanging on for the ride: Adhesion to the extracellular matrix mediates
cellular responses in skeletal muscle morphogenesis and disease. Dev. Biol. 2015, 401, 75–91. [CrossRef]
[PubMed] 35. Dancevic, C.M.; McCulloch, D.R.; Ward, A.C. The ADAMTS hyalectanase family: Biological insights from
diverse species. Biochem. J. 2016, 473, 2011–2022. [CrossRef] [PubMed] 36. Le Goff, C.; Morice-Picard, F.; Dagoneau, N.; Wang, L.W.; Perrot, C.; Crow, Y.J.; Bauer, F.; Flori, E.;
Prost-Squarcioni, C.; Krakow, D.; et al. ADAMTSL2 mutations in geleophysic dysplasia demonstrate
a role for ADAMTS-like proteins in TGF-β bioavailability regulation. Nat. Genet. 2008, 40, 1119–1123. [CrossRef] [PubMed] 37. Kroeldrup, L.; Kjaergaard, S.; Kirchhoff, M.; Kock, K.; Brasch-Andersen, C.; Kibaek, M.; Ousager, L.B. Duplication of 7q36.3 encompassing the Sonic Hedgehog (SHH) gene is associated with congenital muscular
hypertrophy. Eur. J. Med. Genet. 2012, 55, 557–560. [CrossRef] [PubMed] 38. Balczerski, B.; Zakaria, S.; Tucker, A.S.; Borycki, A.G.; Koyama, E.; Pacifici, M.; Francis-West, P. Distinct
spatiotemporal roles of hedgehog signalling during chick and mouse cranial base and axial skeleton
development. Dev. Biol. 2012, 371, 203–214. [CrossRef] [PubMed] p
9. Riddle, R.D.; Johnson, R.L.; Laufer, E.; Tabin, C. Sonic hedgehog mediates the polarizing activity of the Z
Cell 1993, 75, 1401–1416. [CrossRef] 40. Chiang, C.; Litingtung, Y.; Harris, M.P.; Simandl, B.K.; Li, Y.; Beachy, P.A.; Fallon, J.F. Manifestation of the
limb prepattern: Limb development in the absence of sonic hedgehog function. Dev. Biol. 2001, 236, 421–435. [CrossRef] [PubMed] 41. Fan, C.M.; Tessier-Lavigne, M. Patterning of mammalian somites by surface ectoderm and notochord:
Evidence for sclerotome induction by a hedgehog homolog. Cell 1994, 79, 1175–1186. [CrossRef] 42. Johnson, R.L.; Laufer, E.; Riddle, R.D.; Tabin, C. Ectopic expression of Sonic hedgehog alters dorsal-ventral
patterning of somites. Cell 1994, 79, 1165–1173. [CrossRef] 43. Duprez, D.; Fournier-Thibault, C.; Le Douarin, N. Sonic Hedgehog induces proliferation of committed
skeletal muscle cells in the chick limb. Development 1998, 125, 495–505. [PubMed] 44. Henry, C.A.; Amacher, S.L. Zebrafish slow muscle cell migration induces a wave of fast muscle
morphogenesis. Dev. Cell 2004, 7, 917–923. [CrossRef] [PubMed] 12 of 12 Int. J. Mol. Sci. 2018, 19, 766 45. Tamamura, Y.; Otani, T.; Kanatani, N.; Koyama, E.; Kitagaki, J.; Komori, T.; Yamada, Y.; Costantini, F.;
Wakisaka, S.; Pacifici, M.; et al. Developmental regulation of Wnt/β-catenin signals is required for growth
plate assembly, cartilage integrity, and endochondral ossification. J. Biol. References Chem. 2005, 280, 19185–19195. [CrossRef] [PubMed] 46. Julich, D.; Geisler, R.; Holley, S.A. Tübingen 2000 Screen Consortium. Integrinalpha5 and delta/notch
signaling have complementary spatiotemporal requirements during zebrafish somitogenesis. Dev. Cell 2005,
8, 575–586. [PubMed] 47. Gansner, J.M.; Madsen, E.C.; Mecham, R.P.; Gitlin, J.D. Essential role for fibrillin-2 in zebrafish notochord
and vascular morphogenesis. Dev. Dyn. 2008, 237, 2844–2861. [CrossRef] [PubMed] 48. Gray, R.S.; Wilm, T.P.; Smith, J.; Bagnat, M.; Dale, R.M.; Topczewski, J.; Johnson, S.L.; Solnica-Krezel, L. Loss
of col8a1a function during zebrafish embryogenesis results in congenital vertebral malformations. Dev. Biol. 2014, 386, 72–85. [CrossRef] [PubMed] 49. Parsons, M.J.; Pollard, S.M.; Saude, L.; Feldman, B.; Coutinho, P.; Hirst, E.M.; Stemple, D.L. Zebrafish
mutants identify an essential role for laminins in notochord formation. Development 2002, 129, 3137–3146. [PubMed] 50. Pollard, S.M.; Parsons, M.J.; Kamei, M.; Kettleborough, R.N.; Thomas, K.A.; Pham, V.N.; Bae, M.K.; Scott, A.;
Weinstein, B.M.; Stemple, D.L. Essential and overlapping roles for laminin alpha chains in notochord and
blood vessel formation. Dev. Biol. 2006, 289, 64–76. [CrossRef] [PubMed] 51. Snow, C.J.; Peterson, M.T.; Khalil, A.; Henry, C.A. Muscle development is disrupted in zebrafish embryos
deficient for fibronectin. Dev. Dyn. 2008, 237, 2542–2553. [CrossRef] [PubMed] 52. Peterson, M.T.; Henry, C.A. Hedgehog signaling and laminin play unique and synergistic roles in muscle
development. Dev. Dyn. 2010, 239, 905–913. [CrossRef] [PubMed] 53. Jenkins, M.H.; Alrowaished, S.S.; Goody, M.F.; Crawford, B.D.; Henry, C.A. Laminin and Matrix
metalloproteinase 11 regulate Fibronectin levels in the zebrafish myotendinous junction. Skelet. Muscle 2016,
6, 18. [CrossRef] [PubMed] 54. Kimmel, C.B.; Ballard, W.W.; Kimmel, S.R.; Ullmann, B.; Schilling, T.F. Stages of embryonic development of
the zebrafish. Dev. Dyn. 1995, 203, 253–310. [CrossRef] [PubMed] 5. Nasevicius, A.; Ekker, S.C. Effective targeted gene ‘knockdown’ in zebrafish. Nat Genet 2000, 26, 216–
[CrossRef] [PubMed] 6. Thisse, C.; Thisse, B. High-resolution in situ hybridization to whole-mount zebrafish embryos. Nat. Pr
2008, 3, 59–69. [CrossRef] [PubMed] 57. Gibert, Y.; Gajewski, A.; Meyer, A.; Begemann, G. Induction and prepatterning of the zebrafish pectoral fin
bud requires axial retinoic acid signaling. Development 2006, 133, 2649–2659. [CrossRef] [PubMed] 58. Begemann, G.; Schilling, T.F.; Rauch, G.J.; Geisler, R.; Ingham, P.W. The zebrafish neckless mutation reveals a
requirement for raldh2 in mesodermal signals that pattern the hindbrain. Development 2001, 128, 3081–3094. [PubMed] 59. Lange, M.; Norton, W.; Coolen, M.; Chaminade, M.; Merker, S.; Proft, F.; Schmitt, A.; Vernier, P.; Lesch, K.P.;
Bally-Cuif, L. © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/). References The ADHD-susceptibility gene lphn3.1 modulates dopaminergic neuron formation and
locomotor activity during zebrafish development. Mol. Psychiatry 2012, 17, 946–954. [CrossRef] [PubMed] 59. Lange, M.; Norton, W.; Coolen, M.; Chaminade, M.; Merker, S.; Proft, F.; Schmitt, A.; Vernier, P.; Lesch, K.P.;
Bally-Cuif, L. The ADHD-susceptibility gene lphn3.1 modulates dopaminergic neuron formation and
locomotor activity during zebrafish development. Mol. Psychiatry 2012, 17, 946–954. [CrossRef] [PubMed]
60. Dancevic, C.M.; Fraser, F.W.; Smith, A.D.; Stupka, N.; Ward, A.C.; McCulloch, D.R. Biosynthesis and
expression of a Disintegrin-like and Metalloproteinase domain with Thrombospondin-1 repeats-15: A novel y
g
p
y
y
60. Dancevic, C.M.; Fraser, F.W.; Smith, A.D.; Stupka, N.; Ward, A.C.; McCulloch, D.R. Biosynthesis and
expression of a Disintegrin-like and Metalloproteinase domain with Thrombospondin-1 repeats-15: A novel
versican-cleaving proteoglycanase. J. Biol. Chem. 2013, 288, 37267–37276. [CrossRef] [PubMed] © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/).
|
https://openalex.org/W2910378202
|
https://europepmc.org/articles/pmc6326506?pdf=render
|
English
| null |
Association of PPARGC1A Gly428Ser (rs8192678) polymorphism with potential for athletic ability and sports performance: A meta-analysis
|
PloS one
| 2,019
|
cc-by
| 10,592
|
RESEARCH ARTICLE Phuntila TharabenjasinID1*, Noel Pabalan1, Hamdi Jarjanazi2 1 Chulabhorn International College of Medicine, Thammasat University, Pathum Thani, Thailand,
2 Environmental Monitoring and Reporting Branch, Ontario Ministry of the Environment Conservation and
Parks, Toronto, Ontario, Canada a1111111111
a1111111111
a1111111111
a1111111111
a1111111111 a1111111111
a1111111111
a1111111111
a1111111111
a1111111111 * pacezen@yahoo.com * pacezen@yahoo.com Methods Multi-database literature search yielded 14 articles (16 studies) for inclusion. Pooled odds
ratios (ORs) and 95% confidence intervals (CI) were used to estimate associations. Sum-
mary effects were modified based on statistical power. Subgroup analysis was based on SP
(power, endurance and mixed) and race (Caucasians and Asians). Heterogeneity was
assessed with the I2 metric and its sources examined with outlier analysis which dichoto-
mized our findings into pre- (PRO) and post-outlier (PSO). Copyright: © 2019 Tharabenjasin et al. This is an
open access article distributed under the terms of
the Creative Commons Attribution License, which
permits unrestricted use, distribution, and
reproduction in any medium, provided the original
author and source are credited. OPEN ACCESS Genetics plays a role in determining potential for athletic ability (AA) and sports performance
(SP). In this study, AA involves comparing sedentary controls with competitive athletes in
power and endurance activities as well as a mix between the two (SP). However, variable
results from genetic association studies warrant a meta-analysis to obtain more precise esti-
mates of the association between PPARGC1A Gly482Ser polymorphism and AA/SP. Citation: Tharabenjasin P, Pabalan N, Jarjanazi H
(2019) Association of PPARGC1A Gly428Ser
(rs8192678) polymorphism with potential for
athletic ability and sports performance: A meta-
analysis. PLoS ONE 14(1): e0200967. https://doi. org/10.1371/journal.pone.0200967 Editor: Joseph Devaney, GeneDx, UNITED STATES
Received: June 24, 2018
Accepted: December 18, 2018
Published: January 9, 2019 Association of PPARGC1A Gly428Ser
(rs8192678) polymorphism with potential for
athletic ability and sports performance: A
meta-analysis Phuntila TharabenjasinID1*, Noel Pabalan1, Hamdi Jarjanazi2 Introduction Athletic ability (AA) determines sports performance (SP). SP is a highly polygenic and com-
plex phenotype as well as having a multifactorial etiology where genetic and environmental
factors contribute to differences among trained athletes [1]. In this study, the role of genetics
in determining AA is examined in the context of strength/power and endurance activities as
well as a mix between the two (SP). The mix represents a continuum of SP activities between
power and endurance. Explosive activities such as sprint and weightlifting characterize
strength/power while endurance involves sustained activities such as marathons and cycling. Muscle strength and sprint activity characterize power phenotypes while that of endurance is
maximal oxygen uptake and economy of movement [2]. Muscle fiber composition in these
two types of athletes differs where activation of types II and I fibers occur during high-intensity
activity in power and endurance performances, respectively [3, 4]. These contrasts stem from
diverging genetic backgrounds of power and endurance athletes that drive their physiology
into different trajectories [5]. Multiple genetic variants are thought to influence muscle function and SP phenotypes [2]. Among the genetic loci associated with SP, peroxisome proliferator-activated receptor gamma
co-activator-1-alpha (PPARGC1A or PGC-1 α) aroused interest for the varied functions of the
proteins it encodes. PPARGC1A is encoded by the gene PPARGC1A in humans which is cru-
cial in training-induced muscle adaptation because it co-activates a span of transcriptional fac-
tors that control myriad biological responses [6]. Studies have shown that several amino acid
polymorphic sites exist within the coding region of PPARGC1A, including Gly482Ser
(rs8192678), which is reported to have functional relevance [7]. Current understanding of Gly482Ser in PPARGC1A is viewed in terms of its impact on
health (e.g. diabetes and obesity) and on athletic phenotype (e.g. endurance sports). In terms
of health impact, physiological evidence has shown that Gly482Ser affects blood lipid levels
and insulin sensitivity. Compared with carriers of 482Gly, those with 482Ser have higher levels
of low density lipoprotein cholesterol [8] and higher insulin resistance [9]. As a result, such
persons have increased risk for Type 2 diabetes [10, 11]. A number of similar and variable find-
ings on the role of Gly482Ser in disease have been reported in the literature that warranted
coverage in published meta-analyses [12–14]. Thus, current knowledge shows that studies on
this polymorphism focused more on health outcomes rather than SP. Conclusion Meta-analytical applications in this study generated evidence that show association
between the Gly allele and AA/SP. These were observed in the overall, Caucasians and sta-
tistically powered comparisons which exhibited consistent significance, stability, robustness,
precision and lack of bias. Our central findings rest on association of the Gly allele with
endurance and power, differentially favoring the latter over the former. Meta-analysis PPARGC1A athletic ability sports performance F, Fixed-effects; Fs, Favoring SP; Gly, Glycine; GS,
Gain in significance; HWE, Hardy-Weinberg
Equilibrium; I2, Measure of variability expressed in
%; K, number designation of the article; Log OR,
Logarithm of standardized OR; Maf, Minor allele
frequency; M-H, Mantel-Haenszel; n, Number of
studies; OR, Odds ratio; P, P-value; Pa, P-value for
association; Pb, P-value for heterogeneity;
PPARGC1A, Peroxisome proliferator-activated
receptor gamma co-activator-1-alpha; PRISMA,
Preferred Reporting Items for Systematic Reviews
and Meta-Analyses; PRO, Pre-outlier; PSO, Post-
outlier; R, Random-effects; RH, Reduced
heterogeneity; RNS, Retained non-significance; RS,
Retained significance; SE, Standard error; Ser,
Serine; SP, Sports performance. Results Gly allele effects significantly favoring AA/SP (OR > 1.0, P < 0.05) form the core of our find-
ings in: (i) homogeneous overall effect at the post-modified, PSO level (OR 1.13, 95% CI
1.03–1.25, P = 0.01, I2 = 0%); (ii) initially homogeneous power SP (ORs 1.22–1.25, 95% CI
1.05–1.44, P = 0.003–0.008, I2 = 0%) which precluded outlier treatment; (iii) PRO Cauca-
sian outcomes (ORs 1.29–1.32, 95% CI 1.12–1.54, P = 0.0005) over that of Asians with a
pooled null effect (OR 0.99, 95% CI 0.72–1.99, P = 0.53–0.92) and (iv) homogeneous all >
80% (ORs 1.19–1.38, 95% CI 1.05–1.66, P = 0.0007–0.007, I2 = 0%) on account of high sta-
tistical power (both study-specific and combined). In contrast, none of the Ser allele effects
significantly favored AA/SP and no Ser-Gly genotype outcome favored AA/SP. The core sig-
nificant outcomes were robust and showed no evidence of publication bias. Data Availability Statement: All relevant data are
within the paper and its Supporting Information
files. Funding: The authors received no specific funding
for this work. Competing interests: The authors have declared
that no competing interests exist. Competing interests: The authors have declared
that no competing interests exist. Abbreviations: A, Asian; AM, Analysis model; C,
Caucasian; CI, Confidence interval; CID, Confidence
interval difference; Ds, Disfavoring SP; EH, Eliminated heterogeneity; ES, Elevated significance; 1 / 18 PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 Introduction For the studies that focus on the association of PPARGC1A with SP, Gly482Ser has been
regarded as a promising genetic polymorphism in determining SP status for both power and
endurance-type athletes [2]. This promise may likely also determine AA. Information on why
Gly482Ser may predispose to AA is mainly derived from studies that focused on differential
genotype frequencies between athletes and controls, with findings that tend more toward
endurance over that of power. This polymorphism was posited to be the genetic factor that
predetermines aerobic capacity [15]. However, discrepancies among the primary study PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 2 / 18 Meta-analysis PPARGC1A athletic ability sports performance outcomes for Gly428Ser in SP hedge on the type of sport they refer to. For instance, the Ser
allele has been found to be less frequent among elite athletes in endurance [15] and power
[16]. Other studies, however, report the Ser allele as useful in power activities [17]. Thus, while
the Ser allele was reported to disfavor endurance activities, the Gly allele was found useful [18,
19]. Regardless of exercise type, the Gly allele is considered important in athletics [19]. The
reported advantage of Gly allele carriers confers value to this polymorphism in any polygenic
athletic profile [20]. We performed this meta-analysis because reported findings on the role of
Gly428Ser in SP have differed and the number of articles was ripe for synthesizing the variable
findings. Methodological problems may explain the variabilities, which include limited statistical
power, unrecognized confounding factors, misleading definition of phenotypes and stratifica-
tion of populations [21]. Thus, Gly482Ser studies in SP have been heterogeneous given the var-
ious research approaches, variable sample sizes and different population profiles that
characterize them. Such heterogeneity then renders the proposed associations to be inconsis-
tently replicated. Clearly, utilizing research methods that synthesize diverging primary study
results is needed which meta-analysis seems most suitable to resolve. Nevertheless, to the best
of our knowledge, this is the first meta-analysis to examine the role of Gly482Ser in determin-
ing AA/SP. In this study, we focus on the genetic role of this polymorphism in AA/SP by using an array
of meta-analytical techniques such as a scale to evaluate quality of the primary literature, tests
of association, outlier and modifier treatments, sensitivity analysis and tests for publication
bias in order to assess the strength of evidence. This in-depth treatment precludes covering
other SP related genes. Selection of studies Three databases (MEDLINE using PubMed, Science Direct and Google Scholar) were
searched for association studies as of June 15, 2018. Terms used were “peroxisome proliferator-
activated receptor-gamma co-activator 1-alpha”, “PPARGC1A”, “PGC1-α”, “rs8192678”,
“sports performance” and “polymorphism” as medical subject heading and text, restricted to
the English language. Additional eligible studies were identified from references cited in the
retrieved articles. Inclusion criteria were: (i) case–control study design evaluating the associa-
tion between PPARGC1A polymorphisms and SP; (ii) studies comply with the Hardy-Wein-
berg Equilibrium (HWE); (iii) sufficient genotype or allele frequency data to allow calculation
of odds ratios (ORs) and 95% confidence intervals (CIs). Exclusion criteria include: (i) studies
that do not involve SP (e.g. PPARGC1A polymorphism effects in pathophysiological condi-
tions such as diabetes or cases were non-athletes); (ii) studies whose genotype or allele frequen-
cies were otherwise unusable / absent or when available but combined with other
polymorphisms, preventing proper data extraction; (iii) in case of duplicates, we chose the
most recent article; (iv) reviews; (v) no controls; (vi) when controls were present, their fre-
quencies deviated from the HWE and (vii) non-human subjects and non-English articles. Introduction Thus, we view the single-polymorphism approach most suitable for
reasons of brevity and clarity of reporting. Cases and controls Cases in the included studies were athletes who have participated in competitions (the type of
competition is bracketed) which the component studies stratified into (i) top-elite or world-
class [World and/or European Championships or Olympic Games]; (ii) sub-elite [National
level] and (iii) non-elite [regional level]. However, presentation of the genotype data for
PPARGC1A did not differentiate between these stratifications. Controls were generally healthy,
sedentary, without competitive sports experience. Quality assessment of the studies We used the Clark-Baudouin scale to assess methodological quality of the included studies
[21]. This scale has criteria (P-values, statistical power, corrections for multiplicity, compara-
tive sample sizes between cases and controls, genotyping methods and HWE) that are found in
the component articles. In this scale, low, moderate and high have scores of < 5, 5–7 and 8,
respectively. Data extraction Two investigators (PT and NP) independently extracted the data and arrived at consensus. Extracted information from each article comprised of the following: first author’s name, publi-
cation year, country of origin and SP type. S1 Table tabulates information on the quantitative
data. Core information here is of two types: (i) because the literature on PPARGC1A and SP 3 / 18 PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 Meta-analysis PPARGC1A athletic ability sports performance compared athletes with sedentary controls, we examine propensity for AA, rather than SP in
itself. However, SP in this study is contextualized in terms of power, endurance or mixed. (ii)
Genotype data was used to conduct the meta-analysis, which precludes extraneous informa-
tion such as environmental data which was either unmentioned or unquantified in the primary
literature. Of note, HWE was assessed using the application in https://ihg.gsf.de/cgi-bin/hw/
hwa1.pl. Authors were contacted in order to obtain more information on incomplete data. PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 Meta-analysis Gly482Ser associations with SP (OR) were estimated for each study. Presence of zero genotype
values warranted application of the Laplace correction which involves adding a pseudo-count
of one to all values of the data set [22] prior to generating the forest plots. We used the allele-
genotype approach to enable comparison with study-specific outcomes. We thus compared
the following for Gly482Ser: (i) Gly allele with Ser-Gly/Ser-Ser genotype; (ii) Ser allele with
Ser-Gly/Gly-Gly genotype and (iii) Gly/Ser genotype with homozygous Gly-Gly and Ser-Ser
genotypes. Comparing effects on the same baseline, we used raw data from genotype frequen-
cies to calculate pooled ORs. Pooled ORs with their accompanying 95% CIs were used to assess
the strength of evidence. These are: (i) magnitudes of effects are higher or lower when the val-
ues are farther from or closer to the OR value of 1.0 (null effect), respectively; (ii) OR values
are significant when P < 0.05 (two-sided); (iii) distance from P < 0.05, where farther from
(e.g. P < 0.0001) and closer to (e.g. P < 0.04) this value indicates stronger and weaker associa-
tion, respectively and (iv) CID (confidence interval difference) results when the lower CI is
subtracted from the upper CI, which indicates precision of effects. High and low CID values
indicate low and high precision, respectively. Heterogeneity between studies was estimated with the χ 2-based Q test [23], explored with
subgroup analysis [23] and quantified with the I2 statistic which measures variability between
studies [24]. The fixed effects model [25] was used when P 0.10 or I2 < 50%, otherwise we
opted for the random effects model [26], signifying presence of heterogeneity. Sources of het-
erogeneity were detected with the Galbraith plot [27] followed by re-analysis. Three features of
this re-analysis are worth noting: (i) outlier treatment dichotomizes the comparisons into pre-
outlier (PRO) and post-outlier (PSO) which are integrated in the design of the summary tables;
(ii) outlier treatment is applied in PRO which assumes the random-effects status and (iii) PSO
outcomes are fixed-effects, where larger studies are accorded more weight [28]. The Bonfer-
roni correction, applied to independent comparisons only, was used to adjust for multiple 4 / 18 PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 Meta-analysis PPARGC1A athletic ability sports performance testing. Search results Fig 1 outlines the study selection process in a flowchart following PRISMA (Preferred Report-
ing Items for Systematic Reviews and Meta-Analyses) guidelines. Initial search resulted in 523
citations, followed by a series of omissions (S1 List) that eventually yielded 14 articles for inclu-
sion [15–20, 31–38]. Of the 14, two articles [17, 19] presented independent data from two pop-
ulations placing the total number of studies to 16 (Table 1). Characteristics of the included studies Table 1 shows that participants in most of the studies were Euro-Slavic with three (Australia,
Japan and Korea) contributing to geographical heterogeneity [18, 20, 38]. Subgroups by sport
type and race comprised of power [16, 17, 19, 32, 35], endurance [15, 16, 19, 20, 31, 32, 34–36,
38] and mixed [16, 18, 19, 33, 37], Caucasian [15–17, 19, 20, 31–33, 35–37] and Asian [18, 34,
38], respectively. Median and range Clark-Baudouin score of 7.0 (5–9) indicates high method-
ological quality of the component studies. S2 Table shows that the meta-analysis is composed
of seven, 11, and six studies in power, endurance, and mixed, respectively. Quantitative fea-
tures include sample sizes, genotype frequencies in cases/controls and minor allele frequencies
(maf) in each of the sport types (S2 Table). The maf means and standard deviations of Cauca-
sians (0.34 ± 0.05) and Asians (0.49 ± 0.05) differed significantly (t = -4.86, P < 0.001). The
checklists for PRISMA and meta-analysis for genetic association detailed features of this meta-
analysis in accordance with the guidelines (S2 and S3 Tables). Meta-analysis Sensitivity analysis, which involves omitting one study at a time and recalculating the
pooled OR, was used to test for robustness of the summary effects. Publication bias was
assessed on comparisons with 10 studies only [29]. Data were analyzed using Review Man-
ager 5.3 (Cochrane Collaboration, Oxford, England), SIGMASTAT 2.03, SIGMAPLOT 11.0
(Systat Software, San Jose, CA) and WINPEPI [30]. Meta-analysis outcomes Table 2, S4 and S5 Tables summarize the meta-analysis outcomes by order of genetic compari-
sons (Gly and Ser alleles and Ser-Gly genotype). Between these three tables, number of pooled
ORs > 1.0 (favoring SP) was most in Gly allele and least in Ser allele and none in Ser-Gly geno-
type. This positions the Gly allele analysis as central to our findings because it presents the
most convincing evidence indicating the favoring of AA/SP. PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 Meta-analysis PPARGC1A athletic ability sports performance Test of association
Test of
heterogeneity
Test of association
Test of
heterogeneity
Effect of outlier treatment
(Fs)
n
OR
95% CI
Pa
SP
Pb
I2 (%)
AM
n
OR
95% CI
Pa
SP Pb
I2 (%)
AM
Significance
Heterogeneity
PRO
PSO
All
16
1.24
1.08–1.41
0.002
Fs
0.0006
62
R
14
1.16
1.06–1.26
0.001
Fs
0.16
27
F
RS
RH
Power
7
1.25
1.08–1.44
0.003
Fs
0.66
0
F
—
——
——
——
—
—
—
—
—
—
Endurance
11
1.24
1.02–1.51
0.03
Fs
0.0005
68
R
8
1.23
1.08–1.42
0.003
Fs
0.10
41
F
ES
RH
Mixed
6
1.06
0.72–1.55
0.78
Fs
10−4
83
R
4
1.07
0.88–1.30
0.49
Fs
0.31
16
F
RNS
RH
Race
Caucasian
13
1.29
1.12–1.49
0.0005
Fs
0.002
61
R
11
1.19
1.08–1.31
0.0004
Fs
0.19
27
F
RS
RH
Asian
3
0.99
0.79–1.24
0.92
Null
0.34
8
F
—
——
——
——
—
—
—
—
—
—
Modified
All
10
1.23
1.06–1.44
0.008
Fs
0.001
67
R
8
1.13
1.03–1.25
0.01
Fs
0.53
0
F
RS
EH
All > 80%
5
1.38
1.14–1.66
0.0007
Fs
0.005
73
R
3
1.19
1.05–1.34
0.007
Fs
0.95
0
F
RS
EH
Power
6
1.22
1.05–1.42
0.008
Fs
0.64
0
F
—
——
——
——
—
—
—
—
—
—
Endurance
7
1.19
0.94–1.51
0.14
Fs
0.001
73
R
6
1.09
0.94–1.25
0.27
Fs
0.15
38
F
RNS
RH
Mixed
4
1.03
0.59–1.78
0.92
Fs
10−5
89
R
2
1.07
0.84–1.37
0.56
Fs
0.13
57
F
RNS
RH
Race
Caucasian
8
1.32
1.13–1.54
0.0005
Fs
0.009
63
R
6
1.18
1.06–1.32
0.003
Fs
0.78
0
F
RS
EH
Asian
2
0.99
0.72–1.99
0.53
Null
0.46
0
F
—
——
——
——
—
—
—
—
—
—
n: number of studies; Modified: 248 sample size in either case or control; All > 80%: studies with 248 participants in case and in control; PRO: pre-outlier; PSO:
post outlier; OR: odds ratio; CI: confidence interval; Pa: P-value for association
Pa values that survived the Bonferroni correction; Pb: P-value for heterogeneity; AM: analysis model; R: random-effects; F: fixed-effects; SP: sports performance; Fs:
favor SP; ORs = 0.99–1.01 were considered null; RS: retained significance; RNS: retained non-significance; ES: elevated significance; RH: reduced heterogeneity; EH:
Meta analysis PPARGC1A athletic ability sports performance Table 1. Meta-analysis PPARGC1A athletic ability sports performance Characteristics of the included studies that examined the association of PPARGC1A Gly482Ser polymorphism with sports performance. K
First author
[Reference]
Year
n
Country
Race
SP status
Clark-Baudouin score
1
Ahmetov [31]
2009
1
Russia
C
Endurance
7
2
Eynon [32]
2011
1
Israel
C
Power/Endurance
8
3
Gineviciene
[16]
2011
1
Lithuania
C
Power/Endurance/Mixed
9
4
Gineviciene
[33]
2012
1
Lithuania
C
Mixed
7
5
Gineviciene
[17]
2016
2
Lithuania/Russia
C
Power
8
6
Grealy [20]
2015
1
Australia
C
Endurance
7
7
He [34]
2014
1
China
A
Endurance
5
8
Jin [18]
2014
1
Korea
A
Mixed
7
9
Lucia [15]
2005
1
Spain
C
Endurance
7
10
Maciejewska
[19]
2012
2
Poland/Russia
C
Power/Endurance/Mixed
7
11
Maruszak [35]
2012
1
Poland
C
Power/Endurance
5
12
Muniesa [36]
2010
1
Spain
C
Endurance
7
13
Peplonska [37]
2016
1
Poland
C
Mixed
7
14
Yvert [38]
2016
1
Japan
A
Endurance
8
K: number designation of the article; n: number of studies; C: Caucasian; A: Asian; SP: sports performance
https://doi org/10 1371/journal pone 0200967 t001 Table 2. Outlier and modified effects for Gly allele associations with sports performance. Gly allele effects Table 2 shows the Gly allele associations where 20 (91%) of the 22 comparisons favored AA/SP
(OR >1.0). Of the 20 AA/SP favoring outcomes, 14 (70%) were statistically significant
(P < 0.05). Of the 14 significant outcomes, half survived the Bonferroni correction, five in
PRO and two in PSO. These AA/SP favoring and significant features were observed in the
overall (ORs 1.16–1.24, 95% CI 1.06–1.41, P = 0.001–0.002) and Caucasian subgroup (ORs
1.19–1.29, 95% CI 1.08–1.49, P = 0.0004–0.0005). In contrast to Caucasians, the Asian effects
were null and non-significant (OR 0.99, 95% CI 0.79–1.24, P = 0.92). Fig 2 and Table 2 delineate salient differences between power and endurance where both
effects were significant (P < 0.05). The high significance in power (P = 0.003) survived the
Bonferroni correction, but the moderate significance in endurance (P = 0.03) did not
(Table 2). 5 / 18 PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 Meta-analysis PPARGC1A athletic ability sports performance In terms of pooled effects, that in power (OR 1.25, 95% CI 1.08–1.44) was initially homoge-
neous (I2 = 0%), while that in endurance (OR 1.24, 95% CI 1.02–1.51) was initially
Fig 1. Summary flowchart of literature search. PPARGC1A: Peroxisome proliferator-activated receptor gamma co-activator-1-alpha; SP: sports performance; HWE:
Hardy-Weinberg Equilibrium. https://doi.org/10.1371/journal.pone.0200967.g001 In terms of pooled effects, that in power (OR 1.25, 95% CI 1.08–1.44) was initially homoge-
neous (I2 = 0%), while that in endurance (OR 1.24, 95% CI 1.02–1.51) was initially 6 / 18 PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 Meta-analysis PPARGC1A athletic ability sports performance Test of association
Test of
heterogeneity
Test of association
Test of
heterogeneity
Effect of outlier treatment
(Fs)
n
OR
95% CI
Pa
SP
Pb
I2 (%)
AM
n
OR
95% CI
Pa
SP Pb
I2 (%)
AM
Significance
Heterogeneity
PRO
PSO
All
16
1.24
1.08–1.41
0.002
Fs
0.0006
62
R
14
1.16
1.06–1.26
0.001
Fs
0.16
27
F
RS
RH
Power
7
1.25
1.08–1.44
0.003
Fs
0.66
0
F
—
——
——
——
—
—
—
—
—
—
Endurance
11
1.24
1.02–1.51
0.03
Fs
0.0005
68
R
8
1.23
1.08–1.42
0.003
Fs
0.10
41
F
ES
RH
Mixed
6
1.06
0.72–1.55
0.78
Fs
10−4
83
R
4
1.07
0.88–1.30
0.49
Fs
0.31
16
F
RNS
RH
Race
Caucasian
13
1.29
1.12–1.49
0.0005
Fs
0.002
61
R
11
1.19
1.08–1.31
0.0004
Fs
0.19
27
F
RS
RH
Asian
3
0.99
0.79–1.24
0.92
Null
0.34
8
F
—
——
——
——
—
—
—
—
—
—
Modified
All
10
1.23
1.06–1.44
0.008
Fs
0.001
67
R
8
1.13
1.03–1.25
0.01
Fs
0.53
0
F
RS
EH
All > 80%
5
1.38
1.14–1.66
0.0007
Fs
0.005
73
R
3
1.19
1.05–1.34
0.007
Fs
0.95
0
F
RS
EH
Power
6
1.22
1.05–1.42
0.008
Fs
0.64
0
F
—
——
——
——
—
—
—
—
—
—
Endurance
7
1.19
0.94–1.51
0.14
Fs
0.001
73
R
6
1.09
0.94–1.25
0.27
Fs
0.15
38
F
RNS
RH
Mixed
4
1.03
0.59–1.78
0.92
Fs
10−5
89
R
2
1.07
0.84–1.37
0.56
Fs
0.13
57
F
RNS
RH
Race
Caucasian
8
1.32
1.13–1.54
0.0005
Fs
0.009
63
R
6
1.18
1.06–1.32
0.003
Fs
0.78
0
F
RS
EH
Asian
2
0.99
0.72–1.99
0.53
Null
0.46
0
F
—
——
——
——
—
—
—
—
—
—
n: number of studies; Modified: 248 sample size in either case or control; All > 80%: studies with 248 participants in case and in control; PRO: pre-outlier; PSO:
post outlier; OR: odds ratio; CI: confidence interval; Pa: P-value for association
Pa values that survived the Bonferroni correction; Pb: P-value for heterogeneity; AM: analysis model; R: random-effects; F: fixed-effects; SP: sports performance; Fs:
favor SP; ORs = 0.99–1.01 were considered null; RS: retained significance; RNS: retained non-significance; ES: elevated significance; RH: reduced heterogeneity; EH:
eliminated heterogeneity. Values in bold indicate significant associations that favor SP only. Pa values that survived the Bonferroni correction; Pb: P-value for heterogeneity; AM: analysis model; R: random-effects; F: fixed-effects; SP: sports performance; Fs:
favor SP; ORs = 0.99–1.01 were considered null; RS: retained significance; RNS: retained non-significance; ES: elevated significance; RH: reduced heterogeneity; EH:
eliminated heterogeneity. Values in bold indicate significant associations that favor SP only. Meta-analysis PPARGC1A athletic ability sports performance Meta-analysis PPARGC1A athletic ability sports performance Table 1. Characteristics of the included studies that examined the association of PPARGC1A Gly482Ser polymorphism with sports performance. K
First author
[Reference]
Year
n
Country
Race
SP status
Clark-Baudouin score
1
Ahmetov [31]
2009
1
Russia
C
Endurance
7
2
Eynon [32]
2011
1
Israel
C
Power/Endurance
8
3
Gineviciene
[16]
2011
1
Lithuania
C
Power/Endurance/Mixed
9
4
Gineviciene
[33]
2012
1
Lithuania
C
Mixed
7
5
Gineviciene
[17]
2016
2
Lithuania/Russia
C
Power
8
6
Grealy [20]
2015
1
Australia
C
Endurance
7
7
He [34]
2014
1
China
A
Endurance
5
8
Jin [18]
2014
1
Korea
A
Mixed
7
9
Lucia [15]
2005
1
Spain
C
Endurance
7
10
Maciejewska
[19]
2012
2
Poland/Russia
C
Power/Endurance/Mixed
7
11
Maruszak [35]
2012
1
Poland
C
Power/Endurance
5
12
Muniesa [36]
2010
1
Spain
C
Endurance
7
13
Peplonska [37]
2016
1
Poland
C
Mixed
7
14
Yvert [38]
2016
1
Japan
A
Endurance
8
K: number designation of the article; n: number of studies; C: Caucasian; A: Asian; SP: sports performance
https://doi.org/10.1371/journal.pone.0200967.t001
Table 2. Outlier and modified effects for Gly allele associations with sports performance. Meta-analysis PPARGC1A athletic ability sports performance https://doi.org/10.1371/journal.pone.0200967.t002 fied effects for Gly allele associations with sports performanc Table 2. Outlier and modified effects for Gly allele associations with sports performance. n: number of studies; Modified: 248 sample size in either case or control; All > 80%: studies with 248 participants in case and in control; PRO: pre-outlier; PSO:
post outlier; OR: odds ratio; CI: confidence interval; Pa: P-value for association Pa values that survived the Bonferroni correction; Pb: P-value for heterogeneity; AM: analysis model; R: random-effects; F: fixed-effects; SP: sports performance; Fs:
favor SP; ORs = 0.99–1.01 were considered null; RS: retained significance; RNS: retained non-significance; ES: elevated significance; RH: reduced heterogeneity; EH:
eliminated heterogeneity. Values in bold indicate significant associations that favor SP only. PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 7 / 18 Meta-analysis PPARGC1A athletic ability sports performance https://doi.org/10.1371/journal.pone.0200967.g002 Meta-analysis PPARGC1A athletic ability sports performance Fig 2. Forest plot outcome of PPARGC1A Gly allele effects on SP power in the pre-modifier analysis. Diamond denotes the pooled odds ratio (OR). Squares indicate
the OR in each study, with square sizes directly proportional to the weight contribution (%) of each study. Horizontal lines represent 95% confidence intervals (CI). The
Z test for overall effect indicates significance (P = 0.003). The χ2 test shows absence of heterogeneity (P = 0.66, I2 = 0%). LEGEND: M-H: Mantel-Haenszel; I2: measure
of variability expressed in %. Fig 2. Forest plot outcome of PPARGC1A Gly allele effects on SP power in the pre-modifier analysis. Diamond denotes the pooled odds ratio (OR). Squares indicate
the OR in each study, with square sizes directly proportional to the weight contribution (%) of each study. Horizontal lines represent 95% confidence intervals (CI). The
Z test for overall effect indicates significance (P = 0.003). The χ2 test shows absence of heterogeneity (P = 0.66, I2 = 0%). LEGEND: M-H: Mantel-Haenszel; I2: measure
of variability expressed in %. Fig 2. Forest plot outcome of PPARGC1A Gly allele effects on SP power in the pre-modifier analysis. Diamond denotes the pooled odds ratio (OR). Squares indicate
the OR in each study, with square sizes directly proportional to the weight contribution (%) of each study. Horizontal lines represent 95% confidence intervals (CI). The
Z test for overall effect indicates significance (P = 0.003). The χ2 test shows absence of heterogeneity (P = 0.66, I2 = 0%). LEGEND: M-H: Mantel-Haenszel; I2: measure
of variability expressed in %. heterogeneous (I2 = 68%). Endurance heterogeneity warranted outlier treatment, but power
homogeneity did not. Outlier treatment had multiple effects on a number of parameters: (i) heterogeneity was
reduced (Pheterogeneity 0.10) or eliminated (I2 = 0%); (ii) significance was retained (overall,
Caucasian, all > 80%) and elevated (endurance) and (iii) precision of effects was increased
(reduction of CID values from PRO to PSO). The mechanism of outlier treatment is visualized in Figs 3–5. Fig 3 shows the following fea-
tures for Gly allele endurance PRO: (i) heterogeneous (Pheterogeneity = 0.0005, I2 = 68%); (ii)
moderately significant (OR 1.24, 95% CI 1.02–1.51, P = 0.03) and (iii) CID of 0.49 (CI 1.02–
1.51). In Fig 4, the Galbraith plot identifies three studies as the outliers [20, 31, 38] located
above the +2 and below the -2 confidence limits. Meta-analysis PPARGC1A athletic ability sports performance https://doi.org/10.1371/journal.pone.0200967.g004 https://doi.org/10.1371/journal.pone.0200967.g004 https://doi.org/10.1371/journal.pone.0200967.g004 shows (i) reduced heterogeneity (Pheterogeneity = 0.10, I2 = 41%); (ii) increased significance (OR
1.23, 95% CI 1.08–1.42, P = 0.003) and (iii) increased precision with a reduced CID of 0.34 (CI
1.08–1.42). This operation is numerically summarized in Table 2. Meta-analysis PPARGC1A athletic ability sports performance In Fig 5, the PSO outcome (outliers omitted) Fig 3. Forest plot outcome of PPARGC1A Gly allele effects on SP endurance in the PRO analysis. Diamond denotes the pooled odds ratio (OR). Squares indicate the
OR in each study, with square sizes directly proportional to the weight contribution (%) of each study. Horizontal lines represent 95% confidence intervals (CI). The Z
test for overall effect indicates significance (P = 0.03). The χ 2 test shows presence of heterogeneity (P = 0.0005, I2 = 68%). LEGEND: M-H: Mantel-Haenszel; I2: measure
of variability expressed in %; CID, confidence interval difference. Fig 3. Forest plot outcome of PPARGC1A Gly allele effects on SP endurance in the PRO analysis. Diamond denotes the pooled odds ratio (OR). Squares indicate the
OR in each study, with square sizes directly proportional to the weight contribution (%) of each study. Horizontal lines represent 95% confidence intervals (CI). The Z
test for overall effect indicates significance (P = 0.03). The χ 2 test shows presence of heterogeneity (P = 0.0005, I2 = 68%). LEGEND: M-H: Mantel-Haenszel; I2: measure
of variability expressed in %; CID, confidence interval difference. PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 8 / 18 Fig 4. Galbraith plot analysis of Gly allele effects on SP in endurance identifying the sources of heterogeneity. The three studies that lie above and below the +2 and
—2 confidence limits are the outliers. LEGEND: Log OR: logarithm of standardized odds ratio; SE: standard error. https://doi org/10 1371/journal pone 0200967 g004
Meta-analysis PPARGC1A athletic ability sports performance Meta-analysis PPARGC1A athletic ability sports performance Fig 4. Galbraith plot analysis of Gly allele effects on SP in endurance identifying the sources of heterogeneity. The three studies that lie above and below the +2 and
—2 confidence limits are the outliers. LEGEND: Log OR: logarithm of standardized odds ratio; SE: standard error. Fig 4. Galbraith plot analysis of Gly allele effects on SP in endurance identifying the sources of heterogeneity. The th
—2 confidence limits are the outliers. LEGEND: Log OR: logarithm of standardized odds ratio; SE: standard error. https://doi org/10 1371/journal pone 0200967 g004 Fig 4. Galbraith plot analysis of Gly allele effects on SP in endurance identifying the sources of heterogeneity. The three studies that lie above and below the +2 and
—2 confidence limits are the outliers. LEGEND: Log OR: logarithm of standardized odds ratio; SE: standard error. Meta-analysis PPARGC1A athletic ability sports performance Fig 5. Forest plot outcome of outlier treatment on PPARGC1A Gly allele effects on SP endurance in the PSO analysis. Diamond denotes the pooled odds ratio
(OR). Squares indicate the OR in each study, with square sizes directly proportional to the weight contribution (%) of each study. Horizontal lines represent 95%
confidence intervals (CI). The Z test for overall effect indicates high significance given (P = 0.003). The χ 2 test indicates non-heterogeneity (P = 0.10, I2 = 41%). LEGEND: M-H: Mantel-Haenszel; I2: measure of variability expressed in %; CID, confidence interval difference. Fig 5. Forest plot outcome of outlier treatment on PPARGC1A Gly allele effects on SP endurance in the PSO analysis. Diamond denotes the pooled odds ratio
(OR). Squares indicate the OR in each study, with square sizes directly proportional to the weight contribution (%) of each study. Horizontal lines represent 95%
confidence intervals (CI). The Z test for overall effect indicates high significance given (P = 0.003). The χ 2 test indicates non-heterogeneity (P = 0.10, I2 = 41%). LEGEND: M-H: Mantel-Haenszel; I2: measure of variability expressed in %; CID, confidence interval difference. Fig 5. Forest plot outcome of outlier treatment on PPARGC1A Gly allele effects on SP endurance in the PSO analysis. Diamond denotes the pooled odds ratio
(OR). Squares indicate the OR in each study, with square sizes directly proportional to the weight contribution (%) of each study. Horizontal lines represent 95%
confidence intervals (CI). The Z test for overall effect indicates high significance given (P = 0.003). The χ 2 test indicates non-heterogeneity (P = 0.10, I2 = 41%). LEGEND: M-H: Mantel-Haenszel; I2: measure of variability expressed in %; CID, confidence interval difference. Fig 5. Forest plot outcome of outlier treatment on PPARGC1A Gly allele effects on SP endurance in the PSO analysis. Diamond denotes the pooled odds ratio
(OR). Squares indicate the OR in each study, with square sizes directly proportional to the weight contribution (%) of each study. Horizontal lines represent 95%
confidence intervals (CI). The Z test for overall effect indicates high significance given (P = 0.003). The χ 2 test indicates non-heterogeneity (P = 0.10, I2 = 41%). LEGEND: M-H: Mantel-Haenszel; I2: measure of variability expressed in %; CID, confidence interval difference. https://doi.org/10.1371/journal.pone.0200967.g005 assuming an α level of 5%. Outcomes of all > 80% not only significant (ORs 1.19–1.38, 95% CI
1.05–1.66, P = 0.0007–0.007) but homogeneous (I2 = 0%) at the PSO level. In Table 2, modified analysis further highlighted the differences between power and endur-
ance effects. Modified power outcomes (OR 1.22, 95% CI 1.05–1.42, P = 0.008) reflected the
overall effect indicating consistency of significance for this SP type. In contrast, modified
endurance effect lost significance (OR 1.19, 95% CI 0.94–1.51, P = 0.14) when compared to the
overall outcome. Outlier application to the modified and heterogeneous (I2 = 73%) endurance
pooled effect reduced heterogeneity (I2 = 38%) and retained non-significance (P = 0.27).Other
comparisons (PRO and PSO) reduced and eliminated heterogeneity (Overall, Caucasian and
Asia: I2 = 8–27% to 0%). Ser allele and Ser-Gly genotype effects S4 Table shows the Ser allele associations where three (14%) of the 22 comparisons (PRO and
PSO) favored SP (ORs 1.08–1.13, 95% CI 0.83–1.44). Of the three, none were significant
(P > 0.05). Eighteen of the 22 (82%) comparisons in PRO and PSO disfavored SP (ORs 0.57–
0.95, 95% CI 0.41–1.23) of which, two (11%) were significant (P < 0.05). The remaining com-
parison was the null Asian effect (OR 1.01, 95% CI 0.79–1.31, P = 0.91). S5 Table shows the Ser-Gly genotype associations 13 outcomes of which 11 (85%) disfa-
vored SP (ORs 0.80–0.87, 95% CI 0.69–1.00, P < 10−4–0.06) and two (15%) had null (Asian)
outcomes (ORs 1.00–1.01, 95% CI 0.80–1.26, P = 0.95–1.00). All Ser-Gly genotype compari-
sons had the fixed-effects feature indicating initial non-heterogeneity. Modified effects in Gly comparisons Testing a single nucleotide polymorphism using a case-control design has been calculated to
require a sample size of 248 participants in each group to achieve a statistical power of 80%
[39]. To approximate this level and still achieve enough studies, we selected those with at least
248 participants in either cases or controls for Gly allele comparison only. However, we also
included five studies from four papers [19, 31, 35, 37] with > 248 participants both in cases
and in controls which we termed “all > 80%”. Using the GPower program [40], statistical
powers in each of these three studies were calculated to range between 81.0% and 99.9% PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 9 / 18 https://doi.org/10.1371/journal.pone.0200967.g005 Sensitivity analysis and publication bias Sensitivity analysis was performed using a modified protocol that confined this treatment to
the significant Gly allele findings. Pooled effects that retained (P < 0.05) or lost (P > 0.05) sig-
nificance were considered robust and not robust, respectively. Table 3 identifies the robust and
non-robust comparisons. The most robust comparisons were overall and all > 80% in the
PRO analysis and all Caucasian outcomes. The PRO analysis had seven robust outcomes and
as many interfering studies; PSO had half the number of robust outcomes and one undupli-
cated interfering study [19]. All unstable comparisons are attributed to six studies from five PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 10 / 18 Meta-analysis PPARGC1A athletic ability sports performance Table 3. Sensitivity analysis of Gly allele comparisons with significant outcomes favoring sports performance. Comparison
PRO
PSO
Number of References contributing to non-
robustness
Number of robust
outcomes
Overall
Robust
Robust
0
2
Power
Robust
—-
0
1
Endurance
[15, 19, 31, 32,
36]
Robust
5
1
Caucasian
Robust
Robust
0
2
Modified
Overall
Robust
[19]
1
1
All > 80%
Robust
[19]
1
1
Power
Robust
—-
0
1
Caucasian
Robust
Robust
0
2
Number of References contributing to non-
robustness
5
2
Number of robust outcomes
7
4
PRO: pre-outlier; PSO: post-outlier; Numbers in brackets indicate references that contributed to non-robustness
https://doi.org/10.1371/journal.pone.0200967.t003 Table 3. Sensitivity analysis of Gly allele comparisons with significant outcomes favoring sports performance. PRO: pre-outlier; PSO: post-outlier; Numbers in brackets indicate references that contributed to non-robustness articles [15, 19, 31, 32, 36]. Our sensitivity PRO findings highlight the difference between the
SP types where power outcome was robust and endurance was not. Data (study-specific ORs)
used to test for publication bias was determined to be normally distributed (Kolmogorov-
Smirnov test: P > 0.05). Hence, we used the Egger’s regression asymmetry test only which
showed no evidence of publication bias (Table 4). PRO: pre-outlier; PSO: post-outlier; n: number of studies; non-significant P-values (> 0.05) indicate absence of
evidence of publication bias PRO: pre-outlier; PSO: post-outlier; n: number of studies; non-significant P-values (> 0.05) indicate absence of
evidence of publication bias https://doi.org/10.1371/journal.pone.0200967.t004 Summary of effects In this meta-analysis, we present evidence of; (i) Gly allele outcomes favoring the potential for
AA/SP over that of the Ser allele; (ii) within the Gly allele, Caucasians are affected but not
Asians and (iii) Gly allele favors propensity for AA in power SP more than endurance and not
in mixed sports at all. Of note, strength of the potential for power athletics lies in its homoge-
neity and stability of surviving the Bonferroni correction. (iv) Strength of the Gly allele effect is
shown by the all > 80% outcomes showcasing the statistical power of this modified
comparison. Table 4. Publication bias assessment of Gly allele comparisons with significant outcomes favoring sports
performance. Egger’s regression
asymmetry test
n
Intercept
P-value
All PRO
16
-1.15
0.25
All PSO
13
0.23
0.78
Endurance PRO
11
-0.84
0.52
Caucasian PRO
13
-0.93
0.39
Caucasian PSO
11
0.00
1.00 Table 4. Publication bias assessment of Gly allele comparisons with significant outcomes favoring sports
performance. PRO: pre-outlier; PSO: post-outlier; n: number of studies; non-significant P-values (> 0.05) indicate absence of
evidence of publication bias PRO: pre-outlier; PSO: post-outlier; n: number of studies; non-significant P-values (> 0.05) indicate absence of
evidence of publication bias
h
//d i
/10 13 1/j
l
020096
004 11 / 18 PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 Meta-analysis PPARGC1A athletic ability sports performance Our findings delineated which genetic component of Gly428Ser in the PPARGC1A gene
favored SP (Gly allele) and those that did not (Ser allele and Ser-Gly genotype). Subjecting
these components to meta-analysis treatments (outlier, modified and sensitivity) impacted on
the outputs. For example, the combined application of outlier and modifier treatments unrav-
eled favorable features of the pooled outcomes that included reduced/ eliminated heterogene-
ity and elevated statistical power. Outlier treatment attempts to resolve heterogeneity issues
that are inherent in meta-analysis. Modifier treatment operates through exclusion of under-
powered studies. Underpowered outcomes appear to be common in candidate gene studies
[28] and are prone to the risk of Type 1 error. This risk was addressed by generating a compar-
ison with increased statistical power and correcting for multiple comparisons. Thus, all > 80%
modified analysis was created especially in light of significant results [41] and Bonferroni cor-
rection to minimize the possibility of false-positive outcomes [42]. Both outlier and modifier
treatments raise the levels of evidence presented here and highlight the transparency of our
findings. Summary of effects The main findings of this study center on the Gly allele on account of the following: (i) dif-
ferential effects between power and endurance SP were clarified based on statistical (signifi-
cance and correction) and meta-analytical treatments (modifier and sensitivity) and (ii)
statistical significance were observed in all > 80% and the Caucasian subgroup. The SP favor-
ing Gly allele bearing Caucasians but not Asians may be attributed to the significant difference
in maf between the two races. While the Asian subgroup acquired zero heterogeneity on
account of modifier treatment, the Caucasian subgroup acquired homogeneity on account of
modifier and outlier treatments combined. While homogeneous outcomes in meta-analysis improve the quality of evidence, heteroge-
neous results are unavoidable and must be addressed. A pro-active approach to addressing het-
erogeneity is identifying its sources using outlier treatment to re-analyze the results. Our
application of outlier treatment had far-reaching effects, impacting on significance, heteroge-
neity and precision. However, it did not eliminate heterogeneity for the most part. Reduced
heterogeneity, notwithstanding, our meta-analysis findings, such as those in the endurance
outcomes agree with the physiological evidence [18, 19]. However, our meta-analysis results
favor power more than endurance suggesting that PPARGC1A polymorphism may affect other
physiological parameters related to power performance. Variable pooled outcomes (ORs that skirt the null effect [ORs 0.99–1.01] and none that
indicate favoring SP) observed in mixed sports effectively differs from the SP favoring power
outcomes which seem to reflect the inherent phenotypic heterogeneity of this sport type [43]. Ser allele and Ser-Gly genotype effects were consistent in disfavoring SP, regardless of sport
type and outlier treatment [19]. Favoring SP outcomes are underpinned by a number of
important features: (i) similar repeated effects in the comparisons (consistency); (ii) reduced
PSO heterogeneity (outcomes of outlier treatment); (iii) enhanced PSO significance (endur-
ance); (iv) increased precision (reduced CID values from PRO to PSO) and (v) robustness
(resistance to sensitivity treatment) all of which present strong evidence of Gly428Ser associa-
tions with SP. Genetic and physiological correlates PPARGC1A has multiple physiological roles which include: (i) regulating cellular energy
metabolism; (ii) regulating expression of genes that encode key enzymes involved in fatty acid
oxidation [44] and oxidative phosphorylation [15]; (iii) it promoting glucose metabolism
through upregulation of hepatic gluconeogenic genes [45, 46] and (iv) mediating skeletal mus-
cle fiber type switching. 12 / 18 PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 Meta-analysis PPARGC1A athletic ability sports performance Combined peak force/power and ability to sustain high-intensity efforts for extended peri-
ods during a competition [4] is the process that uses oxidative metabolism [15, 19]. Skeletal
muscle fiber type switching involves transition from glycolytic type IIb to mitochondria-rich
types IIa and I which characterizes SP among power athletes. Mitochondrial amount in the
recruited muscle fibers likely determines maximal sustainable power [47]. Not only has
PPARGC1A been identified as master regulator of mitochondrial biogenesis, but it has also
been shown to regulate proteins involved in angiogenesis and anti-oxidant defense as well as
affect expression of inflammatory markers [19, 48]. The PPARGC1A protein has been shown
to control muscle plasticity and suppress inflammatory response [7]. Acute exercise induces
oxidative stress, mobilizes inflammatory response bolstering higher expression of PPARGC1A
that may facilitate endurance athletes’ SP [49–51]. Expression levels of PPARGC1A have been
shown to be altered in response to physiological stress or increased energy demands elicited by
exercise training [52]. Thus, increase in PPARGC1A mRNA levels and its over-expression cor-
responds with delayed fatigue of the contracting muscle [53, 54] and this increase during exer-
cise [54, 55] enhances skeletal muscle oxidative capacity [32, 53, 56]. Investigators have
examined the role of PPARGC1A mRNA expression in SP where its levels were impacted by
exercise training in both mouse and human skeletal muscle [43]. In mouse muscle, PPARGC1A is required to uphold mitochondrial protein expression which is needed for oxida-
tive phosphorylation and perturbation of this cascade results in diminished exercise capacity
[57]. In humans, Mathai et al. demonstrated that one session of protracted endurance activity
induces elevated transcription and mRNA levels of PPARGC1A [46]. PPARGC1A has been
shown to be expressed at high levels in metabolically active tissues where mitochondria are
abundant and oxidative phosphorylation is operational such as brown adipose tissue, heart,
and skeletal muscle, whereas expression level is low in white adipose tissue, liver, and pancreas
[45, 58]. Genetic and physiological correlates Functionality of the Gly482Ser polymorphism could likely affect mRNA expression
and/or protein levels [32]. Therefore, knowledge of genotype may predict AA/SP [15]. Thus,
PPARGC1A is implicated in promoting gene expression and muscle morphology characteristic
of type I oxidative fibers in skeletal muscle. PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 Strengths and limitations Interpreting our findings here is best done in the context of its limitations and strengths. Limi-
tations of our study include: (i) dominating presence of Slavic Caucasian participants (Russia,
Lithuania). This precludes extrapolation of the findings to other ethnic groups. More studies
are warranted to better represent a wider range of ethnic subgroups, particularly Asian popula-
tions; (ii) we did not examine female effects because of data unavailability. Only one [32] of
the component studies presented gender-discriminating data which was insufficient to per-
form subgroup analysis. Although gender differences are not always clear, genes seem to play a
more prominent role in male than in female strength determination [59]; (iii) most of the
component studies were underpowered; (iv) heterogeneity of the PRO findings; (v) elevated
statistical power through modified treatment was countered by non-robustness in the PSO
analysis of overall and all > 80% and (vi) caution maybe warranted in concluding strong asso-
ciations of the Gly allele in our study, given the possibility that this SP increasing allele may be
in linkage disequilibrium with the true functional allele [60]. On the other hand, the following strengths not only add to the epidemiological, clinical and
statistical homogeneity (hence, combinability) of the studies, but also minimize bias and
underpin the magnitude of associations: (i) the combined sample sizes of the overall and SP
types yielded high statistical power (S1 Table); (ii) screening for studies whose controls devi-
ated from HWE effectively corrected for genotyping errors, which minimizes methodological PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 13 / 18 Meta-analysis PPARGC1A athletic ability sports performance weaknesses [61]; (iii) overall methodological quality (determined by the Clark-Baudouin
score) of the included studies was high; (iv) outlier treatment reduce and eliminated heteroge-
neity. Impact of this treatment on significance and precision is viewed to favor our findings;
(v) the PRO overall and power outcomes withstood the Bonferroni correction minimizing the
possibility of a Type 1 error; (vi) sensitivity treatment conferred robustness to all overall and
Caucasian findings, as well as modified overall and all > 80% in PRO and (vii) absence of evi-
dence of publication bias nullifies the notion that it inflates significant pooled outcomes
against non-significant results [28]. Supporting information pp
g
S1 List. Excluded articles. (DOCX)
S1 Table. Quantitative features. (DOCX)
S2 Table. PRISMA checklist. (DOCX)
S3 Table. Checklist meta-analysis on genetic associations. (DOCX)
S4 Table. Ser allele summary outcomes. (DOCX)
S5 Table. Gly-Ser genotype summary outcomes. (DOCX) Conclusions We should point out that interpreting effects of polymorphisms differ between disease and SP,
besides their respective domains in pathology and in normal phenotype. Disease effects are
viewed in terms of protection (reduced risk) or susceptibility (increased risk) both of which
have equal importance especially when significant. Potential for AA/SP effects on the other
hand, is better contextualized when interpreting outcomes that favor SP (OR > 1.0). Thus, our
reason for de-emphasizing ORs < 1.0 is that these values disfavoring AA/SP do not contribute
to promoting AA/SP. p
g
Highlights of our findings rests on the fact that most studies in this study lacked statistical
power (68.8% in the overall analysis), but when the data are combined using meta-analysis,
clear Gly allele effects are uncovered. We recognize that complexity of athletic potential
involves interactions between genetic and non-genetic factors allowing for the possibility of
environmental involvement in modifying Gly482Ser effects. Gene-gene and gene-environ-
ment interactions have been reported to have roles in associations of PPARGC1A polymor-
phisms with SP [5, 17]. While all but one [32] of the 11 articles mentioned gene-environment
interaction, only two addressed haplotype analysis [19, 31]. Nevertheless, all but two [15, 19]
analyzed polymorphisms in other genes, the most common being angiotensin converting
enzyme and α-actinin in six [16, 17, 20, 31, 33, 36] and four articles [17, 20, 33, 36],
respectively. Including other SP-related genes in our meta-analysis would have been logistically prob-
lematic. Additional well-designed studies (including meta-analyses) exploring other parame-
ters would confirm or modify our results in this study and add to the extant knowledge about
the association of PPARGC1A polymorphism and potential for SP. Author Contributions Conceptualization: Phuntila Tharabenjasin, Noel Pabalan. Data curation: Noel Pabalan, Hamdi Jarjanazi. Formal analysis: Phuntila Tharabenjasin, Noel Pabalan, Hamdi Jarjanazi. Investigation: Phuntila Tharabenjasin, Noel Pabalan, Hamdi Jarjanazi. Methodology: Phuntila Tharabenjasin, Noel Pabalan, Hamdi Jarjanazi. Resources: Phuntila Tharabenjasin, Hamdi Jarjanazi. Software: Phuntila Tharabenjasin, Noel Pabalan, Hamdi Jarjanazi. Supervision: Noel Pabalan. Conceptualization: Phuntila Tharabenjasin, Noel Pabalan. Conceptualization: Phuntila Tharabenjasin, Noel Pabalan. Data curation: Noel Pabalan, Hamdi Jarjanazi. Software: Phuntila Tharabenjasin, Noel Pabalan, Hamdi Jarjanazi. Validation: Phuntila Tharabenjasin, Noel Pabalan, Hamdi Jarjanazi. Validation: Phuntila Tharabenjasin, Noel Pabalan, Hamdi Jarjanazi. Visualization: Noel Pabalan, Hamdi Jarjanazi. Writing – original draft: Phuntila Tharabenjasin, Noel Pabalan, Hamdi Jarjanazi. Writing – review & editing: Phuntila Tharabenjasin, Noel Pabalan, Hamdi Jarjanazi. Acknowledgments We thank Prof. Chumpol Pholpramool for reviewing the final draft. S1 List. Excluded articles.
(DOCX) S1 Table. Quantitative features. (DOCX) S1 Table. Quantitative features. (DOCX) S2 Table. PRISMA checklist. (DOCX) S3 Table. Checklist meta-analysis on genetic associations. (DOCX) S3 Table. Checklist meta-analysis on genetic associations. (DOCX) S4 Table. Ser allele summary outcomes. (DOCX) S5 Table. Gly-Ser genotype summary outcomes. (DOCX) 14 / 18 PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 Meta-analysis PPARGC1A athletic ability sports performance PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 References 1. Myburgh KH. What makes an endurance athlete world-class? Not simply a physiological conundrum. Comparative biochemistry and physiology Part A, Molecular & integrative physiology. 2003; 136
(1):171–90. PMID: 14527639. 2. Calo MC, Vona G. Gene polymorphisms and elite athletic performance. Journal of anthropological sci-
ences = Rivista di antropologia: JASS. 2008; 86:113–31. PMID: 19934471. 3. Steinbacher P, Feichtinger RG, Kedenko L, Kedenko I, Reinhardt S, Schonauer AL, et al. The single
nucleotide polymorphism Gly482Ser in the PGC-1alpha gene impairs exercise-induced slow-twitch
muscle fibre transformation in humans. PloS one. 2015; 10(4):e0123881. https://doi.org/10.1371/
journal.pone.0123881 PMID: 25886402; PubMed Central PMCID: PMC4401702. 4. Storey A, Smith HK. Unique aspects of competitive weightlifting: performance, training and physiology. Sports medicine. 2012; 42(9):769–90. https://doi.org/10.2165/11633000-000000000-00000 PMID:
22873835. 5. Ahmetov II, Egorova ES, Gabdrakhmanova LJ, Fedotovskaya ON. Genes and Athletic Performance:
An Update. Medicine and sport science. 2016; 61:41–54. https://doi.org/10.1159/000445240 PMID:
27287076. 6. Lin J, Handschin C, & Spiegelman BM. Metabolic control through the PGC-1 family of transcription
coactivators. Cell Metabolism. 2005; 1:361–70. https://doi.org/10.1016/j.cmet.2005.05.004 PMID:
16054085 7. Handschin C, Spiegelman BM. The role of exercise and PGC1alpha in inflammation and chronic dis-
ease. Nature. 2008; 454(7203):463–9. https://doi.org/10.1038/nature07206 PMID: 18650917; PubMed
Central PMCID: PMC2587487. 8. Zhang SL, Lu WS, Yan L, Wu MC, Xu MT, Chen LH, et al. Association between peroxisome prolifera-
tor-activated receptor-gamma coactivator-1alpha gene polymorphisms and type 2 diabetes in southern
Chinese population: role of altered interaction with myocyte enhancer factor 2C. Chinese medical jour-
nal. 2007; 120(21):1878–85. PMID: 18067759. 9. Hara K, Tobe K, Okada T, Kadowaki H, Akanuma Y, Ito C, et al. A genetic variation in the PGC-1 gene
could confer insulin resistance and susceptibility to Type II diabetes. Diabetologia. 2002; 45(5):740–3. https://doi.org/10.1007/s00125-002-0803-z PMID: 12107756. 10. Andrulionyte L, Peltola P, Chiasson JL, Laakso M, Group S-NS. Single nucleotide polymorphisms of
PPARD in combination with the Gly482Ser substitution of PGC-1A and the Pro12Ala substitution of PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 15 / 18 Meta-analysis PPARGC1A athletic ability sports performance PPARG2 predict the conversion from impaired glucose tolerance to type 2 diabetes: the STOP-NIDDM
trial. Diabetes. 2006; 55(7):2148–52. https://doi.org/10.2337/db05-1629 PMID: 16804087. PPARG2 predict the conversion from impaired glucose tolerance to type 2 diabetes: the STOP-NIDDM
trial. Diabetes. 2006; 55(7):2148–52. https://doi.org/10.2337/db05-1629 PMID: 16804087. 11. Ek J, Andersen G, Urhammer SA, Gaede PH, Drivsholm T, Borch-Johnsen K, et al. References Mutation analysis
of peroxisome proliferator-activated receptor-gamma coactivator-1 (PGC-1) and relationships of identi-
fied amino acid polymorphisms to Type II diabetes mellitus. Diabetologia. 2001; 44(12):2220–6. https://
doi.org/10.1007/s001250100032 PMID: 11793024. 12. Barroso I, Luan J, Sandhu MS, Franks PW, Crowley V, Schafer AJ, et al. Meta-analysis of the Gly482-
Ser variant in PPARGC1A in type 2 diabetes and related phenotypes. Diabetologia. 2006; 49(3):501–5. https://doi.org/10.1007/s00125-005-0130-2 PMID: 16435105. 13. Sharma R, Matharoo K, Kapoor R, Bhanwer AJS. Association of PGC-1alpha gene with type 2 diabetes
in three unrelated endogamous groups of North-West India (Punjab): a case-control and meta-analysis
study. Molecular genetics and genomics: MGG. 2018; 293(2):317–29. https://doi.org/10.1007/s00438-
017-1385-2 PMID: 29063962. 14. Yang Y, Mo X, Chen S, Lu X, Gu D. Association of peroxisome proliferator-activated receptor gamma
coactivator 1 alpha (PPARGC1A) gene polymorphisms and type 2 diabetes mellitus: a meta-analysis. Diabetes/metabolism research and reviews. 2011; 27(2):177–84. https://doi.org/10.1002/dmrr.1158
PMID: 21294239. 15. Lucia A, Gomez-Gallego F, Barroso I, Rabadan M, Bandres F, San Juan AF, et al. PPARGC1A geno-
type (Gly482Ser) predicts exceptional endurance capacity in European men. Journal of applied physiol-
ogy. 2005; 99(1):344–8. https://doi.org/10.1152/japplphysiol.00037.2005 PMID: 15705733. 16. Gineviciene V, Pranckeviciene E, Milasius, Kucinskas V. Gene variants related to the power perfor-
mance of the Lithuanian athletes. Cent Eur J Biol. 2011; 6(11):48–57. https://doi.org/10.247B/s11535-
010-0102-5 17. Gineviciene V, Jakaitiene A, Aksenov MO, Aksenova AV, Druzhevskaya AM, Astratenkova IV, et al. Association analysis of ACE, ACTN3 and PPARGC1A gene polymorphisms in two cohorts of European
strength and power athletes. Biology of sport. 2016; 33(3):199–206. https://doi.org/10.5604/20831862. 1201051 PMID: 27601773; PubMed Central PMCID: PMC4993134. 18. Jin H, Hwang IW, Kim KC, Cho HI, Park TH, Shin YA, Lee HS, Hwang JH, Kim AR, Lee KH, Shin YE,
Lee JY, Kim JA, Choi EJ, Kim BK, Sim HS, Kim MS and Kim W. Is there a relationship between PPARD
T294C/PPARGC1A Gly482Ser variations and physical endurance performance in the Korean popula-
tion? Genes Genom. 2015. Epub 22 December 2015. https://doi.org/10.1007/s13258-015-0380-4 19. Maciejewska A, Sawczuk M, Cieszczyk P, Mozhayskaya IA, Ahmetov II. The PPARGC1A gene
Gly482Ser in Polish and Russian athletes. Journal of sports sciences. 2012; 30(1):101–13. https://doi. org/10.1080/02640414.2011.623709 PMID: 22122487. 20. Grealy R, Herruer J, Smith CL, Hiller D, Haseler LJ, Griffiths LR. Evaluation of a 7-Gene Genetic Profile
for Athletic Endurance Phenotype in Ironman Championship Triathletes. PloS one. 2015; 10(12):
e0145171. https://doi.org/10.1371/journal.pone.0145171 PMID: 26716680; PubMed Central PMCID:
PMC4696732. 21. Clark MF, Baudouin SV. PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 References A systematic review of the quality of genetic association studies in human sep-
sis. Intensive Care Med. 2006; 32(11):1706–12. https://doi.org/10.1007/s00134-006-0327-y PMID:
16957907. 22. Berthold FH V.M., Klawon F. How to intelligentlymake sense of real data, in descriptive statistics. In:
Springer, editor. Guide to Intelligent Data Analysis: Springer 2010. p. p. 315. 23. Lau J, Ioannidis JP, Schmid CH. Quantitative synthesis in systematic reviews. Ann Intern Med. 1997;
127(9):820–6. PMID: 9382404. 24. Higgins JP, Thompson SG. Quantifying heterogeneity in a meta-analysis. Stat Med. 2002; 21
(11):1539–58. https://doi.org/10.1002/sim.1186 PMID: 12111919. 25. Mantel N, Haenszel W. Statistical aspects of the analysis of data from retrospective studies of disease. J Natl Cancer Inst. 1959; 22(4):719–48. PMID: 13655060. 26. DerSimonian R, Laird N. Meta-analysis in clinical trials. Control Clin Trials. 1986; 7(3):177–88. PMID:
3802833. 27. Galbraith RF. A note on graphical presentation of estimated odds ratios from several clinical trials. Stat
Med. 1988; 7(8):889–94. PMID: 3413368. 28. Dumas-Mallet E, Button KS, Boraud T, Gonon F, Munafo MR. Low statistical power in biomedical sci-
ence: a review of three human research domains. Royal Society open science. 2017; 4(2):160254. https://doi.org/10.1098/rsos.160254 PMID: 28386409; PubMed Central PMCID: PMC5367316. 29. Ioannidis JP, Trikalinos TA. The appropriateness of asymmetry tests for publication bias in meta-analy-
ses: a large survey. CMAJ. 2007; 176(8):1091–6. https://doi.org/10.1503/cmaj.060410 PMID:
17420491; PubMed Central PMCID: PMC1839799. 16 / 18 PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 Meta-analysis PPARGC1A athletic ability sports performance 30. Abramson JH. WINPEPI (PEPI-for-Windows): computer programs for epidemiologists. Epidemiol Per-
spect Innov. 2004; 1(1):6. https://doi.org/10.1186/1742-5573-1-6 PMID: 15606913. 31. Ahmetov II, Williams AG, Popov DV, Lyubaeva EV, Hakimullina AM, Fedotovskaya ON, et al. The com-
bined impact of metabolic gene polymorphisms on elite endurance athlete status and related pheno-
types. Human genetics. 2009; 126(6):751–61. https://doi.org/10.1007/s00439-009-0728-4 PMID:
19653005. 32. Eynon N, Meckel Y, Sagiv M, Yamin C, Amir R, Sagiv M, et al. Do PPARGC1A and PPARalpha poly-
morphisms influence sprint or endurance phenotypes? Scandinavian journal of medicine & science in
sports. 2010; 20(1):e145–50. https://doi.org/10.1111/j.1600-0838.2009.00930.x PMID: 19422653. 33. Gineviciene V, Jakaitiene A, Tubelis L, Kucinskas V. Variation in the ACE, PPARGC1A and PPARA
genes in Lithuanian football players. European journal of sport science. 2012; 14 Suppl 1:S289–95. https://doi.org/10.1080/17461391.2012.691117 PMID: 24444220. 34. He ZH, Hu Y, Li YC, Gong LJ, Cieszczyk P, Maciejewska-Karlowska A, et al. PGC-related gene variants
and elite endurance athletic status in a Chinese cohort: a functional study. References Scandinavian journal of med-
icine & science in sports. 2015; 25(2):184–95. https://doi.org/10.1111/sms.12188 PMID: 25170593. 35. Maruszak A, Adamczyk JG, Siewierski M, Sozanski H, Gajewski A, Zekanowski C. Mitochondrial DNA
variation is associated with elite athletic status in the Polish population. Scandinavian journal of medi-
cine & science in sports. 2012; 24(2):311–8. https://doi.org/10.1111/sms.12012 PMID: 23163620. 36. Muniesa CA, Gonzalez-Freire M, Santiago C, Lao JI, Buxens A, Rubio JC, et al. World-class perfor-
mance in lightweight rowing: is it genetically influenced? A comparison with cyclists, runners and non-
athletes. British journal of sports medicine. 2010; 44(12):898–901. https://doi.org/10.1136/bjsm.2008. 051680 PMID: 18801770. 37. Peplonska B, Adamczyk JG, Siewierski M, Safranow K, Maruszak A, Sozanski H, et al. Genetic variants
associated with physical and mental characteristics of the elite athletes in the Polish population. Scandi-
navian journal of medicine & science in sports. 2017; 27(8):788–800. https://doi.org/10.1111/sms. 12687 PMID: 27140937. 38. Yvert T, Miyamoto-Mikami E, Murakami H, Miyachi M, Kawahara T, Fuku N. Lack of replication of asso-
ciations between multiple genetic polymorphisms and endurance athlete status in Japanese population. Physiological reports. 2016; 4(20). https://doi.org/10.14814/phy2.13003 PMID: 27798356; PubMed
Central PMCID: PMC5099965. 39. Hong EP, Park JW. Sample size and statistical power calculation in genetic association studies. Geno-
mics & informatics. 2012; 10(2):117–22. https://doi.org/10.5808/GI.2012.10.2.117 PMID: 23105939;
PubMed Central PMCID: PMC3480678. 40. Faul F, Erdfelder E, Lang AG, Buchner A. G*Power 3: a flexible statistical power analysis program for
the social, behavioral, and biomedical sciences. Behavior research methods. 2007; 39(2):175–91. PMID: 17695343. 41. Christley R. Power and Error: Increased Risk of False Positive Results in Underpowered Studies. The
Open Epidemiology Journal. 2010; 3:16–9. 42. McLaughlin MJ, Sainani KL. Bonferroni, Holm, and Hochberg corrections: fun names, serious changes
to p values. PM & R: the journal of injury, function, and rehabilitation. 2014; 6(6):544–6. https://doi.org/
10.1016/j.pmrj.2014.04.006 PMID: 24769263. 43. Eynon N, Ruiz JR, Oliveira J, Duarte JA, Birk R, Lucia A. Genes and elite athletes: a roadmap for future
research. The Journal of physiology. 2011; 589(Pt 13):3063–70. https://doi.org/10.1113/jphysiol.2011. 207035 PMID: 21540342; PubMed Central PMCID: PMC3145924. 44. Ahmetov II, Mozhayskaya IA, Flavell DM, Astratenkova IV, Komkova AI, Lyubaeva EV, et al. PPARal-
pha gene variation and physical performance in Russian athletes. European journal of applied physiol-
ogy. 2006; 97(1):103–8. https://doi.org/10.1007/s00421-006-0154-4 PMID: 16506057. 45. Liang H, Ward WF. PGC-1alpha: a key regulator of energy metabolism. Advances in physiology educa-
tion. 2006; 30(4):145–51. https://doi.org/10.1152/advan.00052.2006 PMID: 17108241. 46. PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 References Mathai AS, Bonen A, Benton CR, Robinson DL, Graham TE. Rapid exercise-induced changes in PGC-
1alpha mRNA and protein in human skeletal muscle. Journal of applied physiology. 2008; 105(4):1098–
105. https://doi.org/10.1152/japplphysiol.00847.2007 PMID: 18653753. 47. Meijer JP, Jaspers RT, Rittweger J, Seynnes OR, Kamandulis S, Brazaitis M, et al. Single muscle fibre
contractile properties differ between body-builders, power athletes and control subjects. Experimental
physiology. 2015; 100(11):1331–41. https://doi.org/10.1113/EP085267 PMID: 26388513. 48. Olesen J, Kiilerich K, Pilegaard H. PGC-1alpha-mediated adaptations in skeletal muscle. Pflugers
Archiv: European journal of physiology. 2010; 460(1):153–62. https://doi.org/10.1007/s00424-010-
0834-0 PMID: 20401754. 17 / 18 PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 Meta-analysis PPARGC1A athletic ability sports performance 49. Suzuki K, Nakaji S, Yamada M, Totsuka M, Sato K, Sugawara K. Systemic inflammatory response to
exhaustive exercise. Cytokine kinetics. Exercise immunology review. 2002; 8:6–48. PMID: 12690937. 50. Nikolaidis MG, Jamurtas AZ, Paschalis V, Fatouros IG, Koutedakis Y, Kouretas D. The effect of mus-
cle-damaging exercise on blood and skeletal muscle oxidative stress: magnitude and time-course con-
siderations. Sports medicine. 2008; 38(7):579–606. https://doi.org/10.2165/00007256-200838070-
00005 PMID: 18557660. 51. Powers SK, Jackson MJ. Exercise-induced oxidative stress: cellular mechanisms and impact on muscle
force production. Physiological reviews. 2008; 88(4):1243–76. https://doi.org/10.1152/physrev.00031. 2007 PMID: 18923182; PubMed Central PMCID: PMC2909187. 52. Melloul D, Stoffel M. Regulation of transcriptional coactivator PGC-1alpha. Science of aging knowledge
environment: SAGE KE. 2004; 2004(9):pe9. https://doi.org/10.1126/sageke.2004.9.pe9 PMID:
14999129. 53. Lin J, Wu H, Tarr PT, Zhang CY, Wu Z, Boss O, et al. Transcriptional co-activator PGC-1 alpha drives
the formation of slow-twitch muscle fibres. Nature. 2002; 418(6899):797–801. https://doi.org/10.1038/
nature00904 PMID: 12181572. 54. Norrbom J, Sundberg CJ, Ameln H, Kraus WE, Jansson E, Gustafsson T. PGC-1alpha mRNA expres-
sion is influenced by metabolic perturbation in exercising human skeletal muscle. Journal of applied
physiology. 2004; 96(1):189–94. https://doi.org/10.1152/japplphysiol.00765.2003 PMID: 12972445. 55. Pilegaard H, Saltin B, Neufer PD. Exercise induces transient transcriptional activation of the PGC-
1alpha gene in human skeletal muscle. The Journal of physiology. 2003; 546(Pt 3):851–8. https://doi. org/10.1113/jphysiol.2002.034850 PMID: 12563009; PubMed Central PMCID: PMC2342594. 56. Russell AP, Feilchenfeldt J, Schreiber S, Praz M, Crettenand A, Gobelet C, et al. Endurance training in
humans leads to fiber type-specific increases in levels of peroxisome proliferator-activated receptor-
gamma coactivator-1 and peroxisome proliferator-activated receptor-alpha in skeletal muscle. Diabe-
tes. 2003; 52(12):2874–81. PMID: 14633846. 57. Sczelecki S, Besse-Patin A, Abboud A, Kleiner S, Laznik-Bogoslavski D, Wrann CD, et al. PLOS ONE | https://doi.org/10.1371/journal.pone.0200967
January 9, 2019 References Loss of Pgc-
1alpha expression in aging mouse muscle potentiates glucose intolerance and systemic inflammation. American journal of physiology Endocrinology and metabolism. 2014; 306(2):E157–67. https://doi.org/
10.1152/ajpendo.00578.2013 PMID: 24280126; PubMed Central PMCID: PMC4073996. 58. Esterbauer H, Oberkofler H, Krempler F, Patsch W. Human peroxisome proliferator activated receptor
gamma coactivator 1 (PPARGC1) gene: cDNA sequence, genomic organization, chromosomal locali-
zation, and tissue expression. Genomics. 1999; 62(1):98–102. https://doi.org/10.1006/geno.1999.5977
PMID: 10585775. 59. Ahmetov II, Rogozkin VA. Genes, athlete status and training—An overview. Medicine and sport sci-
ence. 2009; 54:43–71. https://doi.org/10.1159/000235696 PMID: 19696507. 60. Beunen G, Thomis M. Gene driven power athletes? Genetic variation in muscular strength and power. British journal of sports medicine. 2006; 40(10):822–3. https://doi.org/10.1136/bjsm.2006.029116
PMID: 17021009; PubMed Central PMCID: PMC2465051. 61. Thakkinstian A, McElduff P, D’Este C, Duffy D, Attia J. A method for meta-analysis of molecular associ-
ation studies. Stat Med. 2005; 24(9):1291–306. https://doi.org/10.1002/sim.2010 PMID: 15568190. 18 / 18
|
https://openalex.org/W2136060196
|
https://actavet.vfu.cz/media/pdf/avb_2013082040375.pdf
|
English
| null |
Determination of minimum effective doses of luteinizing hormone and human chorionic gonadotropin for intrafollicular treatment to induce ovulation in dairy heifers
|
Acta veterinaria Brno
| 2,013
|
cc-by
| 3,722
|
Abstract The aim of this study was to determine the minimum effective intrafollicular doses of luteinizing
hormone and human chorionic gonadotropin in order to induce ovulation in cycling dairy heifers
that have not yet been adequately established. Application of 10, 5, 1, 0.5, 0.1, 0.01 and 0.001
µg luteinizing hormone as well as 10, 1, 0.1, 0.01 and 0.001 international units (IU) of human
chorionic gonadotropin in dominant follicles was performed on day 7 of the oestrous cycle. Control
animals were given luteinizing hormone (12.5 mg and 25 mg) or human chorionic gonadotropin
(2000 IU) intravenously. Accessory corpus luteum on day 14 of the oestrous cycle was considered
as an evidence of ovulation. Ovulation was observed in 2 out of 3 heifers in each treatment group
(n = 3) after administration of 10–0.1 µg luteinizing hormone (except for 0.5 µg – ovulation in 3 of 3
heifers), in all heifers after administration of 10–0.01 IU human chorionic gonadotropin as well as in
all control heifers. Administration of 0.01 µg and 0.001 µg luteinizing hormone as well as of 0.001 IU
human chorionic gonadotropin did not result in ovulation. Higher progesterone concentration on day
14 vs. day 7 of the oestrous cycle was found after all treatments. Nevertheless, the differences were
significant (P < 0.05) only after intrafollicular treatments with 5, 1 and 0.001 µg luteinizing hormone
as well as 10, 1 and 0.01 IU human chorionic gonadotropin. In conclusion, minimum efficient doses for
intrafollicular treatment of the dominant follicles in cycling heifers capable of inducing ovulation were
0.1 µg of luteinizing hormone and 0.01 IU of human chorionic gonadotropin. This is the first study
describing the intrafollicular luteinizing hormone administration in cycling dairy heifers. Intraovarian injection, ovum pick-up, ultrasound-guided transvaginal ovarian puncture, accessory
corpus luteum Ultrasound-guided transvaginal intrafollicular injection of human chorionic gonadotropin
(hCG) in cattle was first reported as a useful research tool by Kot et al. in 1995. Since
then, intrafollicular injection of different substances such as phosphate-buffered saline or
insulin-like growth factor-I (Bergfelt et al. 1998; Ginther et al. 2004; Shahiduzzaman
et al. 2010) as well as intrafollicular insemination (Lopez-Gatius and Hunter 2011)
have been described. In our former study, intrafollicular treatment (IFT) with luteinizing
hormone (LH) injected in the dominant follicle in heifers previously treated by deslorelin
implants was reported. Ovulation was proven after the intrafollicular treatment using
various doses of LH (Mala et al. 2013). Received October 22, 2012
Accepted August 28, 2013 Received October 22, 2012
Accepted August 28, 2013 Address for correspondence:
Svatopluk Čech
Cattle and Swine Clinic, Faculty of Veterinary Medicine
University of Veterinary and Pharmaceutical Sciences Brno
Palackého 1/3, 612 42 Brno, Czech Republic Abstract However, minimum effective doses of LH for IFT
in comparison with IFT using hCG have not yet been adequately established.i p
g
y
q
y
The aim of this study was to determine the minimum efficient doses of LH and hCG for
intrafollicular treatment to induce ovulation in cycling dairy heifers. ACTA VET. BRNO 2013, 82: 375–379; doi:10.2754/avb201382040375 ACTA VET. BRNO 2013, 82: 375–379; doi:10.2754/avb201382040375 Materials and Methods Intrafollicular treatment equipment Jana Malá1, Jean-Francois Beckers2, Noelita Melo de Sousa2, Eva Indrová1, Miloslava Lopatářová1,
Radovan Doležel1, Helena Ševelová1, Svatopluk Čech1 Jana Malá1, Jean-Francois Beckers2, Noelita Melo de Sousa2, Eva Indrová1, Miloslava Lopa
Radovan Doležel1, Helena Ševelová1, Svatopluk Čech1 1University of Veterinary and Pharmaceutical Sciences, Faculty of Veterinary Medicine,
Ruminant and Swine Clinic, Brno, Czech Republic
2University of Liege, Faculty of Veterinary Medicine, Laboratory of Physiology of Animal Reproduction, Liege, Belgium 1University of Veterinary and Pharmaceutical Sciences, Faculty of Veterinary Medicine,
Ruminant and Swine Clinic, Brno, Czech Republic
Liege, Faculty of Veterinary Medicine, Laboratory of Physiology of Animal Reproduction, Liege, Belgium 1University of Veterinary and Pharmaceutical Sciences, Faculty of Veterinary Medicine,
Ruminant and Swine Clinic, Brno, Czech Republic
2University of Liege, Faculty of Veterinary Medicine, Laboratory of Physiology of Animal Reproduction, Liege, Belgium 1University of Veterinary and Pharmaceutical Sciences, Faculty of Veterinary Medicine,
Ruminant and Swine Clinic, Brno, Czech Republic
of Liege, Faculty of Veterinary Medicine, Laboratory of Physiology of Animal Reproduction, Liege, Belgium ,
,
p
f Liege, Faculty of Veterinary Medicine, Laboratory of Physiology of Animal Reproduction, Liege, Belgium Statistical analysis Progesterone concentrations are presented as a mean value plus standard deviation. Data of P4 values in
cases without accessory CL were not excluded. Comparison of P4 values with the animals bearing the accessory
CL was not performed due to a limited number of animals. Differences between groups were analyzed using
Student’s paired t-test, Fisher’s exact test and Tukey-Kramer test (pairwise comparisons for one-way layout
design). Statistical difference was presented as P < 0.05. Blood sampling and progesterone assay Blood sampling and progesterone assay Peripheral blood was obtained from vena caudalis mediana right before the treatment (D7 of the oestrous
cycle) and 7 days later (D14). Plasma was immediately separated by centrifugation (5 000 g) and stored frozen at
-20 °C. Progesterone (P4) concentrations were determined by direct radioimmunoassay (RIA) method (without
extraction), as described in detail by Lopez-Gatius et al. (2007). The minimum detection limit of the P4-RIA
technique used was 0.15 ng/ml. The intra-assay and inter-assay coefficients of variation of P4 radioimmunoassay
were 13.8% (3.1 ± 0.4 ng/ml) and 19% (2.7 ± 0.5 ng/ml) respectively. Intrafollicular treatment equipment Intrafollicular treatment was performed using a newly developed double channel instrument enabling aspiration of
the follicular fluid and subsequent IFT with exactly the same amount of solution, while the dead volume is only the
volume of the needle. The instrument and intrafollicular treatment were previously described by Cech et al. (2013). Phone: +420 607 214 021
E-mail: cechs@vfu.cz
http://actavet.vfu.cz/ 376 Animals, treatment and ultrasound schedule Animals, treatment and ultrasound schedule Animals, treatment and ultrasound schedule Holstein heifers (n = 45) at the age of 13 months and 350 kg body weight were used for the experiment. Heifers were kept at commercial dairy farms under usual conditions; the experiment was approved by the ethics
committee (No 82/2012). Heifers bearing the corpus luteum (CL) were synchronized by cloprostenol (500 μg
i.m. pro toto, Oestrophan®, Bioveta a.s., Czech Republic). After synchronization, the day when females exhibited
standing oestrus was considered as day 0 (D0). On day 7 of the cycle (D7) when the first follicular wave appeared
clearly, the dominant follicle was submitted to intrafollicular treatment. Mostly one follicle was present; when
two follicles were found, the larger one was treated. The minimum size of treated follicle was 10 mm. Seven
different LH doses (10, 5, 1, 0.5, 0.1, 0.01 and 0.001 µg) (LH, prof. Beckers, Universite de Liege, Belgium) and
five different hCG doses (10, 1, 0.1, 0.01 and 0.001 IU) (Pregnyl®, Organon, Netherlands) were administered in
treatment groups, each consisting of three heifers. Follicular collapse was observed immediately after IFT in two
heifers that were excluded from the trial. Intravenous treatment was performed in control heifers using 12.5 mg of LH (n = 2), 25 mg of LH (n = 2;
Lutropin®, Bioniche, Canada) or 2000 IU hCG (n = 3). Ovaries were scanned on D7 and D14 of oestrous cycle using a real-time B-mode ultrasound machine (SSD-
500, Aloka, Japan) equipped with a linear ultrasound transducer (7.5 MHz Aloka UST 5561, Japan). Development
of an accessory CL was considered as a positive response after IFT. Results In addition, the
progesterone concentration
on D7 in heifers treated with
1 µg LH was significantly
(P < 0.05) different from
heifers treated with 0.1 µg and
0.01 µg LH and progesterone
concentration on D7 in
heifers intravenously treated
with 2000 IU hCG was
significantly (P < 0.05)
different from heifers treated
with intrafollicular injection
of 0.01 µg and 0.001 IU
hCG. Table 2. Positive response (accessory corpus luteum) of ovaries seven
days after intrafollicular and intravenous human chorionic gonadotropin
treatment in dairy heifers. hCG - human chorionic gonadotropin, CL - corpus luteum,
IFT - intrafollicular treatment, i.v. – intravenous treatment
Treatment
hCG dose (IU)
n
Accessory CL
IFT
10
3
3
1
3
3
0.1
3
3
0.01
3
3
0.001
3
0
Control i.v. 2000
3
3 Table 3. Progesterone concentrations (ng/ml) on day 7 and day 14 of the
oestrous cycle (day of intrafollicular treatment and seven days later) using
different doses of luteinizing hormone and after i.v. luteinizing hormone
treatment. Table 3. Progesterone concentrations (ng/ml) on day 7 and day 14 of the
oestrous cycle (day of intrafollicular treatment and seven days later) using
different doses of luteinizing hormone and after i.v. luteinizing hormone
treatment. IFT - intrafollicular treatment, LH - luteinizing hormone, a, bvalues in
D14 column are significantly different (P < 0.05), *values in rows are
significantly different (P < 0.05), i.v. – intravenous treatment
Treatment
D7
D14
IFT 10 µg LH
4.38 ± 1.21
7.39 ± 1.52a,b
IFT 5 µg LH
2.09 ± 0.24
12.82 ± 1.32a,b*
IFT 1 µg LH
4.29 ± 1.05
14.61 ± 2.74a*
IFT 0.5 μg LH
4.39 ± 0.2
7.39 ± 4.72a,b
IFT 0.1 μg LH
2.75 ± 0.53
5.94 ± 1.89b
IFT 0.01 μg LH
2.71 ± 2.03
5.9 ± 1.43b
IFT 0.001 μg LH
2.68 ± 1.09
7.45 ± 1.54a,b*
i.v. 12.5–25 mg LH
3.07 ± 1.17
9.11 ± 0.77a,b Results Ovulation (accessory CL on D7) was observed in 2 out of 3 heifers after IFT at doses of
10, 5, 1 and 0.1 µg LH. Positive ovarian response occurred and in all animals after IFT at
the dose of 0.5 µg LH and after i.v. LH treatment. The doses of 0.01 and 0.001 µg did not
lead to ovulation (Table 1). P
i i
i Positive ovarian response
was observed in all heifers (3
out of 3) after IFT at doses of
10, 1, 0.1 and 0.01 IU hCG
as well as after intravenous
hCG treatment. The dose
of 0.001 IU did not lead to
ovulation (Table 2). Table 1. Positive response (accessory corpus luteum) of ovaries seven
days after intrafollicular and intravenous luteinizing hormone treatment
in dairy heifers. LH - luteinizing hormone, CL - corpus luteum, IFT - intrafollicular
treatment, i.v. – intravenous treatment
Treatment
LH dose (μg)
n
Accessory CL
IFT
10
3
2
5
3
2
1
3
2
0.5
3
3
0.1
3
2
0.01
3
0
0.001
3
0
Control i.v. 12.5
2
2
25
2
2
in dairy heifers. (
)
In terms of all positively
responding
groups,
total
efficiency of IFT using LH
and hCG was 73.3% (11 out
of 15) and 100% (12 out of
12), respectively. However,
the
difference
was
not
significant (P > 0.05). Increased
progesterone
concentrations
on
D14 377 compared to D7 (immediately
before treatment) were found
after all treatments. The
differences were significant
(P < 0.05) in groups of heifers
treated with intrafollicular
application of 5, 1 and 0.001
µg LH (Table 3) and in
groups of heifers treated with
intrafollicular
application
of 10, 1 and 0.01 IU hCG
(Table 4). In addition, the
progesterone concentration
on D7 in heifers treated with
1 µg LH was significantly
(P < 0.05) different from
heifers treated with 0.1 µg and
0.01 µg LH and progesterone
concentration on D7 in
heifers intravenously treated
with 2000 IU hCG was
significantly (P < 0.05)
different from heifers treated
with intrafollicular injection
of 0.01 µg and 0.001 IU
hCG. compared to D7 (immediately
before treatment) were found
after all treatments. The
differences were significant
(P < 0.05) in groups of heifers
treated with intrafollicular
application of 5, 1 and 0.001
µg LH (Table 3) and in
groups of heifers treated with
intrafollicular
application
of 10, 1 and 0.01 IU hCG
(Table 4). Discussion 1995) in which precise calculations of hCG doses for IFT were described. The effective intrafollicular dose was approximately 10 000 × lower than the systemic dose
allowing ovulation (0.01 IU hCG). The range of LH doses was established with regard to
our previous study in which the IFT with doses of 10, 5 and 1 µg LH were able to induce
ovulation, whereas the dose of 0.01 µg LH was not followed by ovulation in dairy heifers
treated by deslorelin (Mala et al. 2013). Based on these data, it was decided to use an
array of decreasing doses of LH (from 10 to 0.001 µg) and hCG (from 10 to 0.001 IU). The results were not compared with the effect of IFT with saline, as its inability to induce
ovulation in dairy cows was proved sufficiently (Cech et al. 2013). Intrafollicular treatment using LH and hCG in the abovementioned doses proved to be
highly efficient in inducing ovulation in cycling heifers. High efficiency of 66.7% was
found after administration of 10, 1 and 0.1 µg of LH, 100% efficiency was observed after
administration of 5 µg LH as well as 100% efficiency was noted after administration of
10, 1, 0.1 and 0.01 IU hCG. Only doses of 0.01 and 0.001 µg LH and 0.001 IU hCG failed
to induce ovulation. The differences among separate groups were not significant due to
insufficient number of animals. Because of the limited number of animals in the groups we
compared a total efficiency of IFT with different doses of LH and hCG. After estimation of
the minimum effective doses we involved only effective doses into the comparison.i y
p
Overall, total efficiency in positively responding groups was numerically lower after LH
IFT (73.3%) compared to hCG IFT (100%). These differences are remarkable but difficult
to explain. Unsuccessful treatments could not be caused by technical problems with IFT, as
all treated heifers enrolled in the study showed a clear turbulence in treated follicles and, at
the same time, intrafollicular injections were performed gently and without any difficulties. All treated follicles were prepared as per the same synchronization protocol and they
were in the same maturation stage. In addition, higher efficiency of LH was found in the
previous study, where all heifers treated by deslorelin showed ovulation after intrafollicular
treatment at the dose of 5 as well as 10 µg LH (Mala et al. 2013). Discussion The effects of different
doses
of
intrafollicular
administration of hCG as
well as the technical aspects
of IFT were described in
the first report concerning
ultrasound-guided
intra-
follicular
injection
(Kot
et al. 1995). The effect of
intrafollicular injection of LH
in heifers previously treated
with deslorelin was recently
evaluated by Mala et al. (2013). However, minimum
effective intrafollicular doses
of LH in comparison with
intrafollicular administration
of hCG in cycling heifers
were not established. In this
experiment, the range of
hCG doses was determined
with regard to an earlier IFT - intrafollicular treatment, LH - luteinizing hormone, a, bvalues in
D14 column are significantly different (P < 0.05), *values in rows are
significantly different (P < 0.05), i.v. – intravenous treatment Table 4. Progesterone concentrations (ng/ml) on day 7 and day 14 of
oestrous cycle (day of intrafollicular treatment and seven days later) using
different doses of human chorionic gonadotropin and after i.v. human
chorionic gonadotropin treatment. Table 4. Progesterone concentrations (ng/ml) on day 7 and day 14 of
oestrous cycle (day of intrafollicular treatment and seven days later) using
different doses of human chorionic gonadotropin and after i.v. human
chorionic gonadotropin treatment. Treatment
D7
D14
IFT 10 IU hCG
4.07 ± 1.57
13.82 ± 2.84a,b*
IFT 1 IU hCG
4.37 ± 1.88
12.19 ± 0.98a,b*
IFT 0.1 IU hCG
2.37 ± 0.64
11.33 ± 3.85a,b
IFT 0.01 IU hCG
2.67 ± 0.25
7.33 ± 0.52b*
IFT 0.001IU hCG
2.85 ± 1.6
6.73 ± 1.02b
i.v. 2000 IU hCG
4.9 ± 1.55
18.93 ± 4.78a IFT - intrafollicular treatment, hCG – human chorionic gonadotropin, a,b
values in D14 column are significantly different (P < 0.05), * values in
rows are significantly different (P < 0.05), i.v. – intravenous treatment
Treatment
D7
D14
IFT 10 IU hCG
4.07 ± 1.57
13.82 ± 2.84a,b*
IFT 1 IU hCG
4.37 ± 1.88
12.19 ± 0.98a,b*
IFT 0.1 IU hCG
2.37 ± 0.64
11.33 ± 3.85a,b
IFT 0.01 IU hCG
2.67 ± 0.25
7.33 ± 0.52b*
IFT 0.001IU hCG
2.85 ± 1.6
6.73 ± 1.02b
i.v. 2000 IU hCG
4.9 ± 1.55
18.93 ± 4.78a IFT - intrafollicular treatment, hCG – human chorionic gonadotropin, a,b
values in D14 column are significantly different (P < 0.05), * values in
rows are significantly different (P < 0.05), i.v. – intravenous treatment 378 study (Kot et al. Discussion The efficiency might
be affected also by different half-life of LH and hCG. And that is why lower efficiency
of LH in comparison with hCG in this study should be confirmed with a larger number
of treated animals. Efficiency of IFT treatment with hCG in the present study was similar
to that previously described in the study by Kot et al. (1995). The results of the present
study confirmed the same minimum effective dose of 0.01 UI hCG for intrafollicular
administration. Kot et al. (1995) described ovarian cysts after IFT which was not followed by ovulation. However, in our study we did not observe the occurrence of ovarian cysts in any heifer. Post-treatment ultrasonography of CLs was typical, except for two heifers after IFT
(1 µg LH and 0.1 IU hCG) and 1 heifer after intravenous administration of 12.5 mg
LH. Accessory corpora lutea of these heifers contained a large cavity and a very thin
(1–2 mm) layer of luteal tissue. Although these cases were classified as a positive response,
the structures themselves were not an unambiguous evidence of ovulation due to the
possibility of luteal cysts. Higher progesterone concentrations were found on D14 of the oestrous cycle compared
to D7 after all treatments. The differences were significant in groups of heifers treated
with 5, 1 and 0.001 LH and in groups of females treated with 10, 1 and 0.01 IU hCG. Evaluation of progesterone concentrations was difficult due to the limited number of
animals in the experimental groups and to large variability of progesterone values. The
increase of progesterone could be caused by secretion of accessory corpora lutea and/or
by increased secretory activity of original CL after the treatment. Significant increase of
progesterone in heifers without ovulatory response after IFT with 0.001 μg LH proved
the importance of secretory activity of original CL in progesterone concentration after the
treatment. Increased progesterone concentrations due to accessory corpora lutea and/or 379 higher secretory activity of original CL after GnRH administered to heifers or cows within
several days after ovulation were described by Garcia-Ispierto and Lopez-Gatius
(2012) and Kasimanickam et al. (2011). (
)
(
)
Progesterone concentration on D14 of the oestrous cycle (seven days after IFT) was
significantly higher in heifers treated with 1 µg LH compared to heifers treated with
0.1 µg and 0.01 µg LH. Acknowledgements The study was supported by IGA VFU Brno 6/2011/FVL. The study was supported by IGA VFU Brno 6/2011/FVL. Discussion Progesterone concentration on the seventh day after intravenous
administration of 2000 UI hCG (D14 of oestrous cycle) was significantly higher compared
to IFT with 0.01 µg and 0.001 IU hCG. The highest absolute value of progesterone in the
control group seven days after intravenous administration of hCG could be caused by the
direct stimulatory effect of hCG on original CL. Progesterone concentrations after IFT
showed a clear downward tendency with decreasing doses of hCG, although the heifers
showed a positive ovarian response. It was assumed that higher intrafollicular doses of
hormones could be followed by development of original and/or accessory corpus luteum
and higher progesterone secretion. For this reason, progesterone secretion after IFT should
be taken into account as the second indicator for the recommended dose of LH or hCG. Considering progesterone concentrations, the recommended minimum doses appears to be
higher (1 µg of LH and 10 IU of hCG) than the dose required for induction of ovulation.i g
( µg
)
q
In conclusion, the minimum efficient doses for intrafollicular treatment to induce
ovulation of dominant follicles in cycling heifers were 0.1 µg of LH and 0.01 IU of hCG. Nevertheless, the doses of 1 µg LH and 10 IU hCG are recommended to ensure sufficient
progesterone secretion after the treatment. Further studies on efficacy of IFT with LH and
hCG as well as secretory activity of accessory corpora lutea after IFT using different doses
of LH and hCG should be conducted with a larger number of experimental heifers. References Bergfelt DR, Brogliatti GM, Adams GP 1998: Gamete recovery and follicular transfer (GRAFT) using transvaginal
ultrasonography in cattle. Theriogenology 50: 15-25 Bergfelt DR, Brogliatti GM, Adams GP 1998: Gamete recovery and follicular transfer (GRAFT) using transvaginal
ultrasonography in cattle. Theriogenology 50: 15-25 g p y
g
gy
Cech S, Mala J, Indrova E, Lopatarova M, Dolezel R, Dluhosova H, Zilka L 2013: The introduction of a double-
channel system for the intrafollicular treatment of cattle. Vet Med-Czech 58: 10-15 Garcia-Ispierto I, Lopez-Gatius F 2012: Effects of GnRH or progesterone treatment on day 5 post-AI on plasma
progesterone, luteal blood flow and leucocyte counts during the luteal phase in dairy cows. Reprod Domest
Anim 47: 224-229 Ginther OJ, Bergfelt DR, Beg MA, Meira C, Kot K 2004: In vivo effects of an intrafollicular injection of insulin-
like growth factor 1 on the mechanism of follicular deviation in heifers and mares. Biol Reprod 70: 99-105 Kasimanickam RK, Mink MR, Nebel RL 2011: Effect of GnRH administration at day 5 or day 7 after AI on
progesterone concentration, corpus luteum volume and pregnancy in lactating dairy cows. Clin Theriogenology
3: 455-459 Kot K, Gibbons JR, Ginther OJ 1995: A technique for intrafollicular injection in cattle: Effects of hCG. Theriogenology 44: 41-50 Lopez-Gatius F, Garbayo JM, Santolaria P, Yániz J, Ayad A, de Sousa NM, Beckers JF 2007: Milk production
correlates negatively with plasma levels of pregnancy-associated glycoprotein (PAG) during the early fetal
period in high producing dairy cows with live fetuses. Domest Anim Endocrin 32: 29-42 p
g p
g
y
López-Gatius F, Hunter RHF 2011: Intrafollicular insemination for the treatment of infertility in the dairy cow. Theriogenology 75: 1695-1698 g
gy
Beckers JF, Sousa NM, Indrova E, Lopatarova M, Dolezel R, Cech S 2013: Intrafollicular LH
stration in dairy heifers treated by GnRH agonist. Vet Med-Czech 58: 81-86 y
y
g
Shahiduzzaman AKM, Beg MA, Palhao MP, Siddiqui MAR, Shamsuddin M, Ginther OJ 2010: Stimulation of
the largest subordinate follicle by intrafollicular treatment with insulin-like growth factor 1 is associated with
inhibition of the dominant follicle in heifers. Theriogenology 74: 194-201
|
https://openalex.org/W2108676843
|
https://dash.harvard.edu/bitstream/1/11855922/1/3731267.pdf
|
English
| null |
AAV-Mediated Administration of Myostatin Pro-Peptide Mutant in Adult Ldlr Null Mice Reduces Diet-Induced Hepatosteatosis and Arteriosclerosis
|
PloS one
| 2,013
|
cc-by
| 9,751
|
Terms of Use This article was downloaded from Harvard University’s DASH repository, and is made available
under the terms and conditions applicable to Other Posted Material, as set forth at http://
nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#LAA Permanent link http://nrs.harvard.edu/urn-3:HUL.InstRepos:11855922 Published Version
doi:10.1371/journal.pone.0071017 Published Version
doi:10.1371/journal.pone.0071017 Citation Guo, Wen, Siu Wong, and Shalender Bhasin. 2013. “AAV-Mediated Administration of Myostatin
Pro-Peptide Mutant in Adult Ldlr Null Mice Reduces Diet-Induced Hepatosteatosis and
Arteriosclerosis.” PLoS ONE 8 (8): e71017. doi:10.1371/journal.pone.0071017. http://
dx.doi.org/10.1371/journal.pone.0071017. Materials and Methods We show here that administration of a protease-resistant
myostatin pro-peptide D76A mutant to adult mice, using the
AAV technology, induced a substantial reduction in liver fat
infiltration and aortic atheromatous lesions with only a mild
impact on body fat and lean mass. The strong association between Share Your Story The Harvard community has made this article openly available. Please share how this access benefits you. Submit a story . Accessibility Abstract This is an open-access article distributed under the terms of the Creative Commons A
use, distribution, and reproduction in any medium, provided the original author and source are credited. Copyright: 2013 Guo et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work was supported by National Institutes of Health grant RO1 DK059261 (WG) and DK078512 (SB). The funders had no role in study design, data
collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. Funding: This work was supported by National Institutes of Health grant RO1 DK059261 (WG) and DK078512 (SB). The funders had no role in study design, data
collection and analysis, decision to publish, or preparation of the manuscript. Funding: This work was supported by National Institutes of Health grant RO1 DK059261 (WG) and DK078512 (SB). The funders had n
collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. * E-mail: wguo2@partners.org Introduction this
metabolically
favorable
phenotype
and
robust
ectopic
expression of myostatin pro-peptide in the liver of the D76A-
treated mice, with minor changes in muscle and fat mass, led us to
consider the hypothesis that myostatin may have direct effects on
hepatic lipid metabolism. We show here for the first time that
myostatin increases de novo hepatic lipogenesis in cultured liver
cells. In addition, we show that myostatin and its receptor are both
abundantly
expressed
in
mouse
aorta. Exposure
of
aortic
endothelial cells to myostatin resulted in activation of TGFb
signaling and reduced phosphorylation of endothelial NO synthase
(eNOS) in association with increased expression of pro-athero-
genic adhesion molecules ICAM-1 and VCAM-1. Our results
indicate that both liver and endothelium are direct targets of
myostatin which may be involved in diet-induced metabolic
disorders. Metabolic disorders, such as diabetes mellitus, nonalcoholic
fatty liver disease, and arteriosclerosis are leading causes of
morbidity and mortality in modern world. With the aging of
human populations, the prevalence of these inter-linked metabolic
disorders is increasing globally. We show here that myostatin, a
muscle-secreted growth and differentiation factor, may be a
potential therapeutic target for the prevention and treatment of
these metabolic disorders. Genetic disruption of myostatin gene
causes marked hypermuscularity and hypoadiposity [1–4]. Genetic
inactivation of myostatin in Ldlr null mice also alleviates diet-
induced hepatosteatosis and arteriosclerosis [5]. However, it is not
known whether similar metabolic improvement can be achieved
by myostatin antagonists in adults, an issue that is crucially
important for the relevant clinical applications. Wen Guo1,2*, Siu Wong1, Shalender Bhasin2 1 Department of Medicine, Boston University School of Medicine, Boston, Massachusetts, United States of America, 2 Research Program in Men’s Health: Aging and
Metabolism, Boston Claude D. Pepper Older Americans Independence Center for Function Promoting Anabolic Therapies, Brigham and Women’s Hospital, Harvard
Medical School, Boston, Massachusetts, United States of America Abstract Genetic disruption of myostatin or its related signaling is known to cause strong protection against diet-induced metabolic
disorders. The translational value of these prior findings, however, is dependent on whether such metabolically favorable
phenotype can be reproduced when myostatin blockade begins at an adult age. Here, we reported that AAV-mediated
delivery of a myostatin pro-peptide D76A mutant in adult mice attenuates the development of hepatic steatosis and
arteriosclerosis, two common diet-induced metabolic diseases. A single dose of AAV-D76A in adult Ldlr null mice resulted in
sustained expression of myostatin pro-peptide in the liver. Compared to vehicle-treated mice, D76A-treated mice gained
similar amount of lean and fat mass when fed a high fat diet. However, D76A-treated mice displayed significantly reduced
aortic lesions and liver fat, in association with a reduction in hepatic expression of lipogenic genes and improvement in liver
insulin sensitivity. This suggests that muscle and fat may not be the primary targets of treatment under our experimental
condition. In support to this argument, we show that myostatin directly up-regulated lipogenic genes and increased fat
accumulation in cultured liver cells. We also show that both myostatin and its receptor were abundantly expressed in
mouse aorta. Cultured aortic endothelial cells responded to myostatin with a reduction in eNOS phosphorylation and an
increase in ICAM-1 and VCAM-1 expression. Conclusions: AAV-mediated expression of myostatin pro-peptide D76A mutant
in adult Ldlr null mice sustained metabolic protection without remarkable impacts on body lean and fat mass. Further
investigations are needed to determine whether direct impact of myostatin on liver and aortic endothelium may contribute
to the related metabolic phenotypes. Citation: Guo W, Wong S, Bhasin S (2013) AAV-Mediated Administration of Myostatin Pro-Peptide Mutant in Adult Ldlr Null Mice Reduces Diet-Induced
Hepatosteatosis and Arteriosclerosis. PLoS ONE 8(8): e71017. doi:10.1371/journal.pone.0071017
Editor: Manlio Vinciguerra, University College London, United Kingdom
Received April 2, 2013; Accepted July 1, 2013; Published August 1, 2013
Copyright: 2013 Guo et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted
use distribution and reproduction in any medium provided the original author and source are credited tation: Guo W, Wong S, Bhasin S (2013) AAV-Mediated Administration of Myostatin Pro-Peptide Mutant in Adult Ldlr Null Mice
patosteatosis and Arteriosclerosis. PLoS ONE 8(8): e71017. doi:10.1371/journal.pone.0071017 ved April 2, 2013; Accepted July 1, 2013; Published August 1, 2 Copyright: 2013 Guo et al. August 2013 | Volume 8 | Issue 8 | e71017 Aortic Lesion Identification After euthanasia, the aorta was stained en face with Sudan IV for
lipid-rich lesions as described before [5,12]. Degree of atheroscle-
rosis in the exposed aorta was presented as the percentage of
lesion-covered area in the tissue surface using NIH Image J
program. Sections of the aortic root (5 mm) near the sinus were
stained with hematoxylin and eosin (H&E) and immuno-stained
against MAC-3 and VCAM-1, as described [5,12]. Results of
immuno-staining was scored blindly following a modified score
system
as
described
before
(http://www.ihcworld.com/
ihc_scoring.htm). Masson’s Trichrome staining was performed
using commercial reagents (Market Lab Inc. Caledonia, MI). Animals and Diet The animal protocol was approved by IACUC of Boston
University. Male Ldlr null mice were purchased from Jackson
Laboratory (Bar Harbor, MI). At eight week of age, mice were
analyzed by NMR and divided into two groups of similar body
composition. Animals were injected through tail vein with either
vehicle (saline) or AAV-D76A (461011 vg/ea, diluted in saline), a
dose similar to those used in previous studies [7]. AAV with no
functional gene products does not cause phenotypic changes in
Ldlr2/2 mice [11]. This was confirmed in our preliminary studies
with AAV-eGFP (not shown). Animals were maintained on regular
chow diet for 7 weeks to stabilize post-injection body composition. All mice were then switched to a high fat diet (TD.09547, Harlan
Laboratories, Madison WI) for 12 weeks. This diet contains
48.4%, 21.4%, and 30.1% calories from carbohydrate, protein,
and fat, respectively, as well as 0.05% cholesterol. Materials Myostatin pro-peptide D76A mutant fused with mouse IgG-Fc
was a gift from Dr. Se-Jin Lee (Johns Hopkins University, August 2013 | Volume 8 | Issue 8 | e71017 August 2013 | Volume 8 | Issue 8 | e71017 PLOS ONE | www.plosone.org 1 Myostatin, Hepatosteatosis, and Atherosclerosis reports that AAV9, when injected through the tail vein, is
primarily expressed in the liver [9,13,14]. Baltimore, MD [6]). The recombinant product of this construct
(D76A-Fc) has been shown to moderately increase skeletal muscle
mass in adult mice [6,7]. The construct was used to generate an
adeno-associated virus (AAV9), using a commercial service (Vector
Biolabs, Philadelphia, PA), for sustained ectopic expression of the
myostatin pro-peptide D76A mutant [7–9]. Recombinant myos-
tatin mature peptide of mammalian origin was provided by
Amgen Inc (Thousand Oaks, CA). Luciferase reporter for TGFb/
myostatin signaling (3TP-Lux) was obtained from Addgene
(#11767, [10]). Renilla luciferase vector (#E6891) and dual
luciferase reporter assay kit (#E1910) were from Promega
(Madison, WI). reports that AAV9, when injected through the tail vein, is
primarily expressed in the liver [9,13,14]. Immunohistochemistry Mouse aorta root was fixed in formalin and sections were
prepared as described before [12]. Immunohistochemistry was
performed using a Zymed HistoMouse-SP Kit (AEC, Broad
Spectrum, Invitrogen #959544), following the manufacturer’s
instructions. Antibody for MAC3 was purchased from BD
Biosciences (#550292; San Jose, CA). Antibody for VCAM-1
was from Santa Cruz (#SC-1504, Santa Cruz, CA). Insulin Tolerance and Pyruvate Tolerance These tests were performed during week 11 after introduction of
the high fat diet. Briefly, insulin (Humalog, USP) was diluted in
sterile saline containing 0.1% BSA and injected (0.6 U/kg, 0.1 ml,
i.p.) after 4 h fasting. Sodium pyruvate (Gibco #11360-070) was
diluted in saline and injected (1.5 g/kg, 0.1 ml, i.p.) after overnight
fasting. Blood glucose was measured at baseline and every 15 min
after the injection for 2 hours. Assays for Fatty Acid Oxidation and De Novo Fatty Acid
Synthesis y
HepG2 cells were pre-labeled with trace amount of [9,10-3H]
oleate overnight (2 mCi per well in a 6 well plate), washed to
remove unincorporated tracers and incubated with myostatin
(100 ng/ml) for 24 h. Cells were washed again and incubated with
serum-free
medium
containing
the
same
concentrations
of
myostatin for another 8 hours. Acid-soluble metabolites released
from the cells were counted as the index for fatty acid oxidation
[17]. Similarly, HepG2 cells were incubated with myostatin for 24
hours and [1,2-14C] acetic acid was added for the last 3 hours of
the incubation. Lipid extracts were separated by TLC (Hexane :
ether : acetic acid at a ratio of 70:30:1) and visualized under iodine
vapor. The triglyceride-incorporated radioactivity was counted as
an index of fatty acid synthesis. Total cellular triglyceride content
was measured after 48 hour incubation and normalized to cellular
protein. Cell Culture Human hepatoma HepG2 cells (a gift from Dr. Mengwei Zang,
Boston University) were grown as described [15]. Cells were
switched to low glucose DMEM (Gibco # 11885084) containing
1% fetal bovine serum (Gibco # 10438-018) overnight and then
treated with myostatin for different incubation periods and
harvested for Western or RT-PCR Analysis. Bovine aortic
endothelial cells (a gift from Dr. Yasuo Ido, Boston University)
were grown as described [16] in low glucose DMEM supplement-
ed with 10% calf serum (Sigma-Aldrich, #12133C) and primary
human aortic endothelial cells (Invitrogen, #C-006-5C) were
grown in a vendor-specified medium (Invitrogen, #C00625PA). Cells were switched to serum-free medium overnight before
treatments. Plasma Lipids Fasting blood samples (no food between 10 pm to 10 am) were
collected 10 weeks after high fat diet. Plasma lipids were measured
as described previously [5,12]. Other Tissue Analysis Selected fat depots, muscle groups, and liver were weighed and
expressed as a percentage of total body weight. Liver lipid staining
and tissue analyses for protein and RNA expression were
performed as described [5,12]. Expression of D76A in liver and
muscle was measured using real-time PCR using primers designed
for myostatin pro-peptide (NM_010834, nt 222–414, which
encompasses
the
D76A
mutation
site). For
the
liver,
the
endogenous myostatin expression was low. Administration of
AAV-D76A induced a large increase in the expression level of pro-
peptide mRNA (Figure S1), a clear demonstration of robust
expression of the ectopic gene. In contrast, we were not able to
detect significant increase in muscle expression of myostatin pro-
peptide (Figure S1, right panel) or mature peptide (primer set for
nt 1015–1145, NM_010834, data not shown). Because full-length
myostatin (a single gene that encodes both pro-peptide and mature
peptide) is strongly expressed in muscle, the lack of difference in
pro-peptide expression between vehicle and D76A-treated mice
indicates that either the ectopic gene was not expressed or its
expression was low enough not to cause a detectable addition to
the endogenous gene. These results are consistent with previous Effects of D76A on Body Composition Consistent with previous reports [7], we observed a modest
increase in lean body mass in mice following the injection of AAV-
D76A while the mice were maintained on a low fat chow (Figure
S2A). Seven weeks after the injection of AAV-D76A, the animals
were switched to a high fat diet to study the effect of D76A on diet-
induced metabolic responses. During the period of high fat diet,
the changes in lean and fat mass were similar between the vehicle
and D76A-treated animals (Figure S2B). Mice were euthanized
after twelve weeks of high fat diet, and individual tissues were
weighed. D76A-treated mice showed a 15% increase in quadriceps
and 5–10% increase in gastrocnemius and levator muscle groups
compared to the control group (Figure S2C). These differences
were relatively small as compared with the 100–150% increase
reported in the myostatin null mice [1–5] and were likely acquired
during the initial low fat feeding period. D76A-treated mice
showed a small decrease in inguinal fat but fat mass in perirenal,
epididymal, and mesenteric depots were not different between the
two groups (Figure S2C). This is in striking contrast to the global
hypoadiposity found in mice with life-long genetic myostatin
blockade [1–5,18], but in line with a recent study on diet-induced
metabolic disorders in mice with adult age myostatin deletion [19]. Since a proatherogenic plasma lipid profile and insulin
resistance are often associated [20], we investigated the effects of
D76A treatment on insulin sensitivity. Fasting glucose (111612 vs. 107613 mg/dl, respectively) and fasting insulin concentrations
(2.9560.7 vs. 2.860.5 ng/ml, respectively) were similar between
the two groups. However, in the non-fasting state (4 h after food
removal in the morning), plasma glucose concentration was lower
in D76A-treated mice than in the controls (Figure 4B, left panel). Since mice were provided free food access except for occasional
fasting (two days out of 3 months), non-fasting results would be
more physiologically relevant, which suggests that D76A-treated
animals have greater steady-state insulin sensitivity than the
control animals. In response to a bolus injection of insulin, the kinetic change of
blood glucose did not differ between the two groups, but the
plasma glucose concentrations decreased to a lower level in the
D76A-treated mice than in the control mice. The area-under-the-
glucose-curve was significantly lower in D76A-treated mice than in
the vehicle-treated mice (AUC glucose 11629 vs. 17880 mg6min/
dL for D76A- and vehicle-treated mice, respectively, p = 0.006). Western Analysis Tissue and cellular homogenates were prepared for Western
analysis as described before [12]. First antibodies were purchased
from Cell Signaling (eNOS, #9586; phosphor-eNOS-Ser1177, August 2013 | Volume 8 | Issue 8 | e71017 2 PLOS ONE | www.plosone.org Myostatin, Hepatosteatosis, and Atherosclerosis #9570; phosphor-Smad3, #9520) and Santa Cruz (VCAM-1,
#SC-1504; beta-tubulin, #SC-9104; ActRIIB, #SC-5665; and
Smad2/3, #SC-6032). adhesion of circulating inflammatory cells, an event facilitated
by the vessel wall production of vascular cell adhesion molecule-1
(VCAM-1), we also stained the aortic root sections for VCAM-1. As shown in Figure 2B, D76A treatment reduced VCAM-1
staining by 30% compared to vehicle treatment. Effects of D76A on Plasma Lipid Profile and Insulin
Sensitivity y
Plasma cholesterol circulates in different forms of lipoproteins,
of which HDL is generally anti-atherogenic and the non-HDL
lipoproteins with variable triglyceride to cholesterol ratios are
proatherogenic. A high ratio between non-HDL and HDL
cholesterol usually correlates with accelerated lesion development,
and vice versus. As shown in Figure 3A, D76A treatment was
associated with lower plasma levels of triglycerides (p = 0.05), total
cholesterol (p = 0.03), and non-HDL cholesterol (p = 0.002), and a
trend towards higher HDL cholesterol (p = 0.06) and lower free
fatty acids (p = 0.08), in comparison to vehicle treatment. The
difference in each measurement, however, is relatively small as
compared to that observed in mice with genetic myostatin
knockout [5], considering that D76A-treatment correlates with a
greater suppression of aortic lesions. Whether D76A specifically
regulates subclass of pro-atherogenic plasma components, such as
oxidized lipoproteins, requires for further investigations. Statistical Analysis All numerical results are presented as means6SEM. Compar-
ison between two treatments were performed using Student’s t test. Comparisons among multiple groups were performed using
ANOVA. Unless specified, all results obtained in tissue culture
experiments are representative of at least three experiments and
results obtained in tissue samples are representative of 5–10
animals of each group. Effects of D76A on Plasma Lipid Profile and Insulin
Sensitivity Effects of D76A on Body Composition These results suggest that D76A-treated mice had better glycemic
control under steady-state condition but their acute response to
insulin administration was not dissimilar from that in vehicle-
treated animals. Because insulin-stimulated glucose uptake occurs
primarily in skeletal muscle, these results suggest that treatment
with D76A likely did not have a major impact on skeletal muscle
insulin sensitivity. Western analysis for selected key elements in the
insulin signaling cascade, including phosphorylated Akt and
p70S6, in the skeletal muscle and the adipose tissue revealed no
significant difference between the two groups (not shown). These
results contrast the findings in mice with genetic myostatin
blockade for which skeletal muscle is hyper-sensitive to acute
insulin administration [1–3,5,18] and highlights the phenotypic
difference between life-long genetic myostatin blockade and adult
age treatment with myostatin antagonists. Results Effects of D76A on Body Composition August 2013 | Volume 8 | Issue 8 | e71017 Effects of D76A on Aortic Atherosclerosis The atheromatous lesions in the aorta were analyzed using the
en face method [5,12]. Most of the lesions were found in the aortic
arch and fewer lesions were present in the thoracic and abdominal
regions. D76A-treated mice had significantly fewer lesions than
vehicle-treated mice (Figure 1A, additional images of aorta lesion
staining are displayed in Figure S3). The mean lesion scores in the
aortic arch and the thoracic aorta were 57% and 83% lower in
D76A-treated animals than those in the control mice, respectively
(Figure 1A, lower panel). Hematoxylin and eosin staining of the
sections near the sinus of the aortic root revealed a significant
reduction of lesion size in D76A-treated mice compared with the
control mice (Figure 1B), consistent with the en face assessment. Similar results were observed by trichrome staining of the aortic
roots, which further revealed a decrease in lipid core size and the
cholesterol clefts in mice treated with D76A (Figure S4). To assess the inflammatory status of the lesions, we performed
immunostaining of the aortic root sections for MAC3, a marker of
macrophage infiltration. As shown in Figure 2A, vehicle-treated
mice showed greater MAC3 staining than D76A-treated mice. At
higher magnification, the lesions from D76A-treated mice were
found to contain fewer and less intensely stained MAC3-positive
cells than those from vehicle-treated mice (Figure 2A). Densito-
metric analysis revealed 41% reduction in MAC3 staining in
AAV-D76A-treated mice compared to controls (Figure 2A, lower
panel). As endothelial inflammation usually begins with the Unlike muscle and fat that take up glucose in response to
insulin, liver produces glucose in response to nutrient deprivation,
a process that is potently inhibited by insulin [21]. An increase in
basal (non-fasting) plasma glucose concentration can be a sign of
impaired insulin sensitivity in the liver so that it continues to 3 PLOS ONE | www.plosone.org August 2013 | Volume 8 | Issue 8 | e71017 3 Myostatin, Hepatosteatosis, and Atherosclerosis Figure 1. AAV-mediated expression of myostatin pro-peptide D76A mutant attenuates aortic atheromatous lesion accumulation in
Ldlr null mice. The animals were injected with either AAV-D76A (D76A) or vehicle (veh) at 8 weeks of age, placed on a high fat diet seven weeks
later, and euthanized after another 12 weeks. The entire aorta along with the aortic root was dissected for analysis. (A) En face staining with Sudan IV
for lipid-rich lesions (stained red). Effects of D76A on Aortic Atherosclerosis The lower panel shows the quantitative lesion scores (mean 6 SEM, n = 10). (B) Sections of the aortic root near the
sinus stained with hematoxylin and eosin (H&E) for lesions built up along the vessel wall (LC: lipid core, VW: vessel wall). The lower panel shows the
lesion area expressed as a percent of aortic area (mean6SEM, n = 10). doi:10.1371/journal.pone.0071017.g001 Figure 1. AAV-mediated expression of myostatin pro-peptide D76A mutant attenuates aortic atheromatous lesion accumulation in
Ldlr null mice. The animals were injected with either AAV-D76A (D76A) or vehicle (veh) at 8 weeks of age, placed on a high fat diet seven weeks
later, and euthanized after another 12 weeks. The entire aorta along with the aortic root was dissected for analysis. (A) En face staining with Sudan IV
for lipid-rich lesions (stained red). The lower panel shows the quantitative lesion scores (mean 6 SEM, n = 10). (B) Sections of the aortic root near the
sinus stained with hematoxylin and eosin (H&E) for lesions built up along the vessel wall (LC: lipid core, VW: vessel wall). The lower panel shows the
lesion area expressed as a percent of aortic area (mean6SEM, n = 10). doi:10.1371/journal.pone.0071017.g001 levels of insulin receptor substrate 2 (IRS-2, Figure 3C) but not
IRS-1 (not shown). Of note, IRS-2 has been reported to play a
more important role than IRS-1 in hepatic insulin action [22]. Consistently, D76A-treated animals also displayed lower liver
expression of Pepck (Figure 3C), a key enzyme for gluconeogenesis
known to be transcriptionally suppressed by insulin [5,23]. Together, these results suggest that D76A-treated animals have produce glucose even when nutrient is plentiful (Figure 4B, left
panel). Hence, we performed the pyruvate tolerance test, which
measures the rise of blood glucose in response to a bolus injection
of pyruvate. As shown in Figure 3B (right panel), blood glucose
concentrations rose faster and to a higher level in the control mice
than in D76A-treated mice, suggesting that the latter had a greater
restraint on hepatic glucose output. Real-time PCR analysis
showed that D76A-treated mice had higher hepatic expression Figure 2. AAV-D76A reduces expression of MAC-3 and VCAM-1 in aortic lesions. Effects of D76A on Aortic Atherosclerosis Sections adjacent to the sinus of the aortic roots were
immuno-stained using antibodies against (A) CD107b (MAC-3), a marker of macrophage infiltration, and (B) vascular cell adhesion molecule (VCAM-
1), an endothelial ligand that promotes the adhesion of lymphocytes, monocytes, eosinophils, and basophils (EC: endothelial cells, VW: vessel wall, LC:
lipid core). Lower panels display the quantitative analyses of immunostaining determined by densitometry (mean6SEM, n = 5). doi:10.1371/journal.pone.0071017.g002 Figure 2. AAV-D76A reduces expression of MAC-3 and VCAM-1 in aortic lesions. Sections adjacent to the sinus of the aortic roots were
immuno-stained using antibodies against (A) CD107b (MAC-3), a marker of macrophage infiltration, and (B) vascular cell adhesion molecule (VCAM-
1), an endothelial ligand that promotes the adhesion of lymphocytes, monocytes, eosinophils, and basophils (EC: endothelial cells, VW: vessel wall, LC:
lipid core). Lower panels display the quantitative analyses of immunostaining determined by densitometry (mean6SEM, n = 5). doi:10.1371/journal.pone.0071017.g002 August 2013 | Volume 8 | Issue 8 | e71017 PLOS ONE | www.plosone.org 4 Myostatin, Hepatosteatosis, and Atherosclerosis Figure 3. AAV-D76A regulates plasma lipid profile and insulin sensitivity. (A) Plasma lipid profile: fasting plasma samples were obtained
after 10 weeks of high fat feeding and analyzed for triglycerides (TG), total cholesterol (Total C), HDL cholesterol (HDL-C), non HDL cholesterol
(nonHDL-C), and non-esterified fatty acids (NEFA). (B) Insulin tolerance test (ITT) and pyruvate tolerance test (PTT) performed after 11 weeks of high
fat feeding. For ITT, food was removed in the morning for four hours and then animals were injected with an insulin bolus (0.6 U/kg diluted in sterile
saline containing 0.1% albumin, i.p.). For PTT, food was removed overnight for 16 hours and then animals were injected sodium pyruvate (1.5 g/kg,
i.p. diluted in sterile saline). Blood glucose was measured using a glucometer and was plotted against time before and after the injections (n = 10,
mean 6 SEM). (C) Liver mRNA expression of insulin receptor substrate-2 (IRS-2) and phosphoenolpyruvate carboxykinase (Pepck) was analyzed by
real-time PCR (mean6SEM, n = 10). doi:10.1371/journal.pone.0071017.g003 Figure 3. AAV-D76A regulates plasma lipid profile and insulin sensitivity. (A) Plasma lipid profile: fasting plasma samples were obtained
after 10 weeks of high fat feeding and analyzed for triglycerides (TG), total cholesterol (Total C), HDL cholesterol (HDL-C), non HDL cholesterol
(nonHDL-C), and non-esterified fatty acids (NEFA). (B) Insulin tolerance test (ITT) and pyruvate tolerance test (PTT) performed after 11 weeks of high
fat feeding. Effects of D76A on Aortic Atherosclerosis For ITT, food was removed in the morning for four hours and then animals were injected with an insulin bolus (0.6 U/kg diluted in sterile
saline containing 0.1% albumin, i.p.). For PTT, food was removed overnight for 16 hours and then animals were injected sodium pyruvate (1.5 g/kg,
i.p. diluted in sterile saline). Blood glucose was measured using a glucometer and was plotted against time before and after the injections (n = 10,
mean 6 SEM). (C) Liver mRNA expression of insulin receptor substrate-2 (IRS-2) and phosphoenolpyruvate carboxykinase (Pepck) was analyzed by
real-time PCR (mean6SEM, n = 10). doi:10.1371/journal.pone.0071017.g003 Figure 4. AAV-D76A attenuates liver fat infiltration, reduces liver triglycerides and cholesterol, and down-regulates mRNA
expression of selected lipogenic genes. (A) Cryo-sectioned liver samples (10 m) were stained with oil-red-O (red for lipids). Results are
representative of three animals per group. (B) Liver lipids were extracted by Folch’s method and analyzed for total cholesterol (Total C) and
triglycerides (TG) using commercial kits (mean 6 SEM, n = 10). (C) Liver mRNA expression was analyzed using qPCR; the upper panel shows genes
involved in lipid synthesis and the lower panel shows the genes involved in lipid oxidation (FAS: fatty acid synthase, SCD-1: stearoyl-CoA desaturase-
1, HMGCR: 3-hydroxy-3-methyl-glutaryl-CoA reductase, SREBP-1c: sterol regulatory element-binding protein 1, PPAR: peroxisome proliferator-
activated receptor, CPT: carnitine palmitoyl acyltransferase, MACD: medium chain acylCoA dehydrogenase). All PCR results were normalized to the
expression level of the house-keeping gene HPRT (mean 6 SEM, n = 10). doi:10.1371/journal.pone.0071017.g004 Figure 4. AAV-D76A attenuates liver fat infiltration, reduces liver triglycerides and cholesterol, and down-regulates mRNA
expression of selected lipogenic genes. (A) Cryo-sectioned liver samples (10 m) were stained with oil-red-O (red for lipids). Results are
representative of three animals per group. (B) Liver lipids were extracted by Folch’s method and analyzed for total cholesterol (Total C) and
triglycerides (TG) using commercial kits (mean 6 SEM, n = 10). (C) Liver mRNA expression was analyzed using qPCR; the upper panel shows genes
involved in lipid synthesis and the lower panel shows the genes involved in lipid oxidation (FAS: fatty acid synthase, SCD-1: stearoyl-CoA desaturase-
1, HMGCR: 3-hydroxy-3-methyl-glutaryl-CoA reductase, SREBP-1c: sterol regulatory element-binding protein 1, PPAR: peroxisome proliferator-
activated receptor, CPT: carnitine palmitoyl acyltransferase, MACD: medium chain acylCoA dehydrogenase). All PCR results were normalized to the
expression level of the house-keeping gene HPRT (mean 6 SEM, n = 10). Liver is a Novel Direct Target of Myostatin Because the atheroprotective phenotype mediated by AAV-
D76A injection was only weakly associated with changes in muscle
mass, but strongly associated with changes in hepatic fat and
robust hepatic expression of ectopic D76A, we considered the
possibility that metabolic protection may be a result of direct
blockade of myostatin signaling in the liver. To our knowledge,
direct effect of myostatin on liver has not been reported although
several studies have speculated such a possibility [3,29,30]. There
has been some uncertainty whether ActRIIB, the high affinity
receptor for myostatin [31,32], is expressed in the liver [33–36]. Using RT-PCR, we confirmed that ActRIIB was expressed in the
mouse liver although to a moderate extent (Figure 5A). When
recombinant myostatin was added to liver cells (HepG2) in culture,
ActRIIB
expression
was
induced
in
a
sustained
manner
(Figure 5B). Similar induction of ActRIIB was confirmed in
isolated primary mouse hepatocytes (Figure S6). Myostatin rapidly
induced Smad3 phosphoylation (Figure 5C), an essential compo-
nent in the canonical myostatin signaling cascade [37,38]. Myostatin
also
increased
the
expression
of
selected
genes
downstream of Smad3; including TGFb1, apoCIII, PAI-1, and
angiotensinogen II (AngII) (Figure 5D). PAI-1 and apoCIII are
known to be transcriptionally regulated by Smad3 [39,40],
whereas AngII can be induced by TGFb1 via a Smad-
independent pathway [41]. In line with these in vitro findings,
we show that inhibition of myostatin by D76A was associated with
decreased expression of TGFb1 and apoCIII in the liver (Figure
S5, lower panel). In addition, cells treated with myostatin also
showed increased expression of enzymes involved in lipogenesis,
such as SCD-1 and FAS (Figure 5D, lower panel). Consistent with
the increase in the expression of lipogenic genes, we found that Effects of D76A on Aortic Atherosclerosis doi:10.1371/journal.pone.0071017.g004 August 2013 | Volume 8 | Issue 8 | e71017 5 PLOS ONE | www.plosone.org 5 Myostatin, Hepatosteatosis, and Atherosclerosis Figure 5. Liver is a direct target of myostatin signaling. (A) PCR measurement of the expression of myostatin receptor, ActRIIB, in the liver,
aorta, and skeletal muscle. (B) Myostatin induces sustained expression of its own receptor in HepG2 cells (mstn, 100 ng/ml, harvested at indicated
time points). (C) Myostatin induces Smad3 phosphorylation in HepG2 cells (100 ng/ml, 30 min). All results shown in (A–C) are representative of more
than three independent measurements. (D) Myostatin (100 ng/ml, 24 h) induces expression of Smad3 target genes in HepG2 cells (mean 6 SEM,
n = 4). (E) Myostatin (100 ng/ml) induces lipogenesis and lipid accumulation in HepG2 cells (means 6 SEM, n = 4). doi:10.1371/journal.pone.0071017.g005 Figure 5. Liver is a direct target of myostatin signaling. (A) PCR measurement of the expression of myostatin receptor, ActRIIB, in the liver,
aorta, and skeletal muscle. (B) Myostatin induces sustained expression of its own receptor in HepG2 cells (mstn, 100 ng/ml, harvested at indicated
time points). (C) Myostatin induces Smad3 phosphorylation in HepG2 cells (100 ng/ml, 30 min). All results shown in (A–C) are representative of more
than three independent measurements. (D) Myostatin (100 ng/ml, 24 h) induces expression of Smad3 target genes in HepG2 cells (mean 6 SEM,
n = 4). (E) Myostatin (100 ng/ml) induces lipogenesis and lipid accumulation in HepG2 cells (means 6 SEM, n = 4). doi:10.1371/journal.pone.0071017.g005 improved hepatic insulin sensitivity in comparison to vehicle-
treated mice under our experimental conditions. Effects of D76A on Hepatosteatosis and Hepatic
Metabolic Gene Expression Fatty liver is linked to diet-induced dyslipidemia and athero-
sclerosis [24]. We found that D76A-treated mice had significantly
lower liver fat than the control mice, as indicated by lipid staining
of liver sections (Figure 4A) and by lipid analysis of the liver
extracts (Figure 4B). D76A-treated mice also had lower hepatic
mRNA expression of key enzymes involved in lipogenesis,
including fatty acid synthase (FAS), stearoyl CoA dehydrogenase
(SCD-1), and HMG CoA reductase (HMGCR) (Figure 4C, upper
panel). These enzymes are largely regulated at the transcriptional
level [25]. Consistently, we found that liver expression level of
SREBP-1c, the master transcription regulator of hepatic lipogenic
enzymes [26,27], was lower in D76A-treated animals than in the
controls. In contrast to its inhibitory effect on lipogenic gene
expression, D76A treatment had no effect on hepatic expression of
important genes involved in fatty acid oxidation (Figure 4C, lower
panel). D76A treatment also did not affect the expression of
selected inflammatory cytokines, such as TNFa, MCP-1, and CRP
(Figure S4). Plasma IL-6 and MCP-1 were very low and were not
significantly affected by D76A (data not shown). The expression
levels of selected stress markers, C/EBP homologous protein-10
(CHOP-10) and heme oxygenase 1 (HO-1), were higher in the
liver of control mice than D76A-treated mice (Figure S5),
consistent with the notion that fat infiltration induces expression
of stress genes [28]. PLOS ONE | www.plosone.org August 2013 | Volume 8 | Issue 8 | e71017 August 2013 | Volume 8 | Issue 8 | e71017 6 Myostatin, Hepatosteatosis, and Atherosclerosis myostatin increased fatty acid synthesis and lipid accumulation in
cultured liver cells (Figure 5E). There was no effect of myostatin on
fatty acid oxidation measured in parallel (data not shown). To our
knowledge, this is the first report of direct pro-lipogenic effect of
myostatin on liver cells, although others have shown that TGFb1,
which shares similar post-receptor signaling pathways with
myostatin, increases SCD-1 expression and increases lipid storage
in cultured liver cells [42,43]. to cause global metabolic protections [1,3,18]. However, such
extremely hypertrophic muscle phenotype was not reproduced in
mice with adult age myostatin inhibition [6–9,19,47](this work). In
addition, life-long genetic myostatin blockade, global or muscle-
specific, prevents diet-induced increment in fat mass [1–3,5], a
phenotype that was also not reproducible in mice with adult age
myostatin inhibition ([19], this work). Aortic Endothelium can be another Direct Target of
Myostatin y
As shown in Figure 5A, the high affinity myostatin receptor,
ActRIIB, was robustly expressed in mouse aorta, so was
endogenous myostatin gene (Figure 6A). Within the aortic
endothelium, endothelial cells represent the first line of defense
against the insult from circulating factors. We thus evaluated the
direct effects of myostatin on aortic endothelial cells. Similar to
that found in the liver cells (Figure 5C), myostatin induced robust
Smad3 phosphorylation in both bovine and primary human aortic
cells (not shown). Figure 6B demonstrated that myostatin directly
induced activation of 3TP-Lux reporter gene for TGFb signaling
in
bovine
aortic
endothelial
cells,
an
effect
that
requires
cooperation between both type-II and type-I receptors [10]. Since
type-I receptors are shared between myostatin and TGFb [44], the
induction of 3TP-Lux by myostatin confirms functional activation
of myostatin signaling in the aortic endothelial cells. Furthermore,
incubation with myostatin induced the expression of intracellular
cell adhesion molecule 1 (ICAM-1, Figure 6C), down-regulated
steady-state phosphorylation/activation of endothelial NO syn-
thase (eNOS, Figure 6D) and up-regulated the expression of
vascular cell adhesion molecule (VCAM-1, Figure 6D) in primary
cultured human aortic endothelial cells (HuAEC). As eNOS
protects the endothelium against insults from circulating inflam-
matory cytokines and oxidized lipoproteins [45] while ICAM-1
and VCAM-1 promote monocyte adhesion and lesion develop-
ment [46], these results suggest that myostatin blockade may have
a favorable effect on the endothelium under certain physiological
conditions, such as hyperlipidemia, by reducing local expression of
pro-adhesion
molecules,
a
hypothesis
that
requires
further
investigations. Although protection against diet-induced hepatosteatosis has
been documented in mice with genetic myostatin blockade for life
or induced at adult age [1–3,5,19], it is generally considered as
secondary to the impact on muscle health [1,30]. To our
knowledge, the results provided in this work are the first evidence
that myostatin can participate in hepatic metabolic regulation
through direct interaction with liver cells. We show that liver cells
express high affinity receptor for myostatin and respond to
myostatin stimulation by increasing phosphorylation of Smad3
and expression of Smad3-responsive genes including SCD-1 [42],
a key lipogenic gene in fatty liver development [48]. Furthermore,
we have shown that myostatin induces the expression of selected
lipogenic genes and increases lipid accumulation in the cultured
liver cells. This finding is congruent with the reports that TGFb1,
which shares the type I receptor with myostatin, induces lipogenic
gene expression and increases lipid accumulation in liver cells
[42,43]. Aortic Endothelium can be another Direct Target of
Myostatin A second novel finding reported in this work is the direct effect
of myostatin on aortic endothelial cells. While liver plays a central
role in systemic lipid metabolism, aortic atherosclerosis starts with
endothelial damage and monocyte adherence, which set the stage
for lipid uptake and foam cell formation. We show here that
myostatin and its receptor are robustly expressed in the aortic
tissue. In addition, we provide evidence that myostatin directly
interacts with cultured bovine aortic endothelial cell line and
primary human aortic endothelial cells to inhibit eNOS activation
and up-regulate the expression of ICAM-1 and VCAM-1. These
effects potentially could contribute to arterial lesion development
and warrants further evaluations. Effects of D76A on Hepatosteatosis and Hepatic
Metabolic Gene Expression However, despite a modest
impact on body composition, adult age myostatin inhibition does
prove effective in protection against diet-induced fatty liver ([19],
this work) and atherosclerosis (this work), thus prompting us to
consider organ-specific effect of myostatin (and its inhibitors) on
metabolic regulations without being overwhelmed by the hyper-
muscular phenotype found in mice with life-long genetic myostatin
blockade. Discussion Indeed, it has been shown that life-long muscle-
specific transgenic myostatin blockade was sufficient to reproduce
the phenotype of mice with global myostatin knockout, unequiv-
ocally proving that dramatic muscle hypertrophy alone is sufficient August 2013 | Volume 8 | Issue 8 | e71017 August 2013 | Volume 8 | Issue 8 | e71017 7 PLOS ONE | www.plosone.org Myostatin, Hepatosteatosis, and Atherosclerosis Figure 6. Aorta is a direct target of myostatin signaling. (A) PCR analysis of aortic expression of endogenous myostain mature peptide (mstn,
nt 1015–1145, NM_010834), using skeletal muscle as the positive control and liver as the negative control (mean 6 SEM, n = 3). Of note, while actin is
often considered as an abundant house-keeping gene in multiple tissues, its expression level may not be cross-comparable. Hence, the results
presented here only demonstrated that myostatin is abundantly expressed in the aorta but does not indicate that the expression is to a similar level
found in the muscle. (B) Myostatin induces activation of its reporter gene 3TP-Lux in bovine aortic endothelial cells (BAEC, mean 6 SEM, n = 3). (C)
Myostain induces ICAM-1 expression in primary cultured human aortic endothelial cells (HuAEC). TGFb1 (T: 10 ng/ml) served as a positive control
(M50: myostatin 50 ng/ml, M100:100 ng/ml, 3 h incubation in serum-free Medium 199 supplemented with oxLDL 0.025 mg/ml to raise the basal
expression of ICAM-1) (mean 6 SEM, n = 4). (D) Myostatin (100 ng/ml) down regulates the expression of phosphor-eNOS and up regulates VCAM-1
expression in HuAEC. Results are reflective of three independent experiments. doi:10.1371/journal.pone.0071017.g006 Figure 6. Aorta is a direct target of myostatin signaling. (A) PCR analysis of aortic expression of endogenous myostain mature peptide (mstn,
nt 1015–1145, NM_010834), using skeletal muscle as the positive control and liver as the negative control (mean 6 SEM, n = 3). Of note, while actin is
often considered as an abundant house-keeping gene in multiple tissues, its expression level may not be cross-comparable. Hence, the results
presented here only demonstrated that myostatin is abundantly expressed in the aorta but does not indicate that the expression is to a similar level
found in the muscle. (B) Myostatin induces activation of its reporter gene 3TP-Lux in bovine aortic endothelial cells (BAEC, mean 6 SEM, n = 3). (C)
Myostain induces ICAM-1 expression in primary cultured human aortic endothelial cells (HuAEC). (TIF) Figure S3
Effects of AAV-D76A on aorta lesion accumulation:
additional en face aortic images supplemental to Figure 1. The
microphotographs were taken on a different facility and more
yellowish background color was recorded. Lipid-rich lesions are
stained red with Sudan IV. The results clearly illustrated a
reduction of aortic lesions in D76A-treated mice compared to the
vehicle-treated ones. Discussion It should be pointed out that while diet-induced hyperlipidemia
is an established risk factor for atherosclerosis, the effect of D76A
on plasma lipid profile in our animals was relatively moderate,
which alone may not explain the marked reduction in aortic
lesions. Other mechanisms can be involved. For instance, liver
secretion of D76A may help to sequester circulating myostatin in
its inactive form; reducing its impact on endothelial production of
adhesive molecules. In addition to regulation of liver lipogenesis,
myostatin blockade by D76A may also change some of the
properties of liver secretion, such as the size, composition, and
redox state of the VLDL particles, which can result in less pro-
atherogenic lipid particles in circulation. Furthermore, excessive
lipid accumulation is often associated with tissue inflammation. Although D76A did not change the expression of selected
common inflammatory markers in the liver (Figure S5) and in
circulation (data not shown), we show that D76A reduced liver
expression of TGFb1 and apoCIII, both can be pro-atherogenic
[49,50]. D76A also reduced liver expression of stress marker
CHOP-10 and HO-1, implying less oxidative damage in the liver
which can alter the antioxidant defense in secreted lipid particles
as well as other systemic redox regulators. Further studies are Our data provide the first evidence that AAV-mediated
administration of myostatin pro-peptide D76A mutant in adult
mice attenuates diet-induced aortic atherosclerosis and hepatos-
teatosis in Ldlr null mice. These data provide a new rationale to
support the development of anti-myostatin strategies for the
prevention and treatment of atherosclerosis and hepatosteatosis in
humans. It is important to emphasize that although genetic modulation is
useful to validate a concept, the feasibility for translational
medicine is entirely dependent on whether a favorable phenotype
of a genetic model can be successfully recapitulated by post-natal,
especially adult age, interventions. Life-long genetic myostatin
blockade causes dramatic hyper-muscularity and hypo-adiposity
[1–3,5]. Hence, it is not clear whether the associated metabolic
phenotype, i.e. strong protection against diet-induced insulin
resistance, hepatosteatosis, and atherosclerosis [1–3,5], is directly
related to myostatin signaling or secondary to the extreme muscle
hypertrophy. Discussion TGFb1 (T: 10 ng/ml) served as a positive control
(M50: myostatin 50 ng/ml, M100:100 ng/ml, 3 h incubation in serum-free Medium 199 supplemented with oxLDL 0.025 mg/ml to raise the basal
expression of ICAM-1) (mean 6 SEM, n = 4). (D) Myostatin (100 ng/ml) down regulates the expression of phosphor-eNOS and up regulates VCAM-1
expression in HuAEC. Results are reflective of three independent experiments. doi:10.1371/journal.pone.0071017.g006 (TIF) required to investigate these and other possible mechanisms to
fully understand the remarkable phenotype observed in this study. fully understand the remarkable phenotype observed in this study. The pioneering work of Lee and others have emphasized the
important evolutionary role of myostatin as a chalone that
restrains unbridled muscle growth [31]. The findings of the
current study suggest that myostatin may also promote hepatic fat
stores and facilitate vessel wall wound healing; both can be
important pro-survival evolutionary adaptations during periods of
nutrient paucity. However, in the contemporary period of nutrient
excess coupled with physical inactivity, the same function of
myostatin could contribute to metabolic dys-regulations [51–55]. Figure S2
Effects of myostatin inhibition by AAV-D76A on
body composition. A: Time-dependent changes in body lean mass
after injection of AAV-D76A during the first 7 weeks while the
animals were fed normal chow. B: The net increase in total lean
(left) and fat (right) mass during the first seven weeks (left panel),
the 12 week of high fat diet (middle panel), and total experimental
period (right panel). Results of A&B were obtained by NMR
(mean +/2 SE, N = 10, *p,0.05). C: Tissue weight expressed as
percentage of total body mass (means +/2 se, n = 10). Mean body
mass was not significantly different between the two groups
(43.29+/22.96 g and 41.49+/23.62 g, for vehicle- and D76A-
treated groups, respectively, n = 10). (TIF) Figure S2
Effects of myostatin inhibition by AAV-D76A on
body composition. A: Time-dependent changes in body lean mass
after injection of AAV-D76A during the first 7 weeks while the
animals were fed normal chow. B: The net increase in total lean
(left) and fat (right) mass during the first seven weeks (left panel),
the 12 week of high fat diet (middle panel), and total experimental
period (right panel). Results of A&B were obtained by NMR
(mean +/2 SE, N = 10, *p,0.05). C: Tissue weight expressed as
percentage of total body mass (means +/2 se, n = 10). Mean body
mass was not significantly different between the two groups
(43.29+/22.96 g and 41.49+/23.62 g, for vehicle- and D76A-
treated groups, respectively, n = 10). (TIF) (TIF) y
y
g
In summary, this work has demonstrated that (i) myostatin may
participate in signal regulation and modulate metabolic outcomes
through direct interactions with target cells in the liver and the
aorta; (ii) myostatin antagonists might have a therapeutic role in
the prevention or treatment of hepatosteatosis and atherosclerosis. These findings set the stage for further investigations of the
potential therapeutic applications of myostatin antagonists for
metabolic disorders as well as the mechanisms by which myostatin
signaling
pathways
intersect
with
the
metabolic
regulatory
pathways. References 18. Guo T, Bond ND, Jou W, Gavrilova O, Portas J, et al. (2012) Myostatin
inhibition prevents diabetes and hyperphagia in a mouse model of lipodystrophy. Diabetes. 61: 2414–2423. 1. Guo T, Jou W, Chanturiya T, Portas J, Gavrilova O, et al. (2009) Myostatin
inhibition in muscle, but not adipose tissue, decreases fat mass and improves
insulin sensitivity. PLoS One. 4: e4937. 1. Guo T, Jou W, Chanturiya T, Portas J, Gavrilova O, et al. (2009) Myostatin
inhibition in muscle, but not adipose tissue, decreases fat mass and improves
insulin sensitivity. PLoS One. 4: e4937. y
2. Wilkes JJ, Lloyd DJ, Gekakis N (2009) Loss-of-function mutation in myostatin
reduces tumor necrosis factor alpha production and protects liver against
obesity-induced insulin resistance. Diabetes. 58: 1133–1143. 19. Burgess K, Xu T, Brown R, Han B, Welle S (2011) Effect of myostatin depletion
on weight gain, hyperglycemia, and hepatic steatosis during five months of high-
fat feeding in mice. PLoS One. 6: e17090. 3. McPherron AC, Lee SJ (2002) Suppression of body fat accumulation in
myostatin-deficient mice. J Clin Invest. 109: 595–601. 20. Dashti N, Wolfbauer G (1987) Secretion of lipids, apolipoproteins, and
lipoproteins by human hepatoma cell line, HepG2: effects of oleic acid and
insulin. J Lipid Res. 28: 423–436. y
J
4. McPherron AC, Lawler AM, Lee SJ (1997) Regulation of skeletal muscle mass in
mice by a new TGF-beta superfamily member. Nature. 387: 83–90. 21. Choudhury J, Sanyal AJ (2005) Insulin resistance in NASH. Front Biosci. 10:
1520–1533. 5. Tu P, Bhasin S, Hruz PW, Herbst KL, Castellani LW, et al. (2009) Genetic
disruption of myostatin reduces the development of proatherogenic dyslipidemia
and atherogenic lesions in Ldlr null mice. Diabetes. 58: 1739–1748. 22. Kohjima M, Higuchi N, Kato M, Kotoh K, Yoshimoto T, et al. (2008) SREBP-
1c, regulated by the insulin and AMPK signaling pathways, plays a role in
nonalcoholic fatty liver disease. Int J Mol Med. 21: 507–511. 6. Wolfman NM, McPherron AC, Pappano WN, Davies MV, Song K, et al (2003)
Activation of latent myostatin by the BMP-1/tolloid family of metalloprotei-
nases. Proc Natl Acad Sci USA. 100: 15842–15846. 23. Srivastava RA, Jahagirdar R, Azhar S, Sharma S, Bisgaier CL (2006)
Peroxisome proliferator-activated receptor-alpha selective ligand reduces
adiposity, improves insulin sensitivity and inhibits atherosclerosis in LDL
receptor-deficient mice. Mol Cell Biochem. 285: 35–50. 7. Foster K, Graham IR, Otto A, Foster H, Trollet C, et al. References (2009) Adeno-
associated virus-8-mediated intravenous transfer of myostatin propeptide leads
to systemic functional improvements of slow but not fast muscle. Rejuvenation
Res. 12: 85–94. 24. Bugianesi E, Moscatiello S, Ciaravella MF, Marchesini G (2010) Insulin
resistance in nonalcoholic fatty liver disease. Curr Pharm Des. 16: 1941–1951. y
25. Pallottini V, Guantario B, Martini C, Totta P, Filippi I, et al. (2008) Regulation
of HMG-CoA reductase expression by hypoxia. J Cell Biochem. 104: 701–709. 8. Qiao C, Li J, Jiang J, Zhu X, Wang B, et al. (2008) Myostatin propeptide gene
delivery by adeno-associated virus serotype 8 vectors enhances muscle growth
and ameliorates dystrophic phenotypes in mdx mice. Hum Gene Ther. 19: 241–
254. 26. Kaplan M, Kerry R, Aviram M, Hayek T (2008) High glucose concentration
increases macrophage cholesterol biosynthesis in diabetes through activation of
the sterol regulatory element binding protein 1 (SREBP1): inhibitory effect of
insulin. J Cardiovasc Pharmacol. 52: 324–332. 9. Morine KJ, Bish LT, Pendrak K, Sleeper MM, Barton ER, et al (2010) Systemic
myostatin inhibition via liver-targeted gene transfer in normal and dystrophic
mice. PLoS One. 5: e9176. 27. Kotzka J, Knebel B, Avci H, Jacob S, Nitzgen U, et al. (2010) Phosphorylation of
sterol regulatory element-binding protein (SREBP)-1a links growth hormone
action to lipid metabolism in hepatocytes. Atherosclerosis. 213: 156–165. 10. Wrana JL, Attisano L, Carcamo J, Zentella A, Doody J, et al. (1992) TGF beta
signals through a heteromeric protein kinase receptor complex. Cell. 71: 1003–
1014. 28. Gentile CL, Frye M, Pagliassotti MJ (2011) Endoplasmic reticulum stress and the
unfolded protein response in nonalcoholic fatty liver disease. Antioxid Redox
Signal. 15: 505–521. 11. Li D, Liu Y, Chen J, Velchala N, Amani F, et al (2006) Suppression of
atherogenesis by delivery of TGFbeta1ACT using adeno-associated virus type 2
in LDLR knockout mice. Biochem Biophys Res Commun. 344: 701–707. g
29. McPherron AC (2010) Metabolic functions of myostatin and GDF11. Immunol
Endocr Metab Agents Med Chem. 10: 217–231. 12. Guo W, Wong S, Pudney J, Jasuja R, Hua N, et al (2009) Acipimox, an inhibitor
of lipolysis, attenuates atherogenesis in LDLR-null mice treated with HIV
protease inhibitor ritonavir. Arterioscler Thromb Vasc Biol. 29: 2028–2032. 30. Allen DL, Hittel DS, McPherron AC (2011) Expression and function of
myostatin in obesity, diabetes, and exercise adaptation. Med Sci Sports Exerc. 43: 1828–1835. p
13. Acknowledgments We thank Amgen for providing recombinant myostatin peptide, Dr. Se-Jin
Lee (Johns Hopkins University) for the cDNA clone of the myostatin
propeptide mutant, and Drs. Mengwei Zang and Dr. Yasuo Ito (Boston
University) for the HepG2 and BAEC cell lines. We thank Dr. Laura
Nocito (Boston University) for the help with primary hepatocyte isolation
and we thank Dr. Shoupu Chen (Carestream) for the help with image
processing. Figure S6
Primary mouse hepatocytes were isolated by the
portal vein collagenase perfusion method as described (Li WC,
Ralphs KL, Tosh D. Isolation and culture of adult mouse
hepatocytes. Methods
Mol
Biol. 2010;633:185–96. PMID:
20204628). After attachment, medium was replaced by serum-
free M199 overnight. Myostatin (mstn) was added the next
morning and incubated for 5 hours before RNA harvest. Expression of ActRIIB (NM_007397) was measured by qPCR (f- Figure S6
Primary mouse hepatocytes were isolated by the
portal vein collagenase perfusion method as described (Li WC,
Ralphs KL, Tosh D. Isolation and culture of adult mouse
hepatocytes. Methods
Mol
Biol. 2010;633:185–96. PMID:
20204628). After attachment, medium was replaced by serum-
free M199 overnight. Myostatin (mstn) was added the next
morning and incubated for 5 hours before RNA harvest. Expression of ActRIIB (NM_007397) was measured by qPCR (f- primer:
CATTGCTGCCGAGAAACGAG;
r-primer:
TCCACGTGATGATGTTCCCC ). Exogenous myostatin in-
duces expression of its own receptor ActRIIB in a bell-shape like
dose-dependent manner.
(TIF) primer:
CATTGCTGCCGAGAAACGAG;
r-primer:
TCCACGTGATGATGTTCCCC ). Exogenous myostatin in-
duces expression of its own receptor ActRIIB in a bell-shape like
dose-dependent manner. (TIF) Figure S5
Changes in liver transcripts after myostatin inhibition
by AAV-D76A. Liver mRNA was analyzed by RT-qPCR for
expression of genes downstream of TGFb/mystatin signaling:
TGFb1, ApoCIII, ANG-II and PAI-I (lower panel), stress-related
genes: BIP, CHOP, and HO-1 (upper panel), and inflammatory
genes: MCP-1, TNFa, and CRP (middle panel). Results are shown
as means +/2 se, n = 10, t test). (TIF) Author Contributions Conceived and designed the experiments: WG SB. Performed the
experiments: SW WG. Analyzed the data: WG SW SB. Wrote the paper:
WG SW SB. Conceived and designed the experiments: WG SB. Performed the
experiments: SW WG. Analyzed the data: WG SW SB. Wrote the paper:
WG SW SB. (TIF) Figure S4
Trichrome staining of lesions from the vehicle and
D76A- treated mice. Red stains for muscle, including the smooth
muscle in the fibrous cap at the lumen side, blue stains for
collagen, white foamy structure indicates lipid-rich foam cell Figure S1
Expression of myostatin propeptide (nt 222–214,
NM_010834) in the liver (left panel) and quadriceps muscle (right
panel) between vehicle and D76A-treated animals, normalized to
the expression of house-keeping gene HPRT. Each bar represents
one individual animal. August 2013 | Volume 8 | Issue 8 | e71017 PLOS ONE | www.plosone.org 8 Myostatin, Hepatosteatosis, and Atherosclerosis deposits, needle-like structures are cholesterol clefts from necrotic
lipid core. Results are representative of five animals of each group. (TIF) Myostatin, Hepatosteatosis, and Atherosclerosis 36. Gold EJ, Francis RJ, Zimmermann A, Mellor SL, Cranfield M, et al. (2003)
Changes in activin and activin receptor subunit expression in rat liver during the
development of CCl4-induced cirrhosis. Mol Cell Endocrinol. 201: 143–153. adhesion molecule 1 via the Akt and nuclear factor-kappaB-dependent pathway,
leading to suppression of adhesion of monocytes and eosinophils to bronchial
epithelial cells. Immunology. 137: 98–113. adhesion molecule 1 via the Akt and nuclear factor-kappaB-dependent pathway,
leading to suppression of adhesion of monocytes and eosinophils to bronchial
epithelial cells. Immunology. 137: 98–113. 37. Guo W, Flanagan J, Jasuja R, Kirkland J, Jiang L,et al. (2008) The effects of
myostatin on adipogenic differentiation of human bone marrow-derived
mesenchymal stem cells are mediated through cross-communication between
Smad3 and Wnt/beta-catenin signaling pathways. J Biol Chem. 283: 9136–
9145. 47. LeBrasseur NK, Schelhorn TM, Bernardo BL, Cosgrove PG, Loria PM, et al. (2009) Myostatin inhibition enhances the effects of exercise on performance and
metabolic outcomes in aged mice. J Gerontol A Biol Sci Med Sci. 64: 940–948. 48. Narce M, Bellenger J, Rialland M, Bellenger S (2012) Recent advances on
stearoyl-Coa desaturase regulation in fatty liver diseases. Curr Drug Metab. 13:
1454–1463. 38. Zhu X, Topouzis S, Liang LF, Stotish RL (2004) Myostatin signaling through
Smad2, Smad3 and Smad4 is regulated by the inhibitory Smad7 by a negative
feedback mechanism. Cytokine. 26: 262–272. 49. Yao Z (2012) Human apolipoprotein C-III - a new intrahepatic protein factor
promoting assembly and secretion of very low density lipoproteins. Cardiovasc
Hematol Disord Drug Targets. 12: 133–140. 39. Kardassis D, Pardali K, Zannis VI (2000) SMAD proteins transactivate the
human ApoCIII promoter by interacting physically and functionally with
hepatocyte nuclear factor 4. J Biol Chem. 275: 41405–41414. 50. Buday A, Orsy P, Godo M, Mozes M, Kokeny G, et al. (2010) Elevated systemic
TGF-beta impairs aortic vasomotor function through activation of NADPH
oxidase-driven superoxide production and leads to hypertension, myocardial
remodeling, and increased plaque formation in apoE(2/2) mice. Am J Physiol
Heart Circ Physiol. 299: H386–395. p
y
40. Dong C, Zhu S, Wang T, Yoon W, Goldschmidt-Clermont PJ (2002)
Upregulation of PAI-1 is mediated through TGF-beta/Smad pathway in
transplant arteriopathy. J Heart Lung Transplant. 21: 999–1008. 41. Brezniceanu ML, Wei CC, Zhang SL, Hsieh TJ, Guo DF, et al. (2006)
Transforming growth factor-beta 1 stimulates angiotensinogen gene expression
in kidney proximal tubular cells. Kidney Int. 69: 1977–1985. 51. References Zincarelli C, Soltys S, Rengo G, Rabinowitz JE (2008) Analysis of AAV
serotypes 1–9 mediated gene expression and tropism in mice after systemic
injection. Mol Ther. 16: 1073–1080. 31. Lee SJ, McPherron AC (2001) Regulation of myostatin activity and muscle
growth. Proc Natl Acad Sci USA. 98: 9306–9311. 14. Bostick B, Ghosh A, Yue Y, Long C, Duan D (2007) Systemic AAV-9
transduction in mice is influenced by animal age but not by the route of
administration. Gene Ther. 14: 1605–1609. 32. Lee SJ, Reed LA, Davies MV, Girgenrath S, Goad ME, et al. (2005) Regulation
of muscle growth by multiple ligands signaling through activin type II receptors. Proc Natl Acad Sci USA. 102: 18117–18122. 15. Zang M, Zuccollo A, Hou X, Nagata D, Walsh K, et al (2004) AMP-activated
protein kinase is required for the lipid-lowering effect of metformin in insulin-
resistant human HepG2 cells. J Biol Chem. 279: 47898–47905. 33. Feijen A, Goumans MJ, van den Eijnden-van Raaij AJ (1994) Expression of
activin subunits, activin receptors and follistatin in postimplantation mouse
embryos suggests specific developmental functions for different activins. Development. 120: 3621–3637. 16. Clavreul N, Adachi T, Pimental DR, Ido Y, Schoneich C, et al. (2006) S-
glutathiolation by peroxynitrite of p21ras at cysteine-118 mediates its direct
activation and downstream signaling in endothelial cells. FASEB J. 20: 518–520. 34. Abou-Shady M, Baer HU, Friess H, Berberat P, Zimmermann A, et al. (1999)
Transforming growth factor betas and their signaling receptors in human
hepatocellular carcinoma. Am J Surg. 177: 209–215. 17. Kaikaus RM, Sui Z, Lysenko N, Wu NY, Ortiz de Montellano PR, et al. (1993)
Regulation of pathways of extramitochondrial fatty acid oxidation and liver fatty
acid-binding protein by long-chain monocarboxylic fatty acids in hepatocytes. Effect of inhibition of carnitine palmitoyltransferase I. J Biol Chem. 268: 26866–
26871. 35. Chen W, Woodruff TK, Mayo KE (2000) Activin A-induced HepG2 liver cell
apoptosis: involvement of activin receptors and smad proteins. Endocrinology. 141: 1263–1272. August 2013 | Volume 8 | Issue 8 | e71017 August 2013 | Volume 8 | Issue 8 | e71017 9 PLOS ONE | www.plosone.org Myostatin, Hepatosteatosis, and Atherosclerosis Hittel DS, Berggren JR, Shearer J, Boyle K, Houmard JA (2009) Increased
secretion and expression of myostatin in skeletal muscle from extremely obese
women. Diabetes. 58: 30–38. 42. Samuel W, Nagineni CN, Kutty RK, Parks WT, Gordon JS, et al. (2002)
Transforming growth factor-beta regulates stearoyl coenzyme A desaturase
expression through a Smad signaling pathway. J Biol Chem. 277: 59–66. 52. Guernec A, Chevalier B, Duclos MJ (2004) Nutrient supply enhances both IGF-I
and MSTN mRNA levels in chicken skeletal muscle. Domest Anim Endocrinol. 26: 143–154. 43. Ehnert S, Knobeloch D, Blankenstein A, Muller A, Bocker U, et al. (2010)
Neohepatocytes from alcoholics and controls express hepatocyte markers and
display reduced fibrogenic TGF-ss/Smad3 signaling: advantage for cell
transplantation? Alcohol Clin Exp Res. 34: 708–718. 53. Milan G, Dalla Nora E, Pilon C, Pagano C, Granzotto M, et al. (2004) Changes
in muscle myostatin expression in obese subjects after weight loss. J Clin
Endocrinol Metab. 89: 2724–2727. 54. Allen DL, Cleary AS, Speaker KJ, Lindsay SF, Uyenishi J, et al (2008). Myostatin, activin receptor IIb, and follistatin-like-3 gene expression are altered
in adipose tissue and skeletal muscle of obese mice. Am J Physiol Endocrinol
Metab. 294: E918–927. 44. Rebbapragada A, Benchabane H, Wrana JL, Celeste AJ, Attisano L (2003)
Myostatin signals through a transforming growth factor beta-like signaling
pathway to block adipogenesis. Mol Cell Biol. 23: 7230–7242. 45. Ponnuswamy P, Schrottle A, Ostermeier E, Gruner S, Huang PL, et al. (2012)
eNOS protects from atherosclerosis despite relevant superoxide production by
the enzyme in apoE mice. PLoS One. 7: e30193. 55. Lyons JA, Haring JS, Biga PR (2010) Myostatin expression, lymphocyte
population, and potential cytokine production correlate with predisposition to
high-fat diet induced obesity in mice. PLoS One. 5: e12928. y
p
46. Jung J, Ko SH, Yoo do Y, Lee JY, Kim YJ, et al. (2012) 5,7-Dihydroxy-3,4,6-
trimethoxyflavone inhibits intercellular adhesion molecule 1 and vascular cell PLOS ONE | www.plosone.org August 2013 | Volume 8 | Issue 8 | e71017 PLOS ONE | www.plosone.org 10
|
https://openalex.org/W4282015721
|
https://cyberleninka.ru/article/n/opredelenie-stranovyh-osobennostey-tsifrovizatsii-v-gosudarstvah-eaes/pdf
|
Russian
| null |
Определение страновых особенностей цифровизации в государствах ЕАЭС
|
Trudy BGTU. Seriâ 5. Èkonomika i upravlenie
| 2,022
|
cc-by
| 5,277
|
УДК 502.17 И. В. Новикова, А. В. Равино
Белорусский государственный технологический университет ОПРЕДЕЛЕНИЕ СТРАНОВЫХ ОСОБЕННОСТЕЙ ЦИФРОВИЗАЦИИ
В ГОСУДАРСТВАХ ЕАЭС Òðóäû ÁÃÒÓ Ñåðèÿ 5 № 1 2022 6 Îïðåäåëåíèå ñòðàíîâûõ îñîáåííîñòåé öèôðîâèçàöèè â ãîñóäàðñòâàõ ÅÀÝÑ The purpose of the study is to determine the features of digitalization in the EAEU member states
according to international ratings of digital development. The article contains an analysis of the place of
the EAEU member states in digital rankings based on: ICT Development Index, Networked Readiness
Index, Digital Adoption Index, UN Global E-Government Development Index, E-Participation Index,
GovTech Maturity Index, Global Cybersecurity Index. An assessment of the position of Belarus in seven
digital ratings is given. The features of digitalization of the EAEU member countries are studied. This is
important for further research work in order to assess the threats and opportunities of digitalization in the
EAEU and develop mechanisms for the institutional environment of the digital economy. Key words: digitalization, rating, index, international ratings of digital development, Eurasian
Economic Union, Belarus. For citation: Novikova I. V., Ravino A. V. Determining the features of digitalization of the EAEU
member states. Proceedings of BSTU, issue 5, Economics and Management, 2022, no. 1 (256), pp. 5–12
(In Russian). For citation: Novikova I. V., Ravino A. V. Determining the features of digitalization of the EAEU
member states. Proceedings of BSTU, issue 5, Economics and Management, 2022, no. 1 (256), pp. 5–12
(In Russian). Для реализации региональной цифровой
политики, использования инструментов циф-
рового взаимодействия и формирования меха-
низмов надгосударственного регулирования
целесообразным является сравнительный ана-
лиз уровня развития цифровизации во всех го-
сударствах – членах ЕАЭС. Целью исследова-
ния выступает определение страновых особен-
ностей цифровизации в ЕАЭС и анализ позиции
Беларуси в сравнении со странами – членами
Союза. Введение. Цифровизация, являясь по своему
характеру эндогенным, технологическим факто-
ром экономического развития, становится важ-
ным его институтом в современных условиях. Институционализация цифровой экономики –
методологически сложный процесс, с необходи-
мостью требующий учета временного фактора и
последовательности действий государственного
и частного секторов с целью придания непроти-
воречивого характера и динамики экономиче-
ским процессам [1, 2]. Введение. Цифровизация, являясь по своему
характеру эндогенным, технологическим факто-
ром экономического развития, становится важ-
ным его институтом в современных условиях. Институционализация цифровой экономики –
методологически сложный процесс, с необходи-
мостью требующий учета временного фактора и
последовательности действий государственного
и частного секторов с целью придания непроти-
воречивого характера и динамики экономиче-
ским процессам [1, 2]. ОПРЕДЕЛЕНИЕ СТРАНОВЫХ ОСОБЕННОСТЕЙ ЦИФРОВИЗАЦИИ
В ГОСУДАРСТВАХ ЕАЭС Цифровизация – это процесс проникновения цифровых технологий во все аспекты человече-
ской деятельности. Для Беларуси цифровизация выступает ведущим направлением развития. В условиях глобализации цифровая трансформация должна осуществляться не только в нацио-
нальной экономической системе, но и в рамках интеграционных группировок. Беларусь является
членом Евразийского экономического союза (ЕАЭС). Евразийская интеграция выступает важным
фактором цифровизации не только экономики Беларуси, но стран – членов Союза. Для реализации
региональной цифровой политики, использования механизмов цифрового взаимодействия стран
и формирования единого надгосударственного регулирования целесообразным является сравни-
тельный анализ уровня цифровизации во всех государствах ЕАЭС. Целью исследования выступает определение страновых особенностей цифровизации в стра-
нах – членах ЕАЭС по международным рейтингам цифрового развития. В статье проведен анализ
положения стран ЕАЭС в цифровых рейтингах на основе: индекса развития ИКТ, индекса сетевой
готовности, индекса цифрового внедрения, индекса развития электронного правительства, ин-
декса электронного участия, индекса цифровой зрелости в категории «GovTech», глобального ин-
декса кибербезопасности. Дана оценка положения Беларуси в семи цифровых рейтингах. Выявлены страновые особенности цифровизации стран – членов ЕАЭС, которые будут
учтены при проведении дальнейшей научно-исследовательской работы для прогнозирования
угроз и возможностей развития цифровой экономики на уровне интеграционной группировки
ЕАЭС и разработки инфраструктурных механизмов, обеспечивающих создание адекватной ин-
ституциональной среды для развития цифровой экономики. Ключевые слова: цифровизация, рейтинг, индекс, международные рейтинги цифрового раз
вития, Евразийский экономический союз, Беларусь. Для цитирования: Новикова И. В., Равино А. В. Определение страновых особенностей циф-
ровизации в государствах ЕАЭС // Труды БГТУ. Сер. 5, Экономика и управление. 2022. № 1 (256). С. 5–12. I. V. Novikova, А. V. Ravino
Belarusian State Technological University
DETERMINING THE FEATURES OF DIGITALIZATION
OF THE EAEU MEMBER STATES I. V. Novikova, А. V. Ravino
Belarusian State Technological University
DETERMINING THE FEATURES OF DIGITALIZATION
OF THE EAEU MEMBER STATES Digitalization is the process of penetration of digital technologies into all aspects of human activity. Digitalization is the main direction of development of Belarus. With globalization, digitalization develops
not only in the economy of one country, but also in integration unions. Belarus is a member of the
Eurasian Economic Union (EAEU). Eurasian integration is an important factor in the digitalization of the
economy of Belarus and the EAEU member states. A comparative analysis of digitalization in the EAEU
states is necessary for regional digital policy, digital interaction between states and the formation of a
unified system of supranational regulation. ОПРЕДЕЛЕНИЕ СТРАНОВЫХ ОСОБЕННОСТЕЙ ЦИФРОВИЗАЦИИ
В ГОСУДАРСТВАХ ЕАЭС В условиях глобализации мировой эконо-
мики формирование институциональной среды и
инструментария регулирования цифрового раз-
вития в Беларуси должно проходить не только в
национальной экономической системе, но и в
рамках интеграционных группировок. Беларусь
входит в число активных участников интеграци-
онных процессов. По данным Министерства ино-
странных дел Республики Беларусь, наша страна
является членом (или наблюдателем) около 60 ин-
теграционных объединений, международных и
региональных организаций, специализированных
учреждений ООН. Основная часть. Последнее десятилетие ха-
рактеризуется ростом оценочных исследований
цифровой экономики, ее отдельных сфер и ин-
фраструктурных составляющих международ-
ных организаций, итогом которых выступают
мировые рейтинги. Методология оценки уровня
цифровизации в рейтинге базируется на расчете
индексов, включающих субиндексы. Субин-
дексы показывают уровень развития страны по
одному из направлений цифровой экономики. Информационной основой рейтингов являются
статистические данные, базы данных междуна-
родных организаций, результаты опросов экс-
пертов и общественности [4]. у р
Беларусь входит в состав Евразийского эко-
номического союза (ЕАЭС). Евразийская инте-
грация выступает не только важным фактором
успешного развития экономики Беларуси и
укрепления ее позиций в мировом хозяйстве,
но и фактором динамичного развития стран –
членов Союза. Следовательно, от активизации
регионального взаимодействия зависит и циф-
ровизация как экономики Республики Бела-
русь, так и евразийской экономики. Эксперты
Всемирного банка прогнозируют, что экономи-
ческий эффект от реализации единой цифровой
политики ЕАЭС к 2025 г. увеличит ВВП Союза
минимум на 10,6% от общего ожидаемого ро-
ста совокупного ВВП государств-членов, что в
2 раза превышает возможный размер роста
ВВП стран ЕАЭС без общей цифровой по-
вестки [3]. Оценка страны, ее положения в мировых
рейтингах оказывает влияние на политические и
экономические решения. Для многих стран по-
зиции в международных рейтингах стали целе-
выми показателями в национальной системе
стратегического планирования. Анализ положения стран ЕАЭС в цифровых
рейтингах. Проведем сравнительный анализ
уровня цифровизации в странах ЕАЭС по следу-
ющим международным экспертным рейтингам,
отражающим цифровое развитие [1, 4–17]: – рейтинг по уровню развития информаци-
онно-коммуникационных технологий (ИКТ) на
основе расчета индекса развития ИКТ (ICT
Development Index – IDI). IDI призван сравни-
вать достижения стран в освоении ИКТ. IDI
включает 11 показателей, агрегируемых в три Òðóäû ÁÃÒÓ Ñåðèÿ 5 № 1 2022 È. Â. Íîâèêîâà, À. Â. Ðàâèíî 7 субиндекса, которые оценивают доступ, исполь-
зование и знания ИКТ [5, 6]; граждан, механизмы внедрения решений «Gov-
Tech» [15]; – рейтинг по уровню кибербезопасности на ос-
нове расчета глобального индекса кибербезопас-
ности (Global Cybersecurity Index – GCI). GCI
призван сравнивать информационную безопас-
ность, а также управление критической инфра-
структурой Интернета стран мира. ОПРЕДЕЛЕНИЕ СТРАНОВЫХ ОСОБЕННОСТЕЙ ЦИФРОВИЗАЦИИ
В ГОСУДАРСТВАХ ЕАЭС GCI включает
17 показателей, агрегируемых в пять основных
категорий: правовые меры, технические меры,
организационные меры, создание потенциала, со-
трудничество [16, 17]. – рейтинг по уровню сетевой готовности на
основе расчета индекса сетевой готовности
(Networked Readiness Index – NRI). NRI отра-
жает потенциал стран мира для использования
возможностей ИКТ в целях цифровой трансфор-
мации. NRI включает 53 показателя, агрегируе-
мых в 10 микроиндексов и в 4 субиндекса, кото-
рые оценивают среду для развития ИКТ, готов-
ность общества к использованию, фактическое
использование и последствия (эффект) внедре-
ния ИКТ в экономике и обществе [7, 8]; В таблице представлены позиции государств –
членов ЕАЭС в семи международных рейтингах
цифрового развития. – рейтинг по уровню цифрового внедрения
на основе расчета индекса цифрового внедрения
(Digital Adoption Index – DAI). DAI призван ана-
лизировать достижения стран по распростране-
нию и использованию цифровых технологий. DAI включает 9 показателей, агрегируемых в
три субиндекса: бизнес (цифровое внедрение в
сфере бизнеса), население (цифровой доступ для
граждан), государство (цифровизация госсек-
тора страны) [9, 10]; – рейтинг по уровню цифрового внедрения
на основе расчета индекса цифрового внедрения
(Digital Adoption Index – DAI). DAI призван ана-
лизировать достижения стран по распростране-
нию и использованию цифровых технологий. DAI включает 9 показателей, агрегируемых в
три субиндекса: бизнес (цифровое внедрение в
сфере бизнеса), население (цифровой доступ для
граждан), государство (цифровизация госсек-
тора страны) [9, 10]; Анализ позволил выявить следующие особен-
ности цифровизации государств – членов ЕАЭС. Среди представленных рейтингов, характеризу-
ющих процесс цифровизации, в блоке стран
ЕАЭС Россия является лидером (среднее значе-
ние места в рассмотренных рейтингах – 32). В четырех рейтингах из семи Россия занимает ве-
дущее положение среди стран – членов ЕАЭС: рей-
тинг стран мира по уровню сетевой готовности,
рейтинг по уровню цифрового внедрения, рейтинг
по уровню цифровой зрелости в категории «Gov-
Tech», рейтинг стран по уровню кибербезопасно-
сти. В двух из анализируемых рейтингов лидером
является Казахстан: рейтинг стран мира по уровню
развития электронного правительства и рейтинг по
уровню цифрового участия. В рейтинге стран мира
по уровню развития ИКТ лидером является Бела-
русь. Худшее положение из государств ЕАЭС по
показателям рейтингов цифрового развития у Ар-
мении и Кыргызстана (средние значения места в
рассмотренных рейтингах 68 и 90 соответственно). – рейтинг по уровню развития электронного
правительства на основе расчета индекса разви-
тия электронного правительства (UN Global
E-Government Development Index – EGDI). EGDI
оценивает уровень использования ИКТ в стране
для предоставления гражданам государствен-
ных услуг. ОПРЕДЕЛЕНИЕ СТРАНОВЫХ ОСОБЕННОСТЕЙ ЦИФРОВИЗАЦИИ
В ГОСУДАРСТВАХ ЕАЭС Кыргызстан, Армения и Беларусь
отстают по следующим позициям: государства – участники ЕАЭС разделяются на
две группы: группа «зарождающихся» (Кыргыз-
стан) и группа «переходных» (Казахстан, Бела-
русь, Россия и Армения) цифровых экономик [18]. Кроме того, в силу неоднородного инфраструк-
турного и экономического развития внутри стран
разрыв присутствует и на локальном, местном
уровне: наблюдается значительная разница в уров-
не цифровизации городской и сельской среды. у
– обучение навыкам кибербезопасности, адап-
тированным к потребностям малых и средних
предприятий; – усиление кибербезопасности в финансовом
секторе, здравоохранении, энергетике и других
ключевых секторах национальной экономики; – защита критической инфраструктуры, ко-
торую необходимо совершенствовать с целью
борьбы с возникающими киберугрозами; На основе проведенного анализа страновых
особенностей цифрового развития государств
ЕАЭС можно сделать вывод, что ЕАЭС –
это устойчивое образование, которое показало
свою целесообразность и эффективность, страны
Союза располагаются на достойном уровне в
цифровых рейтингах, но имеются проблемные
места, которые требуют подробного анализа и
выявления причин, мешающих цифровой транс-
формации. В вопросах формирования и развития
цифровой экономики рекомендации для Союза
сохраняются, как и по другим общим направле-
ниям развития, а именно: страны с высоким уров-
нем цифровых возможностей должны оказывать
поддержку другим менее развитым в данном на-
правлении странам – членам группировки в целях
активизации процесса развития цифрового Союза. 4) в экономических моделях стран – участ-
ниц Союза, в которых относительно медлен-
ными темпами происходит внедрение цифровых
технологий, поэтому необходимы меры по со-
зданию и модернизации коммуникационных ин-
фраструктур для удовлетворения растущего
спроса на услуги ИКТ; 5) в существующем цифровом разрыве меж-
ду странами – членами ЕАЭС, асимметричных
возможностях и рисках цифровой экономики
для различных стран Союза, которые высту-
пают сдерживающими факторами внедрения
прорывных технологий и цифровой трансфор-
мации. По классификации Всемирного банка Глобальные баллы и позиции стран ЕАЭС в международных рейтингах,
характеризующих цифровое развитие
Рейтинг
Индекс
Организатор
Количество стран
в рейтинге
Год
Страны (место/индекс)
Лучшее значение
(место/индекс)
Худшее значение
(место/индекс)
Среднее значение
по ЕАЭС (место/индекс)
Среднемировое
значение индекса
Армения
Беларусь
Казахстан
Кыргызстан
Россия
1. Рейтинг стран по уров-
ню развития ИКТ
IDI
ITU
176 2017
75
32
52
109
45
32
109
63
5,11
5,76
7,55
6,79
4,37
7,07
7,55
4,37
6,31
2. Рейтинг стран по уров-
ню сетевой готовности
NRI WEF, WB,
INSEAD 134 2020
55
65
56
94
48
48
94
64
49,90
51,91 49,16 51,38 38,60 54,23 54,23 38,60 47,80
3. ОПРЕДЕЛЕНИЕ СТРАНОВЫХ ОСОБЕННОСТЕЙ ЦИФРОВИЗАЦИИ
В ГОСУДАРСТВАХ ЕАЭС EGDI включает показатели, агреги-
руемые в три субиндекса: возможность и каче-
ство онлайн-сервисов (среда для развития ИКТ),
развитость телекоммуникационной инфраструк-
туры (фактическое использование ИКТ), чело-
веческий капитал (уровень образованности
населения [11–13]; Если сравнивать среднее значение индексов в
рейтингах по ЕАЭС со среднемировым значением,
то интегрированный показатель по ЕАЭС в IDI,
DAI, EGDI, EPART, GCI выше среднемирового. – рейтинг по уровню цифрового участия на
основе расчета индекса электронного участия
(E-Participation Index – EPART). EPART допол-
няет EGDI и служит для оценки уровня развития
сервисов цифровой коммуникации между граж-
данами и государством. EPART включает три
субиндекса: Е-информация (электронное ин-
формирование граждан правительством), Е-кон-
сультации (привлечение граждан к обсуждению
вопросов государственной политики), Е-реше-
ния (принятие решений на основе электронных
технологий) [14]; Вместе с тем в странах ЕАЭС сохраняются сле-
дующие негативные особенности цифровизации: 1) во внедрении решений «GovTech», т. е. циф-
ровых технологий в государственном секторе. Страны – члены ЕАЭС уступают странам – лиде-
рам в области «GovTech», которые используют
передовые цифровые решения и генерируют ми-
ровой опыт на всех четырех направлениях
«GovTech»: деятельность правительства, оказа-
ние государственных услуг, вовлеченность граж-
дан, механизмы внедрения решений «GovTech»; – рейтинг по уровню цифровой зрелости в
категории «GovTech» на основе расчета индекса
цифровой зрелости в категории «GovTech»
(GovTech Maturity Index – GTMI). Индекс оце-
нивает уровень внедрения решений «Govern-
ment Technologies» («GovTech») в государствен-
ный сектор страны. GTMI включает 49 показате-
лей, агрегируемых в четыре компонента: основ-
ные направления деятельности правительства,
оказание государственных услуг, вовлеченность – рейтинг по уровню цифровой зрелости в
категории «GovTech» на основе расчета индекса
цифровой зрелости в категории «GovTech»
(GovTech Maturity Index – GTMI). Индекс оце-
нивает уровень внедрения решений «Govern-
ment Technologies» («GovTech») в государствен-
ный сектор страны. GTMI включает 49 показате-
лей, агрегируемых в четыре компонента: основ-
ные направления деятельности правительства,
оказание государственных услуг, вовлеченность 2) в логистической инфраструктуре форми-
рования эффективной электронной коммерции
(e-commerce). Организованная логистическая ин-
фраструктура может значительно усилить эффект
от электронной коммерции, способствуя малым и
средним предприятиям стран – членов ЕАЭС эф-
фективно осуществлять торговую деятельность; 2) в логистической инфраструктуре форми-
рования эффективной электронной коммерции
(e-commerce). Организованная логистическая ин-
фраструктура может значительно усилить эффект
от электронной коммерции, способствуя малым и
средним предприятиям стран – членов ЕАЭС эф-
фективно осуществлять торговую деятельность; Òðóäû ÁÃÒÓ Ñåðèÿ 5 № 1 2022 Òðóäû ÁÃÒÓ Ñåðèÿ 5 № 1 2022 Îïðåäåëåíèå ñòðàíîâûõ îñîáåííîñòåé öèôðîâèçàöèè â ãîñóäàðñòâàõ ÅÀÝÑ 8 8 3) в создании потенциала в области кибер-
безопасности. Примечание. B – страны, уделяющие значительное внимание решениям «GovTech». C – страны, уделяющие опреде-
ленное внимание решениям «GovTech». ание. B – страны, уделяющие значительное внимание решениям «GovTech». C – страны, уделяющие опреде
мание решениям «GovTech». ОПРЕДЕЛЕНИЕ СТРАНОВЫХ ОСОБЕННОСТЕЙ ЦИФРОВИЗАЦИИ
В ГОСУДАРСТВАХ ЕАЭС Рейтинг стран по уров-
ню цифрового внедрения DAI
WB
180 2016
60
72
45
96
28
28
96
60
0,516
0,62
0,59
0,67 0,499
0,74
0,74 0,499 0,62
4. Рейтинг стран по уров-
ню развития электронного
правительства
EGDI UNDESA 193 2020
68
40
29
83
36
29
83
51
0,5988
0,7136 0,8084 0,8375 0,6749 0,8244 0,8375 0,6749 0,770
5. Рейтинг стран по уров-
ню цифрового участия
EPART UNDESA 193 2020
57
57
26
66
27
26
66
47
0,57
0,750 0,750 0,881 0,7143 0,869 0,881 0,7143 0,793
6. Рейтинг стран по уров-
ню цифровой зрелости
GTMI
WB
198 2020
B
C
B
B
B
B
C
B–C
–
7. Рейтинг стран по уров-
ню кибербезопасности
GCI
ITU
194 2020
90
89
31
92
5
5
92
61
52,50
50,47 50,57 93,15 49,64 98,06 98,06 49,64 68,40
Лучшее место в рейтинге по стране – члену ЕАЭС
55
32
26
66
5
–
–
–
–
Среднее значение места в рейтинге по стране – члену ЕАЭС
68
59
40
90
32
–
–
–
– Глобальные баллы и позиции стран ЕАЭС в международных рейтингах,
характеризующих цифровое развитие Òðóäû ÁÃÒÓ Ñåðèÿ 5 № 1 2022 È. Â. Íîâèêîâà, À. Â. Ðàâèíî 9 Для дальнейшего развития цифровизации
следует имплементировать опыт зарубежных
стран и интеграционных образований – лидеров
цифровых рейтингов; развивать национальный
и интеграционный цифровой контент госу-
дарств ЕАЭС; активно использовать потенциал
и потребности партнеров ЕАЭС, а также иных
региональных и международных объединений;
увеличивать экспорт-импорт технологий на ос-
нове цифровых возможностей и механизмов,
предоставленных Союзом. позволяющие оказывать населению, бизнесу и
правительственному сектору экономики Бела-
руси качественные услуги фиксированного и
мобильного широкополосного доступа в Интер-
нет (IDI-2017, EGDI-2020); – отмечается высокое качество националь-
ной образовательной системы в сфере ИКТ от
школьного, среднего профессионального, выс-
шего до переподготовки взрослого населения
(IDI-2017); – цифровое обеспечение бизнеса (количе-
ство бизнес-организаций с веб-сайтами, защи-
щенных серверов; скорость загрузки и пр.) в рес-
публике оценивается высоко (DAI-2016) при не-
достаточном уровне цифрового взаимодействия
«государство – бизнес» (GTMI-2020); Анализ положения Республики Беларусь в
цифровых рейтингах. Оценка уровня цифрови-
зации Беларуси по показателям международных
экспертных рейтингов показала [1, 4–17]: – в семи представленных рейтингах, харак-
теризующих процесс цифровизации и проник-
новения ИКТ, Республика Беларусь (по данным
последних
рейтингов: DAI-2016, IDI-2017,
NRI-2020, EGDI-2020, EPART-2020, GTMI-2020,
GCI-2020) в среднем занимает 59 место среди
стран мира (46 место с учетом количества стран,
задействованных в рейтингах: от 134 до 198 стран
в зависимости от рейтинга). ОПРЕДЕЛЕНИЕ СТРАНОВЫХ ОСОБЕННОСТЕЙ ЦИФРОВИЗАЦИИ
В ГОСУДАРСТВАХ ЕАЭС Лучшие позиции
Беларусь заняла в рейтинге стран мира по
уровню развития ИКТ, рейтинге по уровню циф-
рового участия и рейтинге стран мира по
уровню развития электронного правительства
(рисунок). Динамика мест республики в рейтин-
гах нестабильна. По отдельным индексам пони-
жение места страны происходило не за счет
ослабления позиции Беларуси по оцениваемым
параметрам, а из-за усиления позиций других
государств (IDI, EGDI); – цифровая защита Беларуси требует даль-
нейшего развития, и если международным ини-
циативам и межгосударственному сотрудниче-
ству в сфере кибербезопасности дана достаточно
высокая оценка в рейтингах, то организационные
меры по информационной компьютерной без-
опасности (стратегии, использование нацио-
нальных инструментов сравнительного анализа)
и развитие потенциала цифровой безопасности
(внедрение стандартизации и сертификации)
нуждаются в корректировке (GCI-2020); – существенное влияние на показатели рей-
тингов цифрового развития Беларуси за 2021 г. и дальнейший период будут оказывать меры
адаптации национальной экономики к усло-
виям пандемии COVID-19, условиям внедре-
ния цифровых технологий практически во все
сферы деятельности: от бизнеса, перехода на
удаленную работу, здравоохранения, образова-
ния до быта. – в республике создана развитая ИТК-ин-
фраструктура и доступный цифровой контент, Беларусь в международных рейтингах, характеризующих цифровое развитие
(по данным последних рейтингов) [5–17]
89
57
40
72
65
32
194
193
193
180
134
176
0
50
100
150
200
250
Рейтинг стран по уровню кибербезопасности
Рейтинг по уровню цифрового участия
Рейтинг стран мира по уровню развития
электронного правительства
Рейтинг по уровню цифрового внедрения
Рейтинг стран мира по уровню сетевой готовности
Рейтинг стран мира по уровню развития ИКТ
Количество стран рейтинга
Место Республики Беларусь Беларусь в международных рейтингах, характеризующих цифровое развитие
(по данным последних рейтингов) [5–17] Òðóäû ÁÃÒÓ Ñåðèÿ 5 № 1 2022 Òðóäû ÁÃÒÓ Ñåðèÿ 5 № 1 2022 Îïðåäåëåíèå ñòðàíîâûõ îñîáåííîñòåé öèôðîâèçàöèè â ãîñóäàðñòâàõ ÅÀÝÑ 10 В целом Беларусь, по оценке международ-
ных экспертных организаций, относится к
группе стран, имеющих высокий цифровой по-
тенциал и демонстрирующих динамику разви-
тия ИКТ. Проводимая поступательная цифровая
политика Республики Беларусь позволит улуч-
шить позицию страны в международных рей-
тингах цифрового развития, в том числе за счет
дальнейшего развертывания сетей LTE в регио-
нах, развития спутниковой связи, облачных тех-
нологий, разработки и внедрения новых цифро-
вых сервисов и продуктов, передовых цифровых
технологий. ментов как государственного, так и надгосудар-
ственного регулирования с учетом потребностей и
возможностей партнеров по региональной груп-
пировке ЕАЭС с целью дальнейшего цифрового
развития и нивелировки возникающих угроз эко-
номике, обществу, гражданам. ОПРЕДЕЛЕНИЕ СТРАНОВЫХ ОСОБЕННОСТЕЙ ЦИФРОВИЗАЦИИ
В ГОСУДАРСТВАХ ЕАЭС Выявленные нами страновые особенности
цифровизации стран – членов ЕАЭС будут учтены
при проведении дальнейшей научно-исследова-
тельской работы «Разработать институциональ-
ные механизмы и инструментарий государствен-
ного регулирования для становления и развития
цифровой экономики, обеспечивающие нацио-
нальную безопасность и создающие условия для
развития интеграционных процессов в ЕАЭС» в
рамках ГПНИ «Общество и гуманитарная безопас-
ность белорусского государства», подпрограмма
«Экономика» в 2022–2025 гг. (руководитель – док-
тор экономических наук, профессор Новикова И. В.),
а именно: для прогнозирования возникающих и
потенциальных угроз при становлении и развитии
цифровой экономики на уровне интеграционной
группировки ЕАЭС; для оценки эффективности
наднационального регулирования, становления и
развития цифровой экономики в интеграционных
группировках; при разработке инфраструктурных
механизмов и инструментов, обеспечивающих со-
здание адекватной институциональной среды для
развития цифровой экономики при предотвраще-
нии угроз в национальной экономике и в интегра-
ционных процессах. Заключение. Перед странами – членами
ЕАЭС поставлена задача формирования единого
цифрового пространства в рамках цифровой по-
вестки ЕАЭС до 2025 г. Реализация общей циф-
ровой повестки способна обеспечить для госу-
дарств ЕАЭС: рост занятости в отрасли ИКТ;
прирост объема экспорта услуг ИКТ более чем
на 70%; развитие промышленной, производ-
ственной и научно-технической кооперации;
увеличение «цифрового» ВВП [18]. Анализ страновых особенностей цифровиза-
ции в ЕАЭС по показателям международных экс-
пертных рейтингов выявил, что страны евразий-
ского интеграционного проекта имеют значитель-
ные ресурсы для создания цифровой экономики. Сформировать устойчивую наднациональную
цифровую экосистему возможно при внедрении
новых институциональных механизмов и инстру- Список литературы 1. Разработать институциональные механизмы и инструментарий государственного регулирова-
ния для становления и развития цифровой экономики, обеспечивающие национальную безопасность
и создающие условия для развития интеграционных процессов в ЕАЭС: отчет о НИР (промеж.) / Бе-
лорус. гос. технол. ун-т (БГТУ); рук. И. В. Новикова. Минск, 2021. 211 с. № ГР 20211617. 1. Разработать институциональные механизмы и инструментарий государственного регулирова-
ния для становления и развития цифровой экономики, обеспечивающие национальную безопасность
и создающие условия для развития интеграционных процессов в ЕАЭС: отчет о НИР (промеж.) / Бе-
лорус. гос. технол. ун-т (БГТУ); рук. И. В. Новикова. Минск, 2021. 211 с. № ГР 20211617. ру
у
(
) ру
2. Новикова И. В. Цифровая техноэкономическая парадигма в смене стратегии цифровизации
Республики Беларусь // Труды БГТУ. Сер. 5, Экономика и управление. 2020. № 1 (232). С. 5–12. 2. Новикова И. В. Цифровая техноэкономическая парадигма в смене стратегии цифровизации
Республики Беларусь // Труды БГТУ. Сер. 5, Экономика и управление. 2020. № 1 (232). С. 5–12. 3. Цифровая
перезагрузка
ЕАЭС. Новый
взгляд
на
экономику
/
ЕЭК. URL:
http://www.eurasiancommission.org/ru/act/dmi/workgroup/Pages/Цифровая-перезагрузка-ЕАЭС--
Новый-взгляд-на-экономику%21.aspx (дата обращения: 30.01.2022). 3. Цифровая
перезагрузка
ЕАЭС. Новый
взгляд
на
экономику
/
ЕЭК. URL:
http://www.eurasiancommission.org/ru/act/dmi/workgroup/Pages/Цифровая-перезагрузка-ЕАЭС--
Новый-взгляд-на-экономику%21.aspx (дата обращения: 30.01.2022). д
у
p
(д
р щ
)
4. Головенчик Г. Г. Цифровизация белорусской экономики в современных условиях глобализа-
ции. Минск: Издат. центр БГУ, 2019. 257 с. у
p
(
р
)
4. Головенчик Г. Г. Цифровизация белорусской экономики в современных условиях глобализа-
ции. Минск: Издат. центр БГУ, 2019. 257 с. 5. Рейтинг стран мира по уровню развития информационно-коммуникационных технологий. URL: http://gtmarket.ru/ratings/ict-development-index (дата обращения: 30.01.2022). 5. Рейтинг стран мира по уровню развития информационно-коммуникационных технологий. URL: http://gtmarket.ru/ratings/ict-development-index (дата обращения: 30.01.2022). p
g
g
p
(
р
)
6. International Telecommunication Union (ITU). URL: http://www.itu.int/ (date of access: 30.01.2022). 7. Рейтинг стран мира по индексу сетевой готовности. URL: http://gtmarket.ru/ratings/networked-
readiness-index (дата обращения: 30.01.2022). 6. International Telecommunication Union (ITU). URL: http://www.itu.int/ (date of access: 30.01.2022). 7. Рейтинг стран мира по индексу сетевой готовности. URL: http://gtmarket.ru/ratings/networked-
readiness-index (дата обращения: 30.01.2022). (
р
)
8. Network Readiness Index. URL: http://networkreadinessindex.org/ (date of access: 30.01.2022 (
р
)
8. Network Readiness Index. URL: http://networkreadinessindex.org/ (date of access: 30.01.2022). 9. Digital Adoption Index / World Bank. URL: http://www.worldbank.org/en/publication/wdr2016/
Digital-Adoption-Index (date of access: 30.01.2022). p
g (
)
9. Digital Adoption Index / World Bank. URL: http://www.worldbank.org/en/publication/wdr2016/
Digital-Adoption-Index (date of access: 30.01.2022). 10. Digital Adoption Index (DAI): Measuring the Global Spread of Digital Technologies. URL:
http://thedocs.worldbank.org/en/doc/587221475074960682-0050022016/original/WDR16BPDAImethodology.pdf
(date of access: 30.01.2022). References 1. Razrabotat’ institutsionalʼnyye mekhanizmy i instrumentariy gosudarstvennogo regulirovaniya dlya
stanovleniya i razvitiya tsifrovoy ekonomiki, obespechivayushchiye natsionalʼnuyu bezopasnostʼ i
sozdayushchiye usloviya dlya razvitiya integratsionnykh protsessov v EAES [Develop institutional
arrangements and tools state regulation for the development of the digital economy, ensuring national
security and the development of integration processes in the EAEU]. Head I. V. Novikova. Minsk, 2021. 211 p. No. GR 20211617 (In Russian). 1. Razrabotat’ institutsionalʼnyye mekhanizmy i instrumentariy gosudarstvennogo regulirovaniya dlya
stanovleniya i razvitiya tsifrovoy ekonomiki, obespechivayushchiye natsionalʼnuyu bezopasnostʼ i
sozdayushchiye usloviya dlya razvitiya integratsionnykh protsessov v EAES [Develop institutional
arrangements and tools state regulation for the development of the digital economy, ensuring national
security and the development of integration processes in the EAEU]. Head I. V. Novikova. Minsk, 2021. 211 p. No. GR 20211617 (In Russian). 1. Razrabotat’ institutsionalʼnyye mekhanizmy i instrumentariy gosudarstvennogo regulirovaniya dlya
stanovleniya i razvitiya tsifrovoy ekonomiki, obespechivayushchiye natsionalʼnuyu bezopasnostʼ i
sozdayushchiye usloviya dlya razvitiya integratsionnykh protsessov v EAES [Develop institutional
arrangements and tools state regulation for the development of the digital economy, ensuring national
security and the development of integration processes in the EAEU]. Head I. V. Novikova. Minsk, 2021. 211 p. No. GR 20211617 (In Russian). p
(
)
2. Novikova I. V. Digital techno-economic paradigm in changing the digitalization strategy of the
Republic of Belarus. Trudy BGTU [Proceedings of BSTU], series 5, Economics and Management, 2020,
no. 1 (232), pp. 5–12 (In Russian). 2. Novikova I. V. Digital techno-economic paradigm in changing the digitalization strategy of the
Republic of Belarus. Trudy BGTU [Proceedings of BSTU], series 5, Economics and Management, 2020,
no. 1 (232), pp. 5–12 (In Russian). (
), pp
(
)
3. Digital reset of the EAEU. New look at the deal. Available at: http://www.eurasiancommission.org/
ru/act/dmi/workgroup/Pages/Цифровая-перезагрузка-ЕАЭС--Новый-взгляд-на-экономику%21.aspx (accessed
30.01.2022) (In Russian). (
), pp
(
)
3. Digital reset of the EAEU. New look at the deal. Available at: http://www.eurasiancommission.org/
ru/act/dmi/workgroup/Pages/Цифровая-перезагрузка-ЕАЭС--Новый-взгляд-на-экономику%21.aspx (accessed
30.01.2022) (In Russian). ) (
)
4. Golovenchik G. G. Tsifrovizatsiya belorusskoy ekonomiki v sovremennykh usloviyakh globalizatsii
[Digitalization of the Belarusian economy in the current conditions of globalization]. Minsk, Izdatelʼskiy
tsentr BGU Publ., 2019. 257 p. (In Russian). ) (
)
4. Golovenchik G. G. Tsifrovizatsiya belorusskoy ekonomiki v sovremennykh usloviyakh globalizatsii
[Digitalization of the Belarusian economy in the current conditions of globalization]. Minsk, Izdatelʼskiy
tsentr BGU Publ., 2019. 257 p. (In Russian). p (
)
5. Список литературы (
)
11. Рейтинг стран мира по индексу развития электронного правительства. URL: http://gtmarket.ru/
ratings/e-government-development-index (дата обращения: 30.01.2022). 11. Рейтинг стран мира по индексу развития электронного правительства. URL: http://gtmarket.ru/
ratings/e-government-development-index (дата обращения: 30.01.2022). Òðóäû ÁÃÒÓ Ñåðèÿ 5 № 1 2022 È. Â. Íîâèêîâà, À. Â. Ðàâèíî 11 12. United Nations Department of Economic and Social Affairs. URL: http://www.un.org/development/
desa/ (date of access: 30.01.2022). 12. United Nations Department of Economic and Social Affairs. URL: http://www.un.org/development/
desa/ (date of access: 30.01.2022). )
ономическое развитие Евразийского экономического союза и государств-членов в 2020 го
ународные рейтинги. URL: http://eec.eaeunion.org/upload/medialibrary/82a/Ekonomicheskoe 13. Экономическое развитие Евразийского экономического союза и государств-членов в 2020 го-
ду: международные рейтинги. URL: http://eec.eaeunion.org/upload/medialibrary/82a/Ekonomicheskoe-
razvitie-EAES-i-gosudarstv chlenov-v-2020-g.-Mezhdunarodnye-reytingi.pdf (дата обращения: 30.01.2022). 13. Экономическое развитие Евразийского экономического союза и государств-членов в 2020 го-
ду: международные рейтинги. URL: http://eec.eaeunion.org/upload/medialibrary/82a/Ekonomicheskoe-
razvitie-EAES-i-gosudarstv_chlenov-v-2020-g.-Mezhdunarodnye-reytingi.pdf (дата обращения: 30.01.2022). 14 UN E-Government Survey 2020 URL: http://publicadministration un org/egovkb/en-us/Reports/ у
ду
р д
р
p
g p
y
azvitie-EAES-i-gosudarstv_chlenov-v-2020-g.-Mezhdunarodnye-reytingi.pdf (дата обращения: 30. razvitie-EAES-i-gosudarstv_chlenov-v-2020-g.-Mezhdunarodnye-reytingi.pdf (дата обращения: 30.01.2022). 14. UN E-Government Survey 2020. URL: http://publicadministration.un.org/egovkb/en-us/Reports/
UN-E-Government-Survey-2020 (date of access: 30.01.2022). g
_
g
y
y
g p
(
р
)
14. UN E-Government Survey 2020. URL: http://publicadministration.un.org/egovkb/en-us/Report
UN-E-Government-Survey-2020 (date of access: 30.01.2022). 15. Данные, цифровизация и государственное управление. URL: http://www.vsemirnyjbank.org/
ru/region/eca/publication/europe-and-central-asia-economic-update (дата обращения: 30.01.2022). 15. Данные, цифровизация и государственное управление. URL: http://www.vsemirnyjbank.org/
ru/region/eca/publication/europe-and-central-asia-economic-update (дата обращения: 30.01.2022). g
p
p
p
(
р
16. Global Cybersecurity Index. URL: http://www.itu.int/en/ITU-D/Cybersecurity/Pag
ybersecurity-index.aspx (date of access: 30.01.2022). y
y
p (
)
17. Глобальный индекс кибербезопасности и профили по киберблагополучию. URL:
http://www.itu.int/dms_pub/itu-d/opb/str/D-STR-SECU-2015-PDF-R.pdf (дата обращения: 30.01.2022). 17. Глобальный индекс кибербезопасности и профили по киберблагополучию. URL:
http://www.itu.int/dms_pub/itu-d/opb/str/D-STR-SECU-2015-PDF-R.pdf (дата обращения: 30.01.2022). p
_p
p
p
(
р
)
18. Цифровой потенциал стран – участниц ЕАБР. URL: http://eabr.org/upload/iblock/551/
EABR_Digital_Potential_06_2019.pdf (дата обращения: 30.01.2022). 18. Цифровой потенциал стран – участниц ЕАБР. URL: http://eabr.org/upload/iblock/551/
EABR_Digital_Potential_06_2019.pdf (дата обращения: 30.01.2022). References Rating of the countries of the world according to the level of development of information and
communication technologies. Available at: http://gtmarket.ru/ratings/ict-development-index (accessed
30.01.2022) (In Russian). ) (
)
rnational Telecommunication Union (ITU). Available at: http://www.itu.int/ (accessed
2). 6. International Telecommunication Union (ITU). Available at: http://www.itu.int/ (accessed
30.01.2022). 6. International Telecommunication Union (ITU). Available at: http://www.itu.int/ (accessed
30.01.2022). 7 Rating of countries in the world according to the index of network readiness
Available at: )
Rating of countries in the world according to the index of network readiness. Available
tmarket.ru/ratings/networked-readiness-index (accessed 30.01.2022) (In Russian). )
7. Rating of countries in the world according to the index of network readiness. Av
ttp://gtmarket.ru/ratings/networked-readiness-index (accessed 30.01.2022) (In Russian). p
g
g
(
) (
)
8. Network Readiness Index. Available at: URL: http://networkreadinessindex.org/ (accessed 30.01.2022). 8. Network Readiness Index. Available at: URL: http://networkreadinessindex.org/ (accessed 30.01.2022). 9. Digital Adoption Index. Available at: http://www.worldbank.org/en/publication/wdr2016/Digital-
Ad
i
d
(
d 30 01 2022) 8. Network Readiness Index. Available at: URL: http://networkreadinessindex.org/ (accessed 30.01.2022). 9. Digital Adoption Index. Available at: http://www.worldbank.org/en/publication/wdr2016/Digital-
Adoption-Index (accessed 30.01.2022). p
g (
)
9. Digital Adoption Index. Available at: http://www.worldbank.org/en/publication/wdr2016/Digital-
Adoption-Index (accessed 30.01.2022). p
(
)
10. Digital Adoption Index (DAI): Measuring the Global Spread of Digital Technologies. Available at:
http://thedocs.worldbank.org/en/doc/587221475074960682-0050022016/original/WDR16BPDAImethodology.pdf
(accessed 30.01.2022). 10. Digital Adoption Index (DAI): Measuring the Global Spread of Digital Technologies. Available at:
http://thedocs.worldbank.org/en/doc/587221475074960682-0050022016/original/WDR16BPDAImethodology.pdf
(accessed 30.01.2022). (
)
11. Rating of the countries of the world according to the E-government development index. Available
at: http://gtmarket.ru/ratings/e-government-development-index (accessed 30.01.2022) (In Russian). (
)
11. Rating of the countries of the world according to the E-government development index. Available
at: http://gtmarket.ru/ratings/e-government-development-index (accessed 30.01.2022) (In Russian). 12. United Nations Department of Economic and Social Affairs. Available at: http://www.un.org/
development/desa/ (accessed 30.01.2022). 12. United Nations Department of Economic and Social Affairs. Available at: http://www.un.org/
development/desa/ (accessed 30.01.2022). 13. Economic development of the Eurasian Economic Union and member states in 2020: international
ratings. Available at: http://eec.eaeunion.org/upload/medialibrary/82a/Ekonomicheskoe-razvitie-EAES-i-
gosudarstv_chlenov-v-2020-g.-Mezhdunarodnye-reytingi.pdf (accessed 30.01.2022) (In Russian). 13. Economic development of the Eurasian Economic Union and member states in 2020: international
ratings. Available at: http://eec.eaeunion.org/upload/medialibrary/82a/Ekonomicheskoe-razvitie-EAES-i-
gosudarstv chlenov-v-2020-g.-Mezhdunarodnye-reytingi.pdf (accessed 30.01.2022) (In Russian). g
_
g
y
y
g p
(
) (
)
14. UN E-Government Survey 2020. Available at: http://publicadministration.un.org/egovkb/en-
us/Reports/UN-E-Government-Survey-2020 (accessed 30.01.2022). 14. UN E-Government Survey 2020. Available at: http://publicadministration.un.org/egovkb/en-
us/Reports/UN-E-Government-Survey-2020 (accessed 30.01.2022). 15. Data, digitalization and government. Available at: http://www.vsemirnyjbank.org/ru/region/
eca/publication/europe-and-central-asia-economic-update (accessed 30.01.2022) (In Russian). 15. References Data, digitalization and government. Available at: http://www.vsemirnyjbank.org/ru/region/
eca/publication/europe-and-central-asia-economic-update (accessed 30.01.2022) (In Russian). p
p
p
(
) (
)
16. Global Cybersecurity Index. Available at: http://www.itu.int/en/ITU-D/Cybersecurity/Pages/global-
cybersecurity-index.aspx (accessed 30.01.2022). 16. Global Cybersecurity Index. Available at: http://www.itu.int/en/ITU-D/Cybersecurity/Pages/global-
cybersecurity-index.aspx (accessed 30.01.2022). Òðóäû ÁÃÒÓ Ñåðèÿ 5 № 1 2022 12 Îïðåäåëåíèå ñòðàíîâûõ îñîáåííîñòåé öèôðîâèçàöèè â ãîñóäàðñòâàõ ÅÀÝÑ 17. Global Cyber Security Index and Cyber Wellbeing Profiles. Available at: http://www.itu.int/
dms_pub/itu-d/opb/str/D-STR-SECU-2015-PDF-R.pdf (accessed 30.01.2022) (In Russian). _p
p
p
(
) (
)
18. Digital potential of EDB member countries. Available at: http://eabr.org/upload/i
EABR_Digital_Potential_06_2019.pdf (accessed 30.01.2022) (In Russian). _p
p
p
(
) (
)
18. Digital potential of EDB member countries. Available at: http://eabr.org/upload/iblock/551/
EABR_Digital_Potential_06_2019.pdf (accessed 30.01.2022) (In Russian). Информация об авторах Новикова Ирина Васильевна – доктор экономических наук, профессор, заведующий кафедрой
менеджмента, технологий бизнеса и устойчивого развития. Белорусский государственный техноло-
гический университет (220006, г. Минск, ул. Свердлова, 13а, Республика Беларусь). E-mail:
xenia2012@belstu.by Новикова Ирина Васильевна – доктор экономических наук, профессор, заведующий кафедрой
менеджмента, технологий бизнеса и устойчивого развития. Белорусский государственный техноло-
гический университет (220006, г. Минск, ул. Свердлова, 13а, Республика Беларусь). E-mail:
xenia2012@belstu.by Равино Алла Васильевна – кандидат экономических наук, доцент, доцент кафедры менедж-
мента, технологий бизнеса и устойчивого развития. Белорусский государственный технологический
университет (220006, г. Минск, ул. Свердлова, 13а, Республика Беларусь). E-mail: ravino@belstu.by Равино Алла Васильевна – кандидат экономических наук, доцент, доцент кафедры менедж-
мента, технологий бизнеса и устойчивого развития. Белорусский государственный технологический
университет (220006, г. Минск, ул. Свердлова, 13а, Республика Беларусь). E-mail: ravino@belstu.by Information about the authors Novikova Irina Vasil’yevna – DSc (Economics), Professor, Head of the Department of Management,
Business Technology and Sustainable Development. Belarusian State Technological University (13a, Sverd-
lova str., 220006, Minsk, Republic of Belarus). E-mail: xenia2012@belstu.by Novikova Irina Vasil’yevna – DSc (Economics), Professor, Head of the Department of Management,
Business Technology and Sustainable Development. Belarusian State Technological University (13a, Sverd-
lova str., 220006, Minsk, Republic of Belarus). E-mail: xenia2012@belstu.by @
y
Ravino Alla Vasil’yevna – PhD (Economics), Associate Professor, Assistant Professor, the Department
of Management, Business Technology and Sustainable Development. Belarusian State Technological Uni-
versity (13a, Sverdlova str., 220006, Minsk, Republic of Belarus). E-mail: ravino@belstu.by
Поступила 14.02.2022
|
https://openalex.org/W2030998028
|
https://riunet.upv.es/bitstream/10251/140981/1/Cabrera%3bGomez%3bCabrera%20-%20Energy%20Assessment%20of%20Pressurized%20Water%20Systems.pdf
|
English
| null |
Energy Assessment of Pressurized Water Systems
|
Journal of water resources planning and management
| 2,015
|
cc-by
| 10,087
|
Introduction A growing population needs more water, more food, and therefore
more irrigation. Streamlining these processes is thus essential and
especially important in those agricultural countries that are shifting
from traditional to pressurized irrigation to save water. However,
this transformation process entails a heavy cost: namely, the energy
that pressurized water transport systems (PWTS) consume. In
California, such consumption accounts for up to 6% of the total
energy demand [Water in the West (WW) 2013]. In Europe, energy
consumption related to these uses is 109 TWh [Official Journal of
the European Union (OJEU) 2012]. Reducing or controlling this
consumption is crucial. More specifically, three new indicators are presented to achieve
this holistic efficiency assessment. The first two indicators are
based on an energy balance of the system, using the values of
the energy required by users, the minimum amount of energy to
be supplied to the system (because of an ideal behavior), and
the actual consumed energy. As a result, a real efficiency and an
ideal (and unachievable) efficiency are calculated. The third indi-
cator provides an achievable goal linked to a target level of energy
loss to account for inefficiencies in the system (e.g., pumping sta-
tions, leakage, friction loss). This goal is ambitious but achievable. The real efficiency indicator provides information on the actual
performance of the system, and when used in conjunction with the
ideal efficiency indicator it shows the maximum performance gap
that could be closed if efficiency was ideally improved. The ideal
level of efficiency is limited by the topographic energy (elevation
potential energy), and this depends on the topography of the
system. However, little attention has been given to a preliminary assess-
ment of the overall energy efficiency of PWTS. Improving pump
performance has always been important because of the steady rise
in energy costs (Perez-Urrestarazu and Burt 2012; OJEU 2012;
Papa et al. 2013). The efficient operation of pressurized systems
(Lingireddy and Wood 1998; Ulanicki et al. 2007; Giustolisi et al. 2013; Carriço et al. 2013) is also attracting considerable attention. Calculations of the energy embedded in water leaks (Colombo and
Karney 2002; Cabrera et al. 2010) and energy performance indica-
tors are being used to assess aspects of these systems (Pelli and Hitz
2000; Duarte et al. 2009). Energy Assessment of Pressurized Water Systems
E. Cabrera, M.ASCE1; E. G´omez2; E. Cabrera Jr.3; J. Soriano4; and V. Espert5 Energy Assessment of Pressurized Water Systems
E. Cabrera, M.ASCE1; E. G´omez2; E. Cabrera Jr.3; J. Soriano4; and V. Espert5 Abstract: This paper presents three new indicators for assessing the energy efficiency of a pressurized water system and the potential energy
savings relative to the available technology and economic framework. The first two indicators are the ideal and real efficiencies of the system
and reflect the values of the minimum energy required by users—the minimum amount of energy to be supplied to the system (because of its
ideal behavior) and the actual energy consumed. The third indicator is the energy performance target, and it is estimated by setting an
ambitious but achievable level of energy loss attributable to inefficiencies in the system (e.g., pumping stations, leakage, friction loss). The information provided by these three key performance indicators can make a significant contribution towards increasing system efficiency. The real efficiency indicator shows the actual performance of the system; the energy performance target provides a realistic goal on how the
system should be performing; and finally, the ideal efficiency provides the maximum and unachievable level of efficiency (limited by the
topographic energy linked to the network topography). The applicability and usefulness of these metrics will be demonstrated with an ap-
plication in a real case study. DOI: 10.1061/(ASCE)WR.1943-5452.0000494. This work is made available under the terms of the Creative
Commons Attribution 4.0 International license, http://creativecommons.org/licenses/by/4.0/. improvement margins, which is precisely the main contribution
of this paper. 1Professor, ITA, Universitat Politècnica de València, Apdo, 22012
Valencia, Spain.
2Ph.D. Student, ITA, Universitat Politècnica de València, Apdo, 22012
Valencia, Spain.
3Associate Professor, ITA, Universitat Politècnica de València, Apdo,
22012 Valencia, Spain.
4Assistant Professor, ITA, Universitat Politècnica de València, Apdo,
22012 Valencia, Spain (corresponding author). E-mail: jasool@ita.upv.es
5Professor, ITA, Universitat Politècnica de València, Apdo, 22012
Valencia, Spain.
Note. This manuscript was submitted on January 10, 2014; approved on
October 3, 2014; published online on November 6, 2014. Discussion period
open until April 6, 2015; separate discussions must be submitted for indi-
vidual papers. This paper is part of the Journal of Water Resources Plan-
ning and Management, © ASCE, ISSN 0733-9496/04014095(12)/$25.00. J. Water Resour. Plann. Manage. Note. This manuscript was submitted on January 10, 2014; approved on
October 3, 2014; published online on November 6, 2014. Discussion period
open until April 6, 2015; separate discussions must be submitted for indi-
vidual papers. This paper is part of the Journal of Water Resources Plan-
ning and Management, © ASCE, ISSN 0733-9496/04014095(12)/$25.00. 1Professor, ITA, Universitat Politècnica de València, Apdo, 22012
Valencia, Spain.
2Ph.D. Student, ITA, Universitat Politècnica de València, Apdo, 22012
Valencia, Spain.
3Associate Professor, ITA, Universitat Politècnica de València, Apdo,
22012 Valencia, Spain.
4Assistant Professor, ITA, Universitat Politècnica de València, Apdo,
22012 Valencia, Spain (corresponding author). E-mail: jasool@ita.upv.es
5Professor, ITA, Universitat Politècnica de València, Apdo, 22012
Valencia, Spain. Introduction However, no one has yet proposed met-
rics that provide a global view of system efficiency and existing Once a diagnostic is known and providing improvement
margins are significant—then a second stage can be considered. This second phase consists in analyzing where energy is lost by
means of a system audit (Cabrera et al. 2010). This audit enables
determining final uses with enough precision to identify the largest
inefficiencies (e.g., leakage, friction, pumps) and the largest poten-
tial savings. With the global analysis completed, the focus shifts to
those aspects or solutions with the greatest cost-benefit relation
(think globally, act locally). The second stage is unnecessary if there
is little difference between the efficiency target and the diagnostic. Downloaded from ascelibrary.org Last, the possibility of recovering part of the topographic en-
ergy of the system (to be defined later) should be explored. This
may be achieved by installing pumps to work as turbines (PATs)
(Carravetta et al. 2012), or by dissipating energy with pressure reduc-
ing valves (PRVs). The latter practice is more widespread—but
both approaches work by reducing the excess pressure in the fluid’s
energy to decrease water leaks and pipe stress. Note. This manuscript was submitted on January 10, 2014; approved on
October 3, 2014; published online on November 6, 2014. Discussion period
open until April 6, 2015; separate discussions must be submitted for indi-
vidual papers. This paper is part of the Journal of Water Resources Plan-
ning and Management, © ASCE, ISSN 0733-9496/04014095(12)/$25.00. This paper presents new assessment concepts with the corre-
sponding metrics. The presented improvement process takes the
status quo diagnostic (real efficiency of the present state) as a tool
to target specific efficiency gains with current technologies while 04014095-1 © ASCE J. Water Resour. Plann. Manage. Ideal System without Excess Pressure In an ideal frictionless system and for any period of time, the
hydraulic grade line (HGL), sum of the natural (or gravitational)
head, hni, and the pump head, hpi, (or total dynamic head,
TDH) is constant and equal to Hhi ¼ Hji ¼ Hli. However, the total
supplied energy is proportional to the corresponding demand of
each period. An ideal system is one where there are no friction head losses or
leaks (while the kinetic energy is disregarded, a common practice in
network analysis). There is no excess pressure because at the criti-
cal point, pci, is equal to the required service pressure, p0. In this
case, the critical point pressure is also the required pressure (Fig. 1),
and then pci ¼ phi ¼ p0. But it must be underlined that in real
cases this may not be the case. In a real system, friction losses depend on flow and leakage
rates. As a result, the supplied head (usually the pump head) must
be adjusted to meet the steady demand of each period. This implies
that a control system [such as a variable frequency drive (VFD)]
must be present at the pumping station. Energy balance calculations
are mathematically static and then only valid during brief periods of
time (Δt). Therefore, for longer time periods (h, day, month or
year), the total supplied energy must be calculated by integration
(Cabrera et al. 2010)—meaning an extended period simulation
(Rossman 2000). Alternatively, this energy can be assessed by
adding the wire energy (electricity bill) and gravitational energy,
the latter being γhniðV þ ΔVÞ, assuming a constant suction water
level (zn). The application of the continuity equation to the system shown
in Fig. 1 (and for a given steady flow time period) is straight-
forward. The total injected volume at the pumping station (V) is
equal (in the absence of water losses) to the sum of the demands
of all the consumption nodes (V ¼ P vj). The supplied energy in
that period is Esi ¼ γVHhi ¼ γ
X
vj
ðzj −zlÞ þ pji
γ
¼ γ
X
vj
ðzj −zlÞ þ p0
γ þ ðzh −zjÞ
¼ Euo þ Eti
ð1Þ Finally, it is emphasized that, in this ideal case, the type of
supplied energy (hpi or hni) is irrelevant for assessing an energy
balance. J. Water Resour. Plann. Manage. Fig. 1. Ideal pressurized water system (partial profile without excess energy) nica De Valencia on 11/07/14. Copyright ASCE. For personal use only; all rights reserved. consi
Cabr
Bas
Fig. most
the h
and t
zh ¼
form
node
for g
Idea
An i
leaks
netw
cal p
case,
and
cases
T
in Fi
forw
equa
of al
that p
wher
to al
Downloaded from ascelibrary.org by Universidad Politecnica De Valencia on 11/07/14. Copyright ASCE. For personal use only; all rights reserved. Fig. 1. Ideal pressurized water system (partial profile without excess energy) considering both water and energy costs. This paper simplifies
Cabrera’s six-step process to three steps (Cabrera et al. 2014). horizontal line and the dashed line). In a flat network, Eti would
be zero. This energy also represents the amount of energy that, in
theory, could be recovered by installing PATs at every consumption
node. The energy to be recovered in Δt at any demand node by the
corresponding PAT is vj (the volume consumed at node j in the
considered time period) times the available topographic pressure,
pjt;i. In reality, recovering all this energy is impossible. In many
systems, this excess (when considered from the user’s perspective)
energy is dissipated with PRVs (see “Dissipation and Recovery of
Energy (with PRV or PATs) in a PWTS”). Ideal System without Excess Pressure After all, every single kWh is the same from a physical
perspective, regardless of its origin. where Hhi is the piezometric head at the highest node (ideally equal
to all nodes). Additionally Euo ¼ γ
X
vj
ðzj −zlÞ þ p0
γ
Eti ¼ γ
X
vjðzh −zjÞ ¼ γ
X
vj
pjt;i
γ
ð2Þ Basic Concepts Fig. 1 represents a partial profile view of a PWTS that shows the
most characteristic points of the system: the pumping station;
the highest and lowest nodes (zh and zl); a generic node (zj);
and the lowest pressure node, called critical (zc). In an ideal system,
zh ¼ zc. To establish the energy balance that enables defining per-
formances and assessing the overall system energy efficiency, the
node with the lowest elevation of the system is chosen as the origin
for gravitational energies (Cabrera et al. 2010) and then zl ¼ 0. When the pressure at the critical point is as required (as detailed
in Fig. 1), the supplied energy is the same as the minimum required
energy or base energy Ebi. Therefore Esi ¼ Euo þ Eti ¼ Ebi
ð3Þ ð3Þ J. Water Resour. Plann. Manage. 04014095-2 J. Water Resour. Plann. Manage. J. Water Resour. Plann. Manage. J. Water Resour. Plann. Manage. Ideal System with Excess Pressure ð2Þ The only difference between the systems in Figs. 1 and 2 is that in
the second case the supplied energy (pressure) is above the mini-
mum necessary value. Fig. 2 shows a pressure value at the critical
point higher than the required, po (pei represents excess pressure). To distinguish this case from the previous case, the pressure and
head terms are represented with an asterisk. being γ the specific weight of water, Euo the minimum energy
required by the users, and Eti the topographic energy (elevation
potential energy) of the system. This latter term depends on the
terrain’s irregularities, thus its suggested name. In Fig. 1, the
shaded area is proportional to Eti (the area between the top © ASCE
04014095-2 © ASCE
04014095-2 © ASCE J. Water Resour. Plann. Manage. 04014095-2 Real System The energy to be supplied, Euo, remains constant (Fig. 3). However,
the topographic energy of the system, Etr, (shaded area) changes
because it is linked to the hydraulic head line. Energy losses, which
are referred to as global reducible energy, Erg, are now different
from zero, and include: Esr ¼ Euo þ Etr þ Erg þ Eer
¼ Euo þ Etr þ Erg þ γðV þ ΔVÞ
pcr −p0
γ
ð7Þ Esr ¼ Euo þ Etr þ Erg þ Eer ¼ Euo þ Etr þ Erg þ γðV þ ΔVÞ
pcr −p0
γ
ð7Þ ð7Þ •
Energized water in leaks (Erl). Apparent losses, such as
metering inaccuracies, must be included, giving rise to apparent
system energy inefficiency; •
Energized water in leaks (Erl). Apparent losses, such as
metering inaccuracies, must be included, giving rise to apparent
system energy inefficiency; with ΔV being the total water losses in the system, and Eer the
surplus energy (which can be significant because the whole input
volume to the system is subject to this excess pressure). •
Energy dissipated by friction (Erf) in pipes, valves, and other
elements; J. Water Resour. Plann. Manage. Fig. 2. Ideal pressurized water system (partial profile with excess energy) Fig. 2. Ideal pressurized water system (partial profile with excess energy) Fig. 3. Real pressurized water system (partial profile without excess energy) Fig. 3. Real pressurized water system (partial profile without excess energy) The higher pressure (p
hi > po) is attributable to excess energy
entering the system In summary, the real energy supplied to the system is In summary, the real energy supplied to the system is Esr ¼ Euo þ Etr þ Erg ¼ ðEuo þ EtrÞ þ ðErl þ Erf þ Erp þ EroÞ
ð6Þ pei ¼ p
hi −po;
Eei ¼ γV pei
γ
ð4Þ ð4Þ ð6Þ In a real system, however, the minimum required energy or base
energy depends on its hydraulic operation and, therefore, defining a
term equivalent to Ebi makes no sense. However, Ebi will still re-
present the lower limit of this base energy, which corresponds to the
case of a real system in which losses tend to zero. It is also noted
that the hydraulic head is no longer a straight line because head
losses are not uniform. In this case, the system’s input energy is Esi ¼ Euo þ Eti þ Eei
with Esi > Ebi
ð5Þ ð5Þ Eti is obviously the same as in the previous case (and propor-
tional to the shaded area), whereas the excess energy, Eei, (avoid-
able most of the time) must be corrected with operational measures. Eq. (6) should include the surplus energy if the pressure at the
critical point (the point with the lowest pressure in the network)
exceeds the required pressure. Then 04014095-3 J. Water Resour. Plann. Manage. J. Water Resour. Plann. Manage. Energy Efficiency of a Pressurized Water System •
Energy loss in pumping stations (Erp) attributable to several
inefficiencies (electrical, friction, pump, and operational losses);
and From the concepts and the energy balance presented in the previous
section, the energy efficiency of PWTS can be stated as the relation •
Other losses (Ero) such as break pressure tanks. 14095-3
J. Water Resour. Plann. Manage. 04014095-3 Energy Efficiency Target In an ideal system, all the topographic energy is recovered, and
the total useful energy will be the sum of Euo and Eti, whereas an
ideal performance will be equal to By comparing the ideal performance (ηai) with the real perfor-
mance (ηar) [Eqs. (9) and (10)], the difference ηai −ηar provides
an initial estimation of the system’s improvement gap. Because this
diagnostic has to be referred to the optimum value, ηai, this value
must be calculated by making Eei ¼ 0 (an unnecessary energy that
should always be zero). ηar has already been calculated, and ηai can
be easily determined from Eqs. (1), (2), and (9). ηwi ¼ Euo þ Eti
Esi
¼ 1 −Eei
Esi
ð8Þ ð8Þ ð8Þ In this case, the only possible inefficiency can be the result of an
excessive energy supply [Eei, Eq. (4)]. If Eei ¼ 0, performance
would be one (ηwi ¼ 1), a utopian value obtained from the ideal
assumptions made. Once the (ηai −ηar) difference is known, the next step is deter-
mining the potential improvement gap. Or in other words, how
close the second term can get to the first term through improvement
measures while maintaining an acceptable cost-benefit ratio. A new
reference value (ηar;o) represents the achievable system efficiency,
or target efficiency, that verifies ηai > ηar;o > ηar. This value can
be estimated from Eq. (10) assuming that Eer ¼ 0. The numerator
is a system invariant, whereas the denominator is given by Eq. (6)
(with the recovered energy being zero). Minimizing the energy
losses implies reducing as much as possible the four reducible
terms (Erl, Erf, Erp and Ero) and understanding which fraction
of the topographic energy (Etr) can be reduced. Therefore, an ef-
ficiency target can be identified for all five terms as Etr;o, Erl;o,
Erf;o, Erp;o, and Ero;o. From all these estimated values, and taking
into account Eq. (7), Esr;o will result In reality, Eti is (partially or totally) lost because energy recov-
ery only makes economic sense in a few occasions. The most
common scenario is one where no recovery exists. In this case,
the system efficiency, ηai, is equal to ηai ¼ Euo
Esi
¼ 1 −Eti
Esi
−Eei
Esi
¼ 1 −θti −Eei
Esi
ð9Þ ð9Þ If no excess energy is supplied, then Eei ¼ 0, ηai and θti are
complementary with a sum equal to 1. J. Water Resour. Plann. Manage. ð11Þ ηar ¼ 1 −ðθtr þ θer þ λrl þ λrf þ λrp þ λroÞ between the minimum energy required by users and the actual sup-
plied energy. To introduce these concepts progressively, the ideal
case is analyzed first. Analyzing the efficiency of a system should start by calculating
ηar. The numerator (Euo) is known, and the denominator (Esr) is
calculated, as said before, from the energy consumed by the pump
(e.g., value from the electricity bill) plus the gravity (natural) sup-
plied energy that is dependent on the characteristics of the system. The global value (Esr) is then known, but not its components [sec-
ond part of Eq. (11)]. Determining their values requires a system
audit. The best strategies to improve performance can be selected
once the contribution of each term is known. Energy Efficiency of an Ideal System (with and without
Energy Recovery) The minimum required energy (both in real and ideal systems) is
Euo, whereas the supplied energy in an ideal system is Esi. If a part
of the topographic energy included in Esi is recovered, the energy
efficiency of the PWTS will improve. Therefore, it seems reason-
able to define two different ratios depending on this fact, ηwi (with)
and ηai (without recovery). ica De Valencia on 11/07/14. Copyright ASCE. For personal use only; all rights reserved. Energy Efficiency Target The topographic parameter
represents the fraction of the supplied energy that is lost (assuming
that no energy is recovered) attributable to the system excess pres-
sure (topographic pressure, pjt;i, Fig. 1). If θti approaches its upper
limit (a system with an irregular topography), then installing PRVs
to reduce pressure levels should be considered to minimize leaks
and reduce stress in the pipes. In such cases, however, the possibil-
ity of introducing physical changes in the system to reduce Eti
(dividing the system into different pressure areas) should have been
considered previously. In ideal flat networks, θti is zero, and then
ηwi is irrelevant. Esr;o ¼ ðEuo þ Etr;oÞ þ Erg;o
¼ ðEuo þ Etr;oÞ þ ðErl;o þ Erf;o þ Erp;o þ Ero;oÞ
ð12Þ ð12Þ As more system inefficiencies are removed, Esr;o will approach
Esi [Eq. (1)]. In Eq. (12), the first term Euo is a system invariant (see
“Synthesis of the Assessment”), whereas the real topographic
energy (Etr;o) depends on the new level of losses and, there-
fore, on the new pressures calculated for the more efficient
scenario Overall, these performances confirm an obvious truth: when en-
ergy losses are zero, supplying the minimum energy (Ebi) and
recovering all the topographic energy (Eti) results in a performace
value of 1 [Eq. (8)]. Without topographic energy recovery and with
Eei ¼ 0, the maximum performace is one minus the fraction of
topographic energy in the system [Eq. (9)]. Downloaded from ascelibrary.org b Etr;o ¼ γ
X
vj
pjt;r
γ
o
ð13Þ ð13Þ J. Water Resour. Plann. Manage. Energy Efficiency of a Real System The pressures values in the network can be determined with a
mathematical model adjusted for the new level of losses. This will
enable the complementary pressures ðpjt;rÞo to be obtained. Eti can
be directly calculated from Eq. (2). Both values are very similar,
and so it can be assumed that Analyzing ideal systems enables establishing maximum values for
system performances. Real systems share with ideal systems the
numerator of the energy efficiency (Euo), whereas the denominator
changes to include energy losses. Then Eq. (7) becomes Etr;o −Eti ¼ γ
X
vj
pjt;r
γ
o
−pji;r
γ
→0
ð14Þ ηar ¼ Euo
Esr
¼ 1 −Etr
Esr
−Eer
Esr
−Erg
Esr
¼ 1 −θtr −θer −λrg
ð10Þ ð14Þ ð10Þ where energies and reducible losses are expressed on a per unit
basis (θ for the energies and λ for reducible losses). When these
losses are broken into different terms, the contribution of each
to the system inefficiencies can be clearly seen with the expression This term will have a positive or negative sign depending on the
system topography, the position of the critical point, and the work-
ing conditions. In a flat network, the difference will always be pos-
itive. To provide service pressure to the furthest node (in this case, 014095-4
J. Water Resour. Plann. Manage. © ASCE 04014095-4 04014095-4 J. Water Resour. Plann. Manage. Reducible Energy Attributable to Friction Losses, Erf ;o ;
Taking all this into account Taking all this into account This is the most uncertain estimation, especially in large systems
with loops, redundancies, and parallel flows. It requires assigning
an average energy loss, related to an average path for the total mo-
bilized volume of water. This path, dependent on the demand and
leak distributions, is very variable. Consumptions which are very
close to supply points barely have losses. On the other hand, those
further away have significant losses. An average path Lpm must be
estimated between the pumping station and the consumption nodes,
and an average unit head loss (Jm) (m=km) for all pipes. Again, this
is not an easy estimation. Jm is a function of the square value of the
flow rate (variable in time with demand and leakage). The preced-
ing factors represent a significant risk of making a biased assess-
ment of this factor. Esr;o ¼ ðEuo þ Etr;oÞ þ Erg;o
≈ðEuo þ EtiÞ þ ðErl;o þ Erf;o þ Erp;o þ Ero;oÞ
ð15Þ ð15Þ And therefore ηar;o ¼ Euo
Esr;o
¼ Euo
Esi
Esi
Esr;o
¼ ηai
Esi
Esr;o
ð16Þ ð16Þ The target efficiency ηar;o, depends on the objective level of
energy losses. The criteria used to assess these acceptable levels
of energy losses follow below. Additionally, local head losses (Δprf;p) must be added owing to
the presence of filters, valves, and manifolds. With these assump-
tions, the following relationship results: Reducible Energy Embedded in Leaks, Erl;o This reduction requires recovering the total volume of reasonable
leaks (ΔVo), the sum of the losses (Δvjo) at each node for a given
time period. The energy loss embedded in leaks would then be Erf;o ¼ γðV þ ΔVoÞ
ðLpmÞJm þ Δprf;p
γ
ð18Þ ð18Þ Erl;o ¼ γ
X
Δvjo
pjr
γ ¼ γ
X
Δvjo
po
γ þ pjt;r
γ
≈γΔVo
po
γ þ
zh −zh þ zl
2
ð17Þ Friction head losses in PRVs will be considered later (see
following section). ð17Þ J. Water Resour. Plann. Manage. Fig. 4. Etr;o and Eti comparison profiles for: (a) system a; (b) system b nica De Valencia on 11/07/14. Copyright ASCE. For personal use only; all rights reserved. Downloaded from ascelibrary.org by Universidad Politecnica De Valencia on 11/07/14. Copyright ASCE. For personal use only; all rights reserved. Fig. 4. Etr;o and Eti comparison profiles for: (a) system a; (b) system b the critical node), the head pressure must be greater than the ideal
required pressure. In a system with irregular terrain, this value will
depend on the topography and the position of the critical point. In
Fig. 4, system (a) has a negative difference [Eq. (14)], whereas
system (b) is positive. However, these differences will always be
small. Therefore, it is reasonable to assume Etr;o ≈Eti. in pipes attributable to higher circulating flow rates as a result
of leakage. Eq. (17) is suitable for the estimation of Erl;o (to
calculate later ηar;o). J. Water Resour. Plann. Manage. 04014095-5 04014095-5 Reducible Energy in Pumping Stations, Erp;o This approximation implies concentrating all the leaks in a
median pressure node. Its value can be estimated assuming an ideal
behavior of the system (pjt;i=γ ¼ zh −zj, Fig. 1). This approxima-
tion is valid for losses of approximately 10–15%, but for higher
ratios it could introduce a bias in the calculations. For this paper’s
purposes, this is not a problem, and the level of losses must be low
to set a target. In any case, the exact calculation of the energy em-
bedded in the leaks requires an energy audit (Cabrera et al. 2010),
because Eq. (17) does not consider the additional friction losses Pumping stations are usually responsible for a high percentage
of energy losses. However, estimating a reasonable value is not
difficult (using the average combined efficiency (ηpo) of the
variable frequency drive/motor/pump and the corresponding
pump head). When the pump’s head-flow curve is available, these
losses are easy to calculate. Otherwise, they can be estimated
using Eq. (19), in which Lpc is the distance between the pumping
station and the critical point, whereas zn represents the pump
suction level © ASCE J. Water Resour. Plann. Manage. Synthesis of the Assessment Excess pressure in the network may produce a considerable
number of leaks (Giustolisi et al. 2008). To reduce these leaks,
PRVs are usually installed to limit pressure at consumption nodes
to a service value po. In this case, part of Etr is dissipated at the
valves. However, although energy is dissipated at the PRVs, the
total energy demanded by the system diminishes because the vol-
ume of leaks is smaller—reducing the consumed volume, the flow
rate, and the friction losses. In short, Esr decreases because Erl and
Erf decrease. Energy dissipated at the PRVs is at the expense of Etr
and which, as seen before in Eq. (14) and Fig. 4, can be assumed
constant provided the required pressure (po) is satisfied. For this
reason, as Fig. 5 shows, these losses in PRVs do not affect ηar;o
and only change ηwr. The evaluation of the system efficiency is based on three indicators:
ηai [Eq. (9)]; ηar [Eq. (10)]; and ηar;o [Eq. (16)]. These are defined
from four terms: Esi [Eq. (1)]; Euo [Eq. (2)]; Esr or sum of the wire
(electricity bill)—and gravitational energies supplied to the system
and Esr;o [Eq. (15)]. All refer to the control volume (CV) previously
selected and integrated over significant periods of time to facilitate
the Esr calculation (e.g., a month, quarter, or year). Any CV can be
considered as long as the mass and energy flows through the boun-
dary surface (control surface, CS) are known during the specified
period of time. This provides the global energy assessment for the
system in time and space. The weakest point of this analysis, the target energy estimation,
Esr;o (and therefore, ηar;o) has little relevance to the whole process
because the value (ηar;o −ηar) is only used to trigger the next step
(the analysis phase with the corresponding audits, Fig. 5). From an
engineering point of view, some subjective estimations and sim-
plifications required to determine Esr;o (see “Energy Efficiency
Target”) have little influence on the results of the analysis. The
key factor in the analysis is the order of magnitude of the term
(ηar;o −ηar), a value that, in absolute terms, is not especially sen-
sitive to the formulated hypotheses. In any case, these estimated
terms can be precisely determined from the energy audit (Cabrera
et al. 2010), and this value can therefore be determined during the
second phase. Other Reducible Energy Losses, Ero;o In one of the paths, the efficiency of the real system (ηar) is
estimated and compared with the target performance value,
ηar;o. At the same time, θti is calculated. If this value is relevant
(i.e., θti > 0.2), the possibility of reducing it through subdividing
the system should be explored. This strategy has been applied to the
case study which is a tree-like irrigation network. In fact, one of the
major improvements is subdividing the system into three areas with
a higher, medium, and lower elevation, which implies reducing the
global θti. Further losses can be found in PWTS. Break pressure tanks (cham-
bers to avoid depressions in high points are required in pipes with
an irregular profile) can be responsible for important energy losses
that should be avoided—similar to an excess of energy (Eer) in-
jected in the system. Break pressure tanks (e.g., domestic tanks)
are equally inconvenient; in such cases the value of the wasted en-
ergy (Ero) can be calculated by multiplying the depressurized vol-
ume times the network pressure at the delivery node. In the absence
of inefficiencies If the economic analysis does not enable reducing the system’s
θti (energy savings are insufficient to justify the required invest-
ment) and if its value is relevant enough (i.e., θti > 0.2) the use
of both PRVs and PATs in the system should be studied. A
cost-benefit analysis would then indicate which solution is better. If some turbines are finally installed, the system’s efficiency will be
assessed with ηwr (which takes into account the recovered energy). If the economic analysis does not enable reducing the system’s
θti (energy savings are insufficient to justify the required invest-
ment) and if its value is relevant enough (i.e., θti > 0.2) the use
of both PRVs and PATs in the system should be studied. A
cost-benefit analysis would then indicate which solution is better. If some turbines are finally installed, the system’s efficiency will be
assessed with ηwr (which takes into account the recovered energy). A satisfactory diagnostic requires no further action. If additional
efficiencies can be gained, the analysis of possible improvement
measures and their implementation is time consuming. In a con-
tinuous improvement process, an efficiency improvement cycle
may take several months; achieving significant efficiencies will
normally take at least one year. Ero;o ¼ 0
ð20Þ ð20Þ These estimations enable calculating the performance reference
value (ηar;o) [Eqs. Other Reducible Energy Losses, Ero;o (15) and (16)]. A satisfactory diagnostic requires no further action. If additional
efficiencies can be gained, the analysis of possible improvement
measures and their implementation is time consuming. In a con-
tinuous improvement process, an efficiency improvement cycle
may take several months; achieving significant efficiencies will
normally take at least one year. Dissipation and Recovery of Energy (with PRV or
PATs) in a PWTS The topographic pressure line (pjt;i and pjt;r) created by land
irregularities is an unavoidable factor (Figs. 1 and 3) that leads
to higher pressures in the system. However, other pressure sur-
pluses, such as pei and per, are avoidable in most cases and will
not be considered from this point onwards. Improving the Efficiency of a PWTS ð19Þ Fig. 5 synthesizes the energy efficiency improvement process in a
PWTS. This flow chart has two columns or paths that, to some ex-
tent, are simultaneously decoupled and complementary. J. Water Resour. Plann. Manage. Erp;o ¼ γðV þ ΔVoÞhpr
1
ηpo
−1
¼ γðV þ ΔVoÞ
LpcJm þ Δprf;p
γ
þ po
γ þ ðzc −znÞ
×
1
ηpo
−1
ð19Þ recovery stations improves the overall energy efficiency of the sys-
tem. In this case, ηwr should be used as a performance measure
instead of ηar. J. Water Resour. Plann. Manage. J. Water Resour. Plann. Manage. Synthesis of the Assessment However, from an energy point of view, it is much better to re-
cover energy installing turbines, or PATs than to dissipate energy
with PRVs. An initial approach to identify candidate pipes would
be to multiply the average flow at line k, qk by the average excess
pressure
at
the
end
node
j,
ðpjr −poÞ. Pipes
verifying
qk ðpjr −poÞ > Pmin would be selected as potential candidates
for a recovery station. The recovered energy (part of the initial
topographic energy) is termed Eyr (Fig. 5). y
The next stage consists in deciding the regulation system
(Carravetta et al. 2014) that enables the recovery of the maximum
amount of energy without compromising the service pressure (po)
at the consumption nodes downstream of the installation. Only the
recoverable amount of energy (Eyr) can be calculated. However, in
practice, just a small part of Etr is recoverable. This global assessment is based on average values and cannot
reflect any temporal evolution of the system losses nor the influ-
ence of their location. This must be taken into account, particularly
if during the considered period the pumps are working under
variable flow conditions. In such cases, maintaining high pump The use of turbines instead of PRVs has become more com-
mon in recent years (Fontana et al. 2012) and should become
an even more feasible option in the future because including energy 04014095-6 © ASCE J. Water Resour. Plann. Manage. J. Water Resour. Plann. Manage. Fig. 5. Flowchart to improve the efficiency in PWST Fig. 5. Flowchart to improve the efficiency in PWST •
Water and energy flows coming into the CV must be known. efficiencies at all times becomes a difficult task; an issue that, be-
cause of its relevance, has been on the research agenda for many
years (Ormsbee et al. 1989; Walski 1993; Ulanicki et al. 2007; Papa
et al. 2013; Kurek and Ostfeld 2014). •
Water delivered to users and leaving the CV must be known. •
The elevation of system nodes must be known and pressure (po)
defined. In practice this pressure standard should not only be
defined but also actually met. Downloaded from ascelibrary.org In the assessment hereby proposed, the target value for the re-
quired energy in pumping stations is estimated by setting an ambi-
tious although realistic average efficiency target. J. Water Resour. Plann. Manage. Synthesis of the Assessment The presented
case study corresponds to an on-farm irrigation schedule with a
constant pumping flow rate. •
If no minimum pressure standard is defined, po should be the
minimum pressure at the critical node. •
The zero value for node elevation will be assigned to the lowest
node (zl ¼ 0). Potential energy terms will be referred to this
lowest node. In summary, the proposed assessment provides a global indica-
tion of the system efficiency, but with no information on where and
how energy is lost. Structural inefficiencies (owing to inappropriate
designs), and those resulting from operational decisions are all in-
cluded in ηar. •
Wire energy consumption for the system must be known. gy
p
y
From this data, ηai and ηar can be calculated, whereas ηar;o, will
be set as the target reference. Using these hypotheses to set this
target comes at the cost of some uncertainty. However, although
this target may not be considered a fixed reference, it provides a
goal that is close enough to the real goal and helps generate change
and efficiency. A similar example can be found in water loss, where
the infrastructure leakage index (ILI) (Lambert et al. 1999) provides
a far from accurate target, and yet it has changed the international
scene by helping to promote the reduction of leakage around the
world. In this particular case, the target is the result of a conscious
decision at the desired level of service—promoting a more active This simple assessment (which can be carried out with a simple
spreadsheet) will identify the presence (or absence) of an energy
problem in a system. To make the assessment: •
A system-wide CV must be defined. The same approach could
be used to analyze subsystems (through alternative CVs). In
such cases, these subsystems or district energy areas (DEAs)
can play (in energy terms) a similar role to DMA (district me-
tering areas). •
A system-wide CV must be defined. The same approach could
be used to analyze subsystems (through alternative CVs). In
such cases, these subsystems or district energy areas (DEAs)
can play (in energy terms) a similar role to DMA (district me-
tering areas). © ASCE 04014095-7 J. Water Resour. Plann. Manage. J. Water Resour. Plann. Manage. J. Water Resour. Plann. Manage. Fig. 6. Network layouts with three energy sources: (a) case a—natural and wire energy are supplied to the system; (b) case b—only wire energy is
supplied to the system; (c) case c—only natural energy is supplied to the system Fig. 6. Network layouts with three energy sources: (a) case a—natural and wire energy are supplied to the system; (b) case b—only wire energy is
supplied to the system; (c) case c—only natural energy is supplied to the system is the subject of this paper. The results that follow have been
confirmed in practice. participation in target setting—whereas ILI coefficients have a set
value that cannot be changed. p
Fig. 7 depicts a real system with more than 400 consumption
nodes and 55 km of pipes. The diversity of situations to be found in practice is enormous. Fig. 6 depicts three typical network layouts with different energy
inputs. In case (a), similar to the example that follows, wire and
natural energy are supplied to the system. The compensation tank
acts as a water and energy flywheel (Cabrera et al. 2010). In case
(b), only wire energy is supplied to the system because the lowest
node is situated at the water table level (at the borehole). In case (c),
the potential is the only source of energy. In this latter system, the
installation of a PRV will introduce friction losses in the system, but
has no influence on the efficiency of the system because the losses
generated by the PRV are at the expense of topographic energy
(Fig. 5, right side). The assessment presented here was prepared with data
from 2011: •
Irrigation ≡125.51 equivalent days per year, a value that
depends on yearly rainfall. It works in the same way as an inter-
mittent urban supply. Fig. 7. Cap de Terme irrigation network Case Study This case study corresponds to a real pressurized irrigation network
(Cap de Terme) that has been operational since 2006. The need for
the study arose in 2008 when energy for agricultural use lost all
subsidies, and the electricity bills paid by farmers significantly in-
creased in just two years. Most of the ideas in this paper were in-
spired from this study in a bottom up process. Although the actual
study has already been concluded (see the different stages in Fig. 5),
the analysis that follows only contains the diagnostic stage, which Fig. 7. Cap de Terme irrigation network © ASCE © ASCE 04014095-8 J. Water Resour. Plann. Manage. J. Water Resour. Plann. Manage. Esi ¼ γ
X
vj
ðzh −zlÞ þ p0
γ
¼ γV
po
γ þ ðzh −zlÞ
Esi ¼ γ
X
vj
ðzh −zlÞ þ p0
γ
¼ γV
po
γ þ ðzh −zlÞ
¼ 2,082.94 kWh=day
Euo ¼ γ po
γ V þ γ
X
vjðzj −zlÞ ¼ 1,494.23 kWh=day
Eti ¼ γ
X
vj
pjt;i
γ
¼ γ
X
vjðzh −zjÞ ¼ 588.71 kWh=day Downloaded from ascelibrary.org by Universidad Politecnica De Valencia on 11/07/14. Copyright ASCE. For personal use only; all rights reserved. For ideal and real energy intensities (Iei and Ier, respectively),
the relation remains the same Eti ¼ γ
X
vj
pjt;i
γ
¼ γ
X
vjðzh −zjÞ ¼ 588.71 kWh=day Iei ¼ Esi
V ¼ 2,082.94
18,580 ¼ 0.11 kWh=m3
Ier ¼ Esr
V ¼ 6,497
18,580 ¼ 0.35 kWh=m3 The theoretical value of the achievable performance is ηai ¼ Euo
Esi
¼ 1,494.23
2,082.94 ¼ 0.72 Ier ¼ Esr
V ¼ 6,497
18,580 ¼ 0.35 kWh=m3 The percentage of the topographic energy is, logically, the com-
plementary value If, as usual, energy intensities are only referred to wire energy
(ignoring the supplied natural energy) the performance indicator,
logically, improves. In this case θti ¼ Eti
Esi
¼ 588.71
2,082.94 ¼ 0.28 I 0er ¼ Esr;p
V
¼ 5,833
18,580 ¼ 0.31 kWh=m3 The real value of ηar is determined from registered energy con-
sumptions. Therefore ηar ¼ Euo
Esr
¼ 1,494.23
6,497
¼ 0.23 However this analysis fails to accurately reproduce reality. If all
the energy was natural, this value would be zero, an unacceptable
result from a physical point of view, although system efficiencies
are less relevant in practice. In such a case, it would only make
sense to perform an analysis aimed at recovering part of the topo-
graphic energy. The comparison of both values indicates that the improvement
margin is important ηai
ηar
¼ 0.72
0.23 ¼ 3.13 The previous parameters deliver a good measure of the
system’s potential for improvement. However, ηar provides an
even closer examination. It cannot be assumed that perfor-
mance can improve from its actual value (0.23) to the ideal value
(0.72), as the latter does not include losses. J. Water Resour. Plann. Manage. J. Water Resour. Plann. Manage. •
V ¼ 18,580 m3=day (delivered to users). Eti ¼ γ
X
vjðzh −zjÞ ≈γVðzh −zmÞ ¼ 534.62 kWh=day •
V ¼ 18,580 m3=day (delivered to users). =
y
•
V þ ΔV ¼ 19,164 m3=day (supplied) with ΔV ¼ 584 m3=
day. New network with few leaks. •
V þ ΔV ¼ 19,164 m3=day (supplied) with ΔV ¼ 584 m3=
day. New network with few leaks. •
V þ ΔV ¼ 19,164 m3=day (supplied) with ΔV ¼ 584 m3=
day. New network with few leaks. These are very reasonable values and, as shown below,
they have minimal effect on the final estimation. However, it
should be highlighted that if the demand is irregularly dis-
tributed (i.e., concentrated in the lowest nodes), major errors
will result from this simplification. In any case, the necessary
values are often available, and these simplifications are not
needed. y
•
Required pressure by drip emitters (po) = 20 m. •
Consumed
energy:
natural
664 kWh=day,
wire
energy
5,833 kWh=day, total 6,497 kWh=day. •
Consumed
energy:
natural
664 kWh=day,
wire
energy
5,833 kWh=day, total 6,497 kWh=day. =
y
=
y
•
Elevations: zh ¼ 35.53 m; zl ¼ 14.39 m. Natural water level at
suction tank zn ¼ 25 m. =
y
=
y
•
Elevations: zh ¼ 35.53 m; zl ¼ 14.39 m. Natural water level at
suction tank zn ¼ 25 m. Because of the node demands and elevations (vj; zj), the energy
requirements for the ideal system are: If the simplified values had been used (Fig. 8), the results
would
have
been
ηai ¼ 0.74,
θti ¼ 0.26,
ηar ¼ 0.24
and
ηai=ηar ¼ 3.08 (all of these values being quite accurate es-
timations). However, it should be pointed out that the node
synthesizing the global behavior of the network must be
chosen carefully. In this case, because all the data were avail-
able,
the
weighted
average
(node elevation × consumed
volume) was known. J. Water Resour. Plann. Manage. To determine a good,
but realistic performance, a reasonable loss levels must be
established: If the demands and elevation nodes are not available, the values
Euo and Eti can be estimated from the average elevation. If demand
is homogenously distributed, the results are fairly precise. Fig. 8
represents the simplified system for the estimation of Euo and Eti. The simplified expressions for Euo and Eti are The simplified expressions for Euo and Eti are •
Water losses: Their real value is, in this case, very good
at
(584=19,164 ¼ 0.03),
thus
no
estimation
is
required. The current value is used, and the water loss embedded
energy: Euo ≈γ po
γ V þ γVðzm −zlÞ ¼ γV
po
γ þ ðzm −zlÞ
¼ 1,547.44 kWh=day ¼ 1,547.44 kWh=day Fig. 8. Cap de Terme equivalent irrigation network Fig. 8. Cap de Terme equivalent irrigation network © ASCE 04014095-9 04014095-9 J. Water Resour. Plann. Manage. J. Water Resour. Plann. Manage. Erl;o ¼ γ
X
Δvjo
pjr
γ ¼ γ
X
Δvjo
po
γ þ pjt;r
γ
≈γΔVo
po
γ þ
zh −zh þ zl
2
¼ 9,810 · 584
3,600,000
20 þ
35.53 −35.53 þ 14.39
2
¼ 48.61 kWh=day whereas the existing frequency drive motors are set to match the
required pressure (20 m). Irrigation schedules have also been
modified to guarantee that pumps operate at a constant flow rate. Additionally, the system has been decoupled in three zones: high,
medium, and low irrigation areas. Being a tree-like network fed by
four pumps working in parallel, the required cost is assumable. The
result has been to decrease the global θti (must be weighted with the
value of each subnetwork) thus increasing its complementary
value, ηai. The new value, 0.22, is a significant upgrade with regard
to the initial value (0.28). Downloaded from ascelibrary.org by Universidad Politecnica De Valencia on 11/07/14. Copyright ASCE. For personal use only; all rights reserved. •
Friction losses: The assumed average path, Lpm, can be again
estimated with a weighted average (being a tree network, the
length covered by all the consumed volumes is known). In this
case its value is 2 km, with a unit head loss of 2.5 m=km
and 4 m of local losses at the pumping station (where water
is filtered). Then: •
Friction losses: The assumed average path, Lpm, can be again
estimated with a weighted average (being a tree network, the
length covered by all the consumed volumes is known). In this
case its value is 2 km, with a unit head loss of 2.5 m=km
and 4 m of local losses at the pumping station (where water
is filtered). Then: nica De Valencia on 11/07/14. Copyright ASCE. For personal use only; all rights reserved. ¼ 510.46 kWh=day •
Other losses: Ero;o ¼ 0 (none in this network)
In consequence, Esr;o results •
Other losses: Ero;o ¼ 0 (none in this network)
In consequence, Esr;o results Esr;o ¼ ðEuo þ Etr;oÞ þ Erg;o
≈ðEuo þ EtiÞ þ ðErl;o þ Erf;o þ Erp;o þ Ero;oÞ Esr;o ≈ð1,494.23 þ 588.71Þ þ ð48.61 þ 443.88 þ 510.46Þ
¼ 3,085.89 kWh=day Esr;o ≈ð1,494.23 þ 588.71Þ þ ð48.61 þ 443.88 þ 510.46Þ
¼ 3,085.89 kWh=day Acknowledgments The authors acknowledge the very valuable contributions made
by the reviewers of this paper, because their comments and sugges-
tions have helped to significantly improve the contents. Addition-
ally, we thank the staff of Aguas de Valencia for providing helpful
advice and real case studies used to tune the software tool devel-
oped based on this paper. And last but not least, the research
leading to these results received funding from the European
Union Seventh Framework Programme (FP7/2007-2013) under
grant agreement number 265122. The translation of this paper
was funded by the Universitat Politècnica de València, Spain. And the target performance would be ηar;o ¼ Euo
Esr;o
¼ 1,494.23
3,085.89 ¼ 0.48 which is halfway between the real and ideal values (0.23 and
0.72). The conclusion is that there is an important improve-
ment margin (0.48 over 0.23) and efficiency can be doubled. Furthermore, the hypothesis considered to calculate ηar;o does
not affect the final decision: proceeding with an in-depth
analysis (a precise estimation, with an audit, will result in a
final value of 0.48 0.05). Therefore, the authors are in a
position to state that the diagnostic is correct, and that
expectations have been met (Cabrera et al. 2014). Conclusion Improving hydraulic and energy efficiency in PWTS (regardless
of urban or irrigation water use) is an issue that is gaining in
importance—but there are no simple solutions. Before attempting
to find a solution, the implementation of a range of operational (no
investment required, e.g., removing supplied excess energy) and
structural measures (e.g., dividing the system to reduce θti or
installing PATs) is required. Prior to any analysis, a system
diagnostic with the proposed metrics is necessary to estimate with
sufficient precision the real improvement margins. Erf;o ¼ γðV þ ΔVoÞ
LpmJm þ Δprf;p
γ
¼ 9,810 · 19,164
3,600,000
½2 · 2.5 þ 4 ¼ 443.88 kWh=day •
Pumping losses: A (very good) global performance of 0.82 for
the pumping station (variable frequency drive—motor pump) is
selected. Taking into account that the critical point is the furthest
point (Lpc ¼ 2Lpm) shaft losses are This paper presents an assessment methodology that requires
some hypotheses that cannot be rigorously formulated at the initial
stage of the process; and as a consequence, the final ηar;o value
cannot be precisely determined. However, for the purposes of
the diagnostic (based on the order of magnitude of the difference
ηar;o −ηar) this lack of accuracy has no special relevance on the
outcome of the analysis. Furthermore, because this value can be
precisely calculated from an energy audit, an in-depth analysis
of further case studies will help to refine this subjective hypothesis
in the near future. Erp;o ¼ γðV þ ΔVoÞ
LpcJm þ Δprf;p
γ
þ po
γ
þ ðzc −zpÞ
1
ηpo
−1
¼ 9,810 · 19,164
3,600,000
½ð4 · 2.5Þ þ 4 þ 20
þ ð35.53 −25Þ
1
0.82 −1
If the improvement margin (ηar;o −ηar) is deemed relevant, it
should trigger the subsequent stages in the process (not described
in this paper); the first being a water and energy audit to discover
which parts of the system have the greatest improvement margins
and which present the best cost-benefit opportunities. Furthermore,
in systems with an irregular terrain in which topographic energy is
significant, the recovery of this energy should be explored—and if
found to be inviable, then the overpressures should be neutralized
with PRVs. J. Water Resour. Plann. Manage. Notation The following symbols are used in this paper: The following symbols are used in this paper: Actions carried out include the elimination of the initial energy
surplus; one of the five operating pumps is deemed redundant be-
cause the required pressure can be achieved with only four pumps, Ebi = base energy or minimum required energy for the
ideal system (Ebi ¼ Euo þ Eti); y
bi
uo
ti
Eei; Eer = supplied excess energy for the ideal and real systems; Eei; Eer = supplied excess energy for the ideal and real systems; © ASCE © ASCE © ASCE J. Water Resour. Plann. Manage. 04014095-10 J. Water Resour. Plann. Manage. Δt = time period; Erg; Erg;o = reducible global energy (corresponding to real or
target conditions); f;p = local losses in the water pumping station; Erl; Erl;o = reducible energy embedded in leaks (corresponding
to real or target conditions); Δvjo = leaked volume and target leaked volume at a node j
of the system; g
Ero; Ero;o = other reducible energy (corresponding to real or
target conditions); ΔV; ΔVo = total leaked volume and total target leaked volume of
the system; ηai; ηar = ideal and real performance of the system without
recovery; Erp; Erp;o = reducible energy in pumping stations (corresponding
to real or target conditions); g
Eyr = recovered energy (from the topographic energy); ηar;o = target energy efficiency performance of the system
without recovery; y
Hcr; Hhr = piezometric head in the real system at the critical and
highest node; ηpo = target energy efficiency performance for the whole
pumping group; g
Hjr; Hlr = piezometric head in the real system at the generic and
lowest node; ηwi; ηwr = ideal and real performance of the system with energy
recovery; Hhi; H
hi = piezometric head at the highest node (ideal system
without and with pressure excess); θei; θer = percentage excess energy; ideal case ¼ Eei=Esi,
real case ¼ Eer=Esr; Hji; H
ji = piezometric head at the generic node (ideal system
without and with pressure excess); θti; θtr = percentage of total topographic energy;
ideal case ¼ Eti=Esi, real case ¼ Etr=Esr; Hli; H
li = piezometric head at the lowest node (ideal system
without and with pressure excess); λrf = percentage of reducible friction energy related to the
supplied energy (Erf=Esr); hpi; h
pi = head pump in the ideal system (without and with
excess pressure); f
λrg = percentage of reducible global energy related to the
supplied energy (Erg=Esr); hpr = head pump in the real system; g
λrl = percentage of reducible energy embedded leaks
related to the injected energy (Erl=Esr); p
hni; hnr = natural supplied head in ideal and real systems
respectively (usually hni ¼ hnr); Iei; Ier = ideal and real energy intensity, considering all
supplied energy (natural and wire); λro = percentage of other energy losses related to the
supplied energy (Ero=Esr); and I 0er = real energy intensity using only wire energy; λrp = percentage of reducible energy in pumping related to
the supplied energy (Erp=Esr). J. Water Resour. Plann. Manage. Esi; Esr = total supplied energy for the ideal and real systems,
respectively; po = required pressure (established by
standards); Esr;o = target energy to be supplied to the system; qk = average flow in pipe k; Eti; Etr = topographic energy required by the ideal and real
system, respectively; V = total volume demanded by the system; vj = volume demand at node j; Etr;o = topographic energy of the system working with the
target conditions; zc; zh; zj; zl = critical (c), highest (h), generic (j) and
lowest node (l) elevation, respectively; Euo = minimum required energy by users (constant, no
matter the system be real or ideal); zm = average of the extreme nodes elevation; zm = average of the extreme nodes elevation; zn = reservoir or tank water natural level elevation
(= pump suction level); zn = reservoir or tank water natural level elevation
(= pump suction level); Erf; Erf;o = reducible friction energy (corresponding to real or
target conditions); γ = water specific weight (N=m3); γ = water specific weight (N=m3); γ = water specific weight (N=m3); nica De Valencia on 11/07/14. Copyright ASCE. For personal use only; all rights reserved. J. Water Resour. Plann. Manage. J. Water Resour. Plann. Manage. Jm = average pipe head loss (m=km); Lpc = distance between the pumping station and the critical
point; Subscripts Lpm = average distance between the pumping station and
the consumption nodes; a = achievable; Pmin = fixed power value to select candidate pipes for
placing PATs stations; b = base or minimum; pci; p
ci = pressure at the critical node (ideal system without
and with excess pressure); pcr; pcr = pressure at the critical node (real system without and
with excess pressure); pei; per = excess pressure of the system (ideal and real case); phi; p
hi = pressure at the highest node (ideal system without
and with excess pressure); phr; pjr; plr = pressure in the real system at the highest, generic
and lowest node, respectively; pht;r = topographic real pressure at highest node (pht;r ¼ 0
if critical node = highest node); pji; p
ji = pressure at the generic node (ideal system without
and with excess pressure); pjt;i; pjt;r = topographic pressure at generic node (ideal and real
systems); pli; p
li = pressure at the lowest node (ideal system without and
with excess pressure); plt;i; plt;r = topographic pressure at lowest node (ideal and real
systems); © ASCE © ASCE 04014095-11 J. Water Resour. Plann. Manage. pr
Δvj; Δ References Lingireddy, S., and Wood, D. (1998). “Improved operation of water
distribution systems using variable-speed pumps.” J. Energy Eng.,
10.1061/(ASCE)0733-9402(1998)124:3(90), 90–103. Cabrera, E., Cabrera, E., Jr., Cobacho, R., and Soriano, J. (2014). “Towards
an energy labeling of pressurized water systems.” Procedia Eng., 70,
209–217. Official Journal of the European Union (OJEU). (2012). “Directive 2009/
125/EC of the European parliament and of the council with regard to
ecodesign requirements for water pumps.” European Commission,
Brussels, Belgium. Cabrera, E., Pardo, M. A., Cobacho, R., and Cabrera, E., Jr. (2010). “Energy audit of water networks.” J. Water Resour. Plann. Manage.,
10.1061/(ASCE)WR.1943-5452.0000077, 669–677. Carravetta, A., Fecarotta, O., Del Giudice, G., and Ramos, H. M. (2012). “Energy production in water distribution networks: A PAT design
strategy.” J. Water Resour. Plann. Manage., 26(13), 3947–3959. Ormsbee, L. E., Walski, T. M., Chase, D. V., and Sharp, W. W. (1989). “Methodology for improving pump operation efficiency.” J. Water
Resour. Plann. Manage., 10.1061/(ASCE)0733-9496(1989)115:2(148),
148–164. Carravetta, A., Fecarotta, O., Sinagra, M., and Tucciarelli, T. (2014). “Cost-benefit analysis for hydropower production in water distribution
networks by a pump as turbine.” J. Water Resour. Plann. Manage., 10
.1061/(ASCE)WR.1943-5452.0000384, 04014002. Papa, F., Radulj, D., Karney, B., and Robertson, M. (2013). “Pump energy
efficiency field testing & benchmarking in Canada.” Proc., Int. Conf. on
Asset Management for Enhancing Energy Efficiency in Water and
Wastewater Systems, IWA. Marbella, Spain. Carriço, N., Covas, D., Alegre, H., and Almeida, M. (2013). “How to assess
the effectiveness of energy management processes in water supply sys-
tems.” Proc., Int. Conf. on Asset Management for Enhancing Energy
Efficiency in Water and Wastewater Systems, IWA. Marbella, Spain. Pelli, T., and Hitz, H. U. (2000). “Energy indicators and savings in water
supply.” AWWA J., 92(6), 55–62. Perez-Urrestarazu, L., and Burt, C. M. (2012). “Characterization of pumps
for irrigation in central California: Potential energy savings.” J. Irrig. Drain. Eng., 10.1061/(ASCE)IR.1943-4774.0000463, 815–822. Colombo, A., and Karney, B. (2002). “Energy and costs of leaky pipes:
Toward comprehensive picture.” J. Water Resour. Plann. Manage.,
10.1061/(ASCE)0733-9496(2002)128:6(441), 441–450. Rossman, L. A. (2000). “EPANET 2: User’s manual.” U.S. Environmental
Protection Agency, Cincinnati. Duarte, P., Covas, D. I. C., and Alegre, H. (2009). “PI for assessing effec-
tiveness of energy management processes in water supply systems.” Int. Conf. PI09: Benchmarking water services—The way forward, IWA,
Amsterdam, Netherlands. Ulanicki, B., Kahler, J., and See, H. (2007). “Dynamic optimization ap-
proach for solving an optimal scheduling problem in water distribution
systems.” J. Superscripts Giustolisi, O., Savic, D., and Kapelan, Z. (2008). “Pressure-driven demand
and leakage simulation for water distribution networks.” J. Hydraul. Eng., 10.1061/(ASCE)0733-9429(2008)134:5(626), 626–635. * = indicates excess of pressure (more than the required
value po); and * = indicates excess of pressure (more than the required
value po); and Kurek, W., and Ostfeld, A. (2014). “Multiobjective water distribution
systems control of pumping cost, water quality, and storage-reliability
constraints.” J. Water Resour. Plann. Manage., 10.1061/(ASCE)WR
.1943-5452.0000309, 184–193. ′ = indicates that the energy intensity indicator, I 0er only
includes wire energy. Lambert, A., Brown, T. G., Takizawa, M., and Weimer, D. (1999). “A re-
view of performance indicators for real losses from water supply
systems.” J. Water SRT: Aqua, 48, 227–237. J. Water Resour. Plann. Manage. References Water Resour. Plann. Manage., 10.1061/(ASCE)0733
-9496(2007)133:1(23), 23–32. Fontana, N., Giugni, M., and Portolano, D. (2012). “Losses reduction and
energy production in water-distribution networks.” J. Water Resour. Plann. Manage., 10.1061/(ASCE)WR.1943-5452.0000179, 237 –244. Walski, T. M. (1993). “Tips for saving energy in pumping operations.” J. Am. Water Works Assoc., 85(7), 49–53. Giustolisi, O., Laucelli, D., and Berardi, L. (2013). “Operational optimiza-
tion: Water losses versus energy costs.” J. Hydraul. Eng., 10.1061/
(ASCE)HY.1943-7900.0000681, 410–423. Water in the West (WW). (2013). “Water and energy nexus: A literature
review.” Stanford Univ., Standford, CA. © ASCE 04014095-12 J. Water Resour. Plann. Manage. J. Water Resour. Plann. Manage. J. Water Resour. Plann. Manage.
|
https://openalex.org/W2000604300
|
http://www.scielo.br/pdf/ci/v30n3/7283.pdf
|
Portuguese
| null |
Integração e interoperabilidade no acesso a recursos informacionais eletrônicos em C&T: a proposta da Biblioteca Digital Brasileira
|
Ciência da Informação
| 2,001
|
cc-by-sa
| 7,061
|
Palavras-chave Bibliotecas digitais; Publicações eletrônicas; Arquivos abertos;
Interoperabilidade; Metadados; Padrões; Tecnologia da informação;
Informação em ciência e tecnologia; Comunicação científica; Acesso à
informação. Abstract This paper describes technological and methodological options to achieve
interoperability in accessing electronic information resources, available in
Internet, in the scope of Brazilian Digital Library in Science and
Technology Project - BDL, developed by Brazilian Institute for Scientific
and Technical Information - IBICT. It stresses the impact of the Web on
publishing and communication in science and technology and also on
information systems and libraries. The work stresses the two main
objectives of BDL project: promoting electronic publishing of different full
text materials - theses, journal articles, papers in events, “grey” literature -
by Brazilian scientific community, so amplifying their nationally and
internationally visibility; and achieving, through a gateway, interoperability
among those heterogeneous electronic information resources available in
the Web, thus avoiding a user to navigate and query those resources
one by one separately. Integration and interoperability in accessing
electronic information resources in science and
technology: the proposal of Brazilian Digital
Library Do ponto de vista da informação como subsídio às
atividades acadêmicas e em C&T, a Internet vem
proporcionar facilidades que extrapolam o conceito
tradicional de informação bibliográfica baseada em
documentos, como artigos de periódico, trabalhos em
congressos, teses etc. Novos recursos informacionais estão
à disposição da comunidade de pesquisa além desses
tradicionais, agora em versão eletrônica, como documentos
multimídia, listas de discussão, fóruns eletrônicos,
conferências em linha, imagens (de satélites, de
microscópios, em tempo real), modelos animados, bancos
de preprints eletrônicos, os e-prints etc. Estes recursos tanto
servem de subsídio à pesquisa quanto de canais de
comunicação e publicação dos resultados e de garantia de
primado e originalidade intelectuais dos mesmos. INTRODUÇÃO Carlos Henrique Marcondes
Doutor em ciência da informação, DEP-IBICT/UFRJ. Consultor do
projeto BDB/IBICT. Universidade Federal Fluminense,
Departamento de Ciência da Informação. marcondes@alternex.com.br. Carlos Henrique Marcondes
Doutor em ciência da informação, DEP-IBICT/UFRJ. Consultor do
projeto BDB/IBICT. Universidade Federal Fluminense,
Departamento de Ciência da Informação. marcondes@alternex.com.br. “Um laboratório sem uma biblioteca é como se fosse um animal
descorticado: as atividades motoras continuam a funcionar, mas
falta a coordenação da memória e da vontade” (Zilman, 1979,
p. 115) Resumo Descreve as opções tecnológicas e metodológicas para atingir a
interoperabilidade no acesso a recursos informacionais eletrônicos,
disponíveis na Internet, no âmbito do projeto da Biblioteca Digital
Brasileira em Ciência e Tecnologia, desenvolvido pelo Instituto Brasileiro
de Informação em Ciência e Tecnologia (IBICT). Destaca o impacto da
Internet sobre as formas de publicação e comunicação em C&T e sobre os
sistemas de informação e bibliotecas. São explicitados os objetivos do
projeto da BDB de fomentar mecanismos de publicação pela comunidade
brasileira de C&T, de textos completos diretamente na Internet, sob a
forma teses, artigos de periódicos, trabalhos em congressos, literatura
“cinzenta”, ampliando sua visibilidade e acessibilidade nacional e
internacional, e também de possibilitar a interoperabilidade entre estes
recursos informacionais brasileiros em C&T, heterogêneos e distribuídos,
através de acesso unificado via um portal, sem a necessidade de o
usuário navegar e consultar cada recurso individualmente. A convergência e o uso integrado das tecnologias de
comunicação, de computação e de conteúdos em formato
digital, cujo paradigma é a Internet, tem contribuído nos
anos recentes para criar um novo ambiente de acesso,
disseminação, cooperação e promoção do conhecimento
em uma escala global. Estamos em meio a este processo,
cujas conseqüências ainda não podemos avaliar
completamente. Novos suportes de conhecimento, que
não guardam similares com os materiais impressos em
papel, estão sendo inventados a cada dia. Luís Fernando Sayão Doutor em ciência da informação, DEP-IBICT/UFRJ. Consultor do
projeto BDB/IBICT. Comissão Nacional de Energia Nuclear. Centro de Informações Nucleares
lsayao@cnen.gov.br Há muito tempo está constatada a relação entre
desenvolvimento econômico e social e o estágio de
desenvolvimento da ciência e tecnologia de um país. À ciência e tecnologia está reservado um papel
fundamental na luta pelo desenvolvimento da sociedade
brasileira. Informação é insumo fundamental para o
desenvolvimento da ciência. É em torno do problema da
otimização dos fluxos e da transferência de informação
científica que surge, na segunda metade do século XX, a
ciência da informação (Pinheiro, 1995). Há muito tempo está constatada a relação entre
desenvolvimento econômico e social e o estágio de
desenvolvimento da ciência e tecnologia de um país. À ciência e tecnologia está reservado um papel
fundamental na luta pelo desenvolvimento da sociedade
brasileira. Informação é insumo fundamental para o
desenvolvimento da ciência. É em torno do problema da
otimização dos fluxos e da transferência de informação
científica que surge, na segunda metade do século XX, a
ciência da informação (Pinheiro, 1995). Ci. Inf., Brasília, v. 30, n. 3, p. 24-33, set./dez. 2001 Keywords última década, permitiram o surgimento de um novo
patamar para esses sistemas: antes orientados basicamente
para recuperação de referências bibliográficas em bases de
dados isoladas e textos em papel, voltam-se hoje para a
recuperação distribuída de objetos digitais – textos
completos, imagens em movimento, som etc. –,
estabelecendo como palavras de ordem a publicação na
Internet e a interoperabilidade entre fontes de informação
heterogêneas e globalmente distribuídas. Com o projeto da Biblioteca Digital Brasileira, o IBICT
quer abrir a possibilidade, fomentar e fornecer meios para
que a comunidade brasileira de C&T possa publicar seus
trabalhos de forma rotineira, diretamente na rede, aumentando
com isso sua visibilidade nacional e internacional,
otimizando o fluxo da comunicação científica e reduzindo
o ciclo de geração de novos conhecimentos. Por outro lado, somente a disponibilidade de textos
brasileiros em C&T on-line não teria grande impacto sobre
a comunicação científica e a ciência no país sem a
existência de serviços de informação que viabilizem o
acesso de forma fácil a estes conteúdos. O país também
tem acumulado experiências bastante significativas,
embora isoladas, na criação de bibliotecas digitais e
repositórios de informações na rede. À medida que
experiências brasileiras neste sentido se multiplicam, como
o Prossiga, o Scielo, o repositório de teses da USP, o arquivo
de e-prints do Impa (http://www.preprint.impa.br/
indexEngl.html) etc., disponibilizando de forma crescente
recursos informacionais em texto completo na Web, fica
patente, para as organizações brasileiras que trabalham
com sistemas de informação para C&T, a importância da
questão da interoperabilidade entre bibliotecas digitais e
outros recursos informacionais digitais: como consultar, de
uma única vez, todas estas fontes de forma integrada e
transparente, com o mínimo de esforço, com a máxima rapidez,
e obter resultados consolidados? Dessa forma, os paradigmas de comunicação científica,
tendo por base o periódico científico em papel, com seu
esquema de revisão por pares e o monopólio das grande
editoras científicas, vêm sofrendo grande impacto com o
surgimento da Internet e grande questionamento por parte
da comunidade científica de todo o mundo. Desde o
surgimento do primeiro arquivo eletrônico de preprints,
ou eprints, o ArXiv, no Los Alamos National Laboratory,
criado em 1991 pelo físico Paul Ginsparg (Ginsparg, 1996),
que a própria comunidade científica internacional oferece
uma alternativa prática para a publicação de seus
trabalhos. Uma lista que permite dimensionar a amplitude
dos arquivos eletrônicos por todo o mundo pode ser
encontrada em http://www.osti.gov/eprints/ppnbrowse. Keywords Digital libraries; Electronic publishing; Open archives; Interoperability;
Metadata; Standards; Information technology; Science and technology
information; Scientific communication; Information access. Ci. Inf., Brasília, v. 30, n. 3, p. 24-33, set./dez. 2001 24 Integração e interoperabilidade no acesso a recursos informacionais eletrônicos em C&T: a proposta da ... Integração e interoperabilidade no acesso a recursos informacionais eletrônicos em C&T: a proposta da ... Mais que somente recursos informacionais, os novos
recursos disponíveis via Internet são acima de tudo novas
ferramentas cognitivas, no sentido emprestado a elas por
Pierre Lévy (1993), capazes de abrir novas possibilidades
cognitivas e intelectuais que extrapolam em muito aquelas
oferecidas por documentos em papel, de leitura linear. Para
muitos autores, a Internet representa, neste sentido, uma
mudança de paradigma comparável à invenção da
imprensa por Gutemberg. Esta mudança de paradigma se
faz sentir também no aspecto da comunicação científica. Mais que somente recursos informacionais, os novos
recursos disponíveis via Internet são acima de tudo novas
ferramentas cognitivas, no sentido emprestado a elas por
Pierre Lévy (1993), capazes de abrir novas possibilidades
cognitivas e intelectuais que extrapolam em muito aquelas
oferecidas por documentos em papel, de leitura linear. Para
muitos autores, a Internet representa, neste sentido, uma
mudança de paradigma comparável à invenção da
imprensa por Gutemberg. Esta mudança de paradigma se
faz sentir também no aspecto da comunicação científica. A Internet é um mecanismo de comunicação de alcance
mundial, instantâneo, interativo e multidirecional:
qualquer um pode publicar nela, o que foi publicado é
imediatamente acessível, o autor pode receber um retorno
e avaliação imediatos sobre o que publicou, de qualquer
lugar. Um autor acadêmico almeja a máxima divulgação
para seus trabalhos, para que os resultados de sua pesquisa
tenham o maior impacto possível sobre as pesquisas de
seus pares e sobre outras publicações. Estudos recentes
confirmam que as publicações eletrônicas são muito mais
citadas que as publicações em papel: “The mean number of
citations to offline articles is 2,74, and the mean number of
citations of online articles is 7.03, an increase of 157%”
(Lawrence). Desenvolver mecanismos de publicação
eletrônica para a comunidade acadêmica brasileira,
aumentando sua visibilidade, torna-se, portanto, uma
questão essencial para o desenvolvimento e maturidade
da pesquisa científica brasileira. O PROBLEMA DA INTEROPERABILIDADE os localizadores de informações especializados, como o
GILS (http://www.usgs.gov/gils/) ou portais temáticos
como o SIGNPOST (http://www.signpost.org) americano,
o OMNI (http://www.omni.ac.uk) e o SOSIG (http://
www.sosig.ac.uk)ingleses, o PROSSIGA – Comunicação
e Informação para a Pesquisa – (http://www.prossiga.br)
ou LIS – Localizador de informações em Saúde – (http://
www.bireme.br) no Brasil. Um aspecto problemático da cultura de nosso tempo é o
assim chamado fenômeno da explosão informacional, a
grande quantidade de informações produzidas e
disponibilizadas por diferentes atividades sociais,
dificultando sua identificação, acesso e utilização. Na
emergência da sociedade da informação, o valor desta como
insumo para qualquer atividade, seja ela uma decisão
econômica, um processo cultural ou de ensino/
aprendizagem, uma pesquisa científica ou tecnológica, está
relacionado diretamente ao seu potencial de orientar de
forma econômica o dispêndio de energia para a realização
desta atividade. Para que possa realizar todo este potencial,
a informação relevante para um dado problema deve estar
disponível no tempo certo. De nada adianta a informação
existir, se quem dela necessita não sabe da sua existência
ou se ela não puder ser encontrada. Ambas as alternativas, os mecanismos de busca gerais e os
portais temáticos oferecem soluções parciais para a
localização de informações na Internet, principalmente as
de interesse para C&T. As deficiências dos mecanismos de
busca são já bastante conhecidas e discutidas na literatura
(Sneiderman,1997). Entre as principais, pode-se citar as
seguintes: baixa qualidade da indexação, por ser feita
automaticamente, que resulta em grande quantidade de
informações recuperadas, a maioria sem relevância (em
termos de recuperação de informação, oferecem alta
revocação, mas baixa precisão); cobertura parcial da
Internet; as ferramentas de busca não são especializadas;
indexam páginas HTML isoladas, e não recursos; além
disto, grande quantidade de informações disponíveis na
Internet estão sob a forma de registros contidos em bases
de dados, que ficam assim “escondidas”; estes registros
são acessados somente por meio das interfaces destas bases
de dados, o que pressupõe uma interação entre um usuário
humano com a base de dados e, portanto, ficam inacessíveis
aos programas robôs. Esta situação assume proporções alarmantes com o
surgimento da Internet. Uma notícia divulgada no Boletim
Edupage em português, de 05/04/98, publicado pela RNP,
levanta o problema da busca de informações na Internet e
comenta os resultados de um estudo sobre o desempenho
dos assim chamados “mecanismos de busca”: “ACHANDO UMA AGULHA (OU 7.079 PÁGINAS
EM UMA AGULHA) NA WEB Keywords html A alternativa dos e-prints vem também se articulando
na chamada OpenArchives Initiative (http://www. openarchivesinitiative.org) São assim dois os objetivos fundamentais do projeto BDB:
a) fomentar a publicação de recursos informacionais de interesse
para C&T na rede, propiciando à comunidade científica
brasileira meios para publicar diretamente na Web, dando
maior visibilidade à produção brasileira em C&T, tanto
nacionalmente, quanto internacionalmente; b) viabilizar
o acesso rápido e integrado a estes recursos, facilitando a
descoberta na Internet de recursos informacionais
brasileiros de interesse para a ciência e tecnologia, de
forma integrada e, dessa forma, encurtando o ciclo de
comunicação científica entre pares nas comunidades
brasileiras de C&T. Estas transformações têm exercido profunda influência
sobre a concepção e funcionamento dos sistemas de
informação automatizados, especialmente aqueles
voltados para as atividades de pesquisa. O rompimento de
barreiras tecnológicas importantes, experimentadas na Ci. Inf., Brasília, v. 30, n. 3, p. 24-33, set./dez. 2001 25 Carlos Henrique Marcondes / Luís Fernando Sayão “ACHANDO UMA AGULHA (OU 7.079 PÁGINAS
EM UMA AGULHA) NA WEB Como desvantagens este esquema apresenta:
manutenção pelo provedor de dados da base comum de
metadados; grande ônus administrativo e gerencial por
parte do provedor de serviços para sincronizar o envio
dos dados por parte dos provedores de dados e processá-
los para incluí-los na base comum de metadados;
necessidade de sincronização entre os dados armazenados
nos provedores de dados e os metadados coletados pelo
provedor de serviços. Hoje, no cenário mundial, identificam-se várias
alternativas de interoperabilidade e acesso integrado a
recursos informacionais heterogêneos publicados na rede. Estas podem ser agrupadas basicamente em duas
alternativas, embora ainda não tenha se fixado uma
nomenclatura amplamente aceita: buscas distribuída a
diferentes servidores e busca em uma base de metadados
centralizada. Em ambas as alternativas, o usuário interage
com uma única interface Web, de onde é submetida a busca. Na primeira alternativa, a interface de busca distribui a
consulta (broadcast search) a diferentes sites, segundo um
protocolo padrão, identificados pela interface como
capazes de fornecer respostas satisfatórias, e os resultados
são consolidados e integrados. Exemplo típico desta
alternativa é o conhecido protocolo Z39.50, usado para
proporcionar interoperabilidade entre catálogos
automatizados de bibliotecas. Esta alternativa apresenta
as seguintes vantagens: novos provedores de dados podem
ser acrescentados, desde que sejam aderentes ao padrão ou
padrões utilizados, com reconfiguração mínima. Como
desvantagens, pode-se apontar que provedores de dados
precisam rodar software servidor do protocolo padrão para
serem consultáveis. Alguns destes softwares consomem
muitos recursos por parte dos provedores de dados, como
no caso do Z39.50 (Troll, 2001). Em alguns casos, são
necessários servidores especializados, como os servidores
de índices, que roteiam as consultas para os servidores
capazes de atendê-las. Alguns dos padrões tecnológicos
utilizados são os seguintes: Z39.50 (ISO/NISO), O esquema de coleta automática de metadados (harvesting)
é mais recente: metadados de diversos provedores de
informação tornam-se ¨visíveis¨ através de protocolos
padronizados e são coletados automaticamente de forma
periódica e armazenado em um data warehousing, ou base
centralizada de metadados, onde são efetuadas as buscas
de forma integrada. Vantagens deste esquema são as
seguintes: novos provedores de dados podem ser
acrescentados, desde que sejam aderentes ao padrão
utilizado; melhor desempenho em função de a consulta
ser na base de metadados local do provedor de serviços. Desvantagens: manutenção pelo provedor de dados da
base comum de metadados; necessidade de sincronização
entre os dados armazenados nos provedores de dados e os
metadados coletados pelo provedor de serviços (Liu,
2001). “ACHANDO UMA AGULHA (OU 7.079 PÁGINAS
EM UMA AGULHA) NA WEB Integração e interoperabilidade no acesso a recursos informacionais eletrônicos em C&T: a proposta da Whois++, LDAP, CIP, SDLIP, DIENST. Esta alternativa
é utilizada nos seguintes sistemas: NCSTRL (University
of. Cornell, EUA), NDLTD – Networked Digital Library
of Theses and Dissertations - federated search (Powel,
1998), California Digital Library (http://www.cdlib.org/),
Berkeley Environmental Digital Library, EUA, ROADS,
ISAAC/SCOUT Project (University of Stanford, EUA). A questão começa a ser levantada na An Intenational
research agenda for digital libraries, de 1998 – um agenda de
pesquisa conjunta da NSF (EUA) e União Européia – em
três grupos de trabalho temáticos: global resource discovery,
interoperability e metadata. Hoje é endereçada diretamente
por diferentes iniciativas de pesquisa, como a Joint NSF –
JISC International Digital Libraries Research Programme,
como o consórcio Imesh (http://www.desire.org/html/
subjectgateways/community/imesh/), o Scout Project, e
por iniciativas práticas como OpenArchives Initiative,
Arc - Cross Archive Searching Service, NCSTRL (Univ. Cornell, EUA), NDLTD (Virginia Tech, University,
EUA), Digital Library Federation (consórcio de
bibliotecas digitais americanas), ROADS (UKOLN JISC,
Inglaterra). As diferentes denominações sob as quais o
tema aparece na literatura convergem para os conceitos
de integração e interoperabilidade entre bibliotecas
digitais, que consistiria na possibilidade de um usuário
realizar buscas a recursos informacionais heterogêneos,
armazenados em diferentes servidores na rede, utilizando-se de
uma interface única sem tomar conhecimento de onde nem como
estes recursos estão armazenados. Na segunda alternativa, metadados referentes a
documentos eletrônicos são coletados periodicamente,
alimentando uma base comum de metadados sobre a qual
são realizadas as buscas. Este esquema é bastante
conhecido da colaboração/cooperação entre as instituições
participantes para manutenção do Catálogo Coletivo/base
de metadados centralizada. Dentro desta alternativa
variam os esquemas de centralização destes metadados. O esquema do envio de metadados por parte das
instituições cooperantes é mais tradicional e largamente
conhecido pela comunidade de informação, inclusive a
brasileira, em sistemas/bases de dados como LILACS/
BIREME, SITE/IBICT, INIS/CIN. Este esquema apresenta
as seguintes vantagens: novos provedores de dados podem
ser acrescentados, desde que sejam aderentes ao padrão ou
padrões utilizados; melhor desempenho em função da
consulta ser na base de metadados local do provedor de
serviços. “ACHANDO UMA AGULHA (OU 7.079 PÁGINAS
EM UMA AGULHA) NA WEB Um estudo realizado pelo NEC Research Institute afirma que a
Internet explodiu para mais de 320 milhões de páginas na Web,
uma estimativa que não inclui milhões de páginas com acesso
protegidas por senhas ou “muros de pesquisa” que bloqueiam
acesso a browsers ou mecanismos de busca. O estudo indica
que a pesquisa do mecanismo de busca HotBot tem o índice
mais abrangente da Web, mas, ainda assim, cobre apenas 34%
das páginas indexáveis. A cobertura de alguns dos outros
mecanismos de busca inclui: AltaVista (28%); Northern Light
(20%); Excite (14%); Lycos (3%).” Por sua vez, as bibliotecas digitais e os portais temáticos
isolados resolvem somente em parte o problema do acesso
a recursos informacionais de interesse para C&T publicados
na rede: continuam limitadas ao “seu” acervo. Descobrir,
avaliar, tratar e indexar estes recursos por profissionais de
informação é caro e lento. Estes recursos estão sendo criados
em número crescente, armazenados em diferentes
servidores isolados, operados por interfaces de busca
diferentes, o que obriga um usuário a uma dispendiosa busca,
site a site, para encontrar informações relevantes. Uma novidade em termos de mecanismos de busca que
parece alentadora são os projetos CLEVER e GOOGLE
(http://www.google.com), com suas propostas de
ordenamento e priorização (ranking) dos resultados de uma
busca, tendo por base os sites mais referenciados por links a
partir de outros (Clever, 1999). Do ponto de vista de um usuário acadêmico ou pesquisador,
o interessante e confortável seria poder submeter sua
necessidade de informação e interagir com uma única
interface e ter retornadas informações de diferentes fontes,
de forma consolidada. Este é um tema que, sob diferentes
denominações, está sendo cada vez mais discutido: digital
libraries federation e distributed archives (Liu, 2001),
confederated digital libraries (Leiner, 1998), distributed subject
gateways (IMesh, 1999), networked digital library (Davis,
1995), multiple information sources (Paepcke, 2000) cross-
searching, heterogeneous distributed databases, metasearches
(Gravano, 1996) etc. A enorme quantidade de informação armazenada e
disponibilizada via Internet torna cada vez mais crítico o
problema da identificação de informação relevante, assim
chamada information discovery. Diferentes estratégias para
fazer frente à explosão informacional trazida pela Internet
podem hoje ser divisadas, como os mecanismos de busca
gerais (AltaVista, Excite, Lycos, Infoseek, Yahoo e outros), Ci. Inf., Brasília, v. 30, n. 3, p. 24-33, set./dez. 2001 26 ção e interoperabilidade no acesso a recursos informacionais eletrônicos em C&T: a proposta da ... MODELO DE INTEROPERABILIDADE
DA BDB O projeto de implantação da BDB no país
pressupõe forte ação de integração, liderada pelo
IBICT, dos mais importantes provedores de
conteúdos e de serviços de informação para
C&T do país. Esta integração se dará em torno
das questões prioritárias para a BDB, que são a
publicação e a disponibilidade de textos
completos e outros objetos digitais na Internet
e a interoperabilidade entre os diversos
sistemas/serviços de informação participantes
através de um portal único de acesso,
preservando-se
a
independência
e
peculiaridades de cada sistema/serviço
participante. Para conseguir cooperação dos
eventuais provedores de dados, o conjunto de
metadados, a configuração, os padrões e procedimentos
por parte dos provedores de dados para garantir
interoperabilidade com a BDB deverão ser os mais simples
e menos onerosos para os provedores de dados, garantindo
sua máxima independência. No intuito de disseminar mais facilmente as tecnologias
de publicação na Internet entre as diversas comunidades
de conhecimento, sempre que possível serão adotados
software de domínio público, preservada a qualidade,
documentação e mantenabilidade dos mesmos. Isto se deve
à constatação de que há uma diversidade surpreendente
de software livres, confiáveis e de qualidade que estão sendo
adotados por instituições importantes na área de
informação. Apesar do avanço acelerado das tecnologias Web e das
tecnologias de informação e comunicação, o projeto da
BDB prevê a utilização de tecnologias consolidadas, cujo
grau de estabilidade e confiabilidade tenham sido
comprovados no país e no exterior, e que, prioritariamente,
tenham sido aplicados nas principais experiências
internacionais análogas à da proposta da BDB. Além do
mais, essas tecnologias devem ser passíveis de serem
implantadas, mantidas e, quando necessário, alteradas pelo
corpo técnico do IBICT. Sempre que possível, serão
adotadas tecnologias abertas, não proprietárias, que
permitam garantir o grau de interoperabilidade desejável
entre os diversos partícipes da BDB e que possam
facilmente ser repassadas aos parceiros do Projeto. Esta
opção tem como objetivo a disseminação no país de um
corpo de protocolos e padrões que possam ser adotados
pelos futuros integrantes da BDB e por outros sistemas
que queiram aderir a sistemas/redes internacionais. O modelo de interoperabilidade proposto para a BDB
aproxima-se bastante dos modelos do portal da NDLTD
(Suleman, 2001) e do Arc – Cross Archive Searching
Service (http://www.arc.cs.odu.edu). Ambos os sistemas
fazem harvesting de metadados de provedores de dados,
alimentando uma base de dados central de metadados. O portal da BDB na Internet será a materialização da
Biblioteca Digital Brasileira em C&T. FIGURA 1 (Virginia Technical University), do portal da
NDLTD (Suleman, 2001), da OpenArchives
Initiative, que, através do protocolo OAI
harvest protocol, permite colheita (harvesting)
de metadados, do Arc – Cross Archive
Searching Service, primeiro serviço a
proporcionar acesso integrado a diversos
arquivos
eletrônicos
(http://
www.arc.cs.odu.edu). “ACHANDO UMA AGULHA (OU 7.079 PÁGINAS
EM UMA AGULHA) NA WEB Padrões utilizados: OpenArchives Harvest
Protocol, Open Archives Metadata Set, Dublin Core,
XML. Exemplos são as experiências do sistema MARIAN 27 Ci. Inf., Brasília, v. 30, n. 3, p. 24-33, set./dez. 2001 Carlos Henrique Marcondes / Luís Fernando Sayão FIGURA 1
Proposta de interface de busca heterogênea para a BDB FIGURA 1
Proposta de interface de busca heterogênea para a BDB MODELO DE INTEROPERABILIDADE
DA BDB Trata-se de um site
que, através de diferentes mecanismos de interopera-
bilidade, possibilitará ao pesquisador acesso unificado e
integrado a diferentes recursos infor-macionais brasileiros
de interesse para C&T, heterogêneos e distribuídos, sem a
necessidade de navegar e consultar cada recurso
individualmente. A figura 1 dá uma idéia da proposta do
portal da BDB. Ci. Inf., Brasília, v. 30, n. 3, p. 24-33, set./dez. 2001 28 Integração e interoperabilidade no acesso a recursos informacionais eletrônicos em C&T: a proposta da ... Entre estes recursos informacionais se
incluirão, já na versão inicial do portal,
periódicos eletrônicos brasileiros que fazem
parte do portal Scielo, mantido pela
Bireme, anais eletrônicos de eventos
brasileiros em C&T a serem disponi-
bilizados pela CNEN/CIN, bancos de teses
eletrônicas hoje já existentes na USP,
Unicamp, UFSC, PUC-Rio, ENSP/Fiocruz,
repositórios de e-prints brasileiros que
começam a ser dissemindos na Internet,
como o do Impa. Na figura 2, é mostrado o
modelo geral de interoperabilidade da BDB. FIGURA 2 FIGURA 2 FIGURA 2 Em um ambiente de publicação e acesso a
documentos publicados na rede como o da
BDB, a OpenArchives Initiative reconhece
dois atores institucionais fundamentais:
provedores de dados e provedores de
serviço. Estas definições serão utilizadas aqui para
explicitar a solução de interoperabilidade adotada, com
base no que é estabelecido na OpenArchives Initiative: FIGURA 5 elemento (Dublin Core, 2000) e pode ser codificado em
formatos como HTML e XML (Cox, 2000), (Beckett,
2000), (Beckett. 2001). Como é explicitado na própria
proposta Dublin Core, o conjunto de metadados deve ser
tão simples e intuitivo a ponto de permitir que o próprio
autor descreva seu trabalho. É exatamente assim que
funcionam os ambientes de submissão de trabalhos: ao
submeter seu trabalho, o autor preenche um formulário
com os metadados pertinentes. Estratégia de interoperabilidade
[COLETA AUTOMÁTICA DE METADADOS] FIGURA 4
Solução Técnica
[PROCESSOS DE HARVESTING] Além de integrar, via consultas
distribuídas, os recursos informacionais
servidos pelo protocolo Z39.50, a BDB
manterá em seu site uma base comum de
metadados, obtida pelo processo de
harvesting dos metadados dos recursos/
serviços de informação que não tiverem
servidor Z39.50. Estes recursos/serviços
serão objeto de coleta automática
periódica, usando o OAI Harvesting
protocol dos provedores de dados
compatíveis com este protocolo. Outras
soluções foram pensadas de modo a não
onerar tecnicamente provedores de
dados não compatíveis com este
protocolo, como coleta de metadados via
FTP em arquivos HTML ou arquivos
texto. Os formatos de arquivos passíveis
de coleta automática são ilustrados na
figura 4. Estes metadados serão processados e
armazenados na base comum de metadados, mantida no
site da BDB; esta base, por sua vez, contará também com
um servidor protocolo Z39.50, que a tornará acessível a
partir do portal da BDB como qualquer outro recurso
servido por este protocolo. Arquivos acadêmicos de
preprints eletrônicos que poderão ser implantados em
departamentos de instituições de ensino superior,
institutos de pesquisa, sociedades científicas, periódicos
eletrônicos, como o próprio Ciência da Informação do
IBICT, ou projetos específicos interinstitucionais, como
Projeto Genoma, poderão se integrar à BDB segundo esta
opção. Ela é ilustrada na figura 5. FIGURA 3 2001 29 Carlos Henrique Marcondes / Luís Fernando Sayão Carlos Henrique Marcondes / Luís Fernando Sayão FIGURA 4
Solução Técnica
[PROCESSOS DE HARVESTING] FIGURA 3 FIGURA 3 FIGURA 3 Provedores de dados: de uma forma ampla, seriam todas
as instituições brasileiras que possuem site Internet que
disponibiliza documentos eletrônicos em C&T. Eventualmente, este site abriga também um ambiente de
submissão/publicação de documentos em texto completo;
um autor registra seu documento no site através de um
conjunto de metadados e, opcionalmente, armazena aí seu
documento em formato eletrônico. O site do provedor de
dados provê facilidades de busca para acesso aos
documentos nele armazenados e, caso seja aderente ao
padrão OpenArchives Harvest Protocol, permitirá
também que os metadados dos documentos do seu acervo
sejam visíveis a um programa de harvest. Exemplos típicos
seriam o SCIELO, os diversos arquivos eletrônicos
existentes no mundo, como o CogPrints (http://
cogprints.soton.ac.uk/), ou o arquivo aberto do IMPA
(http://www.preprint.impa.br/indexEngl.html). interface única do portal, aos diferentes recursos
informacionais que comporão a BDB e conjunto de
metadados que descreverão e fornecerão uma visão
unificada dos diferentes conjuntos de documentos. Com relação aos mecanismos de submissão de consultas,
a BDB deverá incorporar as principais alternativas
tecnológicas discutidas anteriormente, buscas distribuídas
e busca em uma base centralizada de metadados obtida
mediante coleta automática (harvesting). Para isso, o portal
da BDB disporá de um programa cliente Z39.50, que lhe
permitirá acesso integrado por meio da distribuição de
consultas aos catálogos na Internet das principais
bibliotecas universitárias do país e estrangeiras servidas
pelo protocolo Z39.50. Qualquer outro recurso
informacional na Internet que seja servido por este
protocolo também poderá ser acessado do portal da BDB,
como, por exemplo, um arquivo de e-prints ou o Scielo, o
qual planeja implementar um servidor Z39.50 para sua
base. Esta opção é ilustrada na figura 3. Provedor de serviços: instituições que provêem serviços
de valor agregado sobre documentos eletrônicos
disponibilizados por um ou mais provedor de dados. Exemplos destes serviços seriam a montagem de bases de
dados qualificadas, o acesso unificado a documentos
armazenados em diferentes provedores de dados, revisão
e avaliação de documentos publicados em um ou mais
provedores de dados, linkagem de recursos informacionais. Exemplos típicos seriam o serviço Arc (http://
arc.cs.odu.edu/) que provê acesso unificado a diferentes
arquivos abertos e o objeto deste projeto, a própria BDB. Dado o papel integrador da proposta da BDB, seu modelo
de interoperabilidade se baseia em dois elementos:
mecanismos de submissão de consultas, a partir da Ci. Inf., Brasília, v. 30, n. 3, p. 24-33, set./dez. CONCLUSÕES – propor uma agenda incluindo os temas de pesquisa
citados anteriormente à comunidade de pesquisas
brasileira e uma linha de fomento correspondente,
envolvendo universidades, institutos de pesquisa e
instituições parceiras; – propor uma agenda incluindo os temas de pesquisa
citados anteriormente à comunidade de pesquisas
brasileira e uma linha de fomento correspondente,
envolvendo universidades, institutos de pesquisa e
instituições parceiras; As experiências internacionais em torno da questão das
publicações na rede e da interoperabilidade entre de
bibliotecas digitais são bastante recentes, contemporâneas
mesmo; as soluções para estes problemas são hoje o foco
das maiores atenções por parte dos pesquisadores de ciência
da informação e dos diversos sistemas de informação em
C&T. Apesar de toda esta ebulição, está patente que já
existe um conjunto de padrões e tecnologias maduros e
consolidados que são bases para vários sistemas de
informação importantes já em operação pelo mundo. As
questões de interoperabilidade endereçadas pelo projeto
da BDB são ainda bastante restritas, limitadas a um
enfoque tecnológico. A interoperabilidade entre recursos
informacionais heterogêneos na Internet tem várias outras
dimensões – semântica, política/humana, entre
comunidades, internacional, interlingüística (Powell,
1998) –, como alerta Miller. – propor um fórum nacional sobre o tema amplo de
bibliotecas digitais que sirva para discussão e troca de
experiências; – propor um fórum nacional sobre o tema amplo de
bibliotecas digitais que sirva para discussão e troca de
experiências; – propor um programa de formação de quadros” (IBICT,
2001, p.16). O projeto da BDB, embora tenha um compromisso
pragmático com a prestação de serviços à comunidade
acadêmica brasileira, coloca também questões relativas
ao planejamento de ICT no país. Desde fins da década da
de 80, com a Ação Programa de Informação em Ciência e
Tecnologia (Brasil, 1984) e com os PADCTs (Brasil, 1985),
não surgiram documentos abrangentes de planejamento
de ICT no Brasil. Os atuais Livros Verdes, o da Sociedade
da Informação (2000) e o de Ciência, Tecnologia e
Inovação (2001), não contemplam questões relativas à
ICT. O sucesso do projeto em uma perspectiva a médio e longo
prazo vai depender de o IBICT adotar uma nova postura
institucional com relação ao acompanhamento das
tendências e padrões tecnológicos envolvidos, à pesquisa,
ao desenvolvimento e à adaptação de tecnologias. Este
quadro é tão amplo que seria impossível ao IBICT
acompanhá-lo sozinho. O segundo elemento do esquema de interoperabilidade
da BDB é o conjunto de metadados. Ele deverá contemplar
e integrar, em uma descrição unificada, as diferentes
tipologias de documentos originários dos diferentes
recursos informacionais que comporão a BDB. A
referência emergente nesta área, avalizada pela
comunidade de informação, é Dublin Core Element Set. É resultado de intenso trabalho de discussão e
padronização em nível internacional, mantida por um
ativo grupo e fórum internacionais, a Dublin Core
Metadata Initiative, que já realizou diversos encontros; é
usado em diferentes sistemas, inclusive na OpenArchives
Initiative e se encontra em pleno desenvolvimento. elemento (Dublin Core, 2000) e pode ser codificado em
formatos como HTML e XML (Cox, 2000), (Beckett,
2000), (Beckett. 2001). Como é explicitado na própria
proposta Dublin Core, o conjunto de metadados deve ser
tão simples e intuitivo a ponto de permitir que o próprio
autor descreva seu trabalho. É exatamente assim que
funcionam os ambientes de submissão de trabalhos: ao
submeter seu trabalho, o autor preenche um formulário
com os metadados pertinentes. O conjunto de metadados Dublin Core é composto de 13
elementos descritivos (Dublin Core, 1999), suporta
qualificadores para especificar o significado de um Dada a sua característica integradora de diferentes recursos
informacionais heterogêneos, que armazenam diferentes
tipos de documentos, a BDB usará o conjunto Dublin Core, Ci. Inf., Brasília, v. 30, n. 3, p. 24-33, set./dez. 2001 30 Integração e interoperabilidade no acesso a recursos informacionais eletrônicos em C&T: a proposta da ... Integração e interoperabilidade no acesso a recursos informacionais eletrônicos em C&T: a proposta da heterogêneas distribuídas. É também de grande
importância a participação do IBICT nos principais fóruns
internacionais que discutem as questões relacionadas ao
tema. Sugere-se: expandido com qualificadores para suportar características
especiais de alguns tipos de documentos. Detalhes da
estrutura de metadados da BDB serão objeto de um
trabalho posterior. Ci. Inf., Brasília, v. 30, n. 3, p. 24-33, set./dez. 2001 REFERÊNCIAS BIBLIOGRÁFICAS E FONTES DE INFORMAÇÃO Um empreendimento amplo e com um caráter integrador
como a proposta da BDB deve ser gerido por um comitê
dirigente que inclua as parcerias estratégicas do IBICT
neste projeto e representantes da comunidade de C&T. Este Comitê Dirigente deve se reunir periodicamente para
analisar e aprovar o Relatório de Atividades da BDB e seu
Plano de Trabalho para o próximo período. Entre as
reuniões do comitê dirigente, a BDB será gerida por um
comitê executivo. ALLEN, J.; MEALLING, M. The architecture of the Common Indexing
Protocol (CIP). IETF Internet draft. Disponível em: <http://
search.ietf.org/internet-drafts/draft-ietf-cip-arch-02.txt>. Acesso em:
4 out. 2001. BECKETT, Dave; MILLER, Eric; BRICKLEY, Dan. An XML encoded
of simple Dublin core metadata. [S. l.] : Dublin Core Metadata
Initiative, 2001. Disponível em: <http://dublincore.org/documents/2001/
04/11/dcmes-xml>. Acesso em: 6 jun. 2001. BECKETT, Dave; MILLER, Eric; BRICKLEY, Dan.. Using Dublin core
in XML. [S. l.] : Dublin Core Metadata Initiative, 2000. Disponível
em:
<http://www.purl.org/dc/documents/wd/dcmes-xml-
20000714.htm>. Acesso em: 1 jun. 2001. Embora os objetivos da BDB sejam ambiciosos, mostram-
se também plenamente viáveis em termos tecnológicos;
a implantação da BDB pode se iniciar com um
investimento baixo, beneficiando-se da experiência e das
metodologias já desenvolvidas nos principais centros
internacionais. Estes objetivos também são necessários
para que a comunidade acadêmica brasileira possa dispor
de uma infra-estrutura informacional compatível com os
padrões internacionais. Isto permitirá ao país se inserir
plenamente nos fóruns científicos internacionais dentro
do paradigma atual da comunicação científica. E permitirá
principalmente que os sistemas de informação brasileiros
se integrem ao fluxo mundial de informações, dando maior
visibilidade à produção brasileira em C&T. O interesse
do pesquisador é conseguir a maior visibilidade possível
para sua produção acadêmica. Os serviços de informação
em C&T, entre eles as bibliotecas digitais como a BDB,
devem ajudar os autores dos documentos neles
armazenados a obter a máxima visibilidade da sua
produção, adotando mecanismos que maximizem a
integração e interoperabilidade amplas entre serviços de
informação. BERNERS-LEE, Tim; HENDLER, James; LASSILA, Ora. The semantic
web. Scientific American, New York, n. 5, May 2001. Disponível em:
<http://www.scian.com/2001/0501issue/0501berners-lee.html>. Aces-
so em: 24 maio 2001. BRASIL. Ministério da Ciência e Tecnologia. PADCT: documento base
[e] documentos sínteses dos subprogramas. Brasília : CNPq, 1985. 97 p. BRASIL. Presidência da Republica. Secretaria de Planejamento. III
PBDCT: III Plano Básico de Desenvolvimento Científico e Tecnológico:
informação em ciência e tecnologia. Brasília : CNPq, 1984, 69 p. (Ação
Programada em Ciência e Tecnologia). BRYAN, Martin. CONCLUSÕES Para fazer frente a este desafio, o
IBICT deve assumir o papel de articulador entre diferentes
parceiros nacionais e a comunidade acadêmica para um
trabalho conjunto em torno de uma agenda de pesquisas
que inclua questões de interesse do projeto da BDB. Para
isso, o projeto sugere: Hoje, no entanto, a comunicação científica é cada vez
mais fortemente dependente das tecnologias de
informação. O projeto da BDB menciona itens que
constituiriam a infra-estrutura necessária para viabilizar
um ambiente de informação integrado. Planejar e
implantar esta infra-estrutura seria claramente papel do
IBICT. Um ambiente como este garantiria aos usuários “a
informação na ponta dos dedos” e simultaneamente ampla
visibilidade à produção acadêmica brasileira. Componentes desta infra-estrutura seriam: ambientes
para submissão e armazenamento de documentos
eletrônicos, mecanismos de linkagem de documentos
eletrônicos entre si, de modo que um usuário pudesse ter
acesso imediatamente a referências e fontes citadas em
um documento eletrônico, endereços eletrônicos
persistentes, sem os problemas de links inválidos, base de
autoridades, linguagens de descrição, esquemas de
classificação temática e sistemas de metadados para
interoperabilidade entre sistemas e descoberta de
informações, armazenamento e preservação de
documentos eletrônicos por longo tempo. “Acompanhamento e articulação com os fóruns
internacionais que discutem questões como metadados,
interoperabilidade, publicações na rede, comunicação
científica via rede, preservação de documentos eletrônicos,
direitos autorais em documentos eletrônicos, linkagem de
recursos informacionais etc.” como World Wide Web
Consortiun, Dublin Core Metadata Initiative, Open
Archives Initiative, Digtal Library Federation. Com o
objetivo de acompanhar o desenvolvimento das
tecnologias associadas a bibliotecas digitais e o desenrolar
das controvérsias sobre estes assuntos e seus
desdobramentos, é necessária uma aproximação com as
organizações e/ou redes internacionais que operam e/ou
desenvolvem experiências com bibliotecas digitais 31 Ci. Inf., Brasília, v. 30, n. 3, p. 24-33, set./dez. 2001 Carlos Henrique Marcondes / Luís Fernando Sayão ntegração e interoperabilidade no acesso a recursos informacionais eletrônicos em C&T: a proposta da ... GINSPARG, P. Winners and losers in the global research village. In:
CONFERENCE ON ELECTRONIC PUBLISHING IN SCIENCE, 1996,
Paris. Proceedings... Disponível em: <http://xxx.lanl.gov/blurb/
pg96unesco.html>. Acesso em: 5 out. 2001. PAYETTE, Sandra; RIEGER, Oya Y. Z39.50: the user’s perspective. D-
Lib Magazine, Apr. 1997. Disponível em: <http://www.dlib.org/dlib/
april97/cornell/04payette.html>. Acesso em: 8 set. 2000. PINHEIRO, Lena Vânia Ribeiro; LOUREIRO, José Mauro Matheus. Traçados e limites da ciência da informação. Ciência da Informação,
Brasília, v. 24, n. 1, p. 42-53, jan./abr. 1995. GONÇALVES, Marcos Andre; FRANCE, Robert K.; FOX, Edward A. MARIAN: flexible interoperability for federated digital libraries. In:
ECDL-01: EUROPEAN CONFERENCE ON RESEARCH AND
ADVANCED TECHNOLOGY FOR DIGITAL LIBRARIES, 5., 2001,
Darmstadt. Proceedings... Disponível em: <http://www.dlib.vt.edu/
reports/ecdl2001_8.pdf>. Acesso em: 5 out. 2001. POWELL, James. Multilingual federated searching across
heterogeneous collections. D-Lib Magazine, Sept. 1998. Disponível em:
<http://www.dlib.org/dlib/september98/power//09power.html>. Acesso
em: 25 jul. 2001. GREENSTEIN, Daniel. The arts and humanities data service: three
years’ on. D-Lib Magazine, Dec. 1998. Disponível em: <http://
www.dlib.org/dlib/december98/greensteisn/greenstein.html>. Acesso
em: 1 jul. 2001. RIFKIN, Adam. A look at XML. WebDeveloper.com. Disponível em:
<http://webdeveloper.internet.com/xml/xml_a_look_at_xml.html>. Acesso em: 5 set. 2000. SHNEIDERMAN, Ben; BYRD, Don; CROFT, W. Bruce. Clarifying
search: a user-interface framework for text searches. D-Lib Magazine,
Jan. 1997. Disponível em: <http://www.dlib.org/dlib/january97/retrieval/
01sneiderman.html>. Acesso em: 5 out. 2001 THE IMESH toolkit: an architecture and toolkit for distributed subject
gateways. International Digital Libraries, n. 19, Jan. 1999. NSF 99-6. INSTITUTO BRASILEIRO DE INFORMAÇÃO EM CIÊNCIA E
TECNOLOGIA (Brasília, DF). Projeto técnico da biblioteca digital brasilei-
ra em C&T. Brasília, 2001. 40 p. SOCIEDADE da informação no Brasil : livro verde. Brasília : Ministé-
rio da Ciência e Tecnologia, 2000. (organizado por Tadao Takahashi.). LAWRENCE, Steve. Free online availability substantially increases a
paper’s impact. Disponível em: http://wwwnature.com/nature/debates/
e-access/Articles/lawrence.html. Acesso em: 10 jun. 2001. SOMPEL, Herbert van de; LAGOZE, Carl. The Santa Fe convention
of the open archives initiative. D-Lib Magazine, v. 6, n. 2, Feb. 2000. Disponível em : <http://www.dlib.org/dlib/february00/vandesompel-oai/
vandesompel-oai.html>. Acesso em: 5 out. 2001 LEVAN, Ralf. Dublin core and Z39.50. [S. l.] : Dublin Core Metadata
Initiative, 1998. Disponível em: <http://dublincore.org/documents/1998/
02/02/dc-z3950/>. Acesso em: 9 mar. 2001. SULEMAN, Hussein, et al. Networked digital library of theses and
dissertations: bringing the gap for global access: part 1: mission and
progress. D-Lib Magazine, v. 7, n. 9, Sept. 2001. Disponível em: <http:/
/www.dlib.org/dlib/september01/suleman/09suleman-pt1.html>. Aces-
so em: 19 set. 2001. LIU, Xiaoming et al. Arc: an OAI service provider for digital library
federation. Artigo recebido em 10/11/2001. REFERÊNCIAS BIBLIOGRÁFICAS E FONTES DE INFORMAÇÃO An introduction to Extensible Markup Language
(XML). [S. l.] : SGML Center, 1997. Disponível em: <http://
www.personal.u-net.com/~sgml/xmlintro.html>. Acesso em: 21 out. 2000. CIÊNCIA, tecnologia e inovação: desafio para a sociedade brasileira:
livro verde. Brasília : Ministério da Ciência e Tecnologia, Academia
Brasileira de Ciências, 2001. CLEVER PROJECT. Hypersearching the web. Scientific American, New
York, n. 6, 1999. Disponível em: <http://www.sciam.com/1999/0699issue/
0699raghavan.html>. Acesso em: 31 maio 1999. COX, Simon; MILLER, Eric; POWELL, Andy. Recording qualified
Dublin core metadata in HTML meta elements. [S. l.] : Dublin Core
Initiative, [2001?]. Disponível em: <http://dublincore.org/documents/
2000/08/15/dcq-html/>. Acesso em: 6 jul. 2001. DAVIS, James R. Creating a networked computer science technical
report library. D-Lib Magazine, Sept. 1995. Disponível em: <http://
www.dlib.org/dlib/september95/09davis.html>. Acesso em :16 jul. 2001. DIENST architecture summary description. Ithaca : Cornell University,
2000. Disponível em: <http://www.cs.cornell.edu/cdlrg/dienst/
architecture/architetucture.htm>. Acesso em: 5 out. 2001. DIENST protocol version 4.1. Draft. Ithaca : Cornell University, 1998. Disponível em: <http://www.cs.cornell.edu/NCSTRL/protocol.htm>. Acesso em: 5 out. 2001. DIENST software summary description. Ithaca : Cornell University,
2000. Disponível em: <http://www.cs.cornell.edu/cdlrg/dienst/software/
DienstSoftware.htm>. Acesso em: 5 out. 2001. DUBLIN core metadata elements set, version 1.1: reference description. [S. l.] : Dublin Core Initiative, 1999. Disponível em: <http://
dublincore.org/documents/1999/07/02/dces/>. Acesso em: 6 jun. 2001. DUBLIN core qualifiers. [S. l.] : Dublin Core Initiative, 2000. Dispo-
nível em: <http://purl.org/dc/documents/rec/dcmes-qualifiers-
20000711.htm>. Acesso em: 5 out. 2001. EXTENSIBLE markup language (XML). [S. l.] : World Wide Web
Consortium, 2000. Disponível em: <http://www.w3.org/XML/>. Aces-
so em: 30 jan. 2001. 32 Ci. Inf., Brasília, v. 30, n. 3, p. 24-33, set./dez. 2001 Ci. Inf., Brasília, v. 30, n. 3, p. 24-33, set./dez. 2001 ntegração e interoperabilidade no acesso a recursos informacionais eletrônicos em C&T: a proposta da ... D-Lib Magazine, v. 7, n. 4, Apr. 2001. Disponível em: <http:/
/www.dlib.org/dlib/april01/liu/04liu.html>. Acesso em: 25 jul. 2001. LYNCH, Clifford. The Z39.50 information retrieval standard. D-Lib
Magazine, v. 7, n. 4, Apr. 2001. Disponível em: <http://www.dlib.org/
dlib/april97/liu/04lynch.html>. Acesso em: 16 mar. 2001. SULEMAN, Hussein, et al. Networked digital library of theses and
dissertations: bringing the gap for global access: part 2: services and
research. D-Lib Magazine, v. 7, n. 9, Sept. 2001. Disponível em: <http:/
/www.dlib.org/dlib/september01/suleman/09suleman-pt2.html>. Aces-
so em: 19 set. 2001. MEDEIROS, Norm. XML and the resource description framework:
the great web hope. Online, v. 24, n. 5, Sept. 2000. Disponível em:
<http://www.onlineinc.com/onlinemag/OL2000/medeiros9.html>. Acesso em: 9 jan. 2001. TROLL, Denise; MOEN, Bill. Report to the DLF on the Z39.50
Implementers’ Group. DLF : 2001. Disponível em: <http://
www.diglib.org/architectures/zig0012.htm>. Acesso em: 22 ago. 2001. MILLER, Paul. UK interoperability focus: Bath, UK, UKOLN. Dispo-
nível em: <http://www.ukoln.ac.uk/interop-focus/about/>. Acesso em:
3 out. 2001. XML schema part 0: primer. W3C Working Draft, 7 April 2000. Dispo-
nível em: <http://www.w3.org/TR/2000/WD-xmlschema-0-20000407/>. Acesso em: 5 set. 2000. NETWORK WORKING GROUP. Architecture of the whois++ service. RFC 1835.. Disponível em: <http://src.doc.ic.ac.uk/computing/internet/
rfc/rfc1835.txt>. Acesso em: 4 out. 2001. Z39.50 profiles. Disponível em: <http://locweb.loc.gov/z3950/agency/
profiles/profiles.html>. Acesso em: 30 mar. 2001. PAEPKE, Andreas et al. Search middleware and the simple digital
library interoperability protocol . D-Lib Magazine, v. 6, n. 3, Mar. 2000. Disponível em: <http://www.dlib.org/dlib/march00/paepcke/
03paepcke.html>. Acesso em: 26 jul. 2001. Ci. Inf., Brasília, v. 30, n. 3, p. 24-33, set./dez. 2001 33
|
https://openalex.org/W2140282099
|
https://repositorio.unican.es/xmlui/bitstream/10902/26542/2/AComparisonOf_nv_articulo_15119.pdf
|
English
| null |
A comparison of algorithms for the construction of SZ cluster catalogues
|
Astronomy & astrophysics
| 2,012
|
cc-by
| 13,589
|
Astronomy
&
Astrophysics Astronomy
&
Astrophysics Astronomy
&
Astrophysics A&A 548, A51 (2012)
DOI: 10.1051/0004-6361/201015689
c⃝ESO 2012 A&A 548, A51 (2012)
DOI: 10.1051/0004-6361/201015689
c⃝ESO 2012 A comparison of algorithms for the construction
of SZ cluster catalogues Thomson Avenue, Cambridge, CB3 0HE,
9 9 Max-Planck-Institut für extraterrestrische Physik Giessenbachstr, 85748 Garching, Germany 10 Instituto de Física de Cantabria (CSIC-UC), 39005 Santander, Spain
11 10 Instituto de Física de Cantabria (CSIC-UC), 39005 Santander, Spain
11 Kavli Institute for Cosmology Cambridge and Institute of Astronomy, Madingley Road, Cambridge, CB3 OHA, UK
2 11 Kavli Institute for Cosmology Cambridge and Institute of Astronomy, Madingley Road, Cambridge, CB3 OHA, UK
12
l
lf
i
d
i
l
d
d
i
ill 12 ALMA JAO, Av. El Golf 40 – Piso 18, Las Condes, Santiago; ESO, Alonso de Córdova 3107, Vitacura, Casilla
Santiago, Chile 13 Università di Roma “Tor Vergata”, via della ricerca scientifica, 1, 00133 Roma, Italy
14 Astronomisches Rechen Institut, Monchhofstrasse 12 14, 69120 Heidelberg, Germany
15 Dipartimento di Astronomia, Università di Bologna, via Ranzani 1, 40127 Bologna, Italy Received 3 September 2010 / Accepted 3 October 2012 Received 3 September 2010 / Accepted 3 October 2012 ABSTRACT We evaluate the construction methodology of an all-sky catalogue of galaxy clusters detected through the Sunyaev-Zel’dovich (SZ)
effect. We perform an extensive comparison of twelve algorithms applied to the same detailed simulations of the millimeter and
submillimeter sky based on a Planck-like case. We present the results of this “SZ Challenge” in terms of catalogue completeness,
purity, astrometric and photometric reconstruction. Our results provide a comparison of a representative sample of SZ detection
algorithms and highlight important issues in their application. In our study case, we show that the exact expected number of clusters
remains uncertain (about a thousand cluster candidates at |b| > 20 deg with 90% purity) and that it depends on the SZ model and on the
detailed sky simulations, and on algorithmic implementation of the detection methods. We also estimate the astrometric precision of
the cluster candidates which is found of the order of ∼2 arcmin on average, and the photometric uncertainty of about 30%, depending
on flux. Key words. cosmology: observations – galaxies: clusters: general – galaxies: clusters: intracluster medium –
cosmic background radiation – methods: data analysis A comparison of algorithms for the construction
of SZ cluster catalogues J.-B. Melin1, N. Aghanim2, M. Bartelmann3, J. G. Bartlett4,5,6, M. Betoule4, J. Bobin7, P. Carva J.-B. Melin1, N. Aghanim2, M. Bartelmann3, J. G. Bartlett4,5,6, M. Betoule4, J. Bobin7, P. Carvalho8, G. Chon9,
J. Delabrouille4, J. M. Diego10, D. L. Harrison11, D. Herranz10, M. Hobson8, R. Kneissl12, A. N. Lasenby8,11,
M. Le Jeune4, M. Lopez-Caniego10, P. Mazzotta13, G. M. Rocha6, B. M. Schaefer14, J.-L. Starck7,
J. C. Waizmann3,15, and D. Yvon1 J. Delabrouille4, J. M. Diego10, D. L. Harrison11, D. Herranz10, M. Hobson8, R. Kneissl12, A. N. Lasenby8,11,
M. Le Jeune4, M. Lopez-Caniego10, P. Mazzotta13, G. M. Rocha6, B. M. Schaefer14, J.-L. Starck7,
J. C. Waizmann3,15, and D. Yvon1 1 DSM/Irfu/SPP, CEA/Saclay, 91191 Gif-sur-Yvette Cedex, France 1 DSM/Irfu/SPP, CEA/Saclay, 91191 Gif-sur-Yvette Cedex, France e-mail: jean-baptiste.melin@cea.fr e mail: jean baptiste.melin@cea.fr
2 Institut d’Astrophysique Spatiale, CNRS et Université de Paris XI, Bâtiment 120-121, Centre Universitaire d’Orsay,
91400 Orsay Cedex, France j
p
2 Institut d’Astrophysique Spatiale, CNRS et Université de Paris XI, Bâtiment 120-121, Centre Universitaire d’Orsay,
91400 Orsay Cedex, France y
3 Institut fur Theoretische Astrophysik, Zentrum fur Astronomie der Universitat Heidelberg, Albert-Ueberle-Str. 2
69120 Heidelberg, Germany y
3 Institut fur Theoretische Astrophysik, Zentrum fur Astronomie der Universitat Heidelberg, Albert-Ueberle-Str. 2,
69120 Heidelberg, Germany
4 4 APC, 10 rue Alice Domon et Léonie Duquet, 75205 Paris Cedex 13, France
5 4 APC, 10 rue Alice Domon et Léonie Duquet, 75205 Paris Cedex 13, France
5 5 Université Paris Diderot – Paris 7, 75205 Paris Cedex 13, France
6 Université Paris Diderot – Paris 7, 75205 Paris Cedex 13, F 5 Université Paris Diderot – Paris 7, 75205 Paris Cedex 13, France
6
l i
b
lif
i
i
f
h
l
k
i
d 5 Université Paris Diderot – Paris 7, 75205 Paris Cedex 13, France
6 Jet Prop lsion Laborator
California Instit te of Technolog
4800 Oak Gro e Dri e Pasadena CA 91109 US 6 Jet Propulsion Laboratory, California Institute of Technology, 4800 Oak Grove Drive, Pasadena, CA 91109, USA
7 6 Jet Propulsion Laboratory, California Institute of Technology, 4800 Oak Grove Drive, Pasadena, CA 91
7 6 Jet Propulsion Laboratory, California Institute of Technology, 4800 O 7 DSM/Irfu/SAp, CEA/Saclay, 91191 Gif-sur-Yvette Cedex, France 7 DSM/Irfu/SAp, CEA/Saclay, 91191 Gif-sur-Yvette Cedex, France 8 Astrophysics Group, Cavendish Laboratory, J.J. 2.1. The Planck Sky Model One of the important differences between the present work
and previous studies of Planck SZ capabilities is the detailed
and rather sophisticated simulation of millimeter and submil-
limeter sky emission used here. Our sky simulations are based
on an early development version of the PSM (Delabrouille
et al. 2012), a flexible software package developed within the
Planck Collaboration for making predictions, simulations and
constrained realisations of the microwave sky. The simulations
used for this challenge are not polarised (only temperature maps
are useful for detecting clusters using the thermal SZ effect). Spinning dust emission uses as a template the dust extinction
map E(B −V), and uses an emission law uniform on the sky,
based on the model of Draine & Lazarian (1998), assuming a
warm neutral medium (WNM). We emphasise that the emission laws of both synchrotron
and dust vary across the sky. The spectral index of free-free and
the emission law of spinning dust, however, are taken as uniform
on the sky. The CMB sky is based on the best fit angular power spectrum
model of WMAP. The CMB realisation is not constrained by
actual observed CMB multipoles, in contrast to the simulations
used by Leach et al. (2008). y
Point sources are modeled with two main categories: radio
and infrared. In the present simulation, none of two is corre-
lated with the SZ signal. Simulated radio sources are based on
the NVSS or SUMSS and GB6 or PMN catalogues. Measured
fluxes at 1 and/or 4.85 GHz are extrapolated to 20 GHz using
their measured SED when observed at two frequencies. Sources
for which a flux measurement is available at a single frequency
have been randomly assigned to either the steep- or to the flat-
spectrum class in the proportions observationally determined by
Fomalont et al. (1991) for various flux intervals, and assigned
a spectral index randomly drawn from the corresponding distri-
bution. Source counts at 5 and 20 GHz obtained in this way are
compared, for consistency, with observed counts, with the model
by Toffolatti et al. (1998), and with an updated version of the
model by de Zotti et al. (2005), allowing for a high-redshift de-
cline of the space density of both flat-spectrum quasars (FSQs)
and steep-spectrum sources (not only for FSQs as in the origi-
nal model). 1. Introduction to higher redshifts with vastly superior statistics and control of
systematics. Galaxy cluster catalogues have played a long-standing, vital role
in cosmology, providing important information on topics rang-
ing from cosmological parameters to galaxy formation (Rosati
et al. 2002; Voit 2005). In particular, recent X-ray cluster cat-
alogues have proved valuable in establishing the standard cos-
mological model (e.g., Schuecker et al. 2003; Vikhlinin et al. 2009). The science potential of large cluster surveys is strong:
They are, for instance, considered one of the central observa-
tional tools for illuminating the nature of dark energy (e.g., the
Dark Energy Task Force Report Albrecht et al. 2006, 2009). A
suite of large cluster surveys planned over the coming years in
the optical/IR, X-ray and millimeter bands will greatly extend
the reach of cluster science by probing much larger volumes The Planck SZ cluster catalogue will be one of the impor-
tant players in this context. Surveying the entire sky in 9 mil-
limeter/submillimeter bands with ∼5–10 arcmin resolution over
the channels most sensitive to the cosmic microwave back-
ground (CMB) anisotropies, the Planck satellite will find large
numbers of clusters through the Sunyaev-Zel’dovich (SZ) effect
(Sunyaev & Zeldovich 1970, 1972; Birkinshaw 1999; Carlstrom
et al. 2002). The advantages of this, much anticipated, tech-
nique include efficient detection of distant clusters and selection
based on an observable expected to correlate tightly with clus-
ter mass (Bartlett 2002; da Silva et al. 2004; Motl et al. 2005;
Nagai 2006; Bonaldi et al. 2007; Shaw et al. 2008). An official
mission deliverable, the Planck SZ catalogue will be the first Article published by EDP Sciences A51, page 1 of 13 A&A 548, A51 (2012) all-sky cluster catalogue since the workhorse catalogues from
the ROSAT All-Sky Survey (RASS, Truemper 1992), in other
words, Planck will be the first all-sky cluster survey since the
early 1990s! were added in both templates to match the Planck resolution. The free-free emission law is constant over the sky, as it de-
pends only on the electronic temperature, taken as a constant
here (see however Wakker et al. 2008, for a description of high-
velocity clouds not detected by the WHAM survey and hence
not included in our simulations). Within the Planck Consortium, a considerable effort has been
conducted for the scientific evaluation of the cluster catalogue
construction methodology. 1. Introduction As part of this evaluation effort, we
completed an extensive comparison of twelve algorithms applied
to detailed simulations of Planck data based on the Planck Sky
Model (PSM). This study was dubbed “The SZ Challenge” and
was carried out in two steps using different SZ cluster models
and cosmologies; these are referred to as Versions 1 and 2 and
more fully explained below. We report the findings of these ini-
tial studies in terms of catalogue completeness and purity, as
well as astrometric and photometric accuracy and precision. Synchrotron emission is based on an extrapolation of the
408 MHz map of Haslam et al. (1982) from which an estimate
of the free-free emission was removed. In any direction on the
sky, the spectral emission law of the synchrotron is assumed to
follow a power law, T sync
b
∝νβ. We use a pixel-dependent spec-
tral index β derived from the ratio of the 408 MHz map and the
estimate of the synchrotron emission at 23 GHz in the wmap
data obtained by Miville-Deschênes et al. (2008). The thermal emission from interstellar dust is estimated us-
ing model 7 of Finkbeiner et al. (1999). This model, fitted to the
FIRAS data (7◦resolution), assumes that each line of sight can
be modeled by the sum of the emission from two dust popula-
tions, one cold and one hot. Each grain population is in thermal
equilibrium with the radiation field and thus has a grey-body
spectrum, so that the total dust emission is modelled as The article is organized as follows: In the next section we
detail our sky simulations, including a brief description of their
basis, the PSM (Delabrouille et al. 2012). The following section
then introduces the different catalogue construction methodolo-
gies employed, before moving on to a presentation of each of
the twelve algorithms in the study. We present the results of the
challenge in Sect. 4, followed by a comparative study of algo-
rithmic performance. We conclude with a discussion of both the
limitations of this study and future directions. Iν ∝
2
i=1
fiνβiBν(Ti),
(1) Iν ∝
2
i=1
fiνβiBν(Ti), (1) where Bν(Ti) is the Planck function at temperature Ti. In model 7
the emissivity indices are β1 = 1.5, β2 = 2.6, and f1 = 0.0309
and f2 = 0.9691. 1. Introduction Once these values are fixed, the dust temper-
ature of the two components is determined using only the ratio
of the observations at 100 μm and 240 μm. For this purpose,
we use the 100/240 μm map ratio published by Finkbeiner et al. (1999). Knowing the temperature and β of each dust component
at a given position on the sky, we use the 100 μm brightness at
that position to scale the emission at any frequency using Eq. (1). 2.2.2. SZ challenge v2 The second version of the SZ Challenge was produced using
a WMAP5 only cosmology (Dunkley et al. 2009) (h = 0.719,
Ωm = 0.256, ΩΛ = 0.744, σ8 = 0.798). We used the Jenkins
mass function (Jenkins et al. 2001). The SZ emission is mod-
eled using the universal pressure profile derived from the X-ray
REXCESS cluster sample (Arnaud et al. 2010) which predicts
profile and normalisation of SZ clusters given their mass and
redshift. The profile is well fitted by a generalized NFW profile
that is much steeper than the β-profile in the outskirts. Moreover,
for a given mass, the normalisation of the SZ flux is ∼15% lower
than the normalisation of SZ Challenge v1. This profile was used
as the baseline profile in the SZ early results from Planck (Planck
Collaboration 2011b,f,e,c,g). The SZ component is described in detail in the following
section. Component maps are produced at all Planck central frequen-
cies. They are then co-added and smoothed with Gaussian beams
as indicated in Table 1, extracted from the Planck blue book. We
thus obtain a total of nine monochromatic sky maps. Finally, inhomogeneous noise is simulated according to the
pixel hit count corresponding to a nominal 14-month mission1
using the Level-S simulation tool (Reinecke et al. 2006). The
rms noise level in the simulated maps is given in Table 1 from
the Planck blue book. It is worth noting that the in-flight perfor-
mances of Planck-HFI as reported by the Planck Collaboration
(Planck Collaboration 2011a) are better than the requirements. Neither of the two sets of simulations (v1 and v2) contains
point sources within clusters. The effect of contamination by ra-
dio or infrared point sources in clusters was therefore not stud-
ied here3. We neither include relativistic electronic populations
within clusters. As for point sources, this effect was not studied
here4. 3. Methods and algorithms The SZ Challenge was run as a blind test by providing the simu-
lated sky maps. Participating teams, ten, were then asked to run
their algorithms, twelve in total on the simulated data and supply
a cluster catalogue with 1. (α, δ): cluster sky coordinates; 2. Yrec: recovered total SZ flux, in terms of the integrated
Compton-Y parameter; s of instrumental values taken from Planck blue book for a 14 month nominal mission. Channel
30 GHz
44 GHz
70 GHz
100 GHz
143 GHz
217 GHz
353 GHz
545 GHz
857 GHz
FWHM [arcmin]
33
24
14
10
7.1
5
5
5
5
σpixel [mKCMB]
0.131
0.130
0.126
0.057
0.029
0.046
0.137
1.241
56.639
Notes. σpixel refers to the standard deviation of the 1.7′ (Healpix nside = 2048) pixel noise maps at the considered frequency. the spherical collapse model. The temperature of each cluster is
derived using a mass-temperature given in Colafrancesco et al. (1997) with T15 = 7.75 keV, consistent with the simulations of
Eke et al. (1998). For more details on this model, we refer the
reader to Sect. 5 of de Zotti et al. (2005). Infrared sources are based on the iras catalogue, and modeled
as dusty galaxies (Serjeant & Harrison 2005). IRAS coverage
gaps were filled by randomly adding sources with a flux distri-
bution consistent with the mean counts. Fainter sources were as-
sumed to be mostly sub-millimeter bright galaxies, such as those
detected by SCUBA surveys. These were modelled following
Granato et al. (2004) and assumed to be strongly clustered, with
a comoving clustering radius r0 ≃8 h−1 Mpc. Since such sources
have a very high areal density, they are not simulated individu-
ally but make up the sub-mm background. 2.2. The SZ component We simulate the SZ component using a semi-analytic approach
based on an analytic mass function dN(M, z)/dMdz. After select-
ing cosmological parameters (h, Ωm, ΩΛ, σ8), the cluster distri-
bution in the mass-redshift plane (M, z) is drawn from a Poisson
law whose mean is given by the mass function. Clusters are span-
ning the mass range 5 × 1013 M⊙< Mvir < 5 × 1015 M⊙and the
redshift range 0 < z < 4. Cluster Galactic coordinates (l, b) are
then uniformly drawn on the sphere. We compute the SZ signal
for each dark matter halo following two different models, pro-
ducing two simulations (v1 and v2) with different sets of cosmo-
logical parameters and mass functions and SZ signals2. 1 Note that since launch in May 2009, the observed Helium consump-
tion for the Planck-HFI dilution cooler indicates that the instrument can
operate for about 30 months. A mission extension has been approved
by ESA accordingly. 2 We assume spherical symmetry for the individual SZ clusters and do
not take into account any scatter in the distribution of pressure profiles. 3 The effect of radio sources (ν < 217 GHz) is to reduce the ob-
served SZ signal at a given frequency while the effect of infrared sources
(ν > 217 GHz) is to increase it. However, the extraction algorithms be-
ing multifrequency, their sensitivity to point sources is expected to be
weaker than for single frequency extractions because of the different
spectral dependence of point sources and SZ clusters.
4 The effect has been very recently studied within the Planck
Collaboration: assuming a non relativistic spectrum for extracting clus-
ters biases the flux low by about 10% in direction of massive (Mvir >
1015 M⊙) clusters (Planck Collaboration, in prep.). 4 The effect has been very recently studied within the Planck
Collaboration: assuming a non relativistic spectrum for extracting clus-
ters biases the flux low by about 10% in direction of massive (Mvir >
1015 M⊙) clusters (Planck Collaboration, in prep.). 1 Note that since launch in May 2009, the observed Helium consump-
tion for the Planck-HFI dilution cooler indicates that the instrument can
operate for about 30 months. A mission extension has been approved
by ESA accordingly.
2 We assume spherical symmetry for the individual SZ clusters and do
not take into account any scatter in the distribution of pressure profiles. 2.1. The Planck Sky Model Further extrapolation at Planck frequencies has been
made allowing a change in SED above 20 GHz, assuming again
a distribution in flat and steep populations. For each of these
two populations, the spectral index is randomly drawn within a
set of values compatible with the typical average and dispersion. Diffuse Galactic emission is described by a four compo-
nent model of the interstellar medium comprising free-free,
synchrotron, thermal dust and spinning dust and is based on
Miville-Deschênes et al. (2008, see Miville-Deschênes 2009, for
a review). The predictions rely on a number of sky templates
with different angular resolutions. In order to simulate the sky
at Planck resolution we have added small-scale fluctuations to
some of the templates to increase the fluctuation level as a func-
tion of the local brightness and therefore reproduce the non-
Gaussian and non-uniform properties of the interstellar emis-
sion. The procedure used to add small scales is presented in
Miville-Deschênes et al. (2007). Free-free emission is based on the model of Dickinson et al. (2003) assuming an electronic temperature of 7000 K. The spa-
tial structure of the emission is modeled using an Hα template
corrected for dust extinction. The Hα map is a combination of
the Southern H-Alpha Sky Survey Atlas (SHASSA, Gaustad
et al. 2001) and the Wisconsin H-Alpha Mapper (WHAM,
Haffner et al. 2003), smoothed to obtain a uniform angular reso-
lution of 1◦. Dust extinction is inferred using the E(B−V) all-sky
map of Schlegel et al. (1998). As mentioned earlier, small scales A51, page 2 of 13 J.-B. Melin et al.: A comparison of algorithms for the construction of SZ cluster catalogues 2.2.1. SZ challenge v1 3. ΔYrec: estimated flux error, i.e., the method’s internal esti-
mate of flux error; For the first version of the SZ Challenge, we used h = 0.7,
Ωm = 0.3, ΩΛ = 0.7, σ8 = 0.85 and the Sheth-Tormen mass
function Sheth & Tormen (1999). We assume that the clusters
are isothermal and that the electron density profile is given by
the β-model, with β = 2/3, and core radius scaling as M1/3. We
truncate the model at the virial radius, rvir, and choose the core
radius rc = rvir/10. The virial radius here is defined according to 4. θrec: recovered cluster angular size, in terms of equivalent
virial radius; 5. Δθrec: estimated size error (internal error). 3 The effect of radio sources (ν < 217 GHz) is to reduce the ob-
served SZ signal at a given frequency while the effect of infrared sources
(ν > 217 GHz) is to increase it. However, the extraction algorithms be-
ing multifrequency, their sensitivity to point sources is expected to be
weaker than for single frequency extractions because of the different
spectral dependence of point sources and SZ clusters. A51, page 3 of 13 A&A 548, A51 (2012) The different methods were divided into two classes: direct
methods that produce a cluster catalogue applying filters directly
to a set of frequency maps, and indirect methods that first extract
a thermal SZ map and then apply source finding algorithms. The different methods were divided into two classes: direct
methods that produce a cluster catalogue applying filters directly
to a set of frequency maps, and indirect methods that first extract
a thermal SZ map and then apply source finding algorithms. using a linear combination of maps (which requires an estimate
of the statistics of the contamination) and uses spatial filtering
to suppress both foregrounds and noise (making use of prior
knowledge of the cluster profile). In all cases discussed here, the
adopted template is identical to the simulated cluster profiles, ex-
cept for MMF2 on SZ Challenge v2. The MMF has been studied
extensively by Herranz et al. (2002) and Melin et al. (2006). q
y
p ,
a thermal SZ map and then apply source finding algorithms. In this classification, the 12 algorithms studied were: – Direct methods: • MMF1: matched multi-filters (MMF) as implemented by
Harrison. 5 Needlet coefficients are the equivalent of Fourier coefficients in the
adopted spherical wavelet domain. 2.2.1. SZ challenge v1 Three of the MMF methods tested here work with projected
flat patches of the sky, and one method works directly on the
pixelised sphere. • MMF2: MMF as described in Herranz et al. (2002). • MMF3: MMF as described in Melin et al. (2006). In the first case, the full-sky frequency maps are projected
onto an atlas of overlapping square flat regions. The filtering is
then implemented on sets of small patches comprising one patch
for each frequency channel. For each such region, the nine fre-
quency maps are processed with the MMF. A simple threshold-
ing detection algorithm is used to find the clusters and produce
local catalogues. The MMF is applied with varying cluster sizes
to find the best detection for each cluster. This provides an esti-
mate of the angular size in addition to the central Compton pa-
rameter. Each algorithm explored its own, but similar, range of
angular scales; MMF3, for example, runs from θv = 2 to 150 ar-
cmins. The catalogues extracted from individual patches are then
merged into a full-sky catalogue that contains the position of the
clusters, their estimated central Compton parameter, the virial
radius and an estimation of the error in the two later quantities. The integrated Compton parameter is derived from the value of
the central Compton parameter and the radius of the cluster. • MMF4: MMF as described in Schäfer et al. (2006). • PwS: Bayesian method PowellSnakes as described
in Carvalho et al. (2009). Indirect methods: • BNP: Bayesian Non-parametric method as described
in Diego et al. (2002), followed by SExtractor (Bertin
& Arnouts 1996) to detect clusters and perform
photometry. • ILC1: All-sky internal linear combination (ILC) on
needlet coefficients5 (similar to the method used for
CMB extraction in Delabrouille et al. 2009) to get an
SZ map, followed by matched filters on patches to ex-
tract clusters and perform photometry. • ILC2: Same SZ map Delabrouille et al. (2009), but
followed by SExtractor on patches instead of a matched
filter to extract the clusters. Photometry or flux measure-
ment is however done using matched filters at the posi-
tion of the detected clusters. In the following, we give relevant details specific to each im-
plementation of the MMF. • ILC3 developed by Chon and Kneissl: ILC in real space
and filtering in harmonic space to obtain an SZ map, fol-
lowed by fitting a cluster model. 3.1.1. MMF1 y
g
• ILC4 developed by Melin: ILC on patches in Fourier
space to obtain SZ maps, followed by SExtractor to de-
tect clusters and matched multifilters to perform photom-
etry. The performance of the MMF1 algorithm is sensitive to the ac-
curacy of the evaluation of the power spectra and cross-power
spectra of the non-SZ component of the input maps. The de-
tection is performed in two passes, the first detecting the high-
est signal-to-noise SZ clusters, and the second detecting fainter
clusters after the removal of the contribution of the brightest ones
from the power spectra estimated on the maps. y
• ILC5 developed by Yvon: ILC on patches in wavelet
space to obtain SZ maps, followed by SExtractor to de-
tect clusters and perform photometry. p
p
y
• GMCA:
Generalized
Morphological
Component
Analysis as described in Bobin et al. (2008), followed
by wavelet filtering and SExtractor to extract clusters
and matched multifilters to perform photometry. The merging of the catalogues from distinct patches is im-
plemented with the option of discarding detections found in the
smallest radius bin. These detections essentially correspond to
spatial profiles indistinguishable from that of a point source. This option permits better control of the contamination by point
sources, as a disproportionate fraction of the spurious detections
occur in this bin ; despite their different spectral signature, point
sources can occasionally pass through the filter. Using this op-
tion of MMF1 reduces the contamination at a given threshold
(see Sect. 4.1 for definitions of this and other diagnostics of cata-
logue content) depending on the actual profile of the SZ clusters. All of the algorithms make use of the known frequency spec-
trum of the SZ signal; attempts to detect clusters without this
prior knowledge perform significantly worse. A summary of the
characteristics of the codes as well as their treatment of the point
sources, foreground removal and masking is given in Table 2. Further details about the algorithms are given in the following
subsections. 3.1.2. MMF2 The multifrequency matched filter (MMF) enhances the contrast
(signal-to-noise) of objects of known shape and known spectral
emission law over a set of observations containing correlated
contamination signals. It offers a practical way of extracting a
SZ clusters using multifrequency maps. The method makes use
of the universal thermal SZ effect frequency dependance (as-
suming electrons in clusters are non-relativistic), and adopts a
spatial (angular profile) template. The filter rejects foregrounds The MMF2 algorithm follows closely the method described in
Herranz et al. (2002). The method is simple and quite robust,
although the performance depends on the model assumed for
the radial profile of the clusters. For this work, a truncated
multiquadric profile similar to a β-model has been used for
SZ Challenge v1 and v2. The profile is not a good match for the
simulated profile in SZ Challenge v2. This does not affect sig-
nificantly the completeness and purity of the method (as shown
in Sect. 4) but the extracted flux is biased with respect to the A51, page 4 of 13 J.-B. Melin et al.: A comparison of algorithms for the construction of SZ cluster catalogues Table 2. Summary of the algorithms compared in the SZ Challenge. Method
Shape matching
CPU
Patches
Number of
PS subtraction
FG subtraction
Main characteristics
time (h)
(size deg.)
patches
method
method
MMF1
Yes
50–60
14.6◦× 14.6◦
640
–
–
Best yield among MMFs
Good photometry
MMF2
Yes
31
14.6◦× 14.6◦
371
–
–
Good yield
Good photometry
MMF3
Yes
5
10◦× 10◦
504
mask 10σ PS
–
Good yield
Good photometry
MMF4
Yes
full sky
–
–
–
Poor yield (see Sect. 3.1.4)
No photometry
PwS
Yes
5.73
14.6◦× 14.6◦
2064
–
–
Good yield
Good photometry
BNP
No
15
10◦× 10◦
512
MHW
Subtract 857
Median yield
Poor photometry
ILC1
Yes
2–3
see caption
(504)
–
–
Good yield
Good photometry
ILC2
No
2–3
see caption
(504)
–
–
Best yield among ILCs
Good photometry
ILC3
Yes
24
full sky
–
–
Template fitting
Poor yield
Good photometry
ILC4
Yesa
6
10◦× 10◦
504
mask 10σ PS
–
Good yield
Good photometry
ILC5
No
0.2
11◦× 11◦
461
SExt. –
Poor yield (see Sect. 3.3.5)
Poor photometry
GMCA
No
4
10◦× 10◦
504
–
–
Median yield
Good photometry
Notes. The first column shows the name of the method. 3.2.1. PowellSnakes PowellSnakes (PwS; Carvalho et al. 2009) is a fast multi-
frequency Bayesian detection algorithm. It analyses flat sky
patches using the ratio ρ ≡Pr(H1|d)
Pr(H0|d),
(2) ρ ≡Pr(H1|d)
Pr(H0|d), (2) 3.1.2. MMF2 Note, also, that the study is made only
on clusters at |b| > 20 degrees Galactic latitude for all methods. The eighth column summarizes the main characteritics of each algorithm in terms
of yield at 90% purity and photometric accuracy. Notes. The first column shows the name of the method. The second column indicates when the code is using a prior on the SZ cluster shape. The uperscript a indicate that the detection did not use a shape prior but that the computation of the SZ flux did. The third column gives the
performance in terms of CPU hours needed to complete the analysis. The fourth and fifth column show whether the analysis was made using
all-sky maps or projected patches, their area in square degrees and the number of patches. Methods ILC1 and ILC2 work with full sky maps for
producing an SZ map (by ILC on needlet coefficients) and then work with 504 small patches for cluster detection by matched filtering (ILC1) or
using SExtractor (ILC2). The sixth and seventh columns provide information about any specific method used for subtracting point sources (PS)
and Galactic foregrounds (FG). Note that both the MMF and the ILC methods have a built-in way for subtracting both point sources and diffuse
foregrounds, by treating them as additional noise (of astrophysical origin) correlated across the channels. Note, also, that the study is made only
on clusters at |b| > 20 degrees Galactic latitude for all methods. The eighth column summarizes the main characteritics of each algorithm in terms
of yield at 90% purity and photometric accuracy. input. The family of profiles used by the algorithm can however
be adjusted. input. The family of profiles used by the algorithm can however
be adjusted. like many of the other algorithms, although it is in principle pos-
sible to include that extension. The large noise contribution due
to the Galaxy is the principal reason why the performance of the
filter suffers in comparison to the approach of discarding a large
fraction of the sky. 3.1.2. MMF2 The second column indicates when the code is using a prior on the SZ cluster shape. The uperscript a indicate that the detection did not use a shape prior but that the computation of the SZ flux did. The third column gives the
performance in terms of CPU hours needed to complete the analysis. The fourth and fifth column show whether the analysis was made using
all-sky maps or projected patches, their area in square degrees and the number of patches. Methods ILC1 and ILC2 work with full sky maps for
producing an SZ map (by ILC on needlet coefficients) and then work with 504 small patches for cluster detection by matched filtering (ILC1) or
using SExtractor (ILC2). The sixth and seventh columns provide information about any specific method used for subtracting point sources (PS)
and Galactic foregrounds (FG). Note that both the MMF and the ILC methods have a built-in way for subtracting both point sources and diffuse
foregrounds, by treating them as additional noise (of astrophysical origin) correlated across the channels. Note, also, that the study is made only
on clusters at |b| > 20 degrees Galactic latitude for all methods. The eighth column summarizes the main characteritics of each algorithm in terms
of yield at 90% purity and photometric accuracy. Notes. The first column shows the name of the method. The second column indicates when the code is using a prior on the SZ cluster shape. The uperscript a indicate that the detection did not use a shape prior but that the computation of the SZ flux did. The third column gives the
performance in terms of CPU hours needed to complete the analysis. The fourth and fifth column show whether the analysis was made using
all-sky maps or projected patches, their area in square degrees and the number of patches. Methods ILC1 and ILC2 work with full sky maps for
producing an SZ map (by ILC on needlet coefficients) and then work with 504 small patches for cluster detection by matched filtering (ILC1) or
using SExtractor (ILC2). The sixth and seventh columns provide information about any specific method used for subtracting point sources (PS)
and Galactic foregrounds (FG). Note that both the MMF and the ILC methods have a built-in way for subtracting both point sources and diffuse
foregrounds, by treating them as additional noise (of astrophysical origin) correlated across the channels. 3.1.3. MMF3 The MMF3 SZ extraction algorithm is an all-sky extension of the
matched multifrequency filter described in Melin et al. (2006). It has been used for the production of the early SZ cluster sam-
ple (Planck Collaboration 2011b). In the version used for the
SZ challenge, auto and cross power spectra used by the filter do
not rely on prior assumptions about the noise, but are directly
estimated from the data. They are thus adapted to the local in-
strumental noise and astrophysical contamination. 3.2. Bayesian methods 3.2.1. PowellSnakes 3.1.4. MMF4 where H1 is the detection hypothesis, “There is a source” and H0
the null hypothesis “Only background is present” (Jaynes 2003). Applying Bayes theorem to the above formula one gets where H1 is the detection hypothesis, “There is a source” and H0
the null hypothesis “Only background is present” (Jaynes 2003). Applying Bayes theorem to the above formula one gets The spherical matched and scale adaptive filters (Schäfer et al. 2006) are generalisations of the filters proposed by Herranz et al. (2002) for spherical coordinates. Just like their counterparts they
can be derived from an optimisation problem and maximise the
signal to noise ratio while being linear in the signal (matched
filter) and being sensitive to the size of the object. The algo-
rithm interfaces to the common HEALPIX package and treats
the entire celestial sphere in one pass. ρ = Pr(d|H1) Pr(H1)
Pr(d|H0) Pr(H0) = Z1
Z0
Pr(H1)
Pr(H0),
(3) (3) where where where Z =
L(Θ) π(Θ) dDΘ,
(4) The most important drawback is the strong Galactic contam-
ination – the filter was not optimised to deal with a Galactic cut (4) A51, page 5 of 13 A&A 548, A51 (2012) is the evidence, L(Θ) is the likelihood, π(Θ) is the prior and Θ a
vector representing the parameter set. where yi(p) is the observed map in channel i, s(p) is the tem-
plate of interest (here, the SZ map), and ni(p) comprise the
contribution of both all astrophysical foregrounds (CMB, galac-
tic emission, point sources...) and of instrumental noise. This
equation can be recast as: An SZ parameterised template profile of the clusters
s(X, A) ≡ξ τ(a, x −X, y −Y), is assumed known and fairly
representative of the majority of the clusters according to the res-
olution and signal-to-noise ratio of the instrumental setup, where
τ(. . .) is the general shape of the objects (beta or Arnaud et al. profile) and a j a vector which contains the parameters control-
ling the geometry of one specific element (core/scale radius, pa-
rameters of the beta or Arnaud et al. profile). y(p) = as(p) + n(p). (6) (6) y(p) = as(p) + n(p). 3.1.4. MMF4 To first order, linear combinations of the inputs of the form
i wiyi(p) guarantee unit response to the component of inter-
est provided that the constraint
i wiai = 1 is satisfied (there
are, however, restrictions and higher order effects, which are
discussed in detail in the appendix of Delabrouille et al. 2009;
Dick et al. 2010). It can be shown straightforwardly (Eriksen
et al. 2004; Delabrouille & Cardoso 2009) that the linear weights
which minimize the variance of the output map are: The algorithm may be operated on either “Frequentist mode”
where the detection step closely resembles a multi-frequency
multi-scale “matched filter” or “Bayesian mode” where the pos-
terior distributions are computed resorting to a simple “nested
sampling” algorithm (Feroz & Hobson 2008). The acceptance/rejection threshold may be defined either by
using “Decision theory” where the expected loss criterion is
minimised or by imposing a pre-defined contamination ratio. In
the case of a loss criterion, the symmetric criterion – “An unde-
tected cluster is as bad as spurious cluster” – is used. w = atR−1
atR−1a
,
(7) w = atR−1
atR−1a
, (7) where R is the empirical covariance matrix of the observations. What distinguishes the different ILC implementations is essen-
tially the domains over which the above solution is implemented. 3.2.2. BNP This method is described in detail in Diego et al. (2002). It is
based on the maximization of the Bayesian probability of having
an SZ cluster given the multifrequency data with no assumptions
about the shape nor size of the clusters. The method devides the
sky into multiple patches of about 100 sq. degrees and performs
a basic cleaning of the Galactic components (by subtracting the
properly weighted 857 GHz map from the channels of inter-
est for SZ) and point sources (using a Mexican Hat Wavelet). The cleaned maps are combined in the Bayesian estimator and
the output map of Compton parameters is derived. SExtractor is
applied to the map of reconstructed Compton parameter to de-
tect objects above a given threshold and compute their flux. The
thresholds are based on the background or noise estimated by
SExtractor. In order to compute the purity, different signal-to-
noise ratios (ranging from 3 to 10) are used to compute the flux. The method assumes a power spectrum for the cluster population
although this is not critical. 3.3.2. ILC2 The ILC2 approach relies on the same processing for photom-
etry, cluster size, and signal-to-noise estimates as in ILC1, but
in this case the detection of cluster candidates is made using
SExtractor on a Wiener-filtered version of the ILC1 map. The main advantage of the method resides in its robustness
(almost no assumptions) and its ability to reconstruct both ex-
tended and compact clusters. The main limitation is the rel-
atively poor reconstruction of the total flux of the cluster as
compared to matched filters. While the extraction of the ILC map works on full sky maps,
using the needlet framework to perform localized filtering, here
again the detection of cluster candidates, and the estimation
of size and flux, are performed on small patches (obtained by
gnomonic projection). 3.3. Internal linear combination methods The ILC is a simple method for extracting one single compo-
nent of interest out of multifrequency observations. It has been
widely used for CMB estimation on WMAP data (Bennett et al. 2003; Tegmark et al. 2003; Eriksen et al. 2004; Park et al. 2007;
Delabrouille et al. 2009; Kim et al. 2009; Samal et al. 2009). A
general description of the method can be found in Delabrouille
& Cardoso (2009). 3.3.1. ILC1 The needlet ILC method works in two steps. First, an SZ map
is produced by ILC, with a needlet space implementation sim-
ilar to that of Delabrouille et al. (2009). The use of spherical
needlets permits the ILC filter to adapt to local conditions in
both direct (pixel) space and harmonic space. Input maps in-
clude all simulated Planck maps, as well as an external template
of emission at 100 microns (Schlegel et al. 1998), which helps
subtracting dust emission. The cluster catalogue is then obtained
by matched filtering on small patches extracted from the needlet
ILC SZ full-sky map, as described in Melin et al. (2006). yi(p) = ais(p) + ni(p), 3.3.5. ILC5 This algorithm is designed to work on local noisy multichannel
maps in the wavelet domain. The representation of galaxy clus-
ters in an appropriate biorthogonal wavelet basis is expected to
be sparse compared to the contributions of other astrophysical
components. This should ease the subsequent SZ separation us-
ing the ILC method at each wavelet scale. The reverse wavelet
transform is then applied to estimate local SZ-maps. The latter
are convolved using a Gaussian beam of 5 arcmin FWHM to re-
duce the noise prior to cluster detection using SExtractor with a
threshold fixed classically to a multiple of the rms noise (signal-
to-noise ratio ranging from 3 to 6). The brightest IR galaxies and
radiosources are masked to reduce contamination. Finally, mul-
tiple detections due to the overlap of local maps are removed. Multiscale ILC proved to be more efficient than regular ILC to
remove large angular scale contamination on local map simu-
lations. However, the cluster catalogue appears comparatively
more contaminated which may be due to an imperfect clean-
ing of multiple detections. Also, the known point sources were
only masked before detection with SExtractor. Masking the ob-
served sky maps earlier could improve the component separation
using Multiscale ILC. Doing so may prevent a few very bright
pixels biasing the ILC parameter estimation but in turn raises
the question of data interpolation across masked regions. The
other implementations of the ILC do mask point sources before
combining the maps and are thus not subject to this bias. Angular proximity was the only association criterion used
for the matching. Specifically, we matched an extracted cluster
to an input cluster if their separation on the sky θ < θmax = f(θv),
a function of the true angular virial radius of the (input) cluster. The function f(θv) varied over three domains: f(θv) = 5 arcmin
for θv < 5 arcmin; f(θv) = θv for 5 arcmin < θv < 20 arcmin; and
f(θv) = 20 arcmin for θv > 20 arcmin. We first focus on the catalogue completeness and purity, both
of which we define immediately below. We then test the accu-
racy of the recovered flux and size estimates, as well as each
algorithm’s ability to internally evaluate the uncertainties on
these photometric quantities. Since many fewer codes ran on the
Challenge v2, we focus mainly on Challenge v1. 4.1. Catalogue content A useful global diagnostic is the curve of yield versus global pu-
rity for a given catalogue (see e.g. Pires et al. 2006). The former
is simply the total number of clusters detected and the latter we
define as 1 −Γg, where Γg is the global contamination rate: 4. Results We evaluated each extracted catalogue in terms of catalogue
content and photometric recovery based on comparisons be-
tween the extracted catalogues and the simulated input SZ clus- tween the extracted catalogues and the simulated input SZ clus-
ter catalogue. For this purpose, we cut the input catalogue at
Y > 5 × 10−4 arcmin2, well below the theoretical Planck detec-
tion limit (see below), and restrict ourselves to the high latitude
sky at |b| > 20 deg to reduce contamination by galactic fore-
grounds. We then cross-match the candidate cluster in a given
catalogue to a corresponding input cluster. Each match results in
a true detection, while candidates without a match are labeled
as false detections. In a second step, we compare the extracted
properties, namely SZ Compton parameter and size, of the true
detections to the input cluster properties. 3.4. GMCA Generalized morphological component analysis is a blind source
separation method devised for separating sources from instanta-
neous linear mixtures using the model given by: Y = AS + n. The components S are assumed to be sparsely represented (i.e. have a few significant samples in a specific basis) in a so-called
sparse representation Φ (typically wavelets). Assuming that the
components have a sparse representation in the wavelet domain
is equivalent to assuming that most components have a certain
spatial regularity. These components and their spectral signa-
tures are then recovered by minimizing the number of significant
coefficients in Φ: 3.3.3. ILC3 In this method, a filter in the harmonic domain is applied to con-
struct a series of maps that are sensitive to the range of clus-
ter scales. We used a Mexican-hat filter constructed from two
Gaussians, one with 1/4 the width of the other. A list of clus-
ter candidates is compiled using a peak finding algorithm, which
searches for enhanced signal levels in the individual map by fit-
ting cluster model parameter. We employed the β-model profile
with β = 2/3 convolved with the Planck beams. The catalogue
produced then includes as parameters the cluster location, the
flux, and the size estimate. The errors on these parameters are
incorporated as given by the likelihoods of the fit. In this method, a filter in the harmonic domain is applied to con-
struct a series of maps that are sensitive to the range of clus-
ter scales. We used a Mexican-hat filter constructed from two
Gaussians, one with 1/4 the width of the other. A list of clus-
ter candidates is compiled using a peak finding algorithm, which
searches for enhanced signal levels in the individual map by fit-
ting cluster model parameter. We employed the β-model profile
with β = 2/3 convolved with the Planck beams. The catalogue
produced then includes as parameters the cluster location, the
flux, and the size estimate. The errors on these parameters are
incorporated as given by the likelihoods of the fit. The general idea behind the ILC is to form a linear combina-
tion of all available observations which has unit response to the
component of interest, while minimizing the total variance of the
output map. This method assumes that all observations yi(p), for
channel i and pixel p, can be written as the sum of one single
template of interest scaled by some coefficient ai, and of unspec-
ified contaminants which comprise noise and foregrounds, i.e. p
g
y
Additional improvements to the method can be achieved
by using more optimal foreground estimators (but probably (5) yi(p) = ais(p) + ni(p), yi(p) = ais(p) + ni(p), A51, page 6 of 13 A51, page 6 of 13 J.-B. Melin et al.: A comparison of algorithms for the construction of SZ cluster catalogue spectral signature of the free-free component is approximately
known up to a multiplicative constant (power law with fixed
spectral index). with slower convergence). 3.3.4. ILC4 – wavelet denoising; The ILC4 method is a standard ILC in Fourier space performed
on the square patches. It is implemented independently in an-
nuli in wave number (modulus of the Fourier mode) by applying
weights, according to Eq. (7), this time in the Fourier domain. The cluster detection is performed using SExtractor on the re-
constructed SZ map. The flux estimation is performed on the
original multifrequency maps (small patches) using the MMF at
the position of the SExtractor detections. The ILC4 method is a standard ILC in Fourier space performed
on the square patches. It is implemented independently in an-
nuli in wave number (modulus of the Fourier mode) by applying
weights, according to Eq. (7), this time in the Fourier domain. – SExtractor to extract the clusters from the noise-free SZ map,
and finally; – a maximum likehood to get the flux of the detected sources. 3.3.3. ILC3 This is useful especially for the
fainter clusters, or those confused to a high degree by source
contamination. Hence, GMCA furnishes a noisy SZ map in which we want
to detect the SZ clusters. This is done in three steps: 3.3.5. ILC5 We include re-
sults for those codes that did run on Challenge 2 to gauge the in-
fluence of the underlying cluster model used for the simulation. Γg ≡total number of false detections
total number of detections
·
(9) Fewer codes
participated in the Challenge v2 (see text). Fig. 2. For SZ Challenge v2: yield as a function of global purity. The same comment applies concerning the overall yield values; in particular,
the cluster model changed significantly between the versions v1 and v2 of the Challenge resulting in lower overall yields here. Fewer codes
participated in the Challenge v2 (see text). are fully resolved, many have angular sizes comparable to the
effective beam, and this leads to a non-trivial selection function
(White 2003; Melin et al. 2005, 2006). a few square degrees, perform much better than those that em-
ploy a global noise estimate, such as MMF4 and ILC3. For those
methods with local noise estimation, we note that their effec-
tive survey depth appears to anticorrelate with the instrumen-
tal noise, indicating that astrophysical confusion is effectively
removed. This can be seen in Fig. 3, which compares the den-
sity of detected SZ sources (top panel) to the pixel hit count
(bottom panel). The result illustrated with one single method,
ILC2 run on the SZ challenge v1, holds for the other algorithms. The cluster detection limit appears to be primarily modulated by
the instrumental noise at high Galactic latitude, as opposed to
foreground emission. We emphasize that the numerical values of the yields depend
on the cosmological model, on the foreground model and on the
cluster model used in the simulations. They must be considered
with caution because of the inherent modeling uncertainties. As
for the foreground model, the templates used to model Galactic
components in the PSM were chosen so that they are reasonably
representative of the complexity of the diffuse galactic emission. Thanks to many new observations in particular in the IR and
submm domain (Lagache et al. 2007; Viero et al. 2009; Hall
et al. 2010; Amblard et al. 2011; Planck Collaboration 2011d),
the models of point sources have evolved very much between
the beginning of the SZ challenge and the publication of these
results. These updates were not taken into account in the PSM
when the study was performed. Moreover, the cluster model in
challenge v1 was based on the isothermal β-model, while v2
employed a modified NFW pressure profile favoured by X-ray
determinations of the gas pressure (Arnaud et al. 2010) with a
normalization of the Y −M relation lower by ∼15% than in v1. Γg ≡total number of false detections
total number of detections
·
(9) Γg ≡total number of false detections
total number of detections
·
(9) (9) The yield curve is parametrized by the effective detection thresh-
old of the catalogue construction algorithm. It is a global di-
agnostic because it gauges the total content of a catalogue,
rather than its content as a function of flux or other measurable
quantities. Figure 1 compares the yield curves of outputs of all the
algorithms in the SZ Challenge v1, and Fig. 2 those for the
SZ Challenge v2. Increasing the detection threshold moves a
catalogue along its curve to higher purity and lower yield. Algorithms increase in performance towards the upper right-
hand corner, i.e., both high yield and high purity. min{S,S}λ∥SΦT∥+ 1
2∥Y −AS∥2
2
(8) (8) where || . . .||2 is the L2 (Euclidean) norm. In Bobin et al. (2008),
it was shown that sparsity enhances the diversity between the
components thus improving the separation quality. The spec-
tral signatures of CMB and SZ are assumed to be known. The As to be expected, algorithms that locally estimate the noise
(both instrumental and astrophysical), i.e. on local patches of A51, page 7 of 13 A51, page 7 of 13 A&A 548, A51 (2012)
Fig. 1. For SZ Challenge v1: yield as a function of global purity. The right handside panel is a zoom on the high-purity region. Each curve
is parameterized by the detection threshold of the corresponding algorithm. As discussed in the text, the overall value of the yields should be
considered with caution, due to remaining modeling uncertainties (see text). We focus instead on relative yield between algorithms as a measure
of performance. A&A 548, A51 (2012) A&A 548, A51 (2012) Fig. 1. For SZ Challenge v1: yield as a function of global purity. The right handside panel is a zoom on the high-purity region. Each curve
is parameterized by the detection threshold of the corresponding algorithm. As discussed in the text, the overall value of the yields should be
considered with caution, due to remaining modeling uncertainties (see text). We focus instead on relative yield between algorithms as a measure
of performance. Fig. 2. For SZ Challenge v2: yield as a function of global purity. The same comment applies concerning the overall yield values; in particular,
the cluster model changed significantly between the versions v1 and v2 of the Challenge resulting in lower overall yields here. A51, page 8 of 13 Γg ≡total number of false detections
total number of detections
·
(9) Less expected, perhaps, is the fact that all algorithms tend
to miss nearby clusters. These are extended objects, and al-
though they have large total SZ flux, these clusters are “resolved-
out” – missed because of their low surface brightness. This is
an extreme example of resolution effects expected in the case
of SZ detection in relatively low resolution experiments like
Planck. It is not related to the foreground removing efficiency
since the effect can also be mimicked in simulations including
only instrumental white noise. Apart from the few clusters that A51, page 8 of 13 J.-B. Melin et al.: A comparison of algorithms for the construction of SZ cluster catalogues J.-B. Melin et al.: A comparison of algorithms for the construction of SZ cluster catalogues J. B. Melin et al.: A comparison of algorithms for the construction of SZ cluster catalogues
Fig. 3. Illustrated for ILC2 on the SZ Challenge v1, the detection density (top panel) is compared to the pixel hit count for the map at 143 GHz
(bottom panel). The noise in the simulated Planck maps scales as 1/
√
hits. At high Galactic latitudes, the detection density clearly anticorrelates
with map noise. Both maps are smoothed on a scale of 20 degrees, and the Galactic plane (|b| < 20 deg) is masked. Fig. 3. Illustrated for ILC2 on the SZ Challenge v1, the detection density (top panel) is compared to the pixel hit count for the map at 143 GHz
(bottom panel). The noise in the simulated Planck maps scales as 1/
√
hits. At high Galactic latitudes, the detection density clearly anticorrelates
with map noise. Both maps are smoothed on a scale of 20 degrees, and the Galactic plane (|b| < 20 deg) is masked. Finally, σ8 changed from 0.85 in challenge v1 to 0.796 in v2
which strongly influences the total cluster yield. Somewhat deceptively, these yield variations correspond to
only minor differences in detection threshold, as illustrated for
the SZ Challenge v1 in Fig. 4. This figure traces the curves above
which 90% and 10% (lower set and upper set respectively) of
the clusters detected by each method lie in the true Y−true θv
plane. As already mentioned, many clusters are marginally re-
solved (sizes at least comparable to the beam), which means that
detection efficiency depends not only on flux, but also on size6. 6 This was shown on real data in Planck Collaboration (2011b). Γg ≡total number of false detections
total number of detections
·
(9) cosmological interpretation of the counts and hence must be
centred around 10−3 arcmin2, is more contaminated (∼75% on A&A 548, A51 (2012) A&A 548, A51 (2012) A&A 548, A51 (2012) Fig. 4. Selection curves for each algorithm in
the true Y–true θv plane for SZ Challenge v1. The lower set of curves indicate the 90th per-
centiles, i.e., the curve above which lie 90% of
the detected clusters; the upper set corresponds
to the 10th percentile. The colour codes are
as in Fig. 1. We see that the Planck selection
function depends not just on flux, but also on
cluster angular size. Many clusters are at least
marginally resolved by Planck, leading to these
size effects in the selection fuction. The dashed
lines show contours of fixed mass and redshift,
as indicated, while the cloud of points shows
the distribution of the input catalogue (in fact
a subsample of 1 in 4 randomly selected input
clusters). We see that small variations in selec-
tion curves generate significant yield changes. Fig. 5. Positional accuracy of the recovered
clusters illustrated for MMF3 in the SZ chal-
lenge v2. Fig. 5. Positional accuracy of the recovered
clusters illustrated for MMF3 in the SZ chal-
lenge v2. centred around 10−3 arcmin2, is more contaminated (∼75% on
average) in the case of the indirect method than in the case of
direct methods (∼50% on average). cosmological interpretation of the counts and hence must be
properly quantified. cosmological interpretation of the counts and hence must be
properly quantified. We compute for all the methods and in the SZ challenges v1
and v2 the completeness defined as the ratio [true detections
(recovered clusters)/simulated clusters] over bins of true (sim-
ulated) flux Y. We find it varies from 80 to 98% at Ylim =
10−2 arcmin2 for the direct methods (based on frequency maps). The completness is of order of 80% at the same Ylim for the indi-
rect methods (based on detections in SZ maps). We note a slight
increase of the completeness from the SZ Challenge v1 to v2. We
also estimate the contamination of the output catalogues defined
as the ratio [false detections/total detections]. This is evaluated
as as a function of recovered flux. The average contamination
of the output catalogues ranges between 6 and 13% both in the
case of the challenge 1 and 2. Γg ≡total number of false detections
total number of detections
·
(9) However, the purity with respect
to the Y bins differ significantly from method to method. As a
general trend, the lowest Y bin, i.e. the smallet recovered fluxes Γg ≡total number of false detections
total number of detections
·
(9) The more peaked profile actually improves detection ef-
ficiency, while the lower normalization reduces the predicted
yield. Along with the much lower value of σ8, the net result is
that the yields in challenge v2 are noticeably lower than in v1
as seen in Figs. 1 and 2. We thus only discuss, in this study, the
relative yields of the codes as a gauge of performance treating
the absolute value of the yield with caution. The algorithms all have similar curves in this plane. This
means that the differences in yield are due to only small varia-
tions of the selection curve since completeness is expected to be
monotonic. The black points represent a random sample of 1/4
of the input clusters and show where the bulk of the catalogue
lies. These small variations have important consequences for Focusing on the relative merit of the algorithms, we see that
Figs. 1 and 2 display large dispersion in the yield at a given
purity. This reflects of course the intrinsic performance of the
algorithms, but also for the detection methods that share simi-
lar underlying algorithms, e.g. MMF and ILC, the dispersion in
the yield reflects the differences in implementations (e.g. noise
estimation, de-blending, etc.). A51, page 9 of 13 A51, page 9 of 13 A51, page 9 of 13 A&A 548, A51 (2012)
Fig. 4. Selection curves for each algorithm in
the true Y–true θv plane for SZ Challenge v1. The lower set of curves indicate the 90th per-
centiles, i.e., the curve above which lie 90% of
the detected clusters; the upper set corresponds
to the 10th percentile. The colour codes are
as in Fig. 1. We see that the Planck selection
function depends not just on flux, but also on
cluster angular size. Many clusters are at least
marginally resolved by Planck, leading to these
size effects in the selection fuction. The dashed
lines show contours of fixed mass and redshift,
as indicated, while the cloud of points shows
the distribution of the input catalogue (in fact
a subsample of 1 in 4 randomly selected input
clusters). We see that small variations in selec-
tion curves generate significant yield changes. Fig. 5. Positional accuracy of the recovered
clusters illustrated for MMF3 in the SZ chal-
lenge v2. 5. Discussion and conclusion In the present study, we compare different codes and algorithms
to detect SZ galaxy clusters from multi-wavelength experiments
using Plank’s instrumental characteristics. These methods may
be usefully divided into direct methods (four matched-filter ap-
proaches and PowellSnakes) using individual frequency maps,
and indirect methods (five ILC methods, GMCA and BNP) that
first construct an SZ map in which they subsequently search for
clusters. Figure 7 gives the dispersion in the recovered flux σYrec as a
function of true Y (once again, only involving true detections). Here, we see that some codes perform significantly better than
others. Those that adjust an SZ profile to each cluster outper-
form by a large margin those that do not. Photometry based on
SExtractor, for example, fares much worse than the MMF codes. Even among the best performing codes, however, the intrinsic
photometric dispersion is of order 30%. This is important, be-
cause we expect SZ flux to tightly correlate with cluster mass,
with a scatter as low as ∼10% as indicated by both numerical
simulations (Kravtsov et al. 2006) and recent X-ray data (Arnaud
et al. 2007; Nagai et al. 2007). The SZ flux hence should offer
a good mass proxy. What we see from this figure, however, is
that the observational scatter will dominate the intrinsic scatter
of this mass proxy and needs to be properly accounted for in the
cosmological analyses. As already emphasized, the global yield values of all meth-
ods must be considered with caution because of inherent model-
ing uncertainties of the sky simulations and cluster models used,
and to the underlying cosmological model. Therefore, we focus
on relative yields as a gauge of performance of the algorithms. It
is worth noting that results of a direct or indirect method signif-
icantly vary (within factors of as much as three) with the details
of their implementation, with clear impact on the survey yield as
demonstrated in Figs. 1 and 2. Using the PSM simulations and
including th noise as described in 2.1, we would expect of order
of 1000–2000 clusters at a purity of ∼90% with |b| > 20 deg. This number depends on the extraction method used and may
vary with a more detailed modeling of the sky. The cluster
yield can be increased by accepting a higher contamination rate
and calling for extensive follow-up to eliminate false detections
a posteriori. 4.2. Photometric and astrometric recovery Cluster characterization is a separate issue from detection. It in-
volves determination of angular positions as well as photometry. Since Planck will marginally resolve many clusters, photometry
here means both flux Yrec and characteristic size measurements. Moreover, each method should provide an estimate of the errors
on these quantities for each object in the catalogue. We illustrate in Fig. 5, a scatter diagram of positional off-
set for MMF3 as a function of true cluster size, θv. On av-
erage, all the algorithms perform similarly and recover clus-
ter position to ∼2 arcmin with a large scatter. In addition, we
see that it is more difficult to accurately determine the position A51, page 10 of 13 J.-B. Melin et al.: A comparison of algorithms for the construction of SZ cluster catalogues J. B. Melin et al.: A comparison of algorithms for the construction of SZ cluster catalogues
Fig. 6. Flux recovery bias. The figure shows for
a subset of methods the average recovered Yrec,
normalized to the true input Y, as a function
of Y. At the bright end, most codes extract an
unbiased estimate of cluster flux, while the ex-
pected Malmquist bias appears at the faint-end,
just below Y ∼2 × 10−3 arcmin2. Fig. 7. Flux recovery uncertainty for the subset
of methods given in Fig. 6. The Figure shows
the dispersion in measured flux about the true
input Y flux as a function of the true input
flux. The best algorithms are affected by ∼30%
dispersion. Fig. 7. Flux recovery uncertainty for the subset
of methods given in Fig. 6. The Figure shows
the dispersion in measured flux about the true
input Y flux as a function of the true input
flux. The best algorithms are affected by ∼30%
dispersion. Fig. 7. Flux recovery uncertainty for the subset
of methods given in Fig. 6. The Figure shows
the dispersion in measured flux about the true
input Y flux as a function of the true input
flux. The best algorithms are affected by ∼30%
dispersion. of intrinsically extended clusters, as shown by the fact that the
cloud of points is elongated and inclined. this issue arises specifically for Planck-like resolution because a
large number of clusters are only marginally resolved. Imposing
the cluster size, for example from external data, such as X-ray
or optical observations or higher resolution SZ measurements,
would significantly reduce the observational scatter. 4.2. Photometric and astrometric recovery Concerning photometry, we show in Figs. 6 and 7 the mean
recovered SZ flux, Yrec, normalized to the true (simulated) flux,
Y, as a function of the latter. Only true detections are used in this
comparison. We illustrate our results for a subset of methods
namely MMF1, MMF2, MMF3, ILC1, ILC2, ILC3. Methods
that filter the maps such as ILC4 and GMCA, or that use the
SExtractor photometry such as BNP and ILC5 exhibit significant
bias in flux recovery at the bright end. At the faint end, we see the
appearance of Malmquist bias as an upturn in the measured flux. The importance of this bias varies from algorithm to algorithm. Fig. 6. Flux recovery bias. The figure shows for
a subset of methods the average recovered Yrec,
normalized to the true input Y, as a function
of Y. At the bright end, most codes extract an
unbiased estimate of cluster flux, while the ex-
pected Malmquist bias appears at the faint-end,
just below Y ∼2 × 10−3 arcmin2. 5. Discussion and conclusion We have attributed the origin of the photometric scatter to
difficulty in determining cluster size. Although methods adjust-
ing a profile to the SZ are able to estimate the size of many
clusters, they do so with significant dispersion. Furthermore, A51, page 11 of 13 A51, page 11 of 13 A&A 548, A51 (2012) The indirect methods seem to offer greater opportunity for
optimization with a larger number of tuning parameters. They
are also less model dependent for the SZ map construction
and the cluster detection. Although, they can be coupled with
matched filters for the SZ flux measurement. In turn, the direct
methods are linear, easy to implement and robust. One of their
advantages relies in the fact that they can be optimized to de-
tect objects of a given shape (SZ profile) and and a given spec-
tral energy distribution (SED; SZ spectrum). This characteris-
tic of the direct method is particularly important for Planck-like
multi-frequency surveys with moderate resolution. Indeed, due
to lack of resolution, spurious sources (galactic features or point
sources) may be detected by the spatial filter and in that case the
frequency coverage and the spectral matching is the best strat-
egy to monitor the false detections. Due to their robustness and
easy implementation direct methods are more adapted to run in
pipelines7. The situation is quite different for high resolution,
arcminute-scale, SZ experiments such as ACT and SPT where
the filtering of one unique low frequency map (where the SZ sig-
nal is negative) is sufficiently efficient to unable cluster detec-
tion. In these cases though, extended clusters are not well recov-
ered as they suffer more from the CMB contamination and thus
from the filtering. marginally resolved by Planck: large enough that their angular
extent matters, but small enough that we have difficulty fixing
their true size. One way of reducing the photometric error is
thus using external constraints on cluster size. One again an-
cillary data from RASS or optical cluster catalogues will help
in this regard, at least at low redshift (z < 0.3−0.5); at higher
redshift, we will rely on follow-up observations if we want to
reduce photometric uncertainties. 5. Discussion and conclusion The comparison of an ensemble of cluster extraction meth-
ods in the case of a multi-frequency moderate resolution experi-
ment shows that the optimization of the cluster detection in terms
of yield and purity, but also in terms of positional accuracy and
photometry, is very sensitive to the implementation of the code. The global or local treatment of the noise estimate or the clean-
ing from point sources are the two main causes of difference. However and most importantly, the use of as realistic as possi-
ble SZ profile (as opposed to model independent profile) to filter
out the signal or to measure the fluxes is a key aspect of cluster
detection techniques in our context. In that respect, using exter-
nal information from SZ observations or from other wavelength
will significantly help in improving the measurement of the clus-
ter properties and in turn optimize the catalogue yields and their
selection function. g
The comparison of different codes and cluster detection
methods exhibits selection effects and catalogue uncertainties,
neither of which depend, for example, on the actual cluster
physics model. This is shown in the selection curves Fig. 4. In
a Planck-like case, with moderate resolution, clusters do not ap-
pear as point sources, but many are resolved or have sizes com-
parable to the effective SZ beam. In view of this, the use of an
adapted spatial filter to optimally model the SZ profile provides
a significant improvement in the detection yield and in the pho-
tometry. Clusters being partially resolved leads to non-trivial se-
lection criteria that depend both on flux and true angular size, as
demonstrated by the fact that the curves in Fig. 4 are not horizon-
tal lines. In that respect, the use of X-ray information from the
ROSAT All-Sky Survey (RASS) cluster catalogues (Böhringer
et al. 2000, 2004; Piffaretti et al. 2011) or from optical cata-
logues, e.g. in the SDSS area (Koester et al. 2007), will be of par-
ticular value as they will give us a handle on both flux and size
of the clusters detected by Planck and, even more importantly,
understand better the completeness in studying those which are
missed (Chamballu et al. 2012). As already stated, the exact po-
sition of each selection curve in Fig. 4 depends on the algorithm
and small variations in the position of this curve produce signif-
icant changes in catalogue yield. Acknowledgements. The authors acknowledge the use of the Planck Sky
Model (PSM), developed by the Component Separation Working Group (WG2)
of the Planck Collaboration. We acknowledge also the use of the HEALPix pack-
age (Górski et al. 2005). We further thank M. White and S. White for helpful
comments and suggestions. 7 MMF1, MM2, MMF3 and PwS were all implemented in Planck’s
pipelines. Furthermore, MMF3 was used to extract the Planck clusters
published in the ESZ sample. 5. Discussion and conclusion Most of the differences bew-
teen catalogues occur at small flux and size, where the bulk of
the cluster population resides. These objects are for the major-
ity low mass, intermediate redshift clusters. Moreover, detection
becomes progressively less efficient for large objects; this is in-
trinsic and hence shared by all algorithms. References Albrecht, A., Bernstein, G., Cahn, R., et al. 2006 [arXiv:astro-ph/0609591]
Albrecht, A., Amendola, L., & Bernstein, G. 2009 [arXiv:0901.0721]
Amblard, A., Cooray, A., Serra, P., et al. 2011, Nature, 470, 510
Arnaud, M., Pointecouteau, E., & Pratt, G. W. 2007, A&A, 474, L37
Arnaud M Pratt G W Piffaretti R et al 2010 A&A 517 A92 Albrecht, A., Bernstein, G., Cahn, R., et al. 2006 [arXiv:astro-ph/0609591]
Albrecht, A., Amendola, L., & Bernstein, G. 2009 [arXiv:0901.0721] Amblard, A., Cooray, A., Serra, P., et al. 2011, Nature, 470, 510
A
d M P i
E & P
G W 2007 A&A 474 L y
Arnaud, M., Pointecouteau, E., & Pratt, G. W. 2007, A&A, 474, L37
Arnaud M Pratt G W Piffaretti R et al 2010 A&A 517 A92 Bartlett, J. G. 2002, in Tracing Cosmic Evolution with Galaxy Clusters, eds. S. Borgani, M. Mezzetti, & R. Valdarnini, ASP Conf. Ser., 268, 101 Bartlett, J. G. 2002, in Tracing Cosmic Evolution with Galaxy Clusters, eds. S B
i M M
i & R V ld
i i ASP C
f S
268 101 S. Borgani, M. Mezzetti, & R. Valdarnini, ASP Conf. Ser., 268, 10 g
Bennett, C. L., Hill, R. S., Hinshaw, G., et al. 2003, ApJS, 148, 97 Bertin, E., & Arnouts, S. 1996, A&AS, 117, 393 Birkinshaw, M. 1999, Phys. Rep., 310, 97 Bobin, J., Moudden, Y., Starck, J., Fadili, J., & Aghanim, N. 2008, Stat. Method.,
5, 307 Böhringer, H., Voges, W., Huchra, J. P., et al. 2000, ApJS, 129, 435
Böhringer, H., Schuecker, P., Guzzo, L., et al. 2004, A&A, 425, 367 g ,
,
g
,
,
,
,
,
p
,
,
Böhringer, H., Schuecker, P., Guzzo, L., et al. 2004, A&A, 425, 367 g
Bonaldi, A., Tormen, G., Dolag, K., & Moscardini, L. 2007, MNRAS, 378, 1248 Carlstrom, J. E., Holder, G. P., & Reese, E. D. 2002, ARA&A, 40, Carvalho, P., Rocha, G., & Hobson, M. P. 2009, MNRAS, 393, 681 Chamballu, A., Bartlett, J. G., & Melin, J. 2012, A&A, 544, A40 Colafrancesco, S., Mazzotta, P., Rephaeli, Y., & Vittorio, N. 1997, da Silva, A. C., Kay, S. T., Liddle, A. R., & Thomas, P. A. 2004, MNRAS, 348,
1401 de Zotti, G., Ricci, R., Mesa, D., et al. 2005, A&A, 431, 893 de Zotti, G., Ricci, R., Mesa, D., et al. 2005, A&A, 431, 893
Delabrouille, J., & Cardoso, J. References 2009, in Lecture Notes in Physics 665, eds. V. J. Martinez, E. Saar, E. M. Gonzales, & M. J. Pons-Borderia
(Berlin:
Springer Verlag), 159 Concerning individual cluster measurements we find that the
astrometry is recovered to ∼2 arcmin on average and photometry
to ∼30% for the best-behaving algorithms. The positional error
is not a problem for X-ray/optical follow-ups because Planck is
expected to detect massive clusters which will be easy to find
in the XMM/Chandra/1 to 4 m class optical telescope fields of
view. The photometric error in our Planck-like case is dominated
by the difficulty in accurately determining the cluster extent/size. This is another consequence of the fact that many clusters are p
g
g
Delabrouille, J., Cardoso, J., Le Jeune, M., et al. 2009, A&A, 493, 835 Delabrouille, J., Betoule, M., Melin, J.-B., et al. 2012, A&A, submitted
[arXiv:1207.3675] Dick, J., Remazeilles, M., & Delabrouille, J. 2010, MNRAS, 401, 1602 Dickinson, C., Davies, R. D., & Davis, R. J. 2003, MNRAS, 341, 369 Diego, J. M., Vielva, P., Martínez-González, E., Silk, J., & Sanz, J. L. 2002,
MNRAS, 336, 1351 Draine, B. T., & Lazarian, A. 1998, ApJ, 494, L19 Draine, B. T., & Lazarian, A. 1998, ApJ, 494, L19 Dunkley, J., Komatsu, E., Nolta, M. R., et al. 2009, ApJS, 180, 306 Eke, V. R., Navarro, J. F., & Frenk, C. S. 1998, ApJ, 503, 569 p
Eriksen, H. K., Banday, A. J., Górski, K. M., & Lilje, P. B. 2004, ApJ, 612, 633 Feroz, F., & Hobson, M. P. 2008, MNRAS, 384, 449 Finkbeiner, D. P., Davis, M., & Schlegel, D. J. 1999, ApJ, 524, 8 Fomalont, E. B., Windhorst, R. A., Kristian, J. A., & Kellerman, K. I. 1991, AJ,
102, 1258 A51, page 12 of 13 aynes, E. T. 2003, Probability Th
(Cambridge University Press) Samal, P. K., Saha, R., Delabrouille, J., et al. 2010, ApJ, 714, 840 Jenkins, A., Frenk, C. S., White, S. D. M., et al. 2001, MNRAS, 321, 372 Jenkins, A., Frenk, C. S., White, S. D. M., et al. 2001, MNRAS, 321, 372
Kim J Naselsky P & Christensen P R 2009 Phys Rev D 79 023003 Schäfer, B. M., Pfrommer, C., Hell, R. M., & Bartelmann, M. 2006, MNRAS,
370, 1713 Jenkins, A., Frenk, C. S., White, S. D. M., et al. 2001, MNRAS, 321, 372
Kim, J., Naselsky, P., & Christensen, P. R. 2009, Phys. Rev. D, 79, 023003 Schlegel, D. J., Finkbeiner, D. P., & Davis, M. 1998, ApJ, 500, 525 Koester, B. P., McKay, T. A., Annis, J., et al. 2007, ApJ, 660, 239 ,
,
y,
,
,
,
,
p ,
,
Kravtsov, A. V., Vikhlinin, A., & Nagai, D. 2006, ApJ, 650, 128 g
p
Schuecker, P., Böhringer, H., Collins, C. A., & Guzzo, L. 2003, A&A, 398, 867 Kravtsov, A. V., Vikhlinin, A., & Nagai, D. 2006, ApJ, 650, 128 Serjeant, S., & Harrison, D. 2005, MNRAS, 356, 192 Lagache, G., Bavouzet, N., Fernandez-Conde, N., et al. 2007, ApJ, 665, L89 Shaw, L. D., Holder, G. P., & Bode, P. 2008, ApJ, 686, 206 Leach, S. M., Cardoso, J., Baccigalupi, C., et al. 2008, A&A, 491, 5 eac , S. ., Ca doso, J.,
acc ga up , C., et a . 008,
& , 9 , 597
Melin, J.-B., Bartlett, J. G., & Delabrouille, J. 2005, A&A, 429, 417
M li
J B B tl tt J G & D l b
ill
J 2006 A&A 459 341 Melin, J.-B., Bartlett, J. G., & Delabrouille, J. 2005, A&A, 429, 417 Sheth, R. K., & Tormen, G. 1999, MNRAS, 308, 119 Sheth, R. K., & Tormen, G. 1999, MNRAS, 308, 119 Melin, J.-B., Bartlett, J. G., & Delabrouille, J. 2006, A&A, 459, 341 Sunyaev, R. A., & Zeldovich, Y. B. 1970, Comm. Astrophys. Space Phys., 2, 66 Sunyaev, R. A., & Zeldovich, Y. B. 1970, Comm. Ast Miville-Deschênes, M. 2009 [arXiv:0905.3094] Sunyaev, R. A., & Zeldovich, Y. B. 1972, Comm. Astrophys. Space Phys., 4,
173 Miville-Deschênes, M., Lagache, G., Boulanger, F., & Puget, J. 2007, A&A, 469,
595 Tegmark, M., de Oliveira-Costa, A., & Hamilton, A. J. 2003, Phys. Rev. J.-B. Melin et al.: A comparison of algorithms for the construction of SZ cluster catalogues Gaustad, J. E., McCullough, P. R., Rosing, W., & Van Buren, D. 2001, PASP,
113, 1326 Planck Collaboration 2011a, A&A, 536, A6 Planck Collaboration 2011b, A&A, 536, A8 ,
Górski, K. M., Hivon, E., Banday, A. J., et al. 2005, ApJ, 622, 759 Planck Collaboration 2011c, A&A, 536, A11 Planck Collaboration 2011d, A&A, 536, A18 Granato, G. L., De Zotti, G., Silva, L., Bressan, A., & Danese, L. 2004, ApJ,
600, 580 Planck Collaboration 2011e, A&A, 536, A10 Haffner, L. M., Reynolds, R. J., Tufte, S. L., et al. 2003, ApJS, 149, 405 Haffner, L. M., Reynolds, R. J., Tufte, S. L., et al. 2003, ApJS, Planck Collaboration 2011f, A&A, 536, A9 Planck Collaboration 2011g, A&A, 536, A12 ll, N. R., Keisler, R., Knox, L., et al. 2010, ApJ, 718, 632 Haslam, C. G. T., Salter, C. J., Stoffel, H., & Wilson, W. E. 1982, A&AS, 47, 1
Herranz D Sanz J L Hobson M P et al 2002 MNRAS 336 1057 Reinecke, M., Dolag, K., Hell, R., Bartelmann, M., & Enßlin, T. A. 2006, A&A,
445, 373 Haslam, C. G. T., Salter, C. J., Stoffel, H., & Wilson, W. E. 1982, A&AS, 47, 1
Herranz, D., Sanz, J. L., Hobson, M. P., et al. 2002, MNRAS, 336, 1057 Herranz, D., Sanz, J. L., Hobson, M. P., et al. 2002, MNRAS, 336, 1057 Rosati, P., Borgani, S., & Norman, C. 2002, ARA&A, 40, 539 Jaynes, E. T. 2003, Probability Theory: the logic of science, ed. G. L. Bretthorst
(Cambridge University Press) aynes, E. T. 2003, Probability Th
(Cambridge University Press) D, 68,
123523 Toffolatti, L., Argueso Gomez, F., de Zotti, G., et al. 1998, MNRAS, 297, 117 Motl, P. M., Hallman, E. J., Burns, J. O., & Norman, M. L. 2005, ApJ, 623, L63 Motl, P. M., Hallman, E. J., Burns, J. O., & Norman, M. L. 2005, ApJ, 623, L63
Nagai, D. 2006, ApJ, 650, 538 ,
,
,
,
,
,
,
,
p ,
,
Nagai, D. 2006, ApJ, 650, 538 Nagai, D. 2006, ApJ, 650, 538 Truemper, J. 1992, QJRAS, 33, 165 Truemper, J. 1992, QJRAS, 33, 165 g
p
Nagai, D., Kravtsov, A. V., & Vikhlinin, A. 2007, ApJ, 668, 1 Viero, M. P., Ade, P. A. R., Bock, J. J., et al. 2009, ApJ, 707, 176 Park, C., Park, C., & Gott, III, J. R. 2007, ApJ, 660, 959 Vikhlinin, A., Kravtsov, A. V., Burenin, R. A., et al. 2009, ApJ, 692, 1060
Voit, G. M. 2005, Rev. Modern Phys., 77, 207 Piffaretti, R., Arnaud, M., Pratt, G. W., Pointecouteau, E., & Melin, J.-B. 2011,
A&A, 534, A109 y
Wakker, B. P., York, D. G., Wilhelm, R., et al. 2008, ApJ, 672, 298
White, M. 2003, ApJ, 597, 650 y
Wakker, B. P., York, D. G., Wilhelm, R., et al. 2008, ApJ, 672, 298 Pires, S., Juin, J. B., Yvon, D., et al. 2006, A&A, 455, 741 A51, page 13 of 13
|
https://openalex.org/W4292959968
|
https://hal-pasteur.archives-ouvertes.fr/pasteur-03854881/file/Bourdon-2022-stuart_%20an%20R%20package%20for%20the%20cura.pdf
|
English
| null |
stuart: an R package for the curation of SNP genotypes from experimental crosses
|
G3
| 2,022
|
cc-by
| 6,485
|
To cite this version: Marie Bourdon, Xavier Montagutelli. stuart: an R package for the curation of SNP genotypes from
experimental crosses. G3, 2022, 12, pp.jkac219. 10.1093/g3journal/jkac219. pasteur-03854881 Distributed under a Creative Commons Attribution 4.0 International License Introduction Genetic mapping of Mendelian or quantitative traits in inbred
strains is classically achieved in 2-generation crosses such as
intercrosses (F2) and backcrosses (N2), in which the inheritance
of the trait is compared with the genotypes at multiple genetic
markers encompassing the genome map. Variations of a quanti-
tative trait are controlled by one or more quantitative trait loci
(QTLs). A QTL is defined as a marker at which individuals carry-
ing different genotypes show different average trait values. QTL
mapping searches for QTLs by testing the association between
trait values and genotypes at markers spanning the genome
map. The statistical significance of the association is expressed
as logarithm of the odds (LOD) score which is calculated for each
genotyped marker and, at intermediates positions, for pseudo-
markers created by interval mapping, generating an LOD score
curve (Broman 2001). The curve peaks at regions potentially asso-
ciated with the trait. These peaks are called QTLs if they reach
predefined statistical thresholds established either from general
statistical models (Lander and Kruglyak 1995) or by permutation
tests performed on the cross data. For each permutation, pheno-
types are shuffled between individuals to break real associations,
and LOD scores are calculated to identify peaks, which are all
false positives. The distribution of the peak LOD scores over a
large
number
(>1,000)
of
permutations
provides
statistical
thresholds: if a LOD score of 3.8 or higher is observed in 5% of the
permutations, this value will be taken as the P ¼ 0.05 threshold
(Doerge and Churchill 1996). QTL mapping on F2s and N2s can be
conducted with R packages such as R/qtl (Broman et al. 2003) and
R/qtl2(Broman et al. 2019). Several tools exist for quality control of SNP genotyping arrays,
including Illumina’s GenomeStudio. R packages such as argyle
(Morgan 2015) analyze hybridization intensity signals from MUGA
arrays. The simple genetic structure of 2-generation crosses pro-
vides specific and efficient means for identifying spurious genotyp-
ing data, such as consistency with parental genotypes and expected
Mendelian proportions. The R/qtl package includes functions to
build genetic maps and check for genotype consistency (https://rqtl. org/tutorials/geneticmaps.pdf). However, this control is performed
once genotypes have been imported and involves multiple steps of
manual curation. Received: July 5, 2022. Accepted: August 19, 2022
V
C The Author(s) 2022. Published by Oxford University Press on behalf of Genetics Society of America.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which
permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Genetic mapping in 2-generation crosses requires genotyping, usually performed with single nucleotide polymorphism markers arrays which
provide high-density genetic information. However, genetic analysis on raw genotypes can lead to spurious or unreliable results due to
defective single nucleotide polymorphism assays or wrong genotype interpretation. Here, we introduce stuart, an open-source R package,
which analyzes raw genotyping data to filter single nucleotide polymorphism markers based on informativeness, Mendelian inheritance
pattern, and consistency with parental genotypes. The functions of this package provide a curation pipeline and formatting adequate for
genetic analysis with the R/qtl package. stuart is available with detailed documentation from https://gitlab.pasteur.fr/mouselab/stuart/. Keywords: R package; genetic analysis; SNP genotypes high frequency, low cost, and high-throughput analysis using
various genotyping platforms. In mice, several generations of
Mouse Universal Genotyping Arrays (MUGA) have been devel-
oped, the most recent being GigaMUGA (143k SNPs; Morgan et al. 2015) and MiniMUGA (10.8k SNPs; Sigmon et al. 2020). GigaMUGA
provides high-density coverage for the fine characterization of in-
bred strains or outbred populations such as the Diversity Outbred
(Svenson et al. 2012), while the modest number of SNPs in
MiniMUGA is largely sufficient to genotype intercrossed or back-
crossed individuals. However, SNP reliability is affected by the
performance of genotyping platforms and polymorphism be-
tween and within inbred strains. Spurious or unreliable mapping
outputs can result from defective SNP assays or wrong genotype
interpretations. Therefore, raw data obtained from genotyping
services must be curated before performing genetic analyses. G3, 2022, jkac219
https://doi.org/10.1093/g3journal/jkac219
Advance Access Publication Date: 24 August 2022
Software and Data Resources Downloaded from https://academic.oup.com/g3journal/advance-article/doi/10.1093/g3journal/jkac219/6674511 by Institut Pasteur - CeRIS user on 28 September 2022 stuart: an R package for the curation of SNP genotypes
from experimental crosses Marie Bourdon
, Xavier Montagutelli
*
Mouse Genetics Laboratory, Institut Pasteur, Universite´ Paris Cite´, F-75015 Paris, France *Corresponding author: Mouse Genetics Laboratory, Institut Pasteur, Universite´ Paris Cite´ , F-75015 Paris, France. Email: xavier.montagutelli@pasteur.fr *Corresponding author: Mouse Genetics Laboratory, Institut Pasteur, Universite´ Paris Cite´ , F-75015 Paris, France. Email: xav HAL Id: pasteur-03854881
https://pasteur.hal.science/pasteur-03854881v1
Submitted on 16 Nov 2022 L’archive ouverte pluridisciplinaire HAL, est
destinée au dépôt et à la diffusion de documents
scientifiques de niveau recherche, publiés ou non,
émanant des établissements d’enseignement et de
recherche français ou étrangers, des laboratoires
publics ou privés. HAL is a multi-disciplinary open access
archive for the deposit and dissemination of sci-
entific research documents, whether they are pub-
lished or not. The documents may come from
teaching and research institutions in France or
abroad, or from public or private research centers. Distributed under a Creative Commons Attribution 4.0 International License Materials and methods stuart is a tidyverse (Wickham et al. 2019) based R package requir-
ing R version 3.5.0 or later. Its open source is available on Institut
Pasteur’s GitLab: https://gitlab.pasteur.fr/mouselab/stuart/ and
can be installed with devtools (Wickham et al. 2021). stuart’s vi-
gnette provides detailed descriptions of data import and of each
function.stuart imports SNP allele calls from MUGA Illumina
platform or other sources using the same file format. The central
object of stuart is the marker table which summarizes for each
marker, the alleles found in the population, the number of indi-
viduals of each genotype and the exclusion status resulting from
the curation steps. stuart exports curated data to an R/qtl com-
patible format. The SNP annotation file used was downloaded
from https://raw.githubusercontent.com/kbroman/MUGAarrays/
master/UWisc/mini_uwisc_v2.csv
(last
accessed
August
29,
2022). g
y
p
These 2 consequences of genotyping inconsistencies are illus-
trated in Fig. 1c and Supplementary Fig. 3, a and b which were
obtained using the R/qtl scanone() function on a quantitative
trait from the uncurated F2 datasets. For dataset 1, the P ¼ 0.05
significance threshold was estimated at 19.4 (Fig. 1c), while it
usually ranges between 3.3 and 4.3 depending on the inheritance
model for crosses of this type and size (Lander and Kruglyak
1995). Several peaks were detected although none reached P
¼ 0.05 significance. Moreover, their narrow profile was highly un-
expected in F2 crosses. Indeed, these peaks involved only 1–3
markers, and the LOD score curve felt abruptly between these
and adjacent markers on both sides (Fig. 1d), while genetic link-
age between closely linked markers should result in progressive
decrease of the LOD score curve on both sides of a peak (Gue´net
et al. 2015). Among the 3 datasets, we identified 4 narrow peaks
reaching suggestive significance level (P < 0.63): 2 were located at
a marker with non-Mendelian allelic proportions and 2 were
located at 1–3 pseudomarkers adjacent to a marker with non-
Mendelian proportions (Supplementary Fig. 3, c, d and e, f,
respectively). We identified 5 other narrow peaks (LOD score be-
tween 6.72 and 10.03) out of which 4 resulted from the same
situations as above and one was located on a pseudomaker and a
marker with non-Mendelian proportions. Three datasets were used to test the package. This article
presents the results from 176 (CC001/Unc X C57BL/6J-Ifnar1 KO)
F2 mice (dataset 1). Introduction To provide a more automated process of data
curation before genetic analysis, we have developed stuart, an R
package that implements a pipeline for automatic filtering and
curation of SNP genotyping data from 2-generation crosses based
on simple rules. This package formats raw SNP allele calls from
Illumina files into genotypes ready for importation in R/qtl. Using 3
intercross datasets, we illustrate the consequences of inconsistent With genome sequencing, single nucleotide polymorphisms
(SNPs) have become the standard across species for their very 2
|
G3, 2022, Vol. 00, No. 0 distribution of the ratio between the calculated and the known
genetic
distances
between
adjacent
markers
(Fig. 1b
and
Supplementary Fig. 2, c and d; to avoid exaggerated ratios, we
considered only markers with a known distance of 1 cM or more). This analysis revealed 2 groups of markers. In dataset 1, for 43%
% of them, the ratio was below 5 and followed a Gaussian distri-
bution with mean ¼ 1.31 and SD ¼ 0.77. The other markers (57%)
showed a ratio between 5 and 981.87 (Fig. 1b) which necessarily
results from incorrect genotypes, as only a few individuals should
show recombination between adjacent markers. On chromosome
1, while the known marker positions spanned 100 cM, the cu-
mulated genetic distance estimated from observed RF was
40,000 cM. As QTL mapping relies on coherent genotypes at a
series of markers encompassing a genetic interval, problematic
genotypes at a given marker will perturb the analysis and, in
some cases, may result in peaks of the LOD score curve in the ab-
sence of true association (Cheung et al. 2014). Such false positives
increase significance thresholds calculated by data permutation. genotypes on the estimated marker map and QTL mapping, and
how the curation achieved by each function in stuart leads to trust-
able results. Downloaded from https://academic.oup.com/g3journal/advance-article/doi/10.1093/g3journal/jkac219/6674511 by Institut Pasteur - CeRIS user on 28 September 2022 Results and discussion
Consequences of inconsistent genotypes SNP data delivered by the Illumina platform are base alleles that
need to be translated into genotypes for genetic analysis. From
our experience on multiple 2-generation crosses, we identified
several types of genotype inconsistencies that were responsible
for distorted marker maps and spurious QTL mapping results. Recombination fraction (RF), which measures the genetic dis-
tance between 2 markers, is estimated in a cross by analyzing the
proportion of recombinants between adjacent markers in all indi-
viduals. The map of markers calculated from the cross data
should be consistent with their known positions. The R/qtl est.-
map() and plotMap() functions produce a graphical comparison
of the 2 maps (Fig. 1a and Supplementary Fig. 2, a and b). For
each chromosome, the known position of each marker provided
in the annotation file (left) is connected with the estimated posi-
tion (right) based on observed RF. With minimally curated geno-
types (exclusion of nonpolymorphic markers and markers with
over 50% missing genotypes), large RF was found in many instan-
ces between closely linked markers, resulting in fan-like patterns. To
further
describe
these
distortions,
we
computed
the Results and discussion Inconsistent marker maps may also originate from the wrong
assignment of markers to their chromosome and position pro-
vided to the mapping program. Indeed, R/qtl developer K. Broman identified errors in MUGA arrays annotation files affect-
ing marker positions, probe sequences mapping to several loca-
tions, and unmappable markers. We recommend using K. Broman’s corrected annotation files available on GitHub. The
conversion of SNP alleles (A, C, T, G) observed in second-
generation individuals (SGIs) to genotypes encoded according to
the parental alleles may also create genotype errors. Reference
SNP alleles established for many mouse strains may be used to
infer the SGI genotypes. However, we recommend genotyping
individuals of the parental strains used in the cross since they
could differ from the reference panel. In our example dataset, the
2 parental strains used in the cross showed allelic differences
with their reference panel counterpart at 200 markers. Materials and methods The analysis of 2 other data sets, 94 (C57BL/
6J-Ifnar1 KO X 129S2/SvPas-Ifnar1 KO) F2 mice (dataset 2) and 89
(C57BL/6NCrl X CC021/Unc) F2 mice (dataset 3) is presented as
Supplementary data. Quantitative traits were studied in the 3
F2s. Phenotype distributions are presented in Supplementary
Fig. 1. Genotyping was performed by Neogen (Auchincruive,
Scotland) with MiniMUGA on DNA prepared from tail biopsies us-
ing standard phenol-chloroform extraction. Genotype call rate
was 0.927, 0.931, and 0.948 for dataset 1, dataset 2, and dataset 3,
respectively. QTL mapping was performed using R/qtl. Statistical
significance of phenotype–genotype association was computed
by data permutation (Doerge and Churchill 1996), which provides
genome-wide thresholds accounting for multiple testing. The fol-
lowing thresholds were used, as commonly accepted (Members
of the Complex Trait Consortium 2003): P ¼ 0.05 for significant as-
sociation, P ¼ 0.1 and P ¼ 0.63 for the suggestive association. All
figures were designed with ggplot2 (Wickham 2016) or R/qtl. Data control and curation performed in stuart The estimated map is considerably expanded because of multiple genotype inconsistencies. b) Distribution of
the ratio between estimated and known distances between adjacent markers. Markers with known and calculated distances below 1 cM were removed
as they may lead to extremely small or large ratios. The expansion of the estimated map leads to a distribution tail of high ratios. The y-axis is in
logarithmic scale. Fifty-seven percent of markers have a ratio above 5 (dashed line). c) Output of the scanone function of R/qtl showing the
identification of narrow LOD score peaks. Genome-wide significance thresholds computed by data permutation are shown as plain (P ¼ 0.05), dotted
(P ¼ 0.1), and dashed (P ¼ 0.63) lines. d) Magnification of the scanone plot restricted to chromosome 13 (peak p2). The LOD score peak is located on one
marker (red tick) distant by 1.728 and 1.24 cM from the proximal and distal markers, respectively, on the known marker map, but by 1,001.582 and
1,001.506 cM based on calculated RF. performed by each function, and the number of markers of data-
set 1 retained after each step. SNP markers. If parental and SGI data were produced on different
versions, the marker lists must be compared to retain only com-
mon SNPs. This is achieved by the mark_match() function. Data control and curation performed in stuart Although each of stuart’s functions can be called independently,
we present a logical analysis workflow appropriate for 2-genera-
tion crosses. Table 1 summarizes the data curation and filtering M. Bourdon and X. Montagutelli
|
3 M. Bourdon and X. Montagutelli
|
3 (a)
(b)
(c)
(d)
Fig. 1. Analysis of the dataset 1 illustrating the consequences of genotyping errors and inconsistencies on QTL mapping. Nonpolymorphic markers and
markers with more than 50% missing genotypes were excluded to avoid excessive calculation time. a) Comparison of the known marker map (left) and
the genetic map estimated from observed RF (right), as calculated by est.map() and represented by plotMap() functions of R/qtl. Lines connect the
positions of each marker in the 2 maps. The estimated map is considerably expanded because of multiple genotype inconsistencies. b) Distribution of
the ratio between estimated and known distances between adjacent markers. Markers with known and calculated distances below 1 cM were removed
as they may lead to extremely small or large ratios. The expansion of the estimated map leads to a distribution tail of high ratios. The y-axis is in
logarithmic scale. Fifty-seven percent of markers have a ratio above 5 (dashed line). c) Output of the scanone function of R/qtl showing the
identification of narrow LOD score peaks. Genome-wide significance thresholds computed by data permutation are shown as plain (P ¼ 0.05), dotted
(P ¼ 0.1), and dashed (P ¼ 0.63) lines. d) Magnification of the scanone plot restricted to chromosome 13 (peak p2). The LOD score peak is located on one
marker (red tick) distant by 1.728 and 1.24 cM from the proximal and distal markers, respectively, on the known marker map, but by 1,001.582 and
1,001.506 cM based on calculated RF. Downloaded from https://academic.oup.com/g3journal/advance-article/doi/10.1093/g3journal/jkac219/6674511 by Institut (a) (b)
(d) (a) (b) Downloaded from https://academic.oup.com/g3journal/advance-article/doi/10.1093/g3journal/jkac219/6674511 by Institut Pasteur - CeRIS user on 28 September 2022 Downloaded from https://academic.oup.com/g3journal/advance-article/doi/10.1093/g3journal/jkac219/6674511 by Institut Pasteur - CeRIS user on 28 September 2022 (c) (c) (d) Fig. 1. Analysis of the dataset 1 illustrating the consequences of genotyping errors and inconsistencies on QTL mapping. Nonpolymorphic markers and
markers with more than 50% missing genotypes were excluded to avoid excessive calculation time. a) Comparison of the known marker map (left) and
the genetic map estimated from observed RF (right), as calculated by est.map() and represented by plotMap() functions of R/qtl. Lines connect the
positions of each marker in the 2 maps. Data importation Converting alleles into genotypes requires that SGI segregate
for the 2 parental alleles, and that each allele is found only in one
parent. The aim of the mark_allele() function is to control consis-
tency of allele’s origin at multiple levels. Genetic mapping requires both genotype and phenotype data. Required formats and instructions are detailed in the vignette
(see example of phenotype data in Supplementary Table 1). Parental strains’ genotyping data can be loaded from the same
genotyping results as the SGI, from a previous genotyping file or
from a reference file. Annotation data from K. Broman can be
imported directly from GitHub. The geno_strains() function for-
mats parental genotypes from a 2-allele encoding in Illumina for-
mat into a single letter encoding, and merges these data with the
annotation table into a table with parental allele and marker
positions. First, this function excludes markers with missing data in
both parents. If allele data are available for only one parent and
this allele is also found in SGI, the other allele present in SGI
will be assigned to the parent with missing allele. However, this
imputation is not error-free since we have observed, in rare
occasions, markers which alleles were identical in the parental
strains but were polymorphic in the SGI (Table 2 for such SNPs
in dataset 1). This situation may occur when the parental
strains used in the cross have diverged from those of the refer-
ence panel, or if one parent is heterozygous. Such markers will
be excluded by the mark_allele() function but they could escape
detection if allele information was missing in one parent. Consistency between parents and SGI alleles and genotypes
Several generations of MUGA arrays have been developed (Mega,
Giga, Mini), each with successive versions differing by multiple 4
|
G3, 2022, Vol. 00, No. 0 4
|
G3, 2022, Vol. 00, No. 0 Table 1. stuart analysis pipeline and application to dataset 1. Steps
Function
Excluded markers
Number of markers retained
1. Import SGI alleles from MUGA
arrays
read.table()/read_tsv()
–
11,125
2. Add data from parental strain
Genotyped with SGI: make consensus
geno_strains()
–
–
Imported from another dataset:
import and make consensus
read.table()/read_tsv(), geno_strains
–
–
Imported from reference
read.table()/other readr function
depending on the format
–
–
3. Filtering report and impact on QTL mapping results At every step, the markers filtered out are annotated in a marker
table which can be exported for further inspection. The last col-
umn of Table 1 shows the number of markers retained after each
step in the example dataset 1. Most of the starting markers
(7,180/11,125 ¼ 65%) which were eventually removed by stuart’s
functions were removed by mark_poly() as nonpolymorphic, a ra-
tio expected for crosses between 2 standard mouse inbred strains
(Frazer et al. 2007). mark_allele() rejected 750 markers, mark_na()
457 and mark_prop() 484. Across the 3 datasets, we found 1,546
markers with either non-Mendelian proportions or allele incon-
sistencies between parental strains and SGIs. Overall, 619 of
them were retained by stuart’s filtering in at least one of the Nonpolymorphic markers Genetic analysis requires polymorphic markers, i.e. for which
parents carry different alleles which segregate in the SGI. The
mark_poly() function excludes markers for which all genotyped
SGI carry the same allele, which saves computation time. Data importation Filter on allele consistency between
parents and SGI
Same set of markers between parents
and SGI
mark_match()
Not present in both parents and
SGI
11,125
Alleles consistent between parents
and SGI
mark_allele
Missing alleles in both parents
10,375
Not polymorphic in parents but
polymorphic in SGI
Different alleles in parents and
SGI
In backcrosses: homozygotes for
the wrong allele
Optional: one parent missing or
heterozygous
4. Exclude markers with high
proportion of missing genotypes
mark_na()
>50% of missing genotypes by
default
9,918
5. Exclude nonpolymorphic markers
in SGI
mark_poly()
Nonpolymorphic in SGI
2,738
6. Verify Mendelian proportions
mark_prop()
Departure from expected
Mendelian segregation (pro-
portion of each class or statis-
tical threshold)
2,254
7. Verify RF between markers
est.map() followed by mark_estmap()
High RFs with adjacent markers
2,251 failures may result from poor-quality genotyping assay. The mar-
k_na() function excludes such poorly genotyped markers. failures may result from poor-quality genotyping assay. The mar-
k_na() function excludes such poorly genotyped markers. Table 2. Markers of dataset 1 non polymorphic between parental
strains but polymorphic in SGI. Marker
Allele
parent 1
Allele
parent 2
Allele
SGI 1
Allele
SGI 2
S6J017555686
C
C
T
C
S6J113080150
G
G
A
G
gJAX00038569
C
C
T
C
mUNC21540855
C
C
A
C
gUNC21555204
T
T
T
C
gUNC21596600
A
A
A
G Table 2. Markers of dataset 1 non polymorphic between parental
strains but polymorphic in SGI. Mendelian proportions In 2-generation crosses between inbred strains, the proportions of
the 2 or 3 classes of genotypes are predictable, i.e. for autosomes,
25% of each type of homozygotes and 50% of heterozygotes in an
intercross, and 50% of homozygotes and 50% of heterozygotes in
a backcross. Comparing the observed proportions with these
expectations provides another criterion of filtering. The mark_prop() function filters markers based either on a
minimum proportion of each genotype or on the statistically sig-
nificant departure from the expected proportions (Chi2 test, with
a P-value threshold). Figure 2b shows the exclusions of the auto-
somal markers depending on the proportion of each genotype. X
chromosome
genotypic
proportions
differ
from
autosomes,
therefore, different arguments of mark_prop() function are used
to filter X-linked markers for more precise curation. Adding the parNH ¼ FALSE argument to the mark_allele() func-
tion will exclude markers missing one parental allele or for
which one parent is heterozygous. However, while preventing
rare errors, this option will also exclude a number of truly infor-
mative markers. The mark_allele() function also discards markers at which
parents and SGI carry different alleles, and, for backcrosses,
markers for which some SGI are homozygous for the wrong allele. Missing genotypes Reliable QTL mapping results depend on markers with medium
to high rate of successful genotyping. Figure 2a shows markers
distribution based on the proportion of missing genotypes. For
over 95% of markers genotyping rate was above 50%. Genotyping M. Bourdon and X. Montagutelli 5 1
10
100
1000
0.00
0.25
0.50
0.75
1.00
Proportion of NA
Number of markers
Proportion of missing genotypes
(a)
0.00
0.25
0.50
0.75
1.00
0.00
0.25
0.50
0.75
1.00
Proportion of A/A individuals
Proportion of B/B individuals
Retained
Excluded
Exclusion of markers by mark_prop()
(b)
Fig. 2. a) Distribution of the markers by their proportion of missing
genotype (NA) in dataset 1. The y-axis is in logarithmic scale. 4.63% of
markers have >50% missing genotypes. b) Exclusion of markers
depending on genotypic proportions in dataset 1. Markers on X and Y
chromosomes and mitochondrial DNA are not represented. The 2 axes
represent the proportions of the 2 types of homozygous individuals in
the intercross: AA and BB. Each dot represents a marker. Markers were
excluded if the proportion of at least one of the 3 genotypes (AA, AB, and
BB) was less than 10%, i.e. outside the triangle defined by the 3 dashed
lines (AA ¼ 0.1, BB ¼ 0.1, and AA þ BB ¼ 0.9). Arrows point at 2 markers
excluded due to a proportion of heterozygotes <10%. adjacent markers lower than 2 cM, which is largely sufficient to
perform QTL mapping (Darvasi et al. 1993). After curation, pheno-
type and genotype data are combined and exported in the R/qtl
format using the write_rqtl() function. The qtl2convert package
(Broman 2021) converts this output into the adequate format re-
quired by the more recent R/qtl2 package. 1
10
100
1000
0.00
0.25
0.50
0.75
1.00
Proportion of NA
Number of markers
Proportion of missing genotypes
(a) Proportion of missing genotypes (a) Downloaded from https://academic.oup.com/g3journal/advance-article/doi/10.1093/g3journal/jkac219/6674511 by Institut Pasteur - CeRIS user on 28 September 2022 Downloaded from https://academic.oup.com/g3journal/advance-article/doi/10.1093/g3journal/jkac219/6674511 by Institut Pasteur - CeRIS user on 28 September 2022 Figure 3a and Supplementary Fig. 4, a and b show the marker
maps calculated after data curation with stuart. The known
marker map and the estimated genetic map are consistent, with
minimal expansions or contractions. Large ratios between the
calculated and the known genetic distances between adjacent
markers have been eliminated (Fig. 3b, Supplementary Fig. 4, c
and d). Missing genotypes QTL mapping analysis on curated dataset 1 is shown on
Fig. 3c (to be compared with Fig. 1c; see Supplementary Fig. 5 for
datasets 2 and 3). LOD thresholds are in the expected range for
an F2, and the LOD score curve reveals broader peaks than in
Fig. 1b, with progressive LOD score decrease on both sides of the
peak marker. One significant and 3 suggestive QTLs were identi-
fied on chromosomes 12 (P-value ¼ 0.037, Fig. 3d), 5 (P-val-
ue ¼ 0.460), 10 (P-value ¼ 0.157), and 15 (P-value ¼ 0.244) which
were not visible using noncurated data due to very high LOD
score thresholds. 1
10
0.00
0.25
0.50
0.75
1.00
Proportion of NA
0.00
0.25
0.50
0.75
1.00
0.00
0.25
0.50
0.75
1.00
Proportion of A/A individuals
Proportion of B/B individuals
Retained
Excluded
Exclusion of markers by mark_prop()
(b)
Fig. 2. a) Distribution of the markers by their proportion of missing
genotype (NA) in dataset 1. The y-axis is in logarithmic scale. 4.63% of
markers have >50% missing genotypes. b) Exclusion of markers
depending on genotypic proportions in dataset 1. Markers on X and Y
chromosomes and mitochondrial DNA are not represented. The 2 axes
represent the proportions of the 2 types of homozygous individuals in
the intercross: AA and BB. Each dot represents a marker. Markers were
excluded if the proportion of at least one of the 3 genotypes (AA, AB, and
BB) was less than 10%, i.e. outside the triangle defined by the 3 dashed
lines (AA ¼ 0.1, BB ¼ 0.1, and AA þ BB ¼ 0.9). Arrows point at 2 markers
excluded due to a proportion of heterozygotes <10%. p
0.00
0.25
0.50
0.75
1.00
0.00
0.25
0.50
0.75
1.00
Proportion of A/A individuals
Proportion of B/B individuals
Retained
Excluded
Exclusion of markers by mark_prop()
(b) Exclusion of markers by mark_prop() 1
(b) Being very simple to use and efficient at curating genotyping
errors, stuart will facilitate the use of genotyping arrays for ge-
netic mapping purposes in 2-generation crosses, bridging the gap
between raw allele data produced by SNP platforms and genetic
analysis software. Moreover, its functions can be used indepen-
dently to analyze inbred strains genotypes. For example, geno_-
strain() creates a genotype consensus between 2 or more
individuals of the same strain suitable for further inspection,
which can be useful when genotyping or regenotyping a strain of
interest. Data availability All datasets used as examples in this article are available from
https://gitlab.pasteur.fr/mouselab/stuart/. Dataset 1 is included
in the package and can be loaded once the package is loaded (see
the vignette for details). The 2 other datasets are available from
GitLab in the “article” directory in separate folders (i.e. “data2”
and “data3”). Each folder contains the genotypes of the SGIs in
file
“geno_dataX.csv,”
the
phenotypes
of
the
SGIs
in
file
“pheno_dataX.csv,”
the
parental
strains’
genotypes
in
file
“parents_dataX.csv” and the reference genotypes for the parental
strains in file “ref_geno_dataX.csv.” Analysis of each cross is in
each folder in an R markdown file (“dataX.Rmd”). other crosses, ruling out their misassignment to the genetic map. Out of the residual markers, 85 were removed from all datasets
for another criterion than absence of polymorphism and were
therefore considered as unreliable. At this step, the dataset may still contain markers showing
high RFs with adjacent markers either for a reason not tested by
the current version of stuart or due to the parameters used in
mark_na() and mark_prop() functions. These markers can be
identified by calculating the estimated map using R/qtl est.map()
and using stuart’s mark_estmap() function which excludes
markers presenting high RFs with adjacent markers. Over the 3
datasets, 9 markers were removed by mark_estmap(). Five of
them were retained in at least one other dataset, indicating the
problem was dataset specific. Finally, for dataset 1, 2,251 markers
passed all steps resulting in an average genetic interval between Supplemental material is available at G3 online. Supplemental material is available at G3 online. Web resources The source code of the stuart package and the code used for the
figures of this article are publicly available from https://gitlab.pas
teur.fr/mouselab/stuart/. Missing genotypes Comparing genotyping results of an inbred strain after
several generations of breeding with mark_allele() will readily
identify variants that have emerged or been selected over time. Likewise, this function will help identifying genetic variants be-
tween substrains. Conflicts of interest None declared. None declared. Darvasi A, Weinreb A, Minke V, Weller JI, Soller M. Detecting marker-
QTL linkage and estimating QTL gene effect and map location us-
ing a saturated genetic map. Genetics. 1993;134(3):943–951. doi:
10.1093/genetics/134.3.943. Acknowledgments The marker with the highest
LOD score is identified with a thick (red) tick. Fig. 3. Analysis of dataset 1 after curation of genotyping data by stuart using the mark_match(), mark_allele(), mark_na(), mark_poly(), mark_prop(),
and mark_estmap() functions. Refer to Fig. 1 for comparison with original data. a) The estimated marker map is now consistent with the known marker
map. Despite some contraction or expansion of specific intervals, the genome length of the observed marker map for each chromosome is consistent
with the known map (ratio between the calculated and the known length of the genome: 1.12). b) The distribution of the ratio between estimated and
known distance between adjacent markers. Markers with known and calculated distances below 1 cM were removed as they may lead to extremely
small or large ratios. Ratios are normally distributed with mean ¼ 1.33 and SD ¼ 0.81 showing consistency between the known and estimated maps. c) The LOD score curve shows several peaks, one of which is significant at P < 0.05 (plain line, genome-wide significance computed by data
permutation). Note that the significance thresholds are much lower than in Fig. 1c. None of the peaks shown in Fig. 1c were confirmed after data
curation. Conversely, none of the peaks above P ¼ 0.63 (dashed line) found after data curation had been detected in Fig. 1c. d) The magnification of the
QTL peak identified on chromosome 12, showing progressive decrease of the LOD score curve over a large genetic interval. The marker with the highest
LOD score is identified with a thick (red) tick. Funding Broman KW, Gatti DM, Simecek P, Furlotte NA, Prins P, Sen S, Yandell
BS, Churchill GA. R/qtl2: software for mapping quantitative trait
loci with high-dimensional data and multiparent populations. Genetics. 2019;211(2):495–502. doi:10.1534/genetics.118.301595. This
project
was
funded
by
the
French
Government’s
Investissement d’Avenir programme, Laboratoire d’Excellence
“Integrative Biology of Emerging Infectious Diseases” (grant
nANR-10-LABX-62-IBEID). Broman KW, Wu H, Sen S, Churchill GA. R/qtl: QTL mapping in experi-
mental crosses. Bioinformatics. 2003;19(7):889–890. doi:10.1093/
bioinformatics/btg112. Cheung CYK, Thompson EA, Wijsman EM. Detection of Mendelian con-
sistent genotyping errors in pedigrees: detection of genotyping
errors. Genet Epidemiol. 2014;38(4):291–299. doi:10.1002/gepi.21806. Acknowledgments b) The distribution of the ratio between estimated and
known distance between adjacent markers. Markers with known and calculated distances below 1 cM were removed as they may lead to extremely
small or large ratios. Ratios are normally distributed with mean ¼ 1.33 and SD ¼ 0.81 showing consistency between the known and estimated maps. c) The LOD score curve shows several peaks, one of which is significant at P < 0.05 (plain line, genome-wide significance computed by data
permutation). Note that the significance thresholds are much lower than in Fig. 1c. None of the peaks shown in Fig. 1c were confirmed after data
curation. Conversely, none of the peaks above P ¼ 0.63 (dashed line) found after data curation had been detected in Fig. 1c. d) The magnification of the
QTL peak identified on chromosome 12, showing progressive decrease of the LOD score curve over a large genetic interval. The marker with the highest
LOD score is identified with a thick (red) tick. Fig. 3. Analysis of dataset 1 after curation of genotyping data by stuart using the mark_match(), mark_allele(), mark_na(), mark_poly(), mark_prop(),
and mark_estmap() functions. Refer to Fig. 1 for comparison with original data. a) The estimated marker map is now consistent with the known marker
map. Despite some contraction or expansion of specific intervals, the genome length of the observed marker map for each chromosome is consistent
with the known map (ratio between the calculated and the known length of the genome: 1.12). b) The distribution of the ratio between estimated and
known distance between adjacent markers. Markers with known and calculated distances below 1 cM were removed as they may lead to extremely
small or large ratios. Ratios are normally distributed with mean ¼ 1.33 and SD ¼ 0.81 showing consistency between the known and estimated maps. c) The LOD score curve shows several peaks, one of which is significant at P < 0.05 (plain line, genome-wide significance computed by data
permutation). Note that the significance thresholds are much lower than in Fig. 1c. None of the peaks shown in Fig. 1c were confirmed after data
curation. Conversely, none of the peaks above P ¼ 0.63 (dashed line) found after data curation had been detected in Fig. 1c. d) The magnification of the
QTL peak identified on chromosome 12, showing progressive decrease of the LOD score curve over a large genetic interval. Acknowledgments We thank Elise Jacquemet of the Pasteur Institute Bioinformatics
and Biostatistics HUB for helping with the use of GitLab. 6
|
G3, 2022, Vol. 00, No. 0 (a)
(b)
(c)
(d)
Fig. 3. Analysis of dataset 1 after curation of genotyping data by stuart using the mark_match(), mark_allele(), mark_na(), mark_poly(), mark_prop(),
and mark_estmap() functions. Refer to Fig. 1 for comparison with original data. a) The estimated marker map is now consistent with the known marker
map. Despite some contraction or expansion of specific intervals, the genome length of the observed marker map for each chromosome is consistent
with the known map (ratio between the calculated and the known length of the genome: 1.12). b) The distribution of the ratio between estimated and
known distance between adjacent markers. Markers with known and calculated distances below 1 cM were removed as they may lead to extremely
small or large ratios. Ratios are normally distributed with mean ¼ 1.33 and SD ¼ 0.81 showing consistency between the known and estimated maps. c) The LOD score curve shows several peaks, one of which is significant at P < 0.05 (plain line, genome-wide significance computed by data
permutation). Note that the significance thresholds are much lower than in Fig. 1c. None of the peaks shown in Fig. 1c were confirmed after data
curation. Conversely, none of the peaks above P ¼ 0.63 (dashed line) found after data curation had been detected in Fig. 1c. d) The magnification of the
QTL peak identified on chromosome 12, showing progressive decrease of the LOD score curve over a large genetic interval. The marker with the highest
LOD score is identified with a thick (red) tick. (b)
(d) (a) (a) (b) Downloaded from https://academic.oup.com/g3journal/advance-article/doi/10.1093/g3journal/jkac219/6674511 by Institut Pasteur - CeRIS user on 28 September 2022 Downloaded from https://academic.oup.com/g3journal/advance-article/doi/10.1093/g3journal/jkac219/6674511 by Institut Pasteur - CeRIS user on 28 Septemb (c)
(d (d) (c) (d) Fig. 3. Analysis of dataset 1 after curation of genotyping data by stuart using the mark_match(), mark_allele(), mark_na(), mark_poly(), mark_prop(),
and mark_estmap() functions. Refer to Fig. 1 for comparison with original data. a) The estimated marker map is now consistent with the known marker
map. Despite some contraction or expansion of specific intervals, the genome length of the observed marker map for each chromosome is consistent
with the known map (ratio between the calculated and the known length of the genome: 1.12). Broman KW. Review of statistical methods for QTL mapping in ex-
perimental crosses. Lab Anim (NY). 2001;30(7):44–52. Literature cited Broman KW. Review of statistical methods for QTL mapping in ex-
perimental crosses. Lab Anim (NY). 2001;30(7):44–52. Doerge RW, Churchill GA. Permutation tests for multiple loci affect-
ing a quantitative character. Genetics. 1996;142(1):285–294. doi:
10.1093/genetics/142.1.285. Doerge RW, Churchill GA. Permutation tests for multiple loci affect-
ing a quantitative character. Genetics. 1996;142(1):285–294. doi:
10.1093/genetics/142.1.285. Broman
KW. qtl2convert:
Convert
Data
among
QTL
Mapping
Packages. 2021. [accessed 2022 May 15]. https://CRAN.R-project. org/package=qtl2convert. Broman
KW. qtl2convert:
Convert
Data
among
QTL
Mapping
Packages. 2021. [accessed 2022 May 15]. https://CRAN.R-project. org/package=qtl2convert. Frazer KA, Eskin E, Kang HM, Bogue MA, Hinds DA, Beilharz EJ, Gupta
RV, Montgomery J, Morenzoni MM, Nilsen GB, et al. A sequence- Frazer KA, Eskin E, Kang HM, Bogue MA, Hinds DA, Beilharz EJ, Gupta
RV, Montgomery J, Morenzoni MM, Nilsen GB, et al. A sequence- Frazer KA, Eskin E, Kang HM, Bogue MA, Hinds DA, Beilharz EJ, Gupta
RV, Montgomery J, Morenzoni MM, Nilsen GB, et al. A sequence- M. Bourdon and X. Montagutelli Sigmon JS, Blanchard MW, Baric RS, Bell TA, Brennan J, Brockmann
GA, Burks AW, Calabrese JM, Caron KM, Cheney RE, et al. Content
and performance of the MiniMUGA genotyping array: a new tool
to improve rigor and reproducibility in mouse research. Genetics. 2020;216(4):905–930. doi:10.1534/genetics.120.303596. Gue´net JL, Benavides F, Panthier J-J, Montagutelli X. 2015. Genetics of
the Mouse. [accessed 2022 Jun 24]. https://link.springer.com/
book/10.1007/978-3-662–44287-6. Downloaded from https://academic.oup.com/g3journal/advance-article/doi/10.1093/g3journal/jkac219/6674511 by Institut Pasteur - CeRIS user on 28 September 2022 Svenson KL, Gatti DM, Valdar W, Welsh CE, Cheng R, Chesler EJ,
Palmer AA, McMillan L, Churchill GA. High-resolution genetic
mapping
using
the
mouse
diversity
outbred
population. Genetics. 2012;190(2):437–447. doi:10.1534/genetics.111.132597. Lander E, Kruglyak L. Genetic dissection of complex traits: guidelines
for interpreting and reporting linkage results. Nat Genet. 1995;
11(3):241–247. doi:10.1038/ng1195-241. Members of the Complex Trait Consortium. The nature and identifi-
cation of quantitative trait loci: a community’s view. Nat Rev
Genet. 2003;4(11):911–916. doi:10.1038/nrg1206. Wickham H. ggplot2: elegant Graphics for Data Analysis. 2016. [accessed 2022 May 15]. https://ggplot2.tidyverse.org. Morgan AP. argyle: an R package for analysis of illumina genotyping
arrays. G3 (Bethesda). 2015;6(2):281–286. doi:10.1534/g3.115.023739. Wickham H, Averick M, Bryan J, Chang W, McGowan L, Franc¸ois R,
Grolemund G, Hayes A, Henry L, Hester J, et al. Welcome to the tidy-
verse. J Open Source Softw. 2019;4(43):1686. doi:10.21105/joss.01686. Morgan AP, Fu C-P, Kao C-Y, Welsh CE, Didion JP, Yadgary L,
Hyacinth L, Ferris MT, Bell TA, Miller DR, et al. Communicating editor: F. P.-M. de Villena Literature cited The mouse univer-
sal
genotyping
array:
from
substrains
to
subspecies. G3
(Bethesda). 2015;6(2):263–279. doi:10.1534/g3.115.022087. Wickham H, Hester J, Chang W, Bryan J. devtools: Tools to Make
Developing R Packages Easier. 2021. [accessed 2022 May 15]. https://devtools.r-lib.org. Communicating editor: F. P.-M. de Villena
|
https://openalex.org/W4210322790
|
https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0263643&type=printable
|
English
| null |
Medical rehabilitation of older employees with migrant background in Germany: Does the utilization meet the needs?
|
PloS one
| 2,022
|
cc-by
| 9,341
|
PLOS ONE RESEARCH ARTICLE Chloe´ Charlotte Schro¨derID1*, Ju¨rgen Breckenkamp2, Jean-Baptist du Prel1 1 Department of Occupational Health Science, University of Wuppertal, Wuppertal, Germany, 2 Department
of Epidemiology & International Public Health, School of Public Health, Bielefeld University, Bielefeld,
Germany a1111111111
a1111111111
a1111111111
a1111111111
a1111111111 a1111111111
a1111111111
a1111111111
a1111111111
a1111111111 Abstract Due to demographic change with an ageing workforce, the proportion of employees with
poor health and a need for medical rehabilitation is increasing. The aim was to investigate if
older employees with migrant background have a different need for and utilization of medical
rehabilitation than employees without migrant background. To investigate this, self-reported
data from older German employees born in 1959 or 1965 of the first and second study wave
of the lidA cohort study were exploratory analyzed (n = 3897). Subgroups of employees with
migrant background were separated as first-generation, which had either German or foreign
nationality, and second-generation vs. the rest as non-migrants. All subgroups were exam-
ined for their need for and utilization of medical rehabilitation with descriptive and bivariate
statistics (chi-square, F- and post-hoc tests). Furthermore, multiple logistic regressions and
average marginal effects were calculated for each migrant group separately to assess the
effect of need for utilization of rehabilitation. According to our operationalizations, the foreign
and German first-generation migrants had the highest need for medical rehabilitation while
the German first- and second-generation migrants had the highest utilization in the bivariate
analysis. However, the multiple logistic model showed significant positive associations
between their needs and utilization of rehabilitation for all subgroups. Further in-depth analy-
sis of the need showed that something like under- and oversupply co-exist in migrant
groups, while the foreign first-generation migrants with lower need were the only ones with-
out rehabilitation usage. However, undersupply exists in all groups independent of migrant
status. Concluding, all subgroups showed suitable use of rehabilitation according to their
needs at first sight. Nevertheless, the utilization does not appear to have met all needs, and
therefore, the need-oriented utilization of rehabilitation should be increased among all
employees, e.g. by providing more information, removing barriers or identifying official need
with uniform standards. Medical rehabilitation of older employees
with migrant background in Germany: Does
the utilization meet the needs? hloe´ Charlotte Schro¨derID1*, Ju¨rgen Breckenkamp2, Jean-Baptist du Prel1 PLOS ONE PLOS ONE Introduction Due to the demographic change and prolonged working lives, the proportion of older employ-
ees is increasing in Germany [1, 2] and other European countries, and thus also the number of
employees with poor health and functional limitations [3]. Therefore, one major public health
goal in the next years and decades should be to avoid premature work exits due to poor health
with the help of primary prevention, rehabilitation and occupational re-integration. These will
gain relevance in working life, as e.g. medical rehabilitation is aiming at continuous active par-
ticipation in working life [4]. Additionally, medical rehabilitative services were implemented
within several guidelines in Germany over the years, e.g. for coronary heart disease [5]. Conse-
quently, there is a strong expectation that the needs and demands for rehabilitation will
increase in the future. Competing interests: The authors have declared
that no competing interests exist. Competing interests: The authors have declared
that no competing interests exist. In Germany, in order to be eligible for medical rehabilitation, the objective need must be
assessed first. The need for rehabilitation is not automatically officially acknowledged by the
psychological or physical impairment, but mainly from the continuing or expected
impairment of participation in social and working life [6–8]. The concerned person must submit an application himself, so that individual need can be
proven. The validation is jointly done by the rehabilitation providers (e.g. the pension, acci-
dent or the health insurance), who coordinate their responsibilities among themselves. Within
an objective socio-medical evaluation, information provided by the applicant, doctors, psycho-
logical psychotherapists and other therapeutic professions in social work and care are taken
into account. However, no uniform procedure to assess the objective need for rehabilitation
[9] exists and even the socio-medical evaluations seem to have only limited reliability [10]. Within rehabilitation research several more standardized assessment procedures were sug-
gested to support the identification of the need [7, 11], such as the “Luebecker algorithm” [8],
the “Work Ability Index” [12, 13], the “risk index for disability pension” [14, 15] or a “checklist
to identify the need for medical rehabilitation by general practitioners" [16, 17]. By now, the
latter is also recommended by the northern German pension insurance and provided to gen-
eral practitioners [17]. Within these assessments, working conditions and exposures and thus
the workability are linked to the need for rehabilitation. OPEN ACCESS Citation: Schro¨der CC, Breckenkamp J, du Prel J-B
(2022) Medical rehabilitation of older employees
with migrant background in Germany: Does the
utilization meet the needs? PLoS ONE 17(2):
e0263643. https://doi.org/10.1371/journal. pone.0263643 Editor: Marcel Pikhart, University of Hradec
Kralove: Univerzita Hradec Kralove, CZECH
REPUBLIC Copyright: © 2022 Schro¨der et al. This is an open
access article distributed under the terms of the
Creative Commons Attribution License, which
permits unrestricted use, distribution, and
reproduction in any medium, provided the original
author and source are credited. Data Availability Statement: The data underlying
the results presented in the study (lidA datasets of
the first and second wave) are available as a
Scientific Use File at the Research Data Centre of
the German Federal Employment Agency. Available
from: https://fdz.iab.de/en/FDZ_Individual_Data/
lidA.aspx. Data from the third wave will be added
by 2023. Funding: This research was funded by the
Deutsche Rentenversicherung Bund (grant 1 / 16 PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 PLOS ONE Need for and utilization of rehabilitation in older employees with migrant background in Germany number/funding code 0421 /40-64-50-61 lidA). The funders had no role in study design, data
collection and analysis, decision to publish, or
preparation of the manuscript. PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 Introduction Their proportion in the working population is continuously
increasing and has risen from 16.2% in 2010 to 24.4% in 2019. From these working EMB,
37.4% were > 45 years old in 2019, so part of the older working population [1, 2]. Based on these circumstances for foreigner, which are mostly G1 EMBs, one would assume
that they are likely to have a higher need for rehabilitation. So far, there are no studies investi-
gating this issue in EMBs or generally in the working population in Germany, as there is no
gold standard to assess the need for rehabilitation in Germany, yet. Those with foreign nationality, are more likely to retire earlier due to disability, compared
to employees with German nationality [23]. Such differences may be attributed to occupational
and health factors, but also to lower utilization of health services such as medical rehabilitation. Until 2018, studies showed that people with a migrant background are less likely to utilize
medical rehabilitation compared to those without (non-EMB) [27, 31–33], possibly due to bar-
riers such as lack of information, language problems, illiteracy, cultural aspects etc. [33–35]. However, there were no differences found in studies published in 2018 or later, so findings are
inconsistent and often lack information about the second-generation, because of the limited
differentiation of migrant background [33]. This lacking differentiation is a major limitation of other previous studies on migrants’
work, health or utilization of rehabilitation services in Germany. This is because quantitative
studies are often based on the analysis of secondary data such as process data. In such data
sets, it is mostly the feature “nationality” that allows for the differentiation of the migrant back-
ground. Yet, in Germany, such a definition leads to the misclassification of about half of all
people with a migrant background as non-migrants, as 11.1 million of a total 21.2 million peo-
ple with a migrant background, had German nationality in 2019 [2]. Additionally, primary
studies often do not make any further differentiation between migrant groups, even when
other operationalizations than nationality are used [33]. However, EMB are a heterogeneous
group and should be investigated in more detail. To our knowledge, even representative studies in Germany investigating the need for reha-
bilitation in older employees are missing and likewise for subgroups with migrant background. Introduction This is due to certain work exposures
increasing the risk of early retirement and disability pension which should be prevented with
the help of rehabilitation [18–21]. Therefore, the need for rehabilitation is related to the indi-
vidual workload. In particular, groups of employees who have worked as factory workers are burdened by
monotonous, repetitive work and physically demanding tasks [18, 19, 21]. In addition, psycho-
social workloads (e.g. low scope in decision-making, job insecurity, conflicts at work, time
pressure) are suspected to have an influence on the short and long-term probability of early
retirement due to illness [20]. Compared to those without a migrant background (non-EMB), employees with a migrant
background (EMB), especially foreign nationals, are more frequently exposed to such health-
endangering working conditions which our own data has also shown [22–25]. Compared to
non-EMB, EMB more often work as manual workers (semi-skilled and unskilled workers), i.e. they often work in low-skilled occupations and have less completed vocational training [22,
24]. Additionally, EMB are more frequently exposed to psychological workloads like lower
influence at work which all in all results in lower workability, significantly longer periods of
sick leave, more frequent occupational accidents and diseases (e.g. noise-induced hearing loss)
[22, 23, 25]. Additionally, when these unfavorable working conditions accumulate over work-
ing life, employees in higher working age might even be at higher risk for negative health out-
comes [18, 26]. However, the mentioned results mainly apply to foreigners or first-generation 2 / 16 PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 PLOS ONE Need for and utilization of rehabilitation in older employees with migrant background in Germany migrants, as we found in former analyses, second-generation migrants seem close to natives
[22, 25, 27]. Former research in this field likewise showed that descendants of immigrants have
fewer differences to the native population, probably due to adaption and different coping pro-
cesses while growing up in the host country. Reasons might be e.g. that they were not exposed
to the whole migration process themselves and have a higher utilization of social network as a
coping method compared to first-generation groups [22, 28–30]. The group of EMB comprises employees born outside of Germany (first-generation, G1)
and employees born in Germany, but with one or both parents born abroad (second-genera-
tion, G2) [2]. They can have German or foreign nationality, although the second-generation
mainly has German nationality. Introduction Furthermore, it is highly important to investigate the utilization of rehabilitation depending
on the need, to assess if the provision of health services like medical rehabilitation meets the
needs and demands in general. Therefore, the current study aimed to primarily investigate if subgroups of EMB have a dif-
ferent need for rehabilitation than non-EMB and secondly, if they use rehabilitation diver-
gently when considering their respective need for rehabilitation. Study design and participants The prospective lidA (leben in der Arbeit) cohort study investigates work, health and employ-
ment in older employees of two age cohorts (1959, 1965) as part of the “babyboomer 3 / 16 PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 PLOS ONE Need for and utilization of rehabilitation in older employees with migrant background in Germany generation” in Germany. This study is based on a representative two-stage random sample of
all socially insured employees of these cohorts in Germany in 2009. Due to the sampling speci-
fication, sworn civil servants and self-employed were not included. The participants were
interviewed at home for each assessment wave by computer assisted personal interviews
(CAPI), including a variety of questions about health, private life and work, as the participants
get closer to retirement. The baseline survey took place in 2011 (N = 6585), the second wave in
2014 (N = 4244) and the third wave in 2018 (N = 3586). All procedures performed in this study involving human participants were in accordance
with the ethical standards of the institutional and/or national research committee and with the
1964 Helsinki declaration and its later amendments or comparable ethical standards. Informed
verbal consent was obtained from all individual participants included in the study after
informing about study content, procedures and data protection in writing, according to good
epidemiological practice. This procedure has been approved by the Ethics Committee of the
University of Wuppertal (dated from 05/12/2008 and 20/11/2017, MS/BB 171025 Hasselhorn). The ethics approval refers to the whole lidA cohort study, not only this partial study. All the
lidA data was anonymized before starting analyses. A more detailed description of the lidA
cohort study including power calculation etc. can be found elsewhere [36]. Results of attrition analysis showed an almost selection-free realization of the sample in
relation to the sociodemographic characteristics used in the cited analyses [37–39] for all
waves. However, a more differentiated analysis revealed attrition of 65% for low educational
level in foreign and 63% in German G1 EMB compared to about 42% in non-EMB and 43% in
G2 EMB. Since this analysis included data from the first and second study wave, we performed
inverse probability weighting for subgroups of migrant status and educational level. The sam-
ple was restricted to those employed at least 1h/week in both study waves (N = 3961,
unweighted). Study design and participants Due to the weighting, cases with missing values in migrant background or edu-
cational level were also excluded Consequently the final sample consisted of 3897 individuals unweighted). Due to the weighting, cases with missing values in migrant background or edu-
cational level were also excluded. Consequently, the final sample consisted of 3897 individuals. PLOS ONE Table 1. Variables used in the lidA study, categorized according to the checklist of Deck et al. [16]. Original category of the checklist
Assigned variables of the lidA-study (self-reported)
Indication of rehabilitation: disease requiring treatment, chronification of
disease, comorbidities
• Incidence of disease requiring treatment (in the last 12 months)
• declared handicap/disability
Functional limitations: impairments in daily or working life
• poor physical health (lowest tertile of the SF-12 physical health scale, version of
the socio economic panel survey) [40, 41]
• frequent limitation due to pain (in the last 4 weeks) in daily life or at work
Accompanying psychological symptoms: depressiveness, anxiety, exhaustion
• poor mental health (lowest tertile of the SF-12 mental health scale, version of the
socio economic panel survey) [40, 41]
Influenceable risk factors: nicotine abuse, alcohol, lack of exercise, obesity,
dyslipidemia
• BMI > 30, BMI = weight/(height2)
• less/no sports or exercise in leisure time
• regular smoking at time of survey
Therapy: outpatient therapy not sufficient or not available nearby, intensification
required, unfavorable working hours
• working hours that are unfavorable for therapy (such as shift work, especially
night and alternating shifts)
Adverse influences in work, profession and everyday life: significant physical or
environmental work exposure e.g. heavy lifting, noise etc., psychological stress
• lower workability in relation to physical and mental job demands (second
dimension of the workability index, >8 points: normal work ability, <8 points: low
work ability) [42]
• high work stress (highest tertile of the effort-reward-imbalance ratio, indicating
high efforts but low rewards) [43, 44]
• more than one physical work exposure (e.g. heavy lifting and carrying; for at least
half of the working time)
Disability: current or threatened incapacity for work, long or repeated sick leave
in the last 2 years
• official sick leave > 30 days (in the last 12 months)
• officially declared reduced capacity to work or job-related incapacity
• indication of "prolonged illness" in the question about employment
Motivation and disease management: motivation to participate and to change
own lifestyle is present, own disease management strategies are insufficient
no variables from the first or second wave of the lidA-study can be assigned
https://doi.org/10.1371/journal.pone.0263643.t001 Table 1. Variables used in the lidA study, categorized according to the checklist of Deck et al. [16]. Operationalization Dependent variable. The outcome of the study was the self-reported “utilization of medi-
cal rehabilitation” indicated in the second study wave. Participants were asked to report
whether they had utilized an in- or outpatient rehabilitation service in the previous three years. The answers for in- and outpatient services were summarized vs. no utilization of rehabilita-
tion, generating a binary variable. Independent variable. The counterpart and other aspect of rehabilitation was the “need
for medical rehabilitation”, which we operationalized with the help of a summarizing score
taking different relevant aspects of life into account. A range of such variables was considered
in a checklist in a study by Deck et al. and is now recommended by the northern German pen-
sion insurance for general practitioners to assess the need for rehabilitation [16, 17]. This
checklist provided the basis for the summarizing score, so that representative and appropriate
variables of the lidA study were assigned to each category of the checklist (see Table 1). All
those self-reported variables were taken from the first study wave to consider need for and uti-
lization of rehabilitation consequentially over the course of time. If any of the mentioned vari-
ables applied to a person, then the item got the coding 1. At the end, there was a possible range
of values from 0 to 15, while summing up at least 10 valid items and allowing 5 missing items. The score correlated significantly with general health, the single item Short Form-12 Health
Survey (SF-12) [40], by rpbis = .568. Migrant background. The lidA cohort study allows to distinguish between migrant
groups by means of different specific indicators as proposed by Schenk et al. [45]. EMB were PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 4 / 16 Need for and utilization of rehabilitation in older employees with migrant background in Germany PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 Statistical analysis For interpretational purposes additional exploratory analysis were done to examine the uti-
lization depending on the need in more detail for each subgroup separately on bivariate level. For interpretational purposes additional exploratory analysis were done to examine the uti-
lization depending on the need in more detail for each subgroup separately on bivariate level. For this, the need score was divided into tertiles, in order to see the percentage of utilization
in people with lower, medium or higher need. For this, the need score was divided into tertiles, in order to see the percentage of utilization
in people with lower, medium or higher need. PLOS ONE defined based on the participants’ self-reported country of birth and nationality and on the
country of birth of each of their parents. Participants born in Germany, with German national-
ity and with both parents being born in Germany constitute the reference group (non-EMB). The group of EMB was divided in three subgroups to investigate potential differences. Firstly,
they were separated in generations, based on a definition provided by the German Federal Sta-
tistical Office [1, 2], so into first-generation (G1 EMB) and second-generation (G2 EMB), as
described before. Secondly, G1 EMB were divided into those with German and foreign nation-
ality, as in own (unpublished) pre-analyses, differences between these groups were detected. In
G2 EMB nearly all participants had German nationality, so these weren’t differentiated any
further. In the end, there were four groups: non-EMB vs. German G1 EMB, foreign G1 EMB
and G2 EMB. Covariates. To control for sociodemographic differences, the following variables were
considered as potential confounders when comparing groups with different migration back-
ground regarding the association of their need for and utilization of rehabilitation: Year of
birth (1959/1965), sex (male/female), and education. Education was operationalized with a
score combining school and professional education according to the recommendations of the
German Society of Epidemiology for the measurement and quantification of sociodemo-
graphic characteristics in epidemiological studies [46]. Accordingly, values from 1 (= not any
graduation) to 8 (= school leaving examination and graduation from college) were calculated
for each combination of school and professional education. For ease of interpretation the
score was classified in three categories: high, medium and low level education. 5 / 16 PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 PLOS ONE Need for and utilization of rehabilitation in older employees with migrant background in Germany Statistical analysis Due to group differences in attrition between the first and the second study wave relating to
migrant status and educational level, basic inverse probability weighting was used to account
for potential non-response bias. Inverse probability weighting is a method, where the data is
standardized on a certain population, which is different from the one, in which the data was
collected [47]. In our case the data was standardized on the population of the lidA baseline
assessment in 2011. For each subgroup the equation was: weight = percentage in wave 1/per-
centage in wave 2, so e.g. for the group of non-EMB with low education: 19.71%/17.89% =
1.1017. Simultaneously, the weighting factors were calculated for all other subgroups, which
can be found in S1 Table. All reported results are based on weighted analyses; however, in the
S2 Table unweighted characteristics are additionally presented for comparison. Descriptive and bivariate statistics including chi-square tests, F-tests within analyses of vari-
ance (ANOVA) and a Tukey post-hoc test were used to characterize the full sample and specif-
ically investigate differences between groups. For the multiple logistic regression analyses,
possible multicollinearities were determined as a pre-check using linear regression models of
the independent variable, utilization of rehabilitation. The results of the linear regression anal-
yses are not shown because no statistical evidence of multicollinearity was found. The inflation
of variance for all variables was 1.09. Tests for possible interactions of the need with the
sociodemographic covariates were done, which were all not statistically significant. Finally,
multiple logistic regressions were performed to investigate the influence of the need for the uti-
lization of medical rehabilitation for each migrant group separately. To further control for
sociodemographic differences, the logistic regressions were adjusted for sex, year of birth and
education in the full model. In all statistical tests p-values (two-tailed) <.05 were considered to be statistically signifi-
cant. These statistical analyses were performed using SPSS version 25.0 (IBM Corp.). In addition, average marginal effects (AMEs) were computed for all logistic regressions
with SAS 9.4. They allow to compare the results of nested models that otherwise may be biased
by unobserved heterogeneity. The AME shows for each variable in a regression model how
much the event probability changes when the independent variable increases by one unit, or
rather when a binary independent variable changes its level [48]. All multiple analyses were
done as complete case analyses. PLOS ONE Table 2. Characterization of study population (weighted samplea, n = 3897). Non-EMB (n = 3211)
German G1 EMB (n = 276)
Foreign G1 EMB (n = 130)
G2 EMB (n = 280)
p-valueb
Sex [n (%)]
Male
1481 (46.1)
129 (46.7)
61 (46.9)
121 (43.2)
.800
Female
1729 (53.9)
147 (53.3)
69 (53.1)
159 (56.8)
Year of birth [n (%)]
1959
1458 (45.4)
135 (48.9)
50 (38.5)
117 (41.8)
.152
1965
1753 (54.6)
141 (51.1)
80 (61.5)
163 (58.2)
Education level [n (%)]
High
663 (20.6)
55 (19.9)
33 (25.4)
61 (21.8)
< .001
Medium
1787 (55.7)
137 (49.6)
38 (29.2)
141 (50.4)
Low
761 (23.7)
84 (30.4)
59 (45.4)
78 (27.9)
Utilization of rehabilitation [n (%)], m = 3
Yes
390 (12.2)
48 (17.4)
14 (10.8)
48 (17.1)
.009
No
2818 (87.8)
228 (82.6)
116 (89.2)
233 (82.9)
EMB, employees with migrant background; G1, first-generation; G2, second-generation; m, number of missing values due to respondents not responding to the item,
from weighted results. a Total case numbers of each variable vary slightly because of rounding after weighting. b tested with Chi2-test. https://doi.org/10.1371/journal.pone.0263643.t002 Table 2. Characterization of study population (weighted samplea, n = 3897). EMB, employees with migrant background; G1, first-generation; G2, second-generation; m, number of missing values due to respondents not responding to the item,
from weighted results. However, they also had the highest proportion of high educational level with 25.4% compared
to the rest with around 20%. However, they also had the highest proportion of high educational level with 25.4% compared
to the rest with around 20%. Concerning the outcome of utilized rehabilitation, significant differences were likewise
observed (p = .009). The highest utilization was reported by German G1 EMB and G2 EMB
(around 17% respectively) and the lowest by foreign G1 EMB (10.8%) and non-EMB (12.2%). In contrast, foreign G1 EMB showed the highest need for rehabilitation when comparing
means of the need score (Fig 1). The mean values differed significantly between the four
groups as determined by one-way ANOVA [F(3, 3978) = 5.91, p <. 001, η2 = 0.004]. A post- Concerning the outcome of utilized rehabilitation, significant differences were likewise
observed (p = .009). The highest utilization was reported by German G1 EMB and G2 EMB
(around 17% respectively) and the lowest by foreign G1 EMB (10.8%) and non-EMB (12.2%). PLOS ONE In contrast, foreign G1 EMB showed the highest need for rehabilitation when comparing
means of the need score (Fig 1). The mean values differed significantly between the four
groups as determined by one-way ANOVA [F(3, 3978) = 5.91, p <. 001, η2 = 0.004]. A post-
hoc test revealed that the need for rehabilitation was statistically higher for German G1 EMB
(4.14 ± 2.48, p = 0.006) and foreign G1 EMB (4.23 ± 2.63, p = 0.043) compared to non-EMB
(3.66 ± 2.25). There was no statistically significant difference between G2 EMB and non-EMB
(p = 1.0). Descriptive and bivariate analysis In Table 2 the characteristics of all participants included in the analyses are presented, shown
as weighted results (n = 3897). Most of the participants were non-EMB (82.4%), around 7%
each German G1 EMB and G2 EMB, and the smallest group was foreign G1 EMB with 3.3%. Due to deliberate oversampling, participants born in 1965 were overrepresented in all sub-
groups. The same applied to female sex in all groups, the proportion of women was always
higher than for men. The distribution of educational level differed significantly between the
groups (p<.001), nearly half of foreign G1 EMB had low educational level (45.4%) while the
other groups had percentages between 23.7% (non-EMB) and 30.4% (German G1 EMB). PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 6 / 16 Need for and utilization of rehabilitation in older employees with migrant background in Germany G1, first-generation; G2, second-generation; m, number of missing values due to respondents not responding to the item, y slightly because of rounding after weighting. Multiple logistic regressions To answer the second research question, logistic regressions were conducted separately for
each migrant group to investigate further behavioral or migrant-group-specific differences
(see Table 3). In bivariate analyses, foreign G1 EMB showed the highest need for rehabilitation
(see Fig 1), but the lowest utilization of rehabilitation (see Table 2). To examine the association
for each group, the odds and the probability for using rehabilitation were calculated depending
on the need score. In all models, the need was positively associated with the utilization of reha-
bilitation in each group: the higher the need, the more likely the utilization. In the model adjusted for sex, year of birth and education, foreign G1 EMB had the highest
odds (OR 2.02, 95% CI 1.40–2.91) and the highest probability (4.2% for each unit change) to
utilize medical rehabilitation when the need increased. The other groups showed ORs from
1.25 to 1.30 each. When testing narrowed models, in detail only one sociodemographic control
variable at a time to follow the “one in ten rule”, nearly no change in the coefficients was
detected. 7 / 16 PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 PLOS ONE Need for and utilization of rehabilitation in older employees with migrant background in Germany Fig 1. Arithmetic mean values and 95%-confidence intervals of the need score for rehabilitation in migrant groups (weighted results, n = 3897). https://doi.org/10.1371/journal.pone.0263643.g001 Fig 1. Arithmetic mean values and 95%-confidence intervals of the need score for rehabilitation in migrant groups (weighted results, n = 3897). https://doi.org/10.1371/journal.pone.0263643.g001 Fig 1. Arithmetic mean values and 95%-confidence intervals of the need score for rehabilitation in migrant groups (weighted results, n = 3897). https://doi.org/10.1371/journal.pone.0263643.g001 Additional exploratory analysis for interpretation Our bivariate analyses revealed that foreign G1 EMB have a higher need but lower utilization
(Tables 1 and 2), which was presumed before analysis. To be able to interpret this apparent
contradiction, we examined the utilization depending on the need in more detail for each sub-
group separately on bivariate level (Table 4). Hereby, undersupply for all subgroups indepen-
dent from migrant background was detected to the extent that over 70% of the people with
higher need are not utilizing rehabilitation services. However, in three groups there are still
7–10% of those with lower need that have used rehabilitation in the past. Only in foreign G1
EMB those with lower need have not used rehabilitation at all. Discussion In the present study, we analyzed the need for and respective utilization of rehabilitation for
employee groups with and without migrant background. For the primary research question,
we identified that foreign and German G1 EMB had the highest need for rehabilitation when
measuring with our need score. The highest utilization of rehabilitation was reported by Ger-
man G1 EMB and G2 EMB with 17%, while foreign G1 EMB showed the lowest with 11%. Sec-
ondarily, when considering the respective need in multiple logistic regressions, significant 8 / 16 PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 PLOS ONE Need for and utilization of rehabilitation in older employees with migrant background in Germany Table 3. Stratified logistic regressions for the utilization of rehabilitation services depending on the need for reha-
bilitation and further sociodemographic variables (weighted results). Crude model: need
Full model: need + sex, year of birth, education
Non-EMB (n = 3208/ nevents = 390)
OR (95% CI)
1.24 (1.19–1.30)
1.25 (1.19–1.31)
AME
+0.0228
+0.0234
R2
0.052
0.054
German G1 EMB (n = 276/ nevents = 48)
OR (95% CI)
1.22 (1.08–1.38)
1.25 (1.10–1.43)
AME
+0.0273
+0.0289
R2
0.058
0.084
Foreign G1 EMB (n = 127/ nevents = 14)
OR (95% CI)
1.65 (1.27–2.13)
2.02 (1.40–2.91)
AME
+0.0318
+0.0421
R2
0.276
0.353
G2 EMB (n = 279/ nevents = 48)
OR (95% CI)
1.27 (1.11–1.46)
1.30 (1.12–1.50)
AME
+0.0310
+0.0327
R2
0.070
0.087
p < .05,
p < .01,
p < .001. AME, average marginal effects; CI, confidence interval; M, Model; nevents, number of events where the outcome = 1
in the logistic regression; OR, Odds Ratio; p, p-value; Ref., Reference; R2, Nagelkerke pseudo-R2. https://doi.org/10.1371/journal.pone.0263643.t003 p
AME, average marginal effects; CI, confidence interval; M, Model; nevents, number of events where the outcome = 1
in the logistic regression; OR, Odds Ratio; p, p-value; Ref., Reference; R2, Nagelkerke pseudo-R2. p
AME, average marginal effects; CI, confidence interval; M, Model; nevents, number of events where the outcome = 1
in the logistic regression; OR, Odds Ratio; p, p-value; Ref., Reference; R2, Nagelkerke pseudo-R2. https://doi.org/10.1371/journal.pone.0263643.t003 positive associations with the utilization were found for each subgroup. Foreign G1 EMB
showed the highest association between need and utilization. However, the results of foreign G1 EMB have to be carefully interpreted as the case number
of this group was quite low (Table 3), as well as the number of “utilized rehabilitation” within
multiple logistic regression. PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 PLOS ONE Table 4. Utilization of rehabilitation services depending on the need for rehabilitation (separated for each group, weighted results, row percent, n = 3894). Need (tertiles)
Utilization
p-valuea
No
Yes
Non-EMB(n = 3208)
Lower
92.4%
7.6%
< .001
Medium
90.4%
9.6%
Higher
80.5%
19.5%
German G1 EMB (n = 276)
Lower
89.2%
10.8%
.001
Medium
89.0%
11.0%
Higher
71.6%
28.4%
Foreign G1 EMB (n = 130)
Lower
100%
0%
.011
Medium
90.5%
9.5%
Higher
80.0%
20.0%
G2 EMB (n = 280)
Lower
92.8%
7.2%
.009
Medium
82.7%
17.3%
Higher
75.8%
24.2%
EMB, employees with migrant background; G1, first-generation; G2, second-generation. a tested with Chi2-test. EMB, employees with migrant background; G1, first-generation; G2, second-generation. a tested with Chi2-test. EMB, employees with migrant background; G1, first-generation; G2, second-generation. a tested with Chi2-test. low need, compared to the other groups in Table 4. They might only utilize rehabilitation
when really necessary and their health situation is worsening. In more detail, our additional analysis for interpretational purposes detected that over 70%
of the people with higher need did not utilize rehabilitation services but instead 7–10% of
those with lower need used rehabilitation in the past. Only those with lower need did not use
rehabilitation at all in foreign G1 EMB. This raises the question as to whether health services
like medical rehabilitation are truly authorized according to the need in Germany. Especially
in older working age, health services like medical rehabilitation should be provided depending
on the existing need. Only in this way, equal opportunities to stay healthy and actively in work
and prevent early exit can be assured and unnecessary costs avoided. While there have been
some projects in Germany in the past, aiming at improving the information about and access
to medical rehabilitation where needed [e.g. 49], according to our result further efforts would
be worthwhile. The accessibility to medical rehabilitation in general might be improved by fur-
ther information campaigns or reducing formal access barriers (e.g. application process, wait-
ing times, travel distances or charges for those with lower/no income) with diversity in mind. More migrant-specific strategies to reduce language or cultural barriers would be important as
well. However, the main dependent factor is, of course, the instrument to assess the need for
rehabilitation, as the term “need” is not distinct and results are highly dependent of the chosen
instrument. Discussion This is due to loss-to-follow up between the first and second study
wave, which was weighted for, but also due to lower participation rate among foreigners in
general in the first study wave. If the case numbers had been higher, also for German G1 EMB
and G2 EMB, differences between confidence intervals (CI) of the four groups would probably
have been more precise, as the CI would get narrower. However, based on the confidence
interval of the OR in the adjusted model, our study only showed significant differences
between foreign G1 EMB and non-EMB while foreign G1 EMB have higher probability to uti-
lize medical rehabilitation than non-EMB when the need increased. This analysis is the first in Germany to identify the need for rehabilitation in different
migrant groups compared to non-migrants. The findings of a higher need for rehabilitation in
G1 EMB and especially in foreign G1 EMB match our assumptions before analysis as this
group often experience unfavorable working conditions, as mentioned in the introduction. Our results showed, when assessing need with the help of our need score: the higher the need,
the higher the utilization of rehabilitation for all groups. However, further barriers for instant
utilization of healthcare and rehabilitation e.g. language problems, illiteracy or cultural aspects
might exist for foreign G1 EMB due to own migration experiences and potential different
health beliefs within their cultural background. Hence, when assessing need for medical reha-
bilitation with our need score, this group showed zero utilization of rehabilitation when having PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 9 / 16 Need for and utilization of rehabilitation in older employees with migrant background in Germany PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 https://doi.org/10.1371/journal.pone.0263643.t004 PLOS ONE As described before, there are several operationalizations within rehabilitation
research in Germany suggested to support the identification of need. In the presented study,
we decided to orientate the operationalization towards the checklist of Deck et al. [16], as it
covers various life aspects of the person affected: Incidence of disease, functional limitations,
psychological factors, other risk factors such as smoking, motivation and coping with the dis-
ease, therapy, inability to work and impairments in work and everyday life. It convinced us
that the checklist is nowadays recommended by the northern German pension insurance for
general practitioners to assess the need for rehabilitation [17]. In our case however, the items
of the need score were based on subjective information of the study participants and not on an
objective assessment of the need for rehabilitation. Nonetheless, the items were collected PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 10 / 16 PLOS ONE Need for and utilization of rehabilitation in older employees with migrant background in Germany independently and without the purpose of assessing the need for rehabilitation. Such a check-
list or scoring not only helps general practitioners to screen their patients, but could addition-
ally help the official need assessment within the socio-medical evaluation, In the future, the operationalization should definitely be standardized and so we are calling
for a harmonization of the assessment procedure for the need of rehabilitation as other rehabil-
itation researchers [7, 8]. As rehabilitation is oriented towards the biopsychosocial model of ill-
ness and health, which is the basis of the International Classification of Functioning, Disability
and Health (ICF) [50, 51], this could be another approach for a standardized need assessment. It is already stated in the social code IX in Germany that the determination of the need for
rehabilitation should be carried out by an instrument that is based on the ICF, while also con-
sidering different life aspects e.g. mobility, domestic life, communication etc. [52]. Besides,
research has already found out that the ICF Generic 6 score is a valid tool to assess functioning
in several clinical settings [53], so this could be another possible instrument to use for need
assessment. To our knowledge, further research is still going on to implement the ICF in other
settings and test its practicability and reliability there [e.g. 54–57]. PLOS ONE Another important aspect, especially for the further outlook, constitutes the timing of needs
assessment, as people with need should be identified and allocated early enough to medical
rehabilitation. Schlo¨ffel and colleagues [58] tested an intervention of a web-based self-test to
identify need for rehabilitation and subsequently the effectiveness on the application rate. The
self-test was based on WAI and IMET (“Index to measure restrictions of participation”). Though, this intervention showed no significant effect as the only means, as Spanier and
Bethge also investigated [49, 59]. A solution could be to combine different means. Bethge and
his team already proposed in 2012 [12] a 3-staged procedure with screening of register data
using a validated risk index at first [15], then postal screening with WAI for persons with high
risk in the first step and lastly giving them consultation and information for the application for
rehabilitation. This procedure would be more likely to improve application rates [58] and the
utilization of rehabilitation according to personal need in the long run. However, this proce-
dure is not implemented yet within the German pension insurance or other rehabilitation pro-
viders, as far as we are informed. PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 Strengths & limitations Our study has several strengths. First, the sample is representative for the German population
of socially insured employees of the considered two age cohorts [37, 38]. Second, the lidA
study has the strength to differentiate more detailed subgroups with migrant background. Dif-
ferent indicators to map migrant status are used as recommended by Schenk et al. [45], not
only nationality. Another strength is the variety of the study characteristics, so that several
important factors to measure the need of rehabilitation could be taken into account. Here, vali-
dated instruments like WAI or SF12 were used to represent the different areas of life which are
considered in the checklist of Deck et al. [16]. In earlier rehabilitation research these were
already associated with the need for rehabilitation, however it remains an open question
whether the summing score to assess the objective need for rehabilitation is the right instru-
ment, as there is no gold standard in Germany so far. Another advantage was the ability to
consider the need for and utilization of rehabilitation in logical time order, as the need was
assessed from the first study wave and the utilization in the second study wave. Of course, no
causality can be proven in a study like this. Despite these strengths, the study also has its limitations. First, there is no gold standard for
the assessment of the need for medical rehabilitation in Germany. So, we could not test the
validity of our assessment instrument derived from the check list suggested by Deck et al. PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 11 / 16 PLOS ONE Need for and utilization of rehabilitation in older employees with migrant background in Germany (2009) for general practitioners more comprehensively. Furthermore, certain aspects which
influence the official need were discussed in previous rehabilitation research, but could not be
considered, as there were no suitable items within the first and second lidA study wave
assessed. These include the overall rehabilitation prognosis, the participant’s motivation and
therapy options, such as the local infrastructure for rehabilitation. Yet, these factors might be
more relevant in estimating the long-term success of medical rehabilitation and to a lesser
degree in assessing the actual need for rehabilitation, which was the focus of our investigation. Overall, the items of the need score were based on subjective information of the study partici-
pants and not on an objective assessment of the need for rehabilitation. Supporting information S1 Table. Weighting factors for inverse probability weighting. (DOCX) S2 Table. Characterization of study population (unweighted sample, n = 3944). (DOCX) Conclusions According to our results and operationalizations, all subgroups showed suitable use of rehabili-
tation according to their needs at first sight, foreign G1 showed the highest association. How-
ever, when looking more in detail, something like under- and oversupply co-exist in all
subgroups, while foreign G1 employees with lower need were the only ones without rehabilita-
tion usage. Yet, undersupply exists in all groups independent of migrant status. Therefore, the
need-oriented utilization of rehabilitation should be increased among all employees, e.g. by
providing more information, removing barriers or identifying official need with the same stan-
dards. The findings highlight the necessity for distinct operationalization and investigation of
migrant groups in research and resulting policy. Strengths & limitations Nonetheless, the items
were collected independently and without this purpose. Another limitation is the restriction to
two age cohorts within socially insured employees due to sampling, where the lidA study does
not include sworn civil servants and self-employed persons. While the majority of employees
in Germany are socially insured [2, 36], employees with migrant background are underrepre-
sented in the group of civil servants and overrepresented in the group of self-employed. So, the
health status of migrants and non-migrants could be different if civil servants and self-
employed were included. Consequently, the findings of this study are limited to socially
insured employees born in 1959 or 1965. Yet, overall there are at least comparable percentages
of different migrant backgrounds in the lidA-study in comparison to the German microcensus
[2]. Further restriction to generalizability could have been introduced by language bias
through the conduction of the study in German, where EMB were potentially excluded when
having language problems. Finally, as mentioned before, the case number of foreign G1 EMB
was quite low so that the results for this group have to be interpreted carefully. References 1. Statistisches Bundesamt, editor. Bevo¨lkerung mit Migrationshintergrund. Ergebnisse des Mikrozensus
2010—hochgerechnet auf Basis des Zensus 2011. Sonderausgabe der Fachserie 1 Reihe 2.2. Berlin:
Statistisches Bundesamt; 2017 [cited 2021 Jan 22]. https://www.destatis.de/DE/Themen/Gesellschaft-
Umwelt/Bevoelkerung/Migration-Integration/Publikationen/Downloads-Migration/
migrationshintergrund-sonderausgabe-5122121109004.pdf?__blob=publicationFile 2. Statistisches Bundesamt, editor. Bevo¨lkerung und Erwerbsta¨tigkeit. Bevo¨lkerung mit Migrationshinter-
grund—Ergebnisse des Mikrozensus 2019 -. Fachserie 1 Reihe 2.2. Berlin: Statistisches Bundesamt;
2020 [cited 2021 Jan 22]. https://www.destatis.de/DE/Themen/Gesellschaft-Umwelt/Bevoelkerung/
Migration-Integration/Publikationen/Downloads-Migration/migrationshintergrund-2010220197004.pdf? __blob=publicationFile 3. van den Berg T, Schuring M, Avendano M, Mackenbach J, Burdorf A. The impact of ill health on exit
from paid employment in Europe among older workers. Occupational and Environmental Medicine. 2010; 67:845–52. https://doi.org/10.1136/oem.2009.051730 PMID: 20798020 4. Buschmann-Steinhage R, Widera T. Grundlagen der Rehabilitation. In: Bengel J, Mittag O, editors. Psy-
chologie in der medizinischen Rehabilitation: Ein Lehr- und Praxishandbuch. Berlin, Heidelberg:
Springer Berlin Heidelberg; 2016. pp. 13–24. 5. A¨ rztliches Zentrum fu¨r Qualita¨t in der Medizin, editor. NVL Chronische KHK: Was ist wichtig? Was ist
neu? [Internet]. Berlin. 2019 [cited 2020 Nov 13]. https://www.leitlinien.de/mdb/downloads/nvl/khk/khk-
5aufl-flyer.pdf 6. Viehmeier S, Schubert M, Thimmel R. Vor der Rehabilitation. In: Bundesarbeitsgemeinschaft fu¨r Reha-
bilitation e. V., editor. Rehabilitation—Vom Antrag bis zur Nachsorge: fu¨r A¨ rzte, Psychologische Psy-
chotherapeuten und andere Gesundheitsberufe. Berlin: Springer; 2018. pp. 181–95. 7. Morfeld M. Identifikation von Reha-Bedarf. In: Weber A, Peschkes L, Boer Wd, editors. Return to work
—Arbeit fu¨r alle. Grundlagen der beruflichen Reintegration. 1st ed. Stuttgart: Genter Verlag; 2015. pp. 179–85. 8. Raspe H, Ekkernkamp M, Matthis C, Raspe A, Mittag O. Bedarf an rehabilitativen Leistungen. Theorie
und Empirie. Rehabilitation (Stuttg). 2005; 44:325–34. https://doi.org/10.1055/s-2005-915309 PMID:
16320176. 9. Thielgen G, Seel H. Strukturelle Grundlagen der Rehabilitation. In: Bundesarbeitsgemeinschaft fu¨r
Rehabilitation e. V., editor. Rehabilitation—Vom Antrag bis zur Nachsorge: fu¨r A¨ rzte, Psychologische
Psychotherapeuten und andere Gesundheitsberufe. Berlin: Springer; 2018. pp. 409–23. 10. Meng K, Holderied A, Vogel H. Rehabilitationsbedarf in der sozialmedizinischen Begutachtung—
Entwicklung und Evaluation eines Entscheidungsalgorithmus. Rehabilitation (Stuttg). 2007; 46:41–9. https://doi.org/10.1055/s-2007-958533 PMID: 17315133. 11. Beck L, Giraud B, Petri B. Tra¨geru¨bergreifende Bedarfsfeststellung—mo¨gliche Ansa¨tze und Perspekti-
ven. Rehabilitation (Stuttg). 2011; 50:11–6. https://doi.org/10.1055/s-0030-1270433 PMID: 21321819. 12. Bethge M, Radoschewski FM, Gutenbrunner C. The Work Ability Index as a screening tool to identify
the need for rehabilitation: longitudinal findings from the Second German Sociomedical Panel of
Employees. J Rehabil Med. 2012; 44:980–7. https://doi.org/10.2340/16501977-1063 PMID: 23027375. 13. Bethge M, Spanier K, Peters E, Michel E, Radoschewski M. Author Contributions Conceptualization: Chloe´ Charlotte Schro¨der, Ju¨rgen Breckenkamp, Jean-Baptist du Prel. Formal analysis: Chloe´ Charlotte Schro¨der, Ju¨rgen Breckenkamp. Methodology: Chloe´ Charlotte Schro¨der, Ju¨rgen Breckenkamp, Jean-Baptist du Prel. Supervision: Ju¨rgen Breckenkamp, Jean-Baptist du Prel. 12 / 16 PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 PLOS ONE Need for and utilization of rehabilitation in older employees with migrant background in Germany Visualization: Chloe´ Charlotte Schro¨der. Writing – original draft: Chloe´ Charlotte Schro¨der. Writing – review & editing: Chloe´ Charlotte Schro¨der, Ju¨rgen Breckenkamp, Jean-Baptist du
Prel. Writing – review & editing: Chloe´ Charlotte Schro¨der, Ju¨rgen Breckenkamp, Jean-Baptist du
Prel. References Self-Reported Work Ability Predicts Reha-
bilitation Measures, Disability Pensions, Other Welfare Benefits, and Work Participation: Longitudinal
Findings from a Sample of German Employees. J Occup Rehabil. 2018; 28:495–503. https://doi.org/
10.1007/s10926-017-9733-y PMID: 28956225 14. Bethge M, Spanier K, Streibelt M. Using Administrative Data to Assess the Risk of Permanent Work
Disability: A Cohort Study. J Occup Rehabil. 2021; 31:376–82. https://doi.org/10.1007/s10926-020-
09926-7 PMID: 32910345. 15. Bethge M, Egner U, Streibelt M, Radoschewski FM, Spyra K. Risikoindex Erwerbsminderungsrente
(RI-EMR). Eine prozessdatenbasierte Fall-Kontroll-Studie mit 8500 Ma¨nnern und 8405 Frauen. Bun-
desgesundheitsblatt—Gesundheitsforschung—Gesundheitsschutz. 2011; 54:1221–8. https://doi.org/
10.1007/s00103-011-1366-2 PMID: 22015794. PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 13 / 16 PLOS ONE Need for and utilization of rehabilitation in older employees with migrant background in Germany 16. Deck R, Tra¨der J-M, Raspe H. Identifikation von potenziellem Reha-Bedarf in der Hausarztpraxis. Idee
und Wirklichkeit. Rehabilitation (Stuttg). 2009; 48:73–83. https://doi.org/10.1055/s-0028-1102952
PMID: 19421938. 17. Deutsche Rentenversicherung Nord, editor. Checkliste fu¨r behandelnde A¨ rzte zur Feststellung von
Rehabilitationsbedarf [Internet]. 2019 [cited 18 Dez 2020]. https://www.deutsche-rentenversicherung. de/SharedDocs/Formulare/DE/Traeger/Nord/K8011.html?groupName_str=formulare 18. Sundstrup E, Hansen ÅM, Mortensen EL, Poulsen OM, Clausen T, Rugulies R, et al. Cumulative occu-
pational mechanical exposures during working life and risk of sickness absence and disability pension. Prospective cohort study. Scand J Work Environ Health. 2017; 43:415–25. https://doi.org/10.5271/
sjweh.3663 PMID: 28783203. 19. Labriola M, Feveile H, Christensen KB, Strøyer J, Lund T. The impact of ergonomic work environment
exposures on the risk of disability pension. Prospective results from DWECS/DREAM. Ergonomics. 2009; 52:1419–22. https://doi.org/10.1080/00140130903067771 PMID: 19851908. 20. Dragano N, Schneider L. Psychosoziale Arbeitsbelastungen als Pra¨diktoren der krankheitsbedingten
Fru¨hberentung: Ein Beitrag zur Beurteilung des Rehabilitationsbedarfs. Rehabilitation (Stuttg). 2011;
50:28–36. https://doi.org/10.1055/s-0030-1270431 PMID: 21321822. 21. Kroll LE, Mu¨ters S, Dragano N. Arbeitsbelastungen und Gesundheit. GBE kompakt; 2(5). Berlin: Robert
Koch-Institut; 2011. https://www.rki.de/DE/Content/Gesundheitsmonitoring/
Gesundheitsberichterstattung/GBEDownloadsK/2011_5_Arbeitsbelastungen.pdf 22. Schro¨der CC, Dyck M, Breckenkamp J, Hasselhorn HM, Du Prel J-B. Utilization of rehabilitation services
for non-migrant and migrant groups of higher working age in Germany—results of the lidA cohort study. BMC Health Serv Res. 2020; 20:31. https://doi.org/10.1186/s12913-019-4845-z PMID: 31924217. 23. Brzoska P, Voigtla¨nder S, Spallek J, Razum O. Arbeitsunfa¨lle, Berufskrankheiten und Erwerbsminder-
ung bei Menschen mit Migrationshintergrund. In: Schott T, Razum O, editors. Migration und medizi-
nische Rehabilitation. Weinheim: Beltz Juventa; 2013. pp. 49–61. 24. Ronda Pe´rez E, Benavides FG, Levecque K, Love JG, Felt E, van Rossem R. Differences in working
conditions and employment arrangements among migrant and non-migrant workers in Europe. Ethnicity
& Health. 2012; 17:563–77. References https://doi.org/10.1080/13557858.2012.730606 PMID: 23534504. 25. Scho¨nfeld S, Schro¨der CC, Du Prel J-B, Razum O, Breckenkamp J. Arbeitsbelastungen und Rehabilita-
tionsbedarf bei a¨lteren Erwerbsta¨tigen mit und ohne Migrationshintergrund—Ergebnisse der lidA
Kohortenstudie. Das Gesundheitswesen. 2021; 83:1–9. Forthcoming. 26. Wahrendorf M. Arbeitsbedingungen im Lebenslauf und Gesundheit im Alter [habilitation thesis]. Du¨s-
seldorf: Heinrich-Heine-Universita¨t Du¨sseldorf; 2019. 27. Klein J, von dem Knesebeck O. Inequalities in health care utilization among migrants and non-migrants
in Germany. A systematic review. Int J Equity Health. 2018; 17:160. https://doi.org/10.1186/s12939-
018-0876-z PMID: 30382861. 28. Eurofound. How your birthplace affects your workplace. Luxembourg: Publications Office of the Euro-
pean Union; 2019. 29. Kuo BCH. Coping, acculturation, and psychological adaptation among migrants: a theoretical and
empirical review and synthesis of the literature. Health Psychology and Behavioral Medicine. 2014;
2:16–33. Epub 2014/01/02. https://doi.org/10.1080/21642850.2013.843459 PMID: 25750766. 30. Spallek J, Razum O. Migration und Gesundheit. In: Richter M, Hurrelmann K, editors. Soziologie von
Gesundheit und Krankheit. Wiesbaden: Springer Fachmedien Wiesbaden; 2016. pp. 153–66. 31. Brzoska P, Voigtla¨nder S, Spallek J, Razum O. Utilization and effectiveness of medical rehabilitation in
foreign nationals residing in Germany. Eur J Epidemiol. 2010; 25:651–60. https://doi.org/10.1007/
s10654-010-9468-y PMID: 20571880. 32. Voigtla¨nder S, Brzoska P, Spallek J, Exner A-K, Razum O. Die Inanspruchnahme medizinischer Reha-
bilitation bei Menschen mit Migrationshintergrund. In: Schott T, Razum O, editors. Migration und medizi-
nische Rehabilitation. Weinheim: Beltz Juventa; 2013. pp. 92–104. 33. Dyck M, Breckenkamp J, Wicherski J, Schro¨der CC, Du Prel J-B, Razum O. Utilization of medical reha-
bilitation services by persons of working age with a migrant background, in comparison to non-migrants:
a scoping review. Public Health Rev. 2020; 41:17. https://doi.org/10.1186/s40985-020-00134-5 PMID:
32774989. 34. Brzoska P, Yilmaz-Aslan Y, Razum O. Zugang und Wirksamkeit bei der medizinischen Rehabilitation
fu¨r Menschen mit Migrationshintergrund. Public Health Forum. 2011; 19:651. https://doi.org/10.1016/j. phf.2011.10.003 35. Schwarz B, Markin K, Salman R, Gutenbrunner C. Barrieren fu¨r Migranten beim Zugang in die medizi-
nische Rehabilitation der gesetzlichen Rentenversicherung. Rehabilitation (Stuttg). 2015; 54:362–8. https://doi.org/10.1055/s-0041-108279 PMID: 26676733. PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 14 / 16 PLOS ONE Need for and utilization of rehabilitation in older employees with migrant background in Germany 36. Hasselhorn HM, Peter R, Rauch A, Schro¨der H, Swart E, Bender S, et al. Cohort profile: the lidA Cohort
Study-a German Cohort Study on Work, Age, Health and Work Participation. Int J Epidemiol. 2014;
43:1736–49. https://doi.org/10.1093/ije/dyu021 PMID: 24618186. 37. Schro¨der H, Kersting A, Gilberg R, Steinwede J. References Methodenbericht zur Haupterhebung lidA-leben in der
Arbeit. FDZ-Methodenreport 01/2013. Nu¨rnberg: Forschungsdatenzentrum der Bundesagentur fu¨r
Arbeit im Institut fu¨r Arbeitsmarkt- und Berufsforschung; 2013. http://doku.iab.de/fdz/reporte/2013/MR_
01-13.pdf. 38. Steinwede J, Kleudgen M, Ha¨ring A, Schro¨der H. Methodenbericht zur Haupterhebung lidA-leben in der
Arbeit, 2. Welle. FDZ-Methodenreport 07/2015. Nu¨rnberg: Forschungsdatenzentrum der Bundesagen-
tur fu¨r Arbeit im Institut fu¨r Arbeitsmarkt- und Berufsforschung; 2015. http://doku.iab.de/fdz/reporte/
2015/MR_07-15.pdf. 39. Steinwede J, Ruiz Marcos J, Kleudgen M. Methodenbericht lidA Welle 3 [Preprint]. Nu¨rnberg: For-
schungsdatenzentrum der Bundesagentur fu¨r Arbeit im Institut fu¨r Arbeitsmarkt- und Berufsforschung;
2018. 40. Nu¨bling M, Andersen HH, Mu¨hlbacher A. Entwicklung eines Verfahrens zur Berechnung der ko¨rperli-
chen und psychischen Summenskalen auf Basis der SOEP-Version des SF 12 (Algorithmus). Data
Documentation No. 16. Berlin: Deutsches Institut fu¨r Wirtschaftforschung; 2006. https://www.diw.de/
documents/publikationen/73/diw_01.c.44987.de/diw_datadoc_2006-016.pdf 41. Ware JE. How to score version 2 of the SF-12 health survey. (with a supplement documenting version
1). Lincoln, R.I., Boston, Mass.: QualityMetric Inc.; Health Assessment Lab; 2005. 42. Ebener M, Hasselhorn HM. Validation of Short Measures of Work Ability for Research and Employee
Surveys. Int J Environ Res Public Health. 2019; 16. https://doi.org/10.3390/ijerph16183386 PMID:
31547466. 43. Siegrist J, Starke D, Chandola T, Godin I, Marmot M, Niedhammer I, et al. The measurement of effort–
reward imbalance at work: European comparisons. Social Science & Medicine. 2004; 58:1483–99. https://doi.org/10.1016/S0277-9536(03)00351-4 PMID: 14759692 44. Siegrist J. Soziale Krisen und Gesundheit. Eine Theorie der Gesundheitsfo¨rderung am Beispiel von
Herz-Kreislauf-Risiken im Erwerbsleben. Go¨ttingen: Hogrefe Verlag; 1996. 45. Schenk L, Bau A-M, Borde T, Butler J, Lampert T, Neuhauser H, et al. Mindestindikatorensatz zur
Erfassung des Migrationsstatus. Empfehlungen fu¨r die epidemiologische Praxis. Bundesgesundheits-
blatt Gesundheitsforschung Gesundheitsschutz. 2006; 49:853–60. https://doi.org/10.1007/s00103-
006-0018-4 PMID: 16927038. 46. Jo¨ckel KH, Babitsch B, Bellach BM et al. Messung und Quantifizierung soziodemographischer Merk-
male in epidemiologischen Studien In: Ahrens W, Bellach BM, Jo¨ckel KH, editors. Messung soziodemo-
graphischer Merkmale in der Epidemiologie. RKI-Schriften 1/1998. Mu¨nchen: MMV Medizin; 1998. p. 7–38. 47. Seaman SR, White IR. Review of inverse probability weighting for dealing with missing data. Stat Meth-
ods Med Res. 2013; 22:278–95. Epub 2011/01/10. https://doi.org/10.1177/0962280210395740 PMID:
21220355. 48. Brzoska P, Sauzet O, Breckenkamp J. Unobserved heterogeneity and the comparison of coefficients
across nested logistic regression models: how to avoid comparing apples and oranges. Int J Public
Health. 2017; 62:517–20. https://doi.org/10.1007/s00038-016-0918-5 PMID: 27812725. 49. Spanier K, Bethge M. Web-based information guide to promote application for medical rehabilitation: a
randomized controlled trial. Journal of quantitative research in rehabilitation medicine. References 2018; 1:21–5. 50. Wenzel T-R, Morfeld M. Nutzung der ICF in der medizinischen Rehabilitation in Deutschland. Anspruch
und Wirklichkeit. Bundesgesundheitsblatt Gesundheitsforschung Gesundheitsschutz. 2017; 60:386–
93. https://doi.org/10.1007/s00103-017-2517-x PMID: 28197665. 51. World Health Organization. International Classification of Functioning, Disability and Health. Geneva:
World Health Organization; 2001 [cited 22 Jan 2021]. http://www.who.int/classifications/icf/en/ 52. Fuchs H. Ermittlung des Rehabilitationsbedarfs–Auswirkungen des Bundesteilhabegesetzes [Internet]. Heidelberg: Deutsche Vereinigung fu¨r Rehabilitation. 2017 [cited 2021 Jan 18]. https://www.reha-recht. de/fachbeitraege/beitrag/artikel/beitrag-d50-2017/ 53. Liu S, Reinhardt JD, Zhang X, Ehrmann C, Cai W, Prodinger B, et al. System-wide Clinical Assessment
of Functioning Based on the International Classification of Functioning, Disability and Health in China:
Interrater Reliability, Convergent, Known Group, and Predictive Validity of the ICF Generic-6. Arch
Phys Med Rehabil. 2019; 100:1450–1457.e1. Epub 2018/12/14. https://doi.org/10.1016/j.apmr.2018. 11.014 PMID: 30557550. 54. Stucki G, Ewert T, Cieza A. Value and application of the ICF in rehabilitation medicine. Disability and
Rehabilitation. 2002; 24:932–8. https://doi.org/10.1080/09638280210148594 PMID: 12523361. PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 15 / 16 PLOS ONE Need for and utilization of rehabilitation in older employees with migrant background in Germany 55. Prodinger B, Stucki G, Coenen M, Tennant A. The measurement of functioning using the International
Classification of Functioning, Disability and Health: comparing qualifier ratings with existing health sta-
tus instruments. Disability and Rehabilitation. 2019; 41:541–8. Epub 2017/10/08. https://doi.org/10. 1080/09638288.2017.1381186 PMID: 28988490. 55. Prodinger B, Stucki G, Coenen M, Tennant A. The measurement of functioning using the International
Classification of Functioning, Disability and Health: comparing qualifier ratings with existing health sta-
tus instruments. Disability and Rehabilitation. 2019; 41:541–8. Epub 2017/10/08. https://doi.org/10. 1080/09638288.2017.1381186 PMID: 28988490. 56. Ehrmann C, Prodinger B, Stucki G, Cai W, Zhang X, Liu S, et al. ICF Generic Set as new standard for
the system wide assessment of functioning in China: a multicentre prospective study on metric proper-
ties and responsiveness applying item response theory. BMJ Open. 2018; 8:e021696. Epub 2018/12/
14. https://doi.org/10.1136/bmjopen-2018-021696 PMID: 30552245. 57. Prodinger B, Reinhardt JD, Selb M, Stucki G, Yan T, Zhang X, et al. Towards system-wide implementa-
tion of the International Classification of Functioning, Disability and Health (ICF) in routine practice:
Developing simple, intuitive descriptions of ICF categories in the ICF Generic and Rehabilitation Set. J
Rehabil Med. 2016; 48:508–14. https://doi.org/10.2340/16501977-2066 PMID: 27008067. 58. Schlo¨ffel M, Kampling H, Fichtner U, Farin-Glattacker E, Pollmann H, Mittag O. PLOS ONE | https://doi.org/10.1371/journal.pone.0263643
February 7, 2022 References Online-Rehabedarfstest
(OREST): Wirksamkeit einer Einladung zu einem proaktiven Screening (Selbsttest) auf Bedarf an medi-
zinischen Rehabilitationsmaßnahmen bei Versicherten der Deutschen Rentenversicherung Baden-
Wu¨rttemberg und Rheinland. Rehabilitation. 2020. Epub 2020/11/05. https://doi.org/10.1055/a-1282-
8564 PMID: 33152781. 59. Spanier K, Streibelt M, U¨ nalan F, Bethge M. A web-based intervention to promote applications for reha-
bilitation: a study protocol for a randomized controlled trial. Trials. 2015; 16:436. Epub 2015/09/29. https://doi.org/10.1186/s13063-015-0968-7 PMID: 26420450. 16 / 16
|
https://openalex.org/W2522253462
|
http://www.jeeng.net/pdf-64505-5127?filename=RELIABILITY OF A.pdf
|
English
| null |
RELIABILITY OF A COLLECTIVE WASTEWATER TREATMENT PLANT
|
Journal of Ecological Engineering
| 2,016
|
cc-by
| 2,763
|
ABSTRACT The paper discusses technological reliability of a collective wastewater treatment plant
(WTP). The WTP capacity expressed as population equivalents (PE) was 22,500. The
research was conducted in 2013 and 2014 and it involved the collection and physical
and chemical analysis of 101 samples of raw and treated sewage. Reliability analysis
was based on Weibull reliability theory. Reliability of retaining organic compounds
defined as BOD5 was 68%, and for biogenic compounds, such as total nitrogen and
total phosphorus, it was 92% and 88%, respectively. Keywords: sewage, wastewater treatment plant, technological reliability, Weibull
theory INTRODUCTION reducing the operating costs of sewerage systems
[Skoczko 2012]. In these WTPs, the quality of
both supplied and discharged sewage is not regu
larly monitored and this is why the operators do
not have enough information on the sewage qual
ity. This study discusses the possibility of using
Weibull reliability method for determination of
wastewater treatment plant operational reliability. As showed in previous works, Weibull distribu
tion is a useful, general probability distribution
applicable in reliability assessment [Bugajski et
al. 2012, Bugajski 2014]. According to the binding Regulation of the
Minister of Environment as of 18 November [J. of Laws as of 16 December 2014], accuracy of
wastewater treatment plant operation is based
on determination of pollution concentrations in
treated sewage that is then compared with accept
able concentrations set out in this Regulation. The
Regulation provides also a number of samples in
which the limit values may be exceeded without
negatively affecting the facility performance. Therefore, if pollution concentrations for a spe
cific number of samples are known, unequivo
cal conclusions on the reliability of a wastewater
treatment plant may be drawn. However, a WTP
operator also needs to know technological reli
ability of a facility, that is its capacity for proper
treatment of specific amounts of sewage [Mu
cha and Mikosz 2009, Krzanowski and Wałęga
2006, Andraka 1997a, Andraka 1997b]. This is
particularly important in the WTPs that do not
monitor the quality of the sewage discharged into
a receiver on a regular basis [Kuśnierz and Zar
zycka 2014], i.e. in a majority of small and me
dium WTPs located in the rural areas of Poland. This lack of continuous monitoring is caused by Journal of Ecological Engineering
Volume 17, Issue 4, Sept. 2016, pages 143–147
DOI: 10.12911/22998993/64505 Journal of Ecological Engineering
Volume 17, Issue 4, Sept. 2016, pages 143–147
DOI: 10.12911/22998993/64505 Research Article Piotr Bugajski1, Krzysztof Chmielowski1, Grzegorz Kaczor1 Piotr Bugajski1, Krzysztof Chmielowski1, Grzegorz Kaczor1 1 Department of Sanitary Engineering and Water Management, Faculty of Environmental Engineering and
Land Surveying, H. Kołłątaja University of Agriculture in Kraków, Al. Mickiewicza 24/28, 30-059 Kraków,
Poland, e-mail: p.bugajski@ur.krakow.pl Received: 2016.05.13
Accepted: 2016.08.07
Published: 2016.09.30 RESULT ANALYSIS The calcula
tions were used to prepare a histogram presenting
individual BOD5 values, as showed in Figure 1. Chemical analysis indicated that in 72 out of 101
samples of treated sewage BOD5 ranged from 10
to 15 mgO2⋅dm-3. Percentage share of BOD5 in
this interval was nearly 71%, as compared with
all analyzed sewage samples. The study revealed
nine cases of exceeding the acceptable BOD5 𝑓𝑓(𝑥𝑥) = 𝑐𝑐
𝑏𝑏∙(𝑥𝑥−Ө
𝑏𝑏
)
(𝑐𝑐−1)
∙𝑒𝑒−(𝑥𝑥−Ө
𝑏𝑏)
𝑐𝑐
(1) (1) where: x – variable indicating the concentration
of a specific pollution indicator in the
treated sewage, b – scale parameter,
c – shape parameter,
θ – location parameter. Assumptions: θ < x, b > 0, c > 0 b – scale parameter, c – shape parameter, θ – location parameter. Assumptions: θ < x, b > 0, c > 0 Assumptions: θ < x, b > 0, c > 0 Weibull distribution parameters were esti
mated by means of maximum likelihood method. The applicability of Weibull distribution to the
empirical data was assessed using Hollander-
Proschan test. The results were analyzed with
STATISTICA 8 software. Weibull distribution parameters were esti
mated by means of maximum likelihood method. y
The applicability of Weibull distribution to the
empirical data was assessed using Hollander-
Proschan test. The results were analyzed with
STATISTICA 8 software. When the class boundaries were established, the
values within individual class intervals were com
puted. These activities yielded a series in which
each interval comprised an assigned number of
observations called class abundance. The calcula
tions were used to prepare a histogram presenting
individual BOD5 values, as showed in Figure 1. Chemical analysis indicated that in 72 out of 101
samples of treated sewage BOD5 ranged from 10
to 15 mgO2⋅dm-3. Percentage share of BOD5 in
this interval was nearly 71%, as compared with
all analyzed sewage samples. The study revealed
nine cases of exceeding the acceptable BOD5 Mean designed daily sewage supply for the
WTP in Biłgoraj was Qms= 8000 m3·d-1, and
maximum supply was Qdmax.= 10,000 m3·d-1. The
facility size according to the population equiva
lent (PE) ranged between 15,000 and 99,000 PE. Mean daily sewage supply in the study period
was 4000–5000 m3·d-1. STUDY SCOPE AND OBJECTIVES The aim of the study was to use Weibull meth
od to determine the reliability of organic and bio
genic compound removal in a collective wastewa
ter treatment plant. Organic pollution was defined
as BOD5 and biogenic pollution was calculated
based on total nitrogen and total phosphorus. The
study included only those pollution indicators in
the treated sewage for which limit values set out
in the water permit for the facility were exceeded. The study covered the period of two years and
was conducted in a collective wastewater treat 143 Journal of Ecological Engineering Vol. 17(4), 2016 RESULT ANALYSIS ment plant in Biłgoraj. A total of 101 samples of
sewage were collected and subjected to physical
and chemical analyses. The efficiency of pollu
tion removal in the facility was determined by
employing some elements of Weibull reliability
theory. Weibull distribution is characterized by a
probability density function (1) with parameters
b, c and θ:𝑓𝑥𝑐𝑥𝑐𝑒𝑥𝑐 The performance of the analyzed WTP was
assessed in two stages. At the first stage, the fre
quency of specific volumes and concentrations
of the analyzed parameters was determined. The
analysis of the frequency of specific volumes
and the concentrations are very useful statisti
cal tools for the interpretation of data collected
at irregular time intervals. Following calcula
tions of frequency for the volume/concentration
of individual indicators, the number of classes for
each frequency distribution was determined. Ten
classes were defined for BOD5 and the range of
class interval was 5 mgO2⋅dm-3. For total nitro
gen, there were 11 classes with the range of class
interval 1 mgN⋅dm-3, and for total phosphorus
there were eight classes with the range of class
interval 0.4 mgP⋅dm-3. The class intervals were
determined in such a way that the frequency dis
tribution would yield as detailed and clear image
of the statistical collection structure as possible. The performance of the analyzed WTP was
assessed in two stages. At the first stage, the fre
quency of specific volumes and concentrations
of the analyzed parameters was determined. The
analysis of the frequency of specific volumes
and the concentrations are very useful statisti
cal tools for the interpretation of data collected
at irregular time intervals. Following calcula
tions of frequency for the volume/concentration
of individual indicators, the number of classes for
each frequency distribution was determined. Ten
classes were defined for BOD5 and the range of
class interval was 5 mgO2⋅dm-3. For total nitro
gen, there were 11 classes with the range of class
interval 1 mgN⋅dm-3, and for total phosphorus
there were eight classes with the range of class
interval 0.4 mgP⋅dm-3. The class intervals were
determined in such a way that the frequency dis
tribution would yield as detailed and clear image
of the statistical collection structure as possible. When the class boundaries were established, the
values within individual class intervals were com
puted. These activities yielded a series in which
each interval comprised an assigned number of
observations called class abundance. RESULT ANALYSIS As per the water permit
issued in 2004, acceptable concentrations of the
analyzed parameters in the sewage discharged
into a receiver should not exceed 15 mgO2·dm-3
for BOD5, 15 mgN·dm-3 for total nitrogen and
2 mgP·dm-3w for total phosphorus. Figure 1. Histogram of characteristic BOD5 values in treated sewage Figure 1. Histogram of characteristic BOD5 values in treated sewage 144 Journal of Ecological Engineering Vol. 17(4), 2016 limit that for this facility was 15 mgO2⋅dm-3. Total
percentage share of BOD5 values exceeding the
acceptable limit for all class intervals was 8.9%. The determined distribution parameters were
used to verify a hypothesis assuming the useful
ness of Weibull distribution for the approximation
of empirical data. Statistical analysis of the prob
ability test p for all the indicators confirmed that
the empirical data may be described by Weibull
distribution and assumed as a null hypothesis. Results of distribution fit based on Hollander-
Proschan test together with estimated parameters
are presented in Table 1. Concentrations of total nitrogen in the sam
ples of treated sewage fell into two distinct in
tervals, i.e. from 13 to 14 mgN·dm-3 and from
14 to 15 mgNog·dm-3. Total percentage share of
total nitrogen in these intervals was 86%. Ac
ceptable concentration limit of total nitrogen was
exceeded in only one sample. This is less than
1% of all samples. Characteristic concentrations
of total nitrogen in individual class intervals are
presented in Figure 2. The reliability analysis based on Weibull
method was used to determine the probability of
occurrence of the value/concentration higher than
that provided in the water permit for the facility
for all three parameters. The analysis of the second eutrophication in
dicator, i.e. total phosphorus, revealed no domi
nant intervals of its characteristic concentrations
in the treated sewage. Majority of concentra
tion values (93 out of 101) fell within 0.4 to 2.0
mg P·dm-3 range. Total percentage share of total
phosphorus concentrations in these intervals was
92%. In five samples, the concentration of total
nitrogen was higher than the acceptable limit of
2.0 mg P·dm-3. Total percentage share of total
phosphorus in 2.0 to 3.2 intervals was 4.95%. Characteristic concentrations of total phosphorus
in the treated sewage are presented in Figure 3. Cumulative distribution for BOD5 showed
in Figure 4 indicated that the probability of ex
ceeding the acceptable limit of 15 mgO2·dm-3
was 42%. RESULT ANALYSIS Therefore, acceptable concentration
of BOD5 in the sewage discharged into the re
ceiver was exceeded on about 153 days per year. The guidelines prepared by Andraka and Dzienis
[2003] suggested that the reliability of a waste
water treatment plant of this size should be at
least 92.3%, for the manufacturer’s risk α=0.05. These guidelines allow ineffective operation of a
WTP with PE ranging from 15,000 to 49,999 for
27 days per year. A comparison of the number of
days in which BOD5 was higher than acceptable Cumulative distribution for BOD5 showed
in Figure 4 indicated that the probability of ex
ceeding the acceptable limit of 15 mgO2·dm-3
was 42%. Therefore, acceptable concentration
of BOD5 in the sewage discharged into the re
ceiver was exceeded on about 153 days per year. The second stage of the study focused on re
liability analysis of the investigated facility and
took into account three indicators of pollution. Figure 2. Histogram of characteristic concentrations of total nitrogen in treated sewage Figure 2. Histogram of characteristic concentrations of total nitrogen in treated sewage Table 1. Results of estimation of Weibull distribution parameters with goodness of fit to empirical data
Parameter
Distribution parameters
Hollander-Proschan test
b
c
θ
Test value
p
BOD5
16.337
1.8815
4.1016
0.551925
0.58100
Total nitrogen
14.230
17.917
-1.000
-1.22547
0.22040
Total phosphorus
1.4769
2.4871
0.21515
0.223653
0.82303 Table 1. Results of estimation of Weibull distribution parameters with goodness of fit to empirical data 145 Journal of Ecological Engineering Vol. 17(4), 2016 Journal of Ecological Engineering Vol. 17(4), 2016 Figure 3. Histogram of characteristic concentrations of total phosphorus in treated sewage Figure 3. Histogram of characteristic concentrations of total phosphorus in treated sewage Figure 5. Results of Weibull reliability analysis for
total nitrogen in treated sewage
Figure 6. Results of Weibull reliability analysis for
total phosphorus in treated sewage Figure 4. Results of Weibull reliability analysis for
BOD5 in treated sewage Figure 5. Results of Weibull reliability analysis for
total nitrogen in treated sewage Figure 4. Results of Weibull reliability analysis for
BOD5 in treated sewage Figure 5. Results of Weibull reliability analysis for
total nitrogen in treated sewage Figure 4. Results of Weibull reliability analysis for
BOD5 in treated sewage Figure 6. Results of Weibull reliability analysis for
total phosphorus in treated sewage with the number of days in which exceeding this
value does not affect the facility performance
revealed that the normal range for BOD5 in the
discharged sewage was exceeded for 126 days
per year. However, significant exceedance, i.e. above 40 mgO2⋅dm-3 was noticed in only 5% of
all cases. When only those significant excess
es were considered, Weibull reliability theory
showed that the efficiency of the facility was as
high as 90%. Reliability analysis of total nitrogen remov
al revealed the probability of its concentration
exceeding the acceptable limit was about 8%. Therefore, nitrogen concentrations higher than
the acceptable 15 mgN·dm-3 could be expected on
29 days per year. This means the effectiveness of
removing nitrogen compounds in this facility, de
fined here as total nitrogen, was 92%. The results
of Weibull reliability method for total nitrogen in
the treated sewage are presented in Figure 5. Figure 6. Results of Weibull reliability analysis for
total phosphorus in treated sewage Acceptable limits of total phosphorus
(2 mgP·dm-3), the second biogenic indicator, in
the treated sewage were exceeded in 12% of the
analyzed cases, as showed in Figure 6. Therefore,
the concentration of total phosphorus was likely 146 Journal of Ecological Engineering Vol. 17(4), 2016 o/Kraków, Zakopane. to be above the acceptable limit on 44 days per
year. This means that reliability of total phospho
rus removal in this facility was 88%. 2. Andraka D. 1997b. Prognozowanie niezawodności
oczyszczalni ścieków na przykładzie miejskiej oc
zyszczalni w Grajewie. Mat. IX Ogólnopolskiej
Konferencji Naukowo-Technicznej nt. „Problemy
gospodarki wodno-ściekowej w regionach rolniczo-
przemysłowych”, Rajgród 16–24 czerwca, 366–373. CONCLUSIONS 3. Andraka D., Dzienis L. 2003. Wymagany poziom
niezawodności oczyszczalni ścieków w świetle
przepisów polskich i europejskich. Zesz. Nauk. Politechniki
Białostockiej,
Seria
Inżynieria
Środowiska, z. 16, Białystok, 24–28. 1. Reliability of the investigated wastewater
treatment plant to remove organic and eutro
phic pollutants defined as BOD5, total nitrogen
and total phosphorus was periodically limited. 4. Bugajski P. 2014. Ocena niezawodności usuwania
związków biogennych w oczyszczalni ścieków
metodą Weibulla. Zeszyty Problemowe Postępów
Nauk Rolniczych nr 276, 13–21. 2. Reliability of retaining organic compounds de
fined as BOD5 was 68%, and for biogenic com
pounds such as total nitrogen and total phos
phorus it was 92% and 88%, respectively. 5. Bugajski P., Wałęga A., Kaczor G. 2012. Zastoso
wanie metody Weibulla do analizy niezawodności
działania przydomowej oczyszczalni ścieków. Gaz, Woda i Technika Sanitarna 2, 56–58. 3. Assessment of the general reliability of the
facility as per Weibull reliability model
demonstrated its effective operation for 239
days per year for BOD5, 336 days per year
for total phosphorus and 321 days per year
for total nitrogen. 6. Mucha Z., Mikosz J. 2009. Racjonalne stosowanie
małych oczyszczalni ścieków z uwzględnieniem
kryteriów zrównoważonego rozwoju. Czasopismo
Techniczne, 2, 91–100. 4. With view of the above, the processes of ni
trification and denitrification in a biologi
cal reactor of the investigated facility should
be streamlined to provide more effective re
moval of organic and nitrogen-based com
pounds. Moreover, the process of chemi
cal precipitation of phosphorus compounds
should also be improved. 7. Krzanowski S., Wałęga A. 2006. Wykorzysta
nie teorii niezawodności i statystycznej kontroli
jakości do oceny eksploatacyjnej wiejskich oc
zyszczalnie ścieków. Infrastr. Ekol. Ter. Wiej.,
3/2, 17–37. 8. Kuśnierz M., Zarzycka A. 2014. Zakres i częstotli
wość prowadzenia badań i pomiarów w racjonal
nej eksploatacji oczyszczalni ścieków. Inżynieria i
Ochrona Środowiska, 17(1), 63–73. 9. Rozporządzenie Ministra Środowiska z dnia 18 li
stopada 2014 r. w sprawie warunków jakim powin
ny odpowiadać ścieki wprowadzane do wód lub do
ziemi, oraz w sprawie substancji szczególnie szko
dliwych dla środowiska wodnego (Dz. U. z dnia 16
grudnia 2014 r., poz. 1800). Pracę dofinansowano ze środków Wojewódzkiego Funduszu Ochrony Środowiska
i Gospodarki Wodnej w Lublinie. REFERENCES 1. Andraka D. 1997a. Ocena i prognozowanie
niezawodności
oczyszczalni
ścieków
na
przykładzie oczyszczalni w Białymstoku i Gra
jewie. Ogólnopolska Konferencja Naukowo-
Techniczna nt. Bezpieczeństwo i niezawodność
działania systemów gazowych, wodociągowych,
kanalizacyjnych i centralnego ogrzewania, PZiTS 10. Skoczko I. 2012. Ekonomiczna analiza rozwią
zań oczyszczalni ścieków dla wsi przy rozproszo
nej zabudowie. Gaz, Woda i Technika Sanitarna
nr 6, 271–275. 147 147
|
https://openalex.org/W3185193907
|
https://cronfa.swan.ac.uk/Record/cronfa57411/Download/57411__20616__263eda6a4cc34670a9f86686f09d621f.pdf
|
English
| null |
Robust topological designs for extreme metamaterial micro-structures
|
Scientific reports
| 2,021
|
cc-by
| 12,479
|
Robust topological designs
for extreme metamaterial
micro‑structures
OPEN Tanmoy Chatterjee1*, Souvik Chakraborty2, Somdatta Goswami3, Sondipon Adhikari1 &
Michael I. Friswell1 We demonstrate that the consideration of material uncertainty can dramatically impact the optimal
topological micro-structural configuration of mechanical metamaterials. The robust optimization
problem is formulated in such a way that it facilitates the emergence of extreme mechanical
properties of metamaterials. The algorithm is based on the bi-directional evolutionary topology
optimization and energy-based homogenization approach. To simulate additive manufacturing
uncertainty, combinations of spatial variation of the elastic modulus and/or, parametric variation
of the Poisson’s ratio at the unit cell level are considered. Computationally parallel Monte Carlo
simulations are performed to quantify the effect of input material uncertainty to the mechanical
properties of interest. Results are shown for four configurations of extreme mechanical properties: (1)
maximum bulk modulus (2) maximum shear modulus (3) minimum negative Poisson’s ratio (auxetic
metamaterial) and (4) maximum equivalent elastic modulus. The study illustrates the importance
of considering uncertainty for topology optimization of metamaterials with extreme mechanical
performance. The results reveal that robust design leads to improvement in terms of (1) optimal mean
performance (2) least sensitive design, and (3) elastic properties of the metamaterials compared to
the corresponding deterministic design. Many interesting topological patterns have been obtained for
guiding the extreme material robust design. Metastructures are metamaterial induced concepts implanted in structural design. The exploration of metama-
terials was heavily steered by the path-breaking investigation of electromagnetic metamaterials, which exhibited
properties like, negative permittivity and permeability1. This paved the way for mechanical metamaterials2. These metamaterials are engineered to derive their fundamental mechanical properties from the geometry of
their structural building blocks (periodicity or translational symmetry), instead of the constituting materials3,4. Mechanical metamaterials have gained wide popularity in engineering applications due to their superior proper-
ties compared to conventional materials found in nature. It has been found that by varying one or more material
constants such as, bulk modulus, shear modulus, Poisson’s ratio and elastic modulus, extraordinary materials
such as, ultra light weight and high strength aerospace shells, functionally graded electromagnetic sensors and
energy absorbing dampers can be secured5,6. For exploring such exciting possibilities of discovering new materials with extreme properties, determining
their optimal configuration at the very conceptual micro-structural design stage is indispensable7. A recent
works with multi-material lattices8, pre-stressed materials9 and piezo-embedded damped lattice metamaterials10,11
show some ways of achieving extreme homogeneous properties. www.nature.com/scientificreports www.nature.com/scientificreports Scientific Reports | (2021) 11:15221 Robust topological designs
for extreme metamaterial
micro‑structures
OPEN In this scenario, topology optimization (TO) is
well-known to be an effective computational analysis tool which renders greater design freedom and yields the
best material layout given a prescribed design domain and boundary condition12. Depending on the algorithm’s
performance and versatility, various successful TO methods have emerged to the present13, few popular ones
being the solid isotropic material with penalization (SIMP), evolutionary structural optimization (ESO), level
set method (LSM), and moving morphable components (MMC). In fact, the popularity of TO has even led to its
integration with relatively newer numerical discretization schemes (compared to finite element method) such
as, isogeometric analysis14. A simple illustration of multi-scale design and topology optimization of the micro-
structure of metamaterial is presented in Fig. 1a and b, respectively. 1College of Engineering, Swansea University, Bay Campus, Swansea SA1 8EN, UK. 2Department of Applied
Mechanics, Indian Institute of Technology Delhi, Hauz Khas 110016, India. 3Department of Applied Mathematics,
Brown University, Providence, RI 02912, USA. *email: tanmoy.chatterjee@swansea.ac.uk | https://doi.org/10.1038/s41598-021-94520-x Scientific Reports | (2021) 11:15221 www.nature.com/scientificreports/ www.nature.com/scientificreports/ Figure 1. An illustrative visual tool guide of the key aspects of the proposed design paradigm. The focus is on
optimizing the topology of the periodic unit cell as shown in (a) and determine robust micro-structural designs
against uncertainties. Note that the objective of the deterministic topological design in (b) is to maximize
the bulk modulus (refer Table 1 for details). The negative ordinate indicates the maximization problem. The
3D computer aided design (CAD) geometries of the robust micro-structural configurations shown in (c) are
obtained using Autodesk Fusion 360 software (version 2.0.9930) (https://www.autodesk.co.uk/campaigns/educa
tion/fusion-360). These geometries can be directly exported for 3D printing. The four different geometries
shown here correspond to robust topologies exhibiting extreme mechanical properties (for example, maximum
bulk modulus, maximum shear modulus, minimum Poisson’s ratio and maximum equivalent elastic modulus). Figure 1. An illustrative visual tool guide of the key aspects of the proposed design paradigm. The focus is on
optimizing the topology of the periodic unit cell as shown in (a) and determine robust micro-structural designs
against uncertainties. Note that the objective of the deterministic topological design in (b) is to maximize
the bulk modulus (refer Table 1 for details). The negative ordinate indicates the maximization problem. The
3D computer aided design (CAD) geometries of the robust micro-structural configurations shown in (c) are
obtained using Autodesk Fusion 360 software (version 2.0.9930) (https://www.autodesk.co.uk/campaigns/educa
tion/fusion-360). www.nature.com/scientificreports/ www.nature.com/scientificreports/ The success of TO is clearly evident from its extensive applications for material micro-structural design in
recent years. In this regard, few note-worthy literatures have been discussed as follows. Gibiansky and Sigmund15
investigated multi-phased elastic composites and obtained topological design corresponding to extreme bulk
and shear modulus. For obtaining maximum bulk and shear modulus, the optimal topology of cellular micro-
structures was determined using ESO16. A compact code in MATLAB based on the SIMP and energy-based
homogenization method (EBHM) was developed for extreme material multi-scale structural design17. Clausen
et al.18 designed and fabricated 3D auxetic material micro-structures undergoing large deformations. Long
et al.19 performed topology optimization to maximize the effective Young’s modulus, so as to obtain the optimal
distribution of 3D material micro-structures whose constituent phases consist of non-identical Poisson’s ratios. A novel TO method was presented based on the independent point-wise density interpolation to obtain a bi-
material chiral metamaterial20. Chen and Huang21 designed 3D chiral metamaterials based on the couple-stress
homogenization and maximized the chiralty (the coupling effects between tension/compression and torsion)
of the cellular structure. Xu et al.14 achieved designs of ultra-lightweight architected materials along with extre-
mal properties such as, maximum bulk and shear modulus by using isogeometric TO. Petal-shaped auxetic
metamaterials were designed where a back-propagation neural network was coupled with isogeometric TO to
evaluate their macroscopic equivalent properties22. Zheng et al.23 proposed two numerical schemes within the
evolutionary optimization framework to obtain 2D auxetic metamaterials with favourable characteristics, and
experimentally validated their special properties. Ye et al.24 proposed a method for the simultaneous adaptive
design of gradually stiffer mechanical metamaterials along with auxetic property. f
Although the above published articles related to micro-structural TO explore multiple potent research pros-
pects to achieve extreme material properties, they are restricted to the deterministic optimization framework. In
this regard, the literature is found to be scarce in studies addressing uncertainties in manufacturing processes. The following few works highlight the importance of considering manufacturing variability and their effect on
the structural design. Most nano, micro and macro structures fabricated using e-beam lithography, etching and
milling processes, respectively are vulnerable to manufacturing uncertainties. Even the structures meticulously
designed and optimized may experience inferior performance or in an extreme scenario, lose their functional-
ity due to the wear of machining tools, under or over-etching, or malcalibrated e-beam equipment. www.nature.com/scientificreports/ To address
few of the above aspects, robust topology optimization (RTO) for structural design was presented accounting
for manufacturing errors in25–27. Specifically, the effect of over-etching (erosion) and under-etching (dilation)
was modelled for structures produced by milling or etching, which may cause the structural parts to become
thinner or thicker than intended. For the works25,26, the problem was posed as a worst case design formulation
and the error effects were simulated by a Heaviside projection threshold. While they considered only uniform
manufacturing errors (i.e. constant in magnitude over the entire design domain), the method was extended to
account for non-uniform manufacturing errors (i.e. with spatially varying magnitude)27. For doing so, a proba-
bilistic approach was followed where the projection threshold was represented by a (non-Gaussian) random field. Jansen et al.28 developed an RTO framework which incorporated the spatially varying geometric imperfections
due to misalignment and misplacement of material often encountered in slender (civil engineering) structures. These imperfections should be accounted for in accordance to the provision recommended in the Eurocode for
design of steel structures and by the Joint Committee on Structural Safety. Note that the misplacement errors
cause a perturbation in the spatial location of the material, whereas the errors due to etching, add or remove
material at the structural surface. An ESO based RTO framework was developed for multi-material structures
considering interval loading uncertainty in29. Efficiency was enhanced by (1) orthogonal decomposition of the
loading uncertainty which decoupled the stochastic problem from the optimization loop and (2) sensitivity
analysis. Material uncertainty modelled as imprecise probability was integrated with the multi-scale concurrent
ESO based TO framework in30. The RTO methodology adopted the type I hybrid random interval model. An
ESO based RTO framework was developed for multiscale 2D and 3D structures considering multiple random
loading conditions in31. Decoupled sensitivity analysis was observed to improve the computational efficiency. An
ESO based RTO methodology was proposed for actuator-coupled structures considering hybrid uncertainties
in32. The hybrid random interval model simulated the stochastic mechanical and piezoelectric parameters and
perturbation analysis was employed to evaluate the statistical quantities.f p
y
p y
q
While the above works simulated manufacturing uncertainties numerically and investigated their effect on
the structural topology, the following studies have carried out experimental analysis to probe manufacturing
uncertainties. Robust topological designs
for extreme metamaterial
micro‑structures
OPEN These geometries can be directly exported for 3D printing. The four different geometries
shown here correspond to robust topologies exhibiting extreme mechanical properties (for example, maximum
bulk modulus, maximum shear modulus, minimum Poisson’s ratio and maximum equivalent elastic modulus). Figure 1. An illustrative visual tool guide of the key aspects of the proposed design paradigm. The focus is on
optimizing the topology of the periodic unit cell as shown in (a) and determine robust micro-structural designs
against uncertainties. Note that the objective of the deterministic topological design in (b) is to maximize
the bulk modulus (refer Table 1 for details). The negative ordinate indicates the maximization problem. The
3D computer aided design (CAD) geometries of the robust micro-structural configurations shown in (c) are
obtained using Autodesk Fusion 360 software (version 2.0.9930) (https://www.autodesk.co.uk/campaigns/educa
tion/fusion-360). These geometries can be directly exported for 3D printing. The four different geometries
shown here correspond to robust topologies exhibiting extreme mechanical properties (for example, maximum
bulk modulus, maximum shear modulus, minimum Poisson’s ratio and maximum equivalent elastic modulus). https://doi.org/10.1038/s41598-021-94520-x Scientific Reports | (2021) 11:15221 | www.nature.com/scientificreports/ It has been shown that the emission rate of a 3D printer based on fused deposition modeling prin-
ciple increased significantly with the increase in the extrusion temperature33. Melenka et al.34 found that the per
cent infill has a significant effect on the longitudinal elastic modulus and ultimate strength of the test specimens,
whereas print orientation and layer thickness fail to achieve significance. Dimensional analysis of test specimens
also showed that the test specimen varied significantly from the nominal print dimensions. The manufacturing
tolerances of metamaterial samples produced by a selective laser sintering process were obtained to simulate
the manufacturing uncertainty in mass-produced industrial applications35. It was found that even small levels of
variability, given by less than 1 % for the mass and less than 3 % for the elastic modulus, has a significant effect
on the overall vibration attenuation performance.i While the first of the above three paragraphs discusses works on deterministic TO for extreme metamate-
rial design, the second paragraph illustrates the numerical simulation of manufacturing uncertainty and robust
topological design. Finally, the above paragraph highlights a few instances of experimental exploration of manu-
facturing uncertainties. Motivated by the need to consider manufacturing uncertainties for metamaterial design,
this work attempts to bridge the above points. Thus, we integrate these aspects to present an RTO formulation
specifically for extreme metamaterial design by minimizing the effect of potential material uncertainties. To the
best of the authors’ knowledge, this is one of the first applications of robust micro-structural design for obtain-
ing extreme mechanical properties of metamaterials. Few instances of ready-to-print robust micro-structural Scientific Reports | (2021) 11:15221 | https://doi.org/10.1038/s41598-021-94520-x www.nature.com/scientificreports/ Figure 2. Flowchart of the proposed framework. The three modules, namely, (1) user-defined initialization,
(2) topology optimization under uncertainty analysis and (3) post-processing unit for additive manufacturing,
are bordered by dotted lines which constitute the approach. A square-shaped void region at the centre of the
150 × 150 unit cell is adopted as the initial design (as shown in the right side of the user-defined module block). The robust optimal topology obtained (corresponding to maximum shear modulus subjected to the spatial
variability of the elastic modulus (refer Table 2)) is shown as a 20 × 30 periodic arrangement of the unit cell in
the post-processing module. The 3D fabrication-ready object is obtained after extruding and material rendering
in Autodesk Fusion 360 software (version 2.0.9930) (https://www.autodesk.co.uk/campaigns/education/fusion-
360). www.nature.com/scientificreports/ The abbreviations RTO and UQ represent robust topology optimization and uncertainty quantification,
respectively. Figure 2. Flowchart of the proposed framework. The three modules, namely, (1) user-defined initialization,
(2) topology optimization under uncertainty analysis and (3) post-processing unit for additive manufacturing,
are bordered by dotted lines which constitute the approach. A square-shaped void region at the centre of the
150 × 150 unit cell is adopted as the initial design (as shown in the right side of the user-defined module block). The robust optimal topology obtained (corresponding to maximum shear modulus subjected to the spatial
variability of the elastic modulus (refer Table 2)) is shown as a 20 × 30 periodic arrangement of the unit cell in
the post-processing module. The 3D fabrication-ready object is obtained after extruding and material rendering
in Autodesk Fusion 360 software (version 2.0.9930) (https://www.autodesk.co.uk/campaigns/education/fusion-
360). The abbreviations RTO and UQ represent robust topology optimization and uncertainty quantification,
respectively. topologies of extreme metamaterials achieved as a part of this study have been presented in Fig. 1c for illustrative
purpose and the interest of readers. Periodic boundary conditions.
The displacements on a pair of opposite boundaries of a 2-D base cell Periodic boundary conditions. The displacements on a pair of opposite boundaries of a 2-D base cell Periodic boundary conditions. The displacements on a pair of opposite boundaries of a 2-D base cell (6)
uk−
i
= ε0
ijyk−
j
+ u∗
i
uk+
i
= ε0
ijyk+
j
+ u∗
i (6) where notations k− and k+ represent the pair of two opposite parallel boundary surfaces perpendicular to the
kth direction. One can remove the unknown periodic fluctuation term u∗
i by using the difference between the
displacements as, (7)
uk+
i
−uk−
i
= ε0
ij(yk+
j
−yk−
j
) (7) For a given unit cell, the quantity (yk+
j
−yk−
j
) is constant. Thus, the right side of Eq. (7) is a constant for a
specified ε0
ij . This boundary conditions can be incorporated within the finite element model by restraining the
corresponding pairs of nodal displacements. Note that this boundary configuration satisfies the periodicity and
continuity conditions in terms of displacement and stress employing displacement-based finite element analysis37. Optimization approach. The unit cell is discretized into N finite elements, resulting into equal number
of density design variables ρ ∈RN . The element level elastic modulus can be defined by the modified SIMP
method38 as (8)
Ee(ρe) = Emin + ρp
e (E0 −Emin) (8) where Emin denotes the elastic modulus of the Ersatz material, an approximation for void material to prevent
singularity of the stiffness matrix, 0 < ρe < 1 , limits corresponding to Ersatz and solid materials, p is a penaliza-
tion factor for driving the density towards the black and white solution and E0 denotes the elastic modulus of
solid material.h The deterministic optimization problem can be expressed as The deterministic optimization problem can be expressed as (9)
minimize
ρ
c(EH
ijkl(ρ))
subject to KuA(kl) = f kl, k, l = 1, . . . , d
N
e=1
veρe/|Y| ≤v, 0 ≤ρe ≤1, e = 1, . . . , N. (9) where the objective function c(EH
ijkl) is a function of the homogenized stiffness tensors, K is the global stiffness
matrix, uA(kl) and f kl are the global displacement and external force vectors of the test case (kl), respectively,
d, ve and v denote the spatial dimension, element volume and upper limit of the volume fraction, respectively. Methodology
Th
k gy
The work integrates the material uncertainty model (simulated by in-house developed codes) with deterministic
topology optimization36 to develop a parallel Monte Carlo based robust topology optimization framework. A
flow-diagram of the proposed RTO framework is presented in Fig. 2.The main intent is to illustrate the effect of
material uncertainties on the micro-structural topologies and derive robust configurations for extreme material
design. As shown in Fig. 2, the entire methodology comprises of three steps: (1) the user-defined input module
where the system and the optimization parameters are initialized, (2) the analysis module where the expensive
computations take place involving uncertainty quantification nested within the optimization loop. To reduce the
computational cost, a parallel version of Monte Carlo simulation is employed to simulate the randomness within
each optimization iteration. The details of this step regarding the optimization algorithm, finite element model,
energy-based homogenization and sensitivity computations are presented subsequently. (3) The post-processing
module is where the resulting robust micro-structural topologies with extreme properties are exported to a CAD
software for 3D printing. Homogenization. Considering linear elastic behaviour, the equivalent constitutive behaviour of periodic
structures can be determined by homogenization technique. This work employs the energy-based homogeniza-
tion approach to predict the macroscopic equivalent elastic properties of micro-structures36. The approach is
based upon energy conservation with respect to stress and strain. The homogenized stiffness tensor EH
ijkl can be
expressed in terms of mutual energies as (1)
EH
ijkl =
1
|Y|
Y
EpqrsεA(ij)
pq
εA(kl)
rs
dY (1) where Y represents the base cell, Epqrs is the elasticity tensor in index notation, εA(ij)
pq and εA(kl)
rs
denote the super-
imposed strain fields. When the base cell is discretized into N finite elements, Eq. (1) can be approximated as where Y represents the base cell, Epqrs is the elasticity tensor in index notation, εA(ij)
pq and εA(kl)
rs
denote the super-
imposed strain fields. When the base cell is discretized into N finite elements, Eq. (1) can be approximated as https://doi.org/10.1038/s41598-021-94520-x https://doi.org/10.1038/s41598-021-94520-x https://doi.org/10.1038/s41598-021-94520-x Scientific Reports | (2021) 11:15221 | www.nature.com/scientificreports/ (2)
EH
ijkl =
1
|Y|
N
e=1
(uA(ij)
e
)TkeuA(kl)
e (2) where uA(kl)
e
are the element displacement solutions corresponding to the unit test strain fields ε0(kl) and ke
denotes the element stiffness matrix. It is to be noted that for 2-D problems, 11 ≡1 , 22 ≡2 , and 12 ≡3 . Thus,
Eq. Methodology
Th
k (2) can be concisely represented as where uA(kl)
e
are the element displacement solutions corresponding to the unit test strain fields ε0(kl) and ke
denotes the element stiffness matrix. It is to be noted that for 2-D problems, 11 ≡1 , 22 ≡2 , and 12 ≡3 . Thus,
Eq. (2) can be concisely represented as (3)
Qij =
1
|Y|
N
e=1
q(ij)
e (3)
Qij =
1
|Y|
N
e=1
q(ij)
e (3) where Qij is given by where Qij is given by where Qij is given by
Q
Q
Q
EH
EH
EH
where Qij is given by
(4)
Q11 Q12 Q13
Q21 Q22 Q23
=
EH
1111 EH
1122 EH
1112
EH
EH
EH
(4)
Q11 Q12 Q13
Q21 Q22 Q23
Q31 Q32 Q33
=
EH
1111 EH
1122 EH
1112
EH
2211 EH
2222 EH
2212
EH
1211 EH
1222 EH
1212
(4)
Q11 Q12 Q13
Q21 Q22 Q23
Q31 Q32 Q33
=
EH
1111 EH
1122 EH
1112
EH
2211 EH
2222 EH
2212
EH
1211 EH
1222 EH
1212
(4) In Eq. (3), q(ij)
e are the element mutual energies and can be expressed as In Eq. (3), q(ij)
e are the element mutual energies and can be expressed as (5)
q(ij)
e
= (uA(i)
e
)TkeuA(j)
e (5) Periodic boundary conditions.
The displacements on a pair of opposite boundaries of a 2-D base cell The motive of this work is to determine the extreme properties of material micro-structures, such as maxi-
mum bulk modulus, maximum shear modulus, minimum Poisson’s ratio and maximum equivalent elastic modu-
lus The corresponding objecti e functions can be defined as where the objective function c(EH
ijkl) is a function of the homogenized stiffness tensors, K is the global stiffness
matrix, uA(kl) and f kl are the global displacement and external force vectors of the test case (kl), respectively,
d, ve and v denote the spatial dimension, element volume and upper limit of the volume fraction, respectively. The motive of this work is to determine the extreme properties of material micro-structures, such as maxi- where the objective function c(EH
ijkl) is a function of the homogenized stiffness tensors, K is the global stiffness
matrix, uA(kl) and f kl are the global displacement and external force vectors of the test case (kl), respectively,
d, ve and v denote the spatial dimension, element volume and upper limit of the volume fraction, respectively.h y
The motive of this work is to determine the extreme properties of material micro-structures, such as maxi-
mum bulk modulus, maximum shear modulus, minimum Poisson’s ratio and maximum equivalent elastic modu-
lus. The corresponding objective functions can be defined as • Design of maximum bulk modulus: (10) c = −(E1111 + E1122 + E2211 + E2222) (10)
c = −(E1111 + E1122 + E2211 + E2222) • Design of maximum shear modulus: Scientific Reports | (2021) 11:15221 | https://doi.org/10.1038/s41598-021-94520-x www.nature.com/scientificreports/ c = −E1212 (11) • Design of minimum Poisson’s ratio: • Design of minimum Poisson’s ratio: ent elastic modulus:
(12)
c = E1122 + E2211 −δE1212 (12
c = E1122 + E2211 −δE1212 c = E1122 + E2211 −δE1212 (12) • Design of maximum equivalent elastic modulus: • Design of maximum equivalent elastic modulus: (13)
c = −1
2(E1111 + E2222) c = −1
2(E1111 + E2222) (13) Note that the topological design of minimum and negative Poisson’s ratio (E1122/E1111) has always been a
difficult issue. It is observed from the literature that auxetic properties can be obtained by imposing additional
constraints on isotropy or bulk modulus36. An equivalent expression in the form of Eq. (12) is used for this pur-
pose and adopted from23. Selection of the term δ in Eq. Periodic boundary conditions.
The displacements on a pair of opposite boundaries of a 2-D base cell (12) is discussed later in the numerical illustration section.h p
p
q
The solution scheme for solving the equilibrium equation in Eq. (9) adopted here can be found in Xia and
Breitkopf36. After obtaining the finite element displacement solution, the optimization problem in Eq. (9) is
solved by using optimality criteria method. The heuristic updating criterion13 is expressed as (14)
ρnew
e
= max(0, ρe −m), if ρeBη
e ≤max(0, ρe −m)
min(0, ρe + m), if ρeBη
e ≥min(0, ρe + m)
ρeBη
e , otherwise (14) where m is the step size, η is the damping coefficient and Be can be determined by using the following optimality
condition where m is the step size, η is the damping coefficient and Be can be determined by using the following optimality
ondition (15)
Be =
−∂c
∂ρe
∂V
∂ρe (15) where is the Lagrange multiplier. It imposes the satisfaction of the material volume fraction constraint. The
numerator ∂c
∂ρe which is the sensitivity of the objective function can be determined by using the adjoint method as where is the Lagrange multiplier. It imposes the satisfaction of the material volume fraction constraint. The
numerator ∂c
∂ρe which is the sensitivity of the objective function can be determined by using the adjoint method as (16)
∂EH
ijkl
∂ρe
=
1
|Y|pρp−1
e
(E0 −Emin)(uA(ij)
e
)Tk0uA(kl)
e (16) where k0 represent the element stiffness matrix with unit elastic modulus. For a uniform mesh, the element vol-
ume ve = 1 and hence, ∂V
∂ρe = 1 . To ensure convergence of the optimization problem Eq. (9), filtering techniques
are used which are illustrated next. where k0 represent the element stiffness matrix with unit elastic modulus. For a uniform mesh, the element vol-
ume ve = 1 and hence, ∂V
∂ρe = 1 . To ensure convergence of the optimization problem Eq. (9), filtering techniques
are used which are illustrated next. Filtering. To eliminate the formation of a checkerboard pattern and resolve the mesh-dependent issue, a
filter is applied either to the sensitivities or the densities38,39. The sensitivity based filtering modifies the sensitivi-
ties ∂c
∂ρe (Eq. (16) as, (17)
∂c
∂ρe
=
1
max(γ , ρe)
i∈Ne Hei
i∈Ne
Heiρi
∂c
∂ρi (17) where γ = 10−3 is a small positive number incorporated to avoid the denominator becoming zero. Hei is a
weight term defined in Eq. (18). Sensitivity calculation.
Differentiating cRTO , we obtain (28), it is obvious that the topological derivatives for RTO can be computed based on conventional/
deterministic topological derivatives. From Eq. (28), it is obvious that the topological derivatives for RTO can be computed based on conventio
deterministic topological derivatives. Sensitivity calculation.
Differentiating cRTO , we obtain Sensitivity calculation. Differentiating cRTO , we obtain Sensitivity calculation. Differentiating cRTO , we obtain (22)
∂cRTO
∂de
= ∂µc
∂de
+ 3∂σc
∂de
. (22) Now considering the first term of Eq. (22) and expanding it, we obtain Now considering the first term of Eq. (22) and expanding it, we obtain (23)
∂µc
∂de
=
∂
∂de
1
Nsimu
Nsimu
i=1
ci
=
1
Nsimu
Nsimu
i=1
∂ci
∂de
= E
∂c
∂de
(23) where we have represented the expectation based on Nsimu number of samples. From Eq. (23), we can see that the
first term of Eq. (22) can be computed based on the expectation of the topological derivatives in the deterministic
case. For the second term in Eq. (22), we have where we have represented the expectation based on Nsimu number of samples. From Eq. (23), we can see that the
first term of Eq. (22) can be computed based on the expectation of the topological derivatives in the deterministic
case. For the second term in Eq. (22), we have (24)
∂σc
∂de
= ∂√var(c)
∂de
=
1
2√var(c)
∂
∂de
[var (c)] =
1
2√var(c)
∂
∂de
E
c2
−E(c)2
=
1
2√var(c)
∂
∂de
E
c2
−
∂
∂de
E(c)2
(24) where var (•) indicates the variance operator. Considering the first term in Eq. (24) and expanding it similar to
Eq. (23), we have (25)
∂
∂de
E
c2
= ∂
∂de
1
Nsimu
Nsimu
i=1
c2
i
=
1
Nsimu
Nsimu
i=1
∂c2
i
∂de
=
1
Nsimu
Nsimu
i=1
2ci
∂ci
∂de
= 2E
c ∂c
∂de
. (25) Finally, considering the second term in Eq. (24) and using Eq. (23), we have (26)
∂
∂de
E(c)2 = 2E(c) ∂
∂de
E(c) = 2E(c)E
∂c
∂de
. (26) Substituting Eqs. (26) and (25) into Eq. (24), we obtain Substituting Eqs. (26) and (25) into Eq. (24), we obtain (27)
∂σc
∂de
=
1
√var(c)
E
c ∂c
∂de
−E(c)E
∂c
∂de
. (27) Finally, substituting Eqs. (27) and (23) into Eq. (22), we obtain Finally, substituting Eqs. (27) and (23) into Eq. (22), we obtain (28)
∂cRTO
∂de
= E
∂c
∂de
+ 3
1
√var(c)
E
c ∂c
∂de
−E(c)E
∂c
∂de
. (28) From Eq. Periodic boundary conditions.
The displacements on a pair of opposite boundaries of a 2-D base cell Ne represents the set of elements i whose centre-to-centre distance to element
e, (e, i) , is less than the filter radius rmin . The size of the filter radius can be used to control the minimum size
of the emerging features in the design domain. Hei = max(0, rmin − (e, i)) (18)
Hei = max(0, rmin − (e, i)) (18) The density based filtering converts the original densities to the filtered densities ρe as, The density based filtering converts the original densities to the filtered densities ρe as, (19)
˜ρe =
1
i∈Ne Hei
i∈Ne
Heiρi (19) The sensitivities with respect to ρj can be obtained as, The sensitivities with respect to ρj can be obtained as, (20)
∂c
∂ρj
=
e∈Nj
∂c
∂˜ρe
∂˜ρe
∂ρj
=
e∈Nj
1
i∈Ne Hei
Hej
∂c
∂˜ρe (20) Note that ∂c
∂˜ρe can be evaluated by Eq. (16) upon replacing ρe with ˜ρe. Note that ∂c
∂˜ρe can be evaluated by Eq. (16) upon replacing ρe with ˜ρe. Note that ∂c
∂˜ρe can be evaluated by Eq. (16) upon replacing ρe with ˜ρe. Robust topology optimization. The objective function considered for the robust topology optimization
s Robust topology optimization. The objective function considered for the robust topology optimization
is https://doi.org/10.1038/s41598-021-94520-x https://doi.org/10.1038/s41598-021-94520-x https://doi.org/10.1038/s41598-021-94520-x Scientific Reports | (2021) 11:15221 | www.nature.com/scientificreports/ (21)
cRTO = µc + 3σc (21
cRTO = µc + 3σc cRTO = µc + 3σc (21) where µc and σc are the mean and standard deviation of the deterministic objective function c, respectively. Numerical illustration: results and discussion
l
i
d
il They include, (1) deterministic design, (2) robust design considering parametric uncertainty models for elastic
modulus and Poisson’s ratio, (3) robust design considering random field model for elastic modulus and (4)
robust design considering a combination of random field and parametric uncertainty models for the elastic
modulus and Poisson’s ratio, respectively. Note that the terms of the equivalent elastic matrix (given by Eq. (4))
represent their mean values. The standard deviation of the matrix terms is consistent with the level of input
uncertainty and is observed to be within 5% bounds. The objective functions of the deterministic and robust
optimization have been evaluated by Eqs. (10) and (21), respectively. every optimization iteration to simulate the material variability. This has been carried out in a cost-effective
manner using parallel computing (employing all 8 cores of an Intel Xeon processor). Four case studies have
been undertaken to obtain extreme properties of material micro-structures, such as maximum bulk modulus,
maximum shear modulus, minimum negative Poisson’s ratio (auxetic metamaterials) and maximum equivalent
elastic modulus. every optimization iteration to simulate the material variability. This has been carried out in a cost-effective
manner using parallel computing (employing all 8 cores of an Intel Xeon processor). Four case studies have
been undertaken to obtain extreme properties of material micro-structures, such as maximum bulk modulus,
maximum shear modulus, minimum negative Poisson’s ratio (auxetic metamaterials) and maximum equivalent
elastic modulus. The initial design is the same for all the following cases studied (except type III). The nominal values of the
elastic modulus and Poisson’s ratio at the element level are adopted as 1 and 0.3, respectively. For the stochastic
cases, their mean values are same as that of the nominal values. The length scale used for discretizing the random
field model is taken as 0.2. The results are represented by reporting the optimal unit cell, 3 × 4 periodic arrange-
ment, equivalent elastic matrices and robust optimal solutions for each of the extreme material configurations. Maximum bulk modulus (Type I). Table 1 illustrates the optimal solutions for the case of type I. To obtain
these results, density based filtering and the following parameter values are considered: p = 5 and, rmin = 2. It can be observed from Table 1 that uncertainty affects both the topological configuration and the equivalent
elastic matrix. Numerical illustration: results and discussion
l
i
d
il They include, (1) deterministic design, (2) robust design considering parametric uncertainty models for elastic
modulus and Poisson’s ratio, (3) robust design considering random field model for elastic modulus and (4)
robust design considering a combination of random field and parametric uncertainty models for the elastic
modulus and Poisson’s ratio, respectively. Note that the terms of the equivalent elastic matrix (given by Eq. (4))
represent their mean values. The standard deviation of the matrix terms is consistent with the level of input
uncertainty and is observed to be within 5% bounds. The objective functions of the deterministic and robust
optimization have been evaluated by Eqs. (10) and (21), respectively. www.nature.com/scientificreports/ Table 1. Optimal solutions for maximum bulk modulus (Type I). Results from four case studies are shown. They include, (1) deterministic design, (2) robust design considering parametric uncertainty models for elastic
modulus and Poisson’s ratio, (3) robust design considering random field model for elastic modulus and (4)
robust design considering a combination of random field and parametric uncertainty models for the elastic
modulus and Poisson’s ratio, respectively. Note that the terms of the equivalent elastic matrix (given by Eq. (4))
represent their mean values. The standard deviation of the matrix terms is consistent with the level of input
uncertainty and is observed to be within 5% bounds. The objective functions of the deterministic and robust
optimization have been evaluated by Eqs. (10) and (21), respectively. Table 1. Optimal solutions for maximum bulk modulus (Type I). Results from four case studies are shown. They include, (1) deterministic design, (2) robust design considering parametric uncertainty models for elastic
modulus and Poisson’s ratio, (3) robust design considering random field model for elastic modulus and (4)
robust design considering a combination of random field and parametric uncertainty models for the elastic
modulus and Poisson’s ratio, respectively. Note that the terms of the equivalent elastic matrix (given by Eq. (4))
represent their mean values. The standard deviation of the matrix terms is consistent with the level of input
uncertainty and is observed to be within 5% bounds. The objective functions of the deterministic and robust
optimization have been evaluated by Eqs. (10) and (21), respectively. Table 1. Optimal solutions for maximum bulk modulus (Type I). Results from four case studies are shown. Numerical illustration: results and discussion
l
i
d
il Implementation details. In this work, the design domain is considered to be square and discretized into
square plane stress elements. The volume fraction is taken to be 0.5. The unit cell is discretized into 3 × 3 ele-
ments. The influence of an initial design can affect the final optimal topology significantly. For this work, a
square shaped void region at the center of the unit cell is adopted as the initial design40 as shown in Fig. 2. The
small void is generally inserted in the initial design to stimulate the evolutionary optimization process. g
y
g
y p
p
It has been reported in multiple studies that topology optimization for the design of extreme material proper-
ties is prone to multiple local minima. The final optimal solution is found to be extremely sensitive to the initial
topology, shape of the unit cell, filter radius (rmin) , penalization factor (p), filtering approach (sensitivity filtering
or density filtering) and others. Therefore, for selecting the appropriate parameter values, few rules of thumb have
been followed. The optimization problem Eq. (9) is non-convex for p ≥1 . Although the high values of penaliza-
tion factor will result in more distinct topologies, the algorithm is likely to get trapped in a local minima17. A
smaller filter radius value generally results in a better solution (in terms of a detailed micro-structure) as higher
frequency details are visible through low-pass filter. Compared to sensitivity filtering, density filtering is observed
to yield less sensitive designs and hence preferable for material micro-structure design. y
g
p
g
Parametric study has been carried out by considering various combinations of stochastic material parameter
models such as, spatial variation via random field and random parametric models. The stochastic models assumed
are in line with the literature (discussed in the introduction section) where spatially varying and parametric
models simulate non-homogenous and uniform manufacturing errors, respectively. For stochastic simulations,
the elastic modulus and Poisson’s ratio are considered to be random and follow log-normal distribution with 5 %
variability. It is practical to assume that all realizations corresponding to these random parameters will be posi-
tive and therefore, they are assumed to be log-normally distributed. In doing so, 1000 MCS is performed within Scientific Reports | (2021) 11:15221 | https://doi.org/10.1038/s41598-021-94520-x nature.com/scientificreports/
Table 1. Optimal solutions for maximum bulk modulus (Type I). Results from four case studies are shown. Numerical illustration: results and discussion
l
i
d
il They include, (1) deterministic design, (2) robust design considering parametric uncertainty models for elastic
modulus and Poisson’s ratio, (3) robust design considering random field model for elastic modulus and (4)
robust design considering a combination of random field and parametric uncertainty models for the elastic
modulus and Poisson’s ratio, respectively. Note that the terms of the equivalent elastic matrix (given by Eq. (4))
represent their mean values. The standard deviation of the matrix terms is consistent with the level of input
uncertainty and is observed to be within 5 % bounds. The objective functions of the deterministic and robust
optimization have been evaluated by Eqs. (11) and (21), respectively. dom field models, the difference is significant for the hybrid uncertainty model, exhibiting strong anisotropic
behaviour. In terms of mean value of the objective function, the random field cases lead to higher value of bulk
modulus compared to the deterministic and parametric uncertainty cases. Also, the random field cases result
in a more robust topological design indicated by lower values of the standard deviation of objective function. dom field models, the difference is significant for the hybrid uncertainty model, exhibiting strong anisotropic
behaviour. In terms of mean value of the objective function, the random field cases lead to higher value of bulk
modulus compared to the deterministic and parametric uncertainty cases. Also, the random field cases result
in a more robust topological design indicated by lower values of the standard deviation of objective function. Maximum shear modulus (Type II). Table 2 illustrates the optimal solutions for the case of type II. To obtain these results, density based filtering and the following parameter values are considered: p = 5 and,
rmin = 2.It can be observed from Table 2 that uncertainty affects both the topological configuration and the
equivalent elastic matrix. The stiffness terms (diagonal entries) Q11 , Q22 and Q33 and the off-diagonal term Q12 are
observed to increase for the random field and hybrid input uncertainty models compared to the deterministic
case. These terms achieve lower values for the parametric uncertainty model compared to the deterministic case. Interestingly, for the random field and hybrid input uncertainty models, the resulting structure show (1) non-
zero values of Q13 and Q23 terms and (2) Q11 = Q22 . (1) indicates coupling of axial and shear deformations/stress
states and (2) illustrates anisotropic behaviour. Numerical illustration: results and discussion
l
i
d
il The stiffness terms Q11 and Q22 are observed to increase for all types of uncertainty models com-
pared to the deterministic case. Specifically, the greatest rise is observed for the random field cases. The term Q33
decreases due to uncertainty. The off-diagonal term Q12 reduces for all uncertainty models and the deterministic
model achieves the maximum value. Interestingly, for the hybrid input uncertainty modelling case, the resulting
structure shows non-zero (negative) Q13 and Q23 terms. This indicates coupling of axial and shear deformations/
stress states. Although there is a slight difference between the terms Q11 and Q22 for the deterministic and ran- https://doi.org/10.1038/s41598-021-94520-x Scientific Reports | (2021) 11:15221 | www.nature.com/scientificreports/ Table 2. Optimal solutions for maximum shear modulus (Type II). Results from four case studies are shown. They include, (1) deterministic design, (2) robust design considering parametric uncertainty models for elastic
modulus and Poisson’s ratio, (3) robust design considering random field model for elastic modulus and (4)
robust design considering a combination of random field and parametric uncertainty models for the elastic
modulus and Poisson’s ratio, respectively. Note that the terms of the equivalent elastic matrix (given by Eq. (4))
represent their mean values. The standard deviation of the matrix terms is consistent with the level of input
uncertainty and is observed to be within 5 % bounds. The objective functions of the deterministic and robust
optimization have been evaluated by Eqs. (11) and (21), respectively. Table 2. Optimal solutions for maximum shear modulus (Type II). Results from four case studies are shown. They include, (1) deterministic design, (2) robust design considering parametric uncertainty models for elastic
modulus and Poisson’s ratio, (3) robust design considering random field model for elastic modulus and (4)
robust design considering a combination of random field and parametric uncertainty models for the elastic
modulus and Poisson’s ratio, respectively. Note that the terms of the equivalent elastic matrix (given by Eq. (4))
represent their mean values. The standard deviation of the matrix terms is consistent with the level of input
uncertainty and is observed to be within 5 % bounds. The objective functions of the deterministic and robust
optimization have been evaluated by Eqs. (11) and (21), respectively. Table 2. Optimal solutions for maximum shear modulus (Type II). Results from four case studies are shown. Numerical illustration: results and discussion
l
i
d
il They include, (1) deterministic design (column 3), (2) robust design
considering parametric uncertainty models for elastic modulus and Poisson’s ratio (column 4), (3) robust
design considering random field model for elastic modulus (column 5) and (4) robust design considering a
hybrid combination of random field and parametric uncertainty models for the elastic modulus and Poisson’s
ratio (column 6). Note that the mean Poisson’s ratio is computed by (E1122/E1111) from the mean equivalent
elastic matrix. The objective functions of the deterministic and robust optimization have been evaluated by
Eqs. (12) and (21), respectively. Table 3. Parametric study to determine δ in Eq. (12) to achieve minimum negative Poisson’s ratio (Type III). Results from four case studies are shown. They include, (1) deterministic design (column 3), (2) robust design
considering parametric uncertainty models for elastic modulus and Poisson’s ratio (column 4), (3) robust
design considering random field model for elastic modulus (column 5) and (4) robust design considering a
hybrid combination of random field and parametric uncertainty models for the elastic modulus and Poisson’s
ratio (column 6). Note that the mean Poisson’s ratio is computed by (E1122/E1111) from the mean equivalent
elastic matrix. The objective functions of the deterministic and robust optimization have been evaluated by
Eqs. (12) and (21), respectively. Minimum Poisson’s ratio (Type III). Density based filtering and the following parameter values are con-
sidered: p = 5 and, rmin = 5 , for the type III case. The unit cell is discretized into 200 elements along both the
horizontal and vertical directions. As the term δ (refer Eq. (12)) causes significant sensitivity to the topological
design of auxetic metamaterials23, a parametric study is performed in Table 3 to investigate the effect of δ on
the resulting topologies with minimum Poisson’s ratio.It can be observed from Table 3 that out of the adopted
δ values, only δ = 0.5 and 1 lead to negative mean Poisson’s ratio corresponding to the deterministic and each
of the stochastic models. δ = 0.5 has been selected as lower Poisson’s ratios are obtained compared to the case δ
= 1. The corresponding row has been highlighted to indicate the best and the most consistent performance in
Table 3 for clarity. Numerical illustration: results and discussion
l
i
d
il Table 4 illustrates the optimal solutions for the case of type III, corresponding to δ = 0.5.It can
be observed from Table 4 that uncertainty affects both the topological configuration and the equivalent elastic
matrix. Although the overall resulting topologies from the deterministic and parametric uncertainty models
are very similar with fine differences at the boundary. The stiffness terms Q11 and Q22 are observed to increase
for the random field and hybrid uncertainty models and decrease for parametric uncertainty compared to the
deterministic case. The stiffness term Q33 is observed to increase for all stochastic models compared to the
deterministic case. The off-diagonal term Q12 is negative for all the cases reported and observed to increase for
the parametric stochastic model and decrease for the random field and hybrid uncertainty cases than that of the
deterministic one. Interestingly, for the random field and hybrid input uncertainty models: (1) the terms Q13
and Q23 are non-zero (negative), which illustrates coupling of axial and shear deformations/stress states and (2)
Q11 = Q22 , which indicates anisotropic behaviour. The random field and hybrid input uncertainty models lead
to lower values of the mean objective function compared to the deterministic and parametric uncertainty cases. Specifically, the parametric and hybrid stochastic models result in the maximum and minimum value of Pois-
son’s ratio, respectively. The deterministic, random field and hybrid input uncertainty models result in the same
level of robustness of the topological design indicated by the standard deviation of the objective function. The
parametric uncertainty model achieves the most robust design by a small margin. The results in Table 4 illustrate
that negative Poisson’s ratio (given by Q12/Q11 ) has been achieved for all the case studies undertaken. This is
worth mentioning as the design of auxetic metamaterials by topology optimization is a challenging task on its
own and even more difficult with uncertainties. Maximum equivalent elastic modulus (Type IV). Table 5 illustrates the optimal solutions for the case
of type IV. To obtain these results, density based filtering and the following parameter values are considered:
p = 5 and, rmin = 2.It can be observed from Table 5 that uncertainty affects both the topological configuration
and the equivalent elastic matrix. The resulting topologies obtained from the random field and hybrid random
input models are the same. Numerical illustration: results and discussion
l
i
d
il In terms of mean value of the objective function, the random field
and hybrid input uncertainty models lead to higher value of shear modulus compared to the deterministic and
parametric uncertainty cases. The parametric stochastic model results in the minimum value of shear modulus. Also, the random field and hybrid input uncertainty models result in a more robust topological design indicated
by lower values of the standard deviation of objective function. https://doi.org/10.1038/s41598-021-94520-x Scientific Reports | (2021) 11:15221 | www.nature.com/scientificreports/ Table 3. Parametric study to determine δ in Eq. (12) to achieve minimum negative Poisson’s ratio (Type III). Results from four case studies are shown. They include, (1) deterministic design (column 3), (2) robust design
considering parametric uncertainty models for elastic modulus and Poisson’s ratio (column 4), (3) robust
design considering random field model for elastic modulus (column 5) and (4) robust design considering a
hybrid combination of random field and parametric uncertainty models for the elastic modulus and Poisson’s
ratio (column 6). Note that the mean Poisson’s ratio is computed by (E1122/E1111) from the mean equivalent
elastic matrix. The objective functions of the deterministic and robust optimization have been evaluated by
Eqs. (12) and (21), respectively. Numerical illustration: results and discussion
l
i
d
il δ
Response statistics
Deterministic
Robust (parametric
uncertainty)
Robust (random field)
Robust (hybrid
uncertainty)
0.5
Mean of objective function
− 0.0392
− 0.0336
− 0.0950
− 0.1024
SD of objective function
0.0019
0.0017
0.0019
0.0019
Mean of Poisson’s ratio
− 0.4915
− 0.4791
− 0.5895
− 0.5908
1.0
Mean of objective function
− 0.0405
− 0.0355
− 0.1098
− 0.1110
SD of objective function
0.0020
0.0018
0.0025
0.0025
Mean of Poisson’s ratio
− 0.4168
− 0.3293
− 0.5468
− 0.5518
1.5
Mean of objective function
− 0.0559
− 0.0508
− 0.1260
− 0.1249
SD of objective function
0.0028
0.0025
0.0028
0.0029
Mean of Poisson’s ratio
0.0965
0.0831
− 0.6120
− 0.5376
2.0
Mean of objective function
− 0.0912
− 0.0828
− 0.1304
− 0.1336
SD of objective function
0.0045
0.0041
0.0030
0.0031
Mean of Poisson’s ratio
0.1423
0.1553
− 0.5269
− 0.5318
2.5
Mean of objective function
− 0.1190
− 0.1142
− 0.1274
− 0.1260
SD of objective function
0.0059
0.0065
0.0029
0.0029
Mean of Poisson’s ratio
0.1552
0.1769
− 0.3299
− 0.3872 δ
Response statistics
Deterministic
Robust (parametric
uncertainty)
Robust (random field)
Robust (hybrid
uncertainty)
0.5
Mean of objective function
− 0.0392
− 0.0336
− 0.0950
− 0.1024
SD of objective function
0.0019
0.0017
0.0019
0.0019
Mean of Poisson’s ratio
− 0.4915
− 0.4791
− 0.5895
− 0.5908
1.0
Mean of objective function
− 0.0405
− 0.0355
− 0.1098
− 0.1110
SD of objective function
0.0020
0.0018
0.0025
0.0025
Mean of Poisson’s ratio
− 0.4168
− 0.3293
− 0.5468
− 0.5518
1.5
Mean of objective function
− 0.0559
− 0.0508
− 0.1260
− 0.1249
SD of objective function
0.0028
0.0025
0.0028
0.0029
Mean of Poisson’s ratio
0.0965
0.0831
− 0.6120
− 0.5376
2.0
Mean of objective function
− 0.0912
− 0.0828
− 0.1304
− 0.1336
SD of objective function
0.0045
0.0041
0.0030
0.0031
Mean of Poisson’s ratio
0.1423
0.1553
− 0.5269
− 0.5318
2.5
Mean of objective function
− 0.1190
− 0.1142
− 0.1274
− 0.1260
SD of objective function
0.0059
0.0065
0.0029
0.0029
Mean of Poisson’s ratio
0.1552
0.1769
− 0.3299
− 0.3872 Table 3. Parametric study to determine δ in Eq. (12) to achieve minimum negative Poisson’s ratio (Type III). Results from four case studies are shown. Table 3. Parametric study to determine δ in Eq. (12) to achieve minimum negative Poisson’s ratio (Type III).
Results from four case studies are shown. They include, (1) deterministic design (column 3), (2) robust design
considering parametric uncertainty models for elastic modulus and Poisson’s ratio (column 4), (3) robust
design considering random field model for elastic modulus (column 5) and (4) robust design considering a
hybrid combination of random field and parametric uncertainty models for the elastic modulus and Poisson’s
ratio (column 6). Note that the mean Poisson’s ratio is computed by (E1122/E1111) from the mean equivalent
elastic matrix. The objective functions of the deterministic and robust optimization have been evaluated by
Eqs. (12) and (21), respectively. Numerical illustration: results and discussion
l
i
d
il They include, (1) deterministic design, (2) robust design considering parametric uncertainty models for elastic
modulus and Poisson’s ratio, (3) robust design considering random field model for elastic modulus and (4)
robust design considering a combination of random field and parametric uncertainty models for the elastic
modulus and Poisson’s ratio. Note that the terms of the equivalent elastic matrix (given by Eq. (4)) represent
their mean values. The standard deviation of the matrix terms is consistent with the level of input uncertainty
and is observed to be within 5 % bounds. The objective functions of the deterministic and robust optimization
have been evaluated by Eqs. (12) and (21), respectively. Table 4. Optimal solutions for minimum Poisson’s ratio (Type III). Results from four case studies are shown. They include, (1) deterministic design, (2) robust design considering parametric uncertainty models for elastic
modulus and Poisson’s ratio, (3) robust design considering random field model for elastic modulus and (4)
robust design considering a combination of random field and parametric uncertainty models for the elastic
modulus and Poisson’s ratio. Note that the terms of the equivalent elastic matrix (given by Eq. (4)) represent
their mean values. The standard deviation of the matrix terms is consistent with the level of input uncertainty
and is observed to be within 5 % bounds. The objective functions of the deterministic and robust optimization
have been evaluated by Eqs. (12) and (21), respectively. Table 4. Optimal solutions for minimum Poisson’s ratio (Type III). Results from four case studies are shown. They include, (1) deterministic design, (2) robust design considering parametric uncertainty models for elastic
modulus and Poisson’s ratio, (3) robust design considering random field model for elastic modulus and (4)
robust design considering a combination of random field and parametric uncertainty models for the elastic
modulus and Poisson’s ratio. Note that the terms of the equivalent elastic matrix (given by Eq. (4)) represent
their mean values. The standard deviation of the matrix terms is consistent with the level of input uncertainty
and is observed to be within 5 % bounds. The objective functions of the deterministic and robust optimization
have been evaluated by Eqs. (12) and (21), respectively. uncertainty models, the resulting structure shows anisotropic property indicated by Q11 = Q22 and illustrates
coupling of axial and shear deformations/stress states indicated by non-zero Q13 term. Scientific Reports | (2021) 11:15221 | Numerical illustration: results and discussion
l
i
d
il The stiffness terms (diagonal entries) Q11 , Q22 and Q33 are observed to increase for
the random field compared to the deterministic case. The stiffness terms Q11 and Q33 are observed to increase
for the hybrid random input model compared to the deterministic case. These terms achieve lower values for
the parametric uncertainty model compared to the deterministic case. The off-diagonal term Q12 is observed
to be higher for all the stochastic models than that of the deterministic one. Interestingly, for the hybrid input https://doi.org/10.1038/s41598-021-94520-x Scientific Reports | (2021) 11:15221 | www.nature.com/scientificreports/ Table 4. Optimal solutions for minimum Poisson’s ratio (Type III). Results from four case studies are shown. They include, (1) deterministic design, (2) robust design considering parametric uncertainty models for elastic
modulus and Poisson’s ratio, (3) robust design considering random field model for elastic modulus and (4)
robust design considering a combination of random field and parametric uncertainty models for the elastic
modulus and Poisson’s ratio. Note that the terms of the equivalent elastic matrix (given by Eq. (4)) represent
their mean values. The standard deviation of the matrix terms is consistent with the level of input uncertainty
and is observed to be within 5 % bounds. The objective functions of the deterministic and robust optimization
have been evaluated by Eqs. (12) and (21), respectively. Table 4. Optimal solutions for minimum Poisson’s ratio (Type III). Results from four case studies are shown. They include, (1) deterministic design, (2) robust design considering parametric uncertainty models for elastic
modulus and Poisson’s ratio, (3) robust design considering random field model for elastic modulus and (4)
robust design considering a combination of random field and parametric uncertainty models for the elastic
modulus and Poisson’s ratio. Note that the terms of the equivalent elastic matrix (given by Eq. (4)) represent
their mean values. The standard deviation of the matrix terms is consistent with the level of input uncertainty
and is observed to be within 5 % bounds. The objective functions of the deterministic and robust optimization
have been evaluated by Eqs. (12) and (21), respectively. Table 4. Optimal solutions for minimum Poisson’s ratio (Type III). Results from four case studies are shown. Table 5. Optimal solutions for maximum equivalent elastic modulus (Type IV). Results from four case studies
are shown. They include, (1) deterministic design, (2) robust design considering parametric uncertainty models
for elastic modulus and Poisson’s ratio, (3) robust design considering random field model for elastic modulus
and (4) robust design considering a combination of random field and parametric uncertainty models for the
elastic modulus and Poisson’s ratio, respectively. Note that the terms of the equivalent elastic matrix (given by
Eq. (4)) represent their mean values. The standard deviation of the matrix terms is consistent with the level of
input uncertainty and is observed to be within 5 % bounds. The objective functions of the deterministic and
robust optimization have been evaluated by Eqs. (13) and (21), respectively. of the degree of sensitivity of input stochastic models on the optimal micro-structural topology. Thus, these
observations have a practical relevance as one needs to be more cautious in fabricating the metastructures
which follow a more sensitive input stochastic model.h p
The robust design leads to improved material properties compared to the deterministic design. l
l
l
l
h
b
f
d f
d
h
b
l d h
• Multiple interesting topological patterns have been found for guiding the robust metamaterial design. Impor-
tantly, all of the resulting topologies have tessellating property, which make them feasible for 3D printing. Thus, there can be a possible extension of the present computational work as the resulting robust topological
configuration of materials with extreme properties are practically realizable.i • Multiple interesting topological patterns have been found for guiding the robust metamaterial design. Impor-
tantly, all of the resulting topologies have tessellating property, which make them feasible for 3D printing. Thus, there can be a possible extension of the present computational work as the resulting robust topological
configuration of materials with extreme properties are practically realizable.i i
• In addition to the few resulting micro-structural configurations showing isotropic properties, interestingly,
the others yielded configurations which showed anisotropic property. Some cases led to positive and nega-
tive coupling of axial and shear deformations/stress states. These phenomenon were primarily observed for
the random field and hybrid uncertainty cases. This observation may be attributed to the fact that the spatial
variability (inhomogeneity) in the micro-structure induces anisotropy and coupling phenomena in the mate-
rial. Scientific Reports | (2021) 11:15221 | Numerical illustration: results and discussion
l
i
d
il They include, (1) deterministic design, (2) robust design considering parametric uncertainty models
for elastic modulus and Poisson’s ratio, (3) robust design considering random field model for elastic modulus
and (4) robust design considering a combination of random field and parametric uncertainty models for the
elastic modulus and Poisson’s ratio, respectively. Note that the terms of the equivalent elastic matrix (given by
Eq. (4)) represent their mean values. The standard deviation of the matrix terms is consistent with the level of
input uncertainty and is observed to be within 5 % bounds. The objective functions of the deterministic and
robust optimization have been evaluated by Eqs. (13) and (21), respectively. Table 5. Optimal solutions for maximum equivalent elastic modulus (Type IV). Results from four case studies
are shown. They include, (1) deterministic design, (2) robust design considering parametric uncertainty models
for elastic modulus and Poisson’s ratio, (3) robust design considering random field model for elastic modulus
and (4) robust design considering a combination of random field and parametric uncertainty models for the
elastic modulus and Poisson’s ratio, respectively. Note that the terms of the equivalent elastic matrix (given by
Eq. (4)) represent their mean values. The standard deviation of the matrix terms is consistent with the level of
input uncertainty and is observed to be within 5 % bounds. The objective functions of the deterministic and
robust optimization have been evaluated by Eqs. (13) and (21), respectively. Numerical illustration: results and discussion
l
i
d
il In terms of mean value of
the objective function, the random field and hybrid input uncertainty models lead to higher value of equivalent
elastic modulus compared to the deterministic and parametric uncertainty cases. The parametric stochastic
model results in the minimum value of equivalent elastic modulus. Also, the random field and hybrid input
uncertainty models result in a more robust topological design indicated by lower values of the standard devia-
tion of objective function. Conclusions. This section summarizes the results obtained from the numerical investigation. Few crit
points have been enumerated below: • It has been observed that the material uncertainties affect the resulting micro-structural topologies and
mechanical performance significantly.i • It has been observed that the material uncertainties affect the resulting micro-structural topologies and
mechanical performance significantly.i mechanical performance significantly. gi
y
• Out of the input stochastic material models, the random field model followed by the hybrid one lead to desir-
able performance in terms of (1) optimal mean performance, (2) most robust performance and (3) elastic
property, indicated by the mean value of objective function, standard deviation of objective function and
stiffness terms of the equivalent elastic matrix, respectively, for types I, II and IV.i i
• Out of the input stochastic material models, the random field model followed by the hybrid one lead to desir-
able performance in terms of (1) optimal mean performance, (2) most robust performance and (3) elastic
property, indicated by the mean value of objective function, standard deviation of objective function and
stiffness terms of the equivalent elastic matrix, respectively, for types I, II and IV.i f
q
,
p
y,
yp
,
• For type III, the hybrid uncertainty model followed by the random field leads to desirable output perfor-
mance (based upon combination of the above points (1)–(3)). This point along with the above are indicative f
q
p
y
yp
• For type III, the hybrid uncertainty model followed by the random field leads to desirable output perfor-
mance (based upon combination of the above points (1)–(3)). This point along with the above are indicative https://doi.org/10.1038/s41598-021-94520-x Scientific Reports | (2021) 11:15221 | www.nature.com/scientificreports/ Table 5. Optimal solutions for maximum equivalent elastic modulus (Type IV). Results from four case studies
are shown. It is also recommended to thoroughly scrutinize the existing equivalent expressions
for minimizing the Poisson’s ratio and is highly sensitive to the problem type or application in hand.hf • The effect of uncertainties on the material micro-structures is so high that even considering the accuracy and
high precision of additive manufacturing techniques, accounting nominal amount of uncertainty for mate-
rial design is recommended, at least in the conceptual design stage before the manufacturing. In fact, there
is nothing at stake, as it is shown in this study that robustness improves the design in terms of mechanical
performance. • The effect of uncertainties on the material micro-structures is so high that even considering the accuracy and
high precision of additive manufacturing techniques, accounting nominal amount of uncertainty for mate-
rial design is recommended, at least in the conceptual design stage before the manufacturing. In fact, there
is nothing at stake, as it is shown in this study that robustness improves the design in terms of mechanical
performance. Data availability
All
l
d y
All relevant data are available from the corresponding author upon reasonable request, and/or are included
within the main part and Supplementary Information. Received: 5 March 2021; Accepted: 6 July 2021 References
l 1. Solymar, L. & Shamonina, E. Waves in Metamaterials (Oxford University Press Inc, USA, 2009).t 2. Christensen, J., Kadic, M., Kraft, O. & Wegener, M. Vibrant times for mechanical metamaterials. M 2. Christensen, J., Kadic, M., Kraft, O. & Wegener, M. Vibrant times for mechanical metamaterials. Mrs Commun. 5, 453–462 (2015). 3. Hussein, M., Leamy, M. & Ruzzene, M. Dynamics of phononic materials and structures: Historical origins, recent progress, and
future outlook. Appl. Mech. Rev. 66, 040802 (2014). t
g
3. Hussein, M., Leamy, M. & Ruzzene, M. Dynamics of phononic materials and structures: Historical origins, recent progress, and
future outlook. Appl. Mech. Rev. 66, 040802 (2014). pp
4. Surjadi, J. U. et al. Mechanical metamaterials and their engineering applications. Adv. Eng. Mater. 21, 1800864 (2019). 5 L
Y Y
Y G
J K & S i
A 3 d h
i
l
i h l
id b
d
S i R
7 43407 (2017) 5. Lu, Y., Yang, Y., Guest, J. K. & Srivastava, A. 3-d phononic crystals with ultra-wide band gaps. Sci. Rep. 7, 43407 (2017).f 6. Yu, X., Zhou, J., Liang, H., Jiang, Z. & Wu, L. Mechanical metamaterials associated with stiffness, rigidity and compressibility: A
brief review. Prog. Mater. Sci. 94, 114–173 (2018). brief review. Prog. Mater. Sci. 94, 114–173 (2018). g
7. Choi, M.-J., Oh, M.-H., Koo, B. & Cho, S. Optimal design of lattice structures for controllable extremal band gaps. Sci. Rep. 9, 9976
(2019). 8. Mukhopadhyay, T., Naskar, S. & Adhikari, S. Anisotropy tailoring in geometrically isotropic multi-material lattices. Extreme Mech
Lett. 40, 100934 (2020). Lett. 40, 100934 (2020). 9. Karlicic, D., Cajic, M., Chatterjee, T. & Adhikari, S. Wave propagation in mass embedded and pre-stressed hexagonal lattices. Lett. 40, 100934 (2020). 9. Karlicic, D., Cajic, M., Chatterjee, T. & Adhikari, S. Wave propagation in mass embedded and pre-stressed hexagonal lattices. Compos Struct 256 113087 (2021) 9. Karlicic, D., Cajic, M., Chatterjee, T. & Adhikari, S. Wave propagation in mass embedded and pre-stressed hexagonal lattices
Compos. Struct. 256, 113087 (2021). p
0. Singh, A., Mukhopadhyay, T., Adhikari, S. & Bhattacharya, B. Equivalent in-plane voltage-dependent elastic moduli of piezoelectric
2d lattices. Int. J. Solids Struct. 208–209, 31–48 (2021). 1. Adhikari, S., Mukhopadhyay, T. & Liu, X. Broadband dynamic elastic moduli of honeycomb lattice materials: A generalized ana
lytical approach. Mech. Mater. 157, 103796 (2021). y
pp
12. Osanov, M. & Guest, J. K. References
l Topology optimization for architected materials design. Ann. Rev. Mater. Res. 46, 211–233 (201 12. Osanov, M. & Guest, J. K. Topology optimization fo 13. Bendsoe, M. P. & Sigmund, O. Topology Optimization y Optimization: Theory, Methods and Applications (Springer, Berlin g
p
gy
ph
y
pp
g
14. Xu, J., Gao, L., Xiao, M., Gao, J. & Li, H. Isogeometric topology optimization for rational design of ultra-lightweight archit
materials. Int. J. Mech. Sci. 166, 105103 (2020). J
,
(
)
5. Gibiansky, L. V. & Sigmund, O. Multiphase composites with extremal bulk modulus. J. Mech. Phys. Solids 48, 461–498 (2000). 15. Gibiansky, L. V. & Sigmund, O. Multiphase composites with e y
g
y
16. Huang, X., Radman, A. & Xie, Y. Topological design of microstructures of cellular materials for maximum bulk or shear modulus. Comput. Mater. Sci. 50, 1861–1870 (2011). p
7. Xia, L. Multiscale Structural Topology Optimization, 1st edn. (ISTE Press, Elsevier, 2016). https://doi.org/10.1016/C2015-0-01254-0 p
gy
p
p
g
18. Clausen, A., Wang, F., Jensen, J. S., Sigmund, O. & Lewis, J. A. Topology optimized architectures with programmable poisson’s
ratio over large deformations. Adv. Mater. 27, 5523–5527 (2015).f g
9. Long, K., Du, X., Xu, S. & Xie, Y. M. Maximizing the effective young’s modulus of a composite material by exploiting the poisson
effect. Compos. Struct. 153, 593–600 (2016). f
p
20. Zhang, H., Luo, Y. & Kang, Z. Bi-material microstructural design of chiral auxetic metamaterials using topology optimization. Compos. Struct. 195, 232–248 (2018). p
21. Chen, W. & Huang, X. Topological design of 3d chiral metamaterials based on couple-stress homogenization. J. Mech. Phys. Solids
131, 372–386 (2019). 2. Wang, Y., Liao, Z., Shi, S., Wang, Z. & Poh, L. H. Data-driven structural design optimization for petal-shaped auxetics using
isogeometric analysis. CMES Comput. Model. Eng. Sci. 122, 433–458 (2020). 3. Zheng, Y., Wang, Y., Lu, X., Liao, Z. & Qu, J. Evolutionary topology optimization for mechanical metamaterials with auxetic
property. Int. J. Mech. Sci. 179, 105638 (2020).f 24. Ye, M., Gao, L. & Li, H. A design framework for gradually stiffer mechanical metamaterial induced by negative poisson’s ratio
property. Mater. Des. 192, 108751 (2020). p
p
y
25. Sigmund, O. Manufacturing tolerant topology optimization. Acta Mechanica Sinica 25, 227–239 (2009). d
h d
d
b
f
l 26. Wang, F., Lazarov, B. S. & Sigmund, O. On projection methods, convergence and robust formulations in topology optimiza
Struct. Multidiscip. Although this requires further investigation and achieving this was unintentional but will prove to be
useful for designing material micro-structures with dual/simultaneous properties. i
• In addition to the few resulting micro-structural configurations showing isotropic properties, interestingly,
the others yielded configurations which showed anisotropic property. Some cases led to positive and nega-
tive coupling of axial and shear deformations/stress states. These phenomenon were primarily observed for
the random field and hybrid uncertainty cases. This observation may be attributed to the fact that the spatial
variability (inhomogeneity) in the micro-structure induces anisotropy and coupling phenomena in the mate-
rial. Although this requires further investigation and achieving this was unintentional but will prove to be
useful for designing material micro-structures with dual/simultaneous properties. g
g
p
p
• Considering the fact that the design of auxetic metamaterials by topology optimization is a challenging task
on its own and even becomes more difficult to obtain robust designs with micro-structural uncertainties, it
is worth noting that negative Poisson’s ratio was achieved for all the undertaken cases of type III using the
proposed framework by adopting appropriate parameters (Table 4). However, one should be careful in choos-
ing the parameters considering their high sensitivity on the resulting topologies (for example, δ of Eq. (12)) g
g
p
p
• Considering the fact that the design of auxetic metamaterials by topology optimization is a challenging task
on its own and even becomes more difficult to obtain robust designs with micro-structural uncertainties, it
is worth noting that negative Poisson’s ratio was achieved for all the undertaken cases of type III using the
proposed framework by adopting appropriate parameters (Table 4). However, one should be careful in choos-
ing the parameters considering their high sensitivity on the resulting topologies (for example, δ of Eq. (12)) https://doi.org/10.1038/s41598-021-94520-x Scientific Reports | (2021) 11:15221 | www.nature.com/scientificreports/ as evident from Table 3. It is also recommended to thoroughly scrutinize the existing equivalent expressions
for minimizing the Poisson’s ratio and is highly sensitive to the problem type or application in hand.hf as evident from Table 3. www.nature.com/scientificreports/ www.nature.com/scientificreports/ 35. Fabro, A. T., Meng, H. & Chronopoulos, D. Uncertainties in the attenuation performance of a multi-frequency metastructure from
additive manufacturing. Mech. Syst. Signal Process. 138, 106557 (2020). 5. Fabro, A. T., Meng, H. & Chronopoulos, D. Uncertainties in the attenuation performance of a multi-frequency metastructure from
additive manufacturing. Mech. Syst. Signal Process. 138, 106557 (2020). g
y
g
36. Xia, L. & Breitkopf, P. Design of materials using topology optimization and energy-based homogenization approach in ma
Struct. Multidiscip. Optim. 52, 1229–1241 (2015).i p
p
(
)
37. Xia, Z., Zhou, C., Yong, Q. & Wang, X. On selection of repeated unit cell model and application of unified periodic boundary
conditions in micro-mechanical analysis of composites. Int. J. Solids Struct. 43, 266–278 (2006).i p
p
(
)
7. Xia, Z., Zhou, C., Yong, Q. & Wang, X. On selection of repeated unit cell model and application of unified periodic boundary
conditions in micro-mechanical analysis of composites. Int. J. Solids Struct. 43, 266–278 (2006).i 8. Sigmund, O. Morphology-based black and white filters for topology optimization. Struct. Multidiscip. Optim. 33, 401–424 (2007)
9. Andreassen, E., Clausen, A., Schevenels, M., Lazarov, B. S. & Sigmund, O. Efficient topology optimization in matlab using 88 lines
of code. Struct. Multidiscip. Optim. 43, 1–16 (2011). 38. Sigmund, O. Morphology-based black and white filters for topology optimization. Struct. Multidiscip. Optim. 33, 401–424 (2007). 39. Andreassen, E., Clausen, A., Schevenels, M., Lazarov, B. S. & Sigmund, O. Efficient topology optimization in matlab using 88 lines
of code. Struct. Multidiscip. Optim. 43, 1–16 (2011). p
p
0. Amstutz, S., Giusti, S. M., Novotny, A. A. & de Souza Neto, E. A. Topological derivative for multi-scale linear elasticity models
applied to the synthesis of microstructures. Int. J. Numer. Methods Eng. 84, 733–756 (2010). p
p
40. Amstutz, S., Giusti, S. M., Novotny, A. A. & de Souza Neto, E. A. Topological derivative for multi-scale linear elasticity models
applied to the synthesis of microstructures. Int. J. Numer. Methods Eng. 84, 733–756 (2010). 40. Amstutz, S., Giusti, S. M., Novotny, A. A. & de Souza Neto, E. A. Topological derivative for multi-scale linear elasticity models
applied to the synthesis of microstructures. Int. J. Numer. Methods Eng. 84, 733–756 (2010). Acknowledgements g
Dr. T. Chatterjee and Prof. Michael I. Friswell gratefully acknowledge the support of the Engineering and Physi-
cal Sciences Research Council through the award of a Programme Grant “Digital Twins for Improved Dynamic
Design”, grant number EP/R006768. g
Dr. T. Chatterjee and Prof. Michael I. Friswell gratefully acknowledge the support of the Engineering and Physi-
cal Sciences Research Council through the award of a Programme Grant “Digital Twins for Improved Dynamic Dr. T. Chatterjee and Prof. Michael I. Friswell gratefully acknowledge the support of the Engineering and Physi-
cal Sciences Research Council through the award of a Programme Grant “Digital Twins for Improved Dynamic
”
b Competing interest h The authors declare no competing interests. The authors declare no competing interests. Author contributions TC conducted the experiments, generated the results and prepared the manuscript draft. All the authors reviewed
the manuscript, critically examined the results and checked the data and mathematical equations. References
l Optim. 43, 767–784 (2011). p
p
7. Schevenels, M., Lazarov, B. & Sigmund, O. Robust topology optimization accounting for spatially varying manufacturing errors
Comput. Methods Appl. Mech. Eng. 200, 3613–3627 (2011). p
pp
g
8. Jansen, M. et al. Robust topology optimization accounting for misplacement of material. Struct. Multidiscip. Optim. 47, 317–333
(2013). 9. Zheng, Y., Da, D., Li, H., Xiao, M. & Gao, L. Robust topology optimization for multi-material structures under interval uncertainty
Appl. Math. Model. 78, 627–647 (2020). pp
0. Wu, Y., Li, E., He, Z., Lin, X. & Jiang, H. Robust concurrent topology optimization of structure and its composite material consider-
ing uncertainty with imprecise probability. Comput. Methods Appl. Mech. Eng. 364, 112927 (2020). g
y
p
p
y
p
pp
g
31. Cai, J., Wang, C. & Fu, Z. Robust concurrent topology optimization of multiscale structure under single or multiple uncertain load
cases. Int. J. Numer. Methods Eng. 121, 1456–1483 (2020). cases. Int. J. Numer. Methods Eng. 121, 1456–1483 (2020). g
32. He, Z. C. et al. Robust topological design of actuator-coupled structures with hybrid uncertainties. Acta Mech. 231, 1621–
(2020).f (2020). 33. Stabile, L., Scungio, M., Buonanno, G., Arpino, F. & Ficco, G. Airborne particle emission of a commercial 3d printer: the effect of
fil
t
t
i l
d
i ti
t
t
I d
Ai 27 398 408 (2017) 33. Stabile, L., Scungio, M., Buonanno, G., Arpino, F. & Ficco, G. Airborne particle emission of a commercial 3d printer: the effe
filament material and printing temperature. Indoor Air 27, 398–408 (2017).i i
p
g
p
,
(
)
34. Melenka, G. W., Schofield, J. S., Dawson, M. R. & Carey, J. P. Evaluation of dimensional accuracy and material properties of the
makerbot 3d desktop printer. Rapid Prototyp. J. 21, 618–627 (2015). https://doi.org/10.1038/s41598-021-94520-x Scientific Reports | (2021) 11:15221 | Additional information Correspondence and requests for materials should be addressed to T.C. Correspondence and requests for materials should be addressed to T.C. Reprints and permissions information is available at www.nature.com/reprints. Reprints and permissions information is available at www.nature.com/reprints. Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and
institutional affiliations. Open Access This article is licensed under a Creative Commons Attribution 4.0 International
License, which permits use, sharing, adaptation, distribution and reproduction in any medium or
format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the
Creative Commons licence, and indicate if changes were made. The images or other third party material in this
article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the
material. If material is not included in the article’s Creative Commons licence and your intended use is not
permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from
the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. © The Author(s) 2021 https://doi.org/10.1038/s41598-021-94520-x Scientific Reports | (2021) 11:15221 |
|
https://openalex.org/W2956310934
|
https://europepmc.org/articles/pmc6621949?pdf=render
|
English
| null |
What topics should we teach the parents of admitted neonates in the newborn care unit in the resource-limited setting - a Delphi study
|
Maternal health, neonatology and perinatology
| 2,019
|
cc-by
| 5,939
|
Maternal Health, Neonatology,
and Perinatology Maternal Health, Neonatology,
and Perinatology Musabyemungu et al. Maternal Health, Neonatology, and Perinatology
(2019) 5:11 https://doi.org/10.1186/s40748-019-0106-8 Abstract Background: In resource-limited settings, such as Rwanda, health care profession (HCP) to neonate ratios are low,
and therefore caregivers play a significant role in providing care for their admitted neonates. To provide such Family
Integrated Care, caregivers need knowledge, skills, and confidence. The objective of this study was to identify consensus
from key stakeholders regarding the priority topics for a “parental neonatal curriculum.” Methods: A three-round Delphi-study was conducted. During Round-1, face-to-face interviews were undertaken and
responses coded and categorized into themes. In Round-2, participants were presented with Round-1 feedback and
asked to provide additional topics in respective themes. In Round-3, respondents were asked to rank the importance of
these items using a 9-point Likert scale. Results: Ten, 36 and 40 stakeholders participated in Rounds-1, −2 and −3 respectively, including parents, midwives,
nurses and physicians. Twenty and 37 education topics were identified in Rounds-1 and -2 respectively. In Round-3 47
of the 57 presented outcomes met pre-defined criteria for inclusion in the “parental neonatal curriculum.” Conclusion: We describe a “parental neonatal curriculum,” formed using robust consensus methods, describing the core
topics required to educate parents of neonates admitted to a newborn care unit. The curriculum has been developed in
Rwanda and is relevant to other resource-limited settings. Keywords: Education, Caregiver, Infant, newborn, Developing countries to substantially reduce newborn mortality [5]. Monitoring
and evaluation of interventions and care programs are
vital to identify effective interventions [6]. The Rwandan
Ministry of Health includes a Neonatal Working Group to
implement such change nationally and data monitoring is
undertaken using the Integrated Health Management
Information System (HMIS) [7]. © The Author(s). 2019 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and
reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to
the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver
(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. What topics should we teach the parents of
admitted neonates in the newborn care
unit in the resource-limited setting -
a Delphi study Jean Aime Musabyemungu1,2*
, Alice Willson4
, Sean Batenhorst5
, James Webbe6
and
Peter Thomas Cartledge2,3 Jean Aime Musabyemungu1,2*
, Alice Willson4
, Sean Batenhorst5
, James Webbe6
and
Peter Thomas Cartledge2,3 Jean Aime Musabyemungu1,2*
, Alice Willson4
, Sean Batenhorst5
, James Webbe6
and
Peter Thomas Cartledge2,3 RESEARCH ARTICLE Open Access What topics should we teach the parents of
admitted neonates in the newborn care
unit in the resource-limited setting -
a Delphi study Introduction The majority of neonatal deaths occur in two regions of
the world: 39% in sub-Saharan Africa and 38% in Southern
Asia with 99% of neonatal mortality being found in the
resource-limited setting [1]. Worldwide neonatal mortality
has declined slower than other rates of under-5 mortality
[2, 3]. Rwanda is committed to meeting Sustainable Deve-
lopment Goal (SDG) 3.2.2: ending preventable deaths of
newborns and children under 5 years of age by 2030 [4]. In order to achieve this care facilities in low-income coun-
tries (LICs) should deliver proven, effective interventions In resource-limited settings, such as Rwanda, nurse to
neonate ratios are low. Family Integrated Care (FICare) is
frequently employed as a necessity, integrating parents as
primary caregivers of their sick newborns [8, 9]. Care pro-
vided by parents to their admitted neonate is dependent
upon knowledge, practice, and confidence of the parents
regarding neonatal care and may go on to determine the
neonate’s health status and length of neonatal admission. * Correspondence: musjaime@gmail.com
1University of Rwanda, Kigali, Rwanda
2University Teaching Hospital of Kigali (CHUK), Kigali, Rwanda
Full list of author information is available at the end of the article * Correspondence: musjaime@gmail.com
1University of Rwanda, Kigali, Rwanda
2University Teaching Hospital of Kigali (CHUK), Kigali, Rwanda
Full list of author information is available at the end of the article Page 2 of 9 (2019) 5:11 Musabyemungu et al. Maternal Health, Neonatology, and Perinatology It is possible that in the resource-limited setting that there
is a contribution to morbidity and mortality due to an in-
adequate parental understanding of neonatal illness and
the care that these newborns require [10]. Therefore,
equipping parents with proper knowledge of essential neo-
natal care could contribute to improving the outcomes of
these neonates [10–13]. prognosis where participation could be distressing for
the participant were excluded along with parents
who were themselves under 18 years-of-age. Parents
were recruited at two newborn care units of the
University Teaching Hospital Kigali (CHUK), and
Muhima District Hospital (MDH). Due to the
transient nature of parents at the two sites, the
parent participants were different in each Round of
the Delphi study. Convenience sampling was
employed at the clinical sites. The admission of a sick neonate is a stressful period for
parents [14, 15] and so education of parents is paramount
to not only improve the quality of care they provide but
also to reduce the stress of caregiving and enhance confi-
dence at the point of discharge [16, 17]. As the number of
HCPs is low, priority should be given to the key topics to
ensure maximum benefit for the neonate without over-
burdening HCPs with educational roles. Both units are found in Kigali, the capital city of
Rwanda. CHUK is a tertiary level hospital with the new-
born unit has approximately 560 admissions and caters
for 20–30 infants every day, with three Kangaroo
Mother Care (KMC) spaces. The obstetric department is
a referral unit and the principal site for approximately
2000 high-risk deliveries per year [22]. MDH is a district
hospital, located in Kigali city, and serves approximately
1 million people. The hospital has only two major
departments: obstetrics & gynecology and pediatrics
with
neonatology
and
is
responsible
for
approxi-
mately 15,000 deliveries per year. The MDH neonatal
unit includes 25 cot spaces and eight KMC spaces. Both neonatal units would be considered a level II
by USA standards [23] and level I by UK standards
[24], providing simple therapies such as CPAP and intra-
venous fluids, without mechanical ventilation or total par-
ental nutrition. There are no admission weight cut-offs,
and standard practice requires a weight of 1.8 kg before
discharge [22]. Scope Group 2 - Expert stakeholders: We defined an “expert”
as professionals who had experience in clinical care for
neonates and their families in a resource-limited
setting, such as Rwanda. These experts were drawn
from the following: (i) Nurses and midwives at the two
clinical sites; (ii) Rwandan clinicians and residents
working in Rwanda in pediatric and neonatal care who
were identified via the pediatric academic faculty at
the University of Rwanda; (iii) Members of the
Rwandan Ministry of Health (MoH) Neonatal
Working Group (NWG) including pediatricians,
nurses and midwives, identified through the chair of
the NWG; (iv) General Practitioners (clinicians
working in district hospitals) identified through the
class-representatives at the University of Rwanda;
(v) Non-Rwandan, international pediatricians and
neonatologists with experience of working in
Rwanda through the Human Resources for Health
(HRH) program [25] identified from the Ministry
of Health (MoH) database of HRH faculty. We
communicated with the expert stakeholders by
e-mail or via visiting the two clinical sites. This “parental neonatal curriculum” describes the core
topics to educate parents of neonates admitted to a neo-
natal unit in Rwanda, and would also be applicable in
other resource-limited settings. Study design This was a three-round Delphi study. Delphi methods use
sequential “rounds,” with controlled feedback between
rounds to build consensus from a group of experts [21]. The Delphi method is useful in situations where individual
opinions and judgments need to be considered and com-
bined to answer an incomplete state of knowledge. The
process was “fully anonymized,” that is participants did
not know the identities of the other individuals in the
group, nor did they know the specific answers that any
other individual had given. Aims To identify consensus from key stakeholders regarding
the priority topics for a “parental neonatal curriculum.” Parental education could include topics such as imme-
diate and exclusive breastfeeding, hand washing, manage-
ment of hypothermia, hygienic cord cleaning, recognition
of danger signs for neonatal illness and kangaroo mother
care (KMC) for low birthweight (LBW) neonates [2, 18]. It
has been shown that parental education is associated with
improved neonatal care practices in well, non-admitted
neonates [19, 20]. Currently there is no consensus about
what topics parents of sick, admitted, neonates should be
taught in this setting. Results No deviation in the original study protocol was required. Reporting of this study is per the COS-STAR checklist
for Delphi studies [29]. Participants We recruited two groups: Group 1 - Parents: Parents of admitted neonates were
eligible for inclusion. Parents of neonates with a poor Page 3 of 9 (2019) 5:11 Page 3 of 9 Musabyemungu et al. Maternal Health, Neonatology, and Perinatology Round-1 (oral open-questions) Face-to-face interviews were employed to build an initial
draft list of the “parental neonatal curriculum” topics. Two
open questions were posed (see Additional file 1) for par-
ticipants to describe the topics. As topics were identified
using interviews there was no word limit on responses. The questions were asked verbally with responses col-
lected by the PI using field notes. No voice recordings of
the interviews were undertaken. Parents responses were
then translated by the PI. HCPs responded in English. The
responses were then coded, and summarised in Microsoft
Excel by the PI (JAM) and supervising consultant (PC). Consensus for inclusion in Round-2, was pre-defined as
any topic suggested by any one participant. The initial list
of topics was categorized into five domains. Round-3 (closed-questions) Round 3 (closed questions)
Feedback was given to participants with the items from
Round-1 and -2 being combined in a single list, and by
presenting each topic with feedback in the form of a per-
centage of participants who had suggested it. These
items were presented to parents and expert stakeholders
who were asked to grade the importance of the topics
using a 1–9 point Likert scale as described by the
GRADE development group [27, 28]. The data-collector
(JAM) presented the list to parents and was available to
clarify any items that parents did not understand. Consensus for inclusion in the final “parental neonatal
curriculum” was pre-defined as items with greater than
70% of participants scoring 7–9 (important) AND less
than 15% of participants scoring 1–3 (not important) [28]. Questionnaire development The questionnaires were designed specifically for the
purposes of this study. The questionnaires and feedback
were translated for parents into Kinyarwanda, the single
unifying language of Rwanda, by the Principal Investi-
gator (JAM). Expert stakeholders (Group-2) completed
the questionnaire in English. All questionnaires were
piloted for understanding before use. Paper question-
naires (face-to-face) were administered at the clinical
sites for parents, nurses and midwives (CHUK and
MDH) and therefore this group of stakeholders did not
require internet access. Electronic questionnaires (Google
Forms®) were sent by email to expert stakeholders
(Group-2) found outside of the clinical sites. Participants Ten, 36, and 40 participants took part in Rounds-1, −2
and −3 respectively (Table 1 and Fig. 1). The overall
response rate was 91, 38 and 43% respectively which far
exceeding our predicted response rate. The overall re-
sponse rate was 100% amongst parents and 35% (59/171)
in the experts. Expert stakeholders were from four
countries; Rwanda, USA, United Kingdom, and Tanzania. Sample size asked to add any additional topics that they felt were
missing, within that domain, and should be added to
the curriculum (see Additional file 2: for questionnaire). Parental responses were translated to English by the PI. HCPs responded in English. Responses were then coded
and analyzed in Microsoft Excel. Duplicate items from
Round-1 were removed. Consensus for new items to be
included in Round-3 was pre-defined as any single
topic that was given by any one participant. p
In Round-1 we aimed to undertake face-to-face interviews
with 10 participants. For Rounds-2 and -3, we aimed to
gain responses from a minimum of 15 respondents, in
each round, which is considered the required number for
achieving consensus in Delphi studies [26]. Group-2
response rate was predicted to be 10%, therefore, invites
were sent to 80 potentially eligible participants. Non-
participation in Round-2 did not exclude participation in
Round-3. New participants in Group 2 were not added
between Round-2 and -3. The exception to this were
nurses and midwives at the clinical sites who were
recruited opportunistically at the clinical sites and com-
pleted paper rather than online questionnaires. Correlation of importance of topics between stakeholder
groups To assess for overall correlation in opinion between the
three stakeholder groups comparison of the mean scores
of each topic was undertaken using linear regression and
Pearson’s correlation (R). The importance of each indivi-
dual topic was categorized into three levels of importance,
namely 7–9 (important), 4–6 (intermediate) and 1–3 (not
important) and then each individual item was compared
between subject groups (clinicians, nurses, caregivers)
using Chi-squared. Each item was color coded for import-
ance with green representing high importance and red
reflecting low importance. This allows for a visual com-
parison between the stakeholder groups. Consensus process
Participants took part in three rounds of surveys. Round-1 In Round-1, during face-to-face interviews, the partici-
pants generated 20 education topics (Table 2). After cod-
ing, the topics were categorized into five themes; (i)
Topics at admission; (ii) General neonatal care; (iii)
Feeding; (iv) Cleanliness and Hygiene, and (v) Topics to
be taught at the discharge period (Table 2). During
Round-1 each topic was described by a mean of 4.0 parti-
cipants (SD ± 2.0). Only three of the 20 (15.0%) topics
were suggested by only one of the participants. Round-2 Round-2
In Round-2 the participants generated 37 additional topics
(Table 2) and these were again classified in the themes
identified in Round-1. The 37 new topics were each
described by a mean of 2.8 (±2.6) participants. Sixteen of
the topics were suggested by only one of any of the stake-
holders taking part in the Round (i.e. they were alone in
suggesting that topic). Round-2 (free-text open-questions) Feedback from Round-1 was given to participants, with all
the topics generated in Round-1 being presented to partic-
ipants in the questionnaire. The items were presented
within each individual domain and participants were then Musabyemungu et al. Maternal Health, Neonatology, and Perinatology (2019) 5:11
Page 4 of 9 Musabyemungu et al. Maternal Health, Neonatology, and Perinatology (2019) 5:11 Page 4 of 9 Table 1 Baseline characteristics of participants of Rounds 1–3
Round-1 (n = 10)
Round-2 (n = 36)
Round-3 (n = 40)
All
Response rate
All
10/11 (90.9%)
36/94 (38.2%)
40/93 (43.0%)
HCPs
5/6 (83.3%)
24/82 (29.3%)
30/83 (36.1%)
Parents
5/5 (100%)
12/12 (100%)
10/10 (100%)
Questionnaire administration
Electronic
NA
22 (61.1%)
26 (65.0%)
Paper
14 (38.9%)
14 (35.0%)
Role
Parent
5 (50.0%)
12 (33.0%)
10 (25%)
Pediatricians
3 (30%)
13 (36.1%)
20 (50%)
General Practitioner
0 (0.0%)
6 (17.0%)
4 (10.0%)
Nurses and midwives
2 (10.0%)
5 (13.9%)
6 (53.0%)
Age
20–29
5 (50.0%)
11 (30.6%)
12 (30.0%)
30–39
4 (40.0%)
21 (58.3%)
23 (57.5%)
> 40
1 (10.0%)
4 (11.1%)
1 (2.5%)
Gender
Male
1 (10.0%)
17 (47.2%)
16 (40.0%)
Female
9 (90.0%)
19 (52.8%)
24 (60.0%)
HCPs
HCPs Main place of work
Rwanda
5 (100%)
21 (88.0%)
26 (87.0%)
USA
0 (0%)
2 (8.0%)
2 (7.0%)
United Kingdom
0 (0%)
0 (0%)
2 (7.0%)
Tanzania
0 (0%)
1 (4.0%)
0 (0%)
HCPs Years of experience
Mean
9.3 (±8.4)
5.2 (±4.9)
6.2 (±5.3)
HCPs - How often treating neonate
Never or rarely
0 (0%)
4 (15.3%)
0 (0%)
Sometimes
1 (20.0%)
2 (7.7%)
4 (13.3%)
Frequently or very frequently
4 (80.0%)
20 (76.9%)
26 (86.7%)
Parents
Parent Hospital
MDH
3 (60.0%)
9 (75.0%)
4 (40.0%)
CHUK
2 (40.0%
3 (25.0%)
6 (60.0%)
Parent Social economic status
1–2 (Low)
1 (20.0%)
6 (50.0%)
3 (30.0%)
2–4 (High)
4 (80.0%)
6 (50.0%)
7 (70.0%)
Parental Residence
Urban
4 (80.0%)
12 (100%)
8 (80.0%)
Rural
1 (20.0%)
0 (0%)
2 (20.0%)
Parent Education
No education or primary
3 (60.0%)
6 (50.0%)
4 (44.4%)
Secondary or Higher
2 (40.0%)
6 (50.0%)
5 (55.6%) Table 1 Baseline characteristics of participants of Rounds 1–3 There was no mechanism to measure any challenges with
access
in
stakeholders
receiving
the
questionnaire
electronically. Parents were a key group within our stake-
holders, representing 50, 33 and 25% of participants in
Rounds-1, −2 and −3 respectively (Table 1). Round-2 Round-3 In Round-3 the 20 and 37 topics from Rounds-1 and
-2 respectively were combined, and the 57 items
were
ranked
for
importance
by
the
participants. Forty-seven (84%) topics met the pre-defined consen-
sus criteria to be included in the “parental neonatal
curriculum” (Table 2). Musabyemungu et al. Maternal Health, Neonatology, and Perinatology (2019) 5:11
Page 5 of 9 Musabyemungu et al. Maternal Health, Neonatology, and Perinatology (2019) 5:11 Page 5 of 9 Musabyemungu et al. Maternal Health, Neonatology, and Perinatology (2019) 5:11 Fig. 1 Study flow-chart Fig. 1 Study flow-chart Parental neonatal curriculum quantity” and “hand washing” scoring the highest in each
of these categories respectively. Finally, there were a series
of topics that were considered to be important at the point
of discharge, such as “follow up planning”. Nineteen of the
topics in the curriculum were also specific to caregiving
(e.g., feeding through a nasogastric tube). The topics were categorized into five domains represen-
ting the aspects of care of admitted newborns (Table 1
and Additional file 1). All the topics (except discharge
topics) within our curriculum would be essential for any
parent who is providing FICare of an admitted newborn
in this setting. Topics on admission were generally about
providing an “induction” for parents to the newborn care
unit and explaining the reason for admission. Topics were
then divided into the domains of “general care”, “feeding”
and “hygiene” with “Kangaroo mother care”, “feeding Correlation between groups There was a moderate correlation in the importance of
items between the three groups of stakeholders with Pear-
son’s R ranging between 0.52–0.57, p < 0.001 (Fig. 2). In Table 2 Sections of the topics for the curriculum
Sections
Round-1
Round-2
Round-3
Round-3
Total number
of items
New items from
participants
Total number of items
presented in Round-3
Consensus criteria met
Topics at admission
6
8
14
8 (61.5%)
General care
4
8
12
10 (83.3%)
Feeding
6
7
13
11 (84.6%)
Cleanliness and hygiene
2
2
4
4 (100%)
Topics at discharge
2
12
14
14 (100%)
Total education topics
20
37
57
47 (83.9%) Musabyemungu et al. Maternal Health, Neonatology, and Perinatology (2019) 5:11
Page 6 of 9 Musabyemungu et al. Maternal Health, Neonatology, and Perinatology (2019) 5:11 Musabyemungu et al. Maternal Health, Neonatology, and Perinatology Page 6 of 9 Fig. 2 Correlation of Education topics between participants. R = Pearson’s R correlation ig. 2 Correlation of Education topics between participants. R = Pearson’s R correlation discharge. This curriculum has been developed using ro-
bust Delphi-consensus techniques. Table 3 items have been presented with color coding to
aid recognition of items with discordance in the opinion
of stakeholders. Healthcare professional competency and knowledge Healthcare professional competency and knowledge
Many parents will arrive on a neonatal unit with poor
education levels and this can be a barrier to providing
care [34]. In our stakeholder groups approximately 50%
of participants had only primary or no formal education. It is also important to consider the level of knowledge
and competencies in the healthcare professionals who
may be educating parents on how to care for their sick,
admitted, newborn. Ensuring that staff are well trained
and have ongoing training to maintain competencies is
essential for their own ability to care for neonates and to
educate and support parents [34–36]. Levels of agreement on curriculum items Three groups of major stakeholders have been included
here to gain a balanced curriculum to reflect the needs
and wishes of each group of stakeholders. There was a
moderate correlation in the importance of items be-
tween the three groups of stakeholders with Pearson’s R
ranging between 0.52–0.57, p < 0.001 (Fig. 2). However,
there were some topics where opinions varied markedly
(Table 3). For example, parents had a strong wish for
education on family planning which was not shared by
nurses and clinicians. Even though there was some
variation in opinions between stakeholder groups, only
four of these were significantly different, and these four
could also be explained by a large number of topics
being tested without Bonferroni correction. Discussion The admission of sick or preterm neonates is traumatic
and stressful for parents; long-term it can cause impaired
bonding and symptoms of post-traumatic stress disorder,
affecting neonatal outcomes such as feeding [30]. FICare
is a model of care that aims to reduce these adverse effects
by integrating parents as primary caregivers of their sick This research project has identified the priority topics to
be included in a “parental neonatal curriculum” for par-
ents of admitted neonates in a resource-limited setting. The curriculum includes topics relating to admission,
general care, feeding, cleanliness and hygiene, and Musabyemungu et al. Maternal Health, Neonatology, and Perinatology Page 7 of 9 Page 7 of 9 (2019) 5:11 Table 3 All education topics presented in Round-3 newborns [8]. However, their integration needs to be
underpinned by peer support and education; the Family-
Led Care Model (FLCM) in Malawi is an excellent
example of this [31]. FLCM was developed to improve
facility- and home-based care of preterm/LBW newborns. This model enhances the skills of providers and the
quality of care within KMC units, empowering families to
directly participate in the care of their preterm/LBW
newborn while still in the facility and with access to
trained providers [32]. In Pakistan, it has been shown that
it is possible to involve mothers in the active care of their
very low birthweight infants before discharge and that this
may translate into earlier discharge from the hospital
without an increase in short term complications and
readmission [33]. This FICare approach provides a cost-
effective strategy for newborn units in resource-limited
settings providing parents have been adequately educated. Table 3 All education topics presented in Round-3
Chi squared p value Informed consent A consent form was written specifically for this student and piloted for
understanding. Strengths The strengths of this study include a high response rate
from parents and clinicians and the broad experience of
the HCPs who participated in this study. We followed a
robust consensus methodology and have reported this
work fully. Confidentiality
N
id
ifi bl y
No identifiable patient data were collected during the project. No identifiable patient data were collected during the project. Future research in this area The study was undertaken as the undergraduate thesis of the Principal
Investigator (JAM). JAM was supported and supervised by PC and AW. JAM
and PC conducted the data-collection. JAM and PC undertook the analysis. AW, JW and SB contributed to the interpretation of the results. All authors
were significant contributors in writing the manuscript. All authors read and
approved the final manuscript. Questions for further research include: what are the best
methods to implement a curriculum of education for
these parents, e.g. videos, group workshops, written
literature, expert mothers, etc. We also did not investi-
gate which parents should receive this curriculum to
gain the maximum benefit. Education should enhance
care, being implemented as part of a package to upscale Risk to subjects No significant physical, social, legal or financial risks were identified. Emotional
risks included parental anxiety from considering the care of their sick neonate:
The research participants were reminded that they held their right to withdraw
from the research or limit their participation at any point. Limitations There is a lack of previous studies published on this
particular subject to draw comparison. In Round-1 the
face-to-face interviews were not audio-recorded. The
purpose of Round-1 was not to identify rich theory (such
as found in qualitative research), but rather to identify a
preliminary list of topics. Audio-recording may have CChi-squared p-value Musabyemungu et al. Maternal Health, Neonatology, and Perinatology Page 8 of 9 (2019) 5:11 Page 8 of 9 made participants reluctant to give full responses, fearful
that they are being recorded. However, it is possible
some topics were not documented in the field notes. The generalization of the study findings is limited
because data collection occurred at just two hospital
sites and in one geographical location. Another possible
source of bias was acquiescence bias, the tendency of
the participant to agree with statements or influential
panel members, which we avoided in the initial rounds
by explicitly avoiding the questions of agree/disagree
and in later rounds by conducting a fully anonymized
survey with no direct interaction between participants. Stakeholders tended to report that all the topics were
“important”, resulting in only a small number of topics
being excluded from the final curriculum. This could be
explained by “respondent fatigue” in having to review 56
items for importance. care. Therefore, future research could potentially investi-
gate the formal implementation of FICare in this setting,
with parental education being a part of this package of
care, and whether this is effective and cost-effective. Once implemented quantitative investigation of changes
in care would be required as evidence in this setting is
lacking. This may involve regular data collection through a
Neonatal Registry [22] to allow for measures of change and
loops of Quality Improvement [36]. Qualitative research
would also be important to identify how individual popula-
tions want to receive education and which aspects they find
most beneficial for caring for their admitted newborn. Conclusion We have described a “parental neonatal curriculum,”
formed using robust consensus methods. Greater improve-
ment in neonatal care practices is essential if neonatal
mortality reduction is to be achieved in resource-limited
settings where the burden of disease is found. One step in
achieving this could include the use of proven low-cost
interventions such as FICare, based on a foundation of
effective parental education. Additional file 1: Parental neonatal curriculum. (DOCX 20 kb)
Additional file 2: Questions posed to participants. (DOCX 18 kb) Any newborn care unit wanting to use the neonatal cur-
riculum described here would be encouraged to tailor it
their own patient population and their care needs. Our
curriculum of topics was extensive, with 49 itemswhich
may be beyond the capacity of a particular unit. The
topics found in each domain of our curriculum have
been ordered by “importance” given by the stakeholders
so units may want to choose items which they feel are of
“High”, “Medium” and “Low” priority based on their
populations needs and on the capacity of their HCPs. Each subgroup of patients may also have distinct educa-
tional needs: the multiparous mother of an extremely
preterm newborn may have very different educational
needs to a primiparous mother of a near-term infant. There is no “one size fits all” educational package. Once
parents have been given education on a curriculum item
(e.g. nasogastric feeding) a competent HCP should assess
the parental knowledge and skills before the specific task
is undertaken by a caregiver. Abbreviations CHUK: University Teaching Hospital of Kigali; FICare: Family Integrated Care;
HCPs: Health Care Professionals; HRH: Human Resources for Health;
KMC: Kangaroo Mother Care; LBW: Low Birthweight; MoH: Ministry of Health;
NMR: Neonatal Mortality Rate No incentives were provided to the participant. No incentives were provided to the participant. Application of the findings Additional file 1: Parental neonatal curriculum. (DOCX 20 kb)
Additional file 2: Questions posed to participants. (DOCX 18 kb) Incentives for subjects No incentives were provided to the participant. References Cited 28 May 2019. Available from: https://hmis.moh.gov.rw/. 7. Rwanda Ministry of Health - Integrated Health Management Information
System (HMIS). Cited 28 May 2019. Available from: https://hmis.moh.gov.rw/. 8. Gale C. Comment family integrated care for very preterm infants: evidence
for a practice that seems self-evident ? Lancet Child Adolesc. 2018;2:1–2. 34. Martinez AM, Khu DTK, Boo NY, Neou L, Saysanasongkham B, Partridge JC. Barriers to neonatal care in developing countries: parents’ and providers’
perceptions. J Paediatr Child Health. 2012;48:852–8. 9. National Institute of statistics. Rwanda Demographic and Health Survey. Rwanda: National Institute of Statistics; 2015. https://dhsprogram.com/pubs/
pdf/FR316/FR316.pdf. 9. National Institute of statistics. Rwanda Demographic and Health Survey. Rwanda: National Institute of Statistics; 2015. https://dhsprogram.com/pubs/
pdf/FR316/FR316.pdf. 35. Namazzi G, Waiswa P, Nakakeeto M, Nakibuuka VK, Namutamba S, Najjemba
M, et al. Strengthening health facilities for maternal and newborn care:
experiences from rural eastern Uganda. Glob Health Action. 2015;1:1–8. 10. Weiner EA, Billamay S, Partridge JC, Martinez AM. Antenatal education for
expectant mothers results in sustained improvement in knowledge of
newborn care. J Perinatol. 2011;31:92–7. 36. Zaka N, Alexander EC, Manikam L, Norman ICF, Akhbari M, Moxon S,
et al. Quality improvement initiatives for hospitalised small and sick
newborns in low- and middle-income countries: a systematic review. Implement Sci. 2018;13:1–21. 11. Amolo L, Irimu G, Njai D. Knowledge of postnatal mothers on essential
newborn care practices at the Kenyatta national hospital: a cross sectional
study. Pan Afr Med J. 2017;28:1–7. 12. Meseka LA, Mungai LW, Musoke R. Mothers’ knowledge on essential
newborn care at juba teaching hospital, South Sudan. South Sudan Med J. 2017;10:56–9. Funding Support provided by OpTin (Overseas Partnering and Training Initiative), a UK
charity based at the Leeds Teaching Hospital NHS Trust. The funder played no Page 9 of 9 Page 9 of 9 Page 9 of 9 (2019) 5:11 Musabyemungu et al. Maternal Health, Neonatology, and Perinatology role in the design of the study and collection, analysis, and interpretation of
data or in writing the manuscript. 16. Morey JA, Gregory K. Nurse-led education mitigates maternal stress and
enhances knowledge in the NICU. Am J Matern Nurs. 2012;37:182–91. 17. Khee Loo K, Espinosa M, Tyler R, Howard J. Using knowledge to cope with
stress in the NICU: how parents integrate learning to read the physiologic
and behavioral cues of the infant. Neonatal Netw. 2003;22:31–7. The authors declare that they have no competing interests. The authors declare that they have no competing interests. 23. American Academy of Pediatrics. Levels of neonatal care. Pediatrics. 2012;130:587–97. 23. American Academy of Pediatrics. Levels of neonatal care. Pediatrics. 2012;130:587–97. Author details
1 24. NHS networks. UK definitions of levels of neonatal care. https://www. networks.nhs.uk/nhs-networks/staffordshire-shropshire-and-black-country-
newborn/documents/StandardsAssessmentDefinitions_000.pdf. 1University of Rwanda, Kigali, Rwanda. 2University Teaching Hospital of Kigali
(CHUK), Kigali, Rwanda. 3Rwanda Human Resources for Health (HRH)
Program, Yale University (USA), Kigali, Rwanda. 4Royal College of Paediatrics
and Child Health, UNICEF neonatal programme, Kigali, Rwanda. 5University of
Wyoming, Laramie, Wyoming, USA. 6Imperial College London, London, UK. 25. Nuthulaganti T, Umubyeyi B, Sc MN, Nyemazi JP, Uwayezu A, Sc MN,
et al. The human resources for health program in Rwanda - a new
partnership. 2013; 26. Hsu C, Sandford B. The delphi technique: making sense of consensus. Pract
Assess Res Eval. 2007;12:1–8. 26. Hsu C, Sandford B. The delphi technique: making sense of consensus. Pract
Assess Res Eval. 2007;12:1–8. Received: 5 April 2019 Accepted: 25 June 2019 Received: 5 April 2019 Accepted: 25 June 2019 27. GRADE. Handbook for grading the quality of evidence and the strength of
recommendations using the GRADE approach 2013 [cited 4 Jan 2018]. Available from: http://gdt.guidelinedevelopment.org/app/handbook/
handbook.html#h.1i2bwkm8zpjo. Consent for publication
Not applicable. Consent for publication
Not applicable. 21. Dalkey N, Helmer O. An experimental application of the Delphi method to
the use of experts - report prepared for the United States air force; 1962. 22. Choi J, Urubuto F, Dusabimana R, Kumwami M, Cartledge PT, Agaba F, et a
Establishing a neonatal database in a tertiary hospital in Rwanda – an
observational study. Paediatr Int Child Health. 2019;00:1–10. References 1. Lawn JE, Cousens S, Zupan J. Neonatal survival 14 million neonatal deaths:
when? Where? Why? Lancet. 2005;365:891–900. 1. Lawn JE, Cousens S, Zupan J. Neonatal survival 14 million neonatal deaths:
when? Where? Why? Lancet. 2005;365:891–900. 28. Williamson PR, Altman DG, Bagley H, Barnes KL, Blazeby JM, Brookes ST,
et al. The COMET handbook: version 1.0. Trials. 2017;18:1–50. 2. UNICEF. Committing to child survival: a promise renewed. Progress report. 2014. http://files.unicef.org/publications/files/APR_2014_web_15Sept14.pdf. 2. UNICEF. Committing to child survival: a promise renewed. Progress report. 2014. http://files.unicef.org/publications/files/APR_2014_web_15Sept14.pdf. 29. Kirkham JJ, Gorst S, Altman DG, Blazeby JM, Clarke M, Devane D, et al. Core
outcome set–standards for reporting: the COS-STAR statement. PLoS Med. 2016;13:1–11. 3. Lawn JE, Blencowe H, Oza S, You D, Lee ACC, Waiswa P, et al. Every
newborn: Progress, priorities, and potential beyond survival. Lancet. 2014;384:189–205. 30. Pierrehumbert B, Nicole A, Muller-Niz C, Forcada-Guex M, Ansermet F. Parental post-traumatic reactions after premature birth: implications for
sleeping and eating problems in the infant. Arch Dis Child Fetal Neonatal
Ed. 2003;88:400–4. 4. UNDP. SDG 3: ensure healthy lives and promote wellbeing for all at all
ages. Cited 21 Feb 2019. Available from: https://sustainabledevelopment. un.org/sdg3. 4. UNDP. SDG 3: ensure healthy lives and promote wellbeing for all at all
ages. Cited 21 Feb 2019. Available from: https://sustainabledevelopment. un.org/sdg3. 5. English M. Designing a theory-informed , contextually appropriate
intervention strategy to improve delivery of paediatric services in Kenyan
hospitals. Implement Sci. 2013;8:1. 31. Every Preemie. Family-led care package - Malawi. Washington DC. [cited 15
Mar 2019]. Available from: http://www.everypreemie.org/wp-content/
uploads/2017/05/Family-Led-Care-Malawi-Package.pdf 6. Maina M, Aluvaala J, Mwaniki P, Tosas-Auguet O, Mutinda C, Maina B, et al. Using a common data platform to facilitate audit and feedback on the
quality of hospital care provided to sick newborns in Kenya. BMJ Glob
Health. 2018;3:e001027. 32. Every-Premie. Every Preemie-SCALE (scaling, catalyzing, advocating, learning,
and evidence-driven). Cited 28 May 2019 . Available from: https://www. everypreemie.org/malawi/family-led-care/. 33. Bhutta ZA, Khan I, Salat S, Raza F, Ara H. Reducing length of stay in hospital
for very low birthweight infants by involving mothers in a stepdown unit:
an experience from Karachi (Pakistan). BMJ. 2004;329:1151–5. 33. Bhutta ZA, Khan I, Salat S, Raza F, Ara H. Reducing length of stay in hospital
for very low birthweight infants by involving mothers in a stepdown unit:
an experience from Karachi (Pakistan). BMJ. 2004;329:1151–5. 7. Rwanda Ministry of Health - Integrated Health Management Information
System (HMIS). Availability of data and materials The datasets generated and/or analyzed during the current study are not
publicly available due to requiring consent from the approving ethical
committee to share but are available from the corresponding author on
reasonable request and approval from the relevant ethics board. 18. UNICEF. The state of world’s children - report; 2009. p. 168. [cited 28 May
2019]. Available from: https://www.unicef.org/sowc09/docs/SOWC09-
FullReport-EN.pdf. 19. Nnebue C, Duru C, Uwakewe K, Ifeadike C, Anyanwu B, Adinnus K, et al. Neonatal care – what do mothers in a rural Nigerian community know and
practice ? J Neonatal Perinat Med. 2016;9:27–31. Ethics approval and consent to participate The CHUK Research and Ethics Committee (REC) and Muhima District
Hospital REC provided ethics review and approval (Ref: EC/CHUK/621/2018). 20. Shrestha S, Adachi K, Petrini MA, Dean F, Shrestha S, Care H, et al. Development and evaluation of a newborn care education programme in
primiparous mothers in Nepal. Midwifery. 2016;42:21–8. Publisher’s Note Springer Nature remains neutral with regard to jurisdictional claims in
published maps and institutional affiliations. Springer Nature remains neutral with regard to jurisdictional claims in
published maps and institutional affiliations. 13. Singh K, Brodish P, Haney E. Postnatal care by provider type and
neonatal death in sub-Saharan Africa: a multilevel analysis. BMC Public
Health. 2014;14:1–7. 14. Obeidat HM, Bond EA, Callister LC. The parental experience of having an
infant in the newborn intensive Care unit. J Perinat Educ. 2009;18:23–9. 15. Aagaard H, Hall EOC. Mothers’ experiences of having a preterm infant in the
neonatal care unit: a meta-synthesis. J Pediatr Nurs. 2008;23:26–36.
|
https://openalex.org/W2944369968
|
https://www.bio-conferences.org/10.1051/bioconf/20191408005/pdf
|
English
| null |
More effective use = more lives saved: medical countermeasure response strategies for mass casualty radiological incidents.
|
Bio web of conferences/BIO web of conferences
| 2,019
|
cc-by
| 1,144
|
1 Introduction Strategies for the use of medical countermeasures are vital to ensure maximum lifesaving
during an incident response. Federal budgets for development and acquisition of medical
countermeasures (MCMs) to chemical, biologic, radiological and nuclear (CBRN) threats
are often inadequate to fund infinite sustainment of national stockpiles. In addition,
necessary infrastructure, diagnostic capabilities, operational, and personnel resources can
also be limited or unavailable due to lack of technology or inadequate funds for investment. Thus, federal governments and community emergency medical response and public health
partners must develop strategies to leverage scarce MCMs and other resources to ensure a
mass-casualty response approach to a CBRN incident that efficiently and effectively saves
the most lives. The decision framework approach and MCM response strategies for
radiological incidents will be presented. More effective use = more lives saved: medical
countermeasure response strategies for mass
casualty radiological incidents. Juanita Jones1, and Chad M. Hrdina1
1Assistant Secretary for Preparedness and Response, United States Department of Health and Human
Services, 200 Independent Avenue SW, Washington, DC 20001, United States of America Topic: medical countermeasures and decorporation Topic: medical countermeasures and decorporation 2 Strategic Framework Medical countermeasure response strategies are divided into three major portions with
supporting information: 1) Notification, and 2) Scope of incident, lead to 3) an MCM
response strategy, which is supported with MCM specific information, special
considerations for special populations (e.g., children), regulatory or legal issues or needs for
MCM utilization, as well as useful public messages recommendations, and current clinical
guidelines summary. More effective use = more lives saved: medical
countermeasure response strategies for mass
casualty radiological incidents.
Juanita Jones1, and Chad M. Hrdina1
1Assistant Secretary for Preparedness and Response, United States Department of Health and Human
Services, 200 Independent Avenue SW, Washington, DC 20001, United States of America
T
i
di
l
d d
i More effective use = more lives saved: medical
countermeasure response strategies for mass
casualty radiological incidents. Juanita Jones1, and Chad M. Hrdina1
1Assistant Secretary for Preparedness and Response, United States Department of Health and Human
Services, 200 Independent Avenue SW, Washington, DC 20001, United States of America
Topic: medical countermeasures and decorporation © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons
Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). https://doi.org/10.1051/bioconf/20191408005 https://doi.org/10.1051/bioconf/20191408005 BIO Web of Conferences 14, 08005 (2019)
HEIR 2018 2.2 MCM response strategy In the initial response period, much situational information many not be known, but
decisions will be required to ensure assets, including medical countermeasures are quickly
employed early. As more information becomes available, flexible decision makers can
adjust the medical countermeasure response approach. The MCM response strategy
discusses
target
populations
and
intended
population-based
use
of
medical
countermeasures. Deployment and distribution of medical countermeasures may include
strategies for prophylaxis, mitigation, treatment, of target populations with different
medical countermeasures at different times. The MCM response strategy recommendations
ensure efficient and effective use of medical countermeasures based on confirmed or
estimated factors of the incident and address both adequate and scarce resource strategies. 2.1 Notification and incident scope How we become aware of an incident is an indicator as to the potential affected population
and severity of injuries or illness at the time of notification, which means we can anticipate
the spectrum of casualties resulting from the incident, and make decisions for allocation for
MCMs such as Prussian blue or Ca/Zn-DTPA. The nature of an incident, including factors
such as time, affected population, ability to identify affected population, mode and
magnitude of radiological dispersal are important because they allow responders and
decision makers to identify and characterize the population that has been exposed, or at risk BIO Web of Conferences 14, 08005 (2019) BIO Web of Conferences 14, 08005 (2019)
HEIR 2018 https://doi.org/10.1051/bioconf/20191408005 of exposure. This informs decisions to protect a population at risk, from any or further
exposure, and to implement public health or medical interventions to mitigate or treat injury
from radiation exposure or internal contamination with radionuclides. A very important
aspect of characterizing the magnitude of an incident translates into an estimation as to
whether enough resources are available or there is potential for a shortage. Scoping the
incident is not about identifying all of the potential scenarios, but rather those factors that
differentiate scenarios, such that one would change the strategic approach to response. of exposure. This informs decisions to protect a population at risk, from any or further
exposure, and to implement public health or medical interventions to mitigate or treat injury
from radiation exposure or internal contamination with radionuclides. A very important
aspect of characterizing the magnitude of an incident translates into an estimation as to
whether enough resources are available or there is potential for a shortage. Scoping the
incident is not about identifying all of the potential scenarios, but rather those factors that
differentiate scenarios, such that one would change the strategic approach to response. 3 Radiological incident MCM response strategy Potential for scarce resources following a radiological incident is an important issue that
requires consideration for MCM distribution strategies. We recommend that in a scarce
resources situation, early decorporation remains an important mitigation step that should
not be delayed for confirmatory bioassay to initiate therapy as a means to conserve MCM
resources. Instead, our analyses support an approach where epidemiology and simple
screening can inform pre-emptive administration of MCMs to casualties with suspected
internal contamination, which can save more lives than bioassay confirmation as the
initiation threshold for decorporation. We recommend bioassay for discontinuation of
therapy, and there may be dosing alternatives, as well as alternative thresholds for
discontinuation of therapy, to conserve medical countermeasures. Furthermore,
conservation of resources means establishing appropriate thresholds for target populations,
which could mean palliative care for those too sick to benefit from decorporation. Likewise, response to an incident with sufficient resources availability for the internally
contaminated population should also leverage bioassay for discontinuation of therapy
where thresholds might be set far lower, and screening may include those with less risk. Regardless of resource levels, bioassay is should be prioritized in cases where confirmation
is preferred before initiation of pre-emptive therapy, to reduce high risk for adverse effects
of unnecessary MCM administration (e.g. neonates). 4 Conclusion Deliberative planning the strategic use of medical countermeasures for a variety of
radiological risks means that if a radiological incident were to occur, we are prepared to use
medical countermeasures in the most efficient ways, with the most effective approaches,
such that we maximize the lives-saved. Planning prevents lengthy contemplation during 2 2 https://doi.org/10.1051/bioconf/20191408005 BIO Web of Conferences 14, 08005 (2019) BIO Web of Conferences 14, 08005 (2019)
HEIR 2018 ponse that could impede timely access to medical countermeasures by those who need response that could impede timely access to medical countermeasures by those who need
them. response that could impede timely access to medical countermeasures by those who need
them. 3 3
|
https://openalex.org/W2804914799
|
https://europepmc.org/articles/pmc6005885?pdf=render
|
English
| null |
Net Clinical Benefit of Non-vitamin K Antagonist Oral Anticoagulants for Venous Thromboembolism Prophylaxis in Patients With Cancer: A Systematic Review and Trade-Off Analysis From 9 Randomized Controlled Trials
|
Frontiers in pharmacology
| 2,018
|
cc-by
| 6,983
|
ORIGINAL RESEARCH
published: 12 June 2018
doi: 10.3389/fphar.2018.00575 Edited by:
Sandor Kerpel-Fronius,
Semmelweis University, Hungary Edited by:
Sandor Kerpel-Fronius,
Semmelweis University, Hungary 1 Department of Pharmacy, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China, 2 Department
of Cardiology, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China, 3 Department of Oncology,
State Key Laboratory for Oncogenes and Related Genes, Renji Hospital, School of Medicine, Shanghai Jiaotong University,
Shanghai Cancer Institute, Shanghai, China Reviewed by:
Gaurav Deshpande,
HealthCore, Inc, United States
Sunita Nair,
Independent Researcher, Mumbai,
India Reviewed by:
Gaurav Deshpande,
HealthCore, Inc, United States
Sunita Nair,
Independent Researcher, Mumbai,
India Venous thromboembolism (VTE) is highly prevalent in patients with cancer. Non-vitamin
K antagonist oral anticoagulants (NOACs), directly targeting the enzymatic activity of
thrombin or factor Xa, have been shown to be as effective as and safer than traditional
anticoagulation for VTE prophylaxis in no-cancer patients. However, related studies
that focused on the anticoagulation in cancer patients are lacked, and almost no net
clinical benefit (NCB) analyses that quantified both VTE events and bleeding events
have been addressed in this fragile population. Therefore, we aim to investigate this
issue using a systematic review and NCB analysis. A comprehensive search of Medline,
Embase, and Cochrane Library were performed for randomized controlled trials (RCTs)
that reported the VTE events and major bleeding of NOACs and traditional anticoagulants
in patients with or without cancer. Odds ratios (ORs) and 95% confidence intervals
(CIs) of VTE and bleeding events were calculated using a random-effects model. The
primacy outcome of narrow NCB was calculated by pooling ORs of VTE and major
bleeding, with a weighting of 1.0. Similarly, the broad NCB was calculated by pooling
ORs of VTE and clinically relevant bleeding. Heterogeneity was assessed through I2
test and Q statistic, and subgroup analyses were performed on the basis of different
patients (VTE patients or acutely ill patients), comparators (vitamin-K antagonists or
low-molecular-weight heparin), and follow-up duration (≤6 months or >6 months). Overall, 9 RCTs including 41,454 patients were enrolled, of which 2,902 (7%) were cancer
patients, and 38,552 (93%) were no-cancer patients; 20,712 (50%) were administrated
with NOACs and 20,742 (50%) were administrated with traditional anticoagulants. The
use of NOACs had a superior NCB than traditional anticoagulation in both cancer
patients (OR: 0.68, 95%CI: 0.50-0.85 for narrow NCB; OR: 0.76, 95%CI: 0.61–0.91 *Correspondence:
Li-Wei Wang
liweiwang@shsmu.edu.cn
Zhi-Chun Gu
guzhichun213@163.com †These authors have contributed
equally to this work as first authors. Edited by:
Sandor Kerpel-Fronius,
Semmelweis University, Hungary †These authors have contributed
equally to this work as first authors. Specialty section:
This article was submitted to
Pharmaceutical Medicine and
Outcomes Research,
a section of the journal
Frontiers in Pharmacology
Received: 16 March 2018
Accepted: 14 May 2018
Published: 12 June 2018 Specialty section:
This article was submitted to
Pharmaceutical Medicine and
Outcomes Research,
a section of the journal
Frontiers in Pharmacology Received: 16 March 2018
Accepted: 14 May 2018
Published: 12 June 2018 Yi-Dan Yan 1†, Chi Zhang 1†, Long Shen 2†, Ying-Jie Su 1, Xiao-Yan Liu 1, Li-Wei Wang 3* and
Zhi-Chun Gu 1* Yi-Dan Yan 1†, Chi Zhang 1†, Long Shen 2†, Ying-Jie Su 1, Xiao-Yan Liu 1, Li-Wei Wang 3* and
Zhi-Chun Gu 1* Edited by:
Sandor Kerpel-Fronius,
Semmelweis University, Hungary Data Sources and Searches The present systematic review was conducted in accordance
with standards outlined in the Cochrane Handbook and
the PRISMA Statement for Reporting Systemic Reviews and
was performed according to the priori established protocol
(PROSPERO:
CRD42018089939). We
searched
Medline,
Embase, and Cochrane Library electronic databases to identify
all potential eligible trials from inception to Feb 14th, 2018,
with the following searching strategy: “dabigatran” or “Pradaxa”
or “rivaroxaban” or “Xarelto” or “apixaban” or “Eliquis” or
“edoxaban” or “Savaysa” or “Betrixaban” or “Bevyxxa” or “Non-
vitamin K antagonist oral anticoagulants” or “NOACs” or “direct
oral anticoagulants” or “DOACs” or “novel oral anticoagulants”
or “new oral anticoagulants” or “factor Xa inhibitors” or “factor
IIa inhibitors” AND “venous thromboembolism” or “VTE”
or “pulmonary embolism” or “PE” or “deep vein thrombosis”
or “DVT” AND “clinical trial” or “controlled clinical trial”
or “randomized controlled trials.” References of all pertinent
articles were further scrutinized to ensure that all relevant studies
were identified. Two reviewers (Yi-Dan Yan and Chi Zhang)
independently searched the databases, and all disagreements
were resolved by consulting a third author (Zhi-Chun Gu). Of late years, non-vitamin K antagonist oral anticoagulants
(NOACs), with a predictable dose response and no need for
laboratory monitoring, have been shown to be as effective as and
probably safer than conventional anticoagulation when regarding
VTE treatment and prophylaxis (Schulman et al., 2009, 2013,
2014; Bauersachs et al., 2010; Goldhaber et al., 2011; Büller et al.,
2012, 2013; Agnelli et al., 2013; Cohen et al., 2013; Raskob
et al., 2018), which makes these agents more appealing for long-
term VTE prevention. Whereas, clinical trials of NOACs that
specially aimed at patients with cancer are lacked, and only
a minor proportion of cancer patients (<5%) was involved
in NOACs studies (Schulman et al., 2009, 2014; Bauersachs
et al., 2010; Goldhaber et al., 2011; Büller et al., 2012, 2013;
Agnelli et al., 2013; Cohen et al., 2013; Raskob et al., 2018). For a meaningful analysis, several meta-analysis studies have
been conducted on the focus of this issue by pooling those
5% patients, and demonstrated that the use of NOACs seem
to be as effective and safe as warfarin/LMWH for the VTE
prophylaxis in cancer patients (Larsen et al., 2014; Van Der
Hulle et al., 2014; Vedovati et al., 2015; Brunetti et al., 2017; Di
Minno et al., 2017). However, insufficient sample size was the
main limition for aboved meta-analysis studies. Citation: Yan Y-D, Zhang C, Shen L, Su Y-J,
Liu X-Y, Wang L-W and Gu Z-C (2018)
Net Clinical Benefit of Non-vitamin K
Antagonist Oral Anticoagulants for
Venous Thromboembolism
Prophylaxis in Patients With Cancer: A
Systematic Review and Trade-Off
Analysis From 9 Randomized
Controlled Trials. Front. Pharmacol. 9:575. doi: 10.3389/fphar.2018.00575 June 2018 | Volume 9 | Article 575 Frontiers in Pharmacology | www.frontiersin.org 1 Yan et al. NCB of NOACs in Cancer Patients for broad NCB) and no-cancer patients (OR: 0.75, 95%CI: 0.54-0.96 for narrow NCB;
OR: 0.85, 95%CI: 0.67–1.04 for broad NCB), with the estimates mainly from VTE
patients receiving long-term warfarin treatment. In conclusion, NOACs may represent
a better NCB property compared to traditional anticoagulants in cancer patients who
need long-term anticoagulation treatment. Keywords: non-vitamin K antagonist oral anticoagulants, dabigatran, rivaroxaban, apixaban, edoxaban, cancer,
venous thromboembolism, net clinical benefit Keywords: non-vitamin K antagonist oral anticoagulants, dabigatran, rivaroxaban, apixaban, edoxaban, cancer,
venous thromboembolism, net clinical benefit INTRODUCTION symptomatic or incidental VTE have been published in Feb 2018,
which met the sample size requirement for estimating a reduction
in VTE from 3 to 5% (Raskob et al., 2018). In addition, the
concept of net clinical benefit (NCB) has been growingly used to
quantify both thromboembolism and hemorrhage in the field of
anticoagulant treatment, while no NCB studies to date have been
specifically addressed in NOACs-treated patients with cancer. Hence, it is necessary to address this important knowledge gap by
using a trade-offanalysis from previous randomized controlled
trials (RCTs) as well as recently published Hokusai-Cancer trial. Venous thromboembolism (VTE) is highly prevalent in patients
with cancer, occurring up to 15% of cancer patients during
the course of their diseases (Caine et al., 2002; Elalamy et al.,
2017). A prominent role is attributed to ability of tumor
cells to destabilize the coagulation system including releasing
procoagulant proteases, expressing tissue factor on cancer cells,
deriving microvesicles, as well as altering the extracellular
matrix of the cancer cell milieu (Nickel et al., 2016). These
patients, when receiving anticoagulant treatment, have a high
risk of recurrent VTE, while prossessing the danger of bleeding
complications (Prandoni et al., 2002). Therefore, the optimal
anticoagulant strategy for banlancing VTE and bleeding poses
a major challenge in this fragile population. Lee AYY et al
conducted the CLOT trial comparing dalteparin with warfarin
for preventing recurrent VTE in 672 patients with cancer, and
revealed that low-molecular-weight heparin (LMWH) was more
effective than vitamin-K antagonists (VKAs) in reducing the risk
of recurrent VTE, without increasing the major bleeding risk (Lee
et al., 2003). LMWH thus is the recommended anticoagulant for
the treatment of cancer-associated VTE. Data Sources and Searches Encouragingly,
the latest Hokusai-Cancer trial involving large sample size of
1,050 patients with predominantly advanced cancer and acute Frontiers in Pharmacology | www.frontiersin.org Data Extraction and Quality Evaluation All data that followed intention-to-treat principle were extracted
independently by two reviewers (Yi-Dan Yan and Chi Zhang)
using a priori designed form that included number of patients,
mean age, sex, mean weight, body mass index (BMI), type
of VTE, duration of follow-up, VTE events, MBEs, CRBEs in
patients with or without cancer. The methodological quality
of trials was evaluated based on the Cochrane Collaboration
Risk of Bias Tool, which included random sequence generation,
allocation concealment, blinding, incomplete outcome data,
selective reporting, and other bias (Higgins et al., 2011; Wei
et al., 2016). Potential publication bias was evaluated by visually
inspecting funnel plots if more than 10 studies were included (Gu
et al., 2017). FIGURE 1 | Flow diagram for the selection of eligible randomized controlled
trials. NCB Analysis in Patients With and Without
Cancer Cancer
The NCB analyses in patients with cancer were presented in
Figures 2A,B. NOACs had a superior NCB than traditional
anticoagulation,
irrespective
of
narrow
NCB
(OR:
0.68,
95%CI: 0.50–0.85) and broad NCB (OR: 0.76, 95%CI: 0.61–
0.91). No significant heterogeneity was detected in both
narrow NCB analysis (I2: 0.0%; P = 0.792) and broad NCB
analysis (I2: 1.0%; P = 0.438). In patients without cancer,
as shown in Figures 3A,B, narrow NCB of NOACs was
superior
when
compared
with
traditional
anticoagulation
(OR: 0.75, 95%CI: 0.54–0.96), and broad NCB showed a
borderline
significant
result
with
NOACs
vs. traditional
anticoagulation (OR: 0.85, 95%CI: 0.67–1.04). The considerable Study Selection and Outcomes y
The primacy outcomes were narrow NCB weighting both VTE
events and major bleeding events (MBEs), as well as broad
NCB balancing VTE events and clinically relevant bleeding
events (CRBEs: MBEs and clinically relevant non-major bleeding
events). Studies were considered potentially eligible for this
systematic review if they met the following predetermined
criteria: (1) only RCTs that reported intested data of patients
with or without cancer were included; and (2) VTE events,
MBEs, or CRBEs were objectively assessed in NOACs groups and June 2018 | Volume 9 | Article 575 Frontiers in Pharmacology | www.frontiersin.org 2 Yan et al. NCB of NOACs in Cancer Patients traditional anticoagulation groups, respectively. For duplicate
publications of the same RCTs, the most relevent to our inclusion
criteria was considered. When regarding possibly selective bias,
two reviewers (Yi-Dan Yan and Chi Zhang) were blinded to
authors’ names, journal names, and publication years of the
papers, and all disagreements were resolved through discussion
and the opinion of a third reviewer (Zhi-Chun Gu). FIGURE 1 | Flow diagram for the selection of eligible randomized controlled
trials. Data Analysis y
Odds ratios (ORs) and their 95% confidence intervals (CIs) of
VTE events and bleeding events were calculated by employing
a random-effects model. The narrow NCB of NOACs vs. traditional anticoagulation was calculated by pooling ORs of
VTE and MBEs, with a weighting of 1.0. Similarly, the broad
NCB was conducted by merging ORs of VTE and CRBEs, with
a weighting of 1.0. The overall estimates were presented in
forest plots, and weighting of each study was assigned on the
basis of event rate and sample size. Heterogeneity, defined as
variation beyond chance, was evaluated through the I2 test and Q
statistic that measures the percentage of total variation between
studies. I2 of >50% indicated considerable heterogeneity, and
a p <0.05 at Q statistic represented a significant heterogeneity
(Higgins et al., 2003). Subgroup analyses of NCB were calculated
by different patients (VTE patients or acutely ill patients),
comparisons (vitamin-K antagonists or low-molecular-weight
heparin), and duration of follow-up (≤6 months or >6 months). Sensitivity analysis was also performed for detecting the effect
of a single trial by sequential elimination of each trial from the
pool, and afterward to reassess the overall effects. All statistical
analyses were performed by using STATA software (version13,
Statacorp, College Station, Texas, USA), and P < 0.05 indicated a
statistically significant difference. et al., 2018). The characteristics and defination of outcomes
in included RCTs were presented in Table 1 and Supplemental
Table 1. A total of 41,454 patients were enrolled, of which 2,902
were cancer patients (1,499 patients exposed to NOACs and
1,403 patients exposed to traditional anticoagulation) and 38,552
were no-cancer patients (19,213 patients allocated to NOACs and
19,339 patients allocated to traditional anticoagulation). Of these
9 studies, 7 studies concerned about patients with VTE and 2
studies concerned about acutely ill patients. The median age of
patients ranged from 54 to 71 years and the percentage of male
ranged from 40 to 60. Also, The duration of follow-up ranged
from 1 to 12 months across the 9 trials. All trials satisfied bias tool
items with the exception of EINSTEIN trial and Hokusai-Cancer
trial, which were open-label studies. Thus, the included studies
had low bias overall, meaning that the quality of the included
trials was very high (Table 2). Frontiers in Pharmacology | www.frontiersin.org Study Evaluation The flow diagram for study selection was shown in Figure 1. The literature search yielded 4,228 records, of which 47 full-
text articles were obtained to further assess for eligibility, and 9
eligible RCTs were included in the final analyses (Schulman et al.,
2009, 2014; Bauersachs et al., 2010; Goldhaber et al., 2011; Büller
et al., 2012, 2013; Agnelli et al., 2013; Cohen et al., 2013; Raskob June 2018 | Volume 9 | Article 575 Frontiers in Pharmacology | www.frontiersin.org 3 NCB of NOACs in Cancer Patients Yan et al. Yan et al. Yan et al. NCB of NOACs in Cancer Patients
TABLE 1 | Summarized characteristics of included randomized controlled trials. Source
Patients
Drugs
N
Mean age (y)
Male
(%)
Mean weight
BMI
(kg/m2)
CCr30-50
(%)
DVT
(%)
Follow
up(month)
AMPLIFY
Cancer
Apixaban
88
65.5
56.8
79.16
NA
8
67
6
Enoxaparin/warfarin
81
65.1
60.5
81.69
NA
13.6
88
No Cancer
Apixaban
2417
56
59.1
84.88
NA
5.2
66
Enoxaparin/warfarin
2444
55.6
59.5
84.84
NA
4.7
69
EINSTEIN-DVT/PE
Cancer
Rivaroxaban
258
NA
59
NA
27.4
13*
NA
12
Enoxaparin/warfarin
204
NA
53
NA
26.7
17*
NA
No Cancer
Rivaroxaban
3563
NA
55
NA
28.2
7*
NA
Enoxaparin/warfarin
3594
NA
54
NA
28.2
7*
NA
Hokusai
Cancer
Edoxaban
109
67
50
NA
NA
NA
58
12
Warfarin
99
66
61
NA
NA
NA
59
RE-COVER-I/II
Cancer
Dabigatran
114
63.5
49
78.1
27.6
NA
75
6
Warfarin
107
65.3
45
76.1
26.8
NA
74
No Cancer
Dabigatran
2380
54.3
40.1
84.7
28.7
NA
69
Warfarin
2392
54
40.6
84
28.5
NA
68
Hokusai-Cancer
Cancer
Edoxaban
522
64.3
53.1
78.8
NA
7.3
37
12
LMWH
524
63.7
50.2
79.1
NA
6.5
37
MAGELLAN
All
Rivaroxaban
4050
71
55.6
77.5
28.2
21.5
NA
1
LMWH
4050
71
52.7
77.3
28.2
21.5
NA
ADOPT
All
Apixaban
3255
66.8
50
NA
NA
NA
NA
1
LMWH
3273
66.7
48.2
NA
NA
NA
NA
BMI, body mass index; CCr, creatinine clearance rate; DVT, deep vein thrombosis; NA, not available; N, number of patients; *EINSTEIN-DVT/PE study provided combined data of
patients whose CCr < 50 ml/min. TABLE 2 | Quality assessment. Study Evaluation Source
Patients
Drugs
N
Mean age (y)
Male
(%)
Mean weight
BMI
(kg/m2)
CCr30-50
(%)
DVT
(%)
Follow
up(month)
AMPLIFY
Cancer
Apixaban
88
65.5
56.8
79.16
NA
8
67
6
Enoxaparin/warfarin
81
65.1
60.5
81.69
NA
13.6
88
No Cancer
Apixaban
2417
56
59.1
84.88
NA
5.2
66
Enoxaparin/warfarin
2444
55.6
59.5
84.84
NA
4.7
69
EINSTEIN-DVT/PE
Cancer
Rivaroxaban
258
NA
59
NA
27.4
13*
NA
12
Enoxaparin/warfarin
204
NA
53
NA
26.7
17*
NA
No Cancer
Rivaroxaban
3563
NA
55
NA
28.2
7*
NA
Enoxaparin/warfarin
3594
NA
54
NA
28.2
7*
NA
Hokusai
Cancer
Edoxaban
109
67
50
NA
NA
NA
58
12
Warfarin
99
66
61
NA
NA
NA
59
RE-COVER-I/II
Cancer
Dabigatran
114
63.5
49
78.1
27.6
NA
75
6
Warfarin
107
65.3
45
76.1
26.8
NA
74
No Cancer
Dabigatran
2380
54.3
40.1
84.7
28.7
NA
69
Warfarin
2392
54
40.6
84
28.5
NA
68
Hokusai-Cancer
Cancer
Edoxaban
522
64.3
53.1
78.8
NA
7.3
37
12
LMWH
524
63.7
50.2
79.1
NA
6.5
37
MAGELLAN
All
Rivaroxaban
4050
71
55.6
77.5
28.2
21.5
NA
1
LMWH
4050
71
52.7
77.3
28.2
21.5
NA
ADOPT
All
Apixaban
3255
66.8
50
NA
NA
NA
NA
1
LMWH
3273
66.7
48.2
NA
NA
NA
NA
BMI, body mass index; CCr, creatinine clearance rate; DVT, deep vein thrombosis; NA, not available; N, number of patients; *EINSTEIN-DVT/PE study provided combined data of
patients whose CCr < 50 ml/min. TABLE 1 | Summarized characteristics of included randomized controlled trials. BMI, body mass index; CCr, creatinine clearance rate; DVT, deep vein thrombosis; NA, not available; N, number of patients; *EINSTEIN-DVT/PE study provided combined data of
patients whose CCr < 50 ml/min. TABLE 2 | Quality assessment. Study
Random sequence
generation
Allocation
concealment
Blinding of
participants and
personnel
Blinding of outcome
assessment
Incomplete
outcome data
Selective
reporting
Other
bias
AMPLIFY, 2013
L
L
L
L
L
L
L
EINSTEIN-DVT, 2010
L
U
H
L
L
L
L
EINSTEIN-PE, 2012
L
U
H
L
L
L
L
Hokusai-VTE, 2013
L
U
L
L
L
L
L
RE-COVER I, 2009
L
L
L
L
L
L
L
RE-COVER II, 2014
L
U
L
L
L
L
L
Hokusai-Cancer, 2017
L
U
H
L
L
L
L
MAGELLAN, 2013
L
U
L
L
L
L
L
ADOPT, 2011
L
U
L
L
L
L
L
L, low risk; U, unclear risk; H, high risk. Study Evaluation obtained in acutely ill patients due to limited sample size
(OR: 1.39, 95%CI: 0.46–2.33 for broad NCB in cancer patients;
OR: 1.54, 95%CI: 0.17–3.25 for broad NCB in no-cancer
patients). In terms of different comparators, NOACs provided
a better NCB compared with warfarin, regardless of patients
with cancer (OR: 0.79, 95%CI: 0.61–0.98 for narrow NCB; OR:
0.71, 95%CI: 0.47–0.94 for broad NCB) and without cancer
(OR: 0.79, 95%CI: 0.61–0.98 for broad NCB). However, these
positive NCBs were not observed in comparison to LMWH
(OR: 1.10, 95%CI: 0.03–2.17 for narrow NCB in cancer patients;
OR: 1.05, 95%CI: 0.55–1.55 for broad NCB in cancer patients; heterogeneity was observed in narrow NCB analysis (I2:
74.9%; P < 0.01) as well as broad NCB analysis (I2: 89.3%;
P<0.01). Study Evaluation Study
Random sequence
generation
Allocation
concealment
Blinding of
participants and
personnel
Blinding of outcome
assessment
Incomplete
outcome data
Selective
reporting
Other
bias
AMPLIFY, 2013
L
L
L
L
L
L
L
EINSTEIN-DVT, 2010
L
U
H
L
L
L
L
EINSTEIN-PE, 2012
L
U
H
L
L
L
L
Hokusai-VTE, 2013
L
U
L
L
L
L
L
RE-COVER I, 2009
L
L
L
L
L
L
L
RE-COVER II, 2014
L
U
L
L
L
L
L
Hokusai-Cancer, 2017
L
U
H
L
L
L
L
MAGELLAN, 2013
L
U
L
L
L
L
L
ADOPT, 2011
L
U
L
L
L
L
L
L, low risk; U, unclear risk; H, high risk. heterogeneity was observed in narrow NCB analysis (I2:
74.9%; P < 0.01) as well as broad NCB analysis (I2: 89.3%;
P<0.01). NCB Analysis Based on Different Patients,
Comparators, and Follow-Up Duration
obtained in acutely ill patients due to limited sample size
(OR: 1.39, 95%CI: 0.46–2.33 for broad NCB in cancer patients;
OR: 1.54, 95%CI: 0.17–3.25 for broad NCB in no-cancer
patients). In terms of different comparators, NOACs provided
a better NCB compared with warfarin, regardless of patients
with cancer (OR: 0.79, 95%CI: 0.61–0.98 for narrow NCB; OR: TABLE 1 | Summarized characteristics of included randomized controlled trials. NCB Analysis Based on Different Patients,
Comparators, and Follow-Up Duration NCBs analyses for subgroups were summarized in Tables 3, 4. As for VTE patients, the use of NOACs showed a positive
broad NCB when compared to traditional anticoagulation in
patients with cancer (OR: 0.76, 95%CI: 0.58–0.93) and without
cancer (OR: 0.79, 95%CI: 0.61–0.98). No positive results were June 2018 | Volume 9 | Article 575 Frontiers in Pharmacology | www.frontiersin.org 4 NCB of NOACs in Cancer Patients Yan et al. Yan et al. yses of narrow net clinical benefit (A) and broad net clinical benefit (B) in patients with cancer. ES indicates Odds ratio; 95%CI indicates 95%
al. %CI
0 17 3 25 f
b
d NCB i
i
l i
(OR 0 68 95%CI 0 47 0 89) C
i
l row net clinical benefit (A) and broad net clinical benefit (B) in patients with cancer. ES indicates Odds ratio; 95%CI indicates 95% FIGURE 2 | Analyses of narrow net clinical benefit (A) and broad net clinical benefit (B) in patients with cancer. ES indicates Odds ratio; 95%CI indicates 95%
confidence interval. anticoagulation (OR: 0.68, 95%CI: 0.47–0.89). Consistent results
were observed in patients without cancer (OR: 0.81, 95%CI:
0.65–0.97 for narrow NCB; OR: 0.87, 95%CI: 0.79–0.95 for
broad NCB). OR: 1.54, 95%CI: 0.17–3.25 for broad NCB in no-cancer
patients). With regard to follow-up duration, the results over 6
months showed that cancer patients treated with NOACs were
associated with a better narrow NCB compared with traditional Frontiers in Pharmacology | www.frontiersin.org June 2018 | Volume 9 | Article 575 NCB of NOACs in Cancer Patients Yan et al. Yan et al. yses of narrow net clinical benefit (A) and broad net clinical benefit (B) in patients without cancer. ES indicates Odds ratio; 95%CI indicates 95%
al. FIGURE 3 | Analyses of narrow net clinical benefit (A) and broad net clinical benefit (B) in patients without cancer. ES indicates Odds ratio; 95%CI indicates 95%
confidence interval. FIGURE 3 | Analyses of narrow net clinical benefit (A) and broad net clinical benefit (B) in patients without cancer. ES indicates Odds ratio; 95%CI indicates 95%
fid
i
l FIGURE 3 | Analyses of narrow net clinical benefit (A) and broad net clinical benefit (B) in patients without cancer. ES indicates Odds ratio; 95%CI indicates 95%
confidence interval. Sensitivity Analyses and Publication Bias Sensitivity Analyses and Publication Bias Results of sensitivity analyses, including broad NCBs and narrow
NCBs, were consistent with those of the primacy analyses
(Table 5). Publication bias was not performed because of the
limited study number of 9. NCB that incorporates both the risk of VTE and major bleeding
provides a more quantitatively informed basis for the decision-
making on the optimal anticoagulant therapy in patients with June 2018 | Volume 9 | Article 575 Frontiers in Pharmacology | www.frontiersin.org NCB of NOACs in Cancer Patients Yan et al. TABLE 4 | Subgroup analyses for NCB in patients without cancer. Outcomes
No. of
studies
OR
95%CI
Homogeneity
I2 (%)
p-value
NARROW NCB IN NO CANCER
Follow up
≤6 months
3
0.72
0.32–1.12
83.00
0.00
>6 months
3
0.81
0.65–0.97
23.80
0.27
BROAD NCB IN NO CANCER
Patients
VTE patients
6
0.79
0.61–0.98
88.80
0.0
Acutely ill patients
1
1.54
0.17–3.25
95.00
0.00
CLASS OF CONTROL DRUGS
Warfarin
6
0.79
0.61–0.98
88.80
0.00
LMWH
1
1.54
0.17–3.25
95.00
0.00
Follow up
≤6 months
4
0.85
0.58–1.12
89.40
0.00
> 6 months
3
0.87
0.79–0.95
0.00
0.69
NCB, net clinical benefit; LMWH, low molecular weight heparin; VTE, venous
thromboembolism; OR, odds ratios; CI, confidence intervals. TABLE 3 | Subgroup analyses for NCB in patients with cancer. Outcomes
No. of
studies
OR
95%CI
Homogeneity
I2 (%)
p-value
NARROW NCB IN CANCER
Class of control drugs
Warfarin
6
0.61
0.28–0.94
0.00
1.00
LMWH
1
1.10
0.03–2.17
76.9
0.04
Follow up
≤6 months
3
0.68
0.19–1.18
0.00
0.96
>6 months
4
0.68
0.47–0.89
3.40
0.40
BROAD NCB IN CANCER
Patients
VTE patients
7
0.76
0.58–0.93
11.20
0.34
Acutely ill patients
2
1.39
0.46–2.33
0.00
0.77
CLASS OF CONTROL DRUGS
Warfarin
6
0.71
0.49–0.90
0.00
0.82
LMWH
3
1.05
0.55–1.55
45.8
0.10
Follow up
≤6 months
5
0.75
0.44–1.07
0.00
0.65
>6 months
4
0.82
0.55–1.08
37.80
0.15
NCB, net clinical benefit; LMWH, low molecular weight heparin; VTE, venous
thromboembolism; OR, odds ratios; CI, confidence intervals. TABLE 4 | Subgroup analyses for NCB in patients without cancer. TABLE 3 | Subgroup analyses for NCB in patients with cancer. NARROW NCB IN NO CANCER NCB, net clinical benefit; LMWH, low molecular weight heparin; VTE, venous
thromboembolism; OR, odds ratios; CI, confidence intervals. et al., 2013). Sensitivity Analyses and Publication Bias The use of anticoagulants is the standard treatment
for the prevention of VTE in cancer patients, while an 8–
10% annual bleeding events occurs during anticoagulation
therapy (Brose and Lee, 2008). Hence, it is essential to balance
the benefit and risk of anticoagulation therapy in this fragile
population. cancer. To the best of our knowledge, this is the largest sample
size analysis to pool 2,902 cancer patients for evaluation of
NCB of NOACs. Our results indicated that NCB of NOACs,
in both cancer and no cancer patients, were superior to that of
traditional anticoagulation, with the estimates mainly from VTE
patients, warfarin-treated patients, and patients with long-term
anticoagulation treatment (>6 months). p p
Prior meta-analysis studies that involved about 1,000 patients
have addressed that NOACs seem to be as effective and safe
as VKAs for the prevention of VTE in patients with cancer
(Larsen et al., 2014; Van Der Hulle et al., 2014; Vedovati
et al., 2015). In fact, at least 1500 patients should be analyzed
in order to demonstrate a reduction in VTE from 3 to 5%. Afterward, Brunetti et al reported a consistent result even after
pooling data with VKA and LMWH (Brunetti et al., 2017). The latest study by Di Minno et al, which separated data on
patients with active cancer and cancer history, suggested a
significantly lower risk of VTE and a non-significantly lower risk
of major bleeding for the use of NOACs in patients with active
cancer when compared to the use of VKAs (Di Minno et al.,
2017). However, direct head-to-head comparison with LMWH
is necessary before NOACs can be routinely appiled for cancer-
associated VTE patients. Encouragingly, the latest Hokusai-
Cancer trial, which included 1050 patients with predominantly
advanced cancer and acute symptomatic or incidental VTE,
showed that the use of edoxaban (Xa factor inhibitor) for
up to 12 months was non-inferior to the use of LMWH in
terms of the composite outcome of VTE or major bleeding
(Raskob et al., 2018). When regarding efficacy, the rate of
VTE was numerically lower with edoxaban than LMWH owing
to the lower rate recurrent symptomatic deep-vein thrombus
with edoxaban. While in terms of safety, the risk of major
bleeding was significantly increased with edoxaban than with
LMWH, which was mainly due to the higher risk of upper Frontiers in Pharmacology | www.frontiersin.org LIMITATIONS Several limitations need to be considered. Firstly, included
RCTs were not especially designed to assess VTE and bleeding
risk of NOACs in patients with cancer, with the expection
of Hokusai-Cancer trial. Therefore, the difference in the
baseline characteristics in patients allocated to NOACs and
VKAs/LMWH could not be excluded. Secondly, the definition
of outcomes was not same across included RCTs. However,
as revealed by the values of I2, heterogeneity among the
RCTs in patients with cancer was low in our random-effects
model. Thirdly, no admitted methods are available to estimate
the NCB in the field of VTE recently, thus we carried
out a trade-offanalysis between VTE and bleeding using a
weighting of 1.0. Fourthly, individual NOACs analysis was not
performed due to the limited studies. Fifthly, we have not
get access to patient-level data in relation to the type, the
stage or the location of cancer, making powerful subgroup
analysis unavailable. Sixth, included studies have not addressed
the impact of NOACs on different stakeholders including
healthcare providers, users, and policymakers. Finally, drug-
drug interaction information that impacts NOACs or VKAs
were not addressed in included studies, which may lead certain
bias. Accordingly, Further real-world studies are necessary to
be conducted for the assessment of NOACs use in these special
patients. OR, odds ratio; 95%CI, 95% confidence interval. gastrointestinal bleeding with edoxaban (Raskob et al., 2018). Accordingly, treatment with NOACs may reduce the risk of VTE
at the expense of increased risk of major bleeding. In the present
study, we focused on a core issue, namely, net clinical benefit
that weighted the benefit and risk, and analyzed it by pooling
currently avaiable RCTs. The results found that the use of NOACs
posed a better NCB than traditional anticoagulants in patients
with cancer. After further analysis based on subgroups, we recognized that
the positive results of NCB was derived mainly from trials on
NOACs vs. warfarin. For patients with warfarin therapy, time in
therapeutic range (TTR) could reflect the anticoagulation effects. During the management of RCTs, it is recognized that TTR would
be well controlled. Nevertheless, due to the frequent interactions
with chemothrapeutic agents and immunosuppressive agents in
anticancer therapy, TTR is hard to do well in practice. Compared
to LMWH, NOACs showed non-inferior NCB in patients with
cancer. The main contribution of the result came from recently
published Hokusai-Cancer trial (Raskob et al., 2018). LIMITATIONS In addition,
positive NCB with NOACs therapy in cancer patients was
primarily derived from VTE patients on long-term treatment (>6
months). Patients with VTE are usually recommended to receive
long-term anticoagulation therapy (Kearon et al., 2016), and the
superior NCB of NOACs emerges during the course of treatment. As for acutely ill patients, sample size limited our ability to make NCB in Patients With or Without Cancer At present, NOACs have been shown to be as effective as and safer
than traditional anticoagulation for VTE prevention in no-cancer
patients. Thus, the use of NOACs are currently recommended for
the treatment and prophylaxis of VTE by international clinical
guideline (Kearon et al., 2016). Our trade-offanalyses from
pooling 9 RCTs showed that no-cancer patients treated with
NOACs had a better NCB compared with patients treated with
traditional anticoagulation. Cancer has been associated with an increased risk of
thromboembolic events, and the coexist of cancer and VTE
confers significantly greater risk of this issue (Caine et al.,
2002; Prandoni et al., 2002). In a pooled analysis of data
from 38 study populations, the authors estimated the annual
incidence rate of VTE to be between 0.5 and 20% depending
on the cancer type, and found that cancers of the brain and
pancreas were associated with the highest risk of VTE (Horsted
et al., 2012). In a study from the USA, cancer patients with
VTE were three times more likely to be hospitalized, with an
additional seven hospital days relative to patients without VTE. This translated into significantly higher total healthcare costs
(US$74,959 vs. US$41,691 per patient; p < 0.001) (Khorana June 2018 | Volume 9 | Article 575 June 2018 | Volume 9 | Article 575 7 NCB of NOACs in Cancer Patients Yan et al. TABLE 5 | Sensitivity analyses. Study omitted
OR
95%CI
NARROW NCB IN CANCER
AMPLIFY
0.68
0.51-0.86
EINSTEIN-PE/DVT
0.69
0.51-0.87
Hokusai
0.68
0.51–0.86
RECOVER-I/II
0.67
0.49–0.85
Hokusai-Cancer
0.61
0.28–0.94
BROAD NCB IN CANCER
AMPLIFY
0.81
0.64–0.99
EINSTEIN-PE/DVT
0.76
0.58–0.94
Hokusai
0.82
0.62–1.02
RECOVER-I/II
0.77
0.58–0.95
Hokusai-Cancer
0.75
0.52–0.97
MAGELLAN
0.74
0.59–0.88
ADOPT
0.79
0.61–0.98
NARROW NCB IN NO CANCER
AMPLIFY
0.80
0.64–0.97
EINSTEIN-PE/DVT
0.76
0.48–1.04
Hokusai
0.72
0.46–0.99
RECOVER-I/II
0.74
0.50–0.98
BROAD NCB IN NO CANCER
AMPLIFY
0.89
0.71–1.08
EINSTEIN-PE/DVT
0.84
0.63–1.05
Hokusai
0.87
0.64–1.10
RECOVER-I/II
0.89
0.67–1.10
MAGELLAN
0.79
0.61–0.98
OR, odds ratio; 95%CI, 95% confidence interval. a definitive conclusions and further assessment of NCB in these
patients should be conducted. Clinical Challenge and Implication g
Treatment
and
prophylaxis
of
cancer-associated
VTE
is
challenging, and guidelines recommend treatment with LMWH
due to lower rate of thrombosis and similar rate of bleeding
when compared with VKAs (Farge et al., 2013; Lyman et al.,
2015; Kearon et al., 2016). However, the use of LMWH is
burdensome because it requires daily subcutaneous injecions,
which may limit its long-term adoption. Given this clincial
setting, NOACs may represent an alternative choice because
of oral route of administration, predictable dose response,
no need for laboratory monitoring, and greater flexibility
during
invasive
procedures. All
these
adventages
makes
NOACs more appealing for long-term prevention of VTE. Undeniably, interactions between chemothrapeutic agents and
immunosuppressive agents with NOACs are still possible, and
drugs that interfere P-glycoprotein or CYP3A4 may impact
the anticoagulation effect of NOACs (Voukalis et al., 2016). Thus, It is important to acknowledge that our results cannot
be extrapolated to the cancer patients with the presence of
potiental drug-drug interaction, but may only be applied to
patients with similar characteristics included in the present
analysis. Frontiers in Pharmacology | www.frontiersin.org REFERENCES (2012). Oral rivaroxaban for the treatment of symptomatic pulmonary
embolism. N. Engl. J. Med. 366, 1287–1297. doi: 10.1056/NEJMoa1113572 j
p
Lee, A. Y., Levine, M. N., Baker, R. I., Bowden, C., Kakkar, A. K., Prins, M., et al. (2003). Low-molecular-weight heparin versus a coumarin for the prevention of
recurrent venous thromboembolism in patients with cancer. N. Engl. J. Med. 349, 146–153. doi: 10.1056/NEJMoa025313 Caine, G. J., Stonelake, P. S., Lip, G. Y., and Kehoe, S. T. (2002). The
hypercoagulable state of malignancy: pathogenesis and current debate. Neoplasia 4, 465–473. doi: 10.1038/sj.neo.7900263 Lyman, G. H., Bohlke, K., and Falanga, A. (2015). Venous thromboembolism
prophylaxis and treatment in patients with cancer: American Society of Clinical
Oncology clinical practice guideline update. J. Oncol. Pract. 11, e442–e444. doi: 10.1200/JOP.2015.004473 Cohen, A. T., Spiro, T. E., Buller, H. R., Haskell, L., Hu, D., Hull, R., et al. (2013). Rivaroxaban for thromboprophylaxis in acutely ill medical patients. N. Engl. J. Med. 368, 513–523. doi: 10.1056/NEJMoa1111096 Di Minno, M. N. D., Ageno, W., Lupoli, R., Conte, G., Van Es, N., Buller, H. R., et al. (2017). Direct oral anticoagulants for the treatment of acute venous
thromboembolism in patients with cancer: a meta-analysis of randomised
controlled trials. Eur. Respir. J. 50:1701097. doi: 10.1183/13993003.01097-2017 Nickel, K. F., Labberton, L., Long, A. T., Langer, F., Fuchs, T. A., Stavrou,
E. X., et al. (2016). The polyphosphate/factor XII pathway in cancer-
associated
thrombosis:
novel
perspectives
for
safe
anticoagulation
in
patients
with
malignancies. Thromb
Res. 141
(Suppl. 2),
S4–S7. doi: 10.1016/S0049-3848(16)30353-X Elalamy, I., Mahe, I., Ageno, W., and Meyer, G. (2017). Long-term treatment
of cancer-associated thrombosis: the choice of the optimal anticoagulant. J. Thromb. Haemost. 15, 848–857. doi: 10.1111/jth.13659 Prandoni, P., Lensing, A. W., Piccioli, A., Bernardi, E., Simioni, P., Girolami, B.,
et al. (2002). Recurrent venous thromboembolism and bleeding complications
during anticoagulant treatment in patients with cancer and venous thrombosis. Blood 100, 3484–3488. doi: 10.1182/blood-2002-01-0108 Farge, D., Debourdeau, P., Beckers, M., Baglin, C., Bauersachs, R. M., Brenner, B.,
et al. (2013). International clinical practice guidelines for the treatment and
prophylaxis of venous thromboembolism in patients with cancer. J. Thromb. Haemost. 11, 56–70. doi: 10.1111/jth.12070 Raskob, G. E., Van Es, N., Verhamme, P., Carrier, M., Di Nisio, M.,
Garcia,
D.,
et
al. (2018). Edoxaban
for
the
treatment
of
cancer-
associated venous thromboembolism. N. Engl. J. Med. 378, 615–624. doi: 10.1056/NEJMoa1711948 Goldhaber, S. Z., Leizorovicz, A., Kakkar, A. K., Haas, S. REFERENCES risk of bias in randomised trials. BMJ
343:d5928. doi: 10.1136/bmj. d5928 Agnelli, G., Buller, H. R., Cohen, A., Curto, M., Gallus, A. S., Johnson, M., et al. (2013). Oral apixaban for the treatment of acute venous thromboembolism. N. Engl. J. Med. 369, 799–808. doi: 10.1056/NEJMoa1302507 Higgins,
J. P.,
Thompson,
S. G.,
Deeks,
J. J.,
and
Altman,
D. G. (2003). Measuring inconsistency in meta-analyses. BMJ 327, 557–560. doi: 10.1136/bmj.327.7414.557 g
Bauersachs, R., Berkowitz, S. D., Brenner, B., Buller, H. R., Decousus, H., Gallus, A. S., et al. (2010). Oral rivaroxaban for symptomatic venous thromboembolism. N. Engl. J. Med. 363, 2499–2510. doi: 10.1056/NEJMoa1007903 Horsted, F., West, J., and Grainge, M. J. (2012). Risk of venous thromboembolism
in patients with cancer: a systematic review and meta-analysis. PLoS Med. 9:e1001275. doi: 10.1371/journal.pmed.1001275 N. Engl. J. Med. 363, 2499–2510. doi: 10.1056/NEJMoa1007903 Brose, K. M., and Lee, A. Y. (2008). Cancer-associated thrombosis: prevention and
treatment. Curr. Oncol. 15, S58–S67. doi: 10.3747/co.2008.177 Brose, K. M., and Lee, A. Y. (2008). Cancer-associated thrombosis: Kearon, C., Akl, E. A., Ornelas, J., Blaivas, A., Jimenez, D., Bounameaux,
H., et al. (2016). Antithrombotic therapy for VTE disease: chest guideline
and expert panel report. Chest 149, 315–352. doi: 10.1016/j.chest.2015. 11.026 treatment. Curr. Oncol. 15, S58–S67. doi: 10.3747/co.2008.177 Brunetti, N. D., Gesuete, E., De Gennaro, L., Correale, M., Caldarola, P.,
Gaglione, A., et al. (2017). Direct oral anti-coagulants compared with vitamin-
K inhibitors and low-molecular-weight-heparin for the prevention of venous
thromboembolism in patients with cancer: a meta-analysis study. Int. J. Cardiol. 230, 214–221. doi: 10.1016/j.ijcard.2016.12.168 Khorana, A. A., Dalal, M. R., Lin, J., and Connolly, G. C. (2013). Health care costs
associated with venous thromboembolism in selected high-risk ambulatory
patients with solid tumors undergoing chemotherapy in the United States. Clinicoecon. Outcomes Res. 5, 101–108. doi: 10.2147/CEOR.S39964 Clinicoecon. Outcomes Res. 5, 101–108. doi: 10.2147/CEOR.S3996 Büller, H. R., Decousus, H., Grosso, M. A., Mercuri, M., Middeldorp, S.,
Prins, M. H., et al. (2013). Edoxaban versus warfarin for the treatment of
symptomatic venous thromboembolism. N. Engl. J. Med. 369, 1406–1415. doi: 10.1056/NEJMoa1306638 Larsen, T. B., Nielsen, P. B., Skjoth, F., Rasmussen, L. H., and Lip, G. Y. (2014). Non-vitamin K antagonist oral anticoagulants and the treatment
of venous thromboembolism in cancer patients: a semi systematic review
and meta-analysis of safety and efficacy outcomes. PLoS ONE 9:e114445. doi: 10.1371/journal.pone.0114445 Büller, H. R., Prins, M. H., Lensin, A. W., Decousus, H., Jacobson, B. F., Minar,
E., et al. SUPPLEMENTARY MATERIAL The Supplementary Material for this article can be found
online
at:
https://www.frontiersin.org/articles/10.3389/fphar. 2018.00575/full#supplementary-material AUTHOR CONTRIBUTIONS Z-CG
and
L-WW
are
the
guarantors
of
the
entire
manuscript. Y-DY
and
CZ
contributed
to
the
study
conception
and
design;
data
acquisition,
analysis,
and
interpretation; drafting of the manuscript; critical revision
of the manuscript for important intellectual content; and
final approval of the version to be published. LS, Y-JS, and
X-YL
contributed
to
the
data
acquisition,
analysis,
and
interpretation. FUNDING cancer. Hence, NOACs may represent an alternative choice
in cancer patients who are expected long-term treatment of
anticoagulation. This work was supported by the Science Fund of Hospital
Pharmacy of Shanghai Jiaotong University School of Medicine
(JDYX2016ZD003), Program for Key Discipline of Clinical
Pharmacy of Shanghai (2016-40044-002), and Program for Key
but Weak Discipline of Shanghai Municipal Commission of
Health and Family Planning (2016ZB0304). This work was supported by the Science Fund of Hospital
Pharmacy of Shanghai Jiaotong University School of Medicine
(JDYX2016ZD003), Program for Key Discipline of Clinical
Pharmacy of Shanghai (2016-40044-002), and Program for Key
but Weak Discipline of Shanghai Municipal Commission of
Health and Family Planning (2016ZB0304). CONCLUSIONS The use of NOACs, compared with traditional anticoagulation,
conferred a better net clinical benefit profile in patients with Frontiers in Pharmacology | www.frontiersin.org June 2018 | Volume 9 | Article 575 8 NCB of NOACs in Cancer Patients Yan et al. REFERENCES K., Merli, G., Knabb, R. M.,
et al. (2011). Apixaban versus enoxaparin for thromboprophylaxis in medically
ill patients. N. Engl. J. Med. 365, 2167–2177. doi: 10.1056/NEJMoa1110899 Schulman, S., Kakkar, A. K., Goldhaber, S. Z., Schellong, S., Eriksson, H.,
Mismetti, P., et al. (2014). Treatment of acute venous thromboembolism
with dabigatran or warfarin and pooled analysis. Circulation 129, 764–772. doi: 10.1161/CIRCULATIONAHA.113.004450 Gu, Z., Zhang, C., Wei, A., Cui, M., Pu, J., Lin, H., et al. (2017). Incidence
and risk of respiratory tract infection associated with specific drug therapy
in pulmonary arterial hypertension: a systematic review. Sci. Rep. 7:16218. doi: 10.1038/s41598-017-16349-7 Schulman,
S.,
Kearon,
C.,
Kakkar,
A. K.,
Mismetti,
P.,
Schellong,
S.,
Eriksson, H., et al. (2009). Dabigatran versus warfarin in the treatment Higgins, J. P., Altman, D. G., Gotzsche, P. C., Juni, P., Moher, D., Oxman,
A. D., et al. (2011). The Cochrane Collaboration’s tool for assessing June 2018 | Volume 9 | Article 575 Frontiers in Pharmacology | www.frontiersin.org 9 Yan et al. NCB of NOACs in Cancer Patients of acute venous thromboembolism. N. Engl. J. Med. 361, 2342–2352. doi: 10.1056/NEJMoa0906598 of acute venous thromboembolism. N. Engl. J. Med. 361, 2342–2352. doi: 10.1056/NEJMoa0906598 Wei, A., Gu, Z., Li, J., Liu, X., Wu, X., Han, Y., et al. (2016). Clinical
adverse effects of endothelin receptor antagonists: insights from the meta-
analysis of 4894 patients from 24 randomized double-blind placebo-
controlled clinical trials. J. Am Heart Assoc. 5:e003896. doi: 10.1161/JAHA.116. 003896 Schulman,
S.,
Kearon,
C.,
Kakkar,
A. K.,
Schellong,
S.,
Eriksson,
H.,
Baanstra, D., et al. (2013). Extended use of dabigatran, warfarin, or
placebo in venous thromboembolism. N. Engl. J. Med. 368, 709–718. doi: 10.1056/NEJMoa1113697 Conflict of Interest Statement: The authors declare that the research was
conducted in the absence of any commercial or financial relationships that could
be construed as a potential conflict of interest. Van Der Hulle, T., Den Exter, P. L., Kooiman, J., Van Der Hoeven, J. J., Huisman,
M. V., and Klok, F. A. (2014). Meta-analysis of the efficacy and safety
of new oral anticoagulants in patients with cancer-associated acute venous
thromboembolism. J. Thromb. Haemost. 12, 1116–1120. doi: 10.1111/jth. 12605 Copyright © 2018 Yan, Zhang, Shen, Su, Liu, Wang and Gu. This is an open-access
article distributed under the terms of the Creative Commons Attribution License (CC
BY). Frontiers in Pharmacology | www.frontiersin.org June 2018 | Volume 9 | Article 575 REFERENCES The use, distribution or reproduction in other forums is permitted, provided
the original author(s) and the copyright owner are credited and that the original
publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these
terms. Vedovati, M. C., Germini, F., Agnelli, G., and Becattini, C. (2015). Direct oral
anticoagulants in patients with VTE and cancer: a systematic review and
meta-analysis. Chest 147, 475–483. doi: 10.1378/chest.14-0402 Voukalis, C., Lip, G. Y., and Shantsila, E. (2016). Drug-drug interactions of non-
vitamin K oral anticoagulants. Exp. Opin. Drug Metab. Toxicol. 12, 1445–1461. doi: 10.1080/17425255.2016.1225037 June 2018 | Volume 9 | Article 575 Frontiers in Pharmacology | www.frontiersin.org 10
|
https://openalex.org/W2175154879
|
https://link.springer.com/content/pdf/10.1007/s40846-015-0087-7.pdf
|
English
| null |
Combination Therapy Using Chelating Agent and Zinc for Wilson’s Disease
|
Journal of medical and biological engineering
| 2,015
|
cc-by
| 8,134
|
Combination Therapy Using Chelating Agent and Zinc
for Wilson’s Disease Jui-Chi Chen1 • Cheng-Hung Chuang1,2 • Jing-Doo Wang1,3 • Chi-Wei Wang4 Jui-Chi Chen1 • Cheng-Hung Chuang1,2 • Jing-Doo Wang1,3 • Chi-Wei Wang4 Received: 26 January 2015 / Accepted: 15 June 2015 / Published online: 19 November 2015
The Author(s) 2015. This article is published with open access at Springerlink.com Abstract
There
is
no
clear
international
consensus
regarding the optimal medication therapy for treating
Wilson’s disease (WD). This study systematically reviews
the effectiveness of various medication therapies in com-
mon use, specifically focusing on preliminary findings
concerning the combination of a chelating agent and zinc. A systematic PubMed search was executed to locate orig-
inal studies on the effectiveness of commonly used medi-
cations for WD published between January 1989 and
August 2014. The results were used to conduct a systematic
review of studies on combination therapies. A total of 17
combination therapy studies involving 1056 patients were
reviewed. These were analyzed in terms of data on effec-
tiveness, adverse effects, and mortality. Results from a
pooled analysis indicate that combination therapies for
hepatic patients were significantly less effective than the
same therapies for neurological manifestations (47.1 vs. 78.6 %; pooled relative risk ratio (RR): 0.63, 95 % confi-
dence interval CI 0.43–0.94; p = 0.02). Data from a sub-
group analysis show that the combination therapy of
penicillamine plus zinc sulfate resulted in a significantly
higher mortality rate compared to all other combination
therapy types (16.3 vs. 4.7 %; RR: 3.51, 95 % CI 1.54–8.00; p \ 0.001). The use of combination therapies involving zinc
and a chelator should be carefully monitored with close
clinical observations and frequent biochemical tests, espe-
cially for WD patients with hepatic manifestations. Keywords
Biomedical informatics Wilson’s disease
Combination therapy Effectiveness Safety J. Med. Biol. Eng. (2015) 35:697–708
DOI 10.1007/s40846-015-0087-7 REVIEW ARTICLE & Jui-Chi Chen
rikki@asia.edu.tw
1
Department of Computer Science and Information
Engineering, Asia University, Taichung 41354, Taiwan
2
Department of Medical Research, China Medical University
Hospital, Taichung 40402, Taiwan
3
Department of Biomedical Informatics, Asia University,
Taichung 41354, Taiwan
4
Department of Internal Medicine, Ben Tang Cheng Ching
Hospital, Taichung 41364, Taiwan 2.1 Search Strategy and Study Selection We performed a systematic search of the National Center
for Biotechnology Information’s PubMed database [35] for
original WD treatment studies published between January
1989 and August 2014. Search keywords were ‘‘Wilson’s
disease’’ or its synonyms (including ‘‘HLD’’), and at least
one of the most commonly used drugs: ‘‘penicillamine,’’
‘‘trientine,’’ ‘‘zinc,’’ ‘‘tetrathiomolybdate’’, and their brand
names, acronyms, abbreviations, and synonyms. Inclusion
criteria included prospective, retrospective, randomized,
and non-randomized controlled studies with human sub-
jects published as full articles written in English or Chi-
nese. Exclusion criteria included animal studies, case
reports or case series, reviews, letters, short papers, edito-
rials, metal metabolism or pharmacological research,
diagnostic or other testing studies, liver transplants or other
non-medication treatments, duplicate reports, and insuffi-
cient data. Date of last search: September 1, 2014. 2.3 Treatment Effect Definition To maintain consistency, we used a comparative unit called
‘‘treatment block’’ (TB) [16] to calculate the frequencies of
adverse effects and treatment effectiveness for patients dur-
ing specific time durations. One TB equaled the duration of
one therapy up to the time that a medication was changed, or
until the end of the follow-up period [16]. For our purposes, a
TB was considered effective when the author of a research
paper used terms such as ‘‘effective,’’ ‘‘efficacious,’’ ‘‘suc-
cessful,’’ ‘‘improved,’’ ‘‘stable,’’ ‘‘normal,’’ ‘‘biochemically
improved,’’ ‘‘responsive,’’ ‘‘non-progressive’’, or their syn-
onyms. The list of terms indicating non-effectiveness inclu-
ded ‘‘ineffective,’’ ‘‘inefficacious,’’ ‘‘failed,’’ ‘‘deteriorated,’’
‘‘worsened,’’ ‘‘degenerated,’’ ‘‘abnormal,’’ ‘‘severe side
effects,’’ ‘‘LT,’’ ‘‘dead,’’ ‘‘treatment failure,’’ ‘‘stationary,’’
‘‘unchanged,’’ ‘‘clinical suspicion,’’ ‘‘progressive,’’ ‘‘non-
responsive’’, or their synonyms. 2.2 Definition of WD Phenotypes optimal medication therapy. One reason is the diversity of
WD genotypes and phenotypes, which makes it difficult to
determine differences in
drug
effectiveness. Another
challenge is the small number of known cases, with a
worldwide prevalence of 1/30,000 [18]. This makes it
difficult to conduct large-scale cohort randomized clinical
trials. Our motivations were to collect and compile avail-
able data from past studies on the effectiveness and safety
of commonly used WD medications, and to review original
studies found in the PubMed database. Four phenotype presentation categories were noted: neu-
rological, hepatic, mixed, and asymptomatic. Following the
lead of Ferenci et al. [36], patients with neurological and/or
psychiatric symptoms at diagnosis were classified as neu-
rological. The definition of hepatic presentation required
the exclusion of neurological symptoms noted during a
detailed examination at the time of diagnosis [36]. Pure
hematological abnormalities such as Coombs negative
hemolytic anemia were classified as hepatic. Asymp-
tomatic manifestations included asymptomatic and pre-
symptomatic presentation. Finally, patients with other
miscellaneous symptoms (e.g., renal dysfunction and bone
deformities) were placed in a mixed presentation category
with patients showing simultaneous hepatic and neu-
ropsychiatric symptoms. In combination therapies for WD, zinc and a chelating
agent (chelator) are utilized to block copper uptake and to
eliminate excess copper [16]. The two medications must be
taken at least 1 hour apart in order to mitigate zinc
chelation [14]. They are still considered controversial, with
few rigorously designed studies and little in the way of
safety data [29]. The most frequently cited studies that
suggest favorable outcomes for combination therapy using
DPA plus zinc or TETA plus zinc [29] are those by Dha-
wan et al. [30], Askari et al. [31], and Santos Silva et al. [32]. However, some earlier studies [33, 34] reported no
advantages for the DPA-zinc combination. Both EASL
[19] and AASLD [20] assert that considerably more
research is required to determine whether combination
therapy using a chelator plus zinc has advantages for WD
patients. Therefore, our primary goal was to verify whether
combination therapies are effective and safe at statistically
significant
levels
for
patients
with
different
clinical
presentations. 1 Introduction Wilson’s disease (WD) (also known as hepatolenticular
degeneration, or HLD) is a rare inherited autosomal
recessive disorder associated with mutations in the ade-
nosine triphosphatase 7B (ATP7B) gene [1–18] and char-
acterized by copper metabolic abnormalities [19, 20]. Excessive copper accumulation can result in toxicity and
damage to the brain, liver, kidney, and other tissues. WD
has a broad spectrum of clinical presentations, with hepatic
and neurological symptoms considered the main features
[21]. While liver transplants (LTs) and gene therapies are
provided to a small number of WD patients, the large
majority
require
lifelong
medication
to
control
the
absorption and storage of copper in their bodies. The most
commonly used drugs are penicillamine (DPA) [22], tri-
entine
(TETA)
[23], zinc
salts
(Zn)
[24,
25], and
tetrathiomolybdate (TM; an experimental therapy that is
not yet commercially available) [26, 27]. The goal of
medication is to prevent, stabilize, or reverse copper
overload and WD symptoms [28]. The European Association for the Study of the Liver
(EASL) [19] and the American Association for the Study
of Liver Diseases (AASLD) [20] announced their respec-
tive clinical practice guidelines for WD in 2012 and 2008,
but no clear international consensus exists regarding an 4
Department of Internal Medicine, Ben Tang Cheng Ching
Hospital, Taichung 41364, Taiwan 12 3 3 698 J.-C. Chen et al. J.-C. Chen et al. 2.5 Statistical Analyses retrieved titles and abstracts, and 245 were excluded during
the tertiary step of scanning the full texts of potentially
eligible studies. A total of 50 studies were included for
prevalence investigation, and of these, 17 described out-
comes from combination therapies and were therefore
accepted for this review [1–17]. The study selection pro-
cedure is summarized in Fig. 1, characteristics of the 17
studies are shown in Table 1, and mean follow-up times
and outcomes regarding the effectiveness of combination
therapies in each study are shown in Table 2. As shown,
the papers in the final sample discussed seven combina-
tions of a chelator and a zinc salt: (a) DPA ? Zn sulfate,
(b) DPA ? unknown or another Zn salt (e.g., zinc glu-
conate), (c) TETA ? Zn sulfate, (d) TETA ? Zn acetate,
(e) TETA ? unknown or another Zn salt, and (f) unknown
chelator DPA or TETA ? any Zn salt. All analyses were performed using Cochrane RevMan 5.3
and SPSS Statistics 22.0. Pooled relative risk ratios (RRs)
and 95 % confidence intervals (CIs) were calculated from
the original study data using the Mantel–Haenszel method
with a random-effects model. A fixed-effects model was
selected for cases with low heterogeneity (I2 \ 30 %). The
Mantel–Haenszel method generates estimates of associa-
tions between exposures and outcomes after accounting for
confounding effects. We stratified the data into two or
more confounding factor levels before computing pooled
RRs across the strata. Note that the random-effects model
has a stricter assumption than the fixed-effects model. We
used the random-effects model to achieve conservative RR
and CI estimates. The Mantel–Haenszel equation for an RR is: RR ¼
X
m
j¼1
ajðcj þ djÞ
nj
,X
m
j¼1
cjðaj þ bjÞ
nj
ð1Þ ð1Þ bers of patients in each
j-th stratum of the con-
e number of patients in
number of strata. tween discrete values of
erent treatment groups
square (v2) test and Phi
ed Chi square tests to
endence between effec-
dications/phenotypes. A
some degree of corre-
To obtain measures of
es, Phi or Cramer’s V
e of between 0 and 1. A
a maximum numerical
ad a perfect relationship
0 when there was no
measure of association
measure was significantly
s showing a significant
bles. A p value of\0.05
cant. 2.5 Statistical Analyses ed, 139 of which were
in languages other than
studies were excluded
process of reading the
916 potenally relevant studies
idenfied and screened for retrieval
139 studies were excluded as in
language other than English and
Chinese
194 case reports were
excluded
87 animal studies were
excluded
777 studies screened on
tle/abstract
199 studies were excluded as
reviews, leers, short papers, or
editorials
297 studies screened on full text
153 studies were excluded:
65 on metal metabolism or
pharmacological research
54 on diagnoses, genec assays,
assessments, or other tests
34 on liver transplantaons or
other non-medicaon treatments
94 studies were excluded:
55 with data duplicaon, overlap,
or deficiency
39 on dermal lesions, pregnancy,
other diseases, or the
experimental tetrathiomolybdate
50 studies included for
prevalence invesgaon
17 studies on combinaon therapy
included for this review
Fig. 1 Flow chart of included and excluded studies for this review where aj, bj, cj, and dj are the numbers of patients in each
cell of a two-by-two table in the j-th stratum of the con-
founding variable, nj represents the number of patients in
the j-th stratum, and m is the total number of strata. 139 studies were excluded as in
language other than English and
Chinese Correlations and associations between discrete values of
nominal data variables from different treatment groups
were evaluated using a Pearson Chi square (v2) test and Phi
or Cramer’s V measures. We used Chi square tests to
determine the likelihood of independence between effec-
tiveness/safety and different medications/phenotypes. A
rejected null hypothesis suggested some degree of corre-
lation between the two variables. To obtain measures of
association between those variables, Phi or Cramer’s V
values were calculated using a value of between 0 and 1. A
measure of association achieved a maximum numerical
value of 1 when the two variables had a perfect relationship
with each other, and a value of 0 when there was no
relationship. After the observed measure of association
values had been calculated, if the measure was significantly
different from 0, it was viewed as showing a significant
relationship between the two variables. A p value of\0.05
was considered statistically significant. 2.4 Data Extraction To select studies according to our inclusion and exclusion
criteria, two authors initially screened the entry informa-
tion, titles, and abstracts of all retrieved records. Next, full
texts were scanned to determine conformance with the
criteria. Two authors independently extracted data and
outcomes using a standardized form. All disagreements
were discussed with a third author. Studies were included
and data extracted in cases where a consensus was
achieved. Other extracted information included first author,
country, publication year, number of patients, patient
gender ratio, patient phenotype ratio, adverse effects, and
mortality (liver transplant and deceased counts). 12 123 123 Combination Therapy Using Chelating Agent and Zinc for Wilson’s Disease 699 3.2 Prevalence Investigation 75.4–89.7 %)) [37], and Weiss et al. for zinc monotherapy
(63/88, or 71.6 % (95 % CI 62.2–81.0 %)) [16]. As shown
in Table 3, results from our inter-study analysis indicate
significant differences in effectiveness rates between
combination therapies and the three monotherapies: an RR
of 0.82 for DPA [15] (95 % CI 0.71–0.94, Fig. 2), an RR of
0.73 for TETA [37] (95 % CI 0.65–0.82, Fig. 3), and an
RR of 0.84 for Zn [16] (95 % CI 0.72–0.98, Fig. 4). In this
part of our study, we used the number of effective TBs as
the number of events. Of the 2954 WD patients mentioned in 45 of the 50 studies
included for the prevalence investigation, 1357 (45.9 %)
were female (95 % CI 44.1–47.7 %). Pooled mean age at
diagnosis as mentioned in 47 of the same 50 papers was
18.7 years, ranging from 6 to 40 years. For our phenotype
prevalence investigation, of the 2988 patients mentioned in
47 of the 50 included studies, those with neurological,
hepatic, mixed, or asymptomatic presentations numbered
1058 (35.4 %), 1242 (41.6 %), 341 (11.4 %), and 347
(11.6 %), respectively. The number of hepatic patients was
approximately 1.2 times that of neurological patients. When combined with the mixed phenotype, the total
number of hepatic patients (i.e., at least one liver-related
symptom) was 1583 (52.9 % of the total patient sample). We then searched for relationships between phenotype
and combination therapy effectiveness, and found that less
than one half (47.1 %, 95 % CI 38.2–56.0 %) of the TBs in
the hepatic group (mixed phenotype excluded) responded
well to combination therapy, compared to 78.6 % (95 % CI
70.7–86.6 %) of TBs in the neurological group (Table 2). According
to
our
subgroup
analyses
(two-phenotype
stratification), a statistically significant difference exists
between the two subgroups (p = 0.02) (Fig. 5). The RR of
the
overall
effectiveness
rate
was
0.63
(95 %
CI
0.43–0.94), indicating that the combination therapies were
31.5 % (95 % CI 18.8–44.3 %) less effective for the hep-
atic patients than for the neurological patients. Note that
the total number of TBs involving patients in different
phenotype groups does not equal the overall effectiveness 3.1 Included Literature asymptomatic or presymptomatic, – not available Table 1 Characteristics of 17 included studies on combination therapies 3.1 Included Literature A total of 916 hits were screened, 139 of which were
excluded because they were written in languages other than
English or Chinese. Another 480 studies were excluded
during the secondary selection process of reading the 50 studies included for
prevalence invesgaon Fig. 1 Flow chart of included and excluded studies for this review 12 3 J.-C. Chen et al. 700 Table 1 Characteristics of 17 included studies on combination therapies
PMID
Study
Country
Mean
age
(years)
No. of
patients
Gender
Phenotype
Data collected
Male
Female
Nue. Hep. Mixed
Asy. From
To
23011036
Sini et al. [1]
Italy
23.0
60
19
41
0
38
22
0
1981
2011
22055589
El-Karaksy et al. [2]
Egypt
10.3
54
31
23
5
33
3
13
1996
2009
22355993
Noureen and Rana [3]
Pakistan
9.1
50
34
16
46
0
4
0
2005
2008
21682854
Abdel et al. [4]
Egypt
10.0
77
43
34
6
35
9
27
1992
2009
17709362
Sinha et al. [5]
India
14.4
50
30
20
39
3
8
0
1999
2002
14759316
Li et al. [6]
China
10.0
21
9
12
6
9
6
0
1990
1998
11837754
Sinha et al. [7]
India
13.3
49
38
11
27
0
22
0
1991
2000
10869138
Kalra et al. [8]
India
7.2
25
14
11
5
7
9
4
1986
1997
10745386
Yu¨ce et al. [9]
Turkey
9.0
34
19
15
4
30
0
0
1980
1998
9193846
Schumacher et al. [10]
Germany
27.0
13
5
8
4
7
2
0
1988
1995
8076990
Gill et al. [11]
India
19.6
11
7
4
0
11
0
0
–
–
11819363
Ren et al. [12]
China
19.0
120
65
55
–
–
–
–
1994
1997
16606763
Brewer et al. [13]
USA
–
23
–
–
23
0
0
0
–
–
17460493
Arnon et al. [14]
USA
12.5
22
11
11
0
15
2
0
1998
2006
20958917
Bruha et al. [15]
Czech
38.5
117
59
58
21
51
34
11
1965
2008
21185835
Weiss et al. [16]
Germany and
Austria
17.5
288
123
165
60
157
39
32
1954
2008
24661374
Ranucci et al. [17]
Italy
6.0
42
30
12
0
38
4
0
1984
2012
Total
17.6
1056
537
496
246
434
164
87
Percentage (%)
(mean)
52.0
48.0
26.4
46.6
17.6
9.3
PMID PubMed literature ID, Neu. neurological, Hep. hepatic, Asy. 3.3 Effectiveness 702 Table 3 Effectiveness rates and RRs between combination therapies and three monotherapies as found in literature
No. of
TBs
Effective
TBs
Effective
rate (%)
Effective
95 % CI (%)
RR between combination
therapies and others
On combination therapies (pooled)
437
264
60.4
55.8–65.0
–
On DPA monotherapy [15]
99
73
73.7
65.1–82.4
0.82
On TETA monotherapy [37]
109
90
82.6
75.4–89.7
0.73
On zinc monotherapy [16]
88
63
71.6
62.2–81.0
0.84
TBs treatment blocks, CI confidence interval, RR relative risk ratio, DPA penicillamine, TETA trientine
Fig. 2 Forest plot of 17 included studies measuring relative risk of pooled effectiveness following combination therapy compared to DPA
monotherapy [15] Table 3 Effectiveness rates and RRs between combination therapies and three monotherapies as found in literature
No. of
TBs
Effective
TBs
Effective
rate (%)
Effective
95 % CI (%)
RR between combination
therapies and others
On combination therapies (pooled)
437
264
60.4
55.8–65.0
–
On DPA monotherapy [15]
99
73
73.7
65.1–82.4
0.82
On TETA monotherapy [37]
109
90
82.6
75.4–89.7
0.73
On zinc monotherapy [16]
88
63
71.6
62.2–81.0
0.84
TBs treatment blocks, CI confidence interval, RR relative risk ratio, DPA penicillamine, TETA trientine e 3 Effectiveness rates and RRs between combination therapies and three monotherapies as found in literature Fig. 2 Forest plot of 17 included studies measuring relative risk of pooled effectiveness following combination therapy compared to DPA
monotherapy [15] Fig. 2 Forest plot of 17 included studies measuring relative risk of pooled effectiveness following combination therapy compared to DPA
monotherapy [15] Fig. 2 Forest plot of 17 included studies measuring relative risk of pooled effectiveness following combination therapy compared to DPA
monotherapy [15] Fig. 3 Forest plot of 17 included studies measuring relative risk of pooled effectiveness following combination therapy compared to TETA
monotherapy [37] Fig. 3 Forest plot of 17 included studies measuring relative risk of pooled effectiveness following combination therapy compared to TETA
monotherapy [37] Fig. 4 Forest plot of 17 included studies measuring relative risk of pooled effectiveness following combination therapy compared to Zn
monotherapy [16] Fig. 4 Forest plot of 17 included studies measuring relative risk of pooled effectiveness following combination therapy compared to Zn
monotherapy [16] 3.3 Effectiveness Of the 437 pooled TBs shown in Table 2, 264 responded
positively to a combination therapy, for an overall effec-
tiveness rate of 60.4 % (95 % CI 55.8–65.0 %), lower than
the rates reported by Bruha et al. for DPA monotherapy
(73/99, or 73.7 % (95 % CI 65.1–82.4 %)) [15], Weiss
et al. for TETA monotherapy (90/109, or 82.6 % (95 % CI 123 Combination Therapy Using Chelating Agent and Zinc for Wilson’s Disease 701 Table 2 Outcomes of 17 included studies on combination therapy effectiveness for WD
PMID
Mean
follow-up
Comb. type
Total dose (mg/d) used
Comb. TB#
Stratified by phenotype
Overall effectiveness
Neurological
Hepatic
Mixed
Asymptomatic
Chelator
Zn
NE? NE-
HE? HE-
ME? ME-
AE? AE-
E? E-
23011036
25.0
a
600–1200
150–300
13
6
2
1
4
7
6
22055589
2.0
a
40/kg
75–150
41
4
1
10
23
2
1
16
25
22355993
3.0
a
10–30/kg
50–100
50
46
4
46
4
21682854
4.9
a
18/kg(mean)
50–150
58
2
2
11
16
3
5
11
8
27
31
17709362
4.1
a
–
–
50
36
14
14759316
–
a
10–30/kg
67.5
18
9
2
13
5
13
5
11837754
3.8
a
500–1000
100
18
5
13
10869138
–
a
8–30/kg
50
20
13
7
10745386
9.0
a
20/kg
68.2
26
4
0
11
11
15
11
9193846
–
a
–
–
6
0
4
0
2
0
6
8076990
–
a
500–1000
100–150
6
4
2
4
2
–
c
500
100
3
2
1
2
1
11819363
2.8
b
1000
80
60
35
25
16606763
1.0
e
1000
100–150
23
16
7
16
7
17460493
2.5
d
500–1000
50–100
2
0
2
0
2
20958917
15.1
b
600–1200
150
2
0
2
0
2
21185835
17.1
f
–
–
30
22
8
24661374
12.0
b
750–1500
50–150
11
7
4
Mean/Total
10.6
437
81
22
57
64
6
10
11
8
264
173
Percentage (%)
years
78.6
21.4
47.1
52.9
37.5
62.5
57.9
42.1
60.4
39.6
Comb. type (combination therapy type): a DPA ? Zn sulfate, b DPA ? Zn, c TETA ? Zn sulfate, d TETA ? Zn acetate, e TETA ? Zn, f chelator ? Zn; Comb. TB# number of treatment
blocks on combination therapy; Zn zinc E? effective E- ineffective; – not available
rapy Using Chelating Agent and Zinc for Wilson s Disease 12 J.-C. Chen et al. 3.4 Adverse Effects number since some of the studies in the sample did not give
specific statistics for different phenotypes. A comparison of number since some of the studies in the sample did not give
specific statistics for different phenotypes. A comparison of
all phenotypes and combination therapy effectiveness
revealed statistically significant correlations between the
two factors (v2(3) = 26.666, p \ 0.001) (data not shown);
medium–low positive correlations between the two vari-
ables were noted in the form of Cramer’s V value (0.321,
significant at 0.001). In contrast, results from correlation
and difference tests involving various combination therapy
types and overall effectiveness were not statistically sig-
nificant
(v2(1) = 0.373,
p = 0.541
and
Z = -0.611,
p = 0.271). In other words, the data indicate that similar
results are produced by all of the combination therapy
types reviewed for this paper. number since some of the studies in the sample did not give
specific statistics for different phenotypes. A comparison of
all phenotypes and combination therapy effectiveness
revealed statistically significant correlations between the
two factors (v2(3) = 26.666, p \ 0.001) (data not shown);
medium–low positive correlations between the two vari-
ables were noted in the form of Cramer’s V value (0.321,
significant at 0.001). In contrast, results from correlation
and difference tests involving various combination therapy
types and overall effectiveness were not statistically sig-
nificant
(v2(1) = 0.373,
p = 0.541
and
Z = -0.611,
p = 0.271). In other words, the data indicate that similar
results are produced by all of the combination therapy
types reviewed for this paper. Data on combination therapy safety, including adverse
effects and mortality, are presented in Table 4. Since the
first combination therapy type was clearly the most com-
mon, we collapsed the other six to create a workable bal-
ance between sample sizes. Note that we split the statistics
for one study [11] into two parts because the patients were
treated with two different combination therapies. Since
some of the studies in the sample did not specifically
describe adverse reactions for different phenotypes, the
numbers of TBs for different phenotypes and for overall
adverse effects are not equal. Of the 271 TBs listed in
Table 4, 97 resulted in adverse reactions, an overall 123 123 703 Combination Therapy Using Chelating Agent and Zinc for Wilson’s Disease yp
py yp ,
Mortality (Dead ? LT)/(Dead ? LT ? Alive) Combination Therapy Using Chelating Agent and Zinc for Wilson’s Disease Alive) Table 4 Safety investigation data for combination therapies in analyzed studies 12 3 704 J.-C. Chen et al. four most commonly used medications (DPA, TETA and
Zn monotherapies, and combination therapy) and from
studies of the combination therapies alone are presented in
Table 5. As shown, of the 2239 patients mentioned in 44 of
the 50 studies of the four most common medications, 103
died and 44 received liver transplants, a mortality rate of
6.6 % (95 % CI 5.5–7.6 %). In contrast, the mortality rate
for all patients receiving some form of combination therapy
was 12.7 % (95 % CI 9.5–15.9 %), suggesting that those
therapies failed to help a large number of individuals with
acute WD. For this part of our analysis, we used the
number of deceased and liver transplant patients as the
number of events. A statistically significant difference was
found between mortality rates for patients receiving com-
bination therapies and those receiving common medica-
tions (Fig. 7; RR: 1.94, p \ 0.001). Since we did not
measure the percentages of patients who experienced acute
liver failure, the two group analyses may suffer from bias. Still, the pooled data suggest that combination therapy
patients had a much higher mortality rate compared to
those receiving the other frequently used medications. adverse effect rate of 35.8 % (95 % CI 30.1–41.5 %). The
percentage for patients in the hepatic category was 41.7 %
(95 % CI 31.1–52.2 %) and that for those in the neuro-
logical category was 26.3 % (95 % CI 12.3–40.3 %), not
significantly different (p = 0.84), perhaps due to the small
sample size. Results from our analysis of inter-studies on
adverse effect rates are presented in Fig. 6, with the
number of events noted as the number of TBs presenting
adverse effects. The data indicate that the combination
therapies resulted in greater relative risk compared to those
for the TETA (RR: 1.67, 95 % CI 1.04–2.69) and Zn (RR:
2.25, 95 % CI 1.36–3.73) monotherapies [16], but not that
for the DPA monotherapy [37] (RR: 1.10, 95 % CI
0.87–1.38). Statistically significant differences were not
noted for correlation and difference measures between
different combination therapy types and overall adverse
effects
(v2(1) = 0.938,
p = 0.333
and
Z = -0.968,
p = 0.166). Combination Therapy Using Chelating Agent and Zinc for Wilson’s Disease Fig. 5 Forest plot of studies on combination therapies for hepatic phenotype versus neurological phenotype examining relative risk of
effectiveness Fig. 5 Forest plot of studies on combination therapies for hepatic phenotype versus neurological phenoty
effectiveness Fig. 5 Forest plot of studies on combination therapies for hepatic phenotype versus neurological phenotype examining relative risk of
effectiveness Fig. 5 Forest plot of studies on combination therapies for hepatic phenotype versus neurological phenotype examining relative risk of
effectiveness Table 4 Safety investigation data for combination therapies in analyzed studies
PMID
Comb. type
Phenotype
Overall adverse
effects
Mortality on combination
therapy
Neurological
Hepatic
Mixed
Asymptomatic
NA? NA-
HA? HA-
MA? MA-
AA? AA-
A? A-
Dead
LT
Alive
Mortality
(%)
23011036
a
0
0
13
0.0
22055589
a
1
4
10
23
1
2
12
29
8
3
30
26.8
22355993
a
0
0
50
0.0
21682854
a
4
0
18
9
7
1
3
16
32
26
16
0
42
27.6
17709362
a
0
0
50
0.0
14759316
a
4
14
5
0
13
27.8
11837754
a
–
–
–
–
10869138
a
1
0
19
5.0
10745386
a
0
4
7
15
7
19
4
3
19
26.9
9193846
a
0
6
0
100.0
8076990
a
1
0
5
16.7
Subtotal
5
8
35
47
8
3
3
16
55
88
35
12
241
16.3
Percentage (%)
38.5
61.5
42.7
57.3
72.7
27.3
15.8
84.2
38.5
61.5
12.2
4.2
83.7
8076990
c
1
0
2
33.3
11819363
b
22
38
0
0
60
0.0
16606763
e
5
18
5
18
4
0
19
17.4
17460493
d
0
2
0
2
0
0
2
0.0
20958917
b
0
2
0
2
–
–
–
–
21185835
f
10
20
0
1
29
3.3
24661374
b
5
6
0
0
11
0.0
Subtotal
5
20
0
2
0
0
0
0
42
86
5
1
123
4.7
Percentage (%)
20.0
80.0
0.0
100
–
–
–
–
32.8
67.2
3.9
0.8
95.3
Total
10
28
35
49
8
3
3
16
97
174
40
13
364
12.7
Percentage (%)
26.3
73.7
41.7
58.3
72.7
27.3
15.8
84.2
35.8
64.2
9.6
3.1
87.3
Comb. type combination therapy type, A? adverse effect, A- non-adverse effect, – not available, LT liver transplantation, Alive LT excluded,
Mortality (Dead ? LT)/(Dead ? LT ? 3.5 Mortality Detailed mortality data associated with combination ther-
apy studies are presented along the right-hand side of
Table 4. Pooled results from mortality investigations of the Mortality rates for patients in different phenotype
groups were difficult to determine due to the small sample
size and lack of mortality data for each group. However, Fig. 6 Forest plot of 17 included studies measuring relative risk of pooled adverse effects following combination therapy compared to a DPA,
b TETA, and c Zn monotherapies Fig. 6 Forest plot of 17 included studies measuring relative risk of pooled adverse effects following combination therapy compared to a DPA,
b TETA, and c Zn monotherapies cluded studies measuring relative risk of pooled adverse effects following combination therapy compared to a DPA
erapies Table 5 Mortality statistics for different patient groups in included studies
No. of patients (%)
Dead
LT
Alive
Mortality (%)
Patients in total
Studies included
On common medications (pooled)
103 (4.6 %)
44 (2.0 %)
2092 (93.4 %)
6.6
2239
44 of 50
On combination therapies (pooled)
40 (9.6 %)
13 (3.1 %)
364 (87.3 %)
12.7
417
15 of 17
LT liver transplantation, Mortality (Dead ? LT)/(Patients in total) 12 Combination Therapy Using Chelating Agent and Zinc for Wilson’s Disease 705 Fig. 7 Forest plot of 17 included studies measuring relative risk of pooled mortality following combination therapy compared to pooled
mortality of four most commonly used medications Fig. 7 Forest plot of 17 included studies measuring relative risk of pooled mortality following combination therapy compared to pooled
mortality of four most commonly used medications we did find statistically significant differences in mortality
among patients receiving different types of combination
therapies (p \ 0.001). Patients in the DPA ? Zn sulfate
group had a much higher mortality rate compared to those
in all other groups (16.3 vs. 4.7 %; RR: 3.51, 95 % CI
1.54–8.00). Chi square test results indicate a statistically
significant
correlation
(v2(1) = 10.933;
p = 0.001)
between combination therapy type and mortality. we did find statistically significant differences in mortality
among patients receiving different types of combination
therapies (p \ 0.001). Patients in the DPA ? Zn sulfate
group had a much higher mortality rate compared to those
in all other groups (16.3 vs. 4.7 %; RR: 3.51, 95 % CI
1.54–8.00). 3.5 Mortality Chi square test results indicate a statistically
significant
correlation
(v2(1) = 10.933;
p = 0.001)
between combination therapy type and mortality. patients receiving DPA plus zinc sulfate compared to other
types of combination therapies (16.3 vs. 4.7 %). Thus,
Yonetani and Walshe [34] emphasize the danger of using
zinc sulfate with any chelation regimen. y
g
From our analysis, it appears that the literature lacks
rigorously designed studies and safety data on combination
therapies using zinc and a chelator. The three most fre-
quently cited studies that suggest favorable outcomes for
combination therapies involving zinc plus either DPA or
TETA are those by Dhawan et al. [30], Askari et al. [31],
and Santos Silva et al. [32]. Dhawan et al.’s research focus
was the scoring system for WD liver transplants [30], but in
their report they claim that 20 symptomatic, non-deceased,
and non-liver-transplant-receiving children did not require
transplants for a long period of time after receiving a
combination of DPA plus zinc. Their study is lacking in
several respects: it does not include detailed evaluations
regarding the clinical effectiveness of combination therapy,
nor do they provide follow-up information for seven
asymptomatic siblings who were treated with a combina-
tion of DPA and zinc. In their paper, Askari et al. [31]
described the successful use of TETA plus zinc in eight
patients
with
decompensated
hepatic
WD,
but
their
approach involved the use of that combination therapy for
4 months, followed by a regimen of zinc monotherapy. They claim that the combination therapy reduced or
eliminated the need for liver transplants, but the time
period involved was imprecise. Santos Silva et al. [32]
evaluated the effectiveness of DPA plus zinc for treatment
periods ranging from one to 2 years, but some of the
patients in their study had to be shifted to other therapies
due to the adverse effects of the initial combination ther-
apy. They mention three combination therapy patients
during an initial follow-up period and four during a second
follow-up period, but they are unclear about overlaps. 4 Discussion In light of the rarity of WD cases and lack of clinical
consensus on the best medications, our goal was to sys-
tematically review the literature and to perform statistical
analyses to support or refute assertions of the success of
various combination therapies. Our results indicate a suc-
cess rate for combination therapies of approximately 60 %,
much lower than expected. That figure is significantly less
than those reported by Bruha et al. for DPA monotherapy
(73.7 %) [15], Weiss et al. for TETA monotherapy
(82.6 %) [37], and Weiss et al. for zinc monotherapy
(71.6 %) [16]. We found strong evidence indicating that
hepatic patients do not respond well to combination ther-
apies, with a reported effectiveness rate of 47.1 versus
78.6 % for patients with neurological manifestations. Results from a pooled analysis show that compared to
hepatic manifestation patients, neurological patients were
significantly more likely to receive benefits from combi-
nation therapies. For example, Pellecchia et al. [38] found
that DPA combined with zinc is effective and safe for
neurologically impaired patients. In terms of safety, the
studies we reviewed reported a 35.8 % pooled adverse
effect for all patients receiving some type of combination
therapy (41.7 % for hepatic patients and 26.3 % for
patients with neurological presentations). The lack of a
statistically significant difference between the two pheno-
types is likely due to the small sample size, yet there is still
potential for clinical significance. Regarding mortality
associated with combination therapies, the 12.7 % rate was
significantly higher than the 6.6 % rate reported for com-
mon medication therapies. It is likely that the mortality rate
is higher for hepatic patients, but the reviewed studies did
not contain specific mortality data for that group. One
unexpected finding was the higher mortality rate for Some researchers [33, 34] have argued that there is no
advantage to the concomitant administration of DPA and
zinc, suggesting that zinc may interact with both DPA and
TETA, and possibly inhibit chelator absorption and action
[33, 39]. A third research team has made the strong rec-
ommendation that zinc sulfate should never be used with
chelation medication [34]. Friedman and Yarze [40] also
argue that it is counterproductive to use a combination of
chelators and zinc in WD patients. According to EASL
guidelines [19], there are no known advantages to using 12 3 3 J.-C. Chen et al. 4 Discussion 706 patients receiving a combination therapy was 12.7 %,
double that reported for patients receiving the four most
commonly used medications. Another important finding is
that the combination therapy of DPA plus zinc sulfate
resulted in much higher mortality rates compared to those
for all other combination therapy types (16.3 vs. 4.7 %). However, the pooled data cannot be considered high-
quality evidence for estimating the effectiveness and safety
of combination therapies. Thus, these findings should be
used to support treatment decisions only until more and
higher-quality evidence becomes available. More large-
cohort randomized clinical trials and/or evidence-based
studies are still required to fully address the issues men-
tioned in this review. Our primary conclusion is that clin-
icians should closely monitor biochemical test results and
clinical courses for WD patients receiving combination
therapies, especially in response to hepatic manifestations. combination therapies involving a chelator and zinc,
though they do not deny the possibility. AASLD guidelines
[20] are unclear on this question, simply stating a need for
more confirmatory research. In one retrospective cohort study [16], six combination
therapy TBs were discontinued because the physician
suspected that the zinc and chelator were interacting
pharmacologically. Arnon et al. [14] reported that two
patients taking TETA monotherapy alone for 6 months had
their hepatic alanine aminotransferase (ALT) levels return
to normal. The decision was made to switch to a combi-
nation of TETA plus zinc, but after another 6 months their
ALT levels nearly doubled, and after a full year they were
almost three times the level considered normal [14]. The
authors speculated that the patients may not have been
adherent, but this idea was neither tested nor verified. They
did, however, suggest that future combination therapy was
unnecessary. The
literature
contains
other
evidence
concerning
chelator-zinc interaction. In their study of urinary copper
excretion following TETA monotherapy, Dubois et al. [41]
reported that urinary zinc content increased from 181
lg/day pre-treatment to 402 lg/day post-treatment. Their
observations were similar to those reported by McCall
et al. [42] in a metabolic study involving DPA trials. Kodama et al. [43] reported a significant increase in the
urinary excretion of zinc in a group of healthy (non-WD)
volunteers during the first 6 hours following TETA
administration. 6 Limitations Possible limitations to our findings include a lack of
stratification for mild, moderate, and severe adverse
effects, plus the apparent lack of high-quality evidence in
support of estimates of relative effectiveness and adverse
effects of combination therapies versus monotherapies. Further, there may be bias in some interpretations of results
due to the lack of substantial data and additional reports on
combination therapies. For these reasons, no firm recom-
mendations can be drawn from the pooled data. Note also
that WD is an intractable disease, meaning that individual
patients may have different responses to each of the four
most commonly used medications due to variance in WD
genotypes and phenotypes. Consequently, neither a stan-
dard treatment regimen nor a clear consensus exists
regarding an optimal medication therapy for treating the
disease. More evidence-based studies and/or large-cohort
randomized controlled comparative trials are required. Acknowledgments
The authors gratefully acknowledge the finan-
cial support from the Taiwan Ministry of Science and Technology
(NSC 102-2221-E-468-010- and MOST 103-2221-E-468-002-). Open Access
This article is distributed under the terms of the
Creative Commons Attribution 4.0 International License (http://crea
tivecommons.org/licenses/by/4.0/), which permits unrestricted use,
distribution, and reproduction in any medium, provided you give
appropriate credit to the original author(s) and the source, provide a
link to the Creative Commons license, and indicate if changes were
made. 5 Conclusion Our main findings are (a) an overall effectiveness rate of
only 47.1 % and (b) an overall adverse effect rate of
41.7 % among hepatic patients treated with combination
therapies. We also found that the overall mortality rate for 4 Discussion Kuchinskas and Rosen [39] investigated the
affinities of bivalent metals for DPA, and reported a high-
to-low affinity order of Hg [ Ni [ Cu [ Zn [ Cd [ Pb;
this serves as indirect evidence that DPA is capable of
chelating both copper and zinc. Cossack and Bouquet [44]
have described a sub-clinical deficiency of zinc induced by
DPA treatment. In an animal study [45], Fieten et al. evaluated hepatic copper and zinc concentrations before
and after DPA monotherapy treatment in 42 Labrador
Retrievers, and reported significant decreases in both
concentrations in the dogs’ livers. Combined, these studies
suggest that zinc should not be combined with a chelator,
even several hours apart, because doing so is likely to
reduce the effectiveness of zinc for treating WD. Further,
the existing evidence indicates that the presence of zinc in
the bloodstream and gut may alter the effect of chelators on
copper. 1. Sini, M., Sorbello, O., Sanna, F., Battolu, F., Civolani, A., Fanni,
D., et al. (2013). Histologic evolution and long-term outcome of References Treatment of Wilson’s
disease with penicillamine and zinc salts: A follow-up study. Chinese Journal of Pediatrics, 41, 119–122. 7. Sinha, S., Jha, D. K., & Sinha, K. K. (2001). Wilson’s disease in
Eastern India. Journal of the Association of Physicians of India,
49, 881–884. 26. Walshe, J. M. (1984). Copper: Its role in the pathogenesis of liver
disease. Seminars in Liver Disease, 4, 252–263. 8. Kalra, V., Khurana, D., & Mittal, R. (2000). Wilson’s disease-
early onset and lessons from a pediatric cohort in India. Indian
Pediatrics, 37, 595–601. 27. Brewer, G. J., Hedera, P., Kluin, K. J., Carlson, M., Askari, F.,
Dick, R. B., et al. (2003). Treatment of Wilson disease with
ammonium tetrathiomolybdate: III. Initial therapy in a total of 55
neurologically affected patients and follow-up with zinc therapy. Archives of Neurology, 60, 379–385. 9. Yu¨ce, A., Kocak, N., Gu¨rakan, F., & Ozen, H. (2000). Wilson’s
disease with hepatic presentation in childhood. Indian Pediatrics,
37, 31–36. 28. Wiggelinkhuizen, M., Tilanus, M. E., Bollen, C. W., & Houwen,
R. H. (2009). Systematic review: Clinical efficacy of chelator
agents and zinc in the initial treatment of Wilson disease. Ali-
mentary Pharmacology & Therapeutics, 29, 947–958. 10. Schumacher, G., Platz, K. P., Mueller, A. R., Neuhaus, R., Stein-
mu¨ller, T., Bechstein, W. O., et al. (1997). Liver transplantation:
Treatment of choice for hepatic and neurological manifestation of
Wilson’s disease. Clinical Transplantation, 11, 217–224. 29. Weiss, K. H., & Stremmel, W. (2012). Evolving perspectives in
Wilson disease: Diagnosis, treatment and monitoring. Current
Gastroenterology Reports, 14, 1–7. 11. Gill, H. H., Shankaran, K., & Desai, H. G. (1994). Wilson’s
disease: Varied hepatic presentations. Indian Journal of Gas-
troenterology, 13, 95–98. 30. Dhawan, A., Taylor, R. M., Cheeseman, P., De Silva, P., Kat-
siyiannakis, L., & Mieli-Vergani, G. (2005). Wilson’s disease in
children: 37-year experience and revised King’s score for liver
transplantation. Liver Transplantation, 11, 441–448. 12. Ren, M. S., Zhang, Z., Wu, J. X., Li, F., Xue, B. C., & Yang, R. M. (1998). Comparison of long lasting therapeutic effects
between succimer and penicillamine on hepatolenticular degen-
eration. World Journal of Gastroenterology, 4, 530–532. 31. Askari, F. K., Greenson, J., Dick, R. D., Johnson, V. D., &
Brewer, G. J. (2003). Treatment of Wilson’s disease with zinc. XVIII. Initial treatment of the hepatic decompensation presen-
tation with trientine and zinc. Journal of Laboratory and Clinical
Medicine, 142, 385–390. 13. Brewer, G. References 1. Sini, M., Sorbello, O., Sanna, F., Battolu, F., Civolani, A., Fanni,
D., et al. (2013). Histologic evolution and long-term outcome of 123 123 Combination Therapy Using Chelating Agent and Zinc for Wilson’s Disease 707 Wilson’s disease: Results of a single-center experience. Euro-
pean
Journal
of
Gastroenterology
and
Hepatology,
25,
111–117. 19. Ferenci, P., Czlonkowska, A., Stremmel, W., Houwen, R.,
Rosenberg, W., & Schilsky, M. (2012). European Association for
the Study of the Liver (EASL). Clinical Practice Guidelines:
Wilson’s disease. Journal of Hepatology, 56, 671–685. 2. El-Karaksy, H., Fahmy, M., El-Raziky, M. S., El-Hawary, M., El-
Sayed, R., El-Koofy, N., et al. (2011). A clinical study of Wilson’s
disease: The experience of a single Egyptian Paediatric Hepatology
Unit. Arab Journal of Gastroenterology, 12, 125–130. 20. Roberts, E. A., & Schilsky, M. L. (2008). American Association
for Study of Liver Diseases (AASLD). Diagnosis and treatment
of Wilson disease: An update. Hepatology, 47, 2089–2111. 3. Noureen, N., & Rana, M. T. (2011). Neurological Wilson disease
in children: A three years experience from Multan. Journal of the
Pakistan Medical Association, 61, 743–748. 21. Youn, J., Kim, J. S., Kim, H. T., Lee, J. Y., Lee, P. H., Ki, C. S.,
& Cho, J. W. (2012). Characteristics of neurological Wilson’s
disease without Kayser–Fleischer ring. Journal of the Neuro-
logical Sciences, 323, 183–186. 4. Abdel Ghaffar, T. Y., Elsayed, S. M., Elnaghy, S., Shadeed, A.,
Elsobky, E. S., & Schmidt, H. (2011). Phenotypic and genetic
characterization of a cohort of pediatric Wilson disease patients. BMC Pediatrics, 11(1), 56. 22. Walshe, J. M. (1956). Penicillamine, a new oral therapy for
Wilson’s disease. American Journal of Medicine, 21, 487–495. 23. Walshe, J. M. (1969). Management of penicillamine nephropathy
in Wilson’s disease: A new chelating agent. Lancet, 2, 1401–1402. 5. Sinha, S., Taly, A. B., Prashanth, L. K., Ravishankar, S., Arun-
odaya, G. R., & Vasudev, M. K. (2007). Sequential MRI changes
in Wilson’s disease with de-coppering therapy: A study of 50
patients. British Journal of Radiology, 80, 744–749. 24. Walshe, J. M. (2005). Hepatic Wilson’s disease: Initial treatment
and long-term management. Current Treatment Options in Gas-
troenterology, 8, 467–472. 25. Hoogenraad, T. U., Van Hattum, J., & Van den Hamer, C. J. (1987). Management of Wilson’s disease with zinc sulphate:
Experience in a series of 27 patients. Journal of the Neurological
Sciences, 77, 137–146. 6. Li, M., Zhang, Y. H., & Qin, J. (2003). References J., Askari, F., Lorincz, M. T., Carlson, M., Schilsky,
M., Kluin, K. J., et al. (2006). Treatment of Wilson disease with
ammonium
tetrathiomolybdate:
IV. Comparison
of
tetrathiomolybdate and trientine in a double-blind study of
treatment of the neurologic presentation of Wilson disease. Archives of Neurology, 63, 521–527. 32. Santos Silva, E. E., Sarles, J., Buts, J. P., & Sokal, E. M. (1996). Successful medical treatment of severely decompensated Wilson
disease. The Journal of Pediatrics, 128, 285–287. f
gy
14. Arnon, R., Calderon, J. F., Schilsky, M., Emre, S., & Shneider, B. L. (2007). Wilson disease in children: Serum aminotransferases
and urinary copper on triethylene tetramine dihydrochloride
(trientine) treatment. Journal of Pediatric Gastroenterology and
Nutrition, 44, 596–602. 33. Brewer, G. J., Yuzbasiyan-Gurkan, V., Johnson, V., Dick, R. D.,
& Wang, Y. (1993). Treatment of Wilson’s disease with zinc: XI. Interaction with other anticopper agents. Journal of the American
College of Nutrition, 12, 26–30. 34. Yonetani, L., & Walshe, J. M. (2001). Surviving Wilson’s dis-
ease. Clinical Medicine, 1, 72–74. 15. Bruha, R., Marecek, Z., Pospisilova, L., Nevsimalova, S., Vitek,
L., Martasek, P., et al. (2011). Long-term follow-up of Wilson
disease: Natural history, treatment, mutations analysis and phe-
notypic correlation. Liver International, 31, 83–91. 35. National Center for Biotechnology Information, U.S. National
Library
of
Medicine,
USA. http://www.ncbi.nlm.nih.gov/
PubMed/. Database PubMed, 2014. 16. Weiss, K. H., Gotthardt, D. N., Klemm, D., Merle, U., Ferenci-
Foerster, D., Schaefer, M., et al. (2011). Zinc monotherapy is not
as effective as chelating agents in treatment of Wilson disease. Gastroenterology, 140, 1189–1198. 36. Ferenci, P., Caca, K., Loudianos, G., Mieli-Vergani, G., Tanner,
S., Sternlieb, I., et al. (2003). Diagnosis and phenotypic classi-
fication of Wilson disease. Liver International, 23, 139–142. 17. Ranucci, G., Di Dato, F., Spagnuolo, M. I., Vajro, P., & Iorio, R. (2014). Zinc monotherapy is effective in Wilson’s disease
patients with mild liver disease diagnosed in childhood: A ret-
rospective study. Orphanet Journal of Rare Diseases, 9, 41. 37. Weiss, K. H., Thurik, F., Gotthardt, D. N., Scha¨fer, M., Teufel,
U., Wiegand, F., et al. (2013). Efficacy and safety of oral
chelators in treatment of patients with Wilson disease. Clinical
Gastroenterology and Hepatology, 11, 1028–1035. 38. Pellecchia, M. T., Criscuolo, C., Longo, K., Campanella, G.,
Filla, A., & Barone, P. (2003). Clinical presentation and 18. Frydman, M. (1990). Genetic aspects of Wilson’s disease. Jour-
nal of Gastroenterology and Hepatology, 5, 483–490. References 12 3 3 708 J.-C. Chen et al. treatment of Wilson’s disease: A single-centre experience. European Neurology, 50, 48–52. with Wilson’s disease (hepatolenticular degeneration). American
Journal of the Medical Sciences, 254, 13–23. 43. Kodama, H., Murata, Y., Iitsuka, T., & Abe, T. (1997). Meta-
bolism of administered triethylene tetramine dihydrochloride in
humans. Life Sciences, 61, 899–907. 39. Kuchinskas, E. J., & Rosen, Y. (1962). Metal chelates of DL-
penicillamine. Archives of Biochemistry and Biophysics, 97, 370. 40. Friedman, L. S., & Yarze, J. C. (1993). Zinc in the treatment of
Wilson’s
disease:
How
it
works. Gastroenterology,
104,
1566–1568. 44. Cossack, Z. T., & Bouquet, J. (1986). The treatment of Wilson’s
disease in paediatrics: Oral zinc therapy versus penicillamine. Acta Pharmacologica et Toxicologica, 59, 514–517. 41. Dubois, R. S., Rodgerson, D. O., & Hambidge, K. M. (1990). Treatment
of
Wilson’s
disease
with
triethylene
tetramine
hydrochloride (Trientine). Journal of Pediatric Gastroenterology
and Nutrition, 10, 77–81. 45. Fieten, H., Dirksen, K., van den Ingh, T. S., Winter, E. A.,
Watson, A. L., Leegwater, P. A., & Rothuizen, J. (2013). d-penicillamine treatment of copper-associated hepatitis in Lab-
rador retrievers. The Veterinary Journal, 196, 522–527. 42. McCall, J. T., Goldstein, N. P., Randall, R. V., & Gross, J. B. (1967). Comparative metabolism of copper and zinc in patients 123 123
|
https://openalex.org/W4307868866
|
https://revistadechimie.ro/pdf/9 POONAM 4 22.pdf
|
English
| null |
Review on Molecular Cross-talk of Biofilm Producing Mechanisms of Staphylococcus aureus
|
Revista de chimie
| 2,022
|
cc-by
| 5,712
|
Keywords: biofilm formation, ica locus, molecular cross-talk, MSCRAMMs, PIA, PNAG, S. aureus Keywords: biofilm formation, ica locus, molecular cross-talk, MSCRAMMs, PIA, PNAG, S. aureus Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 Review on Molecular Cross-talk of Biofilm Producing
Mechanisms of Staphylococcus aureus POONAM VERMA1*, KRISHAN KUMAR2, MANISH KUMAR VERMA3, ARUNA DUBEY2
1Department of Biotechnology, IFTM University, Delhi Road, NH-24 Moradabad, Lodhipur Rajput, Uttar Pradesh, 244102,
India
2 Department of Medicine, United Institute of Medical Sciences, Prayagraj, Uttar Pradesh, 211012, India
3 Department of Medical Biochemistry, Prasad Institute of Medical Sciences, Kanpur- Lucknow Rd, Lucknow, Uttar Pradesh,
226401, India 2 Department of Medicine, United Institute of Medical Sciences, Prayagraj, Uttar Pradesh, 211012, India
3 Department of Medical Biochemistry, Prasad Institute of Medical Sciences, Kanpur- Lucknow Rd, Luck
226401, India Abstract: Here the review converses the "molecular cross-talk" of biofuel production mechanisms for
Staphylococcus aureus. Staphylococcus aureus is a leading cause of bacterial infections globally in both
healthcare and community settings. The succes of this bacterium is the of an expansive repertoire of
virulence factors in combination with acquired antibiotic resistance and propensity for biofilm
formation. S. aureus leverages these factors to adapt to and subvert the host immune response. With the
burgeoning fiels of immunometabolism, is has become clear that the metabolic program of leukocytes
dictates their inflammatory status and overall effectiveness is clearing an infection. The treatment of S. aureus infections become complicated due to the capacity of S. aureus “multidrug-resistant” occurs
because of biofilm formationon the surfaces depending on biotic and abiotic factors, genetic factors,
and numerous environmental, which vary from species to species. A broad range of molecular
phenomenon contributes a high range of recalcitrance that is insisting on the biofilm formation. The
previous published literature illustrated that all strains of Staphylococcal sp. contain the “ica locus”
and several can form biofilms in vitro condition. Absences of “ica locus” results diminish of capability
to produce biofuels, along with "PIA gene", or mediate "N-acetyl glucosaminyl transferase activity” in
vitro condition. *email: poonam.phdbiotech@gmail.com Introduction Earlier in 1880 and 1882, Ogston confers for staphylococcal disease and its role in sepsis and abscess
formation [1]. Beyond 100 years, Staphylococcus aureus still relics as a versatile organism causing
nosocomial infection and as hazardous pathogen for humans. The staphylococcal infections amplified
progressively in community and hospital, thus responsible for mortality [2]. Staphylococci tolerate dry
condition and high salt concentrations. The staphylococci are divided into two categories, i.e. coagulase-
negative and coagulase-positive depending on the production of coagulate enzyme. MSCRAMMs
(Microbial surface components recognizing adhesive matrix molecules) may utilize by the bacteria to
get attached to human’s cell-membrane and thereby escape from immunoglobin, discovered by the
human immune defense. MSCRAMMs can also mediate adhesion to human membrane plastics and
other medical devices. Bap gene has also been isolated from S. aureus strain and has found to participate in adhering to any
surfaces as well as biofilm development phenomenon [3]. The first step of S. aureus pathogenesis is
attachment and colonization (Figure 1). The Biofilm make bacteria to get resist organized high
antimicrobial agents, concentration, environmental conditions and the host immune responses [4]. https://orcid.org/0000-0002-5688-5800 *email: poonam.phdbiotech@gmail.com Rev. Chim., 73 (4), 2022, 76-85 https://doi.org/10.37358/RC.22.4.8556 76 https://doi.org/10.37358/Rev.Chim.1949 Figure 1. Illustrate model of bacterial biofilm formation [4] Figure 1. Illustrate model of bacterial biofilm formation [4] Updated research on bacterial biofilm formation and the mechanisms Valle et al told that SarA stimulate biofilm formed by both enhancing the ica operon transcription
(Figure 2) and suppressing the transcription of proteins implicated in the proceeds of PIA/PNAG or
repressing its synthesis, whose expression would be B-dependent [5] (Figure 3). Figure 2. Schematic illustration of SarA effect on expression of independent intercellular Figure 2. Schematic illustration of SarA effect on expression of independent intercellula O’ Gara [6] demonstrated that the role of icaADBC-encoded PIA or PNAG in Staphylococcal biofilm
development, that had open to understand the pathogenicity of device-related bacterial infections. The
Teichoic acid is a copolymers component of Gram-positive bacteria such as S. epidermidis biofilm
matrix and therefore the major cyto-membrane autolysin plays a vital role within the primary
attachment section of biofilm production, whereas the cell surface biofilm-associated macro-
molecule and accumulation-associated macromolecule square measure capable of mediating alive
obsession accumulation. These findings raised, the exciting prospect that alternative surface proteins
and performance is as matter determinants or binding to living thing matrix proteins, might also act
as biofilm adhesions. Rev. Chim., 73 (4), 2022, 76-85 https://doi.org/10.37358/RC.22.4.8556 77 Revista de Chimie https://revistadechimie.ro https://doi.org/10.37358/Rev.Chim.1949 Figure 3. Genomic representation of the ica operon and surrounding sequence in S. aureus Figure 3. Genomic representation of the ica operon and surrounding sequence in S. au BC dependent biofilm forming pathway in recombinant strains of S. aureus Arana et al. evaluated that Hussain-Hastings-White modified [HHWm] medium determined the
factors that reduce the formation of biofilm process by mutagenesis and S. aureus biofilm strain
systematic disruption. The arlRS mutant showed the PNAG participated in primary attachment and
gathering process in biofilm formation. Biofilm formation is done with the help of arlRS mutant that
unable to show any changes [7]. Bap surface protein occupied, while biofilm development in S. aureus strain, that is isolated from
chronic mastitis infections. In this study, Bap orthologue genes were isolated from many coccus species,
i.e, S. chromogenes, S. simulans, S. xylosus, S. epidermidis, and S. hyicus. However, flanking region
sequence analyses discovered that the Bap factor of those species wasn't contained within the SaPI bov2
pathogenicity island. Even though they didn't contain icaADBC deoxyribonucleic acid, all the coagulase-
negative coccus isolates harboring Bap, were robust biofilm producers. Annoyance of the Bap factor in
S. epidermidis eradicated its ability to provide biofilm, while the heterologous complementation’s of
biofilm-negative strain belongs to S. Rev. Chim., 73 (4), 2022, 76-85 Updated research on bacterial biofilm formation and the mechanisms Chim., 73 (4), 2022, 76-85 https://doi.org/10.37358/RC.22.4.8556 78 Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 combined result of mutating nuc and adding enzyme inhibitors leading to the formation of biofilm by
means of SarA mutant, approached that of UAMS-1 parent strain. combined result of mutating nuc and adding enzyme inhibitors leading to the formation of biofilm by
means of SarA mutant, approached that of UAMS-1 parent strain. Memmi et al. considered that lysis plays a necessary role in the microorganism biological process
and lactam antibiotics [12]. Accordingly, the autolysins expression is firmly regulated by various
endogenous regulators, arlRS gene, by two part restrictive systems that showed the negatively regulated
lysis in MSSA strains that is masculine-sensitive. The study evidenced that the inactivation of "arlRS"
does not lysis the MRSA strains that were methicillin-resistant. In contrast with MSSA strains, Newman,
SH1000, RN6390, and 8325-4, lysis was plagued by arlRS gene. Memmi again determined that the
hanging characteristic function of arlRS gene between MSSA and MRSA strains are not because of the
penicillin resistance determinant mecA gene. Table 1. Comparison between MRSA and MSSA strains regarding biofilm formation
S. No
MRSA strain
MSSA strain
1
Biofilm formation in the main factor controlling MRSA is
surface adherence. which repressed under agr expression [15]
In MSSA, the creation of biofilms is more reliant
on cell-to-cell adhesion by icaADBC-encoded
PIA/PNAG or slime [15]
2
Multilocus sequence typing using five key genes (MLST)
clonal complexes (CCs), i.e. CC5, CC8, CC22, CC30 and
CC45 contributes to the biofilm formation under physiologic
glucose concentration [16]
(MLST) clonal complexes (CCs), i.e. Updated research on bacterial biofilm formation and the mechanisms CC1
contribute to the biofilm formation [17]
3
MRSA strain shows dry crystalline s(rough) morphology (slime
producing positive), near about 0% [15]
MSSA strains show a deviant, dry crystalline
(rough) morphology (slime producing positive),
near about 14% [15]
4
MRSA strain was tested positive (mecA+) for the MRSA-
specific mecA gene, by real-time multiplex PCR [18]
MSSA strain was tested negative ((mecA–)
for the mecA gene
5
MRSA biofilm growth involves protein adhesions controlled by
SarA and Agr and is ica independent [15]
SarA-regulated PIA/PNAG plays major in MSSA
biofilm development [15]
6
NaCl activated biofilm development shows the minor biofilm
matrix in MRSA isolates responded to NaCl [15]
NaCl activated biofilm development show the
major biofilm matrix in MSSA isolates responded
to NaCl [15]
7
1 percent glucose supplement was added to BHI medium, the
biofilm matrix is minor on MRSA clinical isolates [15]
1 percent glucose supplement was added to BHI
medium, the biofilm matrix is major on MSSA
clinical isolates [15] aADBC independent biofilm forming pathway in recombinant strains of S. aureus icaADBC independent biofilm forming pathway in recombinant strains of S. aureus Fitzpatrick et al. investigated that clinical isolates of staphylococcal species, icaADBC-encoded PIA
or PNAG enzymes play a vital role in biofilm formation mechanism [17]. By clinical isolates of MRSA
strain, environmental factors don't continually contribute for the increment of the biofilm formation
method, but in clinical isolates of MRSA due to addition of aldohexose thus shows icaADBC freelance
pathway. According to Fitzpatrick, ica operon are not necessary for the formation of biofilm on BHI
media, however it revealed that the ica locus was necessary for biofilm formation in clinical isolates of
S. aureus. In this study, Fitzpatrick concluded the control of biofilm phenotype phenomenon for clinical
isolates staphylococci species by regulatory mechanisms. Boles and Horswill [18] revealed that the agr protein was the essential factor of Staphylococcus
aureus that participated in quorum-sensing system and icaADBC mediated biofilm formation pathways. Recent study discusses about the role of the agr mutants in agr system of S. aureus biofilm formation,
cells dispersing from biofilm have been showing an active agr system. Boles and Horswill discussed the
involvement of S. aureus bacterium in the formation of biofilms via both mechanisms that was ica
dependent and/or ica-independent. O’Neill et al. evaluated the biofilm development in MRSA follows the icaADBC independent
pathway [19]. Updated research on bacterial biofilm formation and the mechanisms aureus with the Bap super-molecule from S. epidermidis bestowed
the capability to make a biofilm on a phenyl ethylene surface. In general, the outcomes reveal that Bap
orthologues from “coagulase-negative” Staphylococci trigger a different mechanism for the formation
of biofilm, independent of “PIA/PNAG exopolysaccharide” [8]. SarA mutants unable to synthesize Bap-dependent biofilm due to an agr-independent mechanism. However, Bap promoter characterized, through applying rapid amplification of c-DNA ends technique
the transcriptional start point was mapped. Trotonda et al demonstrated that SarA signified the regulation
of the biofilm formation process within S. aureus strain. This study proved that in present of SarA clone
biofilm development was decreased in presence of SarA mutation [9]. Sambanthamoorthy et al. explored the S. aureus as an important nosocomial bacterium that have
competence for biofilm formation because of SarA gene. SarA gene plays vital part in the formation of
biofilms process and further known as msa gene. It was necessary for controlled the virulence factors
and showed SarA expression. Here msa gene decreased the expression of SarA gene for biofilm
development and mutant of msa formed a weak and unstable biofilm whereas, also analysed that
bacterial biofilm formation mechanism at the molecular level is complex process and environmental
factors and some independent regulators played significant part in the development of assembly of S. aureus strain on nature. Sambanthamoorthy also concluded that msa gene is a necessary protein in
biofilm formation process like SarA gene [10]. Tsang et al. shown that a mutant of SarA decreased the biofilm forming capacity, however on the
basis of mechanism, the functions are not known yet and also reported that identical genes had
participated in biofilm formation [11]. The SarA gene engaged in synthesis of nucleolytic and proteolytic
exoenzymes, acid tolerance. The inability of mutant repeatedly expresses independent production of
extracellular nuclease and multiple proteases, however, gathered effects contributes in the biofilm -
deficient phenotype of SarA mutated S. aureus. This study suggests that the reduced capability of SarA
mutant creates a biofilm, involves proteases that are serine, amino acid and metalloproteases. Inclusion
of enzyme inhibitors conjointly increased biofilm formation in a very SarA/nuc mutant, with the Rev. Updated research on bacterial biofilm formation and the mechanisms In MRSA, biofilm development is encouraged by acidic growth medium condition and Rev. Chim., 73 (4), 2022, 76-85 https://doi.org/10.37358/RC.22.4.8556 79 Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 triggered by adding glucose into the growth medium. If mutations incorpurated in fnbA and fnbB proteins
of MRSA, they reduced the biofilm formation process; however, these mutants have no effect on the
MSSA biofilm formation. FnBP had not confirmed any relationship in the primary attachment of
biofilm, however, encouraged at intercellular accumulation level. FnBP furthermore encouraged the
biofilm formation that’s completely dependent on SarA besides this does not show any effect on fnbA or
fnbB transcription. S. aureus biofilm formation gets optimized with fnbA and fnbB proteins, showed
their independent pathway for known ligand binding activities of the multifunctional surface proteins. The study illustrated that extracellular matrix binding proteins fnbA and fnbB of S. aureus participated
in intracellular accumulation and biofilm formation. The implanted surgical and medical biomaterials
are coated through a film. These films contain extracellular matrix proteins like- fibronectin. In MRSA
virulence factors are cell wall- anchored proteins. Houston et al. determined the atl protein role in the formation of ica-independent biofilm pathway
within S. aureus [20]. The studies evidenced the role of actual protein for biofuel production on
hydrophilic polystyrene substance and therefore provided an excellent surface for cell attachment and
growth. In this study, they determined the role of sigma factor sigB that decreased the extracellular
protease production and RNAIII expression with the help of FnBP. A sigma factor didn't play any role
in PIA-dependent biofilm development. The Mutant agr locus macromolecule participated to increase
the FnBP-dependent biofilm formation, whether the SarA mutation, that encourage the production of
proteinase and closed to their biofilm development function that was mediated by FnBP. For a second
time Houston analyzed the regulation of atl gene, every time atlR enhanced the autolysin process and
atlR::Tcr mutation in BH1CC increased biofilm-forming capacity. Throughout the study, Houston
accomplished that atl macromolecule is essential for the early stage of "FnBP-dependent" S. aureus
biofilm phenotype for autolysin. Here Houston accomplished the role of atlR protein, agr protein, SarA
protein and sigB proteins for biofilm formation in S. aureus and involvement of atl protein in initial
attachment and release of eDNA at initial stages of biofilm development via ica-independent and FnBP-
mediated process. Updated research on bacterial biofilm formation and the mechanisms aureus
and determine the development of biofilm via Atl-mediated lysis. aureus. The study makes insight relative contributions of the above studied both proteins in S. aureus
and determine the development of biofilm via Atl-mediated lysis. Pozzil et al. illustrated that the biofilm phenotypes were supported by autolysin of cell wall and a
binding protein called fibronectin as well as the icaADBC-encoded PIA/PNAG were also illustrated
from clinical isolated of S. aureus [23]. The process of Biofilm formation in MSSA strain was dependent
on PIA/PNAG however, in isolates that were MRSA express an atl/FnBP-mediated biofilm phenotype
reveled correlation between process of bioflm synthesis and susceptibility for β-lactam antibiotics. The
S. aureus causes nosocomial infections that reported universally and express function i.e. resistance
organized antibiotics, production of enzymes and toxin, biofilm forming and capacity of immune
evasion. Puzzle explained that MSSA was more preferred to form PNAG-dependent biofilm rather than
MRSA isolates that produce biofuels, which was atl/FnBP-dependent and thus explained the roles of the
methicillin susceptibility affects the expression of biofilm. Updated research on bacterial biofilm formation and the mechanisms The atl act as primary attachment for substance along with the assistance of
macromolecule lysis through the cell lysis, eDNA release, and initial cell accumulation during the
development of a biofilm and while maturation, "FnBPs" played an essential role. Lei et al. determined that the biofilm formation method in S. aureus MW2 strain behaved as a
virulence factor [21]. The development of Biofilms may be a complex process that includes
polysaccharide, protein, and elements of DNA that was maintained by various control factors. In S. aureus MW2 strain, Rsp repress the steps involve in biofilm formation, thus reveal attachment and
biofilm formation process through the gene fnbA. The conclusion of the study illustrated that S. aureus
formation of biofilms and their regulations, both were very complicated method because of association
of multiple elements throughout development that was enclosed to the sugar, proteins, and extracellular
DNA. All through this work, Lei accomplished that the family regulaters involved AraC/XylS factors,
whereas Rsp sequence inhibits the biofilm formation in S. aureus MW2 strain. p
q
Bose et al. said that the most specific murrain hydrolase of S. aureus encoded by AtlA gene and by
proteolytic cleavage of bifunctional enzyme, two catalytically active proteins were obtained, i.e. amidase
(AM) and glucosaminidase (GL) [22]. Most studies summarize the combined functions of the proteins
for metabolic activity of cell wall and biofilm formation. Through this study, Bose discovered derivatives
of mutant for clinical isolate of S. aureus strain and UAMS-1, where single or even both AM and GL
domains of AtlA gene have been deleted. After these strains were studied, it was discovered that each
mutant had growth rates similar to those of the parental strain. However, express clumping phenotypes
and lysis profiles distinct from the parents and offspring strain thus suggesting the distinct roles in cell
wall metabolism. Bose analyzed the activity of the mutants for biofilm assays and found that both
proteins were dominant for biofilm development, together with the function of analysis that release
genomic DNA for synthesizing biofilm matrix molecules. Moreover, the application of enzymatically
inactive point mutations uncovered the catalyst activity of both the proteins in biofilm formation in S. Rev. Chim., 73 (4), 2022, 76-85 https://doi.org/10.37358/RC.22.4.8556 80 Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 aureus. The study makes insight relative contributions of the above studied both proteins in S. Interactions between icaADBC-dependent and -independent proteins biofilm forming pathways
in recombinant strains of S. aureus In mouse, determined chronic infection was caused due to bap protein. In S. aureus stain,
ica locus formed the PIA-PNSG for biofilm development. In this result, Cucarella concluded the
correlation between BAP-PIA-PNSG and found that both strains bap+ and bap- produced the PIA-PNSG
enzyme. Cucarella discovered the new specific types of proteins from S. aureus that is known to us as
bap, which was involved in biofilm formation and grow on artificial medium. These proteins participated
in attachment of both types of species. yp
p
Gross et al. determined the role of clinical isolates of S. aureus played for attachment to artificial
surface, i.e. implanted biomedical devices [26]. However, the mechanism for primary attachment to bio-
medical devices is unknown. Gross accomplished that electrostatic forces reduce the process of biofilm
development in clinical isolates of staphylococcal species and teichoic acids involve pre-dominantly in
the initial step of biofilm synthesis and/or colonization on medical devices. Cucarella et al. carried out a study over a period of 3 months at Cardenal Herrera-CEU University,
Spain and accomplished that S. aureus is a frequent reason of nosocomial infectious and intramammary
infectious diseases in human-being and bovine animals that often become chronic and allied with the
capacity to produce biofilm by bacteria [25]. Here, cucarella illustrate a correlation to produce chronic
bovine mastitis and the formation of biofilm. Cucarella divided the bacteria “S. aureus (bovine mastitis)”
into 3 groups, based on their genetic elements. The group 1 includes ica+ bap+, group 2 includes ica+,
bap-, and group 3 includes ica_, bap_ respectively. Cucarella discovered that bap gene were identified
on the basis of structure. Cucarella accomplished that intramammary gland presents in a bovine body
that occupied a significant role in the biofilm development in S. aureus. The study concluded that bovine
mastitis disease emerged due to biofilm formation in S. aureus strains and bap is the most imperative
gene for that phenomenon. Resch et al. screened that the bacteria synthesize biofilm confirmed the extreme resistivity against
compared to their planktonic counterparts, antibiotics and the immune system and elucidate that biofilm
cells showed different metabolic activity [27]. Resch implicited that staphylococci species bacteria
protected themselves from pigment formation on exposing to UV- radiation and radical’s in vivo
condition. Interactions between icaADBC-dependent and -independent proteins biofilm forming pathways
in recombinant strains of S. aureus Cramton et al. evaluated that the nosocomial infections due to hospitalization because of the
formation biofilm on the biomedical implant surfaces, causing sepsis by colonization of Staphylococcus
species [24]. Biofilm process involves 2 steps: first is cell-cell adhesion and second is the multiple layer
formation. Through this work, icaADBC locus, which is involved in the formation of extracellular
polysaccharide adhesion termed PIA or PNA enzymes and that having a linkage UDP-N-acetylgluco-
samine in vitro condition had been analyzed. The researchers placed an interesting about all the
Staphlococcus species have icaADBC locus, able to developed biofilm in-vitro condition (Figure 4). S. aureus and S. epidermidis are gram-positive cocci and able attach with biomedical surfaces for
developing biofilm and thus concluded that both the species formed biofilm in 2 steps, i.e. cell to cell
adhesion because of ica operon and capable of PIA production and development of biofilm in-vitro
condition. The Cramton also accomplished that S. aureus bacteria caused nosocomial infections in the
human being and shows the high death and morbidity rates as well as spread high frequency of infection
by both S. aureus and S. epidermidis strains by means of ica gene. Figure 4. icaADBC-dependent and -independent pathway-associated proteins [4] Figure 4. icaADBC-dependent and -independent pathway-associated proteins [4] Cramton et al. determined that S. aureus and S. epidermidis strains formed the biofilm via operon
"intercellular adhesion" (ica) and formed a linear β-1,6-linked glucosaminylglycan [24]. For assembling
biofilm steps, cell to cell adhesions are needed in order so that biofilm method will increase the resistance
and virulence nature in both strains. Through the work, Cramton additionally concluded that icaADBC-
encoded PIA/PNAG enzymes contributed the necessary role in the biofilm formation under anaerobic
conditions. Cramton accomplished the development of the biofilm technique's molecular cross-talk,
which acts as a virulence factor in an anaerobic environment in vivo conditions, was facilitated by Rev. Chim., 73 (4), 2022, 76-85 https://doi.org/10.37358/RC.22.4.8556 81 Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 icaADBC-encoded (PIA/PNAG) enzymes. Throughout this manuscript, Cramton said that anaerobic
surroundings, conditions affected the PIA/PNSG production method in S. aureus and S. epidermidis
stains. Cucarella et al. demonstrated that bap protein has found in S. aureus surface and contribed for biofuel
development as well as recognized the involvement of new genes in biofilm development [25]. All
staphylococcus stains having bap proteins are capable to form a high adhesion power for biofilm
development. Rev. Chim., 73 (4), 2022, 76-85 Interactions between icaADBC-dependent and -independent proteins biofilm forming pathways
in recombinant strains of S. aureus According to Resch, SsaA is a staphylococcal secretary antigen that contributes in disease
related to biofilm and anti-SsaA immunoglobulin G antibody are often present in human serum of S. epidermidis strain, which was endocarditis. Research explained that S. aureus stain gene is essential for
detoxification procedure of reactive oxygen species (ROS) and thus illustrated expression at higher
levels in biofilm cells. O'Neill et al. determined that the pathway of staphylococci species was formed i.e. dependent or
independent, and form via icaADBC-encoded PIA/PNAG [19]. Here, O'Neill illustrates that methicillin
is essential for the phenotype of biofilm in S. aureus strain. O'Neill again described the roles of
icaADBC-encoded PIA/PNAG in MRSA and MSSA stains for the process of biofilm formation. O'Neill
concluded that biofilm formation in MRSA strain is due to the ica independent pathway and the pathway
is governed by SarA and agr proteins but in MSSA strain biofilm formation is governed by SarA. Tu Quoc et al. investigated that S. aureus produces the biofilm and the colonization using this mode
facilitates infections, is very complicated to treat and confirm high morbidity and mortality [28]. To
create an international mutant library, they exploited bacteriophage Mu transposition methods for highly
biofilm-forming S. aureus clinical isolate. In S. aureus and S. epidermidis both strains have the capacity
to form biofilm due to PIA and produced by icaADBC operon having insertions. The S. aureus S30
strain of clinical isolates are collected from a Geneva hospital that caused various serious diseases due
to the formation of biofilm in human-being. Disruption of Em-Mu in icaADBC demonstrated the utility Rev. Chim., 73 (4), 2022, 76-85 https://doi.org/10.37358/RC.22.4.8556 82 Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 of PIA by obtaining a high proportion of independent, for biofilm development in this clinical isolates
of S. aureus strain and express the strong validation for procedure of screening, which concomitantly
uncovered additional mutants. The strain was explored as a model for identification of genes, which
involved in major role in biofilm production. In this study, Tu Quoc concluded that Em-Mu insertions
were presented in only 2 factors i.e. fmtA and atpF. These factors didn't show any relationship among
WTA staining technique and mutants of the above factors for production of Teichoic acid wall. Schroeder et al. demonstrated that Staphylococci species are leading causes of implant-associated
infections universally, as its colonies formed on implanted medical devices [29]. References
1.OGSTON A., Micrococcus poisoning. J Anat. 1882; 17: 24-58.
2.LOWY FD., Staphylococcus aureus infections. New England J Med. 1998; 339: 520-532.
3.CUCARELLA C., SOLANO C., VALLE J., AMORENA B., LASA I., ET A.L., bap, a Staphylococcus
aureus surface protein involved in biofilm formation. J Bact. 2001; 183: 2888–2896.
4.VERMA P, MAHESHWARI SK, MATHUR A. A review on Bacterial Biofilm Formation and
Disassembly. Int J Pharm Sci Res. 2013; 4(8): 2900-2906. Interactions between icaADBC-dependent and -independent proteins biofilm forming pathways
in recombinant strains of S. aureus Bacteria attach to the
surface of a medical device to multiply and build up into multilayered cell clusters known as biofilms. The biofilm formations were also mediated by carbohydrate and macromolecule. It was accomplished
that S. aureus have "specific surface protein factor (SasC)" that participated in aggregation of cell, same
colonization-related biofilm formation and facilitate the accumulation in infected bacterium. Through
this study, Schroeder accomplished that nosocomial isolated organisms are associated with biofilm
forming process with artificial surfaces, owing S. aureus and "coagulase-negative" staphylococci
illustrating the high rates of morbidity and mortality. Gruszka et al. observed the both S. aureus and S. epidermidis are capable to formed an assembly like
structure, called biofilms artificial surfaces and thus cause the infections that affect worldwide peoples
and cause morbidity as well as mortality [30]. Because of resistance against antibiotics and device
removal, biofilms are commonly needed to resolve the infection. Thus, they were needed to grow new
therapies and molecular information, to assist. Accumulation-associated supermolecule (Aap) and
Surface proteins (SasG) promote the biofilm formation of S. epidermidis through "B" regions. “B”
regions contain tandem array of “G5 domains" about fifty residue sequences (referred to as E here) are
dotted throughout, and it has been suggested that these sequences may act as a mediator of intercellular
accumulation via Zinc2+-mediated homodimerization. Although unstructured E areas are predicted,
SasG and Aap form extensive fibrils on the surface of the bacterium. Conclusions The Gram-positive S. aureus nosocomial pathogen that is connected to infection related to medical
implants. The stage of cell to cell adhesion is mediated by the bacterial species in biofilm development
process and considered as a complicated phenomenon. Around 80% diseases had caused due to biofilm
formation on medical devices. The review article established that some mutant i.e. SarA, ica, agr, fnbA,
fnbB, arlRS, sigB, and sarZ etc participating in adhesion to any surfaces and biofilm forming
Staphylococcus bacterial species. The ica locus is a crucial target of potential therapy for the avoidance
of persistent infections linked to prosthetic medical devices because due to the substantial morbidity and
mortality brought on by S. aureus infections in addition to the rate of infection via both species. A more
complete understanding in molecular cross-talk of biofilm producing mechanisms will lead the
development of novel therapies to understand biofilm formation. Future prospects: Biofilms are understood as bacterial communities that adhere to surfaces by
encasing into a self-produced extracellular polymeric matrix. The staphylococcal biofilm matrix may
contain exopolysaccharides and proteins as well as extracellular (e)DNA. A more complete
understanding in molecular cross-talk of biofilm producing mechanisms will lead the development of
novel therapies to understand biofilm formation. References https://doi.org/10.37358/RC.22.4.8556 Rev. Chim., 73 (4), 2022, 76-85 83 Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 Revista de Chimie OTTO M., Staphylococcal biofilms. Curr Top Microbiol Immunol. 2008; 322: 207-228. 5.OTTO M., Staphylococcal biofilms. Curr Top Microbiol Immunol. 2008; 322: 207-228. 6.ZHANG Y.Q., REN S.X., LI H.L., et al. Genome-based analysis of virulence genes in a non-biofilm-
forming staphylococcus epidermidis strain (ATCC 12228). Mol Microbiol. 2003; 49: 1577-1593. 7.VALLE J., ARANA T.A., BERASAIN C., GHIGO J.M., AMORENA B., et al. SarA and not sigma B
is essential for biofilm development by Staphylococcus aureus. Mol Microbiol. 2003; 48: 1075–1087. 8.O'GARA J.P., ica and beyond: Biofilm mechanisms and regulation in staphylococcus epidermidis and
staphylococcus aureus. FEMS microbiol lett. 2007; 270: 179-188. 6.ZHANG Y.Q., REN S.X., LI H.L., et al. Genome-based analysis of virulence genes in a non-biofilm-
forming staphylococcus epidermidis strain (ATCC 12228). Mol Microbiol. 2003; 49: 1577-1593. 7.VALLE J., ARANA T.A., BERASAIN C., GHIGO J.M., AMORENA B., et al. SarA and not sigma B
is essential for biofilm development by Staphylococcus aureus. Mol Microbiol. 2003; 48: 1075–1087. 7.VALLE J., ARANA T.A., BERASAIN C., GHIGO J.M., AMORENA B., et al. SarA and not sigma B
is essential for biofilm development by Staphylococcus aureus. Mol Microbiol. 2003; 48: 1075–1087. 8.O'GARA J.P., ica and beyond: Biofilm mechanisms and regulation in staphylococcus epidermidis and
staphylococcus aureus. FEMS microbiol lett. 2007; 270: 179-188. is essential for biofilm development by Staphylococcus aureus. Mol Microbiol. 2003; 48: 1075 1087. 8.O'GARA J.P., ica and beyond: Biofilm mechanisms and regulation in staphylococcus epidermidis and
staphylococcus aureus. FEMS microbiol lett. 2007; 270: 179-188. 9.ARANA A.T., MERINO N., IRIGARAY M.V., DEBARBOUILLE M., PENADE´S J.R., et al. Staphylococcus aureus develops an alternative, ica-Independent biofilm in the absence of the arlRS
Two-Component System. J Bact. 2005; 187: 5318–5329. p
y
10.TORMO M.A., KNECHT E., GOTZ F., LASA I., PENADES J.R., Bap-dependent biofilm formation
by pathogenic species of Staphylococcus: evidence of horizontal gene transfer? Microbiol. 2005; 151:
2465–2475. 11.TROTONDA M.P., MANNA A.C., CHEUNG A.L., LASA I., PENADE J.R., SarA Positively
controls bap-dependent biofilm formation in Staphylococcus aureus. J Bact. 2005; 187: 5790–5798. 12.SAMBANTHAMOORTHY K., SCHWARTZ A., NAGARAJAN V., ELASRI M.O., The Role of
msa in Staphylococcus aureus biofilm formation. BMC Microbiol. 2008; 8: 01-09. 13.TSANG L.H., CASSAT J.E., SHAW L.N., BEENKEN K.E., SMELTZER M.S., Factors contributing
to the Biofilm-deficient phenotype of Staphylococcus aureus sarA Mutants. PLoS One. 2008; 3: 01-14. References 14.MEMMI G., NAIR D.R., CHEUNG A., Role of ArlRS in Autolysis in Methicillin-Sensitive and
Methicillin- Resistant Staphylococcus aureus Strains. J Bacteriol. 2011; p:759–767. p y
p
15.O’NEILL E., POZZI C., HOUSTON P., SMYTH D., HUMPHREYS H., ROBINSON D.A.,
O’GARA J.P., Association between methicillin susceptibility and biofilm regulation in staphylococcus
aureus isolates from device-related infections. J Clin Microbiol. 2007; 45(5): 1379-1388. 16.DEURENBERG R.H., VINK C., KALENIC S., FRIEDRICH A.W., BRUGGEMAN C.A.,
STOBBERINGH E.E., The molecular evolution of methicillin-resistant Staphylococcus aureus. Clin
Microbiol Infect. 2007; 13(3): 222-235. 17.NOTO M.J., KREISWIRTH B.N., MONK A.B., ARCHER G.L., Gene acquisition at the insertion
site for SCCmec, the genomic island conferring methicillin resistance in Staphylococcus aureus. J
Bacteriol. 2008; 190(4): 1276-1283. 18.DONKER G.A., DEURENBERG R.H., DRIESSEN C., SEBASTIAN S., NYS S., STOBBERINGH
E.E., The population structure of Staphylococcus aureus among general practice patients from The
Netherlands. Clin Microbiol Infect. 2009; 15(2): 137-143. 19.FITZPATRICK F., HUMPHREYS H., O’GARA J.P., Evidence for icaADBC-independent biofilm
development mechanism in methicillin-resistant Staphylococcus aureus clinical isolates. J Clin
Microbiol. 2005; 43: 1973–1976. 20.BOLES B.R., HORSWILL A.R., agr-mediated dispersal of Staphylococcus aureus biofilms. PLoS
Pathog. 2008; 4: 01-13. 21.O’NEILL E., POZZI C., HOUSTON P., HUMPHREYS H., ROBINSON D.A., et al. A Novel
Staphylococcus aureus Biofilm Phenotype Mediated by the Fibronectin-Binding Proteins, FnBPA and
FnBPB. J Bacteriol. 2008; 190: 3835–3850. 22.HOUSTON P., ROWE S.E., POZZI C., WATERS E.M., O’GARA J.P., Essential Role for the Major
Autolysin in the Fibronectin-Binding Protein-Mediated Staphylococcus aureus Biofilm Phenotype. Infect Immun. 2011; 79: 1153–1165. f
23.LEI M.M., CUE D., ROUX C.M., DUNMAN P.M., LEE C.Y., Rsp inhibits attachment and biofilm
formation by repressing fnbA in Staphylococcus aureus. J Bacteriol. 2011; 193: 5231–5241. 24.BOSE J.L., LEHMAN M.K., FEY P.D., BAYLES K.W., Contribution of the Staphylococcus aureus
Atl AM and GL Murein Hydrolase Activities in Cell Division, Autolysis, and Biofilm Formation. PLoS
One. 2012; 7: 01-11. Rev. Chim., 73 (4), 2022, 76-85 https://doi.org/10.37358/RC.22.4.8556 84 Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 Revista de Chimie
https://revistadechimie.ro
https://doi.org/10.37358/Rev.Chim.1949 Revista de Chimie 25.POZZI C., WATERS E.M., RUDKIN J.K., SCHAEFFER C.R., LOHAN A.J., et al. Methicillin
resistance alters the biofilm Phenotype and attenuates virulence in Staphylococcus aureus device-
associated infections. PLoS Pathog. 2012; 8: 01-15. g
26.CRAMTON S.E., GERKE C., SCHNELL N.F., NICHOLS W.W., GOTZ F., The intercellular
adhesion (ica) locus is present in Staphylococcus aureus and is required for biofilm formation. Infect
Immun. 1999; 67: 5427–5433. Rev. Chim., 73 (4), 2022, 76-85 References 27.CUCARELLA C., TORMO M.A., UBEDA C., TROTONDA M.P., MONZON M., PERIS C., et al. Role of Biofilm-Associated Protein bap in the Pathogenesis of Bovine Staphylococcus aureus. Infect
Immun. 2004; 72: 2177–2185. 28.GROSS M., CRAMTON S.E., GOTZ F., PESCHEL A., Key Role of Teichoic Acid Net Charge in
Staphylococcus aureus Colonization of artificial surfaces. Infect Immun. 2001; 69: 3423–3426. 29.RESCH A., ROSENSTEIN R., NERZ C., GO¨TZ F., Differential Gene Expression Profiling of
Staphylococcus aureus cultivated under Biofilm and Planktonic Conditions. Appl Environ Microbiol. 2005; 71: 2663–2676. 30.TU QUOC P.H., GENEVAUX P., PAJUNEN M., SAVILAHTI H., GEORGOPOULOS C., et al. Isolation and characterization of biofilm formation-defective mutants of Staphylococcus aureus. Infect
Immun. 2007; 75: 1079–1088. 31.SCHROEDER K., JULARIC M., HORSBURGH S.M., HIRSCHHAUSEN N., NEUMANN C., et
al. Molecular characterization of a novel Staphylococcus aureus surface protein (Sas C) involved in cell
aggregation and biofilm accumulation, PLoS One. 2009; 4: 01-14. gg g
32.GRUSZKA D.T., WOJDYLA J.A., BINGHAM R.J., TURKENBURG J.P., MANFIELD I.W., et al. Staphylococcal biofilm-forming protein has a contiguous rod-like structure. J PNS. 2012; pp: 01-08. Manuscript received: 20.06.2022 Rev. Chim., 73 (4), 2022, 76-85 85 https://doi.org/10.37358/RC.22.4.8556
|
https://openalex.org/W4318618323
|
https://www.nature.com/articles/s41467-023-36207-7.pdf
|
English
| null |
Global vegetation resilience linked to water availability and variability
|
Nature communications
| 2,023
|
cc-by
| 9,579
|
Article https://doi.org/10.1038/s41467-023-36207-7 Global vegetation resilience linked to water
availability and variability Indeed, recent work4,5,14,15 has
shown that many regions are losing vegetation resilience; however, the
drivers of spatial heterogeneity in resilience and resilience trends
re
un
ca
sy
au
sy
ci
it
re
qu
th
pr
gl
w
ve
fit
ex
re
M
di
tu
1Institute of Geosciences, Universität Potsdam, Potsdam, Germany. 2Potsdam Instit
of Munich, School of Engineering & Design, Earth System Modelling, Munich, Germ
of Exeter, Exeter, UK. e-mail: tasmith@uni-potsdam.de
Nature Communications| (2023) 14:498 The resilience of ecosystems, i.e., their capacity to resist and recover
from external perturbations—natural or anthropogenic—has received
increasing attention in recent years1–5. Key to this discussion is whether
ecosystems can potentially exhibit multiple stable equilibrium states
with abrupt transitions between them in response to gradual changes
in climatic and environmental conditions. For some regions, it has
even been suggested that alternative stable states may co-exist for the
same climatic forcings; for example, it is thought that several tropical
regions support both a stable rainforest and a stable savanna state for a
considerable range of mean annual precipitations6–8. The capacity of
ecosystems to recover to their previous state after a shock—such as a
fire, drought, or deforestation—is a critical open question, particularly
in view of the impacts of anthropogenic climate change9. Changes in
ecosystem
function
can
drastically
alter
carbon
sequestration
capacities10; for example, the Amazon rainforest appears to have
recently turned from a globally relevant carbon sink to a net source of
carbon11. The potential for abrupt transitions between alternative
stable states—and corresponding risks of further carbon emissions—is
not only confined to the tropics12,13, making identifying controls on
ecosystem resilience a global concern. Indeed, recent work4,5,14,15 has
shown that many regions are losing vegetation resilience; however, the
drivers of spatial heterogeneity in resilience and resilience trends remain unconstrained. In this work, we therefore aim to improve our
understanding of possible climatic drivers of vegetation resilience. A wide body of previous research16,17 has proposed that the
capacity of a system to recover from external shocks, and hence the
system’s resilience, is closely tied to both the variance and the lag-one
autocorrelation (AC1) of time series encoding the dynamics of the
system in question1,18–22; higher values of AC1 and variance are asso-
ciated with lower resilience (see Methods). 1Institute of Geosciences, Universität Potsdam, Potsdam, Germany. 2Potsdam Institute for Climate Impact Research, Potsdam, Germany. 3Technical University
of Munich, School of Engineering & Design, Earth System Modelling, Munich, Germany. 4Department of Mathematics and Global Systems Institute, University
of Exeter, Exeter, UK.
e-mail: tasmith@uni-potsdam.de Global vegetation resilience linked to water
availability and variability Taylor Smith
1
& Niklas Boers
2,3,4 Taylor Smith
1
& Niklas Boers
2,3,4 Received: 5 November 2021
Accepted: 18 January 2023
Check for updates Quantifying the resilience of vegetated ecosystems is key to constraining both
present-day and future global impacts of anthropogenic climate change. Here
we apply both empirical and theoretical resilience metrics to remotely-sensed
vegetation data in order to examine the role of water availability and variability
in controlling vegetation resilience at the global scale. We find a concise global
relationship where vegetation resilience is greater in regions with higher water
availability. We also reveal that resilience is lower in regions with more pro-
nounced inter-annual precipitation variability, but find less concise relation-
ships between vegetation resilience and intra-annual precipitation variability. Our results thus imply that the resilience of vegetation responds differently
to water deficits at varying time scales. In view of projected increases in pre-
cipitation variability, our findings highlight the risk of ecosystem degradation
under ongoing climate change. The resilience of ecosystems, i.e., their capacity to resist and recover
from external perturbations—natural or anthropogenic—has received
increasing attention in recent years1–5. Key to this discussion is whether
ecosystems can potentially exhibit multiple stable equilibrium states
with abrupt transitions between them in response to gradual changes
in climatic and environmental conditions. For some regions, it has
even been suggested that alternative stable states may co-exist for the
same climatic forcings; for example, it is thought that several tropical
regions support both a stable rainforest and a stable savanna state for a
considerable range of mean annual precipitations6–8. The capacity of
ecosystems to recover to their previous state after a shock—such as a
fire, drought, or deforestation—is a critical open question, particularly
in view of the impacts of anthropogenic climate change9. Changes in
ecosystem
function
can
drastically
alter
carbon
sequestration
capacities10; for example, the Amazon rainforest appears to have
recently turned from a globally relevant carbon sink to a net source of
carbon11. The potential for abrupt transitions between alternative
stable states—and corresponding risks of further carbon emissions—is
not only confined to the tropics12,13, making identifying controls on
ecosystem resilience a global concern. Global vegetation resilience linked to water
availability and variability Note that higher values correspond to drier conditions. E Walsh-
Lawler Seasonality Index32 and F normalized inter-annual precipitation variability
(Methods) based on ERA5 data (monthly, 1981–202133). See Supplementary Fig. S1
for a similar map of MODIS NDVI. from WorldCLIM31. Note that higher values correspond to drier conditions. E Walsh-
Lawler Seasonality Index32 and F normalized inter-annual precipitation variability
(Methods) based on ERA5 data (monthly, 1981–202133). See Supplementary Fig. S1
for a similar map of MODIS NDVI. Fig. 1 | Spatial distribution of data used in this study. Long-term median
A vegetation optical depth (VOD, 1987–2017,23) and B normalized difference
vegetation index (GIMMS3g NDVI, 1981–2015,24). C IGBP Land-cover classes46,
masked for anthropogenic influence (Methods). D Global aridity index, adapted metrics are important because they yield spatially homogeneous
resilience metrics that also cover locations where no empirical recov-
ery rate can be fitted reliably. Moreover, although not done here,
variance-and AC1-based estimates in principle allow changes in resi-
lience over time to be quantified. and MODIS MOD1325 (see Methods for details on each dataset). Note
that the first two vegetation datasets rely upon merging data from
different satellite sensors, whereas the latter stems from continuous
measurements by a single sensor; potential impacts of the merging of
data from different sensors on our results can thereby be controlled. Combining data from different satellite sensors can lead to biases in
estimates of the temporal changes of resilience indicators due to
induced time-varying changes in the higher-order statistics of the
resulting times series26. Here, however, we only compute resilience
indicators over the full available time spans of each dataset; in this
case, a changing satellite composition does not induce systematic
biases in our resilience estimates (Methods). Verbesselt et al.1 showed that in the tropics, the AC1 of vegetation
systems has an inverse relationship with mean annual precipitation
(MAP), suggesting that vegetation in wetter regions is more resilient. Here, we rely on both empirical and theoretical resilience metrics to
investigate the effects of water availability and variability—quantified
over multiple time scales from seasonal, annual, to multi-annual—on
vegetation resilience globally, i.e., for all climate zones and land-
cover types. We analyze the dependence of resilience on aridity (Fig. 1D,
Methods)—an estimate of water surplus or deficit—and both intra- and
inter-annual precipitation variability (Fig. 1E, F, Methods) and also
investigate differences in these relationships for varying land-cover
types. Global vegetation resilience linked to water
availability and variability Under some assumptions,
it can indeed be shown analytically that the variance and AC1 are
related to the recovery rate and hence the resilience of the system in
question; an empirical confirmation of these relationships—and
thereby an empirical justification for the use of variance and AC1 as
proxies for vegetation resilience—has recently been provided using
global-scale satellite data4. Based on the results from the latter study,
we will in the following focus on three different ways of estimating
vegetation resilience. The direct empirical recovery rate obtained from
fitting an exponential recovery model to vegetation time series after
experiencing abrupt perturbations will be compared to the theoretical
recovery rate estimates inferred from both variance and AC1 (see
Methods). The empirical recovery rate is important because it gives a
directly measurable resilience metric for those locations where per-
turbations occurred; on the other hand, the variance- and AC1-based Nature Communications| (2023) 14:498 Nature Communications| (2023) 14:498 Nature Communications| (2023) 14:498 1 https://doi.org/10.1038/s41467-023-36207-7 Article 0.5
1.0
1.5
Long-term VOD Median (1987-2017)
(A)
(B)
(C)
(D)
0.2
0.4
0.6
0.8
Long-term NDVI Median (1981-2015)
0
1
2
3
WorldCLIM Aridity Index (PET/P)
0.2
0.4
0.6
0.8
1.0
1.2
Walsh-Lawler Seasonality Index (1981-2020)
(E)
0.0
0.2
0.4
0.6
0.8
Normalized Inter-Annual Precipitation Variability (1981-2020)
(F)
Evergreen (Needle)
Evergreen (Broadleaf)
Deciduous
(Needle)
Deciduous
(Broadleaf)
Open
Shrublands
Closed
Shrublands
Woody Savanna
Savanna
Grasslands
Mixed Forests
IGBP Land Cover Classification
Fig. 1 | Spatial distribution of data used in this study. Long-term median
A vegetation optical depth (VOD, 1987–2017,23) and B normalized difference
vegetation index (GIMMS3g NDVI, 1981–2015,24). C IGBP Land-cover classes46,
masked for anthropogenic influence (Methods). D Global aridity index, adapted
from WorldCLIM31. Note that higher values correspond to drier conditions. E Walsh-
Lawler Seasonality Index32 and F normalized inter-annual precipitation variability
(Methods) based on ERA5 data (monthly, 1981–202133). See Supplementary Fig. S1
for a similar map of MODIS NDVI. Article
https://doi.org/10.1038/s41467-023-36207-7 (B)
0.2
0.4
0.6
0.8
Long-term NDVI Median (1981-2015) 0.5
1.0
1.5
Long-term VOD Median (1987-2017)
(A) (B) (A) (D)
0
1
2
3
WorldCLIM Aridity Index (PET/P) (C)
Evergreen (Needle)
Evergreen (Broadleaf)
Deciduous
(Needle)
Deciduous
(Broadleaf)
Open
Shrublands
Closed
Shrublands
Woody Savanna
Savanna
Grasslands
Mixed Forests
IGBP Land Cover Classification (C) (D) 1
2
WorldCLIM Aridity Index (PET/P) 0.0
0.2
0.4
0.6
0.8
Normalized Inter-Annual Precipitation Variability (1981-2020)
(F) 0.2
0.4
0.6
0.8
1.0
1.2
Walsh-Lawler Seasonality Index (1981-2020)
(E) (F) (E) from WorldCLIM31. Nature Communications| (2023) 14:498 Global vegetation resilience linked to water
availability and variability Importantly, we consider the relationships between the To investigate global vegetation resilience patterns, we use and
compare three different vegetation datasets: long-term satellite-
derived vegetation optical depth (VOD) (Fig. 1A)23, as well as normal-
ized difference vegetation index (NDVI) data from GIMMS3g (Fig. 1B24) Nature Communications| (2023) 14:498 2 Article https://doi.org/10.1038/s41467-023-36207-7 Fig. 2 | Comparison of aridity and intra- and inter-annual precipitation varia-
bility in their relative importance for vegetation resilience at the global scale. A, B Vegetation optical depth (VOD), C, D GIMMS3g normalized difference vege-
tation index (NDVI), and E, F MODIS NDVI. Aridity compared to intra-annual (left
column) and inter-annual (right column) precipitation variability. Hexbins colored
by recovery rate computed from AC1 (minimum five points per bin). Values of the
recovery rate λ closer to zero imply lower resilience. Transition from water surplus
(aridity <1) to deficit marked with dashed vertical line; there is a sharp increase in
resilience as water availability increases. Higher inter-annual precipitation varia-
bility (right column) consistently leads to lower resilience; intra-annual precipita-
tion variability, i.e., seasonality, has a more varied impact. See Supplementary
Fig. S3 for a direct comparison of intra- and inter-annual precipitation variability. Fig. 2 | Comparison of aridity and intra- and inter-annual precipitation varia-
bility in their relative importance for vegetation resilience at the global scale. A, B Vegetation optical depth (VOD), C, D GIMMS3g normalized difference vege-
tation index (NDVI), and E, F MODIS NDVI. Aridity compared to intra-annual (left
column) and inter-annual (right column) precipitation variability. Hexbins colored
by recovery rate computed from AC1 (minimum five points per bin). Values of the recovery rate λ closer to zero imply lower resilience. Transition from water surplus
(aridity <1) to deficit marked with dashed vertical line; there is a sharp increase in
resilience as water availability increases. Higher inter-annual precipitation varia-
bility (right column) consistently leads to lower resilience; intra-annual precipita-
tion variability, i.e., seasonality, has a more varied impact. See Supplementary
Fig. S3 for a direct comparison of intra- and inter-annual precipitation variability. between months) to high (annual precipitation is concentrated in a
short period) (Fig. 1E); and (3) the year-to-year variability of rainfall,
which we define as the normalized standard deviation of annual pre-
cipitation (AP) sums. As total MAP and the standard deviation of AP are
—as should be expected—highly correlated (Supplementary Fig. Results To measure resilience, we rely here on a direct empirical quanti-
fication of resilience in terms of the recovery rate from large
perturbations4, as well as two different theory-based estimates of the
restoring rate λ, derived from the AC1 and from the variance4. We note
that we define the recovery rate λ as a negative number; values closer
to zero imply slower recovery and hence lower resilience; corre-
spondingly, higher AC1 and variance values imply lower resilience (see
Methods). Vegetation structure and productivity are tightly coupled to both
short- and long-term water availability27. Differences in annual pre-
cipitation sums have been proposed as a control on vegetation resi-
lience; e.g., it has been shown that the AC1 computed from the NDVI in
tropical rainforests is negatively correlated with MAP1, suggesting
lower resilience in drier places. However, the relationship between
AC1-based resilience estimates and MAP has not been documented
globally. Furthermore, vegetation growth and health does not only rely
on the amount of water available, but also on the consistency of that
water availability28–30—even intermittent periods of water deficit will
negatively impact plant functioning and growth. Global vegetation resilience linked to water
availability and variability S2),
we normalize the standard deviation of AP by the MAP pixelwise,
giving a suitable normalized inter-annual precipitation variability
estimate (Fig. 1F). Further, we consider a reanalysis-based soil moisture
estimate as an additional proxy for plant-available water33. different climatic predictors and vegetation resilience separately for
each land-cover type; this assures that differences in resilience indi-
cators caused by different land-cover types are not mistaken for dif-
ferences in their actual resilience. Our analysis extends the discussion
of vegetation resilience and its dependence on long-term precipitation
characteristics to the global scale and uncovers succinct and variable
relationships between water availability and both theoretical and
recently introduced empirical4 measures of vegetation resilience. Nature Communications| (2023) 14:498 Long-term water availability The revealed concise relationship between aridity and resilience
(Fig. 2) is broadly consistent across most land-cover types (Fig. 3). Resilience tends to increase non-linearly with increasing water avail-
ability; some land covers show distinctly stronger aridity/AC1-derived λ
relationships as landscapes transition from water-balance (aridity ~ 1)
to water deficit (aridity >1). For example, Savannas (olive line, Fig. 3)
show a sharp transition at aridity ~ 1 for MODIS NDVI (the linear slope
for aridity <1 vs empirical λ (AC1 λ) is 0.26 (0.18), compared to 0.13
(0.05) for aridity >1). For both VOD and NDVI, vegetation resilience
changes in many land-cover types plateau above aridity >2, which
roughly demarcates the transition into semi-arid environments31. In
these regions, grass and savanna landscapes dominate (Fig. 3); it is
likely that plant adaptations to water limitations34 account for some of
this asymptotic behavior. y
y
p
It should be noted that we do not see a decrease in vegetation
resilience when considering the inter-annual precipitation variability
without normalizing by the total annual precipitation sums (Supple-
mentary Fig. S10); higher absolute precipitation variability estimates
correspond to higher vegetation resilience. This positive correlation is
driven primarily by the absolute MAP itself; the standard deviation of
annual precipitation sums is higher in regions that have overall higher
MAP, since it cannot be negative. If we limit our analysis to only a small
precipitation range around the median (40–60th percentile of annual
precipitation values by land cover) to account for this in an alternative
way to normalizing, the pattern is the same as for the normalized inter-
annual precipitation variability. Namely, we find lower resilience for
higher precipitation variability (Supplementary Fig. S11), even if we do
not normalize inter-annual precipitation variability by MAP (Supple-
mentary Fig. S12). The relationships between aridity and resilience are overall con-
sistent for the NDVI and VOD data, but not identical. While it is not
possible to identify a single cause across all ecosystems, differences in
the attributes measured by NDVI and VOD, respectively—and the
intrinsic internal variability of each dataset—likely drive the hetero-
geneity. It should be noted that NDVI reflects vegetation chlorophyll
content or photosynthetic activity, whereas VOD reflects vegetation
density and productivity. Article ecosystems have adapted to annual and short-term water deficits with
a variety of methods (for example, phenotypic plasticity, drought
pruning)34,36,37. In contrast, inter-annual and longer-term water deficits
can cause large shifts in vegetation (and ecosystem) species mixes36. It
is also important to note that inter-annual precipitation variability is
not static; recent research has found spatially heterogeneous changes
in water deficits (i.e., drought events)9,35,38. Our results indicate that
increasingly frequent and extreme water deficits—especially those at
the multi-annual scale—will impact the resilience of vegetation eco-
systems worldwide. Such events will have a relatively larger impact
than intra-annual precipitation variability on vegetation resilience as
ecosystems lose the ability to recover to their previous state. highly seasonal precipitation. In contrast, high inter-annual precipita-
tion variability (Fig. 2, right column) leads to almost universally lower
resilience, indicating that more consistent precipitation year-on-year
encourages more resilient vegetation. At the global scale, we thus infer
a clear increase in vegetation resilience with increasing water avail-
ability and with decreasing inter-annual precipitation variability
(Fig. 2B, D, F); the relationship between resilience and precipitation
seasonality is less concise (Fig. 2A, C, E). Long-term water availability While some differences regarding the esti-
mated resilience should therefore be expected, the overall similarity
between the results obtained for the three data sources provides a
strong argument that the inferred reduction of resilience with higher
aridity across land-cover types is robust. In particular, the fact that we
obtain similar results for the single-sensor MODIS NDVI as for the other
two, mixed-sensor datasets, implies that the merging of signals for the
latter data products do not affect our results. Discussion Water availability plays a primary role in controlling the occurrence,
type, and health of vegetation globally. Our work extends that of
previous research1, and documents a global relationship of less resi-
lient vegetation with lower water availability (Fig. 1), lower MAP (Sup-
plementary Fig. S4), and lower soil moisture (Supplementary Fig. S5). This relationship is consistent across land-cover types, with many land
covers showing the greatest changes in vegetation resilience as aridity
approaches 1 (i.e., a balance between precipitation and potential eva-
potranspiration) (Figs. 2, 3). The total yearly amount of precipitation, however, is not the only
control on the health and resilience of vegetation. We find a globally
consistent
pattern—particularly
in
grass-dominated
regions—of
decreasing vegetation resilience with higher inter-annual precipitation
variability (Fig. 5). While more work is required to fully constrain the
mechanisms behind this response, we posit that vegetation reliant on
surface water and direct precipitation is more strongly impacted
during low-precipitation periods than tree-dominated areas with
deeper root systems and access to longer-term water storage (e.g.,
lakes, rivers, and shallow groundwater). It has previously been shown
that vegetation productivity is tightly coupled to antecedent pre-
cipitation in arid to semi-arid environments30 and that regions with
higher woody biomass are relatively buffered against short-term water
deficits29,35. Our results confirm these observations at the global scale,
and establish an additional link between precipitation variability and
vegetation resilience. Nature Communications| (2023) 14:498 Joint effects of water availability and precipitation variability on
vegetation resilience globally To assess the first-order relationships between vegetation resilience
and water availability, we first consider all land-cover types and climate
zones together (Fig. 2). For all three vegetation indices (VOD and two
NDVI datasets), we find that the highest recovery rates are generally
found in areas of water surplus (Fig. 2, left of dashed line). We consider three ways of globally measuring water availability
and variability as drivers of vegetation resilience, encompassing mul-
tiple overlapping time scales: (1) the Aridity Index (Fig. 1D)31, which
provides a measure of long-term MAP relative to potential evapo-
transpiration; we consider it more appropriate to consider aridity
rather than, e.g., MAP, in order to make the results comparable across
different climate and vegetation zones. In addition, we consider two
measures of intra- and inter-annual rainfall variability: (2) the Walsh-
Lawler seasonality index32, which measures how precipitation is dis-
tributed throughout the year—from low (precipitation is similar The relationship to shorter-term precipitation variability is less
clear, however. Intra-annual precipitation variability does not scale
cleanly with recovery rate (Fig. 2, left column); there exist highly resi-
lient areas which receive precipitation only during short time periods. These areas are found exclusively in grass and shrublands globally and
are concentrated in the African Sahel, where plants are adapted to Nature Communications| (2023) 14:498 3 https://doi.org/10.1038/s41467-023-36207-7 Article Intra- and inter-annual precipitation variability While there is a clear demarcation between the resilience of vegetation
in water-surplus and water-deficit regions at the global scale (Fig. 2)
and when separated by land cover (Fig. 3), the role of intra-annual
water variability is not as clear (Fig. 4). We find that resilience is broadly
similar across precipitation seasonalities; however, Kendall-Tau coef-
ficients remain generally positive, implying decreasing resilience with
more seasonal precipitation. Relationships vary across land-cover
types; grass-dominated landscapes in particular (woody savanna,
savanna, grasslands) have regions of both positive and negative rela-
tionships between resilience and seasonality. We posit this is due to the
wide distribution of these land-cover zones across the globe (Fig. 1C)—
woody savannas are dominant in both central Africa and at high
northern latitudes in Canada and Siberia. Vegetation is not only sensitive to the distribution of precipitation
within the year, but also to its distribution between years30,35. Across all
three datasets and measures of resilience, we find that higher relative
inter-annual variability of precipitation leads to less resilient vegeta-
tion, particularly in grass-dominated landscapes (Fig. 5, Supplemen-
tary Fig. S8). We further find that inter-annual precipitation variability
(Fig. 5) is a relatively stronger control on resilience than intra-annual
precipitation distribution (Fig. 4). We posit that this difference is due
to the characteristic time scales at which vegetation responds to water
deficits, with longer (inter-annual) time scales being relatively harder
to adapt to. In regions with highly seasonal precipitation, vegetated An important caveat is that we do not examine long-term changes
in precipitation or climatic conditions. All else being equal, changes in
precipitation will engender a change in vegetation, which generally will
be expressed in our resilience metrics. We aim in this study to map
long-term global patterns; our analysis framework here does not allow
us to disentangle whether vegetation resilience has changed4, and to
what degree those changes are driven by precipitation changes. We further note that differences in resilience between land-cover
types likely reflect mainly intrinsic, physiological differences in vege-
tation—with all other drivers fixed, vegetation will grow more quickly
in a rainforest than in a savanna. The measured differences also Nature Communications| (2023) 14:498 4 Article https://doi.org/10.1038/s41467-023-36207-7 how well satellite data products capture fine-scale changes
on of different densities and structures. Our discussion
ses on the relationship between vegetation resilience and
bility within single land-cover classes. Nature Communications| (2023) 14:498 Vegetation and land-cover data To monitor vegetation at the global scale, we use three datasets: (1)
vegetation optical depth (VOD, 0.25°, Ku-Band, daily 1987–201723)
(Fig. 1A), (2) AVHRR GIMMSv3g normalized difference vegetation
index (NDVI, 1/12°, bi-weekly 1981–201524) (Fig. 1B), and (3) MODIS
MOD13 NDVI at 0.05° (16-day, 2000–202125). We correct for spurious
values in the NDVI data (e.g., cloud contamination) using the method
of Chen et al.43. We resample the VOD data using bi-weekly medians to
agree with the NDVI data time sampling. For all three vegetation datasets, we remove seasonality and long-
term trends using seasonal trend decomposition by Loess4,44 based on
the proposed optimal parameters listed in Cleveland et al.44 (code
available on Zenodo45). That is, we use a period of 24 (bi-monthly, 1
year), 47 for the trend smoother (just under 2 years) and 25 for low-
pass (just over 1 year). We only use the STL residual—the de-seasoned
and de-trended NDVI and VOD time series—in our analysis. Our analysis rests on both empirical recovery rates from per-
turbations – direct estimates of vegetation resilience according to
the common definition—and two theory-based estimates (from AC1
and variance) that are hence more indirect. Although our results are
globally coherent, we therefore note that especially our two theory-
based resilience estimates may, in principle, be influenced by local-
scale factors. For example, the varying physiological characteristics
of different vegetation types contained within each time series will
present a mixed signal in many regions. We have minimized this
influence by performing our analyses separately for different land
covers, and by limiting our work to natural land covers. However, it is
possible that some regions retain mixed signals (cf. Fig. 4), which
could influence local-scale resilience estimates. At the global scale,
however, we find overall consistent behavior across climate and
vegetation zones. Using VOD at the global scale, we have previously
empirically confirmed the theoretical relationships between AC1 and
variance on the one hand, and empirically inferred recovery rates on
the other hand, suggesting that the AC1 and variance can indeed
serve as resilience metrics4. The consistency between our results for
the empirical recovery rates and the theory-based estimates across
land-cover types adds strong further independent evidence to this. Precipitation data and variability metrics To measure precipitation at the global scale, we rely upon ERA5 data
(~30 km, monthly, 1981–2021)33. We process global-scale precipitation
metrics using the Google Earth Engine47 platform. We further use the
sum of soil moisture from the surface down to 28 cm of depth (first
two layers of the ECMWF Integrated Forecasting System soil moisture
estimates)
to
quantify
soil
moisture
means
and
inter-annual
variability33. It is well-documented that vegetation resilience is responsive to
the MAP of certain regions1. However, the role of precipitation varia-
bility in controlling vegetation resilience has not been well-studied. Here we examine precipitation variability in terms of both intra- and
inter-annual patterns. Intra-annual precipitation variability is deter-
mined in terms of the Walsh-Lawler Seasonality index32 (Fig. 1D), cal-
culated using monthly data from ERA533. g
y
Partly due to the fact that precipitation is non-negative, simple
inter-annual variability metrics such as the standard deviation of
annual precipitation sums are biased by the absolute precipitation
sums; higher precipitation regions have a higher possible range of
variability. To limit the influence of MAP, we hence investigate the
standard deviation of annual precipitation sums normalized by the
MAP, over the period 1981–2021, based on ERA5 data33 (Fig. 1F). We
motivate our normalization by MAP with the strong linear rela-
tionship between MAP and MAP standard deviation (Supplementary
Fig. S2). We further confirm our discovered relationships (Fig. 5)
using only those regions where MAP was between the 40 and 60th
percentile
of
MAP
for
a
given
land
cover
(Supplementary
Figs. S11,S12). This serves as an additional check that our normal-
ization of MAP standard deviation by MAP does not bias the inferred
relationship between vegetation resilience and precipitation varia-
bility. Similarly, we generate a normalized inter-annual soil moisture We have presented evidence based on both empirically esti-
mated recovery rates and different theoretical—yet empirically con-
firmed—resilience metrics for concise global relationships between
vegetation resilience and water availability, modulated by land-cover
type. We find overall greater resilience in regions with higher water
availability across climate zones and vegetation types, based on an
aridity index. However, our results also suggest that resilience con-
sistently declines with increasing precipitation variability especially
on inter-annual time scales, and particularly in grass-dominated
landscapes. Simulations from the sixth phase of the Climate Model
Intercomparison Project suggest increased precipitation variability
under global warming scenarios in the coming decades. Intra- and inter-annual precipitation variability Equivalent figure for mean annual precipitation (MAP) shown as Supplementary
Fig. S4, and for mean annual soil moisture shown as Supplementary Fig. S5. Figure
for aridity showing all three instruments and metrics as Supplementary Fig. S6. insignificant relationships (p > 0.05) shown as a black circle. Additional box-plot of
1000 randomly sampled surrogates (box edges: 25–75th percentiles, black line:
median) shown with red for MODIS NDVI, orange for AVHRR NDVI24, and blue for
VOD. KT of medians consistently higher than box plots due to random sampling
(see Methods). Both VOD and NDVI exhibit lower resilience—i.e., λ closer to zero,
see Methods—with lower water availability across the majority of land-cover types. Equivalent figure for mean annual precipitation (MAP) shown as Supplementary
Fig. S4, and for mean annual soil moisture shown as Supplementary Fig. S5. Figure
for aridity showing all three instruments and metrics as Supplementary Fig. S6. Vegetation and land-cover data In
particular, the fact that we find very similar results for the empirical
recovery rates and the theory-based estimates for the MODIS NDVI
data show that this confirmation is not impacted by merging data
from different sensors. To contextualize our understanding of vegetation resilience, we
use MODIS MCD12Q1 land cover46 (Fig. 1C) as well as a global average
aridity index based on WorldCLIM data31 (Fig. 1D).We exclude from our
analysis anthropogenic and non-vegetated landscapes (e.g., perma-
nent snow and ice, desert, urban), as well as any land covers which have
changed (e.g., forest to grassland) during the period 2001–2020. Intra- and inter-annual precipitation variability Despite this control,
ere remain large differences in vegetation within the same
l
t th
l b l
l
O
lt (Fi
2 5) th
precipitation variability; further and smaller-scale wo
required to constrain how different plant responses to
variability
impact
their
resilience
to
changing
e
conditions. Despite considerable spatial heterogeneity39, a large
i t t
i
i
i it ti
i bilit i
th
https://doi.org/10.1038/s41467 depend on how well satellite data products capture fine-scale changes
in vegetation of different densities and structures. Our discussion
hence focuses on the relationship between vegetation resilience and
water availability within single land-cover classes. Despite this control,
however, there remain large differences in vegetation within the same
broad land-cover classes at the global scale. Our results (Figs. 2–5) thus
include an array of different vegetation mixes and responses to precipitation variability; further and smaller-scale work would be
required to constrain how different plant responses to precipitation
variability
impact
their
resilience
to
changing
environmental
conditions. Despite considerable spatial heterogeneity39, a large body of work
points to increasing precipitation variability in the coming decades in
response to anthropogenic climate change9. The global tendency is for Nature Communications| (2023) 14:498 5 https://doi.org/10.1038/s41467-023-36207-7 Article Fig. 3 | Vegetation resilience as a function of aridity31 at the global scale,
separated by land-cover type46. Vegetation resilience λ estimated empirically
(A, B) and via the AC1 (C, D) for vegetation optical depth (VOD, left column) and
MODIS NDVI (right column). Binned medians shown as solid dots (Kendall-Tau (KT)
p <0.05) and transparent arrows (KT p>0.05), with 25–75th percentiles of each bin
shown as connected vertical lines capped with hatches. Land covers with less than
1000 points or less than 10 bins of at least 50 members are omitted. E KT coeffi-
cients (aridity vs AC1-derived λ, panels C, D) for each land-cover type. Significant
(p < 0.05) KTs shown as a black triangle (KT of median binned data, cf. C, D), insignificant relationships (p > 0.05) shown as a black circle. Additional box-plot of
1000 randomly sampled surrogates (box edges: 25–75th percentiles, black line:
median) shown with red for MODIS NDVI, orange for AVHRR NDVI24, and blue for
VOD. KT of medians consistently higher than box plots due to random sampling
(see Methods). Both VOD and NDVI exhibit lower resilience—i.e., λ closer to zero,
see Methods—with lower water availability across the majority of land-cover types. Methods
Vegetation and land-cover data wetter regions to receive more water and drier regions to become
drier, as well as for increases in both wet and dry extremes9,35,40. Based
on our research, it is clear that changes in both precipitation volume
and variability will have a measurable and spatially heterogeneous
impact
on
global
vegetation
(Figs. 2–5). Low-
and
variable-
precipitation
regions
will
face
comparably
higher
burdens
in
response to increasing precipitation variability in a warming world; this
shift has strong implications for future ecosystem functioning in vast
parts of the sub- and extra-tropics. Resilience loss, and eventually
potential desertification of grass- and shrublands, could trigger a chain
of destabilizing feedbacks; for example, vegetation resilience loss and
ecosystem transitions could reduce water storage capabilities at con-
tinental scales41, affect rainfall patterns due to atmosphere-vegetation
interactions8, and accelerate greenhouse gas emissions11,42. wetter regions to receive more water and drier regions to become
drier, as well as for increases in both wet and dry extremes9,35,40. Based
on our research, it is clear that changes in both precipitation volume
and variability will have a measurable and spatially heterogeneous
impact
on
global
vegetation
(Figs. 2–5). Low-
and
variable-
precipitation
regions
will
face
comparably
higher
burdens
in
response to increasing precipitation variability in a warming world; this
shift has strong implications for future ecosystem functioning in vast
parts of the sub- and extra-tropics. Resilience loss, and eventually
potential desertification of grass- and shrublands, could trigger a chain
of destabilizing feedbacks; for example, vegetation resilience loss and
ecosystem transitions could reduce water storage capabilities at con-
tinental scales41, affect rainfall patterns due to atmosphere-vegetation
interactions8, and accelerate greenhouse gas emissions11,42. Precipitation data and variability metrics Based on our
empirical results we hence infer an increasing risk of vegetation
degradation and eventually desertification—especially in regions with
savanna, grass- and shrublands—in response to anthropogenic cli-
mate change. Nature Communications| (2023) 14:498 6 Article https://doi.org/10.1038/s41467-023-36207-7 by normalizing year-on-year soil moisture standard
upplementary Fig. S8) by long-term mean soil moisture
tary Fig. S5). esilience estimation
defined as the ability of a system to recover from per-
to the previous state16,17. To measure resilience on the g
employ a recently introduced methodology4 which
summarize in the following. We first identify sharp transitions in the vegetat
using an 18-point (9 month) moving window to defi
throughout the time series48. We then identify slopes
https://doi.org/10.1038/s4146 variability by normalizing year-on-year soil moisture standard
deviation (Supplementary Fig. S8) by long-term mean soil moisture
(Supplementary Fig. S5). variability by normalizing year-on-year soil moisture standard
deviation (Supplementary Fig. S8) by long-term mean soil moisture
(Supplementary Fig. S5). to the previous state16,17. To measure resilience on the global scale, we
employ a recently introduced methodology4 which we will briefly
summarize in the following. We first identify sharp transitions in the vegetation time series
using an 18-point (9 month) moving window to define local slopes
throughout the time series48. We then identify slopes above the 99th
percentile, and define connected regions as individual perturbations. Empirical resilience estimation Empirical resilience estimation Furthermore, the
same transitions are not guaranteed to be captured for both NDVI and
VOD data in each location, as the percentiles will naturally vary
between the datasets. Finally, our method will in some cases produce
false positives, especially in cases where a given time series does not
have any significant rapid transitions. To limit the influence of false
positives on our results, we discard any perturbations where the
time series does not drop significantly, and where the period
before and after a given transition does not pass a two-sample
Kolmogorov–Smirnov test4. hx2i = σ2
2λ
ð3Þ
for the variance, and
αðnÞ = enλΔt
ð4Þ hx2i = σ2
2λ
ð3Þ
for the variance, and
αðnÞ = enλΔt
ð4Þ hx2i = σ2
2λ
ð3Þ
for the variance, and ð3Þ αðnÞ = enλΔt
ð4Þ ð4Þ for the AC1. Hence, the closer λ is to zero, the larger the AC1 and
variance, corresponding to lower stability. Note that theory55 suggests
that the recovery rate r is equal to the restoring rate λ. An empirical
global confirmation for this relationship for has recently been
demonstrated based on both NDVI and VOD data4. We compare the
dependence of the recovery rate r and two different theoretical esti-
mates of the restoring rate λ—obtained via inverting the above equa-
tions for the variance and AC1—with respect to their dependence on
water availability and its variability. Finally, using our global set of time-series transitions, we can
identify each local vegetation (NDVI or VOD) minima, and use the five
following years of data to fit an exponential function to the residual
time series, assuming that the recovery after a perturbation to a
vegetation state x0 follows approximately the equation It is important to note that combining data from different sensors
with varying signal-to-noise ratios (e.g., VOD, AVHRR NDVI) can bias
estimates of temporal changes in resilience indicators because the
higher-order
statistics
of
the
resulting
time
series
are
not
homogeneous4,26. In the present work, however, we do not investigate
temporal trends via estimating resilience indicators in sliding windows
(as in refs. 4,26), but rather estimate resilience indicators for the full
available time series. This excludes the possibility of systematic biases
in our AC1- and variance-based estimates of the restoring rate λ. Empirical resilience estimation Resilience is defined as the ability of a system to recover from per-
turbations, and can be quantified empirically by the speed of recovery 7 Nature Communications| (2023) 14:498 7 Article https://doi.org/10.1038/s41467-023-36207-7 Fig. 4 | Vegetation resilience as a function of precipitation seasonality in terms
of the Walsh-Lawler seasonality index32 (Methods), separated by land-cover
type46. Vegetation resilience λ estimated empirically (A, B) and via the AC1 (C, D)
for vegetation optical depth (VOD, left column) and MODIS NDVI (right column). Binned medians shown as solid dots (Kendall-Tau (KT) p < 0.05) and transparent
arrows (KT p>0.05), with 25–75th percentiles of each bin shown as connected
vertical lines capped with hatches. Land covers with less than 1000 points or less
than 10 bins of at least 50 members are omitted. E KT coefficients (aridity vs AC1-
derived λ, panels C, D) for each land-cover type. Significant (p < 0.05) KTs shown as
a black triangle (KT of median binned data, cf. C, D), insignificant relationships (p > 0.05) shown as a black circle. Additional box-plot of 1000 randomly sampled
surrogates (box edges: 25–75th percentiles, black line: median) shown with red for
MODIS NDVI, orange for AVHRR NDVI24, and blue for VOD. KT of medians con-
sistently higher than box plots due to random sampling (see Methods). Both VOD
and NDVI exhibit lower resilience—i.e. λ closer to zero, see Methods—with lower
water availability across the majority of land-cover types. While for all three con-
sidered vegetation datasets empirical recovery rates generally decrease with more
concentrated precipitation, the relationship between Walsh-Lawler seasonality and
recovery rates is less steep than for aridity (Fig. 3). Figure showing all three
instruments and metrics as Supplementary Fig. S7. discretizing the resulting Ornstein-Uhlenbeck process into time steps
of width Δt, the variance and AC1 of the resulting order-one
autoregressive process are then related to the restoring rate λ via54: The highest peak (largest instantaneous slope) within each connected
region is then labeled as an individual disturbance. discretizing the resulting Ornstein-Uhlenbeck process into time steps
of width Δt, the variance and AC1 of the resulting order-one
autoregressive process are then related to the restoring rate λ via54: The employed approach does not delineate every rapid transition
in a time series due to our reliance on percentiles; our dataset will be
inherently biased towards the largest transitions. Binning and significance testing The direction and magnitude of our discovered relationships (e.g.,
Figs. 3–5, Supplementary Figs. S4–S12) will be to some degree con-
trolled by the choice of bin sizes. We tested three bin sizes for each
different variable (Supplementary Figs. S13,S14). We also imposed the
conditions that there were at least 50 measurements in each bin to
form a proper median, and that we only report those relationships
which cover ten or more bins. Empirical resilience estimation In
principle, the combination of different sensors might lead to larger
uncertainties in the estimates of λ: combining different sensor data
leads to temporally varying (yet spatially homogeneous) effects on the
AC1 and variance and may therefore lead to a wider spread—but not a
bias—in the resulting estimates of λ. xðtÞ ≈x0ert
ð1Þ ð1Þ where x(t) denotes the vegetation state at time t after the perturbation. Negative r indicates that the vegetation system will return to the ori-
ginal stable state at rate ∣r∣. For positive r, the initial perturbation would
be amplified, suggesting a non-resilient vegetation state. Our empirical
recovery rates are defined as the fitted exponent r, obtained for each
detected transition in the NDVI and VOD residual time series. We finally
use the coefficient of determination R2 to remove instances where the
fitted exponential poorly matches the underlying data4. For the empirical estimate of the restoring rate obtained from
fitting an exponential to the recovery after an abrupt negative devia-
tion of VOD or NDVI, abrupt changes in the mean state induced by
changing sensors rather than an actual vegetation shift may impact the
results. However, all datasets used here are tightly cross-calibrated to
eliminate mean-shifts when new instruments are introduced23,24. It is
therefore unlikely that changes in the instrumentation of the various
datasets unduly influence our empirical estimates of λ. Dynamical system metrics of resilience The lag-one autocorrelation (AC1) has previously been proposed to
measure the stability of real-world dynamical systems in general, and
the resilience of vegetation systems in particular1,19–21,49. Based on the
concept of critical slowing down, the AC1 has, together with the var-
iance, also been suggested as an early-warning indicator for forth-
coming critical transitions50,51. Mathematically, the suitability of the
variance and AC1 as resilience measures and early-warning indicators
can be motivated as follows4,52,53. First, linearize the system around a
given stable state x*: To better constrain the relationship between each driving vari-
able and resilience, we use the non-parametric Kendall-Tau test56. Kendall-Tau statistics are calculated over each set of binned medians,
as well as using a Monte-Carlo approach. Over 1000 iterations, we
choose one random point from each bin and recalculate the Kendall-
Tau statistics. These 1000 surrogates are displayed as box plots in
Figs. 3–5. Note that the Kendall-Tau value of the median line will
almost always be larger than the median of the 1000 Kendall-Tau
values resulting from the surrogates due to the smoothing inherent in
taking binned medians. That is, the binned medians represent a
smooth and almost monotonic line with fewer jumps, while the
1000 surrogates will have strong fluctuations from one bin to the dx = λxdt + σdW
ð2Þ dx = λxdt + σdW
ð2Þ ð2Þ dx = λxdt + σdW for x := x x*, assuming a Wiener Process W with standard deviation σ. The dynamics are stable for λ < 0 and unstable otherwise. Upon for x := x x*, assuming a Wiener Process W with standard deviation σ. The dynamics are stable for λ < 0 and unstable otherwise. Upon Nature Communications| (2023) 14:498 8 Nature Communications| (2023) 14:498 Article https://doi.org/10.1038/s41467-023-36207-7 ng to overall lower Kendall-Tau values. The fraction of
u statistics which share a sign with the Kendall-Tau of the
is also reported on Supplementary Figs. S4–S14. d that our Kendall-Tau statistics are robust against changing
upplementary Figs. S13,S14), where the direction of trends
hange from those reported in Fig. 3. The magnitude of the
u statistic as well as p values shift with different bin sizes
with smaller bin sizes typically resulting in more rob
Changes in bin size do not have a strong impact upon ou
pretations or conclusions. Dynamical system metrics of resilience Reporting summary
Further information on research design is available in
Portfolio Reporting Summary linked to this article
https://doi.org/10.1038/s41467-0 next, leading to overall lower Kendall-Tau values. The fraction of
Kendall-Tau statistics which share a sign with the Kendall-Tau of the
median line is also reported on Supplementary Figs. S4–S14. next, leading to overall lower Kendall-Tau values. The fraction of
Kendall-Tau statistics which share a sign with the Kendall-Tau of the
median line is also reported on Supplementary Figs. S4–S14. with smaller bin sizes typically resulting in more robust trends. Changes in bin size do not have a strong impact upon our data inter-
pretations or conclusions. We find that our Kendall-Tau statistics are robust against changing
bin sizes (Supplementary Figs. S13,S14), where the direction of trends
does not change from those reported in Fig. 3. The magnitude of the
Kendall-Tau statistic—as well as p-values—shift with different bin sizes, Nature Communications| (2023) 14:498 References 21. Liu, Y., Kumar, M., Katul, G. G. & Porporato, A. Reduced resilience as
an early warning signal of forest mortality. Nat. Clim. Change 9,
880–885 (2019). 1. Verbesselt, J. et al. Remotely sensed resilience of tropical forests. Nat. Clim. Change 6, 1028–1031 (2016). 1. Verbesselt, J. et al. Remotely sensed resilience of tropical forests. Nat. Clim. Change 6, 1028–1031 (2016). 2. Lovejoy, T. E. & Nobre, C. Amazon tipping point. Sci. Adv. 4,
1–2 (2018). 2. Lovejoy, T. E. & Nobre, C. Amazon tipping point. Sci. Adv. 4,
1–2 (2018). 22. Van Nes, E. H. & Scheffer, M. Slow recovery from perturbations as a
generic indicator of a nearby catastrophic shift. Am. Nat. 169,
738–747 (2007). 3. Hubau, W. et al. Asynchronous carbon sink saturation in African and
Amazonian tropical forests. Nature 579, 80–87 (2020). 23. Moesinger, L. et al. The global long-term microwave vegetation
optical depth climate archive (vodca). Earth Syst. Sci. Data 12,
177–196 (2020). 4. Smith, T., Traxl, D. & Boers, N. Empirical evidence for recent global
shifts in vegetation resilience. Nat. Clim. Change 12,
477–484 (2022). 24. Pinzon, J. E. & Tucker, C. J. A non-stationary 1981–2012 avhrr ndvi3g
time series. Remote Sens. 6, 6929–6960 (2014). 5. Boulton, C. A., Lenton, T. M. & Boers, N. Pronounced loss of amazon
rainforest resilience since the early 2000s. Nat. Clim. Change 12,
271–278 (2022). 25. Didan, K. Mod13c1 modis/terra vegetation indices 16-day l3 global
0.05deg cmg v006. NASA EOSDIS Land Processes DAAC https://
doi.org/10.5067/MODIS/MOD13C1.006 (2015). 6. Hirota, M., Holmgren, M., Van Nes, E. H. & Scheffer, M. Global
resilience of tropical forest and Savanna to critical transitions. Sci-
ence 334, 232–5 (2011). 26. Smith, T. et al. Reliability of resilience estimation based on multi-
instrument time series. Earth Syst. Dyn. Discuss. 2022, 1–14 (2022). 7. Ciemer, C. et al. Higher resilience to climatic disturbances in tro-
pical vegetation exposed to more variable rainfall. Nat. Geosci. 12,
174–179 (2019). 27. Good, S. P., Moore, G. W. & Miralles, D. G. A mesic maximum in
biological water use demarcates biome sensitivity to aridity shifts. Nat. Ecol. Evol. 1, 1883–1888 (2017). 28. Jiao, W. et al. Observed increasing water constraint on vegetation
growth over the last three decades. Nat. Commun. 12, 1–9 (2021). 8. Boers, N., Marwan, N. & Barbosa, H. M. J. A deforestation-induced
tipping point for the South American monsoon system. Sci. Rep.49,
41489 (2017). 29. Reporting summary Reporting summary
Further information on research design is available in the Nature
Portfolio Reporting Summary linked to this article. Nature Communications| (2023) 14:498 9 9 Article https://doi.org/10.1038/s41467-023-36207-7 median binned data, cf. C, D), insignificant relationships (p>0.05) shown as a black
circle. Additional box-plot of 1000 randomly sampled surrogates (box edges:
25–75th percentiles, black line: median) shown with red for MODIS NDVI, orange
for AVHRR NDVI24, and blue for VOD. KT of medians consistently higher than box
plots due to random sampling (see Methods). For both VOD and NDVI we infer
lower resilience for higher relative inter-annual precipitation variability. Equivalent
figure for normalized inter-annual soil moisture variability shown as Supplemen-
tary Fig. S8. Figure showing all three instruments and metrics as Supplemen-
tary Fig. S9. Fig. 5 | Vegetation resilience as a function of normalized inter-annual pre-
cipitation variability (Methods), separated by land-cover type46. Vegetation
resilience λ estimated empirically (A, B) and via the AC1 (C, D) for vegetation optical
depth (VOD, left column) and MODIS NDVI (right column). Binned medians shown
as solid dots (Kendall-Tau (KT) p < 0.05) and transparent arrows (KT p > 0.05), with
25–75th percentiles of each bin shown as connected vertical lines capped with
hatches. Land covers with less than 1000 points or less than 10 bins of at least 50
members are omitted. E KT coefficients (aridity vs AC1-derived λ, panels C, D) for
each land-cover type. Significant (p < 0.05) KTs shown as a black triangle (KT of median binned data, cf. C, D), insignificant relationships (p>0.05) shown as a black
circle. Additional box-plot of 1000 randomly sampled surrogates (box edges:
25–75th percentiles, black line: median) shown with red for MODIS NDVI, orange
for AVHRR NDVI24, and blue for VOD. KT of medians consistently higher than box
plots due to random sampling (see Methods). For both VOD and NDVI we infer
lower resilience for higher relative inter-annual precipitation variability. Equivalent
figure for normalized inter-annual soil moisture variability shown as Supplemen-
tary Fig. S8. Figure showing all three instruments and metrics as Supplemen-
tary Fig. S9. Data availability 13. Abis, B. & Brovkin, V. Environmental conditions for alternative tree-
cover states in high latitudes. Biogeosciences 14, 511–527 (2017). 13. Abis, B. & Brovkin, V. Environmental conditions for alternative tree-
cover states in high latitudes. Biogeosciences 14, 511–527 (2017). The Kendall-Tau statistics generated in this study have been deposited
on Zenodo at 10.5281/zenodo.743666945. The raw environmental and
satellite data used in this study are publicly available23–25,31,46. Direct
links to the datasets can be found at: GIMMS NDVI: https://www.cen. uni-hamburg.de/en/icdc/data/land/gimms-ndvi3g.html, VOD: https://
zenodo.org/record/2575599, MODIS Land Cover: https://lpdaac.usgs. gov/products/mcd12q1v006/, MODIS NDVI: https://lpdaac.usgs.gov/
products/mod13c1v006/, WorldCLIM Aridity: https://figshare.com/
articles/dataset/Global_Aridity_Index_and_Potential_Evapotranspi-
ration_ET0_Climate_Database_v2/7504448/4, ERA5 Climate Data:
https://www.ecmwf.int/en/forecasts/datasets/reanalysis-datasets/era5. 14. Feng, Y. et al. Reduced resilience of terrestrial ecosystems locally is
not reflected on a global scale. Commun. Earth Environ. 2,
1–11 (2021). 15. Forzieri, G. et al. Emerging signals of declining forest resilience
under climate change. Nature 608, 534–539 (2022). 16. Peterson, G., Allen, C. R. & Holling, C. S. Ecological re
biodiversity, and scale. Ecosystems 1, 6–18 (1998). 17. Folke, C. et al. Regime shifts, resilience, in ecosystem management. Annu. Rev. Ecol. Evol. Syst. 35, 557–581 (2004). 18. Carpenter, S. R. & Brock, W. A. Rising variance: a leading indicator of
ecological transition. Ecol. Lett. 9, 311–8 (2006). Code availability 19. Seddon, A. W., Macias-Fauria, M., Long, P. R., Benz, D. & Willis, K. J. Sensitivity of global terrestrial ecosystems to climate variability. Nature 531, 229–232 (2016). The code used for de-seasoning the data via STL, detecting abrupt
transitions, and estimating resilience both empirically and via time-
series metrics (AC1, Variance) can be found on Zenodo: https://doi. org/10.5281/zenodo.743666945. Code is written in Python (3.9.13). 20. van der Bolt, B., van Nes, E. H., Bathiany, S., Vollebregt, M. E. &
Scheffer, M. Climate reddening increases the chance of critical
transitions. Nat. Clim. Change 8, 478–484 (2018). https://doi.org/10.1038/s41467-023-36207-7 https://doi.org/10.1038/s41467-023-36207-7 56. Kendall, M. G. Rank Correlation Methods (Griffin, 1948). 34. Volaire, F. A unified framework of plant adaptive strategies to
drought: crossing scales and disciplines. Glob. Change Biol. 24,
2929–2938 (2018). Acknowledgements 35. Huang, K. & Xia, J. High ecosystem stability of evergreen broadleaf
forests under severe droughts. Glob. Change Biol. 25,
3494–3503 (2019). The State of Brandenburg (Germany) through the Ministry of Science
and Education and the NEXUS project supported T.S. for part of this
study. T.S. also acknowledges support from the BMBF ORYCS pro-
ject and the Universität Potsdam Remote Sensing computational
cluster. N.B. acknowledges funding by the Volkswagen foundation. This is TiPES contribution #208; the TiPES (‘Tipping Points in the
Earth System’) project has received funding from the European
Union’s Horizon 2020 research and innovation program under grant
agreement No. 820970. N.B. acknowledges further funding by the
European Union’s Horizon 2020 research and innovation program
under the Marie Sklodowska-Curie grant agreement No. 956170, as
well as from the Federal Ministry of Education and Research under
grant No. 01LS2001A. Open access publication funded by the
Deutsche Forschungsgemeinschaft (DFG, German Research Foun-
dation) – Projektnummer 491466077. 36. Hereford, R., Webb, R. & Longpré, C. Precipitation history and
ecosystem response to multidecadal precipitation variability in the
mojave desert region, 1893–2001. J. Arid Environ. 67, 13–34 (2006). 37. Nicotra, A. B. et al. Plant phenotypic plasticity in a changing climate. Trends Plant Sci. 15, 684–692 (2010). 38. Pokhrel, Y. et al. Global terrestrial water storage and drought
severity under climate change. Nat. Clim. Change 11,
226–233 (2021). 39. Traxl, D., Boers, N., Rheinwalt, A. & Bookhagen, B. The role of
cyclonic activity in tropical temperature-rainfall scaling. Nat. Commun. 12, 6732 (2021). 40. Lehmann, J., Mempel, F. & Coumou, D. Increased occurrence of
record-wet and record-dry months reflect changes in mean rainfall. Geophys. Res. Lett. 45, 13–468 (2018). Author contributions 41. Miralles, D. G., Gentine, P., Seneviratne, S. I. & Teuling, A. J. Land–atmospheric feedbacks during droughts and heatwaves:
state of the science and current challenges. Ann. N. Y. Acad. Sci. 1436, 19 (2019). T.S. and N.B. conceived and designed the study. T.S. processed the data
and performed the numerical analysis. Both authors interpreted the
results and wrote the manuscript. References Xu, X., Yang, D. & Sivapalan, M. Assessing the impact of climate
variability on catchment water balance and vegetation cover. Hydrol. Earth Syst. Sci. 16, 43–58 (2012). 9. Masson-Delmotte, V. et al. Ipcc, 2021: Climate change 2021: The
physical science basis. in Contribution of Working Group I to the
Sixth Assessment Report of the Intergovernmental Panel on Climate
Change. (Cambridge University Press, 2021). 30. Liu, L., Zhang, Y., Wu, S., Li, S. & Qin, D. Water memory effects and
their impacts on global vegetation productivity and resilience. Sci. Rep. 8, 1–9 (2018). 10. Lewis, S. L., Wheeler, C. E., Mitchard, E. T. & Koch, A. Restoring
natural forests is the best way to remove atmospheric carbon. Nature 568, 25–28 (2019). 31. Trabucco, A. & Zomer, R. Global Aridity Index and Potential
Evapotranspiration (ET0) Climate Database v2. figshare. Fileset. https://doi.org/10.6084/m9.figshare.7504448.v3 (2019). 11. Gatti, L. V. et al. Amazonia as a carbon source linked to
deforestation and climate change. Nature 595, 388–393
(2021). 32. Walsh, R. & Lawler, D. Rainfall seasonality: description, spatial pat-
terns and change through time. Weather 36, 201–208 (1981). 12. Lasslop, G., Brovkin, V., Reick, C. H., Bathiany, S. & Kloster, S. Mul-
tiple stable states of tree cover in a global land surface model due
to a fire-vegetation feedback. Geophys. Res. Lett. 43,
6324–6331 (2016). 33. Copernicus Climate Change Service (C3S). ERA5: Fifth generation
of ECMWF atmospheric reanalyses of the global climate. Coperni-
cus Climate Change Service Climate Data Store (CDS), https://cds. climate.copernicus.eu/cdsapp#!/home (2017). Nature Communications| (2023) 14:498 10 Article Funding 42. Hulme, M. & Kelly, M. Exploring the links between desertification
and climate change. Environ. Sci. Policy Sustain. Dev. 35,
4–45 (1993). g
Open Access funding enabled and organized by Projekt DEAL. Open Access funding enabled and organized by Projekt DEAL. Competing interests 43. Chen, J. et al. A simple method for reconstructing a high-quality
ndvi time-series data set based on the savitzky–golay filter. Remote
Sens. Environ. 91, 332–344 (2004). Competing interests
The authors declare no competing interests. Additional information
Supplementary information The online version contains
supplementary material available at
https://doi.org/10.1038/s41467-023-36207-7. 44. Cleveland, R. B., Cleveland, W. S., McRae, J. E. & Terpenning, I. Stl: A
seasonal-trend decomposition procedure based on loess. J. Off. Stat. 6, 3–73 (1990). 45. Smith, T. & Boers, N. Global Vegetation Resilience Linked to Water
Availability and Variability (1.0). Zenodo. https://doi.org/10.5281/
zenodo.7436669 (2023). Correspondence and requests for materials should be addressed to
Taylor Smith. 46. Friedl, M. & Sulla-Menashe, D. MCD12C1 MODIS/Terra+Aqua Land
Cover Type Yearly L3 Global 0.05Deg CMG V006. NASA EOSDIS
Land Processes DAAC. https://doi.org/10.5067/MODIS/MCD12C1. 006 (2015). Peer review information Nature Communications thanks Stephen Good
and the other, anonymous, reviewer(s) for their contribution to the peer
review of this work. Peer reviewer reports are available. Peer review information Nature Communications thanks Stephen Good
and the other, anonymous, reviewer(s) for their contribution to the peer
review of this work. Peer reviewer reports are available. 47. Gorelick, N. et al. Google earth engine: Planetary-scale
geospatial analysis for everyone. Remote Sens. Environ. 202,
18–27 (2017). Reprints and permissions information is available at
http://www.nature.com/reprints 48. Rousseau, D.-D. et al. (MIS3 & 2) millennial oscillations in Greenland
dust and Eurasian aeolian records - a paleosol perspective. Quat. Sci. Rev. 196, 99–113 (2017). Publisher’s note Springer Nature remains neutral with regard to jur-
isdictional claims in published maps and institutional affiliations. 49. Scheffer, M., Carpenter, S. R., Dakos, V. & van Nes, E. H. Generic
indicators of ecological resilience: inferring the chance of a critical
transition. Annu. Rev. Ecol. Evol. Syst. 46, 145–167 (2015). Open Access This article is licensed under a Creative Commons
Attribution 4.0 International License, which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as
long as you give appropriate credit to the original author(s) and the
source, provide a link to the Creative Commons license, and indicate if
changes were made. The images or other third party material in this
article are included in the article’s Creative Commons license, unless
indicated otherwise in a credit line to the material. If material is not
included in the article’s Creative Commons license and your intended
use is not permitted by statutory regulation or exceeds the permitted
use, you will need to obtain permission directly from the copyright
holder. To view a copy of this license, visit http://creativecommons.org/
licenses/by/4.0/. 50. Dakos, V. et al. Slowing down as an early warning signal for
abrupt climate change. Proc. Natl. Acad. Sci. USA 105,
14308–12 (2008). 51. Scheffer, M. et al. Early-warning signals for critical transitions. Nature 461, 53–9 (2009). 52. Boers, N. & Rypdal, M. Critical slowing down suggests that the
western greenland ice sheet is close to a tipping point. Proc. Natl. Acad. Sci. USA 118, e2024192118 (2021). 53. Boers, N. Observation-based early-warning signals for a collapse of
the atlantic meridional overturning circulation. Nat. Clim. Change
11, 680–688 (2021). 54. Djikstra, H. Nonlinear Climate Dynamics (Cambridge University
Press, 2013). © The Author(s) 2023 55. Kubo, R. The fluctuation-dissipation theorem. Rep. Prog. Phys. 29,
255–284 (1966). 11 Nature Communications| (2023) 14:498
|
https://openalex.org/W4288283499
|
https://zenodo.org/record/3332744/files/Suhartono.pdf
|
English
| null |
Effect of Pesticide on Human Health and Environment Maternal and Child Health Issues in Agricultural Areas in Brebes: Impact of Pesticide Use?
|
Zenodo (CERN European Organization for Nuclear Research)
| 2,019
|
cc-by
| 4,831
|
Effect of pesticide on human health and environment maternal and child
health issues in agricultural areas in Brebes: Impact of pesticide use? Suhartono
Environmental Health Department, Public Health Faculty, Diponegoro University PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 really need our attention together, because this phase is a golden period of
human life that will determine the quality of the nation in the future. So far,
efforts to overcome this problems have focused in the aspect of nutrition5,
because of the very strong relationship between nutrition and health and
growth6. The possible role of exposure to toxic substances in the environment
like pesticide as one of the main 'causes' of maternal and child health problems
also needs our attention. It is very potential as a risk factor for maternal and
child health problemsin agricultural areas. Even though it is not easy to ensure
that the high level of pesticide use is a main cause of various maternal and child
health problems in Brebes Regency, some findings from previous studies
supported by related theories demonstrated the harmful effects of pesticides on
maternal and childhealth. Research Results and Theories on the Impact of Pesticides on Maternal and
Child Health Theories and many previous studies on the effects of pesticides on
human health have been widely published but efforts to overcome this problem
have not been carried out optimally. The results of our studies regarding the
adverse effects of pesticides specifically on maternal and child health that had
been conducted in Brebes Regency and supported by theories and the results of
research abroad are presented in this paper. INTRODUCTION Brebes Regency located in the North Coast of Java Island (Pantai Utara
Jawa, Pantura) is the highest shallots production region in Indonesia. Shallots are
susceptible to pests like insects and fungi. Therefore, nearly all of the farmers in
this area mix various types of pesticides and exceed the recommended dose as
shown in a pesticide label. Data obtained from the Brebes Agriculture Office
demonstrated that the level of pesticide use in Brebes Regency was the largest
both in Indonesia and in Southeast Asia. There are approximately 3,200
pesticide brands registered by the Ministry of Agriculture of the Republic of
Indonesia in whichabout 1,300 of these brands are distributed in Brebes Regency
(Agriculture Office of Brebes Regency 2017). Our studiesconducted in several
villages in Brebes Regency showed that farmers in these area mixed 2-3 types of
pesticides before sprayingshallots with a frequency of spraying about 2-4 times
a week in dry season and every day in rainy season1. Amount of pesticide use
by shallot farmers in Brebes Regency was 330,000 liters per planting season in
which they had four planting seasonsa year (Agriculture Office of Brebes
Regency 2017). Some health problems, specifically related to Maternal and Child Health
(MCH), were widely found in Brebes Regency. Data from the Central Java
Province Health Office in 2016 showed that maternal mortality cases in Brebes
Regency were the highest in Central Java Province, reaching 54 cases. Meanwhile, infant mortality rates (13.42 per 1000 live births) placed the seventh
of 37 regencies/cities in Central Java2. The incidence of stunting in children in
Brebes Regency was also relatively high, reaching 40.7% 3. Stunting (height for
age < –2 SD of the WHO Child Growth Standards median), referred to as failure
of linear growthis a strong marker of unhealthy growth given its association
with morbidity and mortality risk, non-communicable diseases in later life, and
learning capacity and productivity. It is also closely linked with child
development in several domains including cognitive, language and sensory-
motor capacities4. The maternal mortality, infant mortality and stunting, are health
problems that often occur in the first 1000 days of life (from the beginning of the
fetus to two years old children). Health problems in the first 1000 days of life 93 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 Women with a history of exposure to pesticides are at risk three times to suffer
from hypothyroidism1. Hypothyroidism is a condition in which the body lacks
thyroid hormone which is characterized by increased levels of thyroid
stimulating hormone (TSH) and a decrease in T417. A study in Brazil showed a
link between pesticide exposure in the workplace and the incidence of thyroid
dysfunction (hypothyroid-like effects), especially in men18. Research in Iowa and
North Carolina (United States) demonstrated that exposure to chlordane
insecticides, benomyl and mancozeb fungicides, and paraquat herbicides were
risk factors for the incidence of hypothyroidism in women in agricultural
areas19. Benomyl and mancozeb, which are also widely used in the agricultural
area of Brebes district, are both fungicides with carbamate groups.The
dithiocarbamate pesticides ethylenebisdithiocarbamate and ethylenethiourea
have been associated with thyroid dysfunction. Maneb/mancozeb has been
shown to cause hypothyroidism in rabbits20, and human workers exposed to
ethylenethiourea showed significantly lower levels of total T4 compared with
unexposed controls21. In another study of unprotected workers heavily exposed
to ethylenebisdithiocarbamate, TSH levels were elevated, but T4 levels were no
different from those of controls22. Normal thyroid function in pregnant women is very important because
it will determine the quality of the child it contains23. Women with
hypothyroidism have decreased fertility; even if they conceive, risk of abortion
is increased, and risk of gestational hypertension, anemia, placentaabruption
and postpartum hemorrhage are increased24. Severe hypothyroidism has been
associated with pre-eclampsia, placental abnormalities, low birth weight infants,
and postpartum hemorrhage (bleeding)25. A cohort study in the Netherlands
provedthat maternal hypothyroxinaemia during early gestation was an
independent determinant of a delay in infant neurodevelopment. However,
when the thyroxin (fT4) concentrations increase during pregnancy in women
who are hypothyroxinaemic during early gestation, infant development appears
not to be adversely affected26. In Brebes Regency, women and children living in agricultural areas are
generally involved in various agricultural activities, such as finding pests and
removing shallots from their stalks ('mrotoli') after harvesting. These activities
are very risky to expose to pesticides because few days prior harvesting, shallot
farmers spray their shallot plants. Application of pesticides to crops may leave
residue on plants. Exposure to pesticides in humans can occur through several
routes, namely the respiratory tract (inhalation), direct contact with the skin or
through the gastrointestinal tract (by oral)27. Impact of Pesticides on the Thyroid Functions Some types of pesticides that are often used in the shallot farming area
of Brebes include Mancozeb, Dithane, and Chlorpyrifos7. These pesticides are
classified as endocrine disrupting chemicals (EDCs), chemical compounds in the
environment that can interfere with the biosynthesis, metabolism, and hormone
function in the body and can affect hormone balance function (homeostasis) and
reproduction8,9. A thyroid hormone is one of the hormone types that can be
disrupted by exposureto EDCs10,11. The hormone is indispensable in the process
of metabolism and human growth12,13. A previous study conducted in an
agricultural area in a sub district of Brebes Regency in 2010 demonstrated that
the prevalence of hypothyroidism among women of childbearing age reached
22.2% 14,higher than other cities in Indonesia like Jakarta, Bandung, Semarang,
Surabaya and Malang that was only 0.5%
15. Some studies conducted
internationally showed that the prevalence of hypothyroidism in women group
ranging from 0.3% to 2.9% 16. Our study also proves, that pesticide exposure is a risk factor for the
incidence of hypothyroidism in women of childbearing age1 and in children7. 94 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 The involvement of women and
children in agricultural activities increases the risk of exposure to pesticides,
especially exposure through direct contact with the skin. The 'mrotoli' activity is 95 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 usually carried out by them without using personal protective equipment like
masks or gloves to prevent pesticide residues in shallot plants entering their
bodies through respiratory air or skin pores. Pesticide contamination in the
environment (water, soil and food) also increases the risk of indirect exposure.A
study conducted by the Health Ministry's Health Research and Development
Agency of the Republic of Indonesia in several agricultural areas in Brebes
Regency demonstrated that five of 12 rivers or 42% were positively
contaminated with organophosphate pesticides residue28. In addition, the
Indonesian Agricultural Environment Research Institute reported that
chlorpyrifos pesticide residues were found in four of the five shallot samples
examined29. Our study in Brebes showed that pesticide exposure is a risk factor for
sub-clinical hypothyroidism (SCH) in school children living in an agricultural
areas30. SCH or mild hypothyroidism is characterized by mildly elevated serum
TSH concentrations, with normal concentrations of serum free and total
triiodothyronine (T3) and thyroxine (T4),without the typical symptoms of
thyroid disease31. The abnormalities most frequentlyassociated in the pediatric
population are goiter, poorschool performance, weight gain, increased
cholesterollevels, impaired growth velocity, anemia, excessivesleepiness,
weakness, and impaired psychomotor andcognitive development32,33.Another
study in Brebes Regency concluded that the proportion of goiter in the children
whose fathers were farmer (80.8%) were higher than those whose fathers non-
farmers (43.2%)34. The impact of pesticides on maternal health (pregnancy) The high maternal mortality rate in Brebes Regency might be related to
the high use of pesticides in the area. Data from the Brebes Regency Health
Office showed that the main cause of maternal mortality in Brebes was pre-
eclampsia (PE), which was 32% (Brebes District Health Office 2016). PE is an
increase in blood pressure (gestational hypertension) and proteinuria at over 20
weeks of pregnancy35.Hypertensive disorders of pregnancy are an important
cause of severe morbidity, long-term disability and death among both mothers
and their babies. In Africa and Asia, nearly one tenth of all maternal deaths are
associated with hypertensive disorders of pregnancy, whereas one quarter of
maternal deaths in Latin America have been associated with those
complications36. Several studies have shown that pesticide exposure increases
the risk of hypertension in pregnant women37,38.These studies suggest that the
risk of both Pregnancy Induced Hypertension (PIH) and PE was elevated among
women who performed activities likely to have exposed them to pesticides
during their first trimester of pregnancy37. A previous study reported of an 96 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 association between pesticide exposure and gestational diabetes39. Diabetes
caused endothelial dysfunction40, which may contribute to the pathophysiology
of PE41, as supported by studies showing that tight glycemic control reduces the
risk of PE in women who had diabetes before becoming pregnant42,43.Pesticide
exposure in pregnant women is also suspected of causing fetal growth disorder
and increase the risk of low birth weight. A studyin Brazilian rural areas
demonstrated that the per capita pesticide sales were directly associated with
higher prevalence of children born with low birth weight (r = 0.403), with birth
weights between 1500 and 2500 grams (r = 0.366), and very low birth weight
birth (r = 0.476). All correlations were statistically significant (p < 0.001)44. In addition to pesticides, other toxic substances that could increase the
risk of PE was lead(plumbum, Pb)45–47. Ikechukwu et al. (2012) stated that lead
increases the circulating levels of endothelin48. Endothelin is a vasoactive that
constricts the diameter of blood vessels thereby affecting blood pressure. Lead
also reduces serum levels of vasodilator substances such as nitric oxide (NO)
and endothelial-derived relaxation factor (EDRF) due to a lead-mediated
increase in reactive oxygen species49,50. Lead inhibits membrane adenosine
triphosphatases (ATPases), which increases intracellular calcium ions and
vasoconstriction51. The impact of pesticides on maternal health (pregnancy) Mechanisms cause preeclampsia either by inducing
vasoconstriction and placental ischemia or direct toxicity on the endothelial cell
and the renal function and thus causing proteinuria, which is a common feature
of preeclampsia48. The results of our study in six Puskesmas areas on the northern coast of
Brebes Regency showed that almost all pregnant women (98%) had blood Pb
levels above 10 µ / dL52, while the CDC threshold value was 5µg / dL53. This
study also provedpregnant women with high blood Pb levels were two times
more likely to develop hypertension (gestational hypertension) than those with
low blood Pb levels52.Some types of pesticides used in Brebes Regency that
contain lead were Antracol 70 WP, Buldok 25 EC dan Dithane M-45, Mancozeb
and Profenofos54. The impact of pesticides on children's growth and development Young children are particularly vulnerable to toxicants in the
environment, including pesticides55.Children’s organs are not fully developed
until later in life. They continually experience critical periods in development;
adverse exposures can cause permanent damage, particularly in utero56. Children’s behaviors and ability to interact with their physical environment
change during different stages of growth and development and can place them
at greater risk of exposure: children may crawl on the floor, explore objects
orally, and play with items they find in the environment57. For their weight, 97 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 children consume more food and drink than adults, increasing their possible
dietary exposure; dietary exposure is compounded by children’s immature
livers and excretory systems, which may be unable to effectively remove
pesticide metabolites57. These metabolites may block the absorption of critical
nutrients in children’s diets, which further affects health outcomes. Potential
routes of exposure to pesticides include in utero or through breast milk, via
ingestion of food contaminated with pesticides, and household exposures via
dermal contact56. Children live closer to the ground than adults, which may
increase their exposure to pesticides sprayed or precipitated there58. Research in 2014 in the Brebes agricultural area showed more than 30%
of primary school-aged children were detected dialkyl phosphates (DAPs),
metabolites of multiple organophosphorus pesticides, in their urine. Meanwhile,
the results of X-ray examination proved that 27 of the 50 children (54.0%) were
delay in skeletal maturation (bone-age delay)59. Another study by Kartini et alin
2017 proved that pesticide exposure was a risk factor for stuntingamong
primary school-aged children in Brebes agricultural areas, children with a
history of exposure to pesticides were 4.5 times risk of suffering from stunting
than those without exposure 60. Exposure to toxic substances in the environment, including pesticides,
leads to oxidative stress61 which led to an increase in the use of energy for the
activation of the immune system, thereby reducing the use of energy for the
maintenance, reproduction, growth and thermoregulation.62Exposure to
pesticides can also interfere with the function of several growth hormones, such
as IGF-1 hormone which is very necessary in the process of child growth63. Recent research suggests that even low levels of pesticide exposure can affect
young children’s neurological and behavioral development. Evidence shows a
link between pesticides and neonatal reflexes, psychomotor and mental
development, and attention-deficit hyperactivity disorder (ADHD). CONCLUSIONS People living in agricultural areas are at risk of being exposed to various
toxic substances, such as pesticides and heavy metals like lead (Pb) which are
contained in many pesticides. Exposure can occur directly, through their
involvement in agricultural activities, or indirectly through contact with the
environment contaminated by pesticide residues in water, air, soil, agricultural
or food products. Exposure to pesticides and leadwas a risk factor for various
types of health problems specifically related to hormonal dysfunction and the
quality of fetal or child growth and development. In Indonesia, distribution and
the use of pesticide needs to be well regulated in order to prevent adverse health
effects due to pesticide exposure in the future. PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 hyperactive behavior64,67. Children with levels higher than the median of
detectable concentrations had double the odds of ADHD (adjusted OR, 1.93
[95% CI, 1.23–3.02]) compared with those with non-detectable levels68. Leadcommonly contained in pesticides has been primarily classified as a
neurotoxin 69,70and is associated with developmental and behavioral problems in
children71–73. Preventing lead exposure in infants and young children is
important because lead can affect their developing brain and nervous system. High levels of lead can adversely affect the nervous system and kidneys of
adults and children74. A study in Bangladesh concluded that chronic lead
poisoning is significantly associated with high level of stunting among child
slum dwellers75. The impact of pesticides on children's growth and development In utero
dialkyl phosphate (DAPs), the organophosphate metabolites, and, to a lesser
extent, postnatal DAPs were associated adversely with attention as assessed by
maternal report, psychometrician observation, and direct assessment. These
associations were stronger at 5 years than at 3.5 years and were stronger in
boys64.A study in USA showed an adverse associations of prenatal DAPs with
mental development and pervasive developmental problems at 24 months of
age65. Another research concluded that organophosphate exposure might be
associated with deficits in learning on neurobehavioral performance,
particularly in tests of with motor function66.A study in the USA showed that
children who were detected organophosphate pesticide metabolites in their
urine were at risk of attention deficit hyperactivity disorder (ADHD), a
developmental disorder characterized by concentration difficulties and 98 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 REFERENCES Suhartono, Djokomoeljanto R, Hadisaputro S, Subagio H, Kartini A, Suratman. Pesticides exposure as a risk factor of hypothyroidism on childbearing
age women in the agricultural areas (in Bahasa). Media Med Indones. 2012;46(36):91-99. Health office of Central Java Province. Central Java Province Health Profile 2016. Semarang; 2017. Kementerian Kesehatan Republik Indonesia. Riset Kesehatan Dasar (Riskesdas)
2013.; 2013. World Health Organization. Global Nutrition Targets 2025: Stunting Policy Brief.;
2014. Kementerian Kesehatan Republik Indonesia. Kualitas Manusia Ditentukan Pada
1000 Hari Pertama Kehidupannya. 99 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 http://www.depkes.go.id/article/view/17012300003/kualitas-manusia-
ditentukan-pada-1000-hari-pertama-kehidupannya.html. Published
2017. Hurley KM, Yousafzai AK, Lopez-boo F. Early Child Development and
Nutrition : A Review of the Benefits and Challenges of Implementing
Integrated
Interventions. Adv
Nutr. 2016;7:357-363. doi:10.3945/an.115.010363. Hurley KM, Yousafzai AK, Lopez-boo F. Early Child Development and
Nutrition : A Review of the Benefits and Challenges of Implementing
Integrated
Interventions. Adv
Nutr. 2016;7:357-363. doi:10.3945/an.115.010363. Budiyono. Riwayat
paparan
pestisida
sebagai
faktor
risiko
kejadian
hipotiroidisme pada anak (studi pada anak sekolah dasar di daerah
pertanian
holtikultura
bawang
merah)
(Disertasi,
Universitas
Diponegoro). 2018. Diamanti-Kandarakis E, Bourguignon J, Giudice L, et al. Endocrine-Disrupting
Chemicals: An Endocrine Society Scientific Statement. Endocr Rev. 2009;30(4):293-342. doi:10.1210/er.2009-0002 Mnif W, Hassine A, Bouaziz A, Bartegi A, Thomas O, Roig B. Effect of endocrine
disruptor pesticides: A review. Int J Environ Res Public Health. 2011;8(6):2265-2303. doi:10.3390/ijerph8062265 Calsolaro V, Pasqualetti G, Niccolai F, Caraccio N, Monzani F. Thyroid
disrupting
chemicals. Int
J
Mol
Sci. 2017;18:1-17. doi:10.3390/ijms18122583 Miller M, Crofton K, Rice D, Zoeller R. Thyroid-disrupting chemicals:
Interpreting upstream biomarkers of adverse outcomes. Environ Health
Perspect. 2009;117(7):1033-1041. doi:10.1289/ehp.0800247 Mullur R, Liu Y-Y, Brent GA. Thyroid Hormone Regulation of Metabolism. Physiol Rev. 2014;94(2):355-382. doi:10.1152/physrev.00030.2013 Brent G. Tissue-Specific Actions of Thyroid Hormone : Insights From Animal
Models. Rev Endocr Metab Disord. 2000;1:27-33. Suhartono, Dharminto. Pesticide poisoning and hypothyroidism on childbearing
age women in agricultural areas [in Bahasa]. J Kesehat Masy Nas. 2010;4(5):217-222. Abbot Laboratories Diagnostic Division. Prevalence of Thyroid Disease in
Pregnancy in the South Asia Population (Unpublished).; 2008. Vanderpump M. The epidemiology of thyroid disease. Br Med Bull. 2011;99:39-
51. doi:10.1093/bmb/ldr030 American Tyhroid Association (ATA). Hypothyroidism. A Booklet for Patients and
Their Families.; 2013. www.thyroid.org. Piccoli C, Cremonese C, Koifman R, Koifman S, Freire C. REFERENCES Pesticide exposure and
thyroid function in an agricultural population in Brazil. Environ Res. 2016;151:389-398. doi:10.1016/j.envres.2016.08.011 100 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 Goldner W, Sandler D, Yu F, Hoppin J, Kamel F, Levan T. Pesticide use and
thyroid disease among women in the agricultural health study. Am J
Epidemiol. 2010;171(4):455-464. doi:10.1093/aje/kwp404 Mallem L, Boulakoud M, Franck M. Hypothyroidism after medium exposure to
the
fungicide
maneb
in
the
rabbit
Cuniculus
lepus. CommunAgricApplBiolSci. 2006;71(2a):91-99. Smith D. Ethylene thiourea: thyroid function in two groups of exposed workers. Br J Ind Med. 1984;41:362-366. Steenland K, Cedillo L, Tucker J, et al. Thyroid hormones and cytogenetic
outcomes in backpack sprayers using ethylenebis(dithiocarbamate)
(EBDC)
fungicides
in
Mexico. Environ
Health
Perspect. 1997;105(10):1126-1130. doi:10.1289/ehp.971051126 Lazarus J. Thyroid function in pregnancy. Br Med Bull. 2011;97:137-148. doi:10.1093/bmb/ldq039 Abalovich M, Gutierrez S, Alcaraz G, Maccallini G, Garcia A, Levalle O. Overt
and Subclinical Hypothyroidism Complicating Pregnancy. Thyroid. 2002;12(1):63-68. American Thyroid Association. Thyroid Disease and Pregnancy. American
Thyroid Association. www.thyroid.org. Published 2014. Pop V, Brouwers E, Vader H, Vulsma T, Van Baar A, De Vijlder J. Maternal
hypothyroxinaemia during early pregnancy and subsequent child
development: A 3-year follow-up study. Clin Endocrinol (Oxf). 2003;59(3):282-288. doi:10.1046/j.1365-2265.2003.01822.x\Kalayanova F,
El Batawi M. Human Toxicology of Pesticide. CRC Press; 1991. Badan Litbang Kemenkes RI. Riset Khusus Pencemaran Lingkungan [Laporan
Penelitian Tahun 2014].; 2014. Balai penelitian lingkungan pertanian. Residu Pestisida Di Bawang Merah.; 2018. Suhartono, Kartini A, Subagio H, et al. Pesticide exposure and thyroid function
in elementary school children living in an agricultural area, Brebes
District, Indonesia. Int J Occup Environ Med. 2018;9(3):137-144. doi:10.15171/ijoem.2018.1207 Biondi B, Cooper D. The clinical significance of subclinical thyroid dysfunction. Endocr Rev. 2008;29(1):76-131. doi:10.1210/er.2006-0043 Wu T, Flowers J, Tudiver F, Wilson J, Punyasavatsut N. Subclinical thyroid
disorders and cognitive performance among adolescents in the United
States. BMC Pediatr. 2006;6(12):1-6. doi:10.1186/1471-2431-6-12 Aijaz N, Flaherty E, Preston T, Bracken S, Lane A, Wilson T. Neurocognitive
function in children with compensated hypothyroidism: Lack of short
term effects on or off thyroxin. BMC Endocr Disord. 2006;6(2):1-7. doi:10.1186/1472-6823-6-2 101 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficienc
ISBN 978-602-344-251-5 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 Kartini A, Suhartono, Pangestuti D, Sakundarno M, Suratman, Rasipin. Goiter
and hypothyroidism among elementary school children in lowland
agricultural area, Brebes district Indonesia. Indian J Public Heal Res Dev. 2018;9(9):120-125. The American College of Obstetricians and Gynecologists. Hypertension in
Pregnancy.; 2013. World Health Organization. WHO Recommendations for Prevention and Treatment
of Pre-Eclampsia and Eclampsia. WHO Library Cataloguing-in-
Publication Data; 2011. Saldana T, Basso O, Baird D, et al. Pesticide Exposure and Hypertensive
Disorders During Pregnancy. Environ Health Perspect. 2009;117(9):1393-
1396. doi:10.1289/ehp.0900672 Ledda C, Fiore M, Santarelli L, et al. Gestational Hypertension and
Organophosphorus Pesticide Exposure : A Cross-Sectional Study. Biomed Res Int. 2015:5. doi:10.1155/2015/280891 Saldana T, Basso O, Hoppin J, et al. Pesticide exposure and self-reported
gestational diabetes mellitus in the agricultural health study. Diabetes
Care. 2007;30(3):529-534. doi:10.2337/dc06-1832 Dandona P, Aljada A, Chaudhuri A, Mohanty P. Endothilial Dysfunction in
Diabetes. In: Clinical Diabetes: Translating Research into Practice (Fonseca
V, Ed). Philadelphia: Saunders Elsevier; 2006. Lyell D, Lambert-Messerlian G, Giudice L. Prenatal screening, epidemiology,
diagnosis, and management of preeclampsia. Clin Lab Med. 2003;23(2):413-442. doi:10.1016/S0272-2712(03)00027-1 Crowther C, Hiller J, Moss J, McPhee A, Jeffries W, Robinson J. Effect of
Treatment of Gestational Diabetes Mellitus on Pregnancy Outcomes. N
Engl J Med. 2005;352(24):2477-2486. Fan Z, Yang H, Gao X, Lintu H, Sun W. Pregnancy outcome in gestational
diabetes. Int
J
Gynecol
Obstet. 2006;94(1):12-16. doi:10.1016/j.ijgo.2006.03.021 Boccolini P, Boccolini C, Meyer A, Chrisman J, Guimarães R, Veríssimo G. Pesticide exposure and low birth weight prevalence in Brazil. Int J Hyg
Environ Health. 2013;216:290-294. doi:10.1016/j.ijheh.2012.08.006 Bayat F, Akbari S, Dabirioskoei A, Nasiri M, Mellati A. The relationship between
blood lead level and preeclampsia. Electron Physician. 2016;8(12):3450-
3455. doi:http://dx.doi.org/10.19082/3450 Jameil N. Maternal serum lead levels and risk of preeclampsia in pregnant
women: A cohort study in a maternity hospital, Riyadh, Saudi Arabia. Int J Clin Exp Pathol. 2014;7(6):3182-3189. 102 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
ovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-
ISBN 978-602-344-251-5 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 Poropat A, Laidlaw M, Lanphear B, Ball A, Mielke H. Blood lead and
preeclampsia: A meta-analysis and review of implications. Environ Res. 2018;160:12-19. doi:10.1016/j.envres.2017.09.014 Ikechukwu I, Ojareva O, Ibhagbemien A, et al. Blood Lead, Calcium, and
Phosphorus in Women With Preeclampsia in Edo State, Nigeria. PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficienc
ISBN 978-602-344-251-5 Arch
Environ
Occup
Health. 2012;67(3):163-169. doi:10.1080/19338244.2011.619212 Carmignani M, Volpe A, Boscolo P, et al. Catcholamine and nitric oxide systems
as targets of chronic lead exposure in inducing selective functional
impairment. Life Sci. 2000;68(4):401-415. Gonick H, Behari J. Is lead exposure the principal cause of essential
hypertension? Med Hypotheses. 2002;59(3):239-246. doi:10.1016/S0306-
9877(02)00207-4 Moreau T, Hannaert P, Orssaud G, et al. Influence of membrane sodium
transport upon the relation between blood lead and blood pressure in a
general male population. Environ Health Perspect. 1988;78:47-51. doi:10.1289/ehp.887847 Suhartono, Kartini A, Budiyono, Darundiati Y. Paparan Pestisida Dan Plumbum
(Pb) Sebagai Faktor Risiko Kejadian Hipertensi Pada Kehamilan (Gestational
Hypertension): Studi Pada Ibu Hamil Di Daerah Pertanian Dan Pantai
Kabupaten Brebes. [Laporan Penelitian].; 2017. Ettinger A, Wengrovitz A. Guidelines for the Identification and Management of Lead
Exposure
in
Pregnant
and
Lactating
Women. Atlanta;
2010. https://www.cdc.gov/nceh/lead/publications/leadandpregnancy2010.p
df. Hartini E. Kadar plumbum (Pb) dalam darah pada wanita usia subur di daerah
pertanian. J Visikes. 2010;9(2). Liu J, Schelar E. Pesticide exposure and child neurodevelopment: summary and
implications. Work Heal Saf. 2012;60(5):235-243. doi:10.3928/21650799-
20120426-73. Chalupka S, Chalupka A. The impact of environmental and occupational
exposures on reproductive health. J Obstet Gynecol Neonatal Nurs. 2010;39(1):84-100. Landrigan P, Kimmel C, Correa A, Eskenazi B. Children’s health and the
environment: Public health issues and challenges for risk assessment. Environ Health Perspect. 2004;112(2):257-265. Paulson J, Barnett C. Who’s in Charge of Children’s Environmental Health at
School? New Solut. 2010;20(1):3-23. doi:10.2190/NS.20.1.b Kartini A, Suhartono, Subagio H, Budiyono, Emman I. Stunting and bone age
maturity in elementary school students in agriculture areas of Brebes 103 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
nnovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficien
ISBN 978-602-344-251-5 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 District
[in
Bahasa]. J
Kesehat
Masy. 2016;11(2):96-103. doi:http://dx.doi.org/10.15294/ kemas.v11i1.3462 Kartini A. Riwayat Paparan Pestisida Sebagai Faktor Risiko Kejadian Stunting Pada
Anak Sekolah Dasar Di Daerah Pertanian: Studi Di Kabupaten Brebes
[Disertasi].; 2017. Lukaszewicz-hussain A. Role of oxidative stress in organophosphate insecticide
toxicity – Short review. Pestic Biochem Physiol. 2010;98:145-150. doi:10.1016/j.pestbp.2010.07.006 Degen A. Effect of macroparasites on the energy budget of small mammals. In:
Morand S, Krasnov Y, Poulin R, eds. In: Morand S, Krasnov Y, Poulin
R, eds. Micromammals and Macroparasites: From Evolutionary Ecology to
Management. Tokyo: Springer-Verlag; 2006:371-400. Boada L, Lara P, Álvarez-León E, et al. Serum levels of insulin-like growth
factor-I in relation to organochlorine pesticides exposure. Growth Horm
IGF Res. 2007;17:506-511. doi:10.1016/j.ghir.2007.05.004 Marks A, Harley K, Bradman A, et al. Organophosphate pesticide exposure and
attention in young Mexican-American children: The CHAMACOS
study. Environ
Health
Perspect. 2010;118(12):1768-1774. doi:10.1289/ehp.1002056 Eskenazi B, Marks A, Bradman A, et al. Organophosphate pesticide exposure
and neurodevelopment in young Mexican-American children. Environ
Health Perspect. 2007;115(5):792-798. doi:10.1289/ehp.9828 Butler-Dawson J, Galvin K, Thorne P, Rohlman D. Organophosphorus pesticide
exposure and neurobehavioral performance in Latino children living in
an
orchard
community. Neurotoxicology. 2016;53:165-172. doi:10.1016/j.neuro.2016.01.009. Yu C, Du J, Chiou H, et al. Increased risk of attention-deficit/hyperactivity
disorder associated with exposure to organophosphate pesticide in
Taiwanese children. Andrology. 2016;4(4):695-705. Bouchard
M,
Bellinger
D,
Wright
R,
Weisskopf
M. Attention
deficit/hyperactivity
disorder
and
urinary
metabolites
of
organophosphate pesticides in U.S. children 8-15 years. Pediatrics. 2010;125(6):1-15. Lanphear B, Hornung R, Khoury J, et al. Low-level environmental lead exposure
and children’s intellectual function: An international pooled analysis. Environ Health Perspect. 2005;113(7):894-899. doi:10.1289/ehp.7688 p
p
Lidsky T, Schneider J. Lead neurotoxicity in children: Basic mechanisms and
clinical correlates. Brain. 2003;126(1):5-19. doi:10.1093/brain/awg014 dsky T, Schneider J. Lead neurotoxicity in children: Basic mechanisms and
clinical correlates. Brain. 2003;126(1):5-19. doi:10.1093/brain/awg014 104 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 PROCEEDING OF INTERNATIONAL WORKSHOP AND SEMINAR
Innovation of Environmental-Friendly Agricultural Technology Supporting Sustainable Food Self-Sufficiency
ISBN 978-602-344-251-5 Lewendon G, Kinra S, Nelder R, Cronin T. Should children with developmental
and behavioural problems be routinely screened for lead? Arch Dis
Child. 2001;85:286-288. Gould E. Childhood lead poisoning: Conservative estimates of the social and
economic benefits of lead hazard control. Environ Health Perspect. 2009;117(7):1162-1167. doi:10.1289/ehp.0800408 Wisconsin Department of Health Services. Lead Arsenate Pesticides in Soil. Human Health Hazards. 2010. Wisconsin Department of Health Services. Lead Arsenate Pesticides in Soil. Human Health Hazards. 2010. Raihan M, Briskin E, Mahfuz M, et al. Examining the relationship between blood
lead level and stunting, wasting and underweight- A cross-sectional
study of children under 2 years-of-age in a Bangladeshi slum. PLoS
One. 2018;13(5):1-15. doi:10.1371/journal.pone.0197856 Raihan M, Briskin E, Mahfuz M, et al. Examining the relationship between blood
lead level and stunting, wasting and underweight- A cross-sectional
study of children under 2 years-of-age in a Bangladeshi slum. PLoS
One. 2018;13(5):1-15. doi:10.1371/journal.pone.0197856 105
|
https://openalex.org/W1862648057
|
https://library.oapen.org/bitstream/20.500.12657/35261/1/340069.pdf
|
Dutch
| null |
Honderdvijftig jaar levenslopen : De Historische Steekproef Nederlandse bevolking
| null | 2,008
|
cc-by
| 98,935
|
Ineke Maas, Marco H.D. van Leeuwen en Kees Mandemakers (red.) Ineke Maas, Marco H.D. van Leeuwen en Kees Mandemakers (red.) Ineke Maas, Marco H.D. van Leeuwen en Kees Mandemakers (red.) Ineke Maas, Marco H.D. van Leeuwen en Kees Mandemakers (red.) De Historische Steekproef
Nederlandse bevolking
Honderdvijftig
jaar levenslopen Honderdvijftig jaar levenslopen Honderdvijftig jaar levenslopen Onder redactie van Ineke Maas
Marco H.D. van Leeuwen
Kees Mandemakers De Historische Steekproef Nederlandse bevolking Onder redactie van
Ineke Maas
Marco H.D. van Leeuwen
Kees Mandemakers Honderdvijftig jaar levenslopen: de Historische Steekproef Nederlandse bevolking is de
boekaflevering bij jaargang 83 (2008) van het sociaal-wetenschappelijk tijdschrift
Mens & Maatschappij. Omslagafbeelding: Volendam, juli 1959
Foto: Ben van Meerendonk / AHF, collectie IISG, Amsterdam Omslagafbeelding: Volendam, juli 1959
Foto: Ben van Meerendonk / AHF, collectie IISG, Amsterdam Omslagontwerp: Jos Hendrix, Groningen
Vormgeving binnenwerk: ProGrafici, Goes isbn 978 90 8964 067 3
e-isbn 978 90 4850 662 0
nur 741 © Ineke Maas, Marco H.D. van Leeuwen en Kees Mandemakers / Amsterdam Uni-
versity Press, Amsterdam 2008 © Ineke Maas, Marco H.D. van Leeuwen en Kees Mandemakers / Amsterdam Uni-
versity Press, Amsterdam 2008 Alle rechten voorbehouden. Niets uit deze uitgave mag worden verveelvoudigd, opgeslagen in een
geautomatiseerd gegevensbestand, of openbaar gemaakt, in enige vorm of op enige wijze, hetzij
elektronisch, mechanisch, door fotokopieën, opnamen of enige andere manier, zonder voorafgaande
schriftelijke toestemming van de uitgever. Voorzover het maken van kopieën uit deze uitgave is toegestaan op grond van artikel 16B Auteurswet
1912 jº het Besluit van 20 juni 1974, Stb. 351, zoals gewijzigd bij het Besluit van 23 augustus 1985,
Stb. 471 en artikel 17 Auteurswet 1912, dient men de daarvoor wettelijk verschuldigde vergoedin-
gen te voldoen aan de Stichting Reprorecht (Postbus 3051, 2130 KB Hoofddorp). Voor het overnemen
van gedeelte(n) uit deze uitgave in bloemlezingen, readers en andere compilatiewerken (artikel 16
Auteurswet 1912) dient men zich tot de uitgever te wenden. Inhoudsopgave Ineke Maas, Marco H.D. van Leeuwen en Kees Mandemakers
7
Honderdvijftig jaar levenslopen: een inleiding
Kees Mandemakers
De database van de Historische Steekproef Nederlandse bevolking (HSN)
9
Hilde Bras, Aart C. Liefbroer en Cees H. Elzinga
Standaardisering van leefvormen? Trajecten naar volwassenheid
15
van Nederlanders, 1850-1940
Jan Van Bavel, Jan Kok en Theo Engelen
Hoge kinderloosheid tijdens het interbellum in Nederland. 51
De rol van godsdienst, levensstandaard en economische crisis
Matthijs Kalmijn
Voorlopers in de echtscheidingsrevolutie. De relatie tussen
81
echtscheiding en sociale klasse in de negentiende en
vroeg-twintigste eeuw
Hans Knippenberg en Sjoerd de Vos
Vroege ontkerkelijking in Nederland. Een analyse van het
97
geboortecohort 1850-1882
Onno Boonstra
Functioneel analfabetisme in Nederland, 1775-1900
127
Richard L. Zijdeman en Kees Mandemakers
De rol van het gymnasiaal en middelbaar onderwijs bij de
149
intergenerationele overdracht van status, Nederland 1865-1940 Ineke Maas, Marco H.D. van Leeuwen en Kees Mandemakers
Honderdvijftig jaar levenslopen: een inleiding Ineke Maas en Marco H.D. van Leeuwen
Van een dubbeltje naar een kwartje? Beroepsloopbanen
173
van mannen en vrouwen in Nederland tussen 1865 en 1940
Frans van Poppel en Ruben van Gaalen
Sociale klasse, sociale mobiliteit en sterfte in Nederland, 1850-2007
203
Over de auteurs
237 6 Over de auteurs Honderdvijftig jaar levenslopen: een inleiding Ineke Maas, Marco H.D. van Leeuwen en Kees Mandemakers Veranderingen in de wijze waarop mensen hun leven vormgeven zijn soms heel
abrupt – denk aan de gevolgen van de verdwijning van het IJzeren Gordijn voor de
levenslopen van Midden- en Oost-Europeanen. Vaker echter veranderen de levens-
lopen van mensen langzaam, van de ene generatie op de andere. Grootouders
herkennen zich nog in hun kinderen, maar nauwelijks meer in hun kleinkinde-
ren. Voor kleinkinderen is het onvoorstelbaar hoe hun grootouders leefden. Hoewel we allemaal verhalen kennen over ‘hoe het vroeger was’, was er tot
voor kort weinig systematische kennis over levenslopen van mensen geboren voor
1900. Grootschalig sociologisch onderzoek naar levenslopen begon in Nederland
in de jaren negentig van de twintigste eeuw (Ultee & Ganzeboom, 1995). Ook
oudere mensen werden geïnterviewd over hun levensloop en dat leverde gege-
vens op voor de periode vanaf 1900 (zie bijvoorbeeld Liefbroer & Dykstra, 2000). Historisch onderzoek naar levenslopen reikt verder terug, maar betrof ofwel
hele levenslopen van slechts een klein aantal personen, of alleen een bepaald
aspect van de levensloop: bijvoorbeeld het beroep op het moment van huwen. Veel vragen over veranderingen op de lange termijn konden daarom tot nu toe
niet worden beantwoord. In 1987 startte een samenwerking tussen onderzoekers uit de historische en
sociale wetenschappen, gericht op het verzamelen van levensloopdata van een
omvangrijke representatieve steekproef uit de Nederlandse bevolking geboren
tussen 1812 en 1922: het project Historische Steekproef Nederlandse bevolking
(HSN).1 Het project wordt uitgevoerd door het Internationaal Instituut voor Soci-
ale Geschiedenis (IISG) te Amsterdam. In eerste instantie richtte de dataverza-
meling zich op de akten van de burgerlijke stand die werden opgemaakt bij de
meest markante momenten in de levens van individuen: geboorte, huwelijk en
overlijden.2 Vanaf 2003 maakte een NWO-subsidie (Programma voor grote inves-
teringen nr. 175-107.105.01) het mogelijk om in het project Life Courses in Con-
text ook dynamische informatie te verzamelen. Deze is afkomstig uit het Neder- landse bevolkingsregister dat vanaf 1849 werd bijgehouden. Hierin wordt voor
iedereen de woonplaats, de gezinssituatie, het beroep en de godsdienst geno-
teerd en regelmatig bijgewerkt, bijvoorbeeld bij een verhuizing, een geboorte of
bij de tienjaarlijkse volkstelling. 8 De eerste uit dit project afkomstige dataset is onlangs – in 2007 – beschikbaar
gekomen voor onderzoekers. Dit boek bevat een eerste overzicht van onderzoek
naar levenslopen van Nederlanders geboren tussen 1850 en 1922. Honderdvijftig jaar levenslopen: een inleiding Het multidis-
ciplinaire karakter van de onderzoeksgroep is zichtbaar zowel in de auteurs van
de hoofdstukken als in de aspecten van de levensloop die aan de orde komen. Sociologen, historici, demografen, geografen en statistici bestuderen trajecten
naar volwassenheid, kinderloosheid, echtscheiding, ontkerkelijking, analfabe-
tisme, beroepsmobiliteit en sterfte. De hoofdstukken hebben in de eerste plaats een beschrijvende waarde: ze laten
zien hoe levenslopen vanaf 1850 eruitzagen en welke verschillen er waren tussen
sociale groepen. Daarnaast worden in alle hoofdstukken bestaande theorieën en
hypothesen getoetst. Deze theorieën hebben specifiek betrekking op de geboorte-
cohorten van voor 1900 ofwel ze gaan over langzame veranderingen gedurende de
afgelopen twee eeuwen. Voor het eerst worden ze nu getoetst met gegevens over
levenslopen die ook werkelijk op deze periode betrekking hebben. Met de verzamelde data kunnen nog veel meer vragen worden beantwoord
en andere hypothesen worden getoetst. De dataset staat ter beschikking voor
geïnteresseerde onderzoekers.3 Daarnaast zijn er vergevorderde plannen om
nieuwe databronnen aan te boren die nog uitgebreidere informatie opleveren
over de onderzoekspersonen in het databestand, zoals bijvoorbeeld over hun
familienetwerk of over militaire dienst. Dit boek is daarom niet de afsluiting van
het onderzoek naar historische levenslopen, maar juist het begin van de studie
naar de levens van onze ouders, grootouders en overgrootouders. Noten 1. Zie http://www.iisg.nl/~hsn/. p //
g
/
/
2. Deze dataverzameling werd mogelijk gemaakt door een reeks subsidies van het ministerie
van Onderwijs & Wetenschappen, NWO en de KNAW. Zie voor de details het HSN Jaarver-
slag 2007. 2. Deze dataverzameling werd mogelijk gemaakt door een reeks subsidies van het ministerie
van Onderwijs & Wetenschappen, NWO en de KNAW. Zie voor de details het HSN Jaarver-
slag 2007. 3. Zie http://www.iisg.nl/~hsn/. 3. Zie http://www.iisg.nl/~hsn/. Literatuur HSN (2007). Jaarverslag 2007. Amsterdam: IISG. Liefbroer, A.C. & P.A. Dykstra (2000). Levenslopen in verandering. Een studie naar ont-
wikkelingen in de levenslopen van Nederlanders geboren tussen 1900 en 1970. Den
Haag: Sdu Uitgevers. Liefbroer, A.C. & P.A. Dykstra (2000). Levenslopen in verandering. Een studie naar ont-
wikkelingen in de levenslopen van Nederlanders geboren tussen 1900 en 1970. Den
Haag: Sdu Uitgevers. Ultee, W.C. & H.B.G. Ganzeboom (1995). Netherlands family survey 1992-93. Nijme-
gen: Department of Sociology, Nijmegen University. Ultee, W.C. & H.B.G. Ganzeboom (1995). Netherlands family survey 1992-93. Nijme-
gen: Department of Sociology, Nijmegen University. 3. Zie http://www.iisg.nl/~hsn/. De database van de Historische Steekproef
Nederlandse bevolking (HSN) Kees Mandemakers Het doel van de Historische Steekproef Nederlandse bevolking (HSN) is de recon-
structie van de gehele levensloop van een groot aantal personen geboren in de
periode voordat survey-onderzoek zijn intrede deed. De onderzoekspersonen
voor de HSN-database zijn geselecteerd door middel van een aselecte steekproef
uit de geboorteregisters uit de periode 1812-1922. De steekproeffractie varieert
per geboorteperiode: 0,75 procent voor de periode 1812-1872, 0,5 procent voor de
periode 1873-1902 en 0,25 procent voor de periode 1903-1922. Daarnaast zijn de
provincies Zeeland, Friesland en Utrecht voor de periode 1903-1922 overtrokken
tot een totaal van 0,5 procent. Het exacte aantal steekproefpersonen bedraagt
78.105. Dit is iets meer dan een half procent van de in totaal 14,5 miljoen per-
sonen die in deze periode in Nederland zijn geboren. Een aantal van 78.105 is
voldoende om statistisch verantwoorde uitspraken te kunnen doen voor relatief
kleine subpopulaties binnen de Nederlandse bevolking (Mandemakers, 2000). Voor uitgebreide informatie over de HSN kan worden verwezen naar de website:
www.iisg.nl\~hsn. Buiten Nederland kent men al geruime tijd vergelijkbare projecten, vooral in
Canada, Zweden, Engeland, Noorwegen, België, Frankrijk en de Verenigde Staten
(Kelly Hall, McCaa & Thorvaldsen, 2000). De HSN onderscheidt zich doordat het
onderzoek zich niet over bepaalde regio’s maar over het gehele land uitstrekt, en
bovendien migranten gevolgd zijn naar hun nieuwe woonplaatsen waardoor ook
voor hen de gehele levensloop kon worden vastgelegd. In deze zin is de HSN-database
uniek te noemen. De HSN gaat dus uit van een onderzoeksconcept waarbij Neder-
land als één gebied en de daarin bewaard gebleven bevolkingsregisters als één bron
worden beschouwd, ondanks de geografische verspreiding en optredende plaatse-
lijke verschillen in de kwaliteit en toegankelijkheid van dit onderzoeksmateriaal. In de database van de HSN worden voor al deze 78.105 onderzoekspersonen
op systematische wijze gegevens verzameld uit de burgerlijke stand (allereerst de
geboorteakten, daarna overlijdensakten en huwelijksakten, zie verder Vulsma, 2002). Met de geboorteakten is niet alleen de basis gelegd voor het onderzoek
naar de levenslopen. In de akten zelf vinden we meteen al een groot aantal gege-
vens, zoals de namen, adressen, leeftijden en beroepen van de ouders. Tevens
zien we of de aangever (meestal de vader) zijn handtekening kon zetten of niet. Bij de tot op heden in de database opgenomen overlijdensakten ligt de nadruk
op de overlijdensakten van vroeg overleden kinderen. De database van de Historische Steekproef
Nederlandse bevolking (HSN) Bij de oudere personen
kan in de regel pas een akte worden gevonden als de gehele levensloop bekend
is.1 Overleden kinderen werden meestal aangegeven door de vader, zodat voor
vaders een tweede mogelijkheid ontstaat voor het noteren van het uitgeoefende
beroep en het al dan niet zetten van een handtekening, indicator van analfabe-
tisme. Huwelijksakten zijn een zeer rijke bron. Ze bevatten namelijk niet alleen
gegevens van de huwenden, maar ook van de ouders van het bruidspaar en van
twee of vier getuigen. Dit zijn veelal vrienden of familie van de huwenden. De
akte bevat dus diverse aan elkaar gerelateerde personen met daarbij voor alle
personen gegevens over woonplaats, leeftijd, beroep, handtekening en relatie. 10 10 Het uiteindelijke doel van de HSN is de reconstructie van de gehele levens-
loop van de in de steekproef opgenomen personen. Hiervoor wordt ook alle infor-
matie die in de bevolkingsregisters is te vinden, overgenomen. Door koppeling
van alle bevolkingsregisters van de meer dan duizend toen bestaande gemeen-
ten wordt het mogelijk het volledige migratiepatroon van al deze personen uit
te zoeken. Ook blijkt uit deze bron de kerkelijke gezindte en opnieuw vaak het
beroep. De gezinssamenstelling komt naar voren uit de relatie die de verschillen-
de personen in het huishouden ten opzichte van het hoofd van het gezin inne-
men, bijvoorbeeld ‘zoon’, ‘nicht’ of ‘kostganger’. Op basis van de migratiegege-
vens en de gegevens betreffende de geboortedatum en eventuele sterfdatum kan
voor elk moment in de tijd de samenstelling van het gezin en de veranderingen
daarin worden bepaald (Knotter en Meier, 1995; Mandemakers, 2006a). Het overnemen van de gegevens uit de registers gebeurt niet alleen voor de
onderzoekspersonen, maar ook voor alle andere personen die zich in hetzelfde
gezin bevinden. Een typische levensloop kenmerkt zich door vier verschillende
situaties: a) opgroeien binnen het ouderlijke gezin, b) inwonen bij andere gezin-
nen of in kosthuissituaties als dienstbode, schoolleerling, ambachtsleerling,
kostganger, dienstplichtige of anderszins, c) als ouders met een eigen gezin en
d) inwonen als bejaarde of behoeftige. Zeker gezien de hoge kindersterfte in het
verleden maakte lang niet elke onderzoekspersoon alle fasen mee. En ook niet
iedereen kwam aan een huwelijk toe, kreeg kinderen, of had voor het huwelijk
het ouderlijke huis reeds verlaten. Het bevolkingsregister ging eind jaren dertig, officieel per 1 januari 1940,
over van de gezinskaarten op de persoonsgezinskaarten (PK). De database van de Historische Steekproef
Nederlandse bevolking (HSN) Als een persoon van
de ene gemeente naar een andere verhuisde, dan verhuisde deze PK mee. Dit
systeem is blijven werken tot 1 oktober 1994, toen de overgang naar de Gemeen-
telijke Basis Administratie (GBA) werd gerealiseerd. In het geval van overlijden
werd de PK gearchiveerd bij het Centraal Bureau voor Genealogie. Van alle onder- kees mandemakers zoekspersonen die op 1 januari 1940 nog in leven waren, werden de PK’s door
de HSN opgevraagd en indien gevonden ingevoerd. Ook de PK is rijk aan infor-
matie; zo zijn onder andere de beroepsvermeldingen, het gehele migratietraject
(alle adressen), de gezinssamenstelling en de godsdienstige gezindte te vinden. Deze unieke bron maakt het mogelijk om tot ver in de twintigste eeuw door te
gaan met onderzoek naar onderwerpen als sociale mobiliteit, ontkerkelijking,
veranderingen in de gezinssamenstelling en migratiepatronen. Samen met de
overlijdensakten is nu voor meer dan de helft van alle HSN-onderzoekspersonen
bekend waar en op welk moment ze zijn overleden. 11 Samenstelling van de HSN dataset Levenslopen, release 2007.01 De verzameling van de levenslopen van alle personen in de HSN-steekproef is
nog niet afgesloten. Een belangrijke stap is gezet binnen het kader van het door
NWO-Groot en de KNAW gesubsidieerde project Life Courses in Context (Mande-
makers, 2004). Binnen dit project zijn voor 40.000 personen, iets meer dan de
helft van de oorspronkelijke steekproef, de levenslopen gevolgd en ingevoerd. Het betreft onderzoekspersonen die geboren zijn tussen 1850 en 1922. Het twee-
de deel van dit project, de context, digitaliseert de volkstellingen zoals die zijn
gepubliceerd tussen 1859 en 1947. Dit onderdeel werd uitgevoerd door het Neder-
lands Instituut voor Wetenschappelijke Informatiediensten (NIWI) en later door
Data Archiving and Networking Services (DANS). Het gaat hierbij om samenge-
stelde gegevens op het niveau van de gemeente of van de provincie, zoals de
leeftijdsopbouw of samenstelling van de beroepsbevolking. Voor deze gegevens
zie www.volkstellingen.nl. Dit onderdeel werd 29 september 2006 afgerond door
middel van het symposium Uitgeteld en ingevoerd: analyse van de Nederlandse volkstel-
lingen 1795-2001 (Boonstra e.a., 2007). Met het congres Levenslopen in de 19e en 20e
eeuw werd op vrijdag 11 april 2008 het gehele Life Courses in Context-programma
afgesloten. Een deel van de hoofdstukken in dit boek werden gepresenteerd op
dit congres. Andere bijdragen aan dit congres zullen in 2009 verschijnen in een
themanummer van het Tijdschrift voor Sociale en Economische Geschiedenis. De in dit boek opgenomen hoofdstukken zijn gebaseerd op de tussentijdse
dataset Historische Steekproef Nederlandse bevolking (HSN), dataset Levenslopen, release
2007.01. Deze dataset wordt in dit boek verder aangehaald als HSN dataset Levens-
lopen, release 2007.01. Voor de wijze waarop de brongegevens tot levenslopen zijn
verwerkt, zie Mandemakers (2006b). De HSN dataset Levenslopen, release 2007.01 kwam uit in januari 2007. Op dat
moment bevatte de HSN-dataset de levenslopen van 19.535 onderzoekspersonen. Van 452 personen werden geen gegevens in het bevolkingsregister gevonden door
slechte indexen of bijvoorbeeld doordat er geen registers meer waren. Een voor-
beeld hiervan zijn de registers van Middelburg die voor de periode tot 1900 zijn
verbrand tijdens de Tweede Wereldoorlog. Het effectieve aantal onderzoeksper-
sonen komt daardoor op 19.083 (zie tabel 1). Voor het maken van deze dataset, de database van de historische steekproef nederlandse bevolking (hsn) werden er prioriteiten bij de dataverzameling gesteld. Deze prioritering leverde
een steekproef op die bestaat uit twee onderdelen. 1. Samenstelling van de HSN dataset Levenslopen, release 2007.01 Personen geboren in de periode 1850-1922 voor de provincies Friesland, Zee-
land, Utrecht en de stad Rotterdam (n = 12.972). Deze provincies maakten al
eerder deel uit van HSN-onderzoek naar levenslopen. Deze voorsprong was
reden om er met verhoogde prioriteit verder aan te werken. 12 2. Personen geboren in de periode 1883-1922 in de rest van Nederland. Daarbij
werd voor de periode 1883-1902 ad random 60 procent van de oorspronkelijke
steekproef geselecteerd, dit is dus een effectieve steekproef van 0,3 % (0,6*0,5). Samen met de 0,25 % van de periode 1903-1922 resulteert dit in 13.817 perso-
nen. Tabel 1 Samenstelling HSN dataset Levenslopen, release 2007.01
Provincie Friesland, Utrecht
Zeeland en stad Rotterdama
Rest
Nederland
Totaal
1850-1882
1883-1922
1883-1922
1850-1922
Aantal
%
Aantal
%
Aantal
%
Aantal
%
Totaal aantal onderzoekspersonen
6333
100,0
6639
100,0
13817
100,0
26789
100,0
Nog niet opgenomen in dataset
926
14,6
938
14,1
5390
39,0
7254
27,1
Niet gevonden in bevolkings-
register
257
4,1
64
1,0
131
0,9
452
1,7
In de dataset opgenomen levens
lopen
5150
81,3
5637
84,9
8296
60,0
19083
71,2
Daarvan volledig gevolgd
3999
63,1
4553
68,6
7702
55,7
16254
85,2
Daarvan onvolledig gevolgd
1151
18,2
1084
16,3
594
4,3
2829
14,8
a Rotterdam omvat de geannexeerde gemeenten Delfshaven, Hillegersberg, Schiebroek, Overschie,
Katendrecht, Charlois, Kralingen, Pernis, Hoogvliet, IJsselmonde en Poortugaal. Tabel 1 Samenstelling HSN dataset Levenslopen, release 2007.01 a Rotterdam omvat de geannexeerde gemeenten Delfshaven, Hillegersberg, Schiebroek, Overschie,
Katendrecht, Charlois, Kralingen, Pernis, Hoogvliet, IJsselmonde en Poortugaal. De mate van volledigheid varieert per provincie en periode. De dataset voor
Utrecht en Zeeland is nagenoeg compleet en die van Friesland en Rotterdam
komen uit op ongeveer 75 %. Bij elkaar gaat het om 85 % van de in totaal 12.972
in aanmerking komende onderzoekspersonen. Van de rest van het land uit de
geboorteperiode 1883-1922, komt de volledigheid uit op 60 % van de in aanmer-
king komende onderzoekspersonen. Het ontbreken van levenslopen is niet at
random, het gaat vooral om personen die of heel veel verhuisden of naar buiten
de geboorteregio zijn verhuisd. Er is dus een duidelijke oververtegenwoordiging
van ‘blijvers’. Samenstelling van de HSN dataset Levenslopen, release 2007.01 Ook kon niet van alle personen de volledige levensloop worden
gevonden en ingevoerd.2 Ten behoeve van de hoofdstukken in dit boek werd in 2007 ook een tussen-
tijdse release van de burgerlijke stand uitgebracht: Historische Steekproef Neder- kees mandemakers landse bevolking (HSN), dataset Akten Burgerlijke Stand, bèta-release 2007.01, hier ver-
der aangehaald als HSN release akten 2007.01. Huwelijksgegevens zijn met name
benut in de hier opgenomen hoofdstukken van Kalmijn en Boonstra. De release
bevat gegevens van geboorteakten (78.105), huwelijksakten (22.419) en overlijden
(40.637, waarvan 22.482 overlijdensakten en 18.155 persoonskaarten). Scheidings-
gegevens werden apart uitgegeven als HSN release akten_h_scheiding 2007.01. 13 Noten 1. Bovendien wordt er voor overlijdensakten niet verder gezocht dan 1 januari 1940, omdat
vanaf dat moment de persoonskaarten die bij het Centraal Bureau voor Genealogie berus-
ten, gebruikt worden. 1. Bovendien wordt er voor overlijdensakten niet verder gezocht dan 1 januari 1940, omdat
vanaf dat moment de persoonskaarten die bij het Centraal Bureau voor Genealogie berus-
ten, gebruikt worden. 2. Bij het verschijnen van deze publicatie is de dataset Historische Steekproef Nederlandse bevol-
king (HSN), dataset Levenslopen, release 2008.01 uitgekomen. In deze dataset zijn enkele klei-
ne fouten gecorrigeerd en is het aantal levenslopen uitgebreid tot meer dan 22.000. 2. Bij het verschijnen van deze publicatie is de dataset Historische Steekproef Nederlandse bevol-
king (HSN), dataset Levenslopen, release 2008.01 uitgekomen. In deze dataset zijn enkele klei-
ne fouten gecorrigeerd en is het aantal levenslopen uitgebreid tot meer dan 22.000. 2. Bij het verschijnen van deze publicatie is de dataset Historische Steekproef Nederlandse bevol-
king (HSN), dataset Levenslopen, release 2008.01 uitgekomen. In deze dataset zijn enkele klei-
ne fouten gecorrigeerd en is het aantal levenslopen uitgebreid tot meer dan 22.000. Standaardisering van leefvormen?
Trajecten naar volwassenheid van Nederlanders, 1850-1940 Hilde Bras, Aart C. Liefbroer en Cees H. Elzinga Hilde Bras, Aart C. Liefbroer en Cees H. Elzinga Hilde Bras, Aart C. Liefbroer en Cees H. Elzinga Literatuur Boonstra, O.W.A., P.K. Doorn, M.P.M. van Horik, J.G.S.J. van Maarseveen & J. Oud-
hof (red.) (2007). Twee eeuwen Nederland geteld. Onderzoek met de digitale Volks-
Beroeps- en Woningtellingen 1795-2001. Den Haag: DANS en CBS. Kelly Hall, P., R. McCaa & G. Thorvaldsen (red.) (2000). Handbook of International
Historical Microdata for Population Research. Minneapolis: Minnesota Population
Center. Knotter, A. & A.C. Meijer (red.) (1995). De gemeentelijke bevolkingsregisters 1850-19 Knotter, A. & A.C. Meijer (red.) (1995). De gemeentelijke bevolkingsregisters 1850-1920. Den Haag: Instituut voor Nederlandse Geschiedenis. Den Haag: Instituut voor Nederlandse Geschiedenis. Mandemakers, K. (2000). Historical Sample of the Netherlands. In P. Kelly Hall, R. McCaa & G. Thorvaldsen (red.), Handbook of International Historical Microdata for
Population Research (pp. 149-178). Minneapolis: Minnesota Population Center. Mandemakers, K. (2004). De Historische Steekproef Nederlandse bevolking (HSN)
en het project Life Courses in Context. Bevolking en Gezin, 33, 91-114. Mandemakers, K. (2006a). Levenslooponderzoek in Rotterdam met de Historische
Steekproef Nederlandse bevolking (HSN). In P. van de Laar, L. Lucassen & K. Mandemakers (red.), Naar Rotterdam. Immigratie en levensloop in Rotterdam vanaf
het einde van de negentiende eeuw (pp. 9-24). Amsterdam: Aksant. Mandemakers, K. (2006b). Building life course datasets from population registers
by the Historical Sample of the Netherlands (HSN). History and Computing, 14,
87-108. Mandemakers, K. (2006b). Building life course datasets from population registers
by the Historical Sample of the Netherlands (HSN). History and Computing, 14,
87-108. Vulsma, R.F. (2002). Burgerlijke stand en bevolkingsregister. 2e herziene druk. Den
Haag: Centraal Bureau voor Genealogie. Vulsma, R.F. (2002). Burgerlijke stand en bevolkingsregister. 2e herziene druk. Den
Haag: Centraal Bureau voor Genealogie. lsma, R.F. (2002). Burgerlijke stand en bevolkingsregister. 2e herziene druk. Den Vulsma, R.F. (2002). Burgerlijke stand en bevolkingsregister. 2e herziene druk. Den
Haag: Centraal Bureau voor Genealogie. de database van de historische steekproef nederlandse bevolking (hsn) Inleiding Cornelia Meeuwisse wordt geboren in 1921 in Hellevoetsluis. Vlak na de oorlog,
op 24-jarige leeftijd, trouwt ze met Jan Mak en betrekken ze een klein huisje in
Den Helder. Een jaar later wordt hun eerste kind geboren. Daarna volgen er nog
tien andere kinderen (Mak, 2005). Cornelia’s levensloop voldoet aan wat sociolo-
gen de standaardlevensloop noemen: uit huis gaan om te trouwen, kort daarna
gevolgd door de geboorte van het eerste kind (Liefbroer & De Jong Gierveld, 1993;
Van Leeuwen, 1987). Moeder thuis, vader verdient de kost. Het is het beeld van
de jaren vijftig en zestig. Hoe anders verliep het leven van Pietje Willemse, die
ruim zestig jaar eerder werd geboren in het Zeeuwse Tholen. Op 20-jarige leeftijd
verlaat zij het ouderlijk huis om als dienstmeisje te gaan werken; eerst in Zierik-
zee, daarna bij verschillende werkgevers in Amsterdam. Pietje laat haar periode
als dienstbode pas achter zich als zij op 26-jarige leeftijd trouwt en in Haarlem
bij haar schoonouders intrekt. Zij overlijdt echter al op 39-jarige leeftijd en laat
daarbij vier kinderen achter (Blikman-Ruiterkamp, 2000). Ook de overgang naar
de volwassenheid van Annetje Beets detoneert met het beeld van de standaard-
levensloop. Annetje wordt in 1888 in Purmerend geboren als dochter van een
besteller. Ze volgt een verpleegstersopleiding, wordt kraamverpleegster en later
huishoudster en woont afwisselend in zusterhuizen, bij familie en bij werkge-
vers. Annetje blijft tot haar 55ste vrijgezel (Oort, 2006). Levensloopsociologen spreken van ‘standaardisering’ om het proces aan
te duiden waarbij de levensloop eenvormiger wordt voor grotere delen van de
bevolking. De oorsprong van dit proces ligt volgens hen niet alleen bij demogra-
fische veranderingen en toegenomen welvaart, maar ook bij het ontstaan van
leeftijdsnormen die verbonden zijn met de expansie van het onderwijssysteem,
de groei van de moderne verzorgingsstaat en het socialeverzekeringswezen. In
de loop van de twintigste eeuw zou de met deze instituties verbonden leeftijds-
gerelateerde wetgeving de levensloop steeds meer zijn gaan structureren (Mayer, 1986). Standaardisering houdt daarom in de eerste plaats een proces van tempo-
ralisering in; een ontwikkeling waarbij leeftijd verandert van een categorische
status naar een structurerend element in het bestaan (Hagestad, 1992; Hagestad
& Neugarten, 1985; Kohli, 1986). Daarnaast zou er sprake zijn van chronologise-
ring waarbij gebeurtenissen in de levensloop steeds meer in een vaste volgorde
en op nauwe door normen bepaalde leeftijden plaatsvonden, resulterende in de
zogenaamde ‘gestandaardiseerde normatieve levensloop’ (Baars, 1991). Inleiding 16 16 In sociologisch onderzoek worden de flexibele ‘keuzelevenslopen’ uit het
huidige tijdsgewricht vaak afgezet tegen de standaardlevensloop uit ‘het verle-
den’. Maar was die standaardlevensloop wel zo representatief voor de ervarin-
gen van eerdere cohorten? Of gaat het hierbij slechts om de gemeenschappelijke
ervaringen van enkele generaties, zoals wel is gesuggereerd (Van Eijk, 2000)? Amerikaans onderzoek op basis van historische volkstellingen laat zien dat de
overgang naar de volwassenheid nog een extreem uitgerekte en ongeordende
fase was voor personen die in het midden van de negentiende eeuw geboren wer-
den. De fase werd pas geleidelijk aan uniformer voor generaties die geboren wer-
den vanaf het begin van de twintigste eeuw (Hogan, 1981; Modell, Furstenberg &
Hershberg, 1976; Modell, Furstenberg & Strong, 1978; Stevens, 1990). Ook in een
aantal Nederlandse studies werd een trend naar standaardisering waargenomen
voor cohorten geboren tussen 1903 en 1940 (Liefbroer & De Jong Gierveld, 1993;
Liefbroer & Dykstra, 2000). Aan de hand van surveys en interviews, waarop deze
studies zijn gebaseerd, kan echter niet verder teruggegaan worden dan tot cohor-
ten geboren in het begin van de twintigste eeuw. Hoe uniform waren levens-
lopen eigenlijk in de negentiende eeuw? Waar lagen precies de wortels van het
standaardiseringsproces? Het meeste levenslooponderzoek heeft zich beziggehouden met afzon-
derlijke levensloopgebeurtenissen. De determinanten van de frequentie, duur
en timing van transities, zoals trouwen en uit huis gaan, zijn daarbij in kaart
gebracht. In deze studie richten wij ons op ‘holistische’ trajecten zoals die tot
uitdrukking komen in de aaneenschakeling van gebeurtenissen gedurende een
bepaalde levensfase en/of op een bepaald levensdomein. Een dergelijk traject-
perspectief biedt een waardevolle aanvulling op het gangbare onderzoek naar
standaardiseringprocessen. Sociale veranderingen beïnvloeden immers niet
alleen afzonderlijke transities, maar ook de volgorde en timing daarvan binnen
complexe ketens van gedragskeuzen. Bovendien combineren trajecten oorzaak
en gevolg en reflecteren zo op een andere manier dan transities de geleefde wer-
kelijkheid (Hynes & Clarkberg, 2005; Pollock, 2007). We bestuderen in deze bij-
drage trajecten tijdens de overgangsfase naar de volwassenheid (15-40 jaar) en
richten ons in het bijzonder op trajecten van leefvormen. Leefvormen betreffen
de huishoudsamenstellingen en woonarrangementen waarin individuen zich
bevinden, zoals wonen met ouders, bij een werkgever verblijven, met partner en
kinderen samenleven of met drie generaties wonen. We onderzoeken allereerst of er sprake was van een proces van standaardise- hilde bras, aart c. liefbroer en cees h. Inleiding elzinga ring van leefvormtrajecten in de periode 1850-1940. In hoeverre werden trajecten
uniformer, hoe zagen karakteristieke trajecten eruit en in hoeverre werd een
bepaald pad naar de volwassenheid in de loop van de tijd algemener? In de twee-
de plaats trachten we groepsverschillen in het proces van standaardisering te
achterhalen. In hoeverre verschilden de leefvormtrajecten van mannen en vrou-
wen? In het debat over de modernisering van demografisch gedrag in Nederland
spelen sociale klasse en religie een belangrijke rol. We toetsen daarom ook speci-
fieke hypothesen betreffende sociale en religieuze verschillen in trajecten naar
de volwassenheid. 17 Aan de hand van de gegevens uit de Historische Steekproef Nederland (HSN)
bestuderen we leefvormcarrières van Nederlanders geboren tussen 1850 en 1900. De HSN is een database met historische registratiegegevens, waaronder gege-
vens uit de bevolkingsregisters en informatie uit de akten van de burgerlijke
stand (geboorte, huwelijk en overlijden). Deze dataset heeft een aantal belang-
rijke voordelen: ten eerste verschaft het longitudinale gegevens op individueel
niveau. De schaarse studies waarin (historische) trajecten zijn bestudeerd heb-
ben over het algemeen een synthetische cohortbenadering toegepast op basis
van volkstellingsgegevens (Glick, 1947; Glick & Parke, 1965; Modell, Furstenberg
& Hershberg, 1976; Stevens, 1990; Uhlenberg, 1969, 1974). Ondanks de waarde-
volle informatie die zulke herhaalde cross-sectionele surveys opleveren, kunnen
alleen longitudinale data de volgorde van keuzen en transities die individuen
doorlopen, inzichtelijk maken (Mouw, 2005). Ten tweede is de HSN-database
prospectief; informatie betreft niet alleen ‘overlevers’, maar ook individuen die
relatief vroeg overlijden. Op basis van de HSN kan een vollediger beeld geschetst
worden van de diversiteit aan levensloopervaringen van historische cohorten. In dit hoofdstuk onderzoeken we trajecten naar volwassenheid aan de hand
van methoden van sequentieanalyse. Sequentieanalyse maakt het mogelijk om,
door exploratie van patronen van similariteit en afstand, verschillende typen
levensloopervaringen te onderscheiden. We doen dit met een door Elzinga (2005)
ontwikkelde methode. Het hoofdstuk is als volgt opgezet. In de volgende paragraaf ontwikkelen we
hypothesen over algemene trends in de standaardisering van leefvormtrajecten
in Nederland en in het bijzonder over verschillen tussen mannen en vrouwen,
sociale klassen en kerkelijke gezindten. Deze hypothesen worden vervolgens
getoetst met de HSN-data. Eerst bezien we of er zich in het algemeen een pro-
ces van standaardisering heeft voorgedaan. Met behulp van clusteranalyse zijn
daarna uit de data verschillende typen trajecten gegenereerd. We onderzoeken
cohortverschillen in de gevolgde trajecten. Veranderingen in de jongvolwassenheid en de standaardiserings
hypothese Industrialisering en de jongvolwassenheid 18 De levensfase van jongvolwassenheid kent verschillende belangrijke transities en
rites de passage, zoals uit huis gaan, inwonen, trouwen en ouderschap. Daarnaast
is de periode van jongvolwassenheid een historisch veranderlijk verschijnsel; de
vorm, duur en inhoud ervan zijn nauw verweven met bredere maatschappelijke
ontwikkelingen (Brinkgreve & De Regt, 1991). Om hypothesen met betrekking tot
standaardisering van deze levensfase te formuleren, is het daarom nodig eerst
inzicht te krijgen in de belangrijkste historische veranderingen in patronen van
uit huis gaan, migratie, samenwonen met familie of niet-verwanten, huwelijks-
sluiting, ouderschap, vruchtbaarheid en sterfte. In de preïndustriële samenleving was de jongvolwassenheid een langgerek-
te, semi-autonome periode met een eigen sociale status; jongeren vormden een
aparte groep in de samenleving met eigen rituelen, feesten en verenigingen. De
fase kende nauwelijks leeftijdsgradaties; jongelui, van welke leeftijd dan ook,
bevonden zich in een vaag gebied tussen afhankelijkheid en onafhankelijkheid
(Gillis, 1974; Hanawalt, 1992). Het begin van de jongvolwassenheid werd gemar-
keerd door biologische rijping, religieuze initiatie en het beëindigen van het
lager onderwijs. Een huwelijk en het stichten van een eigen huishouden werden
over het algemeen gezien als de voltooiing ervan. De uitgerekte fase van jong-
volwassenheid was nauw verbonden met het heersende patroon van huwelijk
en voortplanting. In Nederland gold, net als in de rest van West-Europa, vanaf
ongeveer 1600 het West-Europese (of malthusiaanse) huwelijkspatroon (Hajnal,
1965). Dit patroon, dat door de Nederlandse demograaf Hofstee (1981) het agra-
risch-ambachtelijke patroon is genoemd, werd gekenmerkt door een hoge tot
zeer hoge huwelijksleeftijd, een relatief groot aandeel nooit huwenden en een
bijna maximale huwelijksvruchtbaarheid. Trouwen was alleen mogelijk wan-
neer men in staat was om zich van een bestaan te verzekeren dat representatief
was voor de eigen klasse of stand. Binnen een agrarisch-ambachtelijke samenle-
ving bracht een aanzienlijk deel van de jongeren een periode in het huishouden
van anderen door als gezel, leerling, meid of knecht. Het in het Engels zo tref-
fend aangeduide ‘life-cycle service’ (Laslett, 1977) bood jongeren de mogelijkheid
om ervaring op te doen met de werkzaamheden in een bepaalde beroepsgroep
en te sparen voor een uitzet of het opbouwen van een startkapitaal. Ook onder
de arbeiders in de steden zou volgens Hofstee dit patroon hebben bestaan. In het agrarisch-ambachtelijke patroon kwam verandering tijdens de peri-
ode van industrialisering. Die kwam in Nederland halverwege de jaren zestig
van de negentiende eeuw op gang (Van Zanden & Van Riel, 2004). Inleiding Aan de hand van een serie van logisti-
sche-regressieanalyses bekijken we in hoeverre er verschillen naar sekse, sociale
klasse en religie bestaan in de trajecten die individuen op weg naar de volwas-
senheid doorlopen. De belangrijkste resultaten en de implicaties daarvan bedis-
cussiëren we in de laatste paragraaf. standaardisering van leefvormen? Veranderingen in de jongvolwassenheid en de standaardiserings
hypothese Nederland ver-
schilde vooral van andere Europese landen doordat de industrialisering geken-
merkt werd door een intensivering van de dienstverlenende sector in plaats van
door voornamelijk een groei van de (primaire) industriële productie (Wintle, hilde bras, aart c. liefbroer en cees h. elzinga 2000). Hoewel Nederland al vanaf de zestiende eeuw een relatief geürbaniseerd
land was en vooral het westen sterk gecommercialiseerd was, ging de industria-
lisering gepaard met verdergaande urbanisatie. Werkgelegenheid en bevolkings-
groei namen eerst vooral in de grote steden toe, terwijl in een latere fase ook
de provinciesteden zich ontwikkelden (Diederiks, 1992). Er ontstond een scala
aan nieuwe beroepsmogelijkheden. Voor mannen nam het werk in de havens,
scheepsbouw, fabrieken en woningbouw toe. Tegelijkertijd steeg de vraag naar
vrouwelijk inwonend dienstpersoneel bij de groeiende middenklasse in de ste-
den. Deze uitbreiding van de werkgelegenheid ging gepaard met toegenomen
migratie, met name van het platteland naar de steden. Industrialisering en urba-
nisatie leidden ook tot de vorming van duidelijk afgebakende klassen. Er vorm-
den zich een stedelijke industriële arbeidersklasse, een klasse van industriële
en commerciële ondernemers en een nieuwe middenklasse van employés in het
bedrijfsleven en ‘witte boorden’ werkzaam in de groeiende overheidsbureaucra-
tie (De Regt, 1993a). 19 19 Door de stijging van de reële lonen en de algemene welvaartsverhoging
was het voor jongeren gemakkelijker geworden om al op jongere leeftijd econo-
mische zelfstandigheid te bereiken. Tijdens de laatste decennia van de negen-
tiende eeuw daalde de huwelijksleeftijd van jongeren dan ook geleidelijk. Deze
daling deed zich in alle sociale lagen voor (Van Poppel, 1992b). Er werd echter
niet alleen jonger maar ook meer getrouwd. Het aandeel celibatairen op 40- tot
44-jarige leeftijd daalde vanaf 1900 voor mannen, terwijl het voor vrouwen eerst
nog toenam, en pas vanaf 1910 geleidelijk en na 1950 snel verminderde (Engelen
& Kok, 2003). Ook een aantal andere belangrijke demografische veranderingen
vonden in deze periode plaats. Verbeteringen in economische omstandigheden,
gezondheidszorg, persoonlijke hygiëne en zuigelingenzorg hadden vanaf 1875
een snelle daling van de sterfte tot gevolg, wat niet alleen een verlenging van de
levensduur en een drastische reductie van de kindersterfte betekende, maar ook
een vermindering van de sterfte op jongvolwassen leeftijd (Van Poppel, 1999). Vanaf 1879 ging ook de huwelijksvruchtbaarheid dalen. Veranderingen in de jongvolwassenheid en de standaardiserings
hypothese Het bevolkingsaantal
werd niet meer – zoals binnen het agrarisch-ambachtelijke patroon gebruike-
lijk was – door laat of nooit trouwen onder controle gehouden, maar het aantal
geboortes werd nu binnen het huwelijk beperkt. Er waren echter grote sociale,
religieuze en regionale verschillen in de Nederlandse vruchtbaarheidstransitie
(Van Poppel, 1974, 1985). Industrialisering ging maar tot op zekere hoogte gepaard met een uitbrei-
ding van staatsorganisaties en wetgeving. De arbeidswetgeving bleef beperkt tot
het aan banden leggen van de kinderarbeid (in 1874 en 1889) en de regulering
van de arbeid van gehuwde vrouwen. Veel van het werk dat door jongeren werd
verricht, bijvoorbeeld in de huishoudelijke dienstensector, was echter van die
verordeningen uitgesloten. Ook het onderwijssysteem ging niet direct de jong-
volwassenheid bepalen. In 1905 werd de leerplicht ingevoerd en die was zesjarig,
tot en met gemiddeld twaalf jaar. In 1951 was dit slechts met twee jaar verlengd standaardisering van leefvormen? en waren jongeren tot en met gemiddeld hun veertiende jaar leerplichtig (Man-
demakers, 1996). Evenmin kwam de verzorgingsstaat in Nederland snel van de
grond. De armenzorg werd traditioneel zowel door de kerken als door de bur-
gerlijke armbesturen verzorgd, en dit bleef zo gedurende de gehele negentiende
eeuw (Van Leeuwen, 1994). Tijdens de tweede helft van de negentiende eeuw nam
de rol van de kerken af en gingen de gemeenten relatief steeds meer geven. Pas
in 1963 met de Algemene Bijstandswet werd de armenzorg een taak van de lan-
delijke overheid (Van der Valk, 1986). Wetgeving omtrent ouderdomspensioenen
werd in 1919 effectief, terwijl de Ziektewet niet eerder dan in 1930, en andere
sociale wetgeving zelfs pas na de Tweede Wereldoorlog werd geïmplementeerd
(Wintle, 2000). Van standaardisering door overheidsingrijpen was voor de Twee-
de Wereldoorlog dus nog nauwelijks sprake. 20 20 Technologische modernisering, ontwikkeling van de infrastructuur en van
nieuwe transportmogelijkheden (trein, tram en fiets) tijdens de laatste helft van
de negentiende eeuw en de eerste drie decennia van de twintigste eeuw drukten
wel een belangrijke stempel op de jongvolwassenheid (Knippenberg & De Pater,
1988; Van der Woud, 2007). Jongeren konden hierdoor vaker bij hun ouders thuis
blijven wonen en naar hun werk pendelen. Door de toegenomen welvaart en
de verruimde beroepsmogelijkheden en doordat men meer vrije tijd en privacy
wenste, nam de noodzaak en de behoefte ook af om als kostganger, dienstbode of
knecht in de huishoudens van anderen te wonen en te werken. Veranderingen in de jongvolwassenheid en de standaardiserings
hypothese Tijdens de periode 1850-1940 gingen jongeren vaker en vroeger trouwen,
werd de sterfte op jongvolwassen leeftijd geringer en woonden zij minder vaak in
bij niet-verwanten. Al deze ontwikkelingen beïnvloedden de jongvolwassenheid
op ingrijpende wijze. Verwacht kan worden dat er zich daardoor voor cohorten
geboren tussen 1850 en 1900 een proces van standaardisering van leefvormtra-
jecten naar de volwassenheid voltrok. Standaardisering van leefvormtrajecten Standaardisering van leefvormtrajecten Brückner en Mayer (2005) hebben concepten ontwikkeld om diverse aspecten van
standaardisatieprocessen in levenslopen van elkaar te onderscheiden. De concep-
ten verwijzen naar verschillende dimensies van standaardisering die onafhanke-
lijk van elkaar kunnen variëren. Een hiervan is het concept van de-differentiatie
(of homogenisering) dat het proces samenvat waarbij voorheen gesplitste levens-
fasen samenkomen (Brückner & Mayer, 2005). Als er sprake is van homogenise-
ring neemt het aantal verschillende stadia of posities in leefvormtrajecten af. De
afname van bijvoorbeeld in de negentiende eeuw gebruikelijke leefvormen zoals
inwonen als kostganger, in een internaat, of bij verwanten en het daarvoor in
de plaats langer bij de ouders thuis blijven wonen voor het huwelijk is een voor-
beeld van een dergelijk homogeniseringproces. Niet alleen de afname van het aantal stadia is relevant, ook het toevallige
karakter daarvan speelt een rol. Oudere cohorten hadden wellicht vaker grillige hilde bras, aart c. liefbroer en cees h. elzinga patronen met episodes van afwisselend wonen bij ouders, grootouders, werk-
gevers, familieleden of kostgezinnen dan jongere cohorten. Volgens Elzinga en
Liefbroer (2007) dekt het begrip turbulentie het vluchtige en toevallige karakter
van een dergelijke zeer gedifferentieerde levensloop. Dit leidt ons tot onze eerste
hypothese. 21 H1 De turbulentie in trajecten naar volwassenheid is voor cohorten geboren tussen 1850 en
1900 afgenomen. H1 De turbulentie in trajecten naar volwassenheid is voor cohorten geboren tussen 1850 en
1900 afgenomen. Naast homogenisering kan standaardisering als dimensie worden onderschei-
den. Het begrip standaardisering verwijst volgens Brückner en Mayer (2005) naar
het proces waarbij specifieke posities en de volgorde waarin ze voorkomen alge-
mener worden voor een bepaalde populatie. Standaardisering bestaat in feite uit
twee aspecten die weer apart onderscheiden kunnen worden: het steeds meer
gelijk worden van levenslopen en het afnemen van de diversiteit aan levensloop-
paden (Elzinga & Liefbroer, 2007). Het eerste aspect van standaardisering is dat
levenslopen meer op elkaar zijn gaan lijken. Dit leidt tot onze tweede hypothe-
se. H2 Trajecten naar volwassenheid van cohorten geboren tussen 1850 en 1900 zijn meer op
elkaar gaan lijken. H2 Trajecten naar volwassenheid van cohorten geboren tussen 1850 en 1900 zijn meer op
elkaar gaan lijken. Een tweede aspect van standaardisering is afname in de diversiteit aan trajecten
en toename van de dominantie van een specifiek traject. Peter Uhlenberg (1974)
ontwikkelde, op basis van censusgegevens over de levenslopen van Amerikaanse
vrouwen die geboren werden tussen 1890 en 1934, een typologie van vijf trajec-
ten. Het traject vroege dood werd gevolgd door vrouwen die tot hun vijftiende jaar
in leven bleven, maar voor hun vijftigste stierven. Vrouwen die op hun vijftig-
ste nog niet gehuwd waren, vielen in de categorie vrijgezellen. Vrouwen die wel
trouwden maar geen kinderen kregen, werden als kinderloos getypeerd. Vrouwen
die huwden en kinderen kregen, maar wiens eerste huwelijk door verweduwing
of scheiding uit elkaar viel, doorliepen het traject gebroken huwelijk met kinderen. Ten slotte onderscheidt Uhlenberg een traject dat door huwelijk, ouderschap en
het gezamenlijk opvoeden van de kinderen getypeerd wordt. Dit traject, waarin
gezinsvorming centraal staat, wordt door Uhlenberg het geprefereerde of stan-
daardtraject genoemd. Onder de door hem bestudeerde cohorten vrouwen kwam
dit traject relatief steeds vaker voor. Op basis van Uhlenbergs conclusies nemen we aan dat een dergelijk stan-
daardtraject, waarin gezinsvorming centraal stond, ook geleidelijk aan domi-
nanter werd voor Nederlandse cohorten geboren tijdens de tweede helft van de
negentiende eeuw. Onze derde hypothese is aldus geformuleerd. standaardisering van leefvormen? H3 De diversiteit van trajecten naar volwassenheid van cohorten geboren tussen 1850 en
1900 is afgenomen en de dominantie van een ‘standaardtraject’ getypeerd door gezins
vorming is toegenomen. H3 De diversiteit van trajecten naar volwassenheid van cohorten geboren tussen 1850 en
1900 is afgenomen en de dominantie van een ‘standaardtraject’ getypeerd door gezins
vorming is toegenomen. 22 22 Tot nog toe hebben we over processen van homogenisering en standaardisering
gesproken alsof die zich voor iedereen in dezelfde mate en in gelijk tempo voor-
deden. In het demografisch gedrag van Nederlanders in deze periode beston-
den echter grote verschillen naar sekse, sociale klasse en religieuze gezindte. Ten eerste valt het te verwachten dat de modernisering van de jongvolwassen-
heid bij mannen sneller haar beslag kreeg dan bij vrouwen. We hebben al gezien
dat de afname van het vrijgezellenbestaan bij mannen eerder plaatsvond dan
bij vrouwen. Vanaf 1909 begon het aandeel gehuwde mannen te stijgen voor-
dat dezelfde ontwikkeling zich voordeed bij vrouwen. In 1930 lag de proportie
nooit huwenden bij mannen nog steeds zo’n vijf procent lager dan bij vrouwen
(Engelen & Kok, 2003). H2 Trajecten naar volwassenheid van cohorten geboren tussen 1850 en 1900 zijn meer op
elkaar gaan lijken. Ook gingen jongens vaker en langer naar school dan meis-
jes (Knippenberg & De Pater, 1988). De belangrijkste oorzaak van verschillen in
uniformiteit in de jongvolwassenheid waren echter seksespecifieke arbeidsmo-
gelijkheden; jonge mannen konden veel gemakkelijker in het ouderlijk huis
blijven wonen en thuis of buitenshuis werken. Voor Nederlandse vrouwen was
het dienstbodeberoep van oudsher een belangrijke optie en de relevantie van dit
beroep als manier om de jongvolwassenheid te overbruggen nam alleen maar
toe gedurende de tweede helft van de negentiende eeuw. Alhoewel ook mannen
bij werkgevers en familie hun intrek namen om deel te nemen aan de stedelijk-
industriële arbeidsmarkt, kwam deze leefvorm relatief vaker op het levenspad
van vrouwen, waardoor hun jongvolwassenheid een grilliger en minder uniform
karakter kreeg. Dit brengt ons tot onze vierde hypothese. H4 Standaardisering van leefvormtrajecten naar volwassenheid vond in sterkere mate plaats
bij mannen dan bij vrouwen. H4 Standaardisering van leefvormtrajecten naar volwassenheid vond in sterkere mate plaats
bij mannen dan bij vrouwen. Ook tussen de sociale klassen bestonden van oudsher grote verschillen in patro-
nen van uit huis gaan, huwelijkssluiting, sterfte en vruchtbaarheid. Jongeren uit
de ongeschoolde arbeidersklasse werden veel vaker dan kinderen uit de boeren-
stand of uit de middenklassen knecht, meid of dienstbode (Bras, 2002). De daling
van de huwelijksleeftijd en van het celibaat onder invloed van de welvaartstoe-
name was echter veel pregnanter voor jongeren uit de arbeidersklasse dan voor
kinderen afkomstig uit boerenmilieus en de burgerij (Van Poppel, 1992b). Jonge-
ren uit de arbeidersklasse, die door middel van loonarbeid aan de kost moesten
zien te komen, profiteerden van de grotere vraag naar arbeid en de gestegen
lonen. Alhoewel de middenklasse vooropliep bij het toepassen van geboorte-
beperking (Van Bavel & Kok, 2005), betekende dit niet dat zij ook voortrekkers
waren in het vroege huwen. De materiële eisen voor het stichten van een eigen
huishouden waren door de toename van de levensstandaard alleen maar hoger hilde bras, aart c. liefbroer en cees h. elzinga geworden, waardoor het huwelijk nog moeilijker haalbaar werd. Velen gingen
daardoor geen relatie aan (Falkenburg, 1905). In een studie op basis van individu-
ele gegevens uit de persoonskaarten van cohorten geboren tussen 1890 en 1909
vonden Engelen en Kok (2003) dat jongeren, en vooral vrouwen, afkomstig uit
de hoogste sociale klassen, niet alleen later trouwden maar ook relatief vaker
ongehuwd bleven. H2 Trajecten naar volwassenheid van cohorten geboren tussen 1850 en 1900 zijn meer op
elkaar gaan lijken. Het West-Europese huwelijkspatroon bleef ook bij boeren nog
lang het gebruikelijke patroon; bezit speelde daar een belangrijke rol. Alhoewel
arbeidersjongeren vaker inwoonden bij werkgevers en hogere sterftekansen had-
den (Van Poppel, 1999), zullen vooral de toegenomen mogelijkheden om te trou-
wen en om dat op jongere leeftijd te doen ervoor gezorgd hebben dat hun jonge
levens uiteindelijk homogener werden, meer op elkaar gingen lijken en vaker
het standaardtraject benaderden dan dat bij jongeren uit andere sociale klassen
het geval was. Dit brengt ons tot de volgende hypothese. 23 H5 Standaardisering van leefvormtrajecten naar volwassenheid vond in sterkere mate bij
jongeren uit de arbeidersklasse plaats dan bij jongeren uit andere sociale klassen. Ten slotte verwachten we religieuze verschillen in de standaardisering van de
jongvolwassenheid. Katholieke geestelijken verwierpen tot ver in de twintigste
eeuw geboortebeperking binnen het huwelijk. Daardoor was het traditionele
patroon van huwelijksrestrictie (laat en niet trouwen) de enige manier van bevol-
kingscontrole. Binnen sommige delen van het protestantisme daarentegen werd
het neomalthusiaanse alternatief al veel eerder geaccepteerd. Vooral remon-
stranten en mennonieten waren vrijdenkers op het gebied van de geboortebeper-
king. Gereformeerden en andere orthodoxe protestanten hadden daarentegen
eenzelfde strenge seksuele moraal als de katholieken (Engelen & Kok, 2003; Van
Bavel & Kok, 2005). Ook was er een verschil in sterftekansen tussen katholieken
en protestanten; katholieken zouden door hun ongezondere levensstijl hogere
sterftecijfers hebben gehad (Van Poppel, 1992a). In vergelijking met katholieken
en orthodoxe protestanten zou onder vrijzinnig-protestanten dus eerder een
proces van standaardisering van de jongvolwassenheid plaats hebben gevonden. Onze zesde hypothese luidt aldus. H6 Standaardisering van leefvormtrajecten naar volwassenheid vond in sterkere mate plaats
bij jongeren uit vrijzinnig-protestantse gezinnen dan bij jongeren van andere religieuze
gezindten. H6 Standaardisering van leefvormtrajecten naar volwassenheid vond in sterkere mate plaats
bij jongeren uit vrijzinnig-protestantse gezinnen dan bij jongeren van andere religieuze
gezindten. Onderzoeksopzet Constructie van leefvormtrajecten De onderzoeksvragen worden beantwoord met behulp van gegevens uit de Histo-
rische Steekproef Nederlandse bevolking (HSN). De HSN is een nationaal databe- standaardisering van leefvormen? standaardisering van leefvormen? stand waarin informatie is opgenomen over de levensloop van een groot aantal
personen geboren in Nederland in de jaren 1812-1922 (Mandemakers, 2000). We
analyseren data van de Historische Steekproef Nederlandse bevolking (HSN), dataset
Levenslopen, release 2007.01. In deze dataset is alle informatie over de onderzoeks-
personen uit de bevolkingsregisters opgeslagen, zie ook de bijdrage van Mande-
makers in dit boek. 24 24 De dataset bevat de levenslopen van 19.535 onderzoekspersonen. De dataset
is gestratificeerd naar regio en periode. Voor de provincies Friesland, Zeeland,
Utrecht en de stad Rotterdam zijn levenslopen beschikbaar voor de geboorteco-
horten 1850-1922 (n = 10.787), voor de overige regio’s alleen voor de geboorteco-
horten 1883-1922 (n = 8296). Tot 1940 bestond het bevolkingsregister uit vastbladige registers en gezins-
kaarten, daarna uit persoonskaarten. Met de gezinskaarten is het mogelijk om
op ieder gewenst moment vast te stellen met welke andere personen een onder-
zoekspersoon in een huishouden verblijft. Op de persoonskaart staat geen infor-
matie vermeld over alle personen die met de onderzoekspersoon in een huis-
houden hebben verbleven. Om die reden is besloten om slechts informatie die
in de vastbladige registers en op de gezinskaarten staat vermeld (d.w.z. tot 1940)
te gebruiken en geen poging te ondernemen om de leefvormtrajecten ook na
die datum te construeren. Dit leidt ertoe dat wij ons bij het construeren van de
leefvormtrajecten tussen leeftijd 15 en 40 beperken tot personen geboren tussen
1850 en 1899 (n = 4651). Op basis van deze informatie zijn negen leefvormen onderscheiden waarin
men zich tussen het vijftiende en het veertigste levensjaar kan bevinden. Deze
staan vermeld in tabel 1. Allereerst kan men alleen wonen (A). In de tweede plaats kan men bij de
ouders wonen, al dan niet met andere familieleden, zoals broers en zussen, en Tabel 1 Leefvormen en hun acroniemen
Nummer
Acroniem
Beschrijving
1
A
Alleen
2
O
met Ouders
3
P
met Partner zonder kinderen
4
PK
met Partner met Kinderen
5
K
zonder partner met Kinderen
6
PO
met Partner en (schoon)Ouders
7
PKO
met Partner, Kinderen en (schoon)Ouders
8
F
met Familie anders dan partner, (schoon)ouders of kinderen
9
N
met Niet-familie
10
D
Dood
11
U
Onbekend Tabel 1 Leefvormen en hun acroniemen hilde bras, aart c. liefbroer en cees h. elzinga al dan niet met niet-verwanten, zoals inwonend personeel. Deze leefvorm wordt
aangeduid als ‘met Ouders’ (O). Woont men samen met een partner, maar zon-
der dat een ouder of beide ouders ook deel uitmaken van het huishouden, dan
zijn er twee mogelijkheden. Heeft men geen kinderen, dan wordt de leefvorm
geclassificeerd als ‘met Partner, zonder kinderen’ (P). Heeft men wel kinderen,
dan wordt de leefvorm ingedeeld als ‘met Partner en Kinderen’ (PK). In beide
gevallen maakt het niet uit of er – met uitzondering van de ouders – nog andere
verwanten of niet-verwanten deel uitmaken van het huishouden. Woont men
wel samen met een of meer van zijn of haar kinderen, maar woont er geen part-
ner of ouder in hetzelfde huishouden, dan wordt de leefvorm geclassificeerd als
‘zonder partner, met Kinderen’ (K). Wonen er behalve de partner en eventuele
kinderen ook een of beide ouders in hetzelfde huishouden als de onderzoeksper-
soon, dan wordt de leefvorm ofwel geclassificeerd als ‘met Partner en (schoon)
Ouder’ (PO) of als ‘met Partner, Kinderen en (schoon)Ouder’ (PKO). Opnieuw doet
het niet ter zake of ook andere verwanten of niet-verwanten deel uitmaken van
het huishouden. Wanneer men noch alleen, noch met ouders, kinderen of part-
ner in eenzelfde huishouden woont, dan blijven er nog twee mogelijkheden over:
men woont met andere familieleden, zoals een broer of een zus of een tante
en een oom – en eventuele niet-verwanten – in een huishouden, of men woont
alleen met niet-verwanten, bijvoorbeeld als inwonende bediende of knecht of in
een institutioneel huishouden zoals een klooster. De eerste mogelijkheid wordt
geclassificeerd als ‘met Familie anders dan partner, (schoon)ouders of kinderen’
(F), de tweede mogelijkheid als ‘met Niet-familie’ (N). Ten slotte is het mogelijk
dat een onderzoekspersoon voor het veertigste levensjaar overlijdt. In dat geval
wordt een extra positie ‘Dood’ (D) toegevoegd. 25 De HSN is een rijke, maar ook een complexe databron. Het afleiden van de
leefvormtrajecten uit de basisbestanden van de HSN is dan ook niet eenvoudig. Uit zowel statische als veranderende gegevens die in het bevolkingsregister zijn
opgenomen is per maand afgeleid met welke andere personen de onderzoeks-
persoon in hetzelfde huishouden verbleef. Op basis van deze informatie is ver-
volgens maandelijks bepaald in welke leefvorm men zich bevond.1 Bij de constructie van de leefvormtrajecten treedt een aantal complicaties
op. hilde bras, aart c. liefbroer en cees h. elzinga In de eerste plaats is het vaak niet eenvoudig om per maand te bepalen in
welk huishouden men zich bevindt, omdat de inschrijvings- en uitschrijvings-
maanden in de diverse huishoudens elkaar soms overlappen. Zo kan het zijn
dat een onderzoekspersoon in een huishouden is uitgeschreven per maart 1878,
terwijl dezelfde persoon al per december 1877 is ingeschreven in een ander huis-
houden. In zo’n geval is de inschrijvingsdatum in het laatst genoemde huishou-
den tevens als uitschrijvingsdatum uit het eerdergenoemde huishouden geno-
men. Het kan echter ook zijn dat het verblijf in een huishouden geheel binnen
de inschrijvingsduur in een ander huishouden valt. Het komt bijvoorbeeld voor
dat men in één huishouden staat ingeschreven tussen maart 1866 en december
1891, maar tevens tussen februari 1881 en maart 1882 in een ander huishouden. standaardisering van leefvormen? Dit kan bijvoorbeeld een gevolg zijn van het feit dat een kind bij de ouders thuis
woont, vervolgens een periode als knecht of dienstbode elders verblijft zonder
dat de ouders het kind uitschrijven, waarna het kind weer terugkeert in het
ouderlijk huis. In dergelijke gevallen wordt ervan uitgegaan dat het verblijf in
het huishouden met de vroegste inschrijvingsdatum onderbroken is door het
verblijf in het huishouden met de latere inschrijvingsdatum. 26 26 Een tweede complicatie is dat personen die inwonend zijn bij niet-verwan-
ten of in instituties wonen veelal als alleenstaand worden aangemerkt. Dit leidt
ertoe dat het aandeel alleenstaanden onder het totaal van alle leefvormen bij-
zonder hoog ligt. Om hiervoor te corrigeren is nagegaan of schijnbaar alleen-
staande personen wellicht in registers vermeld staan die suggereren dat zij toch
als dienstboden, knechten of als wonend in een institutie met niet-verwanten
hebben gewoond. Ook is het beroep van schijnbaar alleenstaanden geraadpleegd
om na te gaan of dit mogelijk aanwijzingen opleverde dat een persoon een zoda-
nig beroep had dat het zeer waarschijnlijk is dat deze persoon inwonend was. Als
er op basis van bovenstaande acties aanwijzingen zijn om te veronderstellen dat
een schijnbaar alleenstaande waarschijnlijk inwonend zou kunnen zijn, is deze
persoon alsnog als ‘inwonend bij niet-verwanten’ geclassificeerd. Constructie van de overige variabelen g
Bij de bestudering van de variatie in de leefvormtrajecten worden diverse mogelijk
relevante kenmerken meegenomen. Allereerst gaat het om de reeds eerder bespro-
ken kenmerken geboortecohort, sekse, sociale klasse en religie. hilde bras, aart c. liefbroer en cees h. elzinga Daarnaast worden
de regio waar men woonde ten tijde van de geboorte en de stedelijkheid van de
geboorteplaats als controlevariabelen meegenomen. De belangrijkste reden om
deze twee laatste variabelen als controlevariabelen op te nemen is dat de regionale
spreiding van de onderzoekspersonen zeer ongelijk verdeeld is over de cohorten. Alleen voor Zeeland, Friesland, Utrecht en de gemeente Rotterdam zijn gegevens
over alle cohorten in de huidige HSN-database aanwezig. Bij de operationalisering
van elk van deze kenmerken – met uitzondering van sekse – staan wij kort stil. Geboortecohort. Zoals eerder vermeld, zijn de onderzoekspersonen geboren
tussen 1850 en 1899. Een voor de hand liggend inhoudelijk criterium om tot een
nadere indeling in een beperkt aantal geboortecohorten te komen, ontbreekt. Om na te kunnen gaan of veranderingen tussen cohorten gradueel of meer
sprongsgewijs plaatsvinden, is gekozen voor een indeling in vijf elkaar opvolgen-
de tienjaarlijkse cohorten, te weten 1850-1859, 1860-1869, 1870-1879, 1880-1889
en 1890-1899. In de multivariate analyses wordt het geboortecohort 1870-1879 als
referentiecategorie gehanteerd. Beroepsgroep vader. Om discrepanties tussen sociale groepen te onderzoeken
is een variabele geconstrueerd die sociale groep op basis van de beroepsgroep
van de vader van de onderzoekspersoon indiceert. We zijn steeds uitgegaan van
de beroepstitel van de vader zoals die op de geboorteakte van de onderzoeks- hilde bras, aart c. liefbroer en cees h. elzinga persoon is vermeld. Wanneer de vader niet aanwezig was bij de aangifte van de
geboorte (en er dus geen beroep op de geboorteakte stond), is de eerste beroeps-
titel van de vader uit het bevolkingsregister tijdens de eerste vijf levensjaren van
de onderzoekspersoon gebruikt. Beroepstitels zijn gecodeerd in HISCO (Van Leeu-
wen, Maas & Miles, 2002) en vervolgens omgezet in beroepsklassen aan de hand
van de HISCLASS-indeling (Maas & Van Leeuwen, 2004; Van Leeuwen & Maas,
2005). De oorspronkelijke twaalf categorieën worden samengevat tot acht: ‘Hoge-
re managers en vrije beroepen’, ‘Lagere managers en vrije beroepen, klerken en
winkelpersoneel’, ‘Geschoolde arbeiders’, ‘Boeren en vissers’, ‘Laaggeschoolde
arbeiders’, ‘Ongeschoolde arbeiders’, ‘Landarbeiders’, en ‘Beroep onbekend’. De
laatste groep is maar klein, omdat van de meeste vaders een beroep kon wor-
den gevonden op de geboorteakte of in het bevolkingsregister (zie tabel 2). In de
multivariate analyses fungeert de categorie ‘Lagere managers en vrije beroepen,
klerken en winkelpersoneel’ als referentiecategorie. 27 p
g
Religie ouders. Verschillen in leefvormtrajecten naar religieuze herkomst zijn
onderzocht aan de hand van de kerkelijke gezindte van beide ouders. hilde bras, aart c. liefbroer en cees h. elzinga Er is daar-
bij gebruikgemaakt van de aanduidingen afkomstig uit het bevolkingsregister
ten tijde van de geboorte van de onderzoekspersoon. Op basis van beider reli-
gies zijn in navolging van de indeling van Van Bavel en Kok (2005) de volgende
groepen samengesteld (zie ook Knippenberg, 1992). Ten eerste is er een categorie
‘vrijzinnig-protestanten’ geconstrueerd waaronder zowel de meerderheid van
de gematigde Nederlands Hervormde Kerk valt (voornamelijk de ‘ethische’ en
‘vrijzinnige’ richtingen) als relatief liberale protestantse kerkgenootschappen
zoals de mennonieten, de lutheranen en de remonstranten. Onder de categorie
‘katholiek’ vallen onderzoekspersonen van wie beide ouders rooms-katholiek,
oud-katholiek of vrij-katholiek waren. Een derde categorie omvat degenen van
wie ten minste een ouder tot een orthodox-protestantse kerkelijke gezindte
behoorde. De orthodoxen omvatten de verschillende calvinistische kerkgenoot-
schappen die zich afsplitsten van de Nederlands Hervormde Kerk. Daarnaast
zijn alle Nederlands-hervormden uit gemeenten waar een orthodoxe predikant
was aangesteld (de zogenaamde ‘confessionelen’ en ‘gereformeerde bonders’)
als orthodox-protestants gecodeerd om op die manier de orthodoxe richtingen
binnen de Nederlands Hervormde Kerk te onderscheiden. Hiertoe is gebruik-
gemaakt van de volkstelling van 1920, waarin voor alle gemeenten informatie
over de religieuze signatuur van predikanten te vinden is. Omdat gemeenten
hun eigen predikant kozen, kan worden aangenomen dat zij in de negentiende
eeuw al orthodox waren (Van Bavel & Kok, 2005). Ten vierde hebben we perso-
nen onderscheiden van wie een ouder vrijzinnig-protestants en een ouder katho-
liek was. Zij zijn in de categorie ‘gemengd’ geplaatst. Een vijfde categorie wordt
gevormd door diegenen van wie de ouders joods waren. Wanneer we van beide
ouders geen geloof wisten, is de religie van de ouders als ‘onbekend’ gecodeerd. Ten slotte is er een categorie ‘anders’ geconstrueerd waaronder personen vallen
van wie de ouders onkerkelijk waren of tot een progressieve afscheidingsbewe- standaardisering van leefvormen? ging behoorden. De categorie ‘vrijzinnig-protestants’ wordt in de multivariate
analyses als referentiecategorie gebruikt. Regio geboorteplaats. Om regionale verschillen in de standaardisering van
leefvormtrajecten te onderscheiden zijn respondenten in vier regio’s opgedeeld
die in demografisch opzicht (huwelijkssluiting, vruchtbaarheid en sterfte) en
wat betreft patronen van samenwoning van elkaar verschilden. Als leidraad is
de indeling van Hofstee (Hofstee, 1981) gebruikt (zie ook Boonstra en Van der
Woude, 1984). hilde bras, aart c. liefbroer en cees h. elzinga Vier regio’s werden onderscheiden: het ‘westen en zuidwesten’
(Noord-Holland beneden het IJ, Zuid-Holland, westelijk Utrecht, Zeeland), het
‘noordwesten en noorden’ (Noord-Holland boven het IJ, Groningen, Friesland),
de ‘oostelijke zandgronden’ (Drenthe, Overijssel, grootste deel van Gelderland
– van het oosten tot en met de Veluwe) en de ‘zuidelijke zandgronden en rivier-
kleigebied’ (Limburg, Noord-Brabant, zuidelijk Gelderland en oostelijk Utrecht). De onderzoekspersonen zijn op grond van hun geboorteplaats bij een van deze
regio’s ingedeeld. In de multivariate regressieanalyses is het ‘noordwesten en
noorden’ als referentiecategorie gehanteerd. 28 28 Urbanisatiegraad geboorteplaats. Ten slotte is de urbanisatiegraad van de geboor-
teplaats als variabele meegenomen om te controleren voor verschillen tussen per-
sonen afkomstig uit een stedelijke dan wel rurale context. De urbanisatiegraad
is gebaseerd op het bevolkingsaantal van de geboorteplaats van de onderzoeks-
persoon rond zijn of haar geboortejaar en het percentage van de mannelijke
beroepsbevolking dat werkzaam was in de landbouw in de betreffende gemeente
in die periode. Gemeenten met minder dan 5.000 inwoners zijn als ruraal geco-
deerd en gemeenten met meer dan 20.000 inwoners als urbaan. Gemeenten met
een inwonertal van meer dan 5.000 inwoners en met meer dan 40 procent van
de beroepsbevolking werkzaam in de landbouw zijn als ruraal gecategoriseerd,
terwijl gemeenten met een bevolking tussen 5.000 en 20.000 inwoners, maar met
minder dan 40 procent boeren, als urbaan zijn aangemerkt. De categorie ‘ruraal’
fungeert in de multivariate analyses als referentiecategorie. g
y
g
In tabel 2 is beschrijvende informatie over de verdeling van deze variabelen
over de verschillende cohorten gepresenteerd. Methoden In deze paragraaf beschrijven we achtereenvolgens hoe we turbulentie van en
similariteit tussen leefvormsequenties kwantificeren en ten slotte hoe we de
leefvormtrajecten sorteren in klassen of clusters. De sequenties bestaan uit kortere of langere opeenvolgingen van leefvor-
men en aan iedere leefvorm is een tijdsduur gekoppeld. Een typisch voorbeeld
van zo’n sequentie is de reeks O/54 N/58 P/14 PK/174. Deze onderzoekspersoon
heeft sinds het vijftiende levensjaar eerst 54 maanden bij de ouders ingewoond,
daarna 58 maanden bij een werkgever, heeft vervolgens veertien maanden met
een partner samengeleefd om tenslotte met partner en een of meer kinderen
tenminste tot het veertigste levensjaar samen te wonen. hilde bras, aart c. liefbroer en cees h. elzinga Tabel 2 Proporties per categorie van de achtergrondvariabelen sekse, beroepsgroep vader,
kerkelijke gezindte ouders, regio en urbanisatiegraad, per geboortecohort Tabel 2 Proporties per categorie van de achtergrondvariabelen sekse, beroepsgroep vader,
kerkelijke gezindte ouders, regio en urbanisatiegraad, per geboortecohort kerkelijke gezindte ouders, regio en urbanisatiegraad, per geboortecohort
1850-1859 1860-1869 1870-1879 1880-1889 1890-1899
Totaal
Sekse (man)
51
49
50
47
48
48
Beroepsgroep vader
Hogere managers en
vrije beroepen
2
2
3
7
10
7
Lagere managers en
vrije beroepen,
klerken en winkel-
personeel
19
22
24
17
16
18
Geschoolde
arbeiders
20
16
15
18
17
17
Boeren en vissers
14
16
12
15
13
14
Laaggeschoolde
arbeiders
8
9
6
10
10
9
Ongeschoolde
arbeiders
8
8
8
10
12
10
Landarbeiders
27
25
30
23
21
24
Onbekend
2
2
3
1
1
1
Kerkelijke gezindte
ouders
Beiden vrijzinnig-
protestants
63
60
57
48
45
51
Beiden katholiek
17
19
17
26
27
23
Minstens één ortho-
dox-protestants
11
11
15
14
16
14
Gemengd
5
3
3
4
4
4
Beiden joods
1
1
1
2
2
2
Beiden onbekend
1
2
1
3
2
2
Anders
2
3
5
4
4
4
Regio
Westen en
zuidwesten
50
50
49
47
47
48
Noordwesten en
noorden
31
33
35
22
20
25
Oostelijke
zandgronden
-
-
-
9
12
7
Zuidelijke zandgron-
den en rivierklei
19
17
16
22
21
20
Urbanisatiegraad
(urbaan)
40
40
39
46
50
45
N
463
612
647
1241
1688
4651
Bron: HSN dataset Levenslopen, release 2007.01. 29 standaardisering van leefvormen? Soms is het nuttig om over de opeenvolging van leefvormen te praten zonder
daarbij de verschillende tijdsduren te betrekken. Methoden Zo’n opeenvolging van louter
leefvormen zullen we dan een ‘patroon’ noemen; bijvoorbeeld O N P PK. Iedere sequentie in de data beslaat steeds een even lange periode: 300
maanden (de tijdsduur in maanden tussen vijftien en veertig jaar). De sequen-
ties verschillen echter sterk in lengte: sommige sequenties bestaan uit slechts
één leefvorm, andere daarentegen zijn veel langer, doordat er veel meer leef-
vormen in voorkomen. Hoe langer de opeenvolgingen zijn en hoe meer ver-
schillende leefvormen erin voorkomen, hoe afwisselender of ‘turbulenter’ de
sequentie of het patroon. Het patroon O N is minder turbulent dan het patroon
O N O P PK P A P. Afwisseling wordt ook bepaald door de verdeling van de tijd
over de leefvormen: in de (sub-)sequentie O/99 N/1 speelt de leefvorm N slechts
een minimale rol, want ze beslaat slechts 1 % van de totale duur, terwijl N in
O/50 N/50 een veel belangrijker rol speelt. O/50 N/50 is afwisselender, turbulen-
ter dan O/99 N/1. 30 Om nu het begrip ‘turbulentie’ te kunnen kwantificeren hebben we het
begrip ‘subsequentie’ nodig: we kunnen uit een sequentie een subsequentie vor-
men door uit de sequentie een aantal leefvormen te verwijderen. Als voorbeeld
kijken we naar het veelvoorkomende patroon O P PK. De subsequenties van O P
PK zijn O, P, PK, O P, O PK, P PK en O P PK Merk op dat leefvormen die opeenvolgend zijn in een subsequentie, dat niet hoe-
ven te zijn in de oorspronkelijke sequentie. Niet alle sequenties of patronen van
dezelfde lengte hebben evenveel verschillende subsequenties: de lezer ziet zelf
gemakkelijk in dat O N O er zes heeft en dat O N P zeven subsequenties telt. Een maat voor de afwisseling of turbulentie in een patroon is nu het aantal
verschillende subsequenties dat daarin voorkomt. Ook in de discrete wiskunde
worden aantallen subsequenties of substrings (subsequenties waarin de opeen-
volgende symbolen ook in de oorspronkelijke sequentie opeenvolgend zijn)
gebruikt om de ‘complexiteit’ van patronen te kwantificeren (De Luca, 1999;
Iványi, 1987). In trajecten is echter, anders dan in patronen, aan iedere leefvorm
ook een tijdsduur gekoppeld. Elzinga en Liefbroer (2007) stelden dan ook een cor-
rectie voor waarmee bij de bepaling van de turbulentie ook rekening kan wor-
den gehouden met de variatie in de aan de leefvormen gekoppelde tijdsduur. Die
correctie komt erop neer dat het aantal subsequenties vermenigvuldigd wordt
met de inverse van de verhouding tussen de geobserveerde variantie en de maxi-
male variantie van de tijdsduren. De zo resulterende maat voor turbulentie T
van leefvormtrajecten is ook in dit hoofdstuk gebruikt: 0
2
≤
≤
T
n
log ( ) , waarin n
het aantal subsequenties is dat in de sequentie voorkomt. Voor meer details over
turbulentie verwijzen we de lezer naar Elzinga en Liefbroer (2007). Het kwantificeren van afstanden en similariteit tussen patronen is een pro- hilde bras, aart c. liefbroer en cees h. elzinga bleem dat in verschillende disciplines en om substantiële redenen heel verschil-
lend is aangepakt. Klassiek is de Hamming-afstand (Hamming, 1950), een nog
steeds vaak gebruikte metriek om verschillen tussen patronen te kwantificeren. De Hamming-afstand is gelijk aan het aantal plaatsen waar twee patronen onge-
lijk zijn en heeft ook in de sociale wetenschappen toepassing gevonden (Heiser
& Meulman, 1997). In de context van het analyseren van trajecten en carrières
is de Hamming-afstand echter ongeschikt (Elzinga, 2005). Een generalisatie van
de Hamming-afstand is de Levenshtein-afstand (Levenshtein, 1966), in de sociale
wetenschappen geïntroduceerd door Abbott en Forrest (1986) als Optimal Mat-
ching (OM). De Levenshtein-afstand tussen twee patronen is het minimale aantal
(gewogen) veranderingen dat het ene patroon identiek maakt aan het andere. O, P, PK, O P, O PK, P PK en O P PK Recente inleidingen over OM en de toepassing ervan in de sociale wetenschap-
pen zijn de overzichten van bijvoorbeeld Abbott en Tsay (2000) en van Brüderl en
Scherer (2005). Hoewel OM nog steeds een populaire techniek is, is het gebruik
ervan in de sociale wetenschappen recentelijk uitvoerig bekritiseerd, onder
anderen door Wu (2000) en Elzinga (2003). 31 In dit hoofdstuk gebruiken we een techniek om de mate van similariteit tus-
sen levenslopen vast te stellen die recent door Elzinga (2005) werd ontwikkeld
en ook gebruikmaakt van het begrip ‘subsequentie’. De methode is gebaseerd
op het aantal verschillende gemeenschappelijke subsequenties van twee patro-
nen x en y, gewogen voor de frequentie waarmee die subsequenties in ieder van
de patronen voorkomen. Bijvoorbeeld: de gemeenschappelijke subsequenties
van de patronen O N O en O N P zijn ‘’, O, N en O N. Gewogen voor hun fre-
quenties bedraagt het aantal gemeenschappelijke subsequenties echter 5 = 1 +
2 + 1 + 1, omdat de subsequentie O twee keer voorkomt in O N O. Met behulp
van het aantal gewogen gemeenschappelijke subsequenties kunnen een simila-
riteit 0
1
≤
≤
s x y
( , )
en een afstand d x y
s x y
( , )
( , )
=
−
1
tussen de patronen worden
bepaald. Op verschillende manieren kan nu rekening worden gehouden met de
duur van iedere leefvorm in de trajecten; we gaan daar hier niet verder op in en
verwijzen de lezer naar bijvoorbeeld Elzinga en Liefbroer (2007). Details van de
metriek D en de similariteit s zijn te vinden in Elzinga (2003, 2005) en algorit-
men voor het tellen van (gewogen gemeenschappelijke) subsequenties in Elzin-
ga, Rahmann en Wang (2008). Hier is slechts essentieel dat een grotere waarde
van s x y
( , ) wijst op een grotere gelijkenis tussen de sequenties; s x y
( , ) = 1 als de
sequenties identiek zijn en s x y
( , ) = 0 als de sequenties geen enkele gemeen-
schappelijke subsequentie hebben. Toepassing van de boven geschetste methoden voor het kwantificeren van
afstanden leidt ertoe dat er voor een verzameling van n sequenties een n
n
×
-tabel van afstanden tussen de trajecten wordt berekend. Op basis van die tabel
kunnen de trajecten worden gesorteerd in klassen of clusters van min of meer
gelijkende, dat wil zeggen dichter bijeen liggende trajecten. O, P, PK, O P, O PK, P PK en O P PK Hoe, gegeven een
tabel van afstanden, die clusters eruit zullen zien is natuurlijk afhankelijk van
de regel die men hanteert om trajecten bijeen te brengen in eenzelfde cluster of standaardisering van leefvormen? klasse of juist te scheiden in verschillende clusters. Hoe goed dat lukt hangt in
hoge mate af van de structuur van de tabel van afstanden en van het aantal clus-
ters dat men wenst te vormen: hoe meer clusters, hoe homogener die clusters
zullen zijn (Duda, Hart & Stork, 2001; Tibshirani, Walther & Hastie, 2001). klasse of juist te scheiden in verschillende clusters. Hoe goed dat lukt hangt in
hoge mate af van de structuur van de tabel van afstanden en van het aantal clus-
ters dat men wenst te vormen: hoe meer clusters, hoe homogener die clusters
zullen zijn (Duda, Hart & Stork, 2001; Tibshirani, Walther & Hastie, 2001). 32 De door ons gebruikte techniek staat bekend als K-means clustering. Het itera-
tieve K-means-algoritme berekent, gegeven een vooraf ingesteld aantal (K) clusters
en een initiële partitie, een zodanige partitie dat de som van de afstanden tussen
de centroïden (gemiddelden) van de clusters maximaal is. Hoe goed dat lukt, kun-
nen we kwantificeren als de verhouding 0
1
2
≤
≤
R
van de som van de (gekwa-
drateerde) afstanden tussen de centroïden en de som van alle (gekwadrateerde)
afstanden. Als de zo gevonden clusters perfect homogeen zouden zijn, bereikt R2
de maximale waarde van 1. Een potentieel probleem van deze clustertechniek is
dat het K-means-algoritme weliswaar altijd naar een optimum convergeert, maar
dat dat optimum helaas niet noodzakelijk de best mogelijke oplossing van het
probleem hoeft te zijn. Om toch te waarborgen dat men heel dicht in de buurt
van de beste oplossing komt kan men een groot aantal willekeurig gekozen initi-
ële configuraties gebruiken en die oplossing kiezen die de grootste R2 oplevert. Dat is wat wij hebben gedaan door steeds de beste oplossing te nemen uit een
verzameling van honderd oplossingen van initiële configuraties. Resultaten Turbulentie van trajecten naar volwassenheid Onze eerste hypothese stelt dat het niveau van turbulentie in leefvormtrajecten
naar de volwassenheid afnam voor cohorten geboren tussen 1850 en 1900. Hierin
wordt het idee weerspiegeld dat standaardisering samenging met een afname
van de complexiteit van sequenties. Concreet betekent dat een afname van het
aantal transities, en/of van het aantal verschillende leefvormposities en/of van
de variatie in de timing van transities en de duur van posities. Onze maat om
turbulentie te berekenen houdt zowel rekening met de volgorde van leefvormen
als met de duur die men doorbracht in een bepaald woonarrangement (Elzin-
ga & Liefbroer, 2007). We gebruiken deze maat om de gemiddelde turbulentie
van leefvormtrajecten per geboortecohort te berekenen. De resultaten in tabel
3 laten zien dat trajecten naar volwassenheid van het jongste cohort (1890-1899)
inderdaad significant minder complex waren dan die van onderzoekspersonen
geboren tussen 1850 en 1859. De turbulentie neemt echter niet lineair af, maar
met horten en stoten. Er is sprake van een afname tussen de cohorten 1850-1859
en 1860-1869, maar bij het daaropvolgende cohort (1870-1879) ligt de turbulentie
weer hoger. De daaropvolgende twee cohorten laten echter wel een consistente
daling zien. Verwacht werd dat standaardisering van leefvormtrajecten zich bij mannen
het meest pregnant voordeed. Deze veronderstelling wordt echter niet door de hilde bras, aart c. liefbroer en cees h. elzinga resultaten bevestigd; juist bij vrouwen is er sprake van een significante trend
naar een uniformere jongvolwassenheid. Tijdens de tweede helft van de negen-
tiende eeuw bracht nog minstens 40 procent van alle ongehuwde vrouwen een
deel van haar leven als dienstmeisje door. De levensfase van dienen werd geken-
merkt door veel korte periodes – van vaak een jaar – als inwonend personeelslid
bij een werkgever, meestal afgewisseld met periodes dat vrouwen weer woonach-
tig waren in het ouderlijk huis (Bras, 2002). Trajecten naar volwassenheid veran-
derden drastisch toen de beroepsmogelijkheden voor vrouwen vanaf het begin
van de twintigste eeuw toenamen. Behalve werk in fabrieken, waren er onder
andere banen in grootwinkelbedrijven, als verpleegster, onderwijzeres, typiste
en telegrafiste. Kenmerkend voor deze nieuwe vrouwenberoepen was dat ze geen
inwoning meer vereisten (De Regt, 1993b). Door de toegenomen transportmoge-
lijkheden kon men werk op enige afstand ook steeds beter bereizen. Bovendien
verspreidde het burgerlijk ideaal van huisvrouw en moeder zich steeds meer. Niet alleen onder gehuwde vrouwen was de arbeidsparticipatie laag, dit gold
voor alle vrouwen (De Regt, 1993b). Resultaten Wanneer gezinnen het zich konden veroorlo-
ven, werkten ook ongehuwde meisjes niet. 33 We verwachtten dat trajecten naar volwassenheid van jongeren geboren in
arbeidersmilieus het meest standaardiseerden. Bezien we het gemiddelde ver-
loop van de turbulentie per cohort per sociale klasse, dan zien we dat bij jonge-
ren uit laaggeschoolde en ongeschoolde arbeidersmilieus inderdaad sprake was
van standaardisering. Van alle jongeren geboren rond het midden van de negen-
tiende eeuw waren de leefvormtrajecten van jongeren uit laag- en ongeschoolde
arbeidersgezinnen het grilligste. Deze jongeren waren het vaakst genoodzaakt
om voor hun huwelijk te gaan werken om het gezinsinkomen aan te vullen. De
behoefte aan bijverdiensten veranderde niet voor achtereenvolgende cohorten
geboren tijdens de tweede helft van de negentiende eeuw, wel de wijze waarop
jongeren actief waren op de arbeidsmarkt. Een baan vereiste, zoals ook al bij de
vrouwen is betoogd, steeds minder vaak inwoning in het huishouden van een
werkgever. Daarnaast wisten arbeidersjongeren, door de in het kielzog van de
industrialisering toegenomen welvaart, vaker en al op jongere leeftijd economi-
sche zelfstandigheid te bereiken om in het huwelijk te kunnen treden. Door al
deze veranderingen werden hun trajecten naar volwassenheid gemiddeld korter
en homogener. We verwachtten dat standaardisering het sterkst onder jongeren van vrijzin-
nig-protestantse signatuur optrad. Tabel 3 laat zien dat dit niet het geval was. Bij
jongeren afkomstig uit onkerkelijke gezinnen of van wie tenminste een ouder
tot een progressieve afscheidingsbeweging behoorde (categorie ‘anders’), zien
we wel een significante trend naar eenvormigheid. Maar ook onder jongeren
van katholieke afkomst kwam er over de cohorten significant meer regelmaat in
de overgang naar de volwassenheid. Net als bij de vrouwen en de ongeschoolde
arbeiders vond convergentie naar een gemeenschappelijke standaard dus vaak
plaats bij groepen met aanvankelijk zeer grillige trajecten. Zoals we later zul- standaardisering van leefvormen? len zien (in tabellen 6 en 7) is de afgenomen huwelijksleeftijd van katholieke
vrouwen en de daling van de proportie vrijgezellen onder katholieke mannen
waarschijnlijk debet aan het regelmatiger worden van de leefvormtrajecten van
katholieken. y
p
)
n: HSN-dataset Levenslopen, release 2007.01. a Significante veranderingen zijn vetgedrukt. In dat geval overlappen de 90-procent betrouwbaarheids-
intervallen van het eerste cohort (1850-59) en het laatste cohort (1890-99) niet (op basis van one-
way ANOVA posthoc LSD tests). Resultaten 34 34 Tabel 3 Gemiddelde turbulentie van leefvormtrajecten per sekse, beroepsgroep vader en ker-
kelijke gezindte ouders, per geboortecohort el 3 Gemiddelde turbulentie van leefvormtrajecten per sekse, beroepsgroep vader en ker-
kelijke gezindte ouders, per geboortecohort 1850-1859 1860-1869 1870-1879 1880-1889 1890-1899
Totaal
Totaal
5,881a
5,642
5,882
5,674
5,662
5,715
Sekse
Mannen
5,625
5,616
5,956
5,739
5,764
5,751
Vrouwen
6,148
5,666
5,809
5,616
5,568
5,681
Beroepsgroep vader
Hogere managers
en vrije beroepen
5,957
5,724
5,194
5,793
5,754
5,730
Lagere managers
en vrije beroepen,
klerken en winkel-
personeel
5,983
5,603
5,961
5,622
5,716
5,748
Geschoolde arbei-
ders
5,828
5,650
5,998
5,703
5,722
5,754
Boeren en vissers
5,242
5,406
5,814
5,343
5,592
5,484
Laaggeschoolde
arbeiders
6,470
5,916
6,285
5,750
5,427
5,754
Ongeschoolde
arbeiders
6,822
5,490
5,870
5,795
5,694
5,805
Landarbeiders
5,776
5,743
5,637
5,819
5,658
5,721
Onbekend
5,077
5,966
7,759
5,024
5,741
5,999
Kerkelijke gezindte
ouders
Beiden vrijzinnig-
protestants
5,957
5,576
5,920
5,689
5,775
5,768
Beiden katholiek
6,127
5,697
6,041
5,520
5,579
5,663
Minstens één ortho-
dox-protestants
5,270
5,722
5,727
5,651
5,567
5,606
Gemengd
5,330
6,053
5,297
5,694
5,658
5,620
Beiden joods
5,743
6,466
5,468
6,023
6,348
6,111
Beiden onbekend
3,576
5,477
5,750
6,279
4,898
5,560
Anders
6,969
5,697
5,901
5,981
5,378
5,740
N
463
612
647
1241
1688
4651
a Significante veranderingen zijn vetgedrukt. In dat geval overlappen de 90-procent betrouwbaarheids-
intervallen van het eerste cohort (1850-59) en het laatste cohort (1890-99) niet (op basis van one-
ANOVA
th
LSD t
t ) 1850-1859 1860-1869 1870-1879 1880-1889 1890-1899
Totaal a Significante veranderingen zijn vetgedrukt. In dat geval overlappen de 90-procent betrouwbaarheids-
intervallen van het eerste cohort (1850-59) en het laatste cohort (1890-99) niet (op basis van one-
way ANOVA posthoc LSD tests). hilde bras, aart c. liefbroer en cees h. elzinga Similariteit van trajecten naar volwassenheid Similariteit van trajecten naar volwassenheid
Onze tweede hypothese stelt dat trajecten naar volwassenheid meer op elkaar
zijn gaan lijken. Perfecte gelijkenis zou betekenen dat in elk cohort één reeks
van leefvormen volstaat om de trajecten van alle onderzoekspersonen te repre-
senteren. Om de similariteit van trajecten te meten, gebruiken we een similari-
teitsindex die rekening houdt met het aantal verschillende subsequenties, de
frequentie waarin die verschillende subsequenties voorkomen en de duur van
de subsequenties. Als de gemiddelde similariteit dicht bij 1 ligt is de standaar-
disering hoog, als de gemiddelde similariteit dicht bij 0 ligt juist laag. Resultaten In tabel
4 wordt de gemiddelde similariteit van leefvormtrajecten per geboortecohort
gepresenteerd. Alhoewel de gemiddelde similariteit relatief laag is (variërend
tussen 0,1 en 0,4), is er wel degelijk sprake van een trend naar meer gelijkvor-
mige trajecten. Trajecten naar volwassenheid van het jongste cohort (1890-1899)
lijken significant meer op elkaar dan de leefvormpaden van het oudste cohort. 35 Dit geldt ook voor de levenspaden van jongvolwassen mannen en vrouwen
afzonderlijk. Voor beide groepen geldt dat er zich standaardisering heeft voor-
gedaan, in de zin dat leefvormtrajecten meer gelijkenis zijn gaan vertonen. Bij
mannen is de toename van de similariteit tussen het oudste en het jongste cohort
echter het grootst. Dit suggereert dat standaardisering zich sterker voordeed in
de trajecten naar volwassenheid van mannen dan in die van vrouwen. We verwachtten dat trajecten naar volwassenheid van jongeren geboren in
arbeidersmilieus het meest standaardiseerden. Wanneer we de veranderingen in
de similariteit van leefvormtrajecten per sociale klasse bekijken, zien we inder-
daad dat binnen de klassen van laaggeschoolde arbeiders, ongeschoolde arbei-
ders en landarbeiders trajecten naar volwassenheid meer op elkaar gingen lij-
ken. Verder valt op dat ook binnen enkele andere sociale klassen, namelijk bij de
lagere managers en beoefenaars van vrije beroepen, klerken en winkelpersoneel
en bij de boeren en vissers, trajecten meer overeenkomstig werden. Binnen de
laaggeschoolde en ongeschoolde arbeidersklasse is de toename van de similari-
teit relatief echter het grootst. Ons vermoeden was dat leefvormcarrières van vrijzinnig-protestanten in
sterkere mate standaardiseerden dan die van jongeren van andere religieuze
signatuur. Tussen het oudste en het jongste cohort neemt de similariteit van
leefvormtrajecten bij vrijzinnig-protestanten inderdaad significant toe. Dit geldt
echter evenzeer voor jongeren van katholieke huize. Eerder zagen we al dat ook
de turbulentie in de leefvormtrajecten van katholieken significant afnam. Onze
hypothese dat trajecten naar volwassenheid van vrijzinnig-protestanten het meest
convergeerden naar een gemeenschappelijke standaard wordt door de uitkom-
sten over similariteit dus niet bevestigd. standaardisering van leefvormen? n: HSN-dataset Levenslopen, release 2007.01. way ANOVA posthoc LSD tests). intervallen van het eerste cohort (1850-59) en het laatste cohort (1890-99) niet (op basis van on Resultaten Tabel 4 Gemiddelde similariteit van leefvormtrajecten per sekse, beroepsgroep vader en ker-
kelijke gezindte ouders, per geboortecohort
1850-1859 1860-1869 1870-1879 1880-1889 1890-1899
Totaal
Totaal
0,188a
0,209
0,202
0,221
0,242
0,218
Sekse
Mannen
0,198
0,226
0,209
0,243
0,255
0,231
Vrouwen
0,186
0,199
0,204
0,211
0,235
0,211
Beroepsgroep vader
Hogere managers en
vrije beroepen
0,244
0,226
0,327
0,233
0,288
0,252
Lagere managers en
vrije beroepen,
klerken en winkel-
personeel
0,189
0,215
0,221
0,230
0,235
0,218
Geschoolde
arbeiders
0,229
0,202
0,198
0,227
0,247
0,220
Boeren en vissers
0,203
0,262
0,240
0,245
0,253
0,238
Laaggeschoolde
arbeiders
0,160
0,206
0,274
0,249
0,214
0,208
Ongeschoolde
arbeiders
0,219
0,225
0,217
0,240
0,274
0,239
Landarbeiders
0,198
0,221
0,190
0,209
0,247
0,212
Onbekend
0,279
0,212
0,170
0,195
0,232
0,212
Kerkelijke gezindte
ouders
Beiden vrijzinnig-
protestants
0,192
0,229
0,216
0,224
0,244
0,219
Beiden katholiek
0,187
0,228
0,184
0,213
0,238
0,214
Tenminste één
orthodox-protes-
tants
0,226
0,203
0,223
0,217
0,253
0,223
Gemengd
0,259
0,346
0,300
0,292
0,265
0,257
Beiden joods
0,322
0,288
0,479
0,267
0,258
0,229
Beiden onbekend
0,789
0,308
0,210
0,330
0,286
0,256
Anders
0,226
0,266
0,155
0,244
0,265
0,207
N
463
612
647
1241
1688
4651
a Significante veranderingen zijn vetgedrukt. In dat geval overlappen de 90-procent betrouwbaarheids-
intervallen van het eerste cohort (1850-59) en het laatste cohort (1890-99) niet (op basis van one-
way ANOVA posthoc LSD tests). 36 36 hilde bras, aart c. liefbroer en cees h. elzinga Naar een standaardtraject? Tot dusver is standaardisering afgemeten aan de turbulentie en similariteit van
trajecten naar volwassenheid. In deze paragraaf toetsen we onze derde hypothese
die stelt dat de diversiteit in trajecten afneemt en de dominantie van een traject
waarin gezinsvorming centraal staat, toeneemt voor cohorten geboren tussen
1850 en 1900. We doen dit door op basis van onze gegevens een typologie van
leefvormtrajecten te construeren en te onderzoeken in welke mate achtereen-
volgende geboortecohorten die trajecten doorliepen. In een serie van logistische-
regressieanalyses bekijken we ten slotte wat het profiel was van de onderzoeks-
personen die deze trajecten doorliepen. 37 Door middel van clusteranalyse met het K-means-algoritme zijn de onder-
zoekspersonen in min of meer homogene subgroepen verdeeld. Uit de data zijn
acht clusters gegenereerd (R2 = 0,352). Onder elk van de clusters vallen meerdere
sequenties. De clusters hebben elk één meest karakteristieke sequentie, namelijk
het traject dat de kleinste afstand heeft tot de centroïde van het cluster. In tabel
3 presenteren we deze acht clusters. Resultaten De gemiddelde similariteit geeft aan hoe-
zeer de reeksen die tot een bepaald cluster behoren op elkaar lijken. Het cluster ‘vroege stervers’ omvat 8 procent van de steekproef en heeft een
karakteristieke sequentie O/64 D/236, hetgeen een traject aanduidt waarin men
tot het twintigste jaar bij de ouders woonde en daarna overleed. Zo’n 14 procent
van de onderzoekspersonen doorliep een dienstbodetraject. Dit cluster behelst
vrouwen én mannen die voor hun huwelijk een periode inwoonden bij niet-ver-
wanten. Een kenmerkend dienstbodetraject ziet er als volgt uit: O/54 N/58 P/14
PK/174, dat wil zeggen dat een persoon die dit traject doorliep tot ongeveer het
twintigste jaar bij de ouders woonde, daarna circa vijf jaar inwoonde bij werkge-
vers of andere niet-verwanten, vervolgens trouwde, een jaar later het eerste kind
kreeg en vervolgens tot het veertigste jaar samenwoonde met partner en kinde-
ren. Dan zijn er twee trajecten die wij ‘vroege gezinsvormers’ hebben genoemd. Het eerste traject wordt gekenmerkt door de karakteristieke reeks O/104 P/7
PK/189, oftewel: tot op 23 à 24-jarige leeftijd in het ouderlijk huis wonen, daarna
trouwen en een half jaar samen met de partner leven, alvorens kinderen te krij-
gen. Het andere scenario (vroege gezinsvormers/2) komt bijna op hetzelfde neer,
maar men gaat hier meteen vanuit het ouderlijk huis met partner en kinderen
wonen. Waarschijnlijk is dit laatste traject een artefact, veroorzaakt door verschil-
len in de wijze van registratie tussen de bevolkingsregisters en de gezinskaar-
ten. Daarnaast weerspiegelt het waarschijnlijk ook daadwerkelijk een patroon
van inwoning van ouders en hun getrouwde kinderen. Vooral in de grote steden
bestonden tijdens de eerste helft van de twintigste eeuw grote woningtekorten
(Kok, Mandemakers & Wals, 2005). We voegen beide trajecten in de volgende ana-
lyses samen tot één cluster ‘vroege gezinsvormers’ hetgeen in totaal 27 procent
van alle onderzoekspersonen omvat. Een volgend cluster hebben we ‘late gezinsvormers’ genoemd. Onder deze standaardisering van leefvormen? groep vallen vrouwen en mannen die relatief laat – op circa hun 29ste – huwen
en daarna kinderen krijgen. Deze groep late gezinsvormers, in totaal zo’n 12 pro-
cent, is kenmerkend voor het West-Europese huwelijkspatroon. Een kleine groep
van 5 procent trouwt wel, maar relatief laat (met 27 à 28 jaar) en heeft op zijn
of haar veertigste (nog) geen kinderen. We noemen dit cluster de ‘kinderlozen’. Het cluster ‘vrijgezellen’, 9 procent in totaal, bestaat uit personen die op hun
veertigste nog ongehuwd zijn. O=‘met Ouders’; D=’Dood’; N=‘met Niet-familie’; P=‘met Partner zonder kinderen’; PK=‘met Partner met Kinderen’ (zie ook tabel 1). PK=‘met Partner met Kinderen’ (zie ook tabel 1). Resultaten Ook dit traject is typerend voor het West-Europese
huwelijkspatroon van laat en weinig huwen. Ten slotte is er nog een vrij grote
restcategorie die meer dan een kwart van alle onderzoekspersonen omvat en uit
zeer uiteenlopende trajecten bestaat. De gemiddelde similariteit tussen de reek-
sen binnen dit cluster is dan ook zeer laag. 38 38 Een van de dimensies van standaardisering is het afnemen van de diversiteit
van trajecten en de toename van één dominant traject. Uhlenberg observeerde
onder negentiende- en begin twintigste-eeuwse Amerikaanse cohorten een toe-
name van het ‘standaardtraject’ waarin gezinsvorming centraal stond. In hoe-
verre nam onder Nederlandse cohorten de betekenis van het door ons onder-
scheiden vroege gezinsvormingstraject toe als pad naar de volwassenheid? In
figuur 1 zijn per cohort de proporties onderzoekspersonen die de verschillende
trajecten doorliepen afgebeeld. We zien ten eerste dat het vroege gezinsvormingstraject steeds dominanter
wordt. Slechts eenvijfde van het oudste cohort (1850-1859) volgde dit traject, ter-
wijl bijna eenderde van het jongste cohort (1890-1899) vanuit het ouderlijk huis Tabel 5 Trajecten naar volwassenheid gebaseerd op de clusteranalyse j
g
p
y
Type traject
N
Per
cen
tage
Gemid
delde
similariteit
Standaard
deviatie
similariteit
Karakteristieke sequentie a
Vroege stervers
356
7,65
0,664
0,134
O/64
D/236
Dienstboden
657
14,13
0,418
0,099
O/54
N/58
P/14
PK/174
Vroege gezins
vormers /1
718
15,44
0,769
0,111
O/104 P/7
PK/189
Vroege gezins
vormers /2
519
11,16
0,653
0,126
O/108 PK/192
Late gezinsvormers
550
11,83
0,619
0,129
O/167 P/13
PK/120
Kinderlozen
236
5,07
0,560
0,130
O/152 P/148
Vrijgezellen
402
8,64
0,807
0,141
O/300
Restcategorie
1213
26,08
0,083
0,031
O/52
N/12
O/147
PK/89
a O ‘
O d
’ D ’D
d’ N ‘
Ni
f
ili ’ P ‘
P
d
ki d
’ PK=‘met Partner met Kinderen’ (zie ook tabel 1). hilde bras, aart c. liefbroer en cees h. elzinga trouwde en vóór het 25ste jaar partner en kinderen had. Het vroege gezinsvor-
mingstraject neemt vooral vanaf het geboortecohort 1870-1879 sterk in populari-
teit toe. Kennelijk kwam het proces van convergentie naar een ‘moderne’ levens-
loop rond de eeuwwisseling in een stroomversnelling. 39 De bijna parallelle afname van het resttraject is ook een teken van standaar-
disering. Dit cluster omvat zeer uiteenlopende trajecten die bij geen van de ande-
re clusters passen en ook geen duidelijk eigen profiel hebben. De afname van
deze diffuse diversiteit geeft op zichzelf dus al aan dat uniformering plaatsvond. Resultaten Ook hier zien we dat die afname vooral vanaf het cohort 1870-1879 scherper
wordt. Standaardisering is ook zichtbaar in de afname van het aandeel personen
dat op jonge leeftijd stierf. De toename van het aandeel dat tot de ‘kinderlozen’
behoort – personen die op hun veertigste wel een partner maar geen kinderen
hadden – kan zowel duiden op een stijging van de subfecunditeit als gevolg van
slechte gezondheidszorg en fysieke omstandigheden, als op verbreiding van
moderne opvattingen rond geboortebeperking en vruchtbaarheid, zoals in de
bijdrage van Van Bavel, Kok en Engelen in dit boek aan de orde komt. We zullen
straks zien welke groepen het vaakst dit traject volgden. In een gestandaardiseerde levensloop past ook een afname van het aandeel
late gezinsvormers en vrijgezellen. Figuur 1 laat zien dat er nauwelijks sprake
was van een daling van de proportie vrijgezellen; hun aandeel schommelt tus-
sen 8 en 10 procent. Ook wat betreft de groep late gezinsvormers is het beeld
diffuus. We observeren een U-vormig patroon: laat trouwen neemt eerst af als
optie en vanaf het geboortecohort 1870-1879 weer toe. Ook het dienstbodetraject
neemt niet eenvormig af, maar volgt een omgekeerd U-vormig patroon. Geheel
volgens verwachting stijgt het aandeel mannen en vrouwen dat dit traject door-
loopt aanvankelijk. Bij het geboortecohort 1870-1879 bereikt dit traject een piek;
van deze groep volgt ruim 17 procent het dienstbodetraject. In daaropvolgende
cohorten daalt het weer. Duidelijk blijkt dat de toename van het vroege gezinsvormingstraject vooral
ten koste ging van de zeer heterogene trajecten in het restcluster. Alhoewel het
vroege gezinsvormingstraject voor het jongste cohort weliswaar het belangrijk-
ste pad naar de volwassenheid was geworden, bleven late gezinsvorming, een
vrijgezellenbestaan of een fase als inwonend personeelslid, reële opties om de
jongvolwassenheid te overbruggen. Opvallend is dat deze ‘oude’ paden om de
volwassenheid te bereiken niet aan belang inboetten tijdens de periode 1850-
1940. Determinanten van leefvormtrajecten Determinanten van leefvormtrajecten De tot nog toe gepresenteerde resultaten laten zien dat er tijdens de periode
1850-1940 verscheidene wegen naar volwassenheid leidden. De vraag is daarom
gewettigd wat de determinanten waren van die verschillende trajecten. Welke
groepen volgden het vroege gezinsvormingstraject? En wat was het profiel van standaardisering van leefvormen? Figuur 1 Proporties onderzoekspersonen behorende tot elk van de trajecten, per geboorteco-
hort (N = 4651)
Bron: HSN-dataset Levenslopen, release 2007.01. 0
5
10
15
20
25
30
35
1850-1859
1860-1869
1870-1879
1880-1889
1890-1899
Geboortecohort
Restcategorie
Vroege gezinsvormers
Dienstboden
Late gezinsvormers
Vrijgezellen
Vroege stervers
Kinderlozen
% Figuur 1 Proporties onderzoekspersonen behorende tot elk van de trajecten, per geboorteco
hort (N = 4651) Figuur 1 Proporties onderzoekspersonen behorende tot elk van de trajecten, per geboorteco-
hort (N = 4651) 40 40 Geboortecohort Bron: HSN-dataset Levenslopen, release 2007.01. de personen die de andere leefvormtrajecten doorliepen? Om hier meer inzicht
in te krijgen is een serie binomiale logistische-regressieanalyses uitgevoerd,
waarbij steeds een van de trajecten wordt vergeleken met alle anderen. Hierdoor
komt goed tot uiting onder welke categorieën een traject meer dan gemiddeld,
dan wel minder dan gemiddeld voorkwam.2 de personen die de andere leefvormtrajecten doorliepen? Om hier meer inzicht
in te krijgen is een serie binomiale logistische-regressieanalyses uitgevoerd,
waarbij steeds een van de trajecten wordt vergeleken met alle anderen. Hierdoor
komt goed tot uiting onder welke categorieën een traject meer dan gemiddeld,
dan wel minder dan gemiddeld voorkwam.2 In onze vierde hypothese veronderstelden we dat vooral mannen het domi-
nante traject van vroege gezinsvorming naar de volwassenheid aflegden. Uit
analyses met sekse als onafhankelijke variabele (resultaten niet gerapporteerd in
een tabel) bleek er geen significant verschil te zijn tussen mannen en vrouwen
in de kansen om een vroeg gezinsvormingstraject te doorlopen. De andere trajec-
ten waren wel in sterke mate seksespecifiek; mannen hadden een veel grotere
kans om vroeg te sterven, op latere leeftijd een gezin te vormen of kinderloos te
blijven. Vrouwen daarentegen hadden een grotere kans om een dienstbodetra-
ject te doorlopen. Mede door die seksespecificiteit hebben we besloten om door
middel van het toevoegen van interactietermen te onderzoeken of er wellicht
ook sekseverschillen in de determinanten van de leefvormtrajecten waren. In
het uiteindelijke model zijn alleen de statistisch significante interactie-effecten
opgenomen. Ter wille van de leesbaarheid worden ze in de tabellen 6 en 7 afzon-
derlijk voor vrouwen en mannen gepresenteerd. Determinanten van leefvormtrajecten We bespreken achtereenvolgens
de determinanten van het standaardtraject en daarna van de andere trajecten en
gaan daarbij ook in op eventuele sekseverschillen. Onze hypothese was dat de dominantie van een standaardtraject van gezins-
vorming voor cohorten geboren tussen 1850 en 1900 is toegenomen. In figuur 1 hilde bras, aart c. liefbroer en cees h. elzinga zagen we de toename van dit traject, dat met name snel steeg vanaf het cohort
1870-1879. Uit de multivariate analyse blijkt dat opeenvolgende cohorten man-
nen een steeds grotere kans op een vroeg gezinsvormingstraject hadden. Bij de
vrouwen was die toename niet geheel lineair. Bij zowel mannen als vrouwen
zien we daarentegen de scherpe toename vanaf het cohort 1870-1879 terug. Het
jongste cohort 1890-1899 doorliep aanzienlijk vaker het pad van vroege gezins-
vorming dan het cohort 1870-1879. 41 We veronderstelden verder dat jongeren afkomstig uit gezinnen van arbei-
ders vaker via een ‘standaardtraject’ van jong trouwen en kinderen krijgen de
volwassenheid bereikten. Dat was inderdaad het geval. In de tabellen 6 en 7 kan
afgelezen worden dat jongeren uit ongeschoolde arbeidersmilieus aanmerkelijk
meer kans hadden op een traject van vroege gezinsvorming dan de referentie-
categorie van jongeren uit gezinnen van lagere managers, beoefenaars van vrije
beroepen, klerken en winkelpersoneel, en dat jongeren uit landarbeidersgezin-
nen eveneens veel meer kans op een dergelijk traject hadden. Ook boerendoch-
ters doorliepen echter vaker dan gemiddeld een vroeg gezinsvormingstraject. Voor boerenjongens gold dit niet (in de tabellen is dit significante verschil tus-
sen boerendochters en boerenzonen zichtbaar gemaakt door middel van vetge-
drukte coëfficiënten). Onze veronderstelling dat jongeren afkomstig uit gezinnen van vrijzinnig-
protestanten voorlopers waren in de standaardisering van de jongvolwassenheid
wordt ook gedeeltelijk bevestigd. Katholieke jongens en meisjes volgden inder-
daad veel minder vaak dan jeugdigen van vrijzinnig-protestantse signatuur een
traject van vroege gezinsvorming. Evenwel hadden jongelui geboren in gemengd-
religieuze gezinnen een veel grotere kans om jong te trouwen en kinderen te
krijgen dan de vrijzinnig-protestanten. Verder valt op dat het vroege gezinsvor-
mingstraject relatief vaak voorkwam bij personen die in een stad of in het wes-
ten en zuidwesten geboren waren. De trajecten van late gezinsvormers en vrijgezellen behoorden tot het ‘oude’
West-Europese huwelijkspatroon. Het late gezinsvormingstraject kwam veel voor
onder jongeren afkomstig uit gezinnen uit de bourgeoisie en van geschoolde
arbeiders, boeren en vissers. Ook katholieke mannen hadden een grotere kans
om pas op oudere leeftijd te trouwen. Determinanten van leefvormtrajecten Niet huwen kon het gevolg zijn van laat
huwen. Een recente studie heeft echter laten zien dat aan beide patronen gedeel-
telijk andere factoren ten grondslag lagen (Engelen & Kok, 2003). Dat is ook af te
lezen aan onze resultaten. Boerendochters huwden vaker op latere leeftijd. Uit-
stel betekende voor hen echter nog geen afstel. Uiteindelijk eindigde slechts een
klein deel van hen als oude vrijster. Ook dochters van ongeschoolde arbeiders en
van landarbeiders bleven niet vaak ongehuwd. Mannen van boerenkomaf daar-
entegen hadden een verhoogde kans om vrijgezel te blijven. Ook religieuze her-
komst werkte gedeeltelijk anders in op de kansen om laat of nooit te trouwen. Het celibaat kwam onder katholieke meisjes significant vaker voor dan onder
vrijzinnig protestantse vrouwen. Bij mannen was dit verschil tussen de kerke- standaardisering van leefvormen? Determinanten van leefvormtrajecten Tabel 6 Resultaten van de binomiale logistische-regressieanalyses van de determinanten van
trajecten naar volwassenheid van vrouwen: odds ratio’s g
g
y
trajecten naar volwassenheid van vrouwen: odds ratio’s
Vroege
gezins
vomers
Vroege
stervers
Dienst
boden
Late ge-
zinsvor
mers
Kinder
lozen
Vrijge
zellen
Geboortecohort
1850-1859
1,29 a
1,58 *
0,71 ~
1,68 **
0,59 ~
0,90
1860-1869
0,92
1,22
0,99
1,21
0,66
1,05
1870-1879 (ref.)
1,00
1,00
1,00
1,00
1,00
1,00
1880-1889
1,32 *
1,12
0,78 ~
1,07
0,79
0,77
1890-1899
1,34 *
0,94
0,61***
1,19
1,10
0,85
Beroepsgroep vader
Hogere managers en vrije
beroepen
1,11
0,63
1,04
0,93
0,86
1,13
Lagere managers, vrije
beroepen, klerken en
winkelpersoneel (ref.)
1,00
1,00
1,00
1,00
1,00
1,00
Geschoolde arbeiders
1,15
0,58 ~
1,08
1,05
0,85
0,85
Boeren en vissers
1,58 **
0,82
0,66 *
1,40 ~
0,74
0,61 ~
Laaggeschoolde arbeiders 1,02
1,33
0,88
1,04
0,97
0,81
Ongeschoolde arbeiders
1,46 **
0,82
1,73**
0,81
0,59 ~
0,38 **
Landarbeiders
1,34 *
0,86
1,46 *
0,66 *
0,91
0,37 ***
Onbekend
1,18
1,40
1,88 ~
0,57
0,61
0,37
Kerkelijke gezindte ouders
Beiden vrijzinnig-protes-
tants (ref.)
1,00
1,00
1,00
1,00
1,00
1,00
Beiden katholiek
0,68 ***
1,25
1,18
0,97
0,76
1,73 **
Tenminste één orthodox-
protestants
0,94
1,39 *
0,79
1,01
0,94
1,23
Gemengd
1,43 *
0,86
1,06
0,70
1,29
0,41 *
Beiden joods
1,21
0,45
0,52
0,75
1,33
1,71
Onbekend
1,34
0,48
1,13
1,13
0,72
0,65
Anders
0,83
0,80
1,20
1,29
1,30
0,80
Regio
Westen en zuidwesten
1,44***
0,70 *
0,65 **
0,99
1,35
1,22
Noordwesten en noor-
den (ref.)
1,00
1,00
1,00
1,00
1,00
1,00
Oostelijke zandgronden
1,24
0,97
1,22
1,00
0,53
1,05
Zuidelijke zandgronden
en rivierklei
1,08
0,76
1,03
1,32
0,98
1,24 42 42 hilde bras, aart c. liefbroer en cees h. elzinga Vervolg tabel 6 Vroege
gezins
vomers
Vroege
stervers
Dienst
boden
Late ge-
zinsvor
mers
Kinder
lozen
Vrijge
zellen
Urbaan
1,30 **
0,88
0,66 ***
1,06
1,50 *
0,79 ~
Constante
0,28 ***
0,12 ***
0,59 **
0,11 ***
0,06 ***
0,19 ***
N (mannen en vrouwen)
1230
356
657
546
235
408
Nagelkerke R2 (mannen en
vrouwen)
0,05
0,03
0,12
0,05
0,04
0,04
~ = p<0,10; * = p<0,05; ** = p<0,01; *** = p<0,001. a Niet-vetgedrukte coëfficiënten zijn voor mannen en vrouwen hetzelfde. Vetgedrukte coëfficiënten
verschillen significant tussen mannen en vrouwen. Bron: HSN-dataset Levenslopen, release 2007.01. a Niet-vetgedrukte coëfficiënten zijn voor mannen en vrouwen hetzelfde. Determinanten van leefvormtrajecten Vetgedrukte coëfficiënten
verschillen significant tussen mannen en vrouwen. Bron: HSN-dataset Levenslopen, release 2007.01. lijke gezindten niet aanwezig. Opvallend is verder dat jongeren uit gemengd
religieuze gezinnen een kleinere kans hadden om als vrijgezel of oude vrijster
door het leven te gaan. Ten slotte was het celibaat in de periode 1850-1940 vooral
een plattelandsfenomeen, zoals ook in eerder onderzoek is gevonden (Engelen &
Kok, 2003). Een levensfase van inwonen bij een werkgever was een patroon dat vaak
voor kwam tijdens de jeugd van jongeren uit landarbeidersgezinnen; zij werden
veelal knecht of meid bij een boer. Meisjes uit ongeschoolde arbeidersmilieus
hadden evenzeer een grote kans om als personeelslid in te wonen, wellicht als
dienstbode bij de burgerij. Voor jongens uit de ongeschoolde arbeidersklasse was
dit niet het geval; zij beleefden minder vaak een inwonende fase tijdens hun
jongvolwassenheid. Ook boerendochters en -zoons verbleven veel minder fre-
quent als intern personeelslid in een ander huishouden. Jongeren geboren in het
westen en zuidwesten gingen ook minder vaak inwonen. Dit gold ook voor jon-
gens die geboren werden op de oostelijke zandgronden, een gebied waar familie-
bedrijven van kleine boeren domineerden en nauwelijks gebruik werd gemaakt
van ingehuurde arbeidskrachten. Ten slotte was inwonen een verschijnsel dat
vooral voorkwam in de levenslopen van jongeren die op het platteland geboren
werden. Voortijdige sterfte kwam vooral voor onder het oudste cohort. Opvallend is
dat jongeren van orthodox-protestantse huize relatief vaker op jonge leeftijd
stierven. Meisjes uit de geschoolde arbeidersklasse hadden juist een kleinere
kans om op jonge leeftijd te sterven; bij jongens was dit klassenverschil er niet. Ook zijn er regionale verschillen in de determinanten van het vroege sterverstra-
ject. Jongvolwassenen geboren in het zuidwesten en het westen stierven minder
vaak op jonge leeftijd. De kinderlozen, die op hun veertigste wel een partner maar geen kinderen
hadden, volgden wellicht de meest ‘moderne’ levensloop. Over de cohorten heen standaardisering van leefvormen? Determinanten van leefvormtrajecten Tabel 7 Resultaten van de binomiale logistische-regressieanalyses van de determinanten van
trajecten naar volwassenheid van mannen: odds ratio’s trajecten naar volwassenheid van mannen: odds ratio’s
Vroege
gezins
vormers
Vroege
ster
vers
Dienst
boden
Late
gezins
vormers
Kin
derlo
zen
Vrijge
zellen
Geboortecohort
1850-1859
0,62 *a
1,58 *
0,71 ~
1,68 **
0,59 ~
0,90
1860-1869
0,92
1,22
0,99
1,21
0,66
1,05
1870-1879 (ref.)
1,00
1,00
1,00
1,00
1,00
1,00
1880-1889
1,32 *
1,12
0,78 ~
1,07
0,79
0,77
1890-1899
1,34 *
0,94
0,61 ***
1,19
1,10
0,85
Beroepsgroep vader
Hogere managers en vrije
beroepen
1,11
0,63
1,04
0,93
0,86
1,13
Lagere managers en vrije beroepen,
klerken en winkelpersoneel (ref.)
1,00
1,00
1,00
1,00
1,00
1,00
Geschoolde arbeiders
1,15
1,05
1,08
1,05
0,85
0,85
Boeren en vissers
0,84
0,82
0,66 **
1,40 ~
0,74
1,83 **
Laaggeschoolde arbeiders
1,02
1,33
0,88
1,04
0,97
0,81
Ongeschoolde arbeiders
1,46 *
0,82
0,67 *
0,81
0,59 ~
0,93
Landarbeiders
1,34 *
0,86
1,46 *
0,66 *
0,91
0,91
Onbekend
1,18
1,40
1,88 ~
0,57
0,61
0,37
Kerkelijke gezindte ouders
Beiden vrijzinnig-protestants (ref.)
1,00
1,00
1,00
1,00
1,00
1,00
Beiden katholiek
0,68 ***
1,25
1,18
1,43 *
0,76
1,03
Tenminste één orthodox-protes-
tants
0,94
1,39 *
0,79
1,01
0,94
1,23
Gemengd
1,43
0,86
1,06
0,70
1,29
0,41 *
Beiden joods
1,21
0,45
0,52
0,75
1,33
1,71
Onbekend
1,34
0,48
1,13
1,13
0,72
0,65
Anders
0,83
0,80
1,20
1,29
1,30
0,80
Regio
Westen en zuidwesten
1,44 ***
0,70 *
0,42 *** 0,99
1,35
1,22
Noordwesten en noorden (ref.)
1,00
1,00
1,00
1,00
1,00
1,00
Oostelijke zandgronden
1,24
0,97
0,54 ~
1,00
0,53
1,05
Zuidelijke zandgronden en rivierklei
1,08
0,76
1,03
0,73
0,98
1,24
Urbaan
1,30 **
0,88
0,66 ***
1,06
1,50 *
0,79 ~
Constante
0,34
0,12
0,29
0,22
0,08 *** 0,14 ***
N (mannen en vrouwen)
1230
356
657
546
235
408
Nagelkerke R2 (mannen en vrouwen)
0,05
0,03
0,12
0,05
0,04
0,04
~ = p<0,10; * = p<0,05; ** = p<0,01; *** = p<0,001. a
Niet-vetgedrukte coëfficiënten zijn voor mannen en vrouwen hetzelfde. Vetgedrukte coëfficiënten
verschillen significant tussen mannen en vrouwen 44 a
Niet-vetgedrukte coëfficiënten zijn voor mannen en vrouwen hetzelfde. Vetgedrukte coëfficiënten
verschillen significant tussen mannen en vrouwen. Bron: HSN-dataset Levenslopen, release 2007.01. Bron: HSN-dataset Levenslopen, release 2007.01. hilde bras, aart c. liefbroer en cees h. elzinga hilde bras, aart c. liefbroer en cees h. Determinanten van leefvormtrajecten elzinga manifesteerde dit traject zich steeds duidelijker; onder het cohort 1850-1859
kwam het maar half zo vaak voor als bij het cohort 1870-1879. Het was vooral een
traject dat door de middenklasse werd gevolgd en het kwam veel minder vaak
voor onder ongeschoolde arbeiders. Net als het vroege gezinsvormingstraject
manifesteerde kinderloosheid zich vooral onder stedelingen. 45 Conclusie en discussie De vraag of en in hoeverre er sprake was van standaardisering van trajecten naar
volwassenheid tijdens de periode 1850-1940 liep als een rode draad door deze
bijdrage. Aan de hand van onze analyse van de HSN-gegevens kunnen we op deze
vraag nu een antwoord geven. We verwachtten dat standaardisering in de eerste plaats een vermindering
van de complexiteit van leefvormtrajecten betekende. Die complexiteit nam tij-
dens de periode 1850-1940 inderdaad af, zij het met horten en stoten. Pas bij
cohorten geboren vanaf 1880 werden leefvormtrajecten steeds regelmatiger. De
complexiteit in de leefvormtrajecten van vrouwen, van ongeschoolde en laag-
geschoolde arbeiders en van katholieken, die aanvankelijk zeer hoog was, nam
over de cohorten heen het sterkst af. Ook onze tweede hypothese, die stelde dat
trajecten naar volwassenheid meer op elkaar gingen lijken, werd bevestigd. Ook
hier zagen we dat de leefvormtrajecten van met name verschillende categorieën
arbeidersjongeren meer op elkaar gingen lijken. Het feit dat deze groepen toe-
groeiden naar een gemeenschappelijk niveau duidt op standaardisering. Een derde aspect van standaardisering zou de afname van het aantal ver-
schillende soorten trajecten en de toegenomen dominantie van een specifiek
traject zijn geweest. Door middel van clusteranalyse identificeerden we zes tra-
jecten die cohorten geboren tijdens de tweede helft van de negentiende eeuw
vaak volgden: vroeg sterven (tussen het vijftiende en veertigste jaar), inwonen
als knecht of dienstbode, vroege gezinsvorming, late gezinsvorming, een vrijge-
zellenbestaan en kinderloos blijven. Voor achtereenvolgende cohorten werd het
vroege gezinsvormingstraject steeds dominanter. Dit gebeurde echter zonder de
andere patronen geheel weg te drukken; de toename ging vooral ten koste van
een zeer heterogeen resttraject. Andere paden naar de volwassenheid die gebrui-
kelijk waren in de agrarisch-ambachtelijke samenleving, zoals laat huwen of als
oude vrijster of vrijgezel leven, bleven ook voor de jongste cohorten nog heel
gebruikelijk. Via welk traject men de volwassenheid bereikte was mede afhankelijk van
iemands sekse, sociale klasse en religieuze herkomst. Jongeren uit de arbeiders-
klasse, boerendochters, jongeren afkomstig uit gemengd religieuze gezinnen
en stedelingen volgden het vaakst een vroeg gezinsvormingstraject. Zij lijken
voorlopers te zijn geweest in het volgen van een moderne levensloop. Jongeren
afkomstig uit de middenklasse en katholieke mannen waren meestal late gezins-
vormers. Mannen van boerenkomaf, katholieke meisjes en plattelandsjongeren standaardisering van leefvormen? bleven het vaakst vrijgezel. Vrouwen, en vooral die uit ongeschoolde arbeidersmi-
lieus, overbrugden de periode tussen jeugd en huwelijk meestal met een fase als
dienstbode. Noten 1. De gebruikte SPSS-syntax is opvraagbaar bij de auteurs. 2. Een alternatieve procedure is om een multinomiale logistische regressieanalyse uit te voe-
ren, waarbij alle trajecten gelijktijdig onderling vergeleken worden. Dit heeft als voordeel
dat steeds per twee typen trajecten gekeken kan worden of de kans dat men meer of min-
der dan gemiddeld in een van de twee trajecten voorkomt varieert per sociale categorie. Een nadeel is echter dat het aantal vergelijkingen met zeven trajecten erg groot wordt (te
weten 21 paarsgewijze vergelijkingen). Aangezien wij vooral geïnteresseerd zijn in datgene
wat ieder van de trajecten onderscheidde van alle anderen is hier voor een serie binomiale
logistische regressies gekozen. Conclusie en discussie Met name jongeren van orthodox-protestantse huize hadden grotere
kansen om vroeg te sterven. Stedelingen ten slotte bleven het vaakst kinderloos. 46 Onze bijdrage heeft laten zien dat de term ‘standaardlevensloop’ zoals die
door levensloopsociologen vaak wordt gehanteerd om levenslooppatronen in het
verleden mee aan te duiden (nog) niet opgaat voor Nederlanders die geboren wer-
den tijdens de tweede helft van de negentiende eeuw. Zeker, er vond een trend
van uniformering van de fase van jongvolwassenheid plaats; ze werd homogener
en ging voor opeenvolgende generaties meer op elkaar lijken. Maar van één dui-
delijk standaardtraject naar de volwassenheid was nog geen sprake. j
j
g g
p
Het proces van standaardisering lijkt daarmee bestaan te hebben uit twee
fasen. Onder invloed van industrialisering, urbanisatie, en de groei van infra-
structuur tijdens de periode 1850-1940 voltrok zich een eerste ronde van unifor-
mering van de jongvolwassenheid. Standaardisering uitte zich door het verdwij-
nen van traditioneel grillige patronen van inwonen bij niet-familie, terwijl ook
algemenere en eerdere gezinsvorming binnen het bereik van bredere lagen van
de Nederlandse bevolking kwam. Pas na de Tweede Wereldoorlog ging het groei-
ende overheidsapparaat en de wetgeving rond leerplicht, scholing en pensioen
de transitie naar volwassenheid reguleren en vond een verdere standaardisering
plaats. Literatuur Abbott, A. & J. Forrest (1986). Optimal matching methods for historical sequen-
ces. Journal of Interdisciplinary History, 16, 471-494. Abbott, A. & A. Tsay (2000). Sequence analysis and optimal matching methods in
sociology. Sociological Methods & Research, 29, 3-33. Abbott, A. & A. Tsay (2000). Sequence analysis and optimal matching methods in
sociology. Sociological Methods & Research, 29, 3-33. Abbott, A. & A. Tsay (2000). Sequence analysis and optimal matching methods in
sociology. Sociological Methods & Research, 29, 3-33. Baars, J. (1991). Het regime van de klok. Over de chronologisering van de moder-
ne levensloop. Amsterdams Sociologisch Tijdschrift, 18, 100-120. Baars, J. (1991). Het regime van de klok. Over de chronologisering van de moder-
ne levensloop. Amsterdams Sociologisch Tijdschrift, 18, 100-120. Baars, J. (1991). Het regime van de klok. Over de chronologisering van de moder-
ne levensloop. Amsterdams Sociologisch Tijdschrift, 18, 100-120. Blikman-Ruiterkamp, B. (2000). ‘Aachtingswaardige vriendin’. Drieënvijftig brie-
ven van een dienstbode uit Tholen. Archief van het Koninklijk Zeeuwsch Genoot-
schap der Wetenschappen, 25-96. Blikman-Ruiterkamp, B. (2000). ‘Aachtingswaardige vriendin’. Drieënvijftig brie-
ven van een dienstbode uit Tholen. Archief van het Koninklijk Zeeuwsch Genoot-
schap der Wetenschappen, 25-96. Blikman-Ruiterkamp, B. (2000). ‘Aachtingswaardige vriendin’. Drieënvijftig brie-
ven van een dienstbode uit Tholen. Archief van het Koninklijk Zeeuwsch Genoot-
schap der Wetenschappen, 25-96. hilde bras, aart c. liefbroer en cees h. elzinga hilde bras, aart c. liefbroer en cees h. elzinga Boonstra, O.W.A. & A.M. van der Woude (1984). Demographic transition in the
Netherlands. A statistical analysis of regional differences in the level and
development of the birth rate and of fertility, 1850-1890. A.A.G. Bijdragen, 24,
1-57. 47 Bras, H. (2002). Zeeuwse meiden. Dienen in de levensloop van vrouwen. Amsterdam:
Aksant Academic Press. Brinkgreve, C. & A. de Regt (1991). Adolescentie als opgave. Ontwikkelingen in
een levensfase 1750-1990. In I. van der Zanden (red.), Het is meisjes menens. Inlei-
ding meisjesstudies (pp. 15-35). Amersfoort/Leuven: Acco. Brückner, J. & K.U. Mayer (2005). De-standardization of the life course: What
might it mean? And if it means anything, whether it actually took place. In
R. Macmillan (red.), The structure of the life course: Standardized? Individualized? Differentiated? (Vol. 9) (pp. 27-54). Amsterdam: Elsevier. Brüderl, J. & S. Scherer (2005). Methoden zur Analyse von Sequenzdaten. Kölner
Zeitschrift für Soziologie und Sozialpsychologie, 44, 330-347. De Luca, A. (1999). On the combinatorics of finite words. Theoretical Computer Sci-
ence, 218, 13-39. Diederiks, H.A. (red.) (1992). Van agrarische samenleving naar verzorgingsstaat. De
modernisering van West-Europa sinds de vijftiende eeuw. Groningen: Wolters-
Noordhoff. Duda, R.O., P.R. Hart. & D.G. Stork (2001). Pattern classification. New York: Wiley Eijk, D. van (2000). Eigen levens. Standaard-levensloop is een historische uitzon-
dering. NRC Handelsblad, 26 augustus 2000. Elzinga, C.H. (2003). Sequence similarity – A non-aligning technique. Sociological
Methods & Research, 31, 3-29. Elzinga, C.H. (2005). Combinatorial representation of token sequences. Journal of
Classification, 21, 87-118. Elzinga, C.H. & A.C. Liefbroer (2007). De-standardization of family-life trajectories
of young adults: A cross-national comparison using sequence analysis. Euro-
pean Journal of Population, 23(3-4), 225-250. Elzinga, C.H., S. Rahmann & H. Wang (2008). Algorithms for subsequence combi-
natorics. Theoretical Computer Science, in druk. Engelen, T. & J. Kok (2003). Permanent celibacy and late marriage in the Nether-
lands, 1890-1960. Population, 58, 67-96. Falkenburg, P. (1905). De huwelijkskansen der vrouwen in Nederland. Haarlem: Boh Gillis, J.R. (1974). Youth and history. Tradition and change in European age relations
1770-present. New York/London: Academic Press. Glick, P.C. (1947). The family cycle. American Sociological Review, 12, 164-174. 1770-present. New York/London: Academic Press. Glick, P.C. (1947). The family cycle. American Sociological Review, 12, 164-174. p
/
Glick, P.C. (1947). The family cycle. American Sociological Review, 12, 164-174. Glick, P.C. & R.J. Parke (1965). New approaches in studying the life cycle of the
family. hilde bras, aart c. liefbroer en cees h. elzinga Demography, 2, 187-202. Hagestad, G.O. (1992). Assigning rights and duties: Age, duration, and gender in
social institutions. In W.R. Heinz (red.), Theoretical advances in life course research. Vol III: Status passages, institutions, and gatekeeping (pp. 261-279). Weinheim:
Deutscher Studien Verlag. standaardisering van leefvormen? Hagestad, G.O. & B.L. Neugarten (1985). Age and the life course. In R.H. Binstock &
E. Shanas (red.), Handbook of aging and the social sciences (Vol. 2) (pp. 36-61). New
York: Van Nostrand & Reinhold. 48 Hajnal, J. (1965). European marriage patterns in perspective. In D.V. Glass (red.),
Population in history (pp. 101-143). London: Arnold. Hamming, R.W. (1950). Error-detecting and error-correcting codes. Bell System
Technical Journal, 26, 147-160. Hanawalt, B.A. (1992). Historical descriptions and prescriptions for adolescence. Journal of Family History, 17, 341-351. Heiser, W.J. & J.J. Meulman (1997). Representation of binary multivariate data
by graph models using the Hamming metric. In E. Wegman & S. Azen (red.),
Computing science and statistics (pp. 517-525). Fairfax, VA: Interface Foundation
of North America, Inc. Hofstee, E.W. (1981). Korte demografische geschiedenis van Nederland van 1800 tot
heden. Bussum: Unieboek. Hogan, D.P. (1981). Transitions and social change. The early lives of American men. New
York etc.: Academic Press. Hynes, K. & M. Clarkberg (2005). Women’s employment patterns during early
parenthood: A group-based trajectory analysis. Journal of Marriage and Family,
2005, 222-239. Iványi, A. (1987). On the d-complexity of words. Annales Universitatis Scientiarum
Budapestinensis de Rolando Eötvös nominatae, Sectio Computatorica, 8, 69-90. Knippenberg, H. (1992). De religieuze kaart van Nederland. Omvang en geografische
spreiding van de godsdienstige gezindten vanaf de Reformatie tot heden. Assen/Maas-
tricht: Van Gorcum. Knippenberg, H. & B. de Pater (1988). De eenwording van Nederland. Schaalvergroting
en integratie sinds 1800. Nijmegen: SUN. Kohli, M. (1986). The world we forgot: A historical review of the life course. In V. W. Marshall (red.), Later life: The social psychology of ageing (pp. 271-303). Beverly
Hills, CA: Sage. Kok, J., K. Mandemakers & H. Wals (2005). City nomads. Changing residence as a
coping strategy, Amsterdam, 1890-1940. Social Science History, 29, 15-43. Laslett, P. (1977). Characteristics of the western family considered over time. In
P. Laslett, Family life and illicit love in earlier generations: essays in historical socio-
logy. Cambridge: Cambridge University Press. Leeuwen, L.T. van (1987). Leefvormen en individuele levensloop in sociaal-demo-
grafisch perspectief; een inventarisatie van nieuwe patronen. Sociologisch en
antropologisch jaarboek, 130-147. Leeuwen, M.H.D. van (1994). hilde bras, aart c. liefbroer en cees h. elzinga Logic of charity: Poor relief in preindustrial Europe. Journal of Interdisciplinary History, 24, 89-613. Leeuwen, M.H.D. van & I. Maas (2005). A short note on HISCLASS. http://historyof-
work.iisg.nl/docs/hisclass-brief.doc Leeuwen, M.H.D. van, I. Maas & A. Miles (2002). HISCO. Historical International hilde bras, aart c. liefbroer en cees h. elzinga Standard Classification of Occupations. Leuven: Leuven University Press. Levenshtein, V.I. (1966). Binary codes capable of correcting deletions, insertions
and reversals. Soviet Physics Doklady, 10, 707-710. 49 Liefbroer, A.C. & J. de Jong Gierveld (1993). Veranderingen in de overgang van
jeugd naar volwassenheid. Een vergelijking van cohorten geboren tussen
1903 en 1965. In M. Du Bois-Reymond & J. de Jong Gierveld (red.), Volwassen wor-
den. Generaties toen en nu: transities in de levensloop (pp. 17-35). Houten/Zaventem:
Bohn Stafleu Van Loghum. Liefbroer, A.C. & P.A. Dykstra (2000). Levenslopen in verandering. Een studie naar ont-
wikkelingen in de levenslopen van Nederlanders geboren tussen 1900 en 1970. Den
Haag: SDU uitgevers. Liefbroer, A.C. & C.H. Elzinga (2007). Intergenerational transmission of behavi-
oral patterns: Similarity of parent’s and children’s family-life trajectories. Intern rapport VU. Maas, I. & M.H.D. van Leeuwen (2004). Recode job from HISCO into HISCLASS. Mak, G. (2005). Alleen met velen. Het verhaal van mijn moeder. Amsterdam: Forum Mandemakers, K. (1996). HBS en Gymnasium. Ontwikkeling, structuur, sociale ach-
tergrond en schoolprestaties, Nederland, circa 1800-1968. Amsterdam: Stichting
beheer IISG. Mandemakers, K. (2000). The Netherlands. Historical Sample of the Netherlands. In P. Kelly Hall, R. McCaa & G. Thorvaldsen (red.), Handbook of international
historical microdata for population research (pp. 149-177). Minneapolis: Minnesota
Population Center. Mayer, K.U. (1986). Structural constraints on the life course. Human Development,
29, 163-170. Modell, J., F.F.J. Furstenberg & T. Hershberg (1976). Social change and transitions
to adulthood in historical perspective. Journal of Family History, 1, 7-32. Modell, J., F.F.J. Furstenberg & D. Strong (1978). The timing of marriage in the
transition to adulthood: Continuity and change, 1860-1975. The American Jour-
nal of Sociology, 84. Supplement: Turning Points: Historical and Sociological Essays on
the Family, S120-S150. Mouw, T. (2005). Sequences of early adult transitions: A look at variability and
consequences. In R.A.J. Settersten, F.F.J. Furstenberg & R.G. Rumbaut (red.), On
the frontier of adulthood: Theory, research and public policy (pp. 256-291). Chicago:
Chicago University Press. Oort, D. van (2006). Vrouw in de schaduw. Een familiegeschiedenis. Amsterdam: Cos-
see. Pollock, G. (2007). hilde bras, aart c. liefbroer en cees h. elzinga Holistic trajectories: a study of combined employment, hou-
sing and family careers by using multiple-sequence analysis. Journal of the
Royal Statistical Society, 170 (Part 1), 167-183. Poppel, F. van (1974). De differentiële vruchtbaarheid in Nederland in historisch
perspectief: de invloed van sociale status. Bevolking en Gezin, 2, 223-247. Poppel, F. van (1985). Late fertility decline in the Netherlands: the influence of standaardisering van leefvormen? religious denomination, socio-economic group and religion. European Journal
of Population, 1, 347-373. religious denomination, socio-economic group and religion. European Journal
of Population, 1, 347-373. Poppel, F. van (1992a). Religion and health: Catholicism and regional mortality
differences in nineteenth-century Netherlands. Social History of Medicine, 5,
229-253. 50 Poppel, F. van (1992b). Trouwen in Nederland. Een historisch-demografische studie van
de 19e en vroeg-20e eeuw. Den Haag: Stichting NIDI. Poppel, F. van (1999). De ‘statistieke ontleding van de dooden’: een spraakzame bron? Nijmegen: Nijmegen University Press. Regt, A. de (1993a). Arbeiders, burgers en boeren: gezinsleven in de negentiende
eeuw. In T. Zwaan (red.), Familie, huwelijk en gezin in West-Europa (pp. 193-218). Amsterdam/Heerlen: Boom/Open Universiteit. Regt, A. de (1993b). Het ontstaan van het ‘moderne’ gezin, 1900-1950. In T. Zwaan
(red.), Familie, huwelijk en gezin in West-Europa (pp. 219-239). Amsterdam/Heer-
len: Boom/Open Universiteit. Stevens, D.A. (1990). New evidence on the timing of early life course transitions:
The United States 1900 to 1980. Journal of Family History, 15, 163-178. Tibshirani, R., G. Walther & T. Hastie (2001). Estimating the number of data clus-
ters via the Gap statistic. Journal of the Royal Statistical Society, Series B, 63, 411-
423. Uhlenberg, P. (1969). A study of cohort life cycles: Cohorts of native born Massa-
chusets women, 1830-1920. Population Studies, 23, 407-420. Uhlenberg, P. (1974). Cohort variations in family life cycle experiences of U.S. females. Journal of Marriage and the Family, 36, 284-292. Valk, L. van der (1986). Van pauperzorg tot bestaanszekerheid: een onderzoek naar de
ontwikkeling van de armenzorg in Nederland tegen de achtergrond van de overgang
naar de algemene Bijstandswet, 1912-1965. Amsterdam: Stichting IISG Beheer. Van Bavel, J. & J. Kok (2005). The role of religion in the Dutch fertility transition:
starting, spacing and stopping in the heart of the Netherlands, 1845-1945. Continuity and Change, 20, 247-263. Woud, A. van der (2007). Een nieuwe wereld. Het ontstaan van het moderne Nederland. Amsterdam: Bert Bakker. Zanden, J.L. van & A. van Riel (2004). hilde bras, aart c. liefbroer en cees h. elzinga The strictures of inheritance: The Dutch economy
in the nineteenth century. Princeton: Princeton University Press. Wintle, M. (2000). An economic and social history of the Netherlands, 1800-1920. Demo-
graphic, economic and social transition. Cambridge: Cambridge University Press. Wu, L.L. (2000). Some comments on “Sequence analysis and optimal matching
methods in sociology: Review and prospect”. Sociological Methods & Research,
29, 41-64. hilde bras, aart c. liefbroer en cees h. elzinga Hoge kinderloosheid tijdens het interbellum in
Nederland
De rol van godsdienst, levensstandaard en economische crisis Jan Van Bavel, Jan Kok en Theo Engelen Inleiding In de recente literatuur rond de stijging van de kinderloosheid in het Westen
wordt uiterst zelden verwezen naar de stijging die plaatsvond vanaf ongeveer het
laatste kwart van de negentiende eeuw. Nochtans ging de daling van de vrucht-
baarheid ook op het einde van de negentiende en in het begin twintigste eeuw
in de meeste Europese landen en in de Verenigde Staten van Amerika gepaard
met een opmerkelijke toename van de kinderloosheid (Anderson, 1998; Festy,
1979; Rowland, 2007; Dykstra & Hagestad, 2007). De kinderloosheid bereikte haar
voorlopige hoogtepunt tussen de Eerste en de Tweede Wereldoorlog en was toen
voor een niet onbelangrijk deel verantwoordelijk voor het feit dat de vruchtbaar-
heid in vele landen een hele tijd onder het vervangingsniveau lag. Dat was onder
meer het geval in België, Duitsland, Denemarken, Engeland, Frankrijk, Noorwe-
gen, Zweden en Zwitserland, maar niet in Nederland. In Nederland in zijn geheel
dook de vruchtbaarheid toen nog niet onder het vervangingsniveau (Festy, 1979;
Frejka & Sardon, 2004). Dat neemt niet weg dat een aanzienlijk aandeel van de
Nederlandse vrouwen geboren rond het jaar 1900 kinderloos was op 45-jarige
leeftijd, namelijk ongeveer 23 procent volgens de schattingen van Dykstra (net
iets minder dan de Franse 25 procent, maar ruim dubbel zoveel als de 11 procent
kinderloze Nederlandse vrouwen geboren vlak na de Tweede Wereldoorlog; zie
Rowland, 2007, Table 1).1 Mogelijk blijft in de hedendaagse literatuur de vergelijking met de laat
negentiende-eeuwse en vroeg-twintigste-eeuwse evolutie achterwege, omdat
onderzoekers betwijfelen of die historische vergelijking wel relevant is voor de
hedendaagse situatie. De indruk bestaat namelijk dat het om twee categoriek
verschillende vormen van kinderloosheid zou gaan. Vóór ‘de pil’ zou hoge kin-
derloosheid vooral onvrijwillig zijn geweest. Onvruchtbaarheid kon bijvoorbeeld
het gevolg zijn van chronische ziekte of ondervoeding of een te lang uitgesteld
huwelijk (Hakim, 2000; Poston & Trent, 1982). Dat de gemiddelde vruchtbaar- heid tijdens het interbellum zo laag was, had volgens recente interpretaties veel-
eer te maken met de diepe economische en politieke crisis van die jaren (Frejka
& Sardon, 2004, 58; Hakim, 2000; Lesthaeghe & Surkyn, 2004; 2006; Sobotka,
2008). 52 )
Vanaf het midden van de jaren zestig zou dan een heel nieuw type van kin-
derloosheid gegroeid zijn, als structureel onderdeel van de sociaal-demografische
veranderingen bekend als de tweede demografische transitie (Lesthaeghe & Van
de Kaa, 1986). Het internationale debat over lage vruchtbaarheid tijdens het
interbellum Van revolutie naar transitie 53 Na de Tweede Wereldoorlog kwam een heel gestileerde interpretatie van de
grote demografische ontwikkelingen sinds het einde van de achttiende eeuw in
zwang, die van de demografische transitie. In de laatste eeuwen, zo luidt deze
interpretatie, maakte het Westen een overgang mee van een bevolkingseven-
wicht met relatief hoge vruchtbaarheids- en sterftecijfers naar een evenwicht
met lage vruchtbaarheid en sterfte. Omdat de sterftetransitie eerder startte dan
de vruchtbaarheidstransitie, trad er tijdelijk een sterke bevolkingsgroei op. De
vruchtbaarheidstransitie werd dan ook geïnterpreteerd als de logische evolutie
naar een nieuw evenwicht (Coale, 1986). In de jaren zestig dachten vele demogra-
fen en sociologen dat die overgang zo ongeveer was voltooid (Lesthaeghe & Van
de Kaa, 1986). De meeste demografen uit het interbellum interpreteerden de zich voor hun
ogen voltrekkende vruchtbaarheidsdaling niet als een evolutie naar een nieuw
demografisch evenwicht, maar veeleer als een demografische revolutie, om de
term van Landry (1933; 1934) te gebruiken. Het einde van dit proces zagen zij als
onbekend en open. Het enige waar men zo goed als zeker van was, was dat de
revolutie tot bevolkingsdaling zou leiden. Uit de gezaghebbende analyses van
Robert Kuczynski was immers voor experts overduidelijk gebleken dat de vrucht-
baarheid in een groeiend aantal westerse landen onder het vervangingsniveau
lag. Het was alleen nog wachten tot het effect van het positieve bevolkingsmo-
mentum was uitgespeeld (Kuczynski, 1932, 1935; Notestein, 1950). Dit idee uit het interbellum van een demografische revolutie met een open
einde vond later nog weinig bijval. Volgens Dirk van de Kaa (2004) waren het
Princeton-demografen als Frank Notestein, Dudley Kirk en Kingsley Davis die na
de Tweede Wereldoorlog de ‘revolutie’ herdoopten tot een ‘transitie’. Zo schrijft
Kirk (1946, 242): ‘The essence of the vital revolution is the transition from primi-
tive conditions of wasteful mortality and reckless procreation to a new balance
of low death rates and controlled fertility.’ Het verhaal van de ‘demografische
transitie’ was logischer en daardoor erg overtuigend. Het had een duidelijk
einde: het evenwicht van hoge sterfte en vruchtbaarheid ging over in een nieuw
evenwicht van lage sterfte en vruchtbaarheid, want zonder een evenwicht zou
de bevolking ofwel uitsterven ofwel te snel gaan groeien. De logica hiervan werd
bijvoorbeeld heel overtuigend beargumenteerd door Ansley Coale (1986) in het
kader van het Princeton-project over de vruchtbaarheidstransitie in Europa. Inleiding In deze laatste periode zijn chronische ziekten en ondervoeding in
de westerse landen sterk teruggedrongen en kwamen er zeer efficiënte contra-
ceptieve middelen op de markt. Uitstel en afstel van ouderschap werden in veel
grotere mate een kwestie van vrije keuze. Afgezien van mensen die om lichame-
lijke redenen geen kinderen kunnen krijgen, kreeg de kinderloosheid daarmee
fundamenteel een vrijwillig karakter, in tegenstelling tot vroeger (Hakim, 2000;
Poston & Trent, 1982; Dykstra & Hagestad, 2007). In deze bijdrage nemen we de heersende interpretatie van hoge kinderloos-
heid tijdens het interbellum op twee manieren kritisch onder de loep. Ten eer-
ste vatten we samen hoe internationaal vooraanstaande demografen en andere
sociale wetenschappers uit het interbellum de hoge kinderloosheid van hun tijd
interpreteerden. Daaruit zal blijken dat hun visies opvallend verschillen van de
net geschetste redenering. Kort samengevat zagen auteurs uit het interbellum
de volgende oorzaken voor hoge kinderloosheid: een almaar stijgende levens-
standaard, gepaard gaand met een toenemend consumentisme, materialisme en
individualisme en een neiging om de eigen beroeps- en ontspanningsactiviteiten
op de eerste plaats te stellen. We besteden ook aandacht aan de echo’s van het
internationale debat rond de bevolkingskwestie in Nederland, dat door zijn rela-
tief hoge vruchtbaarheid in die jaren een bijzondere positie innam. Ten tweede onderzoeken we hoe individuele en regionale kenmerken ver-
band hielden met de kinderloosheid tijdens het interbellum. Om dit te doen
maken we gebruik van de Historische Steekproef Nederland (Mandemakers, 2000)
en van de Historische Databank Nederlandse Gemeenten (Beekink, Boonstra,
Engelen & Knippenberg, 2003) voor de regionale kenmerken. Waren het vooral
de meer of de minder gegoede beroepsgroepen die kinderloos bleven? Speelde
de godsdienstige denominatie een grote rol? Waren het vooral de vrijzinnige
echtparen die kinderloos bleven? Was de kans op kinderloosheid vooral groot in
regio’s die door hoge werkloosheid getroffen werden? Wat is de samenhang met
het percentage mensen dat bij verkiezingen een stem uitbracht voor een vrijzin-
nige politieke partij? En wat leert ons dit alles over het al dan niet ‘vrijwillige’ en
crisisgebonden karakter van de kinderloosheid tijdens het interbellum? jan van bavel, jan kok en theo engelen Het internationale debat over lage vruchtbaarheid tijdens het
interbellum In La révolution démographique, zowel in het artikel uit 1933 als in het gelijkna-
mige boek uit 1934, schrijft Adolphe Landry daarentegen heel expliciet dat een
belangrijke nieuwigheid van de aan de gang zijnde demografische veranderin-
gen precies het gebrek aan een natuurlijk ‘equilibrium’ is: ‘La remarque capitale,
c’est que dans le régime contemporain, il n’y a plus d’équilibre de la population’ hoge kinderloosheid tijdens het interbellum in nederland (Landry, 1934, 53). Collega’s als Alexander Carr-Saunders (1936) en Enid Charles
(1934) uit Engeland of Alva Myrdal (1941) uit Zweden vielen hem daarin bij. (
)
g
y
(
)
j
Een inherent evenwicht ontbreekt volgens de Franse demograaf, omdat men-
sen tegenwoordig zelf beslissen over het aantal kinderen dat zij willen. Daarmee
is de vruchtbaarheid de speelbal geworden van vaak heel persoonlijke, uiteenlo-
pende en onvoorspelbare overwegingen. Het is dus onmogelijk te zeggen waar de
daling van de vruchtbaarheid zal stoppen (Landry, 1933, 1934). ‘We found reasons
to believe’, aldus zijn Engelse collega Carr-Saunders, ‘that, once the voluntary
small family habit has gained a foothold, the size of the family is likely, if not cer-
tain, in time to become so small that the reproduction rate will fall below replace-
ment rate, and that, when this happened, the restoration of a replacement rate
proves to be an exceedingly difficult and obstinate problem’ (Carr-Saunders, 1936,
327). Tijdens de Tweede Wereldoorlog gingen de prognoses van de Volkerenbond
(voorloper van de Verenigde Naties) inderdaad nog routinematig uit van een con-
tinuering van de voorbije trends, inclusief de verdere daling van de vruchtbaar-
heid, in almaar meer landen tot onder het vervangingsniveau. Ondertussen was
echter vanaf de late jaren dertig en vroege jaren veertig, dus nog tijdens de Twee-
de Wereldoorlog, de babyboom van start gegaan (Notestein, 1950). 54 Motieven voor vruchtbaarheidsbeperking Motieven voor vruchtbaarheidsbeperking In de hedendaagse literatuur wordt de lage vruchtbaarheid uit het interbellum
zoals gezegd vaak geïnterpreteerd als symptomatisch voor economische crisis en
oorlogsdreiging (Frejka & Sardon, 2004, 58; Hakim, 2000; Lesthaeghe & Surkyn,
2004; 2006; Sobotka, 2008). Deze interpretatie steunt niet op empirisch onder-
zoek, noch op het oordeel van vooraanstaande demografen of sociologen uit die
tijd. Volgens Landry (1934) is het juist de gestegen levensstandaard en de daarmee
gepaarde gaande toename van de consumptieaspiraties die mee aanzetten tot
verregaande geboortebeperking. Ook de analyse van Alva Myrdal (1941) gaat in
een compleet andere richting dan het crisisargument: zij argumenteert uitge-
breid dat het goed gaat met de Scandinavische landen in het algemeen en met
Zweden in het bijzonder, dat al jaren vrede kent en dat ook intern geen grote
sociale of etnische conflicten te verwerken heeft. Bovendien laten de geboorte-
cijfers nergens een breuk zien rond 1933. Rond die tijd werd de oorlogsdreiging
in vele Europese landen reëel en ging de angst voor oorlog in toenemende mate
in de hoofden van de mensen spoken (Myrdal, 1941, 7-26). Ook Charles (1934, 81)
minimaliseert het belang van de oorlog voor de ontwikkeling van de vruchtbaar-
heid. De overwegingen die volgens de interbellumauteurs wél aanzetten tot
geboortebeperking zijn heel divers, gaande van het feit dat man en vrouw na
hun huwelijk liever eerst nog wat van het leven genieten (Landry, 1934, 42) tot
de moeilijke combinatie van gezin en arbeid door het onvoldoende aanbod van jan van bavel, jan kok en theo engelen crèches en opvang tijdens de schoolvakanties (Carr-Saunders, 1936, 252; Charles,
1934, 211). Voor een deel gaat het om altruïstische overwegingen in het belang
van de kinderen: als men minder kinderen heeft, dan kan men die met méér
zorg omringen, hen een betere opleiding geven en er voor zorgen dat zij het
verder brengen in het leven dan de ouders zelf. Maar net zo goed spelen vol-
gens Landry (1933, 1934) egocentrische motieven van de ouders een rol. Kinderen
vormen onder andere een beletsel voor de zelfontplooiing van vrouwen buiten
de gezinssfeer. Kinderen vormen een belemmering om vrij aan ontspannings
activiteiten buitenshuis deel te nemen en om te reizen (Charles, 1934; Landry,
1934). Hoewel in hetzelfde huishouden vaak tegelijkertijd egocentrische als
altruïstische motieven spelen, meent Landry (1934, 40-41) dat het gewicht van de
egocentrische overwegingen groter geworden was. Motieven voor vruchtbaarheidsbeperking 55 Zeer gelijkluidende standpunten zijn te lezen bij de Engelse Enid Charles
(1934), die meteen aangeeft dat dergelijke motieven doorgaans hand in hand
gaan met het streven naar meer welvaart en welzijn voor de kinderen: ouders
willen niet alleen voor zichzelf maar vooral ook voor hun kinderen een hogere
levensstandaard. En een vijftal jaar eerder schreef de Duitser Paul Mombert dat
echtparen hun kindertal beperken zowel om het leven voor zichzelf comfortabel
te houden als om hun geliefde kinderen een betere toekomst te bezorgen (Mom-
bert, 1929). Veel later, na de Tweede Wereldoorlog, hebben de Franse auteurs Sauvy
(1960) en Ariès (1980) van de eerste fase van de vruchtbaarheidsdaling een ‘altru-
ïstische’ transitie gemaakt, gevolgd door een tweede, ‘egocentrische’ of ‘indi-
vidualistische’ fase. Deze speculaties over twee opeenvolgende motivaties voor
geboortebeperking hebben duidelijk het denken over een tweede demografische
transitie beïnvloed (zie de verwijzingen naar Sauvy en Ariès in onder andere
Lesthaeghe, 1995; Lesthaeghe & Van de Kaa, 1986). Interpretaties van kinderloosheid Kinderloosheid is volgens gezaghebbende interpretaties uit het interbellum soms
een bewuste keuze (Carr-Saunders, 1936, 256), maar net zo vaak is kinderloos-
heid het gevolg van het almaar op de lange baan schuiven van de stap naar het
ouderschap. Nogal eens lijken echtgenoten te beslissing om na hun huwelijk nog
een tijdje met kinderen te wachten en van het goede leven te genieten. Nadien
blijkt het dan soms moeilijk om de levensstijl waaraan men gewend geraakt is,
op te geven voor kinderen (Landry, 1933, 42). Aan de andere kant van Het Kanaal dacht socioloog en demograaf Alexander
Carr-Saunders er net zo over. De voorbije jaren nam de hoeveelheid vrije tijd en
het aantal mogelijke ontspanningsactiviteiten immens toe, zo schrijft hij. Kinde-
ren vormen een belemmering om daar ten volle van te profiteren (Carr-Saunders,
1936, 111 en 253). Ook Myrdal (1941) argumenteert dat uitstel van ouderschap
voor sommigen een gewoonte is geworden waar soms moeilijk mee te stoppen hoge kinderloosheid tijdens het interbellum in nederland valt. De stap naar het ouderschap veronderstelt dat het echtpaar stopt met het
contraceptieve gedrag waaraan het gewend is geraakt en dat ligt niet altijd voor
de hand: ‘On every single occasion the easiest way is to postpone such a decision’
(Myrdal, 1941, 54). 56 In de volgende alinea’s worden motieven op een rij gezet die volgens gezag-
hebbende demografen uit het interbellum voor uitstel en afstel van ouderschap
zorgden. Secularisatie en rationalisatie Secularisatie en rationalisatie Secularisatie en rationalisatie
In een context waarin contraceptie algemeen ingang heeft gevonden, zijn het
typisch de mensen die vasthouden aan traditionele zeden en gewoonten die nog
een groot kindertal hebben, aldus het oordeel van alle geconsulteerde auteurs. Kinderen zijn een vanzelfsprekendheid voor mensen die veel belang hechten aan
de voorschriften van hun religie en die veel respect opbrengen voor de ‘natuur-
lijke orde’ der dingen (Landry, 1934, 34-37). Uit een analyse van Duitse gegevens
concludeert Mombert (1929, 308-313) dat de vruchtbaarheid vooral in de steden
en in protestantse regio’s sterk beperkt wordt. Dat neemt niet weg dat ook in
katholieke regio’s de daling is ingezet, en ook daar eerst in de steden en pas later
in de rurale gebieden. Dit sterkt Mombert in zijn stelling dat de fundamentele
oorzaak van de lage vruchtbaarheid de rationalisering van het dagelijkse leven
is, die onder andere door de gestegen welvaart en de hogere onderwijsdeelname
in de hand wordt gewerkt (Mombert, 1929, 314-316). ‘La rationalisation de la vie’
is ook voor Landry (1934, 40) de fundamentele verklaring. Kinderloosheid komt
volgens deze auteurs dan ook meer voor bij mensen die zich het moderne ratio-
nele denken eigen hebben gemaakt. De offers van het ouderschap versus de verleidingen van de moderne consumptie
Wie de voor- en nadelen van het ouderschap rationeel afweegt, stuit op een
groot aantal minpunten. Vele van die nadelen waren er ook vroeger al, maar
tegenwoordig wordt er zwaarder aan getild, volgens Myrdal (1941, 53-54). Onder
andere de hedendaagse schoonheidscultus maakt dat sommige vrouwen en hun
echtgenoten vrezen om door zwangerschap en moederschap hun sexappeal te
verliezen. Want een van de opmerkelijke elementen uit de hedendaagse cultuur
is volgens Charles de obsessie met het uiterlijk van vrouwen. ‘Intensive culture of
personal appearance and bodily fastidiousness is not readily reconciled with the
corporal realities of reproduction’ (Charles, 1934, 200). Naarmate het belang van
erotiek in het huwelijk is toegenomen, is voortplanting een minder vanzelfspre-
kende functie van seks tussen echtgenoten geworden. ‘The irruption of children
into the modern erogamic marriage involves a displacement of the emotional
pattern’ (Charles, 1934, 203). Historisch onderzoek naar de evolutie van de huwe-
lijkscultuur wijst inderdaad op een toenemend belang van erotiek en sexappeal jan van bavel, jan kok en theo engelen in huwelijken uit deze periode (Coontz, 2005, Chapter 12). Carr-Saunders (1936,
256) oordeelt dat het huwelijk gevulgariseerd en gedegradeerd is tot ‘a mode of
self gratification’. Secularisatie en rationalisatie Minstens even belangrijk zijn de structurele ontwikkelingen die de sociale
offers van het ouderschap in de snel moderniserende tijden van het interbel-
lum wel erg duidelijk markeren. Zo is niet alleen het beroepswerk maar zijn
ook de moderne recreatieve activiteiten tijdens de toegenomen vrije tijd minder
kindvriendelijk geworden. De hedendaagse ontspanningsindustrie vindt vooral
buitenshuis plaats, aldus Myrdal (1941, 56-58). Ze denkt dan meer bepaald aan
cinema, dancings, ontmoetingsclubs, fietsclubs, picknicks enzovoort. Dergelijke
activiteiten zijn in de regel sterk leeftijdsgebonden, in tegenstelling tot vroeger,
toen generaties hun vrije tijd vaker gemeenschappelijk doorbrachten. Dat geldt
zeker voor het nachtleven, waarvan kinderen uitgesloten zijn, wat er meestal op
uitdraait dat moeder dan maar thuis blijft bij de kinderen. Dit blijft allemaal
niet zonder sociale gevolgen, en vrouwen twijfelen dan ook of ze hun deelname
aan dat soort sociale activiteiten wel willen opgeven voor het moederschap (Myr-
dal, 1941, 58). Voor vrouwen met kinderen is het erg moeilijk er een kleurrijk
sociaal leven op na te houden, zo vat Charles (1934, 197) het samen. 57 p
(
)
Tijdens het interbellum groeide het commerciële aanbod van industrieel
geproduceerde consumptiegoederen en huishoudapparaten explosief. Zo
deden onder andere de stofzuiger, het strijkijzer, de wasmachine, de koelkast,
kindervoeding en luiers hun intrede in het huishouden van de bijdetijdse
middenklasse. Om de verkoop te stimuleren zette de industrie in de periode 1920-
1930 grootschalige moderniseringsoffensieven in. Steeds meer mensen sloten
aan op de water-, gas- en elektriciteitsnetwerken (Buyst, 2006; Schot, 2001). De
geconsulteerde sociaal-demografen merken in dit verband op dat de lat inzake
kinderzorg en hygiëne in het huishouden almaar hoger kwam te liggen (Charles,
1934; Myrdal, 1941). Bovendien hangt aan moderne consumptiegoederen en
huishoudapparaten ook een prijskaartje, om nog maar te zwijgen over de
prijs van een automobiel. Charles merkt in dit verband op: ‘Industrialism has
increased the number of amenities and amusements, more particularly greater
ease of travel, thus providing distractions alternative to parenthood’ (Charles,
1934, 205). Of met een boutade: ‘Statistics clearly show that the choice between
a Ford and a baby is usually made in favour of the Ford’ (Charles, 1934, 197). Wie
het laatste nieuwe snufje graag in huis wilde hebben, stelde daarom misschien
de kinderwens nog wat uit, op het gevaar af dat uitstel in afstel zou uitmonden
(Carr-Saunders, 1936, 252). De moeilijke combinatie van gezin en arbeid De moeilijke combinatie van gezin en arbeid Myrdal (1941, 56) merkt op dat beroepsarbeid in toenemende mate buitenshuis
gebeurt, waardoor betaald werk en zorg voor kinderen moeilijker combineerbaar
worden. Dat dwingt ouders tot het maken van een keuze, waaraan onvermij- hoge kinderloosheid tijdens het interbellum in nederland delijk opportuniteitskosten vasthangen die zeker voor gediplomeerde vrouwen
hoog kunnen oplopen (Myrdal, 1941, 56-59). Kinderen betekenen zonder twij-
fel een rem op de beroepscarrière, beaamt haar Engelse collega-academica, en
moeder van vier kinderen, Charles. Ze noemt een hele reeks van aspecten van
het ouderschap die de carrière van een ambitieuze man of vrouw belemmeren
en besluit: ‘If all scientists were equally intelligent, it is highly probable that
the least fertile ones would achieve the greatest distinction in their profession’
(Charles, 1934, 130-131). 58 Debatten en feiten in Nederland Vlak na de Tweede Wereldoorlog schreef de Amerikaanse demograaf Dudley
Kirk: ‘The Netherlands is the center of an arc of comparatively high fertility that
extends into Germany on the one hand and into Flemish Belgium on the other’
(Kirk, 1946, 49). Dat de lage vruchtbaarheid van vele Europese regio’s tijdens het
interbellum ook wat hem betreft weinig met de economische crisis te maken had,
blijkt onder meer uit wat hij daar onmiddellijk aan toevoegt: ‘The high fertility
of this region is an anomaly in view of its high degree of economic and cultural
development. The Netherlands is the exception to the rule of low birth rates in
Western Europe, an exception not yet adequately explained’ (Kirk, 1946, 49). Dat is een constatering achteraf en vanuit het buitenland, maar hoe dacht
men tijdens het interbellum in Nederland zelf over de demografische situatie? Wanneer men hier te lande over het bevolkingsvraagstuk schreef, dan waren er
in grote lijnen twee opties. In de eerste plaats maakte een deel van de auteurs
zich zorgen over de dreigende overbevolking. Nederland zou maximaal drie tot
vier miljoen mensen kunnen voeden en dus was de situatie (6,7 miljoen inwoners
in 1920) nijpend (Van Praag, 1976, 111). De econoom Coenraed A. Verrijn Stuart
pleitte bijvoorbeeld in diverse publicaties in Economisch Statistische Berichten van
1919 en 1921 voor het tot stilstand brengen van de groei van de bevolking om
te voorkomen dat de economische groei zou worden gefnuikt (Van Praag, 1976,
19-21; zie ook ’t Hooft 1926, 703). De andere, en wellicht nog meer benadrukte betekenis van het bevolkings-
vraagstuk richtte zich op de ethische aspecten van de voortplanting. Bij de oprich-
ting van de Nieuw-Malthusiaanse Bond in 1881 was het nadrukkelijke doel men-
sen in de gelegenheid te stellen de geboorte te voorkomen van kinderen ‘wier
verschijning de kansen op geluk van het gezin vermindert en voor welke gene
vooruitzichten zijn op een menswaardig bestaan’ (geciteerd in Van Praag, 1977,
208). Het openen van dit soort mogelijkheden werd te vuur en te zwaard bestre-
den door de christelijke gemeenschap in Nederland. Volgens Philip van Praag
leidde haar interventie er zelfs toe dat het denken over bevolkingsvraagstukken
in het interbellum gedomineerd werd door zedelijk-normatieve uitgangspunten. Dit bleek bijvoorbeeld uit de steun van hoge zijde bij het nationale congres van
de Vereniging tot bestrijding van het nieuw-malthusianisme in 1919. Debatten en feiten in Nederland Koningin- jan van bavel, jan kok en theo engelen moeder Emma stuurde een financiële bijdrage, terwijl prins Hendrik en diverse
ministers hun steun uitspraken aan de vereniging. Typerend is voorts de discus-
siebijdrage van de medicus en etnoloog Jacob Kohlbrugge uit 1922. Hij vreesde
dat, wanneer ouders eenmaal het kindertal gingen beperken, dit proces snel zou
eindigen bij kinderloze huwelijken. In die zin vond Kohlbrugge een gezonde,
krachtig groeiende bevolking die af en toe geteisterd wordt door hongersnoden
beter dan een gedegenereerde zónder die hongersnoden (Van Praag, 1976, 12,
24-26 en 106). 59 Niet iedereen liet zich meeslepen door de vaak op hoge toon gevoerde debat-
ten. Zo realiseerde Henri Methorst, de latere directeur-generaal van het Centraal
Bureau voor de Statistiek, zich dat er sprake was van een verwarrend proces:
‘We leven in een merkwaardig tijdperk der demografische geschiedenis van ons
land. Terwijl de bevolking door overschot van geboorte boven sterfte sneller dan
ooit toeneemt, zoodat velen zich bezorgd afvragen, waar het heen moet, indien
dit zoo voortgaat, voltrekt zich langzaam een proces, dat in de toekomst voor
onze nederlandsche natie ver strekkende gevolgen kan hebben. We bedoelen:
den achteruitgang van het geboortecijfer, dien we in ons land sedert 1876 aan-
treffen.’ Methorst beseft, met andere woorden, dat de angst voor overbevolking
onterecht is, omdat de toekomstige bevolkingsdaling zich al aankondigt in de
dalende vruchtbaarheid (Methorst, 1914, 3; zie ook Methorst en Sirks, 1948, 33,
77 en 85). Over de redenen achter de daling van het geboortecijfer heeft deze auteur
uitvoerig nagedacht. Hij komt met maar liefst zes redenen. In de eerste plaats
is het zo dat ‘toenemende welstand en beschaving een psychologischen invloed
hebben op ’s menschen wil in de richting van geboorte beperking’. Daar komt bij
dat de democratischer wordende samenleving de mogelijkheid biedt tot sociale
verbetering. In de derde plaats achten sommigen het vermogen tot voortplanten
verminderd door toenemende welstand en beschaving. De trek van het platte-
land naar de stad geldt als vierde reden, die overigens overlapt met de vijfde die
melding maakt van een betere kennis van anticonceptie in steden. Voorts noemt
Methorst nog de daling van de zuigelingensterfte als aanleiding voor geboorte-
beperking (Methorst, 1914, 13-14). Figuur 1 Huwelijksvruchtbaarheid in Nederland 1920-1940 (geboorten per 1000 gehuwde
vrouwen tussen 15 en 45 jaar) Debatten en feiten in Nederland Wat opvalt in de ontwikkeling tij-
dens het interbellum, is dat provincies met een initieel hoge vruchtbaarheid een Figuur 1 Huwelijksvruchtbaarheid in Nederland 1920-1940 (geboorten per 1000 gehuwde
vrouwen tussen 15 en 45 jaar) Figuur 1 Huwelijksvruchtbaarheid in Nederland 1920-1940 (geboorten per 1000 gehuwde
vrouwen tussen 15 en 45 jaar) Bron: Hofstee 1981, p. 132. 0
50
100
150
200
250
300
350
Groningen
Friesland
Drenthe
Overijssel
Gelderland
Utrecht
Noord-Holland
Zuid-Holland
Zeeland
Noord-Brabant
Limburg
NEDERLAND
1921-1925
1926-1930
1931-1935
1936-1940 Bron: Hofstee 1981, p. 132. 0
50
100
150
200
250
300
350
Groningen
Friesland
Drenthe
Overijssel
Gelderland
Utrecht
Noord-Holland
Zuid-Holland
Zeeland
Noord-Brabant
Limburg
NEDERLAND
1921-1925
1926-1930
1931-1935
1936-1940 1921-1925
1926-1930
1931-1935
1936-1940 Bron: Hofstee 1981, p. 132. Bron: Hofstee 1981, p. 132. Debatten en feiten in Nederland Wat ook de precieze oorzaak is voor de daling van het geboortecijfer, Methorst
is zich wel bewust van de grote gevolgen op langere termijn: ‘Terwijl de omlaag
gaande sterftelijn niet altijd kan blijven dalen en vroeg of laat noodzakelijk in
horizontale richting zal moeten ombuigen, zijn aan de daling der geboortelijn
om zoo te zeggen geen grenzen gesteld. Blijven de opvattingen gelden die thans
ten aanzien van het hebben van veel kinderen hoe langer hoe meer in alle lagen
der maatschappij doordringen, dan zal in de toekomst de geboortelijn dalen
tot beneden de sterftelijn, gelijk in Frankrijk reeds in enkele jaren heeft plaats
gehad’ (Methorst, 1914, 27). F.W. ’t Hooft laat in 1936 nog eens duidelijk zien dat groei van de bevolking
kan samengaan met een daling van het geboortecijfer. Hij rekent voor de periode hoge kinderloosheid tijdens het interbellum in nederland hoge kinderloosheid tijdens het interbellum in nederland vanaf 1901 uit dat de groei van de Nederlandse bevolking voor het overgrote deel
is toe te schrijven aan de verlenging van de levensverwachting. Het blijft vreemd,
constateert deze auteur, dat net de periode van grote welvaartsstijging vanaf de
tweede helft van de negentiende eeuw gepaard is gegaan met ‘een verminderden
wil tot procreatie’. Toch is het logisch te verklaren uit het feit dat de sterfte nog
eerder en sterker is gedaald. De daaruit voortvloeiende bevolkingstoename heeft
mensen bang gemaakt: ‘Evenals bij de oude Grieken en bij vele primitieve volken
het dooden van kinderen plaats vond uit angst voor overbevolking, zoo moeten
we de hierboven besproken geboortedaling beschouwen als een reactie op de
snelle bevolkingsvermeerdering. De drijfveer in al deze gevallen is: het pogen
om een eenmaal bereikt welvaartspeil vast te houden’ (’t Hooft, 1936, 100-101 en
109-110). 60 60 Dat was de leer, de overheersende tendens in de literatuur over demografi-
sche onderwerpen. Maar hoe zat het in de werkelijkheid met die dalende vrucht-
baarheid, vooral in het interbellum? Uit figuur 1 wordt meteen duidelijk dat in
alle provincies in elke vijfjaarlijkse periode de vruchtbaarheid lager was dan in
de voorgaande periode. De twee bijna volledig katholieke provincies Limburg en
Noord-Brabant herbergen weliswaar de gehele periode de vruchtbaarste huwe-
lijken, maar ook hier is de daling zonneklaar. Bron: Engelen e.a., 1989. jan van bavel, jan kok en theo engelen jan van bavel, jan kok en theo engelen jan van bavel, jan kok en theo engelen snellere daling laten zien dan de andere provincies. Het gevolg is dat er sprake
is van een duidelijke convergentie van waarden, wat ook blijkt uit de standaard-
deviatie die daalt van 45 via 41 en 37 naar 29. 61 61 De gebruikelijke historische statistieken over vruchtbaarheid bevatten geen
informatie over kinderloosheid. Gelukkig beschikken we over een publicatie die
gebaseerd is op een deel van de volkstelling van 1960. Bij die telling kregen alle op
dat moment nog bestaande echtparen een lijst van vragen voorgelegd over hun
huwelijk, het aantal levendgeboren kinderen en sociale kenmerken (Engelen,
Hillebrand & Van Poppel, 1989). Bij de data zijn verscheidene methodologische
opmerkingen te maken, onder meer over de selectiviteit van de overlevenden in
1960 in termen van hun vruchtbaarheid. Niettemin zijn de gegevens bruikbaar
om een eerste blik te kunnen werpen op de kinderloosheid van overlevende stel-
len die trouwden tijdens het interbellum. Voor het bijgaande overzicht kozen wij
twee huwelijkscohorten (1919-1923 en 1934-1938) uit verschillende regio’s bin-
nen Nederland (stedelijk, half stedelijk en ruraal)2, in totaal bijna 50.000 huwelij-
ken. Voor al deze huwelijken is verder een differentiatie mogelijk naar geloof en
beroepsgroep (tabel 1). De eerste conclusie uit tabel 1 moet luiden dat de stijging van het aandeel
kinderloze huwelijken over de twee huwelijkscohorten erg beperkt is, althans
bij de huwelijken die tot 1960 overleefden. De andere uitkomsten liggen in de
lijn van de bevindingen uit eerder onderzoek naar de huwelijksvruchtbaarheid
tijdens het interbellum in Nederland (Van Poppel, 1983) en van een interpretatie Tabel 1 Percentage kinderloze huwelijken in Nederland in 1960, huwelijkscohorten
1919-1923 en 1934-1938
%
N
%
N
Totaal
7,6
49.626
Beroep
Landbouw
5,1
8.353
Cohort
Arbeiders
6,3
17.781
1919-1923
7,6
21.165
Zelfstandigen
7,9
7.648
1934-1938
7,7
28.461
Witte boorden
9,8
9.233
Religie
Regio
Katholiek
6,8
9.933
Rotterdam
9,5
24.273
Nederlands Hervormd
7,4
20.416
Urbaan platteland NH
7,3
4.414
Gereformeerd
5,9
6.653
Nijmegen
6,2
3.390
Andere religie
8,8
2.246
Platteland Utrecht
6,0
2.200
Geen religie
9,7
10.378
Platteland Friesland
6,0
9.766
Platteland Overijssel
4,4
5.583
Bron: Engelen e.a., 1989. jan van bavel, jan kok en theo engelen Tabel 1 Percentage kinderloze huwelijken in Nederland in 1960, huwelijkscohorten
1919-1923 en 1934-1938 hoge kinderloosheid tijdens het interbellum in nederland hoge kinderloosheid tijdens het interbellum in nederland van kinderloosheid in termen van secularisatie en rationalisatie: de hoogste kin-
derloosheid werd opgetekend bij de paren die niet religieus zeiden te zijn, ter-
wijl de laagste cijfers te vinden zijn bij de meest behoudende religies, namelijk
de gereformeerden en rooms-katholieken. 62 62 Wat de beroepsgroepen betreft is het geen verrassing dat de landbouw, die de
meest vruchtbare beroepsgroep vormt, ook weinig kinderloze paren telt. Naar-
mate men opklimt op de sociale ladder neemt het percentage kinderlozen toe. Binnen de witte boorden nemen de vrije beroepen met 12,5 procent overigens
de toppositie in. De regionale spreiding wijst uit dat in het verstedelijkte westen
van het land kinderloosheid relatief vaak voorkomt, maar ook op het platteland
zijn in provinciale stedelijke centra weer meer echtparen kinderloos dan buiten
de steden (vergelijk Van Poppel, 1983). Data en hypothesen Volgens de interbellumdemografen had kinderloosheid bij echtparen onder meer
te maken met de verminderde invloed van godsdienstige overwegingen, met een
toegenomen consumptiedrang, met het toegenomen aanbod van kindonvrien-
delijke ontspanningsactiviteiten en met de moeilijker wordende combinatie van
beroepsarbeid met moederschap. De economische crisis van de jaren dertig en
de oorlogsdreiging vanaf 1933 speelden in hun ogen een minder grote rol. De
vraag is in welke mate deze contemporaine opvattingen bevestigd of weerlegd
worden door de statistische gegevens. Om dat na te gaan, steunen we enerzijds
op gezinsreconstructies op basis van de Historische Steekproef Nederland (HSN),
waaruit we kenmerken van individuele echtparen distilleren, en anderzijds op
contextuele informatie. Al naargelang het wenselijk en mogelijk is, brengen we
contextuele gegevens op het gemeentelijke en op het regionale niveau in reke-
ning. Voor de indeling in regio’s is gebruikgemaakt van de economisch-geografi-
sche regionale classificatie uit de beroepstelling van 1930, die Nederland indeelt
in 42 gebieden met een min of meer homogene economische structuur. Zo is
regio 1 de Noord-Groningse Bouwstreek en regio 42 de Mijnstreek in Limburg. Alle gebruikte contextuele gegevens zijn afkomstig uit de Historische Databank
Nederlandse Gemeenten (HDNG; Beekink e.a., 2003). Kinderloosheid op basis van HSN Voor het onderzoek naar de achtergronden van kinderloosheid hebben we een
dataset opgebouwd met huwelijken gesloten tussen 1919 en 1938. Hiervoor is
gewerkt met de data van de Historische Steekproef Nederlandse bevolking (HSN), data-
set Levenslopen, release 2007.01 (zie de bijdrage van Mandemakers in dit boek).3 Het
gaat bij de selectie om eerste huwelijken, waarvan de exacte huwelijksdatum
bekend was. Omdat de burgerlijke staat bij huwelijk niet in het bevolkingsregis-
ter is vermeld, is hierbij ook gebruikgemaakt van de collectie huwelijksakten jan van bavel, jan kok en theo engelen van de HSN (HSN dataset Akten burgerlijke stand 2007.01). Het maken van gezins-
reconstructies wordt enigszins bemoeilijkt door de persoonskaarten, die vanaf
1940 in de plaats kwamen van de gezinskaarten. Alle personen kregen toen een
eigen kaart, maar de kinderen die op 1 januari 1940 nog thuis woonden, werden
ook op de kaart van het gezinshoofd bijgeschreven (Vulsma, 2002). Echter, het
gezinshoofd was vrijwel altijd de man. Enkel als de man kwam te overlijden of
wanneer een scheiding werd uitgesproken, werden de nog levende kinderen op
de kaart van de vrouw bijgeschreven. Als de HSN-onderzoekspersoon een vrouw
is, hebben we dus de kaart van haar man nodig om de gezinsreconstructie vanaf
1940 te kunnen vervolgen. Niet in alle gevallen is de HSN er in geslaagd de per-
soonskaarten van echtgenoten van vrouwelijke onderzoekspersonen te verwer-
ven. Wanneer dit betekent dat een huwelijk niet minstens vijftien jaar kon wor-
den gevolgd (zie onder), zijn deze gevallen buiten het onderzoek gelaten. 63 Huwelijken die werden afgesloten op een moment dat de vrouwelijke part-
ner al veertig jaar of ouder was, zijn uit de analyse geweerd, omdat haast al deze
huwelijken kinderloos bleven, wellicht om fysiologische redenen. Voor sommige
huwelijken ontbraken kleine stukjes informatie, doordat registratiegegevens
niet werden teruggevonden. In de analyse wordt daarmee rekening gehouden
door te kijken of het aantal maanden ontbrekende informatie samenhangt met
de kans om als kinderloos huwelijk in de HSN-data op te duiken. Kinderloosheid Dit onderzoek heeft als afhankelijke variabele de kinderloosheid van huwelij-
ken en niet hun steriliteit. Het gaat over de nettoreproductie en niet om de lou-
tere vruchtbaarheid, dus enkel overlevende kinderen worden meegeteld. Meer
bepaald beschouwen we een huwelijk als definitief kinderloos als er vijftien jaar
na de trouwdatum geen kind uit dat huwelijk in leven is. De grens lager leggen,
bijvoorbeeld na tien jaar huwelijk, heeft als nadeel dat een niet verwaarloosbaar
deel nadien nog een kind krijgt. Met name bij de vroeg getrouwden is dat het
geval: vrouwen die trouwden vóór de leeftijd van twintig jaar en die na tien jaar
huwelijk nog kinderloos waren, kregen in een aanzienlijk deel van de gevallen
nog een kind vóór ze vijftien jaar getrouwd waren. De grens voor ‘definitieve’
kinderloosheid hoger leggen, bijvoorbeeld na twintig jaar huwelijk, is niet wen-
selijk, omdat de selectiviteit dan sterker begint te spelen, vooral als gevolg van
huwelijksontbinding door sterfte. Bovendien bleek uit onze proefanalyses dat
het uiterst zelden voorkomt dat een kinderloos huwelijk na vijftien jaar alsnog
een kind voortbrengt. Na toepassing van alle vermelde selectiecriteria bleven 2884 huwelijken over
voor analyse van kinderloosheid. Het gaat dus om eerste huwelijken gesloten ná
1918 maar vóór 1940 die minstens vijftien jaar intact bleven. Door ontbrekende
waarden bij verklarende variabelen wordt het aantal observaties waarop gesteund
wordt in de regressieanalyses soms beperkt tot minimaal 2782 huwelijken. hoge kinderloosheid tijdens het interbellum in nederland Tabel 2 geeft per huwelijkscohort het percentage dat na vijftien jaar zon-
der overlevend kind is. De cijfers geven aan dat dit percentage steeg van 13 tot
19 procent over de beschouwde cohorten. Deze schattingen liggen aanzienlijk
hoger dan wat verwacht zou worden op basis van tabel 1. Daar zijn verschillende
verklaringen voor mogelijk die elk tegelijk een deel van de waarheid kunnen
bevatten. Ten eerste is tabel 1 gebaseerd op het (bruto-)aantal levendgeborenen
per bestaand huwelijk, en tabel 2 op nettoreproductiecijfers. De zuigelingen-
sterfte lag tijdens het interbellum nog redelijk hoog: rond de 50 per 1000 levend-
geborenen. Tegen het einde van de Tweede Wereldoorlog piekte dit cijfer zelfs
kortstondig tot rond de 70 per 1000 (NIDI, 2003, 96). Met de beschikbare HSN-
release is het niet mogelijk om op betrouwbare wijze na te gaan wat de impact
van zuigelingensterfte op de percentages eerste huwelijken zonder overlevende
kinderen is. Kinderloosheid Maar een deel van de stijging van de kinderloosheid moet wellicht
op het conto van de verhoogde zuigelingensterfte tijdens de Tweede Wereldoor-
log geschreven worden. Een tweede en wellicht belangrijker verklaring is dat de
cijfers in tabel 1 enkel gelden voor echtparen die overleefden tot 1960. Onder-
zoek heeft al vaak uitgewezen dat de sterfte bij kinderloze vrouwen aanzienlijk
hoger ligt dan bij vrouwen met kinderen, vooral als die kinderen zelf niet vroeg-
tijdig sterven (Doblhammer, 2000; Hurt, Ronsmans, Campbell, Saha, Kenward
& Quigley, 2004; Kumle & Lund, 2000; Manor, Eisenbach, Israeli & Friedlander,
2000). Volgens Amerikaans onderzoek was er in deze periode ook een sterk ver-
hoogde kans op echtscheiding bij kinderloze echtparen (Jacobson, 1950). Dat er
in de hier geanalyseerde HSN-data aanzienlijk meer kinderloze echtparen zitten,
wordt dan ook voor een deel verklaard door het feit dat de HSN ook echtparen
telt die het jaar 1960 niet zullen halen. Een derde mogelijke verklaring is dat het
aandeel kinderloze paren in de HSN overschat wordt, bijvoorbeeld omdat over-
levende kinderen ten onrechte niet geregistreerd of gevonden werden. Nochtans
hebben we gevallen waar we een verhoogd gevaar liepen om het spoor van even-
tuele kinderen te missen, systematisch niet in de analyse opgenomen. Bovendien
hebben de HSN-schattingen dezelfde orde van grootte als bijvoorbeeld die van
Anderson (1998) voor Groot-Brittannië op basis van de Family Census van 1946. Volgens de Britse gegevens bleef ongeveer 16 procent van de huwelijken uit het
jaar 1925 kinderloos. Wij komen voor de huwelijkscohorten 1924-1928 op basis
van HSN uit op hetzelfde percentage. In de Verenigde Staten van Amerika lagen
de cijfers nog een stuk hoger (Morgan, 1991). Daarom achten we het onwaar-
schijnlijk dat de HSN het aantal kinderloze paren overschat 64 Hypothesen en operationalisering van variabelen Godsdienstige gezindte en het regionale ideologische klimaat g g
g
g
De meeste auteurs vermelden de secularisatie en de rationalisering van de per-
soonlijke levenssfeer als een factor die het kiezen voor een leven zonder kind ver- jan van bavel, jan kok en theo engelen gemakkelijkt (Charles, 1934; Landry, 1933; Mombert, 1929; Myrdal, 1941). Hieruit
worden twee hypothesen afgeleid. Ten eerste zou men verwachten dat echtparen
die zelf blijk geven van een eerder ‘rekkelijke’ godsdienstige gezindte een hogere
kans maken om kinderloos te blijven. Ten tweede zou men ook meer kinderloos-
heid verwachten bij echtparen die leven in een regio waar een groot deel van de
populatie eerder progressieve en geseculariseerde opvattingen heeft. 65 De bevolkingsregisters vermelden de godsdienstige gezindten van man en
vrouw. We zien af van eventuele veranderingen in gezindte door de tijd heen en
gebruiken voor onze analyse de eerste vermelding gerekend vanaf het huwelijk. Onze indeling van denominaties is gebaseerd op de expliciete stellingnamen van
de betreffende kerken in de discussies over geboortebeperking en de ruimte die
aan individuele kerkleden werd gelaten om hun eigen geweten en oordeel te vol-
gen (zie Kok & Van Bavel, 2006). Naast de rooms-katholieken onderscheiden we
‘orthodox-protestanten’, die bestaan uit alle groepen gereformeerden, de ‘evan-
gelischen’ (baptisten, leden van het Leger des Heils et cetera) en de (vermoede-
lijke) aanhangers van de Gereformeerde Bond en de Confessionele Vereniging
binnen de Nederlands Hervormde Kerk. De aanhangers van de laatstgenoemde
orthodoxe stromingen zijn niet als zodanig in het bevolkingsregister geregi-
streerd. Ze verschillen qua mentaliteit echter zó sterk van de ‘ethisch’ en ‘vrijzin-
nig’ Nederlands Hervormden dat we gekozen hebben voor de volgende oplossing. Wanneer deze hervormden waren geboren in een plaats waar alle predikanten
in 1920 behoorden tot de Gereformeerde Bond of de Confessionele Vereniging,
is besloten ze bij de orthodoxen onder te brengen (Knippenberg, 1992, 106-114;
Kok en Van Bavel, 2006). Onder de ‘liberaal-protestanten’ rekenen we Waals en
Nederlands Hervormden (uitgezonderd de zojuist genoemden), de evangelisch-
lutherschen, de remonstranten en de doopsgezinden. Aparte groepen worden
gevormd door joden en onkerkelijken. Vervolgens combineerden we de denomi-
naties van de mannelijke met die van de vrouwelijke echtgenoot en hergroepeer-
den we volgens de indeling in tabel 2. Opvallend daarbij is dat de huwelijken
met beide partners liberaalprotestanten, duidelijk in aandeel verminderden in
de eerste helft van vorige eeuw, van 31 naar 21 procent. Hypothesen en operationalisering van variabelen Het aandeel huwelijken
waarbij een protestantse man of vrouw huwde met een vrouw of man zonder
denominatie nam sterk toe, alsook het aandeel huwelijken waarbij hij noch zij
een godsdienstige denominatie invulde. Wegens hun pronatalistische ideologie verwachten we de minste kinderloos-
heid bij homogeen rooms-katholieke echtparen en bij orthodox-protestanten. Een hogere kans op kinderloosheid verwachten we bij gemengde huwelijken,
juist omdat het religieus gemengd karakter van een zeker rekkelijkheid getuigt. Ook bij eerder liberaal-protestantse denominaties wordt een hogere kans op kin-
derloosheid verwacht. De hoogste kans op kinderloosheid is er naar verwachting
bij de echtparen zonder godsdienstige denominatie. Op het regionale niveau krijgen we een indruk van het ideologische klimaat
op basis van het percentage stemmen op progressieve partijen bij de Tweede hoge kinderloosheid tijdens het interbellum in nederland Tabel 2 Beschrijvende statistiek over de gebruikte variabelen Huwelijksjaar
Huw.-
1919-23 1924-28 1929-33 1934-38 Totaal
leef
tijd
vrouw
N
899
793
714
478
2.884
Kinderloos na 15 jaar (%)
12,9
16,3
15,6
19,3
15,5
Huwelijksleeftijd vrouw (M)
24,9
25,1
24,8
25,4
25,0
(s.d.)
(4,3)
(4,3)
(4,6)
(4,5)
(4,4)
Huwelijksleeftijd man (M)
27,0
27,4
27,2
27,5
27,2
(s.d.)
(4,9)
(5,4)
(5,1)
(4,6)
(5,0)
Beroep man (%)
Ongeschoolde arbeiders, incl. Hypothesen en operationalisering van variabelen landarbeiders
21,7
19,9
22,1
19,2
20,9
24,0
Professionals en economische elite
2,1
3,2
2,0
3,3
2,6
26,3
Witte boorden
21,1
21,6
20,2
22,8
21,3
25,6
Ambachtslui en laag- tot hooggeschoolde
arbeiders
38,4
35,9
38,8
40,6
38,2
24,8
Boeren, vissers en geschoolde landarbeiders
14,0
16,0
12,9
10,0
13,6
26,0
Geen beroepsvermelding
2,7
3,4
4,1
4,0
3,4
25,0
Beroepsvermelding vrouw (%)
Geen vermelding
73,4
76,8
81,1
84,1
78,0
24,7
Geschoold
3,2
3,4
2,4
1,3
2,7
29,7
Ongeschoold
23,4
19,8
16,5
14,6
19,2
25,4
Godsdienstige gezindte (%)
Beide rooms-katholiek
23,6
20,3
23,7
21,5
22,4
25,7
Beide liberaal-protestants
31,4
29,6
25,4
21,5
27,8
24,7
Beide orthodox-protestants
7,0
9,0
8,1
6,1
7,7
24,7
Hij katholiek, zij protestants
3,6
4,3
3,1
1,5
3,3
25,4
Hij protestants, zij katholiek
4,7
5,2
2,9
3,3
4,2
24,4
Gemengd protestants
13,3
11,2
10,1
10,7
11,5
25,2
Gemengd katholiek – geen/onbekend
3,9
3,9
5,0
10,5
5,3
25,2
Gemengd protestants – geen/onbekend
10,6
12,9
18,3
20,5
14,8
24,7
Joods
1,3
1,0
0,8
0,4
1,0
25,2
Beide geen of onbekend
0,7
2,6
2,5
4,0
2,2
24,0
Gemeentelijke kenmerken
Levensstandaard/consumptie (factorscores)
Nutsvoorzieningen (1934-35)
-0,04
Aanbod winkels (1930)
0,01
Bezit auto’s en motoren (1935)
-0,01
Regionale kenmerken
% Werkloze mannen (1933)
14,1
% Progressieve stemmen (1937)
36,5
Bronnen: HSN-datasets Levenslopen en Akten, release 2007.01 en Historische Databank Nederlandse Gemeenten. Huwelijksjaar 66 nnen: HSN-datasets Levenslopen en Akten, release 2007.01 en Historische Databank Nederlandse Gemeenten. jan van bavel, jan kok en theo engelen Kamerverkiezingen van 1937. Als progressieve partijen zijn aangemerkt de Com-
munistische Partij Nederland, de Sociaal-Democratische Arbeiderspartij, De Vrij-
heidsbond en de Vrijzinnig Democratische Bond. Hoe hoger het percentage pro-
gressieve stemmen in een regio, hoe hoger de verwachte kans op kinderloosheid. 67 Beroepspositie en werkloosheid De laatste kolom van tabel 2 laat zien dat vrouwen met een geschoolde
beroepspositie een aanzienlijk hogere gemiddelde huwelijksleeftijd hebben hoge kinderloosheid tijdens het interbellum in nederland dan vrouwen zonder geregistreerd beroep of vrouwen met een ongeschoolde
beroepspositie. Dat geeft extra aanleiding tot de hypothese dat er in de groep
van geschoolde vrouwen meer kinderloze huwelijken zullen zijn dan bij de rest. De vraag is dan of een eventueel verhoogde kinderloosheid ook nog standhoudt
als rekening wordt gehouden met die late intrede in het huwelijk. 68 68 De economische crisis en werkloosheid van de jaren dertig figureert minder
prominent in de toenmalige literatuur als verklaring van lage vruchtbaarheid
dan economische vooruitgang. Niettemin onderzoeken we de regionale invloed
van de depressie via het percentage werklozen op de mannelijke beroepsbevol-
king in 1933, gemeten op het niveau van de 42 geografisch-economische regio’s. Op het individuele niveau zullen werklozen tot op zekere hoogte terug te vinden
zijn in de categorie van mannen zonder beroepsvermelding. De hypothese luidt
dat er een hogere kans op kinderloosheid is in regio’s met een hoge werkloos-
heid in 1933 en bij mannen zonder beroepsvermelding. Consumptieniveau en moderne commercie
Kinderloosheid zou volgens zowat alle geconsulteerde interbellumauteurs
samenhangen met de toegenomen consumptiedrang, die als zo kenmerkend
voor de moderne tijden werd beschouwd. Op het niveau van de individuele echt-
paren zijn er echter geen indicatoren voor het aantal aangekochte consumptie-
goederen. In de HDNG zijn er echter wél indicatoren op het gemeentelijke niveau
beschikbaar. Op basis daarvan toetsen we de hypothese dat in gemeenten waar
klaarblijkelijk een hoog ‘modern’ consumptieniveau heerste, echtparen gemid-
deld ook een hogere kans op kinderloosheid hadden. De gebruikte HDNG-indicatoren zijn (met telkens het referentiejaar er tussen
haakjes bij): het aantal aansluitingen op het elektriciteitsnet (1934), het aantal
aansluitingen op het gasdistributienet (1934), het aantal telefoonaansluitingen
(1935), het geregistreerde aantal radiotoestellen (1935), het aantal personenau-
to’s (1935) en het aantal motorvoertuigen (1935), het aantal voedingswinkels,
kledingwinkels en winkels in huishoudelijke artikelen (telkens in 1930). Al deze
cijfers worden uitgedrukt per 100 inwoners in de betreffende gemeente in 1934. Om de samenhang tussen deze indicatoren samen te vatten in een of enkele
latente variabelen, werd gebruikgemaakt van exploratieve factoranalyse (meer
bepaald iterated principal axis factoring met varimax rotatie) (Kim & Mueller, 1978). Tabel 3 geeft het resultaat van deze factoranalyse. Beroepspositie en werkloosheid Op basis van de literatuur uit de eerste helft van de twintigste eeuw zou men
verwachten dat vrijwillige kinderloosheid vooral voorkwam in de geschoolde
sociale rangen. Daarom verwachten we dat de kans op kinderloosheid het hoogst
is bij professioneel geschoolden en witteboordenberoepen. Lage kinderloosheid
wordt verwacht bij boeren en ongeschoolde arbeiders. Deze verwachtingen gel-
den met betrekking tot de mannenberoepen, maar a fortiori met betrekking tot
de vrouwenberoepen. Helaas is de registratie van de vrouwenberoepen naar onze
mening vrij onbetrouwbaar en onder meer afhankelijk van de opvattingen van
de lokale ambtenarij (sommige ambtenaren waren wellicht meer geneigd om
voor gehuwde vrouwen een beroep te noteren dan andere). Voor de overgrote
meerderheid van de gehuwde vrouwen is er trouwens geen enkele beroepsver-
melding beschikbaar. Daarom beperken we ons noodgedwongen tot een driede-
ling: vrouwen zonder beroep, vrouwen met een beroep waarvoor enige scholing
wordt verwacht en vrouwen met een beroep waarvoor geen scholing vereist is. Net als bij de meting van religieuze gezindte is ook bij de beroepen geko-
zen voor één momentopname, zodat beroepsmobiliteit hier buiten beschouwing
blijft. In eerste instantie nemen we voor de echtelieden de beroepen voor zover
vermeld op de huwelijksakte. Indien de akte ontbreekt, nemen we het eerstge-
noemde beroep na het huwelijk in het bevolkingsregister. Alle mannenberoepen
zijn gecodeerd in HISCO (Van Leeuwen, Maas & Miles, 2002) en geclassificeerd in
sociale groepen met HISCLASS (Van Leeuwen, Maas & Miles, 2005). Daarbij zijn
de twaalf HISCLASS-groepen teruggebracht tot zes beroepscategorieën, waarvan
één voor mannen waarvoor een beroepsvermelding ontbrak (zie tabel 2). Om het aantal vrouwen met beroepsvermelding te maximaliseren, hebben
we bij vrouwen die na hun huwelijk zonder beroep geregistreerd stonden, ook
het laatst vermelde beroep vóór het huwelijk ingevuld. De onderliggende werk-
hypothese is dat eventuele voorhuwelijkse beroepservaring (ook) van invloed
is op de latere reproductie. Laag- of ongeschoolde (land)arbeiders en (winkel)
bedienden (HISCLASS-categorieën 5, 9, 10, 11 en 12, zie Van Leeuwen e.a., 2005)
beschouwen we als ongeschoold, de overige categorieën als geschoold. Ondanks
de ruimdenkendheid in het toekennen van beroepsposities aan vrouwen, blijft
de overgrote meerderheid zonder beroepsvermelding. Over de huwelijkscohor-
ten bekeken neemt het aandeel vrouwen zonder geregistreerd beroep zelfs nog
toe van 73 tot 84 procent (zie tabel 2). Historische trend en uitstel van huwelijk De internationale literatuur uit het interbellum maakt gewag van een toene-
mende neiging van echtparen om voor een leven zonder kinderen te kiezen. Een
zijdelingse vraag is of daar ook in Nederland aanwijzingen voor zijn. De cijfers in
tabel 1 suggereren dat er hooguit van een beperkte stijging sprake zou zijn. Wat
zegt de analyse op basis van HSN? En als er al sprake zou zijn van een diachrone
trend, valt die dan te verklaren door een eventuele stijging van de huwelijksleef-
tijd tijdens deze periode? Tot nu toe was het interbellum een blinde vlek in de
tijdreeks van huwelijksleeftijden voor Nederland (zie NIDI, 2003, 80; Van Poppel,
1992). De volgende analyses brengen zowel de huwelijksleeftijd van de man als
die van de vrouw in rekening. Beroepspositie en werkloosheid Er werden drie factoren geëx-
traheerd, omdat de eigenwaarde vanaf een vierde factor onder de grens van één
dook. De (betekenisgevende) hoge factorladingen zijn vetgedrukt. Op basis van
deze analyse werd voor elke Nederlandse gemeente voor elk van de drie factoren
een gestandaardiseerde score berekend, die mee in de regressieanalyses wordt
opgenomen. De eerste factor correleert sterk met de aansluitingen op gas, elektriciteit,
telefoon en met het aantal radiotoestellen. Deze factor krijgt daarom de naam jan van bavel, jan kok en theo engelen ‘nutsvoorzieningen’. De tweede factor staat duidelijk voor het gemeentelijke
aanbod van diverse soorten van winkels. De laatste factor vat vooral het bezit van
auto- of motorvoertuigen samen. 69 Historische trend en uitstel van huwelijk Historische Databank Nederlandse Gemeenten. actiemethode: iterated principal axis factoring; rotatiemethode: varimax. Resultaten In wat volgt wordt de kans op nul overlevende kinderen na vijftien jaar huwelijk
gemodelleerd met behulp van een mixed effects multilevel logistisch-regressiemo-
del. Om de parameters te schatten maakten we gebruik van de Laplace-schattings-
techniek, die in de literatuur als preciezer beschreven staat dan meer gebruike-
Tabel 3 Exploratieve factoranalyse van gemeentelijke indicatoren voor consumptieniveau:
factorladingena Factor 1
Factor 2
Factor 3
‘Nutsvoorzieningen’
‘Winkels’
‘Auto/motor’
Elektriciteitsaansluitingen
0,71
-0,10
0,28
Gasaansluitingen
0,79
0,14
0,22
Telefoonaansluitingen
0,64
0,07
0,45
Radiotoestellen
0,76
0,00
0,08
Personenauto’s
0,20
0,12
0,81
Motorvoertuigen
0,26
0,06
0,67
Voedingswinkels
-0,19
0,66
-0,03
Kledingwinkels
0,32
0,87
0,04
Winkels huishoudelijke artikelen
0,04
0,79
0,25
Eigenwaarde
2,37
1,87
1,50 a Extractiemethode: iterated principal axis factoring; rotatiemethode: varimax. hoge kinderloosheid tijdens het interbellum in nederland lijke alternatieven (Agresti, 2002; Snijders & Bosker, 1999).4 Er worden gefixeerde
effecten (fixed effects) geschat voor verklarende variabelen op drie niveaus: het
niveau van individuele echtparen, het niveau van de gemeente waarin zij wonen
en het niveau van de regio waarin die gemeente zich bevindt. Als woonplaats
nemen we de eerste vestigingsplaats na het huwelijk, ook voor echtparen die
nadien verhuisden. Daarnaast worden random intercepts geschat op het regionale
niveau, wat impliceert dat het model er rekening mee houdt dat het gemiddelde
niveau van kinderloosheid varieert als gevolg van onbekende kenmerken van de
regio. Het was niet mogelijk om ook betrouwbare random effects op het gemeente-
lijke niveau te schatten, omdat voor vele gemeenten slechts één representant in
de HSN-data aanwezig is. 70 70 De verklarende variabelen worden stapsgewijs toegevoegd: eerst bekijken we
de effecten van variabelen op het individuele niveau, dan die van het regionale en
het gemeentelijke niveau, en ten slotte gaan we na wat de netto-, directe effecten
zijn als alle variabelen tegelijkertijd aan het model worden toegevoegd (tabel 4). Zijn mannen en vrouwen met een geschoolde beroepspositie vaker kinder-
loos dan ongeschoolden? Zijn echtparen met een eerder progressieve godsdien-
stige gezindte meer geneigd om kinderloos te blijven? En zo ja, valt dat te ver-
klaren door de hogere huwelijksleeftijd van de vrouw in kwestie of gelden de
verschillen ook na controle voor de huwelijksleeftijd? De vergelijking tussen de
modellen I en II in tabel 4 geeft antwoorden op deze vragen. Wat de beroepsgroep van de mannelijke partner betreft, is de kans op kinder-
loosheid volgens model I veruit het hoogst bij de mannen zonder beroepsvermel-
ding. Resultaten Wellicht gaat het hier in vele gevallen om mannen die om economische of
gezondheidsredenen al dan niet langdurig werkloos zijn. Dat kan gezien worden
als een aanwijzing dat een economisch precaire situatie aanleiding gaf tot afstel
van ouderschap. De groep met de tweede grootste kans op kinderloosheid is die
van de witte boorden. De vergelijkingsgroep is telkens die van de ongeschoolde
arbeiders. Ook geschoolde arbeiders, boeren en vissers hebben een significant
hogere kans op kinderloosheid dan de vergelijkingsgroep, althans wanneer de
huwelijksleeftijd niet in rekening wordt gebracht. Hooggeschoolde professionals
en managers hebben eveneens een aanzienlijk hogere kans op kinderloosheid
volgens model I, maar de standaardfout van dit effect is groot, wellicht door
de kleine aantallen in deze groep. Wat vrouwenberoepen betreft hebben de
geschoolde vrouwen een duidelijk hogere kans op kinderloosheid dan vrouwen
zonder geregistreerd beroep. De verschillen tussen de beroepsgroepen verkleinen echter, in sommige
gevallen fors, wanneer ook de huwelijksleeftijd in rekening wordt gebracht
(model II). Bij de vrouwen is het verschil tussen de geschoolde beroepen en de
vrouwen zonder beroep dan bovendien niet meer statistisch significant. Bij de
mannen blijft de hoge kinderloosheid bij de geschoolde arbeiders, de witte boor-
den en vooral bij de mannen zonder beroepsvermelding overeind. De eerder
gesignaleerde relatief hoge kinderloosheid bij zowel de hooggeschoolde profes- jan van bavel, jan kok en theo engelen sionals en managers als de landbouwers en vissers blijkt vooral een gevolg te zijn
van hun gemiddeld hoge huwelijksleeftijd. De verschillen tussen de religieuze denominaties worden eerder sterker dan
zwakker wanneer met de huwelijksleeftijd rekening wordt gehouden. Liberaal-
protestanten hebben een gemiddeld lage huwelijksleeftijd en verschillen niet
significant van de referentiegroep van katholieke huwelijken als daar niet voor
gecontroleerd wordt (model I). Wanneer hun gemiddeld vroege huwelijksintre-
de wél in rekening wordt gebracht, stijgt het nettoverschil met de referentie-
groep aanzienlijk en wordt het statistisch significant: de kans om na vijftien jaar
huwelijk geen overlevende kinderen te hebben in verhouding tot de kans om wél
overlevende kinderen te hebben, is bij huwelijken tussen twee liberaal-protes-
tanten ongeveer 64 procent hoger dan bij huwelijken tussen twee katholieken
(exp(0,497) = 1,64). Een gelijkaardig verhaal is van toepassing op de gemengde
huwelijken, hetzij katholiek met protestants, hetzij katholiek of protestants met
een partner zonder godsdienstige denominatie: zeker wanneer de huwelijksleef-
tijd in de vergelijking wordt opgenomen, hebben deze ‘rekkelijke’ gezindten een
duidelijk hogere kans op kinderloosheid dan de katholieken. Resultaten Intercept
-50,490 ** 19,230
-46,330 *
19,860
-58,790 **
18,800
-58,170 ** 19,000
-49,700 *
20,030
Huwelijksjaar
0,025 *
0,010
0,021 *
0,010
0,027 **
0,010
0,027 **
0,010
0,023 *
0,010
Huwelijksleeftijd vrouw
0,120 ***
0,016
0,111 *** 0,015
0,109 ***
0,015
0,118 ***
0,016
Huwelijksleeftijd man
0,025 ~
0,013
0,026 *
0,013
0,028 *
0,013
0,026
0,014
Aantal ontbrekende maanden
0,001
0,001
0,000
0,001
0,001 ~
0,001
0,001
0,001
0,000
0,001
Beroep man
Ongeschoolde arbeider (ref.)
0,000
/
0,000
/
0,000
/
Professionals & managers
0,454
0,357
0,192
0,368
0,011
0,382
Witte boorden
0,660 ***
0,180
0,502 **
0,186
0,418 *
0,191
Geschoolde arbeiders
0,414 *
0,167
0,346 *
0,173
0,269
0,177
Boeren en vissers
0,497 *
0,209
0,189
0,219
0,333
0,223
Geen beroepsvermelding
2,146 ***
0,260
2,116 ***
0,269
2,054 ***
0,274
Beroepsvermelding vrouw
Geen beroepsvermelding (ref.)
0,000
/
0,000
/
0,000
/
Geschoold
0,584 *
0,288
-0,102
0,317
-0,188
0,328
Ongeschoold
-0,092
0,147
-0,245
0,152
-0,215
0,153
Godsdienstige denominatie
Beide katholiek (ref.)
0,000
/
0,000
/
Beide liberaal-protestants
0,262
0,173
0,497 **
0,178
0,432 *
0,190
Beide orthodox-protestants
-0,030
0,269
0,090
0,279
0,150
0,282
Hij katholiek, zij protestants
0,774 **
0,294
0,917 **
0,309
0,750 *
0,317 ogistische regressiecoëfficiënten en standaardfouten van vijf multilevelmodellen van kinderloosheid bij echtparen na 15 jaar huwelijk
Model I
Model II
Model III
Model IV
Model V
b
s.e. b
s.e. b
s.e. b
s.e. b
s.e. Resultaten Orthodox-protes-
tantse koppels verschillen niet of nauwelijks van de referentiegroep. 71 Over de tijd heen blijkt de eerder gesignaleerde trend naar méér kinderloos-
heid statistisch significant te zijn. Er werd ook geëxperimenteerd met een inde-
ling van het interbellum in een fase vóór en een fase na 1933 (niet gerapporteerd
in de tabel), maar de trend naar meer kinderloosheid bleek niet sneller te gaan
tijdens de tweede periode. Het effect van de leeftijd van de man is randje signi-
ficant: niet alleen een hogere huwelijksleeftijd van de vrouw, maar in mindere
en meer onzekere mate ook een hoge huwelijksleeftijd van de man hangt samen
met een hogere kans op kinderloosheid. Overigens werd ook het effect van het
leeftijdsverschil tussen man en vrouw op diverse manieren getest (niet gerap-
porteerd in de tabel), maar dat bleek geen aantoonbare rol te spelen. Ook het
ontbreken van stukjes informatie uit de levensloop van de onderzochte huwelij-
ken hangt niet samen met de kans dat geen enkel overlevend kind geobserveerd
werd. Dit werd op verschillende manieren onderzocht. In de modellen gerappor-
teerd in tabel 4 gebeurt dat aan de hand van opname van het aantal maanden
waarvoor informatie ontbreekt. De variatie in kinderloosheid op het niveau van de 42 onderscheiden regio’s
neemt met ongeveer eenderde af wanneer de huwelijksleeftijd in rekening wordt
gebracht. De geschatte standaarddeviatie van de log-of-odds bedraagt immers
0,300 in model I zonder huwelijksleeftijden, en slechts 0,198 in model II mét
huwelijksleeftijd. Regionale verschillen in kinderloosheid hangen dus, zoals ver-
wacht, samen met regionale verschillen in huwelijkstiming. Model III onderzoekt de samenhang van kinderloosheid met de twee regio-
nale kenmerken, rekening houdend met het huwelijksjaar en de huwelijksleef-
tijd. De schattingen wijzen uit dat kinderloosheid niet of nauwelijks samenhing
met het regionale percentage werklozen in de mannelijke beroepsbevolking in hoge kinderloosheid tijdens het interbellum in nederland Tabel 4 Logistische regressiecoëfficiënten en standaardfouten van vijf multilevelmodellen van kinderloosheid bij echtparen na 15 jaar huwelijk
Model I
Model II
Model III
Model IV
Model V
b
s.e. b
s.e. b
s.e. b
s.e. b
s.e. Resultaten -50,490 ** 19,230
-46,330 *
19,860
-58,790 **
18,800
-58,170 ** 19,000
-49,700 *
20,030
ar
0,025 *
0,010
0,021 *
0,010
0,027 **
0,010
0,027 **
0,010
0,023 *
0,010
eftijd vrouw
0,120 ***
0,016
0,111 *** 0,015
0,109 ***
0,015
0,118 ***
0,016
eftijd man
0,025 ~
0,013
0,026 *
0,013
0,028 *
0,013
0,026
0,014
brekende maanden
0,001
0,001
0,000
0,001
0,001 ~
0,001
0,001
0,001
0,000
0,001
n
oolde arbeider (ref.)
0,000
/
0,000
/
0,000
/
onals & managers
0,454
0,357
0,192
0,368
0,011
0,382
oorden
0,660 ***
0,180
0,502 **
0,186
0,418 *
0,191
olde arbeiders
0,414 *
0,167
0,346 *
0,173
0,269
0,177
en vissers
0,497 *
0,209
0,189
0,219
0,333
0,223
roepsvermelding
2,146 ***
0,260
2,116 ***
0,269
2,054 ***
0,274
melding vrouw
roepsvermelding (ref.)
0,000
/
0,000
/
0,000
/
old
0,584 *
0,288
-0,102
0,317
-0,188
0,328
oold
-0,092
0,147
-0,245
0,152
-0,215
0,153
ige denominatie
tholiek (ref.)
0,000
/
0,000
/
eraal-protestants
0,262
0,173
0,497 **
0,178
0,432 *
0,190
thodox-protestants
-0,030
0,269
0,090
0,279
0,150
0,282
oliek, zij protestants
0,774 **
0,294
0,917 **
0,309
0,750 *
0,317 levelmodellen van kinderloosheid bij echtparen na 15 jaar huwelijk 72 Hij protestants, zij katholiek
0,447
0,286
0,770 **
0,294
0,546
0,307
Liberaal met orthodox-prot. 0,583 **
0,202
0,714 ***
0,210
0,637 **
0,219
Zonder denominatie met kath. 0,557 *
0,258
0,619 *
0,270
0,634 *
0,271
Zonder denominatie met prot. 0,617 **
0,189
0,839 ***
0,195
0,804 ***
0,205
Joods
0,445
0,498
0,660
0,506
0,421
0,514
Beide zonder godsdienst
0,301
0,388
0,589
0,398
0,548
0,404
Regionale kenmerken
% Werkloze mannen 1933
0,006
0,014
-0,003
0,016
-0,006
0,018
% Progressieve stemmen 1937
0,014 *
0,006
0,008
0,006
-0,003
0,007
Gemeentelijke kenmerken
Factor nutsvoorzieningen
0,207 *
0,089
0,075
0,105
Factor aanbod van winkels
0,153 *
0,059
0,189 **
0,063
Factor bezit van motorvoertuigen
-0,041
0,078
-0,060
0,081
Bevolkingsomvang in duizendtallen in 1924
0,0009 ~
0,0005
Random effects
Onverklaarde variatie op
regionaal niveau (Std.)
0,300
0,198
0,198
0,201
0,201
N huwelijken
2.845
2.845
2.845
2.779
2.779
N regio’s
42
42
42
42
42
Deviance
2.304
2.166
2.267
2.212
2.116
~ = p < 0,10; * = p < 0,05; ** = p < 0,01; *** = p < 0,001. Bronnen: HSN datasets Levenslopen en Akten, release 2007.01 en Historische Databank Nederlandse Gemeenten. Resultaten 73 0,447
0,286
0,770 **
0,294
0,583 **
0,202
0,714 ***
0,210
0,557 *
0,258
0,619 *
0,270
0,617 **
0,189
0,839 ***
0,195
0,445
0,498
0,660
0,506
0,301
0,388
0,589
0,398 hoge kinderloosheid tijdens het interbellum in nederland 1933, maar wel met het percentage stemmen voor progressieve partijen bij de
Tweede Kamerverkiezingen van 1937: de kans op geen overlevende kinderen in
verhouding tot de kans op wél overlevende kinderen na vijftien jaar huwelijk
stijgt naar schatting met 15 procent als het percentage stemmen voor progressie-
ve partijen met tien procentpunten toeneemt (exp(10 x 0,014) = 1,15). Deze resul-
taten suggereren dat het culturele klimaat in de regio waar echtparen gingen
wonen van groter belang was dan de regionale intensiteit van de economische
crisis van begin jaren dertig, althans in de mate dat die samenhing met manne-
lijke werkloosheid. 74 74 Model IV voegt de gemeentelijke indicatoren voor het consumptieniveau en
de commerciële voorzieningen toe. De kans op kinderloosheid blijkt groter te
zijn in gemeenten waar een groot deel van de bevolking een radiotoestel in huis
heeft en aangesloten is op gas, elektriciteit en telefoon. Ook in gemeenten met
een groot aanbod van winkels (voeding, kleding, huishoudelijke artikelen) blij-
ken meer echtparen kinderloos te zijn na vijftien jaar huwelijk. Het bezit van
motorvoertuigen blijkt niet of nauwelijks samen te hangen met kinderloosheid. g
j
j
g
Na toevoeging van de gemeentelijke kenmerken verzwakt het effect van het
regionale percentage progressieve stemmen zozeer dat we een nuleffect niet
meer kunnen uitsluiten. Werkloosheid heeft nog steeds geen significant effect
en de richting van het effect draait zelfs om. Dit alles doet vermoeden dat de
gemeentelijke indicatoren voor consumptieniveau en de regionale werkloosheid
en verkiezingsresultaten correleren. Dat blijkt inderdaad het geval. De hoogste
correlatie is die tussen de factor nutsvoorzieningen en het regionale percenta-
ge werklozen (Pearson r = 0,56) en diezelfde factor correleert ook sterk positief
met het percentage progressieve stemmen (r = 0,53). Overigens hangt de factor
nutsvoorzieningen zeer sterk samen met het al dan niet urbane karakter van
de gemeente in kwestie (r = 0,81). De dichotomie urbaan/niet urbaan is echter
niet in de analyses opgenomen, omdat deze variabele minder goed aansluit bij
de theorie dan de variabelen die we wél gebruikten en bovendien minder rijk
aan informatie is. Uit hier niet gerapporteerde analyses bleek inderdaad dat
de dichotome variabele urbaan/niet urbaan geen significant effect heeft op de
kinderloosheid, of de andere gemeentelijke en regionale kenmerken nu worden
opgenomen of niet. De hoge correlatie tussen urbaniteit en nutsvoorzieningen
bleef echter voor scepticisme zorgen over de precieze betekenis van het effect
van de laatstgenoemde factor. hoge kinderloosheid tijdens het interbellum in nederland Mogelijk is de dichotomie urbaan/niet-urbaan een
te ruwe maat. Daarom werd in laatste instantie de bevolkingsomvang van de
gemeenten in het jaar 1924 aan het model toegevoegd: mogelijk zijn er ook per
capita meer mensen op de nutsvoorzieningen aangesloten in grote gemeenten
om de eenvoudige reden dat er in grote gemeenten een groter aanbod van derge-
lijke voorzieningen is? Model V bevat alle besproken verklarende variabelen op zowel individueel,
regionaal als gemeentelijk niveau, plus de gemeentelijke bevolkingsomvang in
1924. Het effect van de nutsvoorzieningen blijkt in grote mate een schijneffect jan van bavel, jan kok en theo engelen te zijn geweest, want het geschatte directe effect verschrompelt in grote mate
na het in rekening brengen van de bevolkingsomvang (van 0,207 tot 0,075, niet
meer statistisch significant). Het effect van het aanbod van winkels, daarente-
gen, verstevigt nog: in gemeenten met een groot aanbod van allerhande winkels
is de kans op kinderloosheid groter, ook na controle voor de bevolkingsomvang
van de gemeente. Het directe effect van de bevolkingsomvang zelf, na controle
voor onder andere de nutsvoorzieningen en het winkelaanbod, is klein: een ver-
gelijking van een gemeente met 1.000 inwoners met één van 10.000 inwoners (de
overige gemeentelijke, regionale en individuele kenmerken constant houdend)
levert een verschil in voorspelde kans op kinderloosheid op van hooguit 0.002. Dit kan statistisch enkel als significant beschouwd worden als een kans op type-
I-fout van 10 procent aanvaard wordt. 75 De eerder besproken verschillen tussen de mannelijke beroepsgroepen
blijven in grote lijnen overeind, met name de relatief hoge kinderloosheid bij
mannen zonder beroepsvermelding en, in tweede orde, bij de witte boorden. Hetzelfde geldt voor de uitgesproken verschillen naar gelang van godsdiensti-
ge gezindte: ook na het in rekening brengen van de regionale en gemeentelijke
kenmerken zijn het de eerder liberaal denkende protestanten en de gemengde
huwelijken die een verhoogde kans op kinderloosheid hebben (zie figuur 2). Conclusies en discussie In het beeld van de Tweede Demografische Transitie, die zich volgens velen
vanaf de jaren zestig aan het voltrekken is, staat de sterk toegenomen vrije keuze
over seksualiteit, huwelijk en kinderen krijgen centraal. In de periode daarvóór
zou de autonomie van stellen ondergeschikt zijn geweest aan groepsnormen,
niet in het minst die van behoudende kerkgenootschappen. In deze optiek wordt
de opmerkelijke toename van kinderloosheid vóór de Tweede Wereldoorlog ver-
klaard uit economische depressie en oorlogsdreiging en niet uit een ontwikke-
ling naar ‘vrije keuze’. Toch zagen contemporaine waarnemers dat wél zo. Zij
wezen op de veranderingen in de voorkeuren van mensen. Steeds belangrijker
werden de zelfontplooiing van vrouwen in een betaalde baan, het uitgaansle-
ven, het behoud van seksuele aantrekkingskracht en – last but not least – het
genieten van een hoog welstandsniveau. Voor een toenemend aantal mensen
betekende dit dat na het huwelijk nog een tijdlang gewacht werd met kinderen
krijgen, wat voor sommigen neerkwam op – al dan niet gepland – afstel. Twee
visies staan tegenover elkaar: de ene ziet kinderloosheid in deze periode als pri-
mair voortkomend uit een keuze voor een vrijere levensstijl, de ander als afge-
dwongen door de economische neergang en politieke onrust. 76 Vrije levensstijl of crisis? Die vraag staat centraal in ons empirisch onder-
zoek, waarin we hebben geprobeerd te kijken welke visie de achtergronden
van kinderloosheid in het interbellum het best benadert. Hiervoor hebben we
gebruikgemaakt van individuele gegevens over 2845 huwelijken uit de HSN, als
ook van contextuele kenmerken van gemeenten en regio’s uit de Historische
Databank Nederlandse Gemeenten. Onze modellen laten zien dat huwelijksuit-
stel een belangrijke rol speelt in de verklaring van kinderloosheid. In een aantal
sociale groepen (met name de midden- en hogere groepen) was het gebruikelijk
te wachten met trouwen en kinderen krijgen tot een acceptabel welstandsniveau
was bereikt. Voor de meeste groepen viel overigens trouwen en kinderen krijgen
samen, immers zodra we gaan controleren voor de huwelijksleeftijd wordt de
associatie met kinderloosheid zwakker. Alleen bij de meest geschoolde groepen
én bij de baanlozen blijkt een relatief grote neiging om ook ná het huwelijk tot
uitstel en afstel van kinderen over te gaan. Interessant is dat controle voor de huwelijksleeftijd het effect van religie
juist sterker doet uitkomen. Hier blijkt heel duidelijk het onderscheid tussen
groepen voor wie het uit- of afstellen van kinderen ‘denkbaar’ was en voor wie
dat niet was. hoge kinderloosheid tijdens het interbellum in nederland Figuur 2 Volgens model V voorspelde kans op kinderloosheid voor modale echtparen in een
gemiddelde gemeente en getrouwd in 1930, naar gelang van godsdienstige gezindtea
0,00
0,05
0,10
0,15
0,20
Beide katholiek
Beide orthodox protestants
Beide liberaal protestants
Joods
Beide zonder
Hij protestant, zij katholiek
Liberaal met orthodox protestants
Zonder denominatie met katholiek
Hij katholiek, zij protestants
Zonder denominatie met protestants
Kans op kinderloosheid Figuur 2 Volgens model V voorspelde kans op kinderloosheid voor modale echtparen in een
gemiddelde gemeente en getrouwd in 1930, naar gelang van godsdienstige gezind Figuur 2 Volgens model V voorspelde kans op kinderloosheid voor modale echtparen in een
gemiddelde gemeente en getrouwd in 1930, naar gelang van godsdienstige gezindtea
0,00
0,05
0,10
0,15
0,20
Beide katholiek
Beide orthodox protestants
Beide liberaal protestants
Joods
Beide zonder
Hij protestant, zij katholiek
Liberaal met orthodox protestants
Zonder denominatie met katholiek
Hij katholiek, zij protestants
Zonder denominatie met protestants
Kans op kinderloosheid Kans op kinderloosheid a Voorspelde waarden voor geschoolde arbeiders met vrouwen zonder beroepsvermelding, alle ande-
re covariaten op hun gemiddelde waarden (huwelijksleeftijd van vrouw en man respectievelijk 25 en a Voorspelde waarden voor geschoolde arbeiders met vrouwen zonder beroepsvermelding, alle ande-
re covariaten op hun gemiddelde waarden (huwelijksleeftijd van vrouw en man respectievelijk 25 en
27 jaar, zie verder tabel 2). B
HSN d
L
l
Ak
l
2007 01
Hi
i h
D
b
k N d l
d a Voorspelde waarden voor geschoolde arbeiders met vrouwen zonder beroepsvermelding, alle ande-
re covariaten op hun gemiddelde waarden (huwelijksleeftijd van vrouw en man respectievelijk 25 en
27 jaar zie verder tabel 2) re covariaten op hun gemiddelde waarden (huwelijksleeftijd van vrouw en man respectievelijk 25 en
27 jaar, zie verder tabel 2). Bronnen: HSN-datasets Levenslopen en Akten, release 2007.01 en Historische Databank Nederlandse
Gemeenten. hoge kinderloosheid tijdens het interbellum in nederland hoge kinderloosheid tijdens het interbellum in nederland Noten 1. Met vruchtbaarheid wordt het aantal geboorten per vrouw bedoeld. Het kindertal, of het
aantal overlevende kinderen, is afhankelijk van de vruchtbaarheid en van de overlevings-
kansen van kinderen. De hoge kinderloosheid bij huwelijken uit het interbellum was een
gevolg van de stijging van de onvruchtbaarheid, want de zuigelingen- en kindersterfte
daalde tijdens deze periode. 1. Met vruchtbaarheid wordt het aantal geboorten per vrouw bedoeld. Het kindertal, of het
aantal overlevende kinderen, is afhankelijk van de vruchtbaarheid en van de overlevings-
kansen van kinderen. De hoge kinderloosheid bij huwelijken uit het interbellum was een
gevolg van de stijging van de onvruchtbaarheid, want de zuigelingen- en kindersterfte
daalde tijdens deze periode. 2. Het betreft (in de terminologie van de volkstelling): Rotterdam, Nijmegen, Restgroep ver-
stedelijkt platteland Noord-Holland, Plattelandsgemeenten Friesland, Plattelandsgemeen-
ten Overijssel en Plattelandsgemeenten Utrecht. 2. Het betreft (in de terminologie van de volkstelling): Rotterdam, Nijmegen, Restgroep ver-
stedelijkt platteland Noord-Holland, Plattelandsgemeenten Friesland, Plattelandsgemeen-
ten Overijssel en Plattelandsgemeenten Utrecht. 3. In verband met de noodzaak exacte huwelijks- en geboortedata te gebruiken, is gebruikge-
maakt van de niet-geanonimiseerde versie, die alleen bij de HSN (Internationaal Instituut
voor Sociale Geschiedenis) geraadpleegd kan worden. 3. In verband met de noodzaak exacte huwelijks- en geboortedata te gebruiken, is gebruikge-
maakt van de niet-geanonimiseerde versie, die alleen bij de HSN (Internationaal Instituut
voor Sociale Geschiedenis) geraadpleegd kan worden. 4. De techniek is geïmplementeerd in de lmer-functie van het open source statistische pak-
ket R, meer bepaald van de lme4-library (Bates & Sarkar, 2006; R development Core Team,
2006). Conclusies en discussie Voor orthodox-protestanten en rooms-katholieken was kinderloos-
heid eigenlijk geen optie, terwijl mensen die hun religie vaarwel hadden gezegd,
of er geen bezwaar tegen hadden iemand met een ander of geen geloof te huwen,
dat wel in hun overwegingen konden betrekken. Het wonen in een stedelijke
omgeving gaf nog een extra impuls om uit- en afstel van kinderen als optie te
zien. Hetzelfde geldt voor het wonen in een gemeente met een groot aanbod van
winkels. Dit suggereert dat men de kinderloosheidsoptie inderdaad eerder koos
als men gewend was geraakt aan een hoog welstandsniveau, zoals de contempo- jan van bavel, jan kok en theo engelen raine waarnemers suggereerden. Daarentegen had het wonen in een gebied met
veel werkloosheid geen effect op het al dan niet kinderen krijgen. Betekent dit dat we de vraag ‘keuze of crisis?’ ten gunste van de eerste kun-
nen beantwoorden? En betekent dit dat de Tweede Demografische Transitie in
de tijd een flink stuk naar voren geschoven mag worden? De tegenstelling is mis-
schien te simplistisch. Voor alle mensen zal de crisis in meer of mindere mate
voelbaar zijn geweest. Als men al niet zelf met werkloosheid werd geconfron-
teerd, dan toch met onzekerheid over de toekomst. We kunnen ons drie groepen
met verschillende reacties voorstellen. Eén groep werd dermate hard door de
crisis getroffen, dat uitstel van (nog meer) kinderen de enige uitweg was. Deze
groep vinden we terug als de baanlozen met een hoge kans op kinderloosheid. Een tweede, grote, groep ondervond veel last van de crisis, maar zal voor andere
oplossingen hebben gekozen dan kinderuitstel. Voor deze groep waren kinde-
ren essentieel voor het gezinsbedrijf (boeren, middenstanders) en/of ze werden
beleefd als een goddelijk geschenk waar geen eigen keuze aan te pas kwam. Een
derde groep hechtte veel aan een hoge levensstandaard, die vaak na lange stu-
die en spaarzaamheid was bereikt. Als hun geloofsovertuiging (of het ontbreken
daarvan) het toeliet, werd de verdediging van die levensstandaard door met kin-
deren krijgen te wachten een denkbare optie. Met andere woorden: de crisis ver-
taalde zich vooral dan in kinderloosheid wanneer kinderen krijgen als een vrije
keuze werd ervaren. 77 Agresti, A. (2002). Categorical data analysis. Hoboken (N.J.), USA: Wiley.
Anderson, M. (1998). Highly restricted fertility: Very small families in the British
fertility decline. Population Studies, 52, 177-199. Literatuur Agresti, A. (2002). Categorical data analysis. Hoboken (N.J.), USA: Wiley. Anderson, M. (1998). Highly restricted fertility: Very small families in the British
fertility decline. Population Studies, 52, 177-199. Anderson, M. (1998). Highly restricted fertility: Very small families in the British
fertility decline. Population Studies, 52, 177-199. Ariès, Ph. (1980). Two successive motivations for the declining birth rate in the
West. Population and Development Review, 6, 645-650. Ariès, Ph. (1980). Two successive motivations for the declining birth rate in the
West. Population and Development Review, 6, 645-650. hoge kinderloosheid tijdens het interbellum in nederland hoge kinderloosheid tijdens het interbellum in nederland hoge kinderloosheid tijdens het interbellum in nederland Bates, D. & D. Sarkar (2006). lme4: linear mixed-effects models using S4 classes. R pac-
kage version 0.995-2. Beekink, E., O. Boonstra, T. Engelen & H. Knippenberg (red.) (2003). Nederland in
verandering. Maatschappelijke ontwikkelingen in kaart gebracht 1800-2000. Amster-
dam: Aksant. 78 Buyst, E. (2006). Van industriële grootmacht tot de ‘zieke man’ van West-Europa. In M. Van den Wijngaert (red.), België, een land in crisis 1913-1950 (pp. 121-173). Antwerpen: Standaard. Carr-Saunders, A. M. (1936). World population. Past growth and present trends. Oxford:
Clarendon Press. Charles, E. (1934). The twilight of parenthood. London: Watts & Co. Coale, A. J. (1986). The decline of fertility in Europe since the eighteenth cen-
tury as a chapter in demographic history. In A. J. Coale & S. C. Watkins (red.),
The Decline of Fertility in Europe (pp. 1-30). Princeton (N.J.): Princeton University
Press. Coontz, S. (2005). Marriage, a history: from obedience to intimacy, or how love conquered
marriage. New York: Viking. Doblhammer, G. (2000). Reproductive history and mortality later in life: a compa-
rative study of England and Wales and Austria. Population Studies, 54, 169-176. Dykstra, P. (ter perse). Childless old age. In P. Uhlenberg (red.), Handbook of Popula-
tion Aging. Berlin: Springer. Dykstra, P.A. & G.O. Hagestad (2007). Roads less taken: developing a nuanced view
of older adults without children. Journal of Family Issues, 28, 1275-1310. Engelen Th., H. Hillebrand & F. van Poppel (1989). Kindertal naar kenmerken, 1900-
1960. ’s- Gravenhage: NIDI. Festy, P. (1979). La fécondité des pays occidentaux de 1870 à 1970. Paris: INED/Presses
Universitaires de France. Frejka, T. & J.-P. Sardon (2004). Childbearing trends and prospects in low-fertility coun-
tries. A cohort analysis (European Studies of Population No. 13). Dordrecht: Klu-
wer. Hakim, C. (2000). Work-Lifestyle Choices in the 21st Century: Preference Theory. Kaa, D.J. van de (2004). Demographic revolutions or transitions? A foreword. In Literatuur Oxford:
Oxford University Press. Hofstee, E.W. (1981). Korte demografische geschiedenis van Nederland van 1800 tot
heden. Haarlem: Fibula-Van Dishoeck. Hooft, F.W. ’t (1926). De maximum-bevolking van Nederland. De Economist, 75,
690-707. Hooft, F.W. ’t (1936). De bevolking van Nederland in crisis. De Economist, 85,
99-111. Hurt, L.S., C. Ronsmans, O.M.R. Campbell, S. Saha, M. Kenward & M. Quigley
(2004). Long-term effects of reproductive history on all-cause mortality among
adults in rural Bangladesh. Studies in Family Planning, 35, 189-196. Jacobson, P.H. (1950). Differentials in divorce by duration of marriage and size of
family. American Sociological Review, 15, 235-244. Kaa, D.J. van de (2004). Demographic revolutions or transitions? A foreword. In T. jan van bavel, jan kok en theo engelen Frejka & J.-P. Sardon, Childbearing trends and prospects in low-fertility countries. A
cohort analysis (pp. xi-xiv). Dordrecht/Boston/London: Kluwer. Frejka & J.-P. Sardon, Childbearing trends and prospects in low-fertility countries. A
cohort analysis (pp. xi-xiv). Dordrecht/Boston/London: Kluwer. Kim, J.-O. & C.W. Mueller (1978). Factor analysis. Statistical methods and practical
issues. London: Sage. 79 Kirk, D. (1946). Europe’s population in the interwar years. Geneva: League of Natio Knippenberg, H. (1992). De religieuze kaart van Nederland. Omvang en geografische
spreiding van de godsdienstige gezindten vanaf de Reformatie tot heden. Assen/Maas-
tricht: Van Gorcum. Kok, J & J. Van Bavel (2006). Stemming the tide. Denomination and religiousness
in the Dutch fertility transition (1845-1945). In R. Derosas & F. van Poppel (red.),
Culture and the Decline of Fertility (pp. 83-105). Dordrecht: Springer Verlag. Kuczynski, R.R. (1932). Fertility and reproduction. Methods of measuring the balance of
births and deaths. New York (N.Y.): Falcon Press. Kuczynski, R.R. (1935). The decrease of fertility. Economica, 2, 128-141. Kumle, M. & E. Lund (2000). Patterns of childbearing and mortality in Norwegian
women. A 20 year follow-up of women aged 40-96 in the 1970 Norwegian cen-
sus. In J.-M. Robine, T. Kirkwood & M. Allard (red.), Sex and longevity: sexuality,
gender, reproduction, parenthood (pp. 117-128). New York: Springer. Landry, A. (1933). La révolution démographique. In Economic essays in honour of
Gustav Cassel (pp. 357-367). London: George Allen & Unwin. Landry, A. (1934). La révolution démographique: études et essais sur les problemes de la
population. Paris: Sirey. Leeuwen, M.H.D. van, I. Maas & A. Miles (2002). HISCO. Historical International
Standard Classification of Occupations. Leuven: Leuven University Press. Leeuwen, M.H.D. van, I. Maas en A. Miles (2005). Literatuur Marriage Choices and Class Bounda-
ries: Social Endogamy in History. Cambridge: Cambridge University Press. Lesthaeghe, R. (1995). The second demographic transition in Western countries:
an interpretation. In K.O. Mason & A.-M. Jensen (red.), Gender and family change
in industrialized countries (pp. 17-62). Oxford: Clarendon Press. Lesthaeghe, R. & D.J. van de Kaa (1986). Twee demografische transities? In R. Lest-
haeghe & D.J. van de Kaa (red.), Bevolking: groei en krimp (pp. 9-24). Boekafleve-
ring bij Mens & Maatschappij jaargang 61. Deventer: Van Loghum Slaterus. Lesthaeghe, R. & J. Surkyn (2004). When history moves on: the foundations and diffu-
sion of a second demographic transition. Brussel: VUB Interface Demography. Lesthaeghe, R. & J. Surkyn (2006). Grondslagen en verspreiding van de Tweede
Demografische Transitie. Justitiële Verkenningen, 32, 75-102. Mandemakers, K. (2000). The Netherlands. Historical Sample of the Netherlands. In P.K. Hall, R. McCaa & G. Thorvaldsen (red.), Handbook of International His-
torical Microdata for Population Research (pp. 149-177). Minneapolis: Minnesota
Population Center. Manor, O., Z. Eisenbach, A. Israeli & Y. Friedlander (2000). Mortality differentials
among women: the Israel longitudinal mortality study. Social Science & Medi-
cine, 51, 1175-1188. hoge kinderloosheid tijdens het interbellum in nederland Methorst, H.W. (1914). De beteekenis van den achteruitgang van het geboortecijfer in
Nederland (Kleine geschriften uitgegeven door de Maatschappij tot Nut van ’t
Algemeen 109). Amsterdam: Van Looy. 80 Methorst, H.W. & M.J. Sirks (1948). Het bevolkingsvraagstuk. Amsterdam: Scheltema
en Holtema. Mombert, P. (1929). Bevölkerungslehre. Jena: Fischer. Mombert, P. (1929). Bevölkerungslehre. Jena: Fischer. Morgan, S.Ph. (1991). Late nineteenth- and early twentieth-century childlessness. American Journal of Sociology, 97, 779-807. Myrdal, A. (1941). Nation and family. The Swedish experiment in democratic family and
population policy. New York/London: Harper & Brothers. NIDI (2003). Bevolkingsatlas van Nederland. Demografische ontwikkelingen van 1850 tot
heden. Rijswijk/Den Haag: Elmar/NIDI. Notestein, F.W. (1950). The population of the world in the year 2000. Journal of the
American Statistical Association, 45, 335-345. Poppel, F. van (1983). Differential fertility in the Netherlands: an overview of long-term
trends with special reference to the post-World War I marriage cohorts. Voorburg:
NIDI. Poppel, F. van (1992). Trouwen in Nederland. Een historisch-demografische studie van de
19e en vroeg 20e eeuw. Den Haag: NIDI. Poston, D.L. & K. Trent (1982). International variability in childlessness. A descrip-
tive and analytical study. Journal of Family Issues, 3, 473-491. Praag, Ph. van (1976). Het bevolkingsvraagstuk in Nederland. Ontwikkelingen van stand-
punten en opvattingen (1918-1940). Literatuur Deventer: Van Loghum Slaterus. Praag, Ph. van (1977). De opkomst van het nieuw-malthusianisme in Vlaanderen. Tijdschrift voor sociale geschiedenis, 3, 192-220. R development Core Team (2006). R: A language and environment for statistical com-
puting. Vienna, Austria: R Foundation for Statistical Computing (URL http://
www.R-project.org). Rowland, D.T. (2007). Historical trends in childlessness. Journal of Family Issues, 28,
1311-1337. Sauvy, A. (1960). Essai d’une vue d’ensemble. In H. Bergues (red.), La prévention des
naissainces dans la famille. Ses origines dans les temps modernes (pp. 377-391). Paris:
Presses Universitaires de France. Schot, J.W. (2001). Techniek in Nederland in de twintigste eeuw. 4: Huishouden, medische
techniek. Zutphen: Walburg Pers. Snijders, T.A.B. & R.J. Bosker (1999). Multilevel analysis. An introduction to basic and
advanced multilevel modeling. London/Thousand Oaks: Sage. Sobotka, T. (2008). Does persistent low fertility threaten the future of European
populations? In J. Surkyn, P. Deboosere & J. Van Bavel (red.), Demographic chal-
lenges for the 21st century. A state of the art in Demography (pp. 27-89). Brussel:
VUB/Academia Press. Vulsma, R.F. (2002). Burgerlijke stand en bevolkingsregister. Den Haag: Centraal
Bureau voor Genealogie. jan van bavel, jan kok en theo engelen Voorlopers in de echtscheidingsrevolutie
De relatie tussen echtscheiding en sociale klasse in de
negentiende en vroeg-twintigste eeuw Matthijs Kalmijn1 Inleiding Uit recent onderzoek blijkt dat er in Nederland, maar ook in andere landen, een
negatieve relatie bestaat tussen de kans op echtscheiding en de sociale positie die
iemand in de samenleving inneemt. Onderzoek dat de sociale positie afmeet aan
het opleidingsniveau laat bijvoorbeeld zien dat in recente huwelijkscohorten,
lager opgeleiden een hogere kans hebben om te scheiden dan hoger opgeleiden
(De Graaf & Kalmijn, 2006; Dronkers, 2002; South, 2001). Gelijksoortige bevindin-
gen zijn gedaan wanneer wordt gekeken naar de inkomenspositie: hoe lager het
huishoudinkomen, des te groter de kans op scheiding (Hannan & Tuma, 1990;
Kalmijn, Loeve & Manting, 2007). Voor deze bevindingen bestaan verschillende verklaringen. Een van de ver-
klaringen wordt gezocht in de financiële problemen waar lagere klassen mee
te maken hebben. Financiële problemen kunnen leiden tot spanningen in een
relatie en dit kan de kans op scheiding vergroten (Poortman, 2005). Een andere
verklaring wordt soms gezocht in de positieve relatie tussen sociale klasse en
intelligentie of cognitieve vermogens (Dronkers, 2002). Slimmere mensen zou-
den meer overdacht een partner kiezen, beter in staat zijn relatieproblemen op
te lossen, en minder impulsief zijn bij het nemen van belangrijke beslissingen in
de levensloop (Herrnstein & Murray, 1994). Een laatste argument voor de negatie-
ve relatie tussen sociale klasse en echtscheiding ligt in gezondheids- en gedrags-
problemen. Uit de echtscheidingsliteratuur is bekend dat mensen met fysieke of
mentale gezondheidsproblemen een grotere kans hebben om te scheiden (Wal-
dron, Hughes & Brooks, 1996). Ander onderzoek laat zien dat verslavingsproble-
men, met name alcohol en drugsgebruik, een determinant zijn van scheiding
(Prescott & Kendler, 2001). Omdat er ook aanwijzingen zijn dat gezondheids- en
verslavingsproblemen meer voorkomen in lagere sociale klassen (Ross & Wu,
1995), zou dit eveneens een verklaring kunnen bieden. Hoewel er zowel empirisch als theoretisch dus veel spreekt voor een nega tieve relatie tussen sociale klasse en echtscheiding, is het de vraag of deze relatie
altijd heeft bestaan. Volgens een theorie van de klassieke gezinssocioloog Goode
zou deze negatieve relatie een recent verschijnsel moeten zijn, een typisch ken-
merk van een samenleving met een hoog echtscheidingsniveau. In een samenle-
ving met een laag echtscheidingsniveau zou volgens Goode de relatie andersom
zijn en zouden juist de hogere sociale klassen meer scheiden dan de lagere soci-
ale klassen (Goode, 1962). Inleiding Het onderliggende argument voor deze stelling is een
– overigens nogal impliciet geformuleerde – notie over diffusie van innovatie (zie
De Graaf & Kalmijn, 2006, voor een reconstructie van Goodes theorie). 82 82 Als echtscheiding nog maar weinig voorkomt in een samenleving, zijn er
sterke normatieve, juridische en financiële drempels om te scheiden. Omdat de
hogere sociale klassen over meer hulpbronnen beschikken, zouden zij beter in
staat zijn om over deze drempels heen te stappen. Hogere klassen zouden bijvoor-
beeld meer financiële zekerheid hebben en daarom zou de vrouw gemakkelijker
op zichzelf kunnen wonen. Een scheiding gaat ook met eenmalige financiële
kosten gepaard en die zouden lagere sociale klassen moeilijk kunnen opbren-
gen. Daarnaast zouden hogere klassen beter kunnen omgaan met de juridische
regels en belemmeringen rond scheiding. Belangrijker nog is dat hogere soci-
ale klassen zich minder zouden aantrekken van de normen tegen scheiding. Zij
zouden als een culturele voorhoede voor het eerst traditionele gezinsnormen
kunnen doorbreken. Dit wil niet per se zeggen dat de huwelijken in de lagere
klassen stabieler zijn dan in de hogere klassen. In de lagere sociale klassen zou
bijvoorbeeld verlating vaker kunnen voorkomen dan in hogere sociale klassen. Naarmate scheiding meer voorkomt en de hogere sociale klassen de normen
tegen scheiding hebben doorbroken, worden de normen zwakker en zouden de
middenklassen en lagere klassen dit gedrag gaan overnemen. Als er ten slotte
geen normatieve of andere drempels meer zijn om te scheiden, keert het verband
tussen sociale klasse en scheiding om. In die situatie spelen dan alleen nog de
eerdergenoemde redenen, zoals financiële spanningen, cognitieve vermogens,
en gezondheids- en gedragsproblemen. De diffusietheorie van Goode is plausibel, maar nog weinig onderzocht. Er
zijn tot nog toe drie trendstudies verricht. Een Amerikaanse studie vindt geen
verandering (over perioden tussen 1950 en 1984) in het effect van opleidingsni-
veau op scheiding (Teachman, 2002). Een Zweedse en een Nederlandse studie vin-
den wel de verwachte verandering naar een negatiever wordend effect van het
opleidingsniveau op de echtscheidingskans (De Graaf & Kalmijn, 2006; Hoem,
1997). In Zweden gaat het om de perioden van 1971 tot en met 1990 en in Neder-
land gaat het om de huwelijkscohorten van 1942 tot en met 1999. Opmerkelijk
in het Zweedse onderzoek is echter dat in de oudste cohorten geen sprake is van
een positieve relatie tussen opleidingsniveau en echtscheiding. Het effect wordt
weliswaar negatief, maar het was oorspronkelijk niet positief. Inleiding Ook in het onder-
zoek van De Graaf en Kalmijn is het initiële effect van het opleidingsniveau niet
helemaal duidelijk. Als de ouderlijke status in het model wordt opgenomen, is er matthijs kalmijn in het oudste cohort geen positief effect van het opleidingsniveau. De ouderlijke
status heeft wel een positief effect in dat cohort, maar dit effect is niet veran-
derd over cohorten als gecontroleerd wordt voor veranderingen in het effect van
opleiding. 83 Al met al zijn er dus wel aanwijzingen voor de theorie van Goode, maar
bestaat er nog onduidelijkheid over de vraag of de relatie tussen sociale klasse
en scheiding positief was in eerdere huwelijkscohorten. Dat laatste is wel een
essentieel deel van de diffusietheorie van Goode die immers stelt dat de leden
van de hogere klasse de eersten zijn die de traditionele normen en waarden rond
huwelijk en gezin durven te doorbreken. Een mogelijke reden voor deze onduidelijkheid is dat recente studies mis-
schien ‘te jong’ zijn om de periode voor de grote scheidingsrevolutie goed in
beeld te krijgen. Misschien was er wel degelijk een positieve relatie tussen soci-
ale klasse en echtscheiding, maar moeten we verder terug in de tijd om deze
relatie te kunnen zien. Figuur 1 laat de trend in scheidingen zien. Na een relatief stabiele periode
tussen 1950 en 1965 begon het scheidingscijfer snel te stijgen. Tussen 1965 en
1985 vervijfvoudigde het aantal scheidingen (per 1000 echtparen). Daarna was er
sprake van trendloze fluctuatie. Als we kijken naar wat er eerder gebeurde valt
op dat er tussen 1860 en 1940 ook al sprake was van een toename. Dit kunnen we
enerzijds aflezen aan de stijging van het echtscheidingscijfer vanaf 1900 zoals
blijkt uit figuur 1. Voor de periode daarvoor publiceert het CBS geen cijfers over
aantallen scheidingen maar kunnen we kijken naar het aandeel gescheidenen in
het totaal van de gehuwde en ooit gehuwde bevolking. In figuur 2 is dit berekend
aan de hand van gegevens uit de tienjaarlijkse volkstellingen. Dergelijke stand-
cijfers zijn natuurlijk niet helemaal te vergelijken met cijfers over gebeurtenis-
sen (hertrouwde gescheidenen vallen er bijvoorbeeld buiten), maar ze geven bij
gebrek aan beter wel een indruk van de trend. Figuur 2 laat zien dat de stijging
in echtscheiding vermoedelijk begon rond 1860 en daarna voortduurde. Figuur 1 laat verder zien dat de initiële stijging werd afgesloten aan het eind
van de Tweede Wereldoorlog met een zeer sterke piek. Inleiding Deze piek heeft wellicht
voorkomen dat de geleidelijke stijging van daarvoor doorzette tussen 1950 en
1965. Met een beetje goede wil zou men dus kunnen stellen dat er eerder sprake
is geweest van een langetermijnstijging in de echtscheidingskans die begonnen
is in het midden van de negentiende eeuw en die na 1965 is versneld. Om die
reden is het ook van belang de relatie tussen sociale klasse en echtscheiding in
een eerdere fase van de geschiedenis te onderzoeken. De vraag in dit hoofdstuk
is daarom: was er in de laat-negentiende en vroeg-twintigste eeuw sprake van een posi-
tieve relatie tussen sociale klasse en echtscheiding? Deze vraag zal in dit hoofdstuk
beantwoord worden op basis van gegevens over personen geboren in de periode
1812-1922. Eerder historisch onderzoek naar echtscheiding in Nederland is verricht
door Van Poppel (1992). Van Poppel geeft onder meer een samenvatting van de voorlopers in de echtscheidingsrevolutie Figuur 1 Aantal scheidingen per 1000 echtparen (1900-2005) 0
2
4
6
8
10
12
1900 1905 1910 1915 1920 1925 1930 1935 1940 1945 1950 1955 1960 1965 1970 1975 1980 1985 1990 1995 2000 2005
Jaar 84 0
1900 1905 1910 1915 1920 1925 1930 1935 1940 1945 1950 1955 1960 1965 1970 1975 1980 1985 1990 1995 2000 2005
Jaar 0
1900 1905 1910 1915 1920 1925 1930 1935 1940 1945 1950 1955 1960 1965 1970 1975 1980 1985 1990 1995 2000
Jaar Bron: CBS Statline. Figuur 2 Percentage gescheidenen in de gehuwde en ooit gehuwde bevolking naar geslacht
(1859-1947)
Bron: CBS Statline. 0,0
0,2
0,4
0,6
0,8
1,0
1,2
1,4
1,6
1,8
2,0
1859
1869
1879
1889
1899
1909
1920
1930
1947
Jaar van volkstelling
%
mannen
vrouwen Figuur 2 Percentage gescheidenen in de gehuwde en ooit gehuwde bevolking naar geslacht
(1859-1947) Bron: CBS Statline. matthijs kalmijn maatschappelijke discussie over echtscheiding in de negentiende eeuw. Hieruit
blijkt dat juridische belemmeringen om te scheiden in die tijd vanuit verschil-
lende politieke perspectieven (feminisme, marxisme, liberalisme) werden bekri-
tiseerd. Deze discussie lijkt sterk op de discussie die werd gevoerd ten tijde van
de verdere liberalisering van de wetgeving in de tweede helft van de twintigste
eeuw (Van Poppel & De Beer, 1993). Van Poppel onderzocht huwelijksregisters uit
Den Haag in de periode 1850-1882 om enkele sociale determinanten van echt-
scheiding op het spoor te komen, waaronder ook de rol van sociale klasse. Inleiding Uit
dit onderzoek blijkt dat de scheidingskans inderdaad hoger is in de categorie
‘intellectuelen en beambten’ dan in de categorie ‘losse arbeiders’. De verschillen
zijn echter niet dramatisch. Van Poppel wijst in zijn interpretatie op de eerderge-
noemde financiële en juridische drempels die losse arbeiders moeilijker konden
overbruggen dan intellectuelen en beambten. In lagere milieus zou ‘verlating’
weer meer voorkomen, maar daarover is het cijfermateriaal niet erg hard. 85 Interessant is dat volgens de gegevens van Van Poppel in de grote burgerij
de scheidingskans juist lager was dan in de andere sociale klassen. Dit lijkt in
tegenstelling te zijn tot het idee van Goode. Van Poppel wijst hier op een werking
van normatieve drempels die juist andersom is dan Goode veronderstelde in de
diffusietheorie. In hogere milieus zou het overtreden van traditionele normen
meer sociale sancties opleveren en zou men dus juist niet een culturele voorhoe-
derol kunnen spelen. Opmerkelijk is voorts dat binnen de categorie ‘intellectu-
elen en beambten’ de artistieke beroepen de hoogste scheidingscijfers hadden. Omdat dit ook een groep is met doorgaans meer liberale denkbeelden, zou zij
wel gemakkelijker traditionele normen kunnen doorbreken. Hoewel Van Poppel
dit niet noemt, wijzen zijn bevindingen wellicht op het verschil tussen de eco-
nomische en culturele elite (Bourdieu, 1979; Ganzeboom, 1988; Lamont, 1992). Mogelijk was het niet de elite als geheel die een voortrekkersrol vervulde, maar
was het de culturele elite die dat deed en niet de vaak meer behoudende econo-
mische elite. Data en methode De data zijn afkomst uit de Historische Steekproef Nederlandse bevolking (HSN release
akten 2007.01 en HSN release akten_h_scheiding 2007.01.). Voor meer informatie over
de HSN, zie www.iisg.nl/~hsn. De gegevens die hier gebruikt worden zijn afkom-
stig uit alle beschikbare huwelijksakten die in het kader van het grootschalige
HSN-project momenteel zijn verzameld. Deels horen deze akten bij de steekproef
van geboorteakten in de periode 1850-1922 uit de provincies Friesland, Zeeland,
Utrecht, en de stad Rotterdam, en geboorteakten uit de periode 1883-1922 uit de
rest van het land, waarvoor levensloopgegevens zijn verzameld. Daarnaast zijn er
ook huwelijksakten in het bestand van HSN-personen die buiten de levensloop-
steekproef zijn gebleven. Hierbij gaat het vooral om huwelijken gesloten in de
geboortegemeente. Gezien het geringe aantal echtscheidingen in de hier geana- voorlopers in de echtscheidingsrevolutie lyseerde periode analyseer ik in eerste instantie alle beschikbare huwelijken. In
totaal gaat het dan om 21,763 akten, waarvan 1,190 akten betrekking hebben op
tweede of derde huwelijken. In een additionele analyse kijk ik naar specifieke
subsets van de akten om te zien hoe robuust mijn bevindingen zijn. Meer in
het bijzonder kijk ik naar een subset van akten die behoren bij de levensloop-
gegevens en een subset van akten waarbij huwelijken gesloten zijn voor 1930. Omdat in deze subsets het aantal scheidingen fors daalt, is de statistische zeg-
gingskracht van deze analyses echter beperkter dan in de volledige set akten. 86 86 Het grootste deel van de huwelijken (de middelste 90%) is gesloten in de peri-
ode 1848-1936 en het gemiddelde huwelijksjaar is 1898. De risicojaren waarin we
deze huwelijken kunnen bekijken gaan dus vooraf aan de grote scheidingsrevo-
lutie. Van de huwelijken eindigde 542 huwelijken in een scheiding, ongeveer 2
procent. Echtscheiding was in deze tijd natuurlijk een vrij zeldzaam verschijn-
sel. Echtscheidingen zijn bijgeschreven op de huwelijksakten, zodat de construc-
tie van deze variabele onproblematisch is. De determinanten van echtscheiding worden geanalyseerd met een gebeur-
tenissenanalyse. De gegevens zijn omgezet in een persoonsjarenbestand. Vervol-
gens is op dit persoonsjarenbestand een logistische-regressievergelijking geschat
voor de kans op echtscheiding in een jaar, gegeven dat een persoon nog gehuwd
was in dat jaar. De risicoperiode begint bij het jaar van huwelijk en eindigt bij
scheiding of in het overlijdensjaar (right censoring). Voor ongeveer eenderde van
de onderzoekspersonen in de huwelijksakten is geen sterftedatum bekend. In
de event-history-analyse dienen waarnemingen van een huwelijk gecensureerd te
worden als de onderzoekspersoon overlijdt. Data en methode Om hier toch een beeld van te krij-
gen zijn ontbrekende overlijdensjaren geschat. Dit is gebeurd aan de hand van
een regressievergelijking waarbij de levensduur de afhankelijke variabele is en
geboortejaar, geslacht, analfabetisme, stedelijkheid en regio de onafhankelijke
variabelen zijn. De berekening is als volgt: overlijdensjaar = geboortejaar + [ -53,669 + 1,136*vrouw + 0,067*geboorte-
jaar - 2,681*grote stad - 5,445*analfabetisme + 0,983*zuid + 2,985*west ], waarbij alle variabelen behalve geboortejaar 0/1 variabelen zijn. Alle effecten
zijn statistisch significant. Het geschatte overlijdensjaar is afgerond op hele
getallen. waarbij alle variabelen behalve geboortejaar 0/1 variabelen zijn. Alle effecten
zijn statistisch significant. Het geschatte overlijdensjaar is afgerond op hele
getallen. Een ander probleem is dat er geen informatie beschikbaar is over de over-
lijdensdatum van de partner. Normaal gesproken zou de risicoperiode van een
huwelijk ook beëindigd (‘gecensureerd’) moeten worden wanneer de partner
overlijdt. Vanuit de onderzoekspersoon bezien is verweduwing immers een alter-
natieve manier waarop het huwelijk kan worden ontbonden,een zogeheten com-
peting risk van scheiding. In de huidige analyse wordt de huwelijksperiode gecen-
sureerd wanneer de onderzoekspersoon overlijdt, maar loopt deze door wanneer
de partner overlijdt voordat de onderzoekspersoon overlijdt. Deze beperking kan matthijs kalmijn de schattingen van de effecten in het model vertekenen, maar het is niet duide-
lijk in welke mate of in welke richting. Om dit na te gaan kijk ik tevens naar een
subset van de waarnemingen waarbij de onderzoekspersoon jonger is dan 55. Vóór deze leeftijd kwam verweduwing minder vaak voor, maar komt scheiding
wel veel voor. 87 Variabelen en hun meting De klasse is bepaald aan de hand van het beroep. Omdat er sprake is van ontbre-
kende waarden, zijn verschillende beroepen gebruikt. Het beroep van de brui-
degom op het moment van huwelijk is als uitgangspunt genomen. In 24 pro-
cent van de gevallen ontbreekt een te gebruiken beroepstitel. In die gevallen is
gebruikgemaakt van het beroep van de vader van de bruidegom op het moment
van het huwelijk. Dat reduceert het aantal ontbrekende waarden tot 19%. Daarna
is het beroep van de vader van de onderzoekspersoon bij geboorte en tot slot het
beroep van de vader van de bruid op het moment van huwelijk gebruikt (daling
tot 9%). Het beroep van de bruid is niet gebruikt, omdat in traditionele samen-
levingen het beroep van de bruidegom meer zegt over de sociaal-economische
positie dan het beroep van de bruid. De indeling in klassen is gebaseerd op HISCO (Van Leeuwen, Maas & Miles,
2002). Omdat het aantal scheidingen niet erg groot is, is gebruikgemaakt van
een indikking van HISCLASS in een beperkt aantal klassen (Maas & Van Leeuwen,
2004): – landarbeiders (8,4%), – boeren en vissers (17,1%), – lagere handarbeid (lager geschoold en ongeschoold) (23,4%), – hogere handarbeid (middelgeschoold) (22,4%), – lagere hoofdarbeid (laaggeschoold) (3,8%), – lagere en hogere managers (4,3%), – lagere en hogere managers (4,3%), – hogere en middelbare hoofdarbeid inclusief hogere en lagere professionals
(11,9%). – hogere en middelbare hoofdarbeid inclusief hogere en lagere professionals
(11,9%). Het is tevens interessant om enkele andere mogelijke determinanten van de
scheidingskans in de analyse te betrekken. Na een basismodel met alleen sociale
klasse, duur en huwelijksjaar, voeg ik een set regionale variabelen toe: (a) geboor-
te in een van de tien grootste steden, en (b) geboorteregio, verdeeld in Noord
(Groningen, Friesland, Overijssel, Drenthe), Zuid (Zeeland, Limburg, Noord-Bra-
bant, Gelderland), en West (Noord-Holland, Zuid-Holland, Utrecht). Het is van
belang voor urbanisatie en regio te controleren, omdat deze samenhangen met
de beroepsgroep en tevens een zelfstandig effect kunnen hebben op de schei-
dingskans (De Graaf & Kalmijn, 1999). Met andere woorden, effecten van klasse
kunnen spurieus zijn, veroorzaakt door onderliggende regionale verschillen en
effecten. Variabelen en hun meting voorlopers in de echtscheidingsrevolutie Tabel 1 Gebeurtenissenanalyse van de kans op scheiding: regressiecoëfficiënten uit een logistische
regressievergelijkinga
Model 1
Model 2
Model 3
b
p
b
p
b
p
Duur
0
-1,135 *
(0,02)
-1,135 *
(0,02)
-1,136 *
(0,02)
1-2
-
-
-
3-5
0,314
(0,15)
0,315 *
(0,15)
0,317
(0,15)
6-8
0,516 *
(0,02)
0,519 *
(0,02)
0,523 *
(0,01)
9-11
0,559 *
(0,01)
0,565 *
(0,01)
0,570 *
(0,01)
12-15
0,657 *
(0,00)
0,666 *
(0,00)
0,674 *
(0,00)
16-19
0,110
(0,62)
0,122
(0,58)
0,132
(0,55)
20-29
-0,489 *
(0,02)
-0,471 *
(0,03)
-0,464 *
(0,03)
30+
-2,076 *
(0,00)
-2,063 *
(0,00)
-2,083 *
(0,00)
Huwelijksjaar
0,030 *
(0,00)
0,029 *
(0,00)
0,031 *
(0,00)
Sociale klasse
Boer
-2,142 *
(0,00)
-1,662 *
(0,00)
-1,579 *
(0,00)
Landarbeider
-1,143 *
(0,00)
-0,743 *
(0,01)
-0,814 *
(0,01)
Lagere handarbeid
0,233 *
(0,04)
0,231 *
(0,05)
0,160
(0,17)
Hogere handarbeid (ref.)
-
-
-
Lagere hoofdarbeid
0,345 ~
(0,06)
0,096
(0,60)
0,153
(0,40)
Manager
-0,127
(0,58)
-0,045
(0,85)
-0,006
(0,98)
Hogere hoofdarbeid
0,504 *
(0,00)
0,435 *
(0,00)
0,454 *
(0,00)
Klasse ontbreekt
-0,438 ~
(0,07)
-0,118
(0,62)
-0,212
(0,38)
Grote stad
0,784 *
(0,00)
0,747 *
(0,00)
West (versus noord/oost)
0,391 *
(0,00)
0,357 *
(0,00)
Zuid (versus noord/oost)
-0,420 *
(0,01)
-0,420 *
(0,01)
Huwelijksleeftijd vrouw
-0,087 *
(0,00)
Man ouder dan vrouw
0,033
(0,73)
Vrouw ouder dan man
0,669 *
(0,00)
Tweede huwelijk
1,379 *
(0,00)
Analfabeet
0,475 *
(0,05)
Constante
-65,128 *
(0,00)
-62,678 *
(0,00)
-65,667 *
(0,00)
Chi-2 toets voor klasse
verschillen
97,0 *
56,1 *
54,1 *
Chi-2
904 *
1082 *
1205 *
~ = p < 0,10; * = p < 0,05 (tweezijdige toetsen). 88 88 ~ = p < 0,10; * = p < 0,05 (tweezijdige toetsen). ~ = p < 0,10; * = p < 0,05 (tweezijdige toetsen). a Aantal huwelijken is 21.763, aantal scheidingen is 542, aantal persoonsjaren is 964.796. a Aantal huwelijken is 21.763, aantal scheidingen is 542, aantal persoonsjaren is 964.796. Waarneming gecensureerd bij sterfte van de onderzoekspersoon. Toets op klasseverschillen geba-
d
d t
d
tb
k
d
d
kl Waarneming gecensureerd bij sterfte van de onderzoekspersoon. Toets op klasseverschillen geba-
seerd op data zonder ontbrekende waarden voor klasse Waarneming gecensureerd bij sterfte van de onderzoekspersoon. Toets op klasseverschillen geba op data zonder ontbrekende waarden voor klasse. Variabelen en hun meting seerd op data zonder ontbrekende waarden voor klasse. Bron: HSN release akten 2007.01 en HSN release akten_h_scheiding 2007.01. matthijs kalmijn In een derde model voeg ik mogelijke verklarende variabelen toe, te weten:
(a) de huwelijksleeftijd van de vrouw, (b) het leeftijdsverschil met de man (ver-
deeld in man drie jaar ouder of meer en man drie jaar jonger of meer), (c) het
rangnummer van het huwelijk (tweede en latere huwelijken worden vergeleken
met eerste huwelijken), en (d) analfabetisme (of man en/of vrouw analfabeet is,
zoals blijkend uit het ontbreken van een handtekening op de huwelijksakte). We
kunnen verwachten dat een deel van de invloed van de sociale klasse verklaard
kan worden door deze demografische en sociale kenmerken. 89 Resultaten De resultaten van de gebeurtenissenanalyse zijn vermeld in tabel 1. We kijken
eerst naar de duurafhankelijkheid. Deze is gemodelleerd in de gebeurtenissen-
analyse en tevens in een figuur weergegeven (figuur 3). De figuur laat de geobser-
veerde conditionele kansen zien op een echtscheiding voor elke eenjarige duur
van het huwelijk. Omdat de geobserveerde kansen fluctueren, is tevens de best
passende trendlijn geschat. We zien dat het eerste jaar een zeer lage scheidings-
kans heeft. Daarna neemt het scheidingscijfer gestaag toe en het is vervolgens op
zijn hoogst in jaar twaalf tot vijftien van het huwelijk. Na vijftien jaar zakt het
cijfer weer tot zeer lage niveaus, hetgeen laat zien dat de minder goede huwe-
lijken er steeds meer worden uitgeselecteerd. Op grond van de geobserveerde
kansen kan berekend worden dat na vijftien jaar huwelijk 2 % door scheiding Figuur 3 Conditionele kansen op scheiding naar duur van het huwelijk Figuur 3 Conditionele kansen op scheiding naar duur van het huwelijk Figuur 3 Conditionele kansen op scheiding naar duur van het huwelijk
Bron: HSN-release-akten 2007.01 en HSN release akten_h_scheiding 2007.01. 0,00
0,05
0,10
0,15
0,20
0,25
0,30
0
5
10
15
20
25
30
Duur
In percentages Bron: HSN-release-akten 2007.01 en HSN release akten_h_scheiding 2007.01. voorlopers in de echtscheidingsrevolutie uit elkaar is gegaan. Dit getal is veel lager dan het huidige niveau. Voor cohorten
getrouwd in 1985 is bijna 20 % ontbonden na vijftien jaar, ongeveer tien keer
zoveel (Garssen, De Beer, Cuyvers & De Jong, 2001). 90 90 Het huwelijksjaar heeft een sterk positief effect, hetgeen laat zien dat over de
huwelijkscohorten 1850-1930 de scheidingskans stijgt. De stijging is gemiddeld
3 % per jaar bij een lineaire schatting. Een kwadratische term voor het huwe-
lijksjaar had geen significant effect. Hieruit blijkt dat er in deze gegevens sprake
is van een min of meer rechtlijnige toename in de scheidingskans over huwe-
lijkscohorten. We kunnen de trend ook met een periode-effect in plaats van met
een cohorteffect schatten. Een lineair periode-effect is 3 % per jaar, identiek aan
het cohorteffect. Als we jaren verdelen in perioden zien we ook een duidelijke
toename: 0 vóór 1900, 1,02 in 1900-1919, 1,79 in 1920-1942, 2,44 in 1943-1948 en
2,05 in 1949-1965 (effecten op de log odds en alle effecten significant). De stijging
is lineair, maar na de naoorlogse piek daalt de kans weer. Resultaten Zoals ook in modern onderzoek wordt gevonden, blijkt de scheidings-
kans hoger in de tien grootste steden dan elders. Ook zijn er regionale verschil-
len, met de hoogste scheidingscijfers in het westen en de laagste cijfers in het
zuiden. Na opname van deze factoren veranderen de effecten van sociale klasse. Het verschil tussen de agrarische beroepen en de handarbeiders wordt kleiner,
maar blijft significant. Dit heeft duidelijk te maken met de sterke relatie tussen
stedelijkheid en het aandeel agrarische beroepen in de bevolking. Na opname van geografische factoren neemt ook het verschil tussen de
hogere handarbeid en lagere hoofdarbeid af. Dit verschil is niet meer significant. Het contrast tussen hogere handarbeiders en hogere hoofdarbeiders neemt ech-
ter slechts af met 14 % en blijft sterk en statistisch significant. Hogere hoofdar-
beiders hebben een 1,54 keer zo hoge scheidingskans als hogere handarbeiders
na constant houden van regionale verschillen en verschillen in stedelijkheid. De
klassenverschillen als geheel blijven overigens significant, maar de X2-waarde is
lager. In model 3 voegen we mogelijke intermediërende variabelen toe. In over-
eenstemming met eerder onderzoek blijkt een jonge huwelijksleeftijd samen te
gaan met een verhoogde scheidingskans (Kalmijn & Poortman, 2006). Daarnaast
zien we dat tweede en latere huwelijken eerder stuklopen dan eerste huwelij-
ken. Ook dit is een goed gedocumenteerd verband in de moderne tijd (Booth &
Edwards, 1992); alleen gaat het in de huidige analyses voornamelijk om wedu-
wen en weduwnaars, en niet om eerder gescheiden personen. Tot slot vinden we
dat huwelijken met een groot leeftijdverschil instabieler zijn. Dit geldt echter
alleen als de vrouw een paar jaar ouder is dan de man, niet als de man een paar
jaar ouder is dan de vrouw. In eerder onderzoek werd eveneens een dergelijk
asymmetrisch effect gevonden (Kalmijn & Poortman, 2006). Ook opvallend is de invloed van analfabetisme, zoals afgemeten aan het al
of niet zetten van een handtekening. Paren waarvan ten minste een van de part-
ners niet kan schrijven hebben een 1,61 keer zo hoge scheidingskans als paren
die wel kunnen schrijven. Omdat hier gecontroleerd wordt voor sociale klasse,
wijst dit mogelijk op een vroege invloed van cognitieve vermogens, zoals die ook
bepleit is voor de tegenwoordige tijd door Dronkers (2002). Hier tegenover staat
dat ongeletterden minder goed hun weg konden vinden in het administratieve
systeem waarin men terechtkomt als men wil scheiden (Van Poppel, 1992). Resultaten Zoals we weten stijgt
de echtscheidingskans daarna zeer sterk, zodat de daling na 1948 gekenschetst
kan worden als een stilte voor de storm. Deze trends komen overeen met wat we
weten uit de bevolkingsstatistiek (figuur 1). Het effect van sociale klasse als geheel is statistisch significant, getuige de
X2-toets dat alle klasseneffecten 0 zijn (onder in tabel 1). Dit laat zien dat de
scheidingskansen significant tussen sociale klassen verschillen. Is er sprake van
een positieve relatie met sociale klasse? Om die vraag te beantwoorden kijken we
naar de effecten in model 1. Hogere handarbeiders zijn de referentiegroep, maar
ik zal ook vergelijkingen maken met andere groepen waar dat zinvol is. We zien
in de eerste plaats dat landarbeiders en boeren een zeer veel lagere scheidings-
kans hebben dan hogere handarbeiders. Ook als we hen met lagere handarbei-
ders vergelijken zijn de kansen significant lager. Hierbij hebben landarbeiders
weer een hogere scheidingskans dan boeren (b = 0,999, p = 0,02). Als we hoofdarbeiders en handarbeiders vergelijken zien we ook significan-
te contrasten. Lagere hoofdarbeiders hebben een marginaal significant hogere
scheidingskans dan hogere handarbeiders. De odds zijn een factor 1,41 hoger. Sterker is het contrast tussen hogere handarbeiders en hogere en middelbare
hoofdarbeiders. Hogere hoofdarbeiders hebben 1,66 keer zo hoge odds om te
scheiden dan hogere handarbeiders. Managers zijn een opvallende uitzonde-
ring, zij hebben een vrij lage scheidingskans, vergelijkbaar met die van hogere
handarbeiders. Het contrast tussen managers en hogere hoofdarbeiders (onder
wie professionals) is eveneens significant (b = 0,63, p = 0,01). Dit bevestigt het
idee dat er binnen de elite een contrast bestaat tussen economische en culturele
groepen. In elk geval hebben managers geen verhoogde scheidingskans en vor-
men zij dus een uitzondering op de hypothese van Goode. Binnen de groep handarbeiders zijn er geringe verschillen, alhoewel de lage-
re handarbeiders een iets hogere scheidingskans blijken te hebben dan hogere
handarbeiders (p = 0,04). Dit resultaat was niet verwacht. Hiervoor vergeleken we
steeds de hoofdarbeiders met de hogere handarbeiders. Als we hoofdarbeiders matthijs kalmijn vergelijken met lagere landarbeiders worden de contrasten wat zwakker. De ver-
schillen met hogere hoofdarbeiders blijven echter significant. Hogere hoofdar-
beiders hebben 1,31 keer zo hoge odds om te scheiden als lagere handarbeiders
(p = 0,02). Dit ondersteunt opnieuw de hypothese van een positieve relatie tussen
echtscheiding en sociale klasse. 91 We kijken vervolgens naar model 2 waarin ook regionale factoren zijn opge-
nomen. ~ = p < 0,10; * = p < 0,05 (tweezijdige toetsen). Resultaten Dat
zou juist een negatief effect van analfabetisme op scheiding suggereren. Contro-
leren voor de invloed van demografische factoren en analfabetisme verandert
opvallend genoeg weinig aan de effecten van sociale klasse. voorlopers in de echtscheidingsrevolutie Tot slot gaan we na hoe robuust de bevindingen zijn. In tabel 2 presenteer ik
eerst analyses waarbij de persoonsjaren zijn beperkt tot jaren waarin de onder-
zoekspersoon jonger is dan 55. Op deze leeftijd is verweduwing van de onder-
zoekspersoon nog relatief zeldzaam. De schattingen zijn gebaseerd op model 2
uit tabel 1. De resultaten uit de eerste kolom van tabel 2 komen overeen met
de resultaten uit tabel 1. Hogere hoofdarbeiders hebben een significant hogere
scheidingskans dan hogere handarbeiders, managers wijken niet af van hand-
arbeiders, en boeren en landarbeiders hebben de laagste scheidingskans. Ook
de grootte van de effecten komt goed overeen. Deze bevindingen laten zien dat
het ontbreken van informatie over verweduwing van de onderzoekspersoon niet
leidt tot vertekende effecten van sociale klasse. 92 In de tweede kolom van tabel 2 laat ik meer recente huwelijksjaren uit de
analyse weg. De gegevens zijn hier beperkt tot huwelijken gesloten in de peri-
ode vóór 1930. Ook hier komen de resultaten goed overeen. Het verschil tussen
landarbeiders en hogere handarbeiders wordt iets kleiner, maar het contrast tus-
sen managers en hogere hoofdarbeiders wordt groter. Het verschil tussen hogere
hoofdarbeiders en hogere handarbeiders is nagenoeg identiek aan tabel 1. Tabel 2 Effecten van klasse op scheiding in gebeurtenissenanalyse voor verschillende subsetsa
Gecensureerd bij
leeftijd 55
Huwelijksjaren
< 1930
Huwelijkskakten
behorende bij
levensloopdata
b
p
b
p
b
p
Boer
-1,517 *
(0,00)
-1,725 *
(0,00)
-0,835 *
(0,04)
Landarbeider
-0,721 *
(0,02)
-0,508 ~
(0,10)
-0,644
(0,14)
Lagere handarbeid
0,250 *
(0,04)
0,248 ~
(0,06)
0,380 *
(0,04)
Hogere handarbeid (ref.)
-
-
-
Lagere hoofdarbeid
0,150
(0,43)
-0,016
(0,94)
0,186
(0,50)
Manager
-0,012
(0,96)
-0,272
(0,36)
0,063
(0,86)
Hogere hoofdarbeid
0,454 *
(0,00)
0,451 *
(0,00)
0,710 *
(0,00)
Klasse ontbreekt
-0,102
(0,69)
-0,040
(0,88)
-0,166
(0,71)
Aantal huwelijken
21546
19443
6433
Aantal scheidingen
492
399
238
Chi-2 toets voor
klasseverschillen
49,8 *
42,9 *
28,2 * el 2 Effecten van klasse op scheiding in gebeurtenissenanalyse voor verschillende subsetsa el 2 Effecten van klasse op scheiding in gebeurtenissenanalyse voor verschillende subsetsa a Gecontroleerd voor regio, urbanisatiegraad, huwelijksjaar, en duurafhankelijkheid. Toets op klasseverschillen gebaseerd op data zonder ontbrekende waarden voor klasse. Toets op klasseverschillen gebaseerd op data zonder ontbrekende waarden voor klasse. Resultaten Bron: HSN release akten 2007.01 en HSN release akten_h_scheiding 2007.01. matthijs kalmijn In de derde kolom beperk ik me tot huwelijken die behoren bij de levens-
loopgegevens, dat wil zeggen de steekproef van geboorteakten in de periode
1850-1922 uit de provincies Friesland, Zeeland, Utrecht, en de stad Rotterdam, en
geboorteakten uit de periode 1883-1922 uit de rest van het land. Dit betekent een
fors lagere N, maar de belangrijkste resultaten blijven overeind. Het contrast tus-
sen hogere hoofdarbeiders en hogere handarbeiders wordt wat groter, net als het
contrast tussen hogere hoofdarbeiders en lagere handarbeiders. De contrasten
tussen boeren en landarbeiders enerzijds, en handarbeiders anderzijds worden
zwakker, maar blijven in het geval van boeren significant. De toets op klassenver-
schillen als geheel blijft zelfs bij het kleinere aantal huwelijken en scheidingen
statistisch significant. Al met al lijken de resultaten redelijk robuust. 93 Conclusie en discussie Volgens een klassieke theorie zou echtscheiding, net als allerlei andere sociale
en culturele vernieuwingen, eerst opkomen in de hogere sociale klassen en later
doorsijpelen naar de lagere sociale klassen (Goode, 1962). De normatieve, eco-
nomische en juridische drempels om te scheiden in een tijd dat scheiding nog
ongebruikelijk is zijn immers gemakkelijker te overbruggen door groepen met
meer sociaal, cultureel en economisch kapitaal. Veel onderzoek naar deze theo-
rie is er niet, maar recentelijk is wel aangetoond dat in het midden van de twin-
tigste eeuw de relatie tussen opleiding en scheiding sterk negatief werd. Op dit
moment zijn het de lagere opleidingsgroepen die vaker scheiden dan de hogere
opleidingsgroepen (De Graaf & Kalmijn, 2006). Minder duidelijk is of er vooraf-
gaand aan de grote toename in echtscheiding wel sprake was van een duidelijk
positieve relatie tussen echtscheiding en opleidingsniveau. Contemporaine sur-
veydata gaan daarvoor ook niet ver genoeg terug in de tijd. Het huidige onderzoek bekijkt een eerdere periode – namelijk de huwelijks-
periode 1848-1936 – en gebruikt sociale klasse als indicator. De analyses uit deze
bijdrage versterken de conclusies van De Graaf en Kalmijn. De resultaten laten
zien dat er in deze vroege periode een duidelijke positieve relatie bestond tus-
sen de kans op echtscheiding en de sociale klasse. Landarbeiders hebben een
zeer lage kans op scheiding, handarbeiders een wat hogere, en hoofdarbeiders
en professionals hebben de hoogste kans op scheiding. Er is hier sprake van een
duidelijke klassengradiënt. Managers vormen een uitzondering op dit patroon. Hun scheidingskansen lijken meer op de kansen van handarbeiders dan op de
kansen van hoofdarbeiders. Er is zelfs een significant verschil in echtscheidings-
kans tussen hogere hoofdarbeiders (onder wie veel professionals) en managers,
hetgeen laat zien dat binnen de hogere groepen, het vooral de culturele elite is
die ‘innoveert’ en waarschijnlijk niet de veelal behoudender economische elite. Dit is een nuancering van de klassieke theorie van Goode. Er is één kanttekening te plaatsen bij de analyses. De analyse is gericht op
formele scheiding en niet op het uit elkaar gaan van een huishouden. Juist in voorlopers in de echtscheidingsrevolutie een situatie waarin de drempels om te scheiden hoog zijn, kunnen partners bij
huwelijksproblemen ervoor kiezen apart te gaan wonen zonder hun huwelijk te
ontbinden. Een speciaal geval daarvan is verlating. Hierbij verbreekt iemand de
huwelijksband door permanent te vertrekken uit het huishouden zonder mede-
weten van of overleg met de partner. Noot 1. Met dank aan Kees Mandemakers en Ineke Maas voor opmerkingen en Kees Mandemakers
voor zijn advies bij de data-analyses. 1. Met dank aan Kees Mandemakers en Ineke Maas voor opmerkingen en Kees Mandemakers
voor zijn advies bij de data-analyses. Conclusie en discussie Verlating kwam doorgaans veel voor bij
huwelijksconflicten en vormde een probleem voor de achterblijvende partner
als deze wilde hertrouwen. Verlating is wel onderzocht, onder meer in het begin
en midden van de twintigste eeuw (Kephart & Monahan, 1952; Zuckerman, 1950),
en in eerdere historische perioden (Butler, 2006). De relatie met sociale klasse is
echter niet goed gedocumenteerd, hoewel men het in het dagelijks spraakge-
bruik wel vaak heeft over verlating als the poor man’s divorce. 94 Het is mogelijk dat een lagere scheidingsfrequentie bij de lagere sociale
klassen werd gecompenseerd door een hogere verlatingsfrequentie. Dit zou een
interessante bevinding zijn, maar niet noodzakelijkerwijs in tegenspraak met de
theorie van Goode. Deze theorie gaat immers over de drempels die er bestaan
voor echtscheiding en niet over wat er in een huwelijk speelt. De positieve relatie
tussen sociale klasse en scheiding betekent dan ook niet dat de huwelijken in de
hogere milieus minder goed waren. Het betekent (waarschijnlijk) alleen dat er
in de verschillende milieus andere oplossingen werden gekozen voor hetzelfde
onderliggende probleem. 1. Met dank aan Kees Mandemakers en Ineke Maas voor opmerkingen en Kees Mandemakers
voor zijn advies bij de data-analyses. Literatuur Booth, A. & J.N. Edwards (1992). Starting over: Why remarriages are more unsta-
ble. Journal of Family Issues, 13, 179-194. Booth, A. & J.N. Edwards (1992). Starting over: Why remarriages are more unsta-
ble. Journal of Family Issues, 13, 179-194. Bourdieu, P. (1979). Distinction: A social critique of the judgement of taste. London:
Routledge & Kegan Paul. Bourdieu, P. (1979). Distinction: A social critique of the judgement of taste. London:
Routledge & Kegan Paul. Butler, S. (2006). Runaway wives: Husband desertion in medieval England. Journal
of Social History, 40, 337-359. Butler, S. (2006). Runaway wives: Husband desertion in medieval England. Journal
of Social History, 40, 337-359. Dronkers, J. (2002). Bestaat er een samenhang tussen echtscheiding en intelligen-
tie? Mens & Maatschappij, 77, 25-42. Dronkers, J. (2002). Bestaat er een samenhang tussen echtscheiding en intelligen-
tie? Mens & Maatschappij, 77, 25-42. Ganzeboom, H.B.G. (1988). Leefstijlen in Nederland; een verkennende studie. Rijswijk:
Sociaal en Cultureel Planbureau. Garssen, J., J. de Beer, P. Cuyvers & A. de Jong (2001). Samenleven: Nieuwe feiten over
relaties en gezinnen. Voorburg: CBS. Goode, W.J. (1962). Marital satisfaction and instability: A cross-cultural class ana-
lysis of divorce rates. International Social Science Journal, 14, 507-526. Graaf, P.M. de & M. Kalmijn (1999). Verschillen in echtscheidingscijfers tussen matthijs kalmijn Nederlandse gemeenten: Een verklaring vanuit sociologisch en demografisch
perspectief. Maandstatistiek van de Bevolking, 47, 15-24. Nederlandse gemeenten: Een verklaring vanuit sociologisch en demografisch
perspectief. Maandstatistiek van de Bevolking, 47, 15-24. Graaf, P.M. de & M. Kalmijn (2006). Change and stability in the social determi-
nants of divorce: A comparison of marriage cohorts in the Netherlands. Euro-
pean Sociological Review, 22, 561-572. 95 Hannan, M.T. & N. Brandon Tuma (1990). A reassessment of the effect of income
maintenance on marital dissolution in the Seattle-Denver experiment. Ameri-
can Journal of Sociology, 95, 1270-1298. Herrnstein, R.J. & C. Murray (1994). The Bell Curve: Intelligence and class structure in
American life. New York: Free Press. Hoem, J.M. (1997). Educational gradients in divorce risks in Sweden in recent
decades. Population Studies, 51, 19-28. Kalmijn, M., A. Loeve & D. Manting (2007). Income dynamics of couples and the
dissolution of marriage and cohabitation. Demography, 44, 159-179. Kalmijn, M. & A.-R. Poortman (2006). His or her divorce? A study of divorce deter-
minants using information on who initiated the divorce. European Sociological
Review, 22, 201-214. Kephart, W.M. & T.P. Monahan (1952). Desertion and divorce in Philadelphia. age selection: Prospective evidence for reciprocal effects of marital status and
health. Social Science and Medicine, 43, 113-123. Zuckerman, J.T. (1950). A socio-legal approach to family desertion. Marriage and
Family Living, 12, 83-88. matthijs kalmijn Literatuur Ame-
rican Sociological Review, 17, 719-727. Lamont, M. (1992). Money, morals, and manners: The culture of the French and the Ame-
rican upper-middle class. Chicago: The University of Chicago Press. Leeuwen, M.H.D. van, I. Maas & A. Miles (2002). HISCO. Historical International
Standard Classification of Occupations. Leuven: Leuven University Press. Maas, I. & M.H.D. van Leeuwen (2004). HISCO HISCLASS recode scheme. http://histo-
ryofwork.iisg.nl/. Poortman, A.-R. (2005). How work affects divorce: The mediating role of financial
and time pressures. Journal of Family Issues, 26, 168-195. Poppel, F. van (1992). Trouwen in Nederland: Een historisch-demografische stu-
die van de 19e en vroeg-20e eeuw. Wageningen: Landbouwuniversiteit Wage-
ningen. Poppel, F. van & J. de Beer (1993). Measuring the effect of changing legislation
on the frequency of divorce: The Netherlands, 1830-1990. Demography 30, 425-
441. Prescott, C.A. & K.S. Kendler (2001). Associations between marital status and alco-
hol consumption in a longitudinal study of female twins. Journal of Studies on
Alcohol, 62, 589-604. Ross, C.E. & C. Wu. (1995). The links between education and health. American Soci-
ological Review, 60, 719-745. South, S.J. (2001). Time-dependent effects of wives’ employment on marital dis-
solution. American Sociological Review, 66, 226-245. Teachman, J.D. (2002). Stability across cohorts in divorce risk factors. Demography,
39, 331-352. Waldron, I., M.E. Hughes & T.L. Brooks (1996). Marriage protection and marri- voorlopers in de echtscheidingsrevolutie 96 96 matthijs kalmijn matthijs kalmijn Vroege ontkerkelijking in Nederland
Een analyse van het geboortecohort 1850-1882 Hans Knippenberg en Sjoerd de Vos Hans Knippenberg en Sjoerd de Vos Inleiding en probleemstelling Nog voor het eind van de negen-
tiende eeuw werd de honderdduizend overschreden en in 1930 bleken al 1,1 mil-
joen Nederlanders niet tot enige kerk of godsdienstige gemeenschap gerekend te
willen worden. Het ging toen al om ruim veertien procent van de bevolking. Na
1930 vertraagde het proces enigszins zonder overigens tot stilstand te komen. De
volkstelling van 1947 – die van 1940 werd vanwege de oorlog en bezetting uitge-
steld – kwam uit op 1,6 miljoen onkerkelijken, overeenkomend met zeventien
procent van de bevolking. Tot 1960 vond een verdere vertraging plaats, maar na
de culturele revolutie van de jaren zestig is er geen houden meer aan en secu-
lariseert de Nederlandse samenleving in een adembenemend tempo. Volgens
de meest recente surveys – volkstellingen zijn er na 1971 niet meer gehouden –
behoort op dit moment 61 tot 66 procent van de bevolking niet tot enige kerk of
godsdienstige gemeenschap (Becker & De Hart, 2006; Bernts e.a., 2007; Sengers,
2005). 98 98 )
Weten we dankzij dergelijke surveys veel (maar lang niet alles) over de recen-
te ontkerkelijking, over de vroege (eind negentiende-, begin twintigste-eeuwse)
ontkerkelijking zijn we veel minder goed ingelicht, al beschikken we natuurlijk
wel over de inmiddels klassiek geworden studies van Kruijt (1933) en Staverman
(1954), die zich vooral op volkstellingen (van 1920 respectievelijk 1947) baseer-
den. Zowel surveys als volkstellingen hebben in het algemeen het nadeel dat
het momentopnames zijn en dat veranderingen gedurende de levensloop van
individuen grotendeels buiten beeld blijven. In dat opzicht biedt de Historische
Steekproef Nederlandse bevolking (HSN) unieke mogelijkheden, omdat dit data-
bestand in principe wel tracht individuen gedurende hun gehele levensloop te
volgen, inclusief het kerkgenootschap waartoe zij behoren. De tussenvoeging ‘in
principe’ moet hier wel serieus genomen worden, want de informatie over de
veranderingen in kerkgenootschap en de momenten waarop deze plaatsvonden
is, zoals we later nog zullen toelichten, niet volledig, en daarnaast kon lang niet
iedereen in deze steekproef van de wieg tot het graf gevolgd worden. Toch biedt de HSN uniek materiaal waar het gaat om vragen als hoe vaak
veranderde men van kerkgenootschap, welke overgangen vonden plaats, bij
welke gelegenheden vonden overgangen plaats, was bij godsdienstig gemengde
huwelijken het kerkgenootschap van de man of dat van de vrouw maatgevend,
enzovoorts. Binnen het kader van deze bijdrage beperken wij ons vooral tot de
overgang naar (en eventueel van) de onkerkelijkheid gedurende de periode 1850-
1940. Inleiding en probleemstelling Godsdienst heeft altijd een opvallende plaats in de Nederlandse samenleving
ingenomen. Traditioneel werden Nederlanders aangeduid als een volk van koop-
lieden en dominees. Busken Huet karakteriseerde de zeventiende-eeuwse inwo-
ners van de Republiek al als een ‘volk van theologanten’ (geciteerd in Kruijt,
1933, 1). Deze samenleving onderging vanaf het laatste kwart van de negentien-
de eeuw een structurele differentiatie in aparte rooms-katholieke en (orthodox-)
protestantse segmenten, die samen met socialistische en liberale ‘volksdelen’
later als de bekende vier zuilen van de Nederlandse samenleving aangeduid zou-
den worden. Ook elders in Europa kwam zuilvorming op basis van religie voor
(Hellemans, 1990, 1993; Lijphart, 1968, 1977; Righart, 1986). Toch is Nederland in
zoverre uniek dat een uitgewerkte protestantse (calvinistische) zuil, die boven-
dien in het verzuilingsproces het voortouw nam, naast een uitgewerkte katholie-
ke zuil, nergens anders voorkwam. Elders ging het meestal om een tegenstelling
tussen klerikalen (rooms-katholieken) en anti-klerikalen (liberalen, socialisten
of communisten). In Nederland heeft dat de ‘etnisering’ van katholieken en
orthodox-protestanten en daarmee het godsdienstig bewustzijn versterkt (Knip-
penberg, 1996; Knippenberg & Van der Wusten, 2001; Van Rooden, 1996). Tot de
jaren zestig van de twintigste eeuw zou deze verzuiling standhouden. Het ineen-
storten daarvan nadien zou gepaard gaan met een massale secularisatie in de
vorm van een sterk dalend kerklidmaatschap en kerkbezoek (Becker & De Hart,
2006; Becker & De Wit, 2000; Bernts, Dekker & De Hart, 2007; Knippenberg, 1992,
1998, 2005). Nederland geldt nu als een van de meest geseculariseerde landen
van de westerse wereld. Nederland is ‘domineesland voorbij’, om de titel van een
bundel beschouwingen over dit onderwerp te parafraseren (Berkhof e.a., 1988). De constante in deze toch wel spectaculaire ontwikkeling is dat Nederlan-
ders ook in hun onkerkelijkheid iets van hun traditionele godsdienstig bewust-
zijn overeind houden. Zodra men de kerk verlaten heeft, komt men daar rond voor uit. Ook in zijn of haar onkerkelijkheid heeft de Nederlander iets calvinis-
tisch. De omvang van deze bewuste onkerkelijkheid kon daarom al vroeg via
volkstellingen geregistreerd worden en bleek in het laatste kwart van de negen-
tiende eeuw substantiële vormen aan te nemen. Gaven bij de op last van koning
Lodewijk Napoleon in 1809 gehouden volkstelling nog slechts 295 mensen te
kennen niet tot een kerk te behoren, bij de volkstelling van 1879 waren dat er
al 12.253 op een bevolking van ruim vier miljoen (De Kok, 1964; Knippenberg,
1992). Daarna nam de onkerkelijkheid snel toe. Inleiding en probleemstelling Daarbij proberen we de volgende onderzoeksvragen te beantwoorden. hans knippenberg en sjoerd de vos 1. Wat was voor verschillende groepen personen (te onderscheiden op
basis van individuele en contextuele kenmerken) de kans onkerkelijk
te worden, dan wel (weer) kerkelijk te worden? Hoe veranderden deze
kansen in de loop van de onderzochte periode en gedurende de levens-
loop? 99 2. Wat was de invloed van cruciale gebeurtenissen zoals huwelijk en ver-
huizing op het verlaten van de kerk? 2. Wat was de invloed van cruciale gebeurtenissen zoals huwelijk en ver-
huizing op het verlaten van de kerk? Alvorens een poging te wagen deze vragen via het HSN-bestand te beantwoorden
ontwikkelen we in volgende paragraaf eerst een aantal hypothesen op grond van
de bestaande literatuur en bespreken we aansluitend de aard van de gebruikte
HSN-data en de methodische problemen die daarmee gepaard gaan. Theoretische gezichtspunten Algemene theorieën Theorieën over secularisering werden lange tijd gedomineerd door de verbin-
ding met moderniseringsprocessen, of het nu ging om van Max Weber afgeleide
inzichten over een toenemende rationaliteit (die Entzauberung der Welt), dan wel
om aan Durkheim ontleende ideeën over functionele differentiatie (zie voor een
recent overzicht van dit klassieke seculariseringsparadigma Bruce, 2002). Kruijts
studie (1933) over de vroege onkerkelijkheid in Nederland bevat in feite dezelfde
verklaringselementen, wanneer hij een relatie legt met ‘de rationalisering van
het economische leven’ (industrialisatie, ontwikkeling moderne technieken), de
ontwikkeling van de (natuur)wetenschap, verstedelijking, en het functieverlies
der kerken. Speciale aandacht bij hem krijgt de invloed van het socialisme als
een uitvloeisel van de met modernisering gepaard gaande sociale spanningen
(ibidem). Die sociale spanningen kunnen ook verklaren waarom in Nederland de
vroege onkerkelijkheid enerzijds vooral op het Friese en Groningse platteland,
en anderzijds in de vroeg geïndustrialiseerde Zaanstreek gevonden wordt (Knip-
penberg, 1992). Staverman (1954), die een speciale studie wijdde aan de vroege
Friese onkerkelijkheid, wijst op de grote maatschappelijke tegenstellingen en
grote sociale nood onder de landbouwende bevolking als gevolg van de grote
agrarische crisis (1878-1895), die velen de kerk de rug deden toekeren en hun
heil deden zoeken in het mede daardoor opkomende socialisme. Voor veel ‘soci-
aal ontevredenen’ vormde dit socialisme als het ware een alternatieve religie. Niet voor niets zou de eerste socialistische afgevaardigde in de Tweede Kamer,
Ferdinand Domela Nieuwenhuis, door het echtpaar Jan en Annie Romein (1973;
oorspronkelijk 1938-1940) later als ‘de apostel der arbeiders’ worden gekarakte-
riseerd en riepen zijn aanhangers ‘Us ferlosser komt!’ als hij op tournee was in
Friesland (Frieswijk, Kalma & Kuiper, 1988, 7). Zelf had Domela Nieuwenhuis, vroege ontkerkelijking in nederland oorspronkelijk opgeleid tot en werkzaam als luthers predikant, in 1879 de kerk
verlaten, omdat ‘niet het christenzijn, maar het menschzijn de hoofdzaak [was]’,
terwijl voor een kerkgenootschap het omgekeerde gold (geciteerd in Mönnich,
1988, 64-65). Bij de volkstelling van 1889 bleek de bevolking van het Friese kies-
district Schoterland, van waaruit Domela Nieuwenhuis in 1888 gekozen was, al
voor 16,3% onkerkelijk te zijn (Frieswijk, 1988). De gemeente Schoterland was
met 20,6% onkerkelijken zelfs de meest onkerkelijke gemeente van heel Neder-
land (Knippenberg, 1992). In het algemeen was de eerste socialistische beweging
fel anti-kerkelijk (Kruijt, 1928; Staverman, 1954). 100 j
(
j
)
Het klassieke seculariseringsparadigma veronderstelt ook een seculariseren-
de werking van de Reformatie op de langere termijn. 100 Theoretische gezichtspunten Die loopt via individuali-
sering (het protestantse principe van het algemeen priesterschap der gelovigen)
en daardoor ook fragmentering (er ontbreekt – in tegenstelling tot de rooms-
katholieke kerk – een autoriteit tussen God en individu) en alfabetisering (een
protestant moet zelf de Bijbel kunnen lezen). Ook de ‘protestantse ethiek’ die
industrieel kapitalisme en economische groei zou stimuleren, draagt daaraan
bij, alsmede de met de Reformatie gepaard gaande rationalisering in de vorm
van het ontmythologiseren van de wereld en het elimineren van rituele en sacra-
mentele manipulatie van God (Bruce, 2002). Dat verklaart ook waarom in het
algemeen protestanten eerder en meer seculariseren dan rooms-katholieken, en
waarom meer vrijzinnig-protestanten (met een grotere individuele vrijheid aan-
gaande de geloofsinhoud en kerkelijke normen) eerder en meer seculariseren
dan hun orthodoxere geloofsgenoten. Een tweede groep theorieën ontkent in feite dat er sprake zou zijn van een
voortgaand proces van secularisering gekoppeld aan de modernisering van de
samenleving (Stark, 1999). Vooral de ontwikkelingen in de Verenigde Staten,
dat als modern westers land een hoge mate van godsdienstigheid handhaafde,
bracht de bedenkers ervan op een marktbenadering van (on)kerkelijkheid (Finke,
Guest & Stark, 1996; Finke & Stark, 1988; Stark & Innacone, 1994). Waar moderni-
seringstheoretici de vraag naar kerk en godsdienst zagen afnemen, bijvoorbeeld
doordat seculiere instellingen behorende bij de verzorgingsstaat functies als
armenzorg, onderwijs, huisvesting, et cetera overnamen van kerkelijke instel-
lingen, en de ontwikkeling van wetenschap en kennis de rationaliteit van de
samenleving deden groeien, daar gingen deze godsdienstige markttheoretici
uit van een stabiele vraag naar godsdienstige ‘producten’. Verschillen in kerke-
lijkheid en onkerkelijkheid verklaarden zij dan ook via de aanbodzijde van de
godsdienstmarkt. Een sterke marktregulering (bijvoorbeeld door een staatskerk,
die de keuzevrijheid van individuen belemmert) en een geringe diversiteit van
het aanbod (waardoor competitie tussen de aanbieders ontbreekt) zou leiden tot
een dalende kerkelijkheid. Op basis van deze inzichten ontwikkelden zij zelfs
de hypothese dat stedelijke gebieden door hun diverser en toegankelijker aan-
bod godsdienstiger en kerkelijker zouden zijn dan plattelandsgebieden (Finke &
Stark, 1988). hans knippenberg en sjoerd de vos Recentelijk hebben Norris en Inglehart (2004) een nieuwe theorie over
secularisering ontwikkeld, die zij de theorie van existentiële zekerheid noe-
men. In hun wereldwijde analyse van de verschillen in godsdienstigheid tus-
sen landen vinden zij geen empirische steun voor een aanbodgerichte theorie
over secularisering (zie ook Wunder, 2005). Theoretische gezichtspunten Zij verwerpen dan ook het idee dat
er overal een stabiele vraag naar godsdienst zou zijn. Integendeel, in de meer
welvarende en ontwikkelde delen van de wereld vermindert de behoefte aan
godsdienst, wat in feite een ondersteuning van de klassieke seculariserings-
these inhoudt, maar elders blijft die behoefte op een constant hoog niveau of
stijgt die zelfs. Ter verklaring ontwikkelden zij de hypothese dat de behoefte
aan godsdienst van mensen afhankelijk is van de mate van bestaanszekerheid
gedurende hun ‘formative years’, dat wil zeggen de periode waarin ze opgroei-
en en gevormd worden tot volwassenen. Opgroeien in een omgeving met een
groot gebrek aan bestaanszekerheid zou dan leiden tot een relatief belang-
rijke plaats voor godsdienst in hun leven. Zo zou de sterke secularisering in
de (West-)Europese verzorgingsstaten verklaard kunnen worden door de hoge
mate van bestaanszekerheid aldaar. De godsdienstigheid van veel landen in het
Zuiden, maar zelfs ook van de VS met zijn veel minder ontwikkelde collectieve
verzorgingsarrangementen zou omgekeerd verklaard worden door een grotere
mate van existentiële onzekerheid. 101 De invloed van individuele kenmerken De invloed van individuele kenmerken Los van deze algemene theorieën over secularisering blijken bepaalde individu-
ele kenmerken van invloed te zijn op het al dan niet verlaten van de kerk. Zo blij-
ken mannen in het algemeen onkerkelijker te zijn dan vrouwen (Verweij, 1998). Opvallend genoeg geldt dat zowel voor de Nederlandse samenleving rond 1900
als rond 2000, al lijkt het verschil in het laatste kwart van de twintigste eeuw
beduidend kleiner te zijn geworden (Becker & De Hart, 2006; Faber & Ten Have,
1970; Knippenberg, 2001; Knippenberg & De Vos, 1991; Schepens, 1991). Ter ver-
klaring van dit verschil wordt wel een tweetal sociaal-economische theorieën
genoemd: de ‘workforce theory’ en de ‘child-rearing theory’ (Verweij, 1998). De
‘workforce theory’ (Hoge & Roozen, 1979; De Vaus, 1984) legt de relatie met een
verschillende participatie op de arbeidsmarkt. Hoe meer arbeid (buitenshuis)
verricht wordt, des te minder is er tijd en aanleiding om kerkelijk betrokken te
raken. En aangezien mannen meer buitenshuis werken en werkten dan vrouwen
zou hiermee het verschil verklaard zijn. De toegenomen arbeidsparticipatie van
vrouwen in het laatste kwart van de twintigste eeuw zou dan tevens het klei-
ner worden van het verschil kunnen verklaren. De ‘child-rearing theory’ (Hoge
& Roozen, 1979; De Vaus, 1982; De Vaus & McAllister, 1987) legt de nadruk op de
relatief grote betrokkenheid van vrouwen bij de zorg en opvoeding van kinde-
ren. Doordat kerken zich traditioneel (denk ook aan het Nederlandse verzuilde
onderwijs) ook op het vlak van onderwijs en opvoeding bewegen, raken moe- vroege ontkerkelijking in nederland ders eerder kerkelijk betrokken dan vaders. Andere theorieën wijzen op verschil-
lende persoonlijkheidskenmerken: vrouwen zouden angstiger en minder tot het
nemen van risico’s geneigd zijn, en in het algemeen socialer, gevoeliger, meer
betrokken bij het welzijn van anderen (Need & De Graaf, 2005). Anderen noemen
een grotere bereidheid van vrouwen zich over te geven aan een hogere macht,
een sterker schuldbesef, waarvoor vergeving gezocht moet worden, een sterkere
trouw aan eenmaal aangegane bindingen en een grotere behoefte aan leider-
schap (Verweij, 1998). 102 Leeftijd is een tweede individueel kenmerk dat in de literatuur genoemd
wordt als van invloed te zijn op onkerkelijkheid: jongeren zijn in het algemeen
onkerkelijker dan ouderen. Ook deze samenhang geldt zowel in de huidige
samenleving als in die van een eeuw geleden (Becker & De Hart, 2006; Faber &
Ten Have, 1970; Knippenberg, 2001; Knippenberg & De Vos, 1991; Verweij, 1998). De invloed van individuele kenmerken Samenhangen met leeftijd zijn altijd lastig te interpreteren (Cobben & Hagenaars,
1977; Hagenaars, 1977). Zij kunnen het gevolg zijn van een leeftijdseffect: naar-
mate men ouder wordt, gaat men zich anders gedragen. Onduidelijk is dan nog
welke met leeftijd samenhangende biologische, psychologische, sociologische of
economische verschijnselen de werkelijke achtergrond vormen. Samenhangen
met leeftijd kunnen echter ook een gevolg zijn van een zogenaamd cohorteffect:
eerder geboren cohorten gedragen zich (op alle leeftijden) anders dan later gebo-
ren cohorten, waardoor op een bepaald moment het beeld ontstaat dat oudere
mensen zich anders gedragen dan jongere. Voor zover het om ontkerkelijking
gaat is zowel sprake van een leeftijdseffect als van een cohorteffect, waarbij dat
laatste effect sterker is (Knippenberg & De Vos, 1991). Het leeftijdseffect houdt
in dat meer mensen in de loop van hun leven, dus naarmate ze ouder worden,
van kerkelijk onkerkelijk worden dan omgekeerd. Het cohorteffect houdt in dat
eerder geboren cohorten kerkelijker zijn en blijven dan later geboren cohorten. De verklaring daarvan moet vooral worden gezocht in verschillende waardeori-
ëntaties: eerder geboren cohorten houden er traditionelere waarden op na dan
later geboren cohorten. Omdat het cohorteffect sterker is dan het leeftijdseffect,
zijn op een bepaald moment jongeren in grotere mate onkerkelijk dan ouderen. Een derde individueel kenmerk dat vaak in verband gebracht wordt met
ontkerkelijking is opleidingsniveau (zie bijvoorbeeld Becker & De Hart, 2006; De
Graaf, Need & Ultee, 2000; Knippenberg & De Vos, 1991; Schepens, 1991). Hoog-
opgeleiden zijn in het algemeen onkerkelijker dan laagopgeleiden. Verklaringen
van dit verband zijn gebaseerd op een kritischer en onafhankelijker (minder
gezagsgetrouwe) geesteshouding en een grotere mate van (technische) kennis bij
hoger opgeleiden. Voor een groot deel overlappen dergelijke verklaringen met
het klassieke seculariseringsparadigma. Een vierde individueel kenmerk, dat ten dele samenhangt met het vorige, is
het beroep. Op basis van de volkstelling van 1930 constateerde Kruijt (1935) al dat
er grote verschillen in onkerkelijkheid tussen de beroepsgroepen/bedrijfsklassen
waren, die uiteenliepen van ruim 4 procent in de landbouw tot 27 procent in het hans knippenberg en sjoerd de vos krediet- en bankwezen. Ook Staverman (1954) en Kuiper (1953) kwamen later tot
vergelijkbare conclusies in hun analyses van de volkstelling van 1947. De invloed van individuele kenmerken Voor zover
zij zich aan theoretische beschouwingen op dit punt waagden, lagen deze min
of meer in lijn met het klassieke seculariseringsparadigma, waarbij industriali-
sering (relatief hoge onkerkelijkheid onder industriearbeiders), modernisering
(relatief hoge onkerkelijkheid onder de nieuwe middenstand) als ook opleidings-
niveau (relatief hoge onkerkelijkheid onder intellectuele en vrije beroepen) een
belangrijke rol spelen. 103 Hypothesen Lang niet alle bovenvermelde theoretische gezichtspunten zijn onmiddellijk te
vertalen naar het niveau van individuen. Vele hebben betrekking op gemeen-
schappen, van lokaal tot nationaal. De analyses worden daarom op twee manie-
ren uitgevoerd. Allereerst zullen we op basis van de HSN een aantal relevante
kenmerken van individuen selecteren, die direct of indirect afgeleid kunnen
worden uit bovengenoemde theoretische overwegingen. Daarbij zullen overigens
niet alle hiervoor genoemde aspecten aan de orde kunnen komen; zo ontbreken
in de HSN bijvoorbeeld gegevens met betrekking tot de opleiding. In de tweede
plaats kunnen we, door personen te koppelen aan hun woonplaats, proberen
kenmerken van een hoger schaalniveau, bijvoorbeeld gemeenten of provincies,
in de analyse te betrekken. Daartoe zullen HSN-gegevens via de woonplaats van
betrokkenen gekoppeld worden aan gegevens uit volkstellingen en eventueel
andere databestanden. We formuleren nu de volgende hypothesen op het individuele niveau: Leeftijd
In de loop van hun leven worden meer mensen onkerkelijk dan kerkelijk
(leeftijdseffect). Hoe later men geboren is, hoe groter de kans dat men onkerkelijk wordt
(cohorteffect). Geslacht
Mannen hebben een grotere kans om onkerkelijk te worden dan vrouwen. Aard van het oorspronkelijke kerkgenootschap
Geboren protestanten hebben een grotere kans om onkerkelijk te worden dan
geboren rooms-katholieken. Geboren leden van vrijzinnige kerkgenootschappen hebben een grotere kans
om onkerkelijk te worden dan geboren leden van meer rechtzinnige kerken. Opleiding/beroep
Personen met beroepen waarvoor een hoog opleidingsniveau is vereist Leeftijd
In de loop van hun leven worden meer mensen onkerkelijk dan kerkelijk
(leeftijdseffect). Hoe later men geboren is, hoe groter de kans dat men onkerkelijk wordt
(cohorteffect). Geslacht Geslacht Mannen hebben een grotere kans om onkerkelijk te worden dan vrouwen. Geboren protestanten hebben een grotere kans om onkerkelijk te worden dan
geboren rooms-katholieken. p
g/
p
Personen met beroepen waarvoor een hoog opleidingsniveau is vereist p
g/
p
Personen met beroepen waarvoor een hoog opleidingsniveau is vereist vroege ontkerkelijking in nederland vroege ontkerkelijking in nederland hebben een grotere kans om onkerkelijk te worden dan personen met
beroepen die weinig of geen opleiding vergen. hebben een grotere kans om onkerkelijk te worden dan personen met
beroepen die weinig of geen opleiding vergen. De rol van de omgeving is vertaald in de volgende hypothesen: 104 Verstedelijking
Personen in stedelijke omgevingen hebben een grotere kans om onkerke-
lijk te worden dan personen in rurale omgevingen (seculariseringspara-
digma). Personen in rurale omgevingen hebben een grotere kans om onkerkelijk te
worden dan personen in stedelijke omgevingen (religieuze marktheorie). Industrialisatie
Personen in industriële omgevingen hebben een grotere kans om onker-
kelijk te worden dan personen in niet-industriële, met name agrarische,
omgevingen (seculariseringsparadigma). Godsdienstige diversiteit
Personen in godsdienstig homogene omgevingen hebben een grotere kans
om onkerkelijk te worden dan personen in godsdienstig heterogene omge-
vingen (religieuze marktheorie). De registratie van godsdienst in de HSN De registratie van godsdienst in de HSN
Onze analyse steunt vooral op de registratie van godsdienst zoals die opgenomen
is in de gemeentelijke bevolkingsregisters. Bij bijzondere gebeurtenissen zoals
geboorte, huwelijk en verhuizing werd het kerkgenootschap geregistreerd op
basis van een opgave van de betrokkene zelf (huwelijk en verhuizing) of diens
ouders (geboorte). Ook kon iemand op eigen initiatief zijn kerkgenootschap
laten veranderen. Een tweede bron van mogelijke registratie bieden de tienjaar-
lijkse volkstellingen, die als belangrijke functie hadden de gemeentelijke bevol-
kingsregisters te controleren en zo nodig te corrigeren. Aangezien bij die volks-
tellingen gevraagd werd naar het kerkgenootschap waartoe men zichzelf achtte
te behoren (inclusief de optie ‘geen’), was het in principe mogelijk om de tien
jaar het kerkgenootschap van de betrokkene vast te stellen en zo nodig te ver-
anderen in het bevolkingsregister. Helaas zijn er aanwijzingen dat dit niet altijd
ook daadwerkelijk gebeurd is. Zo vermeldt Van Doorn (1990) dat in Den Haag
een eventuele verandering in kerkgenootschap alleen bij verhuizen werd gere-
gistreerd. Correctie via de volkstellingen zou daar dan op dit punt niet plaats
hebben gevonden. 105 Om na te gaan of de godsdienstgegevens vanuit de HSN redelijk betrouwbaar
zijn hebben we ze vergeleken met volkstellingsgegevens. Daartoe is een verge-
lijking gemaakt van de godsdienstige gezindte van overeenkomstige leeftijdsco-
horten in de HSN-steekproef in 1922 en de volkstelling van 1920 (zie tabel 1). Daarbij is bij de HSN bewust gekozen voor een later jaar, omdat aangenomen
mag worden dat er enige tijd verstreek voordat de bevolkingsregisters aangepast
werden aan de volkstellingsdata. We beperken ons tot de grootste kerken (her-
vormden en katholieken) en uiteraard de onkerkelijken. Tabel 1 laat zien dat er sprake is van een onderregistratie van onkerkelijken
in de HSN. Dit verschijnsel doet zich sterker voor in Rotterdam en de provincie
Utrecht dan in de provincies Friesland en Zeeland. Weliswaar is de vergelijking
niet helemaal correct, omdat in ons HSN-bestand in bijvoorbeeld de provincie
Utrecht alleen gegevens bekend zijn van mensen die geboren zijn in de provin-
cies Friesland, Zeeland en Utrecht, en in Rotterdam, terwijl de gegevens van de Tabel 1 De godsdienstige gezindten in de HSN in 1922 vergeleken met die van de overeen-
komstige leeftijdscohorten in de volkstelling van 1920 HSN 1922 Geboortecohort 1851-1880
VT 1920 Geboortecohort 1851-1880
% NH
% RK
% Onk. n
% NH
% RK
% Onk. De gebruikte steekproef Voor onze analyses is gebruikgemaakt van Historische Steekproef Nederlandse bevol-
king (HSN), dataset Levenslopen, release 2007.01. Hieruit selecteerden we alle perso-
nen die geboren zijn in de periode 1850-1882. Helaas was vanuit de HSN nog niet
van geheel Nederland voldoende informatie beschikbaar, met als gevolg dat de
analyses alleen betrekking hebben op personen die geboren zijn in de provincies
Friesland, Utrecht en Zeeland plus de stad Rotterdam inclusief latere annexa-
ties. Jammer genoeg blijft daardoor het homogeen katholieke zuiden buiten
beeld en ontbreekt bijvoorbeeld de mogelijkheid een vergelijking te maken tus-
sen katholieken onder en boven de Moerdijk. Onze conclusies hebben dan ook
slechts betrekking op een deel van Nederland. De steekproef bestaat uit 5.150
aselect getrokken onderzoekspersonen (zie ook de bijdrage van Mandemakers in
dit boek). Van 276 daarvan is in de HSN geen enkel gegeven over de godsdienstige
gezindte bekend, zodat onze analyses uiteindelijk betrekking hebben op 4.874
personen. Voor de toetsing van onze hypotheses over de rol van de omgeving maken
we gebruik van gegevens afkomstig uit de Historisch-Ecologische Databank (HED) van
de afdeling Geografie en Planologie van de Universiteit van Amsterdam en oor-
spronkelijk afkomstig uit de volkstelling en bedrijfstelling van 1930 (CBS Elfde
Algemene Volkstelling van 31 December 1930; CBS Bedrijfstelling 31 December
1930). hans knippenberg en sjoerd de vos De registratie van godsdienst in de HSN n
Friesland
57,8
4,9
8,2
367
56,4
6,7
9,5
100.660
Utrecht
47,8
37,7
1,7
297
48,1
30,5
5,1
83.018
Zeeland
56,7
22,3
1,6
314
54,8
25,1
2,4
62.728
Rotterdam
56,6
27,2
2,0
302
49,2
24,5
8,1
118.527
Bronnen: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82; HED. vroege ontkerkelijking in nederland volkstelling ook gebaseerd zijn op mensen die uit andere delen van Nederland
naar de provincie Utrecht zijn verhuisd, maar de verschillen tussen de HSN en
de volkstelling zijn zo groot dat ze daardoor niet volledig verklaard kunnen wor-
den. Als oorzaak van de verschillen kan genoemd worden dat gemeenten niet
altijd gebruikmaakten van de volkstellingen om de kerkelijke gezindte van hun
inwoners te corrigeren. Met name grote stedelijke gemeenten lijken op dit punt
in gebreke gebleven te zijn. Dat blijkt ook als binnen Friesland, waar de onder-
schatting van het aandeel onkerkelijken het minst is, apart gekeken wordt naar
Leeuwarden; daar is het percentage onkerkelijken volgens de HSN gelijk aan 4,3
en volgens de volkstelling aan 14,4. Bij de interpretatie van onze uitkomsten
dient met de, regioafhankelijke, onderschatting van de onkerkelijkheid reke-
ning gehouden te worden. 106 Aangezien de opgaven van de godsdienst van betrokkenen sinds de latere
jaren dertig van de twintigste eeuw ten gevolge van de overgang op persoons-
kaarten nog minder betrouwbaar zijn, hebben we geen meetpunten na 1940 in
onze analyses opgenomen; zelfs de gegevens van 1940 moeten met de nodige
voorzichtigheid geïnterpreteerd worden. Analyses van individuele kenmerken Jannie en Sietze Voor we proberen de ontkerkelijking van verschillende groepen personen nader
te analyseren buigen we ons eerst over de lotgevallen van twee individuen. Laten
we ze Jannie en Sietze noemen, maar dat zullen vast hun echte namen niet zijn. Jannie wordt in 1850 geboren in Rotterdam, als dochter in een Nederlands
hervormd gezin. In 1865 blijkt ze te behoren tot de Christelijk Afgescheidenen,
maar in 1872 is ze Apostolisch. In 1879 is ze teruggekeerd naar de Christelijk
Afgescheidenen, maar later in datzelfde jaar is ze zelfs weer Nederlands her-
vormd geworden. In 1900 is duidelijk dat ze is overgegaan naar de Gereformeer-
de Kerken, die ze tot haar dood in 1912 trouw blijft. Ook Sietze wordt geboren in een Nederlands hervormd gezin en wel in 1860
in Het Bildt in Friesland. Wanneer hij volwassen wordt verlaat hij deze kerk en
staat hij als onkerkelijk te boek. Op zijn 27ste geeft hij evenwel te kennen weer
Nederlands hervormd te zijn. Enkele jaren later wordt Sietze wederom onkerke-
lijk. Vervolgens treedt hij toe tot de Gereformeerde Kerken, die hij alweer spoedig
in 1899 verlaat om voor de derde maal onkerkelijk te worden. In 1920 blijkt hij
weer Nederlands hervormd te zijn, in 1921 weer Gereformeerd, en een jaar later
toch maar weer Nederlands hervormd. In 1926 wordt hij voor de vierde maal als
onkerkelijk geregistreerd. Negen jaar later blijkt hij weer Nederlands hervormd
te zijn, wat hij tot zijn dood in 1944 blijft. De lotgevallen van Jannie en Sietze zijn uitzonderlijk, maar zij illustreren
toch op zijn minst een paar zaken die tot nog toe in de literatuur onopgemerkt hans knippenberg en sjoerd de vos Tabel 2 Frequentie wijzigingen in kerkgenootschap (inclusief onkerkelijkheid) 10
Aantal wijzigingen
Aantal personen
Percentage
0
3.840
78,8
1
494
10,1
2
324
6,6
3
124
2,5
4
55
1,1
5
21
0,4
6
7
0,1
7
3
0,1
8
2
0,0
9
3
0,1
10
1
0,0
Totaal
4.874
100,0
Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. of in ieder geval onderbelicht zijn gebleven. Sommige mensen wisselden meer-
dere keren van kerk en gingen ook meerdere keren heen en weer tussen kerke-
lijk en onkerkelijk. Tabel 2 laat zien hoe dat bij alle onderzoekspersonen zat. Analyses van individuele kenmerken Daarbij is onder-
scheid gemaakt tussen 46 verschillende kerkgenootschappen, inclusief de optie
onkerkelijk; de tabel zou overigens niet veel anders zijn geweest als het aantal
kerkgenootschappen tot een stuk of tien zou zijn beperkt. Het blijkt dat 79 pro-
cent van hen tijdens hun levensloop niet van kerkgenootschap veranderde (voor
een deel omdat ze heel vroeg zijn overleden), dat tien procent één keer veran-
derde, bijvoorbeeld onkerkelijk werd, krap zeven procent twee keer veranderde,
krap drie procent drie keer, en de rest, krap twee procent, maar liefst vier tot tien
keer zijn of haar kerkgenootschap (inclusief de seculiere optie) wijzigde. Daarbij
moet men in gedachten houden dat het aantal geconstateerde wijzigingen een
onderschatting vormt van het werkelijke aantal, omdat, zoals we al eerder heb-
ben aangegeven, lang niet alle wijzigingen ook daadwerkelijk geregistreerd zul-
len zijn. Leeftijd en geslacht De onderzoekspersonen zijn onderverdeeld in drie geboortecohorten: zij die
geboren zijn in de periode 1853-1862 (verder aan te duiden als het cohort5362), zij
die geboren zijn in de periode 1863-1872 (cohort6372), en zij die geboren zijn in
de periode 1873-1882 (cohort7382). Van deze cohorten is op bepaalde tijdstippen
de mate van onkerkelijkheid vastgesteld. Deze tijdstippen zijn zodanig gekozen vroege ontkerkelijking in nederland Tabel 3 Percentage onkerkelijken van drie geboortecohorten gedurende de levensloop (tot
1940), totaal en vrouwen en mannen apart
1862
1872
1882
1892
1902
1912
1922
1932
1940
Totaal
Cohort5362 (1.004,107)a
0,0
0,2
0,7
1,8
1,6
2,0
1,8
4,4
5,6
Cohort6372 (1.225,396)
0,5
0,9
1,4
2,0
3,0
4,4
8,2
9,8
Cohort7382 (944,489)
1,0
1,5
2,3
4,5
6,1
7,4
9,0
Vrouwen
Cohort5362 (510,58)
0,0
0,2
0,5
2,9
1,9
2,2
1,3
3,8
6,9
Cohort6372 (595,210)
0,7
1,1
1,0
1,6
2,1
3,9
7,3
8,6
Cohort7382 (487,270)
0,8
1,1
2,4
2,9
5,4
5,2
7,8
Mannen
Cohort5362 (494,49)
0,0
0,2
0,8
0,6
1,3
1,9
2,3
5,2
4,1
Cohort6372 (630,186)
0,3
0,7
1,8
2,5
4,0
4,9
9,2
11,3
Cohort7382 (457,219)
1,1
2,0
2,2
6,2
7,0
10,1
10,5
a Bij de aanduidingen van de cohorten staan tussen haakjes de aantallen personen uit het eerste en het
laatste meetjaar vermeld. Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. 108 a Bij de aanduidingen van de cohorten staan tussen haakjes de aantallen personen uit het eerste en het
laatste meetjaar vermeld. Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. j
Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. dat eventuele veranderingen die in de bevolkingsregisters worden aangebracht
op basis van de voorafgaande volkstelling de tijd hebben gehad om doorgevoerd
te worden. Het gaat steeds om het einde van het betreffende jaar. In tabel 3 staat
achter de aanduiding van het cohort vermeld hoeveel mensen er in het eerste
meetjaar (1862, 1872 of 1882) deel van uit maakten, en hoeveel daarvan in 1940
nog in leven zijn; deze strategie wordt ook in veel andere tabellen toegepast. dat eventuele veranderingen die in de bevolkingsregisters worden aangebracht
op basis van de voorafgaande volkstelling de tijd hebben gehad om doorgevoerd
te worden. Het gaat steeds om het einde van het betreffende jaar. Leeftijd en geslacht In tabel 3 staat
achter de aanduiding van het cohort vermeld hoeveel mensen er in het eerste
meetjaar (1862, 1872 of 1882) deel van uit maakten, en hoeveel daarvan in 1940
nog in leven zijn; deze strategie wordt ook in veel andere tabellen toegepast. Tabel 3 maakt duidelijk dat beide hypothesen over de samenhang met leef-
tijd worden bevestigd. Het leeftijdseffect is aanwezig, omdat voor alle drie de
cohorten geldt dat ze onkerkelijker worden naarmate de levensloop vordert. Zo
blijken degenen die geboren werden tussen 1853 en 1862 in dat laatste jaar alle-
maal kerkelijk te zijn, maar blijken degenen van dat cohort die in 1940 nog in
leven zijn al voor 5,6 procent onkerkelijk te zijn. Het cohort 1863-1872 is dan al
voor bijna 10 procent onkerkelijk, ondanks het feit dat ze in 1940 10 jaar jonger
zijn. Ook het derde cohort is in 1940 aanzienlijk onkerkelijker dan bij de geboor-
te: 9 tegen 1 procent. Bij de eerste twee cohorten vindt vooral in de periode 1922-
1932 een sterke toename in onkerkelijkheid plaats. Bovendien is er sprake van een cohorteffect: de onderzoekspersonen van een
bepaald cohort blijken steeds onkerkelijker dan het voorgaande cohort tien jaar
eerder (dat wil zeggen op dezelfde leeftijd). Meestal zijn ze zelfs onkerkelijker
dan het voorgaande cohort op hetzelfde moment, wat erop wijst dat het cohort-
effect sterker is dan het leefijdseffect. hans knippenberg en sjoerd de vos Dezelfde analyse is ook voor mannen en vrouwen apart uitgevoerd (zie tabel
3). Zowel bij de mannen als bij de vrouwen is sprake van een leeftijdseffect en
een cohorteffect. De relatief sterke toename in onkerkelijkheid in de periode
1922-1932 blijkt zich bij de mannen in alle drie cohorten voor te doen, terwijl dat
bij de vrouwen alleen voor de eerste twee cohorten geldt. Tenslotte wordt ook de
hypothese over de rol van geslacht bevestigd: we zien het bekende verschijnsel
dat mannen in het algemeen onkerkelijker zijn dan vrouwen, al blijkt dat voor
het oudste cohort niet helemaal op te gaan. 109 Oorspronkelijk kerkgenootschap
Welke kerkgenootschappen seculariseerden het meest? Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. a Bij de aanduidingen van de godsdienstige gezindte staan tussen haakjes de aantallen personen uit het
eerste en het laatste meetjaar vermeld. Leeftijd en geslacht Om deze vraag te beant-
woorden hebben we verschillende kleinere kerkgenootschappen moeten samen-
Tabel 4 Percentage onkerkelijken gedurende de levensloop naar oorspronkelijk kerkgenoot-
schap en geboortecohort
1862
1872
1882
1892
1902
1912
1922
1932
1940
Cohort 1853-1862
Hervormd
(687,80)a
0,0
0,2
0,8
2,4
2,1
2,4
2,2
5,9
6,3
Katholiek
(222,17)
0,0
0,5
0,0
0,0
0,0
0,0
0,0
0,0
0,0
Gereformeerd
(26,5)
0,0
0,0
0,0
0,0
0,0
0,0
0,0
0,0
0,0
Luth/Rem/Doop
(49,3)
0,0
0,0
0,0
0,0
0,0
5,6
0,0
0,0
0,0
Israëlitisch
(17,1)
0,0
0,0
0,0
0,0
0,0
0,0
0,0
0,0
0,0
Cohort 1863-1872
Hervormd
(842,284)
0,4
0,8
1,8
2,6
3,5
5,3
10,8
12,3
Katholiek
(275,81)
0,0
0,0
0,0
0,5
1,6
1,8
1,5
2,5
Gereformeerd
(51,15)
0,0
0,0
0,0
0,0
3,1
0,0
0,0
0,0
Luth/Rem/Doop
(42,15)
2,4
7,9
5,6
3,0
3,3
8,0
10,0
13,3
Israëlitisch
(9,0)
0,0
0,0
0,0
0,0
0,0
0,0
0,0
-
Cohort 1873-1882
Hervormd
(666,343)
0,6
1,7
2,9
6,2
7,2
8,2
10,2
Katholiek
(189,106)
0,0
0,0
0,0
0,0
2,2
2,4
3,8
Gereformeerd
(40,20)
0,0
0,0
0,0
0,0
8,7
0,0
0,0
Luth/Rem/Doop
(26,11)
3,8
4,8
5,3
5,3
5,9
21,4
18,2
Israëlitisch
(17,4)
0,0
0,0
0,0
0,0
7,1
15,4
25,0
a Bij de aanduidingen van de godsdienstige gezindte staan tussen haakjes de aantallen personen uit het
eerste en het laatste meetjaar vermeld. Oorspronkelijk kerkgenootschap Oorspronkelijk kerkgenootschap
Welke kerkgenootschappen seculariseerden het meest? Om deze vraag te beant-
woorden hebben we verschillende kleinere kerkgenootschappen moeten samen- bel 4 Percentage onkerkelijken gedurende de levensloop naar oorspronkelijk kerkgenoot-
schap en geboortecohort Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. vroege ontkerkelijking in nederland voegen, omdat anders de aantallen te klein werden om er zinvolle uitspraken
over te doen; zelfs nu zijn de aantallen soms nog erg klein. De verschillende gere-
formeerde kerkgenootschappen zijn om die reden samengevoegd evenals de wat
vrijzinniger protestantse kerken van lutheranen, remonstranten en doopsgezin-
den. De (paar) Waals hervormden zijn bij de Nederlands hervormden gevoegd, en
de (paar) oud-katholieken bij de rooms-katholieken. De twee Israëlitische kerk-
genootschappen (Nederlands en Portugees) zijn samengevoegd; het gaat daarbij
overigens nog steeds om zeer kleine aantallen. Per cohort worden de verschil-
lende kerkgenootschappen vergeleken. 110 De secularisering van het oudste cohort blijkt zich vrijwel uitsluitend in her-
vormde kringen te hebben afgespeeld (zie tabel 4). Slechts één katholiek en één
doopsgezinde hebben de kerk (tijdelijk) de rug toegekeerd in de periode tot 1940. Leeftijd en geslacht In die periode neemt onder de oorspronkelijk hervormden het percentage onker-
kelijken toe tot ruim zes procent. De secularisering van het middelste cohort is
duidelijk al verder voortgeschreden en beperkt zich niet langer tot vrijwel alleen
hervormden. Ook de gecombineerde groep van lutheranen, remonstranten en
doopsgezinden seculariseert, al gaat het steeds maar om enkele mensen. Voor
het eerst verlaten ook katholieken hun kerk, beginnend rond de eeuwwisseling. De gereformeerden en ‘Israëlieten’ blijven hun kerk het meest trouw. Het jong-
ste cohort levert in het algemeen een vergelijkbaar beeld op als het voorgaande
cohort. Maar voor het eerst zien we ook secularisering onder de joodse kerkge-
nootschappen, al gaat het daarbij om slechts twee mensen. De resultaten voor alle drie de cohorten zijn in overeenstemming met de
eerder gestelde hypothesen dat protestanten meer seculariseren dan katholie-
ken, en dat de ontkerkelijking sterker is bij de meer vrijzinnige dan bij de meer
rechtzinnige kerken. Interessant is ook de vraag naar het omgekeerde traject: hoeveel personen,
die als onkerkelijk begonnen zijn, hebben uiteindelijk toch kerkelijk onderdak
gevonden? Daarbij gaat het om slechts zeer weinig mensen; in de drie cohor- el 5 Routes naar onkerkelijkheid en eventueel weer terug van geborenen tussen 1850 en
1882 per oorspronkelijk kerkgenootschap (in procenten) Tabel 5 Routes naar onkerkelijkheid en eventueel weer terug van geborenen tussen 1850 en
1882 per oorspronkelijk kerkgenootschap (in procenten) Aantal
Ooit onkerkelijk
Onkerkelijk
geëindigd
Terug naar
totaal
rechtstreeks
via andere
oorsprong
andere
Hervormd
3.339
7,0
6,1
0,9
4,2
1,9
0,9
Katholiek
1.092
1,3
1,1
0,2
1,1
0,2
0,0
Gereformeerd
173
2,9
1,7
1,2
0,0
0,6
2,3
Luth/Rem/Doop
183
7,1
4,9
2,2
3,3
0,5
3,3
Israëlitisch
70
4,3
4,3
0,0
4,3
0,0
0,0
Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. hans knippenberg en sjoerd de vos hans knippenberg en sjoerd de vos ten zijn achtereenvolgens slechts twee, drie en vijf personen onkerkelijk gestart. Niettemin valt op dat zeven van hen later toch kerkelijk zijn geworden, waarvan
vijf tot en met 1940 dan wel het moment van overlijden toe. Dat ondersteunt
het al eerdergenoemde verschijnsel dat kerkverlating zeker niet altijd definitief
was. 111 Om meer inzicht te krijgen in de routes vanuit de verschillende kerkge-
nootschappen naar de onkerkelijkheid en eventueel terug is tabel 5 gevormd
op basis van alle onderzoekspersonen geboren tussen 1850 en 1882. Leeftijd en geslacht Deze tabel
laat opnieuw zien dat de vroege onkerkelijkheid vooral veroorzaakt werd door
het verlaten van een protestantse kerk. Binnen het protestantse volksdeel waren
het vooral de leden van de calvinistische moederkerk en haar oude protestantse
zusters (lutherse en doopsgezinde kerken), die relatief veel onkerkelijk werden. Dat gebeurde overwegend rechtstreeks, maar de route kon ook via andere ker-
ken lopen. Opvallend is dat veel van de onkerkelijk geworden protestanten tij-
dens hun leven toch weer kerkelijk werden door terug te keren naar hun oor-
spronkelijke kerk of onderdak te vinden bij een ander kerkgenootschap. Binnen
het protestantse volksdeel waren de verschillende soorten gereformeerden het
minst geneigd tot ontkerkelijking. Het meest opvallende is wellicht dat van
deze ooit onkerkelijk geworden gereformeerden niemand onkerkelijk eindigde. De meesten vonden uiteindelijk onderdak bij een andere kerk, een minderheid
keerde terug naar de oorspronkelijke kerk. Van alle kerkgenootschappen waren
de katholieken het minst geneigd hun kerk te verlaten. Slechts 1,3 procent liet
zich ooit als onkerkelijk registreren. In tegenstelling tot de gereformeerden en
in mindere mate ook de leden van de andere protestantse kerkgenootschappen
hadden zij evenwel een geringe neiging weer kerkelijk te worden als zij eenmaal
onkerkelijk geworden waren. Om meer inzicht te krijgen in de routes vanuit de verschillende kerkge-
nootschappen naar de onkerkelijkheid en eventueel terug is tabel 5 gevormd
op basis van alle onderzoekspersonen geboren tussen 1850 en 1882. Deze tabel
laat opnieuw zien dat de vroege onkerkelijkheid vooral veroorzaakt werd door
het verlaten van een protestantse kerk. Binnen het protestantse volksdeel waren
het vooral de leden van de calvinistische moederkerk en haar oude protestantse
zusters (lutherse en doopsgezinde kerken), die relatief veel onkerkelijk werden. Beroep en opleidingsniveau Tabel 6 toont de relatie tussen beroepsstatus en (on)kerkelijkheid. Voor de
beroepsstatus is gebruikgemaakt van de HISCAM schaal (zie daarvoor Maas, Lam-
bert, Zijdeman, Prandy & Van Leeuwen, 2006 en Prandy, 2000). De hoogst moge-
lijke status bij deze schaal is 100 en de laagste in theorie 0, maar in de praktijk
10. Aangezien er in 1862 en 1872 geen onkerkelijken met beroep in de steekproef
zaten en in 1882 slechts drie, zijn deze jaren buiten beschouwing gelaten. Omdat
van de mannen een groter deel een beroep had dan van de vrouwen, spelen zij
in de tabel een grotere rol. Het aantal onkerkelijken is in het begin erg klein,
daarom staat in deze tabel voor alle duidelijkheid bij alle gemiddeldes voor deze
groep het aantal mensen aangegeven waarop het gemiddelde is gebaseerd. In de tijd blijkt zich een opmerkelijke omslag voor te doen. In de jaren 1892
en 1902 is de gemiddelde beroepsstatus van de onkerkelijken lager dan van de ker-
kelijken, terwijl vanaf 1912 het omgekeerde geldt. De verschillen in gemiddelde
beroepsstatus tussen kerkelijken en onkerkelijken in de jaren 1892 en 1902 zijn vroege ontkerkelijking in nederland Tabel 6 Gemiddelde beroepsstatus van kerkelijken en onkerkelijken, geboortecohort 1850-
1882, gedurende de levensloop 1892
1902
1912
1922
1932
1940
Kerkelijk
(933,403)a 41,5
45,2
47,8
49,8
49,6
49,9
Onkerkelijk
41,0 (16)
43,6 (29)
51,1 (46)
51,6 (50)
55,5 (53)
54,6 (41)
a Bij de aanduiding kerkelijk staan tussen haakjes de aantallen personen uit het eerste en het laatste
meetjaar vermeld; bij de onkerkelijken worden ze bij elk meetjaar vermeld. Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. 112 a Bij de aanduiding kerkelijk staan tussen haakjes de aantallen personen uit het eerste en het laatste
meetjaar vermeld; bij de onkerkelijken worden ze bij elk meetjaar vermeld. Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. gering en niet significant, maar passen wel mooi bij het klassenkarakter van
de vroege onkerkelijkheid dat we eerder bespraken en dat in analyses van de
geografische spreiding van de onkerkelijkheid in het laatste kwart van de negen-
tiende eeuw en begin van de twintigste eeuw naar voren kwam (Knippenberg,
1992; Kruijt, 1933; Staverman, 1954). Het waren de gebieden met sterke sociale
tegenstellingen, zoals de zuidoosthoek van Friesland en de Zaanstreek, waar
socialisme en onkerkelijkheid in onderlinge verbondenheid voet aan de grond
kregen. Beroep en opleidingsniveau Het is aardig dat verschijnsel hier op individueel niveau terug te zien,
al kennen we niet de politieke voorkeur van betrokkenen. Datzelfde geldt voor
de relatieve toename van de onkerkelijkheid onder werkenden met een hogere
status in de eerste helft van de twintigste eeuw, wanneer die onkerkelijkheid
een grotere omvang begint te krijgen en ook geografisch herkenbaar wordt door
de toename in de luxere suburbane gemeenten als Blaricum, Hilversum, Bloe-
mendaal, Bussum, Heemstede en Wassenaar (Knippenberg, 1992). Deze vorm van
onkerkelijkheid is te beschouwen als de voorloper van de massale onkerkelijk-
heid die zich vanaf de jaren zestig van de vorige eeuw gaat manifesteren, waar-
bij welstand en een hoog opleidingsniveau een individualistische mentaliteit en
levensstijl bevorderen, die vaak in strijd komt met traditionele geloofswaarden
en kerkelijke verplichtingen. Overigens geldt dat de gemiddelde beroepsstatus
van kerkelijken in de jaren vanaf 1912 weliswaar duidelijk hoger ligt dan die van
de onkerkelijken, maar het verschil is alleen in 1932 significant bij een onbe-
trouwbaarheid van vijf procent, en dan zelfs bij één procent. Enige voorzichtig-
heid met de conclusies is dus op zijn plaats. gering en niet significant, maar passen wel mooi bij het klassenkarakter van
de vroege onkerkelijkheid dat we eerder bespraken en dat in analyses van de
geografische spreiding van de onkerkelijkheid in het laatste kwart van de negen-
tiende eeuw en begin van de twintigste eeuw naar voren kwam (Knippenberg,
1992; Kruijt, 1933; Staverman, 1954). Het waren de gebieden met sterke sociale
tegenstellingen, zoals de zuidoosthoek van Friesland en de Zaanstreek, waar
socialisme en onkerkelijkheid in onderlinge verbondenheid voet aan de grond
kregen. Het is aardig dat verschijnsel hier op individueel niveau terug te zien,
al kennen we niet de politieke voorkeur van betrokkenen. Datzelfde geldt voor
de relatieve toename van de onkerkelijkheid onder werkenden met een hogere
status in de eerste helft van de twintigste eeuw, wanneer die onkerkelijkheid
een grotere omvang begint te krijgen en ook geografisch herkenbaar wordt door
de toename in de luxere suburbane gemeenten als Blaricum, Hilversum, Bloe-
mendaal, Bussum, Heemstede en Wassenaar (Knippenberg, 1992). Deze vorm van
onkerkelijkheid is te beschouwen als de voorloper van de massale onkerkelijk-
heid die zich vanaf de jaren zestig van de vorige eeuw gaat manifesteren, waar-
bij welstand en een hoog opleidingsniveau een individualistische mentaliteit en
levensstijl bevorderen, die vaak in strijd komt met traditionele geloofswaarden
en kerkelijke verplichtingen. Beroep en opleidingsniveau Overigens geldt dat de gemiddelde beroepsstatus
van kerkelijken in de jaren vanaf 1912 weliswaar duidelijk hoger ligt dan die van
de onkerkelijken, maar het verschil is alleen in 1932 significant bij een onbe-
trouwbaarheid van vijf procent, en dan zelfs bij één procent. Enige voorzichtig-
heid met de conclusies is dus op zijn plaats. Om dit verschijnsel in onze steekproef verder te analyseren zijn de beroepen
ingedeeld in het klassenschema HISCLASS (zie Van Leeuwen & Maas, 2005) en
vervolgens verder samengevoegd in vijf categorieën. Daarbij is zoveel mogelijk
acht geslagen op het opleidingsniveau, omdat aparte gegevens over het oplei-
dingsniveau, afgezien van de (on)geletterdheid van gehuwden, in de HSN ont-
breken. Tabel 7 geeft het resultaat. Daarin wordt onderscheid gemaakt tussen
hoofdarbeiders en handarbeiders, en daarbinnen tussen hoge, gemiddelde en
lage scholing en ongeschoold. Hooggeschoolde handarbeiders ontbreken. Het beeld is duidelijk, en in overeenstemming met de bevindingen op basis hans knippenberg en sjoerd de vos Tabel 7 Het percentage onkerkelijken per beroepsgroep, geboortecohort 1850-1882, gedu-
rende de levensloop Tabel 7 Het percentage onkerkelijken per beroepsgroep, geboortecohort 1850-1882, gedu-
rende de levensloop Tabel 7 Het percentage onkerkelijken per beroepsgroep, geboortecohort 1850-1882, gedu-
rende de levensloop 1892
1902
1912
1922
1932
1940
Hoofdarbeid/hooggeschoold
(19,15)a
0,0
0,0
10,8
8,6
22,2
26,7
Hoofdarbeid/rest
(142,89)
0,7
2,1
5,8
6,2
9,3
9,0
Handarbeid/middelgeschoold
(209,145)
2,9
1,4
2,9
4,0
7,3
11,0
Handarbeid/laaggeschoold
(305,83)
1,6
3,3
3,3
6,9
5,9
7,2
Handarbeid/ongeschoold
(274,112)
1,5
2,5
3,1
3,7
6,5
7,1
a Bij de aanduidingen van de beroepsgroep staan tussen haakjes de aantallen personen uit het eerste
en het laatste meetjaar vermeld. Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. 113 a Bij de aanduidingen van de beroepsgroep staan tussen haakjes de aantallen personen uit het eerste
en het laatste meetjaar vermeld. Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. van tabel 6. Aanvankelijk blijken de onkerkelijken verhoudingsgewijs meer bij
de handarbeiders dan bij de hoofdarbeiders aangetroffen te worden, maar vanaf
1912 is het omgekeerde het geval. Bij zowel de hoofd- als de handarbeiders halen
geleidelijk aan de hoger opgeleiden hogere onkerkelijkheidspercentages dan de
lager opgeleiden. van tabel 6. Aanvankelijk blijken de onkerkelijken verhoudingsgewijs meer bij
de handarbeiders dan bij de hoofdarbeiders aangetroffen te worden, maar vanaf
1912 is het omgekeerde het geval. Bij zowel de hoofd- als de handarbeiders halen
geleidelijk aan de hoger opgeleiden hogere onkerkelijkheidspercentages dan de
lager opgeleiden. Beroep en opleidingsniveau Al met al verdient de eerder gestelde hypothese over de rol van beroep en
opleiding een bijstelling. De hypothese klopt wel in de latere jaren, vanaf 1912,
maar niet in de eerdere jaren, waarin juist laagopgeleiden een relatief hoge mate
van onkerkelijkheid kennen. Samenvatting van de analyses op het individuele niveau Samenvatting van de analyses op het individuele niveau We vatten de analyses op het individuele niveau nu samen met behulp van een
logistische-regressieanalyse, waarbij het al of niet onkerkelijk zijn in 1932 wordt
verklaard op basis van het geslacht, de leeftijd, de beroepsstatus en het oorspron-
kelijke kerkgenootschap. Er is voor dit jaartal gekozen, omdat de onkerkelijkheid
dan enig niveau heeft bereikt en zich ook in verschillende kerkgenootschappen
voordoet. Van het geboortecohort 1850-1882 zijn dan nog 1545 personen in leven,
van wie er slechts 710 een beroep hebben. Daarom hebben we twee analyses uit-
gevoerd: één zonder de variabele beroepsstatus en één met deze variabele. Bij
het oorspronkelijk kerkgenootschap zijn in de analyses vier groepen onderschei-
den: hervormd, katholiek, luthers/remonstrants/doopsgezind en Israëlitisch. De
gereformeerden zijn uit de analyses gehouden, omdat van hen in 1932 niemand
onkerkelijk was, hetgeen bij hen moeilijk interpreteerbare coëfficiënten zou heb-
ben opgeleverd; de verklaringskracht van de modellen zou overigens bij het wel
opnemen van deze groep nog iets groter zijn geweest. De overige godsdienstige
groepen waren te klein om een zinvolle analyse mogelijk te maken. Tabel 8 geeft
allereerst de resultaten inclusief beroepsstatus. In deze analyse zijn 679 perso-
nen opgenomen, van wie er 53 onkerkelijk zijn. In de tabel staan bij geslacht vroege ontkerkelijking in nederland en oorspronkelijke godsdienst de aantallen personen met de betreffende eigen-
schap vermeld. p
Tabel 8 laat zien dat de oorspronkelijke godsdienst de belangrijkste verkla-
rende variabele is. Personen die oorspronkelijk protestants zijn (zowel de her-
vormden als de gecombineerde groep lutheranen/remonstranten/doopsgezinden)
hebben een aanzienlijk grotere kans onkerkelijk te worden dan katholieken. Dat
lijkt ook voor Israëlieten te gelden, maar dit verschil is niet significant. Overigens
zijn de katholieken op hun beurt onkerkelijker dan de niet in de analyse opgeno-
men gereformeerden, die in 1932 immers allen kerkelijk zijn. Als tweede factor
komt leeftijd naar voren: jongeren zijn onkerkelijker dan ouderen. Eerder zagen
we al dat deze samenhang de resultante is van twee tegengestelde effecten: een
leeftijdseffect (naarmate men ouder wordt neemt de kans op onkerkelijkheid toe)
en een cohorteffect (later geboren cohorten zijn onkerkelijker dan eerder geboren
cohorten). Het cohorteffect blijkt dus sterker te zijn dan het leeftijdseffect. Als
derde factor komt de beroepsstatus naar voren: hoe hoger de status, des te groter
de kans onkerkelijk te worden. Deze samenhang is echter net niet significant bij
een onbetrouwbaarheid van vijf procent. 114 Samenvatting van de analyses op het individuele niveau De samenhang met geslacht is duidelijk
niet significant, al is de richting van het verband wel in overeenstemming met
onze hypothese dat mannen een grotere kans hebben onkerkelijk te worden dan
vrouwen. Hierbij moet bedacht worden dat in deze analyse alleen mannen en
vrouwen met beroep zijn opgenomen. De totale verklaringskracht van het model,
gemeten via de pseudo-R2 van Nagelkerke, is gelijk aan 0,095. 114 Tabel 8 Logistische-regresssieanalyse ter verklaring van het al of niet onkerkelijk zijn in 1932
van het geboortecohort 1850-1882, apart voor personen met een beroep en voor het
totale cohort Tabel 8 Logistische-regresssieanalyse ter verklaring van het al of niet onkerkelijk zijn in 1932
van het geboortecohort 1850-1882, apart voor personen met een beroep en voor het
totale cohort Personen met beroep
Totaal
n
b-coëfficiënt
Significantie
n
b-coëfficiënt
Significantie
Geslacht
Vrouw (ref.)
113
789
Man
566
0,422
0,393
689
0,489
0,017
Leeftijd
-0,045
0,026
-0,022
0,084
Beroepsstatus
0,018
0,051
Oorspronkelijke godsdienst
Katholiek (ref.)
159
330
Hervormd
495
2,061
0,005
1082
1,822
0,000
Luthers/remonstrant/
doopsgezind
15
2,473
0,019
47
2,122
0,001
Israëlitisch
10
1,827
0,158
19
2,002
0,023
Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. hans knippenberg en sjoerd de vos Het model zonder beroepsstatus heeft een geringere verklaringskracht: de
pseudo-R2 is nu gelijk aan 0,061; overigens is deze waarde slecht vergelijkbaar
met die van het vorige model, omdat er nu veel meer personen een rol spelen. Ook in dit model komt het oorspronkelijk kerkgenootschap als duidelijk de
belangrijkste factor naar voren. Verder blijken mannen nu bij een onbetrouw-
baarheid van vijf procent wel significant onkerkelijker te zijn dan vrouwen. De
invloed van leeftijd is nu echter niet meer significant. 115 Omgevingskenmerken: verstedelijking, industrialisatie en
godsdienstige diversiteit Voor de toetsing van onze hypotheses over de rol van de omgeving maken we
gebruik van gegevens afkomstig uit de volkstelling en de bedrijfstelling van
1930. We voeren de analyses uit op het niveau van gemeenten. De afhankelijke
variabele is het percentage van de bevolking dat onkerkelijk is. Er zijn vier ver-
klarende variabelen. Voor de mate van verstedelijking zijn twee indicatoren gehanteerd. Allereerst
de bevolkingsomvang van de grootste bewoonde kern van de gemeente. Aan dit
criterium voor stedelijkheid is de voorkeur gegeven boven de bevolkingsomvang
van de gemeente als geheel. Een aantal gemeenten, vooral in Friesland, beschikt
immers over een grote, doch zeer verspreid wonende, bevolking die geenszins
als stedelijk kan worden aangemerkt. Het percentage van de bevolking dat werk-
zaam is in de agrarische sector vormt een tweede, functioneler criterium voor
stedelijkheid. Het percentage van de bevolking dat werkzaam is in de industrie is
begrijpelijkerwijze gehanteerd om de mate van industrialisatie aan te geven. Voor de vaststelling van de godsdienstige diversiteit van de gemeente als een
indicatie voor de diversiteit in het aanbod aan verschillende kerkgenootschap-
pen is de volgende procedure gebruikt. De volkstelling onderscheidt, naast de
onkerkelijken en enkele restcategorieën, twaalf godsdienstige gezindten. Voor
de levensvatbaarheid van een godsdienstige gezindte in een gemeente stellen
we een grens van minimaal 25 aanhangers. De door ons gehanteerde maat voor
de diversiteit is gelijk aan het aantal van die twaalf godsdienstige gezindten met
minstens 25 aanhangers. In tabel 9 staan de onderlinge correlaties tussen de vier verklarende varia-
belen en het percentage onkerkelijken. De correlaties zijn gebaseerd op 1.076
gemeenten. Ze zijn allemaal significant bij een onbetrouwbaarheid van 1%. Verstedelijking blijkt positief samen te hangen met de mate van onkerke-
lijkheid. De correlatie met de bevolkingsomvang van de grootste kern in de
gemeente is positief en die met de omvang van de agrarische sector negatief. Ook met de mate van industrialisatie blijkt onkerkelijkheid positief samen te
hangen. Deze samenhangen zijn in overeenstemming met het klassieke secula-
riseringsparadigma, waarbij secularisering vooral in relatie wordt gebracht met
moderniseringsprocessen. Aangezien de concurrerende markttheorieën eerder vroege ontkerkelijking in nederland Tabel 9 Correlaties tussen kenmerken van gemeenten op basis van de volkstelling en bedrijfs-
telling 1930 % Onkerkelijk
Bevolkingsomvang
% Agrarisch
% Industrie
Bevolkingsomvang
0,219
% Agrarisch
-0,398
-0,239
% Industrie
0,285
0,151
-0,872
Diversiteit
0,578
0,399
-0,591
0,479
Bron: HED. Omgevingskenmerken: verstedelijking, industrialisatie en
godsdienstige diversiteit 116 een negatieve samenhang met verstedelijking voorspellen, vinden deze theorie-
ën geen steun in onze data. Dat wordt nog duidelijker wanneer specifiek naar
de samenhang met kerkelijke diversiteit gekeken wordt. Waar markttheorieën
een negatieve samenhang voorspellen – een divers aanbod leidt immers niet
alleen tot een ‘voor elk wat wils’-situatie, maar ook tot grotere concurrentie tus-
sen de aanbieders – blijkt hier sprake te zijn van een duidelijk positieve correla-
tie. De aan de markttheorie ontleende hypothesen dienen dan ook verworpen te
worden. Tezamen verklaren genoemde vier omgevingsvariabelen, die overigens
onderling soms zeer sterk samenhangen, 34,8 procent van de variantie van de
mate van onkerkelijkheid. een negatieve samenhang met verstedelijking voorspellen, vinden deze theorie-
ën geen steun in onze data. Dat wordt nog duidelijker wanneer specifiek naar
de samenhang met kerkelijke diversiteit gekeken wordt. Waar markttheorieën
een negatieve samenhang voorspellen – een divers aanbod leidt immers niet
alleen tot een ‘voor elk wat wils’-situatie, maar ook tot grotere concurrentie tus-
sen de aanbieders – blijkt hier sprake te zijn van een duidelijk positieve correla-
tie. De aan de markttheorie ontleende hypothesen dienen dan ook verworpen te
worden. Tezamen verklaren genoemde vier omgevingsvariabelen, die overigens
onderling soms zeer sterk samenhangen, 34,8 procent van de variantie van de
mate van onkerkelijkheid. Ondanks de correlaties met de omgevingskenmerken is er nog geen zeker-
heid dat er ook werkelijk sprake is van invloed van de omgeving. Zijn de mensen
in stedelijke gemeenten nu onkerkelijker, omdat ze door de stedelijke omgeving
in die richting worden gedreven, of zijn ze onkerkelijker, omdat ze andere indi-
viduele eigenschappen hebben? Hier zou juist een combinatie van gegevens van
de volkstelling en van de HSN een zeer nuttige rol kunnen spelen. We zouden
dan bijvoorbeeld kunnen nagaan of onder onderzoekspersonen van een bepaald
beroepsniveau die in een stad wonen relatief meer onkerkelijken voorkomen
dan onder onderzoekspersonen van hetzelfde beroepsniveau die op het platte-
land wonen. Ook zou het mogelijk zijn na te gaan of bijvoorbeeld protestanten
meer of minder gevoelig zijn voor de invloed van de (stedelijke) omgeving dan
katholieken (zie bijvoorbeeld Knippenberg & De Vos, 1989; De Vos, 1997). Helaas
konden deze analyses niet zinvol worden uitgevoerd. Dat is een gevolg van het al
eerder vermelde feit dat in de HSN sprake is van onderschatting van de onkerke-
lijkheid, en wel vooral in de stedelijke gebieden. hans knippenberg en sjoerd de vos hans knippenberg en sjoerd de vos Effect van cruciale gebeurtenissen tijdens de levensloop: huwelijk
en verandering van woonplaats Huwelijk 117 Een mogelijke reden om van kerk te veranderen dan wel deze te verlaten vormt
een huwelijk met iemand van een andere kerk of zonder kerkelijke binding. Bij alle onderzoekspersonen uit het geboortecohort 1850-1882 komen in totaal
1.577 eerste huwelijken voor, waarvan de kerkelijke gezindte van beide partners
bekend is. Tabel 10 geeft een overzicht uitgesplitst naar kerkelijke gezindte. Bij
de bepaling van het al of niet gemengd zijn van een huwelijk is weer gebruikge-
maakt van de indeling in 46 kerkgenootschappen waarop ook tabel 2 was geba-
seerd. In tabel 10 staan alleen de grotere godsdienstige groeperingen apart ver-
meld; de overige godsdiensten zijn samengevoegd tot ‘overigen’, maar wel met
de kanttekening dat hun huwelijken alleen ongemengd zijn als beide partners
precies dezelfde overige godsdienst hadden. Van alle 1.577 eerste huwelijken samen blijkt bijna 84 procent godsdienstig
ongemengd te zijn. Zo zijn er 908 huwelijken tussen partners die beide Neder-
lands hervormd zijn; het betreft dus 1.816 personen. Daarnaast zijn er 239 perso-
nen, onderzoekspersonen of juist de partner daarvan, die Nederlands hervormd
zijn maar getrouwd zijn met iemand van een andere godsdienstige gezindte
(zie voor een nadere uitsplitsing daarvan tabel 11). Voor de interpretatie van de
resultaten is het van belang zich bewust te zijn van het feit dat naarmate de Tabel 10 Ongemengde en gemengde huwelijken naar kerkelijke gezindte, geboortecohort
1850-1882 (alleen eerste huwelijken)
Aantal onge
mengde
huwelijken
Personen in
ongemengde
huwelijken
Personen in
gemengde
huwelijken
% Personen in
ongemengde
huwelijken
Nederlands Hervormd
908
1.816
239
88,4
Rooms-Katholiek
280
560
98
85,1
Gereformeerde Kerken
64
128
63
67,0
Christelijk Afgescheiden
19
38
10
79,2
Nederlands Israëlitisch
14
28
3
90,3
Doopsgezind
11
22
33
40,0
Geen
6
12
23
34,3
Overigen
17
34
47
42,0
Totaal
1.319
2.638
516
83,6
% van het totaal aantal huwelijken
83,6
Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-1882. bel 10 Ongemengde en gemengde huwelijken naar kerkelijke gezindte, geboortecohort
1850-1882 (alleen eerste huwelijken) vroege ontkerkelijking in nederland godsdienstige groep groter is ook de kans om met een lid van de eigen groep te
trouwen toeneemt. Dus dat hervormden een grote kans hebben met hervorm-
den te trouwen ligt alleen daarom al voor de hand. Des te opvallender zijn de
relatief grote kansen van rooms-katholieken, gereformeerden, afgescheidenen
en vooral joden om met een lid van de eigen godsdienstige gezindte te trouwen. Effect van cruciale gebeurtenissen tijdens de levensloop: huwelijk
en verandering van woonplaats Dat wijst bij deze groepen op sterke sociale grenzen met de buitenwereld. De in
deze periode van de grond komende verzuiling van de Nederlandse samenleving
is daar ongetwijfeld mede debet aan. Mede door hun kleine omvang trouwt de
groep onkerkelijken in meerderheid met een kerkelijke partner. Tabel 11 laat
zien welke varianten gemengde huwelijken in welke frequentie voorkomen. 118 In ruim eenderde van de gemengde huwelijken gaat het om een huwelijk
tussen een hervormde en een katholiek. Dat is niet verrassend als de omvang
van beide groepen verdisconteerd wordt, maar wel opmerkelijk gezien de relatief
grote godsdienstige ‘afstand’ tussen beide denominaties. De andere gemengde Tabel 11 De frequentie van de verschillende gemengde huwelijken, geboortecohort 1850-1882
(alleen eerste huwelijken)
Aantal
Percentage
Nederlands hervormd & rooms-katholiek
91
35,3
Nederlands hervormd & gereformeerde kerken
59
22,9
Nederlands hervormd & christelijk afgescheiden
8
3,1
Nederlands hervormd & Nederlands Israëlitisch
2
0,8
Nederlands hervormd & doopsgezind
27
10,5
Nederlands hervormd & geen
16
6,2
Nederlands hervormd & overige
36
14,0
Rooms-katholiek & doopsgezind
2
0,8
Rooms-katholiek & geen
1
0,4
Rooms-katholiek & overige
4
1,6
Gereformeerde kerken & geen
4
1,6
Christelijk afgescheiden & doopsgezind
1
0,4
Christelijk afgescheiden & overige
1
0,4
Nederlands Israëlitisch & overige
1
0,4
Doopsgezind & geen
1
0,4
Doopsgezind & overige
2
0,8
Geen & overige
1
0,4
Overige onderling
1
0,4
Totale aantal gemengde huwelijken
258
100,0
Percentage van het totaal aantal huwelijken
16,4
Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. el 11 De frequentie van de verschillende gemengde huwelijken, geboortecohort 1850-1882
(alleen eerste huwelijken) hans knippenberg en sjoerd de vos huwelijken betreffen vooral huwelijken tussen hervormden en leden van andere
protestantse kerken, zoals verschillende soorten gereformeerden en doopsge-
zinden. In de gemengde huwelijken met onkerkelijken blijkt de godsdienstige
partner vooral van protestantse huize te zijn. Slechts één huwelijk tussen een
katholiek en een onkerkelijke kwam in deze steekproef voor. 119 De gemengde huwelijken met een onkerkelijke partner zijn nog nader geana-
lyseerd om te zien wat er verder mee gebeurt en of hun eventuele kinderen als
kerkelijk of onkerkelijk worden geregistreerd. Vooraf zij benadrukt dat het om
kleine aantallen gaat, wat de generaliseerbaarheid van de uitkomsten minder
betrouwbaar maakt. Van de 22 gemengde huwelijken met een onkerkelijke part-
ner (één huwelijk viel af omdat het godsdienstig verloop ervan onduidelijk was)
blijft in tien gevallen hun godsdienstige gezindte ongewijzigd. Effect van cruciale gebeurtenissen tijdens de levensloop: huwelijk
en verandering van woonplaats In twaalf gevallen
neemt een van de twee de godsdienstige voorkeur van de ander over; acht maal
wint de kerkelijke variant en tien maal de gezindte van de vrouw. Deze huwelij-
ken worden dus alsnog godsdienstig homogeen en de kinderen krijgen (alsnog)
de gezindte van hun ouders. In de tien huwelijken die godsdienstig heterogeen
bleven, werden de kinderen in zes gevallen als onkerkelijk en in drie gevallen
als kerkelijk opgegeven; daarbij werd vijf keer de voorkeur van de vrouw en vier
keer die van de man gevolgd. In één geval van een gereformeerde man met een
onkerkelijke vrouw werd eerlijk gedeeld: hun eerste kind werd bij de geboorte
als onkerkelijk aangegeven, hun tweede kind als gereformeerd, hun derde weer
als onkerkelijk en hun vierde weer als gereformeerd. Deze ontwikkelingen en eventuele aanpassingen vonden plaats na de huwe-
lijksdatum. Dat doet de vraag rijzen of er ook al vóór die datum aanpassingen
hebben plaatsgevonden. Daarom is nagegaan wat de godsdienst van de gehuwde
onderzoekspersonen was voordat ze in het huwelijk traden; van de partners van
de onderzoekspersonen ontbreekt dat gegeven meestal. Bij alle zes homogeen
onkerkelijke huwelijken bleek dat de onderzoekspersoon bij de laatst beschik-
bare informatie vóór het huwelijk kerkelijk was (vijf maal Nederlands hervormd
en een maal gereformeerd) en zich dus mogelijk aangepast had aan zijn of haar
onkerkelijke partner, maar het kan natuurlijk ook zijn dat hij/zij al eerder onker-
kelijk was geworden zonder dat dit geregistreerd was. Verder komt het tien keer
voor dat bij een ongemengd kerkelijk huwelijk de onderzoekspersoon bij de laat-
ste meting vóór het huwelijk onkerkelijk was: zes keer bij een ongemengd Neder-
lands hervormd huwelijk, twee keer bij een ongemengd gereformeerd huwe-
lijk, een keer bij een ongemengd rooms-katholiek huwelijk en een keer bij een
ongemengd doopsgezind huwelijk. Zo lijkt ook aanpassing aan de godsdienstige
gezindte van de toekomstige partner vóór het huwelijk regelmatig voorgekomen
te zijn. vroege ontkerkelijking in nederland Verandering van woonplaats Verandering van woonplaats
Het achterliggende idee bij de invloed van verandering van woonplaats op ont-
kerkelijking is dat mensen die hun vertrouwde omgeving (inclusief de kerkelijke
gemeenschap met eventueel aanwezige sociale controle) verlaten eerder de kans
lopen daarmee ook afstand te doen van hun kerkelijke binding. Dit zal eerder
plaatsvinden naarmate de afstand tot de oorspronkelijke woonplaats groter is. a Bij de aanduidingen van de provincie staan tussen haakjes de aantallen personen uit het eerste en het
laatste meetjaar vermeld. Effect van cruciale gebeurtenissen tijdens de levensloop: huwelijk
en verandering van woonplaats We hebben er daarom voor gekozen het effect van het verlaten van de provin-
cie nader te analyseren en niet bijvoorbeeld het overschrijden van slechts de
gemeentegrens. Tabel 12 laat de mate van ontkerkelijking zien van het geboorte-
cohort 1850-1882 uitgesplitst naar het antwoord op de vraag of zij nog in hun
oorspronkelijke geboorteprovincie wonen, dan wel inmiddels naar een andere
provincie zijn verhuisd. 120 Er is sprake van een duidelijk patroon. In de beginperiode zijn mensen die
nog steeds in dezelfde provincie wonen onkerkelijker, maar daarna ontstaat het
verwachte beeld dat mensen die hun banden met de oude omgeving hebben ver-
loren door naar een andere provincie te verhuizen onkerkelijker zijn dan men-
sen die nog steeds in hun geboorteprovincie wonen. Weliswaar is het verschil
alleen significant in 1892 en in 1932 (bij een onbetrouwbaarheid van 5 procent),
maar daarmee is de omslag wel aangetoond, en de consistentie in de begin- en
de eindperiode is veelzeggend. Hierbij rijst wel de vraag of er wellicht sprake is
van selectieve migratie in die zin dat migranten andere kenmerken hebben dan
achterblijvers die mogelijkerwijze het verschil in de mate van onkerkelijkheid
kunnen verklaren. Om dat na te gaan hebben we de logistische-regressieanalyse
van tabel 8 uitgebreid met een verklarende variabele die aangeeft of de onder-
zoekspersoon in 1932 al of niet in een andere provincie woont dan bij zijn of
haar geboorte. Ook in dat model, waarin rekening is gehouden met de invloed
van geslacht, leeftijd, beroepsstatus en oorspronkelijk kerkgenootschap, blijken
de onderzoekspersonen die verhuisd zijn naar een andere provincie een grotere
kans te hebben om onkerkelijk te zijn; de bijbehorende b-coëfficiënt is gelijk aan
0,467. De samenhang is echter gezien de overschrijdingskans van 0,146 nu niet
significant. Maar ook als we deze, niet significante, samenhang serieus nemen Tabel 12 Percentage onkerkelijken uitgesplitst naar de vraag of men in dezelfde dan wel in
een andere provincie woont als de geboorteprovincie, geboortecohort 1850-1882 1872
1882
1892
1902
1912
1922
1932
1940
Zelfde
(2.223, 698)a
0,4
0,9
1,7
2,1
3,1
3,8
6,1
8,0
Andere
(114, 295)
0,0
0,0
0,2
1,7
3,6
5,8
10,0
11,2 a Bij de aanduidingen van de provincie staan tussen haakjes de aantallen personen uit het eerste en het
laatste meetjaar vermeld. Bron: HSN dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. Effect van cruciale gebeurtenissen tijdens de levensloop: huwelijk
en verandering van woonplaats hans knippenberg en sjoerd de vos hoeft de onkerkelijkheid niet het gevolg te zijn van de verhuizing; het kan ook
zijn dat de samenhang een gevolg is van het feit dat mensen die ertoe neigen
met de godsdienst te breken eerder naar een andere provincie verhuizen dan
mensen die daar niet toe neigen. Deze laatste mogelijke verklaring liet zich met
onze data uiteraard niet toetsen. 121 Conclusies Aanvankelijk was ontkerkelij- vroege ontkerkelijking in nederland king een puur hervormde aangelegenheid, later kwamen daar de meer vrijzin-
nige protestantse kerkgenootschappen als doopsgezinden, remonstranten en
lutheranen, en ook de joodse kerkgenootschappen bij. Uiteindelijk seculariseer-
den deze meer vrijzinnige protestanten minstens zo sterk als de hervormden, die
zoals bekend zowel meer vrijzinnige als meer orthodoxe modaliteiten verenig-
den (zie bijvoorbeeld Rasker, 1986). De verschillende soorten gereformeerden
ontkerkelijkten het minst. Het oorspronkelijk kerkgenootschap bleek de beste
voorspeller van ontkerkelijking op individueel niveau te zijn. king een puur hervormde aangelegenheid, later kwamen daar de meer vrijzin-
nige protestantse kerkgenootschappen als doopsgezinden, remonstranten en
lutheranen, en ook de joodse kerkgenootschappen bij. Uiteindelijk seculariseer-
den deze meer vrijzinnige protestanten minstens zo sterk als de hervormden, die
zoals bekend zowel meer vrijzinnige als meer orthodoxe modaliteiten verenig-
den (zie bijvoorbeeld Rasker, 1986). De verschillende soorten gereformeerden
ontkerkelijkten het minst. Het oorspronkelijk kerkgenootschap bleek de beste
voorspeller van ontkerkelijking op individueel niveau te zijn. 122 De samenhang tussen beroep en onkerkelijkheid liet een opmerkelijke
omslag zien. Tot en met 1902 ontkerkelijkten de categorieën met een lagere sta-
tus meer dan die met een hogere; datzelfde gold voor handarbeiders in vergelij-
king met hoofdarbeiders en voor lager of niet-geschoolden in vergelijking met
hoger geschoolden. Dit ondersteunt de visie van Kruijt (1933) dat in die begin-
periode ontkerkelijking en socialisme hand in hand gingen en juist vele ‘sociaal
ontevredenen’ hun kerk verlieten. Vanaf 1912 was het omgekeerde het geval en
ontkerkelijkten de categorieën met een hogere status, hoofdarbeiders en hoger
geschoolden meer dan respectievelijk de categorieën met een lagere status,
handarbeiders en lager of niet geschoolden. Deze soort van ontkerkelijking kan
gezien worden als een voorloper van de massale ontkerkelijking vanaf de jaren
zestig. Aangaande het effect van cruciale gebeurtenissen tijdens de levensloop op
mogelijke ontkerkelijking kan vastgesteld worden, dat godsdienstig gemengde
huwelijken met katholieken, gereformeerden en joden, hun omvang in aanmer-
king genomen, weinig voorkwamen, wat op relatief sterke sociale grenzen van
deze groepen met andere godsdienstige groeperingen wijst. Gemengde huwelij-
ken met een onkerkelijke partner kwamen bij katholieken nauwelijks voor en
waren overwegend protestant/onkerkelijk gemengd. Van deze laatste categorie
bleef na de huwelijksvoltrekking de helft trouw aan het eigen kerkgenootschap,
terwijl bij de andere helft aanpassing aan de partner plaatsvond, waarbij de ker-
kelijke gezindte van de vrouw meestal de doorslag gaf. Mede daardoor won de
kerkelijke partner in tweederde van de gevallen. Conclusies In het laatste kwart van de negentiende en het begin van de twintigste eeuw
werd godsdienst de belangrijkste scheidslijn in het maatschappelijk leven (De
Rooy, 1995). Onafhankelijk van elkaar trokken Van Rooden (1996) en Knippenberg
(1996) de parallel met processen van ‘etnisering’ waar katholieken en (orthodox-)
protestanten aan onderhevig zouden zijn. Dat betekende dat de sociale grenzen
tussen de verschillende godsdienstige groepen scherper afgebakend werden en
moeilijker te overschrijden. Maar dat betekende niet dat iedereen van de wieg
tot het graf zijn of haar kerk trouw bleef, zoals onze analyse van het geboorte-
cohort 1850-1882 duidelijk laat zien. Tijdens de levensloop kon men meermalen
van kerk veranderen, al vormden onze ‘Jannie en Sietze’ daarbij wel uitzonde-
ringen. Ruim 21 procent van onze steekproef uit genoemd geboortecohort veran-
derde minstens een keer in hun leven van kerkgenootschap of werd onkerkelijk;
11 procent overkwam dat zelfs minstens twee keer. Van belang daarbij was dat
het aantal kerkgenootschappen toenam. Dat gold vooral het protestantse volks-
deel. Nadat al eerder de Afscheiding van 1834 het godsdienstig pallet van nieuwe
(gereformeerde) kleuren had voorzien, voegde de Doleantie van 1886 daar nog
weer een nieuwe kleur aan toe. Bovendien bereikten nieuwe protestantse stro-
mingen ons land, waarvan baptisten, verschillende apostolische kerken, pink-
stergemeenten en het Leger des Heils getalsmatig de belangrijkste waren (Knip-
penberg, 1992). Op den duur getalsmatig de belangrijkste ‘gezindte’ zouden echter diegenen
vormen die zich niet (langer) tot een kerk of godsdienstige groepering wilden
rekenen. Werd bij het geboortecohort 1853-1863 nog iedereen met een kerkelij-
ke gezindte geboren, in het volgende cohort (1863-1873) was dat niet langer het
geval. Nadien zou het percentage onkerkelijk geborenen alleen maar toenemen. Bovendien werden een in de tijd toenemend aantal mensen tijdens hun levens-
loop alsnog onkerkelijk. Het zijn vooral de kenmerken van deze onkerkelijken,
die in onze analyse centraal stonden. Wij baseerden ons daarbij op een aselecte
steekproef uit personen die geboren waren in de provincies Friesland, Utrecht en
Zeeland en in de stad Rotterdam. Op grond daarvan konden wij concluderen dat
later geboren cohorten meer onkerkelijk werden dan eerder geboren cohorten,
dat de mate van onkerkelijkheid toenam naarmate de cohorten ouder werden,
en dat mannen een grotere kans hadden onkerkelijk te worden dan vrouwen. De
ontkerkelijking onder rooms-katholieken was in dit geboortecohort nog gering,
maar nam toe naarmate men later was geboren. Conclusies Kinderen werden dan kerkelijk
opgevoed. Bij huwelijken die godsdienstig gemengd bleven, lijken de kinderen
eerder als onkerkelijk dan als kerkelijk geregistreerd te worden, maar de aantal-
len zijn te klein om er betrouwbare uitspraken over te doen. Betrouwbaarder uitkomsten leverde onze analyse van het effect van verhui-
zing naar buiten de eigen provincie op. Tot en met 1902 zijn de achterblijvers
onkerkelijker, nadien zijn de migranten onkerkelijker. Aangezien in de eerste
periode de trek van het platteland naar de stad overheerst, is het helaas niet
uitgesloten dat de vermoedelijke onderregistratie van onkerkelijken in de grote
steden mede verantwoordelijk is voor de wat onverwachte uitkomsten in de peri-
ode tot en met 1902. Anderzijds past deze aanvankelijk relatief hoge onkerke-
lijkheid van de achterblijvers wel in het beeld van de vroege onkerkelijkheid op
het Friese platteland, waar Domela Nieuwenhuis en anderen hun socialistische
‘religie’ predikten. hans knippenberg en sjoerd de vos Naast individuele kenmerken en cruciale gebeurtenissen tijdens de levens-
loop bleken ook omgevingsfactoren van invloed op de kans onkerkelijk te wor-
den. Uit onze analyse op basis van gegevens uit de volkstelling van 1930 bleek
dat moderniseringsindicatoren zoals de mate van industrialisatie en verstede-
lijking (zowel positief via bevolkingsomvang van de grootste kern als negatief
via het percentage dat werkzaam was in de landbouw) in het algemeen positief
samenhingen met de mate van ontkerkelijking. De uit markttheorieën afgeleide
hypothese dat een divers godsdienstig aanbod ontkerkelijking zou kunnen voor-
kómen vond geen steun in deze gemeentelijke gegevens. Integendeel, godsdien-
stige diversiteit hing positief samen met de mate van onkerkelijkheid, wat zelfs
van de hier geselecteerde variabelen de sterkste samenhang met onkerkelijkheid
opleverde. 123 Dat brengt ons tenslotte weer terug bij de algemene seculariseringstheo-
rieën waarmee we deze bijdrage begonnen. Voor zover onze data daar uitsluit-
sel over kunnen geven, ondersteunen de uitkomsten toch vooral het klassieke
seculariseringsparadigma, dat ontkerkelijking in verband brengt met moderni-
seringsprocessen. Zowel de uitkomsten op individueel niveau (protestant versus
katholiek; vrijzinnig versus orthodox-protestant; (hoog)geschoold versus laag-/
niet-geschoold) als op gemeentelijk niveau (verstedelijking; industrialisatie;
godsdienstige diversiteit) ondersteunen dit paradigma. Daar komt bij dat de
enorme toename van de onkerkelijkheid in deze periode, die gepaard ging met
een toenemend aanbod aan met name protestantse kerkgenootschappen nu niet
bepaald wijst in de richting die de godsdienst-markttheoretici veronderstellen. Literatuur Becker, J. & J. de Hart (2006). Godsdienstige veranderingen in Nederland. Den Haag:
Sociaal en Cultureel Planbureau. Becker, J. & J. de Hart (2006). Godsdienstige veranderingen in Nederland. Den Haag:
Sociaal en Cultureel Planbureau. Becker J.W. & J.S.J. de Wit (2000). Secularisatie in de jaren negentig. Den Haag: Soci-
aal en Cultureel Planbureau. Berkhof, H. e.a. (1988). Voorbij domineesland. De gevolgen van de secularisatie. Amers-
foort/Leuven/Amsterdam: De Horstink/Trouw. Bernts, T., G. Dekker & J. de Hart (2007). God in Nederland 1996-2006. Kampen: Ten
Have. Bruce, S. (2002). God is dead. Secularization in the West. Oxford: Blackwell. Cobben, N.P. & J.A.P. Hagenaars (1977). Enkele analysetechnieken voor het bepa-
len van de effecten van leeftijd, kohort en periode. Sociale Wetenschappen, 20,
65-101. Doorn, M. van (1990). Haagse straatportretten. In Jaarboek 1990 Geschiedkundige
Vereniging Die Haghe (pp. 195-229). ’s-Gravenhage. Faber, H. & T.T. ten Have (1970). Ontkerkelijking en buitenkerkelijkheid in Nederland,
tot 1960. Assen: Van Gorcum. Finke, R., A. Guest & R. Stark (1996). Mobilizing local religious markets: religious
pluralism in the empire state. American Sociological Review, 63, 761-766. vroege ontkerkelijking in nederland Finke, R. & R. Stark (1988). Religious choice and competition. American Sociological
Review, 63, 761-766. Frieswijk, J. (1988). Welke schouders droegen Domela Nieuwenhuis naar de
Kamer? Een historisch-politicologische studie naar zijn verkiezing in 1888 in
Schoterland. In J. Frieswijk e.a. (red.), Ferdinand Domela Nieuwenhuis. De Apostel
van de Friese Arbeiders (pp. 131-149). Drachten/Leeuwarden: Friese Pers Boeke-
rij. 124 Frieswijk, J., J.J. Kalma. & Y. Kuiper (red.) (1988), Ferdinand Domela Nieuwenhuis. De
Apostel van de Friese arbeiders. Drachten/Leeuwarden: Friese Pers Boekerij. Graaf, N.D. de, A. Need & W. Ultee (2000). Levensloop en kerkverlating: een nieu-
we overkoepelende verklaring voor enkele empirische regelmatigheden. Mens
& Maatschappij, 75, 229-257. Hagenaars, J.A.P. (1977). Leeftijd, kohort en periode: een algemeen model voor de
analyse van sociale veranderingen. Sociale Wetenschappen, 20, 32-64. Hellemans, S. (1990). Strijd om de moderniteit. Sociale bewegingen en verzuiling in Euro-
pa sinds 1800. Leuven: Universitaire Pers. Hellemans, S. (1993). Zuilen en verzuiling in Europa. In U. Becker (red.), Neder-
landse politiek in historisch en vergelijkend perspectief (pp. 121-150). Amsterdam:
Het Spinhuis. Hoge, D.R. & D.A. Roozen (red.) (1979). Understanding church growth and decline,
1950-1978. New York: The Pilgrim Press. Knippenberg, H. (1992). De religieuze kaart van Nederland. Assen: Van Gorcum. Knippenberg, H. (1996). Nationale integratie en de ‘etnisering’ van katholieken
en protestanten: de rol van onderwijs. In H. Literatuur te Velde & H. Verhage (red.), De
eenheid en de delen. Zuilvorming, onderwijs en natievorming in Nederland 1850-1900
(pp. 177-196). Amsterdam: Het Spinhuis. Knippenberg, H. (1998). Secularization in the Netherlands in its historical and
geographical dimensions. GeoJournal, 45, 209-220. Knippenberg, H. (2001). Polarisatie en versnippering: kerk en godsdienst rond
1900. In J.G.S.J. van Maarseveen & P.K. Doorn (red.), Nederland een eeuw geleden
geteld (pp. 131-157). Amsterdam: IISG. Knippenberg, H. (2005). The Netherlands. Selling churches and building mosques. In H. Knippenberg (red.), The changing religious landscape of Europe (pp. 88-106). Amsterdam: Het Spinhuis. Knippenberg, H. & S. de Vos (1989). Spatial structural effects on Dutch church atten-
dance. Tijdschrift voor Economische en Sociale Geografie, 80, 164-170. Knippenberg, H. & S. de Vos (1991). Hedendaags kerkbezoek: de invloed van omge-
vingsfactoren en individuele kenmerken. Nederlands Theologisch Tijdschrift, 45,
46-59. Knippenberg, H. & H. van der Wusten, H (2001). De zuilen, hun lokale manisfesta-
ties en hun restanten in vergelijkend perspectief. In C. van Eijl, L. Heerma van
Voss & P. de Rooy (red.), Sociaal Nederland: contouren van de twintigste eeuw (pp. 129-150). Amsterdam: Aksant. hans knippenberg en sjoerd de vos Kok, J.A. de (1964). Nederland op de breuklijn Rome-Reformatie. Assen: Van Gorcum. Kruijt, J.P. (1928). De bevolking der Zaanstreek. Mens & Maatschappij, 4, 221-232 en
306-322. 125 Kruijt, J.P. (1933). De onkerkelijkheid in Nederland. Groningen: Noordhoff. Kruijt, J.P. (1935). Kerkelijkheid en onkerkelijkheid in Nederland. Socialistische
Gids, 20, 323-339 en 426-451. Kuiper, G. (1953). Beroep en kerkgenootschap. Mens & Maatschappij, 28, 25-41 en
85-99. Leeuwen, M.H.D. van & I. Maas. (2005). A short note on HISCLASS. http://historyof-
work.iisg.nl/docs/hisclass-brief.doc. Lijphart, A. (1968). The politics of accommodation: pluralism and democracy in the
Netherlands. Berkeley: University of California Press. Lijphart, A. (1977). Democracy in plural societies. New Haven/London: Yale University
Press. Maas, I., P.S. Lambert, R.L. Zijdeman, K. Prandy & M.H.D. van Leeuwen (2006). HIS-
CAM. The derivation then implementation of an historical occupational stratification
scale. Paper presented to the Sixth European Social Science History Conferen-
ce, RAI, Amsterdam. Mönnich, C.W. (1988). Ferdinand Domela Nieuwenhuis, theologie en anti-theo-
logie. In J. Frieswijk e.a. (red.), Ferdinand Domela Nieuwenhuis. De Apostel van de
Friese arbeiders. (pp. 63-79). Drachten/Leeuwarden: Friese Pers Boekerij. Need, A. & N.D. de Graaf (2005). Zich bekeren en wisselen van kerkgenootschap
in Nederland. Mens & Maatschappij, 80, 288-304. Norris, P. & R. Inglehart (2004). Sacred and Secular. Religion and politics worldwide. Cambridge: Cambridge University Press. Literatuur Prandy, K. (2000). The social interaction approach to the measurement and ana-
lysis of social stratification. International Journal of Sociology and Social Policy, 19,
215–249. Rasker, A.J. (1986). De Nederlandse Hervormde Kerk vanaf 1795. Kampen: Kok (derde
druk). Righart, H. (1986). De katholieke zuil in Europa. Meppel: Boom. Romein, J. & A. Romein (1973; oorspronkelijk 1938-1940). Erflaters van onze bescha-
ving. Amsterdam: Querido (tiende druk). Rooden, P. van (1996). Religieuze regimes. Over godsdienst en maatschappij in Neder-
land, 1570-1990. Amsterdam: Bert Bakker. Rooy, P. de (1995). Zes studies over verzuiling. Bijdragen en Mededelingen betreffende
de Gechiedenis der Nederlanden, 110, 380-392. Schepens, Th. (1991). Kerk in Nederland. Een landelijk onderzoek naar kerkbetrokken-
heid en kerkverlating. Tilburg: Tilburg University Press. Sengers, E. (red.) (2005). The Dutch and their Gods. Secularization and transformation of
religion in the Netherlands since 1950. Hilversum: Verloren. Stark, R. (1999). Secularization, R.I.P. Sociology of Religion, 60, 249-273. Stark, R. & L.R. Iannacone (1994). A supply-side reinterpretation of the ‘secualari-
zation’ of Europe. Journal for the Scientific Study of Religion, 33, 230-252. vroege ontkerkelijking in nederland vroege ontkerkelijking in nederland Staverman, M. (1954). Buitenkerkelijkheid in Friesland. Assen: Van Gorcum. Vaus, D.A. de (1982). The impact of children on sex related differences in church
attendance. Sociological Analysis, 43, 145-154. 126 Vaus, D.A. de (1984). Workforce participation and sex differences in church atten-
dance. Review of Religious Research, 25, 247-256. Vaus, D.A. de & I. McAllister (1987). Gender differences in religion: a test of the
structural location theory. American Sociological Review, 52, 472-481. Vaus, D.A. de & I. McAllister (1987). Gender differences in religion: a test of the
structural location theory. American Sociological Review, 52, 472-481. Verweij, J. (1998). Secularisering tussen feit en fictie. Tilburg: Tilburg University
Press. Vos, S. de (1997). De omgeving telt. Compositionele effecten in de sociale geografie. Proef-
schrift Universiteit van Amsterdam. Wunder, E. (2005). Religion in der postkonfessionellen Gesellschaft. München: Franz
Steiner Verlag. Functioneel analfabetisme in Nederland,
1775-1900 Onno Boonstra Anderhalf miljoen analfabeten in Nederland j
De media maken met enige regelmaat melding van een groot probleem in Neder-
land: er zijn zoveel analfabeten in ons land. Nu eens wordt beweerd dat het om
één, dan weer om twee, en soms zelfs om drie miljoen analfabeten gaat. Ieder
aantal is goed, zolang het ons doet duizelen. Want hoe kan het dat er zoveel
analfabeten wonen in een land waar de leerplicht al meer dan een eeuw geleden
is ingevoerd? Iedereen zal zo langzamerhand toch wel kunnen lezen en schrij-
ven? Het antwoord is eenvoudig. Natuurlijk kan vrijwel iedereen lezen en schrij-
ven. Maar het begrip analfabetisme is opgerekt. Het gaat nu om ‘functioneel
analfabetisme’ of ‘laaggeletterdheid’: de huidige analfabeet is iemand die onvol-
doende lees-, schrijf- en rekenvaardigheden bezit om in de moderne samenle-
ving te functioneren. Deze definitie is niet erg helder. Wat ‘onvoldoende’ is, wordt niet duidelijk
gemaakt, en dat geldt ook voor het begrip ‘functioneren in de moderne samen-
leving’. Dat heeft de stichting Lezen & Schrijven er niet van weerhouden om een
onderzoek naar ‘laaggeletterdheid’ in onze samenleving te entameren. In sep-
tember 2004 werden de uitkomsten van het onderzoek gepresenteerd. Volgens
het persbericht telde Nederland in dat jaar naar schatting 1,5 miljoen mensen (6% van de beroepsbevolking), die over
onvoldoende basisvaardigheden als lezen, schrijven en rekenen beschik-
ken, om volledig te kunnen functioneren in de samenleving. Volgens
het onderzoek is dit aantal groter (naar schatting 2,5 miljoen) omdat de
kenniseconomie naast deze basisvaardigheden ook andere competenties
vergt van werknemers. De gevolgen voor deze groep laaggeletterden is dan
ook aanzienlijk en vereist extra aandacht en samenwerking van overheid,
bedrijfsleven en burgers. (Lezen en Schrijven, 2004) naar schatting 1,5 miljoen mensen (6% van de beroepsbevolking), die over
onvoldoende basisvaardigheden als lezen, schrijven en rekenen beschik-
ken, om volledig te kunnen functioneren in de samenleving. Volgens
het onderzoek is dit aantal groter (naar schatting 2,5 miljoen) omdat de
kenniseconomie naast deze basisvaardigheden ook andere competenties
vergt van werknemers. De gevolgen voor deze groep laaggeletterden is dan
ook aanzienlijk en vereist extra aandacht en samenwerking van overheid,
bedrijfsleven en burgers. (Lezen en Schrijven, 2004) Met drie taalfouten (en minstens één rekenfout) in één alinea bewijst de stich-
ting inderdaad dat niet iedereen over basisvaardigheden als lezen, schrijven en
rekenen beschikt, en tegelijkertijd ook dat zoiets kennelijk geen hinderpaal is
om te functioneren in onze samenleving. Anderhalf miljoen analfabeten in Nederland 128 Daarmee maakt de stichting zelf al onbedoeld duidelijk hoe lastig het is om
een eenduidige definitie van het begrip ‘functioneel analfabetisme’ te geven. Wie is functioneel analfabeet? Is het degene die de gebruiksaanwijzing van zijn
dvd-speler niet begrijpt, is het degene die zich in de luren laat leggen door ‘Haags
taalgebruik’, is het degene die vergeefs het sms’je van zijn dochter probeert te
lezen, of is het degene die, net als de stichting Lezen & Schrijven, met cijfers goo-
chelt zonder te beseffen wat de betekenis van die cijfers is? Analfabetisme in de negentiende eeuw Het idee dat analfabetisme mensen ernstig hindert om goed te functioneren in
de samenleving, en het idee dat de kenniseconomie vereist dat de leden van de
samenleving goed kunnen lezen en schrijven, is onomstreden, en dat al sinds
twee eeuwen. Dat lijkt terecht, want zeker op individueel niveau zijn prachtige
voorbeelden te vinden van mensen die dankzij het verwerven van lees- en schrijf-
vaardigheden vooruit zijn gekomen in de maatschappij. Dat blijkt bijvoorbeeld
uit de autobiografie die omstreeks 1890 door de toen 75-jarige Pieter Arkenbout
is geschreven.1 Hij vertelt daarin onder meer over wat hem overkwam toen hij in
1837 als soldaat in Bergen op Zoom gelegerd was. Tijdens het schrijven van een
brief aan zijn ouders werd hij gestoord door een soldaat, J. van der Kolk gehe-
ten. Toen ik daarmede bezig was, hinderde het mij zeer dat een oudere soldaat,
onafgebroken achter mij daarna stond te kijken, net zoo lang tot ik eene
bladzijde geschreven had, en bij het omvouwen hem aankeek, waarop de
man met tranen in de oogen zeide “ik zou er mijn pink wel voor willen
missen wanneer ik dat ook zoo kon” dit gezegde veranderde terstond mijn
gemoedstoestand, en was ik eerst met haat tegen hem vervuld om zijn vrij-
postigheid; nu gevoelde ik medelijden met hem, wat nog toenam, toen hij
mij mededeelde niet alleen niet te kunnen lezen, maar zelfs geen enkele
letter te kennen, maar het nog zoo gaarne zou leeren. Arkenbout en Van der Kolk maakten een afspraak. In ruil voor de dagelijkse
poetsbeurt van de patroontas en de bajonetschede, zou Pieter de oude soldaat
leren lezen en schrijven. Zo gezegd, zo gedaan. Maar na de diensttijd scheidden
zich beider wegen. Soldaat Van der Kolk ging terug naar Goidschalxoord, zijn
geboortedorp in de Hoekse Waard. Ruim 25 jaar ging voorbij tot Arkenbout het
volgende overkwam. onno boonstra Op zekeren avond in de maand Juny (1864) van een bezoek aan mijn broe-
der, die een klein uurtje van de stad woonde, terugkeerende, ontmoette ik
bij eene boerderij een vlasboer, die evenals ik naar de stad ging om er te
logeeren met wien ik al spoedig in gesprek was over een en ander en hem
vroeg van waar hij kwam, waarop hij zeide “Goidschalkoord”, en vroeg
hem toen of hij J. van der Kolk ook kende en zijne omstandigheden hem
bekend waren wat hij met ja! Analfabetisme in de negentiende eeuw beantwoordde en niet alleen ik, maar zelfs
het kleinste kind op het dorp kent hem want hij is al van het eerste jaar af
dat hij van de dienst gekomen is, veldwachter, gemeente- en polderbode
enz. en begon toen de zamenloop van omstandigheden te verhalen die
tot zijn benoeming geleid hadden want daar hij de eenige sollicitant uit
de gemeente was had hij anders wel kans, maar alle raadsleden waaron-
der hij toen ook pas behoorde wisten dat hij nimmer school had gegaan
en konden dus niet gelooven dat hij zijn verzoekschrift zelf geschreven
had vóór dat men hem van zijn werk liet halen en hij daar in de raadka-
mer voor aller oog geschreven had waarop hij met algemeene stemmen
benoemd werd. (Arkenbout, z.j.) 129 Op macroniveau is de relatie tussen economie en onderwijs minder eenduidig. Halverwege de twintigste eeuw vonden sociologen in negentiende-eeuwse Euro-
pese cijfers duidelijke aanwijzingen dat landen met een hoge onderwijsdeelna-
me veel sneller economisch waren gemoderniseerd dan landen waar dat niet
het geval was geweest, en in twintigste-eeuwse cijfers uit andere werelddelen
dat daar, waar de onderwijsdeelname sterk groeide, de economische ontwikke-
ling eveneens een sterk stijgende lijn vertoonde (Sanderson, 1972; Inkeles, 1973;
Cipolla, 1967). Hun conclusie was dan ook dat investeringen in onderwijs in derde-
wereldlanden zich zouden uitbetalen in een forse economische groei. Later werd
duidelijk dat er van een direct causaal verband tussen onderwijs en economische
groei geen sprake is (Gylafson 2001; Wolf 2002), en dat voor Europa in de negen-
tiende eeuw slechts bij uitzondering een samenhang aangetoond kon worden:
in de negentiende eeuw begon de economische groei van veel landen met een
hoog onderwijsniveau (zoals Nederland en Zweden) bijvoorbeeld een stuk later
dan die van landen met een lager onderwijsniveau (zoals Engeland of België). Ook op macroniveau bleken kanttekeningen te plaatsen bij het idee dat anal-
fabetisme mensen hindert om goed te functioneren in de samenleving. De Ame-
rikaan Harvey J. Graff heeft bijvoorbeeld een groot deel van zijn werkzame leven
besteed aan pogingen om het ongelijk van die veronderstelling aan te tonen
(Graff, 1979, 1981, 1987). Belangrijkste argument van Graff is dat de industriële
modernisering van Europa in het midden van de negentiende eeuw helemaal
niet op hoger opgeleide arbeidskrachten zat te wachten, en dat de landarbeiders
en de werkers in de fabrieken zelf ook helemaal geen boodschap hadden aan de
vaardigheid om te kunnen lezen en schrijven. Analfabetisme in de negentiende eeuw De Economist van 1852 meende bij-
voorbeeld het volgende. functioneel analfabetisme in nederland, 1775-1900 Er zijn sommige mensen, vooral bij de boerenstand, die niet veel op hebben
met de scholen. – Niet zelden zijn het diegenen die vroeger zelf niet school
gingen, en die alzoo meenen, daarin een sprekend voorbeeld te zien, dat
men het al zeer ver kan brengen zonder dat. (Wenken, 1852) 130 130 En volgens de Fransman Nihaud (geb. 1815) werd zijn vader door zijn grootvader
met de volgende woorden gekapitteld, omdat hij zijn zoon naar school wilde
sturen. Ni mes frères, ni toi, ni moi, n’avons jamais appris à connaître nos lettres
et nous avons mangé du pain tout de même. 2 Dit waren echter woorden van een minderheid. Het was halverwege de nege
tiende eeuw in Nederland – in alle regio’s, in alle klassen en in alle gezindt tiende eeuw in Nederland – in alle regio’s, in alle klassen en in alle gezindten
– de gewoonste zaak van de wereld geworden om kinderen onderwijs te laten
volgen (Knippenberg, 1986). Kennelijk vormden de kosten die schoolgaan met
zich meebracht zelfs voor arme ouders geen beletsel, net zo min als katholieke
ouders er bezwaar tegen hadden dat hun kinderen onderwijs volgden dat op
protestantse leest was geschoeid. Dat wekt de suggestie dat, ook al zou analfa-
betisme objectief gezien geen belemmering vormen bij het uitoefenen van een
beroep, de ouders desondanks de gedachte koesterden dat de vaardigheid om
te lezen en schrijven hun kinderen wél verder zou helpen in de maatschappij. Velen prefereerden de investering in onderwijs boven direct gewin, omdat ze
voor hun kinderen er bepaalde voordelen van verwachtten: kunnen lezen en
schrijven was misschien niet per se noodzakelijk, maar het gaf een zeker gemak
bij de uitoefening van het beroep en wellicht ook betere kansen op een mooie
beroepsloopbaan (Boonstra, 1993). Een onderzoek naar analfabetisme en beroepsmobiliteit Een onderzoek naar analfabetisme en beroepsmobiliteit
Het is dus maar de vraag of onderwijs kinderen wel het economische voordeel
bracht dat hun ouders ervan verwachtten. Deze vraag kan beantwoord worden
door na te gaan of analfabeten in hun beroepsloopbaan gehinderd werden door
de handicap niet te kunnen lezen en schrijven. Daarbij wordt zowel gekeken
naar intra- als intergenerationele beroepsmobiliteit. ‘Beroepsmobiliteit’ wordt
geoperationaliseerd als het stijgen of dalen op de sociale ladder op basis van een
verandering van beroepsklasse. Vanwege het gebrek aan bruikbare beroepsdata
over vrouwen blijft het onderzoek beperkt tot mannen. De beroepen zijn geco-
deerd volgens het classificatieschema van HISCO (Van Leeuwen, Maas & Miles,
2002), waarna ze in beroepsklassen zijn gegroepeerd volgens de HISCLASS-inde-
ling (Maas & Van Leeuwen, 2004).3 onno boonstra De HISCLASS-indeling kent 12 klassen, die ten behoeve van dit onderzoek
zijn ingeperkt tot zes. Klasse 1
hoger
1
Hogere managers
2
Hogere vrije beroepen
3
Lagere managers
Klasse 2
middelbaar
4
Lagere vrije beroepen, klerken en winkelpersoneel
5
Lagere klerken en winkelpersoneel
Klasse 3
geschoold
6
Voorlieden
7
Geschoolde arbeiders
Klasse 4
boeren
8
Boeren
Klasse 5
laaggeschoold
9
Laaggeschoolde arbeiders
10 Laaggeschoolde boerenarbeiders
Klasse 6
ongeschoold
11 Ongeschoolde arbeiders
12 Ongeschoolde boerenarbeiders 131 131 Wanneer iemand naar een klasse gaat met een lager nummer is er dus sprake van
opwaartse mobiliteit; iemand die overstapt naar een klasse met een hoger num-
mer is neerwaarts mobiel. De kansen op beroepsmobiliteit worden normaliter
gezien als een resultante van ‘achievement’, de eigen verdienste, en ‘ascription’,
het sociaal en cultureel kapitaal dat door afkomst is verworven (Blau & Duncan,
1967). ‘Achievement’ is in dit onderzoek vanzelfsprekend geoperationaliseerd als
‘alfabetisme’, terwijl ‘ascription’ op twee manieren is gedefinieerd: als sociaal
kapitaal is de beroepsklasse van de vader gemeten, en als cultureel kapitaal het
alfabetisme van de vader. Dat betekent dat dus niet alleen gekeken is naar de
beroepspositie van de vader ter beoordeling van de loopbaan van de zoon, maar
ook of de zoon uit een alfabeet of analfabeet gezin afkomstig is. Analfabetisme is op een eenvoudige manier gedefinieerd. Een respondent
wordt als analfabeet beschouwd wanneer hij niet in staat is om een handteke-
ning te zetten op plekken in de ambtelijke registers waar hij geacht werd dit
wel te doen. De handtekening is een goede maat om alfabeten van analfabeten
te scheiden. Een onderzoek naar analfabetisme en beroepsmobiliteit Het zetten van een handtekening is een vorm van schrijven, en in
de negentiende eeuw werd op school pas schrijven geleerd nadat de leerlingen
eerst hadden leren lezen. Iemand die kan schrijven kan dus ook lezen en is dus
alfabeet. Het is ook een bruikbare maat, omdat de handtekening in veel massa-
bronnen opduikt. Huwelijksregisters bevatten bijvoorbeeld een handtekening
van bruid en bruidegom, alsmede van hun ouders en eventuele getuigen. En ook
wanneer vaders de geboorte van een kind komen aangeven, dient een handteke-
ning in het geboorteregister te worden geplaatst. De HSN, de Historische Steekproef Nederlandse bevolking, vormt een
redelijk goede bron voor dit onderzoek, zeker omdat in dit geval niet alleen
gebruik is gemaakt van data uit de bevolkingsregisters (HSN-dataset Levenslopen
release 2007.01), maar ook van de akten (HSN-dataset Akten burgerlijke stand, release functioneel analfabetisme in nederland, 1775-1900 2007.01). Uit de huwelijksakten kan worden afgeleid of een onderzoekspersoon,
zijn huwelijkspartner, alsmede zijn ouders en schoonouders, een handtekening
konden zetten. Daarnaast staan hun beroepen in de huwelijksakten vermeld. De
handtekening en het beroep zijn ook te vinden in de geboorteakten, terwijl in
de bevolkingsregisters en op persoonskaarten eveneens beroepen vermeld staan,
zodat ook deze bronnen in het onderzoek zijn betrokken. 132 Op dit moment zijn alle geboorteakten, maar nog niet alle huwelijksakten
verzameld en ondergebracht in de dataset met de akten. Ook gegevens uit de
bevolkingsregisters zijn nog niet volledig ingevoerd (zie de bijdrage van Mande-
makers in dit boek). Alle huwelijksakten van vóór 1853 ontbreken bijvoorbeeld,
en de huwelijksakten uit de periode 1853-1883 zijn voor slechts een beperkt aan-
tal regio’s beschikbaar. Dat maakt het gebruik van de HSN voor dit onderzoek
naar alfabetisme problematisch: juist in de eerste helft van de negentiende eeuw
is analfabetisme nog een vrij normaal verschijnsel; het aantal mensen dat na
1850 is geboren en analfabeet is gebleven is daarentegen bijzonder gering. Om
die laatste reden is besloten om het onderzoek te beperken tot mannen die voor
het jaar 1900 zijn geboren. Een tweede probleem betreft de representativiteit van de steekproef. Van
alleen gehuwde onderzoekspersonen weten we of ze hun handtekening hebben
gezet; ongehuwden zijn niet in de analyse opgenomen. Ook de omstandigheid
dat in de HSN tot nu toe vooral huwelijksakten en gegevens uit bevolkingsregis-
ters zijn verzameld van personen die in de directe omgeving van de plaats van
huwen zijn geboren, zorgt voor een vertekening. Een onderzoek naar analfabetisme en beroepsmobiliteit Omdat het aantal onderzoekspersonen anders te klein zou zijn, én omdat
de representativiteit toch al te wensen over laat, is besloten om een nieuwe
onderzoekspopulatie te creëren die niet alleen bestaat uit officiële mannelijke
HSN-onderzoekspersonen, maar ook uit andere mannen die in de HSN-releases
opduiken: de partners van vrouwelijke HSN-onderzoekspersonen bijvoorbeeld,
of de vaders van de bruiden en bruidegoms uit de huwelijksakten. De potentiële
onderzoekspopulatie komt daarmee op maar liefst 107.552 personen. In werke-
lijkheid is het aantal respondenten een stuk kleiner. Van sommige mannen ont-
breekt een beroepsvermelding of is het vermelde beroep niet te classificeren;
voor weer anderen geldt dat ze na 1900 en dus buiten de onderzoeksperiode zijn
geboren. Het aantal respondenten loopt daardoor terug tot 31.274. Zwaarwegen-
der is dat voor het onderzoek naar intragenerationele mobiliteit alleen respon-
denten zijn geselecteerd van wie twee of meer beroepsvermeldingen bekend zijn
en van wie de eerste en laatste vermelding minstens een jaar uit elkaar liggen,
en dat voor het onderzoek naar intergenerationele mobiliteit alleen HSN-onder-
zoekspersonen zijn geselecteerd van wie ook beroepsgegevens van hun vader
bekend zijn. Het aantal respondenten loopt daardoor terug tot 10.357 voor wat
betreft het onderzoek naar intragenerationele, en 9.457 voor wat betreft het
onderzoek naar intergenerationele mobiliteit.4 onno boonstra Alfabeten, analfabeten en hun beroepsloopbaan Zoals hierboven reeds is aangegeven, was analfabetisme in Nederland een feno-
meen dat in de loop van de negentiende eeuw uit de samenleving verdween. Figuur 1 staaft deze bewering. Rond 1800 ging een ruime meerderheid van de
jongens en de helft van de meisjes naar school, en er is reden om te vermoe-
den dat die niveaus ook in de twee eeuwen daarvoor gehaald werden (Boonstra,
2009). In de negentiende eeuw liep de onderwijsdeelname verder op, en begon-
nen de analfabetismecijfers verder te dalen. 133 Van alle jongens die in het laatste kwart van de achttiende eeuw werden
geboren leerde ongeveer 75 procent lezen en schrijven, en wist een minderheid
van 25 procent zich niet in deze vaardigheden te bekwamen. Vanaf dat moment
vond er een langzame, maar gestage daling van dat percentage plaats. Rond 1800
was het 20 procent, rond 1820 15 procent, in 1830 10 procent, en in 1860 5 pro- Figuur 1 Ontwikkeling van het analfabetisme in Nederland naar geboortejaar (1775-1900),
mannen en vrouwen Bron: HSN-dataset Akten, release 2007.01. Voor mannen: geboorteakten (vaders van HSN-onderzoeks-
personen) en huwelijksakten (mannelijke onderzoekspersonen en echtgenoten van vrouwelijke onder-
zoekspersonen). Voor vrouwen: huwelijksakten (vrouwelijke onderzoekspersonen en echtgenoten van
mannelijke onderzoekspersonen). N mannen = 107.552, N vrouwen = 45.794. Bron: HSN-dataset Akten, release 2007.01. Voor mannen: geboorteakten (vaders van HSN-onderzoeks-
personen) en huwelijksakten (mannelijke onderzoekspersonen en echtgenoten van vrouwelijke onder-
zoekspersonen). Voor vrouwen: huwelijksakten (vrouwelijke onderzoekspersonen en echtgenoten van
mannelijke onderzoekspersonen). N mannen = 107.552, N vrouwen = 45.794. Bron: HSN-dataset Akten, release 2007.01. Voor mannen: geboorteakten (vaders van HSN-onderzoeks-
personen) en huwelijksakten (mannelijke onderzoekspersonen en echtgenoten van vrouwelijke onder-
zoekspersonen). Voor vrouwen: huwelijksakten (vrouwelijke onderzoekspersonen en echtgenoten van
mannelijke onderzoekspersonen). N mannen = 107.552, N vrouwen = 45.794. functioneel analfabetisme in nederland, 1775-1900 functioneel analfabetisme in nederland, 1775-1900 cent. Rond 1880 wordt het nulpunt bereikt. Deze daling deed zich niet alleen bij
jongens voor: ook de meisjes die rond 1880 werden geboren, leerden allemaal
lezen en schrijven. De analfabetismedaling onder meisjes verliep dus specta-
culairder, want het analfabetismeniveau lag in het eerste kwart van de negen-
tiende eeuw nog rond de 50 procent. Het percentage van 25 procent werd door
meisjes rond 1840 bereikt, 20 procent omstreeks 1850, 10 procent rond 1860, en
5 procent omstreeks 1875. 134 De daling van het analfabetisme was een fenomeen dat zich in het gehele
land voordeed. Alfabeten, analfabeten en hun beroepsloopbaan Er waren echter regio’s waar de percentages hoger lagen dan
elders en waar de daling dientengevolge ook wat vertraagd plaats vond. Figuur 2
toont de regionale verschillen in analfabetismeniveaus voor het geboortecohort
1800-1825. Daarnaast was analfabetisme een verschijnsel dat zich grotendeels tot de aarnaast was analfabetisme een verschijnsel dat zich grotendeels tot de Daarnaast was analfabetisme een verschijnsel dat zich grotendeels tot de Figuur 2 Percentage analfabete mannen per gemeente in geboortecohort 1800-1825 Ontleend aan Boonstra (2009). N = 21.334. < 10%
10–20%
20–30%
> 30%
< 10%
10–20%
20–30% Ontleend aan Boonstra (2009). N = 21.334. < 1
10–2 onno boonstra lagere klassen beperkte. Figuur 3 laat zien dat de lagere ambtenaren, de geschool-
de arbeiders en de boeren (beroepsklassen 2 t/m 4), die in de peropde 1775-1800
waren geboren, analfabetismeniveaus kenden tussen de 10 en 20 procent, terwijl
de laag- en ongeschoolde arbeiders (klassen 5 en 6) boven de 20 procent uitkwa-
men. Vijftig jaar later, in de periode 1825-1850, zijn die laatste twee klassen de
enige waar meer dan 10 procent niet leerde lezen en schrijven; nog eens vijftig
jaar later is analfabetisme volledig uit de Nederlandse geboortecohorten verdwe-
nen. 135 De negentiende eeuw was een eeuw van modernisering in landbouw en
industrie. Fabrieken en werkplaatsen verrezen in het hele land, en de vraag om
arbeiders groeide snel. Vooral het aantal industriearbeiders nam toe. Dat blijkt
uit de cijfers: het percentage fabrieksarbeiders groeide van 4 procent in 1850
tot 15 procent vijftig jaar later, terwijl het percentage boeren en ambachtslie-
den in diezelfde periode daalde van 34 tot 20 procent (Mandemakers, 2001). Het
gevolg is dat de negentiende eeuw vooral een neerwaartse mobiliteit laat zien:
het aantal personen dat gedurende hun beroepsloopbaan een of meer klassen
daalde is groter dan het aantal personen dat steeg. Neerwaartse mobiliteit was
dus omvangrijker dan opwaartse mobiliteit, maar in de loop van de eeuw nam
de neerwaartse mobiliteit wel af: van 49 procent in 1775-1800 tot 21 procent in
1875-1900. De opwaartse mobiliteit daarentegen werd gedurende de eeuw lang-
zaam groter: bedroeg het percentage in 1775-1800 11 procent; honderd jaar later
was dat gestegen tot 14 procent. Figuur 3 Percentage analfabete mannen per beroepsklasse naar geboortejaar (1775-1925) Figuur 3 Percentage analfabete mannen per beroepsklasse naar geboortejaar (1775-1925)
Bron: HSN-datasets Akten en Levenslopen, release 2007.01; N = 31.274. Alfabeten, analfabeten en hun beroepsloopbaan 0
5
10
15
20
25
30
35
1775-1800
1800-1825
1825-1850
1850-1875
1875-1900
1900-1925
% analfabeet
geboortejaar
klasse 1
klasse 2
klasse 3
klasse 4
klasse 5
klasse 6 0
1775-1800
1800-1825
1825-1850
1850-1875
1875-1900
1900-1925
geboortejaar Bron: HSN-datasets Akten en Levenslopen, release 2007.01; N = 31.274. functioneel analfabetisme in nederland, 1775-1900 Figuur 4 Intragenerationele beroepsmobiliteit van alfabeten en analfabeten naar geboorte-
jaar (1775-1900)
Bron: HSN-datasets Akten en Levenslopen, release 2007.01; N = 10.496. 0
10
20
30
40
50
60
1775-1800
1800-1825
1825-1850
1850-1875
1875-1900
geboortejaar
% mobiel
analfabeet opwaarts
analfabeet neerwaarts
alfabeet opwaarts
alfabeet neerwaarts Figuur 4 Intragenerationele beroepsmobiliteit van alfabeten en analfabeten naar geboorte-
jaar (1775-1900) Figuur 4 Intragenerationele beroepsmobiliteit van alfabeten en analfabeten naar geboorte-
jaar (1775-1900) analfabeet opwaarts
analfabeet neerwaarts
alfabeet opwaarts
alfabeet neerwaarts Bron: HSN-datasets Akten en Levenslopen, release 2007.01; N = 10.496. Figuur 4 geeft zowel de mobiliteit van alfabeten als van analfabeten. Beide
groepen ontwikkelden zich tot op zekere hoogte langs gelijke lijnen, zij het dat
de neerwaartse mobiliteit van analfabeten gedurende de eeuw niet afnam, en de
opwaartse mobiliteit niet toenam. Een ander verschil is dat de opwaartse mobili-
teit van analfabeten gemiddeld hoger was dan die van alfabeten, en, omgekeerd,
de neerwaartse mobiliteit van alfabeten hoger was dan die van analfabeten. Deze cijfers zijn echter bedrieglijk. Voor een deel is de grotere opwaartse en
de geringere neerwaartse mobiliteit van analfabeten het gevolg van de beroeps-
klasse waarin ze verkeerden: wanneer alle analfabeten in de laagste klasse zou-
den vertoeven, zou er zelfs helemaal geen kans op neerwaartse mobiliteit meer
zijn. Om de effecten van analfabetisme daadwerkelijk in kaart te brengen zal het
effect in samenhang met andere variabelen moeten worden onderzocht. Daar-
toe is een multivariate analyse gemaakt van de kans op opwaartse of neerwaart-
se mobiliteit gedurende de beroepsloopbaan van een groep die bestaat uit (1)
mannelijke onderzoekspersonen van de HSN, (2) hun vaders, (3) echtgenoten van
vrouwelijke onderzoekspersonen en (4) hun vaders. Als analysetechniek is een
binomiale logistische regressie gekozen, waarin degenen die mobiel waren zijn
afgezet tegen degenen die niet van beroepsklasse veranderden. Als startmodel is
een model gekozen waarin drie onafhankelijke variabelen zijn opgenomen: het
analfabetisme van de respondent, de periode waarin de respondent geboren is,
de beroepsklasse van waaruit de mobiliteit is begonnen, alsmede alle tweeweg
interactie-effecten tussen deze variabelen. Alfabeten, analfabeten en hun beroepsloopbaan Door middel van een methode van
‘stepwise backward elimination’ zijn alle interactie-effecten die niet significant onno boonstra bleken, stapsgewijze uit het model verwijderd; de hoofdeffecten zijn, ook al ble-
ken ze niet significant te zijn, in het model blijven staan. In de analyse zijn alleen respondenten toegelaten waarvan de eerste beroeps-
vermelding minstens een jaar ouder is dan de laatste vermelding. De beroepsver-
meldingen zijn ondergebracht in de eerder vermelde zes beroepsklassen. Tevens
zijn in de analyse van opwaartse mobiliteit degenen die al in het begin van hun
beroepsloopbaan in klasse 1 vertoefden en dus onmogelijk verder konden stij-
gen, weggelaten; uit de analyse van neerwaartse mobiliteit zijn degenen die in
de laagste klasse startten, verwijderd. In tabel 1 zijn de belangrijkste resultaten
samengevat. 137 Tabel 1 laat zien dat de kans op opwaartse mobiliteit gedurende de loop-
baan van de respondent werd bepaald door twee factoren: geboorteperiode en
beroepsklasse. Wie aan het einde van de achttiende eeuw werd geboren, had
een grotere kans om te stijgen dan degene die in de loop van de negentiende
eeuw werd geboren. De kans om te stijgen werd echter gedempt door de beroeps-
klasse waarin de respondent zijn carrière startte: wie in een hogere klasse begon,
werd gehinderd om op de beroepsladder te stijgen; wie in een lage klasse begon,
liep daarentegen tegen minder hindernissen aan. Dat laatste is vanzelfsprekend
geen opzienbarende uitkomst. Interessanter is om te zien dat er een significant
interactie-effect bestond tussen beroepsklasse en geboorteperiode. Aan het einde
van de achttiende en het begin van de negentiende eeuw waren de kansen om te
stijgen bijvoorbeeld voor laaggeschoolde mannen toch wel een stuk kleiner dan
de hoofdeffecten suggereren. Maar de meest opvallende uitkomst is dat analfa-
betisme kennelijk geen rol speelde: of iemand kon lezen of schrijven had geen
invloed op zijn kansen om te stijgen op de beroepsladder. Bij neerwaartse intragenerationele mobiliteit blijken dezelfde variabelen
een rol te spelen. Geboorteperiode en beroepsklasse tonen ook in dit geval sig-
nificante effecten. In de loop van de negentiende eeuw lijkt de kans op neer-
waartse mobiliteit toe te nemen, en hadden degenen die in de hoogste klassen
bivakkeerden vanzelfsprekend meer kans om te dalen dan degenen die in een
lage klasse startten. Alfabeten, analfabeten en hun beroepsloopbaan Het significante interactie-effect dat er tussen beide variabe-
len bestaat zorgt er echter voor dat de toenemende kansen op neerwaartse mobi-
liteit gedurende de negentiende eeuw voor de meeste beroepsklassen werden
gedempt. Analfabetisme heeft opnieuw geen significant eigen effect: wie niet
kon lezen en schrijven had niet meer kans om op de beroepsladder te dalen dan
alfabeten. Beide analyses maken duidelijk dat de kans op opwaartse of neerwaartse
mobiliteit door twee van de drie in het model ingebrachte variabelen werd beïn-
vloed: geboorteperiode en beroepsklasse. De kans op opwaartse mobiliteit nam
in de loop van de tijd af, en die op neerwaartse mobiliteit toe. Ook oefende het
beroep van de respondent een belangrijk hoofdeffect uit: wie tot de laagste klas-
sen behoorde had duidelijk meer kans om te stijgen op de beroepsladder en wie
tot de hoogste klassen behoorde had duidelijk meer kans om te dalen. onno boonstra Alfabeten, analfabeten en hun beroepsloopbaan Die kan- functioneel analfabetisme in nederland, 1775-1900 functioneel analfabetisme in nederland, 1775-1900 Tabel 1 Logistische-regressieanalyse van opwaartse en neerwaartse intragenerationele mobi-
liteit Tabel 1 Logistische-regressieanalyse van opwaartse en neerwaartse intragenerationele mobi-
liteit liteit
Opwaarts mobiel
Neerwaarts mobiel
B
Exp(B)
B
Exp(B)
Intercept
-0,54**
-
-1,02**
-
Analfabetisme
-0,02
1,26
0,12
0,89
Analfabeet
0
-
0
-
Alfabeet
Geboortecohort
1775-1800
2,82**
16,73
-3,11**
0,05
1800-1825
1,87**
6,45
-2,51**
0,08
1825-1850
1,54**
4,66
-1,37**
0,25
1850-1875
0,48**
1,62
-0,63**
0,53
1875-1900
0
-
0
-
Beroepsklasse
Klasse 1
1,59**
4,90
Klasse 2
-2,23**
0,11
0,34*
1,41
Klasse 3
-2,00**
0,14
0,29*
1,34
Klasse 4
-2,92**
0,05
0,01
1,01
Klasse 5
-0,85**
0,43
0
-
Klasse 6
0
-
Geboortecohort * beroepsklasse
1775-1800 * Klasse 1
4,19**
66,11
1800-1825 * Klasse 1
3,70**
40,62
1825-1850 * Klasse 1
2,29**
9,85
1850-1875 * Klasse 1
1,02**
2,78
1875-1900 * Klasse 1
0
-
1775-1800 * Klasse 2
-2,98**
0,05
5,27**
195,00
1800-1825 * Klasse 2
-1,63**
0,20
4,23**
68,45
1825-1850 * Klasse 2
-1,28**
0,28
3,07**
21,56
1850-1875 * Klasse 2
-0,73
0,48
1,38**
3,97
1875-1900 * Klasse 2
0
-
0
-
1775-1800 * Klasse 3
-2,76**
0,06
4,74**
114,35
1800-1825 * Klasse 3
-2,10**
0,12
3,57**
35,67
1825-1850 * Klasse 3
-1,64**
0,20
2,13**
8,41
1850-1875 * Klasse 3
-0,61**
0,54
1,10**
3,02
1875-1900 * Klasse 3
0
-
0
-
1775-1800 * Klasse 4
-2,28**
0,10
5,11**
165,26
1800-1825 * Klasse 4
-1,29**
0,28
3,88**
48,56
1825-1850 * Klasse 4
-0,95*
0,39
2,37**
10,74
1850-1875 * Klasse 4
-0,26
0,77
0,99**
2,70
1875-1900 * Klasse 4
0
-
0
-
1775-1800 * Klasse 5
-4,79**
0,01
0
- 138 onno boonstra Opwaarts mobiel
Neerwaarts mobiel
B
Exp(B)
B
Exp(B)
1800-1825 * Klasse 5
-2,74**
0,06
0
-
1825-1850 * Klasse 5
-1,77**
0,17
0
-
1850-1875 * Klasse 5
-0,73**
0,48
0
-
1875-1900 * Klasse 5
0
-
0
-
1775-1800 * Klasse 6
0
-
1800-1825 * Klasse 6
0
-
1825-1850 * Klasse 6
0
-
1850-1875 * Klasse 6
0
-
1875-1900 * Klasse 6
0
-
N=6.845
N=8.325
* = p < 0,05; ** = p < 0,01. Bron: HSN datasets Akten en Levenslopen, release 2007.01. 139 Bron: HSN datasets Akten en Levenslopen, release 2007.01. Alfabeten, analfabeten en hun beroepsloopbaan sen waren wel afhankelijk van de tijd: wie in het begin van de eeuw tot de hoog-
ste klassen behoorde, had significant meer kans om te stijgen dan degenen die
aan het eind van de eeuw die klasse bevolkten; wie in de periode 1775-1800 tot de
lager geschoolde beroepsklasse behoorde had duidelijk meer kans om te dalen
op de beroepsladder dan degenen die in latere perioden tot die klasse behoor-
den. Het effect van analfabetisme is in beide analyses afwezig. De schattingen wij-
zen weliswaar in de te verwachten richting (analfabeten hebben minder kans op
opwaartse en meer kans op neerwaartse mobiliteit), maar de effecten zijn verre
van significant. De conclusie lijkt daarom gewettigd dat het voor je beroepsloop-
baan niet veel uitmaakte of je analfabeet was of niet. Analfabetisme en intergenerationele sociale mobiliteit Het mag dan zo zijn dat de vaardigheid om te kunnen lezen en schrijven niet
echt verder hielp in de beroepsloopbaan, dat wil nog niet zeggen dat men zich,
door wél te kunnen schrijven, niet zou kunnen onttrekken aan het ouderlijk
milieu. Om dat uit te zoeken is een analyse gemaakt van de intergenerationele
mobiliteit van alfabeten en analfabeten. In deze analyse vormen de mannelijke
HSN-onderzoekspersonen en de echtgenoten van vrouwelijke HSN-onderzoeks-
personen de onderzoekspopulatie. Van hen moesten hun alfabetisme en hun
beroep bekend zijn, alsmede het alfabetisme en beroep van hun vaders. Het aan-
tal respondenten komt daarmee op 9.457. Dat ook het alfabetisme van de vader in de analyse wordt betrokken, heeft
een reden. Het onttrekken aan je ouderlijk milieu is geen sinecure. Dat blijkt uit
het feit dat kinderen van analfabete ouders een heel grote kans hebben om zelf
analfabeet te blijven. De ‘overerving’ van analfabetisme wordt getoond in tabel 2. functioneel analfabetisme in nederland, 1775-1900 Kind
Zoon
Dochter
Vader
9,7
(N=32.332)
11,4
(N=15.578)
9,1
(N=16.754)
Moeder
14,3
(N=23.387)
12,8
(N=10.713)
15,4
(N=12.674)
Bron: HSN datasets Akten en Levenslopen, release 2007.01. 140 De log-odds laten zien dat kinderen van een analfabete vader, maar meer nog
kinderen van een analfabete moeder, een onevenredig grote kans hadden om
zelf ook analfabeet te blijven. De relatie was het sterkst tussen moeder en doch-
ter, en het minst sterk tussen vader en dochter. De logg-odds van tabel 2 mogen
dan fors zijn, ze zijn niet zo fors dat er geen mogelijkheden voor intergeneratio-
nele beroepsmobiliteit bestonden. Integendeel, van alle analfabete responden-
ten die bijvoorbeeld in het eerste kwart van de negentiende eeuw werden gebo-
ren, was 20 procent werkzaam in een beroepsklasse die hoger lag dan de klasse
waarin zijn vader werkzaam was. Voor alfabete respondenten was het percentage
opwaarts mobielen zelfs nog een stuk hoger. Zie figuur 5. Figuur 5 laat zien dat de kans op opwaartse mobiliteit gedurende de gehele
negentiende eeuw het grootst was voor mannen die net als hun vaders konden
lezen en schrijven. In het begin hadden alfabete zoons van analfabete vaders Figuur 5 Intergenerationele opwaartse beroepsmobiliteit naar alfabetisme van vader en zoon
en geboortejaar (1800-1900)
intergenerationele opwaartse beroepsmobiliteit van alfabeten en analfabeten Bron: HSN-datasets Akten en Levenslopen, release 2007.01; N = 9.457. Analfabetisme en intergenerationele sociale mobiliteit 0
5
10
15
20
25
30
35
40
45
1800-1825
1825-1850
1850-1875
1875-1900
geboortejaar
% mobiel
beiden analfabeet
vader analfabeet, zoon alfabeet
vader alfabeet, zoon analfabeet
beiden alfabeet 0
5
10
15
20
25
30
35
40
45
1800-1825
1825-1850
1850-1875
1875-1900
geboortejaar
% mobiel
beiden an
vader ana
vader alfa
beiden alf beiden analfabeet
vader analfabeet, zoon alfabeet
vader alfabeet, zoon analfabeet
beiden alfabeet Bron: HSN-datasets Akten en Levenslopen, release 2007.01; N = 9.457. onno boonstra dezelfde kansen, maar die lijken in de loop van de eeuw te dalen. Analfabete
zoons hadden minder kans, en dat geldt (althans na 1825) nog sterker wanneer
hun vaders ook niet konden lezen en schrijven. 141 De ontwikkeling van de neerwaartse intergenerationele mobiliteit is afge-
beeld in figuur 6. Figuur 6 laat zien dat de neerwaartse intergenerationele mobi-
liteit het spiegelbeeld was van de opwaartse mobiliteit. Analfabete zoons hadden
meer kans om te dalen op de beroepsladder dan alfabete. Toch zijn de verschillen
in mobiliteit tussen alfabeten en analfabeten niet spectaculair groot te noemen. Het valt te verwachten dat ook andere effecten een rol zullen hebben gespeeld. Daarom is ook in dit geval een multivariate analyse uitgevoerd, waarin als onaf-
hankelijke variabelen zijn meegenomen: 1) het (an)alfabetisme van de respon-
dent, 2) het (an)alfabetisme van de vader, 3) de beroepsklasse van de vader, en 4)
de periode waarin de respondent is geboren. Als basis is opnieuw een model gekozen waarin alle hoofd- en eerste-orde
interactie-effecten zijn opgenomen. Met behulp van een logistische-regressieana-
lyse zijn de effecten geschat. Vervolgens zijn via een proces van ‘stepwise back-
ward elimination’ alle niet-significante interactie-effecten uit het model verwij-
derd. Deze procedure is gebruikt voor twee afzonderlijke analyses: een analyse
van opwaartse en een analyse van neerwaartse intergenerationele mobiliteit. In
beide gevallen is de afhankelijke mobiliteitsvariabele als een dichotome varia-
bele gedefinieerd: in het geval van de opwaartse mobiliteit zijn allen die inter- Figuur 6 Intergenerationale neerwaartse beroepsmobiliteit naar alfabetisme van vader en
zoon en geboortejaar (1800-1900)
intergenerationele neerwaartse beroepsmobiliteit van alfabeten en analfabeten Bron: HSN-datasets Akten en Levenslopen, release 2007.01; N = 9.457. Analfabetisme en intergenerationele sociale mobiliteit g
p
0
10
20
30
40
50
60
1800-1825
1825-1850
1850-1875
1875-1900
geboortejaar
% mobiel
beiden analfabeet
vader analfabeet, zoon alfabeet
vader alfabeet, zoon analfabeet
beiden alfabeet beiden analfabeet
vader analfabeet, zoon alfabeet
vader alfabeet, zoon analfabeet
beiden alfabeet beiden analfabeet
vader analfabeet, zoon alfabeet
vader alfabeet, zoon analfabeet
beiden alfabeet Bron: HSN-datasets Akten en Levenslopen, release 2007.01; N = 9.457. functioneel analfabetisme in nederland, 1775-1900 functioneel analfabetisme in nederland, 1775-1900 Tabel 3 Logistische-regressieanalyse van opwaartse en neerwaartse intergenerationele mobilite 3
g
g
y
p
g
Opwaarts mobiel
Neerwaarts mobiel
B
Exp(B)
B
Exp(B)
Intercept
-0,04
-
-0,83**
-
Analfabetisme vader
Analfabeet
-0,72**
0,49
0,37*
1,44
Alfabeet
0
-
0
-
Analfabetisme zoon
Analfabeet
-0,61
0,54
0,49**
1,62
Alfabeet
0
-
0
-
Geboortecohort zoon
1800-1825
0,12
1,13
0,69**
1,98
1825-1850
-0,50
0,95
0,32*
1,37
1850-1875
-0,31
0,73
0,26*
1,29
1875-1900
0
-
0
-
Beroepsklasse vader
Klasse 1
2,04**
7,70
Klasse 2
-2,58**
0,08
1,10**
2,99
Klasse 3
-1,11**
0,33
0,24*
1,28
Klasse 4
-1,18**
0,32
-0,04
0,96
Klasse 5
0,30**
1,48
0
-
Klasse 6
0
-
Geboortecohort * beroepsklasse vader
1800-1825 * Klasse 1
-1,70**
0,18
1825-1850 * Klasse 1
-1,35**
0,26
1850-1875 * Klasse 1
-1,57**
0,21
1875-1900 * Klasse 1
0
-
1800-1825 * Klasse 2
0,64
1,89
-0,58
0,56
1825-1850 * Klasse 2
-0,43
0,65
-0,98**
0,38
1850-1875 * Klasse 2
0,63
1,83
-0,39
0,68
1875-1900 * Klasse 2
0
-
0
-
1800-1825 * Klasse 3
-1,41*
0,24
-0,76*
0,47
1825-1850 * Klasse 3
-0,82*
0,44
-0,51*
0,60
1850-1875 * Klasse 3
-0,23
0,79
-0,62**
0,54
1875-1900 * Klasse 3
0
-
0
-
1800-1825 * Klasse 4
-0,88
0,41
-0,28
0,76
1825-1850 * Klasse 4
-0,60
0,55
-0,29
0,75
1850-1875 * Klasse 4
-0,29
0,75
-0,48**
0,62 142 onno boonstra onno boonstra 143
Opwaarts mobiel
Neerwaarts mobiel
B
Exp(B)
B
Exp(B)
1875-1900 * Klasse 4
0
-
0
-
1800-1825 * Klasse 5
0,33
1,39
0
-
1825-1850 * Klasse 5
0,32
1,38
0
-
1850-1875 * Klasse 5
0,49*
1,64
0
-
1875-1900 * Klasse 5
0
-
0
-
1800-1825 * Klasse 6
0
-
1825-1850 * Klasse 6
0
-
1850-1875 * Klasse 6
0
-
1875-1900 * Klasse 6
0
-
Beroepsklasse vader* analfabetisme zoon
Klasse 1 * analfabeet
Klasse 2 * analfabeet
2,33
10,26
Klasse 3 * analfabeet
1,75
5,73
Klasse 4 * analfabeet
0,63
1,87
Klasse 5 * analfabeet
-0,34
0,71
Klasse 6 * analfabeet
0
-
Klasse 1 * alfabeet
0
-
Klasse 2 * alfabeet
0
-
Klasse 3 * alfabeet
0
-
Klasse 4 * alfabeet
0
-
Klasse 5 * alfabeet
0
-
Klasse 6 * alfabeet
0
-
Geboortecohort * analfabetisme vader
1800-1825 * analfabeet
0,47
1,61
-1,10**
3,00
1825-1850 * analfabeet
0,64*
1,89
-0,27
1,31
1850-1875 * analfabeet
0,12
-1,13
-0,28
1,32
1875-1900 * analfabeet
0
-
0
-
1800-1825 * alfabeet
0
-
0
-
1825-1850 * alfabeet
0
-
0
-
1850-1875 * alfabeet
0
-
0
-
1875-1900 * alfabeet
0
-
0
-
N=7.174
N=5.977
* = p < 0,05; ** = p < 0,01. Bron: HSN-datasets Akten en Levenslopen, release 2007.01. * = p < 0,05; ** = p < 0,01. Bron: HSN-datasets Akten en Levenslopen, release 2007.01. * = p < 0,05; ** = p < 0,01. functioneel analfabetisme in nederland, 1775-1900 Bron: HSN-datasets Akten en Levenslopen, release 2007.01. p < 0,05;
= p < 0,01. functioneel analfabetisme in nederland, 1775-1900 functioneel analfabetisme in nederland, 1775-1900 generationeel opwaarts mobiel waren vergeleken met degenen die niet mobiel
waren; in het geval van de neerwaartse mobiliteit is de procedure omgekeerd. In tabel 3 staan de uitkomsten van de analyse van intergenerationele op- en
neerwaartse mobiliteit vermeld. Bij de opwaartse mobiliteit blijken zowel enige
hoofd- als ook enige interactie-effecten bij te dragen aan de verklaring ervan. Analfabetisme speelt een belangrijke rol als hoofdeffect: wanneer de vader niet
kon lezen en schrijven, maakte dat de kans voor de zoon om te stijgen op de
maatschappelijke ladder een stuk kleiner. Of de zoon zelf ook niet kon lezen en
schrijven, maakte daarbij niet veel uit. Tevens verschilden de kansen op opwaart-
se mobiliteit tussen zonen met vaders uit hogere of lagere klassen: wie uit een
sociaal hoger nest kwam had een minder grote kans om nog verder te stijgen. Gedurende de negentiende eeuw veranderde die situatie niet, hoewel er voor
drie groepen significante interactie-effecten te zien zijn: er is bijvoorbeeld in de
periode 1800-1850 een opvallend negatief effect voor zonen van wie de vader uit
klasse 3 afkomstig was: hun kansen op opwaartse mobiliteit waren significant
lager, terwijl dat effect voor zonen van vaders uit klasse 5 in de periode 1850-
1875 juist groter was. 144 De analyse van neerwaartse intergenerationele mobiliteit levert vier signifi-
cante hoofd-, en twee significante interactie-effecten op. Tabel 3 laat ook zien dat
de kans op neerwaartse mobiliteit toenam wanneer de vader van de respondent
in een van de hoogste klassen vertoefde. Dat is op zichzelf niet onlogisch: de
kans om te dalen is vanzelfsprekend groter naarmate de vader zich in een hogere
klasse bevindt. Naast het hoofdeffect ‘beroep van de vader’ speelde ook zijn ver-
mogen om te kunnen lezen en schrijven een rol. Het analfabetisme van de vader
blijkt namelijk niet alleen de kansen op opwaartse mobiliteit te hebben ver-
kleind, maar ook de kansen op neerwaartse mobiliteit te hebben vergroot. Maar
bij neerwaartse mobiliteit speelde ook het analfabetisme van de zoon een rol: wie
als zoon van een analfabete vader zelf ook niet kon lezen en schrijven, had een
extra grote kans om op de beroepsladder te dalen. Daarnaast maakt tabel 3 dui-
delijk dat de kans op neerwaartse mobiliteit in de loop van de negentiende eeuw
afnam. functioneel analfabetisme in nederland, 1775-1900 Uit het significante interactie-effect van ‘geboortecohort*analfabetisme
vader’ blijkt dat die forse kans op neerwaartse mobiliteit van zonen van anal-
fabete vaders in het eerste kwart van de negentiende eeuw werd gedempt: ken-
nelijk begon het negatieve effect van vaders analfabetisme pas in de loop van de
eeuw een rol van betekenis te spelen. 144 Conclusies De negentiende eeuw is voor ons land een periode geweest van grote economi-
sche, sociale en culturele veranderingen. De economische veranderingen gingen
gepaard met een veranderende sociale stratificatie: sommige beroepsklassen
groeiden fors, andere werden een stuk kleiner. Een belangrijke culturele veran-
dering betekende het verdwijnen van het analfabetisme uit onze samenleving. onno boonstra Degenen die in de laatste helft van de negentiende eeuw werden geboren, zou-
den vrijwel zonder uitzondering de school gaan bezoeken. Dat is in zekere zin
opvallend, omdat er in die tijd nog geen leerplicht bestond. Kennelijk ontwikkel-
de zich bij ouders het idee dat het gewenst was dat hun kinderen leerden lezen
en schrijven om op die manier de modernisering van de samenleving het hoofd
te bieden. In dit hoofdstuk is de vraag gesteld of dit idee in de praktijk zijn weer-
slag heeft gekregen. Het antwoord is tweeledig. Voor een fraaie beroepsloopbaan
maakte het niets uit of iemand kon lezen en schrijven: de kansen om gedurende
de beroepsloopbaan in beroepsklasse te stijgen of te dalen werd niet door het
eigen (an)alfabetisme beïnvloed. Voor een verbetering of verslechtering van de
beroepsklasse ten opzichte van die van de vader, speelde analfabetisme daaren-
tegen wel een rol van betekenis. De kansen om gedurende de beroepsloopbaan
een of meer klassen uit te stijgen boven de beroepsklasse van de vader, waren
kleiner wanneer de vader niet kon lezen en schrijven, terwijl de kansen om te
dalen juist groter waren. De eventuele eigen onmacht om te schrijven maakte de
kans om te dalen nog eens extra groot. Wel kunnen lezen en schrijven zorgde er
daarentegen voor dat de kans op neerwaartse mobiliteit afnam. 145 Op macroniveau heeft het onderwijs zeker bijgedragen aan de sociale, cultu-
rele en economische ontwikkeling van ons land; op microniveau heeft onderwijs
zeer zeker bijgedragen aan de sociale en culturele emancipatie van de individu-
ele burger. Door te kunnen lezen kreeg hij deel aan de culturele opbrengsten van
de samenleving. Het verschafte een zekere waardigheid, net zoals het niet kun-
nen lezen en schrijven langzamerhand iets begon te worden waarvoor men zich
zou moeten schamen. Door naar school te gaan kon het kind ontsnappen aan de
miserabele positie thuis, op het veld of in de fabriek. Het kreeg een glimp van
een perspectief voorgeschoteld waaraan het zich kon vastgrijpen (Vincent, 2000;
Boonstra, 1993). Noten 1. Dr. W. Rutten was zo vriendelijk mij enige pagina’s van dit ongepubliceerde manuscript te
doen toekomen. 1. Dr. W. Rutten was zo vriendelijk mij enige pagina’s van dit ongepubliceerde manuscript te
doen toekomen. 2. Geciteerd in Furet & Ozouf (1977), p. 198. 146 3. Er is één uitzondering waarbij de indeling van HISCLASS niet is gevolgd. Het betreft de
schippers. In HISCLASS zijn die ondergebracht in HISCLASS klasse 4, maar ze horen eerder
thuis in HISCLASS klasse 9. De reden is dat schippers in de negentiende eeuw weliswaar
zelfstandige ondernemers zijn, maar meer als ‘vrachtrijders’ moeten worden beschouwd
dan als kooplieden. 4. In de analyse van de intergenerationele beroepsmobiliteit is de hoogste beroepsklasse van
de zoon vergeleken met de hoogste beroepsklasse van de vader. 4. In de analyse van de intergenerationele beroepsmobiliteit is de hoogste beroepsklasse van
de zoon vergeleken met de hoogste beroepsklasse van de vader. Conclusies Maar dit onderzoek leert dat de verheffing door het onderwijs misschien wel
in culturele zin gestalte heeft gekregen, maar niet in economische. In de samen-
leving konden niet de kansen gerealiseerd worden die het onderwijs veronder-
stelde te kunnen creëren. Wie uit een ‘analfabeet’ nest kwam, begon al met een
achterstand op de arbeidsmarkt, een achterstand die door eigen alfabetisme niet
kon worden ingelopen. Kinderen die hadden leren lezen en schrijven konden
weliswaar iets hoger op de beroepsladder beginnen dan hun (analfabete) vaders
en werden behoed voor een duikeling op de beroepsladder, maar ze kregen geen
duwtje in de rug om een of meer treden te klimmen. Er vallen twee conclusies te trekken. De eerste conclusie is dat ouders de
‘weldaad der scholen’ weliswaar veronderstelden, maar dat hun kinderen die op
de arbeidsmarkt nauwelijks hebben ervaren. En de tweede conclusie die getrok-
ken kan worden is dat het kennelijk economisch gezien even functioneel was
om alfabeet of analfabeet te zijn, omdat de beroepsloopbaan er niet door werd
beïnvloed. Dat betekent dat laaggeletterdheid geen obstakel was om te kunnen
functioneren in de samenleving van de negentiende eeuw. Er bestond – in die
zin althans – functioneel analfabetisme. functioneel analfabetisme in nederland, 1775-1900 Literatuur Arkenbout, P. (z.j.). Korte levensschets van Pieter Arkenbout, door hem uit vroegere
bescheiden in dit boek bijeengebracht in zijn laatste levenstijd, zijnde toen in de Johan-
nes-Stichting te Nieuwveen. Z.p. Arkenbout, P. (z.j.). Korte levensschets van Pieter Arkenbout, door hem uit vroegere
bescheiden in dit boek bijeengebracht in zijn laatste levenstijd, zijnde toen in de Johan-
nes-Stichting te Nieuwveen. Z.p. Arkenbout, P. (z.j.). Korte levensschets van Pieter Arkenbout, door hem uit vroegere
bescheiden in dit boek bijeengebracht in zijn laatste levenstijd, zijnde toen in de Johan-
nes-Stichting te Nieuwveen. Z.p. Barro, R.J. (2001). Human capital and growth. The American Economic Review, 91(2),
12-17. Blau, P.M. & O.D. Duncan (1967). The American occupational structure. New York:
John Wiley & Sons. Boonstra, O.W.A. (1993). De waardij van eene vroege opleiding. Een onderzoek naar de
implicaties van het alfabetisme op het leven van inwoners van eindhoven en omliggende
gemeenten, 1800-1920. Hilversum: Verloren. Boonstra, O.W.A. (2009). Regionale verschillen in de daling van het analfabetisme
in Nederland, 1775-1900. In O.W.A. Boonstra & A.J. Schuurman (red.), Tijd en
ruimte. De toepassing van GIS in de alfa-wetenschappen. Utrecht: Matrijs. Cipolla, C.M. (1969). Literacy and development in the West. Hammondsworth: Pen-
guin Books. Easterlin, R.A. (1981). Why isn’t the whole world developed? The Journal of Econo-
mic History, 41(1), 1-19. Furet, F. & M. Ozouf (1977). Lire et écrire. L’Alphabétisation des Francais de Calvin à Jules
Ferry. Paris: Éditions de Minuit. Graff, H.J. (1979). The literacy myth: literacy and social structure in the nineteenth cen-
tury. New York: Academic Press. Graff, H.J. (red.) (1981). Literacy and social development in the West – a reader. New
York: Cambridge University Press. Graff, H.J. (1987). The legacies of literacy. Continuities and contradictions in Western cul-
ture and society. Bloomington: Indiana University Press. Gylafson, T. (2001). Natural resources, education, and economic development. European Economic Review, 45(406), 847-859. Inkeles, A. (1973). The school as a context for modernization. International Journal
of Comparative Sociology, 14, 163-178. Knippenberg, H. (1986). Deelname aan het lager onderwijs in Nederland gedurende de
negentiende eeuw. Amsterdam: proefschrift. onno boonstra Leeuwen, M.H.D. van, I. Maas & A. Miles (2002). HISCO. Historical International
Standard Classification of Occupations. Leuven: Leuven University Press. Lezen & Schrijven, stichting (2004). Economisch potentieel laaggeletterden onvoldoende
benut in kenniseconomie. 31 december, 2007 ontleend aan http://www.lezenen-
schrijven.nl/files/persberichten/8september2004.pdf. 147 Maas, I. & M.H.D. van Leeuwen (2004, 25 May 2004). HISCO HISCLASS recode scheme. Retrieved 31 december, 2007, from http://historyofwork.iisg.nl/. Literatuur Mandemakers, K. (2001). De sociale structuur in Nederland rond 1900. De samen-
leving in het perspectief van de modernisering 1850-1990. In J.G.S.J. van Maar-
seveen & P.K. Doorn (red.), Nederland een eeuw geteld. Een terugblik op de samenle-
ving rond 1900 (pp. 185-207). Amsterdam: Stichting Beheer IISG. Sanderson, M. (1972). Literacy and social mobility in the industrial revolution in
England. Past & Present, 56, 75-103. Vincent, D. (2000). The rise of mass literacy: reading and writing in modern Europe. Cambridge: Polity Press. Weiner, M. (red.) (1966). Modernization: the dynamics of growth. New York: Basic
Books. Weiner, M. (red.) (1966). Modernization: the dynamics of growth. New York: Basic
Books. Wenken over de opvoeding. (1852). De Economist, pp. 285-290. Wolf, A. (2002). Does education matter? Myths about education and economic growth. London: Penguin. De rol van het gymnasiaal en middelbaar
onderwijs bij de intergenerationele overdracht
van status, Nederland 1865-19401 Richard L. Zijdeman en Kees Mandemakers Inleiding Sinds de jaren negentig van de twintigste eeuw verschaft grootschalige digitali-
sering van historische persoonsgegevens nieuwe mogelijkheden voor onderzoek
naar intergenerationele mobiliteit. Ten eerste geven de historische data een uni-
verseler beeld van het statusverwervingsproces in het verleden. Waar historisch
sociale studies voorheen betrekking hadden op enkele steden over een beperkte
periode, beslaan dergelijke studies nu hele provincies over een periode van meer
dan honderd jaar (Bras & Kok, 2005; Van Leeuwen & Maas, 1995; Zijdeman, 2008). Ten tweede bieden de ‘nieuwe’ data de mogelijkheid theorieën over trendma-
tige ontwikkelingen in de intergenerationele mobiliteit te toetsen. Verschillen
over de tijd in intergenerationele mobiliteit zijn betrekkelijk gering en met data
over langere tijdsperioden zijn deze makkelijker te onderkennen en aan te tonen
(Ganzeboom & Luijkx, 1995; Ganzeboom, Treiman & Ultee, 1991). Zowel periodieke als regionale verschillen in intergenerationele mobiliteit
worden vaak verklaard uit verschillen in de mate van industrialisering en de
daarmee gepaard gaande onderwijsexpansie (Kaelble, 1983; Rijken, 1999; Ringer,
1979). Diverse studies naar de invloed van industrialisering en onderwijs op inter-
generationele mobiliteit maken daarbij gebruik van twee theorieën: de moder-
niseringstheorie en de statusbehoud-theorie. Volgens de moderniseringstheorie
neemt het belang van het ouderlijk milieu voor de statusverwerving af naarmate
de industrialisering voortschrijdt (Blau & Duncan, 1963; Kerr e.a., 1960; Treiman,
1970). Volgens de statusbehoud-theorie blijft dit belang gelijk, al kan de wijze
waarop de invloed plaatsvindt, verschuiven van een directe naar een indirecte
invloed door middel van het onderwijs dat de ouders de kinderen laten volgen
(Collins, 1971, 1979; Grusky 1983). Een directe toets van beide theorieën op basis van data uit historische per-
soonsregisters en censusgegevens is maar beperkt mogelijk. Dergelijke bronnen,
zoals geboorte- en huwelijksakten, bevatten immers geen gegevens over het genoten onderwijsniveau. Wel wordt de aanwezigheid van een handtekening op
een document als indicator voor alfabetisme gebruikt, wat op ten minste enke-
le jaren genoten opleiding zou duiden (Boonstra, 1993, 1995). Een moeilijkheid
van deze indicator is dat het percentage analfabeten reeds halverwege de negen-
tiende eeuw erg gering was en in hoog tempo afnam. In de provincie Utrecht
werd slechts door minder dan 5 procent van de mannen geboren in 1850 geen
handtekening gezet op het huwelijkscertificaat. Dit terwijl er in Utrecht in de
negentiende eeuw toch nog relatief veel analfabetisme was: alleen in Limburg
en Noord-Brabant lagen de percentages structureel hoger (Boonstra, 1993, 1995). Inleiding 150 g
p
g
g
(
)
Wanneer data uit historische persoonsregisters worden gecombineerd met
archiefdata over schoolloopbanen is een directe toets van de moderniserings-
en de statusbehoud-theorie wel mogelijk. Van Dijk en Mandemakers (1985)
doen dit voor twee cohorten leerlingen (1880-’81 en 1920) uit het gymnasiaal en
middelbaar onderwijs te Rotterdam (in de terminologie van voor de invoering
van de Mammoetwet in 1968: het voorbereidend hoger en middelbaar onder-
wijs (vhmo)). Zij verzamelden data over leerlingen op het individuele niveau uit
schoolarchieven (zoals leeftijd, adres, naam ouders) en combineerden deze ver-
volgens met informatie uit de bevolkingsregisters. Van veel belang zijn verder de
studie van Harrigan (1980) naar de leerlingen van het Franse middelbare onder-
wijs, op basis van een bewaard gebleven enquête uit 1864 van de toenmalige
minister van Onderwijs Victor Duruy, en de studie van Lundgreen, Kraul en Ditt
(1988) naar de onderwijskansen en sociale mobiliteit van kinderen in Duisburg
en Minden tussen 1860 en 1918. Deze studie volgt een soortgelijke aanpak en behandelt daarbij de volgende
vragen: g
In welke mate veranderde de associatie tussen beroepsstatus van vaders en
1. zonen tussen de beide cohorten 1880-’81 en 1920 in Nederland? In hoeverre zijn veranderingen in deze associatie te verklaren door expansie
2. van het voorbereidend hoger en middelbaar onderwijs (vhmo)? Door antwoord te geven op bovenstaande vragen draagt deze studie op twee
manieren bij aan het bestaande onderzoek naar intergenerationele statusver-
werving. In deze studie wordt namelijk voor het eerst een vergelijking gemaakt
tussen de stratificatieprocessen van mensen met en zonder een vhmo-opleiding
aan het eind van de negentiende en het begin van de twintigste eeuw. Eerdere
historische studies bevatten veelal geen informatie over genoten opleiding of
beperkten zich juist uitsluitend tot vhmo-leerlingen (Van Dijk & Mandemakers,
1985). Daarnaast beperkt eerder onderzoek zich veelal tot enkele steden. Dit
onderzoek daarentegen maakt gebruik van datasets die grote delen van Neder-
land omvatten, waardoor de resultaten generaliseerbaar zijn naar het nationale
niveau. Het gaat hier om een representatieve steekproef van vhmo-leerlingen uit
de cohorten 1880-’81 en 1920 (Mandemakers, 1996a) en een vergelijkbaar bestand
uit de Historische Steekproef Nederlandse bevolking (HSN). Omdat in het data- richard l. zijdeman en kees mandemakers bestand van vhmo-leerlingen uitsluitend jongens zijn opgenomen, moet ook de
vergelijkende HSN-studie zich tot het mannelijke geslacht beperken. De cohor-
ten in beide bestanden starten op het moment dat de leerlingen in de eerste klas
begonnen. Inleiding Dit was gemiddeld op dertienjarige leeftijd. De huwelijksleeftijd lag
gemiddeld tussen de 25 en 35 jaar. Dit betekent dat deze studie grosso modo de
periode 1865-1940 beslaat. 151 Het statusverwervingsproces: twee interpretaties Het statusverwervingsproces: twee interpretaties
In hun klassieker The American Occupational Structure introduceerden Blau en
Duncan (1967) het statusverwervingsmodel. In de meest eenvoudige vorm van
het model wordt beroepsstatus van een zoon direct en indirect beïnvloed door
de beroepsstatus van zijn vader (figuur 1). Pijl A geeft het directe verband weer. Hierbij kan men denken aan de invloed van de sociale achtergrond op de beroeps-
keuze van de zoon, bijvoorbeeld heel direct aan de mogelijkheid dat de zoon de
vader in zijn eigen bedrijf opvolgt. De indirecte invloed van vaders beroepsstatus
loopt via de opleiding van de zoon. De beroepstatus van de vader heeft invloed
op het opleidingsniveau van de zoon (pijl B), terwijl het opleidingsniveau weer
invloed uitoefent op de beroepsstatus van de zoon (pijl C). Figuur 1 Eenvoudig statusverwervingsmodel. Pijlen geven positieve causale relaties weer Figuur 1 Eenvoudig statusverwervingsmodel. Pijlen geven positieve causale relaties weer Figuur 1 Eenvoudig statusverwervingsmodel. Pijlen geven positieve causale relaties weer
In de literatuur bestaan sinds geruime tijd twee theorieën die een gelijk beeld
geven van het statusverwervingsproces in pre-industriële samenlevingen (figuur
2a), maar tegenstrijdige verwachtingen hebben over de invloed van industrialise-
ring en andere moderniseringsprocessen op het statusverwervingsproces (figuur
2b en 2c). De moderniseringstheorie (Blau & Duncan, 1967; Kerr e.a., 1960; Trei-
man, 1970) en de statusbehoud-theorie (Bourdieu & Passeron, 1977; Collins,
1971; Grusky, 1983) verwachten beide dat er in pre-industriële samenlevingen
een sterk positief verband is tussen zowel de beroepsstatus van de vader als de
Beroepsstatus
vader
Beroepsstatus
zoon
Opleiding
zoon
A
B
C Figuur 1 Eenvoudig statusverwervingsmodel. Pijlen geven positieve causale relaties weer
Beroepsstatus
vader
Beroepsstatus
zoon
Opleiding
zoon
A
B
C Beroepsstatus
vader Beroepsstatus
zoon Opleiding
zoon Opleiding In de literatuur bestaan sinds geruime tijd twee theorieën die een gelijk beeld
geven van het statusverwervingsproces in pre-industriële samenlevingen (figuur
2a), maar tegenstrijdige verwachtingen hebben over de invloed van industrialise-
ring en andere moderniseringsprocessen op het statusverwervingsproces (figuur
2b en 2c). De moderniseringstheorie (Blau & Duncan, 1967; Kerr e.a., 1960; Trei-
man, 1970) en de statusbehoud-theorie (Bourdieu & Passeron, 1977; Collins,
1971; Grusky, 1983) verwachten beide dat er in pre-industriële samenlevingen
een sterk positief verband is tussen zowel de beroepsstatus van de vader als de de rol van het gymnasiaal en middelbaar onderwijs beroepsstatus van de zoon, en zo ook tussen de beroepsstatus van de vader en de
opleiding van de zoon (pijl A en B in figuur 2a). Het statusverwervingsproces: twee interpretaties Niet alleen volgden zonen hun
vaders dikwijls op in hun beroep, ook werden zij ‘geschoold’ door van kinds af
aan te helpen bij het werk van hun ouders. Vanwege de sterke overeenkomst tus-
sen het door het ouderlijke huis bepaalde latere beroep en de daarop voorberei-
dende opleiding, wordt door beide theorieën een minder sterk effect verwacht
van opleiding (pijl C in figuur 2a) in de periode voor de industrialisatie. 152 p
g (p j
g
)
p
De verschillende visies van de moderniseringstheorie en statusbehoud-theo-
rie op geïndustrialiseerde samenlevingen worden weergegeven in respectievelijk
figuur 2b en 2c. Volgens de moderniseringstheorie neemt tijdens en na de indus-
trialisatie het directe verband tussen de beroepsstatus van vaders en zonen af. De mechanisering van arbeid doet traditionele beroepen verdwijnen, waardoor
beroepen steeds minder van vader op zoon kunnen worden overgedragen. Daar-
naast vindt er een verplaatsing van de productie van goederen naar de productie
van diensten plaats (Kuznets, 1957; Van Zanden & Griffiths, 1989). Met de ont-
wikkeling van een complex productie-, distributie- en marketingsysteem nam
de vraag naar technisch en administratief personeel toe (Treiman, 1970). Zonen
kunnen hun vaders dus steeds minder opvolgen in traditionele beroepen, terwijl
er wel ruimte voor hen is in ‘nieuwe’ beroepen. Voor deze nieuwe beroepen is
echter kennis vereist die niet binnen de familie kan worden overgedragen. Daar-
door vermindert ook de invloed van achtergrond op de opleiding van de zoon. Deze processen vonden ook in Nederland plaats. Vanuit een in 1850 al relatief
sterke dienstenstructuur nam het aandeel van op kennis gebaseerde beroepen
vooral na 1900 sterk toe (Mandemakers, 2001; Van Zanden & Van Riel, 2000). De
economische ontwikkeling bracht een enorme groei van het onderwijs met zich
mee, waardoor alleen al de sociale achtergrond van de kinderen minder elitaire
vormen aannam (Mandemakers, 1999; Van der Ploeg, 1993). Aan de hand van de
moderniseringstheorie leiden we de volgende hypothesen af: H1 De directe invloed van vaders beroepsstatus op die van de zoon is in 1920 geringer dan
in 1880 H1 De directe invloed van vaders beroepsstatus op die van de zoon is in 1920 geringer dan
in 1880 H2 De invloed van vaders beroepsstatus op de opleiding van de zoon is in 1920 geringer
dan in 1880 De moderniseringstheorie veronderstelt verder dat er een verschuiving in waar-
den plaats had. Het statusverwervingsproces: twee interpretaties Waar voor de industrialisering met name afkomst van belang
was, zou met de industrialisering de waardering voor verworvenheden groter
worden (Parsons & Shils, 1951). Op de arbeidsmarkt zou daardoor selectie steeds
meer plaats hebben op basis van eigen prestaties en steeds minder op basis van
afkomst. Hierdoor zou het belang van onderwijs toenemen (vergelijk de dubbele
pijl in figuur 2b). richard l. zijdeman en kees mandemakers richard l. zijdeman en kees mandemakers H3 De invloed van zoons opleiding op zoons beroepsstatus is in 1920 groter dan in 1880 Figuur 2 Twee interpretaties van het statusverwervingsmodela
a De figuur geeft de relaties binnen het statusverwervingsmodel weer in een pre-industriële samen-
leving (figuur 2a), in een industriële samenleving volgens de moderniseringstheorie (figuur 2b) en in
een industriële samenleving volgens de statusbehoud-theorie (figuur 2c). Alle pijlen geven positieve
causale relaties weer. Meerdere pijlen geven sterkere relaties weer. moderniseringstheorie
statusbehoud-theorie
vader
Opleiding
zoon
Industriële samenleving
Pre-industriële samenleving
Fig. 2a
Beroepsstatus
zoon
Beroepsstatus
vader
Opleiding
zoon
Beroepsstatus
zoon
Beroepsstatus
Beroepsstatus
vader
Beroepsstatus
zoon
Opleiding zoon
Fig. 2b
Fig. 2c 1
Figuur 2 Twee interpretaties van het statusverwervingsmodela
Pre-industriële samenleving
Fig. 2a
Beroepsstatus
vader
Beroepsstatus
zoon
Opleiding zoon Figuur 2 Twee interpretaties van het statusverwervingsmodela 153 Pre-industriële samenleving Beroepsstatus
zoon Beroepsstatus Beroepsstatus
vader vader Opleiding zoon Fig. 2a Fig. 2a Industriële samenleving moderniseringstheorie
statusbehoud-theorie
vader
Opleiding
zoon
g
Beroepsstatus
zoon
Beroepsstatus
vader
Opleiding
zoon
Beroepsstatus
zoon
Beroepsstatus
Fig. 2b
Fig. 2c statusbehoud-theorie moderniseringstheorie zoon
Beroepsstatus vader
Beroepsstatus Opleiding Opleiding
zoon Opleiding Fig. 2c a De figuur geeft de relaties binnen het statusverwervingsmodel weer in een pre-industriële samen-
leving (figuur 2a), in een industriële samenleving volgens de moderniseringstheorie (figuur 2b) en in
een industriële samenleving volgens de statusbehoud-theorie (figuur 2c). Alle pijlen geven positieve
causale relaties weer. Meerdere pijlen geven sterkere relaties weer. Een andere visie op de rol van onderwijs tijdens de industrialisatie geeft de sta-
tusbehoud-theorie (Grusky, 1983). Deze heeft geen expliciete verwachtingen over
een afname van het directe verband tussen beroepsstatus van vader en zoon. De
theorie stelt echter wel dat in samenlevingen met afnemende rechtstreekse over-
drachten, de ouders zullen gaan investeren in langduriger en kwalitatief hoger
onderwijs voor hun nageslacht. Aangezien elites meer middelen hebben en de
cultuur binnen het onderwijs meer zou aansluiten bij die van de hogere soci-
ale klassen, zouden kinderen met een hogere sociale achtergrond succesvoller
zijn in het onderwijstraject (Bourdieu, 1977). Ook kunnen zij meer investeren in de rol van het gymnasiaal en middelbaar onderwijs de omstandigheden waaronder hun kinderen studeren, zoals dagelijkse onkos-
ten en woonruimte (Dronkers & De Graaf, 1995). Hierdoor zal de invloed van de
beroepsstatus van de vader op de opleiding van de zoon toenemen. Context en eerdere bevindingen Het vhmo in Nederland eind negentiende en begin twintigste eeuw Het vhmo in Nederland eind negentiende en begin twintigste eeuw
Omstreeks 1850 kon men in Nederland voor middelbaar onderwijs vooral terecht
op de zogenaamde Franse scholen. Deze vormden een naar kwaliteit van het
onderwijs en omvang van de scholen sterk variërend geheel. Daarnaast waren
er de Latijnse scholen die voorbereidden op de universiteit en waarvan een klein
aantal in de grote steden tussen 1840 en 1850 ook tweede afdelingen oprichtte. Op deze afdelingen lag de nadruk op moderne talen en de exacte vakken. Samen
met de betere gemeentelijke Franse scholen vormden deze tweede afdelingen de
basis voor de expansie van de hogere burgerschool (hbs). De hbs liet al snel na
de aanvaarding in 1863 van de Wet op het Middelbaar Onderwijs een behoorlijk
dekkend, over het gehele land verspreid netwerk, zien. De bestaande Latijnse
scholen konden de concurrentie met de nieuwe hbs moeilijk aan en verdwenen
of werden met de Wet van 1876 op het Hoger Onderwijs hervormd tot gymnasia. Bij deze Latijnse scholen nieuwe stijl werd in grote mate het programma van
de hbs als voorbeeld genomen. Beide institutionele hervormingen vormden een
solide basis voor de ontwikkeling van zowel de hbs als het gymnasium, en voor
verwante schooltypen zoals de hbs met driejarige cursus en de middelbare meis-
jesscholen (msvm). Al deze schooltypen bij elkaar werden tot de invoering van de
Mammoetwet in 1968 aangeduid als voorbereidend hoger en middelbaar onder-
wijs (vhmo), de verzamelterm die wij ook in dit hoofdstuk hanteren. 155 In 1880 bestond het vhmo uit ongeveer honderd scholen waaronder 27
gemeentelijke gymnasia en 37 ook meestal gemeentelijke hbs’en met vijfjarige
cursus en twaalf gemeentelijke middelbare meisjesscholen. Bij elkaar telde het
vhmo ongeveer 8.000 leerlingen. Na 1900 en vooral 1910 breidde het aantal leer-
lingen zich snel uit. Deze groei viel samen met de oprichting van binnen het
kader van het vhmo passende schooltypen zoals handelsdagscholen en handels-
afdelingen die als kopklassen boven de eerste drie jaar van de hbs fungeerden. Na 1920 vond de groei vooral plaats op lycea, een sinds 1907 bestaande combi-
natie van gymnasium en hbs met een gezamenlijke een- of tweejarige onder-
bouw. De groei van het vhmo werd sterk gesteund door de rijksoverheid en hing
samen met de industrialisering zoals die in Nederland plaatsvond tussen 1895
en het begin van de Eerste Wereldoorlog. richard l. zijdeman en kees mandemakers 154 H4 De invloed van vaders beroepsstatus op opleiding van de zoon is in 1920 groter dan in
1880 1880 Verder stelt de statusbehoud-theorie dat het onderwijs voor de elite een middel
is om statusverschillen in stand te houden. Via onderwijs krijgt men bepaalde
waarden mee, leert men te spreken over bepaalde onderwerpen en deelt men
activiteiten met een bepaalde groep (Collins, 1971). De lengte en het type onder-
wijs dat men volgt, heeft daarom consequenties voor de cultuur die men zich
verwerft. Collins stelt dat werkgevers op die manier via onderwijskwalificaties
werknemers kunnen selecteren op basis van bepaalde gewenste culturele ach-
tergronden: ‘Educational requirements for employment can serve both to select
new members for elite positions who share the elite culture and, at a lower level
of education, to hire lower and middle employees who have acquired a general
respect for these elite values’ (Collins, 1971, p. 1011). Hieruit maken we op dat de
moderniseringshypothese onder H3 ook opgaat voor de statusbehoud-theorie. H5 De invloed van zoons opleiding op zoons beroepsstatus is in 1920 groter dan in 1880 Figuur 2c laat zien hoe volgens de statusbehoud-theorie de verhoudingen in het
statusverwervingsmodel verschuiven als gevolg van industrialisering en onder-
wijsexpansie. j
p
In de introductie hebben we gesteld dat bij een directe toets van de moder-
niserings- en statusbehoud-theorie de invloed van opleiding als variabele in het
model opgenomen dient te worden. Nu kan duidelijk gemaakt worden waar-
om. Zowel de moderniseringstheorie als de statusbehoud-theorie stellen dat de
samenhang tussen de onderdelen van het statusverwervingsmodel zal verande-
ren. Of de totale associatie tussen beroepsstatus van vader en zoon daarmee ook
verandert, hangt af van de grootte van deze veranderingen. Een toe- of afname
van het totale verband hangt immers af van de verhouding tussen de afnemende
invloed van de beroepsstatus van de vader op het opleidingsniveau van de zoon
en de toenemende invloed van het bereikte onderwijsniveau op de beroepstatus
van de zoon. Over de grootte van de verschillen in de onderdelen van het status-
verwervingsmodel doen beide theorieën echter geen uitspraken. Daarom moet
men, wanneer het bereikte onderwijsniveau niet in de analyses is opgenomen,
veronderstellingen maken over de grootte van de deeleffecten in het statusver-
wervingsmodel (Treiman, 1970). Door onderwijs wel op te nemen in de analyse
kunnen de beide theorieën worden getoetst zonder deze aannames. richard l. zijdeman en kees mandemakers Context en eerdere bevindingen De stimulans van de centrale overheid
bestond vooral uit het oprichten van rijks-hbs’en en het op deze scholen beschik-
baar stellen van kosteloze plaatsen. In 1915 kwam er een algemenere regeling
waarbij het schoolgeld werd gekoppeld aan het ouderlijke inkomen. Vergelijk-
bare regelingen werden al spoedig door het gemeentelijk en – in mindere mate –
ook door het bijzonder onderwijs overgenomen. Omstreeks 1920 waren er 37.000
leerlingen welk aantal steeg tot 210.000 in 1965. Met de invoering van de Mam-
moetwet kwam er een eind aan de hbs en de daarmee verwante schooltypen met
uitzondering van het gymnasium, dat in veel gemeenten als aparte categorale
school bleef bestaan (Mandemakers, 1996a, 1999). de rol van het gymnasiaal en middelbaar onderwijs Dit hoofdstuk heeft betrekking op de leerlingen van het gymnasium en de
hbs. Dit waren overigens niet de enige vormen van secundair onderwijs. Behalve
het beroepsonderwijs bestond het secundair onderwijs ook nog uit een deel dat
sinds de Mammoetwet wordt aangeduid met ‘algemeen vormend’. Voor 1968
werd dit soort onderwijs tot het lager onderwijs gerekend. De Wet op het Lager
Onderwijs van 1857 maakte niet alleen scholen met gewoon lager onderwijs
mogelijk, maar ook scholen met meer uitgebreid lager onderwijs (mulo’s). De
mulo bouwde niet alleen voort op de vakken van het lager onderwijs, maar men
gaf ook les in de moderne talen en wiskunde. De wetgeving dekte een zeer geva-
rieerde praktijk. Rond 1880 waren er maximaal 12.000 leerlingen op dit soort
scholen, een aantal dat pas na 1910 duidelijk ging groeien om in 1920 op 65.000
leerlingen uit te komen. Lang niet alle leerlingen deden echter het mulo-exa-
men, ongeveer de helft bestond uit leerlingen die er een zevende of achtste klas
bij deden, bovenop het curriculum van de gewone lagere school. Bij het beroeps-
onderwijs begon de groei van het aantal scholen pas na 1900. Omstreeks 1920 tel-
den ook de ambachtsscholen en de huishoudscholen ongeveer 20.000 leerlingen
(dagonderwijs). Voor een volledig beeld van het onderwijs aan twaalf- tot en met
achttienjarigen moeten ook nog het middelbaar beroepsonderwijs en de kweek-
scholen met in 1920 ongeveer 10.000 leerlingen meegerekend worden (Boekholt
& De Booij, 1987; CBS, 1959; Mandemakers 1996a). 156 Eerdere bevindingen Op basis van eerdere studies kunnen nog geen algemene conclusies over het sta-
tusverwervingsproces in Nederland voor de Tweede Wereldoorlog worden getrok-
ken. Enerzijds zijn er grote regionale, temporele, methodische en populatiever-
schillen tussen de in aantal beperkte studies die voorhanden zijn (Boonstra &
Mandemakers, 1995; Van Dijk & Mandemakers, 1985; Van Leeuwen & Maas, 1997). Anderzijds betrekken slechts twee studies de invloed van opleiding van de zoon
in hun analyses. De studies die er zijn laten zien dat er in de negentiende eeuw
geen algehele trend naar meer totale mobiliteit was en dat toenames in totale
mobiliteit niet samenhingen met industrialisatie (Boonstra & Mandemakers,
1995; Van Leeuwen & Maas, 1997). Daarnaast laat een studie naar de provincie
Utrecht zien dat er geen duidelijke toename is in de relatieve mobiliteit (Van
Leeuwen & Maas, 1997). Zijdeman (2007) vindt ook voor de provincie Zeeland
geen veranderingen over de tijd in de totale associatie tussen beroepsstatus van
vaders en zonen, noch een verband met urbanisatie, in de late negentiende en
vroege twintigste eeuw. Slechts twee studies bestuderen expliciet de invloed van opleiding van de
zoon in het statusverwervingsproces. In de studie van Van Dijk en Mandemakers
(1985) naar het vhmo en sociale mobiliteit in Rotterdam verschillen de padcoëffi-
ciënten tussen de cohorten 1880-’81 en 1920 niet significant van elkaar. Het gaat
hierbij om het effect van vaders beroepsstatus op die van de zoon en op die van
het schooltype van de zoon. Eveneens is de invloed van het schooltype (hbs 3- of richard l. zijdeman en kees mandemakers 5-jarige cursus of gymnasium) op de beroepsstatus van de zoon niet verschillend
tussen 1880-’81 en 1920. Boonstra (1993) bestudeert het padmodel zoals weer-
gegeven in figuur 1 met als achterliggende verklarende variabele de opleiding
van de vader. De opleiding van zowel de vader als de zoon is gemeten door te kij-
ken of men een handtekening kon zetten of niet. De studie omvat Eindhoven en
enkele omliggende gemeenten aan het eind van de achttiende en gedurende de
hele negentiende eeuw. Helaas zijn in de tabel met uitkomsten geen standaard-
fouten voorhanden, waardoor het lastig is zelf conclusies te trekken over signifi-
cante veranderingen in het statusverwervingsmodel over de tijd (Boonstra, 1993,
tabel 7.14, p. 209). Boonstra rapporteert voor de gehele periode een belangrijke
in de tijd afnemende directe samenhang tussen de sociale status van de vader
en die van de zoon. Eerdere bevindingen Het directe negatieve effect van een analfabete vader neemt
toe gedurende de negentiende eeuw, terwijl het indirecte effect (via de status
van de vader) afneemt. Het directe positieve effect van enige opleiding van de
zoon zelf op zijn latere status neemt af na 1850 (p. 207). Deze afname kan een
inhoudelijke betekenis hebben, maar kan ook een gevolg zijn van het verdwij-
nende analfabetisme. Hierdoor boet alfabetisme als onderscheidend kenmerk
voor het verkrijgen van een relatief hoge beroepsstatus aan belang in. Boonstra
stelt daarom voor opleiding van de zoon aan het einde van de negentiende eeuw
op een andere wijze te meten (p. 210). 157 Data en methoden Data Data Om de invloed van onderwijs te schatten in het eenvoudige statusverwervings-
model is idealiter een nationaal breed onderzoeksbestand nodig met daarin
zowel gegevens over beroepsstatus van vaders en zonen als gegevens over het
door de zonen gevolgde onderwijs. Helaas is een dergelijk databestand niet voor-
handen. Een analyse van het mobiliteitsproces in Nederland aan het eind van de
negentiende en begin van de twintigste eeuw is wel mogelijk door een bestaand
bestand met gegevens van vhmo-leerlingen te vergelijken met een vergelijkbare
steekproef uit de HSN. Om de mobiliteit op een vergelijkbare wijze te meten werd
gebruikgemaakt van huwelijksakten. Voor het vhmo-databestand zijn deze akten
erbij gezocht, voor de HSN waren de akten reeds aanwezig. Hoewel huwelijksak-
ten een unieke bron zijn om intergenerationele mobiliteit te bestuderen en vele
recente studies er gebruik van maken (Van Leeuwen & Maas, 2007; Van Poppel,
Monden & Mandemakers, 2008), is deze toepassing in het verleden bekritiseerd
(Delger & Kok, 1998). Een eerste punt van kritiek richt zich erop dat het beroep
van de vader van de bruidegom op een later moment in zijn carrière gemeten
is dan het beroep van de bruidegom zelf. In onze studie gaat het echter om een
vergelijking tussen cohorten, waardoor het vooral van belang is dat de beroe-
pen tussen de cohorten op hetzelfde tijdstip zijn gemeten. Huwelijksakten en de rol van het gymnasiaal en middelbaar onderwijs dan vooral die van eerste huwelijken waarbij bruidegoms aan het begin van hun
carrière staan, zijn daarvoor uitermate geschikt. Een tweede punt van kritiek
stelt dat het ontbreken van beroepstitels van de vaders op huwelijksakten niet
random is, maar voornamelijk samenhangt met vroegtijdig overlijden. Wanneer
aangenomen wordt dat dit vroegtijdig overlijden negatief samenhangt met soci-
ale achtergrond, dan zou het gebruik van huwelijksakten leiden tot een onder-
vertegenwoordiging van mensen met een lagere sociale status. Deze aanname
lijkt op basis van onze data echter niet gerechtvaardigd. Bij de bespreking van de
datasets hieronder zal blijken dat in de dataset van de vhmo-leerlingen, waarin
de bovenlaag van de samenleving oververtegenwoordigd is, het percentage ont-
brekende beroepstitels niet veel minder is dan bij het HSN-bestand, dat een ruwe
doorsnede van de gehele bevolking is. 158 Het vhmo-databestand werd verzameld voor het onderzoek van Mandema-
kers (1996a) naar de sociale achtergrond en onderwijsloopbanen van leerlingen
van vhmo-scholen. De steekproef beperkte zich tot de op deze scholen aanwezige
jongens. Data Deze beperking werd vooral ingegeven door de oorspronkelijk sterke
gerichtheid van dit onderzoek op de relatie tussen onderwijs en beroepsmobili-
teit. Op basis van eerdere studies was het duidelijk dat dit in de late negentiende
eeuw en vroege twintigste eeuw vrijwel uitsluitend jongens betrof. Sociale mobi-
liteit bij meisjes zou vooral gaan om huwelijksmobiliteit en niet om beroeps-
mobiliteit (Van Dijk & Mandemakers, 1985; Mandemakers, 1996b, 181-182). In
het onderzoek werd gekozen voor een cohortbenadering waarin drie generaties
met elkaar werden vergeleken (1880, 1920 en 1965). Het jaar 1880 werd gekozen,
omdat in dat jaar het vhmo behoorlijk was uitgekristalliseerd. Het cohort 1920
kwam na de sterke expansie van het aantal leerlingen tussen 1900 en 1920. Het
cohort 1965 was het laatste cohort voor de invoering van de Mammoetwet waar
reeds een databestand van bestond. De gegevens voor de cohorten 1880 en 1920 werden uit de archieven van de
desbetreffende scholen gehaald. De beschikbaarheid van archiefmateriaal was
dusdanig, 80 procent van alle scholen uit 1880 en 70 procent van die uit 1920,
dat het mogelijk was om een steekproef te trekken die als een nationale steek-
proef kon gelden. In de steekproef werden die leerlingen opgenomen die op 1
september in 1880 en 1920 voor het eerst in de eerste klas van het vhmo werden
opgenomen. In verband met betere steekproefverhoudingen werd de steekproef
1880 getrokken uit zowel 1880 en 1881 (hierna cohort 1880-’81). In deze studie
wordt gebruikgemaakt van de nationale steekproef (1880-’81: n = 1037; 1920: n =
926; de steekproeffractie bedraagt respectievelijk 23,4 % en 12,5 %) (Mandema-
kers, 1996b, p. 191). Om met het HSN-bestand vergelijkbare informatie te krijgen over het beroep
van deze leerlingen hebben wij zoveel mogelijk getracht de huwelijksplaats
en datum te achterhalen van de vhmo-leerlingen.2 Hierbij is ook gezocht naar
andere levensloopinformatie, bijvoorbeeld om te achterhalen of leerlingen
ongehuwd waren overleden. Deze informatie is via diverse digitale historische richard l. zijdeman en kees mandemakers persoonsarchieven verkregen.3 Daarnaast geven de digitale bronnen informatie
over waar de akte is opgemaakt. Deze gegevens zijn gebruikt om de akten in de
provinciale en gemeentearchieven te lokaliseren. Vervolgens zijn de huwelijks-
akten opgespoord en ingevoerd.4 Van de in totaal 1853 leerlingen zijn voor 261
leerlingen aanwijzingen gevonden dat ze nooit zijn getrouwd. Van de overige
1592 is voor 1156 leerlingen met zekerheid vastgesteld dat ze ten minste een-
maal getrouwd zijn. Data Van deze leerlingen werden 990 huwelijksakten van eerste
huwelijken teruggevonden met een beroepstitel van de bruidegom. Helaas bleek
bij controle dat voor 19 leerlingen niet met zekerheid gesteld kon worden of ze
hadden deelgenomen aan erkend voorbereidend hoger en middelbaar onderwijs. Daarnaast ontbrak in bijna de helft van de gevallen de beroepstitel van de vader. Omdat bij huwelijken niet alle vaders meer in leven zijn of omdat vaders gepen-
sioneerd zijn, was er slechts voor 447 akten een beroepstitel van zowel zonen als
vaders voorhanden. Dit gaat dus tegen de verwachting van Delger en Kok in, dat
onder vaders van hogere sociale afkomst, de uitval van beroepstitels geringer
zou zijn (Delger & Kok, 1998). 159 Om meer cases over te houden gebruiken we daarom een eerdere meting
van vaders beroep, namelijk als de zoon ongeveer dertien jaar oud is en binnen-
treedt in het vhmo. Informatie over dit beroep is afkomstig uit bevolkingsregis-
ters en belastingkohieren (Mandemakers, 1996a, 215-218). Deze alternatieve titels
zijn gemiddeld vijftien jaar eerder gemeten dan de beroepstitels op de huwe-
lijksakten. Op dat moment waren de vaders dus minder vergevorderd in hun
carrière dan bij de meting op de huwelijksakte. Daarom gaan we na in hoeverre
de beroepsstatus van de vervangende beroepstitels van vaders afwijkt van die
van de vaders zoals vermeld op de huwelijksakten. De gemiddelde beroepsstatus
van vaders van wie het beroep op de huwelijksakte was geregistreerd, bedraagt
70,1 op de HIS-CAM schaal met een standaard deviatie van 16,3. De gemiddelde
beroepsstatus van vaders met een alternatieve meting van de beroepsstatus is
69,6 met een standaarddeviatie van 16,4. Deze verschillen lijken marginaal en
een student t-toets wijst dan ook uit dat de verschillen in het gemiddelde niet
significant zijn (t = 0,396; p = 0,693), terwijl een Levenes test dit ook aantoont
voor de verschillen in de variantie (f = 0,982, p = 0,850). We gaan er dan ook van
uit dat het gebruik van deze vervangende beroepstitels de uitkomsten van de
analyse niet significant beïnvloedt. Door het gebruik van de alternatieve meting
van beroepstitels van vaders konden in totaal 878 akten gebruikt worden voor
de analyses. Daarmee komt de ‘respons’ ten opzichte van alle mogelijk gehuwde
vhmo-leerlingen uit op 58,9 procent en ten opzichte van de vhmo-leerlingen met
een gevonden huwelijksakte op 77,2 procent. Data De tweede dataset waar we gebruik van maken is een release van huwelijks-
akten uit de HSN (HSN-dataset Akten burgerlijke stand release 2007.01). Uiteindelijk
zal de dataset alle huwelijken omvatten van de personen uit de HSN-steekproef. De hier gebruikte release bevat huwelijken uit alle provincies, maar huwelijken
uit de provincies Zeeland, Utrecht en Friesland en huwelijken van mensen die in de rol van het gymnasiaal en middelbaar onderwijs dezelfde provincie trouwen als waarin ze geboren zijn, zijn oververtegenwoor-
digd. Om een bestand te krijgen dat vergelijkbaar is met dat van de vhmo-leer-
lingen selecteerden we uit dit bestand de eerste huwelijken van bruidegoms die
tussen 1875 en 1885 en 1915 en 1925 dertien jaar oud waren. Deze leeftijd is
gekozen, omdat dit zowel in de vhmo-cohorten 1880-’81 als in die van 1920 de
modus van de leeftijd van intrede in het vhmo was (Mandemakers, 1996b, tabel
14.6, p. 302). De dataset omvat dan 3219 huwelijksakten met een vermelding van
het beroep van de bruidegom. Ook op deze akten is slechts voor een beperkt
deel, namelijk 1854 gevallen (57,6 %), het beroep van de vader vermeld. 160 j
g
(
)
p
In de analyses worden drie variabelen gebruikt: beroepsstatus van de zoon,
beroepsstatus van de vader en opleiding van de zoon. Beroepsstatus van zoon en vader
is verkregen door alle beroepstitels te coderen in de Historical International
Standard Classification of Occupations (HISCO) (Van Leeuwen, Maas & Miles,
2002). HISCO is een beroepenclassificatie voor historische beroepen gebaseerd
op de International Standard Classification of Occupations 1968 van het Inter-
national Labour Office (ISCO68, 1969). Door middel van deze classificatie wordt
aan beroepen met soortgelijke taken en activiteiten een gelijke code toegekend. Om de beroepen te ordenen naar status gebruiken we een historische continue
beroepsprestigeschaal: HIS-CAM versie 0.1 (Maas e.a., 2006; Lambert e.a., 2006). HIS-CAM is een historische versie van de CAMSIS-schalen, die veronderstellen
dat patronen van sociale interactie tussen mensen uit verschillende beroepscate-
gorieën representatief zijn voor de gehele beroepsstratificatiestructuur. De HIS-
CAM-schaal is geconstrueerd aan de hand van associaties tussen beroepstitels
afkomstig van 1,5 miljoen huwelijksakten uit 6 verschillende landen (Canada,
Duitsland, Frankrijk, Groot-Brittannië, Nederland en Zweden) uit de periode
1800-1938. De associaties zijn geschat met Goodmans RCII-modellen (Goodman,
1979). De schaalscores variëren tussen 0 en 100. Data De variabele opleiding van de
zoon geeft weer of de zonen geen vhmo gevolgd hebben (0), ingeschreven ston-
den bij een driejarige hbs (1), bij een vijfjarige hbs (2) of bij een gymnasium (3). Deze onderverdeling in vhmo-typen is afkomstig uit Mandemakers (1996a). Door Tabel 1 Verdeling van het aantal cases (absoluut en procentueel) over cohorten en datasets el 1 Verdeling van het aantal cases (absoluut en procentueel) over cohorten en datasets Tabel 1 Verdeling van het aantal cases (absoluut en procentueel) over cohorten en datasets
Cohort 1880-'81
Cohort 1920
Totaal
Absoluut
Percentage
Absoluut
Percentage
Absoluut
Percentage
HSN huwelijksakten
768
41,4
1.086
58,6
1.854
100,0
Percentage
65,0
70,1
Vhmo huwelijksakten
414
47,2
464
52,9
878
100,0
Percentage
35,0
29,9
Totaal
1.182
43,3
1.550
56,7
2.732
100,0
Percentage
100,0
100,0
100,0
Bronnen: HSN release HVL 2006.01; HSN release akten 2007.01. richard l. zijdeman en kees mandemakers richard l. zijdeman en kees mandemakers richard l. zijdeman en kees mandemakers de waarden 0 tot en met 3 toe te kennen aan de verschillende onderwijstypen
wordt een hiërarchisch onderscheid gemaakt tussen de onderwijsniveaus. Om
de hiërarchie te bepalen waren we idealiter uit gegaan van waarden gebaseerd
op de werkelijke deelname aan het onderwijs, zoals behaald diploma of het aan-
tal jaren genoten opleiding. Deze informatie is echter niet voorhanden. Tabel 1
geeft een overzicht van hoe de hier gebruikte dataset per cohort is samengesteld. Tabel 2 geeft de beschrijvende statistieken van de variabelen weer voor het totaal
van beide datasets. 161 Methoden Om onze hypothesen te toetsen maken we gebruik van padanalyse. Bij padanaly-
se wordt er een causale relatie tussen de variabelen uit het model verondersteld. Figuur 1 geeft deze veronderstellingen weer. De beroepsstatus van de vader heeft
een direct effect op de beroepsstatus van de zoon. Er is echter ook sprake van
een indirect verband tussen beroepsstatus van vader en zoon. De beroepsstatus
van de zoon wordt namelijk ook bepaald door zijn opleiding en die wordt op
zijn beurt beïnvloed door de beroepsstatus van de vader. Deze verbanden worden
weergegeven in de volgende vergelijkingen Y1 = a1 + b1 * X1 + e1
(1)
Y2 = a2 + b2 * X1 + b3 * Y1+ e2 (2) Y1 = a1 + b1 * X1 + e1
(1)
Y2 = a2 + b2 * X1 + b3 * Y1+ e2 (2) waarbij X1 gelijk is aan vaders beroepsstatus, Y1 gelijk is aan de opleiding van de
zoon en Y2 gelijk is aan beroepsstatus van de zoon. Aangezien ons model recur-
sief is (de causale relaties lopen slechts in een richting), kunnen we vergelijking
(1) en (2) apart schatten met behulp van OLS-regressie onder de aanname dat
de residuen in de regressievergelijkingen van de onderliggende populatie statis-
tisch onafhankelijk zijn (Retherford & Choe, 1993, p. 95). waarbij X1 gelijk is aan vaders beroepsstatus, Y1 gelijk is aan de opleiding van de
zoon en Y2 gelijk is aan beroepsstatus van de zoon. Aangezien ons model recur-
sief is (de causale relaties lopen slechts in een richting), kunnen we vergelijking
(1) en (2) apart schatten met behulp van OLS-regressie onder de aanname dat
de residuen in de regressievergelijkingen van de onderliggende populatie statis-
tisch onafhankelijk zijn (Retherford & Choe, 1993, p. 95). We kunnen echter geen gebruikmaken van standaard OLS-regressieanaly bel 2 Beschrijvende statistieken van de variabelen voor het totaal van de HSN- en vhmo-
dataset (N = 2732) Tabel 2 Beschrijvende statistieken van de variabelen voor het totaal van de HSN- en vhmo-
dataset (N = 2732)
Variabele
Gemiddelde
Std. Dev. Min. Max. Zoons beroepsstatus
59,306
18,316
10,6
99
Vaders beroepsstatus
56,313
16,194
10,6
99
Zoons opleiding
0,616
0,979
0
3
Cohort 1920
0,567
0
1
Bronnen: HSN release HVL 2006.01; HSN release akten 2007.01. Bronnen: HSN release HVL 2006.01; HSN release akten 2007.01. Bronnen: HSN release HVL 2006.01; HSN release akten 2007.01. de rol van het gymnasiaal en middelbaar onderwijs de rol van het gymnasiaal en middelbaar onderwijs doordat we met twee verschillende databestanden werken. De dataset met vhmo-
leerlingen bevat wel informatie over opleiding, maar is alleen representatief voor
leerlingen van het vhmo. De HSN-dataset bevat weliswaar geen informatie over
opleiding, maar is wel nationaal representatief. Dat betekent dat de HSN-data-
set ook bruidegoms omvat die voorbereidend hoger en middelbaar onderwijs
genoten hebben. Om één databestand te verkrijgen waarin zowel mensen met
en zonder opleiding zijn opgenomen, schatten we aan de hand van de verdeling
van beroepsstatus van vaders enerzijds en de kans die zonen hadden om naar
het vhmo te gaan anderzijds, welk deel van de HSN-populatie geen onderwijs
genoten heeft. Om deze werkwijze te verduidelijken geven we een voorbeeld. Tabel 3 geeft een fictieve verdeling weer van beroepsstatus van de vader over drie
categorieën: laag, midden en hoog voor het bestand met vhmo-leerlingen en het
HSN-bestand. Verder wordt in dit voorbeeld gesteld dat 5 procent van de jongens
naar het vhmo gaat. 162 Om het percentage zonen zonder vhmo-opleiding van vaders met een lage
beroepsstatus te schatten gaan we als volgt te werk. In totaal heeft 55 procent
van de zonen een vader met een lage beroepsstatus. Van alle zonen heeft vijf
procent een vhmo-opleiding en daarvan heeft tien procent een vader met een
lage beroepsstatus. Het gaat er nu om de verdeling naar sociale status te bereke-
nen van de 95 procent van de vaders van wie de kinderen geen vhmo volgden. In
het geval van cel A in tabel 3, is dit gelijk aan (55 – 10*0,05) / 0,95 = 57,4 procent. Voor de cellen B en C zijn deze percentages respectievelijk 30,5 procent en 12,1
procent. Bovenstaand voorbeeld geeft een indruk van de schattingsmethode. Bij de
daadwerkelijke schatting gebruiken we voor 1880 en 1920 aparte percentages
voor deelname aan het vhmo. In 1880 ging 4,1 procent van alle jongens naar een
eerste klas vhmo, in 1920 was dit 8,1 procent (Mandemakers 1996a, tabel 14.15,
p. 593). Daarnaast gebruiken we in plaats van een verdeling in drie categorieën
een continue variabele voor de beroepsstatus van vaders: de HIS-CAM-score. Met
behulp van formules voor conditionele momenten (zie bijvoorbeeld Rao, 1973)
wordt aan de hand van het gemiddelde en de standaarddeviatie van de beroeps-
status van vaders deelname aan de verschillende typen vhmo in de HSN-popula-
tie geschat. Resultaten De sociale achtergrond van de HSN- en vhmo-cohorten 1880-'81 en 1920 De sociale achtergrond van de HSN- en vhmo-cohorten 1880-'81 en 1920
Alvorens de hypothesen te toetsen, kijken we eerst naar de verdeling van de soci-
ale achtergrond van de gehele populatie (HSN) en de vhmo-leerlingen afzonder-
lijk, voor de cohorten 1880-’81 en 1920. Figuur 3 geeft de verdeling van beroeps-
status van vaders voor de gehele populatie weer. De figuur laat zien dat de
spreiding van vaders beroepsstatus voor het cohort 1920 groter is dan voor het
cohort 1880-’81. Een Levenes variantietoets geeft aan dat dit verschil significant
is (f = 0,750; p = 0,000). Figuur 3 laat ook zien dat de gemiddelde beroepsstatus
van vaders van het cohort 1920 (51,5) hoger is dan voor vaders van het cohort
1880-’81 (47,7). Een student t-toets voor groepen met ongelijke varianties wijst
uit dat ook dit verschil in gemiddelde beroepsstatus significant is (t = –7,122;
p = 0,000). Ten opzichte van het cohort 1880-’81 is de sociale achtergrond van
zonen uit het cohort 1920 dus gemiddeld hoger, maar ook zijn de verschillen in
sociale achtergrond in het cohort 1920 groter. g
g
Figuur 4 laat verschillen in sociale achtergrond van vhmo-leerlingen zien
tussen het cohort 1880-’81 en het cohort 1920. Bij de vhmo-leerlingen is de gemid-
delde beroepsstatus van vaders afgenomen, dit in tegenstelling tot de bevindin-
gen voor de gehele populatie. De beroepsstatus van vaders van leerlingen uit het
cohort 1880-’81 bedroeg namelijk 71,6, terwijl de beroepsstatus van vaders van
het cohort 1920 significant lager is: 68,2 (t = 3,069; p = 0,001). Komt dit doordat er
in 1920 relatief meer leerlingen met een lagere sociale achtergrond deelnemen
aan het vhmo? In figuur 4 valt op dat het aandeel vaders met een extreem hoge
HIS-CAM-score (>90) relatief gezien is afgenomen. Een Levenes variantietoets laat
ook zien dat de spreiding in de beroepsstatus van vaders in 1920 significant klei-
ner is dan in 1880-’81 (f = 1,411; p = 0,000). De groei van de instroom van het vhmo
tussen 1880 en 1920 (met 400 procent) heeft geleid tot een enorme toename van
de middengroepen en slechts tot een kleine groei van de leerlingen uit de hogere
lagen. Die gingen immers in 1880 al voor een groot deel naar het vhmo. Het aan-
deel leerlingen van wie de vader een HIS-CAM-score lager dan 40 heeft is echter
niet toegenomen. de rol van het gymnasiaal en middelbaar onderwijs Dit resulteert voor beide cohorten in variantie-covariantie-matrices
van de drie variabelen: beroepsstatus vader, beroepsstatus zoon en opleiding
zoon. Daarmee kunnen vervolgens de regressiemodellen worden geschat. Omdat Tabel 3 Verdeling van beroepsstatus van de vader in drie fictieve datasets
Laag
Midden
Hoog
Vhmo (5% populatie)
10
20
70
HSN (100% populatie)
55
30
15
HSN (95% populatie)
57,4 [A]
30,5 [B]
12,1 [C] el 3 Verdeling van beroepsstatus van de vader in drie fictieve datasets richard l. zijdeman en kees mandemakers de covariantie-matrices echter niet geschat zijn op de gebruikelijke manier, zijn
de standaardfouten van de regressiecoëfficiënten niet bruikbaar. We passen
daarom een gestratificeerde bootstrap-methode toe. Met behulp van deze techniek
worden standaardfouten en betrouwbaarheidsintervallen geschat door herhaald
steekproeven te trekken uit de geobserveerde populatie.5 163 Resultaten Een zoon met een arbeidersachtergrond (40 tot 50 procent van
de samenleving) had in 1920 een kans van slechts 6 op 100 om binnen te komen
bij het vhmo (Mandemakers 1996b, 249-252). De afname in het gemiddelde en
de vermindering van de spreiding wordt dus veroorzaakt door een relatieve toe-
name van het aantal leerlingen in de middengroepen. de rol van het gymnasiaal en middelbaar onderwijs Figuur 3 Sociale status (HIS-CAM) van vaders van HSN-onderzoekspersonen voor cohort
1880-’81 en 1920 (N = 1854) Figuur 3 Sociale status (HIS-CAM) van vaders van HSN-onderzoekspersonen voor cohort
1880-’81 en 1920 (N = 1854) Figuur 3 Sociale status (HIS-CAM) van vaders van HSN-onderzoekspersonen voor cohort
1880-’81 en 1920 (N = 1854)
Bron: HSN release akten 2007 01
0
10
20
30
0
10
20
30
40
50
60
70
80
90
99
1880-’81
1920
Percentage vaders met betreffende status score (HIS-CAM)
Normaal verdeling status score vaders (HIS-CAM)
Procenten
HIS-CAM
0
10
20
30
0
10
20
30
40
50
60
70
80
90
99 Figuur 3 Sociale status (HIS-CAM) van vaders van HSN-onderzoekspersonen voor cohort
1880-’81 en 1920 (N = 1854)
0
10
20
30
0
10
20
30
40
50
60
70
80
90
99
1880-’81
1920
Procenten
HIS-CAM
0
10
20
30
0
10
20
30
40
50
60
70
80
90
99 164 Procenten Percentage vaders met betreffende status score (HIS-CAM)
Normaal verdeling status score vaders (HIS-CAM) Bron: HSN release akten 2007.01. Toetsing van de hypothesen over het statusverwervingsmodel
Alle geformuleerde hypothesen hebben betrekking op veranderingen in het sta-
tusverwervingsmodel tussen 1880-’81 en 1920. De moderniseringstheorie en de
statusbehoud-theorie verwachten beide dat de associatie tussen beroepsstatus
van bruidegoms en hun vaders afneemt (hypothese 1), terwijl de invloed van
de opleiding van bruidegoms op hun eigen beroepsstatus toeneemt (respectie-
velijk hypothese 3 en 5). Deze hypothesen zullen we dan ook eenzijdig toetsen. Wat de associatie tussen beroepsstatus van de vaders en het opleidingsniveau
van de bruidegoms betreft, verwacht de moderniseringstheorie een afname
(hypothese 2), terwijl de statusbehoud-theorie een toename in deze associatie
verwacht (hypothese 4). Dit verband toetsen we daarom tweezijdig. Tabel 4 geeft
de resultaten van de toetsingen van de hypothesen. Tevens geeft de tabel weer
hoe de verschillende effecten corresponderen met het statusverwervingsmodel
in figuur 1. richard l. zijdeman en kees mandemakers Resultaten Hypothese 1 stelt dat het directe effect van vaders beroepsstatus op dat van
de zoon (pijl A in figuur 1) over de tijd is afgenomen. Deze hypothese wordt
bevestigd. Voor 1880-’81 geldt dat wanneer de beroepsstatus van de vader met
1 standaarddeviatie toeneemt, de beroepsstatus van de bruidegom 0,574 stan-
daarddeviatie toeneemt. Voor 1920 is deze directe invloed van de beroepssta-
tus van de vader nog maar 0,479: 1,2 keer zo klein als voor het cohort 1880-’81
(peenzijdig = 0,025). richard l. zijdeman en kees mandemakers Figuur 4 Sociale status (HIS-CAM) van vaders van vhmo-leerlingen voor cohort 1880-’81 en
1920 (N = 878)
Bron: HSN release HVL 2006.01. 1880-’81
1920
Percentage vaders met betreffende status score (HIS-CAM)
Normaal verdeling status score vaders (HIS-CAM)
Procenten
HIS-CAM
0
10
20
30
0
10
20
30
40
50
60
70
80
90
99
0
10
20
30
0
10
20
30
40
50
60
70
80
90
99 Figuur 4 Sociale status (HIS-CAM) van vaders van vhmo-leerlingen voor cohort 1880-’81
1920 (N = 878)
1880-’81
1920
Procenten
HIS-CAM
0
10
20
30
0
10
20
30
40
50
60
70
80
90
99
0
10
20
30
0
10
20
30
40
50
60
70
80
90
99 165 Procenten Percentage vaders met betreffende status score (HIS-CAM)
Normaal verdeling status score vaders (HIS-CAM) Bron: HSN release HVL 2006.01. Zowel de moderniseringstheorie als de statusbehoud-theorie veronderstelde
dat de associatie tussen de opleiding van de bruidegom en zijn beroepsstatus
toeneemt (pijl C in figuur 1). Hoewel de associatie voor het cohort 1920 inder-
daad 1,3 keer zo groot is als voor het cohort 1880-’81, is deze toename niet signi-
ficant (peenzijdig = 0,074). Hypothesen 3 en 5 worden daarmee niet bevestigd. eenzijdig
De moderniseringstheorie en de statusbehoud-theorie verschillen voorname-
lijk in hun verwachting over veranderingen in de associatie tussen beroepssta-
tus van vaders en opleidingsniveau van bruidegoms (pijl B in figuur 1). Volgens
de moderniseringstheorie zou deze associatie afnemen (hypothese 2), terwijl
de statusbehoud-theorie juist een toename voorspelt (hypothese 4). Tabel 4 laat
zien dat beide hypothesen verworpen worden: hoewel er sprake is van een kleine
afname van de invloed van vaders beroepsstatus op de opleiding van de bruide-
gom, is het verschil niet significant (p = 0,218). Resultaten Naast de afzonderlijke paden in het statusverwervingsmodel (figuur 1)
geeft tabel 4 ook de veranderingen in het indirecte en in het totale effect van de
beroepsstatus van de vaders weer. Het indirecte effect dat via de opleiding van de
bruidegom loopt (pijl B en C in figuur 1) is voor het cohort 1920 iets groter (16,7
%). De toename in het indirecte effect is echter niet significant (p = 0,455). Het
totale effect van vaders beroepsstatus, dat wil zeggen het directe en indirecte
effect van vaders beroepsstatus op dat van de bruidegom, is wel significant afge- de rol van het gymnasiaal en middelbaar onderwijs Tabel 4 Uitkomsten voor het eenvoudige statusverwervingsmodel in 1880-’81 en 1920a Tabel 4 Uitkomsten voor het eenvoudige statusverwervingsmodel in 1880-’81 en 1920a
Cohort 1880-'81
Cohort 1920
H0:1880’-81 = 1920 Figuur 1
Coëf. Boot-
strapped
std. fout
Coëf. Boot-
strapped
std. fout
peenzijdig
ptweezijdig
Opleiding bruidegom
Vader
0,261
0,017
0,234
0,014
0,218
Pijl B
Beroepsstatus bruidegom
Vader (direct)
0,574
0,039
0,479
0,030
0,025
0,049
Pijl A
Vader (indirect)
0,030
0,026
0,035
0,004
0,455
Pijl B
en C
Vader (totaal)
0,604
0,005
0,514
0,027
0,042
Pijl A +
(B en C)
Opleiding bruidegom
0,116
0,019
0,150
0,015
0,074
0,147
Pijl C
n = 1182
n = 1550
a Het betreft gestandaardiseerde regressiecoëfficiënten, bootstrapped standaardfouten en p-waarden. Bronnen: HSN release HVL 2006.01; HSN release akten 2007.01. 166 nomen (p = 0,042). De totale associatie tussen de beroepsstatus van de bruidegom
en zijn vader was voor het cohort 1920 14,9 procent minder dan voor het cohort
1880-'81. nomen (p = 0,042). De totale associatie tussen de beroepsstatus van de bruidegom
en zijn vader was voor het cohort 1920 14,9 procent minder dan voor het cohort
1880-'81. Conclusie en discussie Ondanks krachtig geformuleerde hypothesen over de invloed van industrialise-
ring en onderwijsexpansie op het statusverwervingsproces zijn empirische toet-
singen van deze hypothesen in Nederland in de negentiende en het begin van
de twintigste eeuw schaars. Daarnaast beperken de enkele empirische studies
die voorhanden zijn zich veelal tot de relatie tussen beroepsstatus van vader en
zoon, terwijl tegenstrijdigheden tussen theorieën vaak betrekking hebben op de
rol van onderwijs. Door twee unieke datasets met elkaar te combineren geeft
deze studie voor het eerst inzicht in het statusverwervingsproces inclusief het
belang van onderwijs voor Nederland aan het eind van de negentiende en het
begin van de twintigste eeuw. Hoewel door de expansie van het onderwijs het aantal leerlingen van het
vhmo toenam tussen 1880 en 1920, is het de vraag of de verdeling van leerlingen
naar sociale achtergrond minder scheef is geworden. Onze data wijzen uit dat
tussen 1880 en 1920 het aandeel van leerlingen met een zeer hoge sociale ach-
tergrond afnam, door een toename van leerlingen met een gemiddeld tot hoge
sociale achtergrond, zoals kinderen van kantoorbedienden, middelbare techni- richard l. zijdeman en kees mandemakers ci, onderwijzers en andere nieuwe middengroepen. Toegang tot het vhmo voor
leerlingen met een arbeidersachtergrond was ook in 1920 nog zeer beperkt. Voor
zonen van vaders met lage status bleef het vhmo in 1920, bijna 60 jaar na zijn
introductie, vrijwel onbereikbaar. 167 Toch neemt het verband tussen beroepsstatus van vaders en zonen in de
periode 1880-1920 af. Dit is het gevolg van een daling van de directe invloed van
vaders beroepsstatus op dat van zijn zoon. Deze bevinding sluit aan op de studie
van Boonstra (1993) die voor de gehele negentiende eeuw een belangrijke daling
vond in de samenhang tussen de sociale status van de vader en die van de zoon. Boonstra vond tevens dat de invloed van alfabetisme, het gevolg van minimaal
enkele jaren opleiding, op eigen beroepsstatus na 1850 afnam (p. 207). Het is ech-
ter onduidelijk of de invloed van onderwijs op de eigen beroepsstatus daadwer-
kelijk afnam. De door Boonstra gevonden afname zou namelijk verklaard kun-
nen worden door de sterke afname van het aantal analfabeten. Hierdoor boette
alfabetisme als onderscheidend kenmerk voor beroepsstatus aan belang in. Op
basis van de resultaten uit deze studie lijkt dit laatste inderdaad het geval te zijn
geweest. Tussen 1880-’81 en 1920 vinden we namelijk geen verandering in de
invloed van opleiding op beroepsstatus. Conclusie en discussie Daarnaast laten de resultaten ook geen
verandering zien in het verband tussen beroepsstatus van de vader en opleiding
van de zoon. Onze bevindingen bieden slechts geringe ondersteuning voor de modernise-
ringstheorie en voor de statusbehoud-theorie. De afname in het directe verband
tussen beroepsstatus van de vader en die van de zoon wordt niet gecompenseerd
door extra investering in opleiding, zoals verondersteld door de statusbehoud-
theorie. De associatie tussen achtergrond en opleiding neemt echter ook niet af
zoals verwacht door de moderniseringstheorie. Daarnaast neemt het belang van
opleiding voor beroepsstatus ook niet toe, zoals beide theorieën verwachten. Verder onderzoek moet uitwijzen of het algemene beeld dat deze studie
schetst recht doet aan verschillen op het individuele niveau en aan verschillen
op het regionale niveau. Helaas beschikten we in deze studie nauwelijks over
kenmerken anders dan opleiding en beroep. Wanneer bijvoorbeeld het status-
verwervingsproces gedifferentieerd zou kunnen worden naar verschillende reli-
gieuze groepen zou kunnen blijken of de door de theorieën verwachte effecten
al wel optraden binnen bepaalde geloofsgemeenschappen. Daarnaast zouden er
regionale verschillen in het statusverwervingsproces kunnen zijn die door onze
analyses onopgemerkt zijn gebleven. Industrialisering en onderwijsexpansie
drongen niet overal op hetzelfde tijdstip en ook niet in hetzelfde tempo door. Of
dergelijke verschillen in de toekomst gevonden worden zal staan of vallen met
de kwaliteit en kwantiteit van te verzamelen historische data. de rol van het gymnasiaal en middelbaar onderwijs Noten 1. Wij willen graag Jeroen Weesie bedanken voor zijn hulp bij de analyses. Commentaren
van de editors en die van Jan Kok op een eerdere versie van dit hoofdstuk vormen een
belangrijke bijdrage. Ook zijn we dankbaar voor de commentaren die we hebben ontvan-
gen bij presentaties van dit hoofdstuk op het COST Action A-34 / ESTER Advanced Seminar
2007 en het Social Stratification Research Seminar 2007. Deze studie is onderdeel van het
promotieproject: ‘Status attainment during industrialization, life courses in local con-
text’. Het project is mede mogelijk gemaakt door financiering van de Nederlandse Organi-
satie voor Wetenschappelijk Onderzoek (NWO) (MAGW Open Competitie 400-05-054). Het
op grote schaal lokaliseren van de huwelijksakten van vhmo-leerlingen was niet haalbaar
geweest zonder het bestaan van de digitale archieven in de appendix. 168 2. Dit gebeurde aan de hand van aantekeningen over de levenslopen van deze leerlingen die
zich bevonden in de dossiers die per vhmo-leerling werden aangelegd, zie Mandemakers
(1992). 2. Dit gebeurde aan de hand van aantekeningen over de levenslopen van deze leerlingen die
zich bevonden in de dossiers die per vhmo-leerling werden aangelegd, zie Mandemakers
(1992). (
)
3. De digitale archieven die werden gebruikt zijn weergegeven in de appendix. (
)
3. De digitale archieven die werden gebruikt zijn weergegeven in de appendix. 4. Het databestand is ondergebracht bij de Historische Steekproef Nederlandse bevolking (HSN) als
Dataset Huwelijksakten VHMO-leerlingen 1880-1881 en 1920 (HVL), release 2006.01 (Zijdeman,
2006). 4. Het databestand is ondergebracht bij de Historische Steekproef Nederlandse bevolking (HSN) als
Dataset Huwelijksakten VHMO-leerlingen 1880-1881 en 1920 (HVL), release 2006.01 (Zijdeman,
2006). 2006). 5. Een Stata-script waarmee deze schattingsmethode is uitgewerkt is beschikbaar. )
5. Een Stata-script waarmee deze schattingsmethode is uitgewerkt is beschikbaar. Literatuur Blau, P.M. & O.D. Duncan (1967). The American occupational structure. New York:
Wiley. Boekholt, P.Th.F.M. & E.P. de Booy (1987). Geschiedenis van de school in Nederland. Vanaf de middeleeuwen tot aan de huidige tijd. Assen en Maastricht: Van Gorcum. Boonstra, O.W.A. (1993). De waardij van eene vroege opleiding. Een onderzoek naar de
implicaties van het alfabetisme op het leven van inwoners van Eindhoven en omlig-
gende gemeenten, 1800-1920. Wageningen: Afdeling Agrarische Geschiedenis
Landbouwuniversiteit. Boonstra, O.W.A. (1995). Het einde van het analfabetisme. In C.A. Mandemakers
& O.W.A. Boonstra (red.), De levensloop van de Utrechtse bevolking in de negentiende
eeuw (pp. 68-85). Assen: Van Gorcum. Boonstra, O.W.A. & C.A. Mandemakers (1995). “Ieder is het kind zijner eigene wer-
ken”. Sociale mobiliteit in Nederland in de achttiende en negentiende eeuw. In J. Dronkers & W.C. Ultee (red.), Verschuivende ongelijkheid in Nederland (pp. 125-141). Assen: Van Gorcum. Bourdieu, P. & J.C. Passeron (1977[1970]), Reproduction, Londen: Sage. Bras, H. & J. Kok (2005). “They live in indifference together”: Marriage mobility
in Zeeland, The Netherlands, 1796-1922. In M.H.D. van Leeuwen, I. Maas & A. Miles (red.), Marriage choices and class boundaries: Social endogamy in history (pp. 247-274). Cambridge: Cambridge University Press. CBS (1959). Zestig jaren statistiek in tijdreeksen 1899-1959. Zeist: De Haan. llins, R. (1971). Functional and conflict theories of educational stratification. Collins, R. (1971). Functional and conflict theories of educational stratification. American Sociological Review, 36, 1002-1019. American Sociological Review, 36, 1002-1019. American Sociological Review, 36, 1002-1019. richard l. zijdeman en kees mandemakers Collins, R. (1979). The credential society, New York: Academic Press. Delger, H. & J. Kok (1998). Bridegrooms and biases. Historical Methods, 31, 113-12 Van Dijk, H. & C.A. Mandemakers (1985). Secondary education and social mobi-
lity at the turn of the century. History of Education, 14(3), 199-226. 169 Dronkers, J. & P.M. de Graaf (1995). Ouders en het onderwijs van hun kinderen. In J. Dronkers & W.C. Ultee (red.), Verschuivende ongelijkheid in Nederland (pp. 46-67). Assen: Van Gorcum. Ganzeboom, H.B.G. & R. Luijkx (1995). Intergenerationele beroepsmobiliteit in
Nederland: Patronen en historische veranderingen. In J. Dronkers & W.C. Ultee
(red.), Verschuivende ongelijkheid in Nederland (pp. 14-45). Assen: Van Gorcum. Ganzeboom, H.B.G., D.J. Treiman & W.C. Ultee (1991). Comparative intergenera-
tional stratification research: Three generations and beyond. Annual Review of
Sociology, 17, 277-302. Goodman, L.A. (1979). Simple models for the analysis of association in cross-clas-
sifications having ordered categories. Literatuur Journal of the American Statistical Associa-
tion, 74, 537-552. Grusky, D.B. (1983). Industrialization and the status attainment process: The the-
sis of industrialism reconsidered. American Sociological Review, 48, 494-506. Harrigan, P.J. (1980). Mobility, elites, and education in French society of the second empi-
re. Waterloo Ontario: Wilfrid Laurier University Press. International Standard Classification of Occupations: Revised edition 1968. (1969). Gene-
va: International Labour Office. Kaelble, H. (1983). Soziale Mobilität und Chancengleichheit im 19. und 20. Jahrhundert. Deutschland im internationalenVergleich. Göttingen: Vandenhoeck & Ruprecht. Kerr, C., J.T. Dunlop, F. Harbison & C. A. Myers (1973 [1960]). Industrialism and in-
dustrial man. Middlesex: Pelican. Kuznets, S. (1957). Quantitative aspects of the economic growth of nations: II. Industrial distribution of national product and labor force. Economic Develop-
ment and Cultural Change, 5, 1-111. Lambert, P.S., R.L. Zijdeman, I. Maas, K. Prandy & M.H.D. van Leeuwen (2006). Testing the universality of historical occupational stratification structures
across time and space. ISA RC 28 Social Stratification and Mobility spring meeting. Radboud University, Nijmegen. Leeuwen, M.H.D. van & I. Maas (1995). Sociale mobiliteit in de steden en op het
platteland. In C.A. Mandemakers & O.W.A. Boonstra (red.), De levensloop van de
Utrechtse bevolking in de 19e eeuw (pp. 103-128). Assen: Van Gorcum. Leeuwen, M.H.D. van & I. Maas (1997). Social mobility in a Dutch province. Journal
of Social History, 30, 619-644. Leeuwen, M.H.D. van & I. Maas (2007). Economische specialisering en verande-
rende sociale verhoudingen in de 19e en 20e eeuw. Een studie op basis van
de Nederlandse volkstellingen en huwelijksakten. In O.W.A. Boonstra, P.K. Doorn, M.P.M. van Horik, J.G.S.J. van Maarseveen & J. Oudhof (red.), Twee eeu-
wen Nederland geteld. Onderzoek met de digitale Volks- Beroeps- en Woningtellingen
1795-2001 (pp. 181-205). Den Haag: DANS en CBS. de rol van het gymnasiaal en middelbaar onderwijs Leeuwen, M.H.D. van, I. Maas & A. Miles (2002). HISCO – Historical Occupational
Standard Classification of Occupations. Leuven: Leuven University Press. Lundgreen, P., M. Kraul & K. Ditt (1988). Bildingschancen und soziale Mobilität in der
städtischen Gesellschaft des 19. Jahrhunderst. Göttingen: Vandenhoeck & Rup-
recht. 170 Maas, I., P.S. Lambert, R.L. Zijdeman, K. Prandy & M.H.D. van Leeuwen (2006). HIS-
CAM, the derivation and implementation of a historical occupational stratifi-
cation scale. Sixth European Social Science History Conference. RAI, Amsterdam. Mandemakers, K. (1992). Education and social mobility: Organizing and storing
an historical survey. In P. Doorn, C. Kluts & E. Literatuur A test of the influence of industrialisa-
tion, mass communication and urbanisation in 117 municipalities. Internatio-
nal Journal of Sociology and Social Policy, 28, 204-216. Zijdeman, R.L. (2008). Intergenerational transfer of occupational status in 19th
century Zeeland, The Netherlands. A test of the influence of industrialisa-
tion, mass communication and urbanisation in 117 municipalities. Internatio-
nal Journal of Sociology and Social Policy, 28, 204-216. Zijdeman, R.L. (2008). Intergenerational transfer of occupational status in 19th
century Zeeland, The Netherlands. A test of the influence of industrialisa-
tion, mass communication and urbanisation in 117 municipalities. Internatio-
nal Journal of Sociology and Social Policy, 28, 204-216. Literatuur Leenarts (red.), Data, computers
and the past. Proceedings of the conference Archiving and disseminating historical
machine readable data (pp. 149-161). Hilversum: Verloren. Cahier VGI 5. Mandemakers, K. (1996a). Gymnasiaal en middelbaar onderwijs. Ontwikkeling, struc-
tuur, sociale achtergrond en schoolprestaties, Nederland, ca. 1800-1968, band II. Rot-
terdam: Erasmus Universiteit. Mandemakers, K. (1996b). HBS en gymnasium. Ontwikkeling, structuur, sociale achter-
grond en schoolprestaties, Nederland, circa 1800-1968 (Vol. 24). Amsterdam: Stich-
ting Beheer IISG. Mandemakers, K. (1999). Onderwijsdeelname, 1870-1990. In R. van der Bie & P. Dehing (red.), Nationaal goed. Feiten en cijfers over onze samenleving (ca.) 1800-1999
(pp. 179-197). Amsterdam: Stichting Beheer IISG. Mandemakers, K. (2001). De sociale structuur in Nederland rond 1900. De samen-
leving in het perspectief van de modernisering 1850-1990. In: J.G.S.J. van Maar-
seveen en P.K. Doorn (red.), Nederland een eeuw geleden geteld. Een terugblik op de
samenleving rond 1900 (pp. 185-207). Amsterdam: (Stichting Beheer IISG). Parsons, T., & E.A. Shils (red.) (1951). Toward a general theory of action. Cambridge:
Harvard University Press. Ploeg, S.W. van der (1993). The expansion of secondary and tertiary education in the
Netherlands. Nijmegen: ITS. Poppel, F. van, C. Monden & K. Mandemakers (2008). Marriage timing over the
generations. Human Nature, 19, 7-22. Rao, C.R. (1973). Linear statistical inference and its applications. New York: Wiley. Retherford, R.D. & M.K. Choe (1993). Statistical models for causal analysis. New York:
John Wiley & Sons, Inc. Rijken, S. (1999). Educational expansion and status attainment. A cross-national and
over-time comparison. Utrecht: Utrecht University. Ringer, F.K. (1979). Education and society in modern Europe. Bloomington and Lon-
don: Indiana University Press. Treiman, D.J. (1970). Industrialization and social stratification. Social Inquiry, 40,
207-234. Van Zanden, J.L. & R.T. Griffiths (1989). Economische geschiedenis van Nederland in de
20 eeuw. Utrecht: Spectrum. Zanden, J.L. van & A. van Riel (2000). Nederland 1780-1914. Staat, instituties en econo-
mische ontwikkeling. Z.pl.: Balans. richard l. zijdeman en kees mandemakers Zijdeman, R.L. (2006). Historische Steekproef Nederlandse bevolking (HSN). Data-
set huwelijksakten VHMO-leerlingen 1880-’81 en 1920 (HVL), release 2006.01. Zijdeman, R.L. (2007). “Nieuwe data, ‘nieuwe’ methode?” Een sociologisch his-
torische toepassing van multiniveau-analyse. In O.W.A. Boonstra, P.K. Doorn,
M.P.M. van Horik, J.G.S.J. van Maarseveen en J. Oudhof (red.), Twee eeuwen Neder-
land geteld. Onderzoek met de digitale Volks- Beroeps- en Woningtellingen 1795-2001
(pp. 441-454). Den Haag: DANS en CBS. 171 Zijdeman, R.L. (2008). Intergenerational transfer of occupational status in 19th
century Zeeland, The Netherlands. Appendix Gemeente-, streek-, en regionale archieven die geraadpleegd zijn bij
het lokaliseren van de huwelijksakten van de vhmo-leerlingen* de rol van het gymnasiaal en middelbaar onderwijs Regionaal Archief Leiden
http://www.leidenarchief.nl/
Streekarchief Voorne-Putten en Rozenburg
http://www.streekarchiefvpr.nl/
Toegang Op Personen (TOP)
http://top.archiefplein.nl/websitepubliek/
Tresoar
http://www.tresoar.nl/
Virtuele Studiezaal Gemeente Den Haag
http://193.172.27.173/virtuelestudiezaal/selectiebron.aspx
Historisch Centrum Overijssel
http://www.historischcentrumoverijssel.nl/hcoroot
* webadressen per 25 april 2008 172 * webadressen per 25 april 2008 Van een dubbeltje naar een kwartje?
Beroepsloopbanen van mannen en vrouwen in Nederland
tussen 1865 en 1940 Ineke Maas en Marco H.D. van Leeuwen1 Ineke Maas en Marco H.D. van Leeuwen1 Inleiding Hoe zagen de beroepsloopbanen van Nederlandse mannen en vrouwen er in het
verleden uit? Maakten zij carrière, in de zin van opwaartse mobiliteit in de loop
van hun leven? Was neerwaartse mobiliteit een frequente ervaring of was er een
gebrek aan mobiliteit? Daar weten we nog nauwelijks iets vanaf. Dat komt in
belangrijke mate door het gebrek aan makkelijk toegankelijke bronnen (Brown,
Van Leeuwen & Mitch, 2004). Surveys reiken zelden verder terug dan de Tweede
Wereldoorlog. Tot nu toe is er daarom vooral met bronnen gewerkt die maar
een klein gedeelte van beroepsloopbanen in kaart brengen, bijvoorbeeld door
het beroep bij huwelijk te vergelijken met dat bij het huwelijk van een van de
kinderen (Van Leeuwen & Maas, 1995); of met bronnen die een klein en bijzon-
der deel van de bevolking betreffen. Voorbeelden daarvan zijn autobiografieën
(Miles, 1999), personeelsdossiers van grote bedrijven (Oosterhuis, 1992; Stovel,
Savage & Bearman, 1996) en loopbaangegevens van een specifieke beroepsgroep,
zoals dienstboden (Bras, 2002). In dit hoofdstuk willen we nu eens niet een klein
gedeelte van de loopbaan, of de gehele loopbaan van een klein gedeelte van de
beroepsbevolking onderzoeken, maar de loopbanen van een dwarsdoorsnede van
een substantieel deel van de Nederlandse bevolking en van zowel mannen als
vrouwen. We bestuderen de periode 1865-1940, decennia die zelfs door middel
van het oudste survey-onderzoek niet meer in kaart gebracht kunnen worden. Tevens zullen we aandacht besteden aan een andere kwestie. Een belang-
rijke vraag in mobiliteitsonderzoek is in hoeverre de kansen op een succesvolle
beroepscarrière afhangen van de sociale positie van het gezin van herkomst. Onderzoek naar intergenerationele mobiliteit laat een sterke samenhang zien
tussen het beroep van de vader en dat van zijn zoon, zowel tegenwoordig als in
voorgaande eeuwen (Breen, 2004; Erikson & Goldthorpe, 1992; Maas & van Leeu-
wen, 2002) Datzelfde geldt voor moeders en dochters, ofschoon onderzoek hier-
naar zeldzamer is (Korupp, 2000; Maas & Van Leeuwen, 2006). Een samenhang tussen het beroep van ouder en kind op een willekeurig moment in hun leven
kan op verschillende manieren tot stand komen. Kinderen uit hogere sociale
groepen kunnen een betere start maken en al in hun eerste beroep een relatief
hoge sociale status bezitten. Daarnaast is het mogelijk dat hun beroepsloopba-
nen zich succesvoller ontwikkelen dan die van kinderen uit lagere sociale groe-
pen. In welke mate beide processen een rol spelen is weinig onderzocht. Inleiding 174 Zowel bij het beschrijven van de beroepsloopbanen van de hele bevolking als
bij het nagaan van het effect van de sociale status van het gezin van herkomst,
is het interessant te kijken naar langetermijnontwikkelingen. Volgens de indus-
trialiseringsthese veranderden tijdens de industrialisering zowel de mate van
opwaartse en neerwaartse mobiliteit als de invloed van het gezin van herkomst
hierop (Kerr, 1960; Treiman, 1970). Mobiliteit werd steeds waarschijnlijker en de
invloed van het gezin van herkomst op de mobiliteitskansen van de kinderen
zou zijn afgenomen. De Nederlandse samenleving industrialiseerde sterk gedu-
rende de tweede helft van de negentiende en de eerste helft van de twintigste
eeuw. Bij gebrek aan bronnen is de industrialiseringsthese in deze context ech-
ter nog nauwelijks getest, en daar beogen wij verandering in te brengen. Kortom, in deze bijdrage beschrijven we de beroepscarrières van een groot
deel van de Nederlandse bevolking, mannen zowel als vrouwen, in de periode
1865 tot 1940. We onderzoeken daarbij de invloed van de beroepsstatus van de
vader op de beroepscarrière van zijn zonen en dochters en de mate waarin deze
invloed veranderde tijdens de industrialisering We stellen de volgende vragen invloed veranderde tijdens de industrialisering. We stellen de volgende vra – In welke mate kunnen de beroepsloopbanen van Nederlandse mannen
en vrouwen tussen 1865 en 1940 gekenmerkt worden als stabiel, opwaarts
mobiel of neerwaarts mobiel? Welke veranderingen treden er op in de loop
van de tijd? – In welke mate, en hoe, werden deze beroepsloopbanen beïnvloed door de
beroepsklasse van de vader? Verandert deze invloed in de loop van de tijd? – In welke mate, en hoe, werden deze beroepsloopbanen beïnvloed door de
beroepsklasse van de vader? Verandert deze invloed in de loop van de tijd? De data die verzameld zijn in het project Life Courses in Context van de Histori-
sche Steekproef Nederland (Mandemakers, 2004) geven een eerste mogelijkheid
om deze vragen voor Nederland te beantwoorden. Deze dataset bevat gegevens
over het beroep van mannen en vrouwen op meerdere tijdstippen in hun leven,
bijvoorbeeld bij verhuizingen en bij de geboorte van hun kinderen. We bestude-
ren de geboortecohorten tussen 1850 en 1882 waarvan de beroepsloopbanen zich
uitstrekten van 1865 tot aan de Tweede Wereldoorlog. Te toetsen veronderstellingen Hypothesen over de mate van beroepsmobiliteit in de tweede helft van de negentiende eeuw
Er zijn verschillende redenen om aan te nemen dat beroepsmobiliteit ook – en
misschien zelfs wel vooral – in het begin van de hier onderzochte periode veel ineke maas en marco h.d. van leeuwen voorkwam. Een groot deel van de beroepsbevolking bestond in deze tijd uit arbei-
ders en slechts weinigen van hen waren permanent verzekerd van voldoende
inkomsten. Slechts een minderheid van de arbeidskrachten had een langdurig
arbeidscontract. Binnen de groep landarbeiders lieten zich globaal drie deel-
groepen onderscheiden: inwonend vast personeel, uitwonend vast personeel en
losse arbeiders. Het ging hierbij zowel om mannen, vrouwen als kinderen (Pries-
ter, 1991). Inwonend personeel verbond zich voor een ‘relatief lange’ tijd, vaak
een jaar, voor een vast loon. Vaste uitwonende arbeiders contracteerden zich
eveneens voor een heel jaar tegen een vooraf bepaald tijdloon. Losse arbeiders
werkten als er werk was en werden per dag of karwei aangenomen. 175 Een groot deel van de arbeidskrachten was een gedeelte van het jaar werk-
loos. Vooral in de winter en tijdens een economische baisse was er een overschot
aan ongeschoolde arbeiders. Over Amsterdam schreef men: ‘Het winterseizoen
is een zorgelijk tijdvak voor de arbeidende volksklasse. En dan is er inderdaad
veelal gebrek aan werk omdat een groot gedeelte der ambachten en handnerin-
gen van dien aard zijn, dat zij alleen in de zomermaanden kunnen uitgeoefend
worden; terwijl ook de mindere levendheid van handel en scheepvaart in de
wintermaanden de vele bedrijven die aan de hoofdbronnen van onze nationale
welvaart zijn verbonden, doet stilstaan, zoodat die talrijke klasse van ambachts-
lieden, kruijers, sjouwers en dergelijke, die van de bedrijven leven die uit den
handel en de scheepvaart voortvloeyen, inderdaad zonder werk zijn.’ (Van Leeu-
wen, 1992: 55) Iedere keer dat de contracten van arbeiders afliepen en iedere keer dat arbei-
ders werkloos waren, moesten zij op zoek naar iets nieuws. Korte contracten en
seizoenswerkloosheid waren dus potentiële oorzaken van beroepsmobiliteit. Die
nieuwe baan was immers niet noodzakelijkerwijs in hetzelfde beroep. Hiervan
getuigt bijvoorbeeld een passage in de levensbeschrijving van Neel Doff, Dagen
van honger en ellende, die betrekking heeft op joodse venters in de jaren zestig
van de negentiende eeuw: ‘Ik zei aan één van hen, hoe ’t mij verwonderde, dat
ik hem broches met glazen steentjes zag verkoopen, terwijl hij de vorige week in
vijgen deed. Te toetsen veronderstellingen Hij antwoordde in drukke zinnen, dat hij elke acht dagen een ande-
re negotie nam, dat de verkoop in de wijk geen twee weken achter elkaar ging
met hetzelfde artikel, dat je met je tijd mee moest gaan en telkens wat nieuws
hebben.’ (Doff, 1974: 93) Beroepsmobiliteit na afloop van het contract of na werkloosheid zal echter
meestal hebben bestaan uit wisselingen tussen baantjes van dezelfde soort, die
ook min of meer dezelfde status hadden, omdat het hier ongeschoolde arbeiders
betrof, die moeilijk toegang kregen tot de deelmarkten voor geschoolde arbeid. Deze toegang werd ten dele geregeld door gildeachtige organisaties of was vaak
voorbehouden aan arbeiders met een gemeenschappelijke achtergrond: in
Amsterdam bijvoorbeeld in de scheepsbouw, de huizenbouw (veel Brabanders),
de bakkerijen (vrijwel uitsluitend Duitsers) en de diamantbewerking (veel joden)
(Van Leeuwen, 2000). van een dubbeltje naar een kwartje? Naast seizoenswerkloosheid en tijdelijke contracten beïnvloedden ook veran-
deringen in de vraag naar verschillende soorten arbeiders en personeel de mate
van beroepsmobiliteit. De voortschrijdende industrialisering in de onderzochte
periode was een drijvende kracht achter dergelijke veranderingen. De meest
complete bron voor de beschrijving van ontwikkelingen op de arbeidsmarkt zijn
vermoedelijk de huwelijksakten (zie voor de voor- en nadelen van deze bron Van
Leeuwen & Maas, 2007; voor een benadering gebaseerd op steekjaren zie Mande-
makers, 2001). Voor de provincie Zeeland zijn alle huwelijksakten gedigitaliseerd
in het kader van het Genliasproject. De onderstaande beschrijving is gebaseerd
op een eerder gepubliceerde analyse door ons van deze gegevens (Van Leeuwen &
Maas, 2007). Vanzelfsprekend is Zeeland niet representatief voor heel Nederland. Maar aangezien het ons hier gaat om de grote lijnen van veranderingen in de
beroepsstructuur en omdat soortgelijke gegevens voor heel Nederland nog niet
beschikbaar zijn, maken we toch gebruik van de Zeeuwse gegevens. 176 Als we kijken naar veranderingen in de verdeling van de bruidegoms over
beroepsklassen (figuur 1), is de meest in het oog springende ontwikkeling de
afname van de ongeschoolde arbeiders buiten de agrarische sector. Hun aandeel
daalde van ongeveer 40 procent van het aantal bruidegoms rond 1860 tot 13 pro-
cent rond 1920. Er is daarentegen een lichte stijging te zien van het percentage
huwende mannen dat in de primaire sector werkzaam was. De boeren en boe-
renarbeiders, waar het naast de vissers in hoofdzaak om ging, zagen het aandeel
van hun sociale lagen stijgen van ruim 30 procent rond 1860 tot zo’n 40 procent
in 1920. Te toetsen veronderstellingen De klassen van lagere managers en lagere vrije beroepen, klerken en
winkelpersoneel (sociale klassen 3, 4 en 5) groeiden van rond de 8 procent van
alle huwende mannen omstreeks de helft van de negentiende eeuw tot krap 20
procent in de periode 1916-1922. Het aandeel van de geschoolde arbeiders en
voormannen (sociale klassen 6 en 7) liet een geringe stijging zien. Op de arbeids-
markt in zijn geheel was er sprake van een toename van beroepen met een rela-
tief hoge status en een afname van beroepen onder aan de statusladder. We ver-
wachten daarom dat opwaartse mobiliteit vaker voorkwam dan neerwaartse. Hypothesen over veranderingen in de mate van beroepsmobiliteit tussen 1865 en 1940
Veranderingen in de beroepsstructuur leiden niet per se tot veranderingen in
de mate van beroepsmobiliteit. Gelijkmatige veranderingen in de beroepsstruc-
tuur kunnen tot stand komen door een constante mate van beroepsmobiliteit. Zo gaat een constant overschot aan opwaartse mobiliteit samen met een groei
van klassen met een hoge status en een constant overschot aan neerwaartse
mobiliteit met een krimp van deze klassen. Slechts indien er sprake is van een
versnelling of vertraging van de structurele veranderingen, neemt de mate van
beroepsmobiliteit toe respectievelijk af. Figuur 1 laat zien dat de veranderingen
in de beroepsstructuur, althans in Zeeland, zich min of meer constant voorde-
den gedurende de gehele onderzochte periode. Op grond van veranderingen in ineke maas en marco h.d. van leeuwen Figuur 1 Verdeling van Zeeuwse bruidegoms over de verschillende beroepsklassen (HISCLASS
naar periode Figuur 1 Verdeling van Zeeuwse bruidegoms over de verschillende beroepsklassen (HISCLASS),
naar periode Figuur 1 Verdeling van Zeeuwse bruidegoms over de verschillende beroepsklassen (HISCLASS),
naar periode
Legenda: 1 + 2 = Hogere managers en vrije beroepen; 3 + 4 + 5 = Lagere managers en vrije beroepen,
klerken en winkelpersoneel; 6 + 7 = Geschoolde arbeiders; 8 = Boeren en vissers; 9 = Laag geschoolde
arbeiders; 11 = Ongeschoolde arbeiders; 10 + 12 = Landarbeiders. Bron: Van Leeuwen & Maas, 2007. Te toetsen veronderstellingen 0
5
10
15
20
25
30
35
40
45
1856-1865
1876-1885
1896-1905
1916-1922
periode
%
1+2
3+4+5
6+7
8
9
11
10+12 Figuur 1 Verdeling van Zeeuwse bruidegoms over de verschillende beroepsklassen (HISCLASS),
naar periode
0
5
10
15
20
25
30
35
40
45
1856-1865
1876-1885
1896-1905
1916-1922
periode
%
1+2
3+4+5
6+7
8
9
11
10+12 177 Legenda: 1 + 2 = Hogere managers en vrije beroepen; 3 + 4 + 5 = Lagere managers en vrije beroepen,
klerken en winkelpersoneel; 6 + 7 = Geschoolde arbeiders; 8 = Boeren en vissers; 9 = Laag geschoolde
arbeiders; 11 = Ongeschoolde arbeiders; 10 + 12 = Landarbeiders. Bron: Van Leeuwen & Maas, 2007. de beroepsstructuur verwachten we daarom geen veranderingen in de mate van
opwaartse en neerwaartse mobiliteit. Andere ontwikkelingen op de arbeidsmarkt suggereren wel veranderingen in
de mate van beroepsmobiliteit. In de loop van de negentiende eeuw nam op het
platteland losse arbeid steeds meer toe ten koste van vast werk en prestatieloon
ten koste van tijdloon. Oorzaken daarvan waren de steeds grotere schaal van het
bedrijf, waardoor de boer niet overal meer toezicht kon houden, en het verdwij-
nen van een groot aantal taken die vroeger door vaste en inwonende arbeiders
werden gedaan. Aangezien losse contracten een potentiële bron van mobiliteit
zijn, verwachten we een toename van de beroepsmobiliteit, vooral naar beroe-
pen met ongeveer dezelfde (lage) status. Daar staat echter tegenover dat de gevoeligheid voor seizoenswerkloosheid
van een aantal sectoren in de twintigste eeuw sterk afnam, hoewel ze zeker niet
verdween. De invoering van de graanelevatoren in de Rotterdamse haven aan het
begin van de twintigste eeuw betekende dat er in de zomer maar liefst 1300 tot
1400 havenarbeiders minder nodig waren om het graan te laden en te lossen. Voor de machinale verwerking van steenkool en erts gold hetzelfde: veel steen-
kolenarbeiders verloren hun werk, maar wie overbleef kreeg vaker dan voorheen van een dubbeltje naar een kwartje? vast in plaats van los werk. Te toetsen veronderstellingen Ook in andere delen van de arbeidsmarkt vermin-
derde aan het eind van de negentiende eeuw de seizoenswerkloosheid door de
invoering van nieuwe productietechnieken, zoals in de bouw ‘terwijl vroeger het
gebruik van tras en kalk bij het metselen veroorzaakte, dat men gedurende het
jaargetijde, waarin de vorst te wachten was, geen eigenlijken buitenarbeid ver-
richten kon, is door het gebruik van de zoveel sneller verhardende cement alleen
bij strenge vorst het metselen onmogelijk geworden’ (geciteerd bij Van Leeuwen
2000, 200). De afname van de omvang van de ongeschoolde arbeidersklasse zou
samen met de verminderde seizoenswerkloosheid moeten leiden tot een afname
van de beroepsmobiliteit. Opnieuw betreft het hier vooral mobiliteit van beroe-
pen met een lage status naar soortgelijke beroepen. 178 Hypothesen over de ‘vorm’ van de beroepscarrière spreekt in zulke gevallen wel van ‘pensionering op de werkplek’ (retirement on the
job). 179 Hypothesen over de invloed van vaders beroepsklasse op het carrièreverloop van zijn
kinderen Arbeiderskinderen kwamen veelal aan werk op voorspraak van familie en vrien-
den. Dat wervingssysteem was goedkoop voor het bedrijf, terwijl het ook een
zekere garantie van kwaliteit inhield, omdat men de betrokken personeelsleden
kon aanspreken op de prestaties van hun kandidaten. Ook de spoorwegen, pos-
terijen en de belastingdienst, grote bureaucratische bedrijven die prijs stelden
op betrouwbaar en loyaal personeel dat lang in dienst bleef, rekruteerden hun
personeel op deze wijze. Een hoge functionaris bij het spoor verklaarde in 1882
zelfs dat er op termijn een ‘familie’ ontstond ‘waarvan het bestaan zich verbindt
en zamengroeit met de maatschappij zelven, eene soort van dynastie van spoor-
wegbeambten’ (Dehing, 1989, 84). Zoals eerder al gesteld waren beroepswisselingen vermoedelijk niet zeld-
zaam. Dat betekent dat de ouders steeds opnieuw een kans kregen om iets voor
hun kinderen te doen. De verschillende mechanismen die deze invloed veroor-
zaken zijn vooral van toepassing op jonge leeftijd wanneer zoons of dochters
het beroep van hun vader of moeder leerden, naar school gingen en al dan niet
door hun ouders geholpen werden bij hun huiswerk. Ook het doorgeven van een
boerderij of een winkel aan een kind of het kopen daarvan voor een ander kind
(Damsma & Kok, 2005) gebeurde eerder aan het begin dan aan het eind van de
loopbaan van het kind. Over het algemeen verwachten we daarom vooral dat
zonen en dochters van vaders uit hogere beroepsklassen al in hun eerste beroep
een hogere status hadden dan de kinderen uit lagere beroepsklassen. Daarnaast
verwachten we dat de eerste groep meer steeg tijdens de beroepsloopbaan. De invloed van het beroep van de vader als occupational broker (Miles, 1999)
betrof vooral zijn zonen. De beroepshorizon van vrouwen was veel geringer. Voor
zover zij betaald werk verrichtten, deden verreweg de meeste vrouwen dat in een
klein aantal ongeschoolde, typische vrouwenberoepen. Dat verkleinde de moge-
lijkheid van hun vader hen te helpen. Hypothesen over de ‘vorm’ van de beroepscarrière Hypothesen over de ‘vorm’ van de beroepscarrière Hypothesen over de vorm van de beroepscarrière
Er zijn redenen om te veronderstellen dat het aantal keer dat men van baan wis-
selde en of dat in op- of neerwaartse richting gebeurde, afhing van de leeftijd
van de personen in kwestie. Om te beginnen, konden juist jonge mannen en
vrouwen door onderwijs en erfenis vaardigheden en kapitaal krijgen waarmee
ze een nieuwe, betere baan konden verwerven. Dat geldt bijvoorbeeld voor boe-
renzonen die voor ze de boerderij van hun vader overnamen – of eventueel een
andere boerderij verworven – eerst een tijdje als landarbeider werkten. Er zullen
ook ongeschoolde arbeiders zijn geweest die zich schoolden om als ambtenaar,
onderwijzer of geschoolde fabrieksarbeider verder hun brood te verdienen. Ook
dat gebeurde eerder aan het begin dan aan het eind van een loopbaan. Daar-
naast is het zo dat jonge ongehuwde mannen en vrouwen doorgaans eerder van
baan en zelfs woonplaats wisselen dan mensen van middelbare en hogere leef-
tijd (Maas & Van Leeuwen, 2004). Die laatste groepen zitten meer vast aan hun
baan en plaats vanwege kinderen, vrienden en het feit dat ze al een tijd een baan
hebben. Dat laatste belemmerde des te meer een verhuizing als het moeilijk was
bij een bedrijf ‘binnen’ te komen (en dat was in de Nederlandse arbeidsmarkt
soms zo) of als een organisatie, zoals een bank, met een interne arbeidsmarkt
senioriteit beloonde met bijvoorbeeld een hoger loon, een ziekteverzekering of
een pensioen. We verwachten daarentegen dat de ouderdom vaker met neerwaartse mobi-
liteit verbonden was. Werkgevers konden proberen om oudere werknemers die
niet zo snel meer waren, of vaker ziek, te lozen of in ieder geval werk te geven
dat minder geschoold was en minder verdiende (Ransom & Sutch, 1986; Johnson,
1994; Bulder, 1993). Op grond van een analyse van de Nederlandse beroepentel-
lingen in de periode 1899-1947 is wel geconcludeerd dat de meeste bejaarde man-
nen bleven werken (Bulder, 1993). Dat geschiedde meer in beroepsgroepen waar-
in men zelf het arbeidstempo kon regelen zoals bij winkeliers en boeren, maar
weer niet in een fabriek. Het is mogelijk dat dit ook gold voor landarbeiders die
tegen een lager loon klusjes konden gaan doen, en voor ambachtsknechten. Men ineke maas en marco h.d. van leeuwen spreekt in zulke gevallen wel van ‘pensionering op de werkplek’ (retirement on the
job). Hypothesen over de ‘vorm’ van de beroepscarrière Hypothesen over veranderingen in de invloed van vaders beroepsklasse op het carrièrever
loop van zijn kinderen Hypothesen over veranderingen in de invloed van vaders beroepsklasse op het carrièrever
loop van zijn kinderen Een belangrijke veronderstelling over veranderingen van de invloed van de vader
op de beroepskansen van zijn kinderen, is dat deze afnam als gevolg van het ont-
staan van nieuwe, gespecialiseerde beroepen ten tijde van de industrialisering
(Treiman, 1970). Het ontbrak de ouderlijke generatie aan de gespecialiseerde ken-
nis, nodig voor deze beroepen, waardoor ze hun kinderen niet meer zelf konden
opleiden. De veranderingen in de beroepsstructuur en de toenemende speciali- van een dubbeltje naar een kwartje? sering leidden er ook toe dat het belang van onderwijs sterk toenam. Zoals uit de
bijdrage van Boonstra aan dit boek blijkt, waren er aan het eind van de periode
bijna geen mensen meer die hun handtekening niet konden zetten. Praktisch
iedereen leerde dus (lezen en) schrijven op de lagere school. En een steeds groter
deel van de bevolking volgde hoger onderwijs. Via de vaardigheden geleerd op
dat onderwijs en met via onderwijs verkregen diploma’s en kwalificaties konden
mensen vaker andere beroepen kiezen dan die van hun ouders. Omgekeerd werd
het voor werkgevers mogelijk, en voor sommige nieuwe gespecialiseerde beroe-
pen wellicht ook nodig, om te selecteren, niet op basis van het sociaal milieu van
de vaders, maar op basis van de diploma’s en kwalificaties van de betrokkenen
zelf. 180 Was er sprake van toenemende specialisatie in de onderzochte periode? Een
wijze om specialisatie op het spoor te komen is te kijken naar het aantal verschil-
lende beroepen die in een bepaalde periode worden uitgeoefend. De beroepsti-
tels zoals die in de eerdergenoemde Zeeuwse huwelijksakten genoteerd staan
zijn alle in HISCO gecodeerd (Van Leeuwen, Maas & Miles, 2002). HISCO telt onge-
veer 1600 rubrieken. Deze zijn zo gekozen dat zowel eeuwenoude taken in het
productieproces zoals bakker of historicus er een plek in hebben als ‘nieuwe’
taken in het productieproces zoals fabrieksarbeider of socioloog. Het aantal ver-
schillende beroepen van de bruidegoms nam toe van 236 in de periode 1856-1865
tot 337 in de periode 1916-1922 (zie figuur 2). Vooral na 1885 steeg het aantal
beroepen snel. Het aantal verschillende beroepen van de bruiden schommelde Figuur 2 Het aantal verschillende beroepen (HISCO-groepen) van Zeeuwse bruidegoms en
bruiden, naar periode
Bron: Van Leeuwen & Maas, 2007. Een voorbeeld: de loopbaan van Pieter Arkenbout
(1816 tot na 1890) Voordat we overgaan tot een toetsing van de hypothesen, laten we eerst aan de hand
van de autobiografie van Pieter Arkenbout een voorbeeld zien van een beroepscar-
rière die zich grotendeels afspeelde in de door ons onderzochte periode. 182 Zijn jeugd
11 December 1816 (…) zag ik te Zwartewaal het levenslicht (...) en was
mijn vader daar korenmolenaar. De sterke concurrentie en groote
vermeerdering van het huisgezin (wij zijn met ons negenen groot
geworden) waren oorzaak dat ik, en ook alle mijne broeders en zusters,
hoe lief onze ouders ons ook hadden, niet zoo lang school konden laten
gaan als zij wel gewenscht hadden, omdat dit in dien tijd veel geld koste
(...). En deed ik reeds als schooljongen mijn best om al wat mijne krachten
toelieten op den molen te leeren, en ging ik met 1 November 1828 van de
school om al spoedig de eenige hulp voor Vader te zijn. En nu? Ja dat was de vraag! eerst trachtte ik mij aan het burgerleven te
gewennen, en mij weer in het vak bij mijn vader te bekwamen, doch
begreep al spoedig dat ik hier de kost niet verdienen kon, daar het werk
vóór mijne komst toch ook gedaan werdt, waarom ik dan ook naar eene
dienst zocht. En die vond te Nieuwenhoorn bij den heer G.J. Haverkamp
bij wien ik mij verhuurde voor een jaar, ingaande Mei voor fl 72,– boven
kost, inwoning en bewassching. Nu weet ik wel dat menigeen die dit leest
zal zeggen “wat een beetje geld”, maar bedenk dat het toen nog maar 1840
was, en het nu ik dit schrijf reeds 1890 is. En dient ook nog dat hij mij fl
2,– meer gaf dan vorige knegts verdienden, omdat mijn vader een goede
kennis van hem was, en ook omdat mijn persoon hem goed aanstond. Na diverse baantjes, via advertenties of via via verkregen, wordt hij, inmiddels
gehuwd, in 1860 aangenomen als ‘Vader’ van het weeshuis te Brielle. En zeide ik het molenaarsvak waar ik in geboren en opgevoed was, vaar-
wel! Nu waren wij dan niet alleen in eene andere woonplaats, maar wat
meer zegt in een geheel andere betrekking in de maatschappij gekomen. Hoe wij in dien eersten tijd te moede waren laat zich niet zoo gemakkelijk
indenken, of men moet zelve eene dergelijke standverwisseling ondervon-
den hebben. Hypothesen over de ‘vorm’ van de beroepscarrière 0
50
100
150
200
250
300
350
400
1856-1865
1876-1885
1896-1905
1916-1922
periode
aantal
Bruidegom
Bruid Figuur 2 Het aantal verschillende beroepen (HISCO-groepen) van Zeeuwse bruidegoms en
bruiden, naar periode aantal Bruidegom
B
id Bruidegom Bruid 1916-1922 periode Bron: Van Leeuwen & Maas, 2007. Bron: Van Leeuwen & Maas, 2007. ineke maas en marco h.d. van leeuwen ineke maas en marco h.d. van leeuwen op een veel lager niveau iets onder de 40. Hier is pas na 1905 een kleine stijging
waarneembaar. Een andere methode is te kijken naar de verhouding tussen algemene en
gespecialiseerde beroepsgroepen (Van Leeuwen & Maas, 2007). Over de hele peri-
ode was er bij de mannen een toename van het aandeel gespecialiseerde beroe-
pen van 40 procent in de jaren 1856-1865 tot 70 procent in de jaren 1916-1922
(figuur 3). Ook bij de vrouwen steeg het percentage gespecialiseerde beroepsti-
tels. Na 1910 was er sprake van een kentering, vooral bij de vrouwen en in min-
dere mate bij de mannen. Het percentage gespecialiseerde beroepen lag lager bij
mannen dan bij vrouwen. Mannen stonden vaker geregistreerd als ‘arbeider’ of
‘werkman’, terwijl er bij vrouwen vaker werd genoteerd wat voor soort werk zij
deden, bijvoorbeeld dienstbode of boerenmeid. 181 Kortom, gedurende de onderzochte periode was er sprake van een toename
van nieuwe en gespecialiseerde beroepen. Op grond daarvan verwachten we dat
de invloed van de beroepsklasse van de vader op de beroepscarrière van zijn
kinderen afnam. Daarbij zijn twee kanttekeningen te maken. Ten eerste lijkt de
ontwikkeling sterker voor beroepen waarin mannen werken dan voor vrouwen-
beroepen. Ten tweede is het mogelijk dat de specialisering omstreeks 1910 een
(voorlopig) einde nam. Als gevolg daarvan zou ook de invloed van de vader ster-
ker afnemen voor zonen dan voor dochters en wellicht alleen tussen 1865 en
1910 en niet daarna. van een dubbeltje naar een kwartje
Figuur 3 Het aandeel van gespecialiseerde beroepsbeoefenaren onder Zeeuwse bruidegoms en
bruiden, naar periode
Bron: Van Leeuwen & Maas, 2007. 0
10
20
30
40
50
60
70
80
90
100
1856-1865
1876-1885
1896-1905
1916-1922
periode
%
Bruidegom
Bruid Figuur 3 Het aandeel van gespecialiseerde beroepsbeoefenaren onder Zeeuwse bruidegoms en
bruiden, naar periode uur 3 Het aandeel van gespecialiseerde beroepsbeoefenaren onder Zeeuwse bruidegoms en
bruiden, naar periode Bruidegom
Bruid 1916-1922 Bron: Van Leeuwen & Maas, 2007. van een dubbeltje naar een kwartje? van een dubbeltje naar een kwartje? Een voorbeeld: de loopbaan van Pieter Arkenbout
(1816 tot na 1890) Wij die van jongs af gewoon waren ondergeschikt te zijn en
anderen te dienen, zagen ons nu in eens aan het hoofd geplaatst [van het
weeshuis], dat behalve naaijuffrouw, dienstbode, wasch- en werkvrouwen,
22 kinderen bevatte. ineke maas en marco h.d. van leeuwen Na een conflict met de dominee ontslaan de bestuurders van het weeshuis hem in
1866. En zie! (...) met het Nieuwejaarsfeest kwam bij Mevrouw Verhey, die oud-
ste Regentesse was, en tegen ons ontslag gestemd had, een zoon over, die
Candidaat Notaris te Hillegersberg was en die, daar er over ons gespro-
ken werdt zeide, dat een ongehuwde docter aldaar met 1 february andere
huisbewaarders hebben moest. Wat zij ons liet weten, en wijl haar zoon
dit zoo mooi voorgesteld had, besloten wij dat ik daar eens zou gaan zien. En werdt het met dien heer al spoedig eens, want hij wist zich zoo goed
voor te doen, dat ik werkelijk geloofde dat dit een duurzame betrekking
worden zou, waarom wij er dan met 1 february naar toe gingen. Doch onze
goede verwachtingen werden al spoedig den bodem ingeslagen, want het
duurde niet lang of wij leerden hem wel beter kennen, want in plaats van
godsdienstig zoo als hij voorgegeven had, was hij een beslist ongeloovi-
ge, daarbij was hij zelfs meester in de vrijmetslaarsloge te Rotterdam. (…)
Waarom ik dan ook als met beide handen de gelegenheid aangreep die
mijn broeder Gerrit, die te Waddingsveen woonde, bij gelegenheid dat hij
ons kwam bezoeken, mij aan de hand deed om voor het [begrafenisverze-
kerings] fonds waarvan hij daar toen boekhouder was, de leden van een
bode over te nemen die er af wilde. (…) Zoo was ik dan nu in mijn derde
betrekking, nadat ik het molenaarsvak verlaten had, en hadden wij het,
zoo men wel denken kan, niet te ruim, maar al spoedig kreeg de vrouw
ook al wat naaiwerk, en had zij het soms druk, zij was nu weer haar eigen
meester, en zoo leefden wij gelukkig en tevreden. 183 Uiteindelijk vindt hij ander werk in 1868. j
j
Toen de zomer kwam ging ik op een dinsdagmiddag (…) naar de brug en
Zuidkade, om in dat deel der gemeente contributie te gaan ontvangen
toen ik den heer P. Verbruggen voor mij uit zag wandelen. (…) Ik had reeds
lang gewenscht die te kunnen spreken, omdat ik van mijn broeder G. Een voorbeeld: de loopbaan van Pieter Arkenbout
(1816 tot na 1890) en
ook van anderen, wel gehoord had, dat hij als timmerman en aannemer
van publieke werken, veel omhanden had, en nog al eens schrijfwerk door
anderen liet verrigten. En ik nu ook niet verzuimde hem mede te deelen,
dat ik veel tijd over had, met de vraag, of hij niet wat schrijfwerk voor mij
te doen had. Waarop hij mij antwoordde, dat hij dit nu wel reeds had, maar
iemand noodig had die met het vak bekend was, en dit dus mij niet kon
opdragen. Doch nadat wij nog een en ander omtrent het vak, waarmede ik
als weesvader en voornamelijk door ik in dien tijd veel voor mijn broeder
Jan die daar als timmerman dicht bij woonde, geschreven had, niet zoo
geheel vreemd er mee was, gesproken hadden, beloofde hij mij zoodra hij
eenig werk voor mij had om mij te zullen zenden. Waarmede hij dan ook
niet lang wachtte, want anderhalve week later, zond hij een boodschap
om bij hem te komen. En droeg hij mij toen op, om uit een bestek dat Toen de zomer kwam ging ik op een dinsdagmiddag (…) naar de brug en
Zuidkade, om in dat deel der gemeente contributie te gaan ontvangen
toen ik den heer P. Verbruggen voor mij uit zag wandelen. (…) Ik had reeds
lang gewenscht die te kunnen spreken, omdat ik van mijn broeder G. en
ook van anderen, wel gehoord had, dat hij als timmerman en aannemer
van publieke werken, veel omhanden had, en nog al eens schrijfwerk door
anderen liet verrigten. En ik nu ook niet verzuimde hem mede te deelen,
dat ik veel tijd over had, met de vraag, of hij niet wat schrijfwerk voor mij
te doen had. Waarop hij mij antwoordde, dat hij dit nu wel reeds had, maar
iemand noodig had die met het vak bekend was en dit dus mij niet kon van een dubbeltje naar een kwartje? hij mij gaf, alle houtwaren uit te trekken en daarvan geregelde staten te
maken, daar hij die levering op zijne beurt aan houtkoopers wilde uitbe-
steden, en (…) mij ook nog weer ander werk opdroeg. Een voorbeeld: de loopbaan van Pieter Arkenbout
(1816 tot na 1890) (...) Toen nu ook het
tweede mij opgedragen werk des zaterdagsmiddags thuisgebracht en ook
naar genoegen was, vroeg Verbruggen of ik elken zaterdag kon komen dan
kon hij mij een en ander omtrent den loop der zaak mededeelen en bij
het betalen der knegts tegenwoordig zijn, om, als hij niet thuis was, die te
kunnen betalen, en zijn vrouw er dan geen drukte mee had, en zoo ben ik
van lieverlede daar in de zaak gekomen, waar ik nu geregeld de 4 laatste
dagen der week doorbracht, waardoor mijne verdiensten, en wijl ook het
fonds reeds aardig was vooruitgegaan, zoo zijn toegenomen dat wij nu in
onzen stand meer fatsoenlijk konden rondkomen. (Geciteerd bij Van Leeuwen, 1992, 216-218) 184 De autobiografie van Pieter Arkenbout laat een grote mate van beroepsmobili-
teit zien, vooral in de tweede helft van zijn leven. Zijn familieleden, vader, zus en
broer, speelden een belangrijke rol. Op hun voorspraak en met hun informatie
weet hij van molenaarsknecht op te klimmen tot boekhouder. Een prestatie waar
hij ook zelf soms versteld van staat. De beroepscarrière De beroepscarrière Voor de analyse van de beroepscarrières maken we gebruik van alle beroepsge-
gevens van de onderzoekspersoon uit de registers en de gezinskaarten plus het
beroep in 1940 van de persoonsgezinskaart (voor zover bekend). Mensen werden
opgetekend in het bevolkingsregister bij veranderingen in hun gezinssamenstel-
ling (bijvoorbeeld de geboorte van een kind of sterfte van de partner) of bij ver-
huizingen. Bij die gelegenheid werd vaak het beroep genoteerd. Ook werd er na
elke volkstelling begonnen met een nieuwe registratie. Ook dit kon leiden tot het
vermelden van een beroep. Dus op bepaalde (niet direct aan de beroepscarrière
gerelateerde) tijdstippen werd het beroep vastgelegd. Het aantal beroepsvermel-
dingen verschilt daarbij tussen individuen. Wanneer iemand precies van beroep
veranderde is met deze gegevens niet vast te stellen. Men kan nog wel waarne-
men dat het beroep ergens tussen de geboorte van het eerste en de geboorte van
het tweede kind is veranderd, maar wanneer precies is onduidelijk. Omdat er voor het vijftiende en na het tachtigste levensjaar nauwelijks
beroepen zijn waargenomen, kijken we alleen naar mannen en vrouwen tussen
deze leeftijden. In totaal zijn er 4658 valide (codeerbare) beroepstitels met een
datering. Deze gegevens hebben betrekking op 2039 onderzoekspersonen waar-
van 1295 mannen en 744 vrouwen. De ondervertegenwoordiging van vrouwen
komt tot stand, doordat er meer vrouwen waren die zich nooit op de arbeids-
markt begaven, of dermate weinig frequent en kort dat ze niet waargenomen
werden op de arbeidsmarkt. Het grote verschil tussen het aantal personen met
beroepstitels en de omvang van het cohort, is verder voornamelijk een gevolg
van de hoge kindersterfte waardoor veel kinderen de vijftien jaar niet haalden. Gemiddeld worden er voor de onderzoekspersonen 2,3 beroepen waargeno-
men. Dit aantal verschilt niet tussen mannen en vrouwen. Bij mannen varieert
het aantal waarnemingen van beroepen tussen 1 en 10, bij vrouwen tussen 1 en
18. Informatiever is de ‘beroepsinformatieratio’, ofwel het aantal beroepswaarne-
mingen per persoon per jaar dat deze persoon kan worden waargenomen (Maas
& Van Leeuwen, 2004). De waarnemingsperiode begint als een persoon vijftien
wordt en eindigt bij overlijden of, voor incomplete levenslopen, op het moment
dat de dataverzameling stopt. De beroepsinformatieratio bedraagt zowel voor
mannen als voor vrouwen 0,06. Dat wil zeggen gemiddeld één beroepswaarne-
ming per 17 jaar. Figuur 4 toont deze ratio naar leeftijd. Verreweg de meeste
beroepen worden waargenomen op jonge leeftijd, rond het huwelijk. Steekproef Steekproef Voor een meer algemene beschrijving van de beroepscarrières van mannen en
vrouwen en voor een toetsing van de hypothesen maken we gebruik van de His-
torische Steekproef Nederlandse bevolking (HSN), dataset Levenslopen, release
2007.01. Deze data zijn verzameld in het project Life Courses in Context (Mande-
makers, 2004), zie ook de bijdrage van Mandemakers aan dit boek. Uitgangspunt
van de dataverzameling is een steekproef uit de geboorteaktes van de geboorte-
cohorten 1850-1922 voor de stad Rotterdam en de provincies Friesland, Zeeland
en Utrecht en de geboortecohorten 1883-1922 voor de rest van het land. Van al
deze personen zijn gegevens over de levensloop verzameld uit het bevolkingsre-
gister (achtereenvolgens de registers (in gebruik tot 1890), de gezinskaarten (tot
1940), de persoonsgezinskaarten (tot 1994) en de persoonslijsten). We analyseren hier alleen de geboortecohorten 1850-1882. Deze geboorte-
cohorten hebben hun actieve beroepsleven min of meer afgesloten in 1940 (de
jongste is dan 58). Dat is van belang, want in 1940 worden de persoonskaarten
ingevoerd. Op de persoonskaarten staan wel beroepen, maar deze zijn niet geda-
teerd. Alleen het eerste beroep op de persoonskaart (in het eerste huishouden)
is bruikbaar, omdat het hoogstwaarschijnlijk het beroep in 1940 was. Als gevolg
van deze selectie van cohorten beschikken we ook alleen over de provincies Fries-
land, Utrecht, Zeeland en de stad Rotterdam. Onze resultaten, vooral de beschrij-
vende, zijn daarom niet zomaar te generaliseren naar de rest van Nederland. ineke maas en marco h.d. van leeuwen De steekproef betrof 0,75 % van alle geboorteaktes voor de periode 1812-1872
en 0,50 % voor de periode 1873-1882. De verzameling van de levenslopen is nog
niet helemaal compleet in Friesland (ongeveer 75 % verzameld) en Rotterdam
(60 %). Ontbrekende personen zijn vooral degenen die heel veel of over een lange
afstand verhuisden. In totaal zijn er gegevens voor 5143 individuen. 185 De beroepscarrière Vooral bij
mannen stijgt het relatieve aantal waarnemingen weer op hogere leeftijd. Dit van een dubbeltje naar een kwartje? komt doordat voor meer mannen dan vrouwen in 1940 bij de overgang naar de
persoonsgezinskaarten een beroep is opgetekend. Alle beroepen zijn gecodeerd naar de Historical International Standard Clas-
sification of Occupations (HISCO) (Van Leeuwen, Maas & Miles, 2002) en daarna
is aan elke HISCO-categorie een beroepsstatus toegekend met behulp van de HIS-
CAM (v0.1) schaal (Maas e.a., 2006).2 De HISCO-categorieën zijn tevens omgezet
naar het klassenschema HISCLASS (Van Leeuwen & Maas, 2005). De 12 klassen
die in HISCLASS worden onderscheiden voegen we samen tot vier brede klassen:
witte boorden (HISCLASS 1, 2, 3, 4 en 5), geschoolde arbeiders (6 en 7), boeren (8)
en ongeschoolde arbeiders (9, 10, 11 en 12). 186 Andere variabelen De bevolkingsregisters geven ook informatie over het beroep van de vader, soms
zelfs meerdere keren. We selecteren het beroep van de vader dat zo kort mogelijk
voor of na de geboorte van de onderzoekspersoon is opgetekend. De beroepstitels
van de vaders zijn net als die van hun kinderen gecodeerd naar HISCO. Vervolgens
zijn ze omgezet naar HISCLASS en samengevoegd in dezelfde vier brede klassen
als de beroepen van de kinderen. Van alle onderzoekspersonen is de geboorte- Figuur 4 Beroepsinformatieratio bij mannen en vrouwen in de HSN-steekproef naar leeftijda
0,00
0,02
0,04
0,06
0,08
0,10
0,12
0,14
0,16
0,18
0,20
15
20
25
30
35
40
45
50
55
60
65
70
75
80
leeftijd
beroepsinformatieratio
Mannen
Vrouwen Figuur 4 Beroepsinformatieratio bij mannen en vrouwen in de HSN-steekproef naar leeftijda
a De beroepsinformatieratio geeft het aantal waarnemingen van beroepen per persoon per jaar. Uit-
gesplitst naar leeftijd is deze ratio bijna equivalent aan het percentage mannen en vrouwen met een
waarneming van beroep. Er is een klein verschil doordat sommige mannen en vrouwen meer dan een
beroepswaarneming per jaar hebben. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. 0,00
0,02
0,04
0,06
0,08
0,10
0,12
0,14
0,16
0,18
0,20
15
20
25
30
35
40
45
50
55
60
65
70
75
80
leeftijd
beroepsinformatieratio
Mannen
Vrouwen beroepsinformatieratio Mannen
Vrouwen Mannen Vrouwen a De beroepsinformatieratio geeft het aantal waarnemingen van beroepen per persoon per jaar. Uit-
gesplitst naar leeftijd is deze ratio bijna equivalent aan het percentage mannen en vrouwen met een
waarneming van beroep. Er is een klein verschil doordat sommige mannen en vrouwen meer dan een
beroepswaarneming per jaar hebben. a De beroepsinformatieratio geeft het aantal waarnemingen van beroepen per persoon per jaar. Uit-
gesplitst naar leeftijd is deze ratio bijna equivalent aan het percentage mannen en vrouwen met een
waarneming van beroep. Er is een klein verschil doordat sommige mannen en vrouwen meer dan een
beroepswaarneming per jaar hebben. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. ineke maas en marco h.d. van leeuwen datum bekend: het betreft hier immers een steekproef uit de geboorteakten. De
meting van een beroep is over het algemeen ook gedateerd, bijvoorbeeld omdat
het plaatsvond bij de geboorte van een kind.3 Met behulp van deze gegevens is de
leeftijd van mannen en vrouwen bij elke beroepsmeting vastgesteld. datum bekend: het betreft hier immers een steekproef uit de geboorteakten. Andere variabelen De
meting van een beroep is over het algemeen ook gedateerd, bijvoorbeeld omdat
het plaatsvond bij de geboorte van een kind.3 Met behulp van deze gegevens is de
leeftijd van mannen en vrouwen bij elke beroepsmeting vastgesteld. 187 Beschrijvende resultaten Mobiliteit Hoe zagen de beroepscarrières van mannen en vrouwen geboren tussen 1850 en
1882 eruit? Was de periode tussen 1865 en 1940, waarin zij op de arbeidsmarkt
waren, gekenmerkt door opwaartse mobiliteit? En zette die opwaartse mobiliteit
gedurende hun hele beroepscarrière door, of leidde de oude dag tot neerwaartse
mobiliteit? Om deze vragen te beantwoorden analyseren we eerst de veranderingen
in beroepsstatus van individuele mannen en vrouwen. We vergelijken steeds
twee achtereenvolgende beroepswaarnemingen van dezelfde persoon. Indien
het tweede beroep een hogere status heeft dan het eerste, dan is er sprake van
opwaartse mobiliteit; is het tweede beroep lager, dan van neerwaartse mobi-
liteit. Hebben beide beroepen dezelfde status, dan nemen we aan dat er geen
mobiliteit is geweest of alleen mobiliteit naar een beroep met precies dezelfde
status. Om enig zicht te krijgen op de timing van mobiliteit in de levensloop,
gaan we ervan uit dat de mobiliteit plaatsvond op een tijdstip midden tussen de
twee beroepswaarnemingen. De resultaten staan in tabel 1. De beroepscarrières van mannen laten veel meer mobiliteit zien dan die van
vrouwen. Als we twee achtereenvolgende beroepswaarnemingen vergelijken,
dan is bij vrouwen in slechts 17 procent van de gevallen sprake van mobiliteit, bij
mannen is dat in 41 procent van de gevallen. Zowel opwaartse mobiliteit kwam
bij mannen meer voor als neerwaartse mobiliteit. Echter, als er een verschil in
status was tussen twee opeenvolgende beroepen, dan was dat bij vrouwen gemid-
deld een groter verschil dan bij de mannen. Dit geldt zowel voor opwaartse mobi-
liteit (respectievelijk 31 statuspunten voor vrouwen en 15 voor mannen) als voor
neerwaartse mobiliteit (respectievelijk -25 en -9 statuspunten). We kijken vervolgens of mannen en vrouwen in verschillende beroepsklas-
sen zich onderscheiden in de kans om mobiel te zijn naar een beroep met een
hogere of een lagere status. Aangezien de beroepsklassen beroepen bevatten met
verschillende status, kan dit dus zowel mobiliteit binnen een beroepsklasse zijn
als mobiliteit waarbij een klassengrens wordt overschreden. Bij mannen is, zoals
verwacht, de kans op mobiliteit het grootst voor de ongeschoolde arbeiders (47,5
%). Dit geldt vooral voor opwaartse mobiliteit. Neerwaartse mobiliteit kwam het
meest voor onder de boeren. Bij vrouwen zien we geen verschillen tussen de onge-
schoolde arbeiders en de witteboordenklasse wat betreft totale mobiliteit. Bij
nader inzien is dit een gevolg van de grotere kans op opwaartse mobiliteit voor van een dubbeltje naar een kwartje? Beschrijvende resultaten Tabel 1 Mobiliteit van mannen en vrouwen in Nederland tussen 1865 en 1940, naar leeftijd,
sociale klasse en periode (percentages en gemiddeld verschil in status) a Tabel 1 Mobiliteit van mannen en vrouwen in Nederland tussen 1865 en 1940, naar leeftijd,
sociale klasse en periode (percentages en gemiddeld verschil in status) a sociale klasse en periode (percentages en gemiddeld verschil in status)
Totaal
(%)
Op-
waarts
(%)
Neer-
waarts
(%)
<1900
(%)
>1900
(%)
Op-
waartse
afstand
(M)
Neer-
waartse
afstand
(M)
Mobiliteit mannen
Totaal
40,8
23,3
17,5
15,1
-9,1
Sociale klasseb
Witte boorden
33,8*
14,2*
19,6*
Geschoolde arbeiders
26,2
15,1
11,1
Boeren
34,3
7,8
26,5
Ongeschoolde arbeiders 47,5
29,6
17,8
Leeftijd
15-29
36,6*
19,5*
17,0
35,4
42,6
16,9
-9,3
30-49
50,4
30,4
20,0
41,4
53,6^
13,4
-8,5
50-80
34,7
19,7
15,0
- c
34,7
-
-
Mobiliteit vrouwen
Totaal
17,1
12,4
4,7
31,2
-25,1
Sociale klasseb
Witte boorden
13,9
3,0*
10,9*
Geschoolde arbeiders
-
-
-
Boeren
-
-
-
Ongeschoolde arbeiders
17,2
13,7
3,5
Leeftijd
15-29
12,8*
9,0*
3,8*
12,4
16,7
32,8
-
30-49
36,7
28,9
7,8
29,9
41,4
29,7
-
50-80
13,3
4,4
8,9
-
-
-
-
* = p < 0,05, toets op verschillen tussen alle categorieën;
^ = p < 0,05, toets op verschillen tussen de periodes binnen een leeftijdscategorie. a Vergelijking van steeds twee opeenvolgende beroepswaarnemingen. De leeftijd waarop mobiliteit plaat-
vond is niet precies bekend, maar geschat als het midden tussen de twee beroepswaarnemingen. b Beroepsklasse van de eerste beroepswaarneming. c - = N kleiner dan 50. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-1882. 188 * = p < 0,05, toets op verschillen tussen alle categorieën; Beroepsklasse van de eerste beroepswaarneming. p
p
g
- = N kleiner dan 50. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-1882. ineke maas en marco h.d. van leeuwen ineke maas en marco h.d. van leeuwen vrouwen uit de eerstgenoemde klasse en een grotere kans op neerwaartse mobi-
liteit voor vrouwen uit de tweede klasse. Er zijn te weinig vrouwelijke geschoolde
arbeiders en boerinnen in de data om daar een uitspraak over te doen. 189 Opwaartse mobiliteit kwam vaker voor dan neerwaartse mobiliteit, bij man-
nen 1,5 keer zo vaak, bij vrouwen 2,5 keer. We zien dit ook in bijna alle leeftijds-
groepen. De enige uitzondering hierop vormen de vrouwen boven de vijftig. Hun
kans op opwaartse mobiliteit was slechts 4,4 procent tegen 8,9 voor neerwaartse
mobiliteit. De kans op opwaartse mobiliteit was het grootst op middelbare leeftijd. Neerwaartse mobiliteit hing bij mannen niet significant samen met leeftijd. Bij
vrouwen nam de kans op neerwaartse mobiliteit toe naarmate zij ouder werden. Onze verwachtingen over de vorm van de beroepscarrière worden dus nauwelijks
bevestigd. Er is relatief gezien niet meer opwaartse mobiliteit op jonge leeftijd
dan later en de kans op neerwaartse mobiliteit neemt alleen bij vrouwen licht
toe over de levensloop. Een vergelijking tussen periodes is alleen mogelijk voor specifieke leeftijds-
groepen. De data bevatten immers geen 50-plussers voor 1900 en geen heel jonge
mensen na 1900. Hoewel alleen significant voor mannen tussen de 30 en 49,
wijzen de gegevens voor alle leeftijdsgroepen op een toename van de mobiliteit. Bij de mannen nam de kans op mobiliteit voor de 30-49-jarigen bijvoorbeeld toe
van 41 naar 54 procent, bij vrouwen van deze leeftijd was er een toename van 30
naar 41 procent. Gemiddelde beroepsstatus 10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
55
60
65
70
75
80
leeftijd
gemiddelde beroepsstatus
waargenomen
volgens model
10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
55
60
65
70
78
leeftijd
gemiddelde beroepsstatus
waargenomen
volgens model Figuur 5 Gemiddelde beroepsstatus van mannen en vrouwen in Nederland tussen 1865 en
1940, naar leeftijd
M Figuur 5 Gemiddelde beroepsstatus van mannen en vrouwen in Nederland tussen 1865 en
1940, naar leeftijd
Mannen
10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
55
60
65
70
75
80
leeftijd
gemiddelde beroepsstatus
waargenomen
volgens model 1940, naar leeftijd
Mannen
10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
55
60
65
70
75
80
leeftijd
gemiddelde beroepsstatus
waargenomen
volgens model 190
ineke maas en marco h.d. van leeuwen
1940, naar leeftijd
Mannen
N = 1295 (2956 beroepen), model: status = 44,6 + 0,37 * leeftijd – 0,005 * l
begint bij 15. Vrouwen
N = 744 (1702 beroepen), model: status = 12,1 + 1,36 * leeftijd – 0,12 * leeftijd
bij 15. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. 10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
55
60
65
70
75
leeftijd
gemiddelde beroepsstatus
10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
55
60
65
70
78
leeftijd
gemiddelde beroepsstatus 190 waargenomen
volgens model 1295 (2956 beroepen), model: status = 44,6 + 0,37 * leeftijd – 0,005 * leeftijd2, waarbij leeftijd
nt bij 15. Vrouwen
N = 744 (1702 beroepen), model: status = 12,1 + 1,36 * leeftijd – 0,12 * leeftijd2, waarbij leeftijd begint
bij 15. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. 10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
55
60
65
70
78
leeftijd
gemiddelde beroepsstatus
waargenomen
volgens model Vrouwen Vrouwen
10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
55
60
65
70
78
leeftijd
gemiddelde beroepsstatus
waargenomen
volgens model waargenomen
volgens model N = 744 (1702 beroepen), model: status = 12,1 + 1,36 * leeftijd – 0,12 * leeftijd2, waarbij leeftijd begint
bij 15. Gemiddelde beroepsstatus Een tweede antwoord op onze vragen krijgen we door te kijken naar de gemid-
delde beroepsstatus van mannen en vrouwen naar leeftijd (figuur 5). De gemid-
delde beroepsstatus van mannen veranderde nauwelijks over hun levensloop. Voor hun 25ste levensjaar was de gemiddelde beroepsstatus ongeveer 45 pun-
ten op de gebruikte statusschaal, daarna varieerde ze rond de 48 punten. Als de
beroepsstatus wordt gemodelleerd als een kwadratische functie van leeftijd, dan
lijkt er sprake te zijn geweest van een afnemende stijging van de gemiddelde
beroepsstatus naarmate mannen ouder werden. Tot een daling van de gemid-
delde beroepsstatus komt het echter niet of nauwelijks. De gemiddelde beroepsstatus van vrouwen naar leeftijd oogt heel anders. Oudere vrouwen hadden gemiddeld een 30 punten hogere beroepsstatus dan
jonge vrouwen. Het verschil met de mannen openbaart zich vooral op jonge leef-
tijd. Vrouwen waren op die leeftijd werkzaam in beroepen met een heel lage
status, tussen de 10 en 20, terwijl mannen op die leeftijd gemiddeld al een status
hadden bereikt die vrouwen pas aan het eind van hun carrière bereikten. De
gemiddelde beroepsstatus van vrouwen nam vooral toe tussen hun 15de en 30ste
levensjaar. Daarna vlakt de stijging af. Een belangrijk nadeel van figuur 5 is dat er geen onderscheid wordt gemaakt van een dubbeltje naar een kwartje? Figuur 5 Gemiddelde beroepsstatus van mannen en vrouwen in Nederland tussen 1865 en
1940, naar leeftijd
Mannen
N = 1295 (2956 beroepen), model: status = 44,6 + 0,37 * leeftijd – 0,005 * leeftijd2, waarbij leeftijd
begint bij 15. Vrouwen
N = 744 (1702 beroepen), model: status = 12,1 + 1,36 * leeftijd – 0,12 * leeftijd2, waarbij leeftijd begint
bij 15. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. Gemiddelde beroepsstatus Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. ineke maas en marco h.d. van leeuwen ineke maas en marco h.d. van leeuwen tussen de verschillen in beroepsstatus tussen individuen en de veranderingen
‘binnen’ individuen, dat wil zeggen in de loop van het leven van een en dezelfde
man of vrouw. Alleen die laatste hebben betrekking op beroepsmobiliteit. De
sterke stijging van de gemiddelde beroepsstatus van vrouwen zou bijvoorbeeld
het resultaat kunnen zijn van het selectief betreden en verlaten van de arbeids-
markt. Een eerste groep van vrouwen zou de arbeidsmarkt op jonge leeftijd kun-
nen betreden, in beroepen met een lage status werken en ook op relatief jonge
leeftijd de arbeidsmarkt weer kunnen verlaten. Een tweede groep zou kunnen
bestaan uit vrouwen die de arbeidsmarkt pas laat betreden en die dan werken in
beroepen met een relatief hoge status. De gemiddelde status naar leeftijd zou in
dat geval hetzelfde verloop kennen als in figuur 5. 191 Om beter zicht te krijgen op dit mogelijke selectieve betreden en verlaten
van de arbeidsmarkt maken we aparte figuren voor mannen en vrouwen die al
op jonge leeftijd op de arbeidsmarkt waren en mannen en vrouwen die op hoge
leeftijd nog op de arbeidsmarkt waren (figuur 6a en 6b). Zoals te verwachten
viel, zijn er bij de vrouwen meer aanwijzingen voor selectiviteit dan bij de man-
nen. Het verloop van de gemiddelde beroepsstatus naar leeftijd lijkt bij man-
nen die vroeg op de arbeidsmarkt kwamen veel op het verloop bij mannen die
nog op oudere leeftijd op de arbeidsmarkt waren (N.B. voor een klein deel, in 34
gevallen, zijn dit dezelfde mannen). Bij vrouwen zien we meer verschil. Vrouwen
die op latere leeftijd – na hun 50ste – nog, of weer, op de arbeidsmarkt waren,
lijken vlakkere beroepscarrières te hebben gehad dan vrouwen die al jong op de
arbeidsmarkt waren. De eerste groep begon gemiddeld vaker al op een hoger
niveau en hield dat niveau vast tot op hoge leeftijd. De carrières van vrouwen die
voor hun 20ste al op de arbeidsmarkt waren, laten gemiddeld een monotonere
stijging zien. Dus als deze vrouwen op de arbeidsmarkt bleven, dan verbeterden
zij over het algemeen hun positie. Negentien vrouwen waren zowel voor hun
20ste op de arbeidsmarkt als na hun 50ste. Gemiddelde beroepsstatus Kortom, de beroepscarrières van mannen en vrouwen in de onderzochte
periode werden gekenmerkt door een aanzienlijke mate van stabiliteit, tenmin-
ste als het gaat om de status van de beroepen. Opwaartse mobiliteit kwam wel
meer voor dan neerwaartse. Vrouwen die op hogere leeftijd nog op de arbeids-
markt zijn hebben een gemiddeld hogere status dan de jonge vrouwen op de
arbeidsmarkt. Dat is echter deels een gevolg van selectieve in- en uittreding van
vrouwen met hoge en lage status. Toetsing van de hypothesen We schatten thans enkele multiniveaumodellen om onze hypothesen te toetsen. Op deze manier wordt er rekening mee gehouden dat er meerdere metingen
van beroepsstatus per persoon zijn (Hox, 2002). De resultaten voor de mannen
staan in tabel 2, die voor vrouwen in tabel 3. In het eerste, lege model wordt
slechts geschat hoeveel van de variantie in beroepsstatus tussen individuen is van een dubbeltje naar een kwartje? Figuur 6a Gemiddelde beroepsstatus van mannen in Nederland tussen 1865 en 1940, naar
leeftijd en uitgesplitst naar ‘jonge’ en ‘oude’ werkers Figuur 6a Gemiddelde beroepsstatus van mannen in Nederland tussen 1865 en 1940, naar
leeftijd en uitgesplitst naar ‘jonge’ en ‘oude’ werkers 192
neke maas en marco h.d. van leeuwen
leeftijd en uitgesplitst naar jonge en oude werkers
Jonge werkers: mannen die voor hun 20ste al de arbeidsmarkt betraden
N = 365 (1130 beroepen ), model: status = 44,1 + 0,40 * leeftijd – 0,006 * leeftijd2, waarbij leeftijd begin
bij 15. Oude werkers: mannen die zich na hun 65ste nog op de arbeidsmarkt bevonden
N = 176 (543 beroepen), model: status = 38,1 + 0,59 * leeftijd – 0,006 * leeftijd2, waarbij leeftijd begin
bij 15. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. 10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
56
61
66
71
76
leeftijd
gemiddelde beroepsstatus
waargenomen
volgens model
10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
55
60
65
70
78
leeftijd
gemiddelde beroepsstatus
waargenomen
volgens mode Jonge werkers: mannen die voor hun 20ste al de arbeidsmarkt betraden
10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
56
61
66
71
76
leeftijd
gemiddelde beroepsstatus
waargenomen
volgens model Jonge werkers: mannen die voor hun 20ste al de arbeidsmarkt betraden
N = 365 (1130 beroepen ), model: status = 44,1 + 0,40 * leeftijd – 0,006 * leeftijd2, waarbij leeftijd begint
bij 15. Toetsing van de hypothesen 10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
56
61
66
71
76
leeftijd
gemiddelde beroepsstatus
waargenomen
volgens model Jonge werkers: mannen die voor hun 20ste al de arbeidsmarkt betraden Jonge werkers: mannen die voor hun 20ste al de arbeidsmarkt betraden N = 365 (1130 beroepen ), model: status = 44,1 + 0,40 * leeftijd – 0,006 * leeftijd2, waarbij leeftijd begint
bij 15. N = 365 (1130 beroepen ), model: status = 44,1 + 0,40 * leeftijd – 0,006 * leeftijd2, waarbij leeftijd begint
bij 15. N = 365 (1130 beroepen ), model: status = 44,1 + 0,40 * leeftijd – 0,006 * leeftijd2, waarbij leeftijd begint
bij 15. Oude werkers: mannen die zich na hun 65ste nog op de arbeidsmarkt bevonden Oude werkers: mannen die zich na hun 65ste nog op de arbeidsmarkt bevonden
N = 176 (543 beroepen), model: status = 38,1 + 0,59 * leeftijd – 0,006 * leeftijd2, waarbij leeftijd begint
bij 15. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. 10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
55
60
65
70
78
leeftijd
gemiddelde beroepsstatus
waargenomen
volgens model waargenomen
volgens model N = 176 (543 beroepen), model: status = 38,1 + 0,59 * leeftijd – 0,006 * leeftijd2, waarbij leeftijd begint
bij 15. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. j
Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. ineke maas en marco h.d. van leeuwen Figuur 6b Gemiddelde beroepsstatus van vrouwen in Nederland tussen 1865 en 1940, naar
leeftijd en uitgesplitst naar ‘jonge’ en ‘oude’ werkers Jonge werkers: vrouwen die voor hun 20ste al de arbeidsmarkt betraden
N = 395 (1026 beroepen), model: status = 13,8 + 0,54 * leeftijd + 0,04 * leeftijd2, waarbij leeftijd
begint bij 15. 10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
53
61
67
80
leeftijd
gemiddelde beroepsstatus
waargenomen
volgens model Jonge werkers: vrouwen die voor hun 20ste al de arbeidsmarkt betraden
10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
53
61
67
80
leeftijd
gemiddelde beroepsstatus
waargenomen
volgens model onge werkers: vrouwen die voor hun 20ste al de arbeidsmarkt betraden 193 N = 395 (1026 beroepen), model: status = 13,8 + 0,54 * leeftijd + 0,04 * leeftijd2, waarbij leeftijd
begint bij 15. Toetsing van de hypothesen Oude werkers: vrouwen die zich na hun 50ste nog op de arbeidsmarkt bevonden Oude werkers: vrouwen die zich na hun 50ste nog op de arbeidsmarkt bevonden
N = 80 (254 beroepen), model: status = 14,1 + 1,51 * leeftijd – 0,016 * leeftijd2, waarbij leeftijd begint
bij 15. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. 10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
55
60
65
70
78
leeftijd
gemiddelde beroepsstatus
waargenomen
volgens model 10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
55
60
65
70
78
leeftijd
gemiddelde beroepsstatus
waargenomen
volgens model N = 80 (254 beroepen), model: status = 14,1 + 1,51 * leeftijd – 0,016 * leeftijd2, waarbij leeftijd begint
bij 15. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. N = 80 (254 beroepen), model: status = 14,1 + 1,51 * leeftijd – 0,016 * leeftijd2, waarbij leeftijd begint
bij 15. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. j
Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. van een dubbeltje naar een kwartje? van een dubbeltje naar een kwartje? en hoeveel binnen de beroepscarrières van individuen. Voor zowel mannen als
vrouwen geldt dat er meer variantie is tussen individuen dan binnen carrières. Van de totale variantie in beroepsstatus bij de mannen ligt 73 procent tussen
individuen, bij de vrouwen 70 procent. 194 In het tweede model schatten we parameters voor de vorm van de beroeps-
carrière (lineaire en kwadratische effecten van leeftijd), van de historische tijd
en van de beroepsklasse van de vader. In overeenkomst met wat we al gezien heb-
ben in figuur 5 zien we ook hier dat de beroepsstatus van vrouwen aanzienlijk
sneller toeneemt met de leeftijd dan bij mannen. Bij mannen is bovendien deze
toename steeds geringer naarmate zij ouder worden. Na 21 jaar op de arbeids-
markt (op 36-jarige leeftijd) is de stijging van de gemiddelde beroepsstatus ten
einde en begint de daling. Bij vrouwen zien we zo’n kromming niet. Toetsing van de hypothesen Zoals eerder
al getoond is een deel van de stijging van de beroepsstatus van de vrouwen echter el 2 Multiniveau-analyse van de beroepsstatus van mannen in Nederland tussen 1865 en
1940 (coëfficiënten en significantieniveaus) Tabel 2 Multiniveau-analyse van de beroepsstatus van mannen in Nederland tussen 1865 en
1940 (coëfficiënten en significantieniveaus) Model 1
Model 2
Model 3
Model 4
Intercept
37,32 **
36,94 **
36,81 **
Leeftijd/10
3,82 **
4,20 **
4,18 **
Leeftijd/102
-0,93 **
-0,93 **
-0,93 **
Jaar (sinds 1865)
0,12 **
0,12 **
0,13 **
Vaders klassea
Witte boorden
14,38 **
14,82 **
14,89 **
Geschoolde arb. 10,05 **
12,46 **
12,20 **
Boeren
2,30 *
1,88
3,16
Leeftijd x vaders klasse
Witte boorden
-0,40
Geschoolde arb. -2,19 **
-2,41
Boeren
0,24
Jaar (sinds 1865) x
vaders klasse
Witte boorden
-0,02
Geschoolde arb. 0,19
Boeren
-0,32
Variantie tussen individuen
164,4
129,3
130,2
130,3
Variantie binnen individuen
60,6
58,0
57,3
57,3
a Referentiecategorie: ongeschoolde arbeiders. * = p < 0,05; ** = p < 0,01. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-1882. a Referentiecategorie: ongeschoolde arbeiders. * = p < 0,05; ** = p < 0,01. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-1882. ineke maas en marco h.d. van leeuwen vermoedelijk een gevolg van selectieve toe- en uittreding van de arbeidsmarkt. Er is een algemene opwaartse trend in de gemiddelde beroepsstatus van
zowel mannen als vrouwen. In 1940 ligt de gemiddelde beroepsstatus bij de man-
nen 9 punten hoger dan in 1865, bij de vrouwen zijn dat zelfs 13 punten. Er
zijn daarnaast duidelijke verschillen in beroepsstatus tussen mannen en vrou-
wen afkomstig uit verschillende herkomstklassen. Mannen en vrouwen van wie
de vader een witteboordenberoep uitoefende hebben beroepen die gemiddeld
respectievelijk 14 en 11 punten hoger scoren op de statusladder dan mannen
en vrouwen waarvan de vader een ongeschoolde arbeider was. Daarbij is er ove-
rigens sprake van een aanzienlijke ‘regressie naar het gemiddelde’. Bij de vaders
zelf was namelijk het verschil in status tussen de witteboordenklasse en de onge-
schoolde arbeiders maar liefst 23 punten. Ook de kinderen van geschoolde arbei-
ders scoren aanzienlijk hoger dan die van ongeschoolde arbeiders. Toetsing van de hypothesen De boerenkin- 195 bel 3 Multiniveau-analyse van de beroepsstatus van vrouwen in Nederland tussen 1865 en
1940 (coëfficiënten en significantieniveaus) Tabel 3 Multiniveau-analyse van de beroepsstatus van vrouwen in Nederland tussen 1865 en
1940 (coëfficiënten en significantieniveaus) Model 1
Model 2
Model 3
Model 4
Intercept
10,14 **
9,97 **
10,22 **
Leeftijd/10
6,93 **
7,04 **
6,72 **
Leeftijd/102
-0,30
-0,19
-0,19
Jaar (sinds 1865)
0,17 *
0,17 *
0,17 *
Vaders klassea
Witte boorden
10,62 **
9,54 **
6,00
Geschoolde arb. 6,29 **
8,06 **
10,19 **
Boeren
4,29
6,90 *
7,48
Leeftijd x vaders klasse
Witte boorden
1,00
Geschoolde arb. -1,80
Boeren
-3,00
Jaar (sinds 1865) x
vaders klasse
Witte boorden
0,17
Geschoolde arb. -0,14
Boeren
-0,13
Variantie tussen individuen
280,1
214,1
214,5
213,6
Variantie binnen individuen
118,9
92,8
92,4
92,5
a Referentiecategorie: ongeschoolde arbeiders. *
0 05 **
0 01 a Referentiecategorie: ongeschoolde arbeiders. * = p < 0,05; ** = p < 0,01. * = p < 0,05; ** = p < 0,01. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-1882. Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-1882. van een dubbeltje naar een kwartje? van een dubbeltje naar een kwartje? deren onderscheiden zich daarentegen niet veel in status van de kinderen van
ongeschoolde arbeiders. Vermoedelijk verlieten velen van hen het boerenbedrijf
om ongeschoolde arbeider te worden. 196 In het derde model kijken we of de beroepsklasse van de vader niet alleen Figuur 7 Gemodelleerde samenhang tussen beroepsstatus, vaders klasse en leeftijd van man-
nen en vrouwen in Nederland, beste model, 1900 Figuur 7 Gemodelleerde samenhang tussen beroepsstatus, vaders klasse en leeftijd van man-
nen en vrouwen in Nederland, beste model, 1900 mannen
vrouwen
Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. Toetsing van de hypothesen 10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
55
leeftijd
voorspelde beroepsstatus
vader witte-
boordenberoep
vader geschoolde
arbeider
vader boer
vader
ongeschoolde
arbeider
10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
55
leeftijd
voorspelde beroepsstatus
vader witte-
boordenberoep
vader geschoolde
arbeider
vader boer
vader
ongeschoolde
arbeider mannen
10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
55
leeftijd
voorspelde beroepsstatus
vader witte-
boordenberoep
vader geschoolde
arbeider
vader boer
vader
ongeschoolde
arbeider voorspelde beroepsstatus vader
ongeschoolde
arbeider vrouwen vrouwen
Bron: HSN dataset Levenslopen release 2007 01 geboortecohorten 1850 82
10
20
30
40
50
60
70
80
90
15
20
25
30
35
40
45
50
55
leeftijd
voorspelde beroepsstatus
vader witte-
boordenberoep
vader geschoolde
arbeider
vader boer
vader
ongeschoolde
arbeider vader
ongeschoolde
arbeider Bron: HSN-dataset Levenslopen, release 2007.01, geboortecohorten 1850-82. ineke maas en marco h.d. van leeuwen ineke maas en marco h.d. van leeuwen een effect had op de aanvangsstatus van de beroepscarrière van het kind, maar
ook op toename van status tijdens de levensloop. Dit blijkt slechts in geringe
mate het geval te zijn. Alleen bij de mannen vinden we dat geschoolde arbei-
ders minder snel stijgen dan zonen uit de overige klassen. De kromming van
de beroepscarrières blijkt niet te verschillen tussen de klassen (niet getoond in
de tabel). Het resultaat van deze twee bevindingen is dat zonen van geschoolde
arbeiders weliswaar op een behoorlijk hoog niveau hun beroepscarrière begin-
nen, maar daarna eigenlijk alleen maar status verliezen. De carrières van vrou-
wen verlopen daarentegen in gelijke mate opwaarts voor alle herkomstklassen
(vergelijk figuur 7). 197 Tot slot zien we in model 4 dat het effect van vaders beroepsklasse niet merk-
baar toe- of afgenomen is in de onderzochte periode. We toetsten ook of het
uitzonderlijke verloop van de beroepscarrières van de zonen uit de geschoolde
arbeidersklasse wellicht verdwijnt met de tijd, maar dat bleek niet het geval. Al
met al vinden we bijzonder weinig aanwijzingen voor historische veranderingen
in de beroepscarrières van mannen en vrouwen. Het enige is een langzame stij-
ging van de gemiddelde beroepsstatus voor alle groepen en op alle leeftijden. Conclusie Onze eerste, beschrijvende, vragen luidden: in welke mate kunnen de beroeps-
loopbanen van Nederlandse mannen en vrouwen geboren tussen 1865 en 1940
gekenmerkt worden als stabiel, opwaarts mobiel of neerwaarts mobiel? En: welke
veranderingen treden er op in de loop van de tijd? We gebruikten verschillende
methoden om deze vragen te beantwoorden. Dat bleek een bruikbare strategie. Zo liet een vergelijking van de beroepsstatus van steeds twee opeenvolgende
beroepen een hoge mate van mobiliteit zien bij de mannen en een aanzienlijk
geringere mobiliteit bij de vrouwen. Beide groepen hadden een aanzienlijk gro-
tere kans op opwaartse dan op neerwaartse mobiliteit. Als we kijken naar de
gemiddelde beroepsstatus van mannen en vrouwen over hun levensloop, dan
zien we echter dat de grote hoeveelheid mobiliteit bij mannen niet resulteerde
in grote veranderingen in de gemiddelde beroepsstatus. Er was dus wel sprake
van veel beweging op de arbeidsmarkt, maar niet van systematische stijging of
daling in de loop van een leven. Daarentegen hadden oudere vrouwen gemiddeld
wel een aanzienlijk hogere beroepsstatus dan jonge vrouwen. Deze verschillen
zijn te verklaren door de gemiddeld grotere afstand waarover vrouwen mobiel
waren – in dat geval kan men van een opwaartse carrière spreken –, maar moge-
lijk ook door selectieve in- en uittreding van de arbeidsmarkt door vrouwen, dat
wil zeggen dat de gegevens over de vroege loopbaan op andere vrouwen in een
andere arbeidsmarktsituatie betrekking hebben dan die over de late loopbaan. Hoe dan ook, in de eerste helft van de twintigste eeuw was er meer beroeps-
mobiliteit dan in de tweede helft van de negentiende eeuw, zowel voor vrou-
wen als voor mannen. Ook blijkt er sprake te zijn geweest van een toename van van een dubbeltje naar een kwartje? de gemiddelde beroepsstatus in de loop van de tijd (naast de verandering van
beroepsstatus over de levensloop). Deze toename verwachtten we ook gezien de
algemene afname van het aandeel beroepen met een lage status op de arbeids-
markt. We hadden ook verwacht dat deze ‘opwaardering’ van de arbeidsmarkt
samen met de afnemende seizoensarbeid zou leiden tot een vermindering van
de hoeveelheid mobiliteit. Dit blijkt echter niet het geval te zijn. Er is meer bewe-
ging in de beroepscarrières na 1900 dan ervoor, vooral bij mannen. Conclusie 198 Onze tweede reeks van vragen had betrekking op de mate waarin, en de
wijze waarop, beroepsloopbanen werden beïnvloed door de beroepsklasse van
de vader, met inbegrip van de vraag of deze invloed in de loop van de tijd ver-
anderde. Onze analyses laten een sterke samenhang zien tussen beroepsstatus
van de vader en de beroepsstatus van zijn kinderen. Deze samenhang komt met
name tot stand doordat zonen en dochters uit de hogere klassen hun beroeps-
carrière op een hoger niveau begonnen. We vonden nauwelijks verschillen in de
snelheid waarmee mannen en vrouwen carrière maakten tussen de herkomst-
klassen. Een uitzondering hierop vormen zonen van geschoolde arbeiders. Zij
begonnen hun carrière in beroepen met een relatief hoge status, maar bleven
daarna achter bij zonen afkomstig uit andere sociale klassen. Dochters bleken
over het algemeen minder te profiteren van hun herkomstklasse dan zonen. De
beginnende beroepsspecialisatie en de industrialisering in Nederland hebben,
tegen de verwachting in, niet geleid tot een afname van de invloed van het gezin
van herkomst op de beroepscarrière van hun kinderen. Behalve antwoorden op onze vragen levert deze studie nog enkele algeme-
ne conclusies op. De data verzameld in het kader van de Historische Steekproef
Nederland blijken genoeg informatie te bevatten over de beroepen van mannen
en vrouwen op verschillende tijdstippen in hun leven om interessante uitspraken
te doen over hun beroepscarrières. Daarbij lijkt het aan te bevelen om verschil-
lende methoden te gebruiken bij de analyse van de data. Zo leidden de analyses
van mobiliteit tot een ander beeld van de beroepscarrières dan de analyses van
gemiddelde beroepsstatus over de levensloop. Tot slot heeft deze studie nieuwe
vragen opgeworpen. Welke rol speelt bijvoorbeeld het selectieve in- en uittreden
van de arbeidsmarkt in de verklaring van de relatief hoge gemiddelde beroeps-
status van oudere vrouwen? Als de herkomstklasse nauwelijks van invloed is
op het al dan niet maken van carrière, en de snelheid daarvan, hoe kunnen de
gevonden verschillen in succes op de arbeidsmarkt dan wel worden verklaard? 1. Wij bedanken de deelnemers van het Utrechtse Migration and Social Stratification semi-
nar, in het bijzonder Frank van Tubergen, voor hun constructieve commentaar op eerdere
versies van deze tekst. Noten 1. Wij bedanken de deelnemers van het Utrechtse Migration and Social Stratification semi-
nar, in het bijzonder Frank van Tubergen, voor hun constructieve commentaar op eerdere
versies van deze tekst. 2. HISCAM is een historische versie van de CAMSIS-schalen. Deze schalen zijn gebaseerd op
de gedachte dat een ruime mate van interactie tussen personen met twee verschillende
beroepen betekent dat deze beroepen wat betreft status niet veel van elkaar verschillen 2. HISCAM is een historische versie van de CAMSIS-schalen. Deze schalen zijn gebaseerd op
de gedachte dat een ruime mate van interactie tussen personen met twee verschillende
beroepen betekent dat deze beroepen wat betreft status niet veel van elkaar verschillen ineke maas en marco h.d. van leeuwen (Prandy, 2000). Voor de schatting van de HISCAM-schaal zijn data gebruikt van 1,5 mil-
joen huwelijksaktes uit de periode 1800-1938 uit zes landen (Engeland, Canada, Frankrijk,
Duitsland, Zweden en Nederland). Als huwelijken tussen mannen en vrouwen met twee
verschillende beroepen relatief vaak voorkwamen en als er veel intergenerationele mobi-
liteit was tussen deze twee beroepen, dan kregen deze twee beroepen op de statusschaal
posities dicht bij elkaar. De HISCAM-schaal varieert van 1 tot 100. Technisch gezien wor-
den deze posities verkregen door het schatten van Goodmans row and column model II
(Goodman, 1979). (Prandy, 2000). Voor de schatting van de HISCAM-schaal zijn data gebruikt van 1,5 mil-
joen huwelijksaktes uit de periode 1800-1938 uit zes landen (Engeland, Canada, Frankrijk,
Duitsland, Zweden en Nederland). Als huwelijken tussen mannen en vrouwen met twee
verschillende beroepen relatief vaak voorkwamen en als er veel intergenerationele mobi-
liteit was tussen deze twee beroepen, dan kregen deze twee beroepen op de statusschaal
posities dicht bij elkaar. De HISCAM-schaal varieert van 1 tot 100. Technisch gezien wor-
den deze posities verkregen door het schatten van Goodmans row and column model II
(Goodman, 1979). 199 3. Soms zijn deze dateringen geschat, zie daarvoor het codeboek van deze dataset. Literatuur Bras, H. (2002). Zeeuwse meiden. Dienen in de levensloop van vrouwen, ca. 1850-1950. Amsterdam: Aksant. Breen, R. (2004) (red.). Social mobility in Europe. Oxford: Oxford University
Press. Brown, J.C., M.H.D. van Leeuwen & D. Mitch (2004). The history of the modern
career: An introduction. In D. Mitch, J. Brown & M.H.D. van Leeuwen (red.),
Origins of the modern career (pp. 3-41). Aldershot: Ashgate. Bulder, E. (1993). The social economics of old age. Strategies to maintain income in later
life in the Netherlands 1880-1940. Amsterdam: Thesis. Damsma, D. & J. Kok (2005). Ingedroogde harten? Partnerkeuze en sociale repro-
ductie van de Noord-Hollandse boerenstand in de negentiende en vroeg-twin-
tigste eeuw. In J. Kok & M.H.D. van Leeuwen (red.), Genegenheid en gelegenheid. Twee eeuwen partnerkeuze en huwelijk (pp. 285-307). Amsterdam: Aksant. Dehing, P.W.N.M. (1989). “... Eene soort van dynastie van spoorwegbeambten”: arbeids-
markt en spoorwegen in Nederland, 1875-1914. Hilversum: Verloren. Doff, N. (1974). Dagen van honger en ellende. Amsterdam: Meulenhoff. Erikson, R. & J.H. Goldthorpe (1992). The constant flux. A study of class mobility
in industrial societies. Oxford: Oxford University Press. Goodman, L.A. (1979). Simple models for the analysis of association in cross-
classifications having ordered categories. Journal of the American Statistical
Association, 74, 537-552. Hox, J. (2002). Multilevel analysis. Techniques and applications. Mahwah, NJ, Lon-
don: Lawrence Erlbaum Associates, Publishers. Johnson, P. (1994). The employment and retirement of older men in England and
Wales, 1881-1981. Economic History Review, 47, 106-128. Kerr, C., J.T. Dunlop, F.H. Harbison & C.A. Myers (1960). Industrialism and indus-
trial man. Cambridge, Mass.: Harvard University Press. Korupp, S. (2000). Mothers and the process of social stratification. Amsterdam, Thesis
publishers. Leeuwen, M.H.D. van (1992). Bijstand in Amsterdam, ca. 1800-1850: armenzorg als
beheersings- en overlevingsstrategie. Zwolle: Waanders. Leeuwen, M.H.D. van (2000). De eenheidsstaat: onderlinges, armenzorg en commerci- van een dubbeltje naar een kwartje? ële verzekeraars 1800-1890. Den Haag/Amsterdam: Verbond van Verzekeraars/
NEHA. ële verzekeraars 1800-1890. Den Haag/Amsterdam: Verbond van Verzekeraars/
NEHA. Leeuwen, M.H.D. van & I. Maas (1995). Sociale mobiliteit in de steden en op het
platteland. In K. Mandemakers & O. Boonstra (red.), De levensloop van de Utrecht-
se bevolking in de 19e eeuw (pp. 103-127). Assen: Van Gorcum. Leeuwen, M.H.D. van & I. Maas (1995). Sociale mobiliteit in de steden en op het
platteland. In K. Mandemakers & O. Boonstra (red.), De levensloop van de Utrecht-
se bevolking in de 19e eeuw (pp. 103-127). Assen: Van Gorcum. Literatuur 200 Leeuwen, M.H.D. van & I. Maas (2005). A short note on HISCLASS. http://historyof-
work.iisg.nl/docs/hisclass-brief.doc. Leeuwen, M.H.D. van & I. Maas (2007). Economische specialisering en verande-
rende sociale verhoudingen in de 19e en 20e eeuw. Een studie op basis van
de Nederlandse volkstellingen en huwelijksakten. In O.W.A. Boonstra, P.K. Doorn, M.P.M. van Horik, J.G.S.J. van Maarseveen en J. Oudhof (red.), Twee eeu-
wen Nederland geteld. Onderzoek met de digitale Volks- Beroeps- en Woningtellingen
1795-2001 (pp. 181-205). Den Haag: DANS en CBS. Leeuwen, M.H.D. van, I. Maas & A. Miles (2002). HISCO: Historical International
Standard Classification of Occupations. Leuven: Leuven University Press. Maas, I., P. Lambert, R. Zijdeman, K. Prandy & M.H.D. van Leeuwen (2006). HIS-
CAM. Deriving a historical occupational stratification scale. Paper gepresen-
teerd op de European Social Science History Conference, Amsterdam, 22-25
maart 2006. Maas, I. & M.H.D. van Leeuwen (2002). Industrialization and intergenerational
mobility in Sweden, Acta Sociologica, 45, 179-194. Maas, I. & M.H.D. van Leeuwen. (2004). Occupational careers of the total male
labour force during industrialization: the example of nineteenth-century
Sweden. In D. Mitch, J. Brown & M.H.D. van Leeuwen (red.), Origins of the modern
career (pp. 227-258). Aldershot: Ashgate. Maas, I. & M.H.D. van Leeuwen (2006). Over dienstboden, landarbeidsters en
andere werkende vrouwen. Beroepen van jonge vrouwen en hun moeders in
de huwelijksakten van de Zeeuwse Burgerlijke Stand. Zeeland, 15, 44-59. Mandemakers, K. (2001). De sociale structuur in Nederland rond 1900. De samen-
leving in het perspectief van de modernisering 1850-1990. In J.G.S.J. van Maar-
seveen & P.K. Doorn (red.), Nederland een eeuw geleden geteld. Een terugblik op de
samenleving rond 1900 (pp. 185-207). Amsterdam: Stichting beheer IISG. Mandemakers, K. (2004). De Historische Steekproef Nederlandse bevolking (HSN)
en het project Life Courses in Context, Bevolking en Gezin, 33, 91-114. Miles, A. (1999). Social mobility in nineteenth and early twentieth century England. Basingstoke: Macmillan. Oosterhuis, G. (1992). Van adelborst tot admiraal. De carrière van marineofficie-
ren en de dynamiek van een interne arbeidsmarkt. Mens en Maatschappij, 67,
231-254. Prandy, K. (2000). The social interaction approach to the measurement and ana-
lysis of social stratification. International Journal of Sociology and Social Policy, 19,
215-249. Priester, P.R. (1991). De economische ontwikkeling van de landbouw in Groningen, 18 Priester, P.R. (1991). De economische ontwikkeling van de landbouw in Groningen, 1800- ineke maas en marco h.d. van leeuwen 1910: een kwalitatieve en kwantitatieve analyse. Groningen: Nederlands Agrono-
misch-Historisch Instituut. Literatuur 1910: een kwalitatieve en kwantitatieve analyse. Groningen: Nederlands Agrono-
misch-Historisch Instituut. Ransom, R. & R. Sutch ( 1986). Labor of older Americans: retirement of men on
and off the job, 1870-1937. Journal of Economic History, 36, 1-30. 201 Sociaal-Democratische Studie-Club (1909). Landarbeiders. Hun arbeidsduur en
arbeidsverhoudingen. Amsterdam: Sociaal-Democratische Studie-Club. Stovel, K., M. Savage & P. Bearman (1996). Ascription into achievement: models
of career systems at Lloyds Bank, 1890-1970. American Journal of Sociology, 102,
358-399. Treiman, D.J. (1970). Industrialization and social stratification. In E.O. Lauman
(red.), Social stratification: Research and theory for the 1970’s (pp. 207-234). Indian-
apolis: University of Indiana Press. Sociale klasse, sociale mobiliteit en sterfte in
Nederland, 1850-2007 Frans van Poppel en Ruben van Gaalen Frans van Poppel en Ruben van Gaalen Ik zal nu moeten bewijzen dat de maatschappij in Engeland … de arbeiders in
een toestand gebracht heeft waarin zij niet gezond kunnen blijven en niet lang
leven kunnen; dat zij zo het leven van deze arbeiders stukje bij beetje, stapje
voor stapje ondermijnt en hen voortijdig in het graf brengt
Engels, 1976: 141 Vraagstelling In de historische literatuur wordt een somber beeld geschetst van de gezond-
heidstoestand van de arbeidende klasse in Nederland in de negentiende eeuw. Diepe armoede was een wezenlijk kenmerk van het bestaan van de grote volks-
massa, en dat had funeste consequenties voor de gezondheidstoestand van dit
deel der natie, zo wil het verhaal. Vanaf het midden van de negentiende eeuw
begonnen medici en statistici, de Nederlandse wat later dan die in Engeland en
Frankrijk, gegevens te verzamelen over verschillen in gezondheid, vooral verschil-
len in sterftecijfers, tussen sociale klassen (Davey Smith, Dorling & Shaw, 2001;
Houwaart, 1991). Hoewel over de waarde van deze indicator voor historische
populaties in het afgelopen decennium wel twijfel is gerezen (Riley, 1997; 1999),
is de gangbare opinie dat sterftegegevens geschikter zijn dan ieder ander soort
van gegevens om de ontwikkeling van de gezondheidstoestand van de bevolking
in de afgelopen twee eeuwen in kaart te brengen (Harris, 1999). Sterftegegevens
waren relatief gemakkelijk te verzamelen en vormden in de ogen van tijdgeno-
ten een ondubbelzinnige indicator van de gezondheidssituatie. Na de eerste hausse van aandacht voor de sociaal-economische sterftever-
schillen, die voortduurde tot het eerste kwart van de twintigste eeuw, trad een
periode van stilte in. De illusie bestond dat met de komst van de verzorgingsstaat
sociaal-economische sterfteverschillen waren verdwenen of op zijn minst sterk
waren afgenomen (Mackenbach, 1994). Vanaf de jaren tachtig van de vorige eeuw zijn sociaal-economische verschillen in gezondheid en sterfte in Europa echter
opnieuw een belangrijk thema van wetenschappelijk onderzoek en onderwerp
van politieke discussie geworden. Dat had veel te maken met de publicatie van
een aantal studies waarin gewag werd gemaakt van een vergroting van de sociaal-
economische ongelijkheid in sterftekansen in de jaren zestig, zeventig en tachtig
in een verscheidenheid van landen (Borrell e.a., 1999; Harding, 1995; Townsend
& Davidson, 1988; Valkonen, 1993). De vraag hoe in de loop van de tijd de sociaal-
economische ongelijkheid in sterfte zich heeft ontwikkeld, werd daardoor ook
in politiek opzicht uitermate relevant. Ook in Nederland was dat het geval. Na
eerdere weinig succesvolle pogingen van individuele onderzoekers om aandacht
voor het onderwerp te vragen, organiseerde de Wetenschappelijke Raad voor het
Regeringsbeleid samen met het ministerie van Welzijn, Volksgezondheid en Cul-
tuur in 1987 een conferentie over Gezondheidsverschillen en sociaaleconomi-
sche status (SES) (Wetenschappelijke Raad voor het Regeringsbeleid, 1987). 204 Vraagstelling Dat
resulteerde in de totstandkoming van een vijfjarig onderzoeksprogramma naar
sociaal-economische gezondheidsverschillen (Programmacommissie Sociaaleco-
nomische Gezondheidsverschillen, 1989). De eerste resultaten daarvan kwamen
in 1994 beschikbaar (Mackenbach, 1994). Ook daarna werd de trendmatige ont-
wikkeling van de SES-gerelateerde sterfte in Nederland nog herhaalde malen aan
de orde gesteld (Kunst e.a., 2004; Mackenbach e.a., 2003). 204 De groeiende belangstelling voor de temporele ontwikkeling van de relatie
tussen sociaal-economische positie en sterfte kan niet los worden gezien van een
nieuwe theoretische benadering van sociaal-economische sterfteverschillen. De
epidemiologie en het gezondheidsonderzoek richtten zich vooral op specifieke
gedragsmatige en biologische mechanismen via welke SES de gezondheid en
sterfte beïnvloedt. Daarentegen stelt de door Link en Phelan geformuleerde the-
orie van fundamental social causes (Link, Northridge, Phelan & Ganz, 1998; Link
& Phelan, 1995; Link & Phelan, 1996; 2002) dat enkele meer algemene factoren
er verantwoordelijk voor zijn dat in ieder tijdvak van de geschiedenis steeds
opnieuw specifieke maar verschillende mechanismen worden voortgebracht die
ervoor zorgen dat de negatieve samenhang tussen SES en sterfte blijft bestaan
(Lutfey & Freese, 2005). De empirische vraag of de samenhang tussen SES en sterfte inderdaad over
een lange historische periode en in verschillende ruimtelijke omgevingen dezelf-
de sterkte en richting heeft, is moeilijk te beantwoorden. Het merendeel van de
studies die informatie bevatten over trends in SES-verschillen in sterfte, heeft
een uiterst beperkte tijdshorizon. Voor de meeste landen in de westerse wereld,
met uitzondering van het Verenigd Koninkrijk, is deze informatie op zijn vroegst
pas vanaf 1960 beschikbaar. Historische studies die verder teruggaan in de tijd
waren meestal van strikt plaatselijke aard, beperkten zich meestal tot trends in
zuigelingensterfte, waren niet in staat om de sociaal-economische positie op een
consistente wijze te meten en hadden ook te lijden van ernstige methodologi-
sche tekortkomingen. frans van poppel en ruben van gaalen In dit hoofdstuk proberen we een bijdrage te leveren aan de discussie over de
langetermijntrends in sociale ongelijkheid voor de dood door gebruik te maken
van het databestand van de Historische Steekproef Nederlandse bevolking. Dit
databestand beslaat een langere periode dan veelal gebruikelijk en heeft betrek-
king op Nederland als geheel, zodat een veelheid van sociaal-ruimtelijke contex-
ten kan worden geanalyseerd. Daarnaast hanteren we verschillende definities
ter bepaling van de sociale klasse, omdat de wijze waarop de sociale klasse is
gedefinieerd effect kan hebben op de waargenomen SES-sterfteverschillen. Vraagstelling Het
kan namelijk op voorhand niet worden uitgesloten dat de uiteenlopende ken-
merken (inkomen, opleidingsniveau, sterkere affiniteit en nauw contact met
hoger geplaatste personen) waarop de klassenindelingen zijn gebaseerd, een
verschillend effect op de levensduur hebben (Kunst, 1997). Ook bestuderen we
afzonderlijk en in onderling verband de effecten op sterfte van de sociale klasse
van het gezin waaruit de onderzoekspersoon afkomstig is en die van de sociale
klasse van de onderzoekspersoon zelf. 205 Historiografie De medici en statistici die gegevens verzamelden over verschillen in sterfteni-
veau tussen sociale klassen hadden de intentie na te gaan in hoeverre de met
urbanisatie en industrialisatie samenhangende verslechtering van de sociaal-
economische positie van de arbeidende klasse tot een verhoogd sterfterisico had
geleid. Het zijn de door deze medici verrichte studies waaraan we het uitermate
negatieve beeld te danken hebben van de gezondheidstoestand van de arbeiden-
de klasse in de negentiende eeuw (Brugmans, 1975; Giele & Van Oenen, 1974;
Romijn, 1955; Van Tijn, 1977).1 Omdat de aandacht van de medici vooral uitging
naar de effecten op de gezondheid van de voor een bepaald beroep typerende
werkomstandigheden, zoals de werkhouding, de arbeidsduur, de mate van licha-
melijke inspanning, en de blootstelling aan gevaarlijke stoffen, gaat het veelal
om lokale studies onder specifieke beroepsgroepen. De negentiende-eeuwse onderzoekers maakten gebruik van weinig verfijnde
sterftematen die niet zelden ook nog eens door latere historici onjuist zijn geïn-
terpreteerd. Soms is zelfs onduidelijk hoe de onderzoekers aan hun gegevens zijn
gekomen. De Sitter (1856: 662) bijvoorbeeld stelde zonder nadere toelichting dat
‘de geneeskundige statistiek ons leert, dat de levensduur van den behoeftigen
daglooner op niet langer dan gemiddeld 32 jaren mag worden gesteld, terwijl er
bij de hoogere standen een gemiddelde levensduur van 50 jaren kan worden aan-
genomen’. De door de medicus Samuel Senior Coronel verzamelde gegevens over
de levensduur van arbeiders in een aantal sectoren van de nijverheid vormen
de belangrijkste bron waarop hedendaagse historici hun oordeel over de sterfte
van de onderlaag van de samenleving hebben gebaseerd. Het was Coronel vooral
te doen om de opsporing van gezondheidsrisico’s die verband hielden met het
beroep. Hij verrichtte onder meer onderzoek naar de sterfte onder arbeiders in sociale klasse, sociale mobiliteit en sterfte in nederland de Middelburgse calicotnijverheid (Coronel, 1861), naar de levensduur van per-
sonen die werkzaam waren in de Hilversumse (textiel)industrie (Coronel, 1862),
naar Leidse wolwerkers (Coronel, 1864b) en naar Amsterdamse diamantwerkers
(Coronel, 1864a) . Naast Coronel waren er nog andere medici (en statistici) die
empirisch onderzoek naar de verschillen in sterfte naar sociale klasse poogden
te verrichten. Van Hengel (1875) bijvoorbeeld besteedde aandacht aan de sterfte-
verschillen tussen de sociale klassen in Hilversum. 206 Voor verschillende gemeenten werden door medici ook gegevens gepu-
bliceerd over de gemiddelde leeftijd bij overlijden van de meest voorkomende
beroepen. Historiografie Daarbij valt onder meer te denken aan de studies van Broes van Dort
(1861) betreffende Goes in de periode 1830-1859, en aan die van E.C. Buchner
(1852, 3) over de sterfte in Amsterdam in de jaren 1840-1851. Buchner was ervan
overtuigd dat ‘de gemiddelde levensduur, den zo verschillende staat der gezond-
heid, der sterfte, naar de verschillende beroepen en standen bij hetzelfde volk,
in hetzelfde land, in dezelfde stad’ leren dat ‘gezondheid en levenslengte eenig
en alleen afhangt van den gunstigen of ongunstigen staat onzer levensverhou-
dingen’. Volgens Buchner leerde de sterfte van personen zonder en met beroep
dat ‘een leven zonder zorgen voor het dagelijksch onderhoud en voorzien van
de genietingen, die de wereld aanbiedt langer duurt, dan dat waar in het zweet
des aangezigts met zorgen en ontberingen het brood moet worden verdiend en
gegeten’. De Goudse geneeskundige W.F. Büchner (1842, 69) concludeerde aan de
hand van gegevens over twee begrafenissociëteiten met elk hun specifieke leden-
bestand dat ‘de sterfte hier ter steden afneemt, naarmate de ingezetenen meer
gegoed zijn’. Pas tegen het einde van de negentiende eeuw werd in Nederland op meer
wetenschappelijke wijze de aandacht gericht op de sterfteverschillen tussen soci-
ale klassen (Centraal Bureau voor de Statistiek, 1906; 1912; 1917; Centrale Com-
missie voor de Statistiek, 1898). Op basis van de overlijdensakten opgemaakt door
de ambtenaren van de burgerlijke stand werden gegevens bijeengebracht over de
leeftijd bij overlijden, het geslacht, de doodsoorzaak en het beroep van mannen,
overleden in de jaren 1891-1895. Later werden ook gegevens gepubliceerd over
de periodes 1896-1900, 1896-1903 en 1908-1911. De betreffende statistieken zijn
onderling moeilijk vergelijkbaar door afwijkende leeftijds- en beroepsindelin-
gen. De statistische gegevens werden in de tijd van publicatie niet aan analyses
onderworpen, maar later hebben auteurs wel geprobeerd dat te doen (De Bie,
2006; Van Reek, 1985; Van Reek, 1993). Langetermijntrends in SES-gerelateerde sterfte: was er een klas
sengradiënt, was er sprake van convergentie of van divergentie? Informatie over historische trends in sociaal-economische sterfteverschillen is
van essentiële betekenis voor het huidige soms verhitte debat over een moge-
lijke toename van deze sterfteverschillen sinds de jaren zestig (Whitehead, frans van poppel en ruben van gaalen 1998). Alleen met eerdere sterftemetingen is het mogelijk om te beoordelen
of sociale verschillen werkelijk zijn toe- of afgenomen. Die eerdere metingen
moeten bij voorkeur meerdere decennia beslaan. Sterfteprocessen zijn lange-
termijnprocessen en veel factoren die sterfterisico’s beïnvloeden, kennen een
lange vertragingsperiode (roken, drinken, voeding). De processen die de sociale
sterfteverschillen vormgeven (gezondheidspolitiek, inkomensbeleid etc.) heb-
ben eveneens een zeer lange adem. Alleen met deze trendinformatie kan de
centrale hypothese van Link en Phelans fundamental social causes-theorie worden
getoetst. 207 Uitgangspunt van deze theorie, die de afgelopen jaren sterk in de belangstel-
ling is gekomen (Link e.a., 1998; Link & Phelan, 1995; Link & Phelan, 1996; 2002;
Phelan, Link, Diez-Roux, Kawachi & Levin, 2004), is dat verschillen in sterfte naar
SES over een lange periode en op een veelheid van plaatsen zijn terug te vinden
(voor een enigszins vergelijkbare opvatting over de historische onvermijdbaar-
heid van sociale-klasseverschillen in sterfte, gegrondvest op de onmogelijkheid
richting te kunnen geven aan het eigen leven, zie Marmot, 2004). In die zin is
de theorie in tegenspraak met de opvatting van Antonovsky, die meent dat deze
verschillen door de tijd heen sterk kunnen variëren en in bepaalde periodes ook
afwezig zijn geweest. Link en Phelan stellen daar bijvoorbeeld tegenover dat
‘wat ook op enig moment het ziekten- en risicoprofiel is, steeds zullen personen
die betere toegang hebben tot relevante sociale en economische hulpbronnen
minder door ziekte getroffen worden’ (Link & Phelan, 1996, 472). Elders stellen
Link e.a. (1998, 375-376) dat het ‘tussen 1840 en 1995 blijven voortbestaan van de
samenhang tussen SES en sterfte een feit is dat verklaring behoeft’ en beweren
Phelan e.a. (2004) dat tussen de negentiende eeuw en het heden ‘sociaal-econo-
mische ongelijkheid in de sterfte onveranderd is gebleven’. Dat de verbetering
van de volksgezondheid en de toename van de levensverwachting niet tot een
vermindering van de SES-verschillen in sterfte hebben geleid, verklaren Link en
Phelan (1996, 472) doordat sociale condities die betrekking hebben op hulpbron-
nen als kennis, geld, macht, prestige, en sociale relaties grote invloed hebben
op de mogelijkheid van mensen om gezondheidsrisico’s te vermijden en om de
negatieve gevolgen van ziekten, wanneer die hebben toegeslagen, te minimali-
seren. Langetermijntrends in SES-gerelateerde sterfte: was er een klas
sengradiënt, was er sprake van convergentie of van divergentie? Vanwege de ruime inzetbaarheid van deze sociale en economische hulp-
bronnen, staan deze fundamentele factoren via een grote verscheidenheid aan
risicofactoren met een veelheid van gezondheidsuitkomsten in verband. Omdat
deze hulpbronnen in uiteenlopende situaties op verschillende wijzen ingezet
kunnen worden beïnvloeden deze fundamentele factoren de ziektelast ook dan
wanneer het patroon van de risicofactoren wezenlijk verandert. Terwijl in de negentiende eeuw ondermaatse huisvesting, hoge woondicht-
heid en het ontbreken van riolering en waterleiding tot hogere sterfte aan infec-
tieziekten onder de lagere sociale klassen aanleiding gaven, waren het in de jaren
vijftig en zestig van de twintigste eeuw de hogere tabaksconsumptie, minder
beweging, eenzijdige ongezonde voeding en een geringer beroep op de gezond- sociale klasse, sociale mobiliteit en sterfte in nederland heidszorg die ertoe leidden dat de lagere sociale klassen een hogere sterfte ken-
den dan de hogere lagen. De fundamental causes theory is verschillende malen getoetst op hedendaagse
gegevens maar de vraag of SES-verschillen in sterfte over een langere historische
periode en op een verscheidenheid van plaatsen kunnen worden waargenomen,
is niet op bevredigende wijze beantwoord. Voor de negentiende eeuw hebben
Link en Phelan niet meer dan anekdotische informatie en resultaten van enkele
Engelse lokale studies weten te presenteren die geen beeld geven van de sterkte
van de samenhang tussen SES en sterfte. Link, Northridge, Phelan en Ganz (1998)
stellen terecht dat ‘een toets van het idee van het bestaan van fundamental causes
idealiter gebaseerd zou moeten zijn op een groot bestand met historische gege-
vens over individuen en hun ziektelast’. Het grote probleem is dat de beschikbare
informatie over trends in SES-sterfteverschillen op nationale schaal een beperk-
te tijdshorizon heeft. Voor de VS bijvoorbeeld bestaan slechts gegevens over de
periode vanaf 1960, hoewel ook voor de Tweede Wereldoorlog de belangstelling
voor het onderwerp daar groot was; zie Krieger en Fee (1996). Onze kennis van
de samenhang tussen SES en sterfterisico’s leunt daardoor zwaar op Engelse
gegevens. De informatie over de langetermijntrends in SES-sterfteverschillen in
het Verenigd Koninkrijk is beschikbaar vanaf de jaren negentig van de negen-
tiende eeuw en met beperkingen zelfs vanaf de daaraan voorafgaande jaren zes-
tig (Pamuk, 1985; Rogers Hollingsworth, 1981; Woods, 2004). Er zijn duidelijke
aanwijzingen dat het niet mogelijk is om de in Engeland waargenomen SES-
sterfteverschillen te generaliseren naar andere landen. Langetermijntrends in SES-gerelateerde sterfte: was er een klas
sengradiënt, was er sprake van convergentie of van divergentie? De aard en de omvang
van de problemen die de overgang naar een industriële samenleving met zich
meebracht, verschilden daarvoor te zeer van die in andere landen: de industri-
alisatie kwam er eerder op gang, de bevolkingsgroei vond vooral plaats in grote
industriesteden, de klassentegenstellingen waren er sterker, de sociale klassen
homogener en sociale wetgeving ter beteugeling van de problemen kwam eerder
van de grond (Preston & Haines, 1991). 208 De heersende opvattingen met betrekking tot de langetermijntrends in SES-
gerelateerde sterfte zijn gebaseerd op een studie van Aaron Antonovsky (1967). Antonovsky stelde, op basis van veelal rudimentaire gegevens betrekking heb-
bend op de periode vóór 1900 en afkomstig uit een veelheid van plaatsen dat
in de westerse wereld drie opeenvolgende fasen te onderkennen zijn in de lan-
getermijnontwikkeling van de sterfteverschillen naar sociale klasse. Tot circa
1650 werden volgens hem geen substantiële verschillen in levensverwachting
aangetroffen. Tussen 1650 en 1850 ontstond er op het moment dat een industrie-
proletariaat tot ontwikkeling kwam, een groeiende kloof in levensverwachting,
die toe te schrijven valt aan een verbetering van de positie van de midden- en
bovenste lagen en een geringere toename of zelfs een afname van de levensver-
wachting in de lagere strata. Na 1850 verkeerde deze tendens in het tegendeel en
werd de kloof tussen de sociale klassen kleiner, zodat in de jaren zestig van de
twintigste eeuw de verschillen geringer waren dan ze in de eeuwen van 1600 tot
1850 ooit waren geweest. frans van poppel en ruben van gaalen Het model van Antonovsky is door verschillende auteurs op basis van even-
eens beperkte gegevens nader gepreciseerd. Woods en Williams (1995) gebruik-
ten daarvoor nieuwe gegevens over prosopografische studies van de sterfte onder
de aristocratie en de middenklasse in Engeland, Frankrijk en Zwitserland, onder
meer afkomstig van het werk van Alfred Perrenoud. In hun opvatting was al vóór
1650 sprake van sociale-klasseverschillen in sterfte, maar was van een duidelijke
gradiënt geen sprake. Rond 1750 nam de elite haar verwachte positie in en had
deze een hogere dan gemiddelde levensverwachting. Dit verschil bleef gehand-
haafd tot de twintigste eeuw. Woods en Williams benadrukken dat er vanaf het
moment dat sociale-klasseverschillen zichtbaar werden sprake was van periodes
van divergentie en convergentie. In een latere studie vergeleek Woods (2004) de
levensverwachting van elites in een verscheidenheid van Europese landen met
die van de bevolking van Engeland als geheel. Langetermijntrends in SES-gerelateerde sterfte: was er een klas
sengradiënt, was er sprake van convergentie of van divergentie? De conclusie was dat in de negen-
tiende eeuw sprake was van een vorm van sociale-klassesterfteverschillen, maar
dat dat vóór die tijd niet duidelijk het geval was. De Engelse ervaringen met
betrekking tot de periode 1860-1910 wezen uit dat de sociale-klassengradiënt
in sterfte een wezenlijk twintigste-eeuws fenomeen was; vóór die tijd speelde
de ruimtelijke omgeving in de brede zin van het woord (de waterkwaliteit, de
bodemtoestand), een veel bepalender rol dan de sociale positie. Ongeacht de
sociale klasse waartoe men behoorde werden de sterftekansen vooral door deze
voor ieder individu gelijke risicofactoren bepaald. Razzell en Spence (2006) bewe-
ren zelfs, op basis van een grote variëteit aan gepubliceerde kleinschalige stu-
dies en nieuw verzamelde lokale data voor Engeland dat ‘een samenhang tussen
sociaaleconomische status en sterfte op volwassen leeftijd pas in de twintigste
eeuw valt vast te stellen’. Blum, Houdaille en Lamouche (1990) echter vonden
voor Frankrijk vanaf het midden van de achttiende eeuw hogere waarden van
de levensverwachting op twintigjarige leeftijd voor personen die tot de hogere
sociale klassen behoorden dan voor hen die tot de middenklasse konden worden
gerekend; op hun beurt waren die weer beter af dan losse en vaste arbeiders. 209 Historici in een groot aantal landen hebben in het afgelopen decennium
getracht een nieuwe impuls te geven aan de studie van de SES-verschillen in
sterfte door op basis van de originele bronnen van de burgerlijke stand nieuwe
schattingen te maken van het sterfteniveau van de verschillende sociale klassen. Daarbij werd van uiteenlopende benaderingen gebruikgemaakt. In sommige stu-
dies werden de gegevens van de burgerlijke stand direct op individueel niveau
gekoppeld aan de gegevens van de ‘population at risk’ ten tijde van een volkstel-
ling. In andere werden geaggregeerde aantallen sterfgevallen gekoppeld aan de
risicobevolking, terwijl weer andere auteurs een prospectieve benadering volg-
den (Kunst, 1997). Voorbeelden zijn te vinden voor België (Backs, 2001; Neven,
2000; Oris, 1998); voor Italië en Zweden (Bengtsson, 2004; Breschi, Derosas & Man-
fredini, 2004; Tsuya & Nystedt, 2004) en voor de VS (Currie & Stabile, 2002; Ferrie,
2003). Ook deze studies beperkten zich echter tot een enkele gemeenschap en
besteedden geen aandacht aan veranderingen in de relatie tussen SES en sterfte. Langetermijntrends in SES-gerelateerde sterfte: was er een klas
sengradiënt, was er sprake van convergentie of van divergentie? sociale klasse, sociale mobiliteit en sterfte in nederland Voor Frankrijk (Blum e.a., 1990; Bourdieu & Kesztenbaum, 2004) en Nederland
(Van Poppel, Deerenberg, Wolleswinkel-van den Bosch & Ekamper, 2005) werden
weliswaar schattingen van historische trends in de levensverwachting naar soci-
ale klasse gemaakt (op basis van historische longitudinale gegevens), maar deze
studies waren gebaseerd op beperkte aantallen waarnemingen. Omdat deze en
andere historische studies geen informatie verschaffen over de twintigste-eeuwse
ontwikkeling, hebben zij ook niet hun weg gevonden binnen het gedachtegoed
van de epidemiologie. 210 Sociale-klasseverschillen in sterfte: uitgangspunten Verschillende
indelingen kunnen dus ook tot een ander patroon van sociale-sterfteverschillen
leiden (Craig & Forbes, 2005). 211 Een tweede punt betreft de noodzaak de ruimtelijke context in de analyse
te betrekken. In het hedendaagse epidemiologische onderzoek is de aandacht
sterk gericht op de invloed die omgevingskenmerken – zoals de gemeenschaps-
organisatie en de aanwezigheid van gezondheidsvoorzieningen – kunnen uitoe-
fenen op het sterfterisico van individuen, onafhankelijk van de kenmerken van
die individuen (Krieger, Williams & Moss, 1997; Kunst, 1997; Robert, 1999; Yen &
Syme, 1999). Dat besef is ook doorgedrongen in het historisch onderzoek. Woods
en Williams (1995) wezen erop dat de ruimtelijke omgeving een factor is die ook
verstorend kan werken op de geobserveerde historische SES-sterfteverschillen,
tot ver in de negentiende eeuw.2 Johansson en Kasakoff (2000) benadrukken dat
tot in de eerste decennia van de twintigste eeuw de disease environment en econo-
mische omstandigheden van plaats tot plaats enorm varieerden. Dat kon leiden
tot grote verschillen tussen regio’s in de gemiddelde levensduur, in Nederland
bijvoorbeeld in het midden van de negentiende eeuw tot in de orde van groot-
te van tien jaar. Historische sociale-klasseverschillen in sterfte reflecteren in
belangrijke mate de invloed van ruimtelijke kenmerken, zoals de kwaliteit van
het drinkwater en de aanwezigheid van riolering (zie ook Spree, 1988, 89-90). Er is een derde aspect dat in hedendaagse epidemiologische studies naar
SES-gerelateerde sterfte prominent aanwezig is, maar dat in het historische
onderzoek niet aan de orde wordt gesteld, te weten het belang van de sociaal-eco-
nomische positie van het gezin van oorsprong versus dat van de eigen positie. Er
is een groeiende hoeveelheid literatuur die uitwijst dat de sterfte op volwassen
leeftijd, vooral de sterfte aan hart- en vaatziekten, niet alleen samenhangt met
risicofactoren die direct verband houden met de sociaal-economische positie op
volwassen leeftijd, maar ook met factoren die terugverwijzen naar de sociaal-
economische positie van het gezin waarin men opgroeide (Davey Smith, Blane &
Bartley, 1994; Lundberg, 1991; 1993; Power e.a., 2007; Wadsworth, 1986). De met
de sociale klasse verbonden risico’s op jonge leeftijd kunnen versterkt of gecom-
penseerd worden door de risico’s die samenhangen met de sociale klasse waar-
toe men als volwassene behoort (Power e.a., 2007). In het recente onderzoek is
de nadruk gelegd op het cumulatieve effect van de sociaal-economische positie
over de levensloop. Dat impliceert dat hoe langer de persoon is blootgesteld aan
ongunstige omstandigheden, des te groter de gezondheidsrisico’s zijn. Sociale-klasseverschillen in sterfte: uitgangspunten In verschillende opzichten zijn de historische studies van de langetermijnontwik-
keling in SES-sterfteverschillen niet goed in staat om de vragen te beantwoorden
die in het hedendaagse epidemiologische onderzoek centraal staan. Dat is voor
een deel het gevolg van het ontbreken van de daarvoor noodzakelijke historische
informatie, maar voor een deel van de gescheiden circuits waarin historici en
epidemiologen zich bewegen. We wijzen hier op een viertal punten. Opleiding en de op basis van het beroep bepaalde sociale klasse vertonen
in hedendaags onderzoek een sterke samenhang met sterfte. Beide indicatoren
van SES hangen sterk samen, maar meten ook verschillende aspecten. Het oplei-
dingsniveau reflecteert meer de jeugdomstandigheden van betrokkenen, de fase
waarin de basis wordt gelegd voor gedragingen op het terrein van de gezond-
heid waarvan het effect lang voelbaar is. Het beroep hangt vooral samen met de
ervaringen en risico’s van het volwassen leven en reflecteert sterker de materiële
situatie van de betrokkenen (Martikainen, Blomgren & Valkonen, 2007). In epi-
demiologische studies wordt duidelijk de voorkeur gegeven aan het opleidings-
niveau, terwijl in het historisch onderzoek (noodgedwongen) gebruik wordt
gemaakt van het beroep als maatstaf voor de sociale-klassepositie. g
p
p
De keuze voor het beroep heeft vooral praktische redenen, maar valt ook
op grond van theoretische overwegingen wel te verdedigen. In de periode vóór
1960 is informatie over het opleidingsniveau slechts in bescheiden mate beschik-
baar. Bovendien maakt het slechts een beperkte mate van differentiatie mogelijk
door de oververtegenwoordiging van de groep met uitsluitend lager onderwijs. Van belang is ook dat de met het opleidingsniveau samenhangende verschil-
len in voor de gezondheid relevante gedragspatronen in historische populaties
een minder grote rol speelden dan de met het beroep samenhangende factoren. We volgen hier dan ook de opvatting van Davey Smith e.a. (1998) dat de op het
beroep gebaseerde indeling naar sociale klasse sterker differentieert dan oplei-
ding. Wel is van belang te beseffen dat de wijze waarop de sociale klasse is gede-
finieerd effect kan hebben op de waargenomen SES-sterfteverschillen. Hoewel
het gebruik van verschillende klassenindelingen over het algemeen dezelfde ten-
densen aan het licht brengt, kan de precieze omvang van de veranderingen in
sociaal-economische ongelijkheid variëren al naar gelang de toegepaste indeling. frans van poppel en ruben van gaalen Evaluaties van meerdere sociale klassenschema’s hebben aan het licht gebracht
dat ‘klasseneffecten’ voor een grote verscheidenheid van uitkomsten kunnen
verschillen al naar gelang de gekozen indeling (Leye & Joye, 1994). Sociale-klasseverschillen in sterfte: uitgangspunten Een alter-
natieve hypothese is dat ervaringen in het verdere verleden in betekenis vermin-
deren naarmate de tijd verstrijkt. Gegeven de complexiteit van de relatie tussen
SES in de kindertijd, gezondheid-gerelateerde selectie op SES, en SES op volwas-
sen leeftijd enerzijds en sterfte anderzijds bepleit Mare een benadering waarin
de sterfte op volwassen leeftijd wordt gezien als een functie van vroegere en hui- sociale klasse, sociale mobiliteit en sterfte in nederland dige SES en empirisch gepoogd wordt het gewicht van beide posities te bepalen
(Mare, 1990). Nauw verwant aan de hier besproken thematiek is de vraag in hoe-
verre sociale mobiliteit en sterfte samenhangen. Terwijl de sociaal-economische
positie veelal wordt beschouwd als een determinant van slechte gezondheid en
hogere sterfte, kan omgekeerd een slechte gezondheidstoestand ook de uiteinde-
lijk te bereiken sociaal-economische positie bepalen. Onderzoek naar de vraag of
de gezondheidssituatie invloed uitoefent op de kans een bepaalde sociale positie
te bereiken, heeft weinig consistente resultaten opgeleverd. Recente literatuur
wekt zelfs de suggestie dat met de gezondheid samenhangende sociale mobi-
liteit geen vergroting, maar juist een verkleining van de SES-sterfteverschillen
veroorzaakt, omdat de opwaarts mobiele personen in een slechtere gezondheid
verkeren (Manor, Matthews & Power, 2003). 212 Een vierde punt betreft de afwezigheid in veel van het hedendaagse onder-
zoek van informatie over SES-verschillen in sterfte bij volwassen vrouwen. Naarmate er in het epidemiologisch onderzoek meer gebruik wordt gemaakt
van opleidingsniveau als indicatie voor SES, wordt het gebruikelijker ook SES-
verschillen in sterfte onder vrouwen te rapporteren. Voorheen werd hiervoor de
sociale klasse van de echtgenoot gepresenteerd. Op basis van het eigen beroep
werden voor vrouwen over het algemeen minder sterke verschillen in SES-gerela-
teerde sterfte gevonden dan bij mannen. Deels hangt dat samen met het relatief
grote aantal vrouwen dat geen betaalde arbeid verricht, waardoor het op beroep
gebaseerde sociale-klassecriterium minder informatief is. Deels hangt het ermee
samen dat opleiding, de drijvende kracht achter de uiteindelijk door de vrouw
bereikte sociale klasse, bij vrouwen minder sterk samenhangt met gezondheid,
deels met de minder zware fysieke eisen die aan het beroep van vrouwen worden
gesteld (Cambois, 2004). De hier gepresenteerde studie van de langetermijnontwikkeling van de
sociale-klasseverschillen in sterfte tracht met de hier besproken uitgangspunten
rekening te houden. Onze studie heeft een aantal onderscheidende kenmerken
die het mogelijk maken de tekortkomingen van eerdere studies te voorkomen. 212 Sociale-klasseverschillen in sterfte: uitgangspunten We bestuderen gegevens die betrekking hebben op een lange tijdsperiode, gedu-
rende welke Nederland radicale veranderingen in de economische en sociale
structuur onderging (toename inkomen, industrialisatie en urbanisatie). Het
betreft de geboortegeneraties 1850-1922, waarin de levensverwachting bij man-
nen op de drempel van de volwassenheid toenam van 44,3 naar 54,8 jaar en bij
vrouwen van 45,2 naar 61,8 jaar. We bestuderen gegevens die betrekking heb-
ben op het land als geheel, en kunnen dus rekening houden met de situatie
in een veelheid van ecologische, sociale en economische omstandigheden, met
plattelandsgebieden en kleine en grote steden. De door ons gebruikte gegevens
op microniveau maken het mogelijk de sociale klasse van individuen zowel op het
moment van geboorte als op volwassen leeftijd te bepalen, terwijl daarnaast ook
de sociale klasse waarin vrouwen geboren worden of door huwelijk in terechtko-
men kan worden vastgesteld. Bovendien hanteren we twee verschillende indelin- frans van poppel en ruben van gaalen gen naar sociale klasse, zodat we kunnen nagaan of het de specifieke indeling is
die de uitkomsten bepaalt. We richten ons op de sterfte van die personen die ten
minste achttien jaar oud werden. Vanaf deze leeftijd werd door het merendeel
van de bevolking een beroep uitgeoefend, zijn personen dus zelfstandig in een
sociale klasse onder te brengen. 213 Data Friesland, sociale klasse, sociale mobiliteit en sterfte in nederland Groningen en Zeeland behoorden tot de concentrische cirkel van productieve
agrarische gebieden direct rondom de economische kern en worden samengevat
als de groep Noordwest. Zeeland had weliswaar tot circa 1880 een duidelijk hoge-
re kindersterfte dan de beide andere provincies, maar aangezien we ons richten
op de sterfte op volwassen leeftijd van geboortecohorten 1850 en later, vormt dit
niet echt een probleem.3 De provincies Noord-Brabant, Limburg en Gelderland,
Overijssel en Drenthe vallen onder de meer perifere categorie Zuidoost. Hier was
de levensverwachting bij aanvang van de periode redelijk hoog. Later trad een
relatieve verslechtering op in de positie van deze provincies. 214 De afzonderlijke gemeenten zijn geclassificeerd in urbane en rurale gemeen-
ten op basis van het inwonertal. Eerder onderzoek heeft uitgewezen dat in Neder-
land – net zoals elders – de mate van stedelijkheid in de negentiende eeuw een
duidelijk verhogend effect op de sterftekansen had, terwijl in een latere fase de
sterfte in meer verstedelijkte gebieden juist lager lag dan in rurale gebieden (Van
Poppel, 1989). Om de veranderingen in de tijd te kunnen weergeven onderscheiden we drie
geboortegeneraties: de eerste is geboren tussen 1850 en 1879 en valt min of meer
samen met de groep die de eerste fase van de stijging van de levensverwachting
meemaakte (de levensverwachting op twintigjarige leeftijd nam toe van 43 naar
49 jaar bij mannen en van 44 naar 50 jaar bij vrouwen), de tweede generatie,
geboren tussen 1880 en 1899, ervaart een verdere toename van zeven jaar bij de
mannen en 5,5 jaar bij de vrouwen, de derde generatie (1900-1922) wordt gecon-
fronteerd met een toename van de sterfte aan chronische ziekten waardoor de
levensverwachting nauwelijks nog toeneemt (met twee jaar bij mannen en 4,5
jaar bij vrouwen). Om de sociale klasse te bepalen is gebruikgemaakt van de beroepsaandui-
dingen uit de geboorteakten, de huwelijksakten en het bevolkingsregister. De
sociale klasse in het gezin van oorsprong is bepaald op basis van het beroep van
de vader van het kind zoals vermeld in de geboorteakte. Het beroep op volwassen
leeftijd is bepaald op basis van het eerst vermelde beroep in het bevolkingsre-
gister c.q. de huwelijksakte, van de persoon zelf indien de onderzoekspersoon
een man was, en van de echtgenoot van de vrouw als de onderzoekspersoon een
vrouw was. Data Er wordt gebruikgemaakt van de Historische Steekproef Nederlandse bevolking
(HSN). De HSN is een nationaal databestand waarin informatie is opgenomen
over de levensloop van een groot aantal personen geboren in Nederland in de
jaren 1812-1922 (Mandemakers, 2000). We analyseren twee subsets uit deze data. De dataset ESM release ESM.08, September 2006 (n = 7.856) bevat gegevens uit de pro-
vincies Utrecht, Zeeland en Friesland voor de geboorteperiode 1850-1922. Deze
set werd voor de andere provincies aangevuld met de dataset HSN Levenslopen,
release 2007.01, zie ook de bijdrage van Mandemakers in dit boek. Deze betreft
onderzoekspersonen uit de geboorteperiode 1883-1922 (n = 8.427). Alleen voor de
generaties die na 1849 zijn geboren, is bepaalde belangrijke informatie, zoals de
godsdienst, beschikbaar. De uitkomstvariabele waarin we zijn geïnteresseerd is de leeftijd bij overlij-
den. Informatie over de datum van overlijden is afkomstig van de overlijdens-
akten, van de vermelding van het overlijden in het bevolkingsregister of op de
gezinskaart, en van de persoonskaarten en persoonslijsten van de Gemeentelijke
Basisadministratie. Deze beide laatste bronnen zijn per definitie alleen beschik-
baar voor overleden onderzoekspersonen. Over onderzoekspersonen die bij het
aanmaken van een persoonskaart eind jaren dertig nog in leven waren, maar tot
op heden nog niet zijn overleden is na 1939 veelal geen informatie meer open-
baar en door de HSN verzameld. Dat heeft belangrijke consequenties voor de
vaststelling van de status van de onderzoekspersonen. Van de 16.283 onderzoekspersonen is van 2.379 geen overlijdensdatum
bekend, noch een laatste uitschrijfdatum uit het bevolkingsregister. Deze kun-
nen niet in de analyse worden opgenomen. Van de overgebleven 13.904 personen
zijn er op achttienjarige leeftijd nog 9.838 in leven. Van deze 9.838 is voor 1.871
personen (19,0 procent) geen overlijdensdatum bekend, maar wel een laatste uit-
schrijfdatum. De gegevens van de provincies zijn samengevoegd in drie regio’s op basis
van de ontwikkeling van de levensverwachting in de jaren 1840-1851, 1901-1902,
1956-1960 en 1990-1991 (Van Poppel & Beekink, 2003) en op basis van geografische
nabijheid en vergelijkbare economische omstandigheden. De provincies Utrecht
en Noord- en Zuid-Holland, het economische en culturele centrum van het land,
werden tot het laatste kwart van de negentiende eeuw door hoge sterfte geka-
rakteriseerd. Vanaf circa 1880 nam de levensverwachting hier echter sterker toe
dan elders. We voegen de drie provincies samen tot de categorie West. Data Gebruik is gemaakt van het HISCO-coderingsschema voor beroeps-
titels (Historical International Standard Classification of Occupations) (Van Leeuwen,
Maas & Miles, 2002). Uitgangspunt van HISCO zijn de taken die aan een bepaalde
historische beroepstitel verbonden zijn. Op basis van de HISCO-codes zijn vervolgens twee sociale klassenindelin-
gen gemaakt: de SOCPO-indeling voorgesteld door Van de Putte en Miles (2005),
en de HISCLASS-indeling, voorgesteld door Van Leeuwen en Maas (2005).4
De SOCPO-(Social Power-)indeling heeft als leidend principe ‘sociale macht’,
omschreven als de ‘objectief’ vaststelbare mogelijkheden van een persoon om
de eigen levenskansen te beïnvloeden op basis van de controle over schaarse
hulpmiddelen. Sociale macht is gebaseerd op twee bronnen: materiële (econo- frans van poppel en ruben van gaalen mische) en niet-materiële (culturele) macht. Op basis van de samenvoeging van
de economische en culturele machtsdimensies zijn vijf groepen onderscheiden. Op het hoogste niveau (niveau 5) zitten personen die algemene beleidstaken uit-
voeren, bezitters en bedrijfshouders op macroniveau, hooggeschoolde niet-hand-
arbeiders en personen die tot de adel behoren; op niveau 4 degenen die supervisie
uitoefenen over geschoolde werknemers, bezitters die op mediumschaalgrootte
werkzaam zijn, supergeschoolde handarbeiders en geschoolde niet-handarbei-
ders; op niveau 3 personen die supervisie uitoefenen over laag- of ongeschoolden
en de geschoolde handarbeiders; op niveau 2 de bezitters op microschaalgrootte
en laaggeschoolden, en op niveau 1 de ongeschoolden. Vanwege kleine aantallen
zijn de niveaus 4 en 5 samengevoegd. We duiden de opeenvolgende groepen aan
als de boven-/middenklasse (niveaus 4 en 5), de geschoolde arbeiders, de laag-
geschoolden en de ongeschoolden. We voegen een vijfde groep toe waarvan het
beroep onbekend is. 215 Het schema van Van Leeuwen en Maas is gebaseerd op vier dimensies: het
onderscheid tussen handarbeid en niet-handarbeid, het niveau van geschoold-
heid, de mate waarin men supervisie over anderen uitoefent en de sector van
de economie. In HISCLASS worden twaalf sociale klassen onderscheiden, die om
te geringe aantallen waarnemingen te voorkomen zijn samengevoegd tot zes
klassen: managers, professionals, kantoor- en handelsbedienden; voorlieden en
geschoolde arbeiders; boeren en vissers; laaggeschoolde arbeiders; ongeschoolde
arbeiders; landarbeiders (laaggeschoold en ongeschoold). Ook hier voegen we
een groep toe waarvan het beroep onbekend is. We definiëren opwaartse sociale mobiliteit volgens de HISCLASS-indeling als
een verbetering van ten minste twee stappen voor personen van wie de vader
behoorde tot de ongeschoolden. Data We nemen voor deze analyse aan dat laagge-
schoolde landarbeiders tot een lagere klasse behoren dan laaggeschoolde arbei-
ders buiten de landbouw, aangezien de gemiddelde levensverwachting van alle
kinderen uit de eerste groep gemiddeld 2,5 jaar lager ligt dan die van kinderen
in de tweede groep. Volgens de SOCPO-indeling gaat het om personen die oor-
spronkelijk behoorden tot de groep van ongeschoolden en halfgeschoolden en
minstens twee stappen zijn geklommen.5 Tot slot nemen we religie als controlevariabele in de analyse mee. In Neder-
lands historisch onderzoek (Van Poppel, Schellekens & Liefbroer, 2002) en in bui-
tenlandse studies (zie onder veel meer Hummer, Rogers, Nam & Ellison, 1999;
Levin, 1994) is gewezen op de hogere sterfterisico’s van katholieken ten opzichte
van protestanten en joden. Beschrijvende kenmerken van de studie Tabel 1 geeft een overzicht van de belangrijkste beschrijvende kenmerken
van de steekproef voor alle in het onderzoek opgenomen personen. In eerste
instantie is het van belang na te gaan of de HSN-data representatief zijn voor sociale klasse, sociale mobiliteit en sterfte in nederland het sterftepatroon van de Nederlandse bevolking als geheel. Weliswaar is voor
een groot deel van de geselecteerde onderzoekspersonen een overlijdensdatum
gevonden; dat biedt nog geen garantie dat de uitkomstvariabele, de leeftijd bij
overlijden, op accurate wijze is te meten. Van bijna eenvierde deel van het meest
recente cohort is de overlijdensdatum (nog) niet bekend. Tabel 2 geeft voor ver-
schillende kenmerken het aandeel waarvan geen overlijdensdatum werd gevon-
den (19 % van het totaal). Uit de tabel blijkt dat er voor de controlevariabelen
geslacht, stedelijkheid en regio van de geboortegemeente weinig verschillen zijn
in het percentage personen voor wie uiteindelijk geen overlijdensdatum kon
worden vastgesteld. Van belang is vooral dat ook per sociale klasse de verschil-
len gering zijn, ongeacht de indeling, of het nu gaat om de sociale klasse in het
gezin van oorsprong, die van de persoon zelf of die van de echtgenoot. Opvallend
is wel het relatief hoge aandeel van personen zonder een overlijdensdatum voor
die personen/echtgenoten, die geen beroep hadden of van wie het beroep niet
kon worden vastgesteld. 216 Een tweede probleem is dat van de meest recente geboortegeneraties nog
een deel in leven is, waardoor deze onderzoekspersonen niet verder konden wor-
den gevolgd in de bevolkingsregisters dan tot de invoering van de persoonskaart
(1938-1939). Zijn van de betrokkenen geen persoonskaarten of persoonslijsten
uit de Gemeentelijke Basisadministratie gevonden, dan kunnen de betreffende
personen bij het afsluiten van dit gedeelte van de dataverzameling (2005) nog
in leven zijn. Voor deze personen zou als datum van censureren het tijdstip van
laatste waarneming in de jaren dertig kunnen worden gebruikt.6 De methode
van dataverzameling houdt echter in dat er in de meest recente geboorteco-
horten een grotere kans is om informatie te verzamelen over een relatief vroeg
overledene dan over een relatief laat overledene. Dat geldt vooral voor de meest
recente cohorten, maar deze tendens bestaat in lichtere mate ook bij de oudere
geboortecohorten. De ontbrekende data zijn niet random verdeeld, maar han-
gen samen met het proces dat wordt bestudeerd, i.c. de sterfte (Blossfeld & Roh-
wer, 2002). Beschrijvende kenmerken van de studie Dat zou tot een overschatting van het feitelijke sterfterisico leiden.7
Om dit te voorkomen wordt ervoor gekozen om uit te gaan van de aanname dat
personen die na 1910 zijn geborenen en van wie geen overlijdensdatum bekend
is en die tot 1938 in het bevolkingsregister konden worden gevolgd, in 2005 nog
in leven zijn. De effecten van de methode van dataverzameling worden zichtbaar wanneer
we het afstervingsproces van de verschillende HSN-cohorten vergelijken met sterf-
tegegevens voor bijna identieke geboortegeneraties voor Nederland als geheel,
gebaseerd op CBS-cijfers (figuur 1).8 Op zich volgen de HSN-cohorten in grote lijnen
het verwachte patroon, dat wordt gekenmerkt door een verhoging van de overle-
ving in de successievelijke cohorten. Wel is er een relatieve verslechtering in het
cohort 1900-1922 ten opzichte van het cohort 1880-1899, die zonder twijfel voor een
belangrijk deel moet worden toegeschreven aan de gevolgde methode van data-
verzameling. Na de veertigjarige leeftijd is echter sprake van een duidelijke verbe- frans van poppel en ruben van gaalen sociale klasse, sociale mobiliteit en sterfte in nederla
Tabel 1 Beschrijvende kenmerken van de HSN-steekproef (N=9.838)
Proportie/M
Min
Max
SOCPO-indeling
Proportie HISCLASS-indeling
Proport
Leeftijd bij overlijden
67,87
18
104
Vader
Vader
Periode van geboorte
-
-
Laag-/ongeschoolde arbeiders landbouw
0,13
1850-1879
0,20
Ongeschoolde arbeiders 0,33
Ongeschoolde arbeiders
0,21
1880-1899
0,33
Laaggeschoolde arbeiders 0,11
Laaggeschoolde arbeiders
0,11
1900-1922
0,47
-
-
Boeren en vissers
0,15
Geslacht
Geschoolde arbeiders
0,19
Voorlieden en geschoolde arbeiders
0,17
Mannen
0,51
Midden-/bovenklasse
0,31
Managers en professionals
0,16
Vrouwen
0,49
Onbekend/geen
0,06
Onbekend/geen
0,06
Religie
Zoon
Zoon
Protestanten
0,59
-
-
Laag-/ongeschoolde arbeiders landbouw
0,15
Katholieken
0,33
Ongeschoolde arbeiders 0,27
Ongeschoolde arbeiders
0,17
Geen religie/onbekend
0,08
Laaggeschoolde arbeiders 0,15
Laaggeschoolde arbeiders
0,12
Regio van geboorte
-
-
Boeren en vissers
0,10
Noord-West
0,31
Geschoolde arbeiders
0,18
Voorlieden en geschoolde arbeiders
0,15
Zuid-Oost
0,27
Midden-/bovenklasse
0,24
Managers en professionals
0,17
West
0,42
Onbekend/geen
0,15
Onbekend/geen
0,14
Stedelijkheid geboortegemeente
Opwaartse mobiliteita
0,12
Opwaartse mobiliteita
0,17
Ruraal
0,61
Echtgenoot
Echtgenoot
Stedelijk
0,39
-
-
Laag-/ongeschoolde arbeiders landbouw
0,13
Ongeschoolde arbeiders 0,29
Ongeschoolde arbeiders
0,18
Laaggeschoolde arbeiders 0,13
Laaggeschoolde arbeiders
0,13
-
-
Boeren en vissers
0,12
Geschoolde arbeiders
0,23
Voorlieden en geschoolde arbeiders
0,19
Midden-/bovenklasse
0,32
Managers en professionals
0,22
Onbekend/geen
0,03
Onbekend/geen
0,02
Opwaartse mobiliteitb
0,18
Opwaartse mobiliteitb
0,21
a Ten opzichte van sociale status van de vader. b Ten opzichte van sociale status van de vader van de vrouw. Beschrijvende kenmerken van de studie Bronnen: HSN dataset ESM, release ESM.08 en HSN dataset Levenslopen, release 2007.01. 217 sociale klasse, sociale mobiliteit en sterfte in nederland Tabel 2 Proportie van de personen die de achttienjarige leeftijd hebben bereikt waarvoor geen informatie over de overlijdensdatum beschikbaar is,
per geboortecohorte, geslacht, geboorteregio, karakter geboortegemeente en sociale-klassenindeling (N=1871; 19 %)
Proportie
SOCPO-indeling
Proportie
HISCLASS-indeling
Proportie
Cohort 1850-1879
0,13
Vader
Vader
Cohort 1880-1899
0,15
-
-
Laag-/ongeschoolde arbeiders landbouw
0,17
Cohort 1900-1922
0,24
Ongeschoolde arbeiders
0,17
Ongeschoolde arbeiders
0,16
Laaggeschoolde arbeiders
0,19
Laaggeschoolde arbeiders
0,20
Mannen
0,17
-
-
Boeren en vissers
0,18
Vrouwen
0,20
Geschoolde arbeiders
0,20
Voorlieden en geschoolde arbeiders
0,20
Midden-/bovenklasse
0,20
Managers en professionals
0,22
Noord-west
0,17
Onbekend/geen
0,23
Onbekend/geen
0,23
Zuid-oost
0,20
West
0,20
Zoon
Zoon
-
Laag-/ongeschoolde arbeiders landbouw
0,13
Ruraal
0,18
Ongeschoolde arbeiders
0,13
Ongeschoolde arbeiders
0,13
Stedelijk
0,21
Laaggeschoolde arbeiders
0,15
Laaggeschoolde arbeiders
0,13
-
-
Boeren en vissers
0,12
Geschoolde arbeiders
0,14
Voorlieden en geschoolde arbeiders
0,15
Midden-/bovenklasse
0,13
Managers en professionals
0,14
Onbekend/geen
0,37
Onbekend/geen
0,40
Echtgenoot
Echtgenoot
-
-
Laag-/ongeschoolde arbeiders landbouw
0,09
Ongeschoolde arbeiders
0,08
Ongeschoolde arbeiders
0,08
Laaggeschoolde arbeiders
0,07
Laaggeschoolde arbeiders
0,15
-
-
Boeren en vissers
0,07
Geschoolde arbeiders
0,08
Voorlieden en geschoolde arbeiders
0,09
Midden-/bovenklasse
0,07
Managers en professionals
0,07
Onbekend/geen
0,16
Onbekend/geen
0,17
Bronnen: HSN dataset ESM, release ESM.08 en HSN dataset Levenslopen, release 2007.01 218 frans van poppel en ruben van gaalen tering van de overleving ten opzichte van de eerdere generaties. Ten opzichte
van de Nederlandse bevolking als geheel is sprake van een in de loop van de tijd
toenemende overschatting van de sterfte. Nog meer reliëf krijgt de overschatting
van de sterfte in het HSN-bestand wanneer men voor de verschillende cohorten
de levensverwachting van een achttienjarige vergelijkt met die van de Neder-
landse bevolking als geheel. In het oudste cohort vinden we voor de HSN-data
een waarde van 45,3 jaar, voor het identieke Nederlandse cohort een waarde van
48,4 jaar; voor het geboortecohort 1880-1899 loopt het verschil op tot 5,7 jaar (res-
pectievelijk 47,7 en 53,4 jaar) en in het oudste cohort bedraagt het verschil zelfs
6,4 jaar (50,5 tegenover 56,9 jaar). De conclusie moet zijn dat wat de temporele
ontwikkeling betreft de HSN-sterftedata niet representatief zijn voor Nederland
als geheel. We hebben echter geen duidelijke aanwijzingen dat dit ook eventuele
sociale-sterfteverschillen heeft beïnvloed. 219 Figuur 1 Overlevingscurve voor geboortecohorten, leeftijd 18 en hoger, HSN en nationale cijfers
Bronnen: niet-gepubliceerde generatiesterftetafels CBS (nationale cijfers), HSN-dataset ESM-release ESM.08
en HSN-dataset Levenslopen, release 2007.01. sociale klasse, sociale mobiliteit en sterfte in nederland 0
10
20
30
40
50
60
70
80
90
100
18
23
28
33
38
43
48
53
58
63
68
73
78
83
88
93
98
103
Leeftijd
Percentage nog in leven Bronnen: niet-gepubliceerde generatiesterftetafels CBS (nationale cijfers), HSN-dataset ESM-release ESM.08
en HSN-dataset Levenslopen, release 2007.01. Methode We schatten de sterftekans naar sociale klasse (volgens verschillende indelingen)
van de tussen 1850 en 1922 geborenen, vanaf het moment dat ze de leeftijd van
achttien hebben bereikt, gebruikmakend van gebeurtenissenanalyse (Blossfeld
& Rohwer, 2002; Cox, 1972). Een van de voordelen van deze methode, vergeleken
met lineaire regressie, is dat het ons in staat stelt rechts gecensureerde perso-
nen – personen van wie we slechts tot een bepaald moment informatie hebben
– in de analyse te betrekken. Omdat we voornamelijk zijn geïnteresseerd in de
verschillen in overlevingskansen tussen sociale klassen en niet in het effect van
leeftijd op zich, gebruiken we het Cox-model (Cox 1972). Dat model is een propor-
tional hazard-model en kan als volgt worden weergegeven: r(t) = h(t) exp (A(t)α). De
sterftekans, r(t), wordt benaderd door het product van een niet-gespecificeerde
basiskans, h(t), en een tweede term met mogelijke invloeden van een covariaat
vector A(t) op de sterftekans. 220 & Rohwer, 2002; Cox, 1972). Een van de voordelen van deze methode, vergeleken
met lineaire regressie, is dat het ons in staat stelt rechts gecensureerde perso-
nen – personen van wie we slechts tot een bepaald moment informatie hebben
– in de analyse te betrekken. Omdat we voornamelijk zijn geïnteresseerd in de
verschillen in overlevingskansen tussen sociale klassen en niet in het effect van
leeftijd op zich, gebruiken we het Cox-model (Cox 1972). Dat model is een propor-
tional hazard-model en kan als volgt worden weergegeven: r(t) = h(t) exp (A(t)α). De
sterftekans, r(t), wordt benaderd door het product van een niet-gespecificeerde
basiskans, h(t), en een tweede term met mogelijke invloeden van een covariaat
vector A(t) op de sterftekans. We richten ons op de overlevingskansen van die personen die ten minste
achttien jaar oud werden. We berekenen de sterfteleeftijd in dagen; was deze
niet bekend, dan is de datum van het vertrek uit het laatst bekende huishouden
als datum van laatste waarneming geselecteerd. We schatten vier verschillende
modellen. In het eerste model worden mannen en vrouwen samengenomen en
wordt alleen de sociale achtergrond opgenomen. In de drie andere modellen
wordt ook de eigen sociale positie geanalyseerd. Dit gebeurt apart voor mannen
(tabel 3) en gehuwde vrouwen (tabel 4). In het geval van de vrouwen wordt de
sociale klasse bepaald op basis van die van haar echtgenoot. Methode Van de 4.807 vrou-
wen is er voor 2.270 een huwelijksakte beschikbaar waarop het beroep van haar
echtgenoot wordt vermeld (de uitkomsten voor het eerste model zijn in beide
tabellen opgenomen). Alle modellen worden apart geschat voor de HISCLASS- en
de SOCPO-indeling. Resultaten Tabel 3 geeft de relatieve kansverhoudingen (hazard ratios) op sterfte na leeftijd
achttien weer. Het eerste model betreft het overlijden van mannen en vrouwen
gezamenlijk. De controlevariabelen laten een bekend beeld zien: de sterfte van
mannen is hoger (circa 30 procent) dan die van vrouwen, de sterfte in het eer-
ste cohort is duidelijk hoger dan die in het cohort 1880-1899, terwijl die weer
significant hoger ligt dan in het cohort 1900-1922. De in Noordwest-Nederland
geborenen hebben een lagere sterfte dan die in het westen en een nog lagere
dan die in het zuiden en oosten van Nederland. Gelet op de periode die we bestu-
deren (grofweg de jaren tachtig en volgende) is dat niet onverwacht, maar een
rol speelt ook dat er van de in Zuid- en Oost-Nederland geborenen alleen maar
relatief jong overledenen of niet-mobielen uit de geboorteperiode 1883-1922 in
de dataset zijn opgenomen, wat de sterfte enigszins opdrijft. Er worden geen significante verschillen in sterfte naar religie of stad/plat- frans van poppel en ruben van gaalen teland aangetroffen. De sterkte en de richting van de effecten van deze contro-
levariabelen verschillen overigens nauwelijks tussen de modellen die op de HIS-
CLASS- en de modellen die op de SOCPO-indeling zijn gebaseerd. 221 We zijn vooral geïnteresseerd in het effect van sociale klasse. In eerste instan-
tie bezien we daarvoor het effect van de sociale klasse van de vader op de sterfte
van de zonen en dochters. De uitkomsten blijken niet gevoelig te zijn voor de
gekozen indeling van sociale achtergrond: bij de HISCLASS-indeling wordt onder
de kinderen van boeren en vissers een significant lagere sterfte aangetroffen en
een niet-significant lagere bij de kinderen van managers en professionals. Deze
groepen komen min of meer overeen met de midden- en bovenklasse van de
SOCPO-indeling die ook een significant lager sterfterisico laat zien. Bovendien
zijn laag- en ongeschoolde arbeiders er in beide modellen niet echt slechter aan
toe. In tabel 3, model 2 (kolom 3 en kolom 7) wordt uitsluitend gekeken naar
de sociale verschillen in sterfte van de mannen, op basis van de eigen sociale
positie. Te constateren valt dat de effecten van de controlevariabelen slechts in
geringe mate veranderen. Resultaten Belangrijker is echter de constatering dat de eigen
sociale klasse van de man, gemeten op basis van het eerste beroep, geen statis-
tisch significant effect heeft op de sterfte; arbeiders in de landbouw hebben een
sterfterisico dat weinig afwijkt van dat van de voorlieden en geschoolde arbei-
ders, ongeschoolde arbeiders, managers of boeren. Een duidelijke gradiënt is
niet zichtbaar. Opvallend is hier het sterfteverlagend effect van het ontbreken
van een beroep dan wel de afwezigheid van informatie over de sociale klasse van
de zoon. Voor de SOCPO-indeling geldt een identiek verhaal. In model 3 (kolom 4 en kolom 8) wordt voor de beide klassenindelingen het
effect bezien zowel van de sociale klasse van oorsprong als van de eigen soci-
ale klasse. Opnieuw verandert er weinig aan de effecten van de controlevariabe-
len. De ouderlijke sociale klasse, hetzij volgens de HISCLASS-, hetzij volgens de
SOCPO-indeling, vertoont opnieuw weinig samenhang met het sterfteniveau van
volwassen mannen. Uitzondering vormen bij de SOCPO-indeling mannen die
stammen uit de sociale klasse van laaggeschoolde arbeiders; deze groep heeft
nu een significant verlaagd sterfteniveau. Verder is van een effect op het sterfte-
risico van de sociale klasse van de vader van de mannelijke onderzoekspersonen
geen sprake. De verschillen in het relatieve risico op overlijden op volwassen leef-
tijd van mannen die van oorsprong afkomstig zijn uit zo uiteenlopende sociale
klassen als bijvoorbeeld arbeiders in de landbouw of managers zijn te verwaarlo-
zen. Als gekeken wordt naar de sociale klasse die de betrokken mannen zelf heb-
ben bereikt, blijkt bijna zonder uitzondering van een effect van de eigen sociale
klasse geen sprake. Alleen mannen die zelf tot de laaggeschoolden behoren heb-
ben een statistisch significant hogere sterfte dan de referentiegroep. Opnieuw
blijkt dat de wijze waarop beroepen naar sociale klasse zijn ingedeeld weinig
effect heeft op de bevindingen. Resultaten (ref)
1,000
1,000
1,000
1,000
1,000
1,000
1,000
1,000
Geboorteregio
Noord-west
0,948
0,980
0,979
0,979
0,943*
0,977
0,969
0,969
Zuid-oost
1,066*
1,122**
1,127**
1,129**
1,064*
1,117*
1,116*
1,117*
West (ref)
1,000
1,000
1,000
1,000
1,000
1,000
1,000
1,000
Geboortegemeente
Ruraal
1,013
0,970
0,989
0,986
0,991
0,940
0,947
0,948
Stedelijk (ref)
1,000
1,000
1,000
1,000
1,000
1,000
1,000
1,000
HISCLASS vader
Laag-/ongeschoolde
arbeiders landbouw (ref.)
1,000
1,000
1,000
Ongeschoolde arbeiders
0,938
0,956
0,967
Laaggeschoolde arbeiders
0,953
0,967
1,104
Boeren en vissers
0,902*
0,914
0,948
Voorlieden en geschoolde
arbeiders
1,001
1,112
1,156* sociale klasse, sociale mobiliteit en sterfte in nederland
Managers en professionals
0,935
0,971
1,010
Onbekend/geen
0,982
0,975
1,011
HISCLASS zoon
Laag-/ongeschoolde
arbeiders landbouw (ref.)
1,000
1,000
1,000
Ongeschoolde arbeiders
0,937
0,961
0,972
Laaggeschoolde arbeiders
1,080
1,081
1,068
Boeren en vissers
0,876*
0,925
0,922
Voorlieden en geschoolde
arbeiders
1,000
0,969
0,960
Managers en professionals
1,024
1,024
1,018
Onbekend/geen
0,758***
0,763***
0,769***
SOCPO vader
Ongeschoold (ref.)
1,000
1,000
1,000
Laaggeschoold
0,925
0,879*
0,886*
Geschoolde arbeiders
1,020
1,063
1,085
Midden-/bovenklasse
0,944*
0,936
0,956
Onbekend/geen
0,994
0,950
0,967
SOCPO zoon
Ongeschoold (ref.)
1,000
1,000
1,000
Laaggeschoold
1,085
1,108*
1,101
Geschoolde arbeiders
1,031
1,018
1,002
Midden/bovenklasse
0,963
0,990
0,972
Onbekend/geen
0,821***
0,830**
0,826**
Mobiliteit
Neerwaarts gelijk (ref.)
1,000
1,000
Opwaarts
1,074
1,046
N
9826
5023
5023
5023
9826
5023
5023
5023
Events
7962
4153
4153
4153
7962
4153
4153
4153
Log-Likelihood
-65151,2
-31305,7
-31299,8
-31298,7
-65151,1
-31313,2
-31306,9
-31306,6
Nulmodel
-65483,8
-31407,1
-31407,1
-31407,1
-65483,8
-31407,1
-31407,1
-31407,1
* = p < 0,05; ** = p < 0,01; *** = p < 0,001. Bronnen: HSN dataset ESM release ESM 08 en HSN dataset Levenslopen release 2007 01 0,935
0,982 Resultaten Een laatste model (model 4) gaat expliciet in op de vraag of de sterfte van sociale klasse, sociale mobiliteit en sterfte in nederland sociale klasse, sociale mobiliteit en sterfte in nederland Cox-regressieanalyse van de tijdsduur tot overlijden vanaf leeftijd achttien (relatieve risico’s en significantieniveaus), mannen
HISCLASS-indeling
SOCPO-indeling
Model 1
Model 2
Model 3
Model 4
Model 1
Model 2
Model 3
Model 4
Zonen/
dochters HIS
CLASS vader
Zonen
HISCLASS
OP
Zonen
HISCLASS
Vader en OP
Zonen HIS
CLASS Vader
en OP/
mobiliteit
Zonen/
dochters
SOCPO
Vader
Zonen
SOCPO OP
Zonen
SOCPO
Vader en OP
Zonen
SOCPO Vader
en OP
en mobiliteit frans van poppel en ruben van gaalen
Tabel 3 Cox-regressieanalyse van de tijdsduur tot overlijden vanaf leeftijd achttien (relatieve risico’s en significantieniveaus), mannen
HISCLASS-indeling
SOCPO-indeling
Model 1
Model 2
Model 3
Model 4
Model 1
Model 2
Model 3
Model 4
Zonen/
dochters HIS
CLASS vader
Zonen
HISCLASS
OP
Zonen
HISCLASS
Vader en OP
Zonen HIS
CLASS Vader
en OP/
mobiliteit
Zonen/
dochters
SOCPO
Vader
Zonen
SOCPO OP
Zonen
SOCPO
Vader en OP
Zonen
SOCPO Vader
en OP
en mobiliteit
Geslacht
Mannen
1,277***
1,276***
Vrouwen (ref)
1,000
1,000
Geboorteperiode
1850-1879
1,278***
1,207***
1,211***
1,215***
1,280***
1,200***
1,210***
1,210***
1880-1899 (ref)
1,000
1,000
1,000
1,000
1,000
1,000
1,000
1,000
1900-1922
0,648***
0,753***
0,756***
0,759***
0,650***
0,747***
0,751***
0,751***
Religie
Protestanten
1,005
1,003
0,999
0,998
1,001
1,002
0,993
0,992
Katholieken
1,018
0,998
0,995
0,994
1,015
1,001
0,995
0,995
Geen religie/onb. sociale klasse, sociale mobiliteit en sterfte in nederland ox-regressieanalyse van de tijdsduur tot overlijden vanaf leeftijd achttien (relatieve risico’s en significantieniveaus), vrouwen
Hisclass-indeling
Socpo-indeling
Model 1
Model 2
Model 3
Model 4
Model 1
Model 2
Model 3
Model 4
Zonen/
dochters HIS
CLASS vader
Dochters
HISCLASS
echtgenoot
Dochters
HISCLASS
vader en
echtgenoot
Dochters
HISCLASS
vader en
echtgenoot
en mobiliteit
Zonen/
dochters
SOCPO vader
Dochters
SOCPO
echtgenoot
Dochters
SOCPO vader
en echtgenoot
Dochters
SOCPO vader
en echtgenoot
en mobiliteit Tabel 4 Cox-regressieanalyse van de tijdsduur tot overlijden vanaf leeftijd achttien (relatieve risico’s en significantieniveaus), vrouwen
Hisclass-indeling
Socpo-indeling
Model 1
Model 2
Model 3
Model 4
Model 1
Model 2
Model 3
Model 4
Zonen/
dochters HIS
CLASS vader
Dochters
HISCLASS
echtgenoot
Dochters
HISCLASS
vader en
echtgenoot
Dochters
HISCLASS
vader en
echtgenoot
en mobiliteit
Zonen/
dochters
SOCPO vader
Dochters
SOCPO
echtgenoot
Dochters
SOCPO vader
en echtgenoot
Dochters
SOCPO vader
en echtgenoot
en mobiliteit
Geslacht
Mannen
1,277***
1,276***
Vrouwen (ref)
1,000
1,000
Geboorteperiode
1850-1879
1,278***
1,376***
1,377***
1,371***
1,280***
1,378***
1,382***
1,381***
1880-1899 (ref)
1,000
1,000
1,000
1,000
1,000
1,000
1,000
1,000
1900-1922
0,648***
0,883*
0,879*
0,877*
0,650***
0,881*
0,878*
0,878*
Religie
Protestanten
1,005
0,989
0,985
0,980
1,001
0,997
0,994
0,993
Katholieken
1,018
0,938
0,933
0,927
1,015
0,943
0,941
0,941
Geen religie/onb. sociale klasse, sociale mobiliteit en sterfte in nederland (ref.)
1,000
1,000
1,000
1,000
1,000
1,000
1,000
1,000
Geboorteregio
Noord-west
0,948
0,845**
0,844**
0,850**
0,943*
0,849**
0,860**
0,860**
Zuid-oost
1,066*
0,947
0,952
0,953
1,064*
0,953
0,959
0,959
West (ref.)
1,000
1,000
1,000
1,000
1,000
1,000
1,000
1,000
Geboortegemeente
Ruraal
1,013
1,026
1,021
1,015
0,991
1,033
1,040
1,040
Stedelijk (ref.)
1,000
1,000
1,000
1,000
1,000
1,000
1,000
1,000
HISCLASS vader
Laag-/ongeschoolde
arbeiders landbouw (ref.)
1,000
1,000
1,000
Ongeschoolde arbeiders
0,938
1,004
1,014
Laaggeschoolde arbeiders
0,953
0,919
0,965
Boeren en vissers
0,902*
0,989
1,040
Voorlieden en geschoolde
arbeiders
1,001
1,039
1,092 Managers en professionals
0,935
0,960
1,008
Onbekend/geen
0,982
1,113
1,166
HISCLASS echtgenoot
Laag-/ongeschoolde
arbeiders landbouw (ref)
1,000
1,000
1,000
Ongeschoolde arbeiders
0,997
0,999
0,997
Laaggeschoolde arbeiders
0,981
0,981
0,971
Boeren en vissers
1,029
1,044
1,038
Voorlieden en geschoolde
arbeiders
0,883
0,887
0,880
Managers en professionals
0,991
1,005
0,992
Onbekend/geen
0,913
0,913
0,914
SOCPO vader
Ongeschoold (ref.)
1,000
1,000
1,000
Laaggeschoold
0,925
0,953
0,956
Geschoolde arbeiders
1,020
1,069
1,078
Midden-/bovenklasse
0,944*
1,021
1,029
Onbekend/geen
0,994
1,151
1,158
SOCPO echtgenoot
Ongeschoold (ref)
1,000
1,000
1,000
Laaggeschoold
1,076
1,078
1,077
Geschoolde arbeiders
0,935
0,936
0,931
Midden-/bovenklasse
1,022
1,021
1,014
Onbekend/geen
0,985
0,979
0,977
Mobiliteit
Neerwaarts gelijk (ref.)
1,088
1,000
Opwaarts
1,089
1,015
N
9826
2270
2270
2270
9826
2270
2270
2270
Events
7962
2092
2092
2092
7962
2092
2092
2092
Log-Likelihood
-65151,2
-14022,0
-14022,0
-14019,2
-65151,1
-14022,6
-14020,6
-14020,6
Nulmodel
-65483,8
-14053,7
-14053,7
-14053,7
-65483,8
-14053,7
-14053,7
-14053,7
* = p < 0,05; ** = p < 0,01; *** = p < 0,001. Bronnen: HSN dataset ESM, release ESM.08 en HSN dataset Levenslopen, release 2007.01. sociale klasse, sociale mobiliteit en sterfte in nederland sociaal mobiele mannen afwijkt van die van mannen die in dezelfde sociale klas-
se bleven als hun vader, dan wel daalden op de sociale ladder. Welke indeling
ook wordt aangehouden, een effect van sociale mobiliteit op sterfte wordt niet
aangetroffen. 226 Voor vrouwelijke onderzoekspersonen zijn eveneens afzonderlijke modellen
geschat. Deze betreffen echter alleen gehuwde vrouwen, en de ‘eigen’ sociale
klasse van deze vrouwen is bepaald aan de hand van de sociale klasse waartoe
haar echtgenoot behoort. De betreffende gegevens zijn vermeld in tabel 4. De uit-
komsten voor model 1 (kolom 2 en 6) zijn identiek aan die in tabel 3. Kolom 3 en
kolom 7 (model 2) geven de effecten weer van de sociale klasse van de echtgenoot
op de sterfte van hun vrouw. Bij vrouwen is noch bij de HISCLASS-classificatie, noch bij de SOCPO-indeling
sprake van een duidelijke gradiënt in sterfte naar sociale klasse van de echtge-
noot, min of meer in lijn met hedendaags onderzoek waaruit blijkt dat de socia-
le-klasseverschillen bij vrouwen bijna zonder uitzondering geringer zijn dan bij
mannen (Mustard & Etches, 2003). Model 3 (kolom 4 en kolom 8) combineert de
effecten van de sociale klasse van oorsprong en die van de ‘eigen’ sociale klasse. Opnieuw is van statistisch significante effecten op het sterfteniveau van de soci-
ale klasse van oorsprong of van die van de sociale klasse waartoe men door het
huwelijk is gaan behoren geen sprake. In het laatste model is ook bij de vrouwen
naar de effecten van intergenerationele sociale mobiliteit gekeken; ook bij de
volwassen vrouwen blijkt dat proces geen effect te hebben gehad op de sterfteri-
sico’s. Al met al constateren we dat zowel in de HISCLASS-indeling als volgens de
SOCPO-indeling zelden of nooit van statistisch significante verschillen in sterfte
sprake is, en dat een duidelijke sociale gradiënt ver te zoeken is.9 Discussie De centrale vraag in dit hoofdstuk was of er in de tweede helft van de negen-
tiende en in de twintigste eeuw bij volwassenen verschillen in sterfte naar
sociale klasse worden gevonden en zo ja, hoe deze verschillen zich over de tijd
heen hebben ontwikkeld. We gingen allereerst na of de sociale klasse waaruit
een persoon afkomstig was (gemeten via het beroep van de vader, volgens een
tweetal klassenindelingen) nog effect had op de sterfte op volwassen leeftijd
(na leeftijd 18). Een dergelijk effect werd zelden aangetroffen, het meest con-
sequent nog bij kinderen van boeren die een lager sterfterisico kenden. Vervol-
gens keken we voor mannen naar het effect van de sociale klasse van de per-
soon zelf, gemeten op basis van het eerste bereikte beroep. Ook hier vonden we
voor beide klassenindelingen eenzelfde resultaat, namelijk de bijna complete
afwezigheid van samenhang tussen eigen sociale klasse en sterfte op volwassen
leeftijd en in elk geval geen duidelijke gradiënt. Tot slot is specifiek voor vrou-
wen gekeken naar de samenhang tussen sterfte enerzijds, de sociale klasse van frans van poppel en ruben van gaalen de vader en die van de echtgenoot van de vrouw anderzijds. Ook hier werd geen
aanwijzing gevonden voor een effect van de sociale klasse van oorsprong of van
de ‘eigen’ sociale klasse op de sterfte. Belangrijk is de constatering dat het klas-
senschema dat is gehanteerd op de sterfteverschillen naar sociale klasse geen
effect heeft. 227 Vergeleken met de sterfteverschillen naar sociaal-economische status die
tegenwoordig in Nederland worden gevonden (de levensverwachting bij de
geboorte verschilt 4,8-5,0 jaar naar sociale klasse, zie Perenboom, Van Herten,
Boshuizen & Van den Bos, 2005) zijn de door ons gevonden cijfers uitzonderlijk
te noemen. De constatering dat sociale klassenverschillen in sterfte van volwas-
senen in onze historische cohorten afwezig zijn is ook in strijd met het over-
geleverde historiografische beeld. Dat betekent echter niet dat deze bevinding
onjuist hoeft te zijn. De stelling van Antonovsky dat na 1850 het gat in de levens-
verwachting tussen de sociale klassen afnam is bijvoorbeeld niet in strijd met
onze observaties. Er zijn ook andere historische studies die vergelijkbare uitkom-
sten vonden. Bourdieu en Kesztenbaum (2004) bijvoorbeeld constateerden in
Frankrijk in de geboortecohorten van 1810 tot 1850 betrekkelijk kleine verschil-
len tussen rijk en arm in de levensverwachting op dertigjarige leeftijd, in de orde
van grootte van 1,5 tot 2,2 jaar. Discussie Razzell en Spence (2006) en Smith (1983) stelden
voor respectievelijk Engeland en de VS vast dat er vóór de twintigste eeuw geen
sprake was van een samenhang tussen de sociaal-economische status en de sterf-
te op volwassen leeftijd. Onze bevindingen vinden ook steun in studies waarin,
op basis van macrodata betrekking hebbend op lengtegegevens en sterftecijfers,
werd aangetoond dat een verslechtering van de gezondheidssituatie geen onver-
mijdelijke consequentie was van de negentiende-eeuwse processen van industri-
alisatie en verstedelijking (Sandberg & Steckel, 1997; Weir, 1997). De vraag is echter wel hoe hard onze bevindingen zijn. We hebben moe-
ten constateren dat de studie van trends in sociale-klasseverschillen in sterfte
op basis van de huidige, nog niet complete HSN-dataset niet probleemloos is. Hoewel de uit de HSN af te leiden sterftematen over het algemeen qua richting
wel overeenkomen met die welke uit nationale statistieken voor de betreffende
periode bekend zijn (bijvoorbeeld wat betreft de regionale en sekseverschillen),
is vooral de verandering in de tijd in strijd met wat we uit die nationale statis-
tieken kunnen afleiden. De HSN-gegevens overschatten in sterke mate de sterfte,
en doen dat meer naarmate het om recentere cohorten gaat. Om dat effect te
reduceren is intensivering van de verzameling van nu nog ontbrekende sterfte-
data van onderzoekspersonen en/of koppeling van HSN-gegevens aan het Sociaal-
Statistisch Bestand van het CBS om te achterhalen welke onderzoekspersonen op
dit moment nog in leven zijn, noodzakelijk. Daarnaast is het zinvol om gebruik
te maken van recent ontwikkelde statistische methoden om het effect te bestu-
deren van de gevonden samenhang in de HSN-data tussen het bestudeerde sterf-
teproces en de met de dataverzameling samenhangende censurering (zie bijvoor-
beeld: Siannis, Copas & Lu, 2005). sociale klasse, sociale mobiliteit en sterfte in nederland Op voorhand hebben we geen reden om te stellen dat het ontbreken van de
overlijdensdatum vaker voorkomt in bepaalde sociale klassen, en daardoor een
verklaring zou kunnen bieden voor de afwezigheid van uitgesproken klassen-
verschillen. Er staan meerdere wegen open om na te gaan in hoeverre de afwe-
zigheid van klassenverschillen een artefact is van de wijze waarop de data zijn
geanalyseerd. De aandacht zou niet op het eerste maar op het hoogste uitgeoe-
fende beroep gericht kunnen worden. We zouden ook kunnen analyseren of er
inderdaad sprake is van een proportionaliteit van de effecten van sociale klasse. Discussie De survival-lijnen van de verschillende cohorten in figuur 1 laten zien dat van
volledige proportionaliteit geen sprake is. Daarbij moet wel worden opgemerkt
dat Cox-modellen over het algemeen wel robuuste schattingen leveren, ook bij
afwijkingen van proportionaliteit. Er zijn aanwijzingen dat op jongere leeftijd er
wel sprake is van een duidelijke samenhang tussen sociale klasse en sterfte. Dit
zou onderzocht kunnen worden door afzonderlijke modellen voor leeftijdsgroe-
pen te schatten. Selectieve sterfte verklaart wellicht de afwezigheid van verschil-
len op volwassen leeftijden. Een dergelijk effect valt te schatten met behulp van
sample selection-modellen (zie bijvoorbeeld Winship & Mare, 1992). 228 Voorlopig echter is de conclusie dat de standaardopvattingen over de desas-
treuze effecten van de negentiende- en begin-twintigste-eeuwse processen van
industrialisatie en verstedelijking op de gezondheid en de levensduur van de
arbeidende klasse niet voor Nederland lijken op te gaan. We vonden in elk geval
geen aanwijzing dat de situatie van de arbeidende klasse – of het nu gaat om die
in de landbouw of in de industrie – in deze periode sterk en ongunstig afweek
van die van de midden- en hogere sociale lagen. Het is mogelijk dat in enkele spe-
ciale sectoren van de industrie de sterfte hoog was en de levensverwachting lager
lag die van personen op midden of hogere posities, maar in zijn algemeenheid
gold dat zeker niet voor de arbeidende klasse. Het is zeer wel mogelijk dat dit
veroorzaakt is door de specifieke situatie van Nederland: hier was immers sprake
van relatief hoge lonen en een naar verhouding genereus systeem van armen-
zorg, die er voor zorgden dat de levensstandaard niet al te zeer verslechterde. In
Nederland was ook, anders dan in bijvoorbeeld Engeland, Frankrijk of Duitsland,
in de tweede helft van de negentiende eeuw geen sprake van het ontstaan van
een omvangrijk klassiek proletariaat dat opeengehoopt leefde in grootstedelij-
ke krottenwijken. Maar ook het agrarische proletariaat dat te kampen had met
een langdurige crisis in de landbouw in de jaren 1880 en 1890 werd niet gecon-
fronteerd met hogere sterfterisico’s (Wintle, 2000). De economische situatie in
Nederland verbeterde na 1864 continu (Van Zanden & Van Riel, 2004), en een
gezondheidsbeleid dat gericht was op verbetering van de volksgezondheid en
vermindering van de gezondheidsverschillen kwam na 1875 van de grond, deels
gestimuleerd door dezelfde economische groei (Mackenbach, 1992). Noten 1. We beperken ons hier bewust tot sterfteverschillen op volwassen leeftijd. Aan sociale ver-
schillen in sterfte onder kinderen is relatief meer aandacht besteed, ook in Nederland
(Van Poppel, Jonker & Mandemakers, 2005), maar duidelijk is wel dat het effect van SES
(maar ook van omgevingsfactoren) sterk verschilt per leeftijd, zeker in historische popula-
ties (Currie & Stabile, 2002; Ferrie, 2003; Garrett, Reid, Schürer & Szreter, 2001). Over het
algemeen reageerde de sterfte onder kinderen minder op de variatie in de sociale positie
dan die van volwassenen, omdat de mate waarin borstvoeding werd gegeven en de leeftijd
waarop deze praktijk stopte van meer belang waren voor de overlevingskansen van het
kind dan strikt economische factoren als de woonsituatie en kwaliteit van de voeding;
deze praktijken waren minder afhankelijk van de sociaal-economische positie. p
j
j
p
2. Een enigszins vergelijkbare opvatting over het relatieve belang van sociale klasse versus
ruimtelijke verschillen wordt ingenomen door Smith (1983). In afwijking van Woods c.s. meent hij echter dat vóór 1880 de sterftekansen van de verschillende sociale klassen niet
veel van elkaar afweken. Pas op het moment dat eind negentiende eeuw de meer welva-
rende groepen zich ruimtelijk distantieerden van de door hoge sterfte gekenmerkte gebie-
den namen de verschillen tussen de sociale klassen toe. Het blijft echter een niet verder
getoetste hypothese. 3. Op basis van provinciale sterftetafels voor de periode 1850-1859, gebaseerd op eigen bere-
keningen. Deze verschillen werden vooral veroorzaakt door uiteenlopende zuigelingen-
en kindersterftecijfers, maar ook de levensverwachting van een vijfjarige in Zeeland liep
nog circa zes jaar achter op die van een leeftijdsgenoot uit Noord- of Oost-Nederland 4. Onze dank gaat uit naar Dr. Andrew Miles (University of Birmingham) en Dr. Bart van
de Putte (Universiteit Leuven) voor hun hulp bij de codering van de beroepstitels in het
SOCPO-schema. Dr. Marco van Leeuwen (Internationaal Instituut voor Sociale Geschiede-
nis, Amsterdam) stelde een elektronisch bestand met een groot aantal HISCO-codes van
beroepen ter beschikking. 5. Voor HISCLASS zijn alleen die personen gedefinieerd als opwaarts mobiel (1) wiens vader
de HISCLASS-code 12 had, terwijl zijzelf of hun echtgenoot HISCLASS-code 10 of lager had-
den of (2) wiens vader de HISCLASS-code 11 had, terwijl zijzelf of hun echtgenoot HIS-
CLASS-code 9 of lager hadden. Alleen de van origine ongeschoolden die zich minstens 5. Discussie Ook voor
andere landen is geconstateerd dat gedurende de negentiende en vroeg-twintig-
ste eeuw in het bijzonder de lagere sociale klassen profijt hadden van de nieuwe frans van poppel en ruben van gaalen mogelijkheden die gecreëerd werden door de toegenomen medische kennis, ver-
beterde hygiëne en de met de economische groei samenhangende verbeteringen
in de voedingstoestand, de aanleg van rioleringen, waterleiding, en bijvoorbeeld
de verbeterde woonsituatie (Ferrie, 2003; Rogers Hollingsworth, 1981). 229 Een tweede conclusie is een bevestiging van wat eerder buitenlands onder-
zoek ons heeft geleerd, namelijk dat waar men leefde van grotere betekenis was
voor de sterfterisico’s dan tot welke sociale klasse men behoorde (Chaunu, 1972). Dat gold ook voor het negentiende-eeuwse Nederland. Helaas kon met die regi-
onale factor slechts in beperkte mate rekening worden gehouden. In principe
zou door een meer gedifferentieerde regionale indeling en de introductie in de
analyse van sterfte-indicatoren op het niveau van de gemeente waar de onder-
zoekspersonen woonachtig waren, op genuanceerdere wijze deze context in de
analyse kunnen worden betrokken. Ook de introductie van interactie-effecten
(regio en sociale klasse) zou een oplossing kunnen bieden. Noten We hebben ook aparte modellen geschat voor de sterftekansen tussen de leeftijd 15 en 25 en
vonden daarbij effecten van de hoogte van de eigen sociale status op de sterftekansen in die
leeftijdsgroep. Noten Voor HISCLASS zijn alleen die personen gedefinieerd als opwaarts mobiel (1) wiens vader
de HISCLASS-code 12 had, terwijl zijzelf of hun echtgenoot HISCLASS-code 10 of lager had-
den of (2) wiens vader de HISCLASS-code 11 had, terwijl zijzelf of hun echtgenoot HIS-
CLASS-code 9 of lager hadden. Alleen de van origine ongeschoolden die zich minstens sociale klasse, sociale mobiliteit en sterfte in nederland twee stappen hebben verbeterd zijn dus als mobiel beschouwd. Voor SOCPO zijn alleen die
personen gedefinieerd als opwaarts mobiel (1) wiens vader de SOCPO-code 1 had, terwijl
zijzelf of hun echtgenoot SOCPO-code 3 of hoger hadden of (2) wiens vader de SOCPO-code
2 had, terwijl zijzelf of hun echtgenoot SOCPO-code 4 of hoger hadden. Alleen de van ori-
gine ongeschoolden of laaggeschoolden, die zich minstens twee stappen verbeterden zijn
dus als opwaarts mobiel (1) beschouwd. twee stappen hebben verbeterd zijn dus als mobiel beschouwd. Voor SOCPO zijn alleen die
personen gedefinieerd als opwaarts mobiel (1) wiens vader de SOCPO-code 1 had, terwijl
zijzelf of hun echtgenoot SOCPO-code 3 of hoger hadden of (2) wiens vader de SOCPO-code
2 had, terwijl zijzelf of hun echtgenoot SOCPO-code 4 of hoger hadden. Alleen de van ori-
gine ongeschoolden of laaggeschoolden, die zich minstens twee stappen verbeterden zijn
dus als opwaarts mobiel (1) beschouwd. 230 6. De gemiddelde overlijdensleeftijd van de na het achttiende levensjaar overleden personen
bedroeg 67,9 jaar. Van hen voor wie geen overlijdensdatum is gevonden was de leeftijd bij
laatste waarneming 46,7 jaar. g
j
7. De aanname dat alle nog niet gevonden onderzoekspersonen uit de recentere cohorten
inderdaad nog in leven zijn leidt tot een lichte onderschatting van het sterfteniveau. 7. De aanname dat alle nog niet gevonden onderzoekspersonen uit de recentere cohorten
inderdaad nog in leven zijn leidt tot een lichte onderschatting van het sterfteniveau. g
j
g
8. Voor de HSN-data zijn sterftetafels berekend die zijn vergeleken met ongepubliceerds CBS-
generatiesterftetafels. De CBS-data zijn ongewogen gemiddelden van mannen- en vrouwens-
terftetafels. 8. Voor de HSN-data zijn sterftetafels berekend die zijn vergeleken met ongepubliceerds CBS-
generatiesterftetafels. De CBS-data zijn ongewogen gemiddelden van mannen- en vrouwens-
terftetafels. 9. We hebben ook aparte modellen geschat voor de sterftekansen tussen de leeftijd 15 en 25 en
vonden daarbij effecten van de hoogte van de eigen sociale status op de sterftekansen in die
leeftijdsgroep. 9. Literatuur Antonovsky, A. (1967). Social class, life expectancy and overall mortality. Milbank
Memorial Fund Quarterly, 45, 31-76. Antonovsky, A. (1967). Social class, life expectancy and overall mortality. Milbank
Memorial Fund Quarterly, 45, 31-76. Backs, J. (2001). Mortality in Ghent, 1850-1950. A social analysis of death. Revue Belge
d’histoire contemporaine, 31, 529-556. Bengtsson, T. (2004). Mortality and social class in four Scanian parishes, 1766-1865. In T. Bengtsson, C. Campbell, J.Z. Lee e.a. (red.), Life under pressure. Mortality and
living standards in Europe and Asia, 1700-1900 (pp. 135-171). Cambridge, Ma: The
MIT Press. Bie, R. de (2006). Gezonde en ongezonde beroepen. Bevolkingstrends, 54(1), 10-11. Blossfeld, H.-P. & G. Rohwer (2002). Techniques of event history modelling. New approa-
ches to causal analysis. Mahwah: NJ.: Erlbaum. Blum, A., J. Houdaille. & M. Lamouche (1990). Mortality differentials in France
during the late 18th and early 19th centuries. Population. English Selection, 2, 163-
185. Borrell, C., E. Regidor, L.C. Arias., P. Navarro, R. Puigpinos, V. Dominguez & A. Plasen-
cia (1999). Inequalities in mortality according to educational level in two large
Southern European cities. International Journal of Epidemiology, 28(1), 58-63. Bourdieu, J. & L. Kesztenbaum (2004). Vieux, riches et bien portants. Une applicati-
on de la base «TRA» aux liens entre mortalité et richesse. Annales de démographie
historique, 1, 79-105. Breschi, M., R. Derosas & M. Manfredini (2004). Mortality and environment in three
Emilian, Tuscan, and Venetian communities, 1800-1883. In T. Bengtsson, C. Campbell, J.Z. Lee e.a. (red.), Life under pressure. Mortality and living standards in
Europe and Asia, 1700-1900 (pp. 209-252). Cambridge, Ma: The MIT Press. Broes van Dort, K. (1861). Bijdrage tot de kennis van de sterfte der gemeente Goes en van frans van poppel en ruben van gaalen den gemiddelden en waarschijnlijken levensduur harer inwoners gedurende het 30 jarige
tijdvak (1830 1859). Goes: L. de Fouw. den gemiddelden en waarschijnlijken levensduur harer inwoners gedurende het 30 jarige
tijdvak (1830 1859). Goes: L. de Fouw. Brugmans, I.J. (1975). De arbeidende klasse in Nederland in de 19e eeuw 1813-1870. Utrecht: Spectrum. Brugmans, I.J. (1975). De arbeidende klasse in Nederland in de 19e eeuw 1813-1870. Utrecht: Spectrum. 231 Buchner, E.C. (1852). Bijdragen tot de statistiek der sterfte in de gemeente Amsterdam
gedurende de laatste 12 jaren. Amsterdam: s.n. Büchner, W.F. (1842). Bijdragen tot de geneeskundige topographie en statistiek van
Gouda. Gouda: Van Goor. Cambois, E. (2004). Literatuur Careers and mortality in France: evidence on how far occupa-
tional mobility predicts differentiated risks. Social Science & Medicine, 58, 2545-
2558. Centraal Bureau voor de Statistiek (1906). Statistiek van de sterfte onder de mannen,
met onderscheiding naar het beroep en in verband met leeftijden en doodsoorzaken in
de jaren 1896-1900. ’s-Gravenhage: Belinfante. Centraal Bureau voor de Statistiek. (1912). Statistiek van de sterfte onder de mannen,
met onderscheiding naar het beroep en in verband met leeftijden en doodsoorzaken in
de jaren 1896-1903. ’s-Gravenhage: Belinfante. Centraal Bureau voor de Statistiek. (1917). Statistiek van de sterfte onder de mannen
van 18-65 jaar met onderscheiding naar beroep en de positie daarin bekleed in verband
met leeft den en doodsoorzaken in de jaren 1908-1911. ’s-Gravenhage: Belinfante. Centrale Commissie voor de Statistiek. (1898). Statistiek der sterfte in de jaren 1891-
1895 van mannen van 18 tot en met 50 jaar, met onderscheiding naar het beroep, den
leeft d en de doodsoorzaken. ’s-Gravenhage: Belinfante. Chaunu, P. (1972). Malthusianisme démographique et Malthusianisme écono-
mique. Réflexions sur l’échec de la Normandie à l’époque du Démarrage. Annales E.S.C., 27(1), 1-19. Coronel, S.S. (1861). De gezondheidsleer toegepast op de fabrieksnijverheid. Een handboek
voor industriëlen, genees- en staathuishoudkundigen. Haarlem: De Erven Loosjes. Coronel, S.S. (1862). De bevolking van Hilversum in verband tot hare industrie;
eene statistische studie. Nederlandsch Tijdschrift voor Geneeskunde, 6, 651-664. Coronel, S.S. (1864a). De diamantwerkers te Amsterdam. Eene hygiënische stu-
die. Nederlandsch Tijdschrift voor Geneeskunde, 8, 633-650. Coronel, S.S. (1864b). De Leidsche wolfabrieken en haar invloed op de gezond-
heid der arbeiders. Nederlandsch Tijdschrift voor Geneeskunde, 8, 225-239. Cox, D. (1972). Regression models and life tables (with discussion). Journal of the
Royal Statistical Society Series B, 34, 187-220. Craig, P. & J. Forbes (2005). Social position and health: Are old and new occupatio-
nal classifications interchangeable? Journal of Biosocial Science, 37, 89-106. Currie, J. & M. Stabile (2002). Socioeconomic status and health: why is the relationship
stronger for older children? (Working paper 9098). Cambridge, Ma: National
Bureau of Economic Research. Davey Smith, G., D. Blane & M. Bartley (1994). Explanations for socio-economic
differentials in mortality. European Journal of Public Health, 4, 131-144. sociale klasse, sociale mobiliteit en sterfte in nederland Davey Smith, G., D. Dorling & M. Shaw (2001). Poverty, inequality and health in Bri-
tain. 1800-2000: A reader. Bristol: The Policy Press. Davey Smith, G., C. Hart, D. Hole, P. MacKinnon, C. Gillis, G. Watt, D. Literatuur Blane &
V. Hawthorne (1998). Education and occupational social class: which is the
more important indicator of mortality risk? Journal of epidemiology and com-
munity health, 52, 153-160. 232 Engels, F. (1976). De toestand van de arbeidersklasse in Engeland. Naar eigen aanschou-
wingen en authentieke bronnen. Moskou: Uitgeverij Progres. Ferrie, J.P. (2003). The rich and the dead: socioeconomic status and mortality in
the United States, 1850-60. In D.L. Costa (red.), Health and labor force participa-
tion over the life cycle. Evidence from the past (pp. 11-50). Chicago: The University
of Chicago Press. Garrett, E., A. Reid, K. Schürer & S. Szreter (2001). Changing family size in England
and Wales. Place, class and demography, 1891 1911. Cambridge: Cambridge Uni-
versity Press. Giele, J.J. & G.J. Van Oenen (1974). De sociale struktuur van de Nederlandse
samenleving rond 1850. Mededelingen van de Nederlandse Vereniging voor Sociale
Geschiedenis, 45, 2-32. Harding, S. (1995). Social class differences in mortality of men: recent evidence
from the OPCS Longitudinal Study. Office of Population Censuses and Sur-
veys. Popul Trends, 80, 31-37. Harris, B. (1999). Morbidity and mortality during the health transition: a com-
ment on James C. Riley, ‘Why sickness and death rates do not move parallel to
one another over time’. Social History of Medicine, 12, 125-131. Hengel, J.F. van (1875). Geneeskundige plaatsbeschrijving van het Gooiland. ’s-Graven-
hage: Van Weelden en Mingelen. Houwaart, E.S. (1991). De hygiënisten. Artsen, staat en volksgezondheid in Nederland
1840-1890. Groningen: Historische Uitgeverij Groningen. Hummer, R.A., R.G. Rogers, C.B. Nam & C.G. Ellison (1999). Religious involvement
and U.S. adult mortality. Demography, 36, 273-285. Johansson, S.R. & A.B. Kasakoff (2000). Mortality history and the misleading
mean. Historical methods, 33, 56-58. Joung, I.M.A., J.J. Glerum, F.W.A. van Poppel, J.W.P.F. Kardaun & J.P. Mackenbach
(1996). The contribution of specific causes of death to mortality differences by
marital status in the Netherlands. European Journal of Public Health, 6, 142-149. Krieger, N. & E. Fee (1996). Measuring social inequalities in health in the United
States: a historical review, 1900-1950. International Journal of Health Services, 26,
391-418. Krieger, N., D.R. Williams & N.E. Moss (1997). Measuring social class in US public
health research: concepts, methodologies, and guidelines. Annual Review of
Public Health, 18, 341-378. Kunst, A.E. (1997). Cross-national comparisons of socio-economic differences in mortality. Rotterdam: Erasmus University. frans van poppel en ruben van gaalen Kunst, A.E., V. Bos, P. Santana, T. Valkonen, J.P. Mackenbach, O. Andersen, M. Literatuur Car-
dano, G. Costa, S. Harding, O. Hemström, R. Layte, E. Regidor & A. Reid (2004). Monitoring of trends in socioeconomic inequalities in mortality. Experiences
from a European project. Demographic Research, Special collection 2, Determinants
of diverging trends in mortality, 2004 (April), 229-254. 233 Leeuwen, M.H.D. van & I. Maas (2005). HISCLASS: A Historical International Social
Class Scheme for occupational titles in the past. Unpublished manuscript. Leeuwen, M.H.D. van, I. Maas. & A. Miles (2002). HISCO: Historical International
Standard Classification of Occupations. Leuven: Leuven University Press. Levin, J.S. (1994). Religion and health: is there an association, is it valid, and is it
causal? Social Science & Medicine, 38, 1475-1482. Leye, R. & D. Joye (1994). What is Switzerland’s stratification like: classes, prestige
gradation, professional categories? International Sociology, 9, 313-336. Link, B.G., M.E. Northridge, J.C. Phelan M.L. & Ganz (1998). Social epidemiology
and the fundamental cause concept: on the structuring of effective cancer
screens by socioeconomic status. Milbank Quarterly, 76, 375-402. Link, B.G. & J.C. Phelan (1995). Social conditions as fundamental causes of disease. Journal of Health and Social Behavior, 35, 80-94. Link, B.G. & J.C. Phelan (1996). Understanding sociodemographic differences in
health-the role of fundamental social causes. American Journal of Public Health,
86, 471-473. Link, B.G. & J.C. Phelan (2002). McKeown and the idea that social conditions are
fundamental causes of disease. American Journal of Public Health, 92, 730-732. Lundberg, O. (1991). Childhood living conditions, health status and social mobi-
lity: a contribution to the health selection debate. European Sociological Review,
7, 149-162. Lundberg, O. (1993). The impact of childhood living conditions on illness and
mortality in adulthood. Social Science & Medicine, 36, 1047-1052. Lutfey, K. & J. Freese (2005). Toward some fundamentals of fundamental causa-
lity: socioeconomic status and health in the routine clinic visit for diabetes. American Journal of Sociology, 110, 1326-1372. Mackenbach, J.P. (1992). De veren van Icarus: over de achtergronden van twee eeuwen
epidemiologische transities in Nederland. Utrecht: Wetenschappelijke Uitgeverij
Bunge. Mackenbach, J.P. (1994). Ongezonde verschillen. Over sociale stratificatie en gezondheid
in Nederland. Assen: Van Gorcum. Mackenbach, J.P., V. Bos, O. Andersen, M. Cardano, G. Costa, S. Harding, A. Reid,
O. Hemstrom, T. Valkonen & A.E. Kunst (2003). Widening socioeconomic ine-
qualities in mortality in six Western European countries. International Journal
of Epidemiology, 32, 830-837. Mandemakers, K. (2000). The Netherlands. Historical Sample of the Netherlands. In P. Kelly Hall, R. McCaa & G. Literatuur Thorvaldsen (red.), Handbook of International His-
torical Microdata for Population Research (pp. 149-177). Minneapolis: Minnesota
Population Center. sociale klasse, sociale mobiliteit en sterfte in nederland sociale klasse, sociale mobiliteit en sterfte in nederland Mandemakers, K. (2001). Historical sample of the Netherlands HSN. Historical
Social Research, 26(4), 179-190. Manor, O., S. Matthews & C. Power (2003). Health selection: the role of inter- and
intra-generational mobility on social inequalities in health. Social Science &
Medicine, 57, 2217-2227. 234 Mare, R.D. (1990). Socio-economic careers and differential mortality among older
men in the United States. In J. Vallin, S. D’Souza & A. Palloni (red.), Measu-
rement and analysis of mortality. New Approaches (pp. 362-387). Oxford: Oxford
Clarendon Press. Marmot, M. (2004). The status syndrome: how social standing affects our health and lon-
gevity. London: Bloomsbury Publ. Martikainen, P., J. Blomgren & T. Valkonen (2007). Change in the total and inde-
pendent effects of education and occupational social class on mortality: ana-
lyses of all Finnish men and women in the period 1971-2000. Journal of Epide-
miology and Community Health, 61, 499-505. Mustard, C.A. & J. Etches (2003). Gender differences in socioeconomic inequality
in mortality. Journal of Epidemiology and Community Health, 57, 974-980. Neven, M. (2000). Mortality differentials and the peculiarities of mortality in an
urban-industrial population: a case study of Tilleur, Belgium. Continuity and
Change, 15, 297-329. Oris, M. (1998). Mortalité, industrialisation et urbanisation au XIXe siècle. Quelques résultats des recherches Liégoises. In C. Desama & M. Oris (red.), Dix
essais sur la démographie urbaine de la Wallonie au XIXe siècle (pp. 289-321). Bruxel-
les: Crédit Communal. Pamuk, E.R. (1985). Social class inequality in mortality from 1921 to 1972 in
England and Wales. Population Studies, 39, 17-31. Perenboom, R.J., L.M. van Herten, H.C. Boshuizen & G.A. van den Bos (2005). Life
expectancy without chronic morbidity: trends in gender and socioeconomic
disparities. Public Health Review, 46-54. Phelan, J.C., B.G. Link, A. Diez-Roux, I. Kawachi & B. Levin (2004). “Fundamental
causes” of social inequalities in mortality: a test of the theory. Journal of Health
and Social Behavior, 45, 265-285. Poppel, F. van (1989). Urban-rural versus regional differences in demographic
behavior. The Netherlands, 1850-1960. Journal of Urban History, 15, 363-398. Poppel, F. van & E. Beekink (2003). De ‘gezondheid’ van Nederland. Sterftetrends
en sterfteverschillen in de negentiende en twintigste eeuw. In E. Beekink, O. Boonstra, T. Engelen & H. Knippenberg (red.), Nederland in verandering. Literatuur Maat-
schappelijke ontwikkelingen in kaart gebracht 1800-2000 (pp. 71-94). Amsterdam:
Aksant. Poppel, F. van, I. Deerenberg, J. Wolleswinkel-van den Bosch & P. Ekamper (2005). Hoe lang leefden wij? Veranderingen in de levensduur en het doodsoorzaken-
patroon. Bevolkingstrends, 53, 13-25. Poppel, F. van, M. Jonker & K. Mandemakers (2005). Differential infant and child frans van poppel en ruben van gaalen mortality in three Dutch regions, 1812-1909. Economic History Review, 58, 272-
309. Poppel, F. van & I. Joung (2001). Long-term trends in marital status mortality dif-
ferences in The Netherlands 1850-1970. Journal of Biosocial Science, 33, 279-303. Poppel, F. van & I. Joung (2001). Long-term trends in marital status mortality dif-
ferences in The Netherlands 1850-1970. Journal of Biosocial Science, 33, 279-303. 235 Poppel, F. van, J. Schellekens & A.C. Liefbroer (2002). Religious differentials in
infant and child mortality in Holland. Population Studies, 56, 277-290. Power, C., K. Atherton, D.P. Strachan, P. Shepherd, E. Fuller, A. Davis, I. Gibb, M. Kumari, G. Lowe, G.J. Macfarlane, J. Rahi, B. Rodgers & S. Stansfeld (2007). Life-course influences on health in British adults: effects of socio-economic
position in childhood and adulthood. International Journal of Epidemiology, 36,
532-539. Preston, S.H.& M.R. Haines (1991). Fatal years. Child mortality in late nineteenth-centu-
ry America. Princeton: Princeton University Press. Programmacommissie Sociaal-Economische Gezondheidsverschillen. (1989). Een
onderzoeksprogramma voor de periode 1989 t/m 1993. Rijswijk: Ministerie van
WVC. Razzell, P. & C. Spence (2006). The hazards of wealth: adult mortality in pre-twen-
tieth-century England. Social History of Medicine, 19, 381-405. Reek, J. van (1985). Sterfte naar sociale klasse bij volwassenen in Nederland sinds
de negentiende eeuw. Bevolking en gezin, 2, 179-190. Reek, J. van (1993). Mortality by social class among males in the Netherlands
since the nineteenth century. Genus, 49 (1/2), 159-164. Riley, J.C. (1997). Sick, not dead: the health of British workingmen during the mortality
decline. Baltimore: Johns Hopkins University Press. Riley, J.C. (1999). Why sickness and death rates do not move parallel to one
another over time. Social History of Medicine, 12, 101-124. Robert, S. (1999). Socioeconomic position and health: The independent contri-
bution of community socioeconomic context. Annual Review of Sociology, 25,
489-516. Rogers Hollingsworth, J. (1981). Inequality in levels of health in England and
Wales, 1891-1971. Journal of Health and Social Behavior, 22, 268-283. Romijn, W. (1955). Welvaart en gezondheid. Amsterdam: Ziekenzorg. Sandberg, L.G. & R.H. Steckel (1997). Was industrialization hazardous to your
health? Not in Sweden! Literatuur In R.H. Steckel & R. Floud (red.), Health and welfare
during industrialization (pp. 127-160). Chicago: The University of Chicago Press. Siannis, F., J. Copas & G. Lu (2005). Sensitivity analysis for informative censoring
in parametric survival models. Biostatistics, 6, 77-91. Sitter, W. de (1856). Hygiëne en Economie. Tijdschrift voor het Armwezen, 4, 21-34 Smith, D.S. (1983). Differential mortality in the United States before 1900. Journal
of Interdisciplinary History, 13, 735-759. Spree, R. (1988). Health and social class in Imperial Germany. A social history of morta-
lity, morbidity and inequality. Oxford: Berg Publishers. Tijn, T. van (1977). Het sociale leven in Nederland 1844-1873, Algemene Geschiedenis
der Nederlanden (Vol. 12, pp. 131-166). Haarlem: Fibula. Tijn, T. van (1977). Het sociale leven in Nederland 1844-1873, Algemene Geschiedenis
der Nederlanden (Vol. 12, pp. 131-166). Haarlem: Fibula. sociale klasse, sociale mobiliteit en sterfte in nederland Townsend, P. & N. Davidson (1988). Inequalities in health: The Black report and the
Health Divide. London: Penguin Books. Tsuya, N.O. & P. Nystedt (2004). Old-age mortality. In T. Bengtsson, C. Campbell &
J. Z. Lee (red.), Life under pressure. Mortality and living standards in Europe and Asia,
1700-1900 (pp. 399-429). Cambridge, Ma.: The MIT Press. 236 Valkonen, T. (1993). Trends in regional and socioeconomic mortality differentials
in Finland. International Journal of Health Sciences, 3, 157-166. Van de Putte, B. & A. Miles (2005). A social classification scheme for histori-
cal occupational data: partner selection and industrialism in Belgium and
England, 1800-1918. Historical Methods, 38, 61-92. Wadsworth, M.E.J. (1986). Serious illness in childhood and its association with
later-life achievement. In R.G. Wilkinson (red.), Class and health: research and
longitudinal data (pp. 1-20). London: Tavistock. Weir, D.R. (1997). Economic welfare and physical well-being in France, 1750-1990. In R.H. Steckel & R. Floud (red.), Health and welfare during industrialization (pp. 161-200). Chicago: The University of Chicago Press. Wetenschappelijke Raad voor het Regeringsbeleid. (1987). De ongelijke verdeling van
gezondheid (Vol. No. 58). ’s-Gravenhage: Staatsuitgeverij. Whitehead, M. (1998). Diffusion of ideas on social inequalities in health: A Euro-
pean perspective. The Milbank Quarterly, 76, 469-492. Winship, C. & R.D. Mare (1992). Models for sample selection bias. Annual Review of
Sociology, 18, 327-350. Wintle, M. (2000). An economic and social history of the Netherlands, 1800-1920. Demo-
graphic, economic and social transition. Cambridge: Cambridge University Press. Woods, B. (2004). The origins of social class mortality differentials. In P. Boyle, S. Curtis, T. Gatrell, E.G. Literatuur Graham & E. Moore (red.), The geography of health inequa-
lities in the developed world (pp. 37-52). London: Ashgate. Woods, R. & N. Williams (1995). Must the gap widen before it can be narrowed? Long-term trends in social class mortality differentials. Continuity and Change,
10, 105-137. Yen, I.H. & S.L. Syme (1999). The social environment and health: a discussion of
the epidemiologic literature. Annual Review of Public Health, 20, 287-308. Zanden, J.L. van & A. van Riel (2004). The strictures of inheritance: the Dutch economy
in the nineteenth century. Princeton: Princeton University Press. frans van poppel en ruben van gaalen frans van poppel en ruben van gaalen Over de auteurs Onno Boonstra is verbonden aan de afdeling Geschiedenis van de Radboud
Universiteit Nijmegen. Zijn onderzoek richt zich op de sociale geschiedenis van
Nederland in de negentiende eeuw en op de toepassing van statistiek en infor-
matietechnologie in historisch onderzoek, meer in het bijzonder op het gebruik
van geografische informatiesystemen. o.boonstra@let.kun.nl Hilde Bras is als universitair docent verbonden aan de afdeling Methoden en
Technieken van de faculteit der Sociale Wetenschappen van de Vrije Universiteit
Amsterdam. Van 2004 tot en met 2008 doet zij met een VENI-subsidie (NWO Ver-
nieuwingsimpuls) onderzoek naar de invloed van broers en zussen op de levens-
loop in de negentiende eeuw en de eerste helft van de twintigste eeuw. Daar-
naast heeft zij onderzoek gedaan naar de rol van het beroep van dienstbode in de
jongvolwassenheid van vrouwen in het verleden. h j b
@f
l Cees Elzinga is hoofd van de afdeling Methoden en Technieken van de faculteit
der Sociale Wetenschappen van de Vrije Universiteit en participeert in het onder-
zoeksprogramma Comparative Stratification Research. Hij is gespecialiseerd in
modellen voor categorische tijdreeksen. Recent publiceerde hij over metrische
representaties van categorische tijdreeksen en over latente Markov-ketens. ch.elzinga@fsw.vu.nl Theo Engelen is hoogleraar Historische Demografie aan de Radboud Universiteit
Nijmegen en onderzoeksleider bij het N.W. Posthumus Instituut. Zijn onderzoek
richt zich op de demografische geschiedenis van Nederland en Europa in de
negentiende en twintigste eeuw, alsmede de vergelijking daarvan met de ont-
wikkelingen in Taiwan/China. th.engelen@let.ru.nl Ruben van Gaalen studeerde Sociologie aan de Universiteit Bremen. Van sep-
tember 2002-2006 werkte hij bij het Nederlands Interdisciplinair Demografisch
Instituut (NIDI) te Den Haag. In oktober 2007 verdedigde hij zijn proefschrift
getiteld Solidarity and ambivalence in parent-child relationships aan de Universiteit
Utrecht (ICS). Sinds oktober 2006 is hij werkzaam bij het Centraal Bureau voor
de Statistiek (CBS) en verricht onderzoek met behulp van het Sociaal Statistisch
Bestand (SSB). igan@cbs.nl 238 Matthijs Kalmijn is hoogleraar Sociologie aan de Universiteit van Tilburg. Zijn
onderzoek gaat over (a) huwelijksrelaties en echtscheiding, (b) intergeneratio-
nele relaties en overdracht, en (c) etnische verhoudingen en ongelijkheid. Hij
is betrokken bij verschillende grootschalige dataverzamelingsprojecten zoals de
Netherlands Kinship Panel Study en de Panel Study of Social and Cultural Dynamics. Met
Paul de Graaf publiceerde hij over echtscheiding in Nederland in Amerikaanse
tijdschriften zoals Journal of Marriage and the Family, Social Forces en Journal of Fami-
ly Issues. Over de auteurs m.kalmijn@uvt.nl Hans Knippenberg is hoogleraar Politieke en Culturele Geografie aan de Univer-
siteit van Amsterdam. Zijn publicaties liggen vooral op het snijvlak van geschie-
denis en geografie, zoals De eenwording van Nederland (1988; met De Pater), De religi-
euze kaart van Nederland (1992) en The changing religious landscape of Europe (2005). h.knippenberg@uva.nl Jan Kok is senior onderzoeker bij de Virtual Knowledge Studio (Koninklijke
Nederlandse Akademie van Wetenschappen) te Amsterdam en gastdocent bij het
Centrum voor Sociologisch Onderzoek van de Katholieke Universiteit Leuven. Hij houdt zich onder andere bezig met online (globale) samenwerkingsverban-
den tussen sociale en economische historici. Daarnaast verricht hij onderzoek in
de historische demografie en gezinsgeschiedenis. jan.kok@vks.knaw.nl Jan Kok is senior onderzoeker bij de Virtual Knowledge Studio (Koninklijke
Nederlandse Akademie van Wetenschappen) te Amsterdam en gastdocent bij het
Centrum voor Sociologisch Onderzoek van de Katholieke Universiteit Leuven. Hij houdt zich onder andere bezig met online (globale) samenwerkingsverban-
den tussen sociale en economische historici. Daarnaast verricht hij onderzoek in
de historische demografie en gezinsgeschiedenis. jan.kok@vks.knaw.nl Marco H.D. van Leeuwen onderzoekt sociale ongelijkheid en mobiliteit vanaf
1500. Hij werkt als senior onderzoeker op het Internationaal Instituut voor Soci-
ale Geschiedenis en als bijzonder hoogleraar Historische Sociologie bij de Uni-
versiteit Utrecht. Hij onderzoekt ook de geschiedenis van de filantropie, zie V. Kingma en M.H.D. van Leeuwen, Filantropie in Nederland. Voorbeelden uit de peri-
ode 1770-2020 (Aksant 2007), en Giving in the Golden Age (GIGA), http://www.iisg.nl/
research/giga.php. mle@iisg.nl over de auteurs Aart Liefbroer is hoofd van de afdeling Sociale Demografie van het Nederlands
Interdisciplinair Demografisch Instituut in Den Haag en bijzonder hoogleraar
Demografie van Jong-Volwassenen en Intergenerationele Overdracht aan de Vrije
Universiteit Amsterdam. Hij bestudeert vooral ontwikkelingen in de transitie
naar volwassenheid, zowel in Nederland als in Europa. liefbroer@nidi nl 239 Ineke Maas is universitair hoofddocent bij de afdeling Sociologie/ICS van de Uni-
versiteit Utrecht. Zij doet onderzoek naar verschillen tussen landen en histori-
sche veranderingen in sociale ongelijkheid en mobiliteit en publiceerde tevens
over onderwijsongelijkheid en de economische integratie van migranten. Ineke Maas is universitair hoofddocent bij de afdeling Sociologie/ICS van de Uni-
versiteit Utrecht. Zij doet onderzoek naar verschillen tussen landen en histori-
sche veranderingen in sociale ongelijkheid en mobiliteit en publiceerde tevens
over onderwijsongelijkheid en de economische integratie van migranten. over de auteurs Over de auteurs i.maas@uu.nl i.maas@uu.nl Kees Mandemakers is als senior onderzoeker verbonden aan het Internationaal
Instituut voor Sociale Geschiedenis (IISG) en als bijzonder hoogleraar Grote his-
torische databestanden aan de Erasmus Universiteit Rotterdam. Op het IISG
geeft hij leiding aan de Historische Steekproef Nederlandse bevolking (HSN). Hij
publiceerde onder andere over de sociale geschiedenis van het voortgezet onder-
wijs, sociale stratificatie en mobiliteit, zuigelingensterfte en methodologische
aspecten betreffende de verzameling en verwerking van historische onderzoeks-
gegevens. kma@iisg.nl kma@iisg.nl Frans van Poppel is als onderzoeker verbonden aan het Nederlands Interdisci-
plinair Demografisch Instituut (NIDI/KNAW) in Den Haag, een instituut van de
KNAW. Hij promoveerde in 1992 (cum laude) aan de Landbouwuniversiteit Wage-
ningen (Trouwen in Nederland. Een historisch-demografische studie van de 19e en vroeg-
20e eeuw). Zijn onderzoek richt zich op de ontwikkeling van de volksgezondheid,
familie, huwelijk, verwantschap en voortplanting in de negentiende en vroege
twintigste eeuw. Een recente publicatie is Renzo Derosas & Frans van Poppel
(red.), Religion and the Decline of Fertility in the Western World. New York etc.: Sprin-
ger 2006. poppel@nidi.nl Jan Van Bavel doceert demografie en methoden en technieken van maatschap-
pijwetenschappelijk onderzoek aan de Vrije Universiteit Brussel. Hij is hoofd van
de onderzoeksgroep Interface Demography van de Brusselse universiteit en doet er
onderzoek naar de evolutie op lange termijn van vruchtbaarheid en reproductie
in Europa. jvbavel@vub.ac.be over de auteurs Sjoerd de Vos studeerde wiskunde aan de Universiteit van Amsterdam, met als
hoofdrichting statistiek. Vanaf 1970 is hij werkzaam bij (de voorgangers van) de
afdeling Geografie, Planologie en Internationale Ontwikkelingsstudies van deze
universiteit. Bij zijn onderzoek richt hij zich vooral op het terrein van segregatie
en omgevingseffecten. d
@
l 240 s.devos@uva.nl s.devos@uva.nl Richard Zijdeman is promovendus aan de Universiteit Utrecht bij de capaciteits-
groep Sociologie. Hij onderzoekt de effecten van onder meer industrialisering,
massacommunicatie en massatransport op het statusverwervingsproces in Zee-
land in de negentiende en begin twintigste eeuw. In het kader van dit project
heeft hij samen met andere onderzoekers een historische schaal voor beroepssta-
tus ontwikkeld: HIS-CAM. Naast deze onderwerpen, gaat zijn interesse uit naar
collectieve goederen bij het gebruik van moderne communicatiemiddelen, zoals
veilingsites en collaboratories. r.l.zijdeman@uu.nl over de auteurs De afgelopen vijftig jaar zijn de levenslopen van Nederlandse
mannen en vrouwen steeds diverser geworden. Dat betekent
echter geenszins dat de periode daarvoor werd gekenmerkt
door eenvormigheid en onveranderlijkheid. Over de auteurs Toonaangevende
onderzoekers beschrijven in Honderdvijftig jaar levenslopen –
De Historische Steekproef Nederlandse bevolking de levenslopen
van mannen en vrouwen geboren vanaf 1850 en onderzoeken
daarnaast de oorzaken van diversiteit. Aspecten die aan de
orde komen zijn trajecten naar volwassenheid, kinderloosheid,
echtscheiding, religie, analfabetisme, sociale mobiliteit en sterfte. Voor dit onderzoek is gebruik gemaakt van de Historische
Steekproef Nederlandse bevolking. Ineke Maas en Marco van Leeuwen zijn verbonden aan de afdeling
Sociologie van de Universiteit Utrecht en Kees Mandemakers
aan de Faculteit der Historische en Kunstwetenschappen van
de Erasmus Universiteit Rotterdam. Marco van Leeuwen en
Kees Mandemakers zijn tevens werkzaam bij het Internationaal
Instituut voor Sociale Geschiedenis te Amsterdam. Met bijdragen van Jan Van Bavel, Jan Kok & Theo Engelen;
Matthijs Kalmijn; Hans Knippenberg & Sjoerd de Vos; Onno
Boonstra; Richard L. Zijdeman & Kees Mandemakers; Ineke Maas
& Marco van Leeuwen; Frans van Poppel & Ruben van Gaalen;
Hilde Bras, Aart C. Liefbroer & Cees H. Elzinga. ISBN 978 90 8964 067 3
|
https://openalex.org/W2793399621
|
https://hal.archives-ouvertes.fr/hal-01764937/file/fenrg-06-00011.pdf
|
English
| null |
Nanometrology of Biomass for Bioenergy: The Role of Atomic Force Microscopy and Spectroscopy in Plant Cell Characterization
|
Frontiers in energy research
| 2,018
|
cc-by
| 8,827
|
Review Review
published: 19 March 2018
doi: 10.3389/fenrg.2018.00011 published: 19 March 2018
doi: 10.3389/fenrg.2018.00011 Nanometrology of Biomass for
Bioenergy: The Role of Atomic Force
Microscopy and Spectroscopy in
Plant Cell Characterization Anne M. Charrier1, Aude L. Lereu2, Rubye H. Farahi3, Brian H. Davison4 and
Ali Passian3,4,5,6* Anne M. Charrier1, Aude L. Lereu2, Rubye H. Farahi3, Brian H. Davison4 and
Ali Passian3,4,5,6* 1 Aix Marseille Univ, CNRS, CINaM, Marseille, France, 2 Aix Marseille Univ, CNRS, Centrale Marseille, Institut Fresnel,
Marseille, France, 3 Quantum Information Science, Computational Sciences and Engineering Division, Oak Ridge National
Laboratory, Oak Ridge, TN, United States, 4 BioEnergy Science Center (BESC), Biosciences Division, Oak Ridge National
Laboratory, Oak Ridge, TN, United States, 5 Department of Chemical and Biomolecular Engineering, University of Tennessee,
Knoxville, TN, United States, 6 Department of Physics, University of Tennessee, Knoxville, TN, United States Ethanol production using extracted cellulose from plant cell walls (PCW) is a very promis-
ing approach to biofuel production. However, efficient throughput has been hindered by
the phenomenon of recalcitrance, leading to high costs for the lignocellulosic conversion. To overcome recalcitrance, it is necessary to understand the chemical and structural
properties of the plant biological materials, which have evolved to generate the strong
and cohesive features observed in plants. Therefore, tools and methods that allow the
investigation of how the different molecular components of PCW are organized and
distributed and how this impacts the mechanical properties of the plants are needed but
challenging due to the molecular and morphological complexity of PCW. Atomic force
microscopy (AFM), capitalizing on the interfacial nanomechanical forces, encompasses
a suite of measurement modalities for nondestructive material characterization. Here,
we present a review focused on the utilization of AFM for imaging and determination
of physical properties of plant-based specimens. The presented review encompasses
the AFM derived techniques for topography imaging (AM-AFM), mechanical properties
(QFM), and surface/subsurface (MSAFM, HPFM) chemical composition imaging. In
particular, the motivation and utility of force microscopy of plant cell walls from the early
fundamental investigations to achieve a better understanding of the cell wall architecture,
to the recent studies for the sake of advancing the biofuel research are discussed. An
example of delignification protocol is described and the changes in morphology, chemi-
cal composition and mechanical properties and their correlation at the nanometer scale
along the process are illustrated. Edited by:
Abdul-Sattar Nizami,
King Abdulaziz University,
Saudi Arabia
Reviewed by:
Héctor A. Ruiz,
Universidad Autónoma de
Coahuila, Mexico
Mohammad Rehan,
King Abdulaziz University,
Saudi Arabia
*Correspondence:
Ali Passian
passianan@ornl.gov Edited by:
Abdul-Sattar Nizami,
King Abdulaziz University,
Saudi Arabia Reviewed by:
Héctor A. Ruiz,
Universidad Autónoma de
Coahuila, Mexico
Mohammad Rehan,
King Abdulaziz University,
Saudi Arabia *Correspondence:
Ali Passian
passianan@ornl.gov Specialty section:
This article was submitted to
Bioenergy and Biofuels,
a section of the journal
Frontiers in Energy Research Received: 26 October 2017
Accepted: 22 February 2018
Published: 19 March 2018 Keywords: AFM, QFM, MSAFM, HPFM, plant cell walls, lignin, biofuel, bioenergy Keywords: AFM, QFM, MSAFM, HPFM, plant cell walls, lignin, biofuel, bioenergy 1. INTRODUCTION Such advances in biomechanics have
been possible due to the development of new technologies allow-
ing mechanical and chemical analysis of surfaces with submicro-
nic or nanometric lateral resolution (Gindl and Schoberl, 2004;
Beecher et al., 2009; Tetard et al., 2011; Burgert and Keplinger,
2013). Such methodologies used to map the various properties of
plants at the submicron scale are reported in Table 1. of recalcitrance, cellulose extraction is challenging. Overcoming
the chemical and structural properties that have evolved in
biomass to form strong and cohesive structures hence requires
a deep understanding of how the molecular composition and
organization in PCW impacts the mechanical properties of the
plants. Indeed, the correlation between the molecular traits of
the cell walls and its morphological and mechanical charac-
teristics is ultimately expressed in the plant response. Due to
their hierarchical structure, the macroscopic appearance and
performance of plants have been shown to be inevitably linked
to their structure at the micro- and nanometer scales (Gibson,
2012). However, the nanoscale behavior of many physical quan-
tities, such as elasticity and plasticity, intimately connected with
such a correlation, is largely unknown or poorly understood. Fundamentally, this inevitably leads to questioning the relation-
ship between chemistry, structural organization and mechanical
properties and how such molecular scale variations in the physi-
cal quantities of the cell walls translate into the bulk level and
on to the whole organism level. From a practical point of view,
this leads to questioning how cell wall chemical composition
can be altered to intrinsically modify wood properties (Gindl
and Gupta, 2002; Pilate et al., 2002), for example, to better suit
various applications. Such complexity and heterogeneity of the
plant systems therefore question the relevance of macroscopic
and microscopic measurements. Often, these can only give an
average description of the properties of the considered object. In
recent years, many mechanical studies, primarily addressing the
cell and cell wall level of plant body, have enlightened important
structure–property and structure–function relationships (Fratzl
and Weinkamer, 2007). Such advances in biomechanics have
been possible due to the development of new technologies allow-
ing mechanical and chemical analysis of surfaces with submicro-
nic or nanometric lateral resolution (Gindl and Schoberl, 2004;
Beecher et al., 2009; Tetard et al., 2011; Burgert and Keplinger,
2013). Such methodologies used to map the various properties of
plants at the submicron scale are reported in Table 1. 1. INTRODUCTION As an example, the method of nanoindentation has proven to
be very useful in providing important mechanical information
(Gindl and Gupta, 2002; Zickler et al., 2006). This technique
pushes a microscopic indenter with a known shape into a
material surface while continuously measuring the response of
the tip-sample system: the loading and unloading forces, the
penetration depth, and the surface restoration to the indentation. Since forces ranging from tens of nanonewtons to millinewtons
with submicrometer lateral resolutions can be precisely applied,
the nanoindentation testing cycle can be highly sensitive to the
mechanical properties of the material. The extent of the material
restoration following the penetration into the surface is a measure
of the material’s elasticity, which is generally quantified by the
elastic modulus or Young’s modulus (E). Likewise, if any perma-
nent indentation occurs, the material displays plasticity, which is
quantified by the plasticity index (PI). The pull-off force when the
tip is retreated from the surface is related to the adhesion energy,
which are due to attractive van der Waals and electrostatic forces
and other attractive forces acting between the tip and sample. In
one of the first nanoindentation experiments, Wimmer and Lucas
(1997) explored the relationship between hardness and Young’s
moduli for the secondary cell wall and cell corner middle lamella
in spruce wood. They reported that the mechanical properties of
secondary walls along their longitudinal direction is correlated
with the presence of organized unidirectional cellulosic fibrils
(Wimmer et al., 1997) while the reduced elastic modulus in the
lignin-rich middle lamella was associated with the absence of
cellulose in the region (Wimmer and Lucas, 1997). It was also
demonstrated that the elastic modulus in the secondary walls
decreased with microfibril angle therefore showing the impor-
tance of molecular organization (Gindl et al., 2004). These studies
were the first to demonstrate the relationship between chemical
composition, molecular organization and mechanical properties. However, in addition to requiring sample processing (polishing)
and embedding into a polymeric material to avoid distortion of
the measurements due to surface roughness (Konnerth et al.,
2008; Burgert and Keplinger, 2013; Wagner et al., 2014;
Youssefian et al., 2017), nanoindentation is ultimately limited by
the resolution of the apparatus. Furthermore, the measurements
are discrete, and do not allow mapping of mechanical properties. TABLE 1 | Available methodologies for mapping the structure, the chemical
composition, and the mechanical properties of plants at the submicron scale. 1. INTRODUCTION Charrier AM, Lereu AL, Farahi RH,
Davison BH and Passian A (2018)
Nanometrology of Biomass for
Bioenergy: The Role of Atomic Force
Microscopy and Spectroscopy in
Plant Cell Characterization. Front. Energy Res. 6:11. doi: 10.3389/fenrg.2018.00011 An important scientific focus and challenge of the 21st century is to achieve non-food biofuel
mass production and thus replace fossil fuels (Lynd et al., 2008; Sannigrahi et al., 2010; Mohapatra
et al., 2017). Significant research is being dedicated to ethanol production using extracted cel-
lulose from the plant cell walls (PCW). Due to the complex structure of the PCW, which contain a
variety of organic components (cellulose, hemicellulose, lignin, pectin, etc.), and the phenomenon March 2018 | Volume 6 | Article 11 Frontiers in Energy Research | www.frontiersin.org 1 AFM on Plant Cell Walls? Charrier et al. of recalcitrance, cellulose extraction is challenging. Overcoming
the chemical and structural properties that have evolved in
biomass to form strong and cohesive structures hence requires
a deep understanding of how the molecular composition and
organization in PCW impacts the mechanical properties of the
plants. Indeed, the correlation between the molecular traits of
the cell walls and its morphological and mechanical charac-
teristics is ultimately expressed in the plant response. Due to
their hierarchical structure, the macroscopic appearance and
performance of plants have been shown to be inevitably linked
to their structure at the micro- and nanometer scales (Gibson,
2012). However, the nanoscale behavior of many physical quan-
tities, such as elasticity and plasticity, intimately connected with
such a correlation, is largely unknown or poorly understood. Fundamentally, this inevitably leads to questioning the relation-
ship between chemistry, structural organization and mechanical
properties and how such molecular scale variations in the physi-
cal quantities of the cell walls translate into the bulk level and
on to the whole organism level. From a practical point of view,
this leads to questioning how cell wall chemical composition
can be altered to intrinsically modify wood properties (Gindl
and Gupta, 2002; Pilate et al., 2002), for example, to better suit
various applications. Such complexity and heterogeneity of the
plant systems therefore question the relevance of macroscopic
and microscopic measurements. Often, these can only give an
average description of the properties of the considered object. In
recent years, many mechanical studies, primarily addressing the
cell and cell wall level of plant body, have enlightened important
structure–property and structure–function relationships (Fratzl
and Weinkamer, 2007). 1. INTRODUCTION Plant
property
Mapping technique
Lateral
resolution
Reference
Structure
Scanning electron
microscopy
10 nm
Auxenfans et al. (2017)
Atomic force microscopy
1 nm
Salvadori et al. (2014)
Chemical
composition
ToF-SIMS
1 µm
Tolbert and Ragauskas
(2017)
Confocal Raman
microscopy
300 nm
Tetard et al. (2015)
Mode synthesized AFM
1 nm
Tetard et al. (2011)
Hybrid photonic force
microscopy
5 nm
Farahi et al. (2017)
Mechanical
properties
Nanoindentationa
<1 μm
Wimmer and Lucas
(1997)
Quantitative force–volume
mapping
10 nm
Farahi et al. (2017)
aNanoindentation usually provides discrete measurements. 2
Frontiers in Energy Research | www.frontiersin.org TABLE 1 | Available methodologies for mapping the structure, the chemical
composition, and the mechanical properties of plants at the submicron scale. More recently, atomic force microscopy (AFM) and its deriva-
tive techniques have emerged as the method of choice to resolve
cell wood properties at the nanometer scale. The first force meas-
urements with AFM also invoked a nanoindenter, but with an
improved lateral resolution down to a few nanometers (Kirby, 2011;
Fernandes et al., 2012). More recently, quantitative force–volume
mapping (QFM) was adapted to plant cell wall measurements uti-
lizing continuous force curve recording and therefore solving the
problem of heterogeneity observation by imaging the topography
with the associated E and PI mappings. Studies on Arabidopsis
thaliana reported different characteristic modes of deformation
and a spatial distribution of the elastic moduli across the surface
(Yakubov et al., 2016), while Radotic et al. (2012) showed the
changes in stiffness of the cell walls at different phases of growth. In addition, new innovative AFM techniques such as mode syn-
thesized AFM (MSAFM) (Tetard et al., 2010, 2011) and hybrid
photonic force microscopy (HPFM) (Tetard et al., 2015) have been March 2018 | Volume 6 | Article 11 2 2 Charrier et al. AFM on Plant Cell Walls? The chemical processing of the biomass, here in the form of
cross-sections (left image of Figure 1), was intended to remove
lignin and other compounds to facilitate a more efficient extraction
of the cellulose. The samples were received, frozen, and sectioned
into 20 µm thick sections with a cryotome using a disposable
blade that allowed minimal structural damage. For the untreated
samples, the cross-section were washed with deionized (DI)
water and dried between glass slides, referred as the untreated
raw biomass or fresh Populus (UR). 2. DELIGNIFICATION PROCESSES The extraction of cellulose, while abundant in the secondary
plant cell walls, is particularly hindered by the presence of lignin. Lignin is a complex organic polymer widely recognized for play-
ing a role in the structural integrity of plants. As a compound, it
is mostly inert and nonreactive, and thus makes a good material
candidate for manufacturing and industrial applications (e.g.,
see lignin-based carbon fibers and nanofibers; Fang et al., 2017). Lignin removal has been a central key issue where a whole variety
of complex processes, also referred to as delignification processes,
have been developed (Singh et al., 2014). Controlling this pro-
cess requires the understanding of the relationship between the
chemical composition, the structural aspect, the biological infra-
structure, and the mechanical properties of the PCW. The many
delignification strategies have been based on various approaches
(Singh et al., 2014) such as (1) physical treatments including
mechanical (Cadoche and Lopez, 1989), pyrolysis (Chen et al.,
2016), and steam processes (Tian et al., 2017), (2) chemical using
organosolv (Erdocia et al., 2014), acid and alkaline hydrolysis
(Singh et al., 2015; Carlos Martinez-Patino et al., 2017), or
saccharification and fermentation (Healey et al., 2015), and (3)
biological (Asina et al., 2016) and enzymatic (Al-Zuhair et al.,
2015; Tian et al., 2017) treatments. To illustrate the complexity
of delignification, an example based on a multistep chemical
protocol, is given in Figure 1, resulting in a set of samples to be
investigated using AFM techniques (Farahi et al., 2017). 1. INTRODUCTION Next the extractive-free
(EF) Populus were prepared by removing extractives (inorgan-
ics: K, Ca, Mg, P, F, Na, Si, S, Mn, etc. and organics: terpenes,
fats, waxes, flavanoids, tannins, stilbenes, etc.) by refluxing with
dichloromethane and then dried between glass slides to preserve
the morphology of the sample and avoid contaminations. The
removal of extractives allowed for the cellulose and lignin to be
detected with less contamination on the surface when using ToF-
SIMS, Raman, AFM, and other characterization methods (Farahi
et al., 2017). The EF sample is then subject to glacial acetic acid
combined with sodium chlorite resulting in the extractive free
holopulped (EH) sample with most of the lignin removed. Finally,
hydrochloric acid treatment is imposed on the EH sample to keep
only the cellulose by removing hemicellulose and residuals of
lignin (EHA sample). See details of the protocols in Farahi et al. (2017). The presented delignification process results in sets of
four samples to be investigated. developed. The MSAFM was utilized to obtain the subsurface
nanomorphological properties of wood cell walls with structural
changes along the delignification process of poplar (Tetard et al.,
2010, 2011), while the HPFM correlated the structural changes
with chemical composition (Tetard et al., 2015).h This manuscript aims at demonstrating the input of nanome-
trology using AFM in understanding how the molecular distri-
bution and organization impacts the mechanical strength and
cohesion of PCW to better develop cellulose extraction protocols
through delignification. Therein, the different applications of
AFM modes for plant cell characterization are reviewed within
the context of biofuel research. In Section 2, an example of chemi-
cal protocol leading to delignification starting from a cryotomed
young poplar is described. In Section 3, the operating principle
of AM-AFM, QFM, MSAFM/HPFM modes are described and
their use to extract structural, mechanical, and chemical informa-
tion through the process of delignification is illustrated through
examples. 3.1. Amplitude Modulation AFM (AM-AFM):
Structural Properties p
AFM is based on measuring the interaction between a sharp tip
(radius of curvature of a few nanometers), mounted on a canti-
lever spring (0.01 N/m < k < 150 N/m), and a surface; when the
tip is raster-scanned across the surface, a high spatial resolution
topographical image may be obtained (Binnig et al., 1986). As the
tip comes near the surface, interaction forces induce a bending
of the cantilever, which for small amplitudes follows Hooke’s law. Depending on the environment, the tip/sample distance and the
mode of utilization, the forces associated with AFM include van
der Waals, electrostatic, magnetic, capillary, contact mechanical
forces, chemical bonding, etc. Forces as small as few tens of picone-
wton have been measured. AFM offers the possibility of imaging
the surface of any material (hard, soft, insulators, heterogeneous)
in a variety of environments (solution, air, vacuum, or gas). Among p
AFM is based on measuring the interaction between a sharp tip
(radius of curvature of a few nanometers), mounted on a canti-
lever spring (0.01 N/m < k < 150 N/m), and a surface; when the
tip is raster-scanned across the surface, a high spatial resolution
topographical image may be obtained (Binnig et al., 1986). As the
tip comes near the surface, interaction forces induce a bending
of the cantilever, which for small amplitudes follows Hooke’s law. FIGURE 1 | Chemical steps for delignification process starting from a cryotomed young poplar. AFM studies are carried out at each step of the process, i.e., on
untreated raw (UR), extractive-free (EF), holopulped (EH), and acid-treated (EHA) samples (adapted from Farahi et al. (2017)). FIGURE 1 | Chemical steps for delignification process starting from a cryotomed young poplar. AFM studies are carried out at each step of the process, i.e., on
untreated raw (UR), extractive-free (EF), holopulped (EH), and acid-treated (EHA) samples (adapted from Farahi et al. (2017)). March 2018 | Volume 6 | Article 11 Frontiers in Energy Research | www.frontiersin.org 3 AFM on Plant Cell Walls? Charrier et al. the basic modalities of AFM, amplitude-modulation atomic force
microscopy (AM-AFM) (Martin et al., 1987; Zhong et al., 1993;
Garcia and Perez, 2002; Garcia et al., 2007), also called tapping
mode, has provided large improvements to soft materials imaging
in general due to its non-invasive and non-destructive operation,
and molecular resolution. 3.1. Amplitude Modulation AFM (AM-AFM):
Structural Properties The cantilever is approximated as a
linear elastic spring and its bending, D(z), is related to the applied
loading force of the indentation, P, and k the spring constant of
the cantilever according to Hooke’s law, P = −kD(z). AM-AFM has been successful in characterizing biomass
(Kirby et al., 1996; Salvadori et al., 2014; Farahi et al., 2017) and
in particular the architecture of plant cell walls at the molecular
level which in turn is essential in improving lignocellulosic feed-
stock properties (Zhang et al., 2013b, 2017; Keplinger et al., 2014;
Torode et al., 2018). AM-AFM mapping of cellulose topography
revealed the right-handed twisted nature of microfibrils and the
periodic distribution of glucose and fiber unit along the microfi-
brils (Hanley et al., 1997). Near-atomic resolution of the cellulose
structure was reached, revealing the triclinic structure of Iα crystal
phase (Baker et al., 2000). AM-AFM has also been useful in meas-
uring surface morphology and roughness changes after treatment
(Medeiros et al., 2007; Zhang et al., 2013a; Nanda et al., 2015). Nanomechanical information extracted from analysis of
force curve measurements include elastic and dissipative
components. Young’s modulus, E, can be calculated from the
slope of the unloading curve (Pharr et al., 1992) (Figure 3E)
using a suitable model, which depends on the tip shape and
dimensions (Bulichev and Alekhin, 1987), sample dimensions
(flat, spherical) and whether or not dissipative components
such as plastic deformation, adhesion, or viscosity are present
(Johnson et al., 1971; Derjaguin et al., 1975; Butt et al., 2005). Dissipative components such as plastic deformation and adhe-
sion hysteresis are included in the force measured in quasi-
static equilibrium and can be extracted from a force–distance
curve. Determination of non-conservative or velocity-dependent
components requires dynamic force measurements (Herruzo
et al., 2014). g
In Figures 2A–D, the structural changes (morphology and
wall thickness) undergone by the PCW were revealed at each
chemical step of the delignification process detailed above. The
sharper and well-contrasted PCW appearance after exposure to
dichloromethane (EF image of Figure 2B) could be explained
by the removal of organics, leaving the sample surface cleaned
(Farahi et al., 2017). The UR and EF samples appeared very
similar with well-defined PCW preserving the cell structure and
having an average thickness of 3 µm. However, the structures of
EH and EHA samples underwent considerable changes with an
average PCW thickness around 2 µm. 3.1. Amplitude Modulation AFM (AM-AFM):
Structural Properties Lateral resolution is mostly determined
by the tip radius of curvature while subnanometric vertical
resolution is provided by the piezoelectric modulus and feedback
regulation used to control vertical displacements. In AM-AFM, the
cantilever is mechanically excited near or at its intrinsic resonant
frequency (typically in the range from a few tens to a few hundreds
of kHz) with a given amplitude. When the tip is brought to within
a few nanometers above the surface, the tip-sample interaction
changes the resonance frequency of the cantilever and leads to a
decrease of its oscillation amplitude whether the forces involved
are attractive or repulsive. To measure the force intensity and its
nature (attractive or repulsive), a laser beam (in most systems)
is generally positioned at the apex of the cantilever and reflected
to a position-sensing photodiode. The bending of the cantilever
induces a deflection of the laser resulting in a displacement of the
reflected beam on the photodiode. The new position is then used
to apply a retroactive feedback to maintain the amplitude of oscil-
lations (referred as a setpoint) of the cantilever. mapping (QFM) (Radmacher et al., 1996) and peak-force
(Adamcik et al., 2011) allow nanomechanical mapping with
nanometer scale lateral resolution, and measurements of forces
down to few piconewtons. These techniques are currently used
for mechanical characterization in cell biology (living cells) and
structural biology, including testing of cartilage (Heu et al., 2012),
bones (Spitzner et al., 2015), soft tissues (Burgert and Keplinger,
2013), and wood (Farahi et al., 2017). Similar to nanoindentation,
the main principle of force measurements is to calculate hardness
and elastic modulus from a load–displacement curve recorded
during a local indentation (Figures 3B,D–F). In a single force curve, a local force measurement is realized
at a given position, corresponding to one pixel of the image, by
only recording the corresponding loading and unloading curves. In quantitative force–volume mapping (Radmacher, 1997), single
force curves are measured at points on a 2D grid. Tip-sample
force is controlled by discrete force triggering at each point. Peak-
force is an advanced version of QFM in which the force curves
are acquired simultaneously with the topography mapping
(Adamcik et al., 2011; Durkovic et al., 2014).l During an indentation measurement, laser deflection versus
vertical (z) displacement is recorded continuously. In order to
extract useful quantities, the data are converted into force versus
tip/sample distance curves. 3.1. Amplitude Modulation AFM (AM-AFM):
Structural Properties With the lignin content
removed in EHA, the PCW started to collapse and the overall
cell sizes became smaller and compressed. The decrease in the
average thickness of PCW (Figure 2E, gray) along the process
was in agreement with the lignin removal. In addition to the Young’s modulus, a plasticity index (PI),
including plasticity and viscosity components, can be extracted. From each indentation curve, the area comprised between the
loading and the unloading curves above the zero force line, A1, is a
measure of the energy needed for the deformation and dissipated
into the sample, whereas the sum A1 + A2 is the maximal energy
that could be stored in the sample during the indentation (Butt
et al., 2005), with A2 being the area below the unloading curve (see
Figure 3F). The plasticity index can then be defined as PI = A1/
(A1 + A2) and can be used to discriminate components with dif-
ferent viscoplasticity. If PI = 1, the material is fully viscoplastic. If
PI = 0, it indicates a perfectly elastic behavior. Finally, adhesion
energy can also be extracted for the unloading curve as illustrated
in Figure 3E, where the A0 area shows the presence of adhesion
between the surface and the AFM probe. 3.2. Quantitative Force–Volume Mapping
(QFM): Mechanical Properties Force measurements using AFM (Zdunek and Kurenda, 2013)
and derivative technologies such as quantitative force–volume March 2018 | Volume 6 | Article 11 Frontiers in Energy Research | www.frontiersin.org 4 AFM on Plant Cell Walls? Charrier et al. FIGURE 2 | Examination of poplar samples through the delignification process steps. (A–D) AM-AFM topographic images of untreated raw (UR), extractive-free (EF),
holopulped (EH) and acid-treated (EHA) samples showing the structural changes along the delignification process (image size 50 × 50 µm2). The average PCW
thicknesses extracted from the topographic images are plotted in (E) (gray curve). (e) Images: Young’s modulus (GPa) mappings for each type of samples extracted
from quantitative force–volume mappings. Image size is 5 µm × 5 µm. The corresponding average Young’s moduli values (E) are reported in red. Blue triangles:
Average values of the plasticity index (PI) extracted from the quantitative force–volume mapping. Data from Farahi et al. (2017), revisited. FIGURE 2 | Examination of poplar samples through the delignification process steps. (A–D) AM-AFM topographic images of untreated raw (UR), extractive-free (EF),
holopulped (EH) and acid-treated (EHA) samples showing the structural changes along the delignification process (image size 50 × 50 µm2). The average PCW FIGURE 2 | Examination of poplar samples through the delignification process steps. (A–D) AM-AFM topographic images of untreated raw (UR), extractive-free (EF),
holopulped (EH) and acid-treated (EHA) samples showing the structural changes along the delignification process (image size 50 × 50 µm2). The average PCW
thicknesses extracted from the topographic images are plotted in (E) (gray curve). (e) Images: Young’s modulus (GPa) mappings for each type of samples extracted
from quantitative force–volume mappings. Image size is 5 µm × 5 µm. The corresponding average Young’s moduli values (E) are reported in red. Blue triangles:
Average values of the plasticity index (PI) extracted from the quantitative force–volume mapping. Data from Farahi et al. (2017), revisited. Examples of E (Figure 3B) and PI (Figure 3C) mappings
extracted from an untreated raw poplar sample were correlated
with the topography image in Figure 3A (Farahi et al., 2017). The
E mapping showed strong Young’s modulus heterogeneity. Force
curves shown in Figures 3D–F are discrete curves measured
at the marked points in Figure 3A. They were chosen at three
regions of the sample to illustrate different mechanical behaviors. In Figure 2D, the loading and unloading curves were super-
imposed showing a perfectly elastic behavior. 3.2. Quantitative Force–Volume Mapping
(QFM): Mechanical Properties In Figures 3E,F,
the unloading curve is below the loading curve, translating into
viscoplastic properties. In addition, adhesion between the tip and
the surface during unloading (orange region) was observed in
Figure 3E. The PI values in the mapping of Figure 3C varied
between 0 and 1, therefore indicating regions with strong elastic
or viscoplastic behaviors. As illustrated in Figure 2, the Young’s moduli varied toward
lower values as the lignin was removed with a decrease from
4.7 ± 0.7 GPa for UR to 0.34 ± 0.07 GPa for EHA therefore suggest-
ing total removal of the hemicellulose and lignin from the sample. Interestingly, the average values of PI (blue curve in Figure 2E)
evolving around 0.5 for the UR, EF, and EH samples, revealed an
important difference after acid treatment (EHA) showing a nearly
perfect elastic behavior which did not seem to be directly related
to delignification but mainly to the removal of hemicellulose. Frontiers in Energy Research | www.frontiersin.org 3.3. Mode Synthesizing Atomic Force
Microscopy (MSAFM) and Hybrid Photonic
Force Microscopy (HPFM): Chemical
Compositiont This
intended coupled dynamics assumes that the system possesses
suitable mechanical dispersion and supports a propagation mode
in the megahertz spectrum. In a typical example such as consid-
ered here, two forcings, that is, driving forces with frequencies ω1
and ω2, respectively, are delivered to the probe via piezoelectric
elements driven by waveform generators (Farahi et al., 2017). In both MSAFM and HPFM, the specific type of driving of the
sample and the probe is selected based upon the specific applica-
tion and the need for accessing subsurface, topographical, or
chemical information. In general, both the probe and sample can
be driven (Tetard et al., 2008) via any number and combination
of waveform. In either case, the probe-sample interaction via the
van der Waals force allows for synthesis of new oscillation modes
in the system. For the case considered, two modes in the first
order coupling are generated: a sum frequency of ω+ = ω1 + ω2
and a difference frequency of ω− = |ω1 − ω2|. The response signal
at ω− is of greater interest since it can be set in the kHz range
and be readily monitored with phase-locked loop techniques. In
general, from the effect of various interactions (elastic, dissipa-
tive, etc.), a measurement of the amplitude and phase properties of the probe at the difference frequency, ω− (or other selected
frequencies), high-resolution subsurface images formed by the
contrast due to the variations in elasticity, mechanical losses, etc.,
of the surface and subsurface material domains are constructed. Thus, the contrast measured from the dynamics of the probe can
be used to obtain morphological and chemical distinctions in the
examined materials. To achieve specific compositional informa-
tion, HPFM introduces amplitude modulated infrared light with
a wavelength λ to the sample at the difference frequency, creating
yet another mechanical actuation based on the photothermal
absorption of the sample. Mid-IR quantum cascade lasers (QCL)
are useful excitation sources since they can be modulated and set
at a wavelength to maximize composition-dependent absorption. The induced photothermal effect is exploited, where the absorp-
tion of light decays into heat, which in turn is conducted away as
it dissipates. The resulting thermal expansion and relaxation is
the source of a third mechanical actuation by photonic excitation
at ωQCL → ω–, which is associated with the absorption properties
of the sample. 3.3. Mode Synthesizing Atomic Force
Microscopy (MSAFM) and Hybrid Photonic
Force Microscopy (HPFM): Chemical
Compositiont Quantitative force–volume mapping has been used to study
the nanomechanical properties of plant cell walls at every step of
the delignification process (Peaucelle, 2014; Farahi et al., 2017). Chemical composition is another sought-after parameter when
investigating plant cell walls or biological samples. Simultaneous
mechanical and chemical mapping have been obtained using March 2018 | Volume 6 | Article 11 Frontiers in Energy Research | www.frontiersin.org 5 AFM on Plant Cell Walls? Charrier et al. FIGURE 3 | Topographic (A) and associated mechanical imaging (B,C) (Young’s modulus (E) and plastic index (PI) mappings) (scale bar 1 µm) obtained on an
untreated raw poplar cell wall. (D–F) Discrete force curves extracted at the marked localizations on the topographic image (A). Red and black curves correspond to
the loading and unloading during tip indentation [inset (D)], respectively. (D) gives an illustration of elastic behavior and (E) of plasticity with adhesion (A0). S, the
slope obtained by fitting the beginning of the unloading curve, is used to calculate the Young’s modulus. A1 and A2 in (F) are two parameters used to access the
plasticity index. Data from Farahi et al. (2017), revisited. FIGURE 3 | Topographic (A) and associated mechanical imaging (B,C) (Young’s modulus (E) and plastic index (PI) mappings) (scale bar 1 µm) obtained on an
untreated raw poplar cell wall. (D–F) Discrete force curves extracted at the marked localizations on the topographic image (A). Red and black curves correspond to
the loading and unloading during tip indentation [inset (D)], respectively. (D) gives an illustration of elastic behavior and (E) of plasticity with adhesion (A0). S, the
slope obtained by fitting the beginning of the unloading curve, is used to calculate the Young’s modulus. A1 and A2 in (F) are two parameters used to access the
plasticity index. Data from Farahi et al. (2017), revisited. mode synthesizing atomic force microscopy (MSAFM) (Tetard
et al., 2008, 2010, 2011) and the additional spectroscopic capa-
bilities of hybrid photonic nanomechanical force microscopy
(HPFM) (Tetard et al., 2015; Farahi et al., 2017). Capitalizing on
the nonlinear probe-sample forces, microscopy with MSAFM
offers a way to image soft samples and probe nanostructures that
are below their surfaces (subsurface imaging). Briefly, in MSAFM
multiple mechanical excitations (e.g., with megahertz frequencies)
introduce small mechanical actuations within the sample and
the probe, resulting in a coupled probe-sample dynamics. Frontiers in Energy Research | www.frontiersin.org 3.3. Mode Synthesizing Atomic Force
Microscopy (MSAFM) and Hybrid Photonic
Force Microscopy (HPFM): Chemical
Compositiont HPFM and MSAFM were used in conjunction with confocal
Raman microscopy to characterize the distribution of chemical
species at the nanoscale on poplar cross-sections (Tetard et al.,
2015). Confocal Raman microscopy was first performed over the
cell wall to spatially identify the cellulose-rich and lignin-rich
regions (Figure 4A). Simultaneous AFM (Figure 4B), MSAFM
(Figure 4C), and HPFM (Figure 4D) were then carried out over
the same region. With the MSAFM image, new detail was found
in the lignin-rich region compared to the AFM image, such as the
structures within the dashed circled regions. The MSAFM signal
was also able to detect substructures in the cellulose-rich region. March 2018 | Volume 6 | Article 11 Frontiers in Energy Research | www.frontiersin.org 6 AFM on Plant Cell Walls? Charrier et al. FIGURE 4 | Demonstration of HPFM compared to confocal Raman, AFM, and MSAFM on EF (extractive-free) poplar cell wall. (A) Confocal Raman image showing
lignin-rich (pink) and cellulose-rich (blue, green) regions. (B) Simultaneous AFM image showing only topography. (C) MSAFM image at difference frequency,
ω− = 26 kHz, brings additional clarity to the lignin structures indicated by the dashed circled regions. (D) HPFM image at photonic actuation, ωQCL → ω_ = 26 kHz,
reveals additional detail of the near-surface cellulosic globules shown outlined. Adapted from Tetard et al. (2015). FIGURE 4 | Demonstration of HPFM compared to confocal Raman, AFM, and MSAFM on EF (extractive-free) poplar cell wall. (A) Confocal Raman image showing
lignin-rich (pink) and cellulose-rich (blue, green) regions. (B) Simultaneous AFM image showing only topography. (C) MSAFM image at difference frequency,
ω− = 26 kHz, brings additional clarity to the lignin structures indicated by the dashed circled regions. (D) HPFM image at photonic actuation, ωQCL → ω_ = 26 kHz,
reveals additional detail of the near-surface cellulosic globules shown outlined. Adapted from Tetard et al. (2015). provide a unique opportunity to relate the chemistry (HPFM) to
the structure (AFM and MSAFM) and the mechanical properties
(QFM and Peak-force) of the same regions at the nanometric scale. Understanding how cell wall chemical composition can be altered
to intrinsically modify wood properties is also a highly promising
issue; for example to increase the proportion of cellulose in plant
cell walls, to reduce the lignin content, or to make delignification
an easier process by reducing the elastic modulus of the plant cell
walls. 4. CONCLUSION From the review of the reported case studies, we conclude that the
AFM based measurement science and technology for exploring,
manipulating, understanding, and relating the different proper-
ties of plant cell walls constitutes an emerging area of research
within the plant biological material characterization. Given
the current state of the understanding of the plant cells, there
is a tremendous need for innovative approaches to microscopy
and spectroscopy that can aid, for example, the development of
efficient protocols for polysaccharides extraction in plants and
further bioethanol production. From the considered studies to
date, it is not difficult to form the opinion that understanding
how the different molecular components of the plant cell walls
are intermingled and distributed and how these impacts the
mechanical properties of the plants will help in developing opti-
mized process for overcoming recalcitrance. Indeed, the AFM
techniques, being nondestructive and amenable to operation
under ambient conditions with specimens in their native states, ACKNOWLEDGMENTS We would like to thank David Graham at ORNL for reviewing the
manuscript. This work was sponsored by the BioEnergy Science
Center (BESC) of the Oak Ridge National Laboratory (ORNL). The BESC is a US Department of Energy (DOE) Bioenergy
Research Center supported by the Office of Biological and
Environmental Research in the DOE Office of Science. ORNL is
managed by UT-Battelle, LLC, for the US DOE under contract
DE-AC05-00OR22725. 3.3. Mode Synthesizing Atomic Force
Microscopy (MSAFM) and Hybrid Photonic
Force Microscopy (HPFM): Chemical
Compositiont With the results reported thus far on plant cell imaging
with a resolution of ~5 nm, and on cell wall Young’s modulus
and plastic index with typical values in the ranges 0.3–5 GPa and
0–0.5, respectively, quantitative biomass characterization is only
at its debut. Considering the potential of the emerging AFM-
based configurations of MSAFM and HPFM for high-resolution
physical and chemical studies of the subsurface domains of the
plant cells, we envision that novel and targeted studies based on
innovative AFM modalities will continue to play an important
role in establishing the basic characteristics of the plant cells as
well as contribute to streamlining of biofuel production. The cellular ultrastructure was further revealed with HPFM with
the addition of modulated infrared light at λ = 10,200 nm at
ωQCL → ω− = 26 kHz. Cellulose and lignin both absorb light at
λ = 10,200 nm, but at different absorption intensities, resulting
in contrasted signal (Figure 4D). The surface and subsurface
structures disclosed remarkable detail, especially in the cellulose-
rich region and the small grain-like structures outlined in black
were interpreted to be cellulosic microfibril aggregates (globules),
which are known to be located in the secondary cell wall (Salmén,
2004). MSAFM was also used to study the effect of holopulping
treatment on holocellulose poplar sections. It was reported that
the holopulping process, intended to oxidatively remove the
lignin network in the cell, appeared to affect mostly the middle
lamella as well as the cell corner regions (Tetard et al., 2011). Asina, F., Brzonova, I., Voeller, K., Kozliak, E., Kubatova, A., Yao, B., et al. (2016).
Biodegradation of lignin by fungi, bacteria and laccases. Bioresour. Technol.
220, 414–424. doi:10.1016/j.biortech.2016.08.016 Al-Zuhair, S., Abualreesh, M., Ahmed, K., and Razak, A. A. (2015). Enzymatic
delignification of biomass for enhanced fermentable sugars production. Energy
Technol. 3, 121–127. doi:10.1002/ente.201402138 AUTHOR CONTRIBUTIONS All authors contributed to the manuscript. Al-Zuhair, S., Abualreesh, M., Ahmed, K., and Razak, A. A. (2015). Enzymatic
delignification of biomass for enhanced fermentable sugars production. Energy
Technol. 3, 121–127. doi:10.1002/ente.201402138
Asina, F., Brzonova, I., Voeller, K., Kozliak, E., Kubatova, A., Yao, B., et al. (2016).
Biodegradation of lignin by fungi, bacteria and laccases. Bioresour. Technol.
220, 414–424. doi:10.1016/j.biortech.2016.08.016 Al-Zuhair, S., Abualreesh, M., Ahmed, K., and Razak, A. A. (2015). Enzymatic
delignification of biomass for enhanced fermentable sugars production. Energy
Technol. 3, 121–127. doi:10.1002/ente.201402138 REFERENCES Plant Methods 10, 1. doi:10.1186/1746-4811-10-1 Burgert, I., and Keplinger, T. (2013). Plant micro- and nanomechanics: exper-
imental techniques for plant cell-wall analysis. J. Exp. Bot. 64, 4635–4649. doi:10.1093/jxb/ert255 Butt, H., Cappella, B., and Kappl, M. (2005). Force measurements with the atomic
force microscope: technique, interpretation and applications. Surf. Sci. Rep. 59,
1–152. doi:10.1016/j.surfrep.2005.08.003 Kirby, A. R. (2011). Atomic force microscopy of plant cell walls. Methods Mol Biol
715, 169–178. doi:10.1007/978-1-61779-008-9_12 Kirby, A., Gunning, A., Waldron, K., Morris, V., and Ng, A. (1996). Visualization
of plant cell walls by atomic force microscopy. Biophys J 70, 1138–1143. doi:10.1016/S0006-3495(96)79708-4 Cadoche, L., and Lopez, G. (1989). Assessment of size-reduction as a preliminary
step in the production of ethanol from lignocellulosic wastes. Biol. Wastes 30,
153–157. doi:10.1016/0269-7483(89)90069-4 Konnerth, J., Harper, D., Lee, S.-H., Rials, T. G., and Gindl, W. (2008). Adhesive
penetration of wood cell walls investigated by scanning thermal microscopy
(SThM). Holzforschung 62, 91–98. doi:10.1515/HF.2008.014 Carlos Martinez-Patino, J., Ruiz, E., Romero, I., Cara, C., Carlos Lopez-Linares, J.,
and Castro, E. (2017). Combined acid/alkaline-peroxide pretreatment of olive
tree biomass for bioethanol production. Bioresour. Technol. 239, 326–335. doi:10.1016/j.biortech.2017.04.102 f Lynd, L. R., Laser, M. S., Bransby, D., Dale, B. E., Davison, B., Hamilton, R., et al. (2008). How biotech can transform biofuels. Nat. Biotechnol. 26, 169–172. doi:10.1038/nbt0208-169 Chen, X., Li, H., Sun, S., Cao, X., and Sun, R. (2016). Effect of hydrothermal
pretreatment on the structural changes of alkaline ethanol lignin from wheat
straw. Sci. Rep. 6, 39354. doi:10.1038/srep39354 f Martin, Y., Williams, C., and Wickramasinghe, H. (1987). Atomic force microscope
force mapping and profiling on a sub 100-a scale. J. Appl. Phys. 61, 4723–4729. doi:10.1063/1.338807 Derjaguin, B., Muller, V., and Toporov, Y. (1975). Effect of contact defor-
mations on adhesion of particles. J. Colloid Interface Sci. 53, 314–326. doi:10.1016/0021-9797(75)90018-1 Medeiros, R. G., Silva, L. P., Azevedo, R. B., Silva, F. G. Jr., Ximenes, F., and Filho, E. (2007). The use of atomic force microscopy as a tool to study the effect of a
xylanase from Humicola grisea var. thermoidea in kraft pulp bleaching. Enzyme
Microb. Technol. 40, 723–731. doi:10.1016/j.enzmictec.2006.06.004 h Durkovic, J., Kardosova, M., and Lagana, R. (2014). Imaging and measurement
of nanomechanical properties within primary xylem cell walls of broadleaves. Bio-protocol 4, 1360. doi:10.21769/BioProtoc.1360 Mohapatra, S., Mishra, C., Behera, S. S., and Thatoi, H. (2017). Application of
pretreatment, fermentation and molecular techniques for enhancing bioetha-
nol production from grass biomass – a review. Renew. REFERENCES Adamcik, J., Berquand, A., and Mezzenga, R. (2011). Single-step direct
measurement of amyloid fibrils stiffness by peak force quantitative nanome-
chanical atomic force microscopy. Appl. Phys. Lett. 98, 193701. doi:10.1063/
1.3589369 March 2018 | Volume 6 | Article 11 Frontiers in Energy Research | www.frontiersin.org 7 AFM on Plant Cell Walls? Charrier et al. Auxenfans, T., Cronier, D., Chabbert, B., and Paes, G. (2017). Understanding the
structural and chemical changes of plant biomass following steam explosion
pretreatment. Biotechnol. Biofuels 10, 36. doi:10.1186/s13068-017-0718-z Hanley, S., Revol, J., Godbout, L., and Gray, D. (1997). Atomic force microscopy
and transmission electron microscopy of cellulose from Micrasterias denticu-
lata; evidence for a chiral helical microfibril twist. Cellulose 4, 209–220. doi:10
.1023/A:1018483722417 Baker, A., Helbert, W., Sugiyama, J., and Miles, M. (2000). New insight into
cellulose structure by atomic force microscopy shows the i-alpha crystal
phase at near-atomic resolution. Biophys. J. 79, 1139–1145. doi:10.1016/
S0006-3495(00)76367-3 Healey, A. L., Lee, D. J., Furtado, A., Simmons, B. A., and Henry, R. J. (2015). Efficient
eucalypt cell wall deconstruction and conversion for sustainable lignocellu
losic biofuels. Front. Bioeng. Biotechnol. 3:190. doi:10.3389/fbioe.2015.00190 Beecher, J. F., Hunt, C. G., and Zhu, J. Y. (2009). “Tools for the Characterization
of Biomass at the Nanometer Scale,” in The Nanoscience and Technology of
Renewable Biomaterials, eds L. A. Lucia, and O. J. Rojas (Wiley-Blackwell),
61–90. b
Herruzo, E. T., Perrino, A. P., and Garcia, R. (2014). Fast nanomechanical spect
copy of soft matter. Nat. Commun. 5, 3126. doi:10.1038/ncomms4126 Herruzo, E. T., Perrino, A. P., and Garcia, R. (2014). Fast nanomechanical spectros-
copy of soft matter. Nat. Commun. 5, 3126. doi:10.1038/ncomms4126 opy of soft matter. Nat. Commun. 5, 3126. doi:10.1038/ncomms412 t
Heu, C., Berquand, A., Elie-Caille, C., and Nicod, L. (2012). Glyphosate-induced
stiffening of hacat keratinocytes, a peak force tapping study on living cells. J. Struct. Biol. 178, 1–7. doi:10.1016/j.jsb.2012.02.007 Binnig, G., Quate, C. F., and Gerber, C. (1986). Atomic force microscope. Phys. Rev. Lett. 56, 930–933. doi:10.1103/PhysRevLett.56.930 Johnson, K., Kendall, K., and Roberts, A. (1971). Surface energy and contact of
elastic solids. Proc. R. Soc. Lond. Ser. A Math. Phys. Sci. 324, 301. doi:10.1098/
rspa.1971.0141 Bulichev, S., and Alekhin, V. (1987). Method of kinetic hardness and micro
hardness in testing impression by an indentor. Ind. Lab. 53, 1091–1096. Keplinger, T., Konnerth, J., Aguie-Beghin, V., Rueggeberg, M., Gierlinger, N., and
Burgert, I. (2014). A zoom into the nanoscale texture of secondary cell walls. REFERENCES Sustain. Energ. Rev. 78,
1007–1032. doi:10.1016/j.rser.2017.05.026 Erdocia, X., Prado, R., Angeles Corcuera, M., and Labidi, J. (2014). Effect of
different organosolv treatments on the structure and properties of olive tree
pruning lignin. J. Ind. Eng. Chem. 20, 1103–1108. doi:10.1016/j.jiec.2013.06.048 Fang, W., Yang, S., Wang, X.-L., Yuan, T.-Q., and Sun, R.-C. (2017). Manufacture
and application of lignin-based carbon fibers (LCFs) and lignin-based carbon Fang, W., Yang, S., Wang, X.-L., Yuan, T.-Q., and Sun, R.-C. (2017). Manufacture
and application of lignin-based carbon fibers (LCFs) and lignin-based carbon
nanofibers (LCNFs). Green Chem. 19, 1794–1827. doi:10.1039/C6GC03206K Nanda, S., Maley, J., Kozinski, J. A., and Dalai, A. K. (2015). Physico-chemical
evolution in lignocellulosic feedstocks during hydrothermal pretreatment
and delignification. J. Biobased Mater. bioenergy 9, 295–308. doi:10.1166/
jbmb.2015.1529 and application of lignin-based carbon fibers (LCFs) and lignin-based carbon
nanofibers (LCNFs). Green Chem. 19, 1794–1827. doi:10.1039/C6GC03206K Farahi, R., Charrier, A., Tolbert, A., Lereu, A., Ragauskas, A., Davison, B., et al. (2017). Plasticity, elasticity, and adhesion energy of plant cell walls: nanometrol-
ogy of lignin loss using atomic force microscopy. Sci. Rep. 7, 152. doi:10.1038/
s41598-017-00234-4 Peaucelle, A. (2014). Afm-based mapping of the elastic properties of cell
walls: at tissue, cellular, and subcellular resolutions. J Vis Exp 24, e51317. doi:10.3791/51317 Fernandes, A. N., Chen, X., Scotchford, C. A., Walker, J., Wells, D. M., Roberts, C. J., et al. (2012). Mechanical properties of epidermal cells of whole living roots
of Arabidopsis thaliana: an atomic force microscopy study. Phys. Rev. E Stat. Nonlin. Soft. Matter. Phys. 85, 1. doi:10.1103/PhysRevE.85.021916 Pharr, G., Oliver, W., and Brotzen, F. (1992). On the generality of the relationship
among contact stiffness, contact area, and elastic-modulus during indentation. J. Mater. Res. 7, 613–617. doi:10.1557/JMR.1992.0613 Pilate, G., Guiney, E., Holt, K., Petit-Conil, M., Lapierre, C., Leple, J., et al. (2002). Field and pulping performances of transgenic trees with altered lignification. Nat. Biotechnol. 20, 607–612. doi:10.1038/nbt0602-607 t
Fratzl, P., and Weinkamer, R. (2007). Nature’s hierarchical materials. Prog. Mater. Sci. 52, 1263–1334. doi:10.1016/j.pmatsci.2007.06.001 Radmacher, M. (1997). Measuring the elastic properties of biological samples with
the afm. IEEE Eng. Med. Biol. Mag. 16, 47–57. doi:10.1109/51.582176 Garcia, R., Magerle, R., and Perez, R. (2007). Nanoscale compositional mapping
with gentle forces. Nat. Mater. 6, 405–411. doi:10.1038/nmat1925 Garcia, R., and Perez, R. (2002). Dynamic atomic force microscopy methods. Surf. Sci. Rep. 47, 197–301. doi:10.1016/S0167-5729(02)00077-8
Gibson, L. J. (2012). The hierarchical structure and mechanics of plant materials. J. R. Soc. Interface 9, 2749–2766. REFERENCES Longitudinal hardness and
young’s modulus of spruce tracheid secondary walls using nanoindentation
technique. Wood Sci. Technol. 31, 131–141. doi:10.1007/BF00705928 Singh, R., Shukla, A., Tiwari, S., and Srivastava, M. (2014). A review on delignifica-
tion of lignocellulosic biomass for enhancement of ethanol production poten-
tial. Renewable Sustain. Energy Rev. 32, 713–728. doi:10.1016/j.rser.2014.01.051 Yakubov, G. E., Bonilla, M. R., Chen, H., Doblin, M. S., Bacic, A., Gidley, M. J.,
et al. (2016). Mapping nano-scale mechanical heterogeneity of primary plant
cell walls. J. Exp. Bot. 67, 2799–2816. doi:10.1093/jxb/erw117 Youssefian, S., Jakes, J. E., and Rahbar, N. (2017). Variation of nanostructures,
molecular interactions, and anisotropic elastic moduli of lignocellulosic cell
walls with moisture. Sci. Rep. 7, 2054. doi:10.1038/s41598-017-02288-w Singh, S., Cheng, G., Sathitsuksanoh, N., Wu, D., Varanasi, P., George, A., et al. (2015). Comparison of different biomass pretreatment techniques and their
impact on chemistry and structure. Front. Energy Res. 2:62. doi:10.3389/
fenrg.2014.00062 Zdunek, A., and Kurenda, A. (2013). Determination of the elastic properties
of tomato fruit cells with an atomic force microscope. Sensors (Basel) 13,
12175–12191. doi:10.3390/s130912175 Spitzner, E.-C., Roeper, S., Zerson, M., Bernstein, A., and Magerle, R. (2015). Nanoscale swelling heterogeneities in type I collagen fibrils. ACS Nano 9,
5683–5694. doi:10.1021/nn503637q Zhang, M., Chen, G., Kumar, R., and Xu, B. (2013a). Mapping out the structural
changes of natural and pretreated plant cell wall surfaces by atomic force
microscopy single molecular recognition imaging. Biotechnol. Biofuels 6, 147. doi:10.1186/1754-6834-6-147 fi Tetard, L., Passian, A., Farahi, R. H., Davison, B. H., Jung, S., Ragauskas, A. J.,
et al. (2011). Nanometrology of delignified populus using mode synthesizing
atomic force microscopy. Nanotechnology 22, 465702. doi:10.1088/0957-
4484/22/46/465702 Zhang, M., Wang, B., and Xu, B. (2013b). Measurements of single molecular affinity
interactions between carbohydrate-binding modules and crystalline cellulose
fibrils. Phys. Chem. Chem. Phys. 15, 6508–6515. doi:10.1039/c3cp51072g Tetard, L., Passian, A., Farahi, R. H., Thundat, T., and Davison, B. H. (2015). Opto-
nanomechanical spectroscopic material characterization. Nat. Nanotechnol. 10,
870–877. doi:10.1038/nnano.2015.168 Zhang, T., Vavylonis, D., Durachko, D. M., and Cosgrove, D. J. (2017). Nanoscale
movements of cellulose microfibrils in primary cell walls. Nat. Plants 3, 17056. doi:10.1038/nplants.2017.56 Tetard, L., Passian, A., and Thundat, T. (2010). New modes for subsurface atomic
force microscopy through nanomechanical coupling. Nat. Nanotechnol. 5,
105–109. doi:10.1038/nnano.2009.454 Zhong, Q., Inniss, D., Kjoller, K., and Elings, V. (1993). Fractured polymer silica
fiber surface studied by tapping mode atomic-force microscopy. Surf. Sci. 290,
L688–L692. REFERENCES doi:10.1098/rsif.2012.0341 Garcia, R., and Perez, R. (2002). Dynamic atomic force microscopy methods. Surf. Sci. Rep. 47, 197–301. doi:10.1016/S0167-5729(02)00077-8 h Radmacher, M., Fritz, M., Kacher, C., Cleveland, J., and Hansma, P. (1996). Measuring the viscoelastic properties of human platelets with the atomic force
microscope. Biophys. J. 70, 556–567. doi:10.1016/S0006-3495(96)79602-9 Sci. Rep. 47, 197–301. doi:10.1016/S0167-5729(02)00077-8
Gibson, L. J. (2012). The hierarchical structure and mechanics of plant materials. J R Soc Interface 9 2749 2766 doi:10 1098/rsif 2012 0341 p
(
)
Gibson, L. J. (2012). The hierarchical structure and mechanics of plant materials. J. R. Soc. Interface 9, 2749–2766. doi:10.1098/rsif.2012.0341 Gibson, L. J. (2012). The hierarchical structure and mechanics of plant materials. J. R. Soc. Interface 9, 2749–2766. doi:10.1098/rsif.2012.0341 J (
)h
p
J. R. Soc. Interface 9, 2749–2766. doi:10.1098/rsif.2012.0341 Radotic, K., Roduit, C., Simonovic, J., Hornitschek, P., Fankhauser, C., Mutavdzic, D.,
et al. (2012). Atomic force microscopy stiffness tomography on living
Arabidopsis thaliana cells reveals the mechanical properties of surface and deep
cell-wall layers during growth. Biophys. J. 103, 386–394. doi:10.1016/j.bpj.2012. 06.046 f
Gindl, W., and Gupta, H. (2002). Cell-wall hardness and young’s modulus of
melamine-modified spruce wood by nano-indentation. Compos. Part A Appl. Sci. Manuf. 33, 1141–1145. doi:10.1016/S1359-835X(02)00080-5 Gindl, W., Gupta, H., Schoberl, T., Lichtenegger, H., and Fratzl, P. (2004). Mechanical
properties of spruce wood cell walls by nanoindentation. Appl. Phys. A Mater. Sci. Process. 79, 2069–2073. doi:10.1007/s00339-004-2864-y hi Salmén, L. (2004). Micromechanical understanding of the cell-wall structure. C. R. Biol. 327, 873–880. doi:10.1016/j.crvi.2004.03.010 p
p
p
y
pp
y
Sci. Process. 79, 2069–2073. doi:10.1007/s00339-004-2864-y
Gindl, W., and Schoberl, T. (2004). The significance of the elastic modulus of wood
cell walls obtained from nanoindentation measurements. Compos. Part A
Appl. Sci. Manuf. 35, 1345–1349. doi:10.1016/j.compositesa.2004.04.002 Gindl, W., and Schoberl, T. (2004). The significance of the elastic modulus of wood
cell walls obtained from nanoindentation measurements. Compos. Part A
Appl. Sci. Manuf. 35, 1345–1349. doi:10.1016/j.compositesa.2004.04.002 Salvadori, M. R., Oller Nascimento, C. A., and Correa, B. (2014). Nickel oxide
nanoparticles film produced by dead biomass of filamentous fungus. Sci. Rep. 4, 6404. doi:10.1038/srep06404 March 2018 | Volume 6 | Article 11 Frontiers in Energy Research | www.frontiersin.org 8 AFM on Plant Cell Walls? Charrier et al. Sannigrahi, P., Ragauskas, A. J., and Tuskan, G. A. (2010). Poplar as a feedstock for
biofuels: a review of compositional characteristics. Biofuels Bioprod. Biorefin. 4,
209–226. doi:10.1002/bbb.206 Wimmer, R., Lucas, B., Tsui, T., and Oliver, W. (1997). Frontiers in Energy Research | www.frontiersin.org REFERENCES doi:10.1016/0039-6028(93)90582-5 Tetard, L., Passian, A., Venmar, K. T., Lynch, R. M., Voy, B. H., Shekhawat, G.,
et al. (2008). Imaging nanoparticles in cells by nanomechanical holography. Nat. Nanotechnol. 3, 501–505. doi:10.1038/nnano.2008.162 Zickler, G., Schoberl, T., and Paris, O. (2006). Mechanical properties of
pyrolysed wood: a nanoindentation study. Philos. Mag. 86, 1373–1386. doi:10.1080/14786430500431390 Tian, D., Chandra, R. P., Lee, J.-S., Lu, C., and Saddler, J. N. (2017). A comparison
of various lignin-extraction methods to enhance the accessibility and ease of
enzymatic hydrolysis of the cellulosic component of steam-pretreated poplar. Biotechnol. Biofuels 10, 157. doi:10.1186/s13068-017-0846-5 Conflict of Interest Statement: The authors declare that the research was con-
ducted in the absence of any commercial or financial relationships that could be
construed as a potential conflict of interest. Tolbert, A., and Ragauskas, A. J. (2017). Advances in understanding the surface
chemistry of lignocellulosic biomass via time-of-flight secondary ion mass
spectrometry. Energy Sci. Eng. 5, 5–20. doi:10.1002/ese3.144 Torode, T. A., O’Neill, R., Marcus, S. E., Cornuault, V., Pose, S., Lauder, R. P.,
et al. (2018). Branched pectic galactan in phloem-sieve-element cell walls:
Implications for cell mechanics. Plant Physiol 176, 1547–1558. doi:10.1104/
pp.17.01568 The reviewer, MR, and handling editor declared their shared affiliation. The reviewer, MR, and handling editor declared their shared affiliation. Copyright © 2018 Charrier, Lereu, Farahi, Davison and Passian. This is an open-
access article distributed under the terms of the Creative Commons Attribution
License (CC BY). The use, distribution or reproduction in other forums is permitted,
provided the original author(s) and the copyright owner are credited and that the
original publication in this journal is cited, in accordance with accepted academic
practice. No use, distribution or reproduction is permitted which does not comply
with these terms. Wagner, L., Bader, T. K., and de Borst, K. (2014). Nanoindentation of wood cell
walls: effects of sample preparation and indentation protocol. J. Sci. Mater. 49,
94–102. doi:10.1007/s10853-013-7680-3 Wimmer, R., and Lucas, B. (1997). Comparing mechanical properties of second-
ary wall and cell corner middle lamella in spruce wood. IAWA J. 18, 77–88. doi:10.1163/22941932-90001463 March 2018 | Volume 6 | Article 11 Frontiers in Energy Research | www.frontiersin.org 9
|
https://openalex.org/W2934649944
|
https://ms.copernicus.org/articles/10/119/2019/ms-10-119-2019.pdf
|
English
| null |
Laparoscope arm automatic positioning for robot-assisted surgery based on reinforcement learning
|
Mechanical sciences
| 2,019
|
cc-by
| 8,629
|
Laparoscope arm automatic positioning for
robot-assisted surgery based on reinforcement learning Lingtao Yu1, Xiaoyan Yu1, Xiao Chen1, and Fengfeng Zhang2
1College of Mechanical and Electrical Engineering, Harbin Engineering University, Harbin, 150001, China
2School of Mechanical and Electrical Engineering, Soochow University, Suzhou, 215021, China
Correspondence: Xiaoyan Yu (xiaoyanheu10@gmail.com)
Received: 5 July 2018 – Revised: 12 March 2019 – Accepted: 20 March 2019 – Published: 3 April 2019 Abstract. Compared with the traditional laparoscopic surgery, the preoperative planning of robot-assisted la-
paroscopic surgery is more complex and essential. Through the analysis of the surgical procedures and surgical
environment, the laparoscope arm preoperative planning algorithm based on the artificial pneumoperitoneum
model, lesion parametrization model is proposed, which ensures that the laparoscope arm satisfies both the
distance principle and the direction principle. The algorithm is divided into two parts, including the optimum
incision and the optimum angle of laparoscope entry, which makes the laparoscope provide a reasonable ini-
tial visual field. A set of parameters based on the actual situation is given to illustrate the algorithm flow in
detail. The preoperative planning algorithm offers significant improvements in planning time and quality for
robot-assisted laparoscopic surgery. The improved method which combines the preoperative planning algorithm
with deep deterministic policy gradient algorithm is applied to laparoscope arm automatic positioning for the
robot-assisted laparoscopic surgery. It takes a fixed-point position and lesion parameters as input, and outputs
the optimum incision, the optimum angle and motor movements without kinematics. The proposed algorithm
is verified through simulations with a virtual environment built by pyglet. The results validate the correctness,
feasibility, and robustness of this approach. Published by Copernicus Publications. Mech. Sci., 10, 119–131, 2019
https://doi.org/10.5194/ms-10-119-2019
© Author(s) 2019. This work is distributed under
the Creative Commons Attribution 4.0 License. Mech. Sci., 10, 119–131, 2019
https://doi.org/10.5194/ms-10-119-2019
© Author(s) 2019. This work is distributed under
the Creative Commons Attribution 4.0 License. Laparoscope arm automatic positioning for
robot-assisted surgery based on reinforcement learning L. Yu et al.: Laparoscope arm automatic positioning (2010) proposed a new method of
port placement for laparoscopic radical prostatectomy, which
used a nautical inclinometer and a homemade triangle mold
(Cestari et al., 2010). The heuristic method based on the surgeon experience is
convenient and practical for the surgeon, so it is widely used
in clinical practice. However, this method is related to the
surgeon’s operating habits and requires extensive surgical ex-
perience. More importantly, the advantages of the surgical
robot system are not fully developed. Hayashibe et al. (2005) developed the simulation system
for preoperative planning of abdominal surgery. The core of
the simulation system was kinematics and haptics; the ef-
fectiveness of preoperative planning was validated by the
surgeon’s evaluation (Hayashibe et al., 2005). Hayashibe et
al. (2006) developed a new simulation system with volume
rendering of medical images and automatic positioning by
kinematics (Hayashibe et al., 2006). Sun et al. (2007) devel-
oped a simulator of the da Vinci system, which was mainly
used for surgeon training. Its primary functions were the sim-
ulation of port placement and the practice of simple surgical
operations (Sun et al., 2007). Bauernschmitt et al. (2007) de-
veloped a simulator for port placement and enhanced guid-
ance in robot-assisted heart surgery. The simulator was com-
pleted off-line, the simulation model is established by using
the patient’s computed tomography (CT) images to get the
best ports position. Through this system, preoperative plan-
ning was optimized, the operation time was reduced, and op-
eration quality was improved (Bauernschmitt et al., 2007). Konietschke et al. (2011) developed a simulator of the DLR
MiroSurge system, which used the VR-Map device to estab-
lish the simulator quickly. Its primary functions were pre-
operative optimization and intraoperative simulation (Koni-
etschke et al., 2011). Compared with the former two methods, the method based
on multi-objective optimization algorithm is more scientific. More importantly, in addition to the surgeon experience, the
robot’s characteristics are also taken into account, so the pre-
operative planning is more conducive to the operation. In general, after obtaining the preoperative planning by
the above method, the joint variables of the manipulator are
obtained by inverse kinematics. At present, the telecentric
fixed-point positioning mechanism of the surgical robot sys-
tem is mostly an undriven mechanism, which needs to be
manually adjusted to the target position. L. Yu et al.: Laparoscope arm automatic positioning L. Yu et al.: Laparoscope arm automatic positioning L. Yu et al.: Laparoscope arm automatic positioning Sun and Yeung (2007) proposed the selection of op-
timal port placement and the determination of optimal
robot attitude based on multi-objective optimization. This
method used two performance indices, the global isotropy
index (GII) and the efficiency index (EI). Through the inter-
action of these two indicators, the flexibility and operability
of the robot were improved, and the workspace and visual
space were also increased (Sun and Yeung, 2007). Azimian
et al. (2010) proposed the preoperative planning method for
robot-assisted minimally invasive CABG. This method used
sequential quadratic programming to implement the opti-
mization of kinematic and geometric requirements. In the
optimization process, individualized preoperative planning
can be achieved taking into account the surgeon’s experi-
ence (Azimian et al., 2010). Ma et al. (2014) proposed the
preoperative positioning method, which was mainly aimed at
the collision problem of the multi-arm system. It used the
maximum distance index to achieve collision-free optimal
preoperative positioning (Ma et al., 2014). Yu et al. (2014)
proposed the preoperative positioning method, which was
mainly aimed at cooperative cooperation between two instru-
ment arms. It used the percentage of collaboration workspace
to achieve the optimal cooperation between two manipula-
tors (Yu et al., 2014). Wang et al. (2016) proposed a preoper-
ative planning algorithm for robot-assisted minimally inva-
sive CABG. This algorithm used two performance indices,
isotropy index based on CV (IICV) and index of instrument
collaboration space (IICS), to implement the optimal port
placement selection and the manipulator poses determination
(Wang et al., 2016). zli and Fingerhut (2004) proposed recommendations of tro-
car placement for laparoscopic surgery. The abdominal cav-
ity is divided into six parts according to the operation area,
and recommendations are given according to different oper-
ations and patient posture characteristics (Ferzli and Finger-
hut, 2004). Pick et al. (2014) proposed an anatomic guide
of port placement for laparoscopic radical prostatectomy,
which was performed on the da Vinci robot-assisted surgi-
cal system. Compared to traditional port placement, the pu-
bic bone was used as optimal landmark (Pick et al., 2004). Badani et al. (2008) proposed a novel technique of port place-
ment for robotic renal surgery, which aimed to maximize the
range of motion and eliminate external collisions (Badani et
al., 2008). Cestari et al. Mech. Sci., 10, 119–131, 2019 1
Introduction reasonable preoperative planning can significantly reduce the
operation time; otherwise, it may increase surgical risks. With the development of robotic technology and application
of minimally invasive surgery (MIS), the laparoscopic MIS
robotic system has been widely used in surgical specialties,
such as urology (prostate, bladder and kidney cancer), gy-
necology (hysterectomy and myomectomy). Compared with
traditional laparoscopic surgery, robot-assisted laparoscopic
surgery displays high-definition, 3-D image of the lesion to
the surgeon via the console and allows the surgeon to per-
form complex operations by manipulating the master con-
trols. Robot-assisted laparoscopic surgery is more precision,
flexibility, and controllable than conventional techniques, so
it has become the research hotspot in recent years. For MIS robotic system preoperative planning, scholars
have proposed many different methods, which are divided
into three parts: (1) A heuristic method based on surgeon ex-
perience. (2) A method based on the virtual surgical environ-
ment. (3) A method based on multi-objective optimization
algorithm. Hanna et al. (1997a) investigated the impact of port place-
ment on endoscopic manipulations, especially knotting. The
optimal azimuth and elevation angles were obtained by com-
paring the execution time and performance quality score
of tying a surgeon’s knot (Hanna et al., 1997a). Austad et
al. (2001) completed the coronary artery bypass grafting pro-
cedures on pigs using the Zeus robot-assisted surgical sys-
tem. The Zeus system configurations, like port placement
and pigs’ position, were set based on recommendations from
hospitals and surgeon experience (Austad et al., 2001). Fer- Although robot-assisted surgery has many advantages over
traditional surgery, there are also some thorny problems, such
as control switching between master controls and robotic
arms, real-time synchronization of master-slave position and
attitude, MIS robotic system preoperative planning. Besides, 120 L. Yu et al.: Laparoscope arm automatic positioning Due to errors of
manual adjustment and mechanical kinematics parameters,
the actual preoperative planning is not the optimal solution
previously determined. Therefore, it is necessary to use a new
method to complete preoperative planning instead of manual
configuration. Traditional manipulator control is to calculate joint vari-
ables by inverse kinematics of a given target position. At
present, its trend has turned to the end-to-end solution. In
other words, the controller learns diverse strategies directly
from sensors data, rather than relying on fixed strategies such
as kinematics (James and Johns, 2016; Otte et al., 2016;
Phaniteja et al., 2017; Gu et al., 2017; Mohammadi et al.,
2018). James and Johns (2016) proposed a method that took
images as its input and outputs motor movements and target The method based on the virtual surgical environment vi-
sualizes the port placement and verifies the effect in advance. Compared with the former method, this method simplifies
the steps of port placement and reduces the time required. However, this method also requires surgeons with extensive
surgical experience, and due to the lack of analysis of sur-
gical robot performance and finite attempts, it is difficult to
obtain optimized preoperative planning. www.mech-sci.net/10/119/2019/ L. Yu et al.: Laparoscope arm automatic positioning 121 Figure 1. The schematic diagram of surgical incisions. position. Thus, the control of the 7-DOF robot arm can be re-
alized in a virtual environment without any prior knowledge
(James and Johns, 2016). The telecentric fixed-point posi-
tioning mechanism is a redundant mechanism; an accurate
kinematic inverse solution can only be obtained under ap-
propriate constraints. In order to improve the effect of preop-
erative planning, it is necessary to explore a new method to
tackle the problems caused by previous methods. This paper proposes a laparoscope arm preoperative plan-
ning algorithm, which is based on the lesion parametrization
model and evaluation indexes. Besides, an improved method
based on reinforcement learning algorithm is proposed to
achieve preoperative laparoscope arm automatic positioning. More importantly, it is a crucial step towards the automation
of robot-assisted laparoscopic surgery. Figure 1. The schematic diagram of surgical incisions. The rest of the paper is organized as follows. Section 2 in-
troduces surgical procedures and MIS robotic system. The la-
paroscope arm preoperative planning algorithm is introduced
in Sect. 3. The improved DDPG algorithm is introduced in
Sect. 4. The simulation results are presented in Sect. 5. 2.1
The MIS procedures The common MIS has three steps: (1) According to the actual
surgical needs, a surgeon makes several small incisions (usu-
ally 5–15 mm) and inserts a thin tube called trocar. The tro-
car is deployed as a means of introduction for laparoscope or
laparoscopic instruments, like scissors and graspers, to pro-
vide an access port during surgery. (2) Creation of a pneu-
moperitoneum by inflating the abdomen with carbon dioxide
to make a separation between organs and increase the operat-
ing space of surgical instruments. (3) The surgeon views the
magnified image of the patient’s internal organs provided by
laparoscope on a video monitor. Using different instruments,
the surgeon performs a series of surgical operations in the
pneumoperitoneum. L. Yu et al.: Laparoscope arm automatic positioning Dis-
cussion and conclusion are given in Sects. 6 and 7, respec-
tively. of one laparoscope arm and two instrument arms. Laparo-
scope arm is equipped with a laparoscope, and instrument
arms are equipped with different laparoscopic instruments. Laparoscope arm and instrument arms are located on both
sides of the operating bed. A depth camera is installed above
the operating bed for acquiring the position of the incisions
and robotic arms, as shown in Fig. 2. 2
Robot-assisted surgery The three arms have the same mechanical structure. Each
arm is divided into three parts, the telecentric fixed-point po-
sitioning mechanism, the remote center of motion mecha-
nism and the end effector, as shown in Fig. 3. The first part
adjusts the spatial position of telecentric fixed-point by three
revolving joints and one linear joint. The second part adjusts
the position and posture of the end effector by the master ma-
nipulator operated by a surgeon; at its end, there is a versatile
quick-change mechanism for end effectors installation. 3
Laparoscope arm preoperative planning One of the critical issues for MIS is preoperative planning,
including preparation for interventions and decision about
the optimum surgical incisions. Currently, the surgeon of-
ten uses trial-and-error method or experience-based to com-
plete preoperative planning, which may not meet the require-
ments of the optimum incisions. Therefore, it is necessary
to use preoperative planning algorithm instead of the previ-
ous method. The preoperative planning includes laparoscope
arm and instrument arms preoperative planning. This paper
studies the former, including the optimum incision and the
optimum angle of laparoscope entry. This paper takes laparoscopic cholecystectomy (LC) as
an example. The surgeon makes three incisions and inserts
trocar. In LC, it is always with the patient in a supine po-
sition. Three incisions are arranged in an isosceles triangle
for better operating space, as shown in Fig. 1. A laparoscope
is placed through a trocar, and specialized instruments are
placed through other trocars. By operating the laparoscope
and instruments, the surgeon delicately separates the gall-
bladder from its attachments to the liver and the bile duct
and then removes it through one incision. 2
Robot-assisted surgery www.mech-sci.net/10/119/2019/ Mech. Sci., 10, 119–131, 2019 2.2
Layout design of MIS robotic system The mathematical model of pneumoperitoneum is estab-
lished before preoperative planning. The shape of artificial
pneumoperitoneum is approximately ellipsoid (Mulier et al., The MIS robotic system includes a master-slave manipulator
system and a depth camera. The slave manipulator consists www.mech-sci.net/10/119/2019/ Mech. Sci., 10, 119–131, 2019 L. Yu et al.: Laparoscope arm automatic positioning L. Yu et al.: Laparoscope arm automatic positioning L. Yu et al.: Laparoscope arm automatic positioning 122 L. Yu et al.: Laparoscope arm automatic positioning
Figure 4. The coordinate frame and pneumoperitoneum model. Figure 2. The MIS robotic system. Figure 4. The coordinate frame and pneumoperitoneum model. as shown in Fig. 4. x2
a2p
+ y2
b2p
+ z2
c2p
= 1
(1) (1) During actual operation, the model parameters (ap, bp, cp)
are determined by the medical image and gas insufflation
volume. Suppose an adult’s chest width is 3.15 dm, chest
thickness is 2.45 dm, and chest length is 2.9 dm. The cor-
responding parameters in Fig. 6 are ap = 1.55, bp = 1.45,
h = 1.2 dm. Chen suggested that the gas insufflation volume
is about 3L (Chen, 1999). According to Eq. (2), calculate
cp = 2.27 dm. Figure 2. The MIS robotic system. Figure 3. The structure of the robotic arm. V =
cpZ
h
πapbp(1 −z2
c2p
)dz = πapbp(2
3cp −h + 1
3
h3
c2p
)
(2) (2) 3.3
The preoperative planning algorithm framework Through the study of the mathematical model of artificial
pneumoperitoneum, lesion parametrization model and pre-
operative planning principles, the laparoscope arm preopera-
tive planning algorithm is proposed, as shown in Fig. 6, that
includes three stages: data processing and modeling, opti-
mum incision determination and optimum angle determina-
tion. Figure 6. Flow chart of the laparoscope arm preoperative planning
algorithm. In the first stage, obtain patient information from the med-
ical images, and then establish the mathematical model of ar-
tificial pneumoperitoneum, and lastly determine the location
and lesion parametrization model. This stage is the basis of
the entire algorithm, and also the most time-consuming stage. In the second stage, all allowable surgical incisions are ob-
tained from the first stage, and then the candidate incisions
are determined according to the two principles. The candi-
date base positions are obtained by the candidate incisions. According to the actual situation of the operating room, se-
lect one of the positions as the base position. Combine can-
didate incisions and the base position to determine the opti-
mum incision. In the first stage, obtain patient information from the med-
ical images, and then establish the mathematical model of ar-
tificial pneumoperitoneum, and lastly determine the location
and lesion parametrization model. This stage is the basis of
the entire algorithm, and also the most time-consuming stage. Figure 7. The mechanism diagram of the telecentric fixed-point
positioning mechanism. In the second stage, all allowable surgical incisions are ob-
tained from the first stage, and then the candidate incisions
are determined according to the two principles. The candi-
date base positions are obtained by the candidate incisions. According to the actual situation of the operating room, se-
lect one of the positions as the base position. Combine can-
didate incisions and the base position to determine the opti-
mum incision. In the third stage, the candidate entry angles are deter-
mined by combining the optimum incision, lesion location,
and initial entry angle. Determine the optimum angle accord-
ing to the observation direction principle. Since there may be
no direction in which the visual axis is perpendicular to the
plane τ, the minimum β is chosen as the optimum angle. Finally, the laparoscope arm preoperative planning algo-
rithm is completed, including the optimum incision and the
optimum angle. Figure 7. The mechanism diagram of the telecentric fixed-point
positioning mechanism. Figure 7. 3.3
The preoperative planning algorithm framework The mechanism diagram of the telecentric fixed-point
positioning mechanism. www.mech-sci.net/10/119/2019/ 3.2
The lesion parametrization model Yu et al.: Laparoscope arm automatic positioning 123 Figure 6. Flow chart of the laparoscope arm preoperative planning
l
i h Figure 6. Flow chart of the laparoscope arm preoperative planning
algorithm. Figure 5. The definition of lesion parameters. Figure 5. The definition of lesion parameters. 3.3
The preoperative planning algorithm framework 3.2
The lesion parametrization model 3.2
The lesion parametrization model The surgeon should be clear about the information of the
surgical site, including lesion location, lesion anatomy, and
surrounding tissues. At present, the conventional method is
imaging (radiology) test, and the lesion model and its sur-
rounding environment are obtained by the 3-D reconstruction
technology. Describe the relationship between lesion and in-
cision in parametric form, as shown in Fig. 5. Plane τ repre-
sents the target operation plane, a represents the normal vec-
tor of the plane τ, d represents the distance from the lesion
to the laparoscope, β represents the angle between laparo-
scope visual axis and a, γ represents the laparoscope devia-
tion angle. So, the two principles of laparoscope arm preop-
erative planning can be expressed as follows: (1) Observation
distance principle: laparoscope-to-target distance d = 75–
150 mm, d ≤the maximum joint variable of d7 (definition
in Fig. 3), and no barrier (Hanna et al., 1997b). (2) Observa-
tion direction principle: axis-to-target view angle, the smaller
β is, the better operative field is. When β = 0, the operative
field is optimum; in other words, the laparoscope visual axis
is perpendicular to the plane τ (Hanna and Cuschieri, 1999). Figure 3. The structure of the robotic arm. 2008; Oda et al., 2012), so the abdominal wall is simplified to
ellipsoid, defined as Eq. (1). The artificial pneumoperitoneal
coordinate frame is established by combining the patient’s
CT images and anatomy. According to anatomy, there are
three principal planes, namely the sagittal plane, the coronal
plane, and the transverse plane. In the coordinate frame, there
are also three reference planes, namely A plane, B plane,
and C plane. A plane coincides with the sagittal plane; B
plane coincides with the coronal plane; C plane is parallel to
the transverse plane, and the pneumoperitoneum is divided
equally by C plane. The origin of the coordinate frame is at
the intersection of three reference planes. xp-axis is defined
along the mediolateral direction; yp-axis is defined along the
superior-inferior direction; zp-axis is defined along the an-
teroposterior direction. In the coordinate frame, the mathe-
matical model of artificial pneumoperitoneum is established, Mech. Sci., 10, 119–131, 2019 www.mech-sci.net/10/119/2019/ L. Yu et al.: Laparoscope arm automatic positioning
Figure 5. The definition of lesion parameters. L. Yu et al.: Laparoscope arm automatic positioning
Figure 5. The definition of lesion parameters. L. 2. The link o1o4 is the shortest link:
(x −xc)2
(ac + lmax)2 +
(y −yc)2
(bc + lmax)2 ⩽1
x > ap
(11) (11) 3.6
The optimum incision and the optimum angle 3.6
The optimum incision and the optimum angle Pb (xb, yb, zb) is chosen as the base position, so the optimum
incisions are within allowable incisions circle with the Pb
as the origin and lmax as the radius. The optimum incisions
(red) are located on the intersection of candidate incisions
(green) and allowable incisions circle (black), as shown in
Fig. 11 and Eq. (12). The optimum angle of laparoscope en-
try is β = 0, that is, laparoscope visual line coincides with the
line relating incision to the lesion. To sum up, combined with
Sects. 3.5 and 3.6, the laparoscope arm preoperative planning
algorithm is completed. 2. The link o1o4 is the shortest link: 2. The link o1o4 is the shortest link:
0 < l ⩽a4
l + a2 ⩽a3 + a4 ⇒0 < l ⩽min(a4,a3 + a4 −a2 )
(9) (9) Figure 8. The schematic diagram of candidate incisions. 3. The link o1o4 is neither the longest nor the shortest link: First, candidate incisions are determined based on the
distance principle. Assume that the positions of the candi-
date incision and the lesion are Pi and Pl, respectively. If
d(PiPl) = |Pi −Pl| ≤d7max, Pi satisfies the distance princi-
ple. Second, based on the direction principle, the candidate
incisions are located on the generatrix of a right circular cone
with specific apex at Pl and aperture π −2α. So, candidate
incisions are incisions that satisfy the two principles. The fol-
lowing is a mathematical derivation of candidate incisions.
a4 < l < a2
a2 + a4 ⩽l + a3 ⇒max(a4,a2 + a4 −a3) < l < a2
(10) (10) According to Sect. 3.4, the allowable base range is an
ellipse. Compared with the projection of the candidate in-
cisions on the plane xpopyp, the center coordinate is un-
changed and the semi-major axis and semi-minor axis in-
crease lmax. The intersection of allowable base range and
non-interference area in the operating room is the candidate
base positions, as shown in Eq. (11). g
The Pl (xl, yl, zl) is obtained by imaging test; the candi-
date incisions are located on the intersection (red, Eq. 3) of
the abdominal wall (navy blue) and right circular cone with
specific apex at Pl (light blue), as shown in Fig. 8. The inter-
secting line is not a plane curve; it is projected to the plane
xpopyp for the convenience of research. The projection curve
(Eq. 4) is an ellipse whose expression can be obtained by fit-
ting four points on it. Go through Pl and make two planes
parallel to ypopzp and xpopzp, point m1, m2, m3 and m4 are
obtained, go through P z5
l
(xl, yl, z5) and make one plane par-
allel to xpopyp, point m5 and m6 are obtained, as shown in
Fig. 9 and Eqs. (5)–(7). The equation’s coefficients can be
obtained from any four points in the above six points, and
the remaining two points are used to verify the correctness
of them. 3.5
The candidate base positions Besides, the base position also affects the surgical incisions. Removing the prismatic joint, the telecentric fixed-point po-
sitioning mechanism is a 3-RRR planar redundant mecha-
nism. When o4 remains unchanged, it is simplified as a pla-
nar four-bar mechanism, as shown in Fig. 10. In this case, the
link length relationship determines whether the laparoscope
trajectory is a whole cone, which makes it possible to provide
the optimum operative field. In other words, o3o4 should be
rotated around o4 while o4 is unchanged, that is, the o3o4 is
a crank. Based on the conditions of crank existence, the link
length relationship is determined. Assume that the length of
link o1o2, o2o3, o3o4 and o4o1 are a2, a3, a4 and l, l de-
termines if there is a crank, which is discussed under three
cases, as shown in Eqs. (8)–(10). In summary, the distance
from base to fixed-point should be less than a2 + a3 −a4 to
ensure that laparoscope has a complete operative field.
(x −xc)2
a2c
+ (y −yc)2
b2c
= 1
(x −xb)2 + (y −yb)2 < l2
max
(12) (12) Given a set of parameters based on the actual situation, the
steps of the algorithm are described in detail, a2 = 220 mm,
a3 = 220 mm, a4 = 150 mm, α = 45◦, Pl = (0.35,0.2,0.3),
P z5
l
= (0.35,0.2,1.5) (in the op −xpypzp coordinate frame). L. Yu et al.: Laparoscope arm automatic positioning 124 Figure 8. The schematic diagram of candidate incisions. 1. The link o1o4 is the longest link: 1. The link o1o4 is the longest link: 1. The link o1o4 is the longest link: l ⩾a2
l + a4 ⩽a2 + a3 ⇒a2 ⩽l ⩽a2 + a3 −a4
(8) (8) 2. The link o1o4 is the shortest link: 3.4
The candidate incisions The telecentric fixed-point positioning mechanism has four
degrees of freedom; the mechanism diagram is shown in
Fig. 7. o4 is the telecentric fixed-point, o5 is the end of a
laparoscope, and α is determined by the remote center of mo-
tion mechanism. The prismatic joint is used to adjust the ver-
tical position of o4, and the three revolute joints are used to adjust the horizontal position. Removing the prismatic joint,
it is a planar redundant mechanism. When o4 remains un-
changed, the motion trajectory of the laparoscope is a right
circular cone with specific apex at o4 and aperture π −2α. adjust the horizontal position. Removing the prismatic joint,
it is a planar redundant mechanism. When o4 remains un-
changed, the motion trajectory of the laparoscope is a right
circular cone with specific apex at o4 and aperture π −2α. www.mech-sci.net/10/119/2019/ Mech. Sci., 10, 119–131, 2019 www.mech-sci.net/10/119/2019/ 3.6.1
Step 1 Determine candidate incisions Take the data in Sect. 3.1, 0 ≤d7 ≤320 mm, d(PiPl)max =
sqrt
(h2 + (ap
(1 + sqrt(c2
p −h2)/cp))2) = 310.7 mm Take the data in Sect. 3.1, 0 ≤d7 ≤320 mm, d(PiPl)max =
sqrt
(h2 + (ap
(1 + sqrt(c2
p −h2)/cp))2) = 310.7 mm Mech. Sci., 10, 119–131, 2019 www.mech-sci.net/10/119/2019/ 125 L. Yu et al.: Laparoscope arm automatic positioning Figure 9 The projection of three planes Figure 9. The projection of three planes. Figure 9. The projection of three planes. Figure 10. The schematic diagram of the simplified mechanism. on the plane xpopyp is shown in Eq. (14). on the plane xpopyp is shown in Eq. (14).
x2
1.552 +
y2
1.452 +
z2
2.272 = 1
(z −0.3)2 =
1
tan245
◦
h
(x −0.35)2 + (y −0.2)2i
(13)
(x −0.13)2
1.34
+ (y −0.07)2
1.23
= 1
(14) (13) (14) 3.6.2
Step 2 Determine base position 3.6.2
Step 2 Determine base position Figure 10. The schematic diagram of the simplified mechanism. The candidate base positions are located on the curve, as
shown in Eq. (15). Within the allowable base range, choose a
base position Pb = (2.3,−0.2, zb), zb is determined accord-
ing to the condition of the operating room. (x −0.13)2
(1.16 + 2.9)2 + (y −0.07)2
(1.11 + 2.9)2 ⩽1
x > 1.55
(15) (sqrt = square root), the result shows that any point on
the abdominal wall can be used as a candidate incision. The
candidate incisions are located on the curve, as shown in
Eq. (13). According to Sect. 3.4, the projection of the curve (15) Mech. Sci., 10, 119–131, 2019 Mech. Sci., 10, 119–131, 2019 Mech. Sci., 10, 119–131, 2019 www.mech-sci.net/10/119/2019/ L. Yu et al.: Laparoscope arm automatic positioning L. Yu et al.: Laparoscope arm automatic positioning 126 L. Yu et al.: Laparoscope arm automatic positioning
Figure 13. The agent–environment interaction in reinforcement
learning. Figure 11. The schematic diagram of optimum incisions. Figure 11. The schematic diagram of optimum incisions. Figure 13. The agent–environment interaction in reinforcement
learning. 4.1
Problem description Figure 11. The schematic diagram of optimum incisions. Reinforcement learning describes the set of learning prob-
lems where an agent should learn how to map states to ac-
tions in an environment to maximize the defined reward func-
tion. Throughout the learning process, an agent is not told
which actions to take but instead should find out which ac-
tion yield the most reward by trying various actions. In most
cases, actions may affect not only the immediate reward but
also the next state, and through that all subsequent rewards. In solving practical problems, it should define a reasonable
reward function to compute the reward for taking actions and
have a goal relating to the state of the environment. Also, it
should quantify all the variables the environment describes
and have access to these variables at each step or state. Figure 12. The sketch map of the optimum incision and angle. In this paper, the agent is the 3-RRR planar redundant
mechanism which is a simplified model of telecentric fixed-
point positioning mechanism plus laparoscope. The environ-
ment is the lesion and the surgical incision obtained through
the preoperative planning algorithm. The actions are the
movement of three revolute joints. The agent–environment
interaction is shown in Fig. 13. Figure 12. The sketch map of the optimum incision and angle. www.mech-sci.net/10/119/2019/ 4.3
Reward function construction In the training process, telecentric fixed-point (marked point)
position and lesion location are taken as the input of the
DDPG algorithm. The fixed-point is obtained by a depth
camera, the optimum incision, the optimum angle and the
base position are obtained by the preoperative planning algo-
rithm. The DDPG algorithm that combines the algorithm can
learn policies directly from the inputs, to achieve laparoscope
arm automatic positioning for the robot-assisted laparoscopic
surgery. The reward function is essential for the algorithm
to learn policies successfully. It consists of intermediate re-
ward and final reward, where the former is given a continu-
ous, guided negative reward when the task is not completed,
and the latter is given a positive reward that is one to two
orders of magnitude larger than the former when the task is
completed. The continuous reward function can make con-
vergence of the algorithm better. TensorFlow is used in the code for high-performance nu-
merical computation. The simulations use Adam (Kingma
and Ba, 2015) for learning neural network parameters with a
learning rate of 10−5 for the actor and critic. For Q it includes
L1 weight decay of 0.1, L2 weight decay of 10−3 and a dis-
count factor of γ = 0.9. For the soft target updates, it uses
τ = 0.01. The neural networks use the rectified non-linearity
for all hidden layers (Glorot et al., 2011). The networks have
three hidden layers with 900, 900 and 60 units respectively,
and the final output layer of the actor is a tanh layer, to bound
the actions. The actions are not included until the 3rd hidden
layer of Q. The layers weights and biases of both the actor
and critic are initialized from a uniform distribution [−x, x],
where x = sqrt (6./(in + out)). It trains with minibatch sizes
of 16, and it uses a replay buffer size of 6 × 104. The behav-
ior policy during training is ε-greedy with ε annealed linearly In the op −xpypzp coordinate frame, the fixed-point po-
sition is Pf (xf, yf, zf), the incision position is Pi (xi, yi,
zi), the laparoscope end position is Pe (xe, ye, ze), and the
lesion location is Pl (xl, yl, zl). The goal of the task is
|PfPi| + |PePl| =0 (lsinα (definition in Fig. 7) is equal to
|PiPl| for programming convenience.). 5.1
Simulation details (20) The environment is simulated using Pyglet, including a le-
sion point, a surgical incision and a simplified model of the
telecentric fixed-point positioning mechanism. For this envi-
ronment, a lesion point is randomly specified within a rea-
sonable range, an incision and a base location are obtained
by the preoperative planning algorithm. Batch normalization
is used on the state input, all layers of the actor network and
all layers of the critic network before the action input. In this
way, it can learn effectively across tasks with different types
of units, without needing to ensure the units are within a set
range manually. 5
Simulation and results θQ′ ←τθQ + (1 −τ)θQ′
θµ′ ←τθµ + (1 −τ)θµ′
(20) 4.4
States description celerate deep network training and improve the accuracy of
the model (Ioffe and Szegedy, 2015). 4.4
States description To improve the convergence of the algorithm, the state vari-
ables also play a crucial role in addition to the reward func-
tion. If state variables can adequately present the environ-
ment, the algorithm can learn policies quickly. Because the
image from the depth camera contains all the state informa-
tion of the environment, it is reasonable to use the image di-
rectly as input. However, due to the limitations of the hard-
ware, the processing image data is very slow. To speed up
training of the algorithm, it uses a low-dimensional states
description, such as joint variables and positions, instead of
high-dimensional renderings of the environment. DDPG contains a parameterized actor function µ(s|θµ)
and critic network Q(s, a|θQ) with weights θµ and θQ. The critic network is learned using the Bellman equation
(Eqs. 17–18) to make the L(θQ) smaller and smaller. In other
words, Q(s, a|θQ) gets closer to the actual value. L(θQ) = Est∼ρβ,at∼β,rt∼E
Q
st,at|θQ
−yt
2
(17) (17) where The algorithm is to make the laparoscope arm move to the
target position, so the joint variables are used as the state
variables. However, from the training results, these variables
cannot adequately describe the environment; in other words,
the algorithm cannot achieve the laparoscope arm automatic
movement. So, the distance from telecentric fixed-point to
incision, the distance from laparoscope end to the lesion, and
whether the target is reached are added to the state variables. The experimental results of these two state variables are de-
scribed in Sect. 5.2. yt = r (st,at) + γ Q
st+1,µ(st+1)|θQ
(18) (18) The actor function is updated by the chain rule (Eq. 19) to
the expected return from the start distribution J with respect
to the actor parameters. ∇θµJ ≈Est∼ρβ
∇θµQ(s,a|θQ)|s=st,a=µ(st|θµ)
= Est∼ρβ
∇aQ(s,a|θQ)|s=st,a=µ(st)∇θµµ(s|θµ)|s=st
(19) (19) Every n steps DDPG updates the target networks of actor
and critic using “soft” target updates (Eq. 20), rather than
directly copying the weights. 4.2
Deep deterministic policy gradient (DDPG) 3.6.3
Step 3 Determine the optimum incision and the
optimum angle In this paper, laparoscope arm automatic positioning is
achieved by DDPG, which is a model-free, off-policy actor-
critic algorithm based on the deterministic policy gradient
(DPG) (Silver et al., 2014). Deep neural network (DNN)
function approximators were used to estimate the action-
value function. Thus, the algorithm can learn policies in high-
dimensional, continuous action spaces. First, determine xi based on the surgical needs, body con-
dition and surgeon’s operating habits, calculate yi based on
Eq. (16), calculate zi based on the mathematical model of
pneumoperitoneum, the optimum incision is (xi, yi, zi). Sec-
ond, the optimum visual axis direction is the line connecting
the optimum incision to the lesion. The optimum incision and
optimum angle are shown in Fig. 12. Based on DPG, DDPG combines the ideas underlying
the success of Deep Q Network (DQN) (Mnih et al., 2013,
2015). It can learn value functions stably and robustly due
to two aspects. First, the network is trained off-policy with
samples from a replay buffer to minimize correlations be-
tween samples. Second, the network is trained with a target
Q network to give consistent targets during temporal differ-
ence backups. Meanwhile, batch normalization is used to ac-
(x −0.13)2
1.34
+ (y −0.07)2
1.23
= 1
(x −2.3)2 + (y + 0.2)2 < 2.92
(16) (16) Mech. Sci., 10, 119–131, 2019 www.mech-sci.net/10/119/2019/ 127 L. Yu et al.: Laparoscope arm automatic positioning www.mech-sci.net/10/119/2019/ 4.3
Reward function construction The intermediate re-
ward is −(|PfPi|+|PePl|) and is normalized to [−1,0] inter-
val. The final reward is 10. www.mech-sci.net/10/119/2019/ Mech. Sci., 10, 119–131, 2019 L. Yu et al.: Laparoscope arm automatic positioning
Figure 15. The steps to target with states descriptor one. Figure 16. The total reward per episode with states descriptor two. 128
Figure 14. The total reward per episode with states descriptor one. L. Yu et al.: Laparoscope arm automatic positioning
Figure 15. The steps to target with states descriptor one. 128
Figure 14. The total reward per episode with states descriptor one. 128 Figure 14. The total reward per episode with states descriptor one. Figure 15. The steps to target with states descriptor one. Figure 15. The steps to target with states descriptor one. Figure 16. The total reward per episode with states descriptor two. Figure 16. The total reward per episode with states descriptor two. from 1 to 0.1 over the first hundred episodes and fixed at 0.1
after that. The simulations train for a total of 2000 episodes;
every episode is terminated if the goal is not completed after
600 steps. 5.2
Simulation results Two simulations are set up to evaluate the performance of
the improved method applied to laparoscope arm automatic
positioning for the robot-assisted laparoscopic surgery. The
two simulations make one change to states description during
training only, and use the same network architecture, learn-
ing algorithm and hyperparameters settings. States descrip-
tor one is three joint variables and states descriptor two is
the former plus the distance from fixed-point to incision, the
distance from laparoscope end to the lesion, and whether the
target is reached. Figure 16. The total reward per episode with states descriptor two. The two simulations evaluate the policy periodically dur-
ing training by testing it without exploration noise. The im-
proved method with 3 action dimensions and 20 state dimen-
sions runs ten times in the simulated environment. Perfor-
mance after training across the environment for at most 2000
episodes. The results of ten training sessions report both total
reward per episode and steps to target, as shown in Figs. 14–
17. The solid line in the figure represents the average over ten
sessions, the upper boundary of the shadow part represents
the maximum over ten sessions, and the lower boundary rep-
resents the minimum value. results illustrate the states descriptor two is outperformed
states descriptor one, the latter does not enable the agent to
converge to a good solution, but the former can do it. In other
words, the improved method which uses the states descriptor
two can learn the right policies on laparoscope arm automatic
positioning. Mech. Sci., 10, 119–131, 2019 6
Discussion The steps to target with states descriptor two. The proposed algorithm is designed to simulate the actual
clinical procedure of robot-assisted surgery or applied to a
virtual surgery training system, and a standardized procedure
is proposed for preoperative planning. By taking LC as an
example, the results indicate that the port placement and la-
paroscope entry angle selection have satisfying performance,
especially for less experienced surgeons. The automatic positioning algorithm provides a theoretical
basis for the laparoscope arm preoperative planning of robot-
assisted laparoscopic surgery. It avoids the disadvantage of
the heuristic method based on surgeon experience, and it also
simplifies the preoperative planning process and reduces the
operation time. However, the algorithm is implemented in a
virtual environment, and there is a certain gap with the actual
system. Therefore, how to implement the algorithm in the
actual system is the primary direction of subsequent research. p
y
p
g
Preoperative laparoscope arm automatic positioning is
achieved based on the DDPG. In this algorithm, the states
descriptor plays a crucial role and affects the performance of
the algorithm. From the results, the states descriptor two is
outperformed states descriptor one. Although the controller
does not learn a reasonable strategy directly from states de-
scriptor one, with the evolution of episodes, the controller
still improves compared to the initial. Therefore, it is crucial
to select states descriptor reasonably. The controller learns a
reasonable strategy from states descriptor two, but there is
room to reduce the steps of the target, to improve the learn-
ing efficiency of the controller. Furthermore, the laparoscope
arm automatic positioning is independent of robot configura-
tion and can be extended to any surgical robot system. Data availability. The data in this study can be requested from the
corresponding author. Author contributions. LY, XY, XC and FZ discussed and decided
on the methodology in the study. The preoperative planning algo-
rithm, the reinforcement learning algorithm and simulations have
been performed by XY, XC and FZ. LY completed literature review
and overall plan. This method successfully learns a controller in simulation,
and the next step is to study to learn a controller in real robots
without a lot of time training, and the method can be ex-
tended to the preoperative planning of other operations or
even other surgical procedures. 6
Discussion Thus, the implementation of
the algorithm for robot-assisted surgery can further realize
telesurgery, thereby improving the medical level in many ar-
eas. Competing interests. The authors declare that they have no con-
flict of interest. Acknowledgements. The paper is supported by the Natural Sci-
ence Foundation of Heilongjiang Province (Grand No. F2015034). We also greatly appreciate the efforts of the reviewers and our col-
leagues. 6
Discussion The preoperative planning algorithm, based on the artificial
pneumoperitoneum model and the lesion parametrization
model, appears to offer significant improvements in plan-
ning time and quality for robot-assisted laparoscopic surgery
over experience-based method or literature-based method. The distance principle and the direction principle ensure that
the proposed algorithm can meet the surgeon’s surgical re-
quirements. Furthermore, preoperative planning does not re-
quire an additional landmark on the abdominal wall or par-
ticular patient positioning. Figure 14 shows that the average of total reward per
episode is stabilized to negative and only a few episodes total
reward are positive. Figure 15 shows that the steps to target
are always 600. These two figures show that it never reaches
the goal. Figure 16 shows that the average of total reward
per episode increases from −300 to about 120. After 400
episodes, the total reward converges to around 120. Figure 17
shows the steps to target stabilizes at about 150. These two
figures show that it reaches the goal after 400 episodes. The Mech. Sci., 10, 119–131, 2019 www.mech-sci.net/10/119/2019/ 129 L. Yu et al.: Laparoscope arm automatic positioning L. Yu et al.: Laparoscope arm automatic positioning L. Yu et al.: Laparoscope arm automatic positioning
Figure 17. The steps to target with states descriptor two. principle and the direction principle. According to the two
principles, the laparoscope arm preoperative planning algo-
rithm is divided into two parts, the optimum incision and
the optimum angle of laparoscope entry. A set of parame-
ters based on the actual situation is given to verify the ef-
fectiveness of the algorithm. Preoperative laparoscope arm
automatic positioning is achieved by the improved method
which combines the preoperative planning algorithm with
the DDPG algorithm. The improved method takes the fixed-
point position captured by a depth camera and the lesion lo-
cation obtained by imaging test as input. Based on the input
information, optimum incision and optimum angle are ob-
tained through the algorithm, and then the laparoscope arm
can automatically move to the target position. Compared to
the traditional method, kinematics is not used to calculate
the motor movements, so that it can reduce errors caused
by inaccuracy of kinematic parameters and improve the ef-
fectiveness of preoperative planning. The simulation results
show that the improved method can realize preoperative la-
paroscope arm automatic positioning and it is also robust. Figure 17. www.mech-sci.net/10/119/2019/ Competing interests. The authors declare that they have no con-
flict of interest. 7
Conclusions This paper completes the preoperative planning by ana-
lyzing the surgical procedures and surgical environment
of robot-assisted laparoscopic surgery. Based on the lesion
parametrization model, two principles of laparoscope arm
preoperative planning are designed, including the distance Review statement. This paper was edited by Jinguo Liu and re-
viewed by Yi Yang and two anonymous referees. Mech. Sci., 10, 119–131, 2019 www.mech-sci.net/10/119/2019/ L. Yu et al.: Laparoscope arm automatic positioning 130 References Hayashibe,
M.,
Suzuki,
N.,
Hashizume,
M.,
Konishi,
K.,
and
Hattori,
A.:
Robotic
surgery
setup
simulation
with
the integration of inverse-kinematics computation and med-
ical imaging, Comput. Methods Progr. Biomed., 83, 63–72,
https://doi.org/10.1016/j.cmpb.2006.04.010, 2006. Austad, A., Elle, O. J., and Røtnes, J. S.: Computer-aided
planning of trocar placement and robot settings in robot-
assisted
surgery,
Int. Congr. Series,
1230,
1020–1026,
https://doi.org/10.1016/S0531-5131(01)00179-0, 2001. Ioffe, S. and Szegedy, C.: Batch normalization: accelerating deep
network training by reducing internal covariate shift, arXiv
preprint, arXiv:1502.03167, 2015. Azimian, H., Breetzke, J., Trejos, A. L., Patel, R. V., Naish, M. D., Peters, T., Moore, J., Wedlake, C., and Kiaii, B.: Preoper-
ative planning of robotics-assisted minimally invasive coronary
artery bypass grafting, in: 2010 IEEE International Conference
on Robotics and Automation, Anchorage, United States, 3–7
May 2010, 1548–1553, 2010. James, S. and Johns, E.: 3D simulation for robot arm control with
deep Q-learning, arXiv preprint, arXiv:1609.03759, 2016. Kingma, D. P. and Ba, L. J.: Adam: A method for stochastic op-
timization. in: International Conference on Learning Represen-
tations 2015, San Diego, United States, 7–9 May 2015, 1–15,
2015. Badani, K. K., Muhletaler, F., Fumo, M., Kaul, S., Peabody, J. O.,
Bhandari, M., and Menon, M.: Optimizing robotic renal surgery:
the lateral camera port placement technique and current results, J. Endourol., 22, 507–510, https://doi.org/10.1089/end.2007.0228,
2008. Konietschke, R., Bodenmüller, T., Rink, C., Schwier, A., Bäuml, B.,
and Hirzinger, G.: Optimal setup of the DLR MiroSurge teler-
obotic system for minimally invasive surgery, in: 2011 IEEE In-
ternational Conference on Robotics and Automation, Shanghai,
China, 9–13 May 2011, 3435–3436, 2011. Bauernschmitt, R., Feuerstein, M., Traub, J., Schirmbeck, E. U., Klinker, G., and Lange, R.: Optimal port placement and
enhanced guidance in robotically assisted cardiac surgery,
Surg. Endosc., 21, 684–687, https://doi.org/10.1007/s00464-
006-9057-z, 2007. Ma, R. Q., Wang, W. D., Dong, W., and Du, Z. J.: Preoperative
positioning analysis of the celiac minimally invasive surgery
robotic system based on an improved gradient projection algo-
rithm, Robot, 32, 156–163, 2014. 7
Conclusions Cestari, A., Buffi, N. M., Scapaticci, E., Lughezzani, G., Salonia,
A., Briganti, A., Rigatti, P., Montorsi, F., and Guazzoni, G.: Sim-
plifying patient positioning and port placement during robotic-
assisted laparoscopic prostatectomy, Eur. Urol., 57, 530–533,
https://doi.org/10.1016/j.eururo.2009.11.028, 2010. Mnih, V., Kavukcuoglu, K., Silver, D., Graves, A., Antonoglou, I.,
Wierstra, D., and Riedmiller, M.: Playing atari with deep rein-
forcement learning, arXiv preprint, arXiv:1312.5602, 2013. Mnih, V., Kavukcuoglu, K., Silver, D., Rusu, A. A., Veness, J.,
Bellemare, M. G., Graves, A., Riedmiller, M., Fidjeland, A. K.,
Ostrovski, G., Petersen, S., Beattie, C., Sadik, A., Antonoglou, I.,
King, H., Kumaran, D., Wierstra, D., Legg, S., and Hassabis, D.:
Human-level control through deep reinforcement learning, Na-
ture, 518, 529–533, https://doi.org/10.1038/nature14236, 2015. Chen,
X. R.:
How
to
establish
pneumoperitoneum
safely
in
laparoscopic
surgery,
J. Abdomin. Surg.,
12,
12–13,
https://doi.org/10.3969/j.issn.1003-5591.1999.01.006, 1999. Ferzli, G. S. and Fingerhut, A.: Trocar placement for la-
paroscopic
abdominal
procedures:
a
simple
standard-
ized
method,
J. Am. Coll. Surgeons,
198,
163–173,
https://doi.org/10.1016/j.jamcollsurg.2003.08.010, 2004. Mohammadi, B., Kerzel, M., Görner, M., Zamani, M. A., Eppe,
M., and Wermter. S.: Neural end-to-end learning of reach for
grasp ability with a 6-dof robot arm, in: Workshop on Machine
Learning in Robot Motion Planning–2018 IEEE/RSJ Interna-
tional Conference on Intelligent Robots and Systems, Madrid,
Spain, 1–5 October 2018, 1–3, 2018. Glorot, X., Bordes, A., and Bengio, Y.: Domain adaptation for
large-scale sentiment classification: A deep learning approach,
in: ICML’11 Proceedings of the 28th International Conference on
International Conference on Machine Learning, Bellevue, United
States, 28 June–2 July 2011, 513–520, 2011. Mulier, J., Coenegrachts, K., and Moortele, K. V. D.: CT analysis of
the elastic deformation and elongation of the abdominal wall dur-
ing colon inflation for virtual coloscopy, Eur. J. Anaesthesiol., 25,
42, https://doi.org/10.1097/00003643-200805001-00132, 2008. Gu, S., Holly, E., Lillicrap, T., and Levine, S.: Deep reinforcement
learning for robotic manipulation with asynchronous off-policy
updates, in: 2017 IEEE International Conference on Robotics
and Automation, Singapore, Singapore, 29 May–3 June 2017,
3389–3396, 2017. Oda, M., Qu, J. D., Nimura, Y., Kitasaka, T., Misawa, K., and
Mori, K.: Evaluation of deformation accuracy of a virtual pneu-
moperitoneum method based on clinical trials for patient-specific
laparoscopic surgery simulator, Medical Imaging 2012: Image-
Guided Procedures, Robotic Interventions, and Modeling, 8316,
8316G, https://doi.org/10.1117/12.911701, 2012. Hanna, G. and Cuschieri, A.: Influence of the optical axis-to-target
view angle on endoscopic task performance, Surg. Endosc., 13,
371–375, https://doi.org/10.1007/s004649900992, 1999. 7
Conclusions Hanna, G., Shimi, S., and Cuschieri, A.: Optimal port locations for
endoscopic intracorporeal knotting, Surg. Endosc., 11, 397–401,
https://doi.org/10.1007/s004649900374, 1997a. Otte, S., Zwiener, A., Hanten, R., and Zell, A.: Inverse recur-
rent models–an application scenario for many-joint robot arm
control, in: Artificial Neural Networks and Machine Learning–
International Conference on Artificial Neural Networks 2016,
Barcelona, Spain, 6–9 September 2016, 149–157, 2016. Hanna, G., Shimi, S., and Cuschieri, A.: Influence of direc-
tion of view, target-to-endoscope distance and manipulation an-
gle on endoscopic knot tying, Brit. J. Surg., 84, 1460–1464,
https://doi.org/10.1111/j.1365-2168.1997.02835.x, 1997b. Phaniteja, S., Dewangan, P., Guhan, P., Sarkar, A., and Krishna, K. M.: A deep reinforcement learning approach for dynamically sta-
ble inverse kinematics of humanoid robots, in: 2017 IEEE In-
ternational Conference on Robotics and Biomimetics, Macau,
China, 5–8 December 2017, 1818–1823, 2017. Hayashibe, M., Suzuki, N., Hashizume, M., Kakeji, Y., Kon-
ishi, K., Suzuki, S., and Hattori, A.: Preoperative planning
system for surgical robotics setup with kinematics and hap-
tics, The Int. J. Med. Robot. Comput. Assist. Surg., 1, 76–85,
https://doi.org/10.1002/rcs.18, 2005. Mech. Sci., 10, 119–131, 2019 www.mech-sci.net/10/119/2019/ L. Yu et al.: Laparoscope arm automatic positioning 131 Sun, L. W., Meer, F. V., Schmid, J., Bailly, Y., Thakre, A. A., and Yeung, C. K.: Advanced da Vinci surgical sys-
tem simulator for surgeon training and operation planning,
The Int. J. Med. Robot. Comput. Assist. Surg., 3, 245–251,
https://doi.org/10.1002/rcs.139, 2007. Pick, D. L., Lee, D. I., Skarecky, D. W., and Ahlering,
T. E.:
Anatomic
guide
for
port
placement
for
daVinci
robotic radical prostatectomy, J. Endourol., 18, 572–575,
https://doi.org/10.1089/end.2004.18.572, 2004. Silver, D., Lever, G., Heess, N., Degris, T., Wierstra, D., and
Riedmiller, M.: Deterministic policy gradient algorithms, in:
ICML’14 Proceedings of the 31st International Conference on
International Conference on Machine Learning, Beijing, China,
21–26 June 2014, 387–395, 2014. Wang,
W.,
Wang,
W. D.,
Dong,
W.,
Du,
Z. J.,
and
Sun,
Y. P.:
A
preoperative
planning
algorithm
based
on
dexterity
and
collaborationspace
for
the
robot-
assisted minimally invasive surgery, Robot, 38, 208–216,
https://doi.org/10.13973/j.cnki.robot.2016.0208, 2016. Sun, L. W. and Yeung, C. K.: Port placement and pose selection
of the da Vinci surgical system for collision-free intervention
based on performance optimization, in: 2007 IEEE/RSJ Interna-
tional Conference on Intelligent Robots and Systems, San Diego,
United States, 29 October–2 November 2007, 1951–1956, 2007. Yu, L. T., Wang, Z. Y., Sun, L. Q., Wang, W. www.mech-sci.net/10/119/2019/ L. Yu et al.: Laparoscope arm automatic positioning 7
Conclusions J., and Wang, L.: Re-
search on preoperative positioning analysis of instrument arms
for minimally invasive surgical robot, in: 2014 IEEE Interna-
tional Conference on Mechatronics and Automation, Tianjin,
China, 3–6 August 2014, 1269–1275, 2014. Mech. Sci., 10, 119–131, 2019 www.mech-sci.net/10/119/2019/
|
https://openalex.org/W2803252074
|
https://europepmc.org/articles/pmc5871888?pdf=render
|
English
| null |
Identification and DNA annotation of a plasmid isolated from Chromobacterium violaceum
|
Scientific reports
| 2,018
|
cc-by
| 6,572
|
Identification and DNA annotation
of a plasmid isolated from
Chromobacterium violaceum
Daniel C. Lima1,2, Lena K. Nyberg3, Fredrik Westerlund3 & Silvia R. Batistuzzo de Medeiros2 Received: 14 December 2017
Accepted: 12 March 2018
Published: xx xx xxxx Daniel C. Lima1,2, Lena K. Nyberg3, Fredrik Westerlund3 & Silvia R. Batistuzzo de Medeiros2 Chromobacterium violaceum is a ß-proteobacterium found widely worldwide with important
biotechnological properties and is associated to lethal sepsis in immune-depressed individuals. In this
work, we report the discover, complete sequence and annotation of a plasmid detected in C. violaceum
that has been unnoticed until now. We used DNA single-molecule analysis to confirm that the episome
found was a circular molecule and then proceeded with NGS sequencing. After DNA annotation, we
found that this extra-chromosomal DNA is probably a defective bacteriophage of approximately 44
kilobases, with 39 ORFs comprising, mostly hypothetical proteins. We also found DNA sequences that
ensure proper plasmid replication and partitioning as well as a toxin addiction system. This report
sheds light on the biology of this important species, helping us to understand the mechanisms by
which C. violaceum endures to several harsh conditions. This discovery could also be a first step in the
development of a DNA manipulation tool in this bacterium. Chromobacterium violaceum is a Gram-negative facultative anaerobe bacillus belonging to the Neisseriaceae fam-
ily1. This free-living ß-proteobacterium reside mainly around tropical and sub-tropical regions. The study of C. violaceum started in the 1970s, focusing on its potential in pharmacology and industry for the production of
antibiotics, anti-tumoral substances, biopolymers and others organic compounds (reviewed in refs2–4). C. viola-
ceum is also an opportunistic pathogen that can cause severe infections and lead to sepsis and sometimes death
in immuno-depressed individuals5,6. p
In 2003, the complete genome of C. violaceum was sequenced and many genes related to stress adaptability
were identified. This led to a great number of studies of how the bacterium copes with environmental chal-
lenges7–11. Many studies focusing on understanding the mechanisms of quorum sensing in C. violaceum make this
organism an important model species12–14.fi Despite the great interest in C. violaceum and the sequencing of its entire genome7,8,15, efficient methods to
modify its genome are still not developed. For example, a study reported genetic transformation of C. violaceum16
but this methodology proved to be irreproducible by many groups. More recently, a group succeeded in generat-
ing mutants in C. violaceum using conjugation17. This method is laborious and mutants often revert. Therefore,
there is a demand to develop more efficient tools to conduct genetic studies in C. violaceum. Identification and DNA annotation
of a plasmid isolated from
Chromobacterium violaceum
Daniel C. Lima1,2, Lena K. Nyberg3, Fredrik Westerlund3 & Silvia R. Batistuzzo de Medeiros2 pfi
g
Here, we used single DNA molecule analysis and next-generation sequencing to identify a plasmid in C. viol-
aceum strain ATCC 12472. The presence of this 44,212 bp plasmid has been unnoticed until now and its charac-
terization may help building a shuttle vector that would greatly facilitate the development of genome engineering
tools for C. violaceum. www.nature.com/scientificreports www.nature.com/scientificreports www.nature.com/scientificreports Received: 14 December 2017
Accepted: 12 March 2018
Published: xx xx xxxx Experimental Procedures p
Plasmid isolation. Four isolated colonies of C. violaceum ATCC 12472 were inoculated in four flasks con-
taining 400 mL of LB medium each for 16–18 h. The cultures were centrifuged at 4 °C, 5 minutes, 7441 × g. The
pellets were resuspended with 25 mL of Ressuspension Buffer (50 mM Tris-HCl, 10 mM EDTA, RNAse 100 μg/
mL, pH 8.0), and then 25 mL of Lysis Buffer (SDS 1%; 0.2 M NaOH) was added, with 5 minutes of room tempera-
ture incubation. The plasmid DNA was precipitated by adding 25 mL of 3 M Potassium Acetate, pH 5.5, followed
by centrifugation at 22789 × g, 10 minutes at 4 °C. The supernatant was transferred to a new tube and 0.7 volume
of isopropanol was added. After one more step of centrifugation (22789 × g, 10 minutes, 4 °C), the pellets were 1Instituto Federal de Educação, Ciência e Tecnologia do Rio Grande do Norte, Natal, Brazil. 2Laboratório de Biologia
Molecular e Genômica, Universidade Federal do Rio Grande do Norte, Natal, Brazil. 3Department of Biology and
Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden. Correspondence and requests for
materials should be addressed to S.R.B.d.M. (email: sbatistu@gmail.com) SCIENTIFIC Reports | (2018) 8:5327 | DOI:10.1038/s41598-018-23708-5 1 www.nature.com/scientificreports/ Figure 1. Restriction digestion pattern from two independent preparations of the episome. The asterisk denotes
the band corresponding to the plasmid. The restriction enzyme used is mentioned on top of each lane. 0.8%
TBE Agarose gel stained with ethidium bromide. Figure 1. Restriction digestion pattern from two independent preparations of the episome. The asterisk denotes
the band corresponding to the plasmid. The restriction enzyme used is mentioned on top of each lane. 0.8%
TBE Agarose gel stained with ethidium bromide. re-suspended with 1 mL of TE Buffer and one volume of phenol:chloroform was added. After centrifugation
(22789 × g, 10 minutes, 4 °C), the aqueous phase was transferred to a new tube and the DNA was precipitated with
one volume of isopropanol. Finally, the DNA was washed with 80% Ethanol and the four independent prepara-
tions were re-suspended with 500 μL of TE. re-suspended with 1 mL of TE Buffer and one volume of phenol:chloroform was added. After centrifugation
(22789 × g, 10 minutes, 4 °C), the aqueous phase was transferred to a new tube and the DNA was precipitated with
one volume of isopropanol. Experimental Procedures Finally, the DNA was washed with 80% Ethanol and the four independent prepara-
tions were re-suspended with 500 μL of TE. In order to certify that our preparation was free of genomic DNA we isolated the band containing the plasmid
and digested the agarose using ß-agarase (NEB catalog # - M0392S) according to the manufacturer’s instructions. Optical analysis of DNA in nanochannels. The optical DNA mapping of the single plasmid molecules
were performed as described in ref.18. Using a combination of a DNA fluorescent dye (YOYO-1) and Netropsin,
an antibiotic that binds specifically to AT DNA regions, this technique allows the acquisition of DNA barcode
images, with dark and bright regions corresponding to AT-rich and GC-rich regions respectively19. In this
way, the pattern of the emission intensity reflects the sequence of the DNA molecule, with a resolution on the
kilobasepair length scale. The nanofluidic chips were fabricated in fused silica, using conventional techniques,
as described in ref.20. All the data was recorded, using a Zeiss AxioObserver. Z1 microscope equipped with a
100× TIRF oil immersion objective (NA = 1.46) from Zeiss and a Photometrics Evolve EMCCD camera. NGS Sequencing and Assembly. The DNA was quantified using Qubit Fluorometric Quantitation and
the quality was checked on an agarose gel. The library was prepared using TruSeq Nano DNA Sample Preparation
Kit (Illumina) according to the manufacturer’s instructions and then sequenced on the Illumina MiSeq at Fasteris
SA. For the base-calling, the CASAVA pipeline 1.8 was used. De novo genomic assembly was made using VELVET
v1.2.10 and Burrows-Wheeler Alignment Tool (v0.5.9) for mapping. Plasmid annotation and comparison. The annotation was made using Glimmer (v3.02b), a software
built to find genes in bacteria, archaea and viruses. Bacteria/archaea genetic code and circular topology were
chosen. The search for homology of the whole pChV1 sequence was made using the BLASTn program against
non-redundant (NR) NCBI database and against a specific bacteriophage database (unclassified bacterio-
phages – taxid: 12333), also from NCBI. Comparison of the predicted ORFs in genomic databases was made
using BLASTx. Hits with more than 50% coverage and with the highest BitScore were picked. Search for tRNAs
was made using the online version of tRNAscan-SE v1.21 in default mode. DNA inverted repeated sequences
were obtained using Einverted (http://emboss.bioinformatics.nl/cgi-bin/emboss/einverted). The search for pal-
indromic DNA was made using the MEME web-tool21. GC content profile and GC-skew were obtained using
GC-Profile22 and GenSkew (http://genskew.csb.univie.ac.at/), respectively. Experimental Procedures Data availability. The pChV1 complete sequence is available at GenBank (accession number - MG651603). FASTQ file is also available in the Sequence Read Archive (SRA) repository with accession number SRR6363036. Resultsi Identification of an episome in C. Violaceum strain ATCC 1242. While extracting genomic DNA
from C. violaceum strain ATCC 12472 to construct a genomic library, we noticed after agarose gel electrophoresis
the recurrence of a DNA species smaller than expected for high molecular weight genomic DNA in our prepa-
rations. We hypothesized that this DNA species could be a circular episome. We therefore carried out standard SCIENTIFIC Reports | (2018) 8:5327 | DOI:10.1038/s41598-018-23708-5 2 www.nature.com/scientificreports/ Figure 2. Restriction digestion of extra-chromosomal DNA extracted from eight different C. violaceum strains. Samples were digested with KpnI, BamHI and EcoRI for one hour at 37 °C. “−” reflects non digested samples
and L is DNA ladder. 0.8% TBE agarose gel stained with ethidium bromide. Figure 2. Restriction digestion of extra-chromosomal DNA extracted from eight different C. violaceum strains. Samples were digested with KpnI, BamHI and EcoRI for one hour at 37 °C. “−” reflects non digested samples
and L is DNA ladder. 0.8% TBE agarose gel stained with ethidium bromide. Figure 3. Kymographs showing the extensions of circular and nicked forms of the episome in nanofluidic channels. Competitive binding was used in order to produce the emission intensity pattern along the linear for of the plasmid. Figure 3. Kymographs showing the extensions of circular and nicked forms of the episome in nanofluidic channels. Competitive binding was used in order to produce the emission intensity pattern along the linear for of the plasmid. plasmid DNA preparations and analyzed the purified DNA by agarose gel electrophoresis and ethidium bromide
staining. As can be seen in lane 2 of Fig. 1, the preparation contained contaminating high molecular weight
genomic DNA trapped in the well but also a species with mobility much greater than 10 kb, our putative episome plasmid DNA preparations and analyzed the purified DNA by agarose gel electrophoresis and ethidium bromide
staining. As can be seen in lane 2 of Fig. 1, the preparation contained contaminating high molecular weight
genomic DNA trapped in the well but also a species with mobility much greater than 10 kb, our putative episome plasmid DNA preparations and analyzed the purified DNA by agarose gel electrophoresis and ethidium bromide
staining. As can be seen in lane 2 of Fig. www.nature.com/scientificreports/ www.nature.com/scientificreports/ re.com/scientificreports/
as indicated by a star symbol A third faster migrating species was also observed We next performed a restriction
Lenght, bp
44,212
G + C content
65.96%
Total ORFs
39
Percentage of plasmid sequence constituting coding regions
89.66%
Average ORF lenght, bp
1017
Number of conserved hypothetical proteins
28
Number of hypothetical proteins
1
Table 1. General features of pChV1. Figure 4. Map of the pChV1 plasmid. Most of the phage-related genes are present in the same region. The
distribution of the plasmid partitioning genes is in accordance to what is seen in the literature. Blue, red and
green arrows depict phage, plasmid and hypothetical ORFS respectively. Lenght, bp
44,212
G + C content
65.96%
Total ORFs
39
Percentage of plasmid sequence constituting coding regions
89.66%
Average ORF lenght, bp
1017
Number of conserved hypothetical proteins
28
Number of hypothetical proteins
1
Table 1. General features of pChV1. Figure 4. Map of the pChV1 plasmid. Most of the phage-related genes are present in the same region. The
distribution of the plasmid partitioning genes is in accordance to what is seen in the literature. Blue, red and
green arrows depict phage, plasmid and hypothetical ORFS respectively. Figure 4. Map of the pChV1 plasmid. Most of the phage-related genes are present in the same region. The
distribution of the plasmid partitioning genes is in accordance to what is seen in the literature. Blue, red and
green arrows depict phage, plasmid and hypothetical ORFS respectively. as indicated by a star symbol. A third faster migrating species was also observed. We next performed a restriction
enzyme analysis of our preparation with KpnI, BamHI or EcoRI (Fig. 1 lanes 3–5). Consistent with lineariza-
tion of a circular DNA molecule, digestion with KpnI resulted in a single band and disappearance of genomic
DNA, both the DNA trapped in the well and the third species described above, due to characteristic smearing of
genomic DNA digestions (Fig. 1 lane 3). Digestion with BamHI or EcoRI resulted instead in defined patterns of
discrete DNA fragments (Fig. 1 lanes 4 and 5). We then performed the same analysis using seven different addi-
tional C. violaceum strains (Fig. 2). Preparations from strains CVAC02, CVAC05, and CVT8 appeared to contain
a putative episome similar to strain ATCC 12472, while preparations from strains CV026, CVT19 and CVRP5
appeared to contain only genomic DNA. Resultsi 1, the preparation contained contaminating high molecular weight
genomic DNA trapped in the well but also a species with mobility much greater than 10 kb, our putative episome SCIENTIFIC Reports | (2018) 8:5327 | DOI:10.1038/s41598-018-23708-5 3 www.nature.com/scientificreports/ The preparation from strain CVT24 also seemed to contain a putative
episome species but the result from the restriction analysis is difficult to interpret. Thus, we identified an episome
in C. violaceum strain ATCC 12472 and propose to name it pChV1. as indicated by a star symbol. A third faster migrating species was also observed. We next performed a restriction
enzyme analysis of our preparation with KpnI, BamHI or EcoRI (Fig. 1 lanes 3–5). Consistent with lineariza-
tion of a circular DNA molecule, digestion with KpnI resulted in a single band and disappearance of genomic
DNA, both the DNA trapped in the well and the third species described above, due to characteristic smearing of
genomic DNA digestions (Fig. 1 lane 3). Digestion with BamHI or EcoRI resulted instead in defined patterns of
discrete DNA fragments (Fig. 1 lanes 4 and 5). We then performed the same analysis using seven different addi-
tional C. violaceum strains (Fig. 2). Preparations from strains CVAC02, CVAC05, and CVT8 appeared to contain
a putative episome similar to strain ATCC 12472, while preparations from strains CV026, CVT19 and CVRP5
appeared to contain only genomic DNA. The preparation from strain CVT24 also seemed to contain a putative
episome species but the result from the restriction analysis is difficult to interpret. Thus, we identified an episome
in C. violaceum strain ATCC 12472 and propose to name it pChV1. Episome pChV1 is a circular plasmid. The above restriction enzyme analysis suggested that pChV1
is a circular DNA molecule. To verify this hypothesis the episome DNA was purified after gel electrophore-
sis (Fig. 1 lane 2, band indicated with a star symbol) and stained with YOYO-1 (a bis-intercalator fluorescent
dye) and Netropsin (a minor groove binder of AT-rich sequences of double-stranded DNA) that competes with
YOYO-1 intercalation19. Such stained preparations were diluted and injected in nanochannels to observe indi-
vidual extended episome DNA molecules by fluorescence microscopy (Fig. 3A). www.nature.com/scientificreports/ Initially, the contour length of SCIENTIFIC Reports | (2018) 8:5327 | DOI:10.1038/s41598-018-23708-5 4 www.nature.com/scientificreports/ ORF Number
Gene
Length
Domains
Best Hit
Identity
ORF_01
Chromosome partitioning protein ParA
840
ParA/Soj/Fer4 NifH
93%
ORF_02
Partitioning protein ParB
885
ParB
96%
ORF_03
Plasmid replication protein RepA
951
Plasmid replication initiator protein
49%
ORF_04
Conserved hypothetical protein
477
none
82%
ORF_05
Conserved hypothetical protein/putative bacteriophage
lysis protein
1029
COG4623
82%
ORF_06
Conserved hypothetical protein
312
SlyX
97%
ORF_07
Conserved hypothetical protein
756
Cadherin repeat/Ca2+ binding
32%
ORF_08
Conserved hypothetical protein
1422
none
44%
ORF_09
Conserved hypothetical protein
1887
none
60%
ORF_10
Conserved hypothetical protein
363
none
99%
ORF_11
Conserved hypothetical protein
219
none
95%
ORF_12
Conserved hypothetical protein
1167
DUF4157
48%
ORF_13
Toxin
801
RhsA
52%
ORF_14
Conserved hypothetical protein
708
none
63%
ORF_15
Conserved hypothetical protein
1125
none
98%
ORF_16
Conserved Hypothetical protein
126
none
52%
ORF_17
Conserved hypothetical protein
615
Transposase
99%
ORF_18
Hypothetical protein
726
none
ND
ORF_19
Conserved hypothetical protein
891
DUF4255
95%
ORF_20
Conserved hypothetical protein
1854
ATPase AAA domain
72%
ORF_21
Conserved hypothetical protein
2415
none
62%
ORF_22
Conserved hypothetical protein
453
none
78%
ORF_23
Conserved hypothetical protein
3768
DUF342
95%
ORF_24
Conserved hypothetical protein
2439
none
82%
ORF_25
Conserved hypothetical protein
993
ribonuclease e/dihydrolipoamide
succyniltransferase
66%
ORF_26
Conserved hypothetical protein
3168
TIGR02243(phage tail-like region)
77%
ORF_27
Conserved hypothetical protein
441
none
90%
ORF_28
Phage baseplate assembly protein W
399
GPW gp25
71%
ORF_29
Conserved hypothetical protein
1593
Phage Base V
59%
ORF_30
Conserved hypothetical protein
729
Phage Tube
41%
ORF_31
Conserved Hypothetical protein
165
none
52%
ORF_32
Conserved hypothetical protein
456
Phage T4 gp19
40%
ORF_33
Phage tail protein
447
Phage T4 gp19
91%
ORF_34
Phage tail sheath protein
1416
COG3497(phage tail sheath protein FI)
63%
ORF_35
DNA invertase
624
mpi/SR ResInv(Recombinase;DNA
binding)/HTH Hin like
97%
ORF_36
Conserved hypothetical protein
348
HTH_XRE (transcriptional regulator
family)/xenobiotic response
97%
ORF_37
Toxin HipA
1350
HipA/Rna Pol
90%
ORF_38
Conserved Hypothetical protein
126
none
52%
ORF_39
Plasmid replication initiator protein
1182
RPA
54%
T bl 2 Li t f ORF f
d i
ChV1 Table 2. List of ORFs found in pChV1. individual episome DNA molecules averaged circa 6 μm (Fig. 3A - left kymograph). However, after prolonged
illumination, the accumulation of nicks in the DNA molecule induced double strand breakage and linearization
of the circular plasmid, evidenced by an increase in contour length (Fig. 3B - right kymograph). www.nature.com/scientificreports/ www.nature.com/scientificreports/ Figure 5. GC content profile of pChV1. Two points of low GC content are observable in the 1) beginning-end
of the plasmid sequence and in the 2) 27000 bp region (second chart, below). Figure 5. GC content profile of pChV1. Two points of low GC content are observable in the 1) beginning-end
of the plasmid sequence and in the 2) 27000 bp region (second chart, below). Plasmid maintenance genes. The plasmid has at least 4 known genes related to plasmid segrega-
tion/replication: parA, parB, repA and a gene with RPA domain, involved in plasmid replication initiation
(Table 2). parA and parB encode the ParA and ParB proteins, respectively. These proteins are part of the Type
I plasmid-partitioning system and are responsible for ensuring the correct propagation of plasmids to daughter
cells throughout cell division24. This partitioning system is founded in prophages, plasmids and chromosomes25. RepA is a protein related to plasmid replication and is characteristic of P1 plasmids. Structural phage genes. An abundant number of genes related to phage structure are present in the
sequence of the plasmid. Genes that codify the baseplate, sheath and tail proteins as well as conserved hypotheti-
cal genes with domains related to phage structure are in close proximity in the pChV1 sequence. Other genes. A DNA invertase (ORF_35), an enzyme that catalyzes site-specific recombination in phages
was found. A conserved hypothetical protein (ORF_17) with a transposase domain is also present in the plas-
mid sequence. Toxins (ORF_13 and ORF_37) that may be related to the toxin-antitoxin (TA) system respon-
sible for assuring the survival only for the cells with a copy of the lisogenyzed phage were also located. Other
worthy-mention genes are: conserved hypothetical proteins with Ribonuclease E domain, XRE domain and
ATPase AAA domain. GC profile and GC-skew. We were able to identify two points in the sequence of pChV1 where the GC con-
tent drops when comparing to the whole sequence (Fig. 5). These variations also qualitatively agree with intensity
variaitons in the single molecules studies in Fig. 3. This might reflect the presence of two origins of replication
that are present in P1-like plasmids, oriR and oriL. GC-skew also helps predicting the location of the leading and
lagging strand and cumulative GC-skew values reflect the origin and terminus points of replication26. www.nature.com/scientificreports/ Lambda phage
DNA (48,502 bp) was used as an internal standard to convert extension from pixels to basepairs. Using a scal-
ing factor of 1.8423 for going from circular to linear extension, we estimated that the longest episome molecule
detected was circa 44 kb in size. DNA barcode analysis revealed mostly GC-rich regions with two AT-rich regions
of darker signal. individual episome DNA molecules averaged circa 6 μm (Fig. 3A - left kymograph). However, after prolonged
illumination, the accumulation of nicks in the DNA molecule induced double strand breakage and linearization
of the circular plasmid, evidenced by an increase in contour length (Fig. 3B - right kymograph). Lambda phage
DNA (48,502 bp) was used as an internal standard to convert extension from pixels to basepairs. Using a scal-
ing factor of 1.8423 for going from circular to linear extension, we estimated that the longest episome molecule
detected was circa 44 kb in size. DNA barcode analysis revealed mostly GC-rich regions with two AT-rich regions
of darker signal. The pChV1 DNA sequence. The complete sequence of pChV1 revealed a circular element with 44,212 bp
with a G + C content of 65.96% (Table 1). 39 Open Reading Frames (ORFs) were found, which comprises 89,66%
of the whole plasmid (Fig. 4). From these, 28 are conserved hypothetical proteins and 1 is a hypothetical protein. Comparing the ORFs of the plasmid with other organisms, we observed that 17 (43%) of the ORFs have similarity
with ORFs from Pseudogulbenkiania ferrooxidans. No tRNAs genes were found. We also searched for homology
with bacteriophages and the BLAST analysis did not give any similarity with any phage genomes. SCIENTIFIC Reports | (2018) 8:5327 | DOI:10.1038/s41598-018-23708-5 5 www.nature.com/scientificreports/ www.nature.com/scientificreports/ tificreports/
Figure 6. GC-skew of pChV1. The cumulative GC-skew (red curve) has two decline points, characteristic of
origins of replication and might reflect oriR and oriL. Figure 6. GC-skew of pChV1. The cumulative GC-skew (red curve) has two decline points, characteristic o
origins of replication and might reflect oriR and oriL. Figure 6. GC skew of pChV1. The cumulative GC skew (red curve) has two decline points, characteristic of
origins of replication and might reflect oriR and oriL. Figure 7. Schematic chart showing the genomic context in which the 19 bp inverted repeated sequence are
flanking the partitioning-related genes parA and parB. The diagram is not to scale. Figure 7. Schematic chart showing the genomic context in which the 19 bp inverted repeated sequence are
flanking the partitioning-related genes parA and parB. The diagram is not to scale. Coordinates
Strand
Sequence
Type of sequence
42355–42373
44177–44159
+
+
TGTAGCAAGTTGCTACACT
ACATCGTTCAACGATGTGA
Inverted repeat
sequence
5113–5120
10723–10730
+
+
AAATATTT
Palindrome
44161–44177
42355–42371
−
+
TGTAGCAAg/cTTGCTACA
Palindrome
27010–27016
26990–26996
−
+
ATAt/aTAT
Palindrome
1608–1615
32309–32316
+
+
TGAATTCA
Palindrome
1465–1478
41121–41134
+
−
TTTTTAACTAAAAA
TTTATAGTTAAGAA
Palindrome
304–312
132–140
−
+
TTGAt/aTCAA
Palindrome
34202–34209
33338–33345
+
−
AATTAATT
AAGTAATT
Palindrome
34414–34424
4595–4605
+
−
ATTTGTCATAT
Palindrome
27884–27891
32052–32059
−
−
TATTCATA
Palindrome
Table 3. Pairs of Inverted repeated and palindromic sequences founded in pChV1. Coordinates
Strand
Sequence
Type of sequence
42355–42373
44177–44159
+
+
TGTAGCAAGTTGCTACACT
ACATCGTTCAACGATGTGA
Inverted repeat
sequence
5113–5120
10723–10730
+
+
AAATATTT
Palindrome
44161–44177
42355–42371
−
+
TGTAGCAAg/cTTGCTACA
Palindrome
27010–27016
26990–26996
−
+
ATAt/aTAT
Palindrome
1608–1615
32309–32316
+
+
TGAATTCA
Palindrome
1465–1478
41121–41134
+
−
TTTTTAACTAAAAA
TTTATAGTTAAGAA
Palindrome
304–312
132–140
−
+
TTGAt/aTCAA
Palindrome
34202–34209
33338–33345
+
−
AATTAATT
AAGTAATT
Palindrome
34414–34424
4595–4605
+
−
ATTTGTCATAT
Palindrome
27884–27891
32052–32059
−
−
TATTCATA
Palindrome
Table 3. Pairs of Inverted repeated and palindromic sequences founded in pChV1. Table 3. Pairs of Inverted repeated and palindromic sequences founded in pChV1. www.nature.com/scientificreports/ In our anal-
ysis, we can observe throughout the cumulative GC-skew curve, two regions that we could call minimum points
that sign the origins of replication oriR and oriL (Fig. 6). Repeated and palindromic sequences. A 19 bp inverted repeated sequence separated by 1,785 bp was
also located and may be involved in the circularization of the phage or other homologous recombination-based
process (Table 3). This pair of sequences is located flanking the partitioning related genes parA and parB (Fig. 7). Other inverted repeat sequences with size varying from 23 to 54 bp were also founded although the complemen-
tarity between the pair of repeats was not 100% (data not shown). Palindromic sequences located at two distinct
sites in the sequence and varying from 7 to 17 bp are also present (Table 3). SCIENTIFIC Reports | (2018) 8:5327 | DOI:10.1038/s41598-018-23708-5 6 www.nature.com/scientificreports/ www.nature.com/scientificreports/ After the sequencing of C. violaceum7, the presence of four different sequences of prophages (CvP1-4) were
observed in the C. violaceum’s genome28. Neither of these is related to the plasmid we report here. Before this,
tail-like particles were observed in C. violaceum by electron microscopy although no biological activity was asso-
ciated to them29,30. According to the sequence data and annotation, the plasmid founded in C. violaceum could be a P1-like virus
due to the presence of genes that encode for structural viral particles. Moreover, genes related to plasmid par-
titioning and the plasmid initiator protein RepA are strong evidence to classify this plasmid as a P1-like phage. Another hallmark of P1-like phages is the presence of toxin-antitoxin genes that constitute a plasmid addiction
system. In pChV1 two ORFs are predicted to be toxin genes (ORFs 13 and 37 with 52% and 90% of identity,
respectively) although further studies need to be done to confirm the presence of this system.f i
From our search for homology, we observed that pChV1 has a nucleotide sequence very different from other
phages described so far. This feature hampers the search for phage-related sequences, such as lox sites, incC and
incA and others, which are, in general, well conserved between other viruses, but does not exclude the existence of
them in pChV1. However, repeated sequences that are founded amongst other phages are also present in pChV1,
such as the 19 bp inverted repeated sequence (Fig. 7). p
p
q
g
Origins of replication are GC-poor regions and locating them in the plasmid may suggest the locals where
replication starts. Although we were not able to predict specific sequences that would correspond to origins of
replications in pChV1, the GC content profile and GC-skew showed two regions that might reflect oriR and oriL. oriR, that is used during plasmid maintenance replication, is in the same region as the parA, parB and repA genes. This co-location of a possible origin of replication and the plasmid maintenance genes is observable in pChV1. Conversely, oriL is related to lytic growth and is separated about 9 kb from oriR in P131. We suggest that the
second possible origin of replication founded in pChV1 (located approximately at 27 kbp) corresponds to oriL. www.nature.com/scientificreports/ p
g
p
p
pp
y
p
p
One notable feature is that when we aligned the predicted open reading frames using BLASTn (that searches a
nucleotide query in a nucleotide database) we obtained no significant result. Conversely, when BLASTx was used
(searches a translated nucleotide query in a protein database) we were able to identify genes with high degree of
confidence. This means that during evolution this virus accumulated many mutations on its DNA sequence but
conserved – to some extent - the amino acid composition of its proteins. For example, pChV1 has many ORFs
with more than 90% of identity with other genes found in bacteria (Table 2). When we aligned these same ORFs
using BLASTn we did not obtain any significant result. g
y
gi
Besides the presence of phage-related genes and sequences, some essential elements that would make pChV1
a functional P1-phage are still missing31. By the lack of evidence, we cannot conclude if this plasmid is a temper-
ate P1-like phage, or if it is a chimeric DNA, part bacteriophage or plasmid. Moreover, it could be a fragment
of DNA that is maintained inside C. violaceum by addiction systems but defective in its capacity of lisogeny. Conversely, the tail-like particles observed in the 1970s29,30 could be an evidence that, under stress, the phage
proteins encoded by pChV1 would be produced. p
y p
p
Genetic mobile elements are still important in the field of molecular biology. Beside this, the use of
phage-derived systems as tools has allowed genome manipulation of all kind of organisms. In this way, further
study of pChV1 would bring new ways to investigate genetic aspects of C. violaceum and maybe other species. Finally, pChV1 with its great number of hypothetical ORFs, is a rich reservoir of unexplored genes that might
contribute to our understanding of the mechanisms underlying viral infections and plasmids. Conclusion In our work, we discovered an extra-chromosomal DNA – that we named pChV1 - in the opportunistic pathogen
Chromobacterium violaceum. This plasmid is present as a low-copy plasmid and has most of its genetic apparatus
composed of ORFs with unknown function, making pChV1 an important source of genes to be further explored. More than this, when its biology is better understood, this element can be used in genetic studies in C. violaceum
as well as in other organisms. References 1. Garrity, G. M. & Holt, J. G. In Bergeys Manual of Systematic Bacteriology The Archaea and the Deeply Branching and Phototrophi
Bacteria 119–166, https://doi.org/10.1007/978-0-387-21609-6_15 (2001). 2. Durán, N. et al. MINIREVIEW Violacein: properties and biological activities. Biotechnol. Appl. Biochem. 133, 127–133 (2007). 3. Durán, M., Faljoni-Alario, A. & Durán, N. Chromobacterium violaceum and its important metabolites–review. Folia Microbio
(Praha). 55, 535–47 (2010). 2. Durán, N. et al. MINIREVIEW Violacein: properties and biological activities. Biotechnol. Appl. Biochem. 133, 127–133 (2007). 3 Durán M Faljoni Alario A & Durán N Chromobacterium violaceum and its important metabolites review Folia Microbiol 2. Durán, N. et al. MINIREVIEW Violacein: properties and biological activities. Biotechnol. Appl. Biochem. 133, 127–133 (2007). 3. Durán, M., Faljoni-Alario, A. & Durán, N. Chromobacterium violaceum and its important metabolites–review. Folia Microbiol. (Praha) 55 535 47 (2010) (
)
(
)
4. Durán, M. et al. Potential applications of violacein: A microbial pigment. Med. Chem. Res. 21, 1524–1532 (2012). 4. Durán, M. et al. Potential applications of violacein: A microbial pigment. Med. Chem. Res. 21, 1524–1532 (2012). 5. Yang, C.-H. & Li, Y.-H. Chromobacterium violaceum infection: a clinical review of an important but neglected infection. J. Med. Assoc. 74, 435–41 (2011). 6. Luz, K. G. et al. Chromobacterium violaceum: a fatal case in the northeast of the Brazil. J Bras Patol Med Lab 50, 278–279 (2014).h 6. Luz, K. G. et al. Chromobacterium violaceum: a fatal case in the northeast of the Brazil. J Bras Patol Med Lab 50, 278–279 (2014). l
A T
l Th
l
f Ch
b
l
l
k bl
d
l
bl 6. Luz, K. G. et al. Chromobacterium violaceum: a fatal case in the northeast of the Brazil. J Bras Patol Med Lab 50, 278 279 (2014). 7. Vasconcelos, A. T. R. et al. The complete genome sequence of Chromobacterium violaceum reveals remarkable and exploitabl
bacterial adaptability. Proc. Natl. Acad. Sci. USA 100, 11660–5 (2003).f p
y
8. Baraúna, R. a. et al. Proteomics Analysis of the Effects of Cyanate on Chromobacterium violaceum Metabolism. Genes (Basel). 2,
(736–747 (2011).hli p
y
8. Baraúna, R. a. et al. Proteomics Analysis of the Effects of Cyanate on Chromobacterium violaceum Metabolism. Genes (Basel). 2
(736–747 (2011). l Th
fl
f
h
fil
f
h
b
l
b l
(
) file of Chromobacterium violaceum. BMC Microbiol. 14, 267 (2014 9. Lima, D. C. et al. Discussionhi The first bacteriophage was discovered in the 1950’s27 and since then, the number and variety of new viruses that
infect bacteria has grown considerably, reaching more than 1,300 genome projects according to the NCBI data-
base. While researching the opportunistic pathogen C. violaceum, in genomic preparations we observed an
extra-chromosomal DNA of high molecular weight (but lower than it would be if it was genomic DNA). We then
isolated and sequenced this putative plasmid which proved to have genes from the P1 bacteriophage/plasmid group. SCIENTIFIC Reports | (2018) 8:5327 | DOI:10.1038/s41598-018-23708-5 7 p
g
2. Durán, N. et al. MINIREVIEW Violacein: properties and biological activities. Biotechnol. Appl. Biochem. 133, 127–133 (2007).
3. Durán, M., Faljoni-Alario, A. & Durán, N. Chromobacterium violaceum and its important metabolites–review. Folia Microbiol.
(P
h ) 55 535 47 (2010) Acknowledgementsh g
The authors would like to thank UFRN, CNPq, CAPES for financial support. The authors are also grateful to
Mauro Modesti who performed Restriction Digestion assay and also hosted D.C.L. in his lab. www.nature.com/scientificreports/ Th
f l
d
l
d
b l 24. Salje, J. Plasmid segregation: how to survive as an extra piece of DNA. Crit. Rev. Biochem. Mol. Biol. 45, 296 317 (2010). 25. Pinto, U. M., Pappas, K. M. & Winans, S. C. The ABCs of plasmid replication and segregation. Nat. Rev. Microbiol. 10, 755–765
(2012). 6. Eppinger, M., Baar, C., Raddatz, G., Huson, D. H. & Schuster, S. C. Comparative analysis of four Campylobacterales. Nat. Rev
Microbiol. 2, 872–885 (2004).h 27. Bertani, G. Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J. Bacteriol. 62, 293–300 (1951). l
d
l
h
d
f
h
b
l
T
l 27. Bertani, G. Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J. Bacteriol. 62, 293–300 (1951). 28 De Almeida, R et al Bacteriophages and insertion sequences of Chromobacterium violaceum ATCC 12472 Genet Mol Res 3, Bertani, G. Studies on lysogenesis. I. The mode of phage liberation b 27. Bertani, G. Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J. Bacteriol. 62, 293–300 (1951). 28. De Almeida, R. et al. Bacteriophages and insertion sequences of Chromobacterium violaceum ATCC 12472. Genet. Mol. Res. 3,
76–84 (2004). 27. Bertani, G. Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J. Bacteriol. 62, 293 300 (195
28. De Almeida, R. et al. Bacteriophages and insertion sequences of Chromobacterium violaceum ATCC 12472. Genet. Mol. R
76–84 (2004). 29. Rucinsky, T. E., Gregory, J. P. & Cota-Robles, E. H. Organization of bacteriophage tail-like particles in cells of Chromobacterium
violaceum. J. Bacteriol. 110, 754–757 (1972).h 30. Rucinsky, T. E. & Cota-Robles, E. H. The intracellular organization of bacteriophage tail-like particles in cells of Chromobacterium
violaceum following mitomycin C treatment. J. Ultrastruct. Res. 43, 260–269 (1973). 30. Rucinsky, T. E. & Cota-Robles, E. H. The intracellular organization of bacteriophage tail-like particles in cells of Chromobacterium
violaceum following mitomycin C treatment. J. Ultrastruct. Res. 43, 260–269 (1973). h
violaceum following mitomycin C treatment. J. Ultrastruct. Res 31. Lobocka, M. B. et al. Genome of Bacteriophage P1. 186 (2004). Author Contributions D.C.L. participated in the design of the study, acquisition of data and analysis and interpretation of data and
drafted the manuscript. L.K.N. and F.W. performed optical mapping analysis of DNA and interpreted the results. S.R.B.M. contributed to the study conception and design, writing of the manuscript and overall supervision. All
authors read and approved the final manuscript. www.nature.com/scientificreports/ www.nature.com/scientificreports/ 14. Skogman, M. E., Kanerva, S., Manner, S., Vuorela, P. M. & Fallarero, A. Flavones as quorum sensing inhibitors identified by a n
optimized screening platform using chromobacterium violaceum as reporter bacteria. Molecules 21 (2016). 14. Skogman, M. E., Kanerva, S., Manner, S., Vuorela, P. M. & Fallarero, A. Flavones as quorum sensing inhibitors identified by a newly
optimized screening platform using chromobacterium violaceum as reporter bacteria. Molecules 21 (2016). p
g p
g
p
15. Ciprandi, A. et al. Proteomic Response to Arsenic Stress in Chromobacterium violaceum. J. Integr. OMICS 2, 69–73 (2012). p
p
g
16. Gene, A., An, T., Lecula, M. O., Tech, B. I. O. & Broetto, N. L. Stable transformation of Chromobacterium violaceum with a br
host-range plasmid. Biologia (Bratisl). 450–454, https://doi.org/10.1007/s00253-005-0140-5 (2006). g p
g
p
g
7. da Silva Neto, J. F., Negretto, C. C. & Netto, L. E. S. Analysis of the organic hydroperoxide response of Chromobacterium violaceum
reveals that OhrR is a cys-based redox sensor regulated by thioredoxin. PLoS One 7, e47090 (2012). g p
g
p
g
17. da Silva Neto, J. F., Negretto, C. C. & Netto, L. E. S. Analysis of the organic hydroperoxide response o
reveals that OhrR is a cys-based redox sensor regulated by thioredoxin. PLoS One 7, e47090 (2012). y
g
y
8. Nyberg, L. K. et al. Rapid identification of intact bacterial resistance plasmids via optical mapping of single DNA molecules. Sci. Rep
6 (2016).i 19. Nilsson, A. N. et al. Competitive binding-based optical DNA mapping for fast identification of Bacteria - Multi-ligand tra
matrix theory and experimental applications on Escherichia coli. Nucleic Acids Res. 42 (2014). y
p
pp
20. Persson, F. & Tegenfeldt, J. O. DNA in nanochannels—directly visualizing genomic information. Chem. Soc. Rev. 39, 985 (2010 1. Bailey, T. L. & Elkan, C. Fitting a mixture model by expectation maximization to discover motifs in biopolymers. Proc. Int. Conf
Intell. Syst. Mol. Biol. 2, 28–36 (1994).i y
2. Gao, F. & Zhang, C. T. GC-Profile: A web-based tool for visualizing and analyzing the variation of GC content in genomic sequences
Nucleic Acids Res. 34 (2006). (
)
23. Alizadehheidari, M. et al. Nanoconfined circular and linear DNA: Equilibrium conformations and unfolding kinetic
Macromolecules 48, 871–878 (2015). 24. Salje, J. Plasmid segregation: how to survive as an extra piece of DNA. Crit. Rev. Biochem. Mol. Biol. 45, 296–317 (2010). References The influence of iron on the proteomic profile of Chromobacterium violaceum. BMC Microbiol. 14, 267 (2014). 0. Castro, D. et al. Proteomic analysis of Chromobacterium violaceum and its adaptability to stress Proteomic analysis o
Chromobacterium violaceum and its adaptability to stress. BMC Microbiol. 15 (2015). ,hl
p
pi
,
(
)
10. Castro, D. et al. Proteomic analysis of Chromobacterium violaceum and its adaptability to stress Proteomic analysis of
Chromobacterium violaceum and its adaptability to stress. BMC Microbiol. 15 (2015). 1. Duarte, F. T. et al. GeLC-MS-based proteomics of Chromobacterium violaceum: comparison of proteome changes elicited by
hydrogen peroxide. (2016). hydrogen peroxide. (2016). 12. Zhu, H., He, C.-C. & Chu, Q.-H. Inhibition of quorum sensing in Chromobacterium violaceum by pigments extracted from
A
i
l
i
i
l
L
A
l Mi
bi l 1 6 h
//d i
/10 1111/j 1472 765X 2011 02993
(2011) y
g
2. Zhu, H., He, C.-C. & Chu, Q.-H. Inhibition of quorum sensing in Chromobacterium violaceum by pigments extracted from
Auricularia auricular. Lett. Appl. Microbiol. 1–6, https://doi.org/10.1111/j.1472-765X.2011.02993.x (2011).f pp
,
p
g
j
(
)
13. Chaudhari, V., Gosai, H., Raval, S. & Kothari, V. Effect of certain natural products and organic solvents on quorum sensing in
Chromobacterium violaceum. Asian Pac. J. Trop. Med. 7, S204–S211 (2014). pp
p
g
j
(
)
13. Chaudhari, V., Gosai, H., Raval, S. & Kothari, V. Effect of certain natural products and organic solvents on quorum sensing in
Chromobacterium violaceum. Asian Pac. J. Trop. Med. 7, S204–S211 (2014). SCIENTIFIC Reports | (2018) 8:5327 | DOI:10.1038/s41598-018-23708-5 8 SCIENTIFIC Reports | (2018) 8:5327 | DOI:10.1038/s41598-018-23708-5 Additional Informationh Competing Interests: The authors declare no competing interests. Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and
institutional affiliations. Open Access This article is licensed under a Creative Commons Attribution 4.0 International
License, which permits use, sharing, adaptation, distribution and reproduction in any medium or
format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Cre-
ative Commons license, and indicate if changes were made. The images or other third party material in this
article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the
material. If material is not included in the article’s Creative Commons license and your intended use is not per-
mitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the
copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. © The Author(s) 2018 SCIENTIFIC Reports | (2018) 8:5327 | DOI:10.1038/s41598-018-23708-5 9
|
https://openalex.org/W2043861527
|
https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1004284&type=printable
|
English
| null |
The Sequence-Specific Transcription Factor c-Jun Targets Cockayne Syndrome Protein B to Regulate Transcription and Chromatin Structure
|
PLOS genetics
| 2,014
|
cc-by
| 14,943
|
Abstract The funders had no role in study
analysis, decision to publish, or preparation of the manuscript. supported by grants from NIH; GM 084983 (HYF) and DK098769 (KJW). The funders had no role in study design, data collection and
ish, or preparation of the manuscript. Funding: This work was supported by grants from NIH; GM 084983 (HYF) and DK098769
analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. * E-mail: hfan@mail.med.upenn.edu The Sequence-Specific Transcription Factor c-Jun Targets
Cockayne Syndrome Protein B to Regulate Transcription
and Chromatin Structure Robert J. Lake1,2, Erica L. Boetefuer1,2,3, Pei-Fang Tsai1,2, Jieun Jeong1,4,5, Inchan Choi4,5,
Kyoung-Jae Won1,4,5, Hua-Ying Fan1,2,4,5* 1 Epigenetics Program, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America, 2 Department of Biochemistry and
Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America, 3 Biology Graduate Program, Graduate School
of Arts and Sciences, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America, 4 Institute for Diabetes Obesity and Metabolism, Perelman School of
Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America, 5 Department of Genetics, Perelman School of Medicine, University of
Pennsylvania, Philadelphia, Pennsylvania, United States of America April 2014 | Volume 10 | Issue 4 | e1004284 PLOS Genetics | www.plosgenetics.org Abstract Cockayne syndrome is an inherited premature aging disease associated with numerous developmental and neurological
defects, and mutations in the gene encoding the CSB protein account for the majority of Cockayne syndrome cases. Accumulating evidence suggests that CSB functions in transcription regulation, in addition to its roles in DNA repair, and
those defects in this transcriptional activity might contribute to the clinical features of Cockayne syndrome. Transcription
profiling studies have so far uncovered CSB-dependent effects on gene expression; however, the direct targets of CSB’s
transcriptional activity remain largely unknown. In this paper, we report the first comprehensive analysis of CSB genomic
occupancy during replicative cell growth. We found that CSB occupancy sites display a high correlation to regions with
epigenetic features of promoters and enhancers. Furthermore, we found that CSB occupancy is enriched at sites containing
the TPA-response element. Consistent with this binding site preference, we show that CSB and the transcription factor c-Jun
can be found in the same protein-DNA complex, suggesting that c-Jun can target CSB to specific genomic regions. In
support of this notion, we observed decreased CSB occupancy of TPA-response elements when c-Jun levels were
diminished. By modulating CSB abundance, we found that CSB can influence the expression of nearby genes and impact
nucleosome positioning in the vicinity of its binding site. These results indicate that CSB can be targeted to specific genomic
loci by sequence-specific transcription factors to regulate transcription and local chromatin structure. Additionally,
comparison of CSB occupancy sites with the MSigDB Pathways database suggests that CSB might function in peroxisome
proliferation, EGF receptor transactivation, G protein signaling and NF-kB activation, shedding new light on the possible
causes and mechanisms of Cockayne syndrome. Citation: Lake RJ, Boetefuer EL, Tsai P-F, Jeong J, Choi I, et al. (2014) The Sequence-Specific Transcription Factor c-Jun Targets Cockayne Syndrome Protein B to
Regulate Transcription and Chromatin Structure. PLoS Genet 10(4): e1004284. doi:10.1371/journal.pgen.1004284 Editor: Alan M. Weiner, University of Washington, United States of America Received September 27, 2013; Accepted February 20, 2014; Published April 17, 2014 Copyright: 2014 Lake et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work was supported by grants from NIH; GM 084983 (HYF) and DK098769 (KJW). Citation: Lake RJ, Boetefuer EL, Tsai P-F, Jeong J, Choi I, et al. (2014) The Sequence-Specific Transcription Factor c-Jun Targets Cocka
Regulate Transcription and Chromatin Structure. PLoS Genet 10(4): e1004284. doi:10.1371/journal.pgen.1004284 The genomic occupancy of CSB To do this, we compared signal intensities for each
CSB peak identified by HOMER to the CSBDN1 signal intensity,
regardless of whether the CSBDN1 signal was identified as a peak
by HOMER. Similarly, we compared signal intensities for each
CSBDN1 peak identified by HOMER to the CSB signal intensity,
regardless of whether the CSB signal was identified as a peak by
HOMER. Signal intensities were compared over a 200 bp region. If the difference between signal intensities was 4-fold or greater
and the p-value for that difference was #0.0001, the signal was
classified as unique; the remaining signals were classified as
common (Tables S1, S2). Among them, we identified 6,398 peaks
unique to CSB and 877 peaks unique to CSBDN1 (Table 1). As
shown in Figure 1B, ,36% of the CSB occupancy sites were
unique to CSB and ,24% of the CSBDN1 occupancy sites were
unique to CSBDN1. CSB is best known for its function in transcription-coupled
DNA repair, a process that preferentially removes bulky DNA
lesions that stall transcription, such as those created by UV
irradiation [18,19]. CSB is one of the first proteins recruited to
sites of lesion-stalled transcription, and the ability of CSB to
hydrolyze ATP is essential for this association [20]. After its arrival
to
lesion-stalled
transcription,
CSB
appears
to
have
both
chromatin remodeling-dependent and remodeling-independent
functions. One function of CSB is to recruit the DNA repair
machinery [20], and this activity does not rely upon nucleosome
repositioning by CSB [8]. The chromatin remodeling activity of
CSB, on the other hand, is likely to be important for regulating
protein DNA associations or nucleosome positioning necessary for
the repair process or the resumption of transcription after repair. CSB is also involved in the repair of oxidative lesions in both
nuclear and mitochondrial DNA [21–24]. We then classified the CSB and CSBDN1 occupancy sites into
seven functional categories (see Materials and Methods), using the
UCSC RefSeq gene annotations and the CEAS package [32]. The
distribution of CSB peaks among those categories was largely
consistent with a random distribution (Figure S2); however, we
observed modest enrichment of CSB occupancy at promoter
regions (1.5% for CSB peaks vs. 1.1% for the genomic distribution,
binomial test p-value of 7.3e-07) and modest depletion at 39UTRs
(1.1% for CSB peaks vs. 1.4% for the genomic distribution,
binomial test p-value of 1.6e-05). The genomic occupancy of CSB To identify genomic regions that CSB occupies, we performed
chromatin immunoprecipitation using a monoclonal antibody
raised against CSB, followed by deep sequencing. CS1AN-sv cells
that were reconstituted with CSB were used for these experiments,
as CS1AN-sv cells do not express the alternatively spliced CSB-
PiggyBac fusion protein (Figure S1) [30]; this cell line is
hemizygous for the CSB locus and the retained CSB allele has a
premature stop codon at amino acid 337 [19] (see Materials and
Methods). Additionally, to understand the importance of CSB’s
nucleosome-remodeling activity in CSB function, we also included
in these assays the remodeling-defective CSBDN1 derivative
(Figure 1A) [8]. The resulting sequencing reads were mapped to the human
genome (HG19 assembly) using the Bowtie aligner [31]. Peaks
were identified using HOMER (Hypergeometric Optimization of
Motif EnRichment) with a default option (FDR = 0.001 and
Poisson p-value cut-off = 0.0001) on ChIPed samples against
matching input samples. In total, we recovered 17,779 CSB peaks
and 3,607 CSBDN1 peaks (Table 1). quite diverse [2]. For example, CSB has been shown to alter DNA
conformation and to actively wrap DNA [13,14], both in an ATP-
dependent manner. Additionally, CSB has been shown to use the
energy from ATP hydrolysis to alter chromatin structure [8,13]. Most recently, the NAP1-like histone chaperone was shown to
significantly enhance the chromatin remodeling activity of CSB
and, together, these two proteins can centralize mononucleosomes,
suggesting a potential function of CSB in nucleosome spacing
[8,15,16]. Several ATP-independent activities have also been
ascribed to CSB. Such activities include dissociating non-histone
proteins from DNA, annealing single-stranded DNA, and mod-
ulating the activities of DNA repair proteins [17]. quite diverse [2]. For example, CSB has been shown to alter DNA
conformation and to actively wrap DNA [13,14], both in an ATP-
dependent manner. Additionally, CSB has been shown to use the
energy from ATP hydrolysis to alter chromatin structure [8,13]. Most recently, the NAP1-like histone chaperone was shown to
significantly enhance the chromatin remodeling activity of CSB
and, together, these two proteins can centralize mononucleosomes,
suggesting a potential function of CSB in nucleosome spacing
[8,15,16]. Several ATP-independent activities have also been
ascribed to CSB. Such activities include dissociating non-histone
proteins from DNA, annealing single-stranded DNA, and mod-
ulating the activities of DNA repair proteins [17]. We subsequently classified peaks unique to CSB or CSBDN1
(Figure 1B). C-Jun Targets CSB to Regulate Transcription C-Jun Targets CSB to Regulate Transcription To understand better how CSB carries out its diverse functions
and to gain potential insights into the underlying mechanism of
Cockayne syndrome, we performed a genome-wide study of CSB
occupancy to elucidate the mechanisms of CSB targeting and to
understand the impacts of CSB occupancy on transcription
regulation. Author Summary Cockayne syndrome is a devastating inherited disease, in
which patients appear to age prematurely, have sun
sensitivity and suffer from profound neurological and
developmental
defects. Mutations
in
the
CSB
gene
account for the majority of Cockayne syndrome cases. CSB is an ATP-dependent chromatin remodeler, and these
proteins can use energy from ATP-hydrolysis to alter
contacts between DNA and histones of a nucleosome, the
basic units of chromatin structure. CSB functions in DNA
repair, but accumulating evidence reveals that CSB also
functions in transcription regulation. Here, we determined
the genomic localization of CSB to identify its gene targets
and found that CSB occupancy displays high correlation to
regions
with
epigenetic
features
of
promoters
and
enhancers. Furthermore, CSB is enriched at genomic
regions containing the binding site for the c-Jun tran-
scription factor, and we found that these two proteins
interact, uncovering a new targeting mechanism for CSB. We also demonstrate that CSB can influence gene
expression in the vicinity of its binding sites and alter
local chromatin structure. Together, this study supports
the hypothesis that defects in the regulation of gene
expression and chromatin structure by CSB might contrib-
ute to the diverse clinical features of Cockayne syndrome. Introduction protein factors to DNA. Additionally, some ATP-dependent
chromatin remodelers can assemble nucleosomes or create equally
spaced nucleosomes to facilitate the formation of higher-order
chromatin structure [5]. Most remodelers in isolation can alter
chromatin structure in vitro. The additional proteins that form
complexes with ATP-dependent chromatin remodelers are often
involved in enhancing the specific activity of the remodeler or
targeting the remodeling complex to specific genomic regions [6–
8]. Additionally, some SWI2/SNF2 family members can alter
contacts between DNA and non-histone proteins [9]. For example,
the MOT1 remodeler can use the energy from ATP hydrolysis to
dissociate TBP (TATA box binding protein) from DNA [9]. Cockayne syndrome is a devastating inherited disease in which
patients have features of premature aging, display increased sun
sensitivity, and suffer from profound neurological and develop-
mental defects [1]. Mutations in the gene encoding the CSB
(Cockayne syndrome complementation B) protein are associated
with the majority of Cockayne syndrome cases. CSB belongs to the
SWI2/SNF2
ATP-dependent
chromatin
remodeling
protein
family [2]. ATP-dependent chromatin remodelers are conserved
from yeast to human, and they are critical in regulating
fundamental nuclear processes, such as transcription and DNA
repair [3,4]. These proteins use ATP as energy to alter chromatin
structure non-covalently, resulting in changes in nucleosome
position, composition or conformation. By doing so, ATP-
dependent chromatin remodelers can regulate the access of In vitro, the CSB remodeler appears to interact with DNA in a
sequence-independent manner, and CSB displays both DNA and
nucleosome stimulated ATP hydrolysis activity [4,10–12]. The
biochemical activities that have been associated with CSB are April 2014 | Volume 10 | Issue 4 | e1004284 PLOS Genetics | www.plosgenetics.org 1 The genomic occupancy of CSB Interestingly, CSBDN1 peaks
displayed greater enrichment at promoter regions (3.1% for
CSBDN1 peaks vs. 1.1% for the genomic distribution, binomial
test p-value of 6.8e-23) and 59UTRs (1.1% for CSB peaks vs. 0.4%
for the genomic distribution, binomial test p-value of 1.4e-09)
(Figure S2). In addition to its well-documented function in DNA repair,
accumulating evidence indicates that CSB also participates in
transcription regulation [25–28]. CSB has been found to be in
complexes with both RNA polymerase II and RNA polymerase I. Interestingly, CSB does not appear to impact transcription in a
general manner, as transcription-profiling experiments revealed
that CSB has specific effects on gene expression. The results of that
study has lead to the intriguing hypothesis that Cockayne
syndrome might be, at least in part, a disease of transcription
deregulation [28,29]. Given that CSBDN1 is more significantly enriched at promoters
and 59 UTRs than CSB (p-values of 5.2e-11 and 1.8e-7,
respectively) (Figure S2), we calculated the number of CSB and
CSBDN1 peaks as a function of distance from transcription start April 2014 | Volume 10 | Issue 4 | e1004284 PLOS Genetics | www.plosgenetics.org 2 C-Jun Targets CSB to Regulate Transcription Figure 1. Overview of CSB and CSBDN1 ChIP-seq data. (A) Schematics of CSB and CSBDN1 proteins. The central ATPase domain consists of
seven conserved helicase motifs (striped boxes) and is flanked by two putative nuclear localization sequences (gray). (B) Scatter plot showing the
correlation between CSB and CSBDN1 ChIP-seq results: common occupancy sites (black), peaks unique to CSB (red) and peaks unique to CSBDN1
(blue). rpm is reads per million. (C) MSigDB pathways of peaks common to CSB and CSBDN1. doi:10.1371/journal.pgen.1004284.g001 Figure 1. Overview of CSB and CSBDN1 ChIP-seq data. (A) Schematics of CSB and CSBDN1 proteins. The central ATPase domain consists of
seven conserved helicase motifs (striped boxes) and is flanked by two putative nuclear localization sequences (gray). (B) Scatter plot showing the
correlation between CSB and CSBDN1 ChIP-seq results: common occupancy sites (black), peaks unique to CSB (red) and peaks unique to CSBDN1
(blue). rpm is reads per million. (C) MSigDB pathways of peaks common to CSB and CSBDN1. doi:10.1371/journal.pgen.1004284.g001 1.1%, p-value of 4.3e-20 and 0.7% vs 0.3%, p-value of 6.1e-8,
respectively). The occupancy sites unique to CSBDN1 were
slightly over-represented at promoter regions (3% vs 1.1%, p-value
of 2e-06), while the occupancy sites unique to CSB were similar to
the genomic distribution. The genomic occupancy of CSB sites (TSS) and plotted the results as a histogram using a bin size of
500 bp. As shown in Figure S3, CSB displayed only a modest
increase in occupancy around TSSs, while the remodeling-
deficient CSBDN1 derivative displayed greater occupancy around
TSSs. These results are consistent with our functional genomic
distribution analysis, which indicated the CSBDN1 displayed a
more significant enrichment at promoter regions (Figure S2). ATP hydrolysis by CSB is dispensable for chromatin
association during replicative cell growth To validate our ChIP-seq results, we selected seven regions to
analyze by ChIP-qPCR (Figure 2A). chr1-1, chr2-2, chr4-1, and
chr7-1 were four regions that were occupied by both CSB and
CSBDN1. As negative control regions, we examined HES1, chrX-
1, and chr17-1. ChIP-qPCR confirmed that CSB is highly
enriched at chr1-1, chr2-2, chr4-1, and chr7-1 as compared to
HES1, chrX-1, and chr17-1 as well as a ‘‘beads-only’’ control
(Figure 2B). Moreover, like CSB, CSBDN1 was also enriched at
the same four regions (Figure 2B). We next used shRNA-mediated RNA interference to directly
examine the impact of c-Jun on the targeting of CSB to genomic
regions containing a TPA-response element. As shown in Figure 4A,
we were able to significantly reduce c-Jun protein levels through c-
Jun shRNA expression. We used ChIP-qPCR to compare the
recruitment of CSB in cells expressing c-Jun shRNA to cells
expressing a control shRNA. As shown in Figure 4B, there was a
dramatic decrease in CSB enrichment at regions containing the
TGASTCA motif (chr1-1, chr2-2, and chr4-1) or an AP-1-like motif
TGAATCA (chr7-1) in cells expressing c-Jun shRNA as compared
to the control shRNA. On the other hand, there were no significant
changes in the enrichment of CSB at the negative control regions
(HES1, chrX-1, and chr17-1). To demonstrate that c-Jun does,
indeed, occupy these four genomic regions (chr1-1, chr2-2, chr4-1,
and chr7-1), we performed anti-c-Jun ChIP followed by qPCR. As
shown in Figure 4C, c-Jun was enriched at each of these loci. Taken
together, these results indicate that CSB can be targeted to genomic
sites containing a TPA-response element through an association
with a c-Jun-containing AP-1 transcription factor. g
(
g
)
Previously, we found that ATP hydrolysis by CSB is essential for
the recruitment of CSB to UV-induced DNA lesions, a necessary
and early step in the process of transcription-coupled DNA repair
[35]. We, therefore, determined if the ATP hydrolysis activity of
CSB is also important for its targeting to specific genomic regions
in the absence of UV treatment. To accomplish this, we used a
Cockayne syndrome-associated mutant CSB protein, CSBR670W,
which contains a single amino acid substitution at position 670
[36]. This missense mutation, located within the ATPase domain
of CSB, disrupts the ability of CSB to hydrolyze ATP [35]. ATP hydrolysis by CSB is dispensable for chromatin
association during replicative cell growth As
shown in Figure 2C, CSBR670W, like CSB, was targeted to chr1-1,
chr2-2, chr4-1, and chr7-1, but not to the negative control regions
(HES1, chrX-1, and chr17-1) (Figure 2C) [35]. These results
suggest that ATP hydrolysis by CSB is not critical for the
recruitment of CSB to chromatin during replicative cell growth, in
contrast to the recruitment of CSB to DNA lesion-stalled
transcription upon UV irradiation. CSB-PGBD3 is a fusion protein that arises from alternative
splicing between sequence encoding the N-terminal 465 amino
acids of CSB with sequence encoding a piggybac transposase that
lies within the fifth intron of the CSB gene [30,39]. The genomic
sites of CSB-PGBD3 occupancy had been previously determined in
UVSS1KO cells (a CSB and CSB-PGBD3 null cell line) that had
been reconstituted with CSB-PGBD3 [39]. Given that both CSB
and the CSB-PGBD3 fusion protein interact with c-Jun and are
targeted to TPA-response elements, we determined the overlap in
CSB and CSB-PGBD3 occupancy. To accomplish this, we used
HOMER to analyze the published CSB-PGBD3 ChIP-seq data
with the same parameters used to analyze the CSB ChIP-seq data, The sequence-specific transcription factor c-Jun targets
CSB to specific genomic regions Given that CSB does not bind to DNA in a sequence-specific
manner [4,10,11], but is enriched at regions containing the TPA-
response element, this observation suggested that AP-1 transcrip-
tion factors might target CSB to specific genomic loci. To test this
hypothesis, we first determined if CSB could interact with c-Jun,
the most potent transcriptional activator of the Jun protein family
[37,38]. ChIP-western analysis was performed with cells that were
pre-extracted to remove soluble CSB and c-Jun protein before
formaldehyde cross-linking. As shown in Figure 3B, chromatin
immunoprecipitation using an anti-CSB antibody revealed that c-
Jun could be found together with either CSB or CSBDN1 in the
same protein-DNA complex. Co-immunoprecipitation analysis of
unfixed cell lysates, in which the chromatin fraction had been
removed by centrifugation, also revealed an association between
soluble CSB and c-Jun, albeit weaker than that observed in the
chromatin fraction shown by ChIP-western analysis (compare
Figure 3C to 3B). To gain insights into the molecular functions of genes that lie
close to CSB occupancy sites, we searched for overlaps with the
Molecular Signatures Pathways Database (MSigDB) using the
Genomic Regions Enrichment of Annotations Tool (GREAT). The top terms associated with occupancy common to CSB and
CSBDN1 involve the roles of epidermal growth factor receptor
(EGFR) transactivation by G-protein coupled receptors (GPCR),
mechanism of gene regulation by peroxisome proliferators, G-
alpha (12/13) signaling, and NFkB activation (Figure 1C). C-Jun Targets CSB to Regulate Transcription (Table 2) [33,34]. For this analysis, we used the classifications from
normal lung fibroblasts, as CS1AN-sv cells are also a fibroblast cell
line and, therefore, these two lines are most similar [34]. In
agreement with the functional classification described above, CSB
and CSBDN1 displayed significant enrichment at transcribing
promoters (Table 2): 2.7% of the CSB peaks (p-value of 3.2e-58)
and 7.3% of the CSBDN1 peaks (p-value of 4.1e-119) occupied
active and weak promoters. Strikingly, sites of CSB and CSBDN1
occupancy displayed a strong correlation with strong enhancers
(Table 2). Moreover, while regions containing the H3K4me1 (a
mark often associated with enhancers) represented only 4.4% of
fibroblast chromatin, this histone mark was present at ,19% of
the CSB occupancy sites (p-value of 7.9e-1141) and ,26% of
CSBDN1 occupancy sites (p-value of 1.7e-412). Moreover, ,29%
of the top CSB peaks were associated with enhancer features (p-
value of 1.2e-486). Taken together, these results are consistent with
the notion that CSB is involved in the regulation of gene
expression [25–28]. (Figure 3A). This analysis revealed strong enrichment of the TPA
(12-O-tetradecanoylphorbol-13-acetate)-response
element,
TGASTCA (where S denotes a G or C). This motif is most
notable for binding AP-1 (activator protein 1) transcription factors
that consist of either Jun-Jun homodimers or Jun-Fos heterodimers
(Figure 3A) [37,38]. We classified the CSB occupancy sites that
contain TPA-response elements into the same seven functional
categories and found that these sites are slightly over-represented
at transcription termination sites (TTSs) (1.8% vs. 0.9% for a
random genomic sequence, p-value 1.9e-05). CSB occupancy sites
containing TPA-response elements were also under-represented at
39 UTRs (0.7% vs. 1.4%, p-value 9.3e-4) and exons (1.1% vs. 1.9%, p-value 6.1e-04) (Figures S2 and S5). CSB is enriched at sites containing promoter and
enhancer features We next classified the genomic localization of the common and
unique occupancy sites into the seven functional categories
described above (Figure S4). We found that the common CSB
and CSBDN1 occupancy sites were over-represented at promoters
and 59UTRs as compared to the genomic distribution (2% vs To gain further insight into the potential functions of CSB, we
classified CSB and CSBDN1 occupancy sites according to the 15
chromatin states defined by Ernst et al. (2011), which are largely
based on the presence or absence of specific histone marks Table 1. Summary of sequence reads from CSB and CSBDN1 ChIP-seq. ChIP-seq
Total reads
Mapped reads
Unique reads
Total peaks
Unique peaks
Common peaks
CSB
26,707,065
20,511,328
14,569,874
17,779
6,398 (36%)
11,381 (64%)
CSBDN1
63,244,095
43,077,476
34,183,773
3,607
877 (24%)
2,730 (76%)
doi:10.1371/journal.pgen.1004284.t001
PLOS Genetics | www.plosgenetics.org
3
April 2014 | Volume 10 | Issue 4 | e1004284 Table 1. Summary of sequence reads from CSB and CSBDN1 ChIP-seq. April 2014 | Volume 10 | Issue 4 | e1004284 3 C-Jun Targets CSB to Regulate Transcription CSB and CSBDN1 are enriched at genomic regions
containing a TPA-response element To better understand the mechanisms by which CSB is targeted
to specific genomic regions during replicative cell growth, we used
HOMER to determine if binding motifs for sequence-specific
transcription factors were enriched at sites of CSB occupancy April 2014 | Volume 10 | Issue 4 | e1004284 PLOS Genetics | www.plosgenetics.org PLOS Genetics | www.plosgenetics.org 4 C-Jun Targets CSB to Regulate Transcription Table 2. Classification of CSB and CSBDN1 peaks according to chromatin features defined in Ernst et al. (2011). chromatin features
all CSB peaks
top CSB peaks1
all CSBDN1 peaks
chromatin state
enriched chromatin
marks2
% coverage
% coverage
fold3
p-value
% coverage
fold3
p-value
% coverage
fold3
p-value
1. Active Promoter
H3K4me2-3, H3K27Ac,
H3K9Ac
0.6
1.6
2.5
1.9e-30
2.5
4.0
5.1e-26
4.6
7.3
8.4e-87
2. Weak Promoter
H3K4me2-3
0.5
1.1
2.1
7.3e-21
1.8
3.4
4.2e-15
2.6
4.9
5.2e-35
Transcribing promoter (1,2)
H3K4me2-3
1.1
2.7
2.3
3.2e-58
4.3
3.8
1.8e-39
7.3
6.3
4.1e-119
3. Poised Promoter
H3K27me3, H3K4me2
0.1
0.2
1.4
3.1e-02
0.2
1.1
4.8e-1
0.3
2.4
5.5e-3
4. Strong Enhancer
H3K4me1-3, H3K27Ac,
H3K9Ac
0.3
2.2
6.4
1.1e-171
3.2
9.4
2.3e-64
3.1
9.2
2.9e-69
5. Strong Enhancer
H3K4me1, H3K27Ac
1.3
8.8
7.0
2.1e-769
15.7
12.4
8.5e-381
12.4
9.8
3.4e-287
6. Weak Enhancer
H3K4me1, H3K27me2
0.8
2.5
2.9
9.2e-84
3.1
3.6
1.4e-27
3.5
4.2
2.8e-41
7. Weak Enhancer
H3K4me1
1.9
5.6
2.9
3.8e-191
6.9
3.6
6.7e-60
6.0
3.1
8.8e-48
all enhancer (4–7)
H3K4me1
4.4
19
4.4
7.9e-1141
29.2
6.7
1.2e-486
25.5
5.8
1.7e-412
8. Insulator
CTCF
0.9
2.5
2.7
2e-72
1.9
2.1
2.8e-8
2.7
2.9
3.6e-20
9. Transcription Transition
H3K36me3, H4K20me1,
H3K4me1
0.5
1.6
3.0
3.2e-59
1.8
3.3
5.9e-15
1.8
3.4
6.1e-17
10. Transcription Elongation
H3K36me3
4.4
6.2
1.4
8e-28
5.6
1.3
5.4e-4
6.1
1.4
4.5e-7
11. Weak Transcription
none
12.8
12.4
1.0
4.9e-2
10.3
0.8
1.8e-5
9.5
0.7
2.9e-9
12. Repressed
H3K27me3
6.4
11.7
1.8
1.9e-152
10.6
1.7
2.4e-20
10.4
1.6
9.5e-21
13. Heterochrom/lo
none
65.0
42.9
0.7
6.7e-770
33.9
0.5
2.6e-278
33.3
0.5
2.1e-312
14. Repetitive/CNV
medium overall marks
0.1
0.2
1.5
8.4e-3
0.6
4.4
8.3e-8
1.0
6.9
2.1e-18
15. Repetitive/CNV
high overall marks
0.1
0.3
2.5
1.1e-7
0.9
8.8
3.5e-18
1.0
10.4
5.6e-26
1top CSB peaks include only CSB peaks with rpm above 1, and there are 3302 peaks in total. 2partial list, for a detailed description please see Ernst et al. (2011) [34]. April 2014 | Volume 10 | Issue 4 | e1004284 CSB and CSBDN1 are enriched at genomic regions
containing a TPA-response element 3Enrichment: % CSB, top CSB and CSBDN1 peak coverage divided by % genome-wide occurrence. doi:10.1371/journal.pgen.1004284.t002 April 2014 | Volume 10 | Issue 4 | e1004284 April 2014 | Volume 10 | Issue 4 | e1004284 5 C-Jun Targets CSB to Regulate Transcription Figure 2. Validation of ChIP-seq results by ChIP-qPCR. (A) Screen shots of ChIP-seq results displayed from the UCSC genome browser. Shown
are seven different regions with associated transcripts. The y-axis unit is reads per million (rpm). The x-axis represent the genomic positions in base
pairs as follows: HES1_chr3:193,853,385–193,854,474; chrX:73,766,044–73,767,073; chr17:49,769,851–49,771,080; chr1:236,260,208–236,261,267;
chr2:180,324,972–180,325,981; chr4:72,977,907–72,978,936; chr7:2,001,209–2,002,238. The first three regions (HES1, chrX-1, and chr17-1) scored as
negative for CSB and CSBDN1 occupancy and the last four regions (chr1-1, chr2-2, chr4-1, and chr7-1) scored as positive for CSB and CSBDN1
occupancy (ChIPed DNA versus input DNA). The directions of transcription and gene annotation are noted at the bottom. (B) Bar graphs showing CSB
and CSBDN1 ChIP-qPCR results with associated beads-only controls of the seven genomic regions shown in A. (C) Bar graphs showing CSBR670W ChIP-
qPCR results with associated beads-only controls of the seven genomic regions shown in A. The primers used in the qPCR assays are listed in Table
S5. Shown are means +/2 SEM. doi:10.1371/journal.pgen.1004284.g002 igure 2. Validation of ChIP-seq results by ChIP-qPCR. (A) Screen shots of ChIP-seq results displayed from the UC Figure 2. Validation of ChIP-seq results by ChIP-qPCR. (A) Screen shots of ChIP-seq results displayed from the UCSC genome browser. Shown
are seven different regions with associated transcripts. The y-axis unit is reads per million (rpm). The x-axis represent the genomic positions in base
pairs as follows: HES1_chr3:193,853,385–193,854,474; chrX:73,766,044–73,767,073; chr17:49,769,851–49,771,080; chr1:236,260,208–236,261,267;
chr2:180,324,972–180,325,981; chr4:72,977,907–72,978,936; chr7:2,001,209–2,002,238. The first three regions (HES1, chrX-1, and chr17-1) scored as
negative for CSB and CSBDN1 occupancy and the last four regions (chr1-1, chr2-2, chr4-1, and chr7-1) scored as positive for CSB and CSBDN1
occupancy (ChIPed DNA versus input DNA). The directions of transcription and gene annotation are noted at the bottom. (B) Bar graphs showing CSB
and CSBDN1 ChIP-qPCR results with associated beads-only controls of the seven genomic regions shown in A. (C) Bar graphs showing CSBR670W ChIP-
qPCR results with associated beads-only controls of the seven genomic regions shown in A. The primers used in the qPCR assays are listed in Table
S5. Shown are means +/2 SEM. doi:10.1371/journal.pgen.1004284.g002 and we identified 1,590 CSB-PGBD3 ChIP-seq peaks. CSB and CSBDN1 are enriched at genomic regions
containing a TPA-response element Of those
peaks, 165 were common to CSB and CSB-PGBD3 (Tables 3 and
S3). And, of the common peaks, 45% contained a TPA-response
element. coupled with quantitative PCR (RT-qPCR), we compared RNA
expression levels of 10 genes, which displayed significant CSB
occupancy, in CS1AN-sv cells expressing CSB to those in CS1AN-
sv cells harboring an empty vector [8]. For these experiments,
RNA expression was normalized to b-actin transcript levels. Gene ontology analysis using GREAT revealed the top three
‘‘Biological Processes’’ associated with common CSB and CSB-
PGBD3
occupancy
sites
were
tissue
development,
positive
regulation of developmental processes and positive regulation of
catecholamine secretion (Table 4). GREAT was also used to
compare human genes enriched for CSB and CSB-PGBD3 to
genes involved in mouse phenotypes. Of interest, the top three
mouse phenotypes associated with common CSB and CSB-
PGBD3 occupancy were decreased body weight, abnormal body
weight and decreased bone marrow cell number (Table 4). Whether or not deregulation of the genes occupied by both CSB
and CSB-PGBD3 contribute to the clinical features associated
with Cockayne syndrome awaits further studies. ZNFX-NC1 and MCPH1 are two examples of genes that were
positively regulated by CSB (Figure 5A). The ZNFX-NC1 gene
expresses a noncoding RNA that is subsequently processed into
three
snoRNAs
and
is
involved
in
cell
proliferation
and
differentiation [40], and our ChIP-seq results revealed that CSB
binds to the first exon/intron junction of this gene. CSB is also
associated with an intronic region of the microcephalin 1
(MCPH1) gene, which encodes a DNA damage response protein
that may be involved in neurogenesis and the regulation of
cerebral cortex size [41]. RT-qPCR revealed that re-introducing
CSB into CS1AN-sv cells increased the expression of ZNFX-NC1
and MCPH1 about two-fold relative to the vector-only control. Also shown in Figure 5A are examples of eight genes that are
negatively regulated by CSB. ZNF507 and ZNF385B are two zinc
finger proteins likely involved in transcription regulation [42,43]. MSANTD3 is a Myb/SANT-like DNA-binding domain contain- CSB can regulate nearby transcription MAD1L1
(Mitotic Arrest Deficient-Like 1) is a component of the spindle
assembly checkpoint [45]. SACM1L (suppressor of actin muta-
tions 1-like) is a phosphoinositide phosphatase, which degrades
phosphoinositides and plays a key role in signal transduction
events [46]. PRMT5 is a protein arginine methyltransferase and
plays a role in early development and pluripotency [47]. WDR74
(WD Repeat Domain 74) likely plays an essential role in RNA
transcription, stability, and/or processing [48]. DPP9 (dipeptidyl
peptidase 9) regulates signaling pathways that affect cell survival
and proliferation [49]. Among these genes, CSB occupies the
intronic regions of ZNF507, MAD1L1 and ZNF385B, and the
promoter regions of MSANTD3, SACM1L, PRMT5, WDR74,
and DPP9. RT-qPCR analysis revealed that CSB decreased
expression of these genes between 10% and 85%. Altogether, our
results revealed that CSB can both positively and negatively
regulate the expression of genes that lie adjacent to its occupancy
sites. (Tables 6 and S4); among them, 10% and 7%, respectively, were
associated with TPA-response elements. Taken together, these
analyses suggest that at least 25% of the CSB-mediated gene
expression changes might directly result from CSB occupancy. p
g
g
y
p
y
The Bailey et al. (2012) study also examined the transcription
profile of USS1KO cells coexpressing CSB and CSB-PGBD3 as
well as USS1KO cells expressing CSB-PGBD3 alone. When we
compared the CSB occupancy data with those transcription
profiling data (Tables 6 and S4), we found that 22% of the genes
up-regulated and 30% of genes down-regulated by co-expression
of CSB and CSB-PDGB3 were associated with CSB occupancy,
and 19% of the genes up-regulated by CSB-PGBD3 alone and
30% of the genes down-regulated by CSB-PGBD3 alone were
associated with CSB occupancy. These observations are consistent
with the hypothesis that CSB and CSB-PGBD3 may work
together to regulate the transcription of certain genes [29]. Additionally, TPA-response elements were significantly enriched
at CSB-occupied genes that were transcriptionally upregulated by
both CSB and CSB-PGBD3 or by CSB-PGBD3 alone. To identify, on a global level, potential direct transcriptional
targets of CSB, we compared our ChIP-seq data to publicly
available CSB microarray data [28,29]. CSB-dependent tran-
scription profiling had been performed in both hTERT immor-
talized CS1AN fibroblasts (as compared to SV40-transformed
CS1AN cells used in our ChIP-seq study) and SV40 transformed
UVSS1KO fibroblasts. CSB can regulate nearby transcription We next determined the extent to which CSB occupancy can
impact the expression of nearby genes. Using reverse transcription April 2014 | Volume 10 | Issue 4 | e1004284 PLOS Genetics | www.plosgenetics.org 6 C-Jun Targets CSB to Regulate Transcription Figure 3. CSB and CSBDN1 are enriched at sites containing the c-Jun/AP-1 binding motif (TPA-response element). (A) Motif
enrichment at CSB and CSBDN1 occupancy sites. Shown are the fold enrichments with associated p-values. (B) ChIP-western analysis of CSB and
CSBDN1 association with c-Jun on chromatin. (C) Western blot analysis showing c-Jun and CSB co-immunoprecipitation from the soluble (chromatin-
free) fraction of cell lysates. doi:10.1371/journal.pgen.1004284.g003 Figure 3. CSB and CSBDN1 are enriched at sites containing the c-Jun/AP-1 binding motif (TPA-response element). (A) Motif
enrichment at CSB and CSBDN1 occupancy sites. Shown are the fold enrichments with associated p-values. (B) ChIP-western analysis of CSB and
CSBDN1 association with c-Jun on chromatin. (C) Western blot analysis showing c-Jun and CSB co-immunoprecipitation from the soluble (chromatin-
free) fraction of cell lysates. doi:10 1371/journal pgen 1004284 g003 Figure 3. CSB and CSBDN1 are enriched at sites containing the c-Jun/AP-1 binding motif (TPA-response element). (A) Motif
enrichment at CSB and CSBDN1 occupancy sites. Shown are the fold enrichments with associated p-values. (B) ChIP-western analysis of CSB and
CSBDN1 association with c-Jun on chromatin. (C) Western blot analysis showing c-Jun and CSB co-immunoprecipitation from the soluble (chromatin-
free) fraction of cell lysates. doi:10.1371/journal.pgen.1004284.g003 ing protein and is associated with brain tumors [44]. MAD1L1
(Mitotic Arrest Deficient-Like 1) is a component of the spindle
assembly checkpoint [45]. SACM1L (suppressor of actin muta-
tions 1-like) is a phosphoinositide phosphatase, which degrades
phosphoinositides and plays a key role in signal transduction
events [46]. PRMT5 is a protein arginine methyltransferase and
plays a role in early development and pluripotency [47]. WDR74
(WD Repeat Domain 74) likely plays an essential role in RNA
transcription, stability, and/or processing [48]. DPP9 (dipeptidyl
peptidase 9) regulates signaling pathways that affect cell survival
and proliferation [49]. Among these genes, CSB occupies the
intronic regions of ZNF507, MAD1L1 and ZNF385B, and the
promoter regions of MSANTD3, SACM1L, PRMT5, WDR74,
and DPP9. RT-qPCR analysis revealed that CSB decreased
expression of these genes between 10% and 85%. Altogether, our
results revealed that CSB can both positively and negatively
regulate the expression of genes that lie adjacent to its occupancy
sites. ing protein and is associated with brain tumors [44]. CSB has chromatin remodeling-dependent and
-independent activities in transcription regulation p
p
g
To determine whether the remodeling activity of CSB is
required for its function in transcription regulation, we used RT-
qPCR to examine the effect of the remodeling defective CSBDN1
protein on RNA expression [8]. For this analysis, we focused on
genes that were occupied by both CSB and CSBDN1, as revealed
by the ChIP-seq analysis. Western blot analysis using an antibody
that recognizes the C-terminal region of CSB demonstrated that
the levels of CSB or CSBDN1 expression in the stable CS1AN-sv
cell lines used for this analysis were similar (Figure 5B). Additionally, immunofluorescence examination of those cell lines
revealed that the number of cells expressing CSB or CSBDN1
were similar (.95%, data not shown). From this analysis, we In the CS1AN/hTERT cells, Newman et al. (2006) identified
188 CSB up-regulated genes and 205 down-regulated genes. As
shown in Table 5 and S4, 37% of the up-regulated genes and 14%
of the down-regulated genes were occupied by CSB; among these
genes, 9% and 4%, respectively, were associated with TPA-
response elements. In the UVSS1KO study, Bailey et al. (2012)
identified 100 CSB up-regulated genes and 184 down-regulated
genes [29]. As shown in Table 6, 33% of the up-regulated genes
and 34% of the down-regulated genes were occupied by CSB April 2014 | Volume 10 | Issue 4 | e1004284 PLOS Genetics | www.plosgenetics.org 7 C-Jun Targets CSB to Regulate Transcription Figure 4. c-Jun is critical for targeting CSB to genomic regions containing the TPA-response element. (A) Western blot showing a
significant reduction in c-Jun protein levels in cells expressing c-Jun shRNA. (B) Bar graphs showing CSB ChIP-qPCR results obtained from cells
expressing a c-Jun shRNA or a control shRNA. A significant reduction in CSB occupancy occurred at four genomic regions containing the TPA-
response element TGASTCA (chr1-1, chr2-2, and chr4-1) or an AP-1 like motif, TGAATCA (chr7-1), only in cells treated with c-Jun shRNA. The seven
regions analyzed are as shown in Figure 2. (C) Bar graphs showing c-Jun ChIP-qPCR results, demonstrating c-Jun occupancy at the four genomic
regions. Shown are means +/2 SEM. doi:10.1371/journal.pgen.1004284.g004 Figure 4. c-Jun is critical for targeting CSB to genomic regions containing the TPA-response element. (A) Western blot showing a
significant reduction in c-Jun protein levels in cells expressing c-Jun shRNA. (B) Bar graphs showing CSB ChIP-qPCR results obtained from cells
h
l h
f
d
d
f
h Figure 4. CSB has chromatin remodeling-dependent and
-independent activities in transcription regulation GO analysis of common CSB and CSB-PGBD3 occupancy.1
Term Name
Binomial Raw P-Value
Go Biological Process
Tissue development
8.40E-07
Positive regulation of developmental process
1.80E-06
Positive regulation of Catecholamine secretion
4.30E-06
Mouse Phenotype
Decreased body weight
3.40E-08
Abnormal body weight
4.50E-08
Decreased bone marrow cell number
3.30E-07
1Analysis was performed using GREAT (v.2.0.2). doi:10.1371/journal.pgen.1004284.t004 Table 4. GO analysis of common CSB and CSB-PGBD3 occupancy.1
Term Name
Binomial Raw P-Value
Go Biological Process
Tissue development
8.40E-07
Positive regulation of developmental process
1.80E-06
Positive regulation of Catecholamine secretion
4.30E-06
Mouse Phenotype
Decreased body weight
3.40E-08
Abnormal body weight
4.50E-08
Decreased bone marrow cell number
3.30E-07
1Analysis was performed using GREAT (v.2.0.2). doi:10.1371/journal.pgen.1004284.t004 Table 4. GO analysis of common CSB and CSB-PGBD3 occupancy.1 Binomial Raw P-Value 1Analysis was performed using GREAT (v.2.0.2). doi:10.1371/journal.pgen.1004284.t004 analyses suggest that CSB binds to the promoter of ZNFX-NC1
and enables the region that spans amplicons 1–3 to become more
MNase-resistant through nucleosome repositioning or nucleosome
assembly, which promotes transcription up-regulation of ZNFX-
NC1. RNA expression levels in response to CSB expression (Figure 5A). Some of the transcript levels were also decreased by CSBDN1, but
to different degrees (Figure 5A). These results suggest that the
chromatin remodeling activity of CSB is important for transcrip-
tion regulation of some genes (Figure 5A, groups (1) and (2)) and
dispensable for the regulation of others (Figure 5A, group (3)). These results also reveal a chromatin remodeling-independent
activity of CSB in transcription regulation. Of note, CSBDN1,
unlike CSB, did not impact MCPH1 gene expression (Figure 5A). Given that we did not see significant enrichment of CSBDN1 at
this locus (data not shown), we cannot distinguish if the chromatin
remodeling activity or another CSB-related function is important
for up-regulating MCPH1 gene expression (Figure 5A). We next examined the MNase sensitivity of a region of the
PRMT5 promoter near a CSB occupancy site. Remodeling by
CSB is partially required for suppression of PRMT5 expression
(Figure 5A). As depicted in Figure 6B, there are two MNase-
resistant regions that lie on either site of the CSB occupancy site. The nucleosome to the right appears to be better positioned than
the nucleosome to the left. As shown in Figure 6B, cells expressing
either CSB or CSBDN1 had similar sensitivity to MNase at the
region covered by amplicons 1 and 2, but both displayed greater
resistance to MNase than CS1AN cells. CSB has chromatin remodeling-dependent and
-independent activities in transcription regulation The region covered by
amplicons 3–5 appeared to be more sensitive to MNase, suggesting
a relatively more open chromatin structure. Examining MNase
sensitivity patterns at the regions covered by amplicons 6 and 7,
we found that cells expressing CSB showed greater MNase
resistance to the region covered by amplicon 6 than cells not
expressing CSB (CS1AN). Interestingly, cells expressing CSBDN1
demonstrated an intermediate enrichment of amplicon 6 (less than
cells expressing CSB but more than CS1AN cells). On the other
hand, enrichment of amplicon 7 was very similar between cells
expressing CSBDN1 and CS1AN cells, while cells expressing CSB
displayed less MNase resistance. Together, these observations
suggest
that
CSB-expressing
cells
have
a
better-positioned
nucleosome around amplicon 6. Given that CSB suppressed
PRMT5 more efficiently than CSBDN1 (Figure 5A), these results
suggest that a better-positioned nucleosome at the region of the
PRMT5 promoter covered by amplicon 6 may facilitate PRMT5
repression. p
g
g
g
p
(
g
)
To further demonstrate that the chromatin remodeling activity
of CSB is important for regulating ZNFX-NC1 expression, we
performed
micrococcal
nuclease
(MNase)
sensitivity
assays. MNase is a nuclease that preferentially digests naked DNA and
leaves nucleosomal DNA intact. ENCODE data obtained from
the extensively studied K562 and GM12878 cell lines indicates
that there are two MNase-resistant regions adjacent to the CSB
occupancy site in the ZNFX-NC1 gene (Figure 6A) [33]: one
MNase-resistant region is centered at position 47,895,320 on
chromosome 20 and a less resistant region about 150 bp
downstream (Figure 6A). To determine the impact of CSB on
local chromatin structure, we treated formaldehyde cross-linked
cells with limiting amounts of MNase and isolated mononucleo-
somal DNA (,150 bp) (Figure S6). Quantitative PCR using
primer sets that span the ,670 bp region surrounding the CSB
occupancy site were used to compare differences in MNase
sensitivity among cells not expressing CSB (CS1AN), expressing
wild-type CSB, or expressing the remodeling-defective CSBDN1
protein. All PCR reactions were normalized to naked genomic
DNA. ChrX-1 and chr17-1 were used as two negative control
regions, as they are not occupied by CSB. We also examined MNase sensitivity near the site of CSB
occupancy in an intron of the MAD1L1 gene, where several
MNase-resistant regions were predicted based on ENCODE data
[33]. RT-qPCR analysis revealed that the remodeling activity of
CSB was partially required for suppression of MAD1L1 expression
(Figure 5A). CSB has chromatin remodeling-dependent and
-independent activities in transcription regulation c-Jun is critical for targeting CSB to genomic regions containing the TPA-response element. (A) Western blot showing a
significant reduction in c-Jun protein levels in cells expressing c-Jun shRNA. (B) Bar graphs showing CSB ChIP-qPCR results obtained from cells
expressing a c-Jun shRNA or a control shRNA. A significant reduction in CSB occupancy occurred at four genomic regions containing the TPA-
response element TGASTCA (chr1-1, chr2-2, and chr4-1) or an AP-1 like motif, TGAATCA (chr7-1), only in cells treated with c-Jun shRNA. The seven
regions analyzed are as shown in Figure 2. (C) Bar graphs showing c-Jun ChIP-qPCR results, demonstrating c-Jun occupancy at the four genomic
regions. Shown are means +/2 SEM. doi:10.1371/journal.pgen.1004284.g004 ZNFX-NC1 RNA levels similar to that of CS1AN-sv cells
harboring an empty vector. ChIP-qPCR confirmed that both
CSB and CSBDN1 were recruited to the ZNFX-NC1 locus
(Figure 5C). The other genes that we examined showed decreased found that expression of the ZNFX-NC1 gene, which was
enhanced by CSB, was not enhanced by CSBDN1 (Figure 5A);
the presence of CSB resulted in a greater than two-fold increase in
ZNFX-NC1 RNA levels, while cells expressing CSBDN1 had found that expression of the ZNFX-NC1 gene, which was
enhanced by CSB, was not enhanced by CSBDN1 (Figure 5A);
the presence of CSB resulted in a greater than two-fold increase in
ZNFX-NC1 RNA levels, while cells expressing CSBDN1 had Table 3. Comparison of CSB and CSBDN1 occupancy with CSB-PGBD3#. ChIP-seq
No. of common
occupancy sites with
CSB-PGBD3#
Percent common
occupancy of CSB or
CSBDN1 with CSB-PGBD3
Percent common
occupancy of CSB-PGBD3
with CSB or CSBDN1$
No. of c-Jun/AP-1
motifs in common
occupancy
p-values1,{,`
CSB
165
0.9%
10.4%
74 (45%)
3.5e-101 4.6e-38{
CSBDN1
81
2.2%
5.1%
42 (52%)
2.1e-81 6.7e-19`
#CSB-PGBD3 ChIP-seq data are from Gray et al. (2012) [39]. $Total number of CSB-PGBD3 occupancy sites is 1590. 1hypergeometric p-value against CSB-PGBGD3 peaks. {hypergeometric p-value against CSB peaks. `hypergeometric p-value against CSBDN1 peaks. doi:10.1371/journal.pgen.1004284.t003
PLOS Genetics | www.plosgenetics.org
8
April 2014 | Volume 10 | Issue 4 | e1004284 Table 3. Comparison of CSB and CSBDN1 occupancy with CSB-PGBD3#. April 2014 | Volume 10 | Issue 4 | e1004284 April 2014 | Volume 10 | Issue 4 | e1004284 C-Jun Targets CSB to Regulate Transcription Table 4. CSB has chromatin remodeling-dependent and
-independent activities in transcription regulation As shown in Figure 6C, cells expressing CSB
demonstrated a similar MNase resistance to cells expressing
CSBDN1 at the region covered by amplicons 2, 3, and 6, but
higher than CS1AN cells. Additionally, CSB-expressing cells
displayed greater MNase sensitivity at the region covered by
amplicons 4 and 5 than CSBDN1 or CS1AN cells, which were
similar to each other. These results suggest that increased
nucleosome occupancy at the region of the MAD1L1 gene
covered by amplicons 4–5 may facilitate MAD1L1 repression. As shown in Figure 6A, cells expressing CSB or CSBDN1, as
well as cells harboring an empty vector, had similar levels of
amplicon enrichment from chr17-1 and chrX-1, indicating similar
MNase sensitivities. However, cells expressing CSB demonstrated
a greater enrichment of amplicons 1, 2, and 3 than cells expressing
CSBDN1 or harboring an empty vector (CS1AN). These results
indicate that this region is more protected from MNase digestion
in CSB expressing cells. On the other hand, enrichment of
amplicons 4 and 5 was very similar between CSB and CS1AN,
while CSBDN1 displayed slightly less MNase resistance. Given
that chromatin remodeling by CSB is required for transcription
up-regulation of ZNFX-NC1 (Figure 5A), our MNase-qPCR April 2014 | Volume 10 | Issue 4 | e1004284 PLOS Genetics | www.plosgenetics.org 9 C-Jun Targets CSB to Regulate Transcription Figure 5. CSB can influence nearby gene expression and has both remodeling-dependent as well as independent functions. (A) Gene
expression was assayed by RT-qPCR using CS1AN-sv cells expressing CSB, CSBDN1, or harboring an empty vector, to determine the effect of CSB and
CSBDN1 on the expression of CSB-occupied genes. Expression levels were normalized to b-actin (ACTB) and are presented as fold change over the
vector only control. Shown are means +/2 SEM. All data are averaged from six biological replicates, and each biological replicate consisted of three
technical replicates. The expression of ten genes was analyzed (see text for the relative positions of CSB occupancy at these genes). The primers used
in RT-qPCR assays are listed in Table S6. CSBDN1 could not enhance expression of the ZNFX1-NC1 or MCPH1 genes, unlike CSB (1), while CSBDN1
could partially substitute for CSB function at the MAD1L1, SACM1L, PRMT5 and ZNF385B genes (2). CSBDN1 could fully substitute for CSB function at
the WDR74 and DDP9 genes (3). CSBDN1 was not examined for ZNF507 or MSANTD3 (*). CSB has chromatin remodeling-dependent and
-independent activities in transcription regulation (B) Western blot analysis demonstrating that the levels of
CSB and CSBDN1 expression in the stable cell lines that were used for these assays were similar. (C) ChIP-qPCR reveals that both CSB and CSBDN1
were recruited to ZNFX1-NC1 (chr20-1) locus. HES1 was used as a negative control region in this assay. doi:10.1371/journal.pgen.1004284.g005 Figure 5. CSB can influence nearby gene expression and has both remodeling-dependent as well as independent functions. (A) Gene
expression was assayed by RT-qPCR using CS1AN-sv cells expressing CSB, CSBDN1, or harboring an empty vector, to determine the effect of CSB and
CSBDN1 on the expression of CSB-occupied genes. Expression levels were normalized to b-actin (ACTB) and are presented as fold change over the
vector only control. Shown are means +/2 SEM. All data are averaged from six biological replicates, and each biological replicate consisted of three
technical replicates. The expression of ten genes was analyzed (see text for the relative positions of CSB occupancy at these genes). The primers used
in RT-qPCR assays are listed in Table S6. CSBDN1 could not enhance expression of the ZNFX1-NC1 or MCPH1 genes, unlike CSB (1), while CSBDN1
could partially substitute for CSB function at the MAD1L1, SACM1L, PRMT5 and ZNF385B genes (2). CSBDN1 could fully substitute for CSB function at
the WDR74 and DDP9 genes (3). CSBDN1 was not examined for ZNF507 or MSANTD3 (*). (B) Western blot analysis demonstrating that the levels of
CSB and CSBDN1 expression in the stable cell lines that were used for these assays were similar. (C) ChIP-qPCR reveals that both CSB and CSBDN1
were recruited to ZNFX1-NC1 (chr20-1) locus. HES1 was used as a negative control region in this assay. doi:10.1371/journal.pgen.1004284.g005 Table 5. Correlation of CSB ChIP-seq data with CSB transcription profiling data of Newman et al. (2006). Number of
genes1
Number of genes associated
with CSB occupancy
Number of CSB- binding sites
associated with CSB-responsive
genes2
Number of c-Jun/AP-1
motifs3
Up-regulated by CSB
188
69 (37%)
196 (1.1%) (p-value: ,1.e-300)
18 (9%) (p-value: 0.08)
Down-regulated by CSB
205
28 (14%)
45 (0.25%) (p-value: 1)
2 (4%) (p-value: 0.17)
1CSB transcription profiling data are from Newman et al. (2006) [28]. 2p-values were calculated using a z-score after randomly generating 17,779 peaks against the genome. 3Hypergeometric p-values were calculated against the total number of CSB ChIP-seq peaks (this study). doi:10.1371/journal.pgen.1004284.t005 ble 5. CSB has chromatin remodeling-dependent and
-independent activities in transcription regulation Correlation of CSB ChIP-seq data with CSB transcription profiling data of Newman et al. (2006). April 2014 | Volume 10 | Issue 4 | e1004284 10 C-Jun Targets CSB to Regulate Transcription Table 6. Correlation of CSB ChIP-seq data with CSB and CSB-PGBD3 transcription profiling data of Bailey et al. (2012). Table 6. Correlation of CSB ChIP-seq data with CSB and CSB-PGBD3 transcription profiling data of Bailey et al. (2012). q
p
p
g
y
(
)
Number of
genes1
Number of genes
associated with CSB
occupancy
Number of CSB- binding
sites associated with CSB-
responsive genes2
Number of c-Jun/AP-1
motifs3
Up-regulated by CSB
100
33 (33%)
73 (0.4%) p-value: 0.67
7 (10%) p-value: 0.13
Down-regulated by CSB
184
63 (34%)
182 (1%) p-value: 0.99
12 (7%) p-value: 0.09
Up-regulated by CSB-PGBD3
248
48 (19%)
81 (0.5%) p-value: 0.27
12 (15%) p-value:0.014
Down-regulated by CSB-PGBD3
273
81 (30%)
307 (1.7%) p-value: 0.99
26 (8%) p-value: 0.07
Up-regulated by CSB+CSB-PGBD3 913
201 (22%)
608 (3.4%) p-value: 1
62 (10%) p-value: 0.006
Down-regulated by CSB+CSB-
PGBD3
329
98 (30%)
297 (1.7%) p-value: 1
19 (6%) p-value: 0.17
1Transcription profiling data are from Bailey et al. (2012) [29]. 2p-values were calculated using a z-score after randomly generating 17,779 peaks against the genome. 3Hypergeometric p-values were calculated against the total number of CSB ChIP-seq peaks (this study). doi:10.1371/journal.pgen.1004284.t006 p
p
g
y
2p-values were calculated using a z-score after randomly generating 17,779 peaks against the genome. 3Hypergeometric p-values were calculated against the total number of CSB ChIP-seq peaks (this study). doi:10.1371/journal.pgen.1004284.t006 Figure 6. CSB can alter the MNase sensitivity of nearby nucleosomes. (A–C) first panels from top to bottom: screen shots from the UCSC
genome browser (GRCh37/hg19 Assembly) showing the RefSeq gene and direction of transcription (arrow head), the position of CSB occupancy (CSB
ChIP), the position of CSBDN1 occupancy (CSBDN1 ChIP), ENCODE MNase-seq data obtained from K562 cells (MNase), and the amplicons used in the
MNase-qPCR assays (Amplicon). The chromosome coordinates shown are (A) Chr20:47,894,930–47,895,599 (B) Chr14:23,398,657–23,399,208 and (C)
Chr7:2,001,747–2,002,144. Second panels in A–C are bar graphs showing results from MNase-qPCR assays. The primers used in the MNase-qPCR
assays are listed in Table S7. Shown are means +/2 SEM. doi:10.1371/journal.pgen.1004284.g006 Figure 6. CSB can alter the MNase sensitivity of nearby nucleosomes. 1Transcription profiling data are from Bailey et al. (2012) [29].
2p-values were calculated using a z-score after randomly generating 17,779 peaks against the genome.
3Hypergeometric p-values were calculated against the total number of CSB ChIP-seq peaks (this study).
doi:10.1371/journal.pgen.1004284.t006 C-Jun Targets CSB to Regulate Transcription are not associated with known CSB-responsive genes (Tables 5–6). This could arise from the different cell lines used in these studies
and/or the different immortalization methods used to obtain these
cell lines. It is also possible that some CSB-responsive genes were
not covered by the microarrays used for the transcription profiling
studies and, therefore, complementary approaches such as RNA-
seq might offer additional insights, such as the influence of CSB on
non-coding RNA expression. Furthermore, our analysis indicates a
strong correlation between sites of CSB occupancy and chromatin
regions that contain epigenetic signatures of enhancers (Table 2),
and many of the CSB peaks (41%, p-value of 6.7e-1536) have
DNase I hypersensitive sites lying within 100 bp, as judged by the
digital DNase I hypersensitivity clusters in 125 cell lines [33]. These observations suggest that some of the intergenic CSB
occupancy sites could function as enhancer elements that might lie
at a great distance from their target genes. are not associated with known CSB-responsive genes (Tables 5–6). This could arise from the different cell lines used in these studies
and/or the different immortalization methods used to obtain these
cell lines. It is also possible that some CSB-responsive genes were
not covered by the microarrays used for the transcription profiling
studies and, therefore, complementary approaches such as RNA-
seq might offer additional insights, such as the influence of CSB on
non-coding RNA expression. Furthermore, our analysis indicates a
strong correlation between sites of CSB occupancy and chromatin
regions that contain epigenetic signatures of enhancers (Table 2),
and many of the CSB peaks (41%, p-value of 6.7e-1536) have
DNase I hypersensitive sites lying within 100 bp, as judged by the
digital DNase I hypersensitivity clusters in 125 cell lines [33]. These observations suggest that some of the intergenic CSB
occupancy sites could function as enhancer elements that might lie
at a great distance from their target genes. Lastly, we examined MNase sensitivity at the promoter of the
WDR74 gene, where remodeling by CSB was dispensable for
CSB-dependent suppression of WDR74 expression. Cells express-
ing CSB or CSBDN1 displayed very similar patterns of MNase
sensitivity, agreeing with the results of our RT-qPCR analysis
(Figure 5A), which revealed that CSB and CSBDN1 had a similar
effect on WDR74 expression (Figure S7). Discussion The composition of the dimeric AP-1 transcription factors that
bind to TPA-response elements varies; for instance, there are three
Jun and four Fos family members, and some of these members
have variants that result from alternative splicing [37,38]. Future
experiments will unveil the full extent to which different AP-1
complexes can target CSB and how this targeting might be
modulated in different cellular contexts. AP-1 participates in a
number of fundamental cellular processes, including cell prolifer-
ation and cell death. It is tempting to speculate that loss of CSB
activity might, to some extent, compromise AP-1-mediated gene
regulation, which in turn might contribute to the underlying
mechanisms of Cockayne syndrome. Our results reveal that the mechanism that targets CSB to
chromatin for transcription regulation is distinct from the
mechanism that targets CSB during transcription-coupled DNA
repair [35]. The association of CSB with specific genomic loci
during replicative cell growth does not rely upon ATP hydrolysis
by CSB (Figure 2C); however, ATP hydrolysis by CSB is essential
for its targeting to sites of UV-induced DNA damage [35]. Therefore, stable chromatin association during transcription-
coupled DNA repair appears to be an active process while stable
chromatin association during transcription regulation appears to
be a passive process. Current evidence suggests that during
transcription-coupled DNA repair, ATP hydrolysis by CSB
induces a conformational change that exposes a chromatin
interaction surface, which is normally occluded by the N-
terminal region of CSB [35]. Given that the association of CSB
with chromatin in the absence of UV-induced DNA lesions
does not rely upon ATP hydrolysis, this would suggest that
the residues that mediate the c-Jun association are normally
exposed. Approximately 15% of the total CSB occupancy sites contain
TPA-response elements. The mechanism that underlies the
targeting of CSB to regions of the genome that do not contain
TPA-response elements is not yet clear, but it is likely that CSB is
delivered to or stabilized at these regions through associations
with other DNA-binding proteins. A recent study examining the
targeting of Isw2 in Saccharomyces cerevisiae revealed that this
remodeler is primarily targeted to specific loci by sequence-
specific transcription factors; however, more than half of the
transcription factor-dependent occupancy sites did not contain a
cognate binding motif [50]. Discussion Chromatin conformation capture
suggested that DNA looping between regions that contain a
transcription factor binding site with regions that do not is an
integral component of the Iswi2 targeting mechanism [50]. Accordingly, DNA looping may also play an important role in
the targeting of the CSB remodeler to sites that do not contain a
TPA-response
element. Interestingly,
AP-1,
in
conjunction
with NFkB, was found to mediate DNA looping to regulate
gene expression in macrophages [51]. Future studies examining
CSB-containing protein complexes and higher-order chromatin
structure
will
offer
insights
into
other
CSB
targeting
mechanisms. Recently, Gray et al. showed that the N-terminal region of
CSB mediates the interaction between c-Jun and CSB-PGBD3
[39]. Based upon this observation and the knowledge that the
CSB-PGBD3 fusion protein contains the N-terminal 465 amino
acids of CSB, it is likely that the N-terminal 465 amino acids also
mediates the association of full-length CSB with c-Jun. However,
an interaction between full-length CSB and c-Jun was not
detected in that study, although a robust interaction between
CSB-PGBD3 and c-Jun was observed [39]. In agreement with
that observation, we observed only modest association between
endogenous CSB and c-Jun in chromatin-free cell lysates
(Figure 3C). However, a greater degree of association was
observed
when
we
specifically
examined
chromatin-bound
proteins (Figure 3B). These observations suggest that the CSB-
c-Jun association may be preferentially established or stabilized
on chromatin (Figure 7). Our ChIP-seq data revealed that 36% of the CSB peaks were
unique to CSB and 24% of the CSBDN1 peaks were unique to
CSBDN1 (Figure 1B and Table 1). We do not yet know the reason
underlying this difference. It is possible that some of the occupancy
sites unique to CSBDN1 might represent the initial sites of CSB
binding to chromatin and that CSB would subsequently translo-
cate away from these sites during chromatin remodeling, which in
turn might contribute to some of the unique CSB peaks. Alternatively, but not mutually exclusive, some of the unique
CSB peaks might represent targeting that relies upon functions
related to the N1 region, which is deleted in the CSBDN1 protein,
such as mediating protein-protein interactions. Of interest,
CSBDN1 is more significantly enriched at promoters and 59
UTR regions than CSB (Figures S2, S3). CSB and CSB-PGBD3 both interact with c-Jun, and 45% of
peaks common to CSB and CSB-PGBD3 contain a TPA-
responsive element (Table 3) [39]. C-Jun Targets CSB to Regulate Transcription Interestingly, the
nucleosome structure at the promoter of the WDR74 gene
appeared to be relatively more open, as the overall MNase-qPCR
signals were lower than those obtained from the ZNFX-NC1,
PRMT5 and MAD1L1 genes (Figure 6). A more open nucleosome
structure could account for, at least in part, our observation that
suppression of WDR74 expression by CSB does not rely upon
remodeling activity of CSB. CSB has chromatin remodeling-dependent and
-independent activities in transcription regulation (A–C) first panels from top to bottom: screen shots from the UCSC
genome browser (GRCh37/hg19 Assembly) showing the RefSeq gene and direction of transcription (arrow head), the position of CSB occupancy (CSB
ChIP), the position of CSBDN1 occupancy (CSBDN1 ChIP), ENCODE MNase-seq data obtained from K562 cells (MNase), and the amplicons used in the
MNase-qPCR assays (Amplicon). The chromosome coordinates shown are (A) Chr20:47,894,930–47,895,599 (B) Chr14:23,398,657–23,399,208 and (C)
Chr7:2,001,747–2,002,144. Second panels in A–C are bar graphs showing results from MNase-qPCR assays. The primers used in the MNase-qPCR
assays are listed in Table S7. Shown are means +/2 SEM. doi:10.1371/journal.pgen.1004284.g006 April 2014 | Volume 10 | Issue 4 | e1004284 PLOS Genetics | www.plosgenetics.org 11 11 PLOS Genetics | www.plosgenetics.org Figure 7. Model for the targeting of CSB to chromatin for transcription regulation. The sequence-specific transcription factor c-Jun
interacts with CSB (directly or indirectly) and either delivers CSB to chromatin or is pre-bound and stabilizes the association of CSB with chromatin at
regions containing the TPA-response element. This targeting mechanism is independent of ATP hydrolysis by CSB and, therefore, distinct from CSB
targeting to chromatin in response to UV irradiation, which requires ATP hydrolysis. Once chromatin associated, CSB can either activate or repress the
expression of nearby genes. The function of CSB in regulating gene expression relies upon both remodeling-dependent and remodeling-
independent activities of CSB. doi:10.1371/journal.pgen.1004284.g007 By examining the effect of CSB on several genes that lie close
to CSB occupancy sites, we provide evidence that CSB can
directly influence local gene expression mediated by RNA
polymerase II. CSB is a member of the SWI2/SNF2 ATP-
dependent chromatin remodeling protein family and displays
ATP-dependent chromatin remodeling activity in vitro; therefore,
CSB likely repositions nucleosomes [8,13] and/or other protein
factors [17] to regulate transcription. By mapping nucleosome
positions at the ZNFX-NC1, PRMT5 and MAD1L1 genes with
MNase, we found that regions adjacent to the CSB occupancy
sites are more resistant to MNase digestion in cells expressing
CSB than in cells that do not express CSB or express the
remodeling-defective CSBDN1 protein. These results indicate
that CSB alters chromatin structure in an ATP-dependent
manner to regulate transcription (Figures 6–7). Furthermore,
CSB appears to make the border of nucleosome-free regions
more pronounced (Figure 6A amplicon 2–3 and Figure 6B
amplicon 6), resembling the function of yeast Iswi2 in regulating
the
length
of
nucleosome-free
regions
to
prevent
cryptic
transcription and regulate gene expression [52]. By creating a
better-positioned nucleosome (Figure 6C, amplicon 4–5), CSB
could also support either transcription activation or repression by
preventing the binding of transcriptional repressors or activators,
respectively, to the DNA occupied by the nucleosome [53]. Interestingly, in collaboration with NAP1-like histone chaper-
ones, CSB has been shown to efficiently move histone octamers to
the center of a DNA fragment in vitro [8]. It will be of great
interest to investigate the function of NAP1-like chaperones in
CSB-mediated transcription regulation. Additionally, the com-
plete rescue of gene expression by the CSBDN1 protein at certain
genes (e.g. Discussion These observations are
consistent with results obtained from transcription profiling
studies, in which it was observed that CSB and CSB-PGBD3
can co-regulate the expression of certain genes [29]. These results
further suggest that AP-1 transcription factors might play a crucial
role in modulating this co-regulation. Additionally, these results
are also consistent with the notion that CSB regulates many genes
independently of CSB-PGBD3 [29]. From comparisons between ChIP-seq and transcription profil-
ing data, it can be seen that the majority of CSB occupancy sites April 2014 | Volume 10 | Issue 4 | e1004284 12 PLOS Genetics | www.plosgenetics.org C-Jun Targets CSB to Regulate Transcription By examining the effect of CSB on several genes that lie close
CSB
i
id
id
h
CSB
Interestingly, in collaboration with NAP1-like histone chaper-
CSB h
b
h
ffi i
l
hi
Figure 7. Model for the targeting of CSB to chromatin for transcription regulation. The sequence-specific transcription factor c-Jun
interacts with CSB (directly or indirectly) and either delivers CSB to chromatin or is pre-bound and stabilizes the association of CSB with chromatin at
regions containing the TPA-response element. This targeting mechanism is independent of ATP hydrolysis by CSB and, therefore, distinct from CSB
targeting to chromatin in response to UV irradiation, which requires ATP hydrolysis. Once chromatin associated, CSB can either activate or repress the
expression of nearby genes. The function of CSB in regulating gene expression relies upon both remodeling-dependent and remodeling-
independent activities of CSB. doi:10.1371/journal.pgen.1004284.g007
C-Jun Targets CSB to Regulate Transcription WDR74) suggests additional CSB functions in
transcription regulation; such functions could include protein
recruitment
through
remodeling-independent
mechanisms
[12,54] or protein eviction [55], which may not rely upon the
N1 region. During replicative cell growth, approximately 10% of CSB
associates with chromatin, and this likely represents the CSB
population that participates in normal transcription regulation. However, in the presence of UV-induced DNA damage (.25 J/
m2), approximately 90% of the CSB population can become stably
associated with chromatin [35]. A fraction of these chromatin-
associated CSB molecules would be stabilized at sites of DNA
lesion-stalled transcription to participate in DNA repair. In
addition, some of these CSB molecules would also be expected
to localize to new transcriptional targets, as CSB has been
implicated in UV-induced transcription regulation [17]. Addition-
al ChIP-seq analysis of CSB in cells challenged with UV
irradiation will reveal if the fraction of CSB that is used during April 2014 | Volume 10 | Issue 4 | e1004284 13 PLOS Genetics | www.plosgenetics.org C-Jun Targets CSB to Regulate Transcription normal transcription regulation is redistributed in response to UV
irradiation, either for DNA repair or UV-induced transcription
regulation. ChIP and ChIP-western analysis y
To increase ChIP efficiency we removed soluble CSB before
cross-linking DNA and proteins [8,35,58]. Cells were collected in
Buffer B (150 mM NaCl, 0.5 mM MgCl2, 20 mM HEPES
pH 7.8, 10% Glycerol, 0.5% Triton X-100) and soluble CSB
was separated from chromatin by centrifugation at 15,000 RPM
for 5 min at 4uC. The resulting pellets were resuspended in Buffer
B and fixed with 1% formaldehyde for 10 min at room
temperature. Cross-linked cells were sonicated at 40% amplitude
(30 sec on, 90 sec off, for 24 min total) using the Branson 101-135-
126 Sonifier. Chromatin IP (ChIP) was performed using a
monoclonal anti-CSB antibody (1B1) that recognizes the N-
terminal 507 amino acids of CSB [35,39] or an anti-c-Jun
antibody (Santa Cruz, sc-1694). ChIP samples were reverse cross-
linked in SDS sample buffer for subsequent western blot analyses
[20]. Antibodies used for western blot analysis are rabbit anti-CSB
antibodies (kindly provided by Dr. Alan Weiner, U. Washington)
[35], c-Jun (Santa Cruz, sc-1694) and GAPDH (Millipore,
MAB374). g
Taken together, the results of this study reveal that the CSB
remodeler binds to specific regions of the genome to regulate
chromatin structure and RNA polymerase II-mediated gene
expression. shRNA knockdown Mission shRNA targeting c-Jun (TRCN0000010366, Sigma)
was used to decrease c-Jun protein levels. A non-targeting shRNA
(SHC002, Sigma) was used as a negative control. Virus was
produced by cotransfecting a 10 cm plate of ,90% confluent
293T cells with third generation lentivirus packaging plasmids
(pMGLg/pRRE, pRSV-REV, and pMD2.G/VSV). A total of
20 mg of plasmid was transfected, with the individual plasmids at
an equal molar ratio. The culture medium was changed 24 hours
post-transfection, and virus-containing medium was collected
24 hours later. Medium from one plate of virus-producing cells
was distributed to six 10 cm dishes of target cells: CS1AN-sv/
CSB. The confluence of the target cells at the time of infection
was approximately 20%. Infected cells were harvested 36–
48 hours post-infection for RNA preparation and western blot
analysis. To classify a peak as unique or common, we determined the
intensities (rpm) of the CSB and CSBDN1 signals within a 200 bp
region around a peak center. If the difference between signal
intensities was 4-fold or greater and the p-value for that difference
was #0.0001, the peak was classified as unique. The remaining
signals were classified as common. CSB and CSBDN1 peaks were
classified as follows: (1) promoter (from 21 kb to the transcription
start site), (2) TTS (from the transcription termination site to +
1 kb), (3) 59 UTR, (4) 39 UTR, (5) exon, (6) intron, and (7)
intergenic (the remainder). The source of gene annotation was
UCSC RefGene. The CEAS package [48] computes p-values
using one-sided binomial test. To compute p-values for compar-
isons between CSB and CSBDN1, we considered a null model in
which both classes of peaks form a single population. m and n are
the total peak numbers of those classes, and m9 and n9 are the
number of peaks in a specific annotation category, and a
combined frequency f equal to (m9+n9)/(m+n). p-values for
comparisons between CSB and CSBDN1 are equal to the product
of the two p-values from the one-sided binomial test for n, n9 and f,
as well as for m, m9 and f. Cell culture CS1AN-sv cells were maintained in DMEM-F12 with 10%
FBS. The CS1AN primary cells have mutations in both CSB
alleles, but only one of these alleles was retained after SV40
immortalization [19]; the resulting CS1AN cell line is, therefore,
hemizygous for CSB. The retained allele contains an A to T
transversion at position 1088, which introduces a premature stop
codon at amino acid 337. Accordingly, CSB-PGBD3 is predicted
to be absent from the CS1AN-sv cell line, and our anti-CSB
immunoprecipitation experiments agree with this prediction
(Figure S1). ChIP-seq and data analysis 10 ng of ChIPed DNA was used to prepare libraries for deep
sequencing using the multiplexed ChIP-Seq sample preparation
protocol described on the website of the Next-Generation
Sequencing Core, Perelman School of Medicine, University of
Pennsylvania (http://ngsc.med.upenn.edu/). The Next-Genera-
tion Sequencing Core at the University of Pennsylvania performed
DNA sequencing using Illumina hiSeq2000 sequencers for single-
end sequencing with a read length of 50 bps. The resulting
sequencing reads were mapped to the human genome (HG19
assembly) using Bowtie version 0.12.7. Peaks were identified using
HOMER version 4.1 (Hypergeometric Optimization of Motif
EnRichment) with a default option (FDR = 0.001 and Poisson p-
value cutoff = 0.0001) on ChIPed samples against matching input
DNA samples. Raw and processed files (GSE50171) have been
deposited at the Gene Expression Omnibus (GEO) repository. (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?token = vfmfry
agmygcony&acc = GSE50171). Stable cell lines expressing CSB were generated by infecting
CS1AN-sv cells with CSB-expressing lentivirus (pLenti-PGK-Neo,
Addgene) [8]. Stable cell lines expressing CSB or harboring the
empty vector were selected with 600 mg/ml G418. CSBDN1 was
expressed from the pSVL vector [8]. CS1AN-sv cells stably
expressing CSBDN1 were generated by cotransfection with
pLenti-PGK-neo. After selection with 600 mg/ml G418, single
colonies were cloned [8]. CSBR670W was expressed from MSCV-
Puro. The stable cell line expressing CSBR670W was generated by
transfection and selecting with 250 ng/ml puromycin [35]. These observations are consistent with the hypothesis
that Cockayne syndrome might be, at least in part, a disease of
transcription deregulation [28,29,56]. Moreover, the results of
this study open up new avenues to explore the mechanisms
that might contribute to the diverse features of Cockayne
syndrome. (TIFF) Figure S6
Preparation of mononucleosomal DNA for MNase
sensitivity assays. (A) After limited MNase digestion of formaldehyde
cross-linked nuclei, the cross-links were reversed, and the DNA was
purified and resolved in an agarose gel. (B) Mononucleosomal DNA
purified from (A) and used in the qPCR assays. (PDF) Figure S7
MNase-qPCR analysis of the WDR74 promoter
region. Upper panel from top to bottom: screen shots from the
UCSC genome browser (GRCh37/hg19 Assembly) showing the
RefSeq gene and direction of transcription (arrow head), the
position of CSB occupancy (CSB ChIP), the position of CSBDN1
occupancy (CSBDN1 ChIP), ENCODE MNase-seq data obtained
from K562 cells (MNase), and the amplicons used in the MNase-
qPCR assays (Amplicon). The chromosome coordinates shown are
chr11:62,608,860-62,609,234. Lower panel contains bar graphs
showing results from the MNase-qPCR assays. The primers used
in the MNase-qPCR assays are listed in Table S7. Shown are
means +/2 SEM. (TIFF) Co-immunoprecipitation assays CS1AN cells expressing CSB were lysed in Buffer B60 (20 mM
HEPES (pH 7.9), 60 mM NaCl, 0.5 mM MgCl2, 1 mM DTT,
0.5% triton X-100 and 20% glycerol) with protease inhibitors
(0.5 mM PMSF, 10 mM E64 and 3 mM pepstatin A) and
centrifuged at 15,000 rpm for 5 min at 4uC. The supernatant
was used for co-immunoprecipitation assays. CSB-containing
complexes
were
recovered using
the
monoclonal
anti-CSB
antibody 1B1 (1 hour at 4uC) and protein G agarose beads
(30 min at 4uC). The resulting immunocomplexes were washed
four times in buffer B60, and the immunoprecipitated proteins
were eluted with Laemmli buffer. Beads-only control immuno-
precipitations were conducted in parallel. CS1AN cells expressing CSB were lysed in Buffer B60 (20 mM
HEPES (pH 7.9), 60 mM NaCl, 0.5 mM MgCl2, 1 mM DTT,
0.5% triton X-100 and 20% glycerol) with protease inhibitors
(0.5 mM PMSF, 10 mM E64 and 3 mM pepstatin A) and
centrifuged at 15,000 rpm for 5 min at 4uC. The supernatant
was used for co-immunoprecipitation assays. CSB-containing
complexes
were
recovered using
the
monoclonal
anti-CSB
antibody 1B1 (1 hour at 4uC) and protein G agarose beads
(30 min at 4uC). The resulting immunocomplexes were washed
four times in buffer B60, and the immunoprecipitated proteins
were eluted with Laemmli buffer. Beads-only control immuno-
precipitations were conducted in parallel. Figure S2
Genomic distributions of CSB and CSBDN1. The
genome was divided into seven categories defined by the UCSC
RefSeq gene annotation. The genomic distributions of CSB and
CSBDN1 were determined using the CEAS package and are
presented as pie charts [32]. The table includes p-values for these
distributions. (TIFF) Comparison of the CSB ChIP-seq data to published CSB-
PGBD3 ChIP-seq data Comparison of the CSB ChIP-seq data to published CSB-
PGBD3 ChIP-seq data To compare the genomic occupancy of CSB to CSB-PGBD3,
we downloaded the ChIP-seq data for CSB-PGBD3 and the
matching input from the GEO repository (GSE37919) [39]. The
CSB-PGBD3 peaks were called against input using HOMER, and
we identified 1,590 peaks. Using binary peak calling (+/2100 bp),
we identified 165 peaks as common to CSB and CSB-PGBD3,
which represented 1% total CSB and 10% total CSB-PGBD3
peaks (Tables 3 and S3). The hypergeometric p-value was
calculated for the AP-1 motif against total CSB as well as CSB-
PGBD3 peaks (Table 3). The ontology of the nearby genes was
obtained using GREAT (Table 4). Supporting Information Figure S1
CSB-PGBD3 is not expressed in CS1AN-sv cells
reconstituted with CSB. Whole-cell lysates from 293T cells,
CS1AN-sv cells, and CS1AN-sv cells reconstituted with CSB were
subjected to immunoprecipitation using the monoclonal anti-CSB
N-terminal antibody 1B1. The immunoprecipitated material,
along with the input lysates, was resolved in a 7% Tris-Acetate
gel (NuPAGE) and the western blot was probed with the
polyclonal
anti-CSB
N-terminal
antibody. The
monoclonal
antibody immunoprecipitated both CSB and CSB-PGBD3 from
293T cells, but this antibody only immunoprecipitated CSB from
CS1AN-sv cells reconstituted with CSB. Nothing was immuno-
precipitated from CS1AN-sv cells. The band marked with an
asterisk, present in lanes 1, 2 and 3, is of unknown identity;
however, this protein could not be immunoprecipitated with 1B1
and is, therefore, likely cross-reacting with the polyclonal antibody. (TIFF) Gene expression analysis Total RNA was prepared using TRIzol (Invitrogen). AMV
reverse transcriptase and random primers were used for first
strand cDNA synthesis (Roche). cDNA was analyzed by real-time
PCR using a MyiQ thermal cycler and SYBR green (BioRad). Expression was first normalized against b-actin and fold over
vector-only control was then calculated using DDCt method [57]. Primers used for RT-qPCR are listed in Table S6. ChIP-qPCR
assays were performed as previously described [12]. Primers used
for ChIP-qPCR are listed in Table S5. The Genomic Regions Enrichment of Annotations Tool
(GREAT, version 2.0.2) was used for pathway analysis of CSB
occupancy sites, using the ‘‘MSigDB pathways’ category’’ [59]. The assignment of peaks to genes was made using the following
parameters: proximal 5 kb upstream, 1 kb downstream, plus distal
up to 1000 kb. April 2014 | Volume 10 | Issue 4 | e1004284 PLOS Genetics | www.plosgenetics.org 14 C-Jun Targets CSB to Regulate Transcription seq peaks). Hypergeometric p-values were calculated against the
total number of CSB peaks for the c-Jun/Ap-1 motifs. (TIFF) Figure S5
Genomic distribution of CSB peaks containing TPA-
response element. Distributions were determined using the CEAS
package and are presented as a pie chart. The table includes p-
values for these distributions. (TIFF) Micrococcal nuclease digestion and mononucleosomal
DNA purification Figure S3
CSB and CSBDN1 occupancy positions relative to
transcription start sites. The number of CSB and CSBDN1
binding sites were plotted as a function of distance from
transcription start sites (TSS) using a bin size of 500 bp. (TIFF) Formaldehyde-fixed nuclei were isolated from each cell line as
described previously [58,60]. Nuclei (A260 = 500) were incubated
with MNase (final concentration 25 U/ml) at 37uC for 10 min and
reverse cross-linked at 65uC for 16 hours. After phenol-chloroform
extraction, digested DNA was resolved on a 1.2% agarose gel in
16TAE
at
100 V
for
3.5 hr. Mononucleosome-size
DNA
fragments were purified from gel slices and subjected to qPCR
analysis (Figure S6). Primers used in MNase-qPCR assays are
listed in Table S7. CSB peak classification according to chromatin features CSB peak classification according to chromatin features
CSB, top CSB (rpm greater than 1) and CSBDN1 peaks were
classified using the epigenomic information derived from NHLF
(normal human lung fibroblast) cells, via different chromatin
features using chromHMM [34,61]. The region annotation used
here is the result of unsupervised learning, finding an HMM with
15 states that minimizes the entropy of observed histone
modifications, and afterwards interpreted using prior biological
knowledge. Peaks were assigned to regions according to the
location of the peak centers. p-values were calculated using a one-
sided binomial test. (TIFF) Figure S4
Genomic distributions of peaks common or unique to
CSB and CSBDN1. Distributions were determined using the
CEAS package and are presented as pie charts. The table includes
p-values for these distributions. (TIFF) References 1. Nance MA, Berry SA (1992) Cockayne syndrome: review of 140 cases. Am J Med
Genet 42: 68–84. 23. Aamann MD, Sorensen MM, Hvitby C, Berquist BR, Muftuoglu M, et al. (2010) Cockayne syndrome group B protein promotes mitochondrial DNA
stability by supporting the DNA repair association with the mitochondrial
membrane. Faseb J 24: 2334–2346. 2. Lake RJ, Fan HY (2013) Structure, function and regulation of CSB: a multi-
talented gymnast. Mech Ageing Dev 134: 202–211. 24. Scheibye-Knudsen M, Croteau DL, Bohr VA (2013) Mitochondrial deficiency in
Cockayne syndrome. Mech Ageing Dev 134: 275–283. 3. Clapier CR, Cairns BR (2009) The biology of chromatin remodeling complexes. Annu Rev Biochem 78: 273–304. 25. Balajee AS, May A, Dianov GL, Friedberg EC, Bohr VA (1997) Reduced RNA
polymerase II transcription in intact and permeabilized Cockayne syndrome
group B cells. Proc Natl Acad Sci U S A 94: 4306–4311. 4. Narlikar GJ, Fan HY, Kingston RE (2002) Cooperation between complexes that
regulate chromatin structure and transcription. Cell 108: 475–487. 5. Torigoe SE, Patel A, Khuong MT, Bowman GD, Kadonaga JT (2013) ATP-
dependent chromatin assembly is functionally distinct from chromatin
remodeling. eLife 2: e00863. g
p
26. Bradsher J, Auriol J, Proietti de Santis L, Iben S, Vonesch JL, et al. (2002) CSB is
a component of RNA pol I transcription. Mol Cell 10: 819–829. a component of RNA pol I transcription. Mol Cell 10: 819–82 27. Ve´lez-Cruz R, Egly J-M (2013) Cockayne syndrome group B (CSB) protein: At
the crossroads of transcriptional networks. Mech Ageing Dev 134: 234–242. 6. Phelan ML, Sif S, Narlikar GJ, Kingston RE (1999) Reconstitution of a core
chromatin remodeling complex from SWI/SNF subunits. Mol Cell 3: 247–253. 28. Newman JC, Bailey AD, Weiner AM (2006) Cockayne syndrome group B
protein (CSB) plays a general role in chromatin maintenance and remodeling. Proc Natl Acad Sci USA 103: 9613–9618. 7. Wu JI, Lessard J, Olave IA, Qiu Z, Ghosh A, et al. (2007) Regulation of
dendritic development by neuron-specific chromatin remodeling complexes. Neuron 56: 94–108. 8. Cho I, Tsai PF, Lake RJ, Basheer A, Fan HY (2013) ATP-Dependent
Chromatin Remodeling by Cockayne Syndrome Protein B and NAP1-Like
Histone Chaperones Is Required for Efficient Transcription-Coupled DNA
Repair. PLoS Genet 9: e1003407. 29. Bailey AD, Gray LT, Pavelitz T, Newman JC, Horibata K, et al. References (2012) The
conserved Cockayne syndrome B-piggyBac fusion protein (CSB-PGBD3) affects
DNA repair and induces both interferon-like and innate antiviral responses in
CSB-null cells. DNA Repair (Amst) 11: 488–501. 9. Auble DT, Hansen KE, Mueller CG, Lane WS, Thorner J, et al. (1994) Mot1, a
global repressor of RNA polymerase II transcription, inhibits TBP binding to
DNA by an ATP-dependent mechanism. Genes Dev 8: 1920–1934. 30. Newman JC, Bailey AD, Fan HY, Pavelitz T, Weiner AM (2008) An abundant
evolutionarily conserved CSB-PiggyBac fusion protein expressed in Cockayne
syndrome. PLoS Genet 4: e1000031. y
31. Langmead B, Trapnell C, Pop M, Salzberg SL (2009) Ultrafast and memory-
efficient alignment of short DNA sequences to the human genome. Genome Biol
10: R25. 10. Selby CP, Sancar A (1997) Human transcription-repair coupling factor CSB/
ERCC6 is a DNA-stimulated ATPase but is not a helicase and does not disrupt
the ternary transcription complex of stalled RNA polymerase II. J Biol Chem
272: 1885–1890. 32. Shin H, Liu T, Manrai AK, Liu XS (2009) CEAS: cis-regulatory element
annotation system. Bioinformatics 25: 2605–2606. 11. Citterio E, Rademakers S, van der Horst GT, van Gool AJ, Hoeijmakers JH, et
al. (1998) Biochemical and biological characterization of wild-type and ATPase-
deficient Cockayne syndrome B repair protein. J Biol Chem 273: 11844–11851. 33. Consortium EP (2011) A user’s guide to the encyclopedia of DNA elements
(ENCODE). PLoS Biol 9: e1001046. 12. Lake RJ, Basheer A, Fan HY (2011) Reciprocally regulated chromatin
association of Cockayne syndrome protein B and p53 protein. J Biol Chem
286: 34951–34958. 34. Ernst J, Kheradpour P, Mikkelsen TS, Shoresh N, Ward LD, et al. (2011)
Mapping and analysis of chromatin state dynamics in nine human cell types. Nature 473: 43–49. 35. Lake RJ, Geyko A, Hemashettar G, Zhao Y, Fan HY (2010) UV-induced
association of the CSB remodeling protein with chromatin requires ATP-
dependent relief of N-terminal autorepression. Mol Cell 37: 235–246. 13. Citterio E, Van Den Boom V, Schnitzler G, Kanaar R, Bonte E, et al. (2000)
ATP-dependent chromatin remodeling by the Cockayne syndrome B DNA
repair-transcription-coupling factor. Mol Cell Biol 20: 7643–7653. 14. Beerens N, Hoeijmakers JH, Kanaar R, Vermeulen W, Wyman C (2005) The
CSB protein actively wraps DNA. J Biol Chem 280: 4722–4729. 36. Mallery DL, Tanganelli B, Colella S, Steingrimsdottir H, van Gool AJ, et al. (1998) Molecular analysis of mutations in the CSB (ERCC6) gene in patients
with Cockayne syndrome. References Am J Hum Genet 62: 77–85. 15. Yang JG, Madrid TS, Sevastopoulos E, Narlikar GJ (2006) The chromatin-
remodeling enzyme ACF is an ATP-dependent DNA length sensor that
regulates nucleosome spacing. Nat Struct Mol Biol 13: 1078–1083. 37. Eferl R, Wagner EF (2003) AP-1: a double-edged sword in tumorigenesis. Nat
Rev Cancer 3: 859–868. 16. He X, Fan HY, Garlick JD, Kingston RE (2008) Diverse Regulation of SNF2h
Chromatin Remodeling by Noncatalytic Subunits. Biochemistry 47: 7025–7033. 38. Shaulian E, Karin M (2002) AP-1 as a regulator of cell life and death. Nat Cell
Biol 4: E131–136. 39. Gray LT, Fong KK, Pavelitz T, Weiner AM (2012) Tethering of the Conserved
piggyBac Transposase Fusion Protein CSB-PGBD3 to Chromosomal AP-1
Proteins Regulates Expression of Nearby Genes in Humans. PLoS Genet 8:
e1002972. 17. Kristensen U, Epanchintsev A, Rauschendorf MA, Laugel V, Stevnsner T, et al. (2013) Regulatory interplay of Cockayne syndrome B ATPase and stress-
response gene ATF3 following genotoxic stress. Proc Natl Acad Sci U S A 110:
E2261–2270. 18. Hanawalt PC, Spivak G (2008) Transcription-coupled DNA repair: two decades
of progress and surprises. Nat Rev Mol Cell Biol 9: 958–970. 40. Askarian-Amiri ME, Crawford J, French JD, Smart CE, Smith MA, et al. (2011)
SNORD-host RNA Zfas1 is a regulator of mammary development and a
potential marker for breast cancer. RNA 17: 878–891. 19. Troelstra C, van Gool A, de Wit J, Vermeulen W, Bootsma D, et al. (1992)
ERCC6, a member of a subfamily of putative helicases, is involved in
Cockayne’s syndrome and preferential repair of active genes. Cell 71: 939–953. 41. Jackson AP, Eastwood H, Bell SM, Adu J, Toomes C, et al. (2002) Identification
of microcephalin, a protein implicated in determining the size of the human
brain. Am J Hum Genet 71: 136–142. 20. Fousteri M, Vermeulen W, van Zeeland AA, Mullenders LH (2006) Cockayne
syndrome A and B proteins differentially regulate recruitment of chromatin
remodeling and repair factors to stalled RNA polymerase II in vivo. Mol Cell 23:
471–482. 42. Talkowski ME, Rosenfeld JA, Blumenthal I, Pillalamarri V, Chiang C, et al. (2012) Sequencing chromosomal abnormalities reveals neurodevelopmental loci
that confer risk across diagnostic boundaries. Cell 149: 525–537. 21. Kyng KJ, May A, Brosh RM, Jr., Cheng WH, Chen C, et al. (2003) The
transcriptional response after oxidative stress is defective in Cockayne syndrome
group B cells. Oncogene 22: 1135–1149. 43. Table S6
Primers used in RT-qPCR assays.
(DOCX) Table S7
Primers used in MNase-qPCR assays. (DOCX) Table S7
Primers used in MNase-qPCR assays. (DOCX) Table S2
Genomic annotation of peaks unique to CSB. (XLSX) Author Contributions Table S3
Common sites of CSB and CSB-PGBD3 occupancy. (XLSX) Conceived and designed the experiments: RJL ELB PFT HYF. Performed
the experiments: RJL ELB PFT HYF. Analyzed the data: RJL ELB PFT
HYF JJ IC KJW. Contributed reagents/materials/analysis tools: JJ IC
KJW. Wrote the paper: RJL ELB HYF. Drafted parts of the paper: PFT JJ
KJW. Conceived and designed the experiments: RJL ELB PFT HYF. Performed
the experiments: RJL ELB PFT HYF. Analyzed the data: RJL ELB PFT
HYF JJ IC KJW. Contributed reagents/materials/analysis tools: JJ IC
KJW. Wrote the paper: RJL ELB HYF. Drafted parts of the paper: PFT JJ
KJW. Conceived and designed the experiments: RJL ELB PFT HYF. Performed
the experiments: RJL ELB PFT HYF. Analyzed the data: RJL ELB PFT Table S4
Lists of genes correlated in Tables 5 and 6. (XLSX) Table S4
Lists of genes correlated in Tables 5 and 6. (XLSX) Table S4
Lists of genes correlated in Tables 5 and 6. (XLSX) KJW. Wrote the paper: RJL ELB HYF. Drafted parts of the paper: PFT JJ
KJW. Table S5
Primers used in ChIP-qPCR assays. (DOCX) Table S5
Primers used in ChIP-qPCR assays. (DOCX) Comparisons of CSB ChIP-seq data with transcription
profiling data To compare the CSB ChIP-seq data with the microarray data,
we used the lists of up- and down-regulated genes generated by
Newman et al. (2006) and Bailey et al. (2012) [28,29]. Newman et al. identified CSB-responsive genes in hTERT immortalized CS1AN
cells. Bailey et al. (2012) identified CSB-responsive, PGBD3-
responsive, and CSB+PGBD3-responsive genes in USS1KO cells. We determined the number of genes whose body or promoter
regions overlapped with CSB peaks. p-values were calculated after
randomly generating 17,779 peaks (the same number of CSB ChIP- April 2014 | Volume 10 | Issue 4 | e1004284 PLOS Genetics | www.plosgenetics.org 15 C-Jun Targets CSB to Regulate Transcription Table S1
Genomic annotation of peaks common to CSB and
CSBDN1. (XLSX) Table S1
Genomic annotation of peaks common to CSB and
CSBDN1. (XLSX)
Table S2
Genomic annotation of peaks unique to CSB. (XLSX)
Table S3
Common sites of CSB and CSB-PGBD3 occupancy. (XLSX)
Table S4
Lists of genes correlated in Tables 5 and 6. (XLSX)
Table S5
Primers used in ChIP-qPCR assays. (DOCX) Table S6
Primers used in RT-qPCR assays. (DOCX) C-Jun Targets CSB to Regulate Transcription 45. Osaki M, Inoue T, Yamaguchi S, Inaba A, Tokuyasu N, et al. (2007) MAD1
(mitotic arrest deficiency 1) is a candidate for a tumor suppressor gene in human
stomach. Virchows Arch 451: 771–779. 54. Yuan X, Feng W, Imhof A, Grummt I, Zhou Y (2007) Activation of RNA
polymerase I transcription by cockayne syndrome group B protein and histone
methyltransferase G9a. Mol Cell 27: 585–595. 46. Liu Y, Boukhelifa M, Tribble E, Morin-Kensicki E, Uetrecht A, et al. (2008) The
Sac1 phosphoinositide phosphatase regulates Golgi membrane morphology and
mitotic spindle organization in mammals. Mol Biol Cell 19: 3080–3096. 55. Berquist BR, Canugovi C, Sykora P, Wilson DM, 3rd, Bohr VA (2012) Human
Cockayne syndrome B protein reciprocally communicates with mitochondrial
proteins and promotes transcriptional elongation. Nucleic Acids Res 40: 8392–
8405. 47. Tee WW, Pardo M, Theunissen TW, Yu L, Choudhary JS, et al. (2010) Prmt5 is
essential for early mouse development and acts in the cytoplasm to maintain ES
cell pluripotency. Genes Dev 24: 2772–2777. 56. Brooks PJ, Cheng TF, Cooper L (2008) Do all of the neurologic diseases in
patients with DNA repair gene mutations result from the accumulation of DNA
damage? DNA Repair (Amst) 7: 834–848. p
p
y
48. Maserati M, Walentuk M, Dai X, Holston O, Adams D, et al. (2011) Wdr74 is
required for blastocyst formation in the mouse. PLoS One 6: e22516. 57. Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the
comparative CT method. Nat Protoc 3: 1101–1108. 49. Yao TW, Kim WS, Yu DM, Sharbeen G, McCaughan GW, et al. (2011) A
novel role of dipeptidyl peptidase 9 in epidermal growth factor signaling. Mol
Cancer Res 9: 948–959. 58. Dennis JH, Fan HY, Reynolds SM, Yuan G, Meldrim JC, et al. (2007)
Independent and complementary methods for large-scale structural analysis of
mammalian chromatin. Genome Res 17: 928–939. 50. Yadon AN, Singh BN, Hampsey M, Tsukiyama T (2013) DNA looping
facilitates targeting of a chromatin remodeling enzyme. Mol Cell 50: 93–103. 59. McLean CY, Bristor D, Hiller M, Clarke SL, Schaar BT, et al. (2010) GREAT
improves functional interpretation of cis-regulatory regions. Nat Biotechnol 28:
495–501. 51. Zhao W, Wang L, Zhang M, Wang P, Zhang L, et al. (2011) NF-kappaB- and
AP-1-mediated DNA looping regulates osteopontin transcription in endotoxin-
stimulated murine macrophages. J Immunol 186: 3173–3179. 60. References Iijima K, Yamada H, Miharu M, Imadome K, Miyagawa Y, et al. (2012)
ZNF385B is characteristically expressed in germinal center B cells and involved
in B-cell apoptosis. Eur J Immunol 42: 3405–3415. 44. Petroziello J, Yamane A, Westendorf L, Thompson M, McDonagh C, et al. (2004) Suppression subtractive hybridization and expression profiling identifies a
unique set of genes overexpressed in non-small-cell lung cancer. Oncogene 23:
7734–7745. 22. Tuo J, Jaruga P, Rodriguez H, Bohr VA, Dizdaroglu M (2003) Primary
fibroblasts of Cockayne syndrome patients are defective in cellular repair of 8-
hydroxyguanine and 8-hydroxyadenine resulting from oxidative stress. Faseb J
17: 668–674. April 2014 | Volume 10 | Issue 4 | e1004284 April 2014 | Volume 10 | Issue 4 | e1004284 PLOS Genetics | www.plosgenetics.org 16 C-Jun Targets CSB to Regulate Transcription Fragoso G, Hager GL (1997) Analysis of in vivo nucleosome positions by
determination of nucleosome-linker boundaries in crosslinked chromatin. Methods 11: 246–252. 52. Yadon AN, Van de Mark D, Basom R, Delrow J, Whitehouse I, et al. (2010)
Chromatin remodeling around nucleosome-free regions leads to repression of
noncoding RNA transcription. Mol Cell Biol 30: 5110–5122. 61. Ernst J, Kellis M (2012) ChromHMM: automating chromatin-state discovery
and characterization. Nat Meth 9: 215–216. g
p
53. Mueller-Planitz F, Klinker H, Becker PB (2013) Nucleosome sliding mecha-
nisms: new twists in a looped history. Nat Struct Mol Biol 20: 1026–1032. April 2014 | Volume 10 | Issue 4 | e1004284 PLOS Genetics | www.plosgenetics.org 17
|
https://openalex.org/W3102561211
|
https://www.aanda.org/articles/aa/pdf/2020/06/aa38054-20.pdf
|
English
| null |
Connecting measurements of solar and stellar brightness variations
|
Astronomy & astrophysics
| 2,020
|
cc-by
| 9,283
|
1. Introduction reviews), a comparison between solar and stellar brightness mea-
surements is far from straightforward (see e.g. Basri et al. 2010;
Reinhold et al. 2020; Witzke et al. 2020). Firstly, solar and stel-
lar brightness variations have been measured in different spectral
passbands. Because the amplitude of the solar rotational vari-
ability strongly depends on the wavelength (Solanki et al. 2013;
Ermolli et al. 2013), the solar and stellar brightness records can
be reliably compared only after conversion from one passband
to another. Secondly, the solar brightness variations have (so far)
only been measured from the ecliptic plane, which is very close
to the solar equatorial plane (the angle between the solar equa-
tor and ecliptic plane is about 7.25◦). The values of the angle
between the line of sight of the observer and the rotation axes of
the observed stars (hereinafter referred to as the inclination) are
mostly unknown. Dedicated planet-hunting photometric missions such as CoRoT
(Convection, Rotation and planetary Transit, see Baglin et al. 2006; Bordé et al. 2003), Kepler (Borucki et al. 2010), and
TESS (Transiting Exoplanet Survey Satellite, see Ricker et al. 2014), and also the Gaia space observatory (Gaia Collaboration
2016) have made it possible to measure stellar brightness vari-
ability with unprecedented precision. In particular, they allow
studying stellar brightness variations caused by transits (as the
star rotates) and the evolution of magnetic features, that is, bright
faculae and dark spots. Such variations are often referred to as
rotational stellar variability. The plethora of stellar observational
data rekindled an interest in the questions of how typical our Sun
is as an active star, and more specifically, how the solar rotational
variability compares to that of solar-like stars. Furthermore,
these data allow probing whether the solar activity paradigm
is also valid for other stars. This requires comparing the stel-
lar properties and behaviour with those of the Sun. While the
solar variability has been measured for more than four decades
now by various dedicated space missions (see e.g. Fröhlich
2012; Ermolli et al. 2013; Solanki et al. 2013; Kopp 2016, for Studies comparing solar and stellar rotational brightness
variations have used different types of solar brightness mea-
surements. Reinhold et al. (2020), for instance, used the total
solar irradiance (TSI), that is, the solar radiative flux at 1 AU
integrated over all wavelengths. More commonly, however, the
solar variability was characterised (see e.g. Basri et al. ABSTRACT Context. A comparison of solar and stellar brightness variations is hampered by the difference in spectral passbands that are used in
observations, and also by the possible difference in the inclination of the solar and stellar rotation axes from the line of sight. Context. A comparison of solar and stellar brightness variations is hampered by the difference in spectral passbands that are used in
observations, and also by the possible difference in the inclination of the solar and stellar rotation axes from the line of sight. Aims. We calculate the rotational variability of the Sun as it would be measured in passbands used for stellar observations. In partic-
ular, we consider the filter systems used by the CoRoT, Kepler, TESS, and Gaia space missions. We also quantify the effect of the
inclination of the rotation axis on the solar rotational variability. Aims. We calculate the rotational variability of the Sun as it would be measured in passbands used for stellar observations. In partic-
ular, we consider the filter systems used by the CoRoT, Kepler, TESS, and Gaia space missions. We also quantify the effect of the
inclination of the rotation axis on the solar rotational variability. y
Methods. We employed the spectral and total irradiance reconstruction (SATIRE) model to calculate solar brightness variations in
different filter systems as observed from the ecliptic plane. We then combined the simulations of the surface distribution of the mag-
netic features at different inclinations using a surface flux transport model with the SATIRE calculations to compute the dependence
of the variability on the inclination. Methods. We employed the spectral and total irradiance reconstruction (SATIRE) model to calculate solar brightness variations in
different filter systems as observed from the ecliptic plane. We then combined the simulations of the surface distribution of the mag-
netic features at different inclinations using a surface flux transport model with the SATIRE calculations to compute the dependence
of the variability on the inclination. Results. For an ecliptic-bound observer, the amplitude of the solar rotational variability, as observed in the total solar irradiance
(TSI), is 0.68 mmag (averaged over solar cycles 21–24). We obtained corresponding amplitudes in the Kepler (0.74 mmag), CoRoT
(0.73 mmag), TESS (0.62 mmag), Gaia G (0.74 mmag), Gaia GRP (0.62 mmag), and Gaia GBP (0.86 mmag) passbands. Decreasing
the inclination of the rotation axis decreases the rotational variability. ABSTRACT For a sample of randomly inclined stars, the variability is on
average 15% lower in all filter systems we considered. This almost compensates for the difference in amplitudes of the variability in
TSI and Kepler passbands, making the amplitudes derived from the TSI records an ideal representation of the solar rotational vari-
ability for comparison to Kepler stars with unknown inclinations. Conclusions. The TSI appears to be a relatively good measure of solar variability for comparisons with stellar measurements in the
CoRoT, Kepler, TESS Gaia G, and Gaia GRP filters. Whereas the correction factors can be used to convert the variability amplitude
from solar measurements into the values expected for stellar missions, the inclination affects the shapes of the light curves so that a
much more sophisticated correction than simple scaling is needed to obtain light curves out of the ecliptic for the Sun. Key words. Sun: activity – stars: solar-type – stars: rotation – Sun: rotation Astronomy
&
Astrophysics Astronomy
&
Astrophysics Astronomy
&
Astrophysics A&A 638, A56 (2020)
https://doi.org/10.1051/0004-6361/202038054
c⃝N.-E. Nèmec et al. 2020 A&A 638, A56 (2020)
https://doi.org/10.1051/0004-6361/202038054
c⃝N.-E. Nèmec et al. 2020 Connecting measurements of solar and stellar brightness
variations N.-E. Nèmec1, E. I¸sık2,1, A. I. Shapiro1, S. K. Solanki1,3, N. A. Krivova1, and Y. Unruh 1 Max-Planck-Institut für Sonnensystemforschung, Justus-von-Liebig-Weg 3, 37077 Göttingen, Germany
e-mail: nemec@mps.mpg.de p
pg
2 Dept. of Computer Science, Turkish-German University, ¸Sahinkaya Cd. 108, 34820 Beykoz, Istanbul, Turkey
3 School of Space Research, Kyung Hee University, Yongin, Gyeonggi 446-701, Korea
4 Imperial College, Astrophysics Group, Blackett Laboratory, London SW7 2BZ, UK Received 30 March 2020 / Accepted 14 April 2020 Received 30 March 2020 / Accepted 14 April 2020 Open Access article, published by EDP Sciences, under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0),
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Open Access funding provided by Max Planck Society. 2.2. Filter systems In this section we multiply the SATIRE-S SSI output with the
response function of a given filter and integrate it over the entire
filter passband to obtain the solar light curve in the correspond-
ing filter. It is important to take the nature of the detectors used
in different instruments into account (see e.g. Maxted 2018). In particular, while solar instruments (e.g. VIRGO/SPM and all
TSI instruments) measure the energy of the incoming radiation,
charge-coupled devices (CCDs) used in Kepler, Gaia, and TESS
count the number of photons and not their energy. In order to
obtain the solar light curve, LC, as it would be measured by the
instrument counting photons, we therefore follow In this paper we seek to overcome these two hurdles and
quantify solar variability in passbands that are used by different
stellar space missions and at different inclinations. In Sect. 2 we
employ the spectral and total irradiance reconstruction (SATIRE;
Fligge et al. 2000; Krivova et al. 2003) model of solar brightness
variations to show how the actual solar brightness variations are
related to solar brightness variations as they would be observed
in spectral passbands used by stellar missions. We also estab-
lish the connection between the TSI and VIRGO/SPM measure-
ments. In Sect. 3 we follow the approach developed by N20 to
quantify the effect of the inclination on the brightness variations. We discuss how the Sun as observed by Kepler can be modelled
using light curves obtained by VIRGO/SPM in Sect. 4 before we
summarise our results and draw conclusions in Sect. 5. LC =
λ2
Z
λ1
R(λ) · I(λ) λ
h · c dλ,
(1) =
λ2
Z
R(λ) · I(λ) λ
h · c dλ,
(1) (1) where λ1 and λ2 are the blue and red threshold wavelengths of
the filter passband, R(λ) is the response function of the filter,
and I(λ) is the spectral irradiance at a given wavelength, h is the
Planck constant, and c the speed of light. First we consider several broad-band filters used by the
planet-hunting missions: CoRoT, Kepler, and TESS. The spec-
tral passbands employed in these missions are shown in the top
panel of Fig. 1, along with the quiet-Sun spectrum calculated by
Unruh et al. (1999) and used in SATIRE-S. Clearly, the CoRoT
and Kepler response functions are very similar to each other
because both missions focused on G stars. 1. Introduction Because of its almost equator-on view,
the Sun would therefore appear on average more variable than
stars with the same activity level that are observed at random
inclinations. At the same time, an easy-to-use receipt for cor-
recting the variability for the inclination effect is lacking so far,
and consequently, the inclination has not yet been quantitatively
accounted for in solar-stellar comparison studies. ing four solar cycles. As especially the early ground-based obser-
vations contain gaps in the data, we used the SATIRE-S model
as presented by Yeo et al. (2014) (version 20190621), where
the gaps in spectral solar irradiance (SSI) and TSI have been
filled using the information provided by solar activity indices. SATIRE-S was shown to reproduce the apparent variability of
the Sun as observed, in both the SSI and in the TSI (see Ball et al. 2012, 2014; Yeo et al. 2014; Danilovic et al. 2016, and refer-
ences therein). The spectral resolution of the SATIRE output is
1 nm below 290 nm, 2 nm between 290 nm and 999 nm, and 5 nm
above 1000 nm. This is fully sufficient for the calculations pre-
sented in this study. 2.1. SATIRE-S The SATIRE model Fligge et al. (2000), Krivova et al. (2003)
attributes the brightness variations of the Sun on timescales
longer than a day to the presence of magnetic features on its
surface, such as bright faculae and dark spots. The two main
building blocks of SATIRE are the areas and the positions of
the magnetic features on the solar disc as well as contrasts of
these features relative to the quiet Sun (i.e. regions on the solar
surface free from any apparent manifestations of magnetic activ-
ity). The contrasts of the magnetic features as a function of disc
position and wavelength were computed by Unruh et al. (1999)
with the spectral synthesis block of the ATLAS9 code (Kurucz
1992; Castelli & Kurucz 1994). The 1D atmospheric structure
of the two spot components (umbra and penumbra) and of the
quiet Sun were calculated using radiative equilibrium models
produced with the ATLAS9 code, while the facular model is a
modified version of FAL-P by Fontenla et al. (1993). Gaia measures stellar brightness in three different chan-
nels (Gaia Collaboration 2016). Gaia G is sensitive to photons
between 350 and 1000 nm. Additionally, two prisms disperse the
incoming light between 330 and 680 nm for the Blue Photometer
(hereafter referred to as Gaia GBP) and between 640–1050 nm
for the Red Photometer (hereafter, referred to as Gaia GRP). The
response functions are shown in the middle panel in Fig. 1. We
employ the revised passbands used for the second data release of
Gaia (Gaia DR2, Evans et al. 2018) for the calculations. Solar-stellar comparison studies have often used the solar
variability as measured by the VIRGO/SPM instrument. SPM
comprises three photometers, with a bandwidth of 5 nm oper-
ating at 402 nm (blue), 500 nm (green), and 862 nm (red). The
response functions are shown in Fig. 1 in the bottom panel. We
refer to these filters from now on as VIRGO-blue, -green, and
-red. Various versions of the SATIRE model exist. In this section
we employ the most precise version, which is SATIRE-S, where
the suffix “S” stands for the satellite era (Ball et al. 2014;
Yeo et al. 2014). SATIRE-S uses the distribution of magnetic
features on the solar disc obtained from observed magnetograms
and continuum disc images and spans from 1974 to today, cover- 1. Introduction 2010;
Gilliland et al. 2011; Harrison et al. 2012) using measurements A56, page 1 of 8 A&A 638, A56 (2020) by the Variability of solar IRradiance and Gravity Oscilla-
tions/Sun PhotoMeters (VIRGO/SPM) (Fröhlich et al. 1995,
1997) instrument on board the Solar and Heliospheric Observa-
tory (SoHO). VIRGO/SPM measures solar brightness in three
filters with a bandwidth of 5 nm each. Neither VIRGO/SPM
nor TSI measurements can be directly compared to records of
stellar brightness variability, which typically cover wavelength
ranges broader than the VIRGO/SPM filters, but much narrower
than the TSI. Accurate estimations of solar variability in pass-
bands used for stellar measurements have therefore so far been
missing. Some effort has previously been made to model the
solar rotational variability as it would be observed out of ecliptic
(e.g. Vieira et al. 2012; Shapiro et al. 2016; Nèmec et al. 2020). In particular, Shapiro et al. (2016) and Nèmec et al. (2020)
(hereinafter N20) have shown that the amplitude of the solar
brightness variations on the rotational timescale decreases with
decreasing inclination. Because of its almost equator-on view,
the Sun would therefore appear on average more variable than
stars with the same activity level that are observed at random
inclinations. At the same time, an easy-to-use receipt for cor-
recting the variability for the inclination effect is lacking so far,
and consequently, the inclination has not yet been quantitatively
accounted for in solar-stellar comparison studies. by the Variability of solar IRradiance and Gravity Oscilla-
tions/Sun PhotoMeters (VIRGO/SPM) (Fröhlich et al. 1995,
1997) instrument on board the Solar and Heliospheric Observa-
tory (SoHO). VIRGO/SPM measures solar brightness in three
filters with a bandwidth of 5 nm each. Neither VIRGO/SPM
nor TSI measurements can be directly compared to records of
stellar brightness variability, which typically cover wavelength
ranges broader than the VIRGO/SPM filters, but much narrower
than the TSI. Accurate estimations of solar variability in pass-
bands used for stellar measurements have therefore so far been
missing. Some effort has previously been made to model the
solar rotational variability as it would be observed out of ecliptic
(e.g. Vieira et al. 2012; Shapiro et al. 2016; Nèmec et al. 2020). In particular, Shapiro et al. (2016) and Nèmec et al. (2020)
(hereinafter N20) have shown that the amplitude of the solar
brightness variations on the rotational timescale decreases with
decreasing inclination. A56, page 2 of 8 2.2. Filter systems TESS is designed to
observe cooler stars than Kepler, hence the response function is
shifted towards the red part of the spectrum. 2.3. Results 3 we compare the R30 values for all the filter systems
introduced in Sect. 2.2 to R30 of the TSI for a 1000-day inter-
val starting 26 July 2013. This interval therefore includes the
maximum of solar 24 as well. To better quantify the dependence
of R30 on the passband, we show linear regressions between
the variability R30 in each filter system and the TSI in Fig. 4. The slopes of the linear regressions are listed in Table 1. The
Pearson correlation coefficient is above 0.98 for all of the fil-
ter systems. The slope of the linear regressions depends on the
filter system that is considered. For example, TESS and Gaia
G regressions have a slope close to 1, but GBP displays a slope
>1, whereas VIRGO-red exhibits a slope <1. As expected, the
slope is highest for the blue VIRGO filter, where the amplitude
of the variability is highest. We note that the good agreement of
the TSI with the red filters is expected to be valid only for the
rotational variability, which is dominated by spots. In contrast,
the solar irradiance variability on the activity cycle timescale is
given by the delicate balance between facular and spot compo-
nents, and consequently has a very sophisticated spectral profile
(Shapiro et al. 2016; Witzke et al. 2018). Thus, values of slopes
from Table 1 cannot be extrapolated from rotational to activity
cycle timescales (see Shapiro et al. 2016, for the detailed discus-
sion). 200
300
400
500
600
700
800
900
1000 1100 1200
Wavelenght [ m]
0.0
0.2
0.4
0.6
0.8
1.0
1.2
Tra smissivity
VIRGO/blue
VIRGO/gree
VIRGO/red
0.0
0.5
1.0
1.5
2.0
Irradia ce [Wm−2 m−1] 5
0
5
0
Irradia ce [Wm−2 m−1] Table 2 lists the cycle-averaged values of R30 for all pass-
bands in mmag. Together, Fig. 4, and Tables 1 and 2 show that
the TSI is a passable representative for the variability on the solar
rotation timescale as it would be observed in the TESS, Kepler,
CoRoT, Gaia G, Gaia GRP, and VIRGO-red filters, but it notice-
ably underestimates the variability in GBP, VIRGO-green, and
VIRGO-blue. Fig. 1. Response functions for the various filter systems used in this
study. For comparison, the quiet-Sun spectrum used by SATIRE-S is
plotted in grey in each panel. Top panel: Kepler, TESS, and CoRoT. Middle panel: three Gaia passbands. Bottom panel: three VIRGO/SPM
channels. 2.3. Results Figure 2 shows the solar light curve for the period of 2456700–
2456850 JD (24 February 2014 – 11 July 2014) as it would be A56, page 2 of 8 N.-E. Nèmec et al.: Connecting solar and stellar variabilities the narrow VIRGO-blue filter show far stronger variability than
the TSI. 200
300
400
500
600
700
800
900
1000 1100 1200
Wavelenght [nm]
0.0
0.2
0.4
0.6
0.8
1.0
1.2
Transmissivity
Kepler
C R T
TESS
0.0
0.5
1.0
1.5
2.0
Irradiance [Wm−2nm−1]
200
300
400
500
600
700
800
900
1000 1100 1200
Wavelenght [nm]
0.0
0.2
0.4
0.6
0.8
1.0
1.2
Transmissivit
GBP
G
GRP
0.0
0.5
1.0
1.5
2.0
Irradiance [Wm−2nm−1]
200
300
400
500
600
700
800
900
1000 1100 1200
Wavelenght [ m]
0.0
0.2
0.4
0.6
0.8
1.0
1.2
Tra smissivity
VIRGO/blue
VIRGO/gree
VIRGO/red
0.0
0.5
1.0
1.5
2.0
Irradia ce [Wm−2 m−1]
Fig. 1. Response functions for the various filter systems used in this
study. For comparison, the quiet-Sun spectrum used by SATIRE-S is
plotted in grey in each panel. Top panel: Kepler, TESS, and CoRoT. Middle panel: three Gaia passbands. Bottom panel: three VIRGO/SPM
channels. 200
300
400
500
600
700
800
900
1000 1100 1200
Wavelenght [nm]
0.0
0.2
0.4
0.6
0.8
1.0
1.2
Transmissivity
Kepler
C R T
TESS
0.0
0.5
1.0
1.5
2.0
Irradiance [Wm−2nm−1] To quantify the rotational variability, we computed the R30
values (see e.g. Basri et al. 2013). To do this, the obtained light
curves were split into 30-day segments, and within each seg-
ment, we calculated the difference between the extrema and
divided this value by the mean flux in the segment to derive the
relative variability. For the SATIRE-S time series, we directly
considered the difference between the extrema instead of the
differences between the 95th and 5th percentiles of sorted flux
values, as is usually done in the literature with the more noisy
Kepler measurements. We calculated R30 values for the period
1974–2019 (i.e. cycles 22–24). This allowed us to quantify the
mean level of solar variability in R30 that represents the full four
decades of TSI measurements. 0
600
700
800
9
Wavelenght [nm] 200
300
400
500
600
700
800
900
1000 1100 1200
Wavelenght [nm]
0.0
0.2
0.4
0.6
0.8
1.0
1.2
Transmissivit
GBP
G
GRP
0.0
0.5
1.0
1.5
2.0
Irradiance [Wm−2nm−1] In Fig. 2.3. Results Several studies (see e.g. Basri et al. 2010; Harrison et al. 2012) have assumed that the amplitude of the rotational solar
variability as it would be measured by Kepler is very close
to the amplitude calculated for the combined green and red
VIRGO/SPM light curves (in the following VIRGO/g+r). Here
we test this hypothesis. The variability R30 for Kepler compared
to VIRGO/g+r is shown in Fig. 5, which is limited to the same
time interval as Fig. 3. The two curves are remarkably similar to
one another. To test the similarity quantitatively, we show the lin-
ear regression of R30 between Kepler and VIRGO/g+r for four
solar cycles (21–24) in Fig. 6. The Pearson correlation coeffi-
cient is very high (0.999) and the slope deviates by only +0.8%
of unity averaged over four solar cycles. While these calcula-
tions are related to the amplitude of the rotational variability, R30,
we additionally calculate regressions between Kepler and Virgo
light curves in Sect. 4. We also directly connect the TSI and
VIRGO/g+r rotational variability. The linear regression between
R30 in TSI and VIRGO/g+r results in a slope of 0.88 (±0.002)
and a Pearson correlation coefficient of 0.995. observed in different passbands. This corresponds to a 150-day
interval during the maximum of cycle 24. This interval was cho-
sen arbitrarily to display the effect of the filter systems on the
solar variability. For this, we first divided each light curve in
90-day segments. This time span corresponds to Kepler quarters. This is motivated by the way Kepler observations are gathered
and reduced. We note that the detrending by the Kepler opera-
tional mode is applied here for purely illustrative purposes and
was not used for the calculations presented below. Within each
segment, we subtracted the mean value from the fluxes before
dividing the corresponding values by the mean flux in each seg-
ment. In all stellar broad-band filters, the light curve is remark-
ably similar in shape to the TSI (solid black curve), although the
amplitude can differ. As might be expected, the difference in the
amplitude of the variability is somewhat more conspicuous in
the blue filters. The Gaia GRP light curve is basically identical to
the TSI light curve, whereas the variability in Gaia GBP and in observed in different passbands. This corresponds to a 150-day
interval during the maximum of cycle 24. 3. Correction for the inclination
3.1. Approach then fed into the SATIRE model to calculate solar brightness
variations as they would be seen at different inclinations. A56, page 4 of 8 2.3. Results This interval was cho-
sen arbitrarily to display the effect of the filter systems on the
solar variability. For this, we first divided each light curve in
90-day segments. This time span corresponds to Kepler quarters. This is motivated by the way Kepler observations are gathered
and reduced. We note that the detrending by the Kepler opera-
tional mode is applied here for purely illustrative purposes and
was not used for the calculations presented below. Within each
segment, we subtracted the mean value from the fluxes before
dividing the corresponding values by the mean flux in each seg-
ment. In all stellar broad-band filters, the light curve is remark-
ably similar in shape to the TSI (solid black curve), although the
amplitude can differ. As might be expected, the difference in the
amplitude of the variability is somewhat more conspicuous in
the blue filters. The Gaia GRP light curve is basically identical to
the TSI light curve, whereas the variability in Gaia GBP and in A56, page 3 of 8 A&A 638, A56 (2020) A&A 638, A56 (2020)
-1000
-500
0
500
1000
Kepler
CoRoT
TESS
-1000
-500
0
500
1000
Normalised flux [ppm]
GAIA GBP
GAIA G
GAIA GRP
0
50
100
150
Time [days]
-1000
-500
0
500
1000
VIRGO/blue
0
50
100
150
Time [days]
VIRGO/green
0
50
100
150
Time [days]
VIRGO/red
Fig. 2. Normalised fluxes for the different filter systems compared to the TSI (black solid line). Top panels: Kepler, CoRoT, and TESS. Middle
panels: three Gaia passbands. Bottom panels: three VIRGO/SPM channels. Fig. 2. Normalised fluxes for the different filter systems compared to the TSI (black solid line). Top panels: Kepler, CoRoT, and TESS. Middle
panels: three Gaia passbands. Bottom panels: three VIRGO/SPM channels. 0
500
1000
1500
2000
2500
3000
Kepler
CoRoT
TESS
0
500
1000
1500
2000
2500
3000
R30 [ppm]
Gaia GBP
Gaia G
Gaia GRP
0
250
500
750
1000
Time [days]
0
500
1000
1500
2000
2500
3000
VIRGO/blue
0
250
500
750
1000
Time [days]
VIRGO/green
0
250
500
750
1000
Time [days]
VIRGO/red
Fig. 3. R30 in different filter systems compared to the TSI (black solid line) for a time span of 1000 days over solar cycle 22. Top panels: Kepler,
CoRoT, and TESS. Middle panels: three Gaia passbands. Bottom panels: three VIRGO/SPM channels. See the main text for the definition of R30. Fig. 3. 2.3. Results R30 in different filter systems compared to the TSI (black solid line) for a time span of 1000 days over solar cycle 22. Top panels: Kepler,
CoRoT, and TESS. Middle panels: three Gaia passbands. Bottom panels: three VIRGO/SPM channels. See the main text for the definition of R30. 3.1. Approach Such a randomisation is needed to ensure that the near
and far side of the Sun have on average equal activity, which is
a necessary condition for reliable calculations of the inclination
effect. As a result, our calculations reproduce the statistical prop-
erties of a given solar cycle, but they do not represent the actual
observed BMR emergences for that specific cycle. We stress that
in N20 we developed the model outlined above to study the effect
of the inclination on the power spectra of solar brightness varia-
tions. Here we use this model to explicitly study the dependence
of the variability amplitude on the rotational timescale and its
dependence on the inclination in different filters. we divided the time series for cycle 23 into 90-day segments,
which correspond to Kepler quarters. Within each quarter, we
de-trended the light curves. 90◦corresponds to an ecliptic-bound
observer, 57◦represents a weighted mean value of the inclina-
tion with weights equal to the probability of observing a given
inclination (sin(i)) for the inclination i), and 0◦corresponds to an
observer facing the north pole. We additionally show 30◦as an
intermediate point between 57◦and 0◦. For the inclination values
of 30◦, 57◦, and 90◦, the variability is brought about by the solar
rotation, as well as the emergence and evolution of magnetic fea-
tures. A polar-bound observer does not observe the rotational
modulation because there is no transit of magnetic features and
the variability is merely generated by their emergence and evo-
lution (see Nèmec et al. 2020, for further details). We empha-
sise just the reduction in the amplitude of the variability with
decreasing inclination, but also the change in the shape of the
light curve. This is particularly visible in a comparison of the
top and bottom panels of Fig. 7. In Fig. 8 we show the change
in R30 averaged over cycle 23 when we place the observer out of
the ecliptic plane. To facilitate comparison, we normalised each
value of the cycle-averaged variability, ⟨R30⟩i, to the correspond-
ing value for the ecliptic view, ⟨R30⟩90. Figure 8 shows that the
rotational variability decreases monotonically with decreasing 3.1. Approach We show the calculated solar light curve as it would be
observed by Kepler at various inclinations in Fig. 7. For this,
Table 2. Cycle-averaged R30 in mmag. 21
22
23
24
Mean
TSI
0.743
0.806
0.682
0.492
0.681
Kepler
0.808
0.872
0.731
0.530
0.735
CoRoT
0.801
0.866
0.726
0.526
0.730
TESS
0.680
0.739
0.615
0.445
0.620
Gaia GBP
0.944
1.020
0.861
0.625
0.862
Gaia G
0.817
0.883
0.741
0.537
0.744
Gaia GRP
0.684
0.742
0.617
0.447
0.623
VIRGO/blue
1.252
1.352
1.167
0.846
1.154
VIRGO/green
0.983
1.056
0.894
0.653
0.897
VIRGO/red
0.665
0.722
0.600
0.435
0.606
we divided the time series for cycle 23 into 90-day segments,
which correspond to Kepler quarters. Within each quarter, we
de-trended the light curves. 90◦corresponds to an ecliptic-bound
observer, 57◦represents a weighted mean value of the inclina-
tion with weights equal to the probability of observing a given
inclination (sin(i)) for the inclination i), and 0◦corresponds to an
observer facing the north pole. We additionally show 30◦as an
intermediate point between 57◦and 0◦. For the inclination values
of 30◦, 57◦, and 90◦, the variability is brought about by the solar
rotation, as well as the emergence and evolution of magnetic fea-
tures. A polar-bound observer does not observe the rotational
modulation because there is no transit of magnetic features and
the variability is merely generated by their emergence and evo-
lution (see Nèmec et al. 2020, for further details). We empha-
sise just the reduction in the amplitude of the variability with
decreasing inclination, but also the change in the shape of the
light curve. This is particularly visible in a comparison of the
top and bottom panels of Fig. 7. In Fig. 8 we show the change
in R30 averaged over cycle 23 when we place the observer out of
the ecliptic plane. To facilitate comparison, we normalised each
value of the cycle-averaged variability, ⟨R30⟩i, to the correspond-
ing value for the ecliptic view, ⟨R30⟩90. Figure 8 shows that the
rotational variability decreases monotonically with decreasing Table 2. Cycle-averaged R30 in mmag. Table 2. Cycle-averaged R30 in mmag. represent statistical properties of the Royal Greenwich Obser-
vatory sunspot record. We additionally randomised the longi-
tudes of the active-region emergences in the Jiang et al. (2011)
records. 3.1. Approach The SFTM is an advective-diffusive model for the passive
transport of the radial magnetic field on the surface of a star,
under the effects of large-scale surface flows. In this model, mag-
netic flux emerges on the stellar surface in the form of bipolar
magnetic regions (BMRs). We employed the SFTM in the form
given by Cameron et al. (2010) and followed the approach of
N20 to simulate light curves of the Sun at different inclinations
and with various filter systems. The emergence times, positions,
and sizes of active regions in our calculations were deter-
mined using the semi-empirical sunspot-group record produced
by Jiang et al. (2011). This synthetic record was constructed to The results presented in Sect. 2.3 are for the Sun viewed from the
ecliptic plane and apply to stars that are viewed approximately
equator-on. However, this is not always the case, and the incli-
nation of a star is often unknown. Calculations of the solar vari-
ability as it would be measured by an out-of-ecliptic observer
demand information about the distribution of magnetic features
on the far side (for the Earth-bound observer) of the Sun. N20
have used a surface flux transport model (SFTM) to obtain the
distribution of magnetic features of the solar surface, which was N.-E. Nèmec et al.: Connecting solar and stellar variabilities N.-E. Nèmec et al.: Connecting solar and stellar variabilities 0
1000
2000
3000
4000
5000
Kepler
s = 1.124
CoRoT
s = 1.110
TESS
s = 0.939
0
1000
2000
3000
4000
5000
R30 Filter [ppm]
GAIA GBP
s = 1.304
GAIA G
s = 1.131
GAIA GRP
s = 0.944
0
1000
2000
3000
R30 TSI [ppm]
0
1000
2000
3000
4000
5000
VIRGO/blue
s = 1.690
0
1000
2000
3000
R30 TSI [ppm]
VIRGO/green
s = 1.370
0
1000
2000
3000
R30 TSI [ppm]
VIRGO/red
s = 0.912
Fig. 4. Linear regression between the R30 values calculated with the TSI and with the solar light curves as they would be recorded in different filter
systems. The values of the slope, s, are given in the legend. The black lines have a slope equal to 1. Fig. 4. Linear regression between the R30 values calculated with the TSI and with the solar light curves as they would be recorded in different filter
systems. The values of the slope, s, are given in the legend. 3.1. Approach The black lines have a slope equal to 1. Fig. 4. Linear regression between the R30 values calculated with the TSI and with the solar light curves as they would b
systems. The values of the slope, s, are given in the legend. The black lines have a slope equal to 1. Table 1. Slopes of the linear regressions in Fig. 4. Table 2. Cycle-averaged R30 in mmag. 21
22
23
24
Mean
TSI
0.743
0.806
0.682
0.492
0.681
Kepler
0.808
0.872
0.731
0.530
0.735
CoRoT
0.801
0.866
0.726
0.526
0.730
TESS
0.680
0.739
0.615
0.445
0.620
Gaia GBP
0.944
1.020
0.861
0.625
0.862
Gaia G
0.817
0.883
0.741
0.537
0.744
Gaia GRP
0.684
0.742
0.617
0.447
0.623
VIRGO/blue
1.252
1.352
1.167
0.846
1.154
VIRGO/green
0.983
1.056
0.894
0.653
0.897
VIRGO/red
0.665
0.722
0.600
0.435
0.606 Table 1. Slopes of the linear regressions in Fig. 4. Slope
Kepler
1.123 (±0.007)
CoRoT
1.110 (±0.006)
TESS
0.939 (±0.004)
Gaia GBP
1.304 (±0.005)
Gaia G
1.131 (±0.005)
Gaia GRP
0.944 (±0.004)
VIRGO/blue
1.689 (±0.003)
VIRGO/green
1.370 (±0.007)
VIRGO/red
0.912 (±0.003)
represent statistical properties of the Royal Greenwich Obser-
vatory sunspot record. We additionally randomised the longi-
tudes of the active-region emergences in the Jiang et al. (2011)
records. Such a randomisation is needed to ensure that the near
and far side of the Sun have on average equal activity, which is
a necessary condition for reliable calculations of the inclination
effect. As a result, our calculations reproduce the statistical prop-
erties of a given solar cycle, but they do not represent the actual
observed BMR emergences for that specific cycle. We stress that
in N20 we developed the model outlined above to study the effect
of the inclination on the power spectra of solar brightness varia-
tions. Here we use this model to explicitly study the dependence
of the variability amplitude on the rotational timescale and its
dependence on the inclination in different filters. 3.2. Results
In the following, we place the observer out of the solar equa-
tor towards the solar north pole. This corresponds to inclinations
below 90◦. We quantify the rotational variability using the R30
metric introduced in the previous section. To represent an aver-
age level of solar activity, we limit the analysis to cycle 23, which
was a cycle of moderate strength. 3.2. Results In the following, we place the observer out of the solar equa-
tor towards the solar north pole. This corresponds to inclinations
below 90◦. We quantify the rotational variability using the R30
metric introduced in the previous section. To represent an aver-
age level of solar activity, we limit the analysis to cycle 23, which
was a cycle of moderate strength. We show the calculated solar light curve as it would be
observed by Kepler at various inclinations in Fig. 7. For this, A56, page 5 of 8 A56, page 5 of 8 A&A 638, A56 (2020) 0
200
400
600
800
1000
Time [days]
0
500
1000
1500
2000
2500
3000
R30 [ppm]
VIRGO/g+r
Kepler
Fig. 5. R30 over cycle 22 for VIRGO/g+r and Kepler. 0
200
400
600
800
1000
Time [days]
0
500
1000
1500
2000
2500
3000
R30 [ppm]
VIRGO/g+r
Kepler
Fig. 5. R30 over cycle 22 for VIRGO/g+r and Kepler. 0
500
1000 1500 2000 2500 3000
R30 VIRGO/g+r [ppm]
0
500
1000
1500
2000
2500
3000
R30 Kepler [ppm]
Kepler
s = 1.008
Fig. 6. Linear regression between the R30 calculated for VIRGO/g+r and
for Kepler light curves. The black solid line represents a linear regres-
sion with slope =1. the slopes of the linear regressions between the R30 values in dif-
ferent passbands and the TSI have to be corrected for the incli-
nation. When stellar measurements in the Kepler passband are
compared with the TSI records, this means the following: when
stars are observed from their equatorial planes, the variability in
Kepler is about 12% higher than in the TSI (see the slope given
in Table 1). However, when the Sun is compared to a group
of stars with random orientations of rotation axes, the inclina-
tion effect must be taken into account as well. It will reduce the
stellar variability observed in the Kepler passband by approx-
imately 15%. Coincidentally, these two effects almost exactly
cancel each other, and the observed TSI variability appears to
be a very good metric for the solar-stellar comparison of Kepler
stars in a statistical sense. We have shown in Sect. 2 that the amplitude of solar rota-
tional variability as it would be measured by Kepler can be
very accurately approximated by calculating the amplitude of the
VIRGO/g+r light curve. 3.2. Results However, when brightness variations of
the Sun are compared to those of a large group of stars with
unknown inclinations, the use solar variability averaged over all
possible inclinations should be used rather than the solar vari-
ability observed from the ecliptic plane. We have established
that the effect of a random inclination decreases the variability
in the Kepler passband by 15%. Taking this into account, the rel-
ative difference between the variability in VIRGO/g+r and the
solar variability in Kepler averaged over inclinations is −14%. Unlike for the TSI, corrections for the passband and the inclina-
tion only partly compensate for each other. We therefore suggest
that the TSI is a better representative of the Sun as it would be
observed by Kepler than VIRGO/g+r if the inclination of a star
is unknown. Fig. 5. R30 over cycle 22 for VIRGO/g+r and Kepler. Fig. 5. R30 over cycle 22 for VIRGO/g+r and Kepler. 0
500
1000 1500 2000 2500 3000
R30 VIRGO/g+r [ppm]
0
500
1000
1500
2000
2500
3000
R30 Kepler [ppm]
Kepler
s = 1.008 A56, page 6 of 8 4. Modelling Kepler light curves using VIRGO/SPM Fig. 6. Linear regression between the R30 calculated for VIRGO/g+r and
for Kepler light curves. The black solid line represents a linear regres-
sion with slope =1. The previous sections, we have quantitatively validated the argu-
ment of Basri et al. (2010) that the VIRGO/g+r light curve cor-
responds to the same variability as the Kepler light curve if both
light curves are recorded from the solar equatorial plane. In this
section we perform complementary calculations: we test if the
Kepler light curve can be modelled as a linear combination of
solar light curves in the different VIRGO/SPM channels. We
restrict our calculations to solar cycle 23. All light curves are
computed with the N20 model. inclination. This trend is seen across all considered filter sys-
tems. The differences in the inclination effect among the filter
systems are due to the different dependencies of the facular and
spot contrasts (as well as to their centre-to-limb variations) on
the wavelengths. p
We divided all light curves into 90-day segments and calcu-
lated the relative flux within these segments (i.e. we considered
the same normalisation of light curves as shown in Figs. 2 and 7). Next, we applied multiple linear regression to fit the Kepler light
curve with the VIRGO/SPM green+red light curves to determine
the best set of coefficients for the linear fit. g
To evaluate the averaged effect of the inclination, we intro-
duce a new measure that we call ⟨R30⟩and define as ⟨R30⟩=
P
i⟨R30⟩i · sin(i)
P
i sin(i)
,
(2) ⟨R30⟩=
P
i⟨R30⟩i · sin(i)
P
i sin(i)
, (2) For an ecliptic-bound observer, we write the multiple linear
regression in the form where i is the inclination. The factor sin(i) ensures that the cor-
responding values of ⟨R30⟩i are weighted according to the prob-
ability that a star is observed at inclination i. The ⟨R30⟩value
represents the variability of the Sun averaged over all possible
inclinations. In other words, if we observed many stars analo-
gous to the Sun with random orientations of the rotation axes,
their mean variability would be given by the ⟨R30⟩value. There-
fore ⟨R30⟩should be used for the solar-stellar comparison rather
than the ⟨R30⟩90 value. 4. Modelling Kepler light curves using VIRGO/SPM Synthetic solar light curves covering solar cycle 23 in the Kepler passband as it would appear at different in 90
80
70
60
50
40
30
20
10
0
Inclination [degree]
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
1.1
⟨R30⟩i ⟨ ⟨R30⟩90
Kep er
⟩oRoT
TESS
90
80
70
60
50
40
30
20
10
0
Inc ination [degree]
Gaia GBP
Gaia G
Gaia GRP
90
80
70
60
50
40
30
20
10
0
Inc ination [degree]
VIRGO⟨b ue
VIRGO⟨green
VIRGO⟨red
TSI
Fig. 8. Dependence of the mean variability in R30 on the inclination (termed ⟨R30⟩i, where i stands for the inclination). All values have be
normalised to the respective equator-on (i = 90◦) value, here called ⟨R30⟩90. Individual curves represent different filter systems. Left panel: Keple
TESS, and CoRoT. Middle panel: three Gaia filters. Right panel: three VIRGO filters and the TSI. The vertical dashed black line indicates
inclination of 57◦. 90
80
70
60
50
40
30
20
10
0
Inclination [degree]
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
1.1
⟨R30⟩i ⟨ ⟨R30⟩90
Kep er
⟩oRoT
TESS
90
80
70
60
50
40
30
20
10
0
Inc ination [degree]
Gaia GBP
Gaia G
Gaia GRP
90
80
70
60
50
40
30
20
10
0
Inc ination [degree]
VIRGO⟨b ue
VIRGO⟨green
VIRGO⟨red
TSI
Fig. 8. Dependence of the mean variability in R30 on the inclination (termed ⟨R30⟩i, where i stands for the inclination). All values have been
normalised to the respective equator-on (i = 90◦) value, here called ⟨R30⟩90. Individual curves represent different filter systems. Left panel: Kepler,
TESS, and CoRoT. Middle panel: three Gaia filters. Right panel: three VIRGO filters and the TSI. The vertical dashed black line indicates an
inclination of 57◦. Fig. 8. Dependence of the mean variability in R30 on the inclination (termed ⟨R30⟩i, where i stands for the inclination). All values have been
normalised to the respective equator-on (i = 90◦) value, here called ⟨R30⟩90. Individual curves represent different filter systems. Left panel: Kepler,
TESS, and CoRoT. Middle panel: three Gaia filters. Right panel: three VIRGO filters and the TSI. The vertical dashed black line indicates an
inclination of 57◦. Table 3. ⟨R30⟩/⟨R30⟩90 and ⟨R30⟩values for different filter systems. Table 3. ⟨R30⟩/⟨R30⟩90 and ⟨R30⟩values for different filter systems. 4. Modelling Kepler light curves using VIRGO/SPM (3) Ki, SPM = a · Vg + b · Vr,
(3) Ki, SPM = a · Vg + b · Vr, where i is the inclination, Vg and Vr are solar light curves in
VIRGO/SPM blue, green, and red filters (corresponding to the
equatorial plane). The best fit for i = 90◦yields a = 0.275
(±0.001) and b = 0.619 (±0.002). The r2 value is 0.999. Such
a high correlation is expected because the VIRGO/SPM and
Kepler rotational variability are similar, as discussed in Sect. 2. Next, we applied the multiple-regression model to simulate the
out-of-ecliptic Kepler-like light curve using the light curves in
the SPM channels as input. Because an inclination of 57◦is often
used to represent the statistical mean of all possible inclination
values, we fit the Kepler light curve observed at i = 57◦with a where i is the inclination, Vg and Vr are solar light curves in
VIRGO/SPM blue, green, and red filters (corresponding to the
equatorial plane). The best fit for i = 90◦yields a = 0.275
(±0.001) and b = 0.619 (±0.002). The r2 value is 0.999. Such
a high correlation is expected because the VIRGO/SPM and
Kepler rotational variability are similar, as discussed in Sect. 2. Next, we applied the multiple-regression model to simulate the
out-of-ecliptic Kepler-like light curve using the light curves in
the SPM channels as input. Because an inclination of 57◦is often
used to represent the statistical mean of all possible inclination
values, we fit the Kepler light curve observed at i = 57◦with a We present ⟨R30⟩normalised to ⟨R30⟩90 for all considered fil-
ter systems as well as the ⟨R30⟩values themselves in Table 3. In
the second column of Table 3 we give the inclination-corrected
value of the mean rotational variability from Table 2 for easier
application of our results. On average, all filter systems show a
15% lower variability than the equatorial case. This implies that N.-E. Nèmec et al.: Connecting solar and stellar variabilities -2000
-1500
-1000
-500
0
500
1000
Inclination = 90∘
-2000
-1500
-1000
-500
0
500
1000
Relative flux [ppm]
Inclination = 57∘
-2000
-1500
-1000
-500
0
500
1000
Inclination = ∘0∘
0
500
1000
1500
2000
2500
∘000
∘500
4000
4500
Time [da s]
-2000
-1500
-1000
-500
0
500
1000
Inclination = 0∘
Fig. 7. 4. Modelling Kepler light curves using VIRGO/SPM Synthetic solar light curves covering solar cycle 23 in the Kepler passband as it would appear at different inclinations. 90
80
70
60
50
40
30
20
10
0
Inclination [degree]
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
1.1
⟨R30⟩i ⟨ ⟨R30⟩90
Kep er
⟩oRoT
TESS
90
80
70
60
50
40
30
20
10
0
Inc ination [degree]
Gaia GBP
Gaia G
Gaia GRP
90
80
70
60
50
40
30
20
10
0
Inc ination [degree]
VIRGO⟨b ue
VIRGO⟨green
VIRGO⟨red
TSI
Fig. 8. Dependence of the mean variability in R30 on the inclination (termed ⟨R30⟩i, where i stands for the inclination). All values have
normalised to the respective equator-on (i = 90◦) value, here called ⟨R30⟩90. Individual curves represent different filter systems. Left panel: Ke
TESS, and CoRoT. Middle panel: three Gaia filters. Right panel: three VIRGO filters and the TSI. The vertical dashed black line indicat
inclination of 57◦. Table 3. ⟨R30⟩/⟨R30⟩90 and ⟨R30⟩values for different filter systems. linear combination of VIRGO light curves (observed at i = 9 -2000
-1500
-1000
-500
0
500
1000
Inclination = 90∘
-2000
-1500
-1000
-500
0
500
1000
Relative flux [ppm]
Inclination = 57∘
-2000
-1500
-1000
-500
0
500
1000
Inclination = ∘0∘
0
500
1000
1500
2000
2500
∘000
∘500
4000
4500
Time [da s]
-2000
-1500
-1000
-500
0
500
1000
Inclination = 0∘
Fig. 7. Synthetic solar light curves covering solar cycle 23 in the Kepler passband as it would appear at different inclinations. -2000
-1500
-1000
-500
0
500
1000
Inclination = 90∘
-2000
-1500
-1000
-500
0
500
1000
Relative flux [ppm]
Inclination = 57∘
-2000
-1500
-1000
-500
0
500
1000
Inclination = ∘0∘
0
500
1000
1500
2000
2500
∘000
∘500
4000
4500
Time [da s]
-2000
-1500
-1000
-500
0
500
1000
Inclination = 0∘
Fig. 7. Synthetic solar light curves covering solar cycle 23 in the Kepler passband as it would appear at different inclinations. Fig. 7. Acknowledgements.
We thank Chi-Ju Wu, whose master thesis has sparked the
idea for the manuscript. The research leading to this paper has received fund-
ing from the European Research Council under the European Union’s Hori-
zon 2020 research and innovation program (Grant agreement No. 715947). 5. Conclusions SKS acknowledges financial support from the BK21 plus program through
the National Research Foundation (NRF) funded by the Ministry of Educa-
tion of Korea. YCU acknowledges support through STFC consolidated grants
ST/N000838/1 and ST/S000372/1. We presented recipes for treating two problems that hamper the
comparison between solar and stellar rotational brightness vari-
ations: the difference between the spectral passbands that are
used for solar and stellar observations, and the effect of incli-
nation. To quantify the effect of different spectral passbands on
the rotational variability represented through the R30 metric, we
employed the SATIRE-S model. We found that the rotational
variability observed through the filter systems used by the Kepler
and CoRoT missions is about 12% higher than the TSI, whereas
the variability in the TESS passband is about 7% lower. For
Gaia G, we find +15% and for Gaia GBP +30% difference in
the amplitude of the rotational variability compared to the TSI,
whereas Gaia GRP shows a difference of −7%. These numbers
are valid for equator-on observations on rotational timescales. 4. Modelling Kepler light curves using VIRGO/SPM ⟨R30⟩/⟨R30⟩90 [%]
⟨R30⟩[mmag]
TSI
85.1
0.58
Kepler
84.8
0.62
CoRoT
83.8
0.61
TESS
84.0
0.56
Gaia GBP
83.7
0.72
Gaia G
84.1
0.62
Gaia GRP
84.9
0.52
VIRGO/blue
83.9
0.98
VIRGO/green
85.3
0.76
VIRGO/red
83.4
0.50
Notes. For ⟨R30⟩we multiplied ⟨R30⟩/⟨R30⟩90 by the corresponding
value for ⟨R30⟩90 from Table 2. Time-averaging is performed over solar
cycle 23. linear combination of VIRGO light curves (observed at i = 90◦). The best fit results in the following coefficients: a = −0.421
(±0.018) and b = 1.418 (±0.027), with r2 = 0.83. Figure 9 compares the Kepler-like light curve with the
regression model using light curves in all three VIRGO fil-
ters for i = 90◦and i = 57◦. For the 90◦inclination case
(left panel in Fig. 9), the differences between the two light
curves are basically invisible, but for the 57◦case, the dif-
ferences are quite pronounced. These differences have various
origins. In particular, the transits of magnetic features would
take different times for the ecliptic and out-of-ecliptic observer. Furthermore, with decreasing inclination, the facular contribu-
tion becomes stronger, while the spot contribution weakens (see
e.g. Shapiro et al. 2016), which changes the shape of the light
curve. It is therefore necessary to take the actual distribution of
magnetic features into account when light curves are modelled
that were observed at different inclination angles. Notes. For ⟨R30⟩we multiplied ⟨R30⟩/⟨R30⟩90 by the corresponding
value for ⟨R30⟩90 from Table 2. Time-averaging is performed over solar
cycle 23. A56, page 7 of 8 A56, page 7 of 8 A&A 638, A56 (2020) A&A 638, A56 (2020)
1260
1280
1300
1320
1340
1360
1380
1400
Time [days]
−1000
−750
−500
−250
0
250
500
750
1000
No malised flux [ppm]
Keple
VIRGO/g+
1260
1280
1300
1320
1340
1360
1380
1400
Time [days]
Keple
VIRGO/g+
Fig. 9. Comparison between the Kepler and the regressed VIRGO/green+red light curve. Left: i = 90◦, and right: i = 57◦. For the coefficients of the
fit, see the main text. n the Kepler and the regressed VIRGO/green+red light curve. Left: i = 90◦, and right: i = 57◦. For the coefficients of the Fig. 9. Comparison between the Kepler and the regressed VIRGO/green+red light curve. Left: i = 90◦, and right: i = 57◦. For the coefficients of the
fit, see the main text. References Baglin, A., Auvergne, M., Boisnard, L., et al. 2006, in 36th COSPAR Scientific
Assembly, COSPAR Meeting, 36, 3749 Ball, W. T., Unruh, Y. C., Krivova, N. A., et al. 2012, A&A, 541, A27 Ball, W. T., Krivova, N. A., Unruh, Y. C., Haigh, J. D., & Solanki, S. K. 2014, J. Atmos. Sci., 71, 4086 Basri, G., Walkowicz, L. M., Batalha, N., et al. 2010, ApJ, 713, L155
Basri, G., Walkowicz, L. M., & Reiners, A. 2013, ApJ, 769, 37 Bordé, P., Rouan, D., & Léger, A. 2003, A&A, 405, 1137 Borucki, W. J., Koch, D., Basri, G., et al. 2010, Science, 327, 977 ameron, R. H., Jiang, J., Schmitt, D., & Schüssler, M. 2010, ApJ q
Previous studies have used combinations of the red and
green VIRGO light curves for solar-stellar comparisons (see e.g. Basri et al. 2010; Gilliland et al. 2011; Harrison et al. 2012). We
used linear regressions of the rotational variability between the
two combined VIRGO/SPM passbands and the solar variability
as Kepler would observe it, to test the goodness of this compari-
son. We find that the variability in Kepler is 7% higher than that
of VIRGO/green+red. Moreover, we found that the sum of the
VIRGO green and red light curves very accurately represents the
solar light curve in the Kepler passband. This is only valid for the
Sun observed from the ecliptic, however. We showed that a linear
combination of VIRGO/SPM passbands cannot accurately repro-
duce the solar Kepler light curve observed out of ecliptic. Castelli, F., & Kurucz, R. L. 1994, A&A, 281, 817 Danilovic, S., Solanki, S. K., Livingston, W., Krivova, N., & Vince, I. 2016,
A&A, 587, A33 Ermolli, I., Matthes, K., Dudok de Wit, T., et al. 2013, Atmos. Chem. Phys., 13,
3945 Evans, D. W., Riello, M., De Angeli, F., et al. 2018, A&A, 616, A4
Fligge, M., Solanki, S. K., & Unruh, Y. C. 2000, A&A, 353, 380
Fontenla, J. M., Avrett, E. H., & Loeser, R. 1993, ApJ, 406, 319
Fröhlich, C. 2012, Surv. Geophys., 33, 453
Fröhlich, C., Romero, J., Roth, H., et al. 1995, Sol. Phys., 162, 101
Fröhlich, C., Andersen, B. N., Appourchaux, T., et al. 1997, Sol. Phys., 170, 1
Gaia Collaboration 2016, A&A, 595, A1
Gilliland, R. L., Chaplin, W. J., Dunham, E. W., et al. 2011, ApJS, 197, 6 Evans, D. W., Riello, M., De Angeli, F., et al. References 2018, A&A, 616, A4
Fligge, M., Solanki, S. K., & Unruh, Y. C. 2000, A&A, 353, 380
Fontenla, J. M., Avrett, E. H., & Loeser, R. 1993, ApJ, 406, 319
Fröhlich, C. 2012, Surv. Geophys., 33, 453 ,
,
,
,
,
Fröhlich, C. 2012, Surv. Geophys., 33, 453 Gaia Collaboration 2016, A&A, 595, A1 Gilliland, R. L., Chaplin, W. J., Dunham, E. W., et al. 2011, ApJS, 197, 6 Harrison, T. E., Coughlin, J. L., Ule, N. M., & López-Morales, M. 2012, AJ, 143,
4 Jiang, J., Cameron, R. H., Schmitt, D., & Schüssler, M. 2011, A&A, 528, A82
Kopp, G. 2016, J. Space Weather Space Clim., 6, A30 p
g
p
We have calculated the dependence of the rotational vari-
ability on inclination by following the approach in Nèmec et al. (2020). In this approach, an SFTM was used to simulate the
distribution of magnetic features on the surface of the Sun,
which was then used to compute the brightness variations with
SATIRE. We find that across all filter systems discussed in
this study, the rotational variability drops by about 15% when
it is averaged over all possible directions of the rotation axis. Because the Kepler rotational variability as observed from the
ecliptic plane is 12% higher than the TSI rotational variability,
we conclude that the TSI is the best proxy for the solar rotational
brightness variations if they were observed by Kepler when the
inclination effect is considered. pp
p
p
Krivova, N. A., Solanki, S. K., Fligge, M., & Unruh, Y. C. 2003, A&A, 399, L1 z, R. L. 1992, Rev. Mex. Astron. Astrofis., 23, 45 Maxted, P. F. L. 2018, A&A, 616, A39 Nèmec, N. E., Shapiro, A. I., Krivova, N. A., et al. 2020, A&A, 636, A43 Ricker, G. R., Winn, J. N., Vanderspek, R., et al. 2014, in Space Telescopes and
Instrumentation 2014: Optical, Infrared, and Millimeter Wave, Proc. SPIE,
9143 914320 Ricker, G. R., Winn, J. N., Vanderspek, R., et al. 2014, in Space Telescopes and
Instrumentation 2014: Optical Infrared and Millimeter Wave Proc SPIE Ricker, G. R., Winn, J. N., Vanderspek, R., et al. 2014, in Space Telescopes and Instrumentation 2014: Optical, Infrared, and Millimeter Wave, Proc. SPIE,
9143, 914320 Shapiro, A. I., Solanki, S. K., Krivova, N. A., Yeo, K. L., & Schmutz, W. K. 2016, A&A, 589, A46 2016, A&A, 589, A46
Solanki, S. K., Krivova, N. References A., & Haigh, J. D. 2013, ARA&A, 51, 311 Solanki, S. K., Krivova, N. A., & Haigh, J. D. 2013, ARA&A, 51, 311 Unruh, Y. C., Solanki, S. K., & Fligge, M. 1999, A&A, 345, 635 Vieira, L. E. A., Norton, A., Dudok de Wit, T., et al. 2012, Geophys. Res. Lett.,
39, L16104 Witzke, V., Shapiro, A. I., Solanki, S. K., Krivova, N. A., & Schmutz, W. 2018,
A&A, 619, A146 Witzke, V., Reinhold, T., Shapiro, A. I., Krivova, N. A., & Solanki, S. K. 2020,
A&A, 634, L9 Yeo, K. L., Krivova, N. A., Solanki, S. K., & Glassmeier, K. H. 2014, A&A, 570,
A85 A56, page 8 of 8
|
https://openalex.org/W2008154911
|
https://ieeexplore.ieee.org/ielx7/84/7283693/07046354.pdf
|
English
| null |
Development and Optimization of Durable Microelectrodes for Quantitative Electroanalysis in Molten Salt
|
Journal of microelectromechanical systems
| 2,015
|
cc-by
| 8,646
|
1346 1346 JOURNAL OF MICROELECTROMECHANICAL SYSTEMS, VOL. 24, NO. 5, OCTOBER 2015 Development and Optimization of Durable
Microelectrodes for Quantitative
Electroanalysis in Molten Salt
Ewen O. Blair, Damion K. Corrigan, Jonathan G. Terry, Senior Member, IEEE,
Andrew R. Mount, and Anthony J. Walton, Senior Member, IEEE Ewen O. Blair, Damion K. Corrigan, Jonathan G. Terry, Senior Member, IEEE,
Andrew R. Mount, and Anthony J. Walton, Senior Member, IEEE temperatures are typically between 360 and 500 °C [1] and
dissolved reactive species often produce a highly corrosive
medium. Abstract—Microfabricated
square
electrodes
with
finely
controlled highly reproducible dimensions have been developed
for
electrochemical
analysis
of
high-temperature
molten
salt (MS). These microelectrodes have been fabricated using
photolithographic techniques on silicon wafers and have been
designed for operation in lithium chloride/potassium chloride
eutectic salt at and ∼500 °C. The electrodes are constructed from
a series of patterned layers, and their development has involved
a systematic study and optimization of a number of different
material combinations. This has resulted in a process for making
electrodes that represents a step change in capability, delivering
the first robust microelectrode device capable of quantitative
electroanalysis in a MS system at 500 °C. [2014-0273] Macroelectrodes are the current electrode of choice for
measurement of redox species in MSs. However, reproducible
quantitative measurements are difficult to perform because
physical properties such as wetting are not well understood
and the active area of the electrode can be difficult to
determine [7], [8]. The glasses employed in insulating such
electrodes are also subject to failure due to thermal stress
and corrosion thus changing the active electrode area over the
course of measurement [9]. Index Terms—Microelectrodes, molten salt, microfabrication,
high temperature. A number of studies report the development of electrode
systems for measurement in MS. One notable study by
Malinowska et al., employed gold disc electrodes capable of
operating at 650 °C in molten carbonate salt. This involved
device construction using laser fabrication techniques, to pro-
duce electrodes with radii between 200 µm and 1.6 mm [10]. Most crucially however these macroelectrodes still suffer
from
electroanalytical
disadvantages
including
being
heavily affected by solution convection, iR drop unduly
influencing the response and the relatively large electrode
surface
area
producing
an
unfavourable
signal-to-noise
ratio. This work is licensed under a Creative Commons Attribution 3.0 License. For more information, see http://creativecom er a Creative Commons Attribution 3.0 License. For more information, see http://creativecommons.org/licenses/by/3.0/ Color versions of one or more of the figures in this paper are available
online at http://ieeexplore.ieee.org. Manuscript
received
September
4,
2014;
revised
January
7,
2015;
accepted
January
28, 2015. Date of publication
February
19, 2015;
date of current version September 29, 2015. This work was supported
in part by the U.K. Engineering and Physical Sciences Research Council
through the REFINE Project under Grant EP/J000779/1, in part by the
European Commission through the FP7 EURATOM Project ACSEPT
under Grant 211267, and in part by the SMART Microsystems Programme
FS/01/02/10 IeMRC Flagship. Subject Editor P. M. Sarro. Manuscript
received
September
4,
2014;
revised
January
7,
2015;
accepted
January
28, 2015. Date of publication
February
19, 2015;
date of current version September 29, 2015. This work was supported
in part by the U.K. Engineering and Physical Sciences Research Council
through the REFINE Project under Grant EP/J000779/1, in part by the
European Commission through the FP7 EURATOM Project ACSEPT
under Grant 211267, and in part by the SMART Microsystems Programme
FS/01/02/10 IeMRC Flagship. Subject Editor P. M. Sarro.
E. O. Blair, J. G. Terry, and A. J. Walton are with the Scottish
Microelectronics Centre, Institute for Integrated Micro and Nano Systems,
School of Engineering, University of Edinburgh, Edinburgh EH8 9YL, U.K.
(e-mail: e.blair@ed.ac.uk; jon.terry@ed.ac.uk; anthony.walton@ed.ac.uk).
D.
K.
Corrigan
and
A.
R.
Mount
are
with
the
Edinburgh
and
St.
Andrews
Research
School
of
Chemistry,
School
of
Chemistry,
University
of
Edinburgh,
Edinburgh
EH8
9YL,
U.K.
(e-mail:
dcorriga@staffmail.ed.ac.uk; a.mount@ed.ac.uk).
C l
i
f
f h
fi
i
hi
il bl g
p
j
E. O. Blair, J. G. Terry, and A. J. Walton are with the Scottish
Microelectronics Centre, Institute for Integrated Micro and Nano Systems,
School of Engineering, University of Edinburgh, Edinburgh EH8 9YL, U.K.
(e-mail: e.blair@ed.ac.uk; jon.terry@ed.ac.uk; anthony.walton@ed.ac.uk). This work is licensed under a Creative Commons Attribution 3.0 License. For more information, see http://creativecommons.org/licenses/by/3.0/ I. INTRODUCTION T
HE USE of a molten salt (MS) as an electrolytic medium
delivers a number of advantages including a large poten-
tial window, high ionic (and therefore electrical) conductivity,
and fast reaction kinetics [1]. These benefits facilitate the
production, stabilisation, and analysis of species that would
normally react with water and as a result MS has received
significant attention in the areas of metal manufacturing,
renewable energy, and nuclear reprocessing [2]–[5]. Chloride
melts are a particularly attractive system for such applications
as they have a relatively low melting point and are less corro-
sive than fluoride melts [6]. In spite of its favourable properties
when compared to fluoride melts, LiCl-KCl eutectic (LKE) is
still a challenging system in which to work. The operating T Microelectrodes (electrodes with a critical dimension in
the tens of micrometres range) exhibit superior electroanalyt-
ical properties when compared to macroelectrodes [11], [12]. These include higher signal-to-noise ratio, faster response
times, lower susceptibility to convection in the electrolyte, and
the ability to rapidly reach a steady-state current [13], [14]. Normally, high temperature electrochemistry refers to
studies carried out between 70 and 250 °C [15] and high
temperature microelectrodes are usually limited to operating
temperatures between 70 and 300 °C [16]–[18]. Traditional
microelectrodes, where a wire is encapsulated in glass, have
also been fabricated for measurements in MSs. However,
these electrodes are prone to chemical attack and thermal
degradation [19], as well as being difficult to make with
reproducible dimensions. (
j
y
y
)
D. K. Corrigan
and
A. R. Mount
are
with
the
Edinburgh
and
St. Andrews
Research
School
of
Chemistry,
School
of
Chemistry,
University
of
Edinburgh,
Edinburgh
EH8
9YL,
U.K. (e-mail:
dcorriga@staffmail.ed.ac.uk; a.mount@ed.ac.uk). Clearly microelectrodes that can function reliably in a
MS have the potential to deliver accurate, quantitative analysis
of the chemical species present in the salt and provide an
invaluable
sensor
for
a
range
of
industrial/process Color versions of one or more of the figures in this paper are available
online at http://ieeexplore.ieee.org. Color versions of one or more of the figures in this paper are available
online at http://ieeexplore.ieee.org. Digital Object Identifier 10.1109/JMEMS.2015.2399106 Digital Object Identifier 10.1109/JMEMS.2015.2399106 BLAIR et al.: DEVELOPMENT AND OPTIMIZATION OF DURABLE MICROELECTRODES 1347 Fig. 1. (a) A schematic layout of the device. (b) A transverse cross-section
of the device through the contact pad showing the layers of the device. I. INTRODUCTION The thickness of the layers has been exaggerated for clarity. sensing
systems,
including
nuclear
fuel
reprocessing,
and electroplating. The layer by layer nature of microfabrication presents
an opportunity to systematically optimise microelectronic
architectures through identification and understanding of
failure
mechanisms. Through
this
methodology
devices
have been designed that can operate reliably in the harsh
environment of LKE. When designing microelectrodes for
operation in LKE, it is necessary to consider that both
chemical
attack
and
electrochemical product
generation
may impact upon performance. For example, the lithium
ion is very small and able to permeate, destabilise, and
intercalate into a wide range of glass, ceramic, and crystalline
materials [20], [21]. Therefore effective barrier materials are
required. Thermal and intrinsic stress must also be managed in
order to prevent cracking or delamination at high temperatures. p
g
g
p
Reference [22] reports our initial study which identified the
potential of microfabricated electrodes for operation in high
temperature, corrosive environments such as LKE. Photolitho-
graphic techniques were chosen to fabricate MS compatible
microelectrodes because they enable the manufacture of large
numbers of electrodes with precise reproducible control over
their geometries and positions. However, these electrodes were
found to suffer from short operating lifetimes, a limited poten-
tial window of operation, and increased electrode area as they
degraded, making it impossible to extract quantitative informa-
tion due to uncertainty over the electrode area. The technology
is further developed in this paper, taking full advantage of
processes used in the fabrication of silicon integrated circuits,
where the required patterns can be repeatedly defined at the
sub-micrometre scale using photolithography [23]. Fig. 1. (a) A schematic layout of the device. (b) A transverse cross-section
of the device through the contact pad showing the layers of the device. The thickness of the layers has been exaggerated for clarity. 3. Maintain
its
overall
integrity
across
the
required
potential window and over the temperature range
studied; This paper first reports the design and fabrication of a
benchmark electrode capable of surviving in LKE for short
periods of time (∼5 mins). The failure mechanisms of
the device are then identified and methods of overcoming
them are described. The result is a process capable of
producing devices that make possible accurate, reliable
electrochemical measurements in LKE for over 30 minutes
without
any
performance
degradation. I. INTRODUCTION In
addition,
the
electrochemical response of the device in the presence of
the model redox agent silver (I) chloride is reported as part
of the procedure to confirm the successful construction of
a fully functional microelectrode with high dimensional
control. 4. Display quantitative and reproducible behaviour in its
electrochemical response that typifies a microelectrode,
ideally predicted by theory and corroborated by previous
studies. For this work, a range from −1.5 V to +0.5 V was selected
as the potential window for operation as this allows the
electrochemical detection of Uranium and Americium (which
are two important species found in spent nuclear fuel) along
with the detection of many industrially important metals such
as Zinc and Aluminium [25]–[27]. II. OBJECTIVES This paper defines microelectrodes as electrodes where at
least one critical dimension is in the tens of micrometres
range (the term ultramicroelectrode is used for electrodes
in the single micrometre range) [24]. To be considered a
functioning microelectrode of high fidelity, the device must
have the following characteristics: Figure 1 presents (a) the layout and (b) the cross-section
of the device architecture. These show how the electrode
and the contact pad dimensions are defined by the openings
etched through the top insulator. The large (6 by 4 mm)
contact pad was designed to enable simple, reliable connection
using a crocodile clip. The separation between contact pad
and electrode was designed to ensure that when the electrode
was immersed in LKE, the solution did not reach the contact
pad through wetting. In this work microsquare electrodes
with the range of edge lengths (L) 10 µm, 20 µm, 30 µm,
50 µm, and 100 µm were studied. 1. Be chemically inert in the melt thereby minimising
susceptibility to chemical attack; 2. Have an effective top insulation layer which defines the
electrode area and is able to operate as such within the
required range of the applied potential; JOURNAL OF MICROELECTROMECHANICAL SYSTEMS, VOL. 24, NO. 5, OCTOBER 2015 1348 Fig. 2. Cross-sections of the fabrication procedure used to produce the
microelectrodes. (a) An underlying insulator of SiO2 or Si-rich SiN is
grown/deposited on a silicon wafer. (b)-(f) The tungsten electrode metal area
is then defined using a pattern and lift-off technique. (g) The top insulator
of Si-rich SiN or Si3N4 is then deposited. (h)-(i) The areas to become the
microelectrode and contact pad are defined using photolithography. (j) The
exposed areas are then etched to expose the electrode metal. (k) The resist is
then stripped and the device is completed. re-entrant profile in the remaining resist shown in figure 2(d). The electrode metallisation, which comprises of a 20 nm
thick metal adhesion layer (titanium or titanium nitride)
covered by a thicker film of the electrode metal (tungsten),
is then deposited by DC magnetron sputtering (figure 2(e)). The remaining resist is then removed, which lifts off the
unwanted metal and leaves behind the desired electrode
metallisation pattern shown in figure 2(f). A top 500 nm
thick dielectric (silicon-rich silicon nitride or stoichiometric
silicon nitride) is then deposited over the metal to insulate it
from the MS (figure 2(g)). Fig. 2. Cross-sections of the fabrication procedure used to produce the
microelectrodes. (a) An underlying insulator of SiO2 or Si-rich SiN is
grown/deposited on a silicon wafer. (b)-(f) The tungsten electrode metal area
is then defined using a pattern and lift-off technique. (g) The top insulator
of Si-rich SiN or Si3N4 is then deposited. (h)-(i) The areas to become the
microelectrode and contact pad are defined using photolithography. (j) The
exposed areas are then etched to expose the electrode metal. (k) The resist is
then stripped and the device is completed. II. OBJECTIVES Finally, a layer of positive resist
is spun on to the wafer, selectively exposed (figure 2(h))
and developed (figure 2(i)). The exposed top insulator is
then etched, to expose the metal electrodes and contact
pad (figure 2(j)). The remaining resist is then removed and
the completed device is ready for testing (figure 2(k)). V. EXPERIMENTAL Once fabricated, in preparation for characterisation in LKE
at 500 °C, crocodile clips were crimped to a tungsten wire
for the electrical connection to a potentiostat. The crocodile
clip and bond pad were then encapsulated in a heat-resistant
putty to provide both physical and chemical protection of
the connection. All the different variants of the device were
characterised in 100g of LKE (45g of LiCl and 55g of KCl) in
a vitreous carbon crucible located in a quartz cell heated in a
vertical tube furnace. The LKE was melted and maintained
under an argon atmosphere and cyclic voltammetry with
silver (I) chloride as the redox agent was then used to
determine the functionality of the devices. Silver chloride
was chosen because it displays characteristic electrochemical
plating and stripping behaviour on macroelectrodes in LKE
at moderate voltages. It is also a simple and stable redox
agent which is easily handled, making it an ideal compound
for initially characterising electrochemical performance of
the devices. An Ag/Ag+ reference electrode was formed by
sealing a silver wire with 1% by mass Ag+ in LKE in a mullite
tube. All potentials quoted in this paper are with respect to this
electrode. A 1.8 mm diameter tungsten wire was employed as
the counter electrode. A. Benchmark Device Multiple materials were characterised in this study to
identify the combinations and characteristics required for
electrode systems to successfully operate in the chemically
harsh environment of LKE melts at 500 °C. The electrodes
were fabricated on 100 mm diameter <100> p-type silicon
wafers and figure 2 shows the base fabrication process. This
process starts with a 500 nm insulation layer being grown
(silicon dioxide)/deposited (silicon nitride) on the wafer
(figure 2(a)), which electrically isolates the silicon substrate
from the electrode device. Next a layer of negative photoresist
is spin coated onto the wafer and baked (figure 2(b)). Figure 2(c) shows the photoresist being selectively exposed
to ultraviolet light. Subsequent development of the resist
results in removal of the unexposed material, leaving the The initial benchmark fabrication process for this work was
based upon the electrodes detailed in [22]. These comprised of
a 500 nm thick underlying insulation layer of LPCVD silicon-
rich silicon nitride (Si-rich SiN) to insulate the electrode metal
from the underlying silicon wafer. Silicon nitride was selected
because it is chemically inert and physically robust [28]. Stoichiometric silicon
nitride (Si3N4) has
a very high
intrinsic stress [29] and hence lower-stress Si-rich SiN had
been selected to minimise this. A 20 nm titanium layer
was used to provide adhesion [30] between the underlying
insulator and the electrode metal, which consisted of a 200 nm
tungsten film. Tungsten was employed because it is a common
macroelectrode material used for electrochemical measure-
ments in MS as it is electrochemically inert between the BLAIR et al.: DEVELOPMENT AND OPTIMIZATION OF DURABLE MICROELECTRODES 1349 Fig. 3. (a) Silver deposits on the surface of the top insulator of the benchmark
device after it was cycled between +0.5V and −0.5V for 10-15 minutes. (b) A microsquare electrode on a benchmark device, where a section of metal
has detached after cycling for 5-10 minutes between +0.5V and −0.5V. electrochemistry in LKE with these devices and presented the
following series of challenges: electrochemistry in LKE with these devices and presented the
following series of challenges: 1. Delamination of the top insulator; 1. Delamination of the top insulator; 2. Susceptibility of the electrode metal to detachment; 2. Susceptibility of the electrode meta 3. The top insulator not operating as an effective barrier to
electrochemistry at the underlying metal. B. Failure Analysis Whilst restricting
the operational voltage limits avoided delamination it indicated
that the top insulator was ineffective in preventing the reduc-
tion of silver ions at the underlying tungsten according to An example of a very successful stress relief strategy for
the high level of intrinsic stress present in silicon nitride is the
LOCOS process which is used for growing the field oxide in
CMOS technology [33]. As noted above, SiO2 has a compres-
sive stress and if this is matched with the tensile stress in the
Si-rich layer SiN, wafer bow can be eliminated [29], [34]. This
was consequently the approach used to relieve stress in the Si-
rich SiN top insulator and thereby reduce the probability of
delamination. Hence, devices were fabricated with a 500 nm
thermally grown SiO2 layer in place of the 500 nm Si-rich
SiN base insulation layer used in the benchmark device. It was
satisfying that when these devices were electrochemically
cycled in the melt the stress levels were sufficiently reduced
to the point where no delamination of the top SiN insulator
layer was observed. The extrinsic stress also needs to be considered and is
mainly related to the thermal expansion mismatch between
the deposited layer and the silicon substrate. In the electrodes
fabricated in this paper it originates from the strain resulting
from the wafer cooling to its room temperature dimensions
after deposition. This bi-axial thermal mismatch stress is
typically less than the intrinsic stress. Assuming the strain is
set by the much thicker silicon wafer it can be calculated to
be 84MPa for Si-rich SiN and 151MPa for SiO2 for the wafer
operating at 500 °C. Ag+
(solv) + e−→Ag(s)
(1) (1) Occasionally, in this restricted potential window, silver
plating/stripping currents were also seen to decrease. When the
device was removed from LKE and inspected, flakes of metal
were missing from the microsquare as shown in figure 3(b). This implied there was also either poor adhesion between the
layers or the electrode metal and/or the adhesion layer were
becoming exposed to LKE and subsequently attacked leading
to a reduction in the overall area of the tungsten. Figure 4(a) and (b) compares devices fabricated with Si-rich
SiN and SiO2 underlying insulators respectively after cycling
between −1.5 V and +0.5 V. B. Failure Analysis 1) Stress: The delaminations observed in the benchmark
device architecture indicated that excessive stress was being
generated in the layered structure. There are two sources of the
stress resulting from layer deposition; intrinsic (related to the
internal structure of the film resulting from its deposition) and
extrinsic (largely resulting from thermal-mismatch between
layers). Depending on process conditions typically the intrinsic
stress in Si-rich SiN is tensile and SiO2 compressive [29] with
measured magnitudes of 375 ± 40MPa and 272 ± 34MPa
respectively. These values agree with the literature [29] and
were obtained using profilometry and the Stoney formula [32]. solvent limits of LKE [31]. The top insulator which defined the
microelectrode and insulated the tungsten interconnect from
the MS solution was also a 500 nm layer of Si-rich SiN. Devices fabricated using this material combination were
characterised by initially submerging them in LKE for half
an hour at 500 °C after which they were removed and
examined under a microscope. This showed no obvious signs
of chemical attack (such as discolouration or surface damage). When potentials ranging between −1.5 V and +0.5 V were
applied, the devices operated successfully, passing the currents
associated with silver stripping (below −0.3V) and plating
(above −0.3V) in the nA range for one to ten minutes, after
which the currents increased markedly into the mA range. This
increase in current was indicative of a failure in the top insula-
tor leading to exposure of additional tungsten. Upon removal
from LKE and inspection, it was found in these cases that the
top insulation layer had delaminated, which was believed to
have been caused by stress induced through electrochemical
cycling. No delamination was observed when restricting the
potential window to between −0.5 V and +0.5 V but the
characteristic electrochemistry of silver plating and stripping
could still be observed. However, it was also noticed that even
in this reduced voltage window there were deposits of silver
on the areas of the top insulator overlying the metal, as shown
in figure 3(a), and the larger than expected currents persisted
despite the absence of delamination events. A. Benchmark Device In the following sections each of these failure mechanisms is
analysed systematically and a solution to each mode of failure
is presented. The end result is an optimised device capable of
operating in the LKE environment. In the following sections
it can be presumed unless stated otherwise that devices were
evaluated by electrochemically cycling them over the voltage
range −1.5 to +0.5 V. Fig. 3. (a) Silver deposits on the surface of the top insulator of the benchmark
device after it was cycled between +0.5V and −0.5V for 10-15 minutes. (b) A microsquare electrode on a benchmark device, where a section of metal
has detached after cycling for 5-10 minutes between +0.5V and −0.5V. B. Failure Analysis It can be observed the stress
relief provided by the underlying SiO2 successfully reduces the
overall stress and solves the delamination problem experienced
when Si-rich SiN is used as the underlying insulator. In
summary,
the
characterisation
of
the
benchmark
device highlighted the difficulties associated with performing JOURNAL OF MICROELECTROMECHANICAL SYSTEMS, VOL. 24, NO. 5, OCTOBER 2015 1350 Fig. 6. Device
fabricated
with
a
Si3N4
top
insulator
following
electrochemical cycling at 500 °C for half an hour (left) and a magnified
area of the surface showing no damage or silver deposition (right). Fig. 4. (a) Benchmark device with an Si-rich SiN underlying insulator
where the top insulator has delaminated. (b) Device fabricated with a SiO2
underlying insulator which shows no delamination of the top insulator. The
two devices were electrochemically cycled for 5 minutes at 500 °C. Fig. 6. Device
fabricated
with
a
Si3N4
top
insulator
following
electrochemical cycling at 500 °C for half an hour (left) and a magnified
area of the surface showing no damage or silver deposition (right). Fig. 4. (a) Benchmark device with an Si-rich SiN underlying insulator
where the top insulator has delaminated. (b) Device fabricated with a SiO2
underlying insulator which shows no delamination of the top insulator. The
two devices were electrochemically cycled for 5 minutes at 500 °C. For the seed layer to be responsible for the electrode film
removal, LKE must be able to reach the underlying titanium
adhesion layer. The sporadic nature of this effect suggests
it was most likely due to defects/pinholes in the electrode
metal film. As pinholes are difficult to completely remove,
it is advantageous to employ an adhesion layer which is not
electrochemically dissolved by LKE over the required poten-
tial range. Titanium nitride is known to offer good corrosion
resistance and is an often used barrier material [35], [36]. To investigate the chemical and electrochemical response of
deposited TiN in the salt, 20 nm of TiN was sputtered onto
500 nm of LPCVD Si-rich SiN and diced into strips. After
being submerged in the salt, the TiN showed no signs of
dissolution. The sample was then subjected to cyclic voltam-
metry for 15 minutes at a sweep rate of 200 mVs−1. The
titanium nitride was not electrochemically dissolved when
cycled between −1.5 V and +0.5 V. B. Failure Analysis Finally, to confirm the
improved resistance of a combined tungsten metal layer and
TiN adhesion layer to the salt, a device fabricated without
a top insulator but with a 20 nm TiN adhesion layer and
200 nm tungsten layer was cycled in the melt for 30 minutes
in the same potential window. The response was unchanged
with time indicating resistance of the adhesion layer to
electrochemical dissolution in LKE, as shown in figure 5(b). This confirmed that TiN was a suitable adhesion layer for these
devices. Fig. 5. (a) Electrode with a strip of tungsten on a titanium adhesion layer
where a large section of the titanium has been electrochemically stripped,
removing the overlying tungsten. (b) A tungsten electrode with a titanium
nitride adhesion layer which has been unaffected by electrochemical cycling. Fig. 5. (a) Electrode with a strip of tungsten on a titanium adhesion layer
where a large section of the titanium has been electrochemically stripped,
removing the overlying tungsten. (b) A tungsten electrode with a titanium
nitride adhesion layer which has been unaffected by electrochemical cycling. However, it is also well known that silicon dioxide is
chemically attacked by LKE. To confirm this, when a 500 nm
film of thermally grown silicon dioxide was immersed in LKE,
it was completely removed in under 10 minutes. Hence,
an important design consideration is that the silicon dioxide
underlying layer is either never directly exposed to LKE,
or its exposure is limited so as to not impact on device
lifetime. 3) Top Insulator: It was observed that the Si-rich SiN top
insulator was not particularly robust and often failed to insulate
successfully from the molten salt. It was possible that this
was due to the Si-rich SiN not acting as an impermeable
chemical barrier. It was expected that stoichiometric silicon
nitride (Si3N4) would provide a better barrier to LKE than
Si-rich SiN. This is because (a) Si-N bonds are more covalent
in character than Si-Si bonds, making them more resistant
to chemical attack, and (b) the material is denser than other
SixNy ratios [37]. 2) Adhesion Layer: To investigate the effect of metal
detachment from the microsquare, a simple device consisting
of a tungsten electrode metal film upon a titanium adhesion
layer was connected and electrochemically cycled. In this
case the electrochemical currents reduced to zero in under
five minutes indicating loss of electrode metal. B. Failure Analysis Figure 5(a)
shows a sample following removal from the melt visually
confirming this effect. As tungsten is a well-used electrode
metal in LKE [30], it seems unlikely this was the source of
the metal detachment. It was suspected that electrochemical
dissolution of the underlying adhesion layer was responsible. To identify whether this was the case, a titanium wire was
submerged into LKE and electrochemically cycled and was
found to electrochemically dissolve at a potential of ∼0 V. Figure 6 shows a device with a Si3N4 top insulator after
removal from the melt following half an hour of cycling. It can be observed that there is no visible degradation and the
top insulation layer of the device shows no silver deposited
on the surface. BLAIR et al.: DEVELOPMENT AND OPTIMIZATION OF DURABLE MICROELECTRODES 1351 Fig. 7. (a) CV of silver plating and stripping using benchmark electrode. (b) CV of silver plating and stripping using an optimised electrode. experimentally observed area was consistent with the Si-rich
SiN providing incomplete passivation as previously discussed
and illustrated in figure 3(a) and/or with delamination of the
top insulator as shown in figure 4(a). In contrast, figure 7(b) shows a cyclic voltammogram
from an optimised electrode with L = 20 µm. The most
immediate thing to note is that the current scale on figure 7(b)
is now in the order of nA as opposed to µA in figure 7(a),
which in itself is indicative of a microelectrode. Also evident
in figure 7(b) is the sharp stripping peak and limiting
current, arising from the diffusion-controlled mass transport,
characteristic of microelectrodes. Using equation (2), an edge
length of 19.6 µm was calculated at 450 °C for the optimised
electrode. This is a highly satisfying finding, as L is within
2% of the designed value, well within the tolerance reported
for high fidelity ambient microelectrode systems [40]. This
compares favourably with the electrodes in [10], where
the electrode radius was determined electrochemically to
be 40% larger than expected under ambient conditions. It should also be noted that the charge passed during silver
plating was the same (0.7 µC) as that passed during stripping. When this device was cycled using a range of scan rates, the
electrochemical response also proved to be independent of
scan rate further indicating that the device was performing
as expected for a 20 µm square microelectrode. B. Failure Analysis Finally,
the device was cycled in the melt for 30 minutes with no
change in the electrochemical response. Visual inspection
after
electrochemical
cycling
showed
an
unblemished
top insulator film with tungsten metal still present in
the
previously
defined
electrode
area. These
analyses
confirmed the successful production of working optimised
devices overcoming the limitations identified in [7]–[19]. A more comprehensive electrochemical characterisation of
these electrodes is presented in a companion publication [41]. Fig. 7. (a) CV of silver plating and stripping using benchmark electrode. (b) CV of silver plating and stripping using an optimised electrode. It should be noted that stoichiometric Si3N4
has a
larger intrinsic tensile stress, which was measured to be
950 ± 24MPa in agreement with literature values [29], [34]. Despite this increased tensile stress; the 500nm silicon dioxide
continued to provide adequate stress relief and no delamination
of the devices was observed. These devices also produced the
desired quantitative electrochemical response, as reported in
the next section. VII. ELECTRODE PERFORMANCE - ELECTROCHEMICAL
CHARACTERISATION OF SILVER (I) CHLORIDE IN LKE After fabrication of the electrodes they were quantitatively
characterised to identify fully functioning devices. To confirm
the fabricated microelectrode was of the correct dimensions,
the edge length was determined using the established expres-
sion for the limiting current for a square microelectrode, which
is given by [38] iL = 2.341nFDcL,
(2) (2) where iL is the limiting current, n is the number of electrons
transferred, F is Faraday’s constant, D is the diffusion coef-
ficient, c is the concentration of the redox agent and L is the
microsquare edge length. The extraction of the expected edge
length was considered to be a good indicator of a high-fidelity
electrode and was used below for both the original benchmark
device and the final optimised device, which incorporated the
SiO2 underlying insulator, the TiN adhesion layer, and the
Si3N4 top insulator. REFERENCES SUMMARY OF MATERIAL COMBINATIONS STUDIED FOR ANALYSIS IN MS SUMMARY OF MATERIAL COMBINATIONS STUDIED FOR ANALYSIS IN MS [1] J.-C. Poignet and J. Fouletier, “Physico-chemical properties of molten
salts,” in Materials Issues for Generation IV Systems, V. Ghetta,
D. Gorse, D. Mazière, V. Pontikis, Eds. Berlin, Germany: Springer-
Verlag, 2008, pp. 523–536. [2] D. J. Fray, “Emerging molten salt technologies for metals production,”
J. Minerals, Metals Mater. Soc., vol. 53, no. 10, pp. 27–31, Oct. 2001. [Online]. Available: http://link.springer.com/article/10.1007%2Fs11837-
001-0052-5 [3] B. Mishra and D. L. Olson, “Molten salt applications in materials
processing,” J. Phys. Chem. Solids, vol. 66, nos. 2–4, pp. 396–401, 2005. [4] M. Straka, L. Szatmáry, M. Marecek, and M. Korenko, “Uranium recov-
ery from LiF–CaF2–UF4–GdF3 system on Ni electrode,” J. Radioanal. Nucl. Chem., vol. 298, no. 1, pp. 393–397, Oct. 2013. [Online]. Available:
http://link.springer.com/article/10.1007%2Fs10967-013-
2436-8 [5] A. Gil
et
al.,
“State
of
the
art
on
high
temperature
thermal
energy storage for power generation. Part 1—Concepts, materials
and modellization,” Renew. Sustain. Energy Rev., vol. 14, no. 1,
pp. 31–55, Jan. 2010. [Online]. Available: http://www.sciencedirect.com/
science/article/pii/S1364032109001774 with the Si-rich SiN used in the previously reported process. Whilst the Si-rich SiN film had lower internal stress, when it
is employed as the top insulator the measured currents are
larger than expected as a result of incomplete passivation. This behaviour is not observed when a stoichiometric Si3N4
top insulator is employed. The dramatic improvement in
passivation between the two silicon nitride layers also suggests
it was not simply pinholes or defects in the top insulation layer,
but the superior chemical resistance of Si3N4. [6] I. Yasuhiko
and
N. Toshiyuki,
“Non-conventional
electrolytes
for
electrochemical
applications,”
Electrochim. Acta,
vol. 45,
nos. 15–16, pp. 2611–2622, May 2000. [Online]. Available: http://
www.sciencedirect.com/science/article/pii/S0013468600003418 [7] M. R. Bermejo,
J. Gómez,
J. Medina,
A. M. Martínez,
and
Y. Castrillejo, “The electrochemistry of gadolinium in the eutec-
tic LiCl–KCl on W and Al electrodes,” J. Electroanal. Chem.,
vol. 588,
no. 2,
pp. 253–266,
Mar. 2006. [Online]. Available:
http://www.sciencedirect.com/science/article/pii/S0022072806000350 p
The engineering yield of the reported optimised devices
is close to 100% and when left in the melt for two weeks
with no electrical activation the devices appeared completely
unaffected. As expected, when electrically activated, the device
lifetime is heavily affected by the size of the potential window
over which it was scanned and the temperature of the melt. REFERENCES A variation in electrode lifespan has been observed with
the overwhelming majority of devices surviving between
0.5 hours and 2.5 hours of electrochemical activation when
scanned between −1.5 V and +0.5 V. There have been a few
device failures within 10 minutes and these short lifetimes
are almost certainly due to latent defects. The failure rates
can be further reduced by lowering particulate levels and in
future devices tighter process controls are being implemented
in order to improve operational lifetimes. [8] P. Baumli and G. Kaptay, “Wettability of carbon surfaces by pure molten
alkali chlorides and their penetration into a porous graphite substrate,”
Mater. Sci. Eng., A, vol. 495, nos. 1–2, pp. 192–196, 2008. [Online]. Available: http://www.kaptay.hu/pub/kaptay-j117.pdf [9] S. Senderoff, “Electrode reactions in molten salts,” in Proc. Symp. Electrochem. Process.,
1967,
pp. 32–36. [Online]. Available:
https://web.anl.gov/PCS/acsfuel/preprint%20archive/11_1_MIAMI_04-
67.htm [10] B. Malinowska, M. Cassirl, and J. Devynck, “Design of a gold
ultramicroelectrode for voltammetric studies at high temperature in
glass-corrosive media (molten carbonate at 650 °C),” J. Electrochem. Soc., vol. 141, no. 8, pp. 2015–2017, Aug. 1994. [Online]. Available:
http://jes.ecsdl.org/content/141/8/2015.full.pdf [11] R. J. Forster and T. E. Keyes, “Behaviour of ultramicroelectrodes,”
in Handbook of Electrochemistry, C. G. Zoski, Ed. Amsterdam,
The Netherlands: Elsevier, 2007, pp. 155–171. [12] M. Fleischmann and S. Pons, “The behavior of microelectrodes,” Anal. Chem., vol. 59, no. 24, pp. 1391A–1399A, 1987. [13] D. Pletcher, “Why microelectrodes?” in Microelectrodes: Theory and
Applications, I. Montenegro, M. A. Queirós, and J. L. Daschbach, Eds. Norwell, MA, USA: Kluwer, 1991, pp. 3–15. JOURNAL OF MICROELECTROMECHANICAL SYSTEMS, VOL. 24, NO. 5, OCTOBER 2015 1352 TABLE I VIII. DISCUSSION Analysis of three key failure mechanisms provided an
insight into the operation of microfabricated microelectrodes
in the high temperature environment of LKE MS. This
approach guided development of optimised devices that
functioned in this environment for at least half an hour. It has been shown that intrinsic stress is the most important
stress related factor when operating in the LKE environment. In the role of underlying insulator; the Si-rich SiN layer con-
tributes less thermal stress than the SiO2. However the intrinsic
stress is much higher and results in device delamination. There
was a concern that dicing into individual chips exposed the
SiO2 insulator to LKE along the cut edges. However no
detrimental effect has been observed over the time courses
investigated to date. Figure 7(a) shows the cyclic voltammogram from a bench-
mark electrode. It is important to note the high magnitude
of the current and the disparity between charges passed
during plating and stripping (often 5-10 times more current
was passed in plating than stripping). The explanations for
these two phenomena were silver plating on the metal areas
underlying the top insulator and the subsequent isolation of
deposited silver in the top insulator upon stripping. Both
of these effects can be explained in terms of incomplete
passivation by the top insulator. Using the established literature
value of 2.42×10−5cm2s−1 for the diffusion coefficient (D) of
silver (I) chloride at 457 °C [39] in LKE and equation (2),
the edge length calculated for the benchmark electrode
in fig 7(a) was 6.6 mm. Such a large discrepancy between
the defined microelectrode edge length (L = 50 µm) and the The necessity of an electrochemically inert adhesion layer
implies that there is infiltration of salt through the electrode
metal, most likely via pinholes. For a robust device it is clear
that the total performance of the material layers used must be
considered, even if they are not in apparent direct contact with
the salt. The use of stoichiometric silicon nitride (Si3N4) as the
top insulator provides a superior dielectric barrier compared BLAIR et al.: DEVELOPMENT AND OPTIMIZATION OF DURABLE MICROELECTRODES His
first
post-doctoral position (funded by GSK and in collaboration with GSK Barnard
Castle Plant) was a continuation of the work undertaken during his Ph.D. [28] T. L. Chu, C. H. Lee, and G. A. Gruber, “The preparation and
properties of amorphous silicon nitride films,” J. Electrochem. Soc.,
vol. 114, no. 7, pp. 717–722, 1967. [Online]. Available: http://jes.ecsdl. org/content/114/7/717.abstract g
[29] D. Flandre, J. Laconte, and J.-P. Raskin, “Thin dielectric films
stress
extraction,”
in
Micromachined
Thin-Film
Sensors
for
SOI-CMOS
Co-Integration. Berlin,
Germany:
Springer-Verlag,
2006, ch. 2, pp. 47–103. [Online]. Available: http://link.springer. com/chapter/10.1007/0-387-28843-0_3 He was with the Electrochemistry Group, University of Southampton,
Southampton, U.K., where he was involved in a project using microstructured
electrodes for the optical detection and electrochemical
discrimination
of
DNA
sequences,
before
moving
to
Edinburgh. He
received
the
Ph.D. degree
in
bioanalytical
chemistry
from
Cranfield
University,
Bedford, U.K., which was funded by GSK, and was involved in the
development
of
sensor
systems
to
address
specific
purity
problems
associated
with
large-scale
pharmaceutical
manufacture. His
first
post-doctoral position (funded by GSK and in collaboration with GSK Barnard
Castle Plant) was a continuation of the work undertaken during his Ph.D. [30] M. J. Madou, “Pattern transfer with additive techniques,” in Fundamen-
tals of Microfabrication, 2nd ed. Boca Raton, FL, USA: CRC Press,
2002, ch. 3, p. 126. [31] M. Misra, K. S. Raja, and J. Ruppert, “Electrochemical
corro-
sion behavior of refractory metals in LiCl-Li2O molten salt,” ECS
Trans., vol. 33, no. 7, pp. 181–192, 2010. [Online]. Available:
http://ecst.ecsdl.org/content/33/7/181.abstract [32] M. Zecchino and T. Cunningham, Thin Film Stress Measurement
Using
Dektak
Stylus
Profilers. Plainview,
NY,
USA:
Veeco,
2004. [Online]. Available:
http:/www.rpi.edu/dept/cie/mncr/
documents/AN516_Dektak_Stress_Measure.pdf p
documents/AN516_Dektak_Stress_Measure.pdf Jonathan
G. Terry
(SM’08)
received
the
B.Eng. degree
in
electronics
engineering,
the
M.Sc. degree in microelectronic material and device
technology, and the Ph.D. degree in solid-state
electronics
from
the
University
of
Manchester
Institute of Science and Technology, Manchester,
U.K. He joined the Institute for Integrated Micro
and
Nanosystems,
University
of
Edinburgh,
Edinburgh, U.K., in 1999, as a Research Fellow. He is currently a Chancellor’s Fellow and Lecturer
with the University of Edinburgh, where his main
area of interest is in the development of More-than-Moore technologies, and
the integration of novel fabrication processes and materials with foundry
CMOS to create smart microsystems. Jonathan
G. Terry
(SM’08)
received
the
B.Eng. degree
in
electronics
engineering,
the
M.Sc. degree in microelectronic material and device
technology, and the Ph.D. BLAIR et al.: DEVELOPMENT AND OPTIMIZATION OF DURABLE MICROELECTRODES BLAIR et al.: DEVELOPMENT AND OPTIMIZATION OF DURABLE MICROELECTRODES [38] H. L. Woodvine, J. G. Terry, A. J. Walton, and A. R. Mount, “The
development and characterisation of square microfabricated electrode
systems,” Analyst, vol. 135, no. 5, pp. 1058–1065, May 2010. [Online]. Available: http://www.ncbi.nlm.nih.gov/pubmed/20419257 [19] R. T. Carlin and R. A. Osteryoung, “Deposition studies of lithium
and
bismuth
at
tungsten
microelectrodes
in
LiCl:KCl
eutectic,”
J. Electrochem. Soc., vol. 136, no. 5, pp. 1249–1255, Jan. 1989. [Online]. Available: http://jes.ecsdl.org/content/136/5/1249.abstract pp
ailable: http://jes.ecsdl.org/content/136/5/1249.abstract [20] H. Wang, N. J. Siambun, L. Yu, and G. Z. Chen, “A robust alumina
membrane reference electrode for high temperature molten salts,”
J. Electrochem. Soc.,
vol. 159,
no. 9,
pp. H740–H746,
2012. [Online]. Available:
http://jes.ecsdl.org/content/159/9/H740?related-
urls=yes&legid=jes;159/9/H740 [39] G. J. Janz and N. P. Banal, “Molten salts data: Diffusion coeffi-
cients in single and multi-component salt systems,” J. Phys. Chem. Ref. Data, vol. 11, no. 3, pp. 505–693, 1982. [Online]. Available:
http://scitation.aip.org/content/aip/journal/jpcrd/11/3/10.1063/1.555665 [40] M. Sosna,
G. Denuault,
R. W. Pascal,
R. D. Prien,
and
M. Mowlem, “Development of a reliable microelectrode dissolved oxygen
sensor,” Sens. Actuators B, Chem., vol. 123, no. 1, pp. 344–351,
2007. [Online]. Available: http://www.sciencedirect.com/science/article/
pii/S0925400506006083 [21] M. A. Py and R. R. Haering, “Structural destabilization induced
by
lithium
intercalation
in
MoS2
and
related
compounds,”
Can. J. Phys.,
vol. 61,
no. 1,
pp. 76–84,
1983. [Online]. Available: http://www.nrcresearchpress.com/doi/abs/10.1139/p83-013#. VM9AU9LWKpc [41] D. K. Corrigan,
E. O. Blair,
J. G. Terry, A. R. Mount, and
A. J. Walton, “Enhanced electroanalysis in lithium potassium eutectic
(LKE) using microfabricated square microelectrodes,” Anal. Chem.,
vol. 86, no. 22, pp. 11342–11348, Nov. 2014. [Online]. Available:
http://pubs.acs.org/doi/abs/10.1021/ac5030842 [22] A. Relf, D. Corrigan, C. L. Brady, J. G. Terry, A. J. Walton, and
A. R. Mount, “Robust microelectrodes in molten salt analysis,” ECS
Trans., vol. 50, no. 11, pp. 105–109, 2013. [Online]. Available:
http://ecst.ecsdl.org/content/50/11/105.abstract [23] M. J. Madou, “Lithography,” in Fundamentals of Microfabrication,
2nd ed. Boca Raton, FL, USA: CRC Press, 2002, ch. 1, p. 1. [24] O. Hammerich, “Methods for studies of electrochemical reactions,” in
Organic Electrochemistry, 4th ed. New York, NY, USA: Marcel Dekker,
2001, ch. 2, p. 133. Ewen O. Blair received the M.A. (Hons.) degree
in physics and philosophy from the University of
Aberdeen, Aberdeen, U.K., in 2012. He is cur-
rently pursuing the Ph.D. degree in fabrication and
optimization of durable electrochemical sensors for
molten salts on the EPSRC funded by the REFINE
project. Ewen O. BLAIR et al.: DEVELOPMENT AND OPTIMIZATION OF DURABLE MICROELECTRODES Blair received the M.A. (Hons.) degree
in physics and philosophy from the University of
Aberdeen, Aberdeen, U.K., in 2012. He is cur-
rently pursuing the Ph.D. degree in fabrication and
optimization of durable electrochemical sensors for
molten salts on the EPSRC funded by the REFINE
project. [25] P. Masset, R. J. M. Konings, R. Malmbeck, J. Serp, and J.-P. Glatz,
“Thermochemical properties of lanthanides (Ln = La, Nd) and actinides
(An = U, Np, Pu, Am) in the molten LiCl–KCl eutectic,” J. Nucl. Mater., vol. 344, nos. 1–3, pp. 173–179, 2005. [Online]. Available:
http://www.sciencedirect.com/science/article/pii/S0022311505002217 He was involved in the characterization of novel
photoconductive materials during his time with the
University of Aberdeen. [26] Y.-L. Liu
et
al.,
“Electrochemical
extraction
of
samarium
from
LiCl-KCl
melt
by
forming
Sm-Zn
alloys,”
Electrochim. Acta,
vol. 120,
pp. 369–378,
Feb. 2014. [Online]. Available:
http://www.sciencedirect.com/science/article/pii/S0013468613025218 [27] M. Mohamedi,
N. Kawaguchi,
Y. Sato,
and
T. Yamaura,
“Electrochemical study of the mechanism of formation of the surface
alloy of aluminum–niobium in LiCl–KCl eutectic melt,” J. Alloys
Compounds, vol. 287, nos. 1–2, pp. 91–97, 1999. [Online]. Available:
http://www.sciencedirect.com/science/article/pii/S0925838899000201 Damion
K. Corrigan
is
currently
a
PDRA
with
the
School
of
Chemistry,
University
of
Edinburgh, Edinburgh, U.K. His research experience
lies mainly in the areas of electrochemical and
optical
sensing
technologies. He
has
spent
two years with the Division of Pathway Medicine,
Edinburgh Royal Infirmary, Edinburgh, under the
co-supervision
of
Prof. Mount,
working
on
the development of a point of care compatible
electrochemical sensor for the rapid detection of
MRSA. Damion
K. Corrigan
is
currently
a
PDRA
with
the
School
of
Chemistry,
University
of
Edinburgh, Edinburgh, U.K. His research experience
lies mainly in the areas of electrochemical and
optical
sensing
technologies. He
has
spent
two years with the Division of Pathway Medicine,
Edinburgh Royal Infirmary, Edinburgh, under the
co-supervision
of
Prof. Mount,
working
on
the development of a point of care compatible
electrochemical sensor for the rapid detection of
MRSA. He was with the Electrochemistry Group, University of Southampton,
Southampton, U.K., where he was involved in a project using microstructured
electrodes for the optical detection and electrochemical
discrimination
of
DNA
sequences,
before
moving
to
Edinburgh. He
received
the
Ph.D. degree
in
bioanalytical
chemistry
from
Cranfield
University,
Bedford, U.K., which was funded by GSK, and was involved in the
development
of
sensor
systems
to
address
specific
purity
problems
associated
with
large-scale
pharmaceutical
manufacture. IX. CONCLUSION [14] K. Štulík, C. Amatore, K. Holub, V. Marecek, and W. Kutner,
“Microelectrodes. Definitions, characterization, and applications,” Pure
Appl. Chem., vol. 72, no. 8, pp. 1483–1492, 2000. [Online]. Available:
http://pac.iupac.org/publications/pac/pdf/2000/pdf/7208x1483.pdf This systematic study of layer material combinations for
the manufacture of MS compatible microelectrodes has high-
lighted a number of important issues and challenges. Effective
stress relief is shown to prevent thin film delamination, use
of an electrochemically inert adhesion layer prevents loss of
electrode metal, and the use of stoichiometric silicon nitride
as the top insulator provides effective passivation. The results
of the material testing is summarised in table 1. [15] M. J. Moorcroft, N. S. Lawrence, B. A. Coles, R. G. Compton,
and L. N. Trevani, “High temperature electrochemical studies using a
channel flow cell heated by radio frequency radiation,” J. Electroanal. Chem., vol. 506, no. 1, pp. 28–33, Jun. 2001. [Online]. Available:
http://www.sciencedirect.com/science/article/pii/S0022072801004685 [16] K. T. Chiang and L. Yang, “Development of crevice-free electrodes
for multielectrode array sensors for applications at high tempera-
tures,” Corrosion, vol. 64, no. 10, pp. 805–812, Oct. 2008. [Online]. Available:
http://www.nace.org/cstm/Store/Product.aspx?id=00950ecd-
b924-dc11-94f4-0017a4466950 This paper has described the first microelectrode device
capable of operating over extended periods in the chem-
ically harsh environment of LKE at 500 °C. The impact
of this technology is therefore highly suited to online
monitoring in MS and with the prospect of pyrochemical
processing of nuclear fuel becoming a widely adopted tech-
nique, there is the potential for significant impact. We are
currently developing sensors to enable real time monitoring
for process control in flowing MS media and in stirred reaction
vessels. [17] K. T. Chiang,
L. Yang,
R. Wei,
and
K. Coulter,
“Develop-
ment of diamond-like carbon-coated electrodes for corrosion sen-
sor applications at high temperatures,” Thin Solid Films, vol. 517,
no. 3, pp. 1120–1124, Dec. 2008. [Online]. Available: http://www. sciencedirect.com/science/article/pii/S0040609008009887 [18] K. S. Ujjal,
F. Marken,
B. A. Coles,
R. G. Compton,
and
J. Dupont, “Microwave
activation
in ionic liquids
induces
high
temperature–high speed electrochemical processes,” Chem. Commun.,
vol. 24, pp. 2816–2817, Oct. 2004. [Online]. Available: http://pubs.rsc. org/en/Content/ArticleLanding/2004/CC/B410655E#!divAbstract 1353 JOURNAL OF MICROELECTROMECHANICAL SYSTEMS, VOL. 24, NO. 5, OCTOBER 2015 JOURNAL OF MICROELECTROMECHANICAL SYSTEMS, VOL. 24, NO. 5, OCTOBER 2015 1354 Anthony
J. Walton
(SM’88)
is
currently
a
Professor of Microelectronic Manufacturing with
the School of Engineering, University of Edinburgh,
Edinburgh, U.K. Over the past 25 years, he has been
actively involved with the semiconductor industry in
a number of areas associated with silicon processing
that includes both integrated circuit technology and
microsystems. In particular, he has been intimately
involved in the development of technologies and
their integration with CMOS. He played a key role
in setting up the Scottish Microelectronics Centre,
purpose-built facility for research and development
He has authored over 350 papers. Anthony
J. Walton
(SM’88)
is
currently
a
Professor of Microelectronic Manufacturing with
the School of Engineering, University of Edinburgh,
Edinburgh, U.K. Over the past 25 years, he has been
actively involved with the semiconductor industry in
a number of areas associated with silicon processing
that includes both integrated circuit technology and
microsystems. In particular, he has been intimately
involved in the development of technologies and
their integration with CMOS. He played a key role
in setting up the Scottish Microelectronics Centre,
Edinburgh, which is a purpose-built facility for research and development
and commercialization. He has authored over 350 papers. He has received best paper awards from the IEEE TRANSACTIONS ON
SEMICONDUCTOR MANUFACTURING, the Proceedings of the International
Society of Hybrid Manufacturers, the International Journal of Molecular
Sciences, and the IEEE International Conference on Microelectronic Test
Structures (ICMTS), and received the IET Nanobiotechnology Premium
Award. He is a Fellow of the Royal Society (Edinburgh). He served
as the Chairman for a number of conferences, including the European
Solid-State Devices Research Conference in 1994 and 2008, and ICMTS
in 1989 and 2008. He serves on numerous technical committees. He is
an Associate Editor of the IEEE TRANSACTIONS ON SEMICONDUCTOR
MANUFACTURING. Andrew R. Mount is currently a Professor and the
Head of Physical Chemistry with the University of
Edinburgh, Edinburgh, U.K. He was the Royal Soci-
ety of Edinburgh/SEELLD Support Research Fellow. He has authored over 70 papers and holds 10 patents. He has interests and expertise in electrochemical
production, and the combination of spectroscopic
(in particular, fluorescence) and electrochemical
characterization. Andrew R. Mount is currently a Professor and the
Head of Physical Chemistry with the University of
Edinburgh, Edinburgh, U.K. He was the Royal Soci-
ety of Edinburgh/SEELLD Support Research Fellow. BLAIR et al.: DEVELOPMENT AND OPTIMIZATION OF DURABLE MICROELECTRODES degree in solid-state
electronics
from
the
University
of
Manchester
Institute of Science and Technology, Manchester,
U.K. He joined the Institute for Integrated Micro
and
Nanosystems,
University
of
Edinburgh,
Edinburgh, U.K., in 1999, as a Research Fellow. He is currently a Chancellor’s Fellow and Lecturer
with the University of Edinburgh, where his main
area of interest is in the development of More-than-Moore technologies, and
the integration of novel fabrication processes and materials with foundry
CMOS to create smart microsystems. [33] J. P. Uyemura, “Fabrication and layout of CMOS integrated circuits,” in
CMOS Logic Circuit Design. Norwell, MA, USA: Kluwer, 2001, ch. 2,
sec. 2.4.1, pp. 74–77. [34] O. Zohni, G. Buckner, T. Kim, A. Kingon, J. Maranchi, and R. Siergiej,
“Investigating thin film stresses in stacked silicon dioxide/silicon
nitride structures and quantifying their effects on frequency response,”
J. Micromech. Microeng., vol. 17, no. 5, pp. 1042–1051, 2007. [Online]. Available: http://iopscience.iop.org/0960-1317/17/5/026 [35] K. Hai, T. Sawase, H. Matsumura, M. Atsuta, K. Baba, and R. Hatada,
“Corrosion resistance of a magnetic stainless steel ion-plated with tita-
nium nitride,” J. Oral Rehabil., vol. 27, no. 4, pp. 361–366, Apr. 2000. [Online]. Available: http://www.ncbi.nlm.nih.gov/pubmed/10792599 [36] D. Starosvetsky and I. Gotman, “Corrosion behavior of titanium
nitride coated Ni-Ti shape memory surgical alloy,” Biomaterials,
vol. 22, no. 13, pp. 1853–1859, 2001. [Online]. Available: http://www. ncbi.nlm.nih.gov/pubmed/11396890 His work received a number of awards, including the IEEE International
Conference on Microelectronic Test Structures Best Paper Award in 2004,
the IET Nanobiotechnology Premium Award in 2008, and the International
Journal of Molecular Sciences Best Paper Award in 2013. He has over
70 publications. He is a Treasurer of the Scottish Chapter of the IEEE
Electron Devices Society and Region 8 (U.K., Africa, and Middle East), and
an Editor of the EDS Newsletter. g
p
[37] B. K. Yen et al., “Microstructure and properties of ultrathin amorphous
silicon nitride protective coating,” J. Vac. Sci. Technol. A, Vac., Surf.,
Films, vol. 21, no. 6 pp. 1895–1904, Nov. 2003. [Online]. Available:
http://www.slac.stanford.edu/cgi-wrap/getdoc/slac-pub-10008.pdf JOURNAL OF MICROELECTROMECHANICAL SYSTEMS, VOL. 24, NO. 5, OCTOBER 2015 He has authored over 70 papers and holds 10 patents. He has interests and expertise in electrochemical
production, and the combination of spectroscopic
(in particular, fluorescence) and electrochemical
characterization. He has collaborated with the National Nuclear
Laboratory, Cumbria, U.K., for the last 10 years,
as an Electrochemical Consultant on electroanalysis in room and high
temperature molten salt systems. During this period, he has also been a
Principal Investigator and an active member of the management team in
over £9M of successful multidisciplinary projects to develop optical and
electrochemical sensors and devices, directly supervising six PDRAs and
involving dual and multisite supervision. He is the Chair of the RSC
Electrochemistry Group; a member of the RSC Faraday Standing Committee
on Conferences; the Founding Member of the Centre for Materials Science,
Edinburgh; the Edinburgh Materials Microanalysis Centre, Edinburgh; and
the Centre for Science at Extreme Conditions, Edinburgh; and a reviewer
with the Oak Ridge National Laboratory, Oak Ridge, TN, USA. He was the
Chair of the Faraday Discussion 149 (2010). p p
He has received best paper awards from the IEEE TRANSACTIONS ON
SEMICONDUCTOR MANUFACTURING, the Proceedings of the International
Society of Hybrid Manufacturers, the International Journal of Molecular
Sciences, and the IEEE International Conference on Microelectronic Test
Structures (ICMTS), and received the IET Nanobiotechnology Premium
Award. He is a Fellow of the Royal Society (Edinburgh). He served
as the Chairman for a number of conferences, including the European
Solid-State Devices Research Conference in 1994 and 2008, and ICMTS
in 1989 and 2008. He serves on numerous technical committees. He is
an Associate Editor of the IEEE TRANSACTIONS ON SEMICONDUCTOR
MANUFACTURING.
|
https://openalex.org/W2903196264
|
https://periodicos.ufsc.br/index.php/zeroseis/article/download/1980-4512.2018v20n38p397/37738
|
Portuguese
| null |
Entre a letra da lei e os desafios da prática: tensões e contradições da Educação Infantil carioca
|
Zero-a-seis
| 2,018
|
cc-by
| 8,492
|
1 Doutora em Educação pela Universidade Federal Fluminense. Professor Adjunto I da Universidade Federal do Estado do Rio de
Janeiro. Coordenadora do grupo FRESTAS de pesquisa. E-mail: adrianne.ogeda@gmail.com
2 Graduada em Pedagogia pela Universidade do Estado do Rio de Janeiro. Diretora Adjunta de uma escola pública da Prefeitura
do Rio de Janeiro; Professora de Educação Infantil da Rede Municipal do Rio de Janeiro; Pedagoga e membro do grupo de
pesquisa FRESTAS ligado a UNIRIO. E-mail: michaduda@yahoo.com.br
3 Especialista em Educação Infantil pela UFRJ. Graduada em Pedagogia pela Universidade do Estado do Rio de Janeiro. Professora
de Educação Infantil da rede Municipal do Rio de Janeiro e membro do grupo FRESTAS de pesquisa, vinculado a UNIRIO. E-mail:
laneoliveirasilva@hotmail.com Entre a letra da lei e os desafios da prática:
tensões e contradições da Educação Infantil
carioca Resumo: A formação de professores se insere dentro do tema mais amplo das políticas
de infância no Brasil e evidencia relações tensas entre as diretivas legais para o
campo da Educação Infantil e a realidade de sua assunção no cotidiano das
instituições educativas voltadas para esse segmento. Esse artigo tem como objetivo
evidenciar alguns aspectos referentes às políticas de formação docente, expressos nos
documentos que a normatizam, como por exemplo, os tempos necessários para
planejamento dos professores em exercício e as possibilidades e impossibilidades de
sua realização. Partindo da contextualização da história das políticas voltadas para a
Educação Infantil, com ênfase na formação do professor, buscamos articular os
desafios e tensões presentes na concretização das conquistas legais nas práticas
cotidianas das creches e pré-escolas, tomando por meio da investigação de alguns
elementos da realidade do Município do Rio de Janeiro. Palavras-chave: Educação Infantil. Formação de Professores. Políticas Educacionais. DOI: http://dx.doi.org/10.5007/1980-4512.2018v20n38p397 DOI: http://dx.doi.org/10.5007/1980-4512.2018v20n38p397 Artigos Adrianne Ogêda Guedes1
Michelle Dantas Ferreira2
Edilane Oliveira Da Silva3 Keywords: Child education. Teacher training. Education Policy. 4 A ideologia de gênero diz respeito à ideia de que os dois sexos — masculino e feminino — são construções culturais e soc
portanto, os papéis atribuídos aos gêneros precisam ser problematizados. 6 Kramer (2003) enfatiza a intrínseca relação entre educar e cuidar, argumentando que a Educação Infantil não pode ser
compreendida como uma instância de aprendizagem que só instrui, tampouco como um lugar apenas de guarda e proteção. O
cuidado com o outro deve se fazer presente no ato de educar, independentemente do nível de ensino em que se está atuando. marcas dos processos de socialização das mulheres que, na nossa sociedade, caracterizam-se por identificarem-se
domésticas e a maternagem, características menos valorizadas culturalmente o que, consequentemente se desdo
valorização de quem trabalha com as crianças pequenas (GUIMARÃES, ARENHART E SANTOS, 2017) Between the letter of the law and the challenges
of practice: tensions and contradictions of
Carioca Early Childhood Abstract: The training of teachers is part of the broader theme of childhood policies in
Brazil and highlights tense relations between the legal directives for the field of Early
Childhood Education and the reality of their assumption in the daily life of educational
institutions focused on this segment. This article aims to highlight some aspects related
to teacher education policies, expressed in the documents that regulate it, such as the
time needed for the planning of the teachers in exercise and the possibilities and
impossibilities of their realization. With the emphasis on teacher education, we seek to
articulate the challenges and tensions present in the achievement of legal
achievements in the day-to-day practices of kindergartens and pre-schools, taking the
reality of the State of Rio de Janeiro January as the locus of analysis. 397 ISSNe 1980-4512 | v. 19, n. 36 p.493-504 | jan-jun 2018 Entre a letra da lei e os desafios da prática: tensões e contradições da Educação Infantil carioca Entre a letra da lei e os desafios da prática: tensões e contradições da Educação Infantil carioca Introdução Educação Infantil, história, memória e políticas p
p p
g
p
p
5 Estudos como o de Cerisara (2002) destacam que a identidade profissional da educadora de creche constitui-se no feminino e traz 5 Estudos como o de Cerisara (2002) destacam que a identidade profissional da educadora de creche constitui-se no fem
marcas dos processos de socialização das mulheres que, na nossa sociedade, caracterizam-se por identificarem-se com a
domésticas e a maternagem, características menos valorizadas culturalmente o que, consequentemente se desdobra nu
valorização de quem trabalha com as crianças pequenas (GUIMARÃES ARENHART E SANTOS 2017) (
)
q
p
s processos de socialização das mulheres que, na nossa sociedade, caracterizam-se por identificarem-se com as quest
e a maternagem, características menos valorizadas culturalmente o que, consequentemente se desdobra numa me
o de quem trabalha com as crianças pequenas (GUIMARÃES, ARENHART E SANTOS, 2017) Educação Infantil, história, memória e políticas Se o processo de formação não está aberto à criação, à inovação e à participação dos
atores, fica difícil sustentar um conhecimento com sentido para os professores e para as
crianças. (KRAMER E NUNES, 2007, p. 445) A A história da formação do professor da Educação Infantil no Brasil é razoavelmente recente, pois
até meados do século XX não havia uma preocupação com a educação desta faixa etária, e,
portanto, a necessidade de qualificação destes profissionais não era uma questão relevante. As
primeiras iniciativas voltadas para a educação da criança de 0 a 6 anos no Brasil são marcadas
por uma perspectiva assistencialista, preparatória e instrumental. A Até a década de 1980 a educação das crianças pequenas no Brasil, era de responsabilidade única e
exclusiva das famílias. Esta concepção muda apenas com a constituição de 1988, que em seu Artigo 227
afirma a Educação Infantil como direito da criança. Definindo no artigo 208 como se dará esse
oferecimento: “O dever do Estado com a Educação será efetuado mediante a garantia de (...) atendimento
em creches e pré-escolas às crianças de zero a seis anos de idade”. A partir da Carta Magna de 1988, as responsabilidades com as crianças pequenas passam a ser,
portanto, compartilhadas entre Estado e Família. A partir desta mudança, são pensadas uma série de
políticas públicas para esta faixa etária. No entanto, como dito anteriormente, ainda não havia uma
especificidade no que dizia respeito à formação dos profissionais para este segmento. Os profissionais
ainda eram escolhidos a partir de critérios imprecisos e marcados por uma ideologia de gênero4 que
consideram ser mãe, ser cuidadosa e gostar de criança como atributos suficientes para a docência5. A contar dos anos de 1990 diversos campos da sociedade se mobilizaram em busca de uma
redefinição dos objetivos da Educação Infantil, onde o educar e o cuidar fossem assumidos em sua
indissociabilidade6 e, principalmente, que suas especificidades fossem atendidas, visando o 398 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 Adrianne Ogêda Guedes
Michelle Dantas Ferreira
Edilane Oliveira Da Silva desenvolvimento em todas as áreas do conhecimento. Essa foi uma luta que contou, em especial, com as
“Universidades e institutos de pesquisa, sindicatos de educadores e organizações não governamentais”
(OLIVEIRA, 2002, p.117). Educação Infantil, história, memória e políticas Para atender a essas novas demandas, seria necessário a formação de
profissionais capacitados, pois não havia mais lugar apenas para assistência ou para “cuidadoras” de
crianças, sem um pressuposto educativo. desenvolvimento em todas as áreas do conhecimento. Essa foi uma luta que contou, em especial, com as
“Universidades e institutos de pesquisa, sindicatos de educadores e organizações não governamentais”
(OLIVEIRA, 2002, p.117). Para atender a essas novas demandas, seria necessário a formação de
profissionais capacitados, pois não havia mais lugar apenas para assistência ou para “cuidadoras” de
crianças, sem um pressuposto educativo. A Lei de Diretrizes e Bases 9.394/96 (LDB) é um marco principalmente para as questões ligadas a
Educação Infantil, que passa a estar sob a responsabilidade da Secretaria Municipal de Educação (SME)
no Rio de Janeiro. Além disso, traz reformas, direcionamentos e obrigações em relação a este segmento, e
principalmente, metas para a formação dos profissionais. Com a LDB (BRASIL, 1996), a Educação
Infantil passou a fazer parte do sistema nacional de ensino, como primeira etapa da Educação Básica,
tendo como finalidade o desenvolvimento integral da criança de 0 a 6 anos de idade. Por isso, começa-se a
pensar em uma formação específica para os profissionais que nela atuam ou atuarão, bem como a
incrementar a discussão sobre a especificidade pedagógica da ação docente voltada para esse segmento
(GUIMARÃES, ARENHART E SANTOS, 2017). No artigo 13 da LDB (BRASIL, 1996, p. 6) são determinadas as seguintes funções do professor
da Educação básica: Participar do planejamento das propostas pedagógicas da Instituição; elaborar e
cumprir plano de trabalho de acordo com a proposta pedagógica trabalhada pelo
sistema de ensino; zelar pela aprendizagem; cumprir as horas aulas; buscar estratégias
para recuperação dos alunos com pouco rendimento e colaborar com atividades de
articulação entre a escola e comunidade. Desta forma, o professor tem funções específicas dentro desta concepção de profissional da
Educação Básica. No entanto, o texto não contempla as especificidades do trabalho na Educação Infantil,
e consequentemente, das funções deste profissional, principalmente para instituições que possuem
especificidades próprias – atendimento em horário integral, necessidades de cuidados e educação para
crianças pequenas e bebês. Educação Infantil, história, memória e políticas O artigo 62, da lei supracitada, define que a formação do profissional apto para atuar na Educação
Básica: Básica: [...] far-se-á em nível superior, em curso de licenciatura, de graduação plena, em
universidades e institutos superiores de educação, admitida, como formação mínima
para o exercício do magistério na educação infantil e nos 5 (cinco) primeiros anos do
ensino fundamental, oferecida em nível médio na modalidade Normal. (Redação dada
pela Lei nº 12.796, de 2013). No entanto, antes da LDB essas formações não abordavam temáticas específicas do cotidiano das
instituições educacionais deixando de lado questões como o cuidado e a educação dos bebês ou a
proposta pedagógica de escolas de horário integral (MACHADO, 2000). No artigo 63, a lei enfatiza que
Institutos Superiores de Educação oferecerão a formação em nível superior para os profissionais, da
seguinte forma: 399 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 Entre a letra da lei e os desafios da prática: tensões e contradições da Educação Infantil carioca Entre a letra da lei e os desafios da prática: tensões e contradições da Educação Infantil carioca I - cursos formadores de profissionais para a educação básica, inclusive o curso normal
superior, destinado à formação de docentes para a educação infantil e para as primeiras
séries do ensino fundamental; II - programas de formação pedagógica para portadores de diplomas de educação
superior que queiram se dedicar à educação básica; II - programas de formação pedagógica para portadores de diplomas de educação
superior que queiram se dedicar à educação básica;
III - programas de educação continuada para os profissionais de educação dos diversos
níveis. III - programas de educação continuada para os profissionais de educação dos diversos
níveis. O artigo citado acima explicita a necessidade de uma formação em nível superior. Porém, no que
se refere à Educação Infantil ainda admitir-se-á a formação em Ensino Médio na modalidade Normal,
dando um prazo de dez anos para se adequar à nova legislação. Arce (2001, p.170) nos mostra que a docência na Educação Infantil foi acontecendo de acordo
com a educação que era oferecida, “fortemente impregnada do mito da maternidade, da mulher como
rainha do lar, educadora nata”, ou seja, sem uma preocupação de fato educativa como a entendida nos
dias de hoje, em que se busca efetivar as conquistas legais, os direitos expressos em leis, para que se
materializem na prática. Educação Infantil, história, memória e políticas Para isso se torna imprescindível conhecer as particularidades desse público, de
sua faixa etária, dos profissionais imersos nas instituições e como os direitos garantidos em lei são
vivenciados no cotidiano do município do Rio de Janeiro. As conquistas do campo da Educação Infantil a partir da Constituição Federal e da Lei de
Diretrizes e Bases da Educação Nacional (9.394/1996) aqui enunciadas, se afirmam em um conjunto de
leis que as acompanham e/ou as sucedem, tais como o Estatuto da Criança e do Adolescente (ECA)
(BRASIL, 1990); a Resolução CEB nº 1, de 07 de abril de 1999 (BRASIL, 1999) que homologa as
Diretrizes Nacionais para a Educação Infantil; a Resolução nº 5, de 17 de dezembro de 2009, que fixa as
Diretrizes Curriculares Nacionais para a Educação Infantil (DCNEI) (BRASIL, 2009) e a lei de nº 13.257,
de março de 2016 (Marco Legal) (BRASIL, 2016) que dispõe sobre as políticas públicas para a primeira
infância, para garantir, dentre outros fins, o atendimento profissional devidamente qualificado às
especificidades das crianças de 0 a 6 anos: Art. 10. Os profissionais que atuam nos diferentes ambientes de execução das políticas
e programas destinados à criança na primeira infância terão acesso garantido e
prioritário à qualificação, sob a forma de especialização e atualização, em programas
que contemplem, entre outros temas, a especificidade da primeira infância, a estratégia
da intersetorialidade na promoção do desenvolvimento integral e a prevenção e a
proteção contra toda forma de violência contra a criança. Outros tantos documentos que versam sobre as muitas dimensões que envolvem esse
atendimento, tais como: “Orientações para a organização da sala na Educação Infantil: ambiente para a
criança criar, mexer, interagir e aprender” (SME, 2013) que trata dos espaços físicos; “A avaliação na
Educação Infantil” (SME, 2013) que discute a temática que o nomeia; “Planejamento na Educação
Infantil: cadernos pedagógicos” (SME, 2011) que discorre sobre a temática do planejar, do registro e das
rotinas; “Orientações para profissionais da Educação Infantil” (SME, 2010) que aborda questões relativas
ao cuidado, com o ambiente, com a criança e com a comunidade e “Orientações ao Professor de Pré-
escola I e II” (SME, 2013) voltado para a Pré-escola, que apresenta o trabalho com as apostilas e sugere os
conhecimentos a serem adquiridos na Educação Infantil nessa modalidade. Esses documentos são 400 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 ISSNe 1980-4512 | v. 20, n. 38 p. Educação Infantil, história, memória e políticas 397-411| jul-dez 2018 Adrianne Ogêda Guedes
Michelle Dantas Ferreira
Edilane Oliveira Da Silva Adrianne Ogêda Guedes
Michelle Dantas Ferreira
Edilane Oliveira Da Silva Adrianne Ogêda Guedes
Michelle Dantas Ferreira
Edilane Oliveira Da Silva elaborados pela Secretaria e vêm norteando as práticas na Educação Infantil da Rede Municipal, ao menos
na perspectiva do que é desejável alcançar para a qualidade do trabalho desse segmento. Nesse artigo, cuja ênfase recai sobre as demandas cotidianas do atendimento às crianças da
Educação Infantil e da formação em serviço dos professores na realidade vivenciada no município do Rio
de Janeiro, daremos destaque a algumas políticas que foram garantidas na teoria, mas que ainda possuem
entraves para que se concretizem na prática, como o direito a 1/3 de planejamento garantido pela Lei
11.738 de 2008 em seu Artigo 2º, inciso 4º, ao determinar que “na composição da jornada de trabalho,
observar-se-á o limite máximo de 2/3 (dois terços) da carga horária para o desempenho das atividades de
interação com os educandos”. Isso é reafirmado pelo Parecer CNE/CEB nº 18 de 2012. Com isso, o
professor tem 1/3 de seu planejamento destinado a atividades que não envolvam o contato direto com as
crianças, podendo cumpri-lo em atividades extraclasse, dentro da instituição, sozinho ou com seus pares. Essa nos parece uma questão central, pois diz respeito a garantia dos espaços para reflexão sobre as
próprias práticas, planejamento e organização do trabalho pedagógico, afirmando a necessária
intencionalidade pedagógica no contexto educacional. Outro aspecto que destacamos diz respeito ao excessivo número de crianças por turma neste
segmento, pois apesar da existência da Resolução da SME nº 1427, de 24 de outubro de 2016 que
regulamenta a matrícula, encontramos casos de total discrepância. A diversidade de cargos que diferem em
nomenclaturas, carga horária e salário, mas que possuem as mesmas atribuições, causando uma série de
tensões entre os profissionais que atuam, muitas vezes juntos, em unidades escolares de Educação Infantil
no município do Rio de Janeiro também é um grande nó que necessita ser debatido. O foco que daremos nas linhas que seguem é apresentar o panorama da situação da rede
municipal do Rio de Janeiro no tocante as três grandes questões anunciadas acima, discutindo e tomando
a legislação como referência e alguns dos estudos que abordam a temática, sublinhando a sua importância
para que a Educação Infantil se consolide como alicerce fundamental para as crianças de 0 a 6 anos. ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 Pelas Infâncias Cariocas: políticas e direitos na formação docente Quanto à formação do professor de educação infantil, tanto a inicial quanto a
continuada devem merecer a mesma atenção dispensada à dos demais níveis de ensino. Com isonomia no tratamento administrativo e pedagógico, aliada à urgente valorização
profissional, pode-se evitar que as conquistas, ainda que tímidas, consagradas na atual
Lei de Diretrizes e Bases da Educação nacional, transformem-se em letra morta. (SALLES E RUSSEF, 2003, p. 90) Os cursos de extensão de curta duração, denominados “Infâncias Cariocas nas creches e pré-escolas:
interações e brincadeiras” foram elaborados no intuito de dar continuidade à proposta de formação docente
que já vinha acontecendo, desde 2013, na Faculdade de Educação da Universidade Federal do Estado do
Rio de Janeiro (UNIRIO). O Infâncias Cariocas – assim chamado no decorrer de suas aulas – foi
composto por três cursos simultâneos: Crianças da Natureza, Lugares do Corpo e Políticas do Cotidiano. Com 32
horas de duração cada, divididas em oito encontros semanais, compondo três turmas de 50 alunos em
cada, prevendo um percurso de capacitação e formação docente a 150 profissionais da área da Educação
Infantil e áreas afins, entre maio e julho de 2017. Os três cursos foram, prioritariamente, destinados a
professoras das redes públicas e conveniadas do Estado do Rio de Janeiro, mas atendendo também,
educadores de redes comunitárias, escolas particulares, estudantes e outros profissionais interessados nas
questões de educação voltadas à infância (SCHAEFER, TIRIBA E GUEDES, 2017). As temáticas elegidas interrogavam sobre a macropolítica que mascara a coerção micropolítica
cotidiana, aprisionando os corpos e desconectando-os da natureza, em flagrante desrespeito às leis
brasileiras, que, por sua vez, embasam e empoderam as lutas que visam uma educação de qualidade para as
crianças brasileiras. A metodologia de trabalho dos cursos integrava a garantia permanente de espaços de
interlocução em que a palavra circulasse e houvesse compartilhamento de experiências. Nesse sentido, a
cada encontro, a temática em foco provocava a que todos os participantes trouxessem aspectos das
realidades que vivenciavam, o que nos permitiu compreender mais amplamente as tensões e questões que
atravessam o cotidiano de escolas e professores. Para que pudéssemos compreender como as questões que destacamos nesse artigo se davam na
realidade dos integrantes do “Infâncias Cariocas”, solicitamos ao final do curso, que respondessem a um
questionário cujas questões versavam sobre o 1/3 do planejamento, a lotação de turmas e a multiplicidade
de cargos e salários com nomenclaturas e denominações diferentes. Educação Infantil, história, memória e políticas Para
embasar a pesquisa, validar nosso interesse pelos pontos escolhidos e sustentar as informações que
apresentamos, dialogamos, por meio de questionários aplicados ao final do curso de extensão “Infâncias
Cariocas nas creches e pré-escolas: interações e brincadeiras”, com um grupo de vinte e nove professores da rede
pública de ensino do Rio de Janeiro, que nos permitiram conhecer as especificidades que cada realidade
apresenta e a forma como cada uma dessas questões é vivenciada no cotidiano das instituições. A escolha desses três pontos se deu por sua importância e relevância para o funcionamento das
instituições educacionais da rede municipal do Rio de Janeiro como um todo, especialmente na Educação
Infantil devido a própria história do seu surgimento e da importância que lhe é atribuída, que diz muito
sobre as concepções que o Estado têm acerca da Educação. Além disso, esses foram assuntos que
mobilizaram os professores que integraram os cursos de extensão propostos pelo projeto de extensão
Infâncias Cariocas, oferecido por um grupo de profissionais ligados ao Núcleo Infância Arte e Natureza
(NINA) da Universidade Federal do Estado do Rio de Janeiro (UNIRIO), no primeiro semestre de 2017. Percebendo o quão polêmica e complexa é a realidade da Educação Infantil no município do Rio de 401 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 Entre a letra da lei e os desafios da prática: tensões e contradições da Educação Infantil carioca Entre a letra da lei e os desafios da prática: tensões e contradições da Educação Infantil carioca Janeiro, decidimos nos aprofundar nessas questões, dialogando com a legislação, com autores que
abordam o tema, mas sobretudo com os profissionais que atuam cotidianamente nessa realidade. na prática? O município do Rio de janeiro se constitui como a maior rede de ensino público da América
Latina. Com um total de 1.537 Unidades Escolares divididas em: 1.018 escolas de Ensino Fundamental e
519 Unidades de Educação Infantil, distribuídas em Creches e pré-escolas. São 641.655 alunos
matriculados na Rede, sendo 142.884 crianças na Educação Infantil, subdivididos em: 59.776 em Creches
e 83.108 em Pré-escolas. Com relação ao corpo docente, possui um total geral de 41.302 professores. Destes, 14.337 são Professores I, 16.824 como Professores II, 4.994 Professores de Ensino Fundamental
(PEF) e 5.144 Professores de Educação Infantil (PEI). Há ainda um total de 15.191 funcionários de apoio
administrativo, sendo 5.501 Agentes de Educação Infantil. Devido a seu tamanho, estrutura-se em um nível central que é a Secretaria Municipal de Educação
(SME), subdividida em setores denominados Gerências que tratam de assuntos específicos, tais como:
Educação, Educação Infantil, Recursos Humanos, Infraestrutura, Mídia, entre outras, e onze
Coordenadorias Regionais de Educação (CREs) responsáveis por micro regiões que têm a função de fazer
a intermediação entre as propostas da SME e as escolas, com o objetivo de garantir sua concretização e
execução nas instituições de ensino. Antes de nos atermos as especificidades dos fatos e tensões que todo esse tamanho e diversidade
geram na Educação Infantil, faz-se necessário explicar as diferentes instituições e as nomenclaturas e
funções que cada um desses cargos citados possuem. Como já foi dito anteriormente, a Educação Infantil é a primeira etapa da Educação Básica e é
oferecida em duas modalidades: Creche, para crianças de 0 a 3 anos e 11 meses e Pré-escola, para crianças
de 4 a 5 anos e 11 meses. As Creches possuem suas unidades específicas. A Pré-escola, porém, pode ser
oferecida em Espaços de Desenvolvimento Infantil (EDI) e Centros Integrados de Educação Pública
(CIEPs). O primeiro contempla o atendimento da creche e pré-escola no mesmo espaço físico, sendo
definido pela Prefeitura do Rio de Janeiro, por meio de documento específico, como: [...] um novo modelo público de atendimento à primeira infância e tem como objetivo
principal realizar atendimento educativo às crianças entre 3 meses e 5 anos e 6 meses,
por meio de uma proposta pedagógica que reconheça e valide a integralidade da criança,
isto é, as suas necessidades físicas e de crescimento, psicológicas e emocionais,
educativas e cognitivas, assim como seus desejos e interesses. Pelas Infâncias Cariocas: políticas e direitos na formação docente Foram enviados um total de cento e
cinquenta questionários – entre email e distribuição nas turmas do curso. Porém desse total, tivemos
retorno de trinta e três. Para este trabalho selecionamos exclusivamente as respostas dos professores que
fazem parte do quadro de funcionários do município do Rio do Janeiro e que atuam diretamente na
Educação Infantil. Sendo assim, quatro questionários não foram considerados: dois de professoras da rede
privada e dois de graduandas de Pedagogia que ainda não estão trabalhando em instituições escolares. 402 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 Adrianne Ogêda Guedes
Michelle Dantas Ferreira
Edilane Oliveira Da Silva na prática? (SME, 2010) Já os CIEP’s são instituições que têm sua arquitetura tombada e, portanto, não podem sofrer
alterações estruturais. Antigamente funcionavam em regime integral, ou seja, as crianças entravam às
7h30min. e saíam às 17h, fazendo quatro refeições (café da manhã, almoço, lanche da tarde e jantar) e
tendo acesso a atividades diversificadas como recreação, Educação Física, Teatro, Dança, Artes, entre
outras. Hoje, a grande maioria atua em Turno Único, onde as crianças permanecem por sete horas diárias
e fazem três refeições (café da manhã, almoço e lanche). Normalmente, comporta turmas do Ensino
Fundamental I, porém alguns ainda conservam turmas de Educação Infantil que não conseguiram ser 403 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 Entre a letra da lei e os desafios da prática: tensões e contradições da Educação Infantil carioca absorvidas pelas EDI’s do entorno, seja pela falta de infraestrutura adequada ou pela grande demanda da
comunidade local. No que tange às nomenclaturas e cargos, temos o Professor I, que é aquele habilitado com
Licenciatura Plena para lecionar uma disciplina específica (Artes, História, Geografia, Matemática,
Português, Ciências, Inglês, Espanhol, Educação Musical e Educação Física) e que pode ter sua carga
horária de 16h ou 40h semanais, dependendo do concurso prestado. Esses profissionais atuam em escolas
de Ensino Fundamental II (6º ao 9º ano), excetuando-se os professores de Educação Física, Artes e
Educação Musical que podem atuar da Educação Infantil ao 9º ano, dependendo das demandas da Rede e
de sua lotação. O Professor II é o habilitado com formação mínima exigida de Ensino Médio na modalidade
Normal, para atuar com crianças da Educação Infantil ao 5º ano do Ensino Fundamental I, que ingressou
em concurso anterior ao ano de 2013. Sua carga horária é de 22h30min. semanais, podendo atuar em
escolas de horário parcial, que possui dois turnos: manhã e tarde. Antes do cargo de Professor de Ensino
Fundamental (PEF) este profissional podia também atuar em escolas de horário integral quando possuísse
duas matrículas, fizesse Dupla Regência (DR) ou dividindo a turma com outro professor que também
fosse PII. Hoje, no entanto, com a criação do cargo de PEF que já tem a carga horária de 40h, esses
profissionais foram realocados, em sua maioria, para instituições que funcionam em horário parcial. ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 na prática? Entre eles: não estar a menos de dez/cinco anos de se aposentar – essa
variação se deveu a mudanças no texto no decorrer do processo –; permanecer na mesma Unidade
Escolar por mais dez/cinco anos em função de regência, ou seja, sem poder assumir nenhum cargo
gratificado (Coordenação ou Gestão) ou por indicação (Sala de Leitura e Recurso); ter feito Dupla
Regência em anos anteriores. Além disso, havia prioridade para os profissionais que já atuassem há muitos
anos na mesma instituição e que tivessem maior tempo de serviço, exceto aqueles que estivessem próximo
da aposentadoria. Com isso, “cria-se” mais uma nomenclatura e um novo tipo de profissional, que é o PII
migrado, que passa a ganhar como o PEF – caso tenha nível superior – e ter a mesma carga horária. Porém, difere dele ao poder atuar na Educação Infantil e por ter que cumprir as exigências que o processo
de migração impôs. Algumas dessas especificações podem ser vistas na Lei nº 5623 de 1º de outubro de 2013 que
dispõe sobre o Plano de Cargos, Carreiras e Remuneração dos funcionários da Prefeitura do Rio de
Janeiro, que também apontam as diferenças salariais que esses cargos possuem, mesmo em situações em
que a função desempenhada e a carga horária são as mesmas, como é o caso do Professor de Ensino
Fundamental (PEF) e o Professor de Educação Infantil de 40h. (PEI), diferindo-se apenas a habilitação
mínima exigida no momento do concurso. Ou então, o que consideramos mais grave, a diferença salarial
entre o PEF e o PII com duas matrículas. O primeiro tem uma carga horária semanal de 40h. e ganha mais
do que o segundo que tem que fazer semanalmente, no total, 45h. Essa diferença também é proveniente
da habilitação exigida em concurso, mesmo possuindo as mesmas atribuições no desempenhar das
funções. Ao olharmos somente para a Educação Infantil, foco desse trabalho, percebemos que as
contradições estão presentes mesmo nessa micro esfera. Segundo a Resolução da SME nº 1427, de 24 de
outubro de 2016, as Unidades de Educação Infantil podem ter um funcionamento Integral, em que as
crianças passam nove horas diárias na instituição (7h30min. às 16h30 min.) ou em Turno Único quando a
jornada diária for de sete horas (7h30min. às 14h30min.); parcial, com jornada de 4 horas e 30 minutos
que pode se dar no período da manhã (7h30min. na prática? O Professor de Ensino Fundamental (PEF) é aquele que tem habilitação de nível superior e que
atua do 1º ao 9º ano do Ensino Fundamental com carga horária semanal de 40h. Tanto pode ministrar
uma disciplina específica, como já citado anteriormente, quanto atuar com crianças de seis a dez anos,
ministrando todas as disciplinas que concernem ao Ensino Fundamental I. A orientação inicial era que
esses profissionais fossem lotados em escolas de Turno Único, devido sua carga horária. No entanto, há
PEF’s lotados em escolas de turno parcial que acabam por fazer uma complementação que equivale a
meia Dupla Regência, já que o horário dessas instituições ultrapassam sua carga horária. Existem ainda
escolas de turno parcial em que o PEF se recusa a trabalhar além de sua carga horária e essa diferença de
cerca de duas horas – do horário de saída do professor que é às 15h30min. até o horário de saída da turma
que é às 17h30min. – é suprida por membros da Gestão, Professor de Sala de Leitura e/ou Professor de
Educação Física e Artes. O Professor de Educação Infantil (PEI) é o que fez concurso específico para atuar na Educação
Infantil (Creche e Pré-escola), tendo como exigência mínima ser habilitado em nível médio modalidade
Normal. Sua carga horária pode ser de 22h30min., ou 40h. semanais, dependendo do concurso prestado. Os profissionais que possuem carga horária de 40h e atuam em Creches e alguns EDIs que funcionam em
horário integral de 9 horas, tem que fazer uma complementação, ou seja, precisam cumprir uma hora a
mais todos os dias na Unidade, chegando a 45h por semana. Isso acontece por que a carga horária do
professor é menor que a permanência das crianças nessas unidades. Em 2014 foi aberto, para os professores que possuíam carga horária de 22h30min., o processo de
migração, que os “converteria” para 40h. Havia uma série de quesitos a serem preenchidos para que 404 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 Adrianne Ogêda Guedes
Michelle Dantas Ferreira
Edilane Oliveira Da Silva Adrianne Ogêda Guedes
Michelle Dantas Ferreira
Edilane Oliveira Da Silva realmente isso acontecesse. na prática? às 12h) ou da tarde (13h às 17h30min.) ou em ambos,
quando algumas turmas estão em período integral e outras em horário parcial, dependendo da sua
estrutura e da demanda da Comunidade na qual está inserida. Em sua maioria possuem um Diretor Geral,
um Diretor Adjunto e em alguns casos, um Professor Articulador, que em Creches e EDI’s atua como um
Coordenador Pedagógico. Algumas instituições podem ter outro Diretor Adjunto, como é o caso dos
CIEP’s, mas isso depende do quantitativo total de alunos matriculados. Há ainda nesse espaço os Agentes
de Educação Infantil (BRASIL, 2013) que atuam juntamente com o Professor Regente nas turmas. Esse
profissional que inicialmente era designado como Agente auxiliar de Creche (BRASIL, 2005) tem como
habilitação mínima exigida o Ensino Fundamental completo e sua carga horária é de 40h semanais, sendo
30h em atividades em classe e 10h em atividades extraclasse: 2h em atividades coletivas e 8h de atividades
individuais. . 405 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 Entre a letra da lei e os desafios da prática: tensões e contradições da Educação Infantil carioca Entre a letra da lei e os desafios da prática: tensões e contradições da Educação Infantil carioca No tocante à composição das turmas, a SME dispôs da Resolução nº 1427, de 24 de outubro de
2016, que trata dentre outras coisas, dos critérios para a organização das turmas em todos os segmentos da
Educação Básica. Segundo esse documento, a composição das turmas da Creche dar-se-ão da seguinte
forma: 1.1.1. As turmas do Berçário serão formadas por 25 (vinte e cinco) crianças na faixa
etária de 6 (seis) meses a 1 (um) ano e 11 (onze) meses. 1.1.1. As turmas do Berçário serão formadas por 25 (vinte e cinco) crianças na faixa
etária de 6 (seis) meses a 1 (um) ano e 11 (onze) meses. (
)
(
)
(
)
1.1.3. As turmas do Maternal I serão formadas por 25 (vinte e cinco) crianças na faixa
etária de 2 (dois) anos a 2 (dois) anos e 11 (onze) meses. (
)
(
)
(
)
1.1.4. As turmas do Maternal II serão formadas por 25 (vinte e cinco) crianças na faixa
etária de 3 (três) anos a 3 (três) anos e 11 (onze) meses. na prática? O documento afirma ainda que quando houver uma demanda maior de crianças a serem
matriculadas na instituição, haverá um acréscimo de dez por cento desse total de matrículas em turma,
para suprir as necessidades locais. Com isso, as turmas poderão ter no máximo, segundo a Resolução,
vinte e oito crianças. Poderá também haver a inclusão de até duas crianças com deficiência e transtorno
global do desenvolvimento. Quando isso acontecer, o quantitativo total deverá ser reduzido em duas
crianças para cada criança incluída, mas mantendo-se o número de adultos correspondente a vinte e cinco
crianças. As turmas da Educação Infantil na modalidade Pré-escola seguem o mesmo critério de formação. Designa-se Pré-escola I as turmas formadas por crianças de 4 a 4 anos e 11 meses e Pré-escola II as
compostas por crianças na faixa etária de 5 a 5 anos e 11 meses. Ambas deverão ter um quantitativo de
vinte e cinco crianças, podendo sofrer acréscimo de dez por cento, o que eleva o quantitativo para vinte e
oito crianças. A inclusão de crianças com deficiência e transtorno global do desenvolvimento será feita da
mesma forma que na Creche, podendo ser incluídas até duas crianças por turma e quando feito, o
quantitativo deverá diminuir em dois para cada crianças incluída. Essa, no entanto, é uma realidade que não acontece na maioria das instituições do município do
Rio. Muitas são as Comunidades que contam com poucas instituições de Educação Infantil acabando por
não dar vasão ao número de crianças que necessitam de matrícula, principalmente após a alteração da
LDB (BRASIL, 1996) feita por meio da Lei nº 12.796, de 4 de abril de 2013 que traz obrigatoriedade de
matrícula em Unidades Escolares a partir dos quatro anos de idade. O mesmo quadro se dá na Creche,
visto que a necessidade de ingresso no mercado de trabalho por parte das famílias se faz cada vez mais
crescente, o que leva os responsáveis a procurarem por essas instituições. Em muitos casos, para buscar
atender pelo menos o mínimo de quatro horas exigidas por lei (BRASIL, Seção II, Art. 31, 1996), as
instituições dividem seu horário em dois turnos, dobrando sua capacidade de atendimento. Analisando as respostas dos questionários aplicados no grupo de professores do “Infâncias
Cariocas”, evidenciou-se que todas as turmas encontram-se com quantitativo dentro do estipulado na
Resolução, mesmo aquelas que possuem crianças incluídas. ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 na prática? No entanto, salientamos que essa é uma
amostragem que não reflete necessariamente a condição geral de uma Rede tão grande como a municipal
do Rio de Janeiro. Para apurar mais amplamente essa questão, seria necessário um número maior de
respondentes. 406 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 Adrianne Ogêda Guedes
Michelle Dantas Ferreira
Edilane Oliveira Da Silva O excessivo número de crianças em sala compromete o trabalho do professor que não consegue
muitas vezes ter uma atenção individualizada, uma escuta e olhar mais sensível para o que é particular. O
educar e cuidar também sofre prejuízos, pois um número maior de crianças demanda mais tempo, o que
interfere em toda a rotina da instituição e, consequentemente, no trabalho do professor. Da mesma forma
as propostas compartilhadas com as crianças necessitam abarcar o maior número de crianças visto que
muitas Unidades não possuem uma estrutura física que comporte, por exemplo, a divisão de variados
grupos dentro da mesma sala de aula. Sem contar com a violência do entorno que é um impeditivo para
que professores utilizem áreas externas da escola, quando elas existem. Outra grande polêmica refere-se ao cumprimento do horário de 1/3 para planejamento e outras
atividades sem atuação com crianças. Mas o que a legislação fala a respeito desse 1/3? Porque é
importante que ele seja cumprido? Segundo a Lei 11.738, de 16 de julho de 2008, em seu art. 2º, inciso 4º, “na composição da
jornada de trabalho, observar-se-á o limite máximo de 2/3 (dois terços) da carga horária para o
desempenho das atividades de interação com os educandos”. Isso significa que os outros 1/3 destinam-se
a atividades extra aulas, definidas no Parecer CNE/CEB nº 18/2012 como “horários dedicados à
preparação de aulas, encontros com pais, com colegas, com estudantes, reuniões pedagógicas, didáticas [...]
necessário[s] para a melhoria da qualidade do ensino e também para a redução das desigualdades
regionais”. Na prática isso significa que dentre as 40 horas semanais que o professor deve estar com as
crianças, 13 horas e 30 min. na prática? devem ser utilizadas para planejamento, formação, reuniões e outras
atividades sem que o profissional esteja atuando diretamente com elas, o que possibilita que o professor se
prepare para o trabalho que realizará com as crianças, pense nas questões que são importantes para o seu
grupo, selecione materiais, conheça o acervo da própria instituição, troque com seus pares, tenha um olhar
diferenciado e individualizado para as crianças e busque aprimorar seus conhecimentos em formações no
próprio espaço escolar. Infelizmente, sabemos que esse não é um direito consolidado na prática na maioria das
instituições públicas de ensino do município do Rio de Janeiro, principalmente naquelas que atendem
especificamente ao segmento da Educação Infantil. Ao analisarmos os questionários, pudemos constatar
que apenas oito professores atuam em escolas que têm o 1/3 garantido; seis professores não têm esse
direito garantido de forma alguma; uma alegou não estar apta a responder por atuar como Agente de
Educação Infantil e catorze tem o direito garantido em parte, o que significa dizer que tem um tempo
garantido que não é o total estipulado em Lei, mas também não ficam sem tempo nenhum. Pelas
respostas percebemos ainda que a maioria consegue garantir um tempo quando a turma está em atividades
como Educação Física e Artes, ou sob a regência do Professor Articulador ou Sala de Leitura, o que se
aplica apenas para crianças que estão na pré-escola, uma vez que a maioria das Creches não possui esses
profissionais. Essas, no entanto, são medidas que variam de instituição para instituição e de CRE para CRE,
pois há Coordenadorias que não disponibilizam, por exemplo, professor de Artes para a Educação 407 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 Entre a letra da lei e os desafios da prática: tensões e contradições da Educação Infantil carioca Infantil, o que interfere nessa organização interna para que se tente garantir o horário de 1/3 de
planejamento nessas instituições. Essas variações indicam disputas de poder entre as CREs e as escolas e demonstram a fragilidade
dessa Rede e os contrassensos que a acometem, pois a própria SME, por meio da Circular E/SUBE/CED
n° 11 de 15 fevereiro de 2012 faz a distribuição dessa carga horária e ainda aponta algumas possiblidades
de atividades para serem realizadas com as crianças para que essa proposta se concretize. na prática? O que impede
então que isso se dê concretamente na prática? O documento não só reconhece tal direito como organiza o horário com os diferentes parceiros
com o objetivo de trocar e enriquecer o planejamento pedagógico. Os parceiros citados são os Agentes de
Educação Infantil, que atuam cotidianamente em sala com as crianças e o Professor Articulador, que
possui um olhar ampliado em relação à escola. Além de “garantir” algumas horas em que o professor
reflete sozinho. Portanto, percebemos que há um olhar para a reflexão sobre o trabalho pedagógico na
Educação Infantil, no qual é reconhecida a importância do planejamento com os diferentes parceiros. Mas
é fundamental refletirmos sobre as concepções definidas pelo Município acerca de planejamento. A SME,
por meio da Gerência de Educação Infantil, formulou um documento denominado Planejamento na
Educação Infantil – Cadernos Pedagógicos (SME, 2011), que tem como objetivo guiar as práticas
pertinentes a este segmento e apresentar as concepções que orientam o nível central. Nesse documento o
planejamento é visto como resultado de um processo reflexivo que norteia a prática e que está vinculado
ao registro do trabalho e a observação das crianças de forma individual e coletiva (SME, 2011, p. 11). O planejamento, então, ganha vida se construído coletivamente. Os educadores que convivem
com um grupo de crianças mais de perto têm um olhar voltado para o coletivo e para cada criança em si. Compartilhar essas observações e impressões individuais enriquece o planejamento e o trabalho na
Educação Infantil. Os professores articuladores poderão contribuir com um olhar mais amplo,
relacionando o Projeto Político Pedagógico (PPP) da instituição com o planejamento da turma. O
professor da turma e os Agentes de Educação Infantil (AEI) pensarão em ações para o grupo todo, para
pequenos grupos e para algumas crianças em especial, pois é necessário que esse planejamento atinja a
todos e a cada um, visando proporcionar às nossas crianças um ambiente acolhedor, alegre e sereno, com
condições favoráveis de aprendizagem. (Circular E/SUBE/CED n° 11, 2012) Observamos que para que esse planejamento se concretize é fundamental o encontro, as reflexões
e ações, lembrando que no ano de 2012, quando a circular foi publicada, enfatizava que aos poucos esse
direito seria “gradativamente ampliado de acordo com a disponibilidade de recursos humanos da rede”
(Circular E/SUBE/CED n. 11, 2012, p.1). na prática? Hoje, no ano de 2017, temos uma situação ainda muito
próxima do que tínhamos em 2012, já que tais direitos ainda não são assegurados na prática pela rede. Considerações finais Considerações finais 408 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 Adrianne Ogêda Guedes
Michelle Dantas Ferreira
Edilane Oliveira Da Silva Adrianne Ogêda Guedes
Michelle Dantas Ferreira
Edilane Oliveira Da Silva O presente trabalho refletiu sobre como a letra da lei se materializa ou não, nas práticas de
professores que fazem parte do município do Rio de Janeiro. Ao pensarmos na história da
profissionalização deste docente, pudemos constatar o quão complexa se apresentou, especialmente no
que diz respeito às garantias de alguns direitos adquiridos, mas que de fato não estão concretizados na
rede do Rio de Janeiro. Ao ouvirmos os professores a respeito de 1/3 do planejamento observamos o quanto às leis que
garantem no papel as conquistas descritas no artigo estão bem distantes do que acontece no chão da
escola. Algumas instituições, porém, conseguem assegurar e contemplar aos professores que lá atuam, seu
direito pleno. Diante dessa diversidade presente nas respostas aos questionários, que refletem as diferentes
realidades possíveis dentro de uma mesma rede, pensamos no porquê e no como. Porque algumas
Unidades conseguem, e outras não? Como garantir esse direito a todas? Nesse contexto se mostra fundamental a garantia de espaços para que as atividades extraclasses
aconteçam, favorecendo assim a reflexão individual desse professor e aquela que acontece quando ele está
com seus pares, voltada para as práticas referentes à Educação Infantil, pois acreditamos que desta
maneira possamos construir juntos um entendimento coletivo das nossas especificidades. Além disso, faz-
se urgente que o Munícipio do Rio de Janeiro busque estratégias que possam garantir os direitos dos
profissionais da educação para que assim seja construída uma educação de qualidade, que valorize a
formação e respeite as conquistas legais adquiridas. O respeito à legislação que garante um tempo adequado para que o professor planeje e se forme, a
organização de turmas que tenham o quantitativo determinado pelas Portarias e Resoluções e um plano de
cargos e salários justo e coerente é uma forma de valorização da Educação Infantil e do profissional que
nela atua, assim como dos outros segmentos. É um direito adquirido que contribui para a melhoria das
condições de trabalho e num fazer pedagógico que tem um investimento maior, de tempo, reflexão,
estudo e elaboração. Referências Bibliográficas ARCE, Alessandra. Documentação oficial e o mito da educadora nata na educação infantil. Caderno de
Pesquisa, São Paulo, n. 113, jul. 2001. BIZZO, Kátia; TIRIBA, Léa; GUEDES, Adrianne Ogêda. Infâncias Cariocas: Corpo, Conexão com a
Natureza e Empoderamento Político. Trabalho apresentado no Seminário Fala Outra Escola,
Campinas/SP, 2017. BRASIL. Constituição (1988). Constituição da República Federativa do Brasil. Brasília: Senado, 1988. __________. Lei nº 8.069 de 13 de julho de 1990. Dispõe sobre o Estatuto da Criança e do Adolescente e
dá outras providências. Diário Oficial da União, Brasília, 16 jul. 1990. __________. Lei nº 8.069 de 13 de julho de 1990. Dispõe sobre o Estatuto da Criança e do Adolescente e
dá outras providências. Diário Oficial da União, Brasília, 16 jul. 1990. 409 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 Entre a letra da lei e os desafios da prática: tensões e contradições da Educação Infantil carioca __________. Lei nº 9.394 de 20 de dezembro de 1996. Estabelece as diretrizes e bases da educação
nacional. Diário Oficial da União, Brasília, 23 dez. 1996. ___________. Lei nº 3985 de 8 de abril de 2005. Cria no Quadro Permanente do Poder Executivo do
Município do Rio de Janeiro a categoria funcional que menciona e dá outras providências. Diário Oficial
do Município, Rio de Janeiro, 12 abr. 2005. ___________. Lei nº 5623 de 1 de outubro de 2013. Dispõe sobre o Plano de Cargos, Carreiras e
Remuneração dos funcionários da Secretaria Municipal de Educação e dá outras providências. Diário
Oficial do Município, Rio de Janeiro, 2 out. 2013. ___________. Lei nº 11.738 de 16 de julho de 2008. Regulamente a alínea “e” do Inciso III do caput do
art. 60 do Ato das Disposições Constitucionais Transitórias, para instituir o piso salarial profissional
nacional para os profissionais do magistério público da educação básica. Diário Oficial da União,
Brasília, 17 jul. 2008. ___________. Lei nº 12.796 de 4 de abril de 2013. Altera a Lei nº 9.394, de 20 de dezembro de 1996, que
estabelece as diretrizes e bases da educação nacional, para dispor sobre a formação dos profissionais da
educação e dar outras providências. Diário Oficial da União, Brasília, 5 abr. 2013. ___________. Lei nº 13.257 de 8 de março de 2016. BRASIL. Constituição (1988). Constituição da República Federativa do Brasil. Brasília: Senado, 1988. Dispõe sobre as políticas públicas para a primeira
infância e altera a Lei nº 8.069, de 13 de julho de 1990 (Estatuto da Criança e do Adolescente), o Decreto-
Lei nº 3.689, de 3 de outubro de 1941 (Código de Processo Penal), a Consolidação das Leis do Trabalho
(CLT), aprovada pelo Decreto-Lei nº 5.452, de 1º de maio de 1943, a Lei nº 11.770, de 9 de setembro de
2008, e a Lei nº 12.662, de 5 de junho de 2012. Diário Oficial da União, Brasília, 9 mar. 2016. ___________. Ministério da Educação. Secretaria de Educação Básica. Diretrizes Curriculares
Nacionais para a Educação Infantil. Brasília: MEC/SEB, 2010. ___________. Ministério da Educação. Conselho Nacional de Educação. Resolução CEB nº 1, de 7 de
abril de 1999. Institui as Diretrizes Curriculares Nacionais para a Educação Infantil. Diário Oficial da
União, Brasília, 13 abr. 1999. ___________. Ministério da Educação. Conselho Nacional de Educação. Parecer CEB nº 18, de 2 de
outubro de 2012. Trata da implementação da Lei nº 11.738 de 2008, que institui o piso salarial profissional
nacional para os profissionais do magistério público da Educação Básica. Diário Oficial da União,
Brasília, 1 ago. 2013. ___________. Ministério da Educação. Conselho Nacional de Educação. Resolução nº 5, de 17 de
dezembro de 2009. Fixa as Diretrizes Curriculares Nacionais para a Educação Infantil: Brasília:
MEC/CNE, 2009. 410 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 Adrianne Ogêda Guedes
Michelle Dantas Ferreira
Edilane Oliveira Da Silva GUIMARÃES, Daniela Oliveira; ARENHART, Deise; SANTOS, Núbia Oliveira. Educação Infantil Pós
LDB/96: identidade docente e formação de professores. Revista Contemporânea de Educação,
UFRJ/Rio de Janeiro, vol. 12, n. 24, maio/ago. 2017. KRAMER, Sonia; NUNES, Maria Fernanda. Gestão pública, formação e identidade de profissionais de
Educação Infantil. Cadernos de Pesquisa, São Paulo, vol. 37, n. 131, p. 423-454, maio/ago. 2007. MACHADO, Maria Lúcia de Almeida. Desafios iminentes para projetos de formação de
profissionais para educação infantil. In: Cadernos de Pesquisa nº 110. São Paulo: Cortez, 2000. OLIVEIRA, Zilma Moraes Ramos. Educação Infantil: fundamentos e métodos. São Paulo: Cortez,
2010. (Coleção docência em formação). SALLES, Fernando Casadei; RUSSEFF, Ivan. Formação continuada do professor de educação infantil e
identidade profissional. In: RUSSEFF, Ivan; BITTAR, Mariluce (Orgs.). Educação Infantil: política,
formação e prática docente. Brasília-DF: Editora Plano/UCDB, vol. I, 2004. SECRETARIA MUNICIPAL DE EDUCAÇÃO. A avaliação na Educação Infantil. Rio de Janeiro,
jul. 2013. ____________. BRASIL. Constituição (1988). Constituição da República Federativa do Brasil. Brasília: Senado, 1988. Espaço de Desenvolvimento Infantil (EDI): modelo conceitual e estrutura. Rio de
Janeiro, fev. 2010. ____________. Orientações para a organização da sala na Educação Infantil: ambiente para a
criança criar, mexer, interagir e aprender. Rio de Janeiro, mar. 2013. ____________. Orientações para a organização da sala na Educação Infantil: ambiente para a
criança criar, mexer, interagir e aprender. Rio de Janeiro, mar. 2013. ____________. Orientações para profissionais da Educação Infantil. Rio de Janeiro, jul. 2010. ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018 ________. Orientações para profissionais da Educação Infantil. Rio de Janeiro, jul. 2010. ____________. Orientações ao Professor de Pré-escola I e II. Rio de Janeiro, 1º semestre de 2013. ____________. Planejamento na Educação Infantil. Cadernos Pedagógicos, vol. I, Rio de Janeiro, fev. 2011. ____________. Planejamento na Educação Infantil. Cadernos Pedagógicos, vol. I, Rio de Janeiro, fev. 2011. ____________. Resolução nº 1427, de 24 de outubro de 2016. Dispõe sobre a estrutura de atendimento,
organização de turmas, horário de funcionamento e Matriz Curricular das Unidades Escolares da Rede
Pública de Ensino da Cidade do Rio de Janeiro, e dá outras providências. Diário Oficial do Município,
Rio de Janeiro, 26 out. 2016. ___________. Circular E/SUBE/CED nº 11, de 15 de fevereiro de 2012. Dispõe sobre horário
Extraclasse Semanal para os profissionais da Educação Infantil – Informações complementares AAC e
PEI. Recebido em: 15/08/2017
Aprovado em: 15/12/2017 411 ISSNe 1980-4512 | v. 20, n. 38 p. 397-411| jul-dez 2018
|
https://openalex.org/W2894093652
|
https://kar.kent.ac.uk/70401/1/nsy085.pdf
|
English
| null |
The social buffering of pain by affective touch: a laser-evoked potential study in romantic couples
|
Social cognitive and affective neuroscience
| 2,018
|
cc-by
| 9,959
|
Licence for this version
CC BY (Attribution) Additional information Versions of research works Versions of Record
If this version is the version of record, it is the same as the published version available on the publisher's web site. Cite as the published version. Author Accepted Manuscripts If this document is identified as the Author Accepted Manuscript it is the version after peer review but before type
setting, copy editing or publisher branding. Cite as Surname, Initial. (Year) 'Title of article'. To be published in Title
of Journal , Volume and issue numbers [peer-reviewed accepted version]. Available at: DOI or URL (Accessed: date). Downloaded from The version of record is available from
https://doi.org/10.1093/scan/nsy085 Kent Academic Repository von Mohr, Mariana, Krahé, Charlotte, Beck, Brianna and Fotopoulou, Aikaterini
(2018) The social buffering of pain by affective touch: a laser-evoked potential
study in romantic couples. Social Cognitive and Affective Neuroscience,
13 (11). pp. 1121-1130. ISSN 1749-5016. Downloaded from Downloaded from
https://kar.kent.ac.uk/70401/ The University of Kent's Academic Repository KAR Enquiries If you have questions about this document contact ResearchSupport@kent.ac.uk. Please include the URL of the record
in KAR. If you believe that your, or a third party's rights have been compromised through this document please see
our Take Down policy (available from https://www.kent.ac.uk/guides/kar-the-kent-academic-repository#policies). Social Cognitive and Affective Neuroscience, 2018, 1121–1130 doi: 10.1093/scan/nsy085
Advance Access Publication Date: 24 September 2018
Original article Mariana von Mohr,1 Charlotte Krahé,2 Brianna Beck,3
and Aikaterini Fotopoulou1 1Research Department of Clinical, Educational and Health Psychology, University College London, London
WC1E 6BT, UK 2Department of Psychology, Institute of Psychiatry, Psychology, and Neuroscience, King’s College
London, London SE5 8AF, UK, and 3Institute of Cognitive Neuroscience, University College London, London
WC1N 3AZ, UK Correspondence should be addressed to Mariana von Mohr, Research Department of Clinical, Educational and Health Psycho
University College London, 1-19 Torrington Place, London WC1E 7HB, UK. E-mail: mariana.ballina.13@ucl.ac.uk /scan/article-abstract/13/11/1121/5106208 by guest on 03 December 2018 Abstract Pain is modulated by social context. Recent neuroimaging studies have shown that romantic partners can provide a potent
form of social support during pain. However, such studies have only focused on passive support, finding a relatively
late-onset modulation of pain-related neural processing. In this study, we examined for the first time dynamic touch by
one’s romantic partner as an active form of social support. Specifically, 32 couples provided social, active, affective (vs active
but neutral) touch according to the properties of a specific C-tactile afferent pathway to their romantic partners, who then
received laser-induced pain. We measured subjective pain ratings and early N1 and later N2-P2 laser-evoked potentials
(LEPs) to noxious stimulation, as well as individual differences in adult attachment style. We found that affective touch from
one’s partner reduces subjective pain ratings and similarly attenuates LEPs both at earlier (N1) and later (N2-P2) stages of
cortical processing. Adult attachment style did not affect LEPs, but attachment anxiety had a moderating role on pain
ratings. This is the first study to show early neural modulation of pain by active, partner touch, and we discuss these
findings in relation to the affective and social modulation of sensory salience. Key words: affective touch; attachment style; laser-evoked potentials; pain; romantic couples on 03 December 2018 Received: 3 March 2018; Revised: 12 September 2018; Accepted: 21 September 2018
© The Author(s) 2018. Published by Oxford University Press.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/),
which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. Introduction mammals, including humans, have adapted to the presence
and active care of other conspecifics, so that our ability to form
and regulate emotions (Atzil and Barrett, 2017) and our sense
of selfhood (Fotopoulou and Tsakiris, 2017) are constituted
on the basis of early social interactions. Thus, according to
such theories, social proximity and active social support may
constitute the default assumption of the human brain (i.e. social
baseline theory; Beckes and Coan, 2011; Coan, 2011) or inherited
‘priors’ in predictive coding accounts (Decety and Fotopoulou,
2015). An ensuing prediction of such theories is that individuals
employ fewer higher-order, self-regulatory psychological and
neural processes when faced with threats in socially supportive Social bonding and support is important for human well-
being (Berscheid, 2003; Ditzen and Heinrichs, 2014; Uchino,
2006). Close social bonds, or attachment relationships, have
long been suggested to serve safety and distress-alleviating
functions (Bowlby, 1969; Mikulincer et al., 2003). Interestingly,
evidence from non-human mammals further suggests that it
is not the mere presence of conspecifics but rather certain
active behaviours (e.g. tactile contact, grooming, licking by
conspecifics) that are important for affective regulation (Nelson
and Panksepp, 1998). Accordingly, recent proposals suggest that © The Author(s) 2018. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/),
which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. 1121 1121 1122
Social Cognitive and Affective Neuroscience, 2018, Vol. 13, No. 11 contexts than when alone (Coan, 2011; Eisenberger et al.,
2007). Indeed, functional neuroimaging studies have shown an
attenuation of neural responses typically implicated in affective
regulation (e.g. dorsolateral prefrontal cortex, anterior insula),
when social support (e.g. hand-holding by a romantic partner
vs a stranger) is provided in the face of physical threat
(Coan et al., 2006), including pain (Eisenberger et al., 2011;
Krahé et al., 2015). given that slow, CT-optimal touch can specifically and without
prior knowledge signal positive emotions and social support
(Kirsch et al., 2017; von Mohr et al., 2017), this manipulation can
be done off-line, i.e. not simultaneously with, but before the
noxious stimulation, to implicitly signal a socially supportive
context to the individual about to receive pain. Introduction Indeed, a recent laser-evoked potential (LEP) study found that
individual differences in adult attachment style (i.e, individual
differences in the perception of social relationships themselves)
determine how a stranger’s slow CT-optimal affective touch
(vs faster and rated as emotionally neutral but otherwise iden-
tical touch), applied before noxious stimulation, affects early
responses to noxious stimuli, namely the N1 component (Krahé
et al., 2016). While there was no main effect of affective touch on
pain in this study and late, cortical responses to pain were not
reliably affected (i.e. there was no modulation of the P2 and the
N2 was only modulated by an interaction between attachment
anxiety and avoidance dimensions), it remains possible that
slow, affective touch provided by a romantic partner, where
social trust and attachment is already established, might also
impact higher-order pain regulation, as captured by later LEP
components, i.e. the N2-P2 complex. A romantic partner’s slow
affective touch can be more powerful as affective touch is central
to intimate, romantic relationships (Suvilehto et al., 2015), and
the regulatory role of touch seems to be mediated by psycholog-
ical intimacy (Debrot et al., 2013). However, the explanatory potential of these neuroimaging
studies is restricted in two important ways that we aim to
address in this study. The first restriction is that such studies
have mostly focused on passive support of one’s partner vs
control conditions of absence of such support, e.g. presence vs
absence (Krahé et al., 2015), static hand-holding vs no or stranger
hand-holding (Coan et al., 2006; Goldstein et al., 2018), viewing
pictures of one’s partner vs a stranger (Eisenberger et al., 2011). In contrast, there are no neuroscientific studies on active forms
of social support from one’s partner, even though in behavioural
studies passive and active support have been found to have
opposite psychological effects on pain (Krahé and Fotopoulou,
2018; Krahé et al., 2013). Moreover, comparisons between sup-
portive vs non-supportive actions (i.e.active support) of the same
support provider have greater experimental control and hence
explanatory power than many of the manipulations of the above
studies, given that they are not subject to confounds such as
social distraction, comfort and familiarity. Accordingly, in this
study we aimed to examine for the first time the effects of
different forms of tactile active social support by one’s romantic
partner on pain. Introduction Downloaded from https://academic.oup.com/scan/article-abstract/13/11/1121/5106208 by guest on 03 December 2018 In sum, the present study goes beyond previous research
to investigate the effects of a form of active social support,
namely affective touch, on subjective and neural responses to
pain in the context of a romantic relationship. Healthy women
received slow, CT-optimal touch by their partners vs faster, CT
non-optimal touch, followed by laser-evoked noxious stimula-
tion, without any other communication between partners. We
measured self-reported pain as well as deflections in the ongoing
electroencephalogram (EEG) time-locked to transient noxious
radiant heat stimulation, namely the N1 and N2-P2 components
(Plaghki and Mouraux, 2005), that can tease apart different stages
of pain processing: the N1 consists of an early deflection peaking
around 160 ms post-stimulus onset and is thought to reflect early
sensory (nociceptive) processing preceding conscious awareness
(Lee et al., 2009; Valentini et al., 2012), whereas the N2-P2 com-
prises a later biphasic complex peaking around 200–350 ms
post-stimulus onset and is considered to reflect the salience
associated with a conscious experience of pain (Lee et al., 2009;
Mouraux and Iannetti, 2009). Using these methods, we sought
to test two main hypotheses. First, we hypothesised that slow
vs fast touch would attenuate subjective pain ratings and LEPs
reflecting both early and later stages of pain processing, namely
the N1 and N2-P2 local peak amplitudes. Second, we expected
such effects to be moderated by individual differences in adult
attachment style as in previous studies on social support and
pain (Hurter et al., 2014; Krahé et al., 2015, 2016; Sambo et al.,
2010), in that affective touch should have the largest effect in
individuals with higher attachment anxiety (who fear of rejec-
tion and seek clear signals of support) and the smallest effect
in individuals with higher attachment avoidance (who prefer to
cope with threat alone). Although there are many ways to provide active support,
recent experimental studies (Kirsch et al., 2017; von Mohr et al.,
2017) and corresponding theoretical reviews (Fotopoulou and
Tsakiris, 2017; Morrison, 2016) suggest that a particular type
of dynamic touch may be a particularly effective and salient
embodied form of communicating active, social support. Introduction Specif-
ically, slow (at 1–10 cm/s velocities), light-pressure (≈0.4 N),
dynamic (moving along the skin) touch has been shown to com-
municate social support (Kirsch et al., 2017) and reduce social
pain (von Mohr et al., 2017) in comparison to faster, but otherwise
identical, active touch. Importantly, it seems that there may be a
dedicated neurophysiological system, the C-tactile (CT) system,
coding this particular type of affective touch (Croy et al., 2016;
Löken et al., 2009; Olausson et al., 2002; Olausson et al., 2008;
Triscoli et al., 2013). Importantly, this type of touch allows us to address the
second major restriction of the existing neuroimaging studies
on partner support during pain. Namely, simultaneous manipu-
lations of social touch (e.g. hand-holding) and pain as in previous
studies (e.g. Goldstein et al., 2018; see also Coan et al., 2006),
does not allow precise inferences about the mechanisms of pain
modulation, given the following: (i) the existing, consciously
known meaning of hand-holding (i.e. it is not clear whether
the observed pain modulation is the outcome of the feeling of
being touched or the knowledge about the meaning of hand-
holding), (ii) the well-known analgesic effects of touch on pain
(Liljencrantz et al., 2017; Mancini et al., 2015) and (iii) the fact that
these studies cannot control for skin-to-skin touch parameters
(e.g. pressure of handholding, movement, sweating, tempera-
ture) or distraction effects. In contrast, one could control for
most of these confounds and their interactions by comparing
slow touch, that is known to be mediated by the CT-system
and is typically perceived as pleasant, with faster but otherwise
identical touch, that is known to not activate the CT system
optimally and is typically judged to feel ‘neutral’. Specifically, Materials and measures Tactile stimulation. Two skin areas (9 cm long × 4 cm wide) were
marked on the participant’s right forearm (i.e. stimulation sites). The partner administered the touch to the participant using
a cosmetic make-up brush (Natural Hair blush brush, No. 7,
Boots, UK). The partner was trained to administer each touch
condition by watching a 4 min video and then practicing the
touch on the second experimenter outside the testing room. In
each touch condition, the stroking was administered in four 45
s mini-blocks in an elbow-to-wrist direction (Essick et al., 2010;
Krahé et al., 2016) at slow (3 cm/s, 1 stroke) or fast (18 cm/s,
6 strokes) velocities. The velocities of the slow and fast touch
were chosen as they have been shown to be optimal and non-
optimal, respectively, for targeting CT afferents (Löken et al.,
2009; Gentsch et al., 2015). Further, these same velocities have
also been validated in our previous studies and have revealed
statistically significant differences in their effects on social and
physical pain (Krahé et al., 2016; von Mohr et al., 2017) and the
communication of social support (Kirsch et al., 2017). The 3 s
stroking was alternated with 3 s pauses. Stimulation sites were
also alternated between trials to avoid CT habituation. Adult attachment style. We employed the ECR-R (Fraley et al.,
2000) to measure adult attachment style. This questionnaire is
designed to measure individual differences with respect to the
extent to which individuals are insecure about the responsive-
ness and availability of their romantic partners (i.e, attachment
anxiety) and the extent to which individuals are uncomfortable
with being close and depending on their romantic partners
(i.e. attachment avoidance). The ECR-R consists of 36 items on a
7-point scale and yields continuous scores on attachment anxi-
ety and attachment avoidance dimensions, with higher scores
denoting greater attachment anxiety and avoidance, respec-
tively. The ECR-R is a well-validated measure (Ravitz et al., 2010). In the present sample, Cronbach’s alpha was α = 0.86 for attach-
ment anxiety and α = 0.89 for attachment avoidance. Nociceptive stimulation and subjective pain report. We used an
infrared CO2 laser stimulation device with a wavelength of
10.6 μm (SIFEC, Ferrières, Belgium) to deliver noxious radiant
heat stimulation. The laser stimulus (80 ms duration, spot
diameter of 6 mm) was applied to the dorsum of participant’s
left hand, changing the stimulation site between consecu-
tive applications. Design Our
within-subjects
design
comprised
two
experimental
conditions: slow touch (3 cm/s; affective CT-optimal touch) and
fast touch (18 cm/s; neutral, non-CT optimal) administered by
the partner—with the order of these conditions counterbalanced
across participants. Outcome measures were subjective pain
ratings and N1, N2 and P2 local peak amplitudes. The moderating
effect
of
adult
attachment
style
was
examined
using
a
questionnaire that measures the degree of attachment anxiety
and avoidance that individuals may experience in close, adult
romantic relationships [Experiences in Close Relationships-
Revised (ECR-R); Fraley et al., 2000]. Downloaded from https://academic.oup.com/scan/article-abstract/13/11/1121/5106208 by guest on 03 December 2018 Downloaded from https://academic.oup.com/scan/article-abstract/13/11/1121/5106208 EEG
data
were
processed
and
prepared
for
statistical
analysis using EEGLAB/ERPLAB toolboxes for MATLAB (R2015b)
(see
Supplementary
Material
for
EEG
data
pre-processing
details). Average waveforms per condition were computed
(experimental
trials
only). For
each
waveform,
the
peak
amplitude of the N1, N2 and P2 were measured as follows: the
N1 was measured at the central electrode contralateral to the
stimulated side (C6), referenced to Fz (Krahé et al., 2015, 2016). It
was defined as the most negative deflection following stimulus
onset and preceding the N2 wave (Lee et al., 2009). The N2 and
P2 were measured at the vertex (Cz) referenced to the average
of P9 and P10 (electrodes close to the mastoids; Luck, 2014). The N2 and P2 were defined as the most negative and positive
deflection, respectively, after stimulus onset (Lee et al., 2009). In accordance with prior literature reporting that the earliest
neural activity associated with laser stimulation occurs after
120 ms (Valentini et al., 2012), no deflection occurring before
120 ms after stimulus onset was selected as the peak (Krahé
et al., 2015). Data exclusion due to technical issues resulted in a
final sample size of 29 (N = 29) for N1 and 29 (N = 29) for N2 and
P2 analyses (see Supplementary Material). Participants Thirty-two couples in a romantic relationship were recruited. We experimentally induced pain in the women (henceforth
‘participants’), while their partners delivered the (slow affective, M. von Mohr et al. 1123 M. von Mohr et al. 1123 reported pain intensity was recorded using a numeric keyboard
on an 11-point scale ranging from 0 (no pinprick sensation) to 10
(extremely painful pinprick sensation). Mean pain ratings for the
experimental stimuli in each block (across the four mini-blocks
for the touch conditions) were used as the measure of subjective
pain report. Data exclusion due to technical issues resulted in a
final sample size of 31 (N = 31). fast neutral) touch. Participants were included if they were right-
handed and had been in their current relationship for over
a year. The mean age of participants and their partners was
M = 24.53 (s.d. = 3.78) and M = 26.31 (s.d. = 4.65), respectively. See Supplementary Material for other inclusion criteria and
sample characteristics. The University College London Research
Ethics Committee approved this study, and the experiment was
conducted in accordance with the Declaration of Helsinki. EEG recording and LEP analyses. The study was carried out using
a BioSemi ActiveTwo EEG system (http://www.biosemi.com;
Biosemi, Amsterdam, The Netherlands) with a 64-electrode cap. A BioSemi analog input box connected to the analog output
of the laser stimulation device was used to record the online
measurement of skin temperature at target laser stimulation
site in register with the EEG recording across the experimental
task. The electrooculography was monitored with a total of
four electrodes located at the outer canthi of both eyes as well
as above and below the right eye. The sampling rate during
recording was 1024 Hz. Materials and measures Using an ascending-descending-ascending
staircase, we identified each participant’s Aδ threshold for
‘pinprick pain’ (i.e. the lowest skin temperature that elicited
a report of ‘pinprick sensation’, which is linked to Aδ fibres; Lee
et al., 2009). The pain threshold (M = 47.65◦C, s.d. = 2.35) was
used to set a mild-to-moderate (but always tolerable) sharp
pinprick sensation (3◦C above threshold, experimental trials)
and no pinprick sensation (2◦C below threshold, distractor trials)
(See Supplementary Material for details). Manipulation checks. As a manipulation check, participants were
asked to rate retrospectively how comfortable they had felt with
the touch velocities delivered by their partner in each condition. These reports were given on a scale ranging from −3 (not at
all comfortable) to 3 (extremely comfortable). We also collected
pleasantness ratings of the touch to make sure participants
perceived slow touch as more pleasant than fast touch. Here,
participants received 12 randomized trials of 3 s stroking at slow Each block consisted of 60 laser stimuli (40 experimental
stimuli and 20 distractor stimuli), presented in pseudorandom
order with an interstimulus interval of 10–15 s. Participant’s self- 1124
Social Cognitive and Affective Neuroscience, 2018, Vol. 13, No. 11 Fig. 1. Experimental design. Our experimental design for the main task included a baseline (no touch) nociceptive block followed by a fast touch or a slow touch
block. The order of the touch (fast or slow) blocks was counterbalanced across participants. The laser stimuli (experimental and distractor trials) were presented in
pseudorandom order with an interstimulus interval of 10–15 s. Downloaded from https://academic.oup.com/scan/article-abstract/13/11/1121/5106208 by guest on 03 December 2018 Fig. 1. Experimental design. Our experimental design for the main task included a baseline (no touch) nociceptive block followed by a fast touch or a slow touch
block. The order of the touch (fast or slow) blocks was counterbalanced across participants. The laser stimuli (experimental and distractor trials) were presented in
pseudorandom order with an interstimulus interval of 10–15 s. (3 cm/s) and fast (18 cm/s) velocities, the same touch velocities as
in the main task. Using a scale ranging from 0 (not at all pleasant)
to 100 (extremely pleasant), participants were asked to rate the
pleasantness of the touch after each trial. Slow and fast touch
ratings were averaged separately for each participant. (∼120 mins), participants were asked retrospectively to rate how
comfortable they felt with the touch provided by their partner in
each condition. Procedure Upon obtaining written informed consent, each participant’s
pinprick pain threshold was determined, and the experimental
and distractor laser intensities to be used in the main task were
set based on this threshold. The experiment consisted of three
laser blocks. Participants did not see or speak with their partners
during any of the blocks, and they were prevented from seeing
the stimulated skin areas through the use of a black box placed
around the stimulated arms. Materials and measures To avoid biasing the results of the main pain task, partici-
pants returned for a second visit, between 3 and 5 days after
the first visit, in which they completed the adult attachment
style questionnaire (ECR-R) and provided pleasantness ratings
for slow and fast touch. Participants were paid £50 for their
(and their partner’s) time and were fully debriefed at the end of
the second visit. Descriptive statistics and manipulation checks Mean adult attachment scores were M = 2.50 (s.d. = 0.75) for
attachment anxiety and M = 2.55 (s.d. = 0.69) for attachment
avoidance (see Supplementary Material, Table S2 for compar-
isons with the general population). Attachment anxiety and
avoidance dimensions were correlated at r = 0.35, P < 0.05. On average, participants reported good relationship quality/
adjustment (M = 25.84, s.d. = 3.18) as measured by the seven-
item Dyadic Adjustment Scale (Sharpley and Rogers, 1984). Relationship quality/adjustment did not correlate with attach-
ment anxiety or avoidance dimensions (see Supplementary
Material). Table 1 presents descriptive statistics for pain ratings
and associated neural responses (N1, N2 and P2 local peak
amplitudes). Main effects. Full model results are presented in Table 2. Sup-
porting our first hypothesis, a significant main effect of touch
condition was found on the subjective pain ratings, N1, N2 and P2
local peak amplitudes. All our effects were in the same direction:
Regarding the pain ratings, participants reported less pain in the
slow touch (M = 3.01, SE
= 0.13) compared to the fast touch
(M = 3.58, SE = 0.13) condition (see Table 2). With respect to
the neural responses associated with pain, the N1 local peak
amplitude was significantly smaller in the slow touch (M = −3
.48 μV, SE = 0.37) compared to the fast touch (M = −4.28 μV,
SE = 0.39) condition; the N2 local peak amplitude was signif-
icantly smaller in the slow touch (M = −5.92 μV, SE = 0.59)
compared to the fast touch (M = −7.65 μV, SE = 0.59) condition,
and the P2 local peak amplitude was significantly smaller in
the slow touch (M = 9.65 μV, SE = 0.80) compared to the fast
touch (M = 12.11 μV, SE = 0.80) condition. No other main effects
were significant (see Table 2). Together, these results suggest that
pain report and associated neural responses were attenuated in
response to slow, affective touch relative to fast, neutral touch
(see Figure 2 for N1, N2-P2 waveforms). Downloaded from https://academic.oup.com/scan/article-abstract/13/11/1121/5106208 by guest on 03 December 2018 As expected, participants reported feeling more comfortable
with slow touch (M = 2.38, s.d. = 1.16), as compared to fast touch
(M = 1.41, s.d. = 1.74) from their partner, t(31) = 3.67, P = 0.001. Participants also reported higher pleasantness in response to
slow touch (M = 72.15, s.d. Plan of statistical analyses All statistical analyses were conducted in STATA (Version 14). As repeated measures (Level 1) were nested within individu-
als (Level 2), multilevel modelling was implemented. For each
outcome variable (pain ratings and N1, N2 and P2 local peak
amplitude), we specified multilevel models with touch condition
(slow touch/fast touch) as a dummy-coded categorical predictor,
attachment avoidance and attachment anxiety as continuous
predictors, and included all interaction terms. We controlled
for pain baseline differences by including them as covariates
(see also Supplementary Material, Table S1 for analyses con-
trolling for relationship quality in our models). All continuous
predictors were mean-centred in order to avoid multicollinearity
issues (Tabachnick and Fidell, 2007). Given that multilevel mod-
els address the multiple comparisons problem and yield more
efficient estimates (Gelman et al., 2012), we did not correct for
multiple comparisons any further in our model. Significant inter-
actions were followed up by examining differences between con-
ditions at low (−1 s.d.), moderate (mean) and high (+1 SD) con-
tinuous attachment style scores (see Aiken and West, 1991 for
testing and interpreting interactions on continuous variables). In the first block, we recorded participant’s EEG while admin-
istering a baseline nociceptive stimulation block (no touch). In the two other blocks, participants received one of the two
stroking velocity conditions (slow or fast touch) from their part-
ner, followed by noxious stimuli (with the order of the stroking
velocity conditions counterbalanced across participants). Each
of these touch blocks was divided into four mini-blocks, alter-
nating tactile stimulation with noxious stimulation (tactile and
noxious stimulation were administered in spatial and temporal
incongruence and asynchrony in order to avoid concurrent mul-
tisensory effects; see Figure 1 for a schematic of the experimen-
tal design). Between each block, there was a 7 min break with
Sudoku and/or crossword puzzle to minimise carryover effects;
in the meantime, the partner was trained with the other touch
velocity she/he was about to deliver to the participant. EEG was
recorded throughout the periods of laser stimulation following
slow and fast touch conditions. At the end of the study visit Table 1. Descriptive statistics and manipulation checks = 13.48), as compared to fast touch
(M = 54.72, s.d. = 14.16), t(31) = −5.89, P < 0.001. Thus, our manip-
ulations were successful in terms of perceived pleasantness and
comfort of the touch. ademic.oup.com/scan/article-abstract/13/11/1121/5106208 by guest on 03 December 2018 Table 2. Slow vs fast touch: multilevel modelling results for all outcome measures
Effect
Dependent
variable
b
SE
P-value
Confidence
intervals
Lower
Upper
Slow touch vs fast touch
N1
−0.97
0.46
0.036
−1.88
−0.06
N2
−2.06
0.76
0.007
−3.54
−0.57
P2
2.85
0.89
0.001
1.12
4.59
Pain ratings
0.62
0.13
<0.001
−0.37
0.86
Attachment anxiety
N1
−0.21
0.54
0.691
−1.28
0.85
N2
−1.35
0.86
0.116
−3.03
0.33
P2
1.61
1.18
0.17
−0.69
3.92
Pain ratings
0.01
0.20
0.97
−0.39
0.41
Attachment avoidance
N1
0.05
0.60
0.939
−1.13
1.22
N2
−0.67
0.96
0.488
−2.56
1.22
P2
−1.45
1.32
0.270
−4.03
1.13
Pain ratings
−0.06
0.22
0.787
−0.49
0.37
Attachment anxiety × attachment avoidance
N1
−0.77
0.81
0.343
−2.36
0.82
N2
−2.70
1.22
0.028
−5.11
−0.29
P2
0.97
1.68
0.564
−2.33
4.27
Pain ratings
−0.02
0.29
0.945
−0.59
0.55
Touch condition × attachment anxiety
N1
1.17
0.64
0.068
−0.09
2.43
N2
1.26
1.04
0.229
−0.79
3.31
P2
−2.01
1.23
0.101
−4.41
0.393
Pain ratings
−0.41
0.18
0.023
−0.76
−0.05
Touch condition × attachment avoidance
N1
0.28
0.74
0.709
−1.17
1.72
N2
2.19
1.17
0.061
−0.10
4.50
P2
1.78
1.37
0.195
−0.91
4.48
Pain ratings
−0.15
0.19
0.442
−0.22
0.51
Touch condition × attachment avoidance × attachment anxiety
N1
0.19
1.05
0.854
−1.87
2.26
N2
1.31
1.50
0.384
−1.64
4.25
P2
−2.22
1.76
0.207
−5.67
1.23
Pain ratings
−0.26
0.26
0.305
−0.76
0.24
Note. Significant main effects and interactions are highlighted in bold. Same pattern of results were observed when controlling for relationship quality
(see Supplementary Material, Table S1). While the interaction between attachment anxiety and attachment avoidance was statistically significant, follow-up
tests were non-significant/trend level (see Supplementary Material, Figure S1). Baseline pain as a covariate was statistically significant across all pain outcomes,
P < 0.05. Table 2. Slow vs fast touch: multilevel modelling results for all outcome measures Note. Significant main effects and interactions are highlighted in bold. Same pattern of results were observed when controlling for relationship quality
(see Supplementary Material, Table S1). While the interaction between attachment anxiety and attachment avoidance was statistically significant, follow-up
tests were non-significant/trend level (see Supplementary Material, Figure S1). Plan of statistical analyses Mean (s.d.) for pain-related outcome measures
Baseline (no touch)
Slow touch
Fast touch
N1 local peak amplitude (μV)
−5.33 (3.27)
−3.50 (2.23)
−4.20 (2.60)
N2 local peak amplitude (μV)
−10.90 (7.09)
−5.92 (4.61)
−7.52 (5.08)
P2 local peak amplitude (μV)
16.02 (10.14)
9.65 (7.48)
11.81 (6.42)
Pain ratings
4.06 (1.73)
3.01 (1.84)
3.58 (1.82) M. von Mohr et al. 1125 M. von Mohr et al. 1125 Results
Descriptive statistics and manipulation checks Note. Significant main effects and interactions are highlighted in bold. Same pattern of results were observed when controlling for relationship quality
(see Supplementary Material, Table S1). While the interaction between attachment anxiety and attachment avoidance was statistically significant, follow-up
tests were non-significant/trend level (see Supplementary Material, Figure S1). Baseline pain as a covariate was statistically significant across all pain outcomes,
P < 0.05. Descriptive statistics and manipulation checks The N1 has been linked
to activation in the operculoinsular and primary somatosensory
cortex (Garcia-Larrea et al., 2003; Valentini et al., 2012), and such
initial cortical coding of noxious afferent inputs is considered an
essential, yet distinct stage of signal processing, different from
later stages that are associated with the conscious aspects of
pain and its affective regulation. Downloaded from https://academic.oup.com/scan/article-abstract/13/11/1121/5106208 by guest on 03 December 2018 Fig. 2. (A) Effect of touch condition on the N2-P2 waveform measured at the
vertex (Cz). (B) Effect of touch condition on the N1 waveform measured at the
contralateral side of stimulation (C6). N1, N2 and P2 local peak amplitude was
significantly smaller in the slow touch compared to the fast touch condition, as
denoted by asterisks. Baseline pain (no touch) as a covariate was statistically
significant across the N1, N2 and P2 local peak amplitude. Touch condition in interaction with adult attachment style. Partially
supporting our second hypothesis, we found a significant touch
condition by attachment anxiety interaction on pain ratings,
b = −0.41, SE = 0.18, P = 0.023, but not on the neurophysiological
outcome measures. Follow-up tests on the pain ratings showed
that the difference between slow and fast touch conditions
was significant for low (b = −0.93, SE = 0.20, P < 0.001) and
moderate attachment anxiety (b = −0.62, SE = 0.13, P < 0.001),
but not for high attachment anxiety (b = −0.31, SE = 0.17,
P = 0.074); see Figure 3. Thus, the higher the attachment anxiety,
the smaller was the difference between slow and fast touch
on pain ratings, i.e. at high levels of attachment anxiety, slow
and fast touch did not differ in terms of their effects on pain
ratings. There was no significant two-way interaction between
attachment dimensions and no three-way interaction of touch
condition, attachment anxiety and attachment avoidance on
the pain ratings, indicating that these results were driven by
the attachment anxiety dimension. Contrary to our second
hypothesis,
the
interaction
between
touch
condition
and
attachment avoidance was non-significant for all outcome
measures. It is unlikely that the LEP downregulation we observed is
based on potential interactions between nociceptive and CT
pathways at the spinal cord level (Liljencrantz et al., 2017;
Mancini et al., 2015), as the tactile and noxious stimulation in
our study were delivered at different times and in different
body locations. Descriptive statistics and manipulation checks Baseline pain as a covariate was statistically significant across all pain outcomes,
P < 0.05. 1126
Social Cognitive and Affective Neuroscience, 2018, Vol. 13, No. 11 Fig. 2. (A) Effect of touch condition on the N2-P2 waveform measured at the
vertex (Cz). (B) Effect of touch condition on the N1 waveform measured at the
contralateral side of stimulation (C6). N1, N2 and P2 local peak amplitude was
significantly smaller in the slow touch compared to the fast touch condition, as
denoted by asterisks. Baseline pain (no touch) as a covariate was statistically
significant across the N1, N2 and P2 local peak amplitude. stages of cortical processing. Contrary to our second hypothesis,
adult attachment style did not affect LEPs as in other social
contexts (Krahe et al., 2016), but one facet of adult attachment
style, namely attachment anxiety, had a moderating role on self-
reported pain. These findings are discussed in more detail below. Regarding our first hypothesis about the role of active, affec-
tive touch on pain reduction, we found such effects on pain
report and LEPs reflecting both early and later stages of cortical
nociceptive processing, namely the N1 and N2-P2 local peak
amplitude. Our findings on the N2-P2 complex, which has been
linked to activity in areas such as the anterior insula and anterior
cingulate cortex (Garcia-Larrea et al., 2003) and to late, conscious
aspects of noxious processing (Lee et al., 2009; Mouraux and
Iannetti, 2009), are consistent with previous neuroimaging stud-
ies on passive social support (e.g. Eisenberger et al., 2011; Krahé
et al., 2015), which found similar downregulation of brain areas
supporting conscious aspects of noxious processing, such as the
anterior insula and dorsal anterior cingulate cortex (Eisenberger
et al., 2011). However, contrary to these studies, our N1 findings
indicate that the effects of active, affective touch may begin at
earlier stages of cortical nociceptive processing. Even though the
N1 wave represents an early stage of sensory processing more
directly related to ascending nociceptive input (Lee et al., 2009;
Valentini et al., 2012), such cortical encoding is already ‘late’ in
the grand scheme of noxious encoding. Descriptive statistics and manipulation checks Instead, given that LEPs have been recently
proposed to detect environmental threat to the body in response
to sensory salient events (Legrain et al., 2011; Mouraux and
Iannetti, 2009), we speculate that affective touch by one’s
romantic partner when applied before noxious stimulation may
reduce the sensory salience of impending noxious stimulation. Salience has various definitions. Here, we use the term to
describe the importance of a stimulus (its weighting in relation
to other factors) for indicating potential or actual threat to the Fig. 3. Touch condition by attachment anxiety effects for pain ratings. Statisti-
cally significant differences are marked by asterisk, P < 0.05. Participant’s self-
reported pain intensity was recorded on an 11-point scale ranging from 0 (no
pinprick sensation) to 10 (extremely painful pinprick sensation). Discussion While passive social support from one’s romantic partner can
have pain-attenuating effects and corresponding modulation of
neural responses (Eisenberger et al., 2011; Goldstein et al., 2018;
Krahé et al., 2015), little is known about the effects of active
partner support on pain. Here, we investigated the effects of
partner CT-optimal touch on pain, given the experimentally
established role of this type of touch in the communication
of positive emotions and social support (Kirsch et al., 2017;
von Mohr et al., 2017). We found that slow, affective vs fast, neu-
tral touch from one’s partner reduces subjective pain ratings and
similarly attenuates LEPs both at earlier (N1) and later (N2-P2) Fig. 3. Touch condition by attachment anxiety effects for pain ratings. Statisti-
cally significant differences are marked by asterisk, P < 0.05. Participant’s self-
reported pain intensity was recorded on an 11-point scale ranging from 0 (no
pinprick sensation) to 10 (extremely painful pinprick sensation). M. von Mohr et al. 1127 body (Legrain et al., 2011) and for inducing related responses
(Garcia-Larrea and Peyron, 2013). Conceptualising the LEP-
related brain activity as being part of a ‘salience network’
(Legrain et al., 2011), we have recently proposed that activity in
this salience network may also be modulated by information
regarding contextual factors from the (social) environment
(Krahé and Fotopoulou, 2018; Krahé et al., 2013; von Mohr and
Fotopoulou, 2018; see also Atlas and Wager, 2012; Büchel et al.,
2014 for the role of expectations and active inference). Although
the precise neurophysiological mechanisms of the effects of
active, affective touch on pain will need to be studied in future
studies, we discuss below four possible explanations of how
the sensory salience of noxious stimulation may have been
moderated in this study. Third, given that CT firing correlates with perceived pleasant-
ness in response to dynamic stroking (Löken et al., 2009), with our
own findings also suggesting increased perceived pleasantness
in response to slow (vs fast) touch, it is possible that affective
touch may reduce the sensory salience of impending noxious
stimulation in a similar way as positive mood-related manip-
ulations (e.g. positive/pleasant pictures, music and odours, see
Villemure and Bushnell, 2002, for a review). Given the importance
for quick and unbiased experimental succession between touch
and pain, we did not collect mood ratings in this experiment. Discussion Instead, we merely examined, as an off-line manipulation check,
the perceived sensory pleasantness of slow and fast touch. Inter-
estingly, the degree to which participants perceived slow and
fast touch to be pleasant was not related to pain modulation in
the corresponding conditions (similar to Krahé et al., 2016; see
Supplementary Material, Table S3). Thus, future studies should
include specific online measures of mood to further explore this
hypothesis. body (Legrain et al., 2011) and for inducing related responses
(Garcia-Larrea and Peyron, 2013). Conceptualising the LEP-
related brain activity as being part of a ‘salience network’
(Legrain et al., 2011), we have recently proposed that activity in
this salience network may also be modulated by information
regarding contextual factors from the (social) environment
(Krahé and Fotopoulou, 2018; Krahé et al., 2013; von Mohr and
Fotopoulou, 2018; see also Atlas and Wager, 2012; Büchel et al.,
2014 for the role of expectations and active inference). Although
the precise neurophysiological mechanisms of the effects of
active, affective touch on pain will need to be studied in future
studies, we discuss below four possible explanations of how
the sensory salience of noxious stimulation may have been
moderated in this study. Downloaded from https://academic.oup.com/scan/article-abstract/13/11/1121/5106208 by guest on 03 December 2018 First, it is well known that pain can be modulated by distrac-
tion. While our study controls various facets of social distraction
better than previous studies (see Introduction), similarly to
other pain modulation studies, we cannot exclude with absolute
certainty that our two touch conditions did not have some differ-
ence in their general attentional demands. For example, neutral
touch delivered at fast speeds might demand greater attention
than slow, affective touch (see also Davidovic et al., 2017 for
an attenuation of the Default Mode Network in response to non-
affective touch). However, we think this is unlikely because if
fast touch was, in fact, more attention grabbing and thus
distracting, we would expect fast vs slow touch to attenuate
pain. Instead, our findings show the opposite pattern and as
discussed below, it is possible that the mechanism by which
affective touch can selectively modulate pain may relate to its
salience. Fourth, CT-optimal touch is a particularly effective form of
communicating embodied (non-verbal) social support (Kirsch
et al., 2017; von Mohr et al., 2017). Discussion Specifically, recent evidence
on this very modality suggests that this particular kind of slow
dynamic touch, but not the faster stroking touch also tested here
as a control condition, conveys positive social intentions such
as social support even in the absence of any other sensory or
social cue (Kirsch et al., 2017). Thus, it is possible that this type
of touch attenuates the saliency of impending noxious stimuli
by signalling the presence of an active, socially supportive envi-
ronment. This interpretation is consistent with recent theories
on the importance of social interactions for the experience and
regulation of emotions, and particularly homeostatic emotions
such as pain (Atzil and Barrett, 2017; Fotopoulou and Tsakiris,
2017). According to such theories, the perception of the social
environment of pain can affect inferential processes about the
perception of these modalities by influencing the weighting of
prior expectations about certain sensory signals vs the signals
themselves in given contexts (Decety and Fotopoulou, 2015;
Krahé et al., 2013; von Mohr and Fotopoulou, 2018; similar to how
non-social expectations influence pain, e.g. Atlas and Wager,
2012; Geuter et al., 2017). Accordingly, affective touch prior to a
noxious stimulus may modulate pain by changing beliefs about
how threatening a noxious stimulus is in a supportive social
context. Future studies should thus also take direct measure
of perceived social support and examine whether the latter
possibility or more general positive mood effects best explain the
effects of CT-optimal touch on pain. In addition, future studies
could elucidate whether the effects of slow, affective touch on
pain are specific to the CT system. For example, could slow touch
to glabrous skin, that does not possess CT fibres (McGlone et al.,
2014), lead to similar effects in romantic couples? Second, given the many factors that can influence pain
when comparing across different socially supporting contexts,
e.g. individual factors, habituation, distraction, mood, social
presence, familiarity and many more, our aim was to compare
directly two specific and well-controlled types of touch, namely
slow and fast stroking on the forearm by the same, familiar
person at different times and in different body locations than
the noxious stimulation. To address this aim optimally, we have
elected to use a within-subjects design, measuring individual
pain measures before any manipulation, and subsequently
counterbalancing order between our two critical conditions. Discussion 11 strangers (Krahé et al., 2016), may be explained by the fact that
individual differences in attachment may have less of a role to
play when there is an existing degree of attachment security
between partners. The latter can be assumed in the partners
of the current study who were in a relationship for at least
12 months, had good relationship quality (see Supplementary
Material) and showed relatively secure attachment in relation
to existing norms on the same measure and our previous study
(see Supplementary Material, Table S2). activation of this system (Löken et al., 2009), the functional role of
this system and its particular, neurophysiological contribution
to our effects remain to be specified by future studies. Third,
we only tested pain in women, while their partners provided
support by touch, to control for gender effects associated with
the perception of touch (Gazzola et al., 2012; Suvilehto et al.,
2015); however, future research is needed to examine whether
the present results extend to men. Finally, the sources of
the N1 and its functional implications remain debated: most
notably, in relation to the precise contribution of the primary
somatosensory cortex and operculoinsular cortex (Iannetti
et al., 2005; Tarkka and Treede, 1993; Valentini et al., 2012) as
well as its implications for perceptual vs pre-perceptual pain
processing at such early stages. To the best of our knowledge, our study was the first to
investigate the effects of active, affective touch on pain in the
context of a romantic relationship. A recent LEP study by our lab
suggests that there are no main effects of affective touch on pain
when a stranger (confederate) administers the touch, but rather
these effects depend on attachment style, mostly modulating
early stages of cortical processing, namely the N1 component. Specifically, even though the N2 mirrored the effects on the
N1, such effects by attachment style (e.g. high anxiety) were
observed only in relation to the other attachment dimension
(e.g. low avoidance), and other later cortical responses to pain
such as the P2 were not affected (Krahé et al., 2016). Thus, here
we extend these findings to suggest that this type of active
embodied social support can modulate not only early (N1) but
also later (N2-P2) stages of cortical processing. Discussion Critically, and
unlike the first study with confederates, these effects on early
and later stages of cortical processing by affective touch were not
moderated by adult attachment style, which may well pertain
to the social context studied in the present study (i.e. romantic
couples). Indeed, a romantic partner’s affective touch can be
more powerful as affective touch is central to intimate, romantic
relationships (Suvilehto et al., 2015; Croy et al., 2016) and the
regulatory role of touch seems to be mediated by psychological
intimacy (Debrot et al., 2013). Downloaded from https://academic.oup.com/scan/article-abstract/13/11/1121/5106208 by guest on 03 December 2018 In sum, we found that active touch administered by the
romantic partner at the optimal velocities of the CT system prior
to noxious laser stimulation at a different body part attenu-
ated subjective pain ratings and neurophysiological responses
to pain, namely the N1, N2 and P2 local peak amplitudes more
than touch administered at non-optimal CT velocities. Such
effects were moderated only by one facet of adult attachment
style (attachment anxiety) and only for subjective ratings of
pain rather than the neurophysiological measures. Our effects
indicate that the analgesic effects of active affective touch may
begin at earlier stages of cortical nociceptive processing, as
reflected by the N1 local peak amplitude, and expand to later,
conscious aspects of noxious processing, as reflected by the N2-
P2 complex and self-reported pain ratings. Given that LEPs have
been recently proposed to detect environmental threat to the
body in response to sensory salient events (Legrain et al., 2011),
we propose that affective touch by one’s romantic partner (when
applied before noxious stimulation) may reduce the sensory
salience of impending noxious stimulation, due to either its
perceived affective or pro-social effects. .oup.com/scan/article-abstract/13/11/1121/5106208 by guest on 03 December 2018 More generally, while there are many ways to provide active
support during pain (e.g. supportive text messages, verbal reas-
surance, social distraction), the current findings are important
given that the only variable manipulated was the velocity of
the touch from the romantic partner. Thus, we demonstrate
that a simple, yet specific embodied interaction can have pain-
attenuating effects without the need for any explicit labelling
by words or pictures. Discussion While this design is optimal for assessing whether pain
responses differ between the two critical conditions, it does
not allow us to disentangle the potential general effects of touch
on pain (beyond the critical manipulation of stroking speed) and
the general effects of condition order on pain. In other terms,
baseline always precedes the two touch conditions, and hence
any general touch effects on pain could be due either to the
touch or the fact that the touch conditions come always after
the baseline condition and hence may be subject to habituation
effects. However, we can say that there is an effect of slow,
affective touch vs fast, neutral touch on pain over and above
any order effects, as their order was counterbalanced across
conditions. It is still possible, however, that fast, neutral touch
may be increasing pain in comparison to slow touch rather
than the other way around. As we previously said, we cannot
disentangle the role of habituation from the role of general touch
in our studies, and hence it is possible that fast, neutral touch
was associated with less habituation than slow touch in our
study. However, we also note that neutral touch has long being
known to reduce rather than increase pain in previous studies
(e.g. see Mancini et al., 2015 for recent study) and future studies
could thus include further speeds to account for the direction of
the observed effects. Turning now to our second study hypothesis about the
potential moderating role of adult attachment style based on
the observation of such effects in different social contexts
(e.g. Krahé et al., 2016), we found such an effect only on subjective
pain ratings and only in relation to adult attachment anxiety. Specifically, the higher the attachment anxiety, the smaller
the effects of slow vs fast touch on self-reported pain. Given
that anxious attachment is associated with craving closeness
and reassurance from others (Hazan and Shaver, 1987), we
hypothesise that any kind of physical contact, in this case slow
or faster touch from one’s partner, is enough to ease attachment
anxiety, signal closeness and hence attenuate self-reported pain. The fact that we did not observe any other attachment effects on
our pain measures as in earlier work on affective touch between 1128
Social Cognitive and Affective Neuroscience, 2018, Vol. 13, No. Funding This work was supported by the European Research Council
Starting Investigator Award (ERC-2012-STG GA313755 to A.F.) and
the National Council on Science and Technology scholarship
(CONACyT CVU-538843 to M.V.M). Author contributions M. von Mohr, L. C. Krahé and A. Fotopoulou developed the
hypothesis and research plan. B. Beck provided guidance on
the laser stimulation device and EEG. M. von Mohr and C. Krahé
analysed the data. M. von Mohr collected the data and wrote
the manuscript, under the guidance of C. Krahé, B. Beck and
A. Fotopoulou. All authors approved the final version of the
manuscript for submission. Discussion Moreover, in comparison to other types
of embodied social support, such as for example hand-holding,
the tactile interaction studied here was manipulated with a
degree of experimental control, at a time different than the
noxious stimulation and tested against control conditions that
involve the same support provider. Therefore, the problematic
comparison between partners and strangers or friends could be
avoided and many confounding factors, such as social proximity,
familiarity and social desirability can be excluded as potential
explanations of our effect. Supplementary data Supplementary data are available at SCAN online. Conflict of interest. None declared. However, despite these methodological advantages, our
study had several limitations. First, the experimental control
of the study limits its ecological validity as typically couples will
use a much richer embodied and verbal interaction to provide
social support. Relatedly, in order to be able to assess the effects
of touch on pain, including LEPs, as well as to avoid concurrent
multisensory effects, the touch was delivered in advance,
and repeated in mini-blocks, of ‘impending’ noxious stimuli. However, it is likely that romantic couples will also use this type
of embodied social support as a soothing, consoling touch during
or after pain and future studies could explore any differences
based on such timescales. Second, while this study examined
the pain-attenuating effects of touch delivered at velocities that
activate the CT system optimally vs velocities of minimal known References Functional dissoci-
ation of stimulus intensity encoding and predictive coding of
pain in the insula. Elife, 6, e24770. Coan, J.A. (2011). The social regulation of emotion. In: Decety, J.,
Cacioppo, J.T., editors, The Oxford Handbook of Social Neuroscience
(pp. 614–23). New York: Oxford University Press. Goldstein, P., Weissman-Fogel, I., Dumas, G., Shamay-Tsoory, S.G. (2018). Brain-to-brain coupling during handholding is associ-
ated with pain reduction. Proceedings of the National Academy of
Sciences. http://doi.org/10.1073/pnas.1703643115. Croy, I., Luong, A., Triscoli, C., Hofmann, E., Olausson, H.,
Sailer, U. (2016). Interpersonal stroking touch is targeted to
C tactile afferent activation. Behavioural Brain Research, 297,
37–40. http://doi.org/10.1016/j.bbr.2015.09.038. Hazan, C., Shaver, P. (1987). Romantic love conceptualized
as an attachment process. Journal of Personality and Social
Psychology,
52(3),
511–24. http://doi.org/10.1037//0022-3514. 52.3.511. Davidovic, M., Göran, S., Håkan, O. (2017). Processing of affective
and emotionally neutral tactile stimuli in the insular cor-
tex. Developmental Cognitive Neuroscience. http://doi.org/10.1016/
j.dcn.2017.12.006. Hurter, S., Paloyelis, Y., Amanda, A.C., Fotopoulou, A. (2014). Partners’ empathy increases pain ratings: effects of per-
ceived empathy and attachment style on pain report and
display. Journal of Pain, 15(9), 934–944. http://doi.org/10.1016/
j.jpain.2014.06.004. Debrot, A., Schoebi, D., Perrez, M., Horn, A.B. (2013). Touch as
an interpersonal emotion regulation process in couples’ daily
lives. Personality and Social Psychology Bulletin, 39(10), 1373–85. http://doi.org/10.1177/0146167213497592. Iannetti, G.D., Zambreanu, L., Cruccu, G., Tracey, I. (2005). Operculoinsular cortex encodes pain intensity at the earliest
stages of cortical processing as indicated by amplitude of laser-
evoked potentials in humans. Neuroscience, 131(1), 199–208. http://doi.org/10.1016/j.neuroscience.2004.10.035. Decety, J., Fotopoulou, A. (2015). Why empathy has a benefi-
cial impact on others in medicine: unifying theories. Fron-
tiers in Behavioural Neuroscience, 8, 457. http://doi.org/10.3389/
fnbeh.2014.00457. Ditzen,
B.,
Heinrichs,
M. (2014). Psychobiology
of
social
support:
the
social
dimension
of
stress
buffering. Restorative
Neurology
and
Neuroscience,
32(1),
149–62. http://doi.org/10.3233/RNN-139008. Kirsch, L.P., Krahé, C., Blom, N., et al. (2017). Reading the mind
in the touch: neurophysiological specificity in the communi-
cation of emotions by touch. Neuropsychologia. http://doi.org/
10.1016/j.neuropsychologia.2017.05.024. Eisenberger, N.I., Master, S.L., Inagaki, T.K., Taylor, S.E., Shirinyan,
D., Lieberman, M.D., Naliboff, B.D. (2011). Attachment figures
activate a safety signal-related neural region and reduce pain
experience. Proceedings of the National Academy of Sciences,
108(28), 11721–6. http://doi.org/10.1073/pnas.1108239108. Krahé, C., Drabek, M.M., Paloyelis, Y., Fotopoulou, A. (2016). Affective touch and attachment style modulate pain: a
laser-evoked potentials study. Philosophical Transactions of
the Royal Society B: Biological Sciences, 371(1708), 20160009. http://doi.org/10.1098/rstb.2016.0009. Krahé, C., Fotopoulou, A. (2018). References pleasant touch. Neuroscience and Biobehavioral Reviews, 34(2),
192–203 http://doi.org/10.1016/j.neubiorev.2009.02.003. Aiken, L.S., West, S.G. (1991). Multiple Regression: Testing and Inter-
preting Interactions, Newbury Park: Sage. Fotopoulou, A., Tsakiris, M. (2017). Mentalizing homeostasis:
the social origins of interoceptive inference-replies to com-
mentaries. Neuropsychoanalysis,
19(1),
71–6. http://doi.org/
10.1080/15294145.2017.1307667. Atlas, L.Y., Wager, T.D. (2012). How expectations shape pain. Neuroscience Letters, 520(2), 140–8. Atzil, S., Barrett, L.F. (2017). Social regulation of allostasis:
commentary on “Mentalizing homeostasis: the social ori-
gins of interoceptive inference” by Fotopoulou and Tsakiris. Neuropsychoanalysis: An Interdisciplinary Journal for Psychoanal-
ysis and the Neurosciences, 19(1), 29–33. http://doi.org/10.1080/
15294145.2017.1295218. Fraley, R.C., Waller, N.G., Brennan, K.A. (2000). An item response
theory analysis of self-report measures of adult attach-
ment. Journal of Personality and Social Psychology, 78(2), 350–65. http://doi.org/10.1037/0022-3514.78.2.350. Garcia-Larrea, L., Frot, M., Valeriani, M. (2003). Brain gen-
erators of laser-evoked potentials: from dipoles to func-
tional significance. Neurophysiologie Clinique, 33(6), 279–292. http://doi.org/10.1016/j.neucli.2003.10.008. Beckes, L., Coan, J.A. (2011). Social Baseline Theory: the role of
social proximity in emotion and economy of action. Social
and Personality Psychology Compass, 5(12), 976–88. http://doi.org/
10.1111/j.1751-9004.2011.00400.x. Downloaded from https://academic.oup.com/scan/article-abstract/13/11/1121/5106208 by guest on 03 December 2018 Garcia-Larrea, L., Peyron, R. (2013). Pain matrices and neuropathic
pain matrices: a review. Pain, 154, 29–43. http://doi.org/10.1016/
j.pain.2013.09.001. Berscheid, E. (2003). The human’s greatest strength: other
humans. In L. G. Aspinwall & U. M. Staudinger (Eds.) A
Psychology of Human Strengths: Fundamental Questions and
Future Directions for a Positive Psychology, pp. (37–47). Wash-
ington, DC, US: American Psychological Association. http://
doi.org/10.1037/10566-003. Gazzola, V., Spezio, M.L., Etzel, J.A., Castelli, F., Adolphs, R.,
Keysers, C. (2012). Primary somatosensory cortex discrimi-
nates affective significance in social touch. Proceedings of the
National Academy of Sciences, 109(25), E1657–66. http://doi.org/
10.1073/pnas.1113211109. Bowlby, J. (1969). Attachment and Loss, Volume 1: Attachment. New
York: Basic Books. http://doi.org/10.1177/000306518403200125. Gelman, A., Hill, J., Yajima, M. (2012). Why we (usually) don’t have
to worry about multiple comparisons. Journal of Research on
Educational Effectiveness, 5(2), 189–211. Büchel, C., Geuter, S., Sprenger, C., Eippert, F. (2014). Placebo
analgesia: a predictive coding perspective. Neuron, 81(6),
1223–39. Gentsch, A., Panagiotopoulou, E., Fotopoulou, A. (2015). Active
interpersonal touch gives rise to the social softness illusion. Current Biology, 25(18), 2392–7. Coan, J.A., Schaefer, H.S., Davidson, R.J. (2006). Lending a hand
of the neural response to threat. Psychological Science, 17(12),
1032–9. http://doi.org/10.1111/j.1467-9280.2006.01832.x. Geuter, S., Boll, S., Eippert, F., Büchel, C. (2017). Acknowledgements The authors thank Mahmoud El Bahnasawi, Susan Amiri and
Anna Ciaunica for their assistance with data collection. M. von Mohr et al. 1129 M. von Mohr et al. 1129 References pleasant touch. Neuroscience and Biobehavioral Reviews, 34(2),
192–203 http://doi.org/10.1016/j.neubiorev.2009.02.003. References Psychological and neurobi-
ological processes in coping with pain: The role of social
interactions. In: Internation Handbook of Psychobiology. New York:
Routledge. Eisenberger, N.I., Taylor, S.E., Gable, S.L., Hilmert, C.J.,Lieberman,
M.D. (2007). Neural pathways link social support to attenuated
neuroendocrine stress responses. NeuroImage, 35(4), 1601–12. http://doi.org/10.1016/j.neuroimage.2007.01.038. Krahé, C., Paloyelis, Y., Condon, H., Jenkinson, P.M., Williams,
S.C.R., Fotopoulou, A. (2015). Attachment style moderates Essick, G.K., McGlone, F., Dancer, C., Fabricant, D., Ragin, Y.,
Phillips, N., Guest, S. (2010). Quantitative assessment of 1130
Social Cognitive and Affective Neuroscience, 2018, Vol. 13, No. 11 partner presence effects on pain: a laser-evoked potentials
study. Social Cognitive and Affective Neuroscience, 10(8), 1030–7. http://doi.org/10.1093/scan/nsu156. Olausson, H., Lamarre, Y., Backlund, H., et al. (2002). Unmyeli-
nated tactile afferents signal touch and project to insular
cortex. Nature Neuroscience, 5(9), 900–4. http://doi.org/10.1038/
nn896. Krahé, C., Springer, A., Weinman, J. A., Fotopoulou, A. (2013). The social modulation of pain: others as predictive signals of
salience – a systematic review. Frontiers in Human Neuroscience,
7. http://doi.org/10.3389/fnhum.2013.00386. Plaghki, L., Mouraux, A. (2005). EEG and laser stimulation as
tools for pain research. Current Opinion in Investigational Drugs
(London, England: 2000), 6(1), 58–64. Ravitz,
P.,
Maunder,
R.,
Hunter,
J.,
Sthankiya,
B.,
Lancee,
W. (2010). Adult attachment measures: a 25-year review. Journal of Psychosomatic Research, 69(4), 419–432. http://doi. org/10.1016/j.jpsychores.2009.08.006. Lee, M.C., Mouraux, A., Iannetti, G.D. (2009). Characterizing the
cortical activity through which pain emerges from nocicep-
tion. Journal of Neuroscience, 29(24), 7909–16. http://doi.org/
10.1523/JNEUROSCI.0014-09.2009. Sambo,
C.F.,
Howard,
M.,
Kopelman,
M.,
Williams,
S.,
Fotopoulou, A. (2010). Knowing you care: effects of perceived
empathy
and
attachment
style
on
pain
perception. Pain,
151(3),
687–93. http://doi.org/10.1016/j.pain.2010. 08.035. Legrain, V., Iannetti, G.D., Plaghki, L., Mouraux, A. (2011). The pain
matrix reloaded: a salience detection system for the body. Progress in Neurobiology, 93(1), 111–124. http://doi.org/10.1016/
j.pneurobio.2010.10.005. Downloaded from https://academic.oup.com/scan/article-abstract/13/11/1121/5106208 by guest on 03 December 2018 Liljencrantz, J., Strigo, I., Ellingsen, D.M., et al. (2017). Slow brush-
ing reduces heat pain in humans. European Journal of Pain
(United Kingdom), 21(7), 1173–85 http://doi.org/10.1002/ejp.1018. Sharpley, C. F., Rogers, H. J. (1984). Preliminary validation of
the abbreviated Spanier Dyadic Adjustment scale: some psy-
chometric data regarding a screening test of marital adjust-
ment. Educational and Psychological Measurement, 44(4), 1045–9. http://doi.org/10.1177/0013164484444029. Löken, L.S., Wessberg, J., Morrison, I., McGlone, F., Olausson, H. (2009). Coding of pleasant touch by unmyelinated afferents
in humans. Nature Neuroscience, 12(5), 547–8. http://doi.org/
10.1038/nn.2312. References Suvilehto,
J.T.,
Glerean,
E.,
Dunbar,
R.I.M.,
Hari,
R.,
Nummenmaa,
L. (2015). Topography
of
social
touching
depends on emotional bonds between humans. Proceedings of
the National Academy of Sciences, 112(45), 13811–6. http://doi.org/
10.1073/pnas.1519231112. Luck, S.J. (2014). An introduction to the event-related potential tech-
nique, 2nd edn, Cambridge: MIT Press. Mancini, F., Beaumont, A.-L., Hu, L., Haggard, P., Iannetti, G.D.,
Iannetti, G.D.D. (2015). Touch inhibits subcortical and cortical
nociceptive responses. Pain, 156(10), 1936–44 http://doi.org/
10.1097/j.pain.0000000000000253. Tabachnick,
B.G.,
Fidell,
L.S. (2007). Using
Multivariate
Statistics. Pearson Education, Inc., Vol. 28. Boston: Pearson. http://doi.org/10.1037/022267. McGlone, F., Wessberg, J., Olausson, H. (2014). Discriminative and
affective touch: sensing and feeling. Neuron, 82(4), 737–755. Tarkka, I.M., Treede, R.D. (1993). Equivalent electrical source
analysis of pain-related somatosensory evoked potentials
elicited
by
a
CO2
laser. Journal
of
Clinical
Neurophysiol-
ogy, 10(4), 513–519. http://doi.org/10.1097/00004691-199310000-
00009. Mikulincer,
M.,
Shaver,
P.R.,
Pereg,
D. (2003). Attachment
theory and affect regulation: the dynamics, development,
and cognitive consequences of attachment-related strate-
gies 1. Motivation and Emotion, 27(2), 77–102. http://doi.org/
10.1023/A:1024515519160. Triscoli, C., Olausson, H., Sailer, U., Ignell, H., Croy, I. (2013). CT-
optimized skin stroking delivered by hand or robot is compa-
rable. Frontiers in Behavioral Neuroscience, 7, 208. http://doi.org/
10.3389/fnbeh.2013.00208. von Mohr, M., Fotopoulou, A. (2018). The cutaneous borders of
interoception: active and social inference on pain and pleasure
on the skin. In: Tsakiris, M., De Preester, H., editors. The Intero-
ceptive Mind: From Homeostasis to Awareness. Oxford University
Press. Uchino, B.N. (2006). Social support and health: a review of
physiological processes potentially underlying links to dis-
ease outcomes. Journal of Behavioral Medicine, 29 (4), 377–387. http://doi.org/10.1007/s10865-006-9056-5. Morrison, I. (2016). Keep calm and cuddle on: social touch as a
stress buffer. Adaptive Human Behavior and Physiology, 2, 344–62. Valentini, E., Hu, L., Chakrabarti, B., Hu, Y., Aglioti, S. M.,
Iannetti, G.D. (2012). The primary somatosensory cortex
largely contributes to the early part of the cortical response
elicited by nociceptive stimuli. NeuroImage, 59(2), 1571–81. http://doi.org/10.1016/j.neuroimage.2011.08.069. Mouraux, A., Iannetti, G. D. (2009). Nociceptive laser-evoked brain
potentials do not reflect nociceptive-specific neural activ-
ity. Journal of Neurophysiology, 101(6), 3258–69. http://doi.org/
10.1152/jn.91181.2008. Nelson, E.E., Panksepp, J. (1998). Brain substrates of mother-
infant attachment: contributions of opoids, oxytocin and nore-
pinephrine. Neuroscience & Biobehavioral Reviews, 22(3), 437–52. http://doi.org/10.1016/S0149-7634(97)00052-3. Villemure, C., Bushnell, M.C. (2002). Cognitive modulation of
pain: how do attention and emotion influence pain pro-
cessing? Pain, 95(3), 195–199. http://doi.org/10.1016/S0304-3959
(02)00007-6. Olausson, H., Cole, J., Rylander, K., et al. (2008). References Functional
role
of
unmyelinated
tactile
afferents
in
human
hairy
skin:
sympathetic
response
and
perceptual
localization. Experimental
Brain
Research,
184(1),
135–40. http://doi.org/10.1007/s00221-007-1175-x. von
Mohr,
M.,
Kirsch,
L.P.,
Fotopoulou,
A. (2017). The
soothing
function
of
touch:
affective
touch
reduces
feelings of social exclusion. Scientific Reports, 7(1), 13516. http://doi.org/10.1038/s41598-017-13355-7.
|
https://openalex.org/W3102588528
|
https://digital.library.adelaide.edu.au/dspace/bitstream/2440/131869/2/hdl_131869.pdf
|
English
| null |
The Application of Deep Convolutional Neural Networks to Brain Cancer Images: A Survey
|
Journal of personalized medicine
| 2,020
|
cc-by
| 15,795
|
Amin Zadeh Shirazi 1,2
, Eric Fornaciari 3, Mark D. McDonnell 2, Mahdi Yaghoobi 4,
Yesenia Cevallos 5
, Luis Tello-Oquendo 5, Deysi Inca 5 and Guillermo A. Gomez 1,* Amin Zadeh Shirazi 1,2
, Eric Fornaciari 3, Mark D. McDonnell 2, Mahdi Yaghoobi 4,
Yesenia Cevallos 5
, Luis Tello-Oquendo 5, Deysi Inca 5 and Guillermo A. Gomez 1,* 1
Centre for Cancer Biology, SA Pathology and the University of South of Australia, Adelaide, SA 5000,
Australia; amin.zadeh_shirazi@mymail.unisa.edu.au 1
Centre for Cancer Biology, SA Pathology and the University of South of Australia, Adelaide, SA 5000,
Australia; amin.zadeh_shirazi@mymail.unisa.edu.au y
2
Computational Learning Systems Laboratory, UniSA STEM, University of South Australia, Mawson Lakes,
SA 5095, Australia; Mark.McDonnell@unisa.edu.au 2
Computational Learning Systems Laboratory, UniSA STEM, University of South Australia, Mawson Lakes,
SA 5095, Australia; Mark.McDonnell@unisa.edu.au 3
Department of Mathematics of Computation, University of California, Los Angeles (UCLA), Los Angeles,
CA 90095, USA; efornaci@gmail.com 3
Department of Mathematics of Computation, University of California, Los Angeles (UCLA), Los Angeles,
CA 90095, USA; efornaci@gmail.com 4
Electrical and Computer Engineering Department, Islamic Azad University, Mashhad Branch,
Mashad 917794-8564, Iran; yaghoobi@mshdiau.ac.ir 5
College of Engineering, Universidad Nacional de Chimborazo, Riobamba 060150, Ecuador;
jeseniacevallos@hotmail.com (Y.C.); lptelloq@ieee.org (L.T.-O.); deysi_vib@hotmail.es (D.I.) j
p
q
g
*
Correspondence: guillermo.gomez@unisa.edu.au *
Correspondence: guillermo.gomez@unisa.edu.au Received: 29 September 2020; Accepted: 10 November 2020; Published: 12 November 2020 Abstract: In recent years, improved deep learning techniques have been applied to biomedical image
processing for the classification and segmentation of different tumors based on magnetic resonance
imaging (MRI) and histopathological imaging (H&E) clinical information. Deep Convolutional
Neural Networks (DCNNs) architectures include tens to hundreds of processing layers that can
extract multiple levels of features in image-based data, which would be otherwise very difficult
and time-consuming to be recognized and extracted by experts for classification of tumors into
different tumor types, as well as segmentation of tumor images. This article summarizes the latest
studies of deep learning techniques applied to three different kinds of brain cancer medical images
(histology, magnetic resonance, and computed tomography) and highlights current challenges in the
field for the broader applicability of DCNN in personalized brain cancer care by focusing on two
main applications of DCNNs: classification and segmentation of brain cancer tumors images. Keywords: deep learning; DCNN; convolutional neural networks; brain cancer; MRI; histology;
classification; segmentation J. Pers. Med. 2020, 10, 224; doi:10.3390/jpm10040224 Journal of
Personalized
Medicine Journal of
Personalized
Medicine Journal of
Personalized
Medicine Journal of
Personalized
Medicine The Application of Deep Convolutional Neural
Networks to Brain Cancer Images: A Survey Amin Zadeh Shirazi 1,2
, Eric Fornaciari 3, Mark D. McDonnell 2, Mahdi Yaghoobi 4,
Yesenia Cevallos 5
, Luis Tello-Oquendo 5, Deysi Inca 5 and Guillermo A. Gomez 1,* 1. Introduction Artificial Intelligence (AI)and Machine Learning (ML) methods play a critical role in industrial
processes [1–3] and biomedicine [4–7]. By using ML techniques, we can efficiently handle
ambiguous and time-consuming biomedical tasks with nearly the same precision as trained specialists. Advancements in deep learning algorithms as a subfield of ML have demonstrated their strength
in biomedicine data analysis, particularly in cancer data including patients’ images and clinical
information [8–11]. In the recent decade, many researchers have given special attention to Deep Convolutional Neural
Networks (DCNNs) as a very powerful machine-vision tool among deep learning techniques [12]. By applying DCNNs to patients’ X-ray, Computed Tomography (CT), and histopathological images,
various types of cancers such as breast [13,14], prostate [15,16], colorectal [17,18], kidney [19,20],
and brain [21,22] have been diagnosed in their early stages. J. Pers. Med. 2020, 10, 224; doi:10.3390/jpm10040224 www.mdpi.com/journal/jpm J. Pers. Med. 2020, 10, 224 2 of 27 Brain cancer is the first leading cause of mortality among females age 20 and younger, and males age
40 and younger [23]. A large collection of brain cancer patients’ images have been curated and are now
rapidly available [24]. Studies show that brain tumors are highly heterogeneous [25] which constitute
the main problem for brain tumor classification and segmentation and therefore diagnosis and prognosis. Recently, a very high-quality prospective survey has been done by Muhammad et al. [26] to classify
multigrade brain tumors based on Magnetic Resonance (MR) images. In this study, they explored the
impact of primary stages such as preprocessing, data augmentation, transfer learning, and different
Convolution Neural Network (CNN) architectures on the CNN-based classifiers performance. After some inspections, we realized that there is a gap in the application of DCNNs to classify or
segment images of brain cancer and most of studies/surveys have just focused on MR or CT images
while there is highly-valuable information in the background of histopathological imaging (H&E)
images. Therefore, in this literature review, we explore recent advances published between 2018 to 2020
in the use of supervised DCNNs as a robust machine-vision tool in the classification and segmentation
tasks of three different kinds of brain cancer images including H&E histology, MR, and CT. For this,
articles were searched using the following keywords: “classification”, “segmentation”, “detection”,
“brain cancer”, “brain tumor”, “brain tumor lesion”, “brain malignancy”, “brain malignant tissue”,
“CNN”, “DCNN”, “convolutional neural network”, “histology”, “pathology”, “histopathology”,
“Magnetic Resonance Imaging (MRI)”, “CT”, “imaging”, and “image”. 1. Introduction We then assessed the list of
identified articles for its content and contribution to the field, in order to include these in this survey. Finally, we also included in this survey some older articles, whose content have contributed key
advances in this field. All the works discussed in this survey have used supervised learning to train
deep convolutional neural networks. From the selected studies in the classification and segmentation parts, we found that 32% and 40%
of studies have been supervised/validated by a specialist (e.g., pathologists/radiologists), respectively. In these studies, a specialist has been involved in i) dataset preparation or ii) result validation obtained
by the methods used in the classification and segmentation parts (for further supervision). In the rest
of the articles, the results have been evaluated by testing samples whose results were compared with
the ground truths (labels/masks) created by specialists. Our focus in this work is to provide a legible and concise explanatory paragraph for each recent
and relevant work. For this purpose, we did our best to describe the main points in each article in a
way that all readers (even the readers who are not very familiar with convolutional neural networks)
are able to familiarize with the recent advances in this field. For simplicity, all information provided
related to the classification and segmentation tasks have been summarized into two tables. We have
also depicted the most important and influential steps of these two tasks in two separated figures as a
roadmap for researchers who are going to apply DCNNs in their future work for the classification or
segmentation of different kinds of brain cancer images. Furthermore, for more clarity, the articles in each section were sorted based on the complexity of
their methods/algorithms used from simple to complex. The rest of this article is organized as follows: In Sections 2 and 3, novel articles applying DCNNs
for classification and segmentation tasks to brain cancer images are explored. Indeed, for simplicity,
all information for the two tasks mentioned are briefly compiled in two tables. We have also
summarized the primary steps and proposed a roadmap for those tasks into separate figures. Finally,
in Sections 4 and 5, we discuss the main contributions of the reviewed works, current limitations and a
conclusion summarizing future directions for this rapidly developing field. 2. DCNNs Application in the Classification of Brain Cancer Images In this section, the application of DCNNs to classify brain cancers from different kinds of brain
cancer images is explained. For this purpose and to add clarity, we have divided this part into three
subsections with each of these focusing on a specific type of images (i.e., H&E histology, MRI, or both). J. Pers. Med. 2020, 10, 224 3 of 27 2.1. DCNNs Application in the Classification of Brain Cancer H&E Histology Images 2.1. DCNNs Application in the Classification of Brain Cancer H&E Histology Images Zadeh Shirazi et al. [24] proposed a new DCNN-based classifier, namely DeepSurvNet, short for
“Deep Survival Convolutional Network”, to accurately classify the survival rates of brain cancer
patients. The model assigns ranges of survival likelihood across four classifications based on H&E
histopathological images. The datasets are collected from the TCGA and a local public hospital,
including 450 patients’ H&E slides with different kinds of brain cancer tumors. They considered classes
I, II, III, and IV for patients with 0–6, 6–12, 12–24, and more than 24 months of survival after diagnosis,
respectively. DeepSurvNet is based on the GoogLeNet [27] architecture and was trained and tested on
a public brain cancer dataset from TCGA, and also generalized on a new private dataset. Their model
achieved precisions of 0.99 and 0.8 (recalls of 0.98 and 0.81) in the testing stages on the two datasets,
respectively. Furthermore, they analyzed the frequency of mutated genes associated with each class,
supporting the idea of a different genetic fingerprint associated with patient survival. Sidong et al. [28] focused on the Isocitrate Dehydrogenase (IDH), an important biomarker in
glioma, and predicted its mutational status by using DCNN. Their dataset includes 266 H&E slides
gliomas of grade 2 to 4 collected from TCGA and a private hospital. They proposed a model based
on using Generative Adversarial Networks (GAN) methodology to generate synthetic but realistic
samples to support data augmentation and Resnet50 DCNN architecture as a primary backbone for
IDH status prediction. They also concluded that by adding patients’ age as a new feature, the DCNN
model can predict IDH status more accurately. They achieved an accuracy of the IDH mutational status
of 0.853 (Area Under the Curve (AUC) = 0.927). Sumi et al. 2. DCNNs Application in the Classification of Brain Cancer Images [29] put forward an architecture which can classify four different types of brain tissues
of categories healthy, benign, Oligodendroglioma, and Glioblastoma (GBM) based on features extracted
from the cellular level (not the tissue level). Their model inputs are H&E histology images of brain
tumors. Their model is based on a spatial fusion network where an entire image-wise prediction can
discriminate between different sparse features in a whole image. To achieve this, a preprocessing stage
extracts patches from each image and applies augmentation. Next, the InceptionResNetv2 (INRV2)
architecture is trained on the augmented patches and predicts probabilistic features of different kinds
of tumor cancers for local patches. In the second stage, a deep spatial fusion network is implemented,
which includes several Fully Connected (FC) and dropout layers to learn spatial relationships between
local patches. Finally, a vector calculates the patch tumor class (patch-wise probability). In their work,
two datasets, The Cancer Genome Atlas (TCGA) and The Cancer Imaging Archive (TCIA) with 2034
and 2005 basic images have been used, though the training set could be increased with additional data
augmentation methods. Additionally, the total patches were extracted, equal to 202,174, and 140,099
from the TCGA and TCIA datasets, respectively. Their model achieved classification accuracy #1 of
0.95 on four-class classification and the classification accuracy #2 of 0.99 on two-class classification
(non-necrosis and necrosis). Yonekura et al. [30] proposed a 14-layer DCNN-based architecture to classify GBM and Low-Grade
Glioma (LGG) images. The dataset consists of 200 H&E histological Whole Slide Images (WSIs) from
TCGA which contain 100 images. 10,000 distinct patches are extracted from each cohort as inputs to
train the DCNN. Additionally, for further performance checking, some popular DCNN architecture
such as LeNet [31], ZFNet [32], and VGGNet [33] were trained on those patches and their results
were compared with the proposed model. Finally, a classification accuracy of 0.96 for four-fold
cross-validation was achieved by the model. 2.2. DCNNs Application in the Classification of Brain Cancer MR Images Fukuma et al. [34] implemented a novel architecture combining several sets of features to
identify IDH mutations and TERT promoter (pTERT) mutations. These molecular characteristics are
crucial indicators for diagnosing and treating gliomas grades II/III. The model combines patient age,
61 conventional radiomic features, 3 tumor location parameters, and 4000 texture features as input. The texture features are extracted from normalized and cropped tumor lesion slides using AlexNet. J. Pers. Med. 2020, 10, 224 4 of 27 Of these feature sets, a subset is selected using F-statistics. To increase the total dataset, augmentation
is used. The final accuracy on a nonpublic dataset is 63.1%, which outperforms previous models using
only single feature sets. y
g
Chang et al. [35] applied the ResNet50 deep learning model to noninvasively predict IDH1 and
IDH2 mutations in glioma grades II-IV. The initial dataset is built from several medical institutions:
Hospital of the University of Pennsylvania, Brigham and Women’s Hospital, The Cancer Imaging
Center, Dana-Farber/Brigham and Women’s Cancer Center. Skull-stripping, intensity normalization,
and tumor extraction preprocessing are applied to the MRI images. The resulting dataset was input
into a single combined ResNet50 network that resulted in test accuracies of 87.6%. Additional fields
such as age have been shown to improve accuracy by to 89.1%. Alqudah et al. [36] put forth a new DCNN architecture with just 18 layers for classifying
(grading) MR images into three classes of tumors including Meningioma, Glioma, and Pituitary. They used a public dataset containing 3064 brain MR images (T1 weighted contrast-enhanced). In the
preprocessing stage, all images are converted into three distinct categories including cropped lesions,
uncropped lesions, and segmented lesions. Then, they trained their own DCNN on these three
categories and found out that the highest overall performance is related to the uncropped lesions
category with 99% accuracy and 98.52% sensitivity. Kalaiselvi et al. [37] designed and implemented six different DCNN-based classifiers to distinguish
LGG and High-Grade Glioma (HGG) tumors from normal lesions through brain cancer MR images. Each model contains just two to five layers. First, each model is trained on the BraTS2013 dataset with
4500 images. The criterion used as a hyperparameter to adjust the models and prevent them from
overfitting is early stopping. Then, in the testing phase, they utilized the The Whole-Brain Atlas (WBA)
dataset with 281 images. 2.2. DCNNs Application in the Classification of Brain Cancer MR Images 2020, 10, 224 layers were applied to feature maps much closer to the network input. The output of these layers was
concatenated to the output global average pooling layer, in order to fuse high and low-level features. layers were applied to feature maps much closer to the network input. The output of these layers was
concatenated to the output global average pooling layer, in order to fuse high and low-level features. They call their network G-ResNet which is short for Global Average Pooling Residual Network. In their
experiments, they tested four combinations of concatenation layers between different layers to integrate
low-level and high-level features and determine which resulted in the best model accuracy. Indeed,
they realized modifying loss function or using the sum of the loss functions such as cross-entropy and
interval, the model accuracy would improve. Based on all modifications they applied, their model
accuracy could achieve 0.95 as total classification accuracy. Hemanth et al. [40] proposed a simple DCNN architecture with some modifications in the FC layer. This proposed model considerably reduced computational complexity and is able to classify four brain
tumor types of Meningioma, Glioma, Metastasis, and Astrocytoma. T1, T2, and T2 flair MR images
are fed as inputs into the model. A total of 220 images were used in their paper, as collected from a
private clinic. Their main contribution is to eliminate the updating procedure of weights by using the
GD algorithm in the FC layers by replacing them with a simplified approach such that the number of
trainable parameters is greatly reduced. They proved that with this amendment, the proposed model
can classify brain tumors with an accuracy of 0.96. Afshar et al. [41] developed a new classifier based on CapsNets to classify three types of brain
cancer tumors. To train their proposed network, they used a publicly available dataset including 3064
T1-weighted MR images related to Meningioma, Glioma, and Pituitary tumors. Although CapsNets
have some benefits over traditional DCNNs, they are considerably more sensitive to image backgrounds. Hence, preprocessing stages such as tumor segmentation from the whole image and removing
backgrounds are highly recommended. 2.2. DCNNs Application in the Classification of Brain Cancer MR Images However, in this work, the authors reinforced the CapsNets
architecture by providing the whole tumor images as the main inputs of their model and the tumor
surrounding tissues as extra inputs in its final layer as they considered that tumor segmentation
procedure can at the same time introduce disadvantages as it is not only a time-consuming task but also
can eliminate some important information such as the tumor boundaries. As a result, their proposed
classifier accuracy of 0.91 shows that their developed CapsNet outperforms the previous CapsNets. Seetha et al. [42] used a transfer learning approach in their DCNN architecture to distinguish brain
tumors from nontumors. They applied a DCNN model pretrained on the ImageNet dataset. The public
datasets used in their work are Radiopaedia and BraTS2015 containing brain MR images. In their
approach, all layers in the pretrained model are frozen, excluding the final layer. The training procedure
on the new MR images remains the same in the latest layer. Although this type of classification task
only differentiates two classes from each other (i.e., tumor and normal tissues), the model was able to
save substantial time in the training phase by using transfer learning method and ultimately achieved
a classification accuracy of 0.97. Pashaei et al. [43] proposed an ensemble model Kernel Extreme Learning Machine-CNN (KE-CNN)
combining DCNN and the Kernel Extreme Learning Machine (KELM). In their simple model of just
9 layers, the DCNN extracts features from input images and are fed as input vectors to the KELM
as the final layer. In other words, KELM is used as an alternative to FC layers and is responsible for
classifying the images. They used this model to classify 3064 T1-weighted MR images from a public
dataset. This dataset contains three different kinds of brain tumors including Meningioma, Glioma,
and Pituitary. Based on their results, the highest classification accuracy of 0.93 was achieved when
they applied the radial base function as a kernel function in the KELM classifier. Zhou et al. [44] put forward a classifier based on DenseNet and LSTM (DenseNet-LSTM). In this
model, features are extracted from MR images with an autoencoder architecture i.e., DenseNet. Those features are then fed to the LSTM structure as inputs for classification. They applied these
methods to two distinct public and private datasets for training and evaluating the proposed model. 2.2. DCNNs Application in the Classification of Brain Cancer MR Images After the training and testing stages, they realized that among the six models,
the Five-Layer Model with Stopping Criteria and Batch Normalization (FLSCBN) achieved the lowest
3% False Alarm (FA) and 7% Missed Alaram (MA) (as error rates indexes) and a highest accuracy
classification of 89%. Mzoughi et al. [22] applied a DCNN with 11 layers to 3D whole MR images to classify the grade
of glioma tumors into LGG or HGG. In their approach, whole 3D volumetric MRI sequences are
passed to the DCNN instead of patch extraction from the MR image. In this study, the dataset used is
BraTS2018, which comprises 351 MR T1-weighted images, including mixed grades of glioma tumors. To solve the problem of data image heterogeneity, all images are preprocessed using adaptive contrast
enhancement and intensity normalization methods. Furthermore, to generate additional training
data, augmentation was used. Based on the results, their proposed 3D DCNN model achieved an
overall accuracy of 0.96 on validation data images, which is relatively higher in comparison with other
traditional and shallow DCNN approaches. Badža et al. [38] presented a relatively simple DCNN-based architecture with only 22 layers to
classify three brain cancer tumors including Meningioma, Glioma, and Pituitary. The model was trained
on a public dataset which contains 3064 T1-weighted contrast-enhanced MR images. With regard
to their model inputs being MR whole images than extracted patches, they increased the number of
images three times, i.e., to 9192, by using data augmentation techniques (images vertical flipping and
90-degree rotation). The proposed architecture was tested several times and finally found out the
best result with the accuracy of 96.56% is related to the conditions that the model is validated by the
augmented dataset and 10-fold cross-validation method. Their simple model would be suitable for
users who are going to run the network with limited hardware, such as mobile phones or conventional
PCs and who want to see the results quickly. Liu et al. [39] designed and implemented a model to classify three distinct types of brain tumors
including Meningioma, Glioma, and Pituitary. The publicly available dataset used in their network is
of 3064 T1-weighted contrast-enhanced MR images. Their model is powered by ResNet34 architecture,
which uses an all-convolutional architecture with a global pooling layer right before the output. They modified the standard ResNet architecture slightly so that additional global average poling 5 of 27 J. Pers. Med. 2.2. DCNNs Application in the Classification of Brain Cancer MR Images The public dataset includes 3064 brain cancer MR images and three types of tumors containing Gliomas,
Pituitary, and Meningiomas. In contrast, the brain tumor types in the private dataset are completely
different including Glioma, Meningiomas, Metastatic tumors and normal lesions. This dataset contains J. Pers. Med. 2020, 10, 224 6 of 27 422 MR images. The DenseNet-LSTM was trained and tested on both datasets separately and achieved
0.92 and 0.71 of classification accuracies, respectively. Mohsen et al. [45] used a DNN with seven layers as their classifier rather than DCNN because
of their limited hardware resources. In their proposed model, first, brain tumors/normal lesions
are segmented by using the Fuzzy C-means method. Next, the DWT technique extracts the major
features from the segmented brain tumor MR images and the features are reduced using the Principal
Component Analysis (PCA) method. The extracted features are fed into the next part as the inputs of
the classifier. Finally, a DNN architecture is applied to classify three different types of brain tumors
including GBM, Sarcoma, and Metastasis along with normal brain lesions. The dataset used in this
work is publicly available and comprises 66 T2 weighted brain MR images. Although their proposed
model is not using DCNN for feature extraction and classification, the model achieved a classification
accuracy and AUC of 0.97 and 0.98, respectively. Ari et al. [46] proposed different models to classify brain cancer as benign or malignant brain cancer
tumors. The dataset used in their work contains 16 patients who have been diagnosed with a brain
tumor via T1-weighted MR images screening. First, the images are preprocessed and the background
noise is filtered. In the next stage, two different kinds of classifiers such as a six-layer DCNN and
ELM local receptive fields (ELM-LRF) were used to classify the tumor types. The results show that the
two classifiers have roughly the same classification accuracy of 0.96 and 0.97, respectively where the
ELM-LRF achieved slightly better performance. However, the lower accuracy of the DCNN model
could be attributed to the limited number of images in the training part. Suter et al. [47] put forth two different DCNN-based approaches to predict survival rate in GBM
patients. Both used the BraTS2018 dataset containing 293 T1, T1c, T2, and T2-Weighted Fluid-Attenuated
Inversion Recovery (FLAIR) MR images. To increase the number of images in the training phase,
the images were first segmented. 2.2. DCNNs Application in the Classification of Brain Cancer MR Images DCNNs Application in the Classification of Brain Cancer H&E Histology and MR Images
g
p
g
g
g
images and subsequently detect/classify/predict relevant diseases are extremely
li i i
d
i
i
h d i
l di
CNN b
d l
ifi
d
hi
k 2.3. DCNNs Application in the Classification of Brain Cancer H&E Histology and MR Images
images and subsequently detect/classify/predict relevant diseases are extremely e
li i i
d
i
i
th d i
l di
CNN b
d l
ifi
d
thi t
k Bagari et al. [50] put forward a novel DCCN approach by using both H&E WSIs and MR images to
classify low-grade gliomas into categories of Astrocytoma and Oligodendroglioma. The dataset used in
this work contains 30 and 20 different patients in the training and testing phase, respectively. All patients
have both MR and H&E histological images. First, preprocessing stages such as RoI extraction from
H&E histological images, patch extraction, and stain normalization were used. Then, an autoencoder
detected abnormal patches among all patches, and finally, a DenseNet-based architecture was applied
to the abnormal patches to distinguish Astrocytoma tissues from Oligodendroglioma tissues. Next,
a trained 3D DCNN model was applied on MR images including FLAIR, T1, T1C, and T2 MR sequences
to segment the tumor lesions. Then, another DCNN architecture extracts the principal features from
segmented tumors and feeds them as inputs to a logistic linear regression classifier. Finally, the classifier
detects the type of low-grade glioma tumors and combines the results with the previous classifier
and results in the total performance of the model. Their combined approach achieved a classification
accuracy of 0.9. clinicians, and noninvasive methods including CNN-based classifiers can do this task very well. In
general, CNN-based classifiers might be applied to three different kinds of brain cancer images
including H&E histology, MR, or both. These classifiers can categorize various brain tumor types or
predict biomarker status and patients’ survival rate based on the features inside the images. Hence,
two separated and important parts can be considered to achieve the best results: preprocess and CNN
architecture. Based on the various studies that we completed, some steps which play important roles
in the preprocessing part include patch extraction, abnormal patch detection/selection, intensity
normalization, contrast enhancement, and background removing. 2.2. DCNNs Application in the Classification of Brain Cancer MR Images Overall, there is no best practice to
recommend one or several steps in the preprocessing phase and researchers have to try many
experiments to find the best results. However, studies show that data augmentation techniques to
expand the number of data samples as model inputs have successfully helped the CNNs to achieve
higher accuracy. In the second part, CNN models can be first trained to extract features from brain
cancer images, and then, those feature vectors along with patients’ clinical data together make a
numerical dataset
Afterwards
some linear regression models or nonlinear machine learning y
All information related to the classification/prediction task mentioned above is summarized in
Table 1. We have also depicted in Figure 1 the main steps of this part as a roadmap for researchers who
are going to apply DCNNs in their future work for the classification task. numerical dataset. Afterwards, some linear regression models or nonlinear machine learning
methods use this dataset to classify/predict relevant biomarker status, survival rate, or brain tumor
types. However, It would be simpler to directly apply CNNs as a classifier or predictive model
(Figure 1). Figure 1. Shows roadmap and stages of a typical brain tumor classification architecture with the
following high-level steps: 1. Input: Haematoxylin and eosin (H&E) stained histology or magnetic
resonance imaging (MRI) can be considered as inputs into the model; 2. Preprocessing: apply several
techniques to remove background, normalize images, patch extraction and data augmentation; Step 3:
Deep Convolutional Neural Network (DCNN) Application: The preprocessed dataset is fed into a DCNN
model. The model can be used a feature extractor or classify/predict the outputs, directly; if the DCNN
model is applied as a feature extractor, these features can be combined with patients clinical information
and make a numerical dataset to apply as inputs of machine learning models to classify/predict
the outputs (the architecture is now outdated, but is used because its relevant for cited papers);
4. Model outputs: brain cancer biomarker status prediction, tumor types classification, or survival rate
classification/prediction (Pr). Figure 1. Shows roadmap and stages of a typical brain tumor classification architecture with the
following high-level steps: 1. Input: Haematoxylin and eosin (H&E) stained histology or magnetic
resonance imaging (MRI) can be considered as inputs into the model; 2. 2.2. DCNNs Application in the Classification of Brain Cancer MR Images Then both the raw and segmented images were fed into the model as
inputs. In the first model, a simple DCNN with five blocks was used. In the second DCNN architecture,
clinical information such as patients’ age and resection status was added as extra features into the final
FC layer of the model. However, the models’ performance results were not satisfactory. When the
extracted features from the second DCNN model (as deep features) were combined with additional
clinical information such as tumors intensity, location, and shape, and an Support Vector Classifier
(SVC) was used as a final predictive model it led to better prediction results. Their final model was
called Ensemble SVC and achieved a survival rate prediction with 0.42 accuracy on the test samples. Banerjee et al. [48] proposed three distinct DCNN architectures trained on MR images to distinguish
between HGG and LGG. These models are trained on extracted patches from MRIs, simple slices,
and 3D volumetric multiplanar slices and called PatchNet, SliceNet, and VolumeNet, respectively. To maintain a high number of samples in the training phase and assess all proposed models in the
testing phase, the scheme leave-one-patient-out was used. The dataset used in this work includes
491 T1, T1c, T2, and FLAIR MR images of HGG and LGG samples collected from the TCIA public data
centre. PatchNet is shallower than the other two models and because of the bigger size and complexity
of input images in SliceNet and VolumeNet, they are deeper. The results indicate that the deepest
DCNN architecture i.e., VolumeNet trained on the 3D volumetric dataset achieves better classification
results with a classification accuracy of 0.97 on the test dataset. Afshar et al. [49] designed and implemented a classifier based on CapsNets to classify three types
of brain cancer tumors. The proposed network used a dataset including 3064 T1-weighted MR images
related to Meningioma, Glioma, and Pituitary tumors. The CapsNets outperformed traditional DCNNs
considering they can better handle overfitting problems caused by insufficient training instances. In this work, the proposed CapsNet was applied to brain segmented regions and brain whole images as
their model inputs. Eventually, the CapsNet was determined to perform better on segmented tumors
with a classification accuracy of 0.86. J. Pers. Med. 2020, 10, 224
Table 1. We have also
who are going to app 7 of 27
archers 2.3. 2.2. DCNNs Application in the Classification of Brain Cancer MR Images Preprocessing: apply several
techniques to remove background, normalize images, patch extraction and data augmentation; Step 3:
Deep Convolutional Neural Network (DCNN) Application: The preprocessed dataset is fed into a DCNN
model. The model can be used a feature extractor or classify/predict the outputs, directly; if the DCNN
model is applied as a feature extractor, these features can be combined with patients clinical information
and make a numerical dataset to apply as inputs of machine learning models to classify/predict
the outputs (the architecture is now outdated, but is used because its relevant for cited papers);
4. Model outputs: brain cancer biomarker status prediction, tumor types classification, or survival rate
classification/prediction (Pr). 8 of 27 J. Pers. Med. 2020, 10, 224 Table 1. DCNN-based classifiers brief description (sorted by year published/DCNN performance). Ref. Year
Task
Tumor Type
Image Type
Model Name
Model Desc. Software
Hardware
Dataset
Instances/
Cases
Perf. 2.2. DCNNs Application in the Classification of Brain Cancer MR Images [26] **
2020
Classification
–
Glioma
–
Meningioma
–
Pituitary
–
Glioblastoma
MRI
Six DCNN
Architectures *
A prospective
survey on Deep
Learning
techniques applied
for Multigrade
Brain Tumor
Classification
–
Caffe
–
NVidia DIGITS
NVidia TITAN X
(Pascal)
–
multigrade brain
tumor [51]
–
Brain tumor public
data Set [52]
–
TCIA [53]
–
BraTS 2015 [54]
–
Harvard
whole-brain Atlas
[55]
–
Internet Brain
Segmentation
Repository [56]
–
121
–
3064
–
49
–
274
–
30
–
18
–
Accuracy: 0.93
(Achieved by
VGGNet [33] on
multigrade brain
tumor [51])
–
Accuracy: 0.94
(Achieved by
VGGNet [33] on
brain tumor public
data [52])
[24] **
2020
Classification
–
Astrocytoma
–
Mixed-glioma
–
Oligodendroglioma
–
Glioblastoma
H&E Histology
DeepSurvNet
Brain cancer
patients’ survival
rate classification
by using deep
convolutional
neural network
–
Python
–
TensorFlow
–
Keras
4xNVidia 1080 Ti
GPU
–
TCGA [57]
–
Private dataset
–
400
–
9
–
Precision: 0.99
–
Precision: 0.80
[28] **
2020
Prediction
–
GBM
–
LGG
–
Gliomas (Grade
II to IV)
H&E Histology
GAN-based
ResNet50 *
Gliomas’ IDH
status prediction
by using the GAN
model for data
augmentation and
Resnet50 as a
predictive model
–
Python
–
TensorFlow
N/A
–
TCGA [57]
–
Privatedataset
–
200
–
66
–
Accuracy: 0.88
–
AUC: 0.93
[36]
2020
Classification
–
Glioma
–
Meningioma
–
Pituitary
MR (T1 weighted
contrast-enhanced)
18 layers
DCNN *
Meningioma,
glioma, and
pituitary tumors
classification by
using 18 layers
DCNN-based
model on MR
images
N/A
–
Intel
Core-I7 processor
–
16 Gb RAM
Brain tumor public
dataset [58]
3064
–
Accuracy: 0.99
–
Sensitivity: 0.98
[38]
2020
Classification
–
Glioma
–
Meningioma
–
Pituitary
MR (T1 weighted
contrast-enhanced)
22 layers
DCNN *
Meningioma,
glioma, and
pituitary tumors
classification by
using 22 layers
DCNN-based
model based on
MR images
MATLAB R2018a
NVidia 1050 Ti GPU
Brain tumor public
dataset [58]
3064
Accuracy: 0.96 Table 1. DCNN-based classifiers brief description (sorted by year published/DCNN performance). J. Pers. Med. 2020, 10, 224 9 of 27 Table 1. Cont. Ref. Year
Task
Tumor Type
Image Type
Model Name
Model Desc. Software
Hardware
Dataset
Instances/
Cases
Perf. 2.2. DCNNs Application in the Classification of Brain Cancer MR Images [37] **
2020
Classification
–
LGG
–
HGG
MR (T2 weighted)
FLSCBN
Tumor vs
non-tumor
classification by
using a five layers
DCNN-based
model on MR
images
–
Python
–
TensorFlow
–
Keras
–
Intel
Core-I5 processor
–
4GB RAM
–
BraTS2013 [59]
–
WBA [60]
–
4500
–
281
–
Accuracy: 0.89
–
FA: 0.3
–
MA: 0.7
[22] **
2020
Classification
–
LGG
–
HGG
MR (T1-Gado or
T1-weighted)
3-D DCNN
11 layers 3-D
DCNN-based
model to classify
glioma tumors
into LGG and
HGG using the
T1-weighted MR
images
–
Python
–
TensorFlow
–
Keras
–
Intel
Core-I7 processor
–
19.5 GB RAM
–
NVIDI1080
Ti GPU
BraTS2018 [61]
351
Accuracy: 0.96
[34] **
2019
Classification
Glioma
MRI
AlexNet;
Linear Support
Vector Machine
Identify IDH and
pTERT mutations
using age,
radiomic features,
and tumor texture
features
Caffe
N/A
Not Publicly Available
164
Accuracy: 63.1%
[29]
2019
Classification
–
Normal
–
Benign
–
Oligodendroglioma
–
GBM
H&E Histology
INRV2-based
deep spatial
fusion network
*
A mixed DCNN
architecture
combining
InceptionResNetV2
and deep spatial
fusion network to
classify four
different kinds of
brain tumors
based on H&E
images
PyTorch
NVidia 1080 Ti GPU
TCGA [57]
TCIA [53]
–
2034
–
2005
–
Accuracy#1: 0.95
–
Accuracy#2: 0.99
[39]
2019
Classification
–
Glioma
–
Meningioma
–
Pituitary
MR (T1 weighted
contrast-enhanced)
G-ResNet
Meningioma,
glioma, and
pituitary tumors
classification by
using a
ResNet34-based
model with global
average pooling
and modified loss
function based on
MR images
PyTorch
NVidia 1080 Ti GPU
Brain tumor public
dataset [58]
3064
Accuracy: 0.95 J. Pers. Med. 2020, 10, 224 10 of 27 Table 1. Cont. Table 1. Cont. Ref. Year
Task
Tumor Type
Image Type
Model Name
Model Desc. Software
Hardware
Dataset
Instances/
Cases
Perf. 2.2. DCNNs Application in the Classification of Brain Cancer MR Images [40]
2019
Classification
–
Metastasis
–
Meningioma
–
Glioma
–
Astrocytoma
MR (T1, T2, and T2
flair)
MDCNN
Metastasis,
Meningioma,
Glioma and
Astrocytoma
tumors
classification using
a modified DCNN
with reduced
computational
complexity based
on MR images
N/A
N/A
Brain tumor private
dataset [62]
220
Accuracy: 0.96
[30]
2018
Classification
–
GBM
–
LGG
H&E Histology
Deep CNN
GBM and LGG
classification by
using
DCNN-based
model based on
H&E Histological
images
–
Python
–
TensorFlow
–
NVidia 1080
Ti GPU
–
Intel
Core-I7 processor
–
32 GB RAM
TCGA [57]
200
Accuracy: 0.96
[50]
2018
Classification
–
Astrocytoma
–
Oligodendroglioma
H&E Histology;
MR FLAIR, T1, T1C,
and T2 images
A combined
DCNNs-based
network *
Astrocytoma and
Oligodendroglioma
classification by
using
DCNN-based
model based on
both MR and
Histological
images
N/A
N/A
Private dataset
50
Accuracy: 0.90
[43]
2018
Classification
–
Glioma
–
Meningioma
–
Pituitary
MR (T1 weighted
contrast-enhanced)
KE-CNN
Meningioma,
glioma, and
pituitary tumors
classification by
using a mixed
approach of
DCNN and
extreme learning
based on MR
images
N/A
N/A
Brain tumor public
dataset [58]
3064
Accuracy: 0.93 11 of 27 J. Pers. Med. 2020, 10, 224 Table 1. Cont. Table 1. Cont. Ref. Year
Task
Tumor Type
Image Type
Model Name
Model Desc. Software
Hardware
Dataset
Instances/
Cases
Perf. [44]
2018
Classification
Public dataset:
–
Glioma
–
Meningioma
–
Pituitary
–
Private dataset:
–
Normal
–
Meningioma
–
Glioma
–
Metastasis
–
MR (T1 weighted
contrast-enhanced);
–
MR FLAIR, T1,
T1C, and
T2 images
DenseNet-LSTM
–
Meningioma,
glioma, and
pituitary
tumors
classification
by using
DenseNet-LSTM
based on
MR images
–
Normal
lesion and
Meningioma,
Glioma, and
Metastasis
tumors
classification
by using
DenseNet-LSTM
based on
MR images
–
Python
–
TensorFlow
Nvidia Titan Xp
GPU
–
Public dataset [58]
–
Private dataset
–
3064
–
422
–
Accuracy: 0.92
–
Accuracy: 0.71
[41]
2018
Classification
–
Glioma
–
Meningioma
–
Pituitary
MR (T1 weighted
contrast-enhanced)
CapsNet
Meningioma,
Glioma, and
Pituitary tumors
classification by
using a developed
CapsNet
architecture based
on MR images
–
Python
–
Keras
N/A
Brain tumor public
dataset [58]
3064
Accuracy: 0.91
[49]
2018
Classification
–
Glioma
–
Meningioma
–
Pituitary
MR (T1 weighted
contrast-enhanced)
CapsNet
Meningioma,
Glioma, and
Pituitary tumors
classification by
using a developed
CapsNet
architecture based
on MR images
N/A
N/A
Brain tumor public
dataset [58]
3064
Accuracy: 0.86
[42]
2018
Classification
–
Tumor (N/A)
–
Non-Tumor
(normal)
MR
Pre-trained
DCNN
Brian tumors vs
nontumors
classification by
using a pretrained
DCNN based on
MR images
Python
N/A
–
Radiopaedia [63]
–
BraTS2015 [64]
N/A
Accuracy: 0.97 J. Pers. Med. 2020, 10, 224 12 of 27 Table 1. Cont. Table 1. Cont. Ref. Year
Task
Tumor Type
Image Type
Model Name
Model Desc. Software
Hardware
Dataset
Instances/
Cases
Perf. Table 1. Cont. [45]
2018
Classification)
–
Normal
–
GBM
–
Sarcoma
–
Metastasis
MR (T2 weighted)
DWT-DNN *
Normal lesion and
GBM, Sarcoma,
and Metastasis
tumors
classification by
using DWT-DNN
based on MR
images
–
MATLAB R2015a
–
WEKA 3.9
N/A
Public dataset [65]
66
–
Precision: 0.97
–
AUC: 0.98
[46]
2018
Classification
–
Benign
–
Malignant
MR (T1 weighted)
DCNN vs
ELM-LRF *
Benign vs
malignant tumors
classification by
using DCNN and
ELM-LRF models
on MR images
MATLAB R2015a
N/A
Public dataset [66]
16
–
Accuracy: 0.96
–
Accuracy: 0.97
[47] **
2018
Prediction
GBM
MR (T1-weighted, T1c,
T2-weighted, FLAIR)
SVC Ensemble
GBM patient
survival rate
classification by
using two different
DCNN models
based on MR
images
Python
Nvidia Titan Xp
GPU
BraTS2018 [61]
293
Accuracy: 0.42
[48]
2018
Classification
–
GBM
–
LGG
MR (T1-weighted, T1c,
T2-weighted, FLAIR)
–
PatchNet
–
SliceNet
–
VolumeNet
GBM and LGG
classification by
using
DCNN-based
models based on
MR images
–
Python
–
TensorFlow
–
Keras
–
NVidia
1080 GPU
–
Intel
Core-I7 processor
–
32 GB RAM
–
TCGA-GBM [67]
–
TCGA-LGG [68]
–
TCIA [53]
461
Accuracy: 0.97
[35] **
2018
Classification
Glioma
MRI
ResNet50
Identify IDH1/2
mutations in
glioma grades
II-IV using
ResNet50
–
Kera
–
Tensorflow
N/A
From Hospital of the
University of
Pennsylvania, Brigham
and Women’s Hospital,
The Cancer Imaging
Center,
Dana-Farber/Brigham
and Women’s Cancer
Center
603, 414, 471
(With respect
to sources)
Accuracy: 85.7%
* Model names with asterisks are not defined in the original papers and names were assigned based on the models applied. Note: for abbreviations description in this table please refer to
the list of abbreviations on the back partof this article (before References). ** the references with “**” mean that the results achieved by their methods or the dataset used have been
validated/supervised by specialists (e.g., pathologists/radiologist). * Model names with asterisks are not defined in the original papers and names were assigned based on the models applied. Note: for abbreviations description in this table please refer t
the list of abbreviations on the back partof this article (before References). ** the references with “**” mean that the results achieved by their methods or the dataset used have bee
validated/supervised by specialists (e.g., pathologists/radiologist). J. Pers. Med. Table 1. Cont. 2020, 10, 224 13 of 27 Working and manipulating brain cancer images to recognize the hidden features inside the
images and subsequently detect/classify/predict relevant diseases are extremely essential for clinicians,
and noninvasive methods including CNN-based classifiers can do this task very well. In general,
CNN-based classifiers might be applied to three different kinds of brain cancer images including H&E
histology, MR, or both. These classifiers can categorize various brain tumor types or predict biomarker
status and patients’ survival rate based on the features inside the images. Hence, two separated
and important parts can be considered to achieve the best results: preprocess and CNN architecture. Based on the various studies that we completed, some steps which play important roles in the
preprocessing part include patch extraction, abnormal patch detection/selection, intensity normalization,
contrast enhancement, and background removing. Overall, there is no best practice to recommend
one or several steps in the preprocessing phase and researchers have to try many experiments to find
the best results. However, studies show that data augmentation techniques to expand the number
of data samples as model inputs have successfully helped the CNNs to achieve higher accuracy. In the second part, CNN models can be first trained to extract features from brain cancer images,
and then, those feature vectors along with patients’ clinical data together make a numerical dataset. Afterwards, some linear regression models or nonlinear machine learning methods use this dataset to
classify/predict relevant biomarker status, survival rate, or brain tumor types. However, It would be
simpler to directly apply CNNs as a classifier or predictive model (Figure 1). 3. DCNNs Application in the Segmentation of Brain Cancer Images In this section, the application of DCNNs to extract the most important features from different
kinds of brain cancer images for detection/segmentation task is explained. For this purpose and more
simplicity, we have divided this part into three subsections where in each one a specific image type i.e.,
H&E histology, MRI, or CT, is explored. 3.1. DCNNs Application in the Segmentation of Brain Cancer MR Images Ismael et al. [69] applied the DCNN ResNet50 architecture to segment glioma, meningioma,
and pituitary tumor tissues of 3064 MRI images from 233 patients. ResNet50 uses skip connections to
avoid gradient degradation, and thereby enables training of much deeper networks than previously
thought possible. Due to limited sample in the training dataset, data augmentation was used to
generate additional training data to improve results. As the distribution of segmented classes is not
uniform, accuracy alone is not a suitable measure of performance. Instead, a combination of accuracy,
precision, recall, F1-score, and the balanced accuracy were used, which were 97%, 98%, 97%, 97%,
and 97%, respectively, at a patient-level. Maharjan et al. [70] propose an enhanced softmax loss function. The model was applied to
glioma, meningioma, and pituitary tumor segmentation of 3064 MRI images. The resulting loss
function utilizes regularization and is far more suitable for multiclass classification than traditional
loss functions. This difference helps avoid overfitting when compared to the traditional sigmoid loss
function. Using the enhanced softmax loss function, the accuracy was improved to 99.54% 98.14%,
and 98.67% for meningioma, glioma, and pituitary tissues, respectively. Regularization also improved
runtime by 40–50 ms per sample. A novel brain tumor segmentation architecture was proposed by Vijh et al. [71] that employs
a blend of Otsu thresholding, Adaptive Particle Swarm Optimization (APSO), and morphological
operations in the skull stripping preprocessing steps. Skull stripping removes noncerebral tissue not
needed for analysis and is a crucial step in neurological imaging. Once preprocessed, 19 features
(cluster prominence, cluster shade, contrast, etc.)
are extracted from the cerebral tissue image
using GLCM (Grey Level Co-occurrence Matrix). These features are passed into a densely connected
three-layer CNN. The model yields 98% accuracy applied to the Internet Brain Segmentation Repository
(IBSR) dataset, which consists of 18 samples of various tumor types. J. Pers. Med. 2020, 10, 224 14 of 27 Rani et al. [72] propose a method that utilizes adaptive thresholding and high boost convolution
filtering to segment brain tumors. Preprocessing steps first extract 2D slices from the 3D MRI scans
and any grainy noise is filtered using averaging techniques. The preprocessed dataset is input into the
segmentation state which applies Region Growing & Local Binary Pattern (LBP) operators to build
a feature vector. 3.1. DCNNs Application in the Segmentation of Brain Cancer MR Images This is then fed into the mask generation stage that applies the Fuzzy C-Means
Algorithm, Otsu Thresholding, and high boost convolution filtering to extract the high energy glioma
regions. The resulting model scored 87.20% for HGG on 210 samples and 83.77 for LGG on 75 samples. g
g
p
p
Deng et al. [73] propose a novel architecture to segment FLAIR, Tc1, and T2 images from the
BraTS2013 and 2015 datasets of over 270 HGG and LGG scans. The architecture composes HCNN and
CRF-RRNN models to segment. The HCNN creates image slices at mixed scales to better leverage
location and context at a greater scale. The HCNN model also fuses axial, coronary, and sagittal images. The CRF-RRNN takes the output of the HCNN and produces a global segmentation based on the slices
input into the HCNN. The resulting model scored 98.6% accuracy. Deng et al. [74] improve upon the FCNN network. Batch normalization is added to the network to
improve computational performance. The addition of Dense Micro-block Difference features also assist
with spatial consistency. Utilizing Fisher vector encoding methods better invariance to texture rotation
and scale. The model achieved 91.29% average Dice score on the BraTS2015 dataset on 220 HGG and
50 LGG scans. Kumar et al. [75] propose a 3D CNN architecture that focuses on correcting intensity inhomogeneity. This is achieved through a preprocessing step that utilizes a novel N3T-spline to correct bias field
distortion for reducing noise and intensity variation in the 3D scans. The N3T-spline utilizes the
standard N3 (nonuniformity non-parametric normalization) framework but uses a T-spline smoothing
strategy. A grey level co-occurrence matrix (GLCM) layer extracts feature vectors from the preprocessed
scans. The feature vectors are input into the novel 3D CNN and a simple thresholding scheme is
applied to correct false labels and any other undesired noise. The resulting model scored competitively
on the BraTS2015 dataset. The dataset consists of 220 HGG scans and 50 LGG scans. Mittal et al. [76] propose an architecture utilizing Stationary Wavelet Transform (SWT) and Growing
Convolution Neural Network (GCNN) to segment neurological images. The model preprocesses input
with Wiener filtering and Otsu Thresholding to remove noise and convert to a binary image. A novel
skull stripping algorithm is proposed that leverages blob detection and labelling methods to more
effectively remove skill, fat, and skin from the regions of interest. 3.1. DCNNs Application in the Segmentation of Brain Cancer MR Images Once the images are preprocessed,
features are extracted with SWT and classified with a Random Forest implementation. The GCNN then
encodes the classified features into a segmented output. The model was able to achieve an Structural
similarity (SSIM) score of 98.6% on the BRAINIX dataset of 2457 scans. Mittal et al. [77] composed a dataset of MRI scans of Glioma, Meningioma, Pituitary, and Negative
brain tumor results. The dataset was used to evaluate the performance of several common pre-trained
CNN architectures (simple CCNs, VGG, and Xception). Next, several of the pretrained models
were fused together in a composite architecture, specifically using simple CNNs, Xception, VGG16,
and VGG19. To avoid overfitting, augmented data was utilized in some branches of the architecture,
scaling the dataset from 1167 to 5835 samples. The composite architecture achieved an accuracy
of 98.89%. Thillaikkarasi et al. [78] proposed a deep learning model to classify and segment MRI scans. Images are first preprocessed with LoG and Contrast Adaptive Histogram Equalization (CLAHE)
filters. Preprocessed scans are fed into a Spatial Gray Level Dependency Matrix (SGLDM) and
the following features are generated: contrast, mean, variance, entropy, energy and homogeneity. The feature set is input into a multiclass-SVM (M-SVM) and the MRI scan is classified as abnormal or
normal. Abnormal images are then input into a CNN to segment brain tumors from healthy tissues. The resulting model scored 84% accuracy on 40 MRI scans. g
y
Sharma et al. [79] proposed a novel skull stripping algorithm that utilizes Differential Evolution
(DE) and Otsu Thresholding. The algorithm first normalizes the input images and applies a Gaussian J. Pers. Med. 2020, 10, 224 15 of 27 filter. Next, a global threshold is calculated and iteratively optimized using Otsu Thresholding and DE
method. Morphological operations are then applied to extract the neurological tissues. Features are
extracted from the preprocessed images with GLCM. The features used in this model are contrast,
energy, entropy, correlation, standard deviation. The feature set is used to train the CNN with the
Network Fitting Tool (nf-tool). The model is trained on the IBSR dataset, containing 18 samples of
various tumors, and the MS-Free dataset, containing 38 tumor free samples. The model is able to
achieve 94.73% accuracy. y
Kong et al. [80] expand on the traditional UNet architecture and propose the Hybrid Pyramid
U-Net model (HPU-Net). 3.1. DCNNs Application in the Segmentation of Brain Cancer MR Images Key additions are batch normalization to the downsampling component
of the network to help combat vanishing gradient during the training process. This is particularly
important for brain tumor segmentation to avoid missing any small lesions that would otherwise
be missed. In the upsampling component, bilinear interpolation is used in favour of convolution
transposing of convolutional layers as to not add additional parameters. Additionally, semantic and
location features are emitted in each upsampling block and combined to capture multiscale information. This effectively creates a feature pyramid to capture brain tumor lesions with multiscale shapes and
sizes. The resulting network was trained on BraTS2015 and BraTS2017 on a collective 430 HGG and
145 LGG images. The network achieved 71% and 80% Dice score, respectively. Benson et al. [81] apply a modified Hourglass Network to brain tumor segmentation. The original Hourglass Network is an encoder–decoder architecture with several residual blocks. Through experimentation, it was determined that using five downsampling layers instead of seven
performed better and was computationally less intensive. A single residual block per level was used
instead of two. It was also determined that concatenating layers followed by a 1x1 convolution layer
outperformed the original element-wise summation, despite additional memory usage. The resulting
architecture yielded 92% accuracy on BraTS2018 of 210 HGG and 75 LGG scans. Zhou et al. [82] propose an ensemble network with several variations on Model Cascade (MC) Net
and One-Pass Multi-Task (OM) Net. Modifications made to the MC-Net include an additional series of
nested and dense skip layers to improve the feature map coverage of the encoder-decoder architecture. Another MC-Net variation includes adding multiscale contextual information by including inputs
at different scales to more effectively extract semantic features at different resolutions. Variations of
the OM-Net include making a deeper model by appending an additional residual block to the
original OM-Net. Variations to both networks include adding “Squeeze-and-Excitation” (SE) attention
mechanisms to adaptively model interdependencies between channel-wise features. This effectively
increases sensitivity to informative features and decreases sensitivity to others. Each model and
variation were separately trained and then ensembled into a single model. The resulting model
achieved 90% accuracy on whole tumor segmentation on the BraTS2018 dataset, consisting of 210 HGG
and 75 LGG scans. Dai et al. [83] implement an ensemble model consisting of a modified U-Net model and a Domain
Adaptive U-Net (DAU-Net) model. 3.1. DCNNs Application in the Segmentation of Brain Cancer MR Images The modified U-Net uses five residual blocks in both the encoding
and decoding components of the network. Group normalization is also applied to add stability
given small batches. The DAU-Net is structurally the same as the modified U-Net except instance
normalization is applied instead since it is experimentally shown to boost domain adaptation. In total,
nine variations of the mentioned models were trained with varying preprocessing techniques and
fused together using XGBoost. The model was trained on BraTS2018 consisting of 210 HGG and
75 LGG scans. The resulting ensemble network scored 91% accuracy on whole tumor segmentation. Kermi et al. [84] proposed a network based on the U-Net architecture. The modified architecture
introduces three residual blocks in both the encoding and decoding phases of the architecture. Unlike the
original U-Net blocks, each encoding block uses batch normalization and a Parametric Rectified Linear
Unit (PReLU). A convolution layer with a stride of two is applied for downsampling. The novel
U-Net architecture achieved an 86.8% Dice score on the BraTS2018 dataset consisting of 210 HGG and
75 LGG images. Dai et al. [83] implement an ensemble model consisting of a modified U-Net model and a Domain
Adaptive U-Net (DAU-Net) model. The modified U-Net uses five residual blocks in both the encoding
and decoding components of the network. Group normalization is also applied to add stability
given small batches. The DAU-Net is structurally the same as the modified U-Net except instance
normalization is applied instead since it is experimentally shown to boost domain adaptation. In total,
nine variations of the mentioned models were trained with varying preprocessing techniques and
fused together using XGBoost. The model was trained on BraTS2018 consisting of 210 HGG and
75 LGG scans. The resulting ensemble network scored 91% accuracy on whole tumor segmentation. Kermi et al. [84] proposed a network based on the U-Net architecture. The modified architecture
introduces three residual blocks in both the encoding and decoding phases of the architecture. Unlike the
original U-Net blocks, each encoding block uses batch normalization and a Parametric Rectified Linear
Unit (PReLU). A convolution layer with a stride of two is applied for downsampling. The novel
U-Net architecture achieved an 86.8% Dice score on the BraTS2018 dataset consisting of 210 HGG and
75 LGG images. J. Pers. Med. 2020, 10, 224 16 of 27 Mlynarski et al. [85] extend the classic U-Net model to train with “mixed supervision”. 3.1. DCNNs Application in the Segmentation of Brain Cancer MR Images Mixed supervision is defined as a dataset with some images fully-annotated (pixel-wise ground
truth segmentation) and weakly-annotated (image-level label denoting the presence of a tumor). The model was trained with 210 HGG and 75 LGG fully annotated MRI scans from the BraTS2018
dataset. As a result, the extended U-Net now has an additional subnetwork that performs image-level
classification that determines if the scan has a tumor or not. The additional subnet allows the network
to exploit additional samples that are only weakly-annotated. The extended model is also expanded
to the multiclass problem to segment several classes of interest: nontumor, contrast-enhancing core,
edema, nonenhancing core. The proposed model was shown to output a more accurate segmented
image when provided weakly annotated samples. Wang et al. [86] introduce a unique architecture Brain Image-specific Fine-tuning Segmentation
(BIFSeg) to address zero-shot learning in medical image segmentation. Zero-shot learning is the
machine learning problem when the model encounters classes not observed during training. BIFSeg,
based on the P-Net architecture, takes a bounding box as user input localized to the desired tumor core. Once segmented, further optional user input as scribbles can be used to fine-tune the segmentation. When applied to BraTS2015, the model achieved 86.29% Dice score on 220 HGG and 50 LGG scans. 3.2. DCNNs Application in the Segmentation of Brain Cancer CT Images Monteiro et al. [87] applied a 3D CNN architecture to Traumatic Brain Injury (TBI) CT scans to
achieve voxel-wise segmentation into multiclass lesions. The methodology was to first train the CNN
on CENTER-TB1 Dataset 1 for initial results & weightings. Then, the CENTER-TB1 Dataset 2 was
incorporated and some segmentations were manually corrected. The resulting model was then applied
to the CQ500 dataset and achieved a 94% AUC accuracy on over 1000 combined CT scans. Though the
overall accuracy of this model is slightly lower than other state-of-the-art models, the proposed model
has the added ability to distinguish between different lesion types and progression to better understand
and personalize care, which is very important in traumatic brain injuries. 3.3. DCNNs Application in the Segmentation of Brain Cancer H&E Histology Images A standard histopathology scan is on the range of 100,000 × 100,000 pixels. This large scale
makes training such models very difficult. Xu et al. [88] proposed a unique approach to supporting
classification and segmentation on said large-scale brain tumor histopathology. Their architecture
extracts 112 × 112 patches from the original image. The set of patches becomes the input to the
ImageNet LSVRC 2013 architecture, generating a 4096-dimensional feature vector per extracted patch. Feature vectors are pooled into a linear SVM classifier to produce probability maps to distinguish
necrosis from healthy tissue. The resulting architecture yielded 84% accuracy on the MICCAI 2014
dataset, consisting of 35 samples. All information related to the segmentation task mentioned above is summarized in Table 2. We have also depicted in Figure 2 the main steps of this part as a roadmap for researchers who are
going to apply DCNNs in their future work for the segmentation task. 17 of 27 J. Pers. Med. 2020, 10, 224 Table 2. DCNN-based segmentation models brief description (sorted by year published//DCNN performance). Ref
Year
Tumor Type
Task
Model Name
Image Type
Model Desc. Software
Hardware
Dataset
Instances
Performance
[70] **
2020
Glioma,
Meningioma,
Pituitary
Segmentation
ELM-LRF
MRI
Implemented an enhanced softmax
loss function that is more suitable
for multiclass applications. Python 3.6; Keras
–
2.8 GHz Intel Core i7
7th gen processor with
16 GB RAM and 4 GB
–
NVIDIA 1050 memory
Brain Tumor
Dataset [58]
3064
99.54%,
98.14%,
98.67%
(Per Tumor
Type)
[69]
2020
Glioma,
Meningioma,
Pituitary
Segmentation
ResNet50
MRI
Glioma, meningioma, and pituitary
tumor segmentation with the
ResNet50 architecture. –
Python 3.6;
Keras 2.2.4;
–
Tensorflow
1.13
–
NVIDIA GeForce
RTX 2070
–
GPU; Intel i5-9600K @
3.7 GHz and 16
GB RAM
Brain Tumor
Dataset [58]
3064
99%
[73]
2020
Glioma
Segmentation
HCNN; CRF-RRNN
MRI
The composite architecture of
HCNN to capture mixed scale
context and CRF-RRNN
reconstruct a global segmentation. N/A
N/A
–
BraTS2013
[59]
–
BraTS2015
[64]
220 HGG;
50 LGG
98.6%
[74]
2019
Glioma
Segmentation
FCNN; DMD
MRI
Enhanced FCNN with batch
normalization and DMD features to
provide spatial consistency. Fisher
vector encoding method for texture
invariance to scale and rotation. 3.3. DCNNs Application in the Segmentation of Brain Cancer H&E Histology Images Caffe
–
CPU Intel Core i7
–
3.5GHz, GPU NVIDIA
GeForce GTX1070
BraTS2015 [64]
220 HGG;
50 LGG
91%
[86] **
2018
Glioma
Segmentation
P-Net; PC-Net
MRI
Addresses zero-shot learning by
taking user input bounding boxes
and scribbles to
fine-tune segmentations. Caffe
–
2 8-core
E5-2623v3 Intel
–
Haswell, a K80
NVIDIA GPU and
128GB memory
BraTS2015 [64]
220 HGG;
50 LGG
86.29%
[75]
2019
Glioma
Segmentation
FCNN
MRI
A novel N3T-spline utilizes is used
to preprocess 3D input images. GLCM extracts feature vectors and
are inputs into a CNN. MATLAB R2017a
N/A
BraTS2015 [64]
220 HGG;
50 LGG
N/A
[71]
2020
N/A
Segmentation
3-layer DCNN *
MRI
Utilized Otsu thresholding to create
a novel skull stripping algorithm. GLCM and a three-layer CNN
segments the stripped images. MATLAB R2018b
N/A
IBSR [89]
18
98% Table 2. DCNN-based segmentation models brief description (sorted by year published//D . DCNN-based segmentation models brief description (sorted by year published//DCNN performance). J. Pers. Med. 2020, 10, 224 18 of 27 Table 2. Cont. Ref
Year
Tumor Type
Task
Model Name
Image Type
Model Desc. Software
Hardware
Dataset
Instances
Performance
[72]
2020
Glioma
Segmentation
Automatic Detection
and Segmentation of
Tumor (ADST) *
3D MRI
Region Growing and Local Binary
Pattern (LBP) operators are used to
build a feature vector that is
then segmented
N/A
N/A
BraTS2018 [61]
210 HGG;
75 LGG
87.20% for
HGG; 83.77
for LGG
(Average
Jaccard)
[81] **
2019
Glioma
Segmentation
Hourglass Net
MRI
Enhanced Hourglass Network with
added residual blocks and novel
concatenation layers. N/A
NVIDIA TITAN X GPU
BraTS2018 [61]
210 HGG;
75 LGG
92%
[83] **
2019
Glioma
Segmentation
XGBoost; U-Net;
DAU-Net (Domain
Adaptive U-Net) *
MRI
Implementation of a U-Net
variation using instance
normalization to boost
domain adaptation. PyTorch
4 NVIDIA Titan Xp GPU
cards
BraTS2018 [61]
210 HGG;
75 LGG
91% (Whole
Tumor)
[82]
2019
Glioma
Segmentation
MC-Net; OM-Net
MRI
Ensemble network of several
MC-Net and OM-Net variations. Attention mechanisms are added to
increases sensitivity to relevant
channel-wise interdependencies
N/A
N/A
BraTS2018 [61]
210 HGG;
75 LGG
90% (Whole
Tumor)
[84] **
2019
Glioma
Segmentation
U-Net
MRI
The U-Net variation that uses batch
normalization and residual blocks
to improve performance on
neurological images. 4. Discussion As seen in the many articles that we have reviewed, DCNNs with classification and segmentation
architectures have proven to be very powerful and effective when applied to medical imaging in
retrospective studies. Specifically, it has been demonstrated that using pretrained models and transfer
learning from generic applications to medical imaging applications saves time and achieves better
model performance. In particular, when applied to the analysis of histopathological and MR images
for brain cancer and is useful as a predictive tool for analysis of patients’ survival and their correlation
with cancer genetic signatures. The reviewed literature provides a roadmap for future classification and
segmentation methods and provides examples of their different type of applications. Although there
is no best practice or specific guidelines to design and implement DCNN-based classification or
segmentation models, some studies reviewed have proved that by applying preprocessing stages such
as data augmentation, background removing, noise reduction, intensity normalization, and patch
extraction, as well as choosing the appropriate architectures as classification and segmentation models,
the performance of DCNN models might be dramatically increased. Figures 1 and 2 summarize the
most important points recommended by reviewed works to achieve the best results with DCNNs. 3.3. DCNNs Application in the Segmentation of Brain Cancer H&E Histology Images Shows suggested roadmap and stages of a typical brain tumor segmentation Architecture
with the following high-level steps: 1. Input: magnetic resonance (MRI) and computed tomography
(CT) scans are input into the model; 2. Preprocessing: apply several techniques to normalize images,
remove noise, and filter irrelevant components; Step 3: Deep Convolutional Neural Network (DCNN)
Application: The preprocessed dataset is fed into a DCNN model the extract features for segmentation,
with localization a key component; 4. Output Images: Specifies the result of the segmentation model. Figure 2. Shows suggested roadmap and stages of a typical brain tumor segmentation Architecture
with the following high-level steps: 1. Input: magnetic resonance (MRI) and computed tomography
(CT) scans are input into the model; 2. Preprocessing: apply several techniques to normalize images,
remove noise, and filter irrelevant components; Step 3: Deep Convolutional Neural Network (DCNN)
Application: The preprocessed dataset is fed into a DCNN model the extract features for segmentation,
with localization a key component; 4. Output Images: Specifies the result of the segmentation model. As explored in the several studies, methods for detecting and segmenting brain tumor tissues can
take several forms. In general, architectures follow the following three stages: preprocess, segmentation,
and postprocess. In the preprocessing stages, several techniques have proven successful including Otsu
thresholding, patch extraction, noise reduction, morphological operations, and intensity normalization. Outside of these standard processes, several novel extensions of these standard practices have been
developed including variations on Otsu thresholding. Furthermore, during the training segmentation
models, data augmentation techniques have been successful to expand limit datasets to make deep
learning feasible. Several models and variations on classic models including Xception Net, U-Net,
and SVM with very promising outputs result in brain tumor detection and segmentation (Figure 2). 3.3. DCNNs Application in the Segmentation of Brain Cancer H&E Histology Images N/A
–
Intel Xeon E5-2650
CPU@ 2.00 GHz (64
GB) and
NVIDIA Quadro
–
4000–448 Core CUDA
(2 GB) GPU
BraTS2018 [61]
210 HGG;
75 LGG
86.8%
(Whole
Tumor)
[85]
2019
Glioma
Segmentation
U-Net
MRI
Extension of the U-Net to train with
“mixed supervision”, meaning both
pixel-wise & image-level ground
truths to achieve
superior performance. N/A
N/A
BraTS2018 [61]
210 HGG;
75 LGG
N/A for the
entire dataset J. Pers. Med. 2020, 10, 224 19 of 27 Table 2. Cont. Table 2. Cont. Ref
Year
Tumor Type
Task
Model Name
Image Type
Model Desc. Software
Hardware
Dataset
Instances
Performance
[77]
2019
Glioma,
Meningioma,
Pituitary, and
Negative
Classification
–
CNN,
–
Xception,
VGG16, VGG19
MRI
Several of the pre-trained models
(simple CNNs, Xception, VGG16,
and VGG19) were fused together in
a composite architecture. Keras
N/A
N/A
1167
98.89%
[87] **
2020
TBI (Traumatic
Brain Injury)
Segmentation
CNN
CT
3D CNN architecture to create
voxel-wise segmentation of TBI
CT scans. N/A
N/A
CENTER-TBI
(Datasets 1 & 2)
[90]; CQ500 [91]
539; 500
(Patients)
94%
[78]
2019
N/A
Segmentation
M-SVM;
CNN
MRI
SGLDM and M-SVM are applied to
extract and classify MRI scans. CNN is then applied to segment
the extracted feature vectors. N/A
N/A
N/A
40
84%
[76]
2019
N/A
Segmentation
SWT; GCNN
MRI
Dataset is preprocessed with a
novel skull stripping. Features are
extracted with SWT, classified with
a Random Forest implementation
and finally segmented with GCNN. N/A
N/A
BRAINIX [92]
2457
98.6% (SSIM
Score)
[80] **
2018
Glioma
Segmentation
U-Net
MRI
HPU-Net enhances the traditional
U-Net with multiscale images and
image pyramids. Keras; Tensorflow
NVIDIA Titan X GPU
–
BraTS2015
[64]
–
BraTS2017
[93]
430 HGG;
145 LGG
71% and 80%
(Respective
to dataset)
[88] **
2015
Glioma
Segmentation
ImageNet LSVRC 2013
H&E
Histology
Patches are extracted from large
histopathology scans and passed
into ImageNet LSVRC 2013
architecture. Linear SVM classifier
pools extracted feature vectors. N/A
N/A
MICCAI 2014
[94]
35
84%
* Model names with asterisks are not defined in the original papers and names were assigned based on the models applied. Note: For abbreviations description in this table please refer to
the list of abbreviations on the back part of this article (before References). ** the references with “**” mean that the results achieved by their methods or the dataset used have been
validated/supervised by specialists (e.g., pathologists/radiologists). J. Pers. Med. 3.3. DCNNs Application in the Segmentation of Brain Cancer H&E Histology Images 2020, 10, 224
expand limit datase
including Xception 20 of 27
ic models
detection Figure 2. Shows suggested roadmap and stages of a typical brain tumor segmentation Architecture
with the following high-level steps: 1. Input: magnetic resonance (MRI) and computed tomography
(CT) scans are input into the model; 2. Preprocessing: apply several techniques to normalize images,
remove noise, and filter irrelevant components; Step 3: Deep Convolutional Neural Network (DCNN)
Application: The preprocessed dataset is fed into a DCNN model the extract features for segmentation,
with localization a key component; 4. Output Images: Specifies the result of the segmentation model. Figure 2. Shows suggested roadmap and stages of a typical brain tumor segmentation Architecture
with the following high-level steps: 1. Input: magnetic resonance (MRI) and computed tomography
(CT) scans are input into the model; 2. Preprocessing: apply several techniques to normalize images,
remove noise, and filter irrelevant components; Step 3: Deep Convolutional Neural Network (DCNN)
Application: The preprocessed dataset is fed into a DCNN model the extract features for segmentation,
with localization a key component; 4. Output Images: Specifies the result of the segmentation model. Figure 2. Shows suggested roadmap and stages of a typical brain tumor segmentation Architecture
with the following high-level steps: 1 Input: magnetic resonance (MRI) and computed tomography
Figure 2. Shows suggested roadmap and stages of a typical brain tumor segmentation Architecture Figure 2. Shows suggested roadmap and stages of a typical brain tumor segmentation Architecture
with the following high-level steps: 1. Input: magnetic resonance (MRI) and computed tomography
(CT) scans are input into the model; 2. Preprocessing: apply several techniques to normalize images,
remove noise, and filter irrelevant components; Step 3: Deep Convolutional Neural Network (DCNN)
Application: The preprocessed dataset is fed into a DCNN model the extract features for segmentation,
with localization a key component; 4. Output Images: Specifies the result of the segmentation model. Figure 2. Shows suggested roadmap and stages of a typical brain tumor segmentation Architecture
with the following high-level steps: 1. Input: magnetic resonance (MRI) and computed tomography
(CT) scans are input into the model; 2. Preprocessing: apply several techniques to normalize images,
remove noise, and filter irrelevant components; Step 3: Deep Convolutional Neural Network (DCNN)
Application: The preprocessed dataset is fed into a DCNN model the extract features for segmentation,
with localization a key component; 4. Output Images: Specifies the result of the segmentation model. Figure 2. 5. Challenges and Future Considerations Deep learning techniques in the medical imaging space come with several challenges. Firstly,
there is currently no agreed-upon framework or methodology for selecting model hyperparameters,
such as weight decay, residual blocks, dropout, number of hidden layers, and image input size. Instead, hyperparameters need to be determined experimentally. Secondly, within current medical 21 of 27 J. Pers. Med. 2020, 10, 224 imaging, dataset curation requires expensive and time-consuming work to be done by specialists e.g.,
radiologists and pathologists which may not be possible in every circumstance. The resulting datasets
are poor quality and of an insufficient quantity, requiring heavy augmentation to achieve the high
number of samples for DCNNs. Moreover, most of the studies are undertaken using retrospective
datasets which leave uncertainty about the extent to which these models can be applied to newly
generated data and whether this newly generated data can be used for training and testing of these
models. Furthermore, it is becoming widely accepted, that at the very early stages of this type of project
(i.e., during the model conception), an often strong, interdisciplinary collaboration between medical
imaging professionals and machine learning experts is required. Image preprocessing techniques,
such as skull stripping and thresholding, are widely used but no standard algorithms or steps have
been defined, even for common datasets or types of medical image. This limits the extent to which
these procedures can be applied by nondeep learning experts. Finally, DCNN models applied to
segmentation and classification analysis of biomedical imaging have not been extensively integrated
with all the available clinical metadata. For example, TCGA data sets contain patient demographics
data, tumor RNAseq data, survival data, as well as pathology and RNAseq images, but relatively little
has been explored in these areas, except for the very few cases detailed in our review. p
p
y
When considering future directions, despite the advancements of DCNNs, we still face challenges
in translating into clinical practice the use of DCNN-based classification and segmentation methods. So far, many studies have shown the powerful capacity of DCNN methods but still, we lack prospective
studies that can further validate them. We envisage a future where these methods are incorporated into
clinical trials. These could enable the running and testing of different DCNN models (already created)
and measure their relative efficacy on tumor classification and segmentation. 5. Challenges and Future Considerations In this regard, we believe
this survey could be an important source of DCNN application to brain cancer images. It could provide
a toolbox for the inclusion of such methods in these studies. As we have seen, classification methods
have been only been applied to a few groups of brain cancer patients (e.g., IDH and mutant). However,
cancer heterogeneity is much more complex, with four subtypes (Proneural, classical, mesenchymal,
and neural, although the neural subtype may be nontumor specific) [95]. Therefore, any uncertainty
regarding the applicability of DCNN models on nonstratified analysis can be overcome by better
designed clinical studies that implement these methodologies. Despite the accuracy of segmentation
approaches increasing significantly, this has not been translated to molecular information regarding
transcriptomic profiles and genetic signatures associated with different tumor regions. If available,
this would permit the determination of the cellular composition of the tumor microenvironment and
tumor-stroma interactions that are important in driving cancer progression. Thus, there are still several
areas that can be improved to be able to translate deep learning methodologies into clinical medicine. We believe that future studies which address these issues and formulate standard pipelines to deliver
performance and accuracy on reliable datasets will be important to define reference standards for
testing and validating new DCNN approaches. 6. Conclusions The ability for physicians to quickly and accurately classify and segment brain tumor scans has
never been more important. Breakthroughs in deep learning and DCNN architectures applied to
biomedical radiology have almost brought these functions into practice. In this study, we have reviewed the most novel research articles on deep learning applied to
medical imaging problems, specifically brain tumor classification and segmentation tasks. The review
has resulted in the creation of a roadmap (summarized in Figures 1 and 2 above) for resolving both
tasks. This can be leveraged by researchers for implementing models of their own. In addition,
Tables 1 and 2 present a compilation of relevant information, applied techniques, deep learning
networks, and DCNN-based models’ performance for the future development of research in this area. J. Pers. Med. 2020, 10, 224 22 of 27 Author Contributions: Data collection, writing—original draft preparation, review and editing, pictures and
tables illustration, A.Z.S., E.F., and G.A.G.; funding acquisition, G.A.G.; supervision and project management,
G.A.G., M.D.M., and M.Y.; revision and quality improvement, M.D.M., Y.C., L.T.-O., and D.I. All authors have
read and agreed to the published version of the manuscript. Funding: This work was supported by grants from the National Health and Medical Research Council of Australia
(1067405 and 1123816 to G.A.G.), the Cure Brain Cancer Foundation (to G.A.G.), the University of South Australia
(to G.A.G. and A.Z.S.), the Neurosurgical Research Foundation (to G.A.G.) and the Cancer Council SA Beat Cancer
Project Infrastructure (to G.A.G.). G.A.G. is also supported by an Australian Research Council Future Fellowship
(FT160100366). A.Z.S. is supported by an Australian Government Research Training Program (RTP) Scholarship. Conflicts of Interest: The authors declare no conflict of interest. References 1. Shirazi, A.Z.; Mohammadi, Z. A hybrid intelligent model combining ANN and imperialist competitive
algorithm for prediction of corrosion rate in 3C steel under seawater environment. Neural Comput. Appl. 2017, 28, 3455–3464. 1. Shirazi, A.Z.; Mohammadi, Z. A hybrid intelligent model combining ANN and imperialist competitive
algorithm for prediction of corrosion rate in 3C steel under seawater environment. Neural Comput. Appl. 2017, 28, 3455–3464. 2. Shirazi, A.Z.; Hatami, M.; Yaghoobi, M.; Chabok, S.J.S.M. An intelligent approach to predict vibration rate in
a real gas turbine. Intell. Ind. Syst. 2016, 2, 253–267. 2. Shirazi, A.Z.; Hatami, M.; Yaghoobi, M.; Chabok, S.J.S.M. An intelligent approach to predict vibration rate in
a real gas turbine. Intell. Ind. Syst. 2016, 2, 253–267. 3. Shirazi, A.Z.; Tofighi, M.; Ganjefar, S.; Mahdavi, S.J.S. An optimized adaptive-neuro fuzzy inference system
(ANFIS) for reliable prediction of entrance length in pipes. Int. J. Enhanc. Res. Sci. Technol. Eng. 2014, 3, 79–89. 3. Shirazi, A.Z.; Tofighi, M.; Ganjefar, S.; Mahdavi, S.J.S. An optimized adaptive-neuro fuzzy inference system
(ANFIS) for reliable prediction of entrance length in pipes. Int. J. Enhanc. Res. Sci. Technol. Eng. 2014, 3, 79–89. 4. Yavuz, E.; Eyupoglu, C. An effective approach for breast cancer diagnosis based on routine blood analysis
features. Med. Biol. Eng. Comput. 2020, 58, 1583–1601. 4. Yavuz, E.; Eyupoglu, C. An effective approach for breast cancer diagnosis based on routine blood analysis
features. Med. Biol. Eng. Comput. 2020, 58, 1583–1601. 5. Becker, A. Artificial intelligence in medicine: What is it doing for us today? Health Policy Technol. 2019, 8, 198–205. . Shirazi, A.Z.; Fornaciari, E.; Gomez, G.A. Deep learning in precision medicine. In Artificial Intelligen
Precision Health; Elsevier: Amsterdam, The Netherlands, 2020; pp. 61–90. 7. Shirazi, A.Z.; Chabok, S.J.S.M.; Mohammadi, Z. A novel and reliable computational intelligence system for
breast cancer detection. Med. Biol. Eng. Comput. 2018, 56, 721–732. 8. Zhu, W.; Xie, L.; Han, J.; Guo, X. The Application of Deep Learning in Cancer Prognosis Prediction. Cancers
2020, 12, 603. 9. Coccia, M. Deep learning technology for improving cancer care in society: New directions in cancer imaging
driven by artificial intelligence. Technol. Soc. 2020, 60, 101198. 10. Huang, Z.; Johnson, T.S.; Han, Z.; Helm, B.; Cao, S.; Zhang, C.; Salama, P.; Rizkalla, M.; Yu, C.Y.; Cheng, J.;
et al. Deep learning-based cancer survival prognosis from RNA-seq data: Approaches and evaluations. BMC Med. Genom. 2020, 13, 1–12. 11. Baptista, D.; Ferreira, P.G.; Rocha, M. Abbreviations Abbreviations
AI
Artificial Intelligence
ANN
Artificial Neural Network
APSO
Adaptive Particle Swarm Optimization
AUC
Area Under the Curve
BraTS-xxxx
Brain Tumour Segmentation Challenge-year
CapsNets
Capsule Networks
CLAHE
Contrast Adaptive Histogram Equalization
CRF
Conditional Random Forest
CT
Computed Tomography
DCNN
Deep Convolutional Neural Network
DenseNet
Densely Connected Convolutional Networks
DMD
Dense Micro-block Difference
DNN
Deep (Artificial) Neural Network
DWT
Discrete Wavelet Transform
ELM
Extreme Learning Machines
FA
False Alarm
FC
Fully Connected
FLAIR
T2-Weighted Fluid-Attenuated Inversion Recovery
GAN
Generative Adversarial Networks
GBM
Glioblastoma
GCNN
Growing Convolution Neural Network
GD
Gradient Descent
GLCM
Grey Level Co-occurrence Matrix
H&E
Haematoxylin and Eosin
HCNN
Heterogenous Convolution Neural Network
HGG
High-Grade Glioma
IDH
Isocitrate Dehydrogenase
LBP
Local Binary Pattern
LGG
Low-Grade Glioma
LoG
Laplacian of Gaussian
LRF
Local Receptive Fields
LSTM
Long Short-Term Memory
MA
Missed Alarm
MC-Net
Model Cascade Net
ML
Machine Learning
MR
Magnetic Resonance
M-SVM
Multiclass-Support Vector Machine
N/A
Not Available
OM-Net
One-Pass Multi-Task Net
PCA
Principal Component Analysis
Pr
Predictive Model
ResNet
Residual Network
RNN
Recurrent Neural Network 23 of 27 J. Pers. Med. 2020, 10, 224 RoI
Region of Interest
RRNN
Recurrent Regression-based Neural Networks
SVC
Support Vector Classifier
SWT
Stationary Wavelet Transform
T1 (T1-Weighted)
Longitudinal Relaxation Time
T1c
T1-Weighted Post-Contrast
T1-Gado
Gadolinium contrast medium in MR images
T2 (T2-Weighted)
Transverse Relaxation Time
TBI
Traumatic Brain Injury
TCGA
The Cancer Genome Atlas
TCIA
The Cancer Imaging Archive
WBA
The Whole-Brain Atlas
WSI
Whole Slide Image
xD
x Dimensional References 2020, 40, 1225–1232. [CrossRef]
22. Mzoughi, H.; Njeh, I.; Wali, A.; Slima, M.B.; BenHamida, A.; Mhiri, C.; Mahfoudhe, K.B. Deep Multi-Scale
3D C
l
i
l N
l N
k (CNN) f
MRI Gli
B
i
T
Cl
ifi
i
J Di i
I
i g
on deep learning using hybrid model CNN and NADE. Biocybern. Biomed. Eng. 2020, 40, 1225–1232. [CrossRef]
22. Mzoughi, H.; Njeh, I.; Wali, A.; Slima, M.B.; BenHamida, A.; Mhiri, C.; Mahfoudhe, K.B. Deep Multi-Scale
3D Convolutional Neural Network (CNN) for MRI Gliomas Brain Tumor Classification. J. Digit. Imaging
2020, 33, 903–915. [CrossRef] 22. Mzoughi, H.; Njeh, I.; Wali, A.; Slima, M.B.; BenHamida, A.; Mhiri, C.; Mahfoudhe, K.B. Deep Multi-Scale
3D Convolutional Neural Network (CNN) for MRI Gliomas Brain Tumor Classification. J. Digit. Imaging
2020, 33, 903–915. [CrossRef] 3. Siegel, R.L.; Miller, K.D.; Jemal, A. Cancer statistics, 2020. CA Cancer J. Clin. 2020, 70, 7–30. [CrossRef] 23. Siegel, R.L.; Miller, K.D.; Jemal, A. Cancer statistics, 2020. CA Cancer J. Clin. 2020, 70, 7–30. [CrossRef]
24. Amin, Z.S.; Fornaciari, E.; Ebert, L.M.; Koszyca, B.; Gomez, G.A. DeepSurvNet: Deep survival convolutional
network for brain cancer survival rate classification based on histopathological images. Med. Biol. Eng. Comput. 2020, 58, 1031–1045. 25. Perrin, S.L.;
Samuel, M.S.;
Koszyca, B.;
Brown, M.P.;
Ebert, L.M.;
Oksdath, M.;
Gomez, G.A. Glioblastoma heterogeneity and the tumour microenvironment: Implications for preclinical research
and development of new treatments. Biochem. Soc. Trans. 2019, 47, 625–638. [CrossRef] [PubMed] 26. Muhammad, K.; Khan, S.; Ser, J.D.; de Albuquerque, V.H.C. Deep learning for multigrade brain tumor
classification in smart healthcare systems: A prospective survey. IEEE Trans. Neural Netw. Learn. Syst. 2020. [CrossRef] [PubMed] 27. Szegedy, C.; Liu, W.; Jia, Y.; Sermanet, P.; Reed, S.; Anguelov, D.; Erhan, D.; Vanhoucke, V.; Rabinovich, A. Going deeper with convolutions. In Proceedings of the IEEE Conference on Computer Vision and Pattern
Recognition, Boston, MA, USA, 7–12 June 2015. 28. Liu, S.; Shah, Z.; Sav, A.; Russo, C.; Berkovsky, S.; Qian, Y.; Coiera, E.; Di Ieva, A. Isocitrate dehydrogenase
(iDH) status prediction in histopathology images of gliomas using deep learning. Sci. Rep. 2020, 10, 1–11. [CrossRef] [PubMed] 29. Sumi, P.S.; Delhibabu, R. Glioblastoma Multiforme Classification On High Resolution Histology
Image Using Deep Spatial Fusion Network. In Proceedings of the CEUR Workshop, Como, Italy,
9–11 September 2019; pp. 109–120. 30. Yonekura, A.; Kawanaka, H.; Prasath, V.S.; Aronow, B.J.; Takase, H. References Deep learning for drug response prediction in cancer. Brief. Bioinform. 2020. [CrossRef] 2. Shao, L.; Shum, H.P.; Hospedales, T. Special Issue on Machine Vision with Deep Learning. Int. J. Comput. 2020, 128, 771–772. [CrossRef] 13. Gour, M.; Jain, S.; Kumar, T.S. Residual learning based CNN for breast cancer histopathological image
classification. Int. J. Imaging Syst. Technol. 2020, 30, 621–635. [CrossRef] 14. Chen, X.; Men, K.; Chen, B.; Tang, Y.; Zhang, T.; Wang, S.; Li, Y.; Dai, J. CNN-Based Quality Assurance for
Automatic Segmentation of Breast Cancer in Radiotherapy. Front. Oncol. 2020, 10, 524. [CrossRef] [PubMed] 15. Duran-Lopez, L.; Dominguez-Morales, J.P.; Conde-Martin, A.F.; Vicente-Diaz, S.; Linares-Barranco, A. PROMETEO: A CNN-Based Computer-Aided Diagnosis System for WSI Prostate Cancer Detection. IEEE Access 2020, 8, 128613–128628. [CrossRef] 16. Liu, Z.; Yang, C.; Huang, J.; Liu, S.; Zhuo, Y.; Lu, X. Deep learning framework based on integration of
S-Mask R-CNN and Inception-v3 for ultrasound image-aided diagnosis of prostate cancer. Future Gener. Comput. Syst. 2020, 114, 358–367. [CrossRef] J. Pers. Med. 2020, 10, 224 24 of 27 17. Meng, J.; Xue, L.; Chang, Y.; Zhang, J.; Chang, S.; Liu, K.; Liu, S.; Wang, B.; Yang, K. Automatic detection and
segmentation of adenomatous colorectal polyps during colonoscopy using Mask R-CNN. Open Life Sci. 2020,
15, 588–596. [CrossRef] 18. Fonollà, R.; van der Zander, Q.E.; Schreuder, R.M.; Masclee, A.A.; Schoon, E.J.; van der Sommen, F. A CNN
CADx System for Multimodal Classification of Colorectal Polyps Combining WL, BLI, and LCI Modalities. Appl. Sci. 2020, 10, 5040. 19. Takahashi, M.; Kameya, Y.; Yamada, K.; Hotta, K.; Takahashi, T.; Sassa, N.; Iwano, S.; Yamamoto, T. An empirical study on the use of visual explanation in kidney cancer detection. In Proceedings of the
Twelfth International Conference on Digital Image Processing (ICDIP 2020), Osaka, Japan, 19–22 May 2020;
International Society for Optics and Photonics: Bellingham, WA, USA, 2020. 20. Vasanthselvakumar, R.; Balasubramanian, M.; Sathiya, S. Automatic Detection and Classification of Chronic
Kidney Diseases Using CNN Architecture. In Data Engineering and Communication Technology; Springer:
Berlin/Heidelberg, Germany, 2020; pp. 735–744. 21. Hashemzehi, R.; Mahdavi, S.J.S.; Kheirabadi, M.; Kamel, S.R. Detection of brain tumors from MRI images base
on deep learning using hybrid model CNN and NADE. Biocybern. Biomed. Eng. 2020, 40, 1225–1232. [CrossRef] 21. Hashemzehi, R.; Mahdavi, S.J.S.; Kheirabadi, M.; Kamel, S.R. Detection of brain tumors from MRI images base
on deep learning using hybrid model CNN and NADE. Biocybern. Biomed. Eng. References Automatic disease stage classification of
glioblastoma multiforme histopathological images using deep convolutional neural network. Biomed. Eng. Lett. 2018, 8, 321–327. [CrossRef] LeCun, Y.; Bottou, L.; Bengio, Y.; Haffner, P. Gradient-based learning applied to document recognition
Proc. IEEE 1998, 86, 2278–2324. [CrossRef] 32. Zeiler, M.D.; Fergus, R. Visualizing and understanding convolutional networks. In European Conference on
Computer Vision; Springer: Berlin/Heidelberg, Germany, 2014. 33. Simonyan, K.; Zisserman, A. Very deep convolutional networks for large-scale image recognition. arXiv
2014, arXiv:1409.1556. 34. Fukuma, R.; Yanagisawa, T.; Kinoshita, M.; Shinozaki, T.; Arita, H.; Kawaguchi, A.; Takahashi, M.; Narita, Y.;
Terakawa, Y.; Tsuyuguchi, N.; et al. Prediction of IDH and TERT promoter mutations in low-grade glioma
from magnetic resonance images using a convolutional neural network. Sci. Rep. 2019, 9, 1–8. [CrossRef] 35. Chang, K.; Bai, H.X.; Zhou, H.; Su, C.; Bi, W.L.; Agbodza, E.; Kavouridis, V.K.; Senders, J.T.; Boaro, A.;
Beers, A.; et al. Residual convolutional neural network for the determination of IDH status in low-and
high-grade gliomas from MR imaging. Clin. Cancer Res. 2018, 24, 1073–1081. [CrossRef] J. Pers. Med. 2020, 10, 224 25 of 27 36. Alqudah, A.M.; Alquraan, H.; Qasmieh, I.A.; Alqudah, A.; Al-Sharu, W. Brain Tumor Classification Using
Deep Learning Technique—A Comparison between Cropped, Uncropped, and Segmented Lesion Images
with Different Sizes. arXiv 2020, arXiv:2001.08844. [CrossRef] 37. Kalaiselvi, T.; Padmapriya, T.; Sriramakrishnan, P.; Priyadharshini, V. Development of automatic glioma
brain tumor detection system using deep convolutional neural networks. Int. J. Imaging Syst. Technol. 2020,
30, 926–938. [CrossRef] 38. Badža, M.M.; Barjaktarovi´c, M. ˇC. Classification of Brain Tumors from MRI Images Using a Convolutional
Neural Network. Appl. Sci. 2020, 10, 1999. [CrossRef] 39. Liu, D.; Liu, Y.; Dong, L. G-ResNet: Improved ResNet for brain tumor classification. In International Conference
on Neural Information Processing; Springer: Berlin/Heidelberg, Germany, 2019. 40. Hemanth, D.J.; Anitha, J.; Naaji, A.; Geman, O.; Popescu, D.E.; Son, L.H.; Hoang, L. A modified
deep convolutional neural network for abnormal brain image classification. IEEE Access 2018,
7, 4275–4283. [CrossRef] 41. Afshar, P.; Plataniotis, K.N.; Mohammadi, A. Capsule networks for brain tumor classification based on
MRI images and coarse tumor boundaries. In ICASSP 2019—2019 IEEE International Conference on Acoustics,
Speech and Signal Processing (ICASSP); IEEE: Piscataway Township, NJ, USA, 2019. Seetha, J.; Raja, S.S. Brain tumor classification using convolutional neural networks. Biomed. Pharmacol. J
2018, 11, 1457. [CrossRef] 43. Pashaei, A.; Sajedi, H.; Jazayeri, N. References Brain tumor classification via convolutional neural network and extreme
learning machines. In 2018 8th International Conference on Computer and Knowledge Engineering (ICCKE); IEEE:
Piscataway Township, NJ, USA, 2018. 44. Zhou, Y.; Li, Z.; Zhu, H.; Chen, C.; Gao, M.; Xu, K.; Xu, J. Holistic brain tumor screening and classification
based on densenet and recurrent neural network. In International MICCAI Brainlesion Workshop; Springer:
Berlin/Heidelberg, Germany, 2018. 45. Mohsen, H.; El-Dahshan, E.S.A.; El-Horbaty, E.S.M.; Salem, A.B.M. Classification using deep learning neural
networks for brain tumors. Future Comput. Inform. J. 2018, 3, 68–71. [CrossRef] 46. Ari, A.; Hanbay, D. Deep learning based brain tumor classification and detection system. Turk. J. Electr. Eng. Comput. Sci. 2018, 26, 2275–2286. [CrossRef] 47. Suter, Y.; Jungo, A.; Rebsamen, M.; Knecht, U.; Herrmann, E.; Wiest, R.; Reyes, M. Deep learning versus
classical regression for brain tumor patient survival prediction. In International MICCAI Brainlesion Workshop;
Springer: Berlin/Heidelberg, Germany, 2018. 48. Banerjee, S.; Mitra, S.; Masulli, F.; Rovetta, S. Brain tumor detection and classification from multi-sequence
MRI: Study using convnets. In International MICCAI Brainlesion Workshop; Springer: Berlin/Heidelberg,
Germany, 2018. 49. Afshar, P.; Mohammadi, A.; Plataniotis, K.N. Brain tumor type classification via capsule networks. In 2018
25th IEEE International Conference on Image Processing (ICIP); IEEE: Piscataway, NJ, USA, 2018. 50. Bagari, A.; Kumar, A.; Kori, A.; Khened, M.; Krishnamurthi, G. A combined Radio-Histological Approach
for Classification of Low Grade Gliomas. In International MICCAI Brainlesion Workshop; Springer:
Berlin/Heidelberg, Germany, 2018. g
y
51. Sajjad, M.; Khan, S.; Muhammad, K.; Wu, W.; Ullah, A.; Baik, S.W. Multi-grade brain tumor classification
using deep CNN with extensive data augmentation. J. Comput. Sci. 2019, 30, 174–182. [CrossRef] 52. Cheng, J.; Huang, W.; Cao, S.; Yang, R.; Yang, W.; Yun, Z.; Wang, Z.; Feng, Q. Enhanced performance of brain
tumor classification via tumor region augmentation and partition. PLoS ONE 2015, 10, e0140381. [CrossRef] 53. Clark, K.; Vendt, B.; Smith, K.; Freymann, J.; Kirby, J.; Koppel, P.; Moore, S.; Phillips, S.; Maffitt, D.;
Pringle, M.; et al. The Cancer Imaging Archive (TCIA): Maintaining and operating a public information
repository. J. Digit. Imaging 2013, 26, 1045–1057. [CrossRef] 54. Menze, B.H.; Jakab, A.; Bauer, S.; Kalpathy-Cramer, J.; Farahani, K.; Kirby, J.; Burren, Y.; Porz, N.; Slotboom, J.;
Wiest, R.; et al. The multimodal brain tumor image segmentation benchmark (BRATS). IEEE Trans. Med. Imaging 2014, 34, 1993–2024. [CrossRef] Vidoni, E.D. The Whole Brain Atlas: Www. med. harvard. edu/aanlib. J. Neurol. Phys. References Ther. 2012, 36, 108. 55. Vidoni, E.D. The Whole Brain Atlas: Www. m 56. Caldairou, B.; Passat, N.; Habas, P.A.; Studholme, C.; Rousseau, F. A non-local fuzzy segmentation method:
Application to brain MRI. Pattern Recognit. 2011, 44, 1916–1927. 56. Caldairou, B.; Passat, N.; Habas, P.A.; Studholme, C.; Rousseau, F. Application to brain MRI. Pattern Recognit. 2011, 44, 1916–1927. J. Pers. Med. 2020, 10, 224 26 of 27 26 of 27 57. Cancer Genome Atlas Research Network; Weinstein, J.N.; Collisson, E.A.; Mills, G.B.; Shaw, K.R.;
Ozenberger, B.A.; Ellrott, K.; Shmulevich, I.; Sander, C.; Stuart, J.M. The cancer genome atlas pan-cancer
analysis project. Nat. Genet. 2013, 45, 1113. [PubMed] 8. Brain Tumor Dataset. Available online:
https://figshare.com/articles/brain_tumor_dataset/15124
(accessed on 10 September 2020). p
59. BraTS2013. Available online: https://qtim-lab.github.io/ (accessed on 10 September 2020). 60. The Whole Brain Atlas. Available online:
http://www.med.harvard.edu/AANLIB/ (accessed on
10 September 2020). 61. BraTS2018. Available online:
https://www.med.upenn.edu/sbia/brats2018/data.html (accessed on
10 September 2020). 62. Devaki Scans & Diagnostics. Available online: https://www.medindia.net/labs/devaki-scans-diagnostics-
madurai-tamil-nadu-1680-1.htm (accessed on 10 September 2020). 63. Radiopaedia. Available online: https://radiopaedia.org/ (accessed on 10 September 2020). 64. BraTS2015. Available online: https://www.smir.ch/BRATS/Start2015 (accessed on 10 September 2020). 5. Suhag, S.; Saini, L.M. Automatic brain tumor detection and classification using svm classifier. of the ISER 2nd International Conference, Singapore, 20–21 September 2015. 6. Kwan, R.-S.; Evans, A.C.; Pike, G.B. MRI simulation-based evaluation of image-processing and classifica
methods. IEEE Trans. Med. Imaging 1999, 18, 1085–1097. [PubMed] 67. Scarpace, L.; Mikkelsen, L.; Cha, T.; Rao, S.; Tekchandani, S.; Gutman, S.; Pierce, D. Radiology data from the
cancer genome atlas glioblastoma multiforme [TCGA-GBM] collection. Cancer Imaging Arch. 2016, 11, 1. 68. Pedano, N.; Flanders, A.E.; Scarpace, L.; Mikkelsen, T.; Eschbacher, J.M.; Hermes, B.; Ostrom, Q. Radiology data from the cancer genome atlas low grade glioma [TCGA-LGG] collection. Cancer Imaging Arch. 2016, 2. 69. Ismael, S.A.A.; Mohammed, A.; Hefny, H. An enhanced deep learning approach for brain cancer MRI images
classification using residual networks. Artif. Intell. Med. 2020, 102, 101779. 70. Maharjan, S.; Alsadoon, A.; Prasad, P.W.C.; Al-Dalain, T.; Alsadoon, O.H. A novel enhanced softmax loss
function for brain tumour detection using deep learning. J. Neurosci. Methods 2020, 330, 108520. 71. Vijh, S.; Sharma, S.; Gaurav, P. Brain Tumor Segmentation Using OTSU Embedded Adaptive Particle Swarm
Optimization Method and Convolutional Neural Network. In Data Visualization and Knowledge Engineering;
Springer: Berlin/Heidelberg, Germany, 2020; pp. 171–194. 72. Rani, N.S.; Karthik, U.; Ranjith, S. References Extraction of Gliomas from 3D MRI Images using Convolution Kernel
Processing and Adaptive Thresholding. Procedia Comput. Sci. 2020, 167, 273–284. 73. Deng, W.; Shi, Q.; Wang, M.; Zheng, B.; Ning, N. Deep Learning-Based HCNN and CRF-RRNN Model for
Brain Tumor Segmentation. IEEE Access 2020, 8, 26665–26675. 74. Deng, W.; Shi, Q.; Luo, K.; Yang, Y.; Ning, N. Brain tumor segmentation based on improved convolutional
neural network in combination with non-quantifiable local texture feature. J. Med. Syst. 2019, 43, 152. [CrossRef] [PubMed] 75. Kumar, G.A.; Sridevi, P. Intensity Inhomogeneity Correction for Magnetic Resonance Imaging of
Automatic Brain Tumor Segmentation. In Microelectronics, Electromagnetics and Telecommunications; Springer:
Berlin/Heidelberg, Germany, 2019; pp. 703–711. 76. Mittal, M.; Goyal, L.M.; Kaur, S.; Kaur, I.; Verma, A.; Jude Hemanth, D. Hemanth Deep learning ba
enhanced tumor segmentation approach for MR brain images. Appl. Soft Comput. 2019, 78, 346–354. 77. Mittal, A.; Kumar, D. AiCNNs (Artificially-integrated Convolutional Neural Networks) for Brain Tu
Prediction. Eai Endorsed Trans. Pervasive Health Technol. 2019, 5, 346–354. [CrossRef] 78. Thillaikkarasi, R.; Saravanan, S. An enhancement of deep learning algorithm for brain tumor segmentation
using kernel based CNN with M-SVM. J. Med. Syst. 2019, 43, 84. [CrossRef] 79. Sharma, A.; Kumar, S.; Singh, S.N. Brain tumor segmentation using DE embedded OTSU method and neural
network. Multidimens. Syst. Signal Process. 2019, 30, 1263–1291. [CrossRef] 80. Kong, X.; Sun, G.; Wu, Q.; Liu, J.; Lin, F. Hybrid pyramid u-net model for brain tumor segmentation. In International Conference on Intelligent Information Processing; Springer: Berlin/Heidelberg Germany 2018 80. Kong, X.; Sun, G.; Wu, Q.; Liu, J.; Lin, F. Hybrid pyramid u-net model for brain tumor segmentation. In International Conference on Intelligent Information Processing; Springer: Berlin/Heidelberg, Germany, 2018. 81. Benson, E.; Pound, M.P.; French, A.P.; Jackson, A.S.; Pridmore, T.P. Deep hourglass for brain tumor
segmentation. In International MICCAI Brainlesion Workshop; Springer: Berlin/Heidelberg, Germany, 2018. J. Pers. Med. 2020, 10, 224 27 of 27 82. Zhou, C.; Chen, S.; Ding, C.; Tao, D. Learning contextual and attentive information for brain tumor
segmentation. In International MICCAI Brainlesion Workshop; Springer: Berlin/Heidelberg, Germany, 2018. 83. Dai, L.; Li, T.; Shu, H.; Zhong, L.; Shen, H.; Zhu, H. Automatic brain tumor segmentation with domain
adaptation. In International MICCAI Brainlesion Workshop; Springer: Berlin/Heidelberg, Germany, 2018. 84. Kermi, A.; Mahmoudi, I.; Khadir, M.T. Deep convolutional neural networks using U-Net for automatic brain
tumor segmentation in multimodal MRI volumes. References In International MICCAI Brainlesion Workshop; Springer:
Berlin/Heidelberg, Germany, 2018. g
y
85. Mlynarski, P.; Delingette, H.; Criminisi, A.; Ayache, N. Deep learning with mixed supervision for brain
tumor segmentation. J. Med. Imaging 2019, 6, 034002. [CrossRef] 86. Wang, G.; Li, W.; Zuluaga, M.A.; Pratt, R.; Patel, P.A.; Aertsen, M.; Doel, T.; David, A.L.; Deprest, J.;
Ourselin, S.; et al. Interactive medical image segmentation using deep learning with image-specific fine
tuning. IEEE Trans. Med. Imaging 2018, 37, 1562–1573. [CrossRef] 87. Monteiro, M.; Newcombe, V.F.; Mathieu, F.; Adatia, K.; Kamnitsas, K.; Ferrante, E.; Das, T.; Whitehouse, D.;
Rueckert, D.; Menon, D.K. Multiclass semantic segmentation and quantification of traumatic brain injury
lesions on head CT using deep learning: An algorithm development and multicentre validation study. Lancet Digit. Health 2020, 2, 314–322. [CrossRef] 88. Xu, Y.; Jia, Z.; Ai, Y.; Zhang, F.; Lai, M.; Eric, I.; Chang, C. Deep convolutional activation features for large
scale brain tumor histopathology image classification and segmentation. In 2015 IEEE International Conference
on Acoustics, Speech and Signal Processing (ICASSP); IEEE: Piscataway Township, NJ, USA, 2015. 89. Internet Brain Segmentation Repository. Available online: https://datamed.org/display-item.php?repository=
0058&id=590247f25152c6571cff8916&query= (accessed on 11 November 2020). 90. CENTER-TBI. Available online: https://www.center-tbi.eu/data (accessed on 10 September 2020). 91. CQ500 Dataset. Available online: http://headctstudy.qure.ai/dataset (accessed on 10 September 2020) 92. DICOM Image Sample Sets. Available online: https://www.osirix-viewer.com/resources/dicom-image-
library/ (accessed on 10 September 2020). 93. BraTS2017. Available online:
https://www.med.upenn.edu/sbia/brats2017/data.html (accessed on
10 September 2020). p
94. MICCAI 2014 Boston. Available online: http://miccai2014.org/ (accessed on 10 September 2020). 95. Wang, Q.; Hu, B.; Hu, X.; Kim, H.; Squatrito, M.; Scarpace, L.; Decarvalho, A.C.; Lyu, S.; Li, P.; Li, Y.; et al. Tumor evolution of glioma-intrinsic gene expression subtypes associates with immunological changes in the
microenvironment. Cancer Cell 2017, 32, 42–56.e6. Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional
affiliations. Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional
affiliations. © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/). © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/). © 2020 by the authors. Licensee MDPI, Basel, Switzerland. References This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/).
|
https://openalex.org/W2997685121
|
https://www.mdpi.com/1996-1073/13/1/264/pdf
|
English
| null |
Design Considerations of Series-Connected Devices Based LLC Converter
|
Energies
| 2,020
|
cc-by
| 12,379
|
Received: 30 November 2019; Accepted: 2 January 2020; Published: 5 January 2020
Received: 30 November 2019; Accepted: 02 January 2020; Published: date Abstract: This paper describes the design of a Series-Connected Device based on a fixed–frequency LLC
resonant converter (SCDLLC). Isolation of the dc-dc converter like the LLC resonant converter is used
for the stability of the high voltage system such as a solid-state-transformer (SST). The series-connected
devices driving method is one of the methods applicable to a high voltage system. When driving
series-connected devices, an auxiliary circuit for voltage balancing between series-connected devices
is required, which can be simply implemented using a passive element. In this paper, LLC converter
design with balancing circuits configured in parallel with a device is provided, and both the simulations
and experiments were performed. Abstract: This paper describes the design of a Series Connected Device based on a fixed frequency
LLC resonant converter (SCDLLC). Isolation of the dc-dc converter like the LLC resonant converter
is used for the stability of the high voltage system such as a solid-state-transformer (SST). The series
connected devices driving method is one of the methods applicable to a high voltage system. When
driving series-connected devices, an auxiliary circuit for voltage balancing between series
connected devices is required, which can be simply implemented using a passive element. In this
paper, LLC converter design with balancing circuits configured in parallel with a device is provided
and both the simulations and experiments were performed. Keywords:
solid-state-transformer (SST); isolation dc-dc converter; LLC resonant converter;
series-connected devices
Keywords: solid-state-transformer (SST); isolation dc-dc converter; LLC resonant converter; Series-
connected devices Design Considerations of Series-Connected Devic
Based LLC Converter
Design Considerations of Series-Connected Devi
Based LLC Converter * Correspondence: yhcho98@konkuk.ac.kr; Tel.: +82-10-6207-0431
* Correspondence: yhcho98@konkuk.ac.kr; Tel.: +82-10-6207-0431
Received: 30 November 2019; Accepted: 2 January 2020; Published: 5 January 2020
Received: 30 November 2019; Accepted: 02 January 2020; Published: date energies
Article
Design Considerations of Series-Connected Devices
Based LLC Converter
Dongkwan Yoon, Sungmin Lee and Younghoon Cho *
Department of Electrical Engineering, Konkuk University, Seoul 05029, Korea; asdf0462@konkuk.ac.kr (D.Y.);
lsm2429@konkuk.ac.kr (S.L.)
* Correspondence: yhcho98@konkuk.ac.kr; Tel.: +82-10-6207-0431
Received: 30 November 2019; Accepted: 2 January 2020; Published: 5 January 2020
Abstract: This paper describes the design of a Series-Connected Device based on a fixed–frequency LLC
resonant converter (SCDLLC). Isolation of the dc-dc converter like the LLC resonant converter is used
for the stability of the high voltage system such as a solid-state-transformer (SST). The series-connected
devices driving method is one of the methods applicable to a high voltage system. When driving
series-connected devices, an auxiliary circuit for voltage balancing between series-connected devices
is required, which can be simply implemented using a passive element. In this paper, LLC converter
design with balancing circuits configured in parallel with a device is provided, and both the simulations
and experiments were performed.
Keywords:
solid-state-transformer (SST); isolation dc-dc converter; LLC resonant converter;
series-connected devices
1 I t
d
ti
Article
Design Considerations of Series-Connected Devices
Based LLC Converter
Dongkwan Yoon, Sungmin Lee and Younghoon Cho *
Department of Electrical Engineering, Konkuk University, Seoul 05029, Korea; asdf0462@konkuk.ac.kr (D.Y.);
lsm2429@konkuk.ac.kr (S.L.)
* Correspondence: yhcho98@konkuk.ac.kr; Tel.: +82-10-6207-0431
Received: 30 November 2019; Accepted: 02 January 2020; Published: date
Abstract: This paper describes the design of a Series-Connected Device based on a fixed–frequency
LLC resonant converter (SCDLLC). Isolation of the dc-dc converter like the LLC resonant converte
is used for the stability of the high voltage system such as a solid-state-transformer (SST). The series
connected devices driving method is one of the methods applicable to a high voltage system. When
driving series-connected devices, an auxiliary circuit for voltage balancing between series
connected devices is required, which can be simply implemented using a passive element. In thi
paper, LLC converter design with balancing circuits configured in parallel with a device is provided
and both the simulations and experiments were performed.
Keywords: solid-state-transformer (SST); isolation dc-dc converter; LLC resonant converter; Series
connected devices
1. Introduction energies
Article
Design Considerations of Series-Connected Devices
Based LLC Converter
Dongkwan Yoon, Sungmin Lee and Younghoon Cho *
Department of Electrical Engineering, Konkuk University, Seoul 05029, Korea; asdf0462@konkuk.ac.kr (D.Y.);
lsm2429@konkuk.ac.kr (S.L.)
* Correspondence: yhcho98@konkuk.ac.kr; Tel.: +82-10-6207-0431
Received: 30 November 2019; Accepted: 2 January 2020; Published: 5 January 2020
Abstract: This paper describes the design of a Series-Connected Device based on a fixed–frequency LLC
resonant converter (SCDLLC). Isolation of the dc-dc converter like the LLC resonant converter is used
for the stability of the high voltage system such as a solid-state-transformer (SST). The series-connected
devices driving method is one of the methods applicable to a high voltage system. When driving
series-connected devices, an auxiliary circuit for voltage balancing between series-connected devices
is required, which can be simply implemented using a passive element. In this paper, LLC converter
design with balancing circuits configured in parallel with a device is provided, and both the simulations
and experiments were performed. Keywords:
solid-state-transformer (SST); isolation dc-dc converter; LLC resonant converter;
series-connected devices
1 I t
d
ti
Article
Design Considerations of Series-Connected Devices
Based LLC Converter
Dongkwan Yoon, Sungmin Lee and Younghoon Cho *
Department of Electrical Engineering, Konkuk University, Seoul 05029, Korea; asdf0462@konkuk.ac.kr (D.Y.);
lsm2429@konkuk.ac.kr (S.L.)
* Correspondence: yhcho98@konkuk.ac.kr; Tel.: +82-10-6207-0431
Received: 30 November 2019; Accepted: 02 January 2020; Published: date
Abstract: This paper describes the design of a Series-Connected Device based on a fixed–frequency
LLC resonant converter (SCDLLC). Isolation of the dc-dc converter like the LLC resonant converte
is used for the stability of the high voltage system such as a solid-state-transformer (SST). The series
connected devices driving method is one of the methods applicable to a high voltage system. When
driving series-connected devices, an auxiliary circuit for voltage balancing between series
connected devices is required, which can be simply implemented using a passive element. In thi
paper, LLC converter design with balancing circuits configured in parallel with a device is provided
and both the simulations and experiments were performed. Keywords: solid-state-transformer (SST); isolation dc-dc converter; LLC resonant converter; Series
connected devices
1. Introduction g
Energies 2020, 13, 264; doi:10.3390/en13010264 energies energies Energies 2020, 13, 264 Energies 2020, 13, 264 2 of 14 MMC is a structure that increases the front of the circuit by stacking the circuit in series, which
is relatively easy to expand, and has the advantage of reducing filter size by increasing the Power
Conversion System (PCS) voltage level. But, since there is a separated DC link voltage, it is necessary
to control and balance the separated DC link voltage. In addition, all individual devices should be
controlled as the number of modules increases, which requires a digital computing device with many
Pulse Width Modulation (PWM) channels. On the other hand, the series-connected devices behave like
a single device, so even if many devices are connected in series, no digital computing device with many
PWM channels is required. Furthermore, due to the series-connected devices, PCS does not have the
separated DC link voltage. No additional control method is needed for DC link voltage balancing. On
the other hand, because the output voltage level of series-connected devices based on PCS decreases,
the filter size is larger than the MMC method. In addition, there is an issue with a switch voltage
imbalance between the series-connected switches. According to References [9–16], there are various reasons for causing a voltage imbalance. The
first reason is the error of the gate driver. The gate driver is composed of various passive components
and semiconductor devices like Negative, Positive, Negative (NPN) and Positive, Negative, Positive
(PNP) transistors. Errors in devices of the gate driver cause an unbalance in the gate signal, which
causes an imbalance in the voltage across the series-connected devices. The second reason is the error
of parasitic components of the device itself. The error of the output capacitor of devices affects the
switching speed and causes a voltage imbalance. The third reason is the parasitic capacitor from gate
to ground of each device. To solve voltage imbalance, various studies have been conducted [9–16]. In Reference [9], Active
Gate Driver using Field Programmable Gate Array (FPGA) with RCD (Resistor, Capacitor and Diode)
snubbers across each device method for balancing was proposed and tested. In the documents, the
three-phase half-bridge inverter was built in 12-series connected Insulated Gage Bipolar Transistor
(IGBT) and verified at a rated current within ±10 kV DC-link. In Reference [12], an active gate driver
using a current mirror with a steady-state balancing resistor was proposed and performed. 1. Introduction
1. Introduction Recently, with new technologies such as smart grid and DC distribution, SST have emerged. SST
connects directly to the grid instead of the traditional transformers to perform a variety of roles such
as power factor correction and DC distribution. Therefore, SST should be able to cope with the high
voltage of the grid, so that the various studies can be conducted [1–8]. To cope with high voltage, a
multi-module converter (MMC) or a series-connected switching devices method has been studied. Figure 1 shows the difference between MMC and the series-connected device method. Recently, with new technologies such as smart grid and DC distribution, SST have emerged. SST
connects directly to the grid instead of the traditional transformers to perform a variety of roles such
as power factor correction and DC distribution. Therefore, SST should be able to cope with the high
voltage of the grid, so that the various studies can be conducted [1–8]. To cope with high voltage, a
multi-module converter (MMC) or a series-connected switching devices method has been studied. Figure 1 shows the difference between MMC and the series-connected device method. (a)
(b)
Figure 1. Comparison structure between MMC and the series-connected devices-based converter. (a)
MMC; (b) series-connected devices-based converter. Figure 1. Comparison structure between MMC and the series-connected devices-based converter. (a)
MMC; (b) series-connected devices-based converter. (a) (b) (a) (b) Figure 1. Comparison structure between MMC and the series-connected devices-based converter. (a
MMC; (b) series-connected devices-based converter. Figure 1. Comparison structure between MMC and the series-connected devices-based converter. (a)
MMC; (b) series-connected devices-based converter. p
j
g
www.mdpi.com/journal/energies Energies 2020, 13, 264; doi:10.3390/en13010264 2. Design of Series-Connected Devices Balancing Circuit
Figure 2 depicts the series-connected device with variou
Figure 2 depicts the series-connected device with variou Passive snubbers implemented by passive devices (C, RC, RCD, etc.) are used for voltage balancing
circuits. Voltage imbalance of devices is somewhat limited by placing the passive snubber circuit
between drain to source. The use of passive snubber circuits reduces the switching speed of devices
and the voltage imbalance due to a gate signal imbalance. Figure 3 shows series-connected devices
with passive balancing circuits. The snubber circuit composed of a steady-state balancing resistor (Rbal)
and a transient state balancing capacitor (Csnub, Rdamp). Passive snubbers implemented by passive devices (C, RC, RCD, etc.) are used for voltage
balancing circuits. Voltage imbalance of devices is somewhat limited by placing the passive snubber
circuit between drain to source. The use of passive snubber circuits reduces the switching speed of
devices and the voltage imbalance due to a gate signal imbalance. Figure 3 shows series-connected
devices with passive balancing circuits. The snubber circuit composed of a steady-state balancing
resistor (Rbal) and a transient state balancing capacitor (Csnub, Rdamp). Figure 3. Structure of voltage balancing circuits in series-connected devices
Gate
Driver
Cdg
Cgs
Cds
Gate
Driver
Csnub
Rdamp
Rbal
Figure 3. Structure of voltage balancing circuits in series-connected devices. Gate
Driver
Cdg
Cgs
Cds
Gate
Driver
Csnub
Rdamp
Rbal
Figure 3. Structure of voltage balancing circuits in series-connected devices. Figure 3 Structure of voltage balancing circuits in series-connected devices
Figure 3. Structure of voltage balancing circuits in series-connected devices. Figure 3. Structure of voltage balancing circuits in series-connected devices. The steady-state balancing resistors operate when the switch is fully turned-off. If the steady-
state balancing resistors are small, the devices are well balanced. However, the power dissipation of
balancing resistors increased, which requires the use of higher-rated resistors. Conversely, If the
balancing resistors are big, the power dissipation of balancing resistors decrease. However, there is a
high probability that voltage balancing between series-connected devices will fail. Therefore, when
designing the balancing resistor, the leakage current of the device should be considered. Steady-state
i t
l
l t d b l
The steady-state balancing resistors operate when the switch is fully turned-off. If the steady-
state balancing resistors are small, the devices are well balanced. However, the power dissipation of
balancing resistors increased, which requires the use of higher-rated resistors. Conversely, If the
balancing resistors are big, the power dissipation of balancing resistors decrease. 2. Design of Series-Connected Devices Balancing Circuit
Figure 2 depicts the series-connected device with variou
Figure 2 depicts the series-connected device with variou Figure 2 depicts the series-connected device with various factors that cause a voltage imbalance. Gate signal mismatch can occur due to an error of the device’s parasitic capacitor, unbalance of the gate
signal due to a gate pattern, and an error of the parasitic capacitor from gate to ground, which causes
an unbalance of voltage across the device. Furthermore, the voltage imbalance of devices increases the
stress and can cause damage to the device. In this section, in order to prevent damage to the device,
the voltage balancing method of the series-connected devices was analyzed using passive components. Gate signal mismatch can occur due to an error of the device’s parasitic capacitor, unbalance of the
gate signal due to a gate pattern, and an error of the parasitic capacitor from gate to ground, which
causes an unbalance of voltage across the device. Furthermore, the voltage imbalance of devices
increases the stress and can cause damage to the device. In this section, in order to prevent damage
to the device, the voltage balancing method of the series-connected devices was analyzed using
passive components. Gate signal mismatch can occur due to an error of the device s parasitic capacitor, unbalance of the
gate signal due to a gate pattern, and an error of the parasitic capacitor from gate to ground, which
causes an unbalance of voltage across the device. Furthermore, the voltage imbalance of devices
increases the stress and can cause damage to the device. In this section, in order to prevent damage
to the device, the voltage balancing method of the series-connected devices was analyzed using
passive components. Figure 2. Voltage imbalance factors in series-connected devices. Figure 2. Voltage imbalance factors in series-connected devices. Figure 2. Voltage imbalance factors in series-connected devices. Figure 2. Voltage imbalance factors in series-connected devices. Figure 2. Voltage imbalance factors in series-connected devices. Figure 2. Voltage imbalance factors in series-connected devices. Passive snubbers implemented by passive devices (C, RC, RCD, etc.) are used for voltage
balancing circuits. Voltage imbalance of devices is somewhat limited by placing the passive snubber
circuit between drain to source. The use of passive snubber circuits reduces the switching speed of
devices and the voltage imbalance due to a gate signal imbalance. Figure 3 shows series-connected
devices with passive balancing circuits. The snubber circuit composed of a steady-state balancing
resistor (Rbal) and a transient state balancing capacitor (Csnub, Rdamp). Energies 2020, 13, 264 The
steady-state voltage imbalance is reduced by the balancing resistor, and the transient imbalance was
solved by the analog gate controller used a current mirror. In Reference [14], the voltage balancing
method with only passive components using a single gate driver unit was proposed. In the documents,
experimental verifications were applied to the DC circuit breaker with a 1.2 kV bus voltage. It was
confirmed that it is possible to cope with the high voltage system by using a series connection element
applied to various methods. In a high voltage system, like SST, isolated converters are needed to increase the stability of the
system and grid. Isolated converters are one of the necessary converters for SST regardless of MMC
or series-connected devices. There are many types of isolated converters such as Dual-Active-Bridge
(DAB), Quad-Active-Bridge (QAB), and LLC Converter. A fixed-frequency resonant converter conducts
zero voltage switching (ZVS) over a wide frequency range and provides the advantages of high efficiency
and high-power density. g
p
y
In this paper, fixed-frequency LLC resonant converter-based series-connected devices is proposed. In order to cope with a high voltage, a series-connected device based on PCS was applied and an R-C
snubber was applied to solve the voltage imbalance between series-connected devices. In conclusion,
this paper examined the effect of R-C snubber applied to the LLC converter to solve the voltage
imbalance. The proposed converter consists of a full-bridge inverter. Unlike the usual full-bridge
converter, the proposed converter consists of eight devices, as shown in Figure 1b. The operation of
the SCDLLC converter with a balancing circuit is described. This paper is structured as follows. In
Section 2, imbalance factors of series-connected devices are described and a balancing method using
passive elements is expressed. In Section 3, a configuration method of the LLC converter considering
series-connected devices is explained. In Sections 4 and 5, a 3-kW prototype model is manufactured
to verify the usefulness of the proposed system and the simulations and experiments are performed. Lastly, in Section 6, we discuss experimental results and conclude the paper. Energies 2020, 13, 264
Energies 2020, 13, x FOR
Energies 2020, 13, x FOR 3 of 14
3 of 15
3 of 15 w.
𝑣ௗ௦
ൈ𝑖_௫
൏𝑅൏
𝑣ௗ௦
10 ൈ𝑖_
(1)
𝑣ௗ௦
ൈ𝑖_௫
൏𝑅൏
𝑣ௗ௦
10 ൈ𝑖_
(1)
vds
10 × ileakage_max
< Rbal <
vds
10 × ileakage_min
(1) 3. Design of LLC Converter
g
Figure 4 depicts the ser Figure 4 depicts the series-connected SiC MOSFETs LLC Converter with voltage balancing circuits. Generally, the output capacitance of the switching device should be considered in the design process. However, in this case, Snubber capacitors are added in parallel with the series-connected devices for
voltage balancing. The output capacitor should be discharged during deadtime for a sufficient Zero
Voltage Switching (ZVS) effect. Therefore, the snubber capacitor should be considered when designing
an LLC converter. Except for the snubber capacitor, the others process of the designed LLC converter
are followed by the general LLC converter design process in References [17,18]. g
p
g
g
circuits. Generally, the output capacitance of the switching device should be considered in the design
process. However, in this case, Snubber capacitors are added in parallel with the series-connected
devices for voltage balancing. The output capacitor should be discharged during deadtime for a
sufficient Zero Voltage Switching (ZVS) effect. Therefore, the snubber capacitor should be considered
when designing an LLC converter. Except for the snubber capacitor, the others process of the
designed LLC converter are followed by the general LLC converter design process in References
[17,18]. Figure 4. Series-connected devices-based LLC Converter with a voltage balancing circuit. vpri
Lr_pri
Lm
Cr
Lr_sec
Np : Ns
Q1
Q4
Q2
Q3
iLr_pri
iLr_sec
+
-
vsec
+
-
+
-
Vo
+
-
Vin
Figure 4. Series-connected devices-based LLC Converter with a voltage balancing circuit. Figure 4. Series-connected devices-based LLC Converter with a voltage balancing circuit. Figure 4. Series-connected devices-based LLC Converter with a voltage balancing circuit. Figure 5 shows theoretical waveforms of the conventional LLC converter. For simplifying it to
analyze, some assumptions are made as follows. Figure 5 shows theoretical waveforms of the conventional LLC converter. For simplifying it to
analyze, some assumptions are made as follows. Figure 5 shows theoretical waveforms of the conventional LLC converter. For simplifying it to
analyze, some assumptions are made as follows. Figure 5 shows theoretical waveforms of the conventional LLC converter. For simplifying it to
analyze, some assumptions are made as follows. •
There is no error of parasitic elements of a switching device;
•
There is no error of parasitic elements of a switching device; •
Gate drivers are all ideal, and there is no time delay between series-connected devices due to the
gate driver. Energies 2020, 13, 264
The transient Energies 2020, 13, 264
The transient Energies 2020, 13, 264
The transient 4 of 14
te and The transient state balancing circuit (Csnub, Rdamp) operates when the switch is a turn-on state and a
turn-offstate. Csnub is selected considering the parasitic capacitor of the device. To avoid imbalance by
parasitic capacitor errors of the devices, attach a balancing capacitor with negligible parasitic capacitors
of a device. If snubber capacitance is big, voltage unbalance during the transient will be stabilized, but
switching speed will decrease and switching loss will increase. If the snubber capacitance is small, the
snubber capacitors cannot perform proper balancing. Rsnub works as a damping resistor. The inrush
current of the snubber capacitor at the switching state is limited by Rsnub. Therefore, Rsnub is selected
by the current rating of the snubber capacitor. In addition, when calculating the Rsnub value, a time
constant (τ = RC) and switching frequency (fsw) should be considered. A large time constant reduces
the switching speed, increases switching losses, and also hinders soft switching of the LLC converter. g
p
p
by parasitic capacitor errors of the devices, attach a balancing capacitor with negligible parasitic
capacitors of a device. If snubber capacitance is big, voltage unbalance during the transient will be
stabilized, but switching speed will decrease and switching loss will increase. If the snubber
capacitance is small, the snubber capacitors cannot perform proper balancing. Rsnub works as a
damping resistor. The inrush current of the snubber capacitor at the switching state is limited by Rsnub. Therefore, Rsnub is selected by the current rating of the snubber capacitor. In addition, when calculating
the Rsnub value, a time constant (𝜏= 𝑅𝐶) and switching frequency (fsw) should be considered. A large
time constant reduces the switching speed, increases switching losses, and also hinders soft switching
of the LLC converter. 3 Design of LLC Converter 2. Design of Series-Connected Devices Balancing Circuit
Figure 2 depicts the series-connected device with variou
Figure 2 depicts the series-connected device with variou However, there is a
high probability that voltage balancing between series-connected devices will fail. Therefore, when
designing the balancing resistor, the leakage current of the device should be considered. Steady-state
resistance are calculated below. The steady-state balancing resistors operate when the switch is fully turned-off. If the steady-state
balancing resistors are small, the devices are well balanced. However, the power dissipation of
balancing resistors increased, which requires the use of higher-rated resistors. Conversely, If the
balancing resistors are big, the power dissipation of balancing resistors decrease. However, there is a
high probability that voltage balancing between series-connected devices will fail. Therefore, when
designing the balancing resistor, the leakage current of the device should be considered. Steady-state
resistance are calculated below. (1)
(1)
(1) Energies 2020, 13, 264 Energies 2020, 13, 264 Energies 2020, 13, 264 Equation (2) represents that the ZVS is related to the magnetizing inductance of the high
frequency transformer (HF transformer) and the output capacitance of the switch. As mentioned in the
previous section, Snubber capacitors are required for voltage balance between series-connected devices. Therefore, when manufacturing the LLC converter with balancing circuits that are connected in parallel
to each device, balancing circuits should be considered. This is not a parasitic component of the switch. After that, the rest process design of an LLC converter follows the conventional design process. Energies 2020, 13, x FOR PEER REVIEW
5 of 15 Fi
5 S
i hi
d
(i
)
i i
(i
) f h
f Figure 5. Switching state and resonant (iLr_pri), magnetizing current (iLm) of the transformer. Figure 5. Switching state and resonant (iLr_pri), magnetizing current (iLm) of the transformer. The gate signals of devices are off during the deadtime (tdead) interval. The magnetizing current
and resonant current become the same during the deadtime During the deadtime inductor current
Figure 6 shows the equivalent circuit of an LLC converter. To design LLC, the converter is
described as follows [3–9]. The gate signals of devices are off during the deadtime (tdead) interval. The magnetizing current
and resonant current become the same during the deadtime. During the deadtime, inductor current
Figure 6 shows the equivalent circuit of an LLC converter. To design LLC, the converter is
described as follows [3–9]. Np me during the deadtime. During the deadtime, inductor current
of the switch and discharges the parasitic output capacitor of the
er zero voltage is formed by the discharged output capacitor so
quation of minimum deadtime and magnetizing inductance for
he ZVS is related to the magnetizing inductance of the high
mer) and the output capacitance of the switch. As mentioned in
tors are required for voltage balance between series-connected
ring the LLC converter with balancing circuits that are connected
circuits should be considered. 3. Design of LLC Converter
g
Figure 4 depicts the ser •
Gate drivers are all ideal, and there is no time delay between series-connected devices due to the
gate driver. The gate signals of devices are offduring the deadtime (tdead) interval. The magnetizing current
and resonant current become the same during the deadtime. During the deadtime, inductor current
flows through the antiparallel diode of the switch and discharges the parasitic output capacitor of the
switch. Then, the switch turns on after zero voltage is formed by the discharged output capacitor so
that soft switching is possible. The equation of minimum deadtime and magnetizing inductance for
soft switching is calculated below. (2) tdead ≥16Coss fswLm
(2) tdead ≥16Coss fswLm 5 of 14 Energies 2020, 13, 264 This is not a parasitic component
cess design of an LLC converter follows the conventional design
𝑡ௗௗ16𝐶௦௦𝑓௦௪𝐿
(2)
n =
Np
Ns
(3)
m = Lr+Lm
Lr
(4)
fr =
1
2π
√
LrCr
(5)
Fx =
fsw
fr
(6)
Rac, min =
8
π2 ·
N2
p
N2s
·Ro
(7)
Qmax =
√
Lr/Cr
Rac,min
(8) f the
(3) high
d
(6) cted
cted
(7) esign
(8) Figure 6 shows the equivalent circuit of an LLC converter. To design LLC, the converter is
described as follows [3–9]. Np
where Np and Ns means the turn ratio of the transformer and Ro is the load resistance. Furthermore,
fsw is the switching frequency, fr is the resonant frequency, and the Rac is the reflected load resistance. Therefore, the voltage gain of the resonant tank is defined below. n = Ns
(3)
m = Lr+Lm
Lr
(4)
K =
Vo_ac(s)
Vinac(s)
=
F2
x(m −1)
q
m·F2x −1
2 + Fx2
F2x −1
2(m −1)2Q2
(9) (9) fr =
1
2πඥLrCr
(5)
𝐹௫ = 𝑓௦௪
𝑓
(6)
where Vo_ac (s), Vin_ac (s), Q, m, and Fx are the vin_ac, vo_ac, the quality factor, the ratio of the total primary
inductance to resonant inductance, and the normalized switching frequency, respectively. Figure 7
shows voltage gain according to different m and Q factors. In the figure, as the m factor decreases, the
voltage conversion ratio is extended. However, according to Equation (4), a low m factor requires low
magnetizing inductance Lm. Thus, magnetizing current increases. It means that the conduction loss of
magnetic components is increased. In terms of the Q factor, the Q factor is related to the load. The high 6 of 14
res low
on loss
n loss
d. The Energies 2020, 13, 264
voltage conversio
magnetizing indu
magnetizing induc
of magnetic compo Energies 2020, 13, 264
voltage conversio
magnetizing indu
magnetizing induc
of magnetic compo Q value means heavy load conditions and the low Q value means light load conditions. Therefore, it is
important to set the Q value at the rated load. Thus, a voltage gain of an LLC converter is related to m
and Q factors. high Q value means heavy load conditions and the low Q value means light load conditions. Therefore, it is important to set the Q value at the rated load. Figure 7. Voltage ga
4. Simulations and Simulation Results
4. Simulations and Simulation Results 4. Simulations and Simulation Results
In order to verify the series-connected devices-based LLC converter, simulation was conducted
. Si
u atio s a d Si
u atio
esu ts
In order to verify the series-connected devices-based LLC converter, simulation was conducted
by Power Sim (PSIM) and Linear Technology spice (LTspice). In order to verify the series-connected devices-based LLC converter, simulation was conducted by
Power Sim (PSIM) and Linear Technology spice (LTspice). Energies 2020, 13, 264 2.5
2.0
1.5
1.0
0.5
0.0
M = 3
1
Fx
2.5
2.0
1.5
1.0
0.5
0.0
M = 3
1
Fx Q = 0.4
2.5
2.0
1.5
1.0
0.5
0.0
M = 9
1
Fx
Q = 0.6
Q = 0.8
Q = 1.0
Q = 2.0
Q = 10.0
Q = 0.4
2.5
2.0
1.5
1.0
0.5
0.0
M = 9
1
Fx
Q = 0.6
Q = 0.8
Q = 1.0
Q = 2.0
Q = 10.0 2.5
2.0
1.5
1.0
0.5
0.0
M = 6
1
Fx
2.5
2.0
1.5
1.0
0.5
0.0
M = 6
1
Fx Fi
7 V lt
i
di
t diff
t
d Q f
t
Fx
F
Figure 7. Voltage gain according to different m and Q factors. Figure 7. Voltage gain according to different m and Q factors. Energies 2020, 13, 264 Thus, a voltage gain of an LLC converter
is related to m and Q factors. high Q value means heavy load conditions and the low Q value means light load conditions. Therefore, it is important to set the Q value at the rated load. Thus, a voltage gain of an LLC converter
is related to m and Q factors. Q value means heavy load conditions and the low Q value means light load conditions. Therefore, it is
important to set the Q value at the rated load. Thus, a voltage gain of an LLC converter is related to m
and Q factors. high Q value means heavy load conditions and the low Q value means light load conditions. Therefore, it is important to set the Q value at the rated load. Thus, a voltage gain of an LLC converter
is related to m and Q factors. g
Q
y
Q
g
Therefore, it is important to set the Q value at the rated load. Thus, a voltage gain of an LLC converter
is related to m and Q factors. Figure 6. Equivalent circuit of the LLC resonant tank. Figure 6. Equivalent circuit of the LLC resonant tank. Figure 6. Equivalent circuit of the LLC resonant tank. Figure 6. Equivalent circuit of the LLC resonant tank. Figure 6. Equivalent circuit of the LLC resonant tank. Figure 6. Equivalent circuit of the LLC resonant tank. Figure 7 Voltage gain according to different m and Q factors
Q = 0.4
2.5
2.0
1.5
1.0
0.5
0.0
2.5
2.0
1.5
1.0
0.5
0.0
2.5
2.0
1.5
1.0
0.5
0.0
M = 3
1
M = 6
M = 9
Fx
1
Fx
1
Fx
Q = 0.6
Q = 0.8
Q = 1.0
Q = 2.0
Q = 10.0
Figure 7. Voltage gain according to different m and Q factors. Q = 0.4
2.5
2.0
1.5
1.0
0.5
0.0
2.5
2.0
1.5
1.0
0.5
0.0
2.5
2.0
1.5
1.0
0.5
0.0
M = 3
1
M = 6
M = 9
Fx
1
Fx
1
Fx
Q = 0.6
Q = 0.8
Q = 1.0
Q = 2.0
Q = 10.0
Figure 7. Voltage gain according to different m and Q factors. by Power Sim (PSIM) and Linear
4 1 Voltage Balancing Simulation
4.1. Voltage Balancing Simulation 4.1. Voltage Balancing Simulation
Figure 8 shows the simulation circuit of voltage balancing. Table 1 shows the parasitic value of
components of experiment setup and Table 2 shows the parasitic value of device itself. Voltage
balancing simulation was performed by LTspice. Parasitic values of devices were added arbitrarily,
4.1. Voltage Balancing Simulation
Figure 8 shows the simulation circuit of voltage balancing. Table 1 shows the parasitic value of
components of experiment setup and Table 2 shows the parasitic value of device itself. Voltage
balancing simulation was performed by LTspice. Parasitic values of devices were added arbitrarily,
and voltage balancing simulation was performed depending on whether the snubber circuit was
Figure 8 shows the simulation circuit of voltage balancing. Table 1 shows the parasitic value
of components of experiment setup and Table 2 shows the parasitic value of device itself. Voltage
balancing simulation was performed by LTspice. Parasitic values of devices were added arbitrarily,
and voltage balancing simulation was performed depending on whether the snubber circuit was used. ge balancing simulation was performed depending on whether the snub
g
p
p
g
Table 1. Parameters of the voltage balance test circuit. Parameters
Values
Input voltage (VIN)
1000 V
Gate driver parasitic inductance (Lg1)
1 pH
Gate driver parasitic inductance (Lg2)
1.2 pH
Parasitic output capacitance (Coss_1)
170 pF
Parasitic output capacitance (Coss_2)
100 pF
Inductor (L)
3 mH
Voltage balancing resistor (Rbal)
1 MΩ
Damping resistor (Rsnub)
5 Ω
Voltage balancing circuits capacitor (Csnub)
1 nF
Table 2. Parameter of the selected device (C2M0040120D). Parameters
Minimum
Type
Maximum
Leakage current
x
1 µA
100 µA
Output capacitance
150 pF simulation was performed depending on whether
Table 1. Parameters of the voltage balance test circuit. 7 of 14
15 Energies 2020, 13, 264
E
i
2020 13 Figure 8. Multi-pulse test circuit for voltage balancing simulation. Gate
Driver
Gate
Driver
vsw_2
+
-
vsw_1
+
-
iL
VIN
+
-
L
MCU
Lg1
Lg2
Coss_1
Coss_2
Figure 8. Multi-pulse test circuit for voltage balancing simulation. Figure 8. Multi-pulse test circuit for voltage balancing simulation. Figure 8. Multi-pulse test circuit for voltage balancing simulation. Table 1. Parameters of the voltage balance test circuit. Parameters
Values
Input voltage (VIN)
1000 V
Gate driver parasitic inductance (Lg1)
1 pH
G t d i
iti i d
t
(L )
1 2
H
Figure 9 shows the simulation results with and without balancing circuits. by Power Sim (PSIM) and Linear
4 1 Voltage Balancing Simulation
4.1. Voltage Balancing Simulation Without balancing
circuits, simulation results show that the voltage balancing between series devices did not converge,
and the voltage imbalance increased as the inductor current increased. On the other hand, with
balancing circuits, voltage imbalance is somewhat reduced, and the imbalance range is maintained
even with the increased inductor current. Energies 2020, 13, x FOR PEER REVIEW
8 of 15 Gate driver parasitic inductance (Lg2)
1.2 pH
Parasitic output capacitance (Coss_1)
170 pF
Parasitic output capacitance (Coss_2)
100 pF
Inductor (L)
3 mH
Voltage balancing resistor (Rbal)
1 MΩ
Damping resistor (Rsnub)
5 Ω
Voltage balancing circuits capacitor (Csnub)
1 nF
Table 2. Parameter of the selected device (C2M0040120D). Parameters
Minimum
Type
Maximum
Leakage current
x
1 µA
100 µA
Output capacitance
150 pF
Figure 9 shows the simulation results with and without balancing circuits. Without balancing
circuits, simulation results show that the voltage balancing between series devices did not converge,
and the voltage imbalance increased as the inductor current increased. On the other hand, with
balancing circuits, voltage imbalance is somewhat reduced, and the imbalance range is maintained
even with the increased inductor current. (a)
(b)
Figure 9. Multi-pulse test simulation result: (a) without balancing circuits and (b) with balancing
circuits. 2 A
6 A
10 A
14 A
18 A
22 A
26 A
0
120 V
240 V
360 V
480 V
600 V
720 V
vsw_1
iL
vsw_2
2 A
6 A
10 A
14 A
18 A
22 A
26 A
0
120 V
240 V
360 V
480 V
600 V
720 V
vsw_1
vsw_2
iL
Figure 9. Multi-pulse test simulation result: (a) without balancing circuits and (b) with balancing circuits. 4.2. LLC Converter Simulation Parasitic output capacitance (Coss_1)
170 pF
Parasitic output capacitance (Coss_2)
100 pF
Inductor (L)
3 mH
Voltage balancing resistor (Rbal)
1 MΩ
Damping resistor (Rsnub)
5 Ω
Voltage balancing circuits capacitor (Csnub)
1 nF
Table 2. Parameter of the selected device (C2M0040120D). Parameters
Minimum
Type
Maximum
Leakage current
1
A
100
A
(a)
2 A
6 A
10 A
14 A
18 A
22 A
26 A
0
120 V
240 V
360 V
480 V
600 V
720 V
vsw_1
iL
vsw_2 Leakage current
x
1 µA
100 µA
Output capacitance
150 pF
re 9 shows the simulation results with and without balancing circuits. by Power Sim (PSIM) and Linear
4 1 Voltage Balancing Simulation
4.1. Voltage Balancing Simulation Without bal
imulation results show that the voltage balancing between series devices did not con
voltage imbalance increased as the inductor current increased. On the other hand
circuits, voltage imbalance is somewhat reduced, and the imbalance range is main
h the increased inductor current. (b)
2 A
6 A
10 A
14 A
18 A
22 A
26 A
0
120 V
240 V
360 V
480 V
600 V
720 V
vsw_1
vsw_2
iL Figure 9. Multi-pulse test simulation result: (a) without balancing circuits and (b) with balancing
circuits. 4 2 LLC C
Si
l i
Figure 9. Multi-pulse test simulation result: (a) without balancing circuits and (b) with balancing circuits. 4.2. LLC Converter Simulation 4.2. LLC Converter Simulation
The LLC converter simulation was conducted on the assumption that voltage imbalance did not
occur. The simulation model is shown in Figure 10. Table 3 gives the parameters of the system. An
isolated HF transformer can be described as an ideal transformer, magnetizing inductance, and
leakage inductance in simulation. In Figure 10, the model of the HF transformer of the LLC converter
consists of magnetizing leakage inductance and an ideal transformer, which has a 10:4 transfer ratio. Lm means the magnetizing inductance of the fabricated HF transformer. Lr_sec means the secondary
The LLC converter simulation was conducted on the assumption that voltage imbalance did
not occur. The simulation model is shown in Figure 10. Table 3 gives the parameters of the system. An isolated HF transformer can be described as an ideal transformer, magnetizing inductance, and
leakage inductance in simulation. In Figure 10, the model of the HF transformer of the LLC converter
consists of magnetizing leakage inductance and an ideal transformer, which has a 10:4 transfer ratio. Lm means the magnetizing inductance of the fabricated HF transformer. Lr_sec means the secondary 8 of 14
nd
er Energies 2020, 13, 264
leakage induc side leakage inductance of the HF transformer. Lr_pri means the primary side leakage inductance of the
HF transformer, which is the sum of the leakage inductance of the HF transformer itself and the added
inductance on the outside. In addition, Cr means the resonant capacitor. Resonant capacitors work as a
resonant tank with Lr_pri. Therefore, the resonant frequency of the LLC converter is determined by the
resonant capacitor and the primary leakage inductance of the transformer. by Power Sim (PSIM) and Linear
4 1 Voltage Balancing Simulation
4.1. Voltage Balancing Simulation Figure 11 shows the voltage
gain curve according to the load of the proposed converter. Lm means the magnetizing inductance of the fabricated HF transformer. Lr_sec means the secondary
side leakage inductance of the HF transformer. Lr_pri means the primary side leakage inductance of
the HF transformer, which is the sum of the leakage inductance of the HF transformer itself and the
added inductance on the outside. In addition, Cr means the resonant capacitor. Resonant capacitors
work as a resonant tank with Lr_pri. Therefore, the resonant frequency of the LLC converter is
determined by the resonant capacitor and the primary leakage inductance of the transformer. Figure
11 shows the voltage gain curve according to the load of the proposed converter. Figure 10. Simulation circuit of series-connected devices based on the LLC converter with a voltage
balancing circuit. Figure 10. Simulation circuit of series-connected devices based on the LLC converter with a voltage
balancing circuit. Table 3. Parameters of simulation. Table 3. Parameters of simulation. Parameters
Values
Input voltage (Vi)
1000 V
Output voltage (Vo)
400 V
Power
3000 W
Resonant frequency (fr)
100 kHz
Switching frequency (fsw)
100 kHz
HF transformer turn ratio (primary: secondary)
10:4
Primary leakage inductance (Lr_pri)
129 µH
Primary resonant capacitor (Cr)
20 nF
Magnetizing inductance (Lm)
302 µH
Secondary leakage inductance (Lr_sec)
2.598 µH
Primary resistance (Rpri)
0.135 Ω
Secondary resistance (Rsec)
0.110 Ω
Deadtime
500 ns Figures 12 and 13 show the simulation results of the LLC converter. Figure 12 shows the switching
voltage and channel current of the MOSFETs. In the figure, vds_Q1, vds_Q2, vds_Q3, and vds_Q4 represents
the drain to source voltages and iQ1, iQ2, iQ3, and iQ4 represents the current of devices. In the figure,
the channel currents increase after the devices are fully turned on. It means ZVS is achieved. Figure 13
shows the input/output voltage and the current of the HF transformer. Energies 2020, 13, x FOR PEER REVIEW
9 of 15
Figure 10. Simulation circuit of series-connected devices based on the LLC converter with a voltage
balancing circuit. K(Q, m, Fx)
Fx
K
14
12
10
8
6
4
2
0
100
16
500 W
1.0 kW
2.0 kW
2.5 kW
3.0 kW
1.5 kW Figure 11. Voltage gain graph of the LLC converter according to a different load. by Power Sim (PSIM) and Linear
4 1 Voltage Balancing Simulation
4.1. Voltage Balancing Simulation K(Q, m, Fx)
Fx
K
14
12
10
8
6
4
2
0
100
16
500 W
1.0 kW
2.0 kW
2.5 kW
3.0 kW
1.5 kW
Figure 11. Voltage gain graph of the LLC converter according to a different load. Figure 11. Voltage gain graph of the LLC converter according to a different load. Figure 11. Voltage gain graph of the LLC converter according to a different load. 9 of 14
f 15 Energies 2020, 13, 264
Energies 2020, 13, x F Figure 12. Switching waveform of the drain-source voltage and MOSFET current of the LLC
converter. (a)
(b)
(c)
(d)
(e)
(f)
Figure 13. Simulation results of the input voltage vpri and output voltage vsec of the transformer and
resonant current of the primary side iLr pri and secondary side iLr sec: (a) simulation at 500 W, (b)
600 V
400 V
200 V
0
600 V
400 V
200 V
0
600 V
400 V
200 V
0
600 V
400 V
200 V
0
30 A
20 A
10 A
0
30 A
20 A
10 A
0
30 A
20 A
10 A
0
30 A
20 A
10 A
0
ZVS
ZVS
ZVS
ZVS
ZVS
ZVS
ZVS
ZVS
vds_Q1
vds_Q4
vds_Q2
vds_Q3
iQ1
iQ4
iQ2
iQ3
1000 V
500 V
0
-1000 V
-500 V
15 A
7.5 A
vpri
vsec
vpri
vsec
iLr_pri
iLr_sec
iLr_pri
iLr_sec
ZVS
ZVS
ZVS
ZVS
0
-7.5 A
-15 A
1000 V
500 V
0
-1000 V
-500 V
vpri
vsec
vpri
vsec
iLr_pri
iLr_sec
iLr_pri
iLr_sec
ZVS
ZVS
ZVS
ZVS
15 A
7.5 A
0
-7.5 A
-15 A
1000 V
500 V
0
-1000 V
-500 V
vpri
vsec
vpri
vsec
iLr_pri
iLr_sec
iLr_pri
iLr_sec
ZVS
ZVS
ZVS
ZVS
15 A
7.5 A
0
-7.5 A
-15 A
Figure 12. Switching waveform of the drain-source voltage and MOSFET current of the LLC converter. Figure 12. Switching waveform of the drain-source voltage and MOSFET current of the LLC
converter. (a)
(b)
(c)
(d)
(e)
(f)
Figure 13. by Power Sim (PSIM) and Linear
4 1 Voltage Balancing Simulation
4.1. Voltage Balancing Simulation Simulation results of the input voltage vpri and output voltage vsec of the transformer and
resonant current of the primary side iLr_pri and secondary side iLr_sec: (a) simulation at 500 W, (b)
600 V
400 V
200 V
0
600 V
400 V
200 V
0
600 V
400 V
200 V
0
600 V
400 V
200 V
0
30 A
20 A
10 A
0
30 A
20 A
10 A
0
30 A
20 A
10 A
0
30 A
20 A
10 A
0
ZVS
ZVS
ZVS
ZVS
ZVS
ZVS
ZVS
ZVS
vds_Q1
vds_Q4
vds_Q2
vds_Q3
iQ1
iQ4
iQ2
iQ3
1000 V
500 V
0
-1000 V
-500 V
15 A
7.5 A
vpri
vsec
vpri
vsec
iLr_pri
iLr_sec
iLr_pri
iLr_sec
ZVS
ZVS
ZVS
ZVS
0
-7.5 A
-15 A
1000 V
500 V
0
-1000 V
-500 V
vpri
vsec
vpri
vsec
iLr_pri
iLr_sec
iLr_pri
iLr_sec
ZVS
ZVS
ZVS
ZVS
15 A
7.5 A
0
-7.5 A
-15 A
1000 V
500 V
0
-1000 V
-500 V
vpri
vsec
vpri
vsec
iLr_pri
iLr_sec
iLr_pri
iLr_sec
ZVS
ZVS
ZVS
ZVS
15 A
7.5 A
0
-7.5 A
-15 A
Figure 13. Simulation results of the input voltage vpri and output voltage vsec of the transformer and
resonant current of the primary side iLr_pri and secondary side iLr_sec: (a) simulation at 500 W, (b)
simulation at 1000 W, (c) simulation at 1500 W, (d) simulation at 2000 W, (e) simulation at 2500 W, and Fi
12 S
i hi
f
f
h
d
i
l
d MOSFET
f
h
LLC
600 V
400 V
200 V
0
600 V
400 V
200 V
0
600 V
400 V
200 V
0
600 V
400 V
200 V
0
30 A
20 A
10 A
0
30 A
20 A
10 A
0
30 A
20 A
10 A
0
30 A
20 A
10 A
0
ZVS
ZVS
ZVS
ZVS
ZVS
ZVS
ZVS
ZVS
vds_Q1
vds_Q4
vds_Q2
vds_Q3
iQ1
iQ4
iQ2
iQ3
Figure 12. Switching waveform of the drain-source voltage and MOSFET current of the LLC
converter. by Power Sim (PSIM) and Linear
4 1 Voltage Balancing Simulation
4.1. Voltage Balancing Simulation 400 V
200 V
0
600 V
400 V
200 V
0
600 V
400 V
200 V
0
600 V
400 V
200 V
0
20 A
10 A
0
30 A
20 A
10 A
0
30 A
20 A
10 A
0
30 A
20 A
10 A
0
ZVS
ZVS
ZVS
ZVS
ZVS
ZVS
ZVS
ZVS
vds_Q1
vds_Q4
vds_Q2
vds_Q3
iQ1
iQ4
iQ2
iQ3 Figure 12. Switching waveform of the drain-source voltage and MOSFET current of the LLC
converter
Figure 12. Switching waveform of the drain-source voltage and MOSFET current of the LLC converter. converter. converter. (a)
(b)
(c)
(d)
(e)
(f)
Figure 13. Simulation results of the input voltage vpri and output voltage vsec of the transformer and
resonant current of the primary side iLr_pri and secondary side iLr_sec: (a) simulation at 500 W, (b)
1000 V
500 V
0
-1000 V
-500 V
15 A
7.5 A
vpri
vsec
vpri
vsec
iLr_pri
iLr_sec
iLr_pri
iLr_sec
ZVS
ZVS
ZVS
ZVS
0
-7.5 A
-15 A
1000 V
500 V
0
-1000 V
-500 V
vpri
vsec
vpri
vsec
iLr_pri
iLr_sec
iLr_pri
iLr_sec
ZVS
ZVS
ZVS
ZVS
15 A
7.5 A
0
-7.5 A
-15 A
1000 V
500 V
0
-1000 V
-500 V
vpri
vsec
vpri
vsec
iLr_pri
iLr_sec
iLr_pri
iLr_sec
ZVS
ZVS
ZVS
ZVS
15 A
7.5 A
0
-7.5 A
-15 A
(a)
(b)
(c)
(d)
(e)
(f)
Figure 13. Simulation results of the input voltage vpri and output voltage vsec of the transformer and
resonant current of the primary side iLr_pri and secondary side iLr_sec: (a) simulation at 500 W, (b)
1000 V
500 V
0
-1000 V
-500 V
15 A
7.5 A
vpri
vsec
vpri
vsec
iLr_pri
iLr_sec
iLr_pri
iLr_sec
ZVS
ZVS
ZVS
ZVS
0
-7.5 A
-15 A
1000 V
500 V
0
-1000 V
-500 V
vpri
vsec
vpri
vsec
iLr_pri
iLr_sec
iLr_pri
iLr_sec
ZVS
ZVS
ZVS
ZVS
15 A
7.5 A
0
-7.5 A
-15 A
1000 V
500 V
0
-1000 V
-500 V
vpri
vsec
vpri
vsec
iLr_pri
iLr_sec
iLr_pri
iLr_sec
ZVS
ZVS
ZVS
ZVS
15 A
7.5 A
0
-7.5 A
-15 A
Figure 13. 5.1. Voltage Balancing Experiment
5.1. Voltage Balancing Experiment
5.1. Voltage Balancing Experiment 5.1. Voltage Balancing Experiment
5.1. Voltage Balancing Experiment
5.1. Voltage Balancing Experimen g
g
p
Voltage imbalance was tested through a multi-pulse test. The experimental setup is shown in
Figure 8 The experiment parameters are represented in Table 1
Voltage imbalance was tested through a multi-pulse test. The experimental setup is shown in
Figure 8. The experiment parameters are represented in Table 1. Voltage imbalance was tested through a multi-pulse test. The experimental setup is shown in
Figure 8 The experiment parameters are represented in Table 1 Figure 8. The experiment parameters are represented in Table 1. Figure 15 shows the experimental results without balancing circuits and with balancing circuits. Compared with the simulation results, the trend tends to be different. In the simulation, the factor of
voltage imbalance reflects the unbalance of the device’s parasitic and the delay of the gate signal. However, according to Reference [12], the parasitic capacitor from gate to ground one of the
important factors contribute to a voltage imbalance. In addition, this factor was not reflected in the
simulation. Although practical experiments, with the parasitic capacitors considered from gate to
ground, the voltage imbalance converges to a steady state through snubber circuits selected by
simulation. It means that, if the unbalance voltage does not exceed the rated voltage of the switch,
the switch can be regarded as a stable operation with selected balancing circuits by simulation
Figure 15 shows the experimental results without balancing circuits and with balancing circuits. Compared with the simulation results, the trend tends to be different. In the simulation, the factor
of voltage imbalance reflects the unbalance of the device’s parasitic and the delay of the gate signal. However, according to Reference [12], the parasitic capacitor from gate to ground one of the important
factors contribute to a voltage imbalance. In addition, this factor was not reflected in the simulation. Although practical experiments, with the parasitic capacitors considered from gate to ground, the
voltage imbalance converges to a steady state through snubber circuits selected by simulation. It
means that, if the unbalance voltage does not exceed the rated voltage of the switch, the switch can be
regarded as a stable operation with selected balancing circuits by simulation. Figure 8. The experiment parameters are represented in Table 1. 5.1. Voltage Balancing Experiment
5.1. Voltage Balancing Experiment
5.1. Voltage Balancing Experiment Figure 15 shows the experimental results without balancing circuits and with balancing circuits
Compared with the simulation results, the trend tends to be different. In the simulation, the factor o
voltage imbalance reflects the unbalance of the device’s parasitic and the delay of the gate signal
However, according to Reference [12], the parasitic capacitor from gate to ground one of the
important factors contribute to a voltage imbalance. In addition, this factor was not reflected in the
simulation. Although practical experiments, with the parasitic capacitors considered from gate to
ground, the voltage imbalance converges to a steady state through snubber circuits selected by
simulation. It means that, if the unbalance voltage does not exceed the rated voltage of the switch
the switch can be regarded as a stable operation with selected balancing circuits by simulation. (a)
(b)
Figure 15. Experimental results of the voltage imbalance test: (a) without balancing circuits and (b)
with balancing circuits. 100 us/div
10A/div
200V/div
vsw_1
vsw_2
iL
100 us/div
10A/div
200V/div
vsw_1
vsw_2
iL
(a)
(b)
Figure 15. Experimental results of the voltage imbalance test: (a) without balancing circuits and (b)
with balancing circuits. 100 us/div
10A/div
200V/div
vsw_1
vsw_2
iL
100 us/div
10A/div
200V/div
vsw_1
vsw_2
iL
Figure 15. Experimental results of the voltage imbalance test: (a) without balancing circuits and (b)
with balancing circuits. (b)
100 us/div
10A/div
200V/div
vsw_1
vsw_2
iL
(b)
100 us/div
10A/div
200V/div
vsw_1
vsw_2
iL (a)
100 us/div
10A/div
200V/div
vsw_1
vsw_2
iL
(a)
100 us/div
10A/div
200V/div
vsw_1
vsw_2
iL b) (b) a) (a) Figure 15. Experimental results of the voltage imbalance test: (a) without balancing circuits and (b)
with balancing circuits. Figure 15. Experimental results of the voltage imbalance test: (a) without balancing circuits and (b
with balancing circuits. Figure 15. Experimental results of the voltage imbalance test: (a) without balancing circuits and (b)
with balancing circuits. by Power Sim (PSIM) and Linear
4 1 Voltage Balancing Simulation
4.1. Voltage Balancing Simulation Simulation results of the input voltage vpri and output voltage vsec of the transformer and
resonant current of the primary side iLr_pri and secondary side iLr_sec: (a) simulation at 500 W, (b)
simulation at 1000 W, (c) simulation at 1500 W, (d) simulation at 2000 W, (e) simulation at 2500 W, and
(f) simulation at 3000 W. Z
(a) ZVS(b) (c)
ZVS
-7.5 A
-15 A
1000 V
500 V
0
-1000 V
-500 V
vpri
vsec
iLr_pri
iLr_sec
ZVS
ZVS
15 A
7.5 A
0
-7.5 A
-15 A
(c)
(e)
1000 V
500 V
0
-1000 V
-500 V
vpri
vsec
iLr_pri
iLr_sec
ZVS
ZVS
15 A
7.5 A
0
-7.5 A
-15 A ZV
(c) ZVS
(d) VS(f) (e) (e)
(f)
Figure 13. Simulation results of the input voltage vpri and output voltage vsec of the transformer and
resonant current of the primary side iLr_pri and secondary side iLr_sec: (a) simulation at 500 W, (b)
Figure 13. Simulation results of the input voltage vpri and output voltage vsec of the transformer and
resonant current of the primary side iLr_pri and secondary side iLr_sec: (a) simulation at 500 W, (b)
Figure 13. Simulation results of the input voltage vpri and output voltage vsec of the transformer and
resonant current of the primary side iLr_pri and secondary side iLr_sec: (a) simulation at 500 W, (b)
simulation at 1000 W, (c) simulation at 1500 W, (d) simulation at 2000 W, (e) simulation at 2500 W, and
(f) simulation at 3000 W. 10 of 14 Energies 2020, 13, 264
and (f) simulat
and (f) simu 5. Experiments and Experimental Results
The proposed circuit verified by simu
The proposed circuit verified by sim The proposed circuit verified by simulation was tested through experiments. Figure 14 shows the
experimental configuration of the proposed LLC converter using a 1200-V SiC MOSFET (C2M0040120D,
Cree) and 1200-V SiC Schottky Diode (C4D20120D, Cree). Experimental specifications are shown in
Table 3. Before the verification of series-connected devices based on an LLC converter, a multi-pulse
test is conducted to verify voltage balancing between series-connected devices. the experimental configuration of the proposed LLC converter using a 1200-V SiC MOSFET
(C2M0040120D, Cree) and 1200-V SiC Schottky Diode (C4D20120D, Cree). Experimental
specifications are shown in Table 3. Before the verification of series-connected devices based on an
LLC converter, a multi-pulse test is conducted to verify voltage balancing between series-connected
devices. the experimental configuration of the proposed LLC converter using a 1200-V SiC MOSFET
(C2M0040120D, Cree) and 1200-V SiC Schottky Diode (C4D20120D, Cree). Experimental
specifications are shown in Table 3. Before the verification of series-connected devices based on an
LLC converter, a multi-pulse test is conducted to verify voltage balancing between series-connected
devices. Figure 14. Hardware configuration for a series-connected LLC converter. Series connected
Full Bridge Converter
Full Bridge Rectifier
HF Transformer
Leakage inductor
Resonant capacitor
Figure 14. Hardware configuration for a series-connected LLC converter. Figure 14. Hardware configuration for a series-connected LLC converter
Series connected
Full Bridge Converter
Full Bridge Rectifier
HF Transformer
Leakage inductor
Resonant capacitor Figure 14. Hardware configuration for a series-connected LLC converter. Figure 14. Hardware configuration for a series-connected LLC converter. Figure 14. Hardware configuration for a series-connected LLC converte 5.2. Double Pulse Test
5.2. Double Pulse Tes
5.2. Double Pulse Test As mentioned in the previous section, an added snubber circuit for voltage balancing slows down
switching speed and increases switching loss. In this section, a Double Pulse Test (DPT) was conducted 11 of 14
g slows
PT) was 11 of 14
g slows
PT) was Energies 2020, 13, 264
down switching s to analyze switching losses with and without snubber circuits. The experimental setup is shown in
Figure 16 and parameters are shown in Table 4. Balancing snubber parameters are shown in Table 1. shown in Figure 16 and parameters are shown in Table 4. Balancing snubber parameters are shown
in Table 1. (a)
(b)
Figure 16. Double pulse test circuits: (a) without balancing circuits and (b) with balancing circuits. Figure 16. Double pulse test circuits: (a) without balancing circuits and (b) with balancing circuits. ergies 2020, 13, x FOR PEER REVIEW
12 o
As mentioned in the previous section, an added snubber circuit for voltage balancing slo
wn switching speed and increases switching loss. In this section, a Double Pulse Test (DPT) w
nducted to analyze switching losses with and without snubber circuits. The experimental setu
own in Figure 16 and parameters are shown in Table 4. Balancing snubber parameters are sho
Table 1. (a)
rgies 2020, 13, x FOR PEER REVIEW
As mentioned in the previous section, an
wn switching speed and increases switching
nducted to analyze switching losses with and
own in Figure 16 and parameters are shown i
Table 1. (b)
12 of
d snubber circuit for voltage balancing slo
In this section, a Double Pulse Test (DPT) w
out snubber circuits. The experimental setup
le 4. Balancing snubber parameters are show (b) (a) Figure 16. Double pulse test circuits: (a) without balancing circuits and (b) with balancing circuits. Figure 16. Double pulse test circuits: (a) without balancing circuits and (b) with balancing circuits. Table 4. Parameters of the double pulse test. Table 4. Parameters of the double pulse test. Parameters
Values
Input voltage (VIN)
500 V
Inductor (L)
1.5 mH Table 4. Parameters of the double pulse test. Parameters
Values
Input voltage (VIN)
500 V
Inductor (L)
1.5 mH DPT was conducted when the DC-link is 500 V. Figure 17 shows DPT results. According to
Figure 17, both turn-on and turn-offlosses increased with snubber circuits. However, the proposed
LLC converter operates ZVS turn-on at all loads. The turn-on loss can be negligible. 5.2. Double Pulse Test
5.2. Double Pulse Tes
5.2. Double Pulse Test (a)
(b)
Figure 16. Double pulse test circuits: (a) without balancing circuits and (b) with balancing circuits. (a)
(b)
Figure 17. Switching loss comparison according to the switch current: (a) without balancing circuits
and (b) with balancing circuits. Table 4. Parameters of the double pulse test. Parameters
Values
Input voltage (VIN)
500 V
Inductor (L)
1.5 mH
DPT was conducted when the DC-link is 500 V. Figure 17 shows DPT results. According
Figure 17, both turn-on and turn-off losses increased with snubber circuits. However, the propose
LLC converter operates ZVS turn on at all loads The turn on loss can be negligible
(a)
(b)
Figure 17. Switching loss comparison according to the switch current: (a) without balancing circuits
and (b) with balancing circuits. Figure 17. Switching loss comparison according to the switch current: (a) without balancing circuits
and (b) with balancing circuits. (b)
the switch current: (a) without balancing circuits
the double pulse test. Values
VIN)
500 V
1.5 mH
(b) (a)
Figure 17. Switching loss comparison according
and (b) with balancing circuits. Table 4. Parameters
Paramet
Input voltag
Inductor
(a) (b) (a) DPT was conducted when the DC-link is 500 V. Figure 17 shows DPT results. According
gure 17, both turn-on and turn-off losses increased with snubber circuits. However, the propo
ll l
d
h
l
b
l
bl
Figure 17. Switching loss comparison according to the switch current: (a) without balancing circuits
and (b) with balancing circuits. Figure 17. Switching loss comparison according to the switch current: (a) without balancing circuits
and (b) with balancing circuits. LLC converter operates ZVS tu
T
5.3. LLC Converter Experiment 5.3. LLC Converter Experiment
Based on the balancing circuit’s value selected through simulations and a multi-pulse test, the
series-connected devices based on the LLC converter experiment was conducted. Figure 18 shows
the main waveforms of the proposed LLC converter according to the load. In the figure, vpri and vsec
represent input/output voltage of the HF transformer, iLr_pri and iLr_sec represent input/output current
Parameters
Values
Input voltage (VIN)
500 V
Inductor (L)
1.5 mH
DPT was conducted when the DC-link is 500 V. Figure 17 shows DPT results. According to
Fi
17 b
h
d
ff l
i
d
i h
bb
i
i
H
h
d
Based on the balancing circuit’s value selected through simulations and a multi-pulse test, the
series-connected devices based on the LLC converter experiment was conducted. Figure 18 shows
the main waveforms of the proposed LLC converter according to the load. In the figure, vpri and vsec
represent input/output voltage of the HF transformer, iLr_pri and iLr_sec represent input/output current
of the HF transformer, according to the load. As shown in the previous section, experimental results
were well matched with simulation results. Energies 2020, 13, 264
of the HF transfor
were well matche 12 of 14
l results 12 of 14
l results (a)
(b)
(c)
(d)
(e)
(f)
Figure 18. Experimental results of the input voltage vpri and output voltage vsec of transformer and
resonant current of the primary side iLr_pri and the secondary side iLr_sec. (a) Simulation at 500 W, (b)
simulation at 1000 W, (c) simulation at 1500 W, (d) simulation at 2000 W, (e) simulation at 2500 W,
and (f) simulation at 3000 W. Figure 18. Experimental results of the input voltage vpri and output voltage vsec of transformer and
resonant current of the primary side iLr_pri and the secondary side iLr_sec. (a) Simulation at 500 W, (b)
simulation at 1000 W, (c) simulation at 1500 W, (d) simulation at 2000 W, (e) simulation at 2500 W, and
(f) simulation at 3000 W. (b) (a) (b) (a) (d) (c) (d) (c) (f) (e) (f) (e) Figure 18. Experimental results of the input voltage vpri and output voltage vsec of transformer and
resonant current of the primary side iLr_pri and the secondary side iLr_sec. 6. Conclusions
6. Conclusions 6. Conclusions
This paper describes the design consideration of the fixed frequency LLC converter considered
balancing circuit of series-connected devices circuit. In the introduction, the reason for the imbalance
voltage across the series-connected devices was described and discussed the need of an LLC
This paper describes the design consideration of the fixed frequency LLC converter considered
balancing circuit of series-connected devices circuit. In the introduction, the reason for the imbalance
voltage across the series-connected devices was described and discussed the need of an LLC
converter The proposed converter consists of a series connected devices based LLC converter
This paper describes the design consideration of the fixed frequency LLC converter considered
balancing circuit of series-connected devices circuit. In the introduction, the reason for the imbalance
voltage across the series-connected devices was described and discussed the need of an LLC converter. The proposed converter consists of a series-connected devices-based LLC converter. g
converter. The proposed converter consists of a series-connected devices-based LLC converter. In order to verify the proposed converter, a series-connected devices-based voltage balancing
experiment and the series-connected devices-based LLC converter was performed. The snubber
circuit, which is the simplest method among the various methods for voltage balancing, was applied
and the switching loss comparison and voltage balancing test was performed according to snubber
circuits. Furthermore, based on the snubber circuits-designed simulation and experiments, 3 kW
series-connected devices-based LLC converter was fabricated and verified. Through the experiments,
the voltage balancing between series-connected devices and soft switching operation of the proposed
converter are verified. The efficiency of a fabricated converter was measured 95.12% at 3 kW load. When the series-connected based LLC converter is fabricated with balancing snubber circuits, it is
confirmed that the LLC converter design is necessary considering the balancing snubber, and the
converter. The proposed converter consists of a series-connected devices-based LLC converter. In order to verify the proposed converter, a series-connected devices-based voltage balancing
experiment and the series-connected devices-based LLC converter was performed. The snubber
circuit, which is the simplest method among the various methods for voltage balancing, was applied
and the switching loss comparison and voltage balancing test was performed according to snubber
circuits. Furthermore, based on the snubber circuits-designed simulation and experiments, 3 kW
series-connected devices-based LLC converter was fabricated and verified. Through the experiments,
the voltage balancing between series-connected devices and soft switching operation of the proposed
converter are verified. LLC converter operates ZVS tu
T
5.3. LLC Converter Experiment (a) Simulation at 500 W, (b)
simulation at 1000 W, (c) simulation at 1500 W, (d) simulation at 2000 W, (e) simulation at 2500 W,
and (f) simulation at 3000 W. Figure 18. Experimental results of the input voltage vpri and output voltage vsec of transformer and
resonant current of the primary side iLr_pri and the secondary side iLr_sec. (a) Simulation at 500 W, (b)
simulation at 1000 W, (c) simulation at 1500 W, (d) simulation at 2000 W, (e) simulation at 2500 W, and
(f) simulation at 3000 W. Figure 19 shows vqs_1, vqs_2, and vgs at the rated power. vqs_1, vqs_2 represent the drain to source
voltage of each device connected in series at the rated power. Voltage balancing between series-
connected devices did not diverge, and both switches connected in series had achieved ZVS operation. Figure 20 shows the efficiency of series-connected devices-based LLC converter. The maximum
efficiency was measured at the rated load. Figure 19 shows vqs_1, vqs_2, and vgs at the rated power. vqs_1, vqs_2 represent the drain to
source voltage of each device connected in series at the rated power. Voltage balancing between
series-connected devices did not diverge, and both switches connected in series had achieved ZVS
operation. Figure 20 shows the efficiency of series-connected devices-based LLC converter. The
maximum efficiency was measured at the rated load. 13 of 14
14
f 15
14 of 15 Energies 2020, 13, 264
E
i
2020 13
FO
Energies 2020, 13, x FO Figure 19. Experimental results of series-connected devices voltage vq3_1, vq3_2, and vgs at the rated load. 2 us/div
10A/div
200V/div
vq3_1
vq3_2
vgs
iLr_pri
Figure 19. Experimental results of series-connected devices voltage vq3_1, vq3_2, and vgs at the rated load. Figure 19. Experimental results of series-connected devices voltage vq3_1, vq3_2, and vgs at the rated load. 2 us/div
10A/div
200V/div
vq3_1
vq3_2
vgs
iLr_pri Figure 19. Experimental results of series-connected devices voltage vq3_1, vq3_2, and vgs at the rated load. Figure 19. Experimental results of series-connected devices voltage vq3_1, vq3_2, and vgs at the rated load. Figure 19. Experimental results of series-connected devices voltage vq3_1, vq3_2, and vgs at the rated load. Figure 20 Efficiency of series-connected devices-based LLC converter
96
94
92
90
88
86
84
82
80
Efficiency (%)
85.11
91.18
93.1
94.26
94.85
95.12
500
1000
1500
2000
2500
3000
Output power (W)
Figure 20. Efficiency of series-connected devices-based LLC converter. 6. Conclusions
6. Conclusions The efficiency of a fabricated converter was measured 95.12% at 3 kW load. When the series-connected based LLC converter is fabricated with balancing snubber circuits, it is
confirmed that the LLC converter design is necessary considering the balancing snubber, and the
experimental results confirm that the consideration is valid
In order to verify the proposed converter, a series-connected devices-based voltage balancing
experiment and the series-connected devices-based LLC converter was performed. The snubber
circuit, which is the simplest method among the various methods for voltage balancing, was applied
and the switching loss comparison and voltage balancing test was performed according to snubber
circuits. Furthermore, based on the snubber circuits-designed simulation and experiments, 3 kW
series-connected devices-based LLC converter was fabricated and verified. Through the experiments,
the voltage balancing between series-connected devices and soft switching operation of the proposed
converter are verified. The efficiency of a fabricated converter was measured 95.12% at 3 kW load. When the series-connected based LLC converter is fabricated with balancing snubber circuits, it is
confirmed that the LLC converter design is necessary considering the balancing snubber, and the
experimental results confirm that the consideration is valid. experimental results confirm that the consideration is valid. Author Contributions: D.Y. implemented the system and conducted the experiments. Y.C. managed the paper. Author Contributions: D.Y. implemented the system and conducted the experiments. Y.C. managed the paper. S.L. assisted the idea simulation and the paper writing. Author Contributions: D.Y. implemented the system and conducted the experiments. Y.C. managed the paper. S.L. assisted the idea simulation and the paper writing. All authors have read and agreed to the published version
of the manuscript. S.L. assisted the idea simulation and the paper writing. Funding: The Human Resources Program in Energy Technology of the Korea Institute of Energy Technology
Evaluation and Planning (KETEP) supported this work, which granted a financial resource from the Ministry of
Funding: The Human Resources Program in Energy Technology of the Korea Institute of Energy Technology
Evaluation and Planning (KETEP) supported this work, which granted a financial resource from the Ministry of
Trade, Industry & Energy, Republic of Korea. (NO. 20194030202370) and the Human Resources Program in
Funding: The Human Resources Program in Energy Technology of the Korea Institute of Energy Technology
Evaluation and Planning (KETEP) supported this work, which granted a financial resource from the Ministry of
Trade, Industry & Energy, Republic of Korea. (NO. LLC converter operates ZVS tu
T
5.3. LLC Converter Experiment 96
94
92
90
88
86
84
82
80
Efficiency (%)
85.11
91.18
93.1
94.26
94.85
95.12
500
1000
1500
2000
2500
3000
Output power (W)
Figure 20. Efficiency of series-connected devices-based LLC converter. Fi
20 Effi i
f
i
d d
i
b
d LLC
p
p
(
)
Figure 20. Efficiency of series-connected devices-based LLC converter. Figure 20. Efficiency of series-connected devices-based LLC converter. 6. Conclusions
6. Conclusions 20194030202370) and the Human Resources Program in Energy 14 of 14 Energies 2020, 13, 264 Technology of the Korea Institute of Energy Technology Evaluation and Planning (KETEP) granted financial
resource from the Ministry of Trade, Industry & Energy, Republic of Korea. (Grant No. 20174010201540). Technology of the Korea Institute of Energy Technology Evaluation and Planning (KETEP) granted financial
resource from the Ministry of Trade, Industry & Energy, Republic of Korea. (Grant No. 20174010201540). Conflicts of Interest: The authors declare no conflict of interest. Conflicts of Interest: The authors declare no conflict of interest. Conflicts of Interest: The authors declare no conflict of interest. References 1. Falcones, S.; Ayyanar, R.; Mao, X. A DC–DC Multiport-Converter-Based Solid-State Transformer Integrating
Distributed Generation and Storage. IEEE Trans. Power Electron. 2013, 28, 2192–2203. [CrossRef] Falcones, S.; Ayyanar, R.; Mao, X. A DC–DC Multiport-Converter-Based Solid-State Transformer Integrating 1. Falcones, S.; Ayyanar, R.; Mao, X. A DC–DC Multiport-Converter-Based Solid-State Transformer Integrating
Distributed Generation and Storage. IEEE Trans. Power Electron. 2013, 28, 2192–2203. [CrossRef] ,
;
yy
,
;
,
p
g
Distributed Generation and Storage. IEEE Trans. Power Electron. 2013, 28, 2192–2203. [CrossRef] 2. Fan, H.; Li, H. High-Frequency Transformer Isolated Bidirectional DC–DC Converter Modules with High
Efficiency Over Wide Load Range for 20 kVA Solid-State Transformer. IEEE Trans. Power Electron. 2011, 26,
3599–3608. [CrossRef] 2. Fan, H.; Li, H. High-Frequency Transformer Isolated Bidirectional DC–DC Converter Modules with High
Efficiency Over Wide Load Range for 20 kVA Solid-State Transformer. IEEE Trans. Power Electron. 2011, 26,
3599–3608. [CrossRef] 3. Huang, A.Q. Medium-Voltage Solid-State Transformer: Technology for a Smarter and Resilient Grid. IEEE
Ind. Electron. Mag. 2016, 10, 29–42. [CrossRef] 3. Huang, A.Q. Medium-Voltage Solid-State Transformer: Technology for a Smarter and Resilient Grid. IEEE
Ind. Electron. Mag. 2016, 10, 29–42. [CrossRef] 4. Madhusoodhanan, S.; Tripathi, A.; Patel, D.; Mainali, K.; Kadavelugu, A.; Hazra, S.; Bhattacharya, S.;
Hatua, K. Solid-State Transformer and MV Grid Tie Applications Enabled by 15 kV SiC IGBTs and 10 kV SiC
MOSFETs Based Multilevel Converters. IEEE Trans. Ind. Appl. 2015, 51, 3343–3360. [CrossRef] 4. Madhusoodhanan, S.; Tripathi, A.; Patel, D.; Mainali, K.; Kadavelugu, A.; Hazra, S.; Bhattacharya, S.;
Hatua, K. Solid-State Transformer and MV Grid Tie Applications Enabled by 15 kV SiC IGBTs and 10 kV SiC
MOSFETs Based Multilevel Converters. IEEE Trans. Ind. Appl. 2015, 51, 3343–3360. [CrossRef] 5. She, X.; Huang, A.Q.; Wang, G. 3-D Space Modulation with Voltage Balancing Capability for a Cascaded
Seven-Level Converter in a Solid-State Transformer. IEEE Trans. Power Electron. 2011, 26, 3778–3789. [CrossRef] 6. She, X.; Yu, X.; Wang, F.; Huang, A.Q. Design and Demonstration of a 3.6-kV–120-V/10-kVA Solid-State
Transformer for Smart Grid Application. IEEE Trans. Power Electron. 2014, 29, 3982–3996. [CrossRef] 7. Shi, J.; Gou, W.; Yuan, H.; Zhao, T.; Huang, A.Q. Research on voltage and power balance control for c
modular solid-state transformer. IEEE Trans. Power Electron. 2011, 26, 1154–1166. [CrossRef] 8. Zhao, T.; Wang, G.; Bhattacharya, S.; Huang, A.Q. Voltage and Power Balance Control for a Cascaded H
Converter-Based Solid-State Transformer. IEEE Trans. Power Electron. 2013, 28, 1523–1532. References [CrossRe ,
;
g,
;
y ,
;
g,
Q
g
g
nverter-Based Solid-State Transformer. IEEE Trans. Power Electron. 2013, 28, 1523–1532. [CrossRef] 9. Hou, K.; Zheng, Y.; Wang, X.; W, L.I.; He, A.; Jiang, Y. Practical Research on VSC Prototype Based on IGBTs
Serial Connected Technology. IEEE Trans. Power Electron. 2019, 34, 495–502. [CrossRef] 10. Ji, S.; Wang, F.; Tolbert, L.M.; Lu, T.; Zhao, Z.; Yu, H. An FPGA-Based Voltage Balancing Control for
Multi-HV-IGBTs in Series Connection. IEEE Trans. Ind. Appl. 2018, 54, 4640–4649. [CrossRef] 11. Ju Won, B.; Dong-Wook, Y.; Heung-Geun, K. High-voltage switch using series-connected IGBTs with simple
auxiliary circuit. IEEE Trans. Ind. Appl. 2001, 37, 1832–1839. [CrossRef] 12. Marzoughi, A.; Burgos, R.; Boroyevich, D. Active Gate-Driver with dv/dt Controller for Dynamic Voltage
Balancing in Series-Connected SiC MOSFETs. IEEE Trans. Ind. Electron. 2019, 66, 2488–2498. [CrossRef] 13. Raciti, A.; Belverde, G.; Galluzzo, A.; Greco, G.; Melito, M.; Musumeci, S. Control of the switching transients
of IGBT series strings by high-performance drive units. IEEE Trans. Ind. Electron. 2001, 48, 482–490. [CrossRef] 14. Ren, Y.; Yang, X.; Zhang, F.; Wang, K.; Chen, W.; Wang, L.; Pei, Y. A Compact Gate Control and
Voltage-Balancing Circuit for Series-Connected SiC MOSFETs and Its Application in a DC Breaker. IEEE
Trans. Ind. Electron. 2017, 64, 8299–8309. [CrossRef] 15. Sasagawa, K.; Abe, Y.; Matsuse, K. Voltage-balancing method for IGBTs connected in series. IEEE Trans. Ind. Appl. 2004, 40, 1025–1030. [CrossRef] 16. Zhang, F.; Yang, X.; Ren, Y.; Feng, L.; Chen, W.; Pei, Y. A Hybrid Active Gate Drive for Switching Loss
Reduction and Voltage Balancing of Series-Connected IGBTs. IEEE Trans. Power Electron. 2017, 32, 7469–7481. [CrossRef] 17. Rahman, S. Resonant LLC Converter: Operation and Design; Infineon Technologies North America (IFNA) Core:
Durham, NC, USA, 2012. 18. Hu, Z.; Wang, L.; Qiu, Y.; Liu, Y.; Sen, P.C. An Accurate Design Algorithm for LLC Resonant Converters—Part
II. IEEE Trans. Power Electron. 2016, 31, 5448–5460. [CrossRef] © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/).
|
https://openalex.org/W4367331863
|
https://ucl.scienceopen.com/document_file/fbcc26dc-e2e2-4751-bad3-3e9ded6b3018/ScienceOpen/ucloe-05-057.pdf
|
English
| null |
Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines: predicting future trends using a support vector machine algorithm and the Markov chain model
|
UCL Open environment
| 2,023
|
cc-by
| 17,196
|
Peer review UCL Open: Environment is an open scholarship publication, this article has been peer-reviewed through the
journal’s standard open peer review process. All previous versions of this article and open peer review reports
can be found online in the UCL Open: Environment Preprint server at ucl.scienceopen.com Copyright and open access
© 2023 The Authors. Creative Commons Attribution Licence (CC BY) 4.0 International licence
https://creativecommons.org/licenses/by/4.0/ Open access Open access
This is an open access article distributed under the terms of the Creative Commons Attribution Licence
(CC BY) 4.0 https://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, distribution and
reproduction in any medium, provided the original author and source are credited. *Corresponding author
E-mail: cris.cayetano@gmail.com
1College of Fisheries and Aquatic
Sciences, Western Philippines
University, Sta. Monica, Puerto
Princesa City, Palawan, Philippines
2Remote Sensing Group, Plymouth
Marine Laboratory, Prospect Place,
Plymouth PL4 7QP, UK Multi-spatiotemporal analysis of
changes in mangrove forests in
Palawan, Philippines: predicting
future trends using a support
vector machine algorithm and
the Markov chain model Cristobal B. Cayetano1* , Lota A. Creencia1 , Emma Sullivan2 , Daniel Clewley2
and Peter I. Miller2 Cristobal B. Cayetano1* , Lota A. Creencia1 , Emma Sullivan2 , Daniel Clewley2
and Peter I. Miller2 How to cite How to cite
Cayetano CB, Creencia LA, Sullivan E, Clewley D, Miller PI. Multi-spatiotemporal analysis of changes
in mangrove forests in Palawan, Philippines: predicting future trends using a support vector machine
algorithm and the Markov chain model. UCL Open: Environment. 2023;(5):04. Available from: https://doi. org/10.14324/111.444/ucloe.000057 Submission date: 4 July 2022; Acceptance date: 16 March 2023; Publication date: 28 April 2023 Introduction Mangroves are a group of complex trees and shrubs that naturally inhabit the intertidal zones of
coastal tropical and subtropical regions [1,2]. Although they can tolerate a wide range of salinity,
from hypersaline waters exceeding 100 parts per thousand to lower salinities of 2 parts per
thousand [3], they cannot compete reproductively with other terrestrial plants because the latter
have a better adaptation to a higher-elevation environment [4]. Mangrove forests are one of the
most important coastal ecosystems because they provide bio-productivity, for example, timber and
firewood, and they provide protection from natural hazards and regulation of natural phenomena,
for example, floods, storm erosion and salt intrusion [1,5,6]. They serve as a nursery and a habitat
for biodiversity, for example, breeding and spawning [7–9], they are of socio-economic and
cultural importance, for example, providing livelihoods, ecotourism, recreation and are of aesthetic
importance [10,11], and help mitigate climate change, for example, carbon sequestration [10,12]. There are about 65 mangrove species around the world [13], of which at least 50% currently
grow in the Philippines [14]. According to the Food and Agricultural Organization [15], Asia has
more extensive mangrove forests than any other continent. The Philippines is considered one of
the top biodiversity ‘hot spot’ countries in the world [16]. The Palawan Council for Sustainable
Development Staff (PCSDS) [17] initially reported 27 mangrove species in Palawan. About 22.23%
(56,261.3 ha) of the remaining mangrove forests in the Philippines are found in Palawan [18]. However, the ability of this ecosystem to colonise and maintain its spatial setting is increasingly
being affected by anthropogenic disturbances [19]. Consequently, mangrove forest cover in the
Philippines has decreased from approximately 500,000 ha in 1918 to about 120,000 ha by the end
of 1995 [20,21]. Dodd and Ong reported that the two main contributing factors for this decline
are overexploitation of raw product and coastal land use conversions (e.g., agriculture, residential
settlements, industrial and aquaculture) [21]. Although recent estimates from the Department of
Environment and Natural Resources (DENR) [22] suggest an increase in mangrove extent in 2003
(to 247,362 ha), this estimate is still much lower than the estimated area covered in the previous
century. Mangrove ecosystems form a complex structure (e.g., less accessible Rhizophora’s complex
bifurcated and looping root structures), and the technical skills required and cost associated with
taking forest samples make extensive in-situ sampling difficult. Abstract However, as this research did not capture the environmental factors
that may have influenced the changes in mangrove patterns, it is suggested adding cellular
automata in future Markovian mangrove modelling. Keywords: change detection, image classification, Landsat, land use/land cover, Markov chain model, spatial
dynamics, support vector machine Abstract *Corresponding author
E-mail: cris.cayetano@gmail.com
1College of Fisheries and Aquatic
Sciences, Western Philippines
University, Sta. Monica, Puerto
Princesa City, Palawan, Philippines
2Remote Sensing Group, Plymouth
Marine Laboratory, Prospect Place,
Plymouth PL4 7QP, UK Multi-temporal remote sensing imagery can be used to explore how mangrove assemblages are
changing over time and facilitate critical interventions for ecological sustainability and effective
management. This study aims to explore the spatial dynamics of mangrove extents in Palawan,
Philippines, specifically in Puerto Princesa City, Taytay and Aborlan, and facilitate future predictions
for Palawan using the Markov Chain model. The multi-date Landsat imageries during the period
1988–2020 were used for this research. The support vector machine algorithm was sufficiently
effective for mangrove feature extraction to generate satisfactory accuracy results (>70% kappa
coefficient values; 91% average overall accuracies). In Palawan, a 5.2% (2693 ha) decrease was
recorded during 1988–1998 and an 8.6% increase in 2013–2020 to 4371 ha. In Puerto Princesa
City, a 95.9% (2758 ha) increase was observed during 1988–1998 and 2.0% (136 ha) decrease
during 2013–2020. The mangroves in Taytay and Aborlan both gained an additional 2138 ha
(55.3%) and 228 ha (16.8%) during 1988–1998 but also decreased from 2013 to 2020 by 3.4%
(247 ha) and 0.2% (3 ha), respectively. However, projected results suggest that the mangrove 1College of Fisheries and Aquatic
Sciences, Western Philippines
University, Sta. Monica, Puerto
Princesa City, Palawan, Philippines 2Remote Sensing Group, Plymouth
Marine Laboratory, Prospect Place,
Plymouth PL4 7QP, UK 1 / 29 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
https://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
https://doi.org/10.14324/111.444/ucloe.000057 UCL OPEN ENVIRONMENT Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines areas in Palawan will likely increase in 2030 (to 64,946 ha) and 2050 (to 66,972 ha). This study
demonstrated the capability of the Markov chain model in the context of ecological sustainability
involving policy intervention. However, as this research did not capture the environmental factors
that may have influenced the changes in mangrove patterns, it is suggested adding cellular
automata in future Markovian mangrove modelling. areas in Palawan will likely increase in 2030 (to 64,946 ha) and 2050 (to 66,972 ha). This study
demonstrated the capability of the Markov chain model in the context of ecological sustainability
involving policy intervention. Study area Palawan is a long and narrow island province in the Philippines (09°30′N and 118°30′′E) with an
approximate total area of 1,489,626 ha and is located at the western portion of the archipelago
(Fig. 1) [17,45]. Its almost 2000 km coastline is one of the longest shorelines in the country and
accounts for about 1780 islands. The South China Sea borders the western coast while the Sulu
Sea and the Malaysian Sabah Island border the eastern and southern sides of Palawan [46]. The
island comprises 23 municipalities, one urbanised city (Puerto Princesa) and 433 small villages
called ‘barangay’ [47]. Palawan is known as the Philippines’ ‘last ecological frontier’ due to its rich culture and biodiversity
[48]. As per Presidential Proclamation No. 2152 of 1981, all mangrove forest areas in the province
are protected as Palawan has been declared a Mangrove Swamp Forest Reserve [17,45]. In 1991,
Palawan was designated as a biosphere reserve under the Man and the Biosphere Programme
(MAB) of the United Nations Educational, Scientific, and Cultural Organization (UNESCO). The
following year, the 1992 SEP Law assisted the MAB’s declaration in the sustainability of Palawan’s
biological and cultural diversity. In succeeding years of recognising the biodiversity richness of
the province, two out of nine UNESCO World Heritage Sites in the Philippines are to be found
in Palawan: the Puerto Princesa Subterranean River National Park (inscribed in 1999) and the
Tubbataha Reefs Natural Park (inscribed in 1993, 2009) [48]. Mangroves form one of the components of the coastal and marine ecosystems in the Philippines
[49]. They are susceptible to various effects of climate change such as sea-level rise [50]. Therefore,
adoption of various climate change adaptation interventions such as the National Framework
Strategy on Climate Change [51] and the development of the Philippine exposure map on climate
change [52] have been of great importance for the identification of vulnerable areas of Palawan that
are the most susceptible to climate change. The entire methodological process of mangrove classification and predictive modelling underwent
three major processes: (1) raw data and pre-processing; (2) image classification and change
detection; and (3) mangrove change projection (Fig. 2). classification algorithms can handle complex classification tasks [34]. To perform the classification
using a supervised classification technique, training samples must be extracted, which can be
time-consuming when using multi-temporal remotely sensed imagery. Unsupervised classification
techniques have also been used to map mangrove extent and change over time, for example, using
vegetation indices (e.g., the Normalised Difference Vegetation Index, the Mangrove Vegetation
Index [35,36]) and clustering and threshold techniques (e.g., [37]). The Markov chain model
[38,39] is one of many prediction techniques that are able to assess the LULC changes and make
a projection of these changes in the future [40–43]. Understanding the patterns of change in
mangrove geographic distribution and projecting the range of shifts in the future will link science to
policy and decision-making processes for biodiversity conservation and management [44]. Through the Global Challenges Research Fund (GCRF) Blue Communities (BC), this research aims
to: (1) develop a mapping approach to investigate the changes in mangrove extents in Palawan
using multi-temporal Landsat imagery during the years 1988, 1993, 1998, 2003, 2008, 2013, 2018
and 2020; (2) determine the areal extent of change in mangrove forests in Palawan including the
three case study areas of GCRF BC from 1988 to 2020; and (3) implement change projections of
the mangrove forests in Palawan for 2030 and 2050 using a Markov chain model. Introduction Thus, remote sensing techniques
provide a convenient tool to map, assess and monitor the mangroves over large areas and can
be used to detect change over time [23–25]. In the Philippines, the utilisation of remotely-sensed
satellite data (e.g., [18]) has been incorporated into policy formulation and enforcement. However,
mangrove-related projects in the country remain relatively scarce with only a few national and local
mapping efforts focused on the classification and detection of changes in the mangrove’s extent,
notably from the nominal years of 1990–2010 [26] and 2003–2013 [27]. Despite the low utilisation
of mangrove remote sensing in the Philippines and the absence of projected data about how
the remaining mangroves in the country will respond to the impacts of climate change, scientific
interest in mitigating and controlling the magnitude of climate change’s impacts on mangrove
ecosystems has increased in Southeast Asian countries [28]. The mangroves of Palawan have been
protected under direct human inventions through the International Union for Conservation of Nature
(IUCN) protected area Category I–IV [18] and the 1992 Republic Act No. 7611, commonly known
as the Strategic Environmental Plan for Palawan Act (SEP Law) [29]; yet this unique ecosystem
remains under threat due to climate change and the associated rising sea levels [18,30]. Several land use/land cover (LULC) techniques have been developed and utilised in the last
three decades, which primarily aim to investigate the spatiotemporal changes of LULC patterns
using satellite data to assist in ecological management and decision-making [31]. The parametric
(e.g., maximum likelihood classifier [32]) and nonparametric (e.g., artificial neural networks [33]) 2 / 29 UCL OPEN ENVIRONMENT Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
https://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
https://doi.org/10.14324/111.444/ucloe.000057 ttps://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Pre-processing the Landsat sensor data The multi-temporal resolution and multi-spectral Landsat 4–5 Thematic Mapper (TM), Landsat 7
Enhanced Thematic Mapper Plus (ETM+) and Landsat 8 Operational Land Imager (OLI) images in
multiple years between 1988 and 2020 were used for this study (Table A1 in Appendix A). A total of
20 scenes for TM (for years 1988, 1993 and 1998), 18 scenes for ETM+ (for years 2003, 2008 and
2013) and 11 scenes for OLI (for years 2018 and 2020) were sourced using the Semi-automatic
Classification Plugin (SCP) version 7.9.0 Matera in Quantum Geographical Information System
(QGIS) version 3.22.1 Białowiez˙a. 3 / 29 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
https://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
https://doi.org/10.14324/111.444/ucloe.000057 UCL OPEN ENVIRONMENT https://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Image classification
Spectral band selection
Cloud patching process
Datasets masking
Layer stacking and
mosaicking
Supervised SVM
implementation
LULC accuracy
validation
Change detection
analysis
Mangrove extraction of
the final maps
Image classification and
change detection
Geometric
correction
Noise correction
Conversion to
TOA units
Atmospheric
correction
Markov chain modelling
Markov computation
Transition probability
matrix
Transition area matrix
Markov chain projection
Model validation
Projected maps: 2013,
2030 and 2050
Landsat 4-5 TM,
Landsat 7 ETM+
and Landsat 8 OLI
Pre-Processing
Raw data and
pre-processing
Mangrove change
projection Figure 2 For the TM and ETM+ data,
Eqs (1)–(5) were applied, respectively: L
DN
G
B
λ =
×
+
(1) (1) L
DN
G
B
λ =
×
+ where Lλ corresponds to the radiance measured at the sensor bandwidth for each band
(W m−2 sr−1 μ−1); DN is the digital number value; G and B are the (gain) slope and (bias) intercept of
response functions, calculated as follows: max
min
min
(
/
)
B
L
L
L
Q
Q
Q
=
−
−
−
×
min
max
min
(2)
(
)
max
min
max
min
/
G
L
L
Q
Q
=
−
−
(3) max
min
min
(
/
)
B
L
L
L
Q
Q
Q
=
−
−
−
×
min
max
min
(2) (2)
(
)
max
min
max
min
/
G
L
L
Q
Q
=
−
−
(3) (
)
max
min
max
min
/
G
L
L
Q
Q
=
−
−
(3) (
)
max
min
max
min
/
G
L
L
Q
Q
=
−
−
(3) where Lmin and Lmax are the lowest and highest radiance measured by a detector in mW cm−2 sr−1, as
reported by TM and ETM+ metadata files; Qmin and Qmax correspond to the minimum and maximum
values of DN for TM and ETM+ sensors, ranging from 1 to 255. The TOA reflectance (ρλ) calculation
for each band applied on a pixel-by-pixel basis for each scene in each epoch and the output
reflectance values were scaled to an 8-bit data range, this can be calculated as: (
)
2
o
s
L
d
E
cos
π
ρ
θ
λ
λ
λ
×
×
=
×
(4) (
)
2
o
s
L
d
E
cos
π
ρ
θ
λ
λ
λ
×
×
=
×
(4) where d is the Earth–sun distance correction; Lλ is the radiance as a function of bandwidth; E0λ
is the mean solar exoatmospheric irradiances and θs is the solar zenith angle. The application of
absolute atmospheric correction and relative correction followed the corrections of sensor gains
and offsets spectral band solar irradiance and solar zenith angle, and the topographic normalisation
implementation. The removal of additive path radiance (Lp) was calculated using Eq. (5) based
on the dark-object subtraction (DOS) 1% technique [60–62]. Figure 2 Diagram of multi-temporal mangrove
change detection in Palawan using the
Landsat imageries, supervised support
vector machine classification and the
Markov chain model. Raw data and
pre-processing The 2018 OLI Level-2 data were used as the reference images to apply geometric correction to the
satellite images in each epoch. The parameters of this transformation function were derived from a
spread of 200 ground control points (GCPs), which were uniformly chosen from distinct topographic
features throughout the target image. To match with the original pixel size of the Landsat data,
all images were resampled to a ground resolution of 30 × 30 m and reprojected to WGS 84 UTM
datum. The root mean square error (RMSE) of 0.25 pixel was calculated and was deemed enough
to facilitate accurate LULC change detection analysis [57]. Throughout these processes, the
nearest neighbour resampling algorithm was employed to maintain geometric integrity across all the
images. Radiometric correction followed the geometric correction [55]. Upon checking the image noise
(e.g., dropouts and bit errors) for TM and ETM+ images using the Environmental Systems Research
Institute’s ArcGIS version 10.7.1, a correction was not necessary. The next process of radiometric
calibration involved the conversion of the signal of the quantified energy from multi-spectral
brightness values or digital numbers (DNs) into top-of-atmosphere (TOA) reflectance units. In
particular, this process involved two steps: (a) the conversion of DNs to spectral radiance (Lλ) and
(b) the transformation to TOA reflectance (ρλ) as corrected for illumination variabilities (i.e., sun angle
and Earth–sun distance) within and between scenes [55,56,58,59]. Figure 1 A map of Palawan, Philippines
highlighting the southern and northern
divisions and three of the GCRF BC’s
case study areas – Puerto Princesa City
and the municipalities of Taytay and
Aborlan. To normalise various conditions across the multi-temporal and multi-spatial Landsat datasets, it is
imperative that Landsat data undergoes pre-processing routines to enhance the quality and remove
various radiometric and geometric errors in each image [53–56]. Thus, radiometric calibration and
atmospheric correction were employed for this study. 4 / 29 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines UCL OPEN ENVIRONMENT https://doi.org/10.14324/111.444/ucloe.000057 Spectral bands selection In LULC classification, different land cover classes may respond to different ranges of wavelengths,
and not all spectral bands are useful for the analysis. Consequently, it is imperative to appropriately
identify the useful ranges of wavelength as the procedure increases class discrimination [64]. Chen et al. made an assumption that the low reflectance of mangroves in the short wavelength
infrared (SWIR) region of the electromagnetic spectrum was due to the weak-scattering signal
of the intercellular structure of the leaves [65]. Unsurprisingly, the low reflectance of the mixed
mangrove assemblage with the surrounding mud and water could further reduce the reflected
radiance of mangroves in general. Therefore, they used the Jeffries–Matusita distance technique
to calculate the spectral separability among the LULC classes. This technique was adopted for
this research and was conducted using the spatialEco package version 1.3–7 in R programming
software [66–68]. The Jeffries–Matusita criterion measures the distance between the means of each class feature
and the distribution of values around the means, giving a measure of spectral separability between
the features of the class, and was thus able to determine the quality of the target class samples
[68,69]. Values range from 0 to 2, where 2 indicates high separability while the lower values indicate
a possible misclassification of the classes [70]. In the latter case, distances registered below the
threshold of 1 were removed from the prioritised band image. Additionally, we have considered
the Jeffries–Matusita values between 1.7 and 1.9, as good class separability [63]. In this study we
combined the equivalent bands of each sensor to give an overall distance for the colour band. The
generated results for the Jeffries–Matusita distance calculation indicate that the highest levels of
separability between the mangrove vegetation and non-mangrove vegetation classes were observed
for bands 5–4–3 for TM and ETM+ and 6–5–4 for OLI (Table 1). Thus, the band combination of
SWIR1–NIR–Red was selected as the most appropriate band for the entire image classification. reflectance value for dark objects across the image is 1% and any values greater than zero can
be attributed to the additive effects of haze [41,55,63]. The relatively constant errors removal was
implemented using the formula: [
]
max
min
v
p
min
min
o
s
z
down
(
)
0.01
(
)
255
L
L
T
L
L
DN
E
cos
T
E
θ
π
λ
−
=
+
×
−
×
×
×
+
×
(5) (5) where Lp is the path radiance; DNmin has adopted the histogram technique [60] allowing the haze
DN value to be automatically calculated from the DN frequency histogram of the image; and Tv and Tz
are assumed to be equal in state thereby downward diffusion of radiation at the surface (Edown = 0) is
absent [60]. Figure 2 The DOS assumes that the lowest 5 / 29 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
https://doi.org/10.14324/111.444/ucloe.000057 UCL OPEN ENVIRONMENT Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Image classification and change detection analysis To delineate the mangroves of Palawan, this study used the support vector machine (SVM) classifier
algorithm. This linear supervised non-parametric statistical learning theory has been proven
effective in LULC research [74–76]. The SVM-based classifier requires a training sample and one of
the advantages of this technique is that it can generalise well from a limited amount of training data
compared to alternative methods [74]. This algorithm uses successive executions of a process until
it generates the probabilistic estimates for known and unknown classes. In this entire procedure,
the Bayesian minimum-error decision rule is adopted [77]. The overall accuracy results of SVM depend on the kernel used as well as the chosen kernel’s
parameters and methods [78]. We chose the parameters gamma (G) in the radial basis function
(RBF) kernel and the C hypermeter in SVM to control the error, using the cross-validation (CV)
optimisation technique [79]. We set the default threshold values of 0.091 for G and 100 for penalty
parameter C to gain a lower bias and penalise incorrect classification heavily [75]. The RBF kernel
formula is shown below: (
)
2
,
exp(
|
–
| ),
0
K x x
g x
x
g
=
−
>
′
′
(6 (6) (
)
2
,
exp(
|
–
| ),
0
K x x
g x
x
g
=
−
>
′
′
where ||x − x′||2 is the squared Euclidean distance between two data points, x and x′; g is the user-
defined gamma. Across the series of Landsat data, we created two spectral classes including (a)
mangrove vegetation, that is, intertidal halophytic forests both natural and rehabilitated, and (b)
non-mangrove areas, for example, rivers, estuaries, lakes, sea, tidal mudflats, agricultural areas,
grassland, high- and lowland forests, bushes, residential and industrial areas in rural and urban
regions, aquaculture ponds, salt pans, etc. A random sampling technique was used to select a
minimum of 400 pixels for each spectral class. For all the classified Landsat images, the total
mangrove areas were quantified. Assessing the accuracy of multi-decadal mangrove change is challenging due to the limited
availability of in-situ reference datasets in the time period of interest [80]. In this work, the accuracy
of mangrove classification was assessed using government data derived from the 2010 historical
record of the National Mapping and Resource Information Authority (NAMRIA). Image classification and change detection analysis The training
mangrove forest polygons were validated through the established testing samples and the accuracy
was assessed using the producer’s accuracy, the user’s accuracy, the overall accuracy and the
kappa coefficient values [81]. This study produced >86% overall accuracy results by which the
definite mapping identification of different land use/land cover categories generated valid results
[82]. Furthermore, the kappa analysis for this study generated results >70%. Upon completing the rigorous pre-processing, image classification and validation procedures, we
conducted the change detection for Palawan and the three case study areas of GCRF BC, using
the SCP version 7.9.0 Matera in Quantum in QGIS version 3.22.1 Białowiez˙a, to determine the
magnitude of changes in mangrove vegetation and non-mangrove classes, and the trends of these
changes across three time periods (1988–1998, 1998–2008 and 2008–2020). Cloud patching process, stacking, mosaicking and masking Clouds and cloud shadows have a significant effect on the satellite sensors’ spectral bands
reflectance values [71] and degrade the quality of the sensors’ data [72]. Therefore, the Landsat
database was searched for the clearest satellite images of the study area with the lowest cloud
cover. However, for images where clouds are present, more than one scene from the same epoch Table 1. Spectral separability results using the Jeffries–Matusita distance technique to isolate
the differences between the mangrove vegetation and non-mangrove areas for each band of TM,
ETM+ and OLI sensors Table 1. Spectral separability results using the Jeffries–Matusita distance technique to isolate
the differences between the mangrove vegetation and non-mangrove areas for each band of TM,
ETM+ and OLI sensors
TM bands
ETM+ bands
OLI bands
Band name
Jeffries–Matusita
1
1
2
Blue
0.51
2
2
3
Green
0.75
3
3
4
Red
1.63
4
4
5
NIR
1.86
5
5
6
SWIR 1
1.91
6
6
10
Thermal
0.72
7
7
7
SWIR 2
1.25 6 / 29 UCL OPEN ENVIRONMENT Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines https://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines was acquired to facilitate the cloud patching process using the Fmask algorithm [71,73]. The
selection of different eras was based on the availability of quality data. Thus, the year 2021 was
excluded from the potential list of options because most of the data available were poor in quality. All the selected bands were stacked together and created a seamless mosaic of the study area. The
ocean areas were masked out using the Normalised Difference Vegetation Index with a threshold of
cut-off of 0.5 [65]. Mangrove change projection In the second and third
projection scenarios, we chose the years 2013–2020 datasets to predict the spatial changes of
mangroves for the years 2030 and 2050. Using the IDRISI Environment version 17.00, the Markov
chain transition probability matrix was generated. Mangrove change projection A Markov chain is a stochastic process that describes the likelihood of changing one state to
another [83] through the implementation of neighbourhood rules [84]. The Markovian process has
been implemented in many LULC studies due to its efficiency in future land use prediction [40–
42,85]. In mangrove forest spatial classifications, the integration of the Markov chain model [65] and
its cross-functional application with cellular automata [85,86] is growing considerably. In statistical terms, the Markov chain modelling can effectively make a prediction of the changes
in LULC based on the calculation of the transition probabilities of one system at time t2 with the 7 / 29 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines UCL OPEN ENVIRONMENT https://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines state of the system at time t1 according to the specific year [41,87]. The transition probability matrix
[88] is one of the descriptive tools generated in the process where the mangrove areas transitional
matrix is derived from different mangrove classes [86]. The Markov processes used in this study are
expressed in Eqs (7–9): 2
1
t
t
v
Mv
=
(7) (7) 2
1
t
t
v
Mv
=
where the input LULC proportion column vector corresponds to vt1 and the output vector to vt2; M
is an m × m transition matrix for the time interval Δt = t2 − t1. The development of the probability
transition matrix (pij) can be calculated as follows: 1
q
i
ij
j
n
n
=
=∑
(8)
/
ij
ij
i
p
n
n
=
(9) 1
q
i
ij
j
n
n
=
=∑
/
ij
ij
i
p
n
n
=
(8) (9) /
ij
ij
i
p
n
n
=
/
ij
ij
i
p
n
n
=
where nij is the number of pixels of class i from the first date (current state) that were changed to
class j in the second date (next period); cell ni is in the change detection matrix by row marginal
frequency; q is the total number of classified classes; and pij is the land-cover probabilistic
transition matrix. We have conducted three projections using the Markov chain model. The first
one was the mangrove projection for 2013 using the 1988–1993 datasets. Model validation of the Markovian process We validated the model by comparing the simulated mangrove and non-mangrove areas in 2013
with the observed data in the 2013 ETM+ map. The output was tested with observed values using
the Pearson’s chi-squared (χ2) test to examine the appropriateness of the model: (
)
2
2
O
E
E
χ
−
= ∑
(10) (
)
2
2
O
E
E
χ
−
= ∑ (10) (10) where O represents the simulated value (1988–1993) and E is the actual value of the transition
matrix (2013–2020). The calculated χ2 is compared with the χ2 from the table at alpha-level of 0.05
with (2 – 1)2 degrees of freedom. The land-use change analysis is compatible with the hypothesis of
data independence if the computed χ2 is smaller than the tabled-value χ2. where O represents the simulated value (1988–1993) and E is the actual value of the transition
matrix (2013–2020). The calculated χ2 is compared with the χ2 from the table at alpha-level of 0.05
with (2 – 1)2 degrees of freedom. The land-use change analysis is compatible with the hypothesis of
data independence if the computed χ2 is smaller than the tabled-value χ2. Spatiotemporal distribution of mangroves and comparison with the previous
records Our mapping classification resulted in two major classes, the mangrove forests and non-
mangrove areas. We have presented in Fig. 3 the spatiotemporal distribution of mangroves in
Palawan within the span of 32 years, particularly the time periods of 1988, 1993, 1998, 2003,
2008, 2013, 2018 and 2020. We observed that mangrove forests in Palawan were generally
concentrated around the coastal boundaries, particularly in estuarine fringes, bays, riverbanks
and the margins between land and sea. Based on this study and the previous records, the
mangrove forest cover in Palawan was still relatively high compared with the other provinces in
the Philippines (e.g., [18]). The largest mangrove concentrations in Palawan were found in the eastern part of the island. These
mangroves form dense and continuous stands in Puerto Princesa City (PPC), Bataraza, Balabac
and Brooke’s Point in the south, and in the municipalities of Taytay, Coron, Busuanga, Culion,
El Nido, Aracelli and Dumaran in the north. In PPC, the greatest concentration of mangroves is
generally found in Puerto Princesa Bay, Honda Bay, Ulugan Bay and Turtle Bay. 8 / 29 UCL OPEN ENVIRONMENT Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines https://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philip 9 / 29
Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
UCL OPEN ENVIRONMEN
https://doi.org/10.14324/111.444/ucloe.000057
The classified maps from 1988 to 2020 showed that the largest area of mangroves in Palawan
was recorded in 2020 (60,033.8 ha) while the year 1998 (48,745.3 ha) had the least extent (Fig. 3). The lower total area calculated for 1998 is likely due to misclassification as a result of minor cloud
patches, especially in the northern part of Palawan. Our estimate for this year, however, does not
deviate too far from the estimates in 1993 (50,045.3 ha) and 2003 (52,961.5), respectively. Figure 3
Spatiotemporal distribution of
mangroves in Palawan in a span of
32 years from 1988 to 2020. The classified maps from 1988 to 2020 showed that the largest area of mangroves in Palawan
was recorded in 2020 (60,033.8 ha) while the year 1998 (48,745.3 ha) had the least extent (Fig. 3). The lower total area calculated for 1998 is likely due to misclassification as a result of minor cloud
patches, especially in the northern part of Palawan. Figure 3
Spatiotemporal distribution of
mangroves in Palawan in a span of
32 years from 1988 to 2020. Spatiotemporal distribution of
mangroves in Palawan in a span of
32 years from 1988 to 2020. Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines In consideration of the funder of this study, we also separately quantified the mangrove extents in
PPC, Aborlan and Taytay. Two of the GCRF BC’s smaller geographical case study areas (barangay)
were located in Aborlan municipality while PPC and Taytay municipality both had four case study
locations each. Among these three major boundaries, Taytay had the largest mangroves cover
followed by PPC and Aborlan (Fig. 4). The mangrove areas in Taytay showed an increase since 1988
(3865.1 ha) and peaked in 2008 (7591.8 ha) before the trend showed a gradual decrease until the
most recent estimate in 2020 (7103.6 ha). Similarly, the mangroves in PPC also exhibited a pattern
of increase from 1988 (2876.3 ha) and reached the highest records in 2008 (6621.4 ha) and 2013
(6738.1 ha) before the total estimates dropped. Unlike the two previous locations, the mangrove
forests in the municipality of Aborlan demonstrated an increasing trend from 1993 (1287.7 ha) to
2020 (1839.7 ha). However, the total mangrove area in Aborlan accounts for only about <25% and
<30% of the overall mangrove forest covers in Taytay and PPC, respectively. Figure 4
Composite representation of area
statistics of mangroves in Palawan
(left y-axis), PPC, Taytay and Aborlan
(right y-axis). One of the most challenging aspects of classifying the non-mangrove areas in this study was
the areal immensity of Palawan. The largest estimate for non-mangrove areas was recorded in
1998 at 1,375,197.7 ha (Fig. 5). Mainly, the non-mangrove areas identified were highland and
lowland forests, agricultural areas and built-up areas (e.g., residential and industrial areas in rural
and urban localities). A trend of decrease in non-mangrove areas was evident from 1998 to 2020
(1,363,909.2 ha). The smallest change, at approximately 250 ha, was recorded between 2018
(1,364,168.1 ha) and 2020. To visualise the mangrove forests extents in Palawan across the different time periods, which used
different techniques and resources, the result of this study particularly for the years 2020, 2018 and
2013 were presented along with other previous estimates. As shown in Fig. 6, our estimates for the
total areal extent of mangrove forests in Palawan are similar to other estimates from 1992 to 2015,
except for the estimate of [89] at only 43,000 ha which was the lowest among all the gathered data. Composite representation of area
statistics of mangroves in Palawan
(left y-axis), PPC, Taytay and Aborlan
(right y-axis). Spatiotemporal distribution of mangroves and comparison with the previous
records Our estimate for this year, however, does not
deviate too far from the estimates in 1993 (50,045.3 ha) and 2003 (52,961.5), respectively. Spatiotemporal distribution of
mangroves in Palawan in a span of
32 years from 1988 to 2020. 9 / 29 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines UCL OPEN ENVIRONMENT https://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Representation of mangrove forest
areas in Palawan based on the previous
estimates (grey bars) and the results of
this study (blue bars). all the references cited in this study, NAMRIA recorded the highest estimates at 63,532 ha in 2010
[15] which was higher than the GMW data in the same year (53 731 ha) and even higher than our
Figure 6
51,438
50,602
(NAMRIA-JAFTA 1992)
50,045
51,346 (NAMRIA 1998)
48,745
52,962
58,400 (PCSD 2005)
53,957 (GMW)
53,924 (GMW)
53,877
53,868 (GMW)
63,532 (NAMRIA 2010)
53,731 (GMW)
56,261 (Long & Giri 2011)
55,302
43,000 (Songcuan et al. 2015)
53,397 (GMW)
53,393 (GMW)
59,774
60,034
0.0
5,000.0
10,000.0
15,000.0
20,000.0
25,000.0
30,000.0
35,000.0
40,000.0
45,000.0
50,000.0
55,000.0
60,000.0
65,000.0
70,000.0
75,000.0
80,000.0
Mangrove forest cover (Ha)
Year
* Estimates from this study
1,372,504.8
1,373,897.7
1,375,197.7
1,370,981.5
1,370,065.8
1,368,640.6
1,364,168.1 1,363,909.2
1,358,000
1,360,000
1,362,000
1,364,000
1,366,000
1,368,000
1,370,000
1,372,000
1,374,000
1,376,000
1,378,000
1988
1993
1998
2003
2008
2013
2018
2020
Non-mangrove area cover (Ha)
Year
Figure 5
Estimated total cover of non-mangrove
areas in Palawan from 1988 to 2020. 1,372,504.8
1,373,897.7
1,375,197.7
1,370,981.5
1,370,065.8
1,368,640.6
1,364,168.1 1,363,909.2
1,358,000
1,360,000
1,362,000
1,364,000
1,366,000
1,368,000
1,370,000
1,372,000
1,374,000
1,376,000
1,378,000
1988
1993
1998
2003
2008
2013
2018
2020
Non-mangrove area cover (Ha)
Year Figure 5 Estimated total cover of non-mangrove
areas in Palawan from 1988 to 2020. 51,438
50,602
(NAMRIA-JAFTA 1992)
50,045
51,346 (NAMRIA 1998)
48,745
52,962
58,400 (PCSD 2005)
53,957 (GMW)
53,924 (GMW)
53,877
53,868 (GMW)
63,532 (NAMRIA 2010)
53,731 (GMW)
56,261 (Long & Giri 2011)
55,302
43,000 (Songcuan et al. 2015)
53,397 (GMW)
53,393 (GMW)
59,774
60,034
0.0
5,000.0
10,000.0
15,000.0
20,000.0
25,000.0
30,000.0
35,000.0
40,000.0
45,000.0
50,000.0
55,000.0
60,000.0
65,000.0
70,000.0
75,000.0
80,000.0
Mangrove forest cover (Ha)
Year
* Estimates from this study * Estimates from this study Mangrove forest cover (Ha) Year all the references cited in this study, NAMRIA recorded the highest estimates at 63,532 ha in 2010
[15], which was higher than the GMW data in the same year (53,731 ha) and even higher than our
most recent estimate for 2020. Our current study revealed a minor difference in the increase of
mangrove forests, showing at least 59,774.2 ha in 2018 and 59,9925.8 ha in 2020, respectively
(Fig. 6). Surprisingly, the mangrove forests assessment of Long and Giri [18] revealed a sudden
decrease in mangrove areas in just a year span. Our estimates for 2013 at 55,302.4 ha had a minor
margin of difference with the approximation obtained by Long and Giri [18]. all the references cited in this study, NAMRIA recorded the highest estimates at 63,532 ha in 2010
[15], which was higher than the GMW data in the same year (53,731 ha) and even higher than our
most recent estimate for 2020. Our current study revealed a minor difference in the increase of
mangrove forests, showing at least 59,774.2 ha in 2018 and 59,9925.8 ha in 2020, respectively
(Fig. 6). Surprisingly, the mangrove forests assessment of Long and Giri [18] revealed a sudden
decrease in mangrove areas in just a year span. Our estimates for 2013 at 55,302.4 ha had a minor
margin of difference with the approximation obtained by Long and Giri [18]. In the 1990s, the earliest records of mangrove estimates were obtained by the Japan Forest and
Technology Association (JAFTA) [90] and NAMRIA. Our current estimate for 1993 (50,045.3 ha) was
much lower compared with the previous records of DENR-JAFTA [90] and NAMRIA at 50,602 ha
and 51,346 ha, respectively. However, our estimate for 1998 (48,745.3 ha) had about a 5% margin
with the NAMRIA’s record (51,346 ha). In 2005, the PCSDS utilised the Satellite Pour I’Observation
de la Terre (SPOT) satellite sensor’s images to delineate the extent of mangroves in Palawan and
generated approximately 58,400 ha. Based on the mangrove data extraction made by Richter
et al. [91] from the Global Mangrove Watch (GMW), in accordance with the same mangrove areal
estimates that were originally created by Bunting et al. [92], the GMW figures from 2007 to 2010
had a very slight difference with the 2008 estimate (53,877 ha) for this study. Unsurprisingly, among Figure 4 1988
1993
1998
2003
2008
2013
2018
2020
Palawan
51,438.2
50,045.3
48,745.3
52,961.5
53,877.2
55,302.4
59,774.9
60,033.8
PPC
2876.3
2056.3
5634.2
5922.7
6621.4
6738.1
6709.4
6601.8
Taytay
3865.1
3311.5
6003.4
6120.8
7591.8
7351.5
7285.1
7103.6
Aborlan
1363.4
1287.7
1591.8
1553.4
1697.6
1842.5
1803.2
1839.7
300
1300
2300
3300
4300
5300
6300
7300
100
5100
10,100
15,100
20,100
25,100
30,100
35,100
40,100
45,100
50,100
55,100
60,100
65,100
Mangrove forest cover (Ha)
Mangrove forest cover (Ha) Mangrove forest cover (Ha) UCL OPEN ENVIRONMENT 10 / 29 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Table 2. Comparison of mangrove forest areas in Taytay, Aborlan and PPC based on the previous
estimates and the results of this study
Year (Reference)
Mangrove forest cover (Ha)
PPC
Taytay
Aborlan
1992 [93]
–
–
1494.8
1993
–
–
1287.7
1998
5634.2*
–
1591.8*
2003
5922.7*
–
–
2003 [27]
3201.8
–
–
2007 [91]
5839.8
6727.1
1340.7
2008 [91]
5835.7
6714.2
1341.3
2008
6621.4*
7591.8*
1697.6*
2009 [91]
5816.3
6713.2
1341.3
2010 [94]
4020.0
1578.0
1202.0
2010 [91]
5773.3
6715.5
1341.3
2013 [27]
4577.2
–
–
2013
6738.1*
7351.5*
1842.5*
2014 [93]
–
–
1866.8
2015 [91]
5754.8
6601.0
1337.2
2016 [94]
5668.0
3905.0
1655.0
2016 [91]
5754.8
6601.0
1337.2
2018
6709.4*
7285.1*
1740.3*
2020
6601.8*
7103.6*
1839.7*
The ‘*’ symbol denotes the estimates from this study. The GMW estimates were sourced from Richter et al. [91]
and are based on the measurements by Bunting et al. [92]. Table 2. Comparison of mangrove forest areas in Taytay, Aborlan and PPC based on the previous
estimates and the results of this study The ‘*’ symbol denotes the estimates from this study. The GMW estimates were sourced from Richter et al. [91]
and are based on the measurements by Bunting et al. [92]. In the municipality of Taytay, our estimated result obtained in 2008 has a close margin of difference
from the GMW data. However, unsurprisingly our estimates for 2013 (7351.5 ha) and 2018
(7103.6 ha) differed significantly from the data gathered by Jansen [94] in 2010 (1578 ha) and 2016
(3905 ha; Table 2). A similar interpretation applies to the data by [94] in 2010 and from the GMW
report in the same year where the former generated a very low estimate (1578 ha) against the latter
figure of 6715.5 ha. Pagkalinawan and Ramos [27] estimated the total mangrove forests extent in PPC at 3201.8 ha. It was less than our calculated results for 1998 (5634.2 ha) and 2003 (5922.7 ha), respectively
(Table 2). On separate assessments, Jansen [94], Bunting et al. [92] and Pagkalinawan and Ramos
[27] recorded 4020 ha, 5773.3 ha and 4577.2 ha of mangrove forests in 2010 and 2013. We
obtained a relatively higher estimate in 2013 (6738.1 ha) compared with Pagkalinawan and Ramos
[27] in the same year. We only observed an almost 100 ha difference between the estimates of
Jansen [94] in 2016 and the quantified extent made by Bunting et al. [92] in the same year. However,
between 2016 and 2020, an almost 1000 ha difference was observed between the previous and
current estimates. Figure 6 The result of mangrove forest covers we obtained in 1993 (1287.7 ha) for Aborlan was
comparably lower than the estimation made by [93] in 1992 (1494.8 ha). However, a small gap in
the estimated values was determined between the work of Venturillo [93] in the same period and
this study in 1998 (1591.8 ha; Table 2). Additionally, this study estimated the mangrove forests in
Aborlan in 2008 at about 1676.6 ha which was higher than the GMW data (1341.3 ha). Although
the interval of years was relatively small between 2010 and 2013, the assessment made by
Jansen [94] in 2010 at 1202 ha was distinctly lower than the estimates from GMV [92] and our
result for 2013 (1842.5 ha). Unsurprisingly, from the time periods 2013–2018, the GVM data for
2015 and 2016 [92] are similar, when in fact variations in areal changes were evident between
2013, 2014 and 2016. However, all the assessments reported for Aborlan revealed a similar
pattern where mangrove forest cover increased from inclusive time periods 1992, 1993, 1998,
2010, 2013, 2014 and 2016. 11 / 29 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
https://doi.org/10.14324/111.444/ucloe.000057 UCL OPEN ENVIRONMENT Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
https://doi.org/10.14324/111.444/ucloe.000057 https://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Mangroves change detection We carried out change detection analysis for mangroves in Palawan by comparing multiple years
in discrete intervals (e.g., 10-year gap, 7-year gap). The results of the change detection statistics
within the four time periods (1988–1998, 1998–2008, 2008–2018, 2013–2020) showed that the
mangrove extents in the Palawan dramatically increased for the last 32 years (Fig. 4, Table 3). The
periods with the greatest change in mangrove forest extents in Palawan were recorded in 2008–
2018 and 1998–2008, showing at least 10.95% (5897.7 ha) and 10.53% (5131.9 ha) increase since
the time periods 1998–2018 (Table 3, Fig. 7a,b). However, we also noted the reduction in mangrove
forest cover during the time period 1988–1998 at 5.24% (2692.9 ha) loss. Although this decrease
might imply disturbance in the mangrove ecosystems in the study area, we did not exclude from
our conclusion that this figure could be attributed to the spectral confusion of the different classes
during the classification stage (see Table A3). Concurrently, the mangrove forest cover in PPC showed a sharp increase from 1988 to 1998 at
about 2757.9 ha (95.88%). However, unlike the increasing trend in Palawan in 2013–2020, the
percentage of change at 2.02% (136.3 ha) in the mangrove forest cover in PPC on the same time
period showed a slight decrease. Most of the mangroves in PPC were found in the eastern seaboard
of the study area, forming dense and narrow canopies along the riverbanks, estuarine regions
and margins of the bays, particularly in Honda Bay, Puerto Bay and Turtle Bay. The only notable
concentration of mangroves in the western seaboard of PPC was found in Ulugan Bay (Fig. 7c). Similarly, the municipality of Taytay also established an increase from the time periods
1988–1998 and 1998–2008 with the percentage of increase at about 55.32% (2138.3 ha) and
26.46% (91,588.4 ha), respectively (Table 3). Since 2008, the mangroves in this region suffered a
consecutive loss, particularly with the reducing rates of 4.04% and 3.37% in 2008–2018 and 2013–
2020, respectively. Despite this decrease, the mangrove extent in Taytay remained relatively higher
than PPC and Aborlan (Fig. 3). These mangroves were mostly concentrated in Taytay Bay and
along the Malampaya Sound area. The thick mangrove assemblages within the inner south-eastern
portion of the Malampaya Sound were notable in the classified map. Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines rank (11.80% and 11.56% omission errors). We presumed that the low overall accuracy and kappa
coefficient values generated for 1993 were due to the poor satellite image quality. During this
period, the cloud cover in two of the six scenes [refer to Table A2 in Appendix A: WRS Path 116/
Row 052 (cloud cover = 3, cloud land cover = 13) and WRS Path 118/Row 054 (cloud cover = 8,
cloud land cover = 20)] made marginal spectral confusion between different features. Generally, our
classifications only produced <15% commission and omission errors for both mangrove forest and
non-mangrove area classes (see Table A3). The percentage of reduction or increase in mangrove extents in each region was quantified based on the calcu-
lation used by [65]: (Sj−Si)/Si × 100, where Sj and Si represent the total areas in each categorical class in the ith
and jth time periods. The symbol ‘▲’ denotes the percentage and areal increase in mangrove forests while the
decrease is denoted by the symbol ‘∇’, respectively. Accuracy assessment Using the 2010 LU/LC NAMRIA map as our ground reference data, the mangrove classification
accuracies for years 1988, 1993, 1998, 2003, 2008, 2013, 2018 and 2020 were generated. The
comparative accuracy measurements yielded satisfactory agreements across all the years. The
highest and lowest overall accuracies and kappa coefficient values for the mangrove forest
class were produced in 2020 (92.90% and 0.91) and 1993 (86.66% and 0.73) classification
maps, respectively (see Figure A1). The highest and lowest user’s accuracy in the classification
of mangrove forest features were generated in the years 2003 (95.76%) and 1993 (86.04%). These suggest the commission errors of 4.24% and 13.96%, in which the pixels identified in
the map as mangrove forest class actually represent an incorrect class based on a reference
image. On the other hand, the generated producer’s accuracy quantifies the probability that
a pixel was classified as something other than that class. The year 2013 yielded the highest
producer’s accuracy (6.73% omission error) and the eras of 1998 and 1993 were at the lowest 12 / 29 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
https://doi.org/10.14324/111.444/ucloe.000057 UCL OPEN ENVIRONMENT https://doi.org/10.14324/111.444/ucloe.000057 Mangroves change detection Furthermore, mangroves were
seen forming boundaries along the coastlines of smaller and larger islands in Taytay Bay, especially
in the north-eastern part of the bay (Fig. 7d). In comparison with the mangrove forests in Taytay and PPC, the municipality of Aborlan only
suffered a small loss in mangrove assemblages during 2013–2020 (0.15%, 2.8 ha; Table 3). For
the period of 20 years, the mangrove forest cover in Aborlan increased, although the extent of
expansion was relatively lower than PPC and Taytay. Despite the similarities in the pattern of
changes in Palawan, we did not exclude the possibility that the variations in tidal inundation and
the time of the data acquisition may influence the estimations. Although we did not exclude the Table 3. Changes in mangrove forest distribution in Palawan during (a) 1988–1998, (b) 1998–2008,
(c) 2008–2018 and (d) 2013–2020
Time period
Palawan
PPC
Taytay
Aborlan
Area (Ha)
%
Area (Ha)
%
Area (Ha)
%
Area (Ha)
%
1988–1998
2692.9∇
5.24∇
2757.9▲
95.88▲
2138.3▲
55.32▲
228.4▲
16.75▲
1998–2008
5131.9▲
10.53▲
987.2▲
17.52▲
1588.4▲
26.46▲
105.8▲
6.65▲
2008–2018
5897.7▲
10.95▲
88.0▲
1.33▲
306.7∇
4.04∇
105.6▲
6.22▲
2013–2020
4731.4▲
8.56▲
136.3∇
2.02∇
247.9∇
3.37∇
2.8∇
0.15∇
The percentage of reduction or increase in mangrove extents in each region was quantified based on the calcu-
lation used by [65]: (Sj−Si)/Si × 100, where Sj and Si represent the total areas in each categorical class in the ith
and jth time periods. The symbol ‘▲’ denotes the percentage and areal increase in mangrove forests while the
decrease is denoted by the symbol ‘∇’, respectively. Table 3. Changes in mangrove forest distribution in Palawan during (a) 1988–1998, (b) 1998–2008,
(c) 2008–2018 and (d) 2013–2020
Time period
Palawan
PPC
Taytay
Aborlan
Area (Ha)
%
Area (Ha)
%
Area (Ha)
%
Area (Ha)
%
1988–1998
2692.9∇
5.24∇
2757.9▲
95.88▲
2138.3▲
55.32▲
228.4▲
16.75▲
1998–2008
5131.9▲
10.53▲
987.2▲
17.52▲
1588.4▲
26.46▲
105.8▲
6.65▲
2008–2018
5897.7▲
10.95▲
88.0▲
1.33▲
306.7∇
4.04∇
105.6▲
6.22▲
2013–2020
4731.4▲
8.56▲
136.3∇
2.02∇
247.9∇
3.37∇
2.8∇
0.15∇ Table 3. Changes in mangrove forest distribution in Palawan during (a) 1988–1998, (b) 1998–2008,
(c) 2008–2018 and (d) 2013–2020 The percentage of reduction or increase in mangrove extents in each region was quantified based on the calcu-
lation used by [65]: (Sj−Si)/Si × 100, where Sj and Si represent the total areas in each categorical class in the ith
and jth time periods. Mangroves change detection The symbol ‘▲’ denotes the percentage and areal increase in mangrove forests while the
decrease is denoted by the symbol ‘∇’, respectively. 13 / 29 UCL OPEN ENVIRONMENT spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines ttps://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines p
p
y
g
g possibility that mangroves can also be found in the western seaboard of Aborlan, for this study we
only recorded the mangroves in the eastern seaboard portion. Notably, the small islands of Puntog
and Malunot generally had thick mangrove assemblages (Fig. 7e). possibility that mangroves can also be found in the western seaboard of Aborlan, for this study we
only recorded the mangroves in the eastern seaboard portion. Notably, the small islands of Puntog
and Malunot generally had thick mangrove assemblages (Fig. 7e). Changes in mangrove forests in
Southern Palawan from 1988 to 2020. There was a clear pattern of change in non-mangrove areas in Palawan from 1988 to 2020. An increasing trend was seen from 1988 to 1998 before a spike of decrease happened. The 14 / 29 14 / 29 UCL OPEN ENVIRONMENT https://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines evidence of decreasing trend continued from 2003 to 2020 (Fig. 4). We assumed that these
changes incorporate growth in closed-forest areas and the residential, industrial and agricultural
developments in the region. Moreover, we also presumed that tourism growth and infrastructure
expansion projects (e.g., construction of national roads or highways) play a critical role in the
elaborated expansion of non-mangrove areas in Palawan. Figure 7b Changes in mangrove forests in
Northern Palawan from 1988 to 2020. 15 / 29 15 / 29 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
https://doi.org/10.14324/111.444/ucloe.000057 UCL OPEN ENVIRONMENT Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines 9
Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
UCL OPEN ENVI
https://doi.org/10.14324/111.444/ucloe.000057
Mangrove forests projection and model’s accuracy
The Markov’s transition probability matrix was generated for the two time periods, 1988–19
2013–2020 (see Appendix A). These numbers suggested the probabilities of change in man
forest and non-mangrove area classes in Palawan. The projected areal extent of mangroves
2013 (52,414.5 ha) corresponds slightly with the observed 2013 extent at 51,438.2 ha (Fig. which indicated fewer variations between the two datasets. For this instance, we confirmed
7c
s in mangrove forests in Puerto
City from 1988 to 2020. Mangrove forests projection and model’s accuracy The Markov’s transition probability matrix was generated for the two time periods, 1988–1993 and
2013–2020 (see Appendix A). These numbers suggested the probabilities of change in mangrove
forest and non-mangrove area classes in Palawan. The projected areal extent of mangroves for
2013 (52,414.5 ha) corresponds slightly with the observed 2013 extent at 51,438.2 ha (Fig. 8a),
which indicated fewer variations between the two datasets. For this instance, we confirmed that The Markov’s transition probability matrix was generated for the two time periods, 1988–1993 and
2013–2020 (see Appendix A). These numbers suggested the probabilities of change in mangrove
forest and non-mangrove area classes in Palawan. The projected areal extent of mangroves for
2013 (52,414.5 ha) corresponds slightly with the observed 2013 extent at 51,438.2 ha (Fig. 8a),
which indicated fewer variations between the two datasets. For this instance, we confirmed that Changes in mangrove forests in Puerto
Princesa City from 1988 to 2020. 16 / 29 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
https://doi.org/10.14324/111.444/ucloe.000057 UCL OPEN ENVIRONMENT https://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines the transition matrices between 1988 and 1993 could be effective for predicting the dynamic
change in the mangrove forests and non-mangrove areas in Palawan. We found that the mangrove forests in the region will likely increase by 8.18% (64,946.3 ha)
d 11 56% (66 972 1 h ) i
th
2030
d 2050 (Fi
8 ) C
l
it
j
t d
7d
in mangrove forests in Taytay
8 to 2020. the transition matrices between 1988 and 1993 could be effective for predicting the dynamics of
change in the mangrove forests and non-mangrove areas in Palawan. Changes in mangrove forests in Aborlan
from 1988 to 2020. 51,438.2
50,045.3
55,302.4
52,414.5
25,000
30,000
35,000
40,000
45,000
50,000
55,000
60,000
1988
1993
2013
2013
Mangrove forest cover (Ha)
Year
(a)
Mangrove forests observed
Mangrove forests predicted
1,372,504.8
1,373,897.7
1,368,640.6
1,400,782.5
100,000
250,000
400,000
550,000
700,000
850,000
1,000,000
1,150,000
1,300,000
1,450,000
1,600,000
1988
1993
2013
2013
Non-mangrove area cover (Ha)
Year
(b)
Non-mangrove areas observed
Non-mangrove areas predicted
55,302.4
60,033.8
64,946.3
66,972.1
20,000
25,000
30,000
35,000
40,000
45,000
50,000
55,000
60,000
65,000
70,000
2013
2020
2030
2050
Mangrove forest cover (Ha)
Year
(c)
Mangrove forests observed
Mangrove forests predicted
1,368,640.6
1,363,909.2
1,302,149.6
1,265,498
50,000
250,000
450,000
650,000
850,000
1,050,000
1,250,000
1,450,000
2013
2020
2030
2050
Non-mangrove area cover (Ha)
Year
(d)
Non-mangrove areas observed
Non-mangrove areas predicted
ges in
angrove
ve forests
time
mangrove
ng time
ove forests
using
d) Non-
or 2030 &
013–2020. 1,372,504.8
1,373,897.7
1,368,640.6
1,400,782.5
100,000
250,000
400,000
550,000
700,000
850,000
1,000,000
1,150,000
1,300,000
1,450,000
1,600,000
1988
1993
2013
2013
Non-mangrove area cover (Ha)
Year
(b) (b) Figure 8 Mangrove forests observed
Mangrove forests predicted
55,302.4
60,033.8
64,946.3
66,972.1
20,000
25,000
30,000
35,000
40,000
45,000
50,000
55,000
60,000
65,000
70,000
2013
2020
2030
2050
Mangrove forest cover (Ha)
Year
(c) (d) Year Non-mangrove areas observed
Non-mangrove areas predicted statistical independence for the data was rejected. Therefore, predictive modelling using the Markov
chain can be used for forecasting mangroves in Palawan. statistical independence for the data was rejected. Therefore, predictive modelling using the Markov
chain can be used for forecasting mangroves in Palawan. Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines We found that the mangrove forests in the region will likely increase by 8.18% (64,946.3 ha)
and 11.56% (66,972.1 ha) in the years 2030 and 2050 (Fig. 8c). Conversely, it was projected
that the non-mangrove areas in Palawan were likely to reduce by 4.53% (1,302,149.6 ha) and
7.21% (1,265,498 ha) in 2030 and 2050, respectively (Fig. 8d). There was a slight increase in We found that the mangrove forests in the region will likely increase by 8.18% (64,946.3 ha)
and 11.56% (66,972.1 ha) in the years 2030 and 2050 (Fig. 8c). Conversely, it was projected
that the non-mangrove areas in Palawan were likely to reduce by 4.53% (1,302,149.6 ha) and
7.21% (1,265,498 ha) in 2030 and 2050, respectively (Fig. 8d). There was a slight increase in 17 / 29 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
https://doi.org/10.14324/111.444/ucloe.000057 UCL OPEN ENVIRONMENT Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines mangrove forests in Palawan for the simulated time period 2030 (64,946.3 ha) compared with 2013
(52,414.5 ha) and 2050 (66,972.1 ha; Fig. 8a,c). Changes in mangrove forests in Aborlan
from 1988 to 2020. The result of the accuracy assessment using the time period 1988–1993 and the projected 2013
output was evaluated using a χ2 test, indicating a value of 150.8 which was larger than 3.841 for
the critical level of P = 0.05 with (2 – 1)2 degrees of freedom. This suggests that the hypothesis of 18 / 29 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines UCL OPEN ENVIRONMENT https://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Although this number seems fairly alarming, the local government of PPC asserted that these
initiatives could promote the smooth spatial expansion of the migration of mangroves in the future
because most of the relocated local residents were previously living within the adjacent areas where
mangroves are located [100]. Prior to the declaration of the protected area networks in Palawan, in 1981 and 1991, the mangrove
areas in the province including the adjacent parcels of mangrove forests in the county were
estimated at 74,267 ha [101]. Following the time after the integration of the SEP law in Palawan
in 1992, the mangrove areas changed significantly [17] with at least 50,045 ha remaining areas
in 1993 (Fig. 4). In contrast, a significant decrease of non-mangrove areas, which was notably
recorded from this study from 1998 to 2018 (Fig. 5), coincides with the time periods where massive
deforestation in the southern part of Palawan led to the reduction in the areal size of the forested
areas during 2003–2010 [102]. Explicitly, we have found a significant increase in non-mangrove
areas between 2013 and 2020, which was approximately 3 years after the implementation of
the National Log Ban and the institutionalisation of an Anti-Illegal Logging Task Force in 2011. Interestingly, according to the report of DENR [103], among all the provinces in the Philippines,
Palawan had the largest areal extent of forestland in 2020, totalling about 1,035,926 ha. We had
identified that this study poses limitations against the generated results about the non-mangrove
area class because we only referred to the generalisation of spectral separability. For this instance,
we recommend that future similar studies should also focus on the spatial dynamics of multiple LU/
LC areas. Based on a joint venture initiative by NAMRIA and JAFTA in 1992, an aerial survey was conducted
in Palawan. Among the notably remotely sensed information they obtained was evidence of small-
scale logging activities, particularly in Taytay, and the slash and burn cultivation ‘Kaingin’ in the
central boundary of PPC (e.g., Honday Bay, Ulugan Bay; Fig. 8b) and across the municipalities of
San Vicente and Taytay [90]. PCSDS [96] further reported that a massive extraction of mangrove
raw products for firewood consumption was rampant in Taytay. Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines 1988 in this study, this major project has been almost completed. Therefore, we deemed that this
condition serves as a good baseline of information to envisage the changes in land use patterns in
Palawan. But perhaps, the major framework for all development undertakings in Palawan was the
passage of the Republic Act 7611 known as the SEP for Palawan Act in 1992. Within this law, the
spatial basis for the implementation of its main goal is the Environmentally Critical Areas Network
(ECAN) Zonation Project [96]. The strategic approach of ECAN is composed of three main components: terrestrial, coastal/
marine zones and tribal ancestral lands. The multiple utilisations of every resource within these
components are defined according to different zones, particularly within the multiple/manipulative
zone and buffer zone. The buffer zone is further divided into three distinct zones where the level of
restriction in resources extraction differs. The buffer zone comprises restricted use area (i.e., where
limited non-consumptive activities may be allowed as long as they will not impair the ecological
balance), controlled use area (i.e., activities such as mining, logging, tourism development, research
and other minor resources extraction may be allowed to operate but must be strictly in compliance
with the law) and traditional use area (i.e., located along the edges of intact terrestrial forests where
traditional use has already been established). The intensive utilisation of land use in Palawan is
clearly defined under the multiple/manipulative use zone areas [97,98]. Due to the ECAN zoning
strategy, multiple land-use areas in Palawan have been assessed, marked and delineated based on
their biophysical or natural and anthropogenic attributes to regulate activities, sustain the ecological
integrity and properly manage the carrying capacity [45]. Gilman et al. [30] and Polidoro et al. [99] asserted that the economic growth and the augmentation
of the human population are two major factors that influence the changes in the extent of mangrove
forests and other land use areas. In PPC specifically, where the greatest housing development
projects in Palawan are generally concentrated, the conception of the city’s housing project in
1992 had managed to transform different land use across its boundaries. For example, the multiple
housing projects in Barangay Sicsican, Mangingisda, San Jose, San Manuel, Bahile, Tagburos, Sta. Cruz and Bahile, converted hundreds of hectares of collective land use areas into residential space. Discussion The course of major development in Palawan was started in 1981 with the implementation of the
Palawan Integrated Area Development Project [95]. Following the acquisition of Landsat data for 19 / 29 UCL OPEN ENVIRONMENT https://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Managed Resources Protected Area in 1998 [104], the results obtained from this study (i.e., Fig. 7c),
suggests a the reason for an increasing trend in mangrove forest cover in Taytay. Correspondingly,
an approximately 8.7% increase in old-growth forest coverage in the protected area of Bacuit Bay
has been reported a year after it become fully protected under the law in 1991 [105]. Moreover,
PCSDS [96] reported that two endemic mangrove species in the Philippines namely, Rhizophora
stylosa and Compostenum philippinnensis, were abundant in the northern part of Palawan including
Taytay. For this reason, we assume that the abundance of their presence in this region contributes
to the successful protection and recovery of mangrove forests. Richter et al. [91] recently reported that communities interviewed generally perceived mangrove
condition in Palawan had improved over the last 10 years. They reported that the perception of the
local communities in Taytay, in reference with the mangrove forest ecosystem quality in their area,
suggested no change in condition compared with the findings from this study that showed a decrease
in extent over the past 10 years, although it is apparent that the extent has increased significantly
over the interviewee’s lifetime [91]. Similarly, they reported that the communities in Aborlan and PPC
perceived an improvement in mangroves over the last 10 years [91]. This study indicates that, while
there was a gain in mangrove extent between 2008 and 2013, since 2013 there has been slight
decline in mangrove cover or cover has remained stable in these areas (Fig. 6, Table 3). The discrepancy in these results could be attributed to the reputation of Palawan for having still
relatively high mangrove forest cover in comparison with the other provinces in the Philippines. The positive outlook of the local communities may be influenced by the environmental regulatory
conceptions where they think that the province has strict regulated forest activities as the entire
mangrove forests in the study area are located within the existing protected area networks (i.e.,
IUCN, SEP Law, ECAN Zoning Project). Also, because local communities were actively involved
in yearly ‘mangrove tree planting’ activities across Palawan, for example, the local government
of PPC has already planted around 800,000 mangroves since 2003 [106], they presume that this
type of activity is a good indicator of a successful mangrove management. However, there were
still no local studies that investigate whether the different mangrove rehabilitation programmes in
Palawan are successful or not. It is also likely that, as this study used lower-to-moderate resolution
satellite data, the ability to detect young mangroves that are small and sparce (i.e., saplings) is low
so these areas may not be included in the extent figures. The perceptions of interviewees may also
indicate improvements in mangrove condition and health, rather than simply on extent of mangrove
coverage, which is information harder to attain by remote sensing. On the other hand, we presumed that a large percentage of change in non-mangrove areas in
Palawan could be attributed to the progressive changes of other ground features in the region
(e.g., deforestation, forest regeneration, infrastructure, industrial and residential developments). For example, in PPC alone, a large portion of the non-mangrove area in the outskirt region of
Barangay Sta. Lourdes, which was previously a part of higher elevated grassland/bushland
region, has been converted into a sanitary landfill. Also, we have noted that the projected changes
in the non-mangrove area class might be attributed to the mining activities in the southern
Palawan, particularly in the municipalities of Bataraza, Brooke’s Point, Aborlan and Narra. Another
contributing element, which we assumed could have a large contribution to the changes in non-
mangrove areas in Palawan, was the inception of the Philippine government’s infrastructure-
growth-targeting programme known as ‘Build! Build! Build’, which was started in the last quarter
of 2016. Major highways, roads and bridges have been expanded or re-constructed across the
country, including in Palawan, which led to the conversion of other land use areas. We expected
that this type of development will continue to transform landscape patterns in Palawan until the
end-term of the current government administration. Lastly, an increase in non-mangrove areas for
the years 2030 and 2050 was also expected due to the influence of tourism demand in Palawan. As
the global Covid-19 pandemic starts to shift to an endemic approach, the tourism industry in the
province is now gradually gaining momentum. For example, this situation spurred global interest to
visit/revisit the region’s historical and popular tourism sites, which had been restricted for almost
2 years due to the global outbreak of Covid-19. The largest projection increment in mangrove aerial extents will be recorded in the next 30 years in
2050. These anthropogenic stresses were
assumed to cause changes in the land use/land cover areas in the northern part of the island during
the pre- and post-establishment of a marine reserve within a small portion of the north-western tip
of mainland Palawan (e.g., Bacuit Bay in El Nido municipality) in 1991. However, following the expansion of the protected areas in northern Palawan (i.e., extension
for 1991 – declared Bacuit Bay Marine Reserve) under the establishment of the El Nido-Taytay 20 / 29 UCL OPEN ENVIRONMENT Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
https://doi.org/10.14324/111.444/ucloe.000057 ttps://doi.org/10.14324/111.444/ucloe.000057 Conclusions Our study demonstrates the capability of the Markov chain model in predicting the future
expanse of mangrove forests in Palawan using the multi-date Landsat satellite images from
1988 to 2020. This study found that in all study areas mangrove extent has increased from 1988
levels, although the trajectories since 2008 are more variable. Our analysis has shown the high
likelihood of an increase in areal extent of mangroves in Palawan, from our most recent estimate
in 2020 (60,033.8 ha) up to the years 2030 (64,946.3 ha) and 2050 (66,972.1 ha). However, these
projections should be considered a baseline and must be interpreted with caution, as this work did
not integrate environmental factors that may or had influenced the changes in mangrove forests. For the moment, it would still be sensible to accept that mangrove forests are under constant
threat especially in the context of global climate change. The impact mechanism of sea-level
rise on mangroves continues to increase as the greenhouse gas emissions persist. Furthermore,
other threats such as coastal conversion, water pollution and raw products extraction are not
slowing down and continue to potentially impact the mangrove ecosystems worldwide. Integrating
mangrove forest projection at the regional scale is vitally important to determine specific resiliency
response to climate change impacts. The potential of the Markov chain model to project the potential changes of mangrove forests and
other land use areas conveys its importance in the future, especially in the contexts of landscape
management, ecological sustainability and policy intervention. However, as we did not create
this type of model to directly assess our current policies, we recommend that future research
should integrate the cellular automata–Markov model as it provides land cover data needed at
different time steps (i.e., pre- and post-policy intervention) (e.g., [42]). This way, research bodies
can evaluate the impacts of different policies (e.g., 1992 SEP Law, 1981 Mangrove Swamp Forest
Reserve) in the future state of mangroves in Palawan. Furthermore, it would be good to conduct a
similar study but it should also focus on the assessment of different LU/LC patterns to determine
whether the demand of development that spurs the decrease or increase of certain features
of non-mangrove areas is beneficial to the environment or not. Funding This study received funding from the Global Challenges Research Fund, Blue Communities, under the United Kingdom
Research and Innovation, with grant agreement NE/P021107/1. Conclusions This approach might alleviate
uncertainties about the state of other multiple land-use areas in Palawan, other than mangrove
forest, and the potential changes can be dissected and utilised for more effective management
applications. It would also be necessary to investigate the pressures of different socio-economic activities of
village communities on the extent of mangrove forests within the different multiple zones (i.e.,
based on the ECAN Zoning Project) as changes in the distribution and intensity of these activities
in response to social and economic drivers have the potential to contribute to changes in LU/LC
areas. Given all the other driving factors that could influence the changes of mangrove forest cover
in Palawan, we further encourage the implementation of spatio-statistical modelling techniques in
the future, where the changes in land-use areas are to be fitted with environmental covariates. We
think that this type of approach is timely, relevant, cost-effective and could enable the evaluation of
different management interventions and policies not only in Palawan but also in the Philippines and
neighbouring Southeast Asian countries. Acknowledgements The authors express gratitude to the Global Challenges Research Fund, Blue Communities, under the United Kingdom Research
and Innovation for the financial support. We also thank the National Mapping and Resource Information Authority for their
provision of the ground reference data. We expected this evaluation following the assumption where the current ‘Build! Build! Build!’
programme of the Philippine government could catch up with rapid urbanisation and population
growth, which could potentially facilitate the optimisation of mangrove forests protection in the
province. This is because we assumed that relocating the local residents living within the coastal
areas could lessen the threat to the mangrove ecosystem and foster community growth. 21 / 29 UCL OPEN ENVIRONMENT spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines ttps://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines References and mangrove habitats in the Philippines. PLoS One. 2013;8(8):e65735. [1]
Brown C, Corcoran E, Herkenrath P, Thonell J. Marine
and Coastal Ecosystems and Human Well-Being:
Synthesis [online]. Nairobi, Kenya: United Nations
Environment Programme, Division of Early Warning
Assessment; 2006 [Accessed 6 December 2021]. Available from: https://wedocs.unep.org/bitstream/
handle/20.500.11822/9461/Marine%20and%20
Coastal%0Ecosystems%20and%20Human%20
WellBeing_%20A%20synthesis%20report%20
based20on%20the%20findings%20of%20the%20
Millennium%20Ecosystems%20Assessment200652. pdf?sequence=3&%3BisAllowed=. [10] Camacho LD, Gevaña DT, Carandang AP, Camacho SC,
Combalicer EA, Rebugio LL, et al. Tree biomass and
carbon stock of a community-managed mangrove forest
in Bohol, Philippines. Forest Sci Technol. 2011;7(4):161–7. [11] Sarhan M, Tawfik R. The economic valuation of
mangrove forest ecosystem services: implications for
protected area conservation. The George Wright Forum. 2018;35(3):341–9. [12] Dixon TG, Pulhin JM, Tapia MA. Chapter 13 – Fostering
climate change mitigation through a community-based
approach: carbon stock potential of community-
managed mangroves in the Philippines. Coastal
Management. Global Challenges and Innovation. Philippines; 2019. p. 271–82. [2]
Mukherjee N, Sutherland WJ, Khan MNI, Berger
U, Schmitz N, Dahdouh-Guebas F, et al. Using
expert knowledge and modeling to define mangrove
composition, functioning, and threats and estimate time
frame for recovery. Ecol Evol. 2014;4:2247–62. [13] Kandasamy K, Bingham B. Biology of mangroves and
mangrove ecosystems. Adv Mar Biol. 2001;40:81–251. [3]
Ball MC, Pidsley SM. Growth responses to salinity
in relation to distribution of two mangrove species,
Sonneratia alba and S. lanceolata, in northern Australia. Funct Ecol. 1995;9(1):77–85. [14] Primavera JH, Sadaba RB, Lebata MJHL, Altamirano
JP. Handbook of Mangroves in the Philippines – Panay. Tigbauan, Iloilo, Philippines [online]. Philippines:
Southeast Asian Fisheries Development Center; 2004. [4]
Liang S, Zhou R, Dong S, Shi S. Adaptation to salinity
in mangroves: implication on the evolution of salt-
tolerance. Chi Sci Bull. 2008;53(11):1708–15. [15] Food and Agriculture Organization of the United Nations
(FAO). Global Forest Resources Assessment Report
2015: Country Report Philippines [online]. Rome: FAO;
2022 [Accessed 20 January 2022]. Available from:
https://www.fao.org/3/az306e/az306e.pdf. [5]
Mendoza A, Alura D. Mangrove structure on the eastern
coast of Samar Island, Philippines. In: Stott DE, Mohtar
RH, Steinhard GC, editors. Sustaining the global
farm. Selected Papers from the 10th International Soil
Conservation Organization Meeting held May 24–29,
1999 at Purdue University and the USDA-ARS National
Soil Erosion Research Laboratory; 2001. [16] Myers N, Mittermeier RA, Mittermeier CG, Fonseca
G, Kent JM. Biodiversity hotspots for conservation
priorities. Nature. 2000;403:853–8. [6]
Giri C, Ochieng E, Tieszen LL, Zhu Z, Singh A, Loveland
T, et al. Declarations and conflicts of interest
Research ethics statement Not applicable to this article. The authors declare no conflicts of interest with this work. The authors declare no conflicts of interest with this work. Authorship contribution CBC conceptualised the study, collected and analysed the data, interpreted the results and wrote the manuscript; ES edited
the manuscript and contributed to the results and discussion sections; PIM revised the manuscript, analysed the data and
suggested to the improvement of data visualisation and in depth interpretation of the results; DC edited the manuscript;
LAC edited the manuscript, supervised the acquisition of reference data, helped during the conceptualisation of the study and
supervised the funding acquisition. All authors have read and approved the final manuscript. 22 / 29 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines UCL OPEN ENVIRONMENT https://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Consent for publication statement The authors declare that research participants’ informed consent to publication of findings – including photos, videos and any
personal or identifiable information – was secured prior to publication. Open data and materials availability statement The datasets generated during and/or analysed during the current study are available from the corresponding author on
reasonable request. Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines around Manila Bay using Landsat satellite imagery and
Mangrove Vegetation Index (MVI). ISPRS Archives. 2021;XLVI-4/W6-2021:103–8. [21] Dodd RS, Ong JE. Future of mangrove ecosystems to
2025. In: Polunin NV, editor. Aquatic ecosystems: Trends
and global prospects. New York: Cambridge University
Press; 2008. p. 172–287. [37] Otsu N. A threshold selection method from gray-level
histograms. IEEE Trans Systs Man Cybern. 1979;9(1):
62–6. [22] Department of Environment and Natural Resources
(DENR). Sustaining our Coasts: The Ridge-to-Reef
Approach – A Compilation of Technical and Policy
Papers: Mangrove Management. Integrated Coastal
Resources Management Project (ICRMP) of the
Department of Environment and Natural Resources
[online]. Philippines: DENR; 2013 [Accessed 15
December 2021]. Available from: https://faspselib. denr.gov.ph/sites/default/files//Publication%20Files/2. MANGROVES_FINAL.pdf. [38] Gagniuc PA. Markov Chains: From Theory to
Implementation and Experimentation [online]. 2017 [Accessed 18 December 2021]. https://doi. org/10.1002/9781119387596. [39] Yang X. Markov Chain and its applications. MA thesis. Greencastle, IN: DePauw University; 2020 [Accessed
18 December 2021]. https://doi.org/10.13140/
RG.2.2.12289.61287. [23] Lucas RM, Ellison JC, Mitchell A, Donnelly B, Finlayson
M, Milne AK. Use of stereo aerial photography for
quantifying changes in the extent and height of
mangroves in tropical Australia. Wetl Ecol Manag. 2002;10(2):1510175. [40] Coppedge BR, Engle DM, Fuhlendorf SD. Markov
models of land cover dynamics in a southern Great
Plains grassland region. Landsc Ecol. 2007;22:
1383–93. [24] Komiyama A, Ong JE, Poungparn S. Allometry, biomass,
and productivity of mangrove forests: a review. Aqua
Bot. 2008;89:128–37. [41] Kanjirappuzha R, Mukherjee CK, Vinu Chandran R,
Prakash Mohan MM. Land-cover change dynamics
and coastal aquaculture development: a case study
in the East Godavari delta, Andhra Pradesh, India
using multi-temporal satellite data. Int J Remote Sens. 2010;31(16):4423–42. [25] Suratman M. Remote sensing technology: recent
advancements for mangrove ecosystems. In: Faridah-
Hanum I, Latiff A, Hakeem K, Ozturk M, editors. Mangrove ecosystems of Asia. New York: Springer; 2014. [42] Adhikari S, Southworth J. Simulating forest cover
changes of Bannerghatta National Park based on a
CA-Markov model: a remote sensing approach. Remote
Sens. 2012;4:3215–43. [26] Long J, Napton D, Giri C, Graesser J. A mapping and
monitoring assessment of the Philippines’ mangrove
forest from 1990 to 2010. J Coast Res. 2014;30(2):260–71. [43] Adegbola PA, Adewumi JR, Obiora-Okeke OA. Application of Markov chain model and ArcGIS in land
use projection of Ala river catchment, Akure, Nigeria. Niger J Technol Dev. 2021;18(1);1–9. [27] Pagkalinawan HM, Ramos RV. Change detection of
mangrove forest cover in the city of Puerto Princesa
Palawan between 2003 and 2013 using Landsat
imagery [online]. 2013 [Accessed 14 January 2022]. Available from: https://www.academia.edu/9733267/
CHANGE_DETECTION_OF_MANGROVE_FOREST_
COVER_IN_THE_CITY_OF_PUERTO_PRINCESA_
PALAWAN_BETWEEN_2003_AND_2013_USING_
LANDSAT_IMAGERY. [44] Worthington T, Andradi-Brown D, Bhargava R, Buelow
C, Bunting P, Duncan C, et al. Harnessing big data to
support the conservation and rehabilitation of mangrove
forests globally. One Earth. 2020;2:385–486. [45] Palawan Council for Sustainable Development (PCSD). State of the Environment 2009 Updates, Province of
Palawan, Philippines [online]. Puerto Princesa City:
PCSD; 2010 [Accessed 15 January 2022]. Available
from: https://www.pkp.pcsd.gov.ph/SOE/red%20
2010%20_%20State%20of%20Environment%20
Updates%202009,%20Palawan.pdf. [28] Koh HL, Teh SY. Climate change mitigation and
adaptation: role of mangroves in Southeast Asia. In: Leal
Filho W, Azul AM, Brandli L, Özuyar PG, Wall T, editors. Climate action. Encyclopedia of the UN sustainable
development goals. Cham: Springer; 2020. [29] Food and Agriculture Organizations of the United
Nations (FAO). Strategic Environmental Plan for Palawan
Act (RA 7611) [online]. Rome: FAO; 2021 [Accessed 17
December 2021]. Available from: https://www.fao.org/
faolex/results/details/en/c/LEX-FAOC019797. [46] Carandang AP, Camacho LD, Gevaña DT, Dizon JT,
Camacho SC, de Luna CC, et al. Economic valuation
for sustainable mangrove ecosystems management in
Bohol and Palawan, Philippines. Forest Sci Technol. 2013;9(3):118–25. [30] Gilman EL, Ellison J, Duke NC, Field C. Threats to
mangroves from climate change and adaptation
options: a review. Aquat Bot. 2008;89(2):237–50. [47] Philippine Statistics Authority (PSA). Palawan QuickStat
as of April 2009. Compiled by the Databank and
Information Services Division [online]. Philippines: PSA;
2009 [Accessed 15 January 2022]. Available from:
https://psa.gov.ph/sites/default/files/attachments/ird/
quickstat/April_29.pdf. [31] Lillesand TM, Kiefer RW, Chipman JW. Remote sensing
and image interpretation. 7th ed. New York: John Wiley &
Sons Ltd.; 2015. [32] Bolstad P, Lillesand TM. Rapid maximum likelihood
classification. Photogramm Eng Remote Sens. 1991;57:67–74. [48] Sandalo RM, Baltazar T. The Palawan biosphere
reserve. In: Working Papers, South–South Cooperation
Programme on Environmentally Sound Socio-
economic Development in the Humid Tropics. No. 19. UNESCO; 1997. pp. 1–32 [Accessed 2 January 2022]. Available from: https://www.yumpu.com/en/document/
read/6017664/the-palawan-biosphere-reserve-
philippines-unesdoc-unesco. [33] Benediktsson JA, Swain PH, Ersoy OK. Neural network
approaches vs. statistical methods in classification of
multisource remote sensing data. IEEE Trans Geosci
Remote Sens. 1990;28(4):489–92. [34] Ma L, Liu Y, Zhang X, Ye Y, Yin G, Johnson B. Deep
learning in remote sensing applications: a meta-analysis
and review. ISPRS J Photogramm Remote Sens. 2019;152:166–77. [49] Naz AC. References Status and distribution of mangrove forests of
the world using earth observation satellite data. Glob
Ecol Biogeogr. 2011;20:154–9. [17] Palawan Council for Sustainable Development Staff
(PCSDS). State of the Environment 2015 Updates,
Province of Palawan, Philippines [online]. Puerto
Princesa City: PCSD; 2015 [Accessed 1 February
2022]. Available from: https://pcsd.gov.ph/wp-content/
uploads/2020/12/PCSDS-2015-Annual-Report.pdf. [7]
Nagelkerken I, Roberts CM, van der Velde G,
Dorenbosch M, van Riel MC, Cocheret de la Moriniere E,
et al. How important are mangroves and seagrass beds
for coral-reef fish? The nursery hypothesis tested on an
island scale. Mar Ecol Prog Ser. 2002;244:299–305. [18] Long J, Giri C. Mapping the Philippines’ mangrove forests
using Landsat imagery. Sensors. 2011;11:2972–81. [19] Thomas N, Lucas R, Bunting P, Hardy A, Rosenqvist
A, Simard M. Distribution and drivers of global
mangrove forest change, 1996–2010. PLos One. 2017;12:e0179302. [8]
Nagelkerken I, Blaber S, Bouillon S, Green P, Haywood
M, Kirton LG, et al. The habitat function of mangroves
for terrestrial and marina fauna: a review. Aqua Bot. 2008;89(2):155–85. [20] Primavera JH. Development and conservation of the
Philippine mangroves: institutional issues. Ecol Econ. 2000;35(1):91–106. [9]
Honda K, Nakamura Y, Nakaoka M, Uy WH, Fortes
MD. Habitat use by fishes in coral reefs, seagrass beds 23 / 29 UCL OPEN ENVIRONMENT Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines https://doi.org/10.14324/111.444/ucloe.000057 [76] Talukdar S, Singha P, Mahato S, Shahfahad Pal S, Liou
Y-A, Rahman A. Land-use land-cover classification by
machine learning classifiers for satellite observations – a
review. Remote Sens. 2020;12(7):1135. [58] Schroeder TA, Cohen WB, Song C, Canty MJ, Yang Z. Radiometric correction of multi-temporal Landsat data
for characterization of early successional forest patterns
in western Oregon. Remote Sens Envir. 2006;103:
16–26. [77] Press WH, Teukolsky SA, Vetterling WT, Flannery BP,
editors. Numerical recipes: The art of scientific
computing. 3rd ed. Cambridge: Cambridge University
Press; 2007. [59] United States Geological Survey (USGS). Landsat 7 (L7)
Data Users Handbook. LSDS-1927 Version 2.0 [online]. Sioux Falls, South Dakota: EROS; 2019 [Accessed 16
December 2021]. Available from: https://prd-wret.s3.us-
west-2.amazonaws.com/assets/palladium/production/
atoms/files/LSDS1927_L7_Data_Users_Handbook-v2. pdf. [78] Huang C, Davis LS, Townshed JRG. An assessment of
support vector machines for land cover classification. Int J Remote Sens. 2002;23(4):725–49. [79] Buitre MJC, Zhang H, Lin H. The mangrove forests
change and impacts from tropical cyclones in the
Philippines using time series satellite imagery. Remote
Sens. 2019;11:688. [60] Chavez PS, Jr. An improved dark-object subtraction
technique for atmospheric scattering correction of
multispectral data. Remote Sens Environ. 1998;24:
459–79. [80] Liu M, Zhang H, Lin G, Lin H, Tang D. Zonation and
directional dynamics of mangrove forests derived from
time-series satellite imagery in Mai Po, Hong Kong. Sust. 2008;10(6):1913. [61] Song C, Woodcock CE, Seto KC, Lenney MP, Scott
AM. Classification and change detection using landsat
TM data: when and how to correct atmospheric
effects? Remote Sens Environ. 2001;75(2):230–44. [81] Congalton RG. A review of assessing the accuracy of
classifications of remotely sensed data. Remote Sens
Environ. 1991;37(1):35–46. [62] Richards J, editor. Remote sensing digital image analysis:
An introduction. 5th ed. New York: Springer; 2013. [82] Weng QH, editor. Remote sensing and GIS integration:
Theories, Methods, and Applications. New York:
McGraw-Hill; 2010. [63] Jensen JR, editor. Introductory digital image processing –
A remote sensing perspective. 2nd ed. Englewood Cliffs,
Upper Saddle River, NJ: Prentice Hall; 1996. [83] Serfozo R. Markov Chain. In Basics of Applied
stochastic processes. Probability and its application. Springer Science & Business Media, 2009:1–443. [64] Mausel PW, Kramber WJ, Lee JK. Optimum band
selection for supervised classification of multispectral
data. Photogramm Eng Remote Sens. 1990;56(1):55–60. [84] Aitkenhead MJ, Aalders IH. Predicting land cover using
GIS, Bayesian and evolutionary algorithm methods. J
Environ Manage. 2009;90(1):236–50. Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines [67] Kailath T. The Divergence and Bhattacharyya measures
in signal selection. IEEE Trans Commun. 1967;15:52–60. region-by-region overview. Ecosyst Health Sust. 2016;2(4):301211. region-by-region overview. Ecosyst Health Sust. 2016;2(4):301211. [68] Bruzzone LF, Roli SB, Serpico. An extension to
multiclass cases of the Jeffries–Matusita distance. IEEE
Trans Geosci Remote Sens. 1995;33(6):1318–21. [51] Climate Change Commission (CCC). National Integrated
Climate Change Database and Information Exchange
System [online]. Philippines: CCC; 2021 [Accessed 25
February 2022]. Available from: https://niccdies.climate. gov.ph/files/documents/National%20Framework%20
Strategy%20on%20Climate%20Change%20-NFSCC-. pdf. [69] Ghoggali N, Melgani F. Automatic ground-truth
validation with genetic algorithms for multispectral
image classification. IEEE Trans Geosci Remote Sens. 2009;47(7):2172–81. [52] The Climate Reality Project. How is Climate Change
Affecting the Philippines? Philippines: The Climate
Project; 2016 [Accessed 20 January 2022]. Available
from: https://www.climaterealityproject.org/blog/how-
climate-change-affecting-philippines. [70] Richards JA, Jia X, editors. Remote sensing digital image
analysis. Berlin Heidelberg: Springer-Verlag; 1999. [71] Zhu Z, Woodcock CE. Object-based cloud and cloud
shadow detection in Landsat imagery. Remote Sens
Environ. 2012;118(15):83–94. [53] Gordon HR. Calibration requirements and methodology
for remote sensors viewing the ocean in the visible. Remote Sens Environ. 1987;22(1):103–26. [72] Li J, Roy D. A global analysis of sentinel-2A, sentinel-2B
and landsat-8 data revisit intervals and implications for
terrestrial monitoring. Remote Sens. 2017;9(9):902. [54] Lillesand TM, Kiefer RW. Remote sensing and image
interpretation. 3rd ed. New York: John Wiley and Sons
Ltd.; 1994. [73] Zhu Z, Wang S, Woodcock CE. Improvement and
expansion of the Fmask algorithm: cloud, cloud shadow,
and snow detection for Landsat 2015 check title and
Sentinel images. Remote Sens Environ. 2015;159:269–77. [55] Bruce CM, Hilbert DW. Pre-processing Methodology
for Application to Landsat TM/ETM+ Imagery of the
Wet Tropics [online]. Cairns, Australia: Rainforest CRC;
2006 [Accessed 20 December 2021]. Available from:
https://rainforest-crc.jcu.edu.au/publications/landsat_
preprocessing.pdf. [74] Mountrakis G, Im J, Ogole C. Support vector machines
in remote sensing: a review. ISPRS J Photogramm
Remote Sens. 2010;66(3):247–59. [75] Campomanes F, Pada AV, Silapan J. Mangrove
classification using support vector machines and random
forest algorithm: a comparative study. In: GEOBIA
2016: Solutions and Synergies, 14–16 September 2016,
University of Twente Faculty of Geo-Information and
Earth Observation. Enschede: Netherlands; 2016. [56] Young NE, Anderson RS, Chignell SM, Vorster AG,
Lawrence R, Evangelista PH. A survival guide to
Landsat preprocessing. Ecology. 2017;98(4):920–32. [57] Dai X. The effects of image misregistration on the
accuracy of remotely sensed change detection. IEEE
Trans Geosci Remote Sens. 1998;36(5):1566–77. Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines The State of the Philippine Environment: an
Update on Chapter 4 of the 1994 Philippine Human
Development Report. HDN Discussion Paper Series
2012/2013, No. 10 [online]. Philippines: Human
Development Network; 2013 [Accessed 15 January
2022]. Available from: https://www.hdn.org.ph/wp-
content/uploads/DP_10_Naz.pdf. [35] Almahasheer H, Aljowair A, Duarte CM, Irigoien X. Decadal stability of Red Sea mangroves. Estuar Coast
Shelf Sci. 2016;169:164–72. [36] Conopio M, Baloloy AB, Medina J, Blanck AC. Spatio-
temporal mapping and analysis of mangrove extents [50] Ward RD, Friess DA, Day RH, MacKenzie RA. Impacts
of climate change on mangrove ecosystems: a 24 / 29 UCL OPEN ENVIRONMENT Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines https://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines [100] Puerto Princesa City Government. Comprehensive
Housing Program [online]. Puerto Princesa City,
Philippines: PPCG; 2012 [Accessed 8 January
2022]. Available from: https://puertoprincesa. ph/?q=government/city-mayors-initiatives/
comprehensive-housing-program. assemblages and future prediction of the Bangladesh
Sundarbans using Markov chain model and cellular
automata. Environ Sci Process Impacts. 2015;17:1111. assemblages and future prediction of the Bangladesh
Sundarbans using Markov chain model and cellular
automata. Environ Sci Process Impacts. 2015;17:1111. [87] Kumar S, Radhakrishnan N, Mathew S. Land use
change modelling using a Markov model and remote
sensing. Geomat Nat Haz Risk. 2014;5(2):145–56. [101] Melana DM, Melana EE, Mapalo AM. Mangrove
Management and Development in the Philippines –
Report of the Regional Technical Consultation for the
Development of Code of Practice for Responsible
Aquaculture in Mangrove Ecosystems [online]. Philippines: Southeast Asian Fisheries Development
Center; 2000 [Accessed 20 January 2022]. Available
from: https://repository.seafdec.org.ph/bitstream/
handle/10862/712/RTCCode_p39-47.pdf. [88] Omar NQ. Modelling land-use and land-cover
changes using Markov-CA, and multiple decision
making in Kirkuk City. Int J Sci Res Environ Sci. 2014;2(1):29–42. [89] Songcuan AJG, Baloloy AB, Blanco AC, David LT,
Go GA, Cadalzo IE, et al. Mangrove Forest Extent
Mapping in Southwestern Luzon Using Landsay
Imagery. Technical Presentations [online]. 2015
[Accessed 10 December 2021]. Available from: https://
mangroveecology.files.wordpress.com/2017/04/3-2-
mangrove-forest-extent-mapping-in-southwestern-
luzon-using-2015-landsat-imagery.pdf. [102] Wealth Accounting and the Valuation of Ecosystem
Services (WAVES). Pilot Ecosystem Account for Southern
Palawan. Final report of the Technical Working Group
for Southern Palawan ecosystem accounting [online]. Philippines: WAVES; 2016 [Accessed 29 February 2022]. Available from: https://www.wavespartnership.org/sites/
waves/files/kc/WB_Southern%20Palawan%20Tech%20
Report_FINAL_Nov%202016.pdf. [90] Department of Environment and Natural Resources-
National Mapping and Resource Authority, and Japan
Forest Technology Association (DENR-NAMRIA and
JAFTA). Land Cover Statistics 1993–2000: Palawan
Forest Cover Map [online]. Philippines: DENR-JAFTA;
1992–2000 [Accessed 16 December 2021]; Available
from: https://openjicareport.jica.go.jp/pdf/11353935_02. pdf. [103] Department of Environmental and Natural Resources
(DENR). 2011 Philippine Forestry Statistics [online]. DENR, Philippines: Forest Management Bureau; 2011
[Accessed 14 December 2021]. Available from: https://
forestry.denr.gov.ph/images/contents/pdfs/PFS2011.pdf. [91] Richter I, Roberts BR, Sailley SF, Sullivan E, Cheung
VV, Eales J, et al. Building bridges between natural and
social science disciplines: a standardized methodology
to combine data on ecosystem quality trends. Phil Trans
R Soc. 2022;377:20210487. [104] National Integrated Protected Areas Programme
(NIPAP). A Special Project of the Department of the
Department of Environment and Natural Resources
supported with a grant from the European Union
(B7-504 I/93/20), Final Report [online]. Philippines:
NIPAP; 2001 [Accessed 4 March 2022]. [110] Kumar S, Radhakrishnan N, Mathew S. Land use
change modelling using a Markov model and remote
sensing. Geomat Nat Haz Risk. 2014;5(2):145–56. [98]
Aguilla CPA, Gapay IG, Nonato QIC, Simpao ADP,
Tirol RPC. Municipality of El Nido ECAN Resource
Management Plan 2015–2020 [online]. 2020 [Accessed
5 January 2022]. Available from: https://pcsd.gov.ph/
wp-content/uploads/2020/12/3-Municipality-of-El-
NidoECAN-Resource-Management-Plan-2015-2020.pdf. Available
from: https://faspselib.denr.gov.ph/sites/default/files//
Publication%20Files/Technical%20Report_0.pdf. [92] Bunting P, Rosenqvist A, Lucas RM, Revelo L-M,
Hilarides L, Thomas N, et al. The global mangrove
watch-a new 2010 global baseline of mangrove extent. Remote Sens [online]. 2018;10(10):1669. [105] Japan International Cooperation Agency (JICA). The Study on Environmentally Sustainable Tourism
Development Plan for Northern Palawan in the
Republic of the Philippines. Supplemental Report
No. 2, Terrestrial Environment of Northern Palawan
[online]. Philippines; 1997 [Accessed 10 January
2021]. Available from: https://openjicareport.jica.go.jp/
pdf/11374626_01.pdf. [93] Venturillo R. Spatio-temporal mapping, biomass, and
carbon stock assessment of mangrove forest in Aborlan,
Palawan, Philippines. JNS. 2016;15(2):90–103. [94] Jansen L. A remote-sensing based assessment
of ecosystem services for Palawan, The
Philippines. Netherlands: MA thesis, Wageningen
University; 2017. [95] Baylon MCO, Franco RCC, Perez MAB. An economic
analysis of the integrated area development approach
as an alternative to rural development: a case study of
the Palawan integrated area development project phase
1 (1982–90). DLSU (B&E). 1993;5(1). [106] City Tourism Department of Puerto Princesa. Love Affair
with Nature [online]. Puerto Princesa City, Philippines:
CTDPP; 2015 [Accessed 18 December 2021]. Available
from: https://tourism.puertoprincesa.ph/?q=events/love-
affair-nature. [96] Palawan Council for Sustainable Development Staff
(PCSDS). Palawan Council for Sustainable Development
Staff Accomplishment Report CY 2005 [online]. Puerto
Princesa City: PCSD; 2005 [Accessed 15 January
2022]. Available from: https://pcsd.gov.ph/pcsd-2005-
accomplishment-report/. [107] Congalton RG. A review of assessing the accuracy of
classifications of remotely sensed data. Remote Sens
Environ. 1991;37(1):35–46. [108] Kanjirappuzha R, Mukherjee CK, Vinu Chandran R,
Prakash Mohan MM. Land-cover change dynamics
and coastal aquaculture development: a case study
in the East Godavari delta, Andhra Pradesh, India
using multi-temporal satellite data. Int J Remote Sens. 2010;31(16):4423–42. [97]
Palawan Council for Sustainable Development Staff
(PCSDS). Mainstreaming the Environmentally Critical
Areas Network (ECAN) into the Local Land Use Planning
System of the Local Government Units (LGU): Framework
and Methods [online]. Puerto Princesa City, Phillipines:
PCSD; 2015 [Accessed 15 January 2022]. Available from:
https://pkp.pcsd.gov.ph/images/Mainstreaming%20
ECAN%20into%20Comprehensive%20Land%20
and%20Water%20Use%20Plan.pdf. [109] Murugesan S, Zhang J, Vittal V. Finite state Markov
chain model for wind generation forecast: a data-driven
spatiotemporal approach. IEEE PES Innovative Smart
Grid Technologies (ISGT); 2012. https://doi.org/10.1109/
ISGT.2012.6175764. [65] Chen C-F, Son N-T, Chang N-B, Chen C-R, Chang L-Y,
Valdez M, et al. Multi-decadal mangrove forest change
detection and prediction in Honduras, Central America,
with Landsat imagery and a Markov chain model. Remote Sens. 2013;5(12):6408–26. [85] Abdulrahman AI, Ameen SA. Predicting Land use and
land cover spatiotemporal changes utilizing CA-Markov
model in Duhok district between 1999 and 2033. Acad J
Nawroz U. 2020;9(4):71–80. [66] Bhattacharyya A. On a measure of divergence
between two statistical populations defined by their
probability distributions. New Bull Calcutta Math Soc. 1943;35:99–109. [86] Mukhopadhyay A, Mondal P, Barik J, Chowdhury
SM, Ghosh T, Hazra S. Changes in mangrove species 25 / 29 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines UCL OPEN ENVIRONMENT https://doi.org/10.14324/111.444/ucloe.000057 Accuracy assessment Using the 2010 LU/LC NAMRIA map as our ground reference data, the mangrove classification
accuracies for years 1988, 1993, 1998, 2003, 2008, 2013, 2018 and 2020 were generated (Fig. A1). The training mangrove forest polygons were validated through the established testing samples
and the accuracy was assessed using the producer’s accuracy, the user’s accuracy, the overall
accuracy and the kappa coefficient values [107]. Sourced dataset The TM, ETM+ and OLI datasets in multiple years 1988, 1993, 1998, 2003, 2008, 2013, 2018 and
2020 were sourced using the Semi-Automatic Classification Plugin (SCP) version 7.9.0 Matera in
Quantum Geographical Information System (QGIS) version 3.22.1 Białowiez˙a (Table A2). Landsat sensors used The multi-temporal resolution and multi-spectral Landsat 4–5 Thematic Mapper (TM), Landsat 7
Enhanced Thematic Mapper Plus (ETM+) and Landsat 8 Operational Land Imager (OLI) sensors
were utilised for this study. The different ranges of frequencies along with the electromagnetic (EM)
spectrum for TM, ETM+ and OLI are summarised in Table A1. Extra information UCL Open: Environment is an open
scholarship publication, all previous
versions and open peer review
reports can be found online in the
UCL Open: Environment Preprint
server at ucl.scienceopen.com [111] United States Geological Survey (USGS). Landsat 7 (L7)
Data Users Handbook. LSDS-1927 Version 2.0 [online]. Sioux Falls, South Dakota: EROS; 2019 [Accessed 16
December 2021]. Available from: https://prd-wret.s3.us-
west-2.amazonaws.com/assets/palladium/production/
atoms/files/LSDS1927_L7_Data_Users_Handbook-v2. pdf. [99]
Polidoro BA, Carpenter KE, Collins L, Duke NC, Ellison
AM, Ellison JC, et al. The loss of species: mangrove
extinction risk and geographic areas of global concern. PLoS One. 2010;5(4):e10095. 26 / 29 26 / 29 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines UCL OPEN ENVIRONMENT https://doi.org/10.14324/111.444/ucloe.000057 The empty cells correspond to the unavailability of the sensor for a particular feature. ‘B’ represents the band
number and the corresponding wavelength range, enclosed in a parenthesis, and in a micrometre unit. Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines For satellite sensors, the multi-spectral Landsat 4–5 is denoted by ‘TM’, the Landsat 7 Enhanced Thematic Mapper Plus is denoted by ‘ETM+’ and
‘OLI’ stands for Landsat 8 Operational Land Imager. The spatial resolution for each satellite image is denoted by ‘SRes’. ‘WRS’ means worldwide refer-
ence system, indicated in path ‘P’ and row ‘R’. Mangrove forests projection and model’s accuracy Based on the calculation of the transition probabilities of one system at time t2 with the state of the
system at time t1 according to the specific year [108–110], the Markov’s transition probability matrix
was generated for the two time periods, 1988–1993 and 2013–2020 (Table A3). Table A1. Summary of band designations and spatial resolution for TM, ETM+ and OLI [111]
Sensor
Landsat 4–5 TM
Landsat 7 ETM+
Landsat 8 OLI
Spatial resolution
Coastal aerosol
–
–
B1 (0.43–0.45)
30 m
Blue
B1 (0.45–0.52)
B1 (0.45–0.52)
B2 (0.45–0.51)
30 m
Green
B2 (0.52–0.60)
B2 (0.52–0.60)
B3 (0.53–0.59)
30 m
Red
B3 (0.63–0.69)
B3 (0.63–0.69)
B4 (0.64–0.67)
30 m
NIR
B4 (0.76–0.90)
B4 (0.77–0.90)
B5 (0.85–0.88)
30 m
SWIR 1
B5 (1.55–1.75)
B5 (1.55–1.75)
B6 (1.57–1.65)
30 m
SWIR 2
B7 (2.08–2.35)
B7 (2.09–2.35)
B7 (2.11–2.29)
30 m
Thermal
B6 (10.40–12.50)
B6 (10.40–12.50)
B10 (10.60–11.19)
30 m
–
–
B11 (11.50–12.51)
–
Pan-chromatic
–
B8 (0.52–0.90)
B8 (0.50–0.68)
15 m
Cirrus
–
–
B9 (1.36–1.38)
30 m le A1. Summary of band designations and spatial resolution for TM, ETM+ and OLI [111] The empty cells correspond to the unavailability of the sensor for a particular feature. ‘B’ represents the band
number and the corresponding wavelength range, enclosed in a parenthesis, and in a micrometre unit. 27 / 29 UCL OPEN ENVIRONMENT https://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Table A2. Details of acquired Landsat satellite data selected for this study
Satellite sensor
Acquisition date (mm/dd/yy)
SRes (m)
WRS P/R
Satellite sensor
Acquisition date (mm/dd/yy)
SRes (m)
WRS P/R
TM
03/12/1988
30
115/053
ETM+
01/14/2003
30, 15
118/054
TM
01/31/1988
30
116/052
ETM+
01/23/2008
30, 15
115/053
TM
04/20/1988
30
116/053
ETM+
04/19/2008
30, 15
116/052
TM
06/30/1988
30
117/053
ETM+
10/12/2008
30, 15
116/053
TM
09/18/1988
30
117/054
ETM+
04/10/2008
30, 15
117/053
TM
01/29/1988
30
118/054
ETM+
10/03/2008
30, 15
117/054
TM
11/05/1993
30
115/053
ETM+
04/01/2008
30, 15
118/054
TM
12/14/1993
30
116/052
ETM+
10/19/2013
30, 15
115/053
TM
05/20/1993
30
116/053
ETM+
02/28/2013
30, 15
116/052
TM
10/27/1993
30
116/053
ETM+
05/19/2013
30, 15
116/053
TM
07/14/1993
30
117/053
ETM+
03/07/2013
30, 15
117/053
TM
06/12/1993
30
117/054
ETM+
06/27/2013
30, 15
117/054
TM
03/15/1993
30
118/054
ETM+
05/01/2013
30, 15
118/054
TM
11/10/1993
30
118/054
OLI
12/12/2013
30, 15
115/053
TM
01/03/1998
30
115/053
OLI
08/29/2018
30, 15
116/052
TM
03/31/1998
30
116/052
OLI
02/18/2018
30, 15
116/053
TM
03/31/1998
30
116/053
OLI
04/30/2018
30, 15
117/053
TM
01/17/1998
30
117/053
OLI
12/10/2018
30, 15
117/054
TM
01/17/1998
30
117/054
OLI
04/05/2018
30, 15
118/054
TM
02/09/1998
30
118/054
OLI
04/05/2020
30, 15
115/053
ETM+
04/15/2003
30, 15
115/053
OLI
09/19/2020
30, 15
116/052
ETM+
02/17/2003
30, 15
116/052
OLI
09/19/2020
30, 15
116/053
ETM+
02/01/2003
30, 15
116/053
OLI
08/25/2020
30, 15
117/053
ETM+
03/12/2003
30, 15
117/053
OLI
08/25/2020
30, 15
117/054
ETM+
04/13/2003
30, 15
117/054
OLI
05/12/2020
30, 15
118/054 Table A2. Details of acquired Landsat satellite data selected for this study 28 / 29 UCL OPEN ENVIRONMENT Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines https://doi.org/10.14324/111.444/ucloe.000057 Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines Table A3. Calculated transitional probabilities during 1988–2020
Time period
Probability matrix
Mangrove forests
Non-mangrove areas
1988–1993
Mangrove forests
0.531
0.469
Non-mangrove areas
0.401
0.599
2013–2020
Mangrove forests
0.548
0.452
Non-mangrove areas
0.633
0.367
0%
0%
1988
1993
1998
2003
2008
2013
2018
2020
Year
1988
1993
1998
2003
2008
2013
2018
2020
Year
90.55%
88.44%
88.20%
90.75%
91.34%
93.27%
92.88%
92.53%
88.29%
87.05%
89.97%
87.41%
90.03%
89.92%
87.65%
89.86%
0%
2%
4%
6%
8%
10%
12%
14%
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Percentage of error
Percentage of accuracy
Producer's accuracy (PA)
PA MF
PA NMA
OE MF
OE NMA
91.12%
86.66%
90.60%
92.31%
92.87%
91.66%
92.38%
94.90%
0.86
0.73
0.85
0.88
0.86
0.9
0.91
0.93
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
1988
1993
1998
2003
2008
2013
2018
2020
Percentage of accuracy
Year
Overall accuracy and kappa coefficient
Overall accuracy
Kappa coefficient
Figure A1
Classification error matrix of the Landsat
TM, ETM+ and OLI data for multiple
years, 1988, 1993, 1998, 2003, 2008,
2013, 2018 and 2020. The ground
reference data used was the 2010 map
derived from NAMRIA. The mangrove
forests class is donated by ‘MF’ while the
class of non-mangrove areas is denoted
by ‘NMA’. Additionally, the measure
of commission error (type 1 error) is
denoted by ‘CE’ while the omission
error (type 2 error) is denoted by ‘OE’,
respectively. 1988
1993
1998
2003
2008
2013
2018
2020
Year
90.55%
88.44%
88.20%
90.75%
91.34%
93.27%
92.88%
92.53%
88.29%
87.05%
89.97%
87.41%
90.03%
89.92%
87.65%
89.86%
0%
2%
4%
6%
8%
10%
12%
14%
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Percentage of error
Percentage of accuracy
Producer's accuracy (PA)
PA MF
PA NMA
OE MF
OE NMA Overall accuracy and kappa coefficient 91.12%
86.66%
90.60%
92.31%
92.87%
91.66%
92.38%
94.90%
0.86
0.73
0.85
0.88
0.86
0.9
0.91
0.93
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
1988
1993
1998
2003
2008
2013
2018
2020
Percentage of accuracy
Year
Overall accuracy and kappa coefficient
Overall accuracy
Kappa coefficient Calculated transitional probabilities during 1988–2020
Time period
Probability matrix
Mangrove forests
Non-mangrove
1988–1993
Mangrove forests
0.531
0.469
Non-mangrove areas
0.401
0.599
2013–2020
Mangrove forests
0.548
0.452
Non-mangrove areas
0.633
0.367
91.22%
86.04%
92.08%
95.76%
94.28%
93.62%
94.45%
93.83%
89.91%
85.65%
88.76%
87.53%
89.47%
86.98%
90.32%
91.11%
0%
2%
4%
6%
8%
10%
12%
14%
16%
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
1988
1993
1998
2003
2008
2013
2018
2020
Percentage of error
Percentage of accuracy
Year
1988
1993
1998
2003
2008
2013
2018
2020
Year
User's accuracy (UA)
UA MF
UA NMA
CE MF
CE NMA
90.55%
88.44%
88.20%
90.75%
91.34%
93.27%
92.88%
92.53%
88.29%
87.05%
89.97%
87.41%
90.03%
89.92%
87.65%
89.86%
0%
2%
4%
6%
8%
10%
12%
14%
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Percentage of error
Percentage of accuracy
Producer's accuracy (PA)
PA MF
PA NMA
OE MF
OE NMA
91.12%
86.66%
90.60%
92.31%
92.87%
91.66%
92.38%
94.90%
0.86
0.73
0.85
0.88
0.86
0.9
0.91
0.93
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
1988
1993
1998
2003
2008
2013
2018
2020
Percentage of accuracy
Year
Overall accuracy and kappa coefficient
Overall accuracy
Kappa coefficient
A1
ation error matrix of the Landsat
+ and OLI data for multiple
88, 1993, 1998, 2003, 2008,
18 and 2020. The ground
data used was the 2010 map
om NAMRIA. The mangrove
ass is donated by ‘MF’ while the
on-mangrove areas is denoted
. Additionally, the measure
ssion error (type 1 error) is
by ‘CE’ while the omission
e 2 error) is denoted by ‘OE’,
l 91.22%
86.04%
92.08%
95.76%
94.28%
93.62%
94.45%
93.83%
89.91%
85.65%
88.76%
87.53%
89.47%
86.98%
90.32%
91.11%
0%
2%
4%
6%
8%
10%
12%
14%
16%
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
1988
1993
1998
2003
2008
2013
2018
2020
Percentage of error
Percentage of accuracy
Year
User's accuracy (UA)
UA MF
UA NMA
CE MF
CE NMA User's accuracy (UA) 29 / 29
Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
UCL OPEN ENVIRONMENT
https://doi.org/10.14324/111.444/ucloe.000057
Table A3. Figure A1 Classification error matrix of the Landsat
TM, ETM+ and OLI data for multiple
years, 1988, 1993, 1998, 2003, 2008,
2013, 2018 and 2020. The ground
reference data used was the 2010 map
derived from NAMRIA. The mangrove
forests class is donated by ‘MF’ while the
class of non-mangrove areas is denoted
by ‘NMA’. Additionally, the measure
of commission error (type 1 error) is
denoted by ‘CE’ while the omission
error (type 2 error) is denoted by ‘OE’,
respectively. Multi-spatiotemporal analysis of changes in mangrove forests in Palawan, Philippines
UCL OPEN ENVIRONMENT
https://doi.org/10.14324/111.444/ucloe.000057
Table A3. Calculated transitional probabilities during 1988–2020
Time period
Probability matrix
Mangrove forests
Non-mangrove areas
1988–1993
Mangrove forests
0.531
0.469
Non-mangrove areas
0.401
0.599
2013–2020
Mangrove forests
0.548
0.452
Non-mangrove areas
0.633
0.367 Table A3. Calculated transitional probabilities during 1988–2020 29 / 29 29 / 29
|
https://openalex.org/W3112713288
|
https://www.frontiersin.org/articles/10.3389/fphar.2021.646701/pdf
|
English
| null |
Aripiprazole as a candidate treatment of COVID-19 identified through genomic analysis
|
medRxiv (Cold Spring Harbor Laboratory)
| 2,020
|
cc-by
| 6,848
|
Aripiprazole as a Candidate Treatment
of COVID-19 Identified Through
Genomic Analysis Benedicto Crespo-Facorro 1,2*†, Miguel Ruiz-Veguilla 1,2†, Javier Vázquez-Bourgon 2,3,4,
Ana C. Sánchez-Hidalgo 2,5, Nathalia Garrido-Torres 1, Jose M. Cisneros 6,7, Carlos Prieto 8‡
and Jesus Sainz 9‡ Benedicto Crespo-Facorro 1,2*†, Miguel Ruiz-Veguilla 1,2†, Javier Vázquez-Bourgon 2,3,4,
Ana C. Sánchez-Hidalgo 2,5, Nathalia Garrido-Torres 1, Jose M. Cisneros 6,7, Carlos Prieto 8‡
and Jesus Sainz 9‡ 1Department of Psychiatry, School of Medicine, University Hospital Virgen del Rocio-IBIS, Sevilla, Spain, 2Spanish Network for
Research in Mental Health (CIBERSAM), Sevilla, Spain, 3Department of Psychiatry, University Hospital Marques de Valdecilla -
Instituto de Investigacion Marques de Valdecilla (IDIVAL), Santander, Spain, 4Department of Medicine and Psychiatry, School of
Medicine, University of Cantabria, Santander, Spain, 5Seville Biomedical Research Centre (IBiS), Sevilla, Spain, 6Department of
Infectious Diseases, Microbiology and Preventive Medicine, Institute of Biomedicine of Seville, University Hospital Virgen del
Rocio, University of Seville, Salamanca, Spain, 7Spanish Network for Research in Infectious Diseases (REIPI), Madrid, Spain,
8Bioinformatics Service, Nucleus, University of Salamanca, Salamanca, Spain, 9Spanish National Research Council (CSIC),
Institute of Biomedicine and Biotechnology of Cantabria, Santander, Spain ORIGINAL RESEARCH
published: 02 March 2021
doi: 10.3389/fphar.2021.646701 Edited by:
Siddappa N Byrareddy,
University of Nebraska Omaha,
United States Reviewed by:
Sanjay Rathod,
University of Pittsburgh, United States
Subhash Chand,
University of Nebraska
Medical Center, United States
*Correspondence:
Benedicto Crespo-Facorro
benedicto.crespo.sspa@
juntadeandalucia.es
†These authors have contributed
equally to this work
‡These authors share senior
authorship Reviewed by:
Sanjay Rathod,
University of Pittsburgh, United States
Subhash Chand,
University of Nebraska
Medical Center, United States Background: Antipsychotics modulate expression of inflammatory cytokines and
inducible inflammatory enzymes. Elopiprazole (a phenylpiperazine antipsychotic drug in
phase 1) has been characterized as a therapeutic drug to treat SARS-CoV-2 infection in a
repurposing study. We aim to investigate the potential effects of aripiprazole (an FDA
approved phenylpiperazine) on COVID-19-related immunological parameters. *Correspondence:
Benedicto Crespo-Facorro
benedicto.crespo.sspa@
juntadeandalucia.es Methods: Differential gene expression profiles of non-COVID-19 vs. COVID-19 RNA-Seq
samples (CRA002390 project in GSA database) and drug-naïve patients with non-affective
psychosis at baseline and after three months of aripiprazole treatment were identified. An
integrative transcriptomic analyses of aripiprazole effects on differentially expressed genes
in COVID-19 patients was performed. Findings: 82 out the 377 genes (21.7%) with expression significantly altered by
aripiprazole have also their expression altered in COVID-19 patients and in 93.9% of
these genes their expression is reverted by aripiprazole. The number of common genes
with expression altered in both analyses is significantly higher than expected (Fisher’s
Exact Test, two tail; p value 3.2e-11). 11 KEGG pathways were significantly enriched with
genes with altered expression both in COVID-19 patients and aripiprazole medicated non-
affective psychosis patients (p adj<0.05). The most significant pathways were associated
to
immune
responses
and
mechanisms
of
hyperinflammation-driven
pathology
(i.e.,“inflammatory bowel disease (IBD)” (the most significant pathway with a p adj of
0.00021), “Th1 and Th2 cell differentiation” and “B cell receptor signaling pathway”) that
have been also associated with COVID19 clinical outcome. Specialty section:
This article was submitted to
Inflammation Pharmacology,
a section of the journal
Frontiers in Pharmacology
Received: 27 December 2020
Accepted: 08 February 2021
Published: 02 March 2021 INTRODUCTION Antipsychotics suppress expression of inflammatory cytokines
and inducible inflammatory enzymes (i.e., cyclooxygenase) and
microglia
activation
(Dinesh
et
al.,
2020). These
anti-
inflammatory
effects
are
elicited
via
the
reduction
of
proinflammatory cytokines production, modulating monocytes
response through TLR and the inhibition of the microglial
activation by reducing the levels of inducible nitric oxide
synthase (iNOS), IL-1β, IL-6, and TNF-α (Kato et al., 2007;
Obuchowicz et al., 2017). In humans, the immunomodulatory
effect of risperidone
(pyridopyrimidines) and aripiprazole
(marketed phenylpiperazine) has been demonstrated (Juncal-
Ruiz et al., 2018), with aripiprazole demonstrating a greater
anti-inflammatory
effect
on
TNF-α,
IL-13,
IL-17α
and
fractalkine. Thus, the protective effect of phenylpiperazine
marketed antipsychotics (aripiprazole) against a pernicious
cytokine
storm
is
a
hypothesis
that
warrants
further
investigation with the aim of unrevealing new off-label drug to
be use in severe COVID19 patients. The SARS-CoV-2 epidemic has become the greatest challenge
facing medicine today. Infected patients present with a wide range
of clinical severity varying from asymptomatic to fatal condition
(Wu et al., 2020). Advanced age, gender (male) and suffering
comorbidities (diabetes, cardiovascular or chronic respiratory
diseases)
are
risk
factors
for
higher
clinical
severity,
hospitalization rate and death from COVID-19 (Rubino et al.,
2020; Zhou et al., 2020). The presence of these comorbidities may
decrease resilience and lower the ability to tolerate additional
cytokine storm (Mangalmurti and Hunter, 2020). COVID-19 individuals who become critically and fatally ill
seem to experience an indiscriminate and runaway immune
response
with
an
unchecked
systemic
overproduction
of
cytokines and immunological disbalance (Bhaskar et al., 2020;
Manjili et al., 2020; Zeng et al., 2020). COVID-19 related
immunopathogenesis is not understood just as an emergent
cytokine storm but also as an impairment of protective T cell
immunity (Chen et al., 2020). Prevalence and severity of COVID-19 infection in patients
with severe mental disorders have yielded to inconsistent results,
likely due to differences in the methodology utilized in these
investigations. Lee and collaborators (2020) reported that
diagnosis of a mental disorder was not associated with
increased likelihood of SARS-CoV infection, but a slightly
higher risk for severe clinical outcomes (Lee et al., 2020). Wang and colleagues (2021) reported a higher overall risk to
get infected among schizophrenia patients. In a retrospective
epidemiological study, we observed that vulnerable severe mental
disorder individuals on long-acting injectable antipsychotics had
a lower risk of SARS-CoV2 infection and a better outcome after
infection (unpublished data). Citation: Citation:
Crespo-Facorro B, Ruiz-Veguilla M,
Vázquez-Bourgon J,
Sánchez-Hidalgo AC,
Garrido-Torres N, Cisneros JM,
Prieto C and Sainz J (2021)
Aripiprazole as a Candidate Treatment
of COVID-19 Identified Through
Genomic Analysis. Front. Pharmacol. 12:646701. doi: 10.3389/fphar.2021.646701 Interpretation: This exploratory investigation may provide further support to the notion
that a protective effect is exerted by aripiprazole (phenylpiperazine) by modulating the
expression of genes that have shown to be altered in COVID-19 patients. Along with many March 2021 | Volume 12 | Article 646701 1 Frontiers in Pharmacology | www.frontiersin.org Aripiprazole as a Treatment of COVID-19 Crespo-Facorro et al. ongoing studies and clinical trials, repurposing available medications could be of use in
countering SARS-CoV-2 infection, but require further studies and trials. ongoing studies and clinical trials, repurposing available medications could be of use in
countering SARS-CoV-2 infection, but require further studies and trials. Keywords: psychosis, inflammation, immunology, coronavirus, repurposing drugs, elopiprazole, SARS-CoV-2 INTRODUCTION y
There are no FDA-approved antivirals or vaccines for any
coronavirus, including SARS-CoV-2, and current treatments for
COVID-19 are limited to supportive therapies and off-label use
of FDA-approved drugs. Anti-inflammatory drugs such as
dexamethasone have been shown to reduce deaths (Vabret
et al., 2020). The crucial role of NLRP3 inflammasome
activation in the pathogenesis of diseases caused by SARS-
CoVs draws also attention toward potential role of its
inhibitors in the treatment of COVID-19 (van den Berg and
Te Velde, 2020). Wide range of different drug classes, such as
cancer therapeutics, antipsychotics, and antimalarials, seem to
have a beneficial effect against MERS and SARS coronaviruses
(Dyall et al., 2017). Weston et al., (2020) observed that, although
infection
cannot
be
prevented,
chlorpromazine
(typical
antipsychotic
drug)
and
chloroquine
protect
mice from
severe clinical disease from SARS-CoV. Clozapine (atypical
antipsychotic
drug)
has
revealed
to
be
effective
in
suppressing the proinflammatory cytokine expression by
limiting
the
NLRP3
inflammasome
activation
in
vitro
(Giridharan et al., 2020). In the same line as above, Riva and
colleagues (2020) analyzed approximately 12,000 drugs in
clinical-stage or Food and Drug Administration (FDA)-
approved small molecules to identify candidate drugs to treat
COVID-19 and reported that elopiprazole (a never marketed
phenylpiperazine antipsychotic drug) was listed among the 21
most potent compounds to inhibit SARS-CoV infection. Phenyl-piperazine derivatives had proved their utility as an
effective
source
of
antiviral
compounds,
with
different
mechanisms of action, for treatment of human adenovirus
and cytomegalovirus (DNA viruses) (Sanchez-Cespedes et al.,
2016). The aim of the present study was to examine the potential
beneficial effects of aripiprazole (antipsychotic) in COVID-19
infection by: 1.- analyzing the profile of gene expression of drug-
naïve patients with psychosis at baseline and after three months of
treatment with aripiprazole (PAFIP sample); and 2.- comparing
the set of genes with altered expression in COVID-19 patients
(Wuhan sample) with the set of genes modulated by aripiprazole
in drug-naïve schizophrenia patients. RESULTS Integrative transcriptomic analyses of aripiprazole effects on
differentially expressed genes in COVID-19 patients: Laboratory Assessments Blood samples were obtained from 57 fasting non-affective
psychosis subjects (25 males and 32 females; mean age of
31.54 years) from 8:00 to 10:00 a.m. by the same staff and in
the same setting. A detailed description of methodology followed
to assess biochemical variables is available upon request to the
authors. None of the patients had a chronic inflammation or
infection, or were taking medication that could influence the
results of blood tests. FIGURE 1 | Number of genes with expression altered in Wuhan sample
(COVID-19) and PAFIP sample (aripiprazole-treated). analyzed with RaNA-Seq (Prieto and Barrios, 2020) cloud
platform and differential expression genes were detected by
means of DESeq2 (Love et al., 2014) cloud using a Wald test, a
parametric fit type and setting an adjusted p-value cutoff
of 0.01. Patients
Setting and Sample Study The cohort analyzed to study the effect of aripiprazole was
obtained at the University Hospital Marques de Valdecilla
(Cantabria, Spain). Conforming to international standards for
research ethics, this study was approved by the Cantabria
Ethics Institutional Review Board (IRB). Patients meeting March 2021 | Volume 12 | Article 646701 Frontiers in Pharmacology | www.frontiersin.org 2 Aripiprazole as a Treatment of COVID-19 Crespo-Facorro et al. inclusion criteria for a first episode of non-affective psychosis
(drug-naïve)
provided
written
informed
consent
to
be
included in the study. After informed consent was signed,
patients were included in a prospective, randomized, flexible-
dose, open-label study (Crespo-Facorro et al., 2017; Mayoral
Van-Son et al., 2021). FIGURE 1 | Number of genes with expression altered in Wuhan sample
(COVID-19) and PAFIP sample (aripiprazole-treated). FIGURE 1 | Number of genes with expression altered in Wuhan sample
(COVID-19) and PAFIP sample (aripiprazole-treated). RNA Extraction Total RNA was extracted from blood using the Tempus™Blood
RNA Tube and the Tempus™Spin RNA Isolation Kit (Applied
Biosystems, Foster City, CA, United States) following the
manufacturer’s protocols. To select only high-quality RNA, the
RNA
integrity
number
(RIN)
was
characterized
with
a
Bioanalyzer
(Agilent
Technologies,
Santa
Clara,
CA,
United States) and samples with a RIN of at least 7.6 were used. RNA Next-Generation Sequencing q
g
Total RNA was extracted from peripheral blood of each
individual. The messenger RNA (mRNA) obtained from
blood was sequenced at the Centro Nacional de Análisis
Genómico (CNAG) using Illumina HiSeq instruments (San
Diego, CA, United States). The mRNA was isolated from the
total RNA and was fragmented once transformed into
complementary DNA (cDNA). Fragments of 300bp on
average were selected to construct the cDNA libraries for
sequencing. Pair-end sequences of 70 nucleotides for each
end were produced. The mRNA from blood samples of
57 drug-naïve non-affective psychosis patients at baseline
and
after
3 months
of
continuous
treatment
with
aripiprazole was sequenced. COVID-19 Patients Versus Healthy Donors
(Wuhan Sample) Sequence files were aligned to the GRCh38 human reference
genome (Gencode release 25) using the STAR aligner (Harrow
et al., 2012; Dobin et al., 2013). Reads count were normalized with
the Voom algorithm using the cyclic loess method (Law et al.,
2014) and significant gene expression changes between treated
and naïve patients were identified with lima (Smyth et al., 2005). We performed a paired analysis using a Wald test and a
parametric fit type with an adjusted p-value cutoff of 0.01. We found 2,137 genes with significant differential expression
between COVID-19 patients and controls (p adj value < 0.05)
(Supplementary Table S1). The most significant gene was the
Charcot-Leyden crystal galectin gene (CLC) with a p adj value of
7.8e-23. Drug-Naïve Non-affective Psychosis
Patients at Entry and After 3 months of
Aripiprazole Treatment (PAFIP Sample)
We found 377 genes with significant differential expression
before
and
after
medication
(p
adj
value
<
0.05)
(Supplementary Table S2). The two most significant genes
were the LIM domain only 4 (LMO4) and the ATP binding
cassette subfamily A member 9 (ABCA9) with a p adj value of
0.0039. Statistical and Bioinformatic Analysis Statistical significance of differential expression co-occurrence
between COVID-19 and aripiprazole studies was calculated with
a Fisher Exact test. Functional enrichment analyses, to identify
biological pathways in the Biosystem database (downloaded on
January 2020) (Geer et al., 2010) with a significant presence of
differential expressed genes, were carried out with a Fisher Exact
test. KEGG was used for representation and analysis of molecular
networks (Kanehisa et al., 2010). Visualization and final
representation of pathways were performed with the pathview
R package (Luo and Brouwer, 2013). Differential Gene Expression Between
COVID-19 and Aripiprazole-Treated
Samples 82 out the 377 genes (21.7%) modified by aripiprazole treatment
(PAFIP sample) are significantly also altered in COVID-19
patients (Wuhan sample) (Figure 1). The number of common
genes to both analyses is significantly higher than expected by
chance (Fisher’s Exact Test, two tail; p value 3.2e-11). Interestingly, out of the 82 genes with expression altered in
both analyzed cohorts 55 genes have decreased expression after
aripiprazole medication and increased expression in COVID-19
patients; also, out of the 82 genes common to both cohorts 22
have increased expression after aripiprazole medication and
decreased expression in COVID-19 patients. In total 77 genes
out of 82 (93.9%) have altered expression in different direction
when we compared the effects of COVID-19 and aripiprazole
medication. In
more
severe
COVID-19
cases,
death
results
from
hypoxemic respiratory failure in patients developing severe
acute respiratory distress syndrome and is associated, in a
substantial
portion
of
patients,
with
an
inflammatory
syndrome and cytokine storm (Mehta et al., 2020) that may
originate from immune cells (Chen et al., 2020). Longitudinal
analysis of the immune response observed in a fatal case of
COVID-19 revealed waves of a pro-inflammatory cytokine storm,
Th1 and Th2 activation, and markers of T cell exhaustion,
apoptosis, cell cytotoxicity, and endothelial activation were
until the fatal outcome (Bouadma et al., 2020). Wuhan COVID19 Dataset In
more
severe
COVID-19
cases,
death
results
from
hypoxemic respiratory failure in patients developing severe
acute respiratory distress syndrome and is associated, in a
substantial
portion
of
patients,
with
an
inflammatory
syndrome and cytokine storm (Mehta et al., 2020) that may
originate from immune cells (Chen et al., 2020). Longitudinal
analysis of the immune response observed in a fatal case of
COVID-19 revealed waves of a pro-inflammatory cytokine storm,
Th1 and Th2 activation, and markers of T cell exhaustion,
apoptosis, cell cytotoxicity, and endothelial activation were
until the fatal outcome (Bouadma et al., 2020). related
pathways,
including
“Inflammatory
bowel
disease
(IBD)”, “Th1 and Th2 cell differentiation”, “Fc epsilon RI
signaling pathway”, “B cell receptor signaling pathway”, “NF-
kappa B signaling pathway” and “Th17 cell differentiation”. It is
worth to remark that these immunological pathways have been
associated with COVID19 clinical outcome. Thus, a wide array of
host
humoral
and
cellular
immune
response
alterations
associated
with
SARS-CoV-2
infection
might
cause
an
uncontrolled or insufficient immune response that may lead to
immunopathology and cause severe damage to patients. (Tay
et al., 2020; Wang et al., 2021). Lymphopenia marked by T cell
and NK cell dysfunction, increases in proinflammatory markers
and
cytokines,
and
potential
blood
hypercoagulability
characterize severe COVID-19 cases (Vabret et al., 2020). KEGG Pathways Significantly Enriched With
Common Genes With Altered Expression in
COVID-19 and Aripiprazole-Treated
Samples The analysis of pathways for enrichment of common genes with
altered expression in COVID-19 and aripiprazole patients shows
11 pathways significantly enriched (p value Fisher <0.05)
(Table 1). Several of those pathways are related to the
immune system such as the “inflammatory bowel disease
(IBD)” (the most significant pathway; p adj of 0.00021), “Th1
and Th2 cell differentiation” and “B cell receptor signaling
pathway,” both related to the defense against infections. The TH17 type response profoundly also contributes to the
cytokine storm in pulmonary viral infection including SARS-
CoV-2 (Josset et al., 2013). Compared with non-ICU COVID-19
patients, ICU COVID-19 patients have even higher levels of
several cytokines specifically involved in TH17 type responses
(Huang C. et al., 2020). It has been recently proposed that JAK2
inhibitor (Fedratinib@) can prevent the deteriorating outcomes
of TH17 associated cytokine storm in COVID-19 by suppressing
the production of several TH17 signature cytokines (Wu and
Yang, 2020). Targeting the TH17 pathway may benefit the
patients with TH17 dominant immune profiles. Wuhan COVID19 Dataset COVID data was downloaded from the GSA server (Wang
et al., 2017) with the CRA002390 identifier (Xiong et al.,
2020). This research collected RNA-Seq samples of peripheral
blood mononuclear cells (PBMC) from three COVID-19
patients (all were males; mean age of 45.3 years) and three
healthy donors at Zhongnan Hospital of Wuhan University
that were included in the present study. These data were March 2021 | Volume 12 | Article 646701 Frontiers in Pharmacology | www.frontiersin.org 3 Aripiprazole as a Treatment of COVID-19 Crespo-Facorro et al. TABLE 1 | KEGG pathways significantly enriched with genes with altered expression in COVID19 patients and schizophrenia patients treated with aripiprazole. Pathway ID
Source
Pathway name
Observed %
Expected %
p value Fisher
No. of genes per
pathway
Gene symbol
hsa05321
KEGG
Inflammatory bowel
disease (IBD)
8.16
0.57
0.00021
4
GATA3 HLA-DQB1 IL18 STAT4
hsa04658
KEGG
Th1 and Th2 cell differentiation
8.16
0.82
0.00079
4
GATA3 HLA-DQB1 NFATC2
STAT4
hsa04380
KEGG
Osteoclast differentiation
8.16
1.17
0.00281
4
BTK NFATC2 SYK LILRB4
hsa04664
KEGG
Fc epsilon RI signaling pathway
6.12
0.62
0.00382
3
BTK PLA2G4A SYK
hsa04662
KEGG
B cell receptor signaling pathway
6.12
0.65
0.00430
3
BTK NFATC2 SYK
hsa05164
KEGG
Influenza A
8.16
1.50
0.00653
4
HLA-DQB1 HLA-DQB1 IL18
NLRP3
hsa04064
KEGG
NF-kappa B signaling pathway
6.12
0.84
0.00867
3
BTK LTA SYK
hsa04659
KEGG
Th17 cell differentiation
6.12
0.95
0.01202
3
GATA3 HLA-DQB1 NFATC2
hsa04611
KEGG
Platelet activation
6.12
1.09
0.01712
3
BTK PLA2G4A SYK
hsa05166
KEGG
HTLV-I infection
8.16
2.33
0.02787
4
FZD2 HLA-DQB1 LTA NFATC2
hsa05152
KEGG
Tuberculosis
6.12
1.53
0.03986
3
HLA-DQB1 IL18 SYK related
pathways,
including
“Inflammatory
bowel
disease
(IBD)”, “Th1 and Th2 cell differentiation”, “Fc epsilon RI
signaling pathway”, “B cell receptor signaling pathway”, “NF-
kappa B signaling pathway” and “Th17 cell differentiation”. It is
worth to remark that these immunological pathways have been
associated with COVID19 clinical outcome. Thus, a wide array of
host
humoral
and
cellular
immune
response
alterations
associated
with
SARS-CoV-2
infection
might
cause
an
uncontrolled or insufficient immune response that may lead to
immunopathology and cause severe damage to patients. (Tay
et al., 2020; Wang et al., 2021). Lymphopenia marked by T cell
and NK cell dysfunction, increases in proinflammatory markers
and
cytokines,
and
potential
blood
hypercoagulability
characterize severe COVID-19 cases (Vabret et al., 2020). DISCUSSION Our results herein revealed that the differently expressed genes in
COVID-19 patients and schizophrenia patients treated with
aripiprazole were highly associated with numerous immune- Among biologic agents in patients with severe COVID-19
inhibiting Fc epsilon RI signaling has been proposed (Yalcin and
Yalcin, 2021). Omalizumab specifically binds to the CH3 domain, is March 2021 | Volume 12 | Article 646701 Frontiers in Pharmacology | www.frontiersin.org 4 Aripiprazole as a Treatment of COVID-19 Crespo-Facorro et al. FIGURE 2 | Chemical structures of two phenypiperazines with
antipsychotic effects: elopiprazole and aripiprazole. near to the binding site for the high affinity IgE Fc receptors type-I
(also called FceRI) of human IgE (Metz et al., 2017). A critical issue
in patients with Covid-19 is the viremia and the overresponse to this
viremia with increase of ferritin, CRP and D-Dimer that are directly
associated with the mortality (Huang I. et al., 2020). It has been
described that omalizumab safely decreases the coagulant proteins
(D-Dimer) and proinflammatory cytokines/mediators and increases
the anti-coagulant proteins (protein C, S) in patients with sepsis
(Yalcin et al., 2013; Criado et al., 2020). So, it could be anticipated
that we may administer it for severe COVID-19 (Yalcin and Yalcin,
2021). We observed that those genes differentially expressed in
COVID-19 and aripiprazole-treated patients were involved in
Inflammatory bowel disease (IBD). Autoinflammatory diseases
(IBD) that present with bowel inflammation and intractable
diarrhea owing to an inappropriate inflammatory response,
with also altered key immune pathways underlying persistent
inflammation such as excessive IL-1 signaling, constitutive NF-κB
activation, and chronic type I IFN signaling (de Jesus et al., 2015). It is also of interest that JAK-STAT pathways, as well as in
COVID19, play an important role in the inflammatory response
characteristic of IBD and represent a promising therapeutic target
for treatment of the disease. (Pedersen et al., 2014). FIGURE 2 | Chemical structures of two phenypiperazines with
antipsychotic effects: elopiprazole and aripiprazole. phenylpiperazines, containing a phenylpiperazine skeleton, which
consists of a piperazine bound to a phenyl group (Figure 2). In our study, differentially expressed genes involved in NF-
kappa B signaling pathway were identified according to KEGG
pathway analysis. The activated NF-κB transcription factors serve
as a “rapid acting” primary transcription factor regulating diverse
cellular responses associated with chronic inflammatory states,
septic shock syndrome and multiorgan failure (Zhang et al.,
2017). DISCUSSION Hyper-activation of the nuclear factor kappa-light-chain
enhancer of activated B cells (NF-κB) pathway has been
implicated in the pathogenesis of the severe/critical COVID19
phenotype (Hirano and Murakami, 2020). Cromolyn, which
inhibit
NF-κB
mediated
cytokine
production,
has
been
suggested as a repurposing drug in the fight against COVID-
19 (Karadsheh Adli, 2020). Many of the drugs currently effective
in COVID disease appear to have links to the NF-κB cascade of
immune regulation (Hariharan et al., 2020). If our results herein prove to be certain one may expect a milder
severity
of
COVID-19
in
patients
on
aripiprazole
(phenylpiperazine). Differences
in
the
definitions
and
methodology utilized in recent investigations have yielded to
inconsistent results about the prevalence and severity of COVID-
19 infection among patients with severe mental disorders (Lee et al.,
2020; Wang et al., 2021). It is of worth to pinpoint that Wang and
colleagues (2021) reported that chronic schizophrenia patients have
a lower association compared with recent diagnosis of schizophrenia
(less than 1 year) (AOR 1.48, 95% CI: 1.33–1.65 vs. AOR 9.89,
95% CI: 8.68–11.26), giving place to the speculation that
antipsychotic treatment may exert a protective effect. Nemani
et al., (2021) recently described that schizophrenia spectrum
diagnosis was associated with an increase of death or discharge
to hospice outcome at 45 days following testing and highlighted the
relevance
of
evaluating
the
potential
protective
effects
of
psychotropic medications. Our data retrospectively exploring an
epidemiological sample of people with severe mental disorders who
are on long-acting injectable antipsychotics revealed a lower risk of
SARS-CoV2 infection and a better outcome after infection in this
group of antipsychotic treated individuals compared to overall
COVID-19 people (Canal-Rivero et al., 2021). Despite all recent generated scientific knowledge, immune
responses
and
mechanisms
of
hyperinflammation-driven
pathology need to be further elucidated to address how these
immune differences across patients or between different types of
coronavirus infections dictate who succumbs to disease and who
remains asymptomatic (Vabret et al., 2020). The fact that antipsychotics have a demonstrated effect on
immunological
pathways,
inducible
inflammatory
enzymes
(i.e., cyclooxygenase), and microglia activation (Dinesh et al., 2020)
may lead to the speculation about the beneficial effects of these drugs
on controlling the acute hyper-inflammatory response that may be
responsible for critical COVID19 illness. A recent study (Riva et al.,
2020) profiled a library of approximately 12,000 drugs in clinical-stage
or FDA-approved small molecules to identify candidate drugs to treat
COVID-19. DISCUSSION In the list of the 21 most potent compounds to inhibit
infection validated in dose response across multiple cell lines there are
two antipsychotic drugs, elopiprazole and 8-(3-Chlorostyryl) caffeine
which are in phase I and preclinical stages respectively. Elopiprazole
and aripiprazole belong to the class of organic compounds known as Some limitations need to be taken into when interpreting our
results. First, differences in gender distribution between cohorts
may limit the comparability of gene expression data. Second, the
pattern of gene expression associated to COVID19 infection
raises from three COVID19 patients and three healthy donors
from whom there is limited access to subjects’ characteristics. Frontiers in Pharmacology | www.frontiersin.org REFERENCES Crespo-Facorro, B., Ruiz-Veguilla, M., Vázquez-Bourgon, J., Sánchez-Hidalgo, A. C., Garrido-Torres, N., Cisneros, J. M., et al. (2020). Aripiprazole as a candidate
treatment of COVID-19 identified through genomic analysis. MedRxiv. doi:10. 1101/2020.12.05.20244590 Bhaskar, S., Sinha, A., Banach, M., Mittoo, S., Weissert, R., Kass, J. S., et al. (2020). Cytokine storm in COVID-19-Immunopathological mechanisms, clinical
considerations, and therapeutic approaches: the REPROGRAM consortium
position paper. Front. Immunol. 11, 1648. doi:10.3389/fimmu.2020.01648 Criado, P. R., Pagliari, C., Criado, R. F. J., Marques, G. F., and Belda, W. (2020). What the physicians should know about mast cells, dendritic cells, urticaria, and
omalizumab during COVID-19 or asymptomatic infections due to SARS-CoV-
2? Dermatol. Ther. 33 (6), e14068. doi:10.1111/dth.14068 Bouadma, L., Wiedemann, A., Patrier, J., Surénaud, M., Wicky, P. H., Foucat, E.,
et al. (2020). Immune alterations in a patient with SARS-CoV-2-related acute
respiratory distress syndrome. J. Clin. Immunol. 40 (8), 1082–1092. doi:10. 1007/s10875-020-00839-x de Jesus, A. A., Canna, S. W., Liu, Y., and Goldbach-Mansky, R. (2015). Molecular
mechanisms in genetically defined autoinflammatory diseases: disorders of
amplified danger signaling. Annu. Rev. Immunol. 33, 823–874. doi:10.1146/
annurev-immunol-032414-112227 Canal-Rivero, M., Barragán, R. C., García, A. R., Garrido-Torres, N., Crespo-
Facorro, B., and Ruiz-Veguilla, M. (2021). Lower risk of SARS-CoV2 infection
in individuals with severe mental disorders on antipsychotic treatment: A
retrospective epidemiological study in a representative Spanish population. Schizophrenia Res. 229, 53–54. doi:10.1016/j.schres.2021.02.002 Dinesh, A. A., Islam, J., Khan, J., Turkheimer, F., and Vernon, A. C. (2020). Effects
of antipsychotic drugs: cross talk between the nervous and innate immune
system. CNS Drugs 34, 1229. doi:10.1007/s40263-020-00765-x Dobin, A., Davis, C. A., Schlesinger, F., Drenkow, J., Zaleski, C., Jha, S., et al. (2013). STAR: ultrafast universal RNA-seq aligner. Bioinformatics 29 (1), 15–21. doi:10. 1093/bioinformatics/bts635 Chen, G., Wu, D., Guo, W., Cao, Y., Huang, D., Wang, H., et al. (2020). Clinical and
immunological features of severe and moderate coronavirus disease 2019. J. Clin. Invest. 130 (5), 2620–2629. doi:10.1172/JCI137244 Dyall, J., Gross, R., Kindrachuk, J., Johnson, R. F., Olinger, G. G., Hensley, L. E.,
et al. (2017). Middle East respiratory syndrome and severe acute respiratory
syndrome: current therapeutic options and potential targets for novel therapies. Drugs 77 (18), 1935–1966. doi:10.1007/s40265-017-0830-1 Crespo-Facorro, B., Ortiz-Garcia de la Foz, V., Suarez-Pinilla, P., Valdizan, E. M., Pérez-Iglesias, R., Amado-Señaris, J. A., et al. (2017). Effects of
aripiprazole, quetiapine and ziprasidone on plasma prolactin levels in
individuals with first episode nonaffective psychosis: analysis of a
randomized open-label 1year study. ETHICS STATEMENT The studies involving human participants were reviewed and
approved by the Cantabria Ethics Institutional Review Board
(IRB). The patients/participants provided their written informed
consent to participate in this study. FUNDING and that another never marked phenylpiperazine antipsychotic
(elopiprazole) has been validated as potential treatment for
COVID19, it may be suggested that aripiprazole might be
used as treatment for COVID19. Along with many ongoing
studies and clinical trials, repurposing available medications
could be of use in countering SARS-CoV-2 infection, but
clearly require further studies and trials. The present study was part of a larger prospective longitudinal
study, the “First Episode Psychosis Clinical Program 10”
(PAFIP10) study. ClinicalTrials.gov Identifiers: NCT02200588,
NCT03481465, and NCT03476473. No pharmaceutical industry
or institutional sponsors participated in the study conception and
design, data collection, analysis and interpretation of the results, or
drafting of the manuscript. This work was supported by: SAF2016-
76046-R and SAF2013-46292-R (MINECO and FEDER) to B.C.F. The present study was part of a larger prospective longitudinal
study, the “First Episode Psychosis Clinical Program 10”
(PAFIP10) study. ClinicalTrials.gov Identifiers: NCT02200588,
NCT03481465, and NCT03476473. No pharmaceutical industry
or institutional sponsors participated in the study conception and
design, data collection, analysis and interpretation of the results, or
drafting of the manuscript. This work was supported by: SAF2016-
76046-R and SAF2013-46292-R (MINECO and FEDER) to B.C.F. AUTHOR CONTRIBUTIONS Study
concept
and
design:
BC-F,
MR-V,
CP,
and
JS;
acquisition, analysis or interpretation of data: all authors;
drafting of the manuscript: all authors; critical revision of
the
manuscript
for
important
intellectual
content:
all
authors; obtained funding: BC-F and JS; study supervision:
BC-F, MR-V, CP, and JS. ACKNOWLEDGMENTS We are highly indebted to the participants and their families for
their cooperation in this study. We also thank IDIVAL biobank
(Inés Santiuste and Jana Arozamena) for clinical samples and
data as well as the PAFIP members (Marga Corredera) for the
data collection. We kindly thank all clinical staff at the Hospital
Universitario Virgen del Rocio for support to collect clinical
records and provide clinical care to COVID-19 patients. We also
kindly thank Dra. Marisa Barrigon for helpful discussions
regarding
clinical
data
analysis,
and
Idalino
Rocha
for
manuscript editing and formatting. This manuscript has been
released as a pre-print at medRxiv. Available at: https://doi.org/
10.1101/2020.12.05.20244590 (Crespo-Facorro et al., 2020). SUPPLEMENTARY MATERIAL The Supplementary Material for this article can be found online at:
https://www.frontiersin.org/articles/10.3389/fphar.2021.646701/
full#supplementary-material. DATA AVAILABILITY STATEMENT The datasets presented in this study can be found in online
repositories. The names of the repository/repositories and
accession number(s) can be found below: https://www.ebi.ac. uk/ena, PRJEB42627. CONCLUSION Given that the effect of aripiprazole in gene expression mainly
revert the changes in expression caused by COVID19 infection, March 2021 | Volume 12 | Article 646701 Frontiers in Pharmacology | www.frontiersin.org 5 Aripiprazole as a Treatment of COVID-19 Crespo-Facorro et al. REFERENCES Genome Biol. 15
(2), R29. doi:10.1186/gb-2014-15-2-r29 Wang, Q., Xu, R., and Volkow, N. D. (2021). Increased risk of COVID -19 infection
and mortality in people with mental disorders: analysis from electronic health
records in the United States. World Psychiatry 20 (1), 124–130. doi:10.1002/
wps.20806 Lee, S. W., Yang, J. M., Moon, S. Y., Yoo, I. K., Ha, E. K., Kim, S. Y., et al. (2020). Association between mental illness and COVID-19 susceptibility and clinical
outcomes in South Korea: a nationwide cohort study. Lancet Psychiatry 7 (12),
1025–1031. doi:10.1016/S2215-0366(20)30421-1 Wang, Y., Song, F., Zhu, J., Zhang, S., Yang, Y., Chen, T., et al. 2017). GSA: genome
sequence archive *. Genomics Proteomics Bioinformatics 15 (1), 14–18. doi:10. 1016/j.gpb.2017.01.001 Love, M. I., Huber, W., and Anders, S. (2014). Moderated estimation of fold change
and dispersion for RNA-seq data with DESeq2. Genome Biol. 15 (12), 550. doi:10.1186/s13059-014-0550-8 Weston, S., Coleman, C. M., Haupt, R., Logue, J., Matthews, K., Li, Y., et al. (2020). Broad anti-coronavirus activity of Food and drug administration-approved
drugs against SARS-CoV-2 in vitro and SARS-CoV in vivo. J. Virol. 94 (21),
e01218-20. doi:10.1128/JVI.01218-20 Luo, W., and Brouwer, C. (2013). Pathview: an R/Bioconductor package for
pathway-based data integration and visualization. Bioinformatics 29 (14),
1830–1831. doi:10.1093/bioinformatics/btt285 Mangalmurti, N., and Hunter, C. A. (2020). Cytokine storms: understanding
COVID-19. Immunity 53 (1), 19–25. doi:10.1016/j.immuni.2020.06.017 Wu, C., Chen, X., Cai, Y., Xia, J., Zhou, X., Xu, S., et al. 2020). Risk factors associated
with acute respiratory distress syndrome and death in patients with coronavirus
disease 2019 pneumonia in Wuhan, China. JAMA Intern. Med. 180 (7),
934–943. doi:10.1001/jamainternmed.2020.0994 Manjili, R. H., Zarei, M., Habibi, M., and Manjili, M. H. (2020). COVID-19 as an
acute inflammatory disease. J. Immunol. 205 (1), 12–19. doi:10.4049/jimmunol. 2000413 Wu, D., and Yang, X. O. (2020). TH17 responses in cytokine storm of COVID-19:
an emerging target of JAK2 inhibitor Fedratinib. J. Microbiol. Immunol. Infect. 53 (3), 368–370. doi:10.1016/j.jmii.2020.03.005 Mayoral Van-Son, J., Gómez-Revuelta, M., Ayesa-Arriola, R., Vázquez-Bourgón, J.,
Foz, V. O.-G. de. la., Ruiz-Veguilla, M., et al. (2021). Comparison of aripiprazole
and risperidone effectiveness in first episode non-affective psychosis: rationale
and design of a prospective, randomized, 3-phase, investigator-initiated study
(PAFIP-3). Rev. Psiquiatr. Salud Ment. doi:10.1016/j.rpsm.2021.01.004 Xiong, Y., Liu, Y., Cao, L., Wang, D., Guo, M., Jiang, A., et al. (2020). Transcriptomic
characteristics
of
bronchoalveolar
lavage
fluid
and
peripheral blood mononuclear cells in COVID-19 patients. Emerg. Microbes
Infect. 9 (1), 761–770. REFERENCES Schizophr. Res. 189, 134–141. doi:10.1016/j.schres.2017.01.046 Drugs 77 (18), 1935–1966. doi:10.1007/s40265-017-0830-1 Geer, L. Y., Marchler-Bauer, A., Geer, R. C., Han, L., He, J., He, S., et al. (2010). The
NCBI BioSystems database. Nucleic Acids Res. 38, D492–D496. doi:10.1093/
nar/gkp858 March 2021 | Volume 12 | Article 646701 Frontiers in Pharmacology | www.frontiersin.org 6 Aripiprazole as a Treatment of COVID-19 Crespo-Facorro et al. with a reduction of FcεRI-positive cells in the skin. Theranostics 7 (5),
1266–1276. doi:10.7150/thno.18304 Giridharan, V. V., Scaini, G., Colpo, G. D., Doifode, T., Pinjari, O. F., Teixeira, A. L.,
et al. (2020). Clozapine prevents poly (I:C) induced inflammation by
modulating NLRP3 pathway in microglial cells. Cells 9 (3), 577. doi:10.3390/
cells9030577 with a reduction of FcεRI-positive cells in the skin. Theranostics 7 (5),
1266–1276. doi:10.7150/thno.18304 Nemani, K., Li, C., Olfson, M., Blessing, E. M., Razavian, N., Chen, J., et al. (2021). Association of psychiatric disorders with mortality among patients with
COVID-19. JAMA Psychiatry. doi:10.1001/jamapsychiatry.2020.4442 Hariharan, A., Hakeem, A. R., Radhakrishnan, S., Reddy, M. S., and Rela, M. (2020). The role and therapeutic potential of NF-kappa-B pathway in severe
COVID-19 patients. Inflammopharmacol., 1–10. doi:10.1007/s10787-020-
00773-9 Obuchowicz, E., Bielecka-Wajdman, A. M., Paul-Samojedny, M., and Nowacka, M. (2017). Different influence of antipsychotics on the balance between pro- and
anti-inflammatory cytokines depends on glia activation: an in vitro study. Cytokine 94, 37–44. doi:10.1016/j.cyto.2017.04.004 Harrow, J., Frankish, A., Gonzalez, J. M., Tapanari, E., Diekhans, M., Kokocinski,
F., et al. 2012). GENCODE: the reference human genome annotation for the
ENCODE Project. Genome Res. 22 (9), 1760–1774. doi:10.1101/gr.135350.111 Pedersen, J., Coskun, M., Soendergaard, C., Salem, M., and Nielsen, O. H. (2014). Inflammatory pathways of importance for management of inflammatory
bowel disease. World J. Gastroenterol. 20 (1), 64–77. doi:10.3748/wjg.v20. i1.64 Hirano, T., and Murakami, M. (2020). COVID-19: a new virus, but a familiar
receptor and cytokine release syndrome. Immunity 52 (5), 731–733. doi:10. 1016/j.immuni.2020.04.003 Prieto, C., and Barrios, D. (2020). RaNA-Seq: interactive RNA-Seq analysis from
FASTQ files to functional analysis. Bioinformatics 36 (6), 1955–1956. doi:10. 1093/bioinformatics/btz854 Huang, C., Wang, Y., Li, X., Ren, L., Zhao, J., Hu, Y., et al. (2020). Clinical features
of patients infected with 2019 novel coronavirus in Wuhan, China. Lancet 395
(10223), 497–506. doi:10.1016/S0140-6736(20)30183-5 Riva, L., Yuan, S., Yin, X., Martin-Sancho, L., Matsunaga, N., Pache, L., et al. (2020). Discovery of SARS-CoV-2 antiviral drugs through large-scale
compound repurposing. Nature 586 (7827), 113–119. doi:10.1038/s41586-
020-2577-1 Huang, I., Pranata, R., Lim, M. REFERENCES A., Oehadian, A., and Alisjahbana, B. (2020). C-reactive protein, procalcitonin, D-dimer, and ferritin in severe coronavirus
disease-2019: a meta-analysis. Ther. Adv. Respir. Dis. 14, 1753466620937175. doi:10.1177/1753466620937175 Rubino, S., Kelvin, N., Bermejo-Martin, J. F., and Kelvin, D. (2020). As COVID-19
cases, deaths and fatality rates surge in Italy, underlying causes require
investigation. J. Infect. Dev. Ctries. 14 (3), 265–267. doi:10.3855/jidc.12734 Josset, L., Menachery, V. D., Gralinski, L. E., Agnihothram, S., Sova, P., Carter, V. S., et al. (2013). Cell host response to infection with novel human coronavirus
EMC predicts potential antivirals and important differences with SARS
coronavirus. MBio 4 (3), e00165–13. doi:10.1128/mBio.00165-13 Sánchez-Céspedes, J., Martínez-Aguado, P., Vega-Holm, M., Serna-Gallego, A.,
Candela, J. I., Marrugal-Lorenzo, J. A., et al. (2016). New 4-Acyl-1-
phenylaminocarbonyl-2-phenylpiperazine derivatives as potential inhibitors
of adenovirus infection. Synthesis, biological evaluation, and structure-
activity relationships. J. Med. Chem. 59 (11), 5432–5448. doi:10.1021/acs. jmedchem.6b00300 Juncal-Ruiz, M., Riesco-Dávila, L., Ortiz-García de la Foz, V., Martínez-Garcia, O.,
Ramírez-Bonilla, M., Ocejo-Viñals, J. G., et al. (2018). Comparison of the anti-
inflammatory effect of aripiprazole and risperidone in 75 drug-naïve first
episode psychosis individuals: a 3 months randomized study. Schizophr. Res. 202, 226–233. doi:10.1016/j.schres.2018.06.039 Smyth, G. K., Michaud, J., and Scott, H. S. (2005). Use of within-array replicate
spots
for assessing differential expression
in microarray
experiments. Bioinformatics 21 (9), 2067–2075. doi:10.1093/bioinformatics/bti270 Kanehisa, M., Goto, S., Furumichi, M., Tanabe, M., and Hirakawa, M. (2010). KEGG for representation and analysis of molecular networks involving diseases
and drugs. Nucleic Acids Res. 38, D355–D360. doi:10.1093/nar/gkp896 Tay, M. Z., Poh, C. M., Rénia, L., MacAry, P. A., and Ng, L. F. P. (2020). The trinity
of COVID-19: immunity, inflammation and intervention. Nat. Rev. Immunol. 20 (6), 363–374. doi:10.1038/s41577-020-0311-8 Karadsheh Adli, J. (2020). Re: Mahase Elisabeth. Covid-19: What treatments are
being 478 investigated? BMJ 368, m1252, 2020 . Available at https://www.bmj. com/content/368/bmj.m1252/rr-1 Vabret, N., Britton, G. J., Gruber, C., Hegde, S., Kim, J., Kuksin, M., et al. (2020). Immunology of COVID-19: current state of the science. Immunity 52 (6),
910–941. doi:10.1016/j.immuni.2020.05.002 Kato, T., Monji, A., Hashioka, S., and Kanba, S. (2007). Risperidone significantly
inhibits interferon-gamma-induced microglial activation in vitro. Schizophr. Res. 92 (1–3), 108–115. doi:10.1016/j.schres.2007.01.019 van den Berg, D. F., and Te Velde, A. A. (2020). Severe COVID-19: NLRP3
inflammasome dysregulated. Front. Immunol. 11, 1580. doi:10.3389/fimmu. 2020.01580 Law, C. W., Chen, Y., Shi, W., and Smyth, G. K. (2014). voom: precision weights
unlock linear model analysis tools for RNA-seq read counts. REFERENCES doi:10.1080/22221751.2020.1747363 Mehta, P., McAuley, D. F., Brown, M., Sanchez, E., Tattersall, R. S., and Manson,
J. J. (2020). COVID-19:
consider
cytokine
storm
syndromes
and
immunosuppression. Lancet 395 (10229), 1033–1034. doi:10.1016/S0140-
6736(20)30628-0 Yalcin, A. D., Cilli, A., Bisgin, A., Strauss, L. G., and Herth, F. (2013). Omalizumab
is effective in treating severe asthma in patients with severe cardiovascular
complications and its effects on sCD200, d-dimer, CXCL8, 25-hydroxyvitamin
D and IL-1β levels. Expert Opin. Biol. Ther. 13 (9), 1335–1341. doi:10.1517/
14712598.2013.819338 Metz, M., Staubach, P., Bauer, A., Brehler, R., Gericke, J., Kangas, M., et al. (2017). Clinical efficacy of omalizumab in chronic spontaneous urticaria is associated March 2021 | Volume 12 | Article 646701 Frontiers in Pharmacology | www.frontiersin.org 7 Aripiprazole as a Treatment of COVID-19 Crespo-Facorro et al. Yalcin, A. D., and Yalcin, A. N. (2021). Future perspective: biologic agents in
patients with severe COVID-19. Immunopharmacol. Immunotoxicol. 43 (1),
1–7. doi:10.1080/08923973.2020.1818770 educational
events
from
Janssen,
Lundbeck,
and
Otsuka
Pharmaceuticals. JV-B has received unrestricted research funding from
Instituto de Investigación Marqués de Valdecilla (IDIVAL). He has also
received honoraria for his participation as a consultant and/or as a speaker
at educational events from Janssen-Cilag and Lundbeck. JC has received
honoraria as a speaker from Novartis, Astellas Pharma, Pfizer, MSD,
Janssen Pharmaceuticals, and AstraZeneca, outside the submitted work. He has also received report grants from Instituto de Salud Carlos III,
Spanish Government, co-financed by the European Development Regional
Fund “A way to achieve Europe,” during the conduct of the study. educational
events
from
Janssen,
Lundbeck,
and
Otsuka
Pharmaceuticals. JV-B has received unrestricted research funding from
Instituto de Investigación Marqués de Valdecilla (IDIVAL). He has also
received honoraria for his participation as a consultant and/or as a speaker
at educational events from Janssen-Cilag and Lundbeck. JC has received
honoraria as a speaker from Novartis, Astellas Pharma, Pfizer, MSD,
Janssen Pharmaceuticals, and AstraZeneca, outside the submitted work. He has also received report grants from Instituto de Salud Carlos III,
Spanish Government, co-financed by the European Development Regional
Fund “A way to achieve Europe,” during the conduct of the study. Zeng, Z., Yu, H., Chen, H., Qi, W., Chen, L., Chen, G., et al. (2020). Longitudinal
changes of inflammatory parameters and their correlation with disease severity
and outcomes in patients with COVID-19 from Wuhan, China. Crit. Care 24
(1), 525. doi:10.1186/s13054-020-03255-0 Zhang, Q., Lenardo, M. J., and Baltimore, D. (2017). Frontiers in Pharmacology | www.frontiersin.org REFERENCES 30 Years of NF-κB: a
blossoming of relevance to human pathobiology. Cell 168 (1–2), 37–57. doi:10.1016/j.cell.2016.12.012 Zhou, F., Yu, T., Du, R., Fan, G., Liu, Y., Liu, Z., et al. (2020). Clinical course and
risk factors for mortality of adult inpatients with COVID-19 in Wuhan, China:
a retrospective cohort study. Lancet 395 (10229), 1054–1062. doi:10.1016/
S0140-6736(20)30566-3 The remaining authors declare that the research was conducted in the absence of
any commercial or financial relationships that could be construed as a potential
conflict of interest. Conflict of Interest: BC-F has received unrestricted research funding from Conflict of Interest: BC-F has received unrestricted research funding from
Instituto de Salud Carlos III, MINECO, Gobierno de Cantabria, Spanish
Network for Research in Mental Health (CIBERSAM), from the seventh
European Union Framework Program and Lundbeck. He has also received
honoraria for his participation as a consultant and/or as a speaker at
educational events from Janssen Johnson & Johnson, Mylan, Lundbeck,
and Otsuka Pharmaceuticals. MR-V has received unrestricted research
funding from Instituto de Salud Carlos III. He has also received
honoraria for his participation as a consultant and/or as a speaker at Copyright © 2021 Crespo-Facorro, Ruiz-Veguilla, Vázquez-Bourgon, Sánchez-
Hidalgo, Garrido-Torres, Cisneros, Prieto and Sainz. This is an open-access
article distributed under the terms of the Creative Commons Attribution License
(CC BY). The use, distribution or reproduction in other forums is permitted,
provided the original author(s) and the copyright owner(s) are credited and that
the original publication in this journal is cited, in accordance with accepted academic
practice. No use, distribution or reproduction is permitted which does not comply
with these terms. Copyright © 2021 Crespo-Facorro, Ruiz-Veguilla, Vázquez-Bourgon, Sánchez-
Hidalgo, Garrido-Torres, Cisneros, Prieto and Sainz. This is an open-access
article distributed under the terms of the Creative Commons Attribution License
(CC BY). The use, distribution or reproduction in other forums is permitted,
provided the original author(s) and the copyright owner(s) are credited and that
the original publication in this journal is cited, in accordance with accepted academic
practice. No use, distribution or reproduction is permitted which does not comply
with these terms. March 2021 | Volume 12 | Article 646701 Frontiers in Pharmacology | www.frontiersin.org 8
|
https://openalex.org/W3199987258
|
https://apsy.sbu.ac.ir/article_101199_961a4a719f26d0f64ecbb1aa27451cd0.pdf
|
Persian
| null |
Evaluation of the Effectiveness of Intimacy From the Inside Out Relationship.Therapy (IFIO) in Improving a Couple's Secure
|
Faṣlnāmah-i ravān/shināsī-i kārburdī./Faṣlnāmah-i ravān/shināsī-i kārburdī.
| 2,021
|
cc-by
| 9,150
|
Citation: Heydarianfar, N., Aslani, Kh., Rajabi, Gh., & Amanelahi, A. (2021). Evaluation of the
Effectiveness of Intimacy From the Inside Out Relationship.Therapy (IFIO) in Improving a Couple's
Secure. Quarterly of Applied Psychology, 15(2):297-317. 10.52547/APSY.2021.221513.1041
20.1001.1.20084331.1400.15.2.6.3
Research Article
Evaluation of the Effectiveness of Intimacy From the Inside
Out Relationship.Therapy (IFIO) in Improving a Couple's
Secure
N. Heydarianfar1, Kh. Aslani2*, Gh. Rajabi3 & A. Amanelahi2
1. PhD. Student of Counseling, , Shahid Chamran University of Ahvaz, Ahvaz, Iran.
2. Associate Professor, Department of Family Counseling, Shahid Chamran University of Ahvaz, Ahvaz, Iran. Email: kh.aslani@scu.ac.ir
3. Professor, Department of Family Counseling, Shahid Chamran University of Ahvaz, Ahvaz, Iran.
Quarterly of Applied Psychology, Vol. 15, No. 2 (58), Summer 2021, 297-317
ISSN: 2008-4331- eISSN: 2645- 3541 10.52547/APSY.2021.221513.1041
20.1001.1.20084331.1400.15.2.6.3
Research Article
Evaluation of the Effectiveness of Intimacy From the Inside
Out Relationship.Therapy (IFIO) in Improving a Couple's
Secure
N. Heydarianfar1, Kh. Aslani2*, Gh. Rajabi3 & A. Amanelahi2
1. PhD. Student of Counseling, , Shahid Chamran University of Ahvaz, Ahvaz, Iran. 2. Associate Professor, Department of Family Counseling, Shahid Chamran University of Ahvaz, Ahvaz, Iran. Email: kh.aslani@scu.ac.ir
3. Professor, Department of Family Counseling, Shahid Chamran University of Ahvaz, Ahvaz, Iran. Quarterly of Applied Psychology, Vol. 15, No. 2 (58), Summer 2021, 297-317
ISSN: 2008-4331- eISSN: 2645- 3541 arterly of Applied Psychology, Vol. 15, No. 2 (58), Summer 2021, 297-317 Quarterly of Applied Psychology, Vol. 15, No. 2 (58), Summer 2021, 297-317 Secure
N. Heydarianfar1, Kh. Aslani2*, Gh. Rajabi3 & A. Amanelahi2
1. PhD. Student of Counseling, , Shahid Chamran University of Ahvaz, Ahvaz, Iran. 2. Associate Professor, Department of Family Counseling, Shahid Chamran University of Ahvaz, Ahvaz, Iran. Email: kh.aslani@sc
3. Professor, Department of Family Counseling, Shahid Chamran University of Ahvaz, Ahvaz, Iran. Accepted: 21 June 2021 Abstract Aim: The aim of this study was to evaluate the effectiveness of IFIO
in improving the safe relationship between couples among couples in
Dezful. Method: In this study, a single case experimental design of
multiple asynchronous baseline was used. The study population
included all couples who referred to psychological clinics in Dezful
in 1399. The research sample was selected using purposive sampling
method. The sample consisted of 4 couples (8 people) referred to
these clinics who were considered as insecure couples according to
the score of the Safe Communication Questionnaire (BARE). The
IFIO protocol was implemented in three phases: baseline, 8-session
intervention, and 3 to 1-month follow-up. Subjects answered the
BARE Questionnaire and the Intimate Security Questionnaire (ISQ)
before treatment, in the middle of treatment, at the end of treatment,
and every month after treatment for 3 months. Results: The results
of the data using visual drawing method, stable change index (RCI)
and recovery percentage formula showed that Intimacy From the
Inside Out therapy (IFIO) has a significant effect on increasing safe
communication and emotional security and couples' self-esteem. And
the couple were able to maintain the therapeutic effects in the
quarterly follow-up. Conclusion: Based on the data, it can be
concluded that (IFIO) has been effective in increasing the secure
relationship and Emotional Security, Secure to Being self of couples. Received: 3 May 2021 Key words: Inside Out Intimacy From the therapy (IFIO), Secure
Communication, Emotional Security, Secure to Being self Key words: Inside Out Intimacy From the therapy (IFIO), Secure
Communication, Emotional Security, Secure to Being self y words: Inside Out Intimacy From the therapy (IFIO), Secure
mmunication, Emotional Security, Secure to Being self Key words: Inside Out Intimacy From the therapy (IFIO), Secure
Communication, Emotional Security, Secure to Being self 297 مقاله پژوهشی
بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود
ارتباط ایمن زوجین
فصلنا
مة علمی-
پژوهشی
ن روا
شناسی کاربردی، دور
ة
15، شمار
ة 2 (
58
،)تابستان
1400
،
317
-
297
ISSN: 2008-4331- eISSN: 2645- 3541 نرگس حیدریان
فر1، خالد اصالنی2
*، غالمرضا رجبی3 و عباس
1
. دانشجوی دکتری مشاوره
،خانواده دانشگاه شهید چمران
،اهواز
،اهواز ایران . 2
. دانشیار گروه
،مشاوره دانشگاه شهید چمران
،اهواز
،اهواز ایران. :ایمیل
kh.aslani@scu.ac.ir
3
. استاد گروه
،مشاوره دانشگاه شهید چمران
،اهواز
،اهواز ن ایرا. چکیده :دریافت مقاله13
/
02
/
1400 :هدف
( بررسی اثربخشی درمان صمیمیت از درون به برونIFIO
) در بهبود ارتباط ایمن زوجین در
زوج
های مراجعه
کنند
ۀ
.شهرستان دزفول بود
روش :
در این پژوهش از طرح تجربی تک موردی از نوع خط
ۀ پای چندگان
ۀ ناهم
زمان استفاده شد. جامع
ۀ پژوهش شامل کلی
ۀ زوج
های مراجعه
کننده به کلینیک
های
روان شناختی شهرستان دزفول در سال1399
بود. نمون
ۀ
پژوهش شامل4
( زوج8
نفر) از افراد مراجعه
کننده این کلینیک
ها بودند که با توجه به نمر
ۀ پرسشنامه ارتباط ایمن (
BARE
) جزء زوج های با ارتباط
ناایمن قرار می گرفتند. پروتکل( درمان صمیمیت از درون به برونIFIO
) در سه مرحل
ۀ خط پایه، مداخل
ۀ
8 جلسه ای و3
پیگیری1 ماهه اجرا گردید و آزمودنی
ها قبل از درمان، میان
ۀ
درمان، پایان درمان و هر
ماه پس از پایان درمان به مدت3 ماه ب ه
پرسشنام
ۀ
( ارتباط ایمنBARE
) و
پرسشنام
ۀ
امنیت صمیمانه
(
ISQ
.) پاسخ دادند یافته
ها: نتایج حاصل از داده( ها به روش ترسیم دیداری، شاخص تغییر پایاRCI
)
و فرمول درصد بهبودی نشان داد که: درمان( صمیمیت از درون به برونIFIO
)
در افزایش ارتباط ایمن
و امنیت هیجانی و خود بودن زوج ها تأثیر
معنادار
داشته است و زوجین در پیگیری سه ماهه توانستند
اثرات درمانی را
حفظ
کنند. مقدم
ه ازدواج شامل پیوند دو نفر با عالیق، خواسته
ها و نیازهای مختلف است و رابط
ۀ ه ویژ ای است که
توسط قوانین و مقررات
اجتماعی صورت می
گیرد و به طور قابل توجهی بر رشد و تحقق
خود
افراد تأثیر می
گذارد. اگر چه ابعاد و مفاد ازدواج در طول تاریخ بشر تغییر کرد ه است، نهاد خانواده
و ازدواج جهانی بودن خود را حفظ کرده و هنوز نهاد اصلی جامعه هست (کوسیجیت اوز ،یجیت
2017
). اساس شکل گیری
و توسع
ۀ
.یک سیستم کوچک و اجتماعی روابط زن و شوهر است
روابط زوجی در زندگی افراد خیلی مهم است. این روابط می
تواند منبع شادی، و وقت ی که درست
نباشد منبع عمد
ۀ
،پریشانی و حتی بیماری باشد (پانزتی و ماچ2006
.)
از طرفی
رابط
ۀ
بین
،فردی نوعی وابستگی یا آشنایی
،قوی عمیق، یا نزدیک بین دو یا چند نفر است که می
تواند
کوتاه یا بلندمدت باشد؛ این وابستگی ممکن است بر ۀ پای
،استنتاج، عشق، همبستگی
ت عامل
،تجاری منظم، یا برخی از انواع دیگر تعهد اجتماعی باشد (آرنولد و بوگز2015
.) ازدواج شامل پیوند دو نفر با عالیق، خواسته
ها و نیازهای مختلف است و رابط
ۀ ه ویژ ای است که
توسط قوانین و مقررات
اجتماعی صورت می
گیرد و به طور قابل توجهی بر رشد و تحقق
خود
افراد تأثیر می
گذارد. اگر چه ابعاد و مفاد ازدواج در طول تاریخ بشر تغییر کرد ه است، نهاد خانواده
و ازدواج جهانی بودن خود را حفظ کرده و هنوز نهاد اصلی جامعه هست (کوسیجیت اوز ،یجیت
2017
). اساس شکل گیری
و توسع
ۀ
.یک سیستم کوچک و اجتماعی روابط زن و شوهر است
روابط زوجی در زندگی افراد خیلی مهم است. مقدم
ه این روابط می
تواند منبع شادی، و وقت ی که درست
نباشد منبع عمد
ۀ
،پریشانی و حتی بیماری باشد (پانزتی و ماچ2006
.)
از طرفی
رابط
ۀ
بین
،فردی نوعی وابستگی یا آشنایی
،قوی عمیق، یا نزدیک بین دو یا چند نفر است که می
تواند
کوتاه یا بلندمدت باشد؛ این وابستگی ممکن است بر ۀ پای
،استنتاج، عشق، همبستگی
ت عامل
،تجاری منظم، یا برخی از انواع دیگر تعهد اجتماعی باشد (آرنولد و بوگز2015
.)
پژوهش
ها نشان می
دهند که رفتارهای قابل سنجشی در رو ابط میان زن و مرد وجود دارند که
با رضایت بیشتر از رابطه و دلبستگی1 ایمن ،تر مرتبط هستند (ساندبرگ، بازبی، جانسون و یوشیدا
2012) و این
که رفتارهای دلبستگی ایمن می توانند به عنوان حایلی در برابر نامالیمات موجود
،در رابطه عمل کنند (کناپ، ساندبرگ، نواک و الرسون
2015
،؛ اوکا، ساندبرگ، بارادفورد و براون
2014). از طرفی عدم ایمنی در دلبستگی با نگرانی های مربوط به بهداشت فکری و فیزیکی از
)جمله (عالئم افسردگی، خشونت، شدت درد و بهبود عملکردی از خستگی مزمن و فیبرومیالژی
،مرتبط است (هامیل2010
،، مکوی2005؛ اوکا، ساندبرگ و
،اوکا2014
،؛ ریس و گرنیر2004
؛
،تریمبالی و سالیوان2010
). ما باید نقش اساسی دلبستگی را در سالمت رابطه درک کنیم
،(سدال و وامپلر2013). پژوهش
های صورت گرفته بر روی زوجین و دلبستگی میان آن ها نشان
می دهد که رفتارهای حاکی از پشتیبانی باعث دلبستگی و نیز رضایت
از رابطه می ،شود (فینی
2002
،؛ رولس، کمپبل سیمسون و گریچ2001). با این وجود در مورد سبک های دلبستگی
،عمومی در مقایسه با رفتارهای دلبستگی خاص اطالعات خیلی بیشتری وجود دارد (آینسورث
،بلرهار، واترز و وال1978
،؛ بالبی1969
،؛ فینی2008
( ). بنابراین بالبی1973) قا بلیت دسترسی
و پاسخگویی را در اصل به عنوان رفتارهای کلیدی شرح داد که امنیت یا درماندگی در روابط
نزدیک را تعیین می( کنند. 1 - Attachment Key words: Inside Out Intimacy From the therapy (IFIO), Secure
Communication, Emotional Security, Secure to Being self نتیجه
گیری
: براساس داده
ها می توان نتیجه گرفت که درمان صمیمیت از
( درون به برونIFIO
)
در افزایش ارتباط ایمن و امنیت
هیجانی و خود بودن زوج
ها
مؤثر بوده است و
ارتباط ایمن زوج
ها را ارتقا می.دهد :پذیرش مقاله31
/
03
/
1400 298
:استناد به این مقاله
حیدریان
فر، نرگس.، اصالنی، خالد.، رجبی، غالمرضا.، و امان ،الهی
عباس( . 1400
.)بررسی اثربخشی درمان صمیمیت
از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین .فصلنامۀ علمی- پژوهشی روان،شناسی کاربردی
15
(2
پیاپی58
:)
317
-
297
. 298 فصلنام
ة علمی-
پژوهشی
ن روا
شناسی
،کاربردی ة دور
15
، شمار
ة 2 (
58
)، تابستان
1400
،
317
-
297
مقدم
ه بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین پیوند دهنده در پی آن به وجود می آید؛ که این لحظه"پذیرندگی1" نامیده می
شود. پژوهش های
به عمل آمده بر روی ارتباط ایمن زوجین نشان می
دهند که رفتارهای نشان دهند
ۀ
،دسترسی
،پاسخگویی و پذیرندگی با دلبستگی ایمن مرتبط است (ساندبرگ، بازبی، جانسون و یوشیدا
2012
.) انجام ده ها سال پژوهش و بررسی نشان داده است که وجود یک شخص و ایجاد حس
دلبستگی در بزرگسالی می تواند باعث باال رفتن حس امنیت، راحتی و تأثیر مثبت شود
،(میکولینسر و شاور2007). همچنین در ارتباط با روابط عاشقان
ۀ پژوهش
ها نشان می دهد که
زوج
های دارای دلبستگی با ،التر دارای کیفیت بهتری در برقراری ارتباط هستند (فینی2008
.)
بنابراین روابط صمیمانه هست
ۀ ارتباطات اجتماعی ما را تشکیل می
دهند، و به ما کمک می کنند
رشد کنیم، عمیقاً عشق بورزیم و ناکامی ها را تحمل کنیم (بالبی1969
،؛ کوان2006
،؛ جانسون
2004
،؛ جانسون و گریمن2013). وقتی که زوجین وارد درمان می ،شوند، در وضعیت قطع ارتباط
خشم و اندوه و از دست دادن صمیمیت به سر می برند، و اغلب دو هدف را از ما درخواست
می .کنند: احساس امنیت و بازسازی صمیمیت از دست رفته
درمان صمیمیت از درون به بیرون2
(
IFIO
)
نقش
ۀ راهی را برای رسیدن ب
ه این هدف
ها ارائه می دهد
(هربین-
بالنک، کرپلمن و
،اسویزی2016
.)
( درمان صمیمیت از درون به بیرونIFIO) یک مدل تجربی از درمان زوج ها
است که حدود ده سال پیش از مفاهیم سیستم
های خانواده درونی3 (
IFS
،) برای روابط صمیمی
و وسیله ای برای رشد و بهبود روابط زوجین به وجود آمد. این رویکرد به زبان واضح و روشن بیان
می کند که چگونه ممکن است دو سیستم داخلی یک زوج با یکدیگر برخورد
کنند و آن ها را به
سمت فروپاشی رابطه ببرد. ممکن است بخش
های یک زوج در واکنش های افراطی داشته باشند
و یا عقاید تند و افراطی را از سال
های دور با خ ود آورده باشند، در چنین شرایطی اگر زوجین
احساس امنیت بکنند، می توانند این بار سنگین را رها کرده و واکنش دیگری را برگزینند. این
مدل زوج درمانی به دنبال یافتن نقاط قوت و منابع هر شریک، به منظور التیام زخم هایی است
که از روابط قبلی بر جای مانده
اند یا در کودک
ی رخ داده اند، و هنوز وجود دارند. هدف پرورش
سالم، ارتباط صمیمی بین زوجین است. 1 - Acceptance
2 - Intimacy From the Inside Out
therapy مقدم
ه جانسون2008
) به این نتیجه رسید که زمانی که زوجین در هنگام
نیاز در دسترس یکدیگر باشند و به صورت آرام کننده به نیازهای هم پاسخ دهند، یک لح
ظۀ ،تجاری منظم، یا برخی از انواع دیگر تعهد اجتماعی باشد (آرنولد و بوگز2015
.)
پژوهش
ها نشان می
دهند که رفتارهای قابل سنجشی در رو ابط میان زن و مرد وجود دارند که
با رضایت بیشتر از رابطه و دلبستگی1 ایمن ،تر مرتبط هستند (ساندبرگ، بازبی، جانسون و یوشیدا
2012) و این
که رفتارهای دلبستگی ایمن می توانند به عنوان حایلی در برابر نامالیمات موجود
،در رابطه عمل کنند (کناپ، ساندبرگ، نواک و الرسون
2015
،؛ اوکا، ساندبرگ، بارادفورد و براون
2014). از طرفی عدم ایمنی در دلبستگی با نگرانی های مربوط به بهداشت فکری و فیزیکی از
)جمله (عالئم افسردگی، خشونت، شدت درد و بهبود عملکردی از خستگی مزمن و فیبرومیالژی
،مرتبط است (هامیل2010
،، مکوی2005؛ اوکا، ساندبرگ و
،اوکا2014
،؛ ریس و گرنیر2004
؛
،تریمبالی و سالیوان2010
). ما باید نقش اساسی دلبستگی را در سالمت رابطه درک کنیم
،(سدال و وامپلر2013). پژوهش
های صورت گرفته بر روی زوجین و دلبستگی میان آن ها نشان
می دهد که رفتارهای حاکی از پشتیبانی باعث دلبستگی و نیز رضایت
از رابطه می ،شود (فینی
2002
،؛ رولس، کمپبل سیمسون و گریچ2001). با این وجود در مورد سبک های دلبستگی
،عمومی در مقایسه با رفتارهای دلبستگی خاص اطالعات خیلی بیشتری وجود دارد (آینسورث
،بلرهار، واترز و وال1978
،؛ بالبی1969
،؛ فینی2008
( ). بنابراین بالبی1973) قا بلیت دسترسی
و پاسخگویی را در اصل به عنوان رفتارهای کلیدی شرح داد که امنیت یا درماندگی در روابط
نزدیک را تعیین می( کنند. جانسون2008
) به این نتیجه رسید که زمانی که زوجین در هنگام
نیاز در دسترس یکدیگر باشند و به صورت آرام کننده به نیازهای هم پاسخ دهند، یک لح
ظۀ 1 - Attachment 299 3 - Internal Family Systems بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین اما انجام پژوهش
هایی مورد نیاز است که
رفتارهایی را
بررسی
کند، که منجر به دلبستگی ایمن در روابط زوجین می شود. درمانIFIO
یک رویک رد
جدید است که در کار با زوجین پژوهش
های کمی انجام شده است و در داخل کشور درما ن
مشابهی که بتوان ارتباط این رویکرد درمانی را در کار با زوجین نشان دهد وجود ن
دارد ، و حتی
در خارج از کشور نیز
کار با زوجین به صورت پژوهش انجام نشده است. بنابراین با
ت وجه به خالء
:پژوهش مرتبط، پژوهش حاضر درصدد پاسخگویی به این سؤاالت است
1. آیا درمان
صمیمیت از درون به برون در بهبود ارتباط ایمن زوج
ها موثر است؟
2. آیا درمان
صمیمیت از درون به برون در بهبود امنیت هیجانی زوج
ها مؤثر است؟
3. آیا درمان
صمیمیت از درون به برون در بهبود امنیت خود بودن زوج
ها مؤثر است؟ بخش هایمان یک"خود1" قدرتمند در درون داریم. به نظر می رسد که"خود"
با صدایی مهربان
و دلسوز و با توانایی برای ترمیم دنیای درونی ظاهر می
شود و می تواند رهبری سیستم را به دست
گیرد. هر چه"انرژی خود" زوجین بیشتر باشد در نظر افراد دیگر قابل اعتمادتر به نظر می .رسد
چیزی که هدف درمان محسوب می.شود
در درمانIFIO
به جای این که به زوجین کمک کنیم
تا مشکل را حل کنند یا راه
حلی برای تعارضات اجتناب ناپذیر خود پیدا کنند، همسران را به سمت
رضایت متقابل و هر چیزی که رابط
ۀ آن
ها را با معنی کند، هدایت می
کنیم. درمان قصد دارد
پیوند
ی امن را پرورش ده
د که تقویت کنن
دۀ تعامالت باز، مراقبت از شریک و فهم تفاوت ها و نیز
شباهت
ها
ی همدیگر است. وقتی که زوج به انداز
ۀ
کافی احساس امنیت کنند تا
همزمان بخش های
خود و شریکشان
را درک کنند
، تعامالت آن
ها به تغییرات شدیدی گرایش پیدا می
کند. احساس
امنیت موجب می
شود تا زوجین از اضطراب و تنش کم تری در روابط خود برخوردار باشند و با
احساس سرزندگی و شادکامی به زندگی زناشویی خود بپ ردازند و بتوانند به خوبی روابط خود را
با همسرشان مدیریت کنند و در نهایت به رابطه
ای صمیمانه دست پیدا کنند
،(هاگ، جانسن
،کریستینسن، فریدریکسن، دالتون، دیرگرو و همکاران2018
). بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین هدایت دو نفر و بخش
های آن ها برای تعامل با یکدیگر
و آغوش باز است (هربین-
،بالنک و همکاران2016
.) بازسازی رابط
ۀ عاشقانه می
تواند زخم های
کهنه را التیام بخشد. در طول درمان درمانگر
با چیزی متفاوت از بخش های مراجع ارتباط برقرار
می کند: چیزی که از آن به عنوان"خود من"
یاد می کند. به این صورت که ما عالوه بر
1 - Acceptance
2
3 - Internal Family Systems 300 صلنام
ة علمی-
پژوهشی
ن روا
شناسی
،کاربردی ة دور
15
، شمار
ة 2 (
58
)، تابستان
1400
،
317
-
297 بخش هایمان یک"خود1" قدرتمند در درون داریم. به نظر می رسد که"خود"
با صدایی مهربان
و دلسوز و با توانایی برای ترمیم دنیای درونی ظاهر می
شود و می تواند رهبری سیستم را به دست
گیرد. هر چه"انرژی خود" زوجین بیشتر باشد در نظر افراد دیگر قابل اعتمادتر به نظر می .رسد
چیزی که هدف درمان محسوب می.شود
در درمانIFIO
به جای این که به زوجین کمک کنیم
تا مشکل را حل کنند یا راه
حلی برای تعارضات اجتناب ناپذیر خود پیدا کنند، همسران را به سمت
رضایت متقابل و هر چیزی که رابط
ۀ آن
ها را با معنی کند، هدایت می
کنیم. درمان قصد دارد
پیوند
ی امن را پرورش ده
د که تقویت کنن
دۀ تعامالت باز، مراقبت از شریک و فهم تفاوت ها و نیز
شباهت
ها
ی همدیگر است. وقتی که زوج به انداز
ۀ
کافی احساس امنیت کنند تا
همزمان بخش های
خود و شریکشان
را درک کنند
، تعامالت آن
ها به تغییرات شدیدی گرایش پیدا می
کند. احساس
امنیت موجب می
شود تا زوجین از اضطراب و تنش کم تری در روابط خود برخوردار باشند و با
احساس سرزندگی و شادکامی به زندگی زناشویی خود بپ ردازند و بتوانند به خوبی روابط خود را
با همسرشان مدیریت کنند و در نهایت به رابطه
ای صمیمانه دست پیدا کنند
،(هاگ، جانسن
،کریستینسن، فریدریکسن، دالتون، دیرگرو و همکاران2018
). اگر چه پژوهش های گذشته حاکی از این است که امنیت در رابطه، با میزان ارتباط
ز ،وجین
ع
صبانیت و خشونت در روابط، استفاده از بازی
های ویدیویی، مشکالت مربوط به خانو
اده های
اصلی، خود گردانی، عزت نفس، میزان فعالیت فیزیکی و عادات غذایی مرتبط است (کنا پ و
،همکاران2015
،؛ اوکا و همکاران2014). بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین ر ن
ر رو و
ون
ن
ن ر ر
ر
اگر چه پژوهش های گذشته حاکی از این است که امنیت در رابطه، با میزان ارتباط
ز ،وجین
ع
صبانیت و خشونت در روابط، استفاده از بازی
های ویدیویی، مشکالت مربوط به خانو
اده های
اصلی، خود گردانی، عزت نفس، میزان فعالیت فیزیکی و عادات غذایی مرتبط است (کنا پ و
،همکاران2015
،؛ اوکا و همکاران2014). اما انجام پژوهش
هایی مورد نیاز است که
رفتارهایی را
بررسی
کند، که منجر به دلبستگی ایمن در روابط زوجین می شود. درمانIFIO
یک رویک رد
جدید است که در کار با زوجین پژوهش
های کمی انجام شده است و در داخل کشور درما ن
مشابهی که بتوان ارتباط این رویکرد درمانی را در کار با زوجین نشان دهد وجود ن
دارد ، و حتی
در خارج از کشور نیز
کار با زوجین به صورت پژوهش انجام نشده است. بنابراین با
ت وجه به خالء
:پژوهش مرتبط، پژوهش حاضر درصدد پاسخگویی به این سؤاالت است
1. آیا درمان
صمیمیت از درون به برون در بهبود ارتباط ایمن زوج
ها موثر است؟
2. آیا درمان
صمیمیت از درون به برون در بهبود امنیت هیجانی زوج
ها مؤثر است؟
3. آیا درمان
صمیمیت از درون به برون در بهبود امنیت خود بودن زوج
ها مؤثر است؟ 1 - Self 301 301 بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین روش
در این پژوهش از طرح آزمایشی تک
موردی استفاده شده است. و از بین طرح های آزمایشی
تک موردی از طرح تجربی
قبل-
بعدA-B
با پی
گیری انتخاب شده است (طرح خط پایه ی
چندگا
نه1). 1 - Multiple baseline 2 - Baseline Phases 3 - The Brief Accessibility,
Responsiveness, and Engagement
(BARE) Scale بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین در پژوهش حاضر آزمودنی
ها به صورت همزمان وارد مرحل
ۀ خط پایه2
و با قرار گرفتن
به صورت تصادفی در یکی از دوره
های انتظار دو ،سه یا چهار هفته
ای در مرحل
ۀ
خط پایه وارد
مرحل
ۀ
درمان8 جلسه ای و سپس پیگیری (در طی
سه دوره ی1
)ماهه.شدند جامع
ۀ
آماری این
پژوهش
ۀ کلی زوج
های مراجعه
کننده به کلینیک
های روان شناختی دزفول در سال1399
.بود
نمون
ۀ پژوهش با استفاده از روش نمونه گیری هدفمند انتخاب شدند، نمونه شامل4
( زوج8
)نفر
از افراد مراجعه کننده به این کلینیک
ها بود که با توجه به نمر
ۀ پرسشنام
ۀ
( ارتباط ایمنBARE
)
جزء زوج
های با ارتباط ناایمن قرار می.گرفتند روش نمونه
گیری در این پژوهش نمونه گیری
هدفمند بود به این صورت که زوج
ها پس احراز شرایط ورود و پر کردن پرسش
نامه
ها وارد دور
ۀ
درمانی می.شوند ابزارهای پژوهش
1. پرسشنام
ة ارتباط ایمن3 (
BARE
):
مقیاس ارتباط ایمن توسط ساندبرگ، بازبی، جانسون
( و یوشیدا2012
) ساخته شد. یک ابزار12
آیتمی است که رفتارهای کلیدی را در سیستم
دلبستگی زوج ارزیابی می( کند. بالبی1973
) قابلیت دسترسی و پاسخگویی را در اصل به عنوان
رفتارهای کلیدی شرح می
دهد که امنیت یا درماندگی در روابط نزدیک را تعیین می کند. در این
مقیاس آیتم
های موجود در بخش های«قابلیت دسترسی و دلبستگی »
به صورت معکوس
نمره
گذاری می
شود (سؤاالت شماره 1
،
2
،
5
،
6
،
7
،
8
،
11
،
12). نمره گذاری به این صورت است
که آیتم
های موجود در هر دسته
بندی فرعی جمع می
شود و یا بر ای کل مقیاس دسته بندی
فرعی نمرات با هم جمع می
شود، امتیاز باالتر به معنای رفتارهای گزارش شدۀ وابست
ۀ
بیشتر
است. در این مقیاس نمرات از طریق امتیاز دهی به خود و امتیاز به همسر (که در آن شما همسر
خود را درجه بندی می
کنید) به دست می( آیند. ساندبرگ2012) دستورال
عمل های زیر را از
مطالعه در نمونه )هایی که از نظر جمعیت شناسی کلیدی شبیه گروه نمونه ملی (دین، نژاد و غیره
است به دست آوردند. بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین آن ها امتیازها را به سه رنگ قرمز، زرد و سبز تقسیم کردند که در این 302 صلنام
ة علمی-
پژوهشی
ن روا
شناسی
،کاربردی ة دور
15
، شمار
ة 2 (
58
)، تابستان
1400
،
317
-
297 تقسیم بندی قرمز به معنای نیاز به قدری کار، زرد به معنای قدری توجه
و سبز به معنای خوب
است. به ویژه تقسیم
بندی پایین، متوسط، باال نشان دهنده ی امتیاز دهیBARE
و ارتباط آن ها
با امتیازهای مربوط به کیفیت زناشویی (محیط های مشکل دار، رضایت، ثبات)؛ قرمز به معنای
.کیفیت زناشویی پایین، زرد به معنای متوسط و سبز به معنای باال است اگر چه چند دسته بندی
فرعی برای زنان و مردان از نظر آماری متفاوت است ولی این تفاوت ها از نظر بالینی مهم و یا
بامعنا نبودند. آن
ها استفاده از جمع کل امتیازها را توصیه می کنند چون ممکن است محدودۀ
موجود در دسته بندی
های فرعی تکی، گستردگی کافی جهت کمک به پزشک ب الینی و یا شخص
مراجعه کننده را نداشته باشد. در این پژوهش ضریب پایایی با استفاده از روش آلفای کرانباخ
92
/0
.به دست آمد 2. پرسشنام
ة امنیت صمیمانه1 (
ISQ
:)پرسشنام
ۀ
امنیت صمیمانه توسط کوردووا، جی و
( وارن2005
) تدوین شده و دارای28
سوال است و به منظور ارزیابی سطح آرامش افراد از
آسیب
پذیر بودن در کنار شریک خود و در بافت حوزه
های مختلف رابط
ۀ
طراحی شده است. این
پرسشنامه دارای5
،زیر مقیاس امنیت هیجانی، امنیت جسمانی/ جنسی
امنیت خود بودن، امنیت
در عموم و امنیت مخالفت و دارای امتیاز کلی0
تا112
است. سؤاالت این مقیاس براساس5
امتیازی لیکرت از0
(هرگز) تا4 (همیشه) درجه بندی می شوند. هر چه امتیاز فرد بیشتر باشد
وی امنیت صمیمانه بیشتری را تجربه می
کند. سؤال های3
،
5
،
7
،
8
،
11
،
14
،
15
،
16
،
20
،
23
،
24
،
26
،
27
و28
به صورت معکوس نمره
گذاری می( شوند. در مطالعه دآن هام2008
؛ به نقل
( از بارتوس2017
) همسانی درونی این پرسشنامه برابر با88
/0 بدست آمد. همچنین در مطالعه ای
دیگر،
او آلفای93
/0
را برای مردان و96
/0 را برای زنان و اعتبار با زآزمایی در طول دوره یک
ماهه را برای مردان83
/0
و برای زنان92
/0
،به دست آورد (بارتوس2017
). در مطالعه کوردورا
( و همکاران2005
) همسانی درونی این مقیاس باال (آلفای90
/0) گزارش شد. 303
1 - The Intimate Safety Questionnaire
(ISQ)
2 -Personal Assessment of Intimacy
in Relationships Questionnaire
3 - Schaefer, & Olson
4 - Intellectual Intimacy
5 - Emotional Intimacy
6 - Snyder 2 -Personal Assessment of Intimacy
in Relationships Questionnaire 1 - The Intimate Safety Questionnaire
(ISQ)
2 بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین در مطالع
ۀ آن ها
ارتباط معناداری بین این مقیاس با پرسشنام
ۀ
ارزیابی شخصی صمیمیت
در روابط2 (
PAIR
؛
اسچیفر و اولسون3 ،
1981) یافت شد. به ویژه در خرده مقیاس صمیمیت فکری4 (
78
rs=
برای
زنان و73
rs=
برای مردان) و خرده مقیاس صمیمیت عاطفی5 (
82
rs=
برای زنان و80
rs=
برای مردان). این نشان می دهد کهISQ
وPAIR
ساختارهای بسیار مشابهی را اندازه گیری
می کنند. عالوه بر اینISQ
با انداز
ۀ پریشانی کلی در مقیاس رضایت زناشویی (اسنایدر6 ،
1979
؛ 303
1 - The Intimate Safety Questionnaire
(ISQ)
2 -Personal Assessment of Intimacy
in Relationships Questionnaire
3 - Schaefer, & Olson
4 - Intellectual Intimacy
5 - Emotional Intimacy
6 - Snyder 3 - Schaefer, & Olson 4 - Intellectual Intimacy 2 -Personal Assessment of Intimacy
in Relationships Questionnaire 1 -Marital Status Inventory
2 - Weiss, & Cerreto بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین 72
rs=
برای زنان و68
rs=
برای مردان)، مقیاس وضعیت ازدواج1 ( ویس و کِرتو2 ،
1980
؛
54
rs=
برای زنان و43
rs=
برای مردان) و سبک
های دلبستگی زوجین3 (هازان و شاور4 ،
1990
؛
rs=42
برای زنان وrs=43
برای مردان) ارتباط معناداری دارد. این نتایج حمایت اولیه برای
ِساخت و معیار اعتبارISQ
فراهم می کنند. در پژوهش حاضر از سؤاالت مربوط به خرده مقیاس
امنیت هیجانی و امنیت خود بودن استفاده شده است. در پژوهش حاضر تیز ضریب پایایی با
استفاده از رو ش آلفای کرانباخ برای مقیاس کل96
/0
و برای خرده مقیاس امنیت هیجانی89
/0
و برای امنیت خود بودن90
/0
.به دست آمد
3. برنام
ة
:درمانی صمیمیت از درون به برون
منظور از مداخل
ۀ درمانی صمیمیت از د رون به
برون، مداخله
ای است که براساس کتاب درمانی هربین-ب النک و همکاران(
2016
) برای کار با
زوجین به کار برده است. این مدل یک درمان اقتباسی است که از نظریه سیستم
های خا نواده
( درونیIFS) در کار با زوجین به کار گرفته شده است. و شامل یک دور
ۀ درمان سه مر
حله ای
در8
:جلسه درمانی به شرح زیر است 72
rs=
برای زنان و68
rs=
برای مردان)، مقیاس وضعیت ازدواج1 ( ویس و کِرتو2 ،
1980
؛
54
rs=
برای زنان و43
rs=
برای مردان) و سبک
های دلبستگی زوجین3 (هازان و شاور4 ،
1990
؛
rs=42
برای زنان وrs=43
برای مردان) ارتباط معناداری دارد. این نتایج حمایت اولیه برای
ِساخت و معیار اعتبارISQ
فراهم می کنند. در پژوهش حاضر از سؤاالت مربوط به خرده مقیاس
امنیت هیجانی و امنیت خود بودن استفاده شده است. در پژوهش حاضر تیز ضریب پایایی با
استفاده از رو ش آلفای کرانباخ برای مقیاس کل96
/0
و برای خرده مقیاس امنیت هیجانی89
/0
و برای امنیت خود بودن90
/0
.به دست آمد 3. برنام
ة
:درمانی صمیمیت از درون به برون
منظور از مداخل
ۀ درمانی صمیمیت از د رون به
برون، مداخله
ای است که براساس کتاب درمانی هربین-ب النک و همکاران(
2016
) برای کار با
زوجین به کار برده است. این مدل یک درمان اقتباسی است که از نظریه سیستم
های خا نواده
( درونیIFS) در کار با زوجین به کار گرفته شده است. و شامل یک دور
ۀ درمان سه مر
حله ای
در8
:جلسه درمانی به شرح زیر است جدول1. 3 - partners’ attachment styles
4 - Hazan, & Shaver بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین پروتکل استفاده ازIFIO
در بهبود ارتباط ایمن زوجین (به نقل از هربین-
بالنک و
همکاران ،
2016
) مرحله1: شروع
مالقات با زوجین برای ارزیابی سطح تمایز آن.ها
بررسی این
که از چه چیزی می.ترسند
بررسی این که چه چیزی می.خواهند؛ امیدها، تمایالت و اهداف
فراهم کردن امکانات
مرحله2
: جریان و
چرخه
ی میانه درمان
آموختن مهارت.های ارتباطی جدید به زوجین
کسب مجوز از مراجع تا نقش ردیاب بخش.ها را به عهده بگیرید
آسیب
پذیری و شیوه
ی دعوا کردن آن:ها را پیگیری کنید. شامل
نحوه مذاکره در مورد نیازها
چگونگی تجربه شرم
ارتقا و تقویت التیام رابطه
ای
انجام ک:ارهای فردی شامل
عدم آمیختگی 304 فصلنام
ة علمی-
پژوهشی
ن روا
شناسی
،کاربردی ة دور
15
، شمار
ة 2 (
58
)، تابستان
1400
،
317
-
297
آمیخته نشدن با شرم
مرحله سوم: پایان
پیشبرد روند ترمیم
پرداختن به بخشش
و در صورت پیشروی، ایجاد رابطه مبتنی بر تأیید تفاوت
ها، و به اشتراک گذاری دیدگاه.ها ة شیو اجرا هر
چهار
زوج در مجموع8 جلس
ۀ
.درمانی دریافت کردند 305 305 بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین یافته
ها
در این پژوهش4
( زوج8
.نفر) به عنوان آزمودنی شرکت کردند :که زوج اول در ردۀ سنی (مرد
34
:و زن29
)، دارای تحصیالت (مرد: دیپلم، زن: لیسانس) و مدت زمان ازدواج8
.سال بودند
:زوج دوم در ردۀ سنی (مرد30
:و زن27
)، دارای تحصیالت (مرد: دیپلم، زن: لیسانس) و مدت
زمان ازدواج5
:سال بودند. زوج سوم در ردۀ سنی (مرد39
:و زن34
:)، دارای تحصیالت (مرد
لیسانس، زن: دیپلم) و مدت زمان ازدواج3
:سال بودند. و زوج چهارم در ردۀ سنی (مرد29
و
:زن25
)، دارای تحصیالت (مرد: دیپلم، زن: دیپلم) و مدت زمان ازدواج2
سال بودند. در مجموع
از تعد اد8
آزمودنی5
( نفر دیپلم5/
%62
) و3
(نفر لیسانس5/
37%) بودند. و آزمودنی ها دارای
( میانگین سنی30.75
( ) و4.5
.) سال مدت ازدواج بودند
جدول2) نمره
های چهار زوج در پرسش
ۀ نام ارتباط ایمن
را بر حسب مراحل مختلف درمان را
نشان می.دهد ة شیو اجرا پس از این که افراد واجد شرایط انتخاب شدند، ۀ هم آزمودنی
ها به طور همزمان وارد مرحل
ۀ
خط
پایه شدند. برای کنترل متغیرهای ناخواسته و مداخله
گر احتمالی طبق اصول طرح
های خط پای
ۀ
چندگانه زوج
ها به طور تصادفی از نظر ترتیب ورود یک به یک با فاصل
ۀ
)یک جلسه (یک هفته
وارد طرح درمان شدند. به این نحو که زوج اول پس از دو جلسه سنجش در مرحل
ۀ
خط پایه
یعنی از هفت
ۀ سوم وارد جلس
ۀ اول درمان، و بعد از آن زوج دوم در جلس
ۀ دوم زوج اول وارد جلس
ۀ
اول درمان و زوج سوم نیز در جلس
ۀ دوم زوج دوم و جلس
ۀ سوم زوج اول، وارد ج
ۀ لس
اول و به
همین ترتیب زوج چهارم در جلسۀ چهارم زوج اول و جلسۀ سوم زوج دوم و جلسۀ دوم زوج سوم
وارد طرح درمان شدند
. همچنین ضمن اجرای درمان، کلی
ۀ پرسش نامه
ها در جلسات 3، 5
،
8
و
پس از درمان در فاصل
ۀ زمانی یک ماه
)به یک ماه (سه تا سنجش یک ماهه مرحل
ۀ پیگیری ان جام
.شد الزم به ذکر می باشد طرح
درمان صمیمیت از درون به برون، مداخله ای است که براساس کتاب
درمانی هربین-
( بالنک و همکاران2016
) برای کار با زوجین به کار برده است. این مدل یک
درمان اقتباسی است که از نظریه سیستم( های خانواده درونیIFS) در کار با زوجین به کار
گرفته شده است. و شامل یک دوره درمان سه مرحله ای در8
.جلسه درمانی است
طرح درمان
شامل
مرحله شروع (مالقات با زوجین برای ارزیابی سطح تمایز آن
ها، فراهم کردن امکانات ،)
میانۀ درمان (آموختن مهارت های ارتباطی جدید به زوجین،کسب مجوز از مراجع برای نقش
ردیاب بخش
ها
، بررسی آسیب
پذیری و شیوۀ دعوا کردن آن
ها ،بردن آن ها به درون و عمیق کردن
کار با تمرینات تجربی ،ارتقا و تقویت التیام رابطه
ای) و مرحله پایان (پیشبرد روند ترمیم ،
پرداختن
به بخشش ) است. زوج اول یافته
ها در این پژوهش4
( زوج8
.نفر) به عنوان آزمودنی شرکت کردند :که زوج اول در ردۀ سنی (مرد
34
:و زن29
)، دارای تحصیالت (مرد: دیپلم، زن: لیسانس) و مدت زمان ازدواج8
.سال بودند
:زوج دوم در ردۀ سنی (مرد30
:و زن27
)، دارای تحصیالت (مرد: دیپلم، زن: لیسانس) و مدت
زمان ازدواج5
:سال بودند. زوج سوم در ردۀ سنی (مرد39
:و زن34
:)، دارای تحصیالت (مرد
لیسانس، زن: دیپلم) و مدت زمان ازدواج3
:سال بودند. و زوج چهارم در ردۀ سنی (مرد29
و
:زن25
)، دارای تحصیالت (مرد: دیپلم، زن: دیپلم) و مدت زمان ازدواج2
سال بودند. در مجموع
از تعد اد8
آزمودنی5
( نفر دیپلم5/
%62
) و3
(نفر لیسانس5/
37%) بودند. و آزمودنی ها دارای
( میانگین سنی30.75
( ) و4.5
.) سال مدت ازدواج بودند جدول2. روند تغییر مراحل درمان چهار زوج در
پرسش
نامه
ی ارتباط ایمن
گروه
های درمانی درمان صمیمیت از درون به برون
آزمودنی
ها
مراحل درمان
زوج اول زوج دوم زوج سوم زوج چهارم
میانگین مراحل خط پایه
10
66
/
14
5/
12
4/
12
میانگین مراحل درمان
33
/
17
20
33
/
20
33
/
22
شاخص)تغییر پایا (درمان
19
/8
94
/4
69
/7
87
/8
درصد بهبودی پس از درمان
3/
73
42
/
36
64
/
62
08
/
80
درصد بهبودی کلی پس از درمان
11
/
63
پیگیری یک ماهه
پیگیری دو ماهه
پیگیری سه ماهه
)شاخص تغییر پایا (درمان
درصد بهبودی پس از درمان
21
20
20
28
/
14
100
24
24
24
49
/5
71
/
63
24
25
25
53
/6
100
20
20
20
07
/4
29
/
61
درصد بهبودی کلی پس از درمان
25
/
81 306 زوج اول نمودار1. روند تغییر نمره
های پرسش
ۀ نام ارتباط ایمن زوج
ها در مراحل خط پایه، مداخله و پ
0
10
20
30
زوج اول
0
10
20
30
خط پایه1
خط پایه2
خط پایه3
جلسه3
جلسه5
جلسه8
پیگیری1
پیگیری2
پیگیری3
زوج دوم
0
10
20
30
خط پایه1
خط پایه2
خط پایه3
خط پایه4
جلسه3
جلسه5
جلسه8
پیگیری1
پیگیری2
پیگیری3
زوج
سوم
0
10
20
30
خط پایه1
خط پایه2
خط پایه3
خط پایه4
خط پایه5
جلسه3
جلسه5
جلسه8
پیگیری1
پیگیری2
پیگیری3
زوج چهارم بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین جدول2) نشان می دهد که مقدار شاخص تغییر پایا در زوج اول، دوم و سوم و چهارم در پایان
درمان به ترتیب19
/8
،
94
/4
،
69
/7
و87
/8
به دست آمد که با توجه به( سطح معناداری05
/0
p>
) تمامی نمرات به باالتر از1.96
z=
رسیده است و بنابراین می توان گفت تغییرات هر چهار
زوج نظر آماری قابل قبول و
نتیجه ی تأثیرات درمانی است. همین طور مقدار شاخص تغییر پایا
در زوج اول، دوم و سوم و چهارم در جلس
ۀ
پیگیری به ترتیب28
/
14
،
49
/5
،
53
/6
و07
/4
به
دست آمد که با توجه به( سطح معناداری05
/0
p>) نمر
ۀ
هر چهار زوج به باالتر از1.96
z=
.رسیده است
که این میزان بهبودی در هر چهار زوج بیانگر موفقیت در درمان می .باشد بنابراین
می
توان نتیجه گرفت درمان صمیمیت از درون به برون در افزایش ارتباط ایمن زوج ها مؤثر بوده
.است جدول3) نمره های چهار زوج در خرده مقیاس امنیت هیجانی را بر حسب مراحل مختلف درمان
را نشان می.دهد جدول3. زوج اول بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین روند تغییر مراحل درمان چهار زوج در خرده مقیاس
امنیت هیجانی
گروه
های درمانی درمان صمیمیت از درون به برون
آزمودنی
ها
مراحل درمان
زوج اول زوج دوم زوج سوم زوج چهارم
میانگین مراحل خط پایه
12
66
/
16
18
6/
16
میانگین مراحل درمان
66
/
18
66
/
28
66
/
30
28
شاخص)تغییر پایا (درمان
66
/
21
88
/
18
11
/
21
42
/
21
درصد بهبودی پس از درمان
درصد بهبودی کلی پس از درمان
5/
55
12
/
79
35
/
80
66
/
86
40
/
75
پیگیری یک ماهه
پیگیری دو ماهه
پیگیری سه ماهه
)شاخص تغییر پایا (درمان
درصد بهبودی پس از درمان
درصد بهبودی کلی پس از درمان
23
23
23
24
/
17
66
/
91
33
33
33
11
/
21
07
/
98
36
36
35
38
/
19
44
/
94
34
33
33
09
/
19
79
/
98
74
/
95 308 308 نمودار2
.روند تغییر نمره های خرده مقیاس امنیت هیجانی
زوج
ها در مراحل خط پایه، مداخله
0
10
20
30
زوج اول
0
10
20
30
40
زوج دوم
0
20
40
خط پایه1
خط پایه2
خط پایه3
خط پایه4
جلسه3
جلسه5
جلسه8
پیگیری1
پیگیری2
پیگیری3
زوج سوم
0
20
40
خط پایه1
خط پایه2
خط پایه3
خط پایه4
خط پایه5
جلسه3
جلسه5
جلسه8
پیگیری1
پیگیری2
پیگیری3
زوج چهارم 309 بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین جدول3) نشان می دهد که مقدار شاخص تغییر پایا در زوج اول، دوم و سوم و چهارم در پایان
درمان به ترتیب66
/
21
،
88
/
18
،
11
/
21
و42
/
21
به دست آمد که با توجه به سطح معناداری
(
05
/0
p>
) تمامی نمرات به باالتر از1.96
z=
رسیده است و بنابراین می توان گفت تغییرات هر
چهار زوج دوم نظر آماری قابل
قبول و نتیجه ی تأثیرات درمانی است. همین طور مقدار شاخص
تغییر پایا در زوج اول، دوم و سوم و چهارم در جلسه ی پیگیری به ترتیب24
/
17
،
11
/
21
،
38
/
19
و09
/
19
به دست آمد که با توجه به( سطح معناداری05
/0
p>) نمر
ۀ
هر چهار زوج به باالتر از
1.96
z=
.رسیده است این میزان بهبودی در هر چهار زوج بیانگر موفقیت و تأثیرات درمانی
می
باشد. بنابراین می توان نتیجه گرفت درمان صمیمیت از درون به برون در افزایش امنیت
هیجانی زوج.ها مؤثر بوده است
جدول4
)نمره های چهار زوج در خرده مقیاس
امنیت خود بودن
را بر حسب مراحل مختلف
درمان را نشان
می.دهد جدول 4. روند تغییر مراحل درمان چهار زوج در خرده مقیاس
امنیت خود بودن
گروه
های درمانی درمان صمیمیت از درون به برون
آزمودنی
ها
مراحل درمان
زوج اول زوج دوم زوج سوم زوج چهارم
میانگین مراحل خط پایه
4
33
/7
75
/8
6/5
میانگین مراحل درمان
33
/8
66
/
14
66
/
15
33
/
14
)شاخص تغییر پایا (درمان
33
/8
87
/7
87
/7
86
/
11
درصد بهبودی پس از درمان
درصد بهبودی کلی پس از
درمان
25
/
108
42
/
109
75
/
95
89
/
155
32
/
117
پیگیری
پیگیری یک ماهه
پیگیری دو ماهه
پیگیری سه ماهه
)شاخص تغییر پایا (درمان
درصد بهبودی پس از درمان
12
11
11
55
/6
175
18
17
17
44
/9
85
/
142
19
19
19
82
/8
5/
137
19
19
19
80
/9
280
درصد بهبودی کلی پس از
درمان
83
/
183 310 فصلنام
ة علمی-
پژوهشی
ن روا
شناسی
،کاربردی ة دور
15
، شمار
ة 2 (
58
)، تابستان
1400
،
317 نمودار3. زوج اول بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین جدول 4) نشان می دهد که مقدار شاخص تغییر پایا در زوج اول، دوم و سوم و چهارم در پایان
درمان به ترتیب33
/8
،
87
/7
،
87
/7 و86
/
11
به دست آمد که با توجه به( سطح معناداری05
/0
p>
) تمامی نمرات به باالتر از1.96
z=
رسیده است و بنابراین می توان گفت تغییرات هر چهار
زوج دوم نظر آماری قابل قبو
ل و نتیج
ۀ
تأثیرات درمانی است. همین طور مقدار شاخص تغییر
پایا در زوج اول، دوم و سوم و چهارم در جلس
ۀ
پیگیری به ترتیب55
/6
،
44
/9
،
82
/8 و80
/9
به
دست آمد که با توجه به( سطح معناداری05
/0
p>) نمره ی هر چهار زوج به باالتر از1.96
z=
.رسیده است این میزان بهبودی در
هر چهار زوج بیانگر موفقیت و تأثیرات درمانی می .باشد
بنابراین می توان نتیجه گرفت درمان صمیمیت از درون به برون در افزایش امنیت خود بودن
زوج.ها مؤثر بوده است هدف از پژوهش حاضر تعیین اثربخشی درمان
صمیمیت از درون به برون (
IFIO
) در بهبود
ارتباط ایمن، امنیت هیجانی و امنیت خود بودن زوج .های شهرستان دزفول بود در این پژوهش
میزان
ارتباط ایمن
.هر چهار زوج در پایان درمان افزایش یافت نتایج پژوهش حاضر نشان داد که
هیچ پژوهش مشابهی تاکنون در داخل و حتی خارج از کشور و در کار با زوجین انجام نشده است
ام
ا یافته های سایر متغیرها با نتایج تحقیقات ،هاگ، جانسن، کریستینسن، فریدریکسن، دالتون
( دیرگرو و همکاران2018)؛
( کناپ و همکاران2015
( )؛ اوکا و همکاران2014
( )؛ باولک2013
)؛
( هامیل2010
( )؛ ترامبلی و سالیوان2010
( )؛ سیدال و وامپلر2013
)؛ میکولینسر و شاوور
(
2007
)
.همخوانی دارد انجام ده ها سال پژوهش و بررسی نشان داده است که وجود یک شخص
و ایجاد حس دلبستگی در بزرگسالی می تواند باعث باال رفتن حس امنیت، راحتی و تأثیر مثبت
،شود (میکولینسر و شاور2007). پژوهش های به عمل آمده بر روی ارتباط ایمن زوجین نشان
می دهند که
رفتارهای نشان دهنده ی دسترسی، پاسخگویی و پذیرندگی با دلبستگی ایمن مرتبط
،است (ساندبرگ، بازبی، جانسون و یوشیدا2012
). درمان صمیمیت از درون به برون با ایجاد
صمیمیت از دست رفته و ایجاد لحظه پیوند و پذیرندگی باعث می شود که زوجین برای رفتن به
برون و شناسایی زخم و جراحات احساس ایمنی کنند. و در نتیجه درمان می تواند بر روی زوجین
تأثیر بگذارد و ارتباط ایمن را ایجاد کند. بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین روند تغییر نمره های در خرده مقیاس
امنیت خود بودن زوج
ها در مراحل خط پایه، مداخله و پ
0
5
10
15
زوج اول
0
5
10
15
20
زوج دوم
0
10
20
خط پایه1
خط پایه2
خط پایه3
خط پایه4
جلسه3
جلسه5
جلسه8
پیگیری1
پیگیری2
پیگیری3
زوج سوم
0
10
20
خط پایه1
خط پایه2
خط پایه3
خط پایه4
خط پایه5
جلسه3
جلسه5
جلسه8
پیگیری1
پیگیری2
پیگیری3
زوج چهارم 311 بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین بنابراین با توجه به یافته
های به دست آمده می توان به
سؤال اول پژوهش این طور پاسخ داد که درمان صمیمیت از درون به برون (
IFIO
) در بهبود
ارتباط ایمن زوج ین مؤثر است. همچنین در پژوهش حاضر میزان امنیت هیجانی هر چهار زوج
در پایان جلس
ۀ پیگیری افزایش یافت. یافته ،های پژوهش حاضر با نتایج تحقیقات باقری، کیمیایی 312 صلنام
ة علمی-
پژوهشی
ن روا
شناسی
،کاربردی ة دور
15
، شمار
ة 2 (
58
)، تابستان
1400
،
317
-
297 ( کارشکی1399
( )؛ فتوت، قهاری و سالمی خامنه1398)؛ جمعه( پور و محمودی پور1397
)؛
سودانی، مومنی جاوید، مهر( ابی زاده هنرمند و خجسته مهر1395
،)؛ هاگ، جانسن، کریستینسن
( فریدریکسن، دالتون، دیرگرو و همکاران2018
( )؛ کریزیکو و تامپسون2015
.) همخوانی دارد
کریسیکو و
( تامسون2015) دریافتند که در روابط بین فردی همسرها عدم پذیرش عاطفی
زوجین توسط همسر (عدم ابراز احساس
ها و نداشتن همدلی) و تقابل عاطفی منفی از عوامل
مؤثر در نارضایتی زناشویی
است. این درمان
پیوند
ی امن را
پرورش
می
د ده که تقویت کنند
ۀ
تعامالت باز، مراقبت از شریک و فهم تفاوت
ها و نیز شباهت
ها
ی همدیگر است. بنابراین، در طول
درمان، زوجین شیو
ۀ جدیدی را در اقرار (درد و دل کردن) به بخش
های آسیب پذیر خود یاد
می
گیرند، چیزی که آن را تساهل ارتباطی می
نامیم. وقتی که یک شریک با بخش های تبعیدی
و آسیب
های درونی خود تماس برقرار می
کند، دیگری هم آن ها را به صورت درونی، احساس
می
کند. هر دو در یک زمان با آن
ها تماس برقرار می کنند، این اقرار به آسیب (درد دل کردن
خود)، به انکارها اجازه می
دهد تا یک تجرب
ۀ
متناقض و اصالحی در درون و بیرون را تجربه کنند
،(اِکر، تیکیس و هالی2012
.)بنابراین با توجه به یافته
های به دست آمده می توان به سؤال دوم
پژوهش این طور پاسخ داد که درمان صمیمیت از درون به برون (
IFIO
) در بهبود امنیت هیجانی
.زوجین مؤثر است
میزان امنیت خود بودن هر چهار زوج نیز در پایان جلس
ۀ
پیگیری افزایش
یافت. بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین و این
یافته( ها با نتایج تحقیقات سودانی و همکاران1395)؛ هربین-
بالنک و همکاران
(
2016
( )؛ گالو1999
؛ به نقل ،از باولک2013
( )؛ میکولینسر و شاوور2007
.) همخوانی دارد
رویکردIFIO
به زوجین در دستیابی به"خود"
هر
دوی آن
ها کمک می کند، این منبع قدرتمند
را به همراه مفاهیم بخش
ها در یک محیط ارتباطی، به عنوان وسیله ای برای رشد و التیام افراد و
زوجین
توسعه می.دهد
از آن
جا که رابط
ۀ بین بخش ها و"خود" باعث وضوح و شفافیت می ،شود
و تجارب
ناگوار و آسیب
زا را التیام می
بخشد، گسترش این رابطه، هدف اصلی درمان
سیستم
( خانواده درونیIFS
)
و همچنینIFIO
.است
از طرفی وقتی که زوجین وارد درمان می ،شوند
در وضعیت قطع ارتباط، خشم و اندوه و
از دست دادن صمیمیت به سر می
برند و بازسازی رابط
ۀ
عاشقانه می
تواند زخم .های کهنه را التیام بخشد امنیت درونی، که درIFIO
دلبستگی با"خود"
نامیده می
شود، در بازگرداندن صلح و امنیت درونی و همچنین امنیت خود بودن
مهم است
(هربین-
،بالنک و همکاران2016
). بنابراین
با توجه به یافته
های به دست آمده می توان به سؤال
سوم پژوهش این طور پاسخ داد که درمان صمیمیت از درون به برون (
IFIO
) در بهبود امنیت 313 بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین با توجه به این که درمان
صمیمیت از درون به برون
در ایران برای درمان مشکالت زوجی به کار
نرفته بود، لذا عدم امکان مقایس
ۀ
نتایج حاصل از تحقیق حاضر با تحقیقات دیگر از جمله
محدودیت .های پژوهش حاضر بود
یکی از محدودیت
های جلس
ۀ
حاضر این بود که به دلیل شیوع
ویروس کرونا به زوجین استرس و فشار مضاعف وارد می کرد که عالوه بر مشکالت بین زوجین
ترس از ابتال و تفاوت زو
جین در میزان رعایت دستورالعمل های بهداشتی هم به تنش بین زوجین
دامن می
زد و گاهی نتایج درمانی را خدشه دار می.کرد با این حال داده
های این مطالع
ۀ
مقدماتی
نشان می دهد کهIFIO
به طور بالقوه در افزایش ارتباط ایمن، امنیت هیجانی و امنیت خود
بودن زوج .ها مؤثر است
پیشنهاد می
شود متخصصین و درمانگرانی که با خانواده
ها و زوج ها در
ایجاد صمیمیت و ارتباط ایمن و سازگار کار می کنند از درمان صمیمیت از درون به برون در
رسیدن به اهداف درمانی خود استفاده کنند. همچنین پیشنهاد می
شود که پژوهش های آینده
کاربرد درمان صمیمیت از درون به برون را با توجه به تفاوت های جنسیتی بین زن و مرد به کار
ببرند تا تأثیر آن به تفکیک روی زوجین مشخص شود. و پژوهش های آینده این درمان را بر روی
.سایر متغیرهای زوجی مثل رضایت زناشویی، کیفیت زناشویی و یا رضایت از همسر به کار برند
امید است که مطالعات آینده ب ر روی نتایج پژوهش حاضر با بررسی کاربردIFIO
در مشکالت
زوج
ها با استفاده از نمونه
هایی بزرگ
تر، با تنوع بیشتر و طرح های پژوهشی با کنترل بیشتر ایجاد
.شود موازین اخالقی
در این پژوهش موازین اخالقی شامل اخذ رضایت
،آگاهانه تضمین حریم خصوصی و
رازداری
رعایت
،شده و به شرکت کنندگان اطمینان داده شد که اطالعات محرمانه می
ماند و این امر
نیز
ًکامال رعایت گشت. همچنین قبل از اجرای پژوهش زوجین چند جلس
ۀ خط پایه و یک جلس
ۀ
توجیهی داشتند که در این جلسات به آن
ها اجازه داده شد در صورت عدم تمایل می توانند تحقیق
.را ترک کنند هواز دانشگاه شهید چمران اهواز.
)(پیوند. سپاسگزاری ا سپاسگزاری
محقق بر خود الزم می
داند تا از زحمات کلیه
ی عزیزانی که در انجام این مهم یاری گر بودند و
همچنین زوج.هایی که در این پژوهش شرکت کردند کمال تشکر و قدردانی را کند 314 فصلنام
ة علمی-
پژوهشی
ن روا
شناسی
،کاربردی ة دور
15
، شمار
ة 2 (
58
)، تابستان
1400
،
317
-
97 مشارکت نویسندگان
این مقاله برگرفته از رسال
ۀ دکتری نویسند
ۀ مسئول پژوهش است. و تمامی نو یسندگان دیگر به
ترتیب استاد راهنما و اساتید مشاور در اجرای این رساله بودند که همگی نقش راهنمایی و نظارت
.بر حسن انجام کار را داشتند ( ایمانی، مهدی1391
). بررسی کارآیی رفتاردرمانی مبتنی بر پذیرش و زوج درمانی رفتاری یکپارچه نگر
.در کاهش آشفتگی زناشویی و عالئم اختالل اضطراب فراگیر در زنان شهر ماهشهر
پایان نامه .ی دکترا
اهواز دانشگاه شهید چمران اهواز. )(پیوند. ( ایمانی، مهدی1391
). بررسی کارآیی رفتاردرمانی مبتنی بر پذیرش و زوج درمانی رفتاری یکپارچه نگر
.در کاهش آشفتگی زناشویی و عالئم اختالل اضطراب فراگیر در زنان شهر ماهشهر
پایان نامه .ی دکترا
اهواز دانشگاه شهید چمران اهواز. )(پیوند. Ainsworth, M. D. S., Blehar, M. C., Waters, E., & Wall, S. (1978). Patterns
of attachment: A psychological study of the strange situation. Hillsdale,
NJ: Erlbaum.(Link). Arnold, E. & Boggs, K. (2015). Professional Communication Skills for
Nurses. Interpersonal
Relationships.7th
Edition. 9780323328579. Saunders. 8th January. Page Count: 576. (Link). Bowlby, J. (1969). Attachment and loss: Vol. 1. Attachment. New York, NY:
Basic Books. (Link). Bowlby, J. (1973). Attachment and loss: Vol. 2. Separation: Anxiety and
anger. New York: Basic Books. (Link). Cordova, J. V., Gee, C. B., & Warren, L. Z. (2005). Emotional skillfulness in
marriage: Intimacy as a mediator of the relationship between emotional
skillfulness and marital satisfaction. Journal of Social and Clinical
Psychology, 24(2), 218–235. (Link). Feeney, J. A. (2002). Attachment, marital interaction, and relationship
satisfaction: A diary study. Personal Relationships, 9(1), 39–55. doi:10.1111/1475-6811.00003. (Link). Feeney, J. A. (2008). Adult romantic attachment: Development in the study of
couple relationships. In J. Cassidy & P. R. Shaver (Eds.), Handbook of
attachment: Theory, research, and clinical applications (pp. 456–481). New York, NY: The Guilford Press. (Link). Hammill, C. (2010). The relationship of childhood maltreatment and adult
style to functional recovery in chronic fatigue syndrome and fibromyalgia. Dissertation Abstracts International Section B, 70, 5820. (Link). 315 Sandberg, J. G., Busby, D. M., Johnson, S. M., & Yoshida, K. (2012). The
brief accessibility, responsiveness, and engagement (BARE) scale: A tool
for measuring attachment behavior in couple relationships. Family
Process, 51, 512–526. (Link). بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین Herbine-Blank, T. Kerpelman, D. & Sweezy, M. (2016). Intimacy From the
Inside Out therapy. Courage and Compassion in Couple Therapy. by
Routledge 711 Third Avenue, New York, NY 10017. (Link). Høeg,B. L., Johansen, C., Christensen, J., Frederiksen, K., Dalton, S. O.,
Dyregrov, A., ... & Bidstrup, P. E. (2018). Early parental loss and intimate
relationships in adulthood: A nationwide study. Developmental
psychology, 54(5), 963. (Link). Johnson, S. M. (2004). The practice of emotionally focused couple therapy:
Creating connection (2nd edn). New York: Brunner-Routledge. (Link). Johnson, S. M. (2008). Hold me tight: Seven conversations for a lifetime of
love. New York: Little, Brown and Company. (Link). Johnson, S., & Greenman, P. (2013). Commentary: Of course it is all about
attachment. Journal of Marital and Family Therapy, 39, 421–423. (Link). Knapp, D. J., Sandberg, J. G., Novak, J., & Larson, J. H. (2015). The
mediating role of attachment behaviors on the relationship between family-
of-origin and couple communication: Implications for couples therapy. Journal of Couple & Relationship Therapy: Innovations in Clinical and
Educational Interventions, 14(1), 17–38. (Link). Koçyiğit Özyiğit, M. (2017). The Meaning of Marriage According to
University Students: A Phenomenological Study. Educational Sciences:
Theory & Practice: Vol. 17 No. 2. (Link). Mcevoy, D. (2005). Attachment styles among violent and non-violent
mentally ill and non-mentally ill jail inmates. Dissertation Abstracts
International Section B, 65, 4839. (Link). Mikulincer, M., & Shaver, P. R. (2007). Attachment in adulthood: Structure,
dynamics, and change. New York: Guilford Press. (Link). Oka, M., Sandberg, J. G., Bradford, A. B., & Brown, A. (2014). Insecure
attachment behavior and partner violence: Incorporating couple
perceptions of insecure attachment and relational aggression. Journal of
Marital and Family Therapy, 40(4), 412–429. (Link). Ponzetti, J. J., & Mutch, B. H. (2006). Marriage as a covenant: Tradition as a
guide to marriage education in the pastoral context. Pastoral Psychology,
54 (3), 215-230. (Link). Reis, S., & Grenyer, B. (2004). Fear of intimacy of women: Relationship
between attachment style and depressive symptoms. Psychopathology, 37,
299–303. (Link). Rholes, W. S., Simpson, J. A., Campbell, L., & Grich, J. (2001). Adult
attachment and the transition to parenthood. Interpersonal Relations and
Group Processes, 81(3), 421–435. doi:10.1037//0022-3514.81.3.421. (Link). 316 Sandberg, J. G., Busby, D. M., Johnson, S. M., & Yoshida, K. (2012). The
brief accessibility, responsiveness, and engagement (BARE) scale: A tool
for measuring attachment behavior in couple relationships. Family
Process, 51, 512–526. (Link). Seedall, R. B., & Wampler, K. Seedall, R. B., & Wampler, K. S. (2013). An attachment primer for couple
therapists: Research and clinical implications. Journal of Marital and
Family Therapy, 39, 427–440. doi:10.1111/jmft.12024. (Link). صلنام
ة علمی-
پژوهشی
ن روا
شناسی
،کاربردی ة دور
15
، شمار
ة 2 (
58
)، تابستان
1400
،
317
-
297 بررسی اثربخشی درمان صمیمیت از درون به برون (
IFIO
)در بهبود ارتباط ایمن زوجین S. (2013). An attachment primer for couple
therapists: Research and clinical implications. Journal of Marital and
Family Therapy, 39, 427–440. doi:10.1111/jmft.12024. (Link). Tremblay, I., & Sullivan, M. (2010). Attachment and pain outcomes in
adolescents: The mediating role of pain catastrophizing and anxiety. The
Journal of Pain, 11, 160–171. (Link). 317 317
|
https://openalex.org/W2252526781
|
https://www.nature.com/articles/ncomms9442.pdf
|
English
| null |
Genetic sharing and heritability of paediatric age of onset autoimmune diseases
|
Nature communications
| 2,015
|
cc-by
| 13,152
|
ARTICLE Received 14 Feb 2015 | Accepted 21 Aug 2015 | Published 9 Oct 2015 1 Center for Applied Genomics, Children’s Hospital of Philadelphia, Philadelphia, Pennsylvania 19104, USA. 2 Medical Scientist Training Program, Perelman School of Medicine, University of Pennsylvania,
Philadelphia, Pennsylvania 19104, USA. 3 Department of Statistics, University of Illinois at Urbana-Champaign, Champaign, Illinois 61820, USA. 4 Department of Biostatistics and Epidemiology, Perelman
School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA. 5 Department of Computer Science, New Jersey Institute of Technology, Newark, New Jersey 07103, USA.
6 Division of Allergy and Immunology, Children’s Hospital of Philadelphia, Philadelphia, Pennsylvania 19104, USA. 7 Department of Rheumatology, Oslo University Hospital, Rikshospitalet, Oslo 0372,
Norway. 8 Center for Inflammatory Bowel Disease, Division of Gastroenterology, Cincinnati Children’s Hospital Medical Center, Cincinnati, Ohio 45229, USA. 9 Divison of Rheumatology, Cincinnati
Children’s Hospital Medical Center, Cincinnati, Ohio 45229, USA. 10 Division of Rheumatology and Division of Clinical Pharmacology, Toxicology, and Therapeutic Innovation, Children’s Mercy-Kansas
City, Kansas City, Missouri 64108, USA. 11 Department of Pediatrics, University of Utah School of Medicine and Primary Children’s Medical Center, Salt Lake City, Utah 84113, USA. 12 RCCS ‘Casa Sollievo
della Sofferenza’, Division of Gastroenterology, San Giovanni Rotondo 71013, Italy. 13 Division of Pediatric Allergy and Immunology, University of Miami Miller School of Medicine, Miami, Florida 33136,
USA. 14 Institute of Immunology, Department of Medicine, Icahn School of Medicine at Mount Sinai, Mount Sinai Hospital, New York, New York 10029, USA. 15 Department of Translational Medical
Science, Section of Pediatrics, University of Naples "Federico II", Naples 80138, Italy. 16 IBD Centre, Mount Sinai Hospital, University of Toronto, 441-600 University Avenue, Toronto, Ontario, Canada
M5G 1X5. 17 Department of Immunology, Oslo University Hospital, Rikshospitalet, 0027 Oslo 0372, Norway. 18 Texas Scottish Rite Hospital for Children, Dallas, Texas 750219, USA. 19 Yorkhill Hospital
for Sick Children, Glasgow G38SJ, Scotland. 20 Paediatric Gastroenterology and Nutrition, Royal Hospital for Sick Children, Edinburgh and Child Life and Health, University of Edinburgh, Edinburgh EH9
1UW, UK. 21 Departments of Pediatrics and Common Disease Genetics, Cedars Sinai Medical Center, Los Angeles, California 90048, USA. 22 Department of Pathology, Children’s Hospital of
Philadelphia, Philadelphia, Pennsylvania 19104, USA. 23 Department of Pediatrics, The Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA. 24 Unit of
Gastroenterology, Department of Medical and Surgical Specialties, Careggi University Hospital, Viale Pieraccini 18, Florence 50139, Italy. 25 Paediatric Rheumatology Unit, Royal Children’s Hospital,
Parkville, Victoria 3052, Australia. 26 Arthritis and Rheumatology Research, Murdoch Childrens Research Institute, Parkville, Victoria 3052, Australia. 27 Sarah M. and Charles E. Seay Center for
Musculoskeletal Research, Texas Scottish Rite Hospital for Children, Dallas, Texas 750219, USA. 28 Department of Clinical Immunology, Nuffield Department of Medicine, University of Oxford, OX1 1NF,
UK. 29 Section of Immunology, Allergy, and Rheumatology, Department of Pediatric Medicine, Texas Children’s Hospital, Houston, Texas 77030, USA. 30 Division of Rheumatology, Children’s Hospital of
Philadelphia, Philadelphia, Pennsylvania 19104, USA. 31 Department of Pediatrics, Emory University School of Medicine and Children’s Health Care of Atlanta, Atlanta, Georgia 30329, USA. 32 Hospital for
Sick Children, University of Toronto, 555 University Avenue, Toronto, Ontario, Canada M5G 1X8. 33 Gastrointestinal Unit, Division of Medical Sciences, School of Molecular and Clinical Medicine,
University of Edinburgh, Western General Hospital, Edinburgh EH4 2XU, UK. 34 Department of Pediatrics, Nemours Children’s Hospital, Orlando, Florida 32827, USA. 35 Departments of Pediatrics and
Human Genetics, McGill University, Montreal, Quebec, Canada H3H 1P3. 36 Division of Gastroenterology, Children’s Hospital of Philadelphia, Philadelphia, Pennsylvania 19104, USA. 37 Department of
Pathology and Lab Medicine, Perelman School of Medicine University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA. 38 Genes, Environment and Complex Disease, Murdoch Childrens Research
Institute, Parkville, Victoria 3052, Australia. 39 Department of Paediatrics, University of Melbourne, Parkville, Victoria 3052, Australia. 40 Division of Pulmonary Medicine, Children’s Hospital of
Philadelphia, Philadelphia, Pennsylvania 19104, USA. Correspondence and requests for materials should be addressed to H.H. (email: hakonarson@email.chop.edu) & 2015 Macmillan Publishers Limited. All rights reserved. NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 Heritability, in the broad-sense (H2), is defined as the entirety of
an
individual’s
phenotypic
variation
explained
by
genetic
variance, but in practicality, it can be difficult to quantify and
partition precisely15. A major contribution to H2 is the narrow-
sense or additive heritability (h2), which can be more accurately
quantified.15. Recently, a new method was established to estimate
the total phenotype variance attributable to additive genetic
variations using genome-wide SNP genotyping data16–19. The
method has been since applied to dozens of GWAS-examined
traits and extended to examine jointly the co-heritability of
related diseases20. We used a previously described method for estimating disease
variance explained by additive genetic factors using GWAS data
(referred
to
as
SNP-based
heritability
or
SNP-h2)17. We
transformed the SNP-h2 estimates from the observed to the
liability-scale using respective observed disease prevalence. To
assess if our SNP-h2 estimates are consistent with previously
published
findings
and
other
population-based
heritability
estimates (POP-h2), we performed a systematic literature search
followed by manual curation of prevalence and heritability
estimates for each of the nine pAIDs (Fig. 1a and Supplementary
Tables 1 and 2). Among the pAIDs examined where the SNP-h2 estimates were
at least nominally significant (Po0.05), T1D and juvenile
idiopathic
arthritis
(JIA)
were
the
most
highly
heritable
(Fig. 1b). Considerably lower estimates were observed for
ulcerative colitis (UC) and Crohn’s disease (CD; Supplementary
Fig. 1A), suggesting that environmental factors may play a much
larger role in IBD aetiology (Fig. 1d). We also observed relatively
low SNP-h2 estimates for systemic lupus erythematosus (SLE;
0.205±s.e. 0.076). We systematically quantified the narrow-sense heritability, h2,
as well as the pairwise joint heritability of pAIDs attributable to
common genomic variation using a single-centre accrued cohort
of over 5,000 unrelated cases composed of nine independent
pAIDs and 36,000 shared, population-based healthy controls. We
first report the genome-wide SNP genotype-derived heritability
estimates (referred to as SNP-h2) and then the genetic correlation
(SNP-rG) across pairs of the nine investigated pAIDs. We
contextualize these findings alongside a comprehensive review of
available literature and epidemiological data sets, illustrate a
method for quantifying genetic risk factor sharing across pAIDs,
and provide considerations for how such genetic data can aid
disease prediction. Contribution of the MHC region and ChrX to SNP-h2. Given
the known association of variants across the MHC with AI
diseases, we quantified their contribution to the SNP-h2 for each
of the nine pAIDs. NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 Table 1 | Summary of cohorts included. Disease
Full disease name
Cases
Controls
GIF*
Prevalencew
CD
Crohn’s disease
1,848
27,457
1.086
3.00E 03
CEL
Celiac disease
137
27,435
1.006
1.00E 02
CVID
Common variable
immunodeficiency
disorder
304
27,492
1.010
1.00E 04
EPI
Epilepsy
754
26,122
1.027
1.00E 04
JIA
Juvenile idiopathic
arthritis
1,112
27,131
1.000
2.00E 03
PS
Psoriasis
85
27,474
1.012
1.00E 03
SLE
Systemic lupus
erythematosus
252
27,525
1.019
1.00E 04
SPA
Spondyloarthropathy
98
27,483
1.020
1.00E 04
T1D
Type 1 diabetes
664
27,395
1.062
5.00E 03
UC
Ulcerative colitis
854
27,482
1.041
1.00E 03
GIF, genomic inflation factor; SNP, single-nucleotide polymorphism. *GIF is provided for each cohort and included all SNPs (including ChrX and the extended MHC). wPrevalence estimates used here are those made based on observations at our center. A A
utoimmune (AI) diseases affect approximately 1 in 12
individuals living in the Western Hemisphere, represent-
ing a significant cause of morbidity, chronic disability and
health-care burden. High rates of sibling recurrence and twin–
twin concordance, both within and across multiple independent
AI diseases, coupled with recent results from genome-wide
association studies (GWAS), suggest that a set of shared genetic
risk factors underlie paediatric AI disease (pAID) aetiology1–3. Moreover, a number of AI diseases show clear familial clustering,
such as inflammatory bowel disease (IBD)4, whereas others (for
example, type 1 diabetes (T1D), AI thyroiditis (THY) and celiac
disease
(CEL)
may
manifest
as
comorbid
diseases
in
polyglandular AI syndromes2. Although the concept of genetic
sharing among AIs is intriguing, it remains unclear if this is due
to ‘pleiotropic’ risk factors that predispose to multiple AI diseases
via shared mechanisms or if multiple, independent risk factors are
responsible. Table 1 | Summary of cohorts included. GIF, genomic inflation factor; SNP, single-nucleotide polymorphism. *GIF is provided for each cohort and included all SNPs (including ChrX and the extended MHC). wPrevalence estimates used here are those made based on observations at our center. p
GWAS
have
identified
single-nucleotide
polymorphisms
(SNPs) across hundreds of loci as being associated with an
increased risk of developing AI5–12. These findings, coupled with
those from epidemiological studies, strongly support the existence
of (i) an overlapping ‘AI disease genetic landscape’13,14 and
(ii), consequently, a shared heritability across these diseases. NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 We first performed HLA imputation21, to
identify the most strongly associated SNP, amino acid or HLA
allele with each pAID (Supplementary Table 5) and we estimated
POP-h2 attributable to the extended MHC based on previous
analyses (Supplementary Tables 6 and 7). The MHC-specific
SNP-h2 estimates correlated well with the strength of lead MHC
P-value. For example, variations across the extended MHC region
accounted for 32.7% of the total autosomal SNP-h2 in T1D
and 24.7% of that in CEL, with no significant contribution to the
SNP-h2 estimates in psoriasis (PS), SLE, CD or the non-pAID,
EPI. Despite the pervasive association between SNPs within the
MHC and both JIA and UC, contributions of the extended MHC
to their total SNP-h2 (10.7% and 5.8%, respectively) were limited
(Fig. 1c and Table 2). Despite the known association with
HLA-DRB1*0103 and HLA-B*52 in UC13, we observed that
removing the extended MHC did not significantly reduce the
observed SNP-h2 for either UC or, the related IBD phenotype, CD
(Supplementary Table 8). As expected, the contribution of ChrX
to the overall SNP-h2 was small across all pAIDs (Supplementary
Table 2). These estimates are consistent with expectations as NATURE COMMUNICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | www.nature.com/naturecommunications
& 2015 Macmillan Publishers Limited. All rights reserved. Genetic sharing and heritability of paediatric
age of onset autoimmune diseases Yun R. Li1,2, Sihai D. Zhao3, Jin Li1, Jonathan P. Bradfield1, Maede Mohebnasab1, Laura Steel1, Julie Kobie4, Debra J. Abrams1,
Frank D. Mentch1, Joseph T. Glessner1, Yiran Guo1, Zhi Wei1,5, John J. Connolly1, Christopher J. Cardinale1, Marina Bakay1,
Dong Li1, S. Melkorka Maggadottir1,6, Kelly A. Thomas1, Haijun Qui1, Rosetta M. Chiavacci1, Cecilia E. Kim1, Fengxiang Wang1,
James Snyder1, Berit Flatø7, Øystein Førre7, Lee A. Denson8, Susan D. Thompson9, Mara L. Becker10, Stephen L. Guthery11,
Anna Latiano12, Elena Perez13, Elena Resnick14, Caterina Strisciuglio15, Annamaria Staiano15, Erasmo Miele15, Mark S. Silverberg16,
Benedicte A. Lie17, Marilynn Punaro18, Richard K. Russell19, David C. Wilson20, Marla C. Dubinsky21, Dimitri S. Monos22,23,
Vito Annese24, Jane E. Munro25,26, Carol Wise27, Helen Chapel28, Charlotte Cunningham-Rundles14, Jordan S. Orange29,
Edward M. Behrens23,30, Kathleen E. Sullivan6,23, Subra Kugathasan31, Anne M. Griffiths32, Jack Satsangi33, Struan F.A. Grant1,23,
Patrick M.A. Sleiman1,23, Terri H. Finkel34, Constantin Polychronakos35, Robert N. Baldassano23,36, Eline T. Luning Prak37,
Justine A. Ellis38,39, Hongzhe Li4, Brendan J. Keating1,23 & Hakon Hakonarson1,23,40 Autoimmune diseases (AIDs) are polygenic diseases affecting 7–10% of the population in the Western
Hemisphere with few effective therapies. Here, we quantify the heritability of paediatric AIDs (pAIDs),
including JIA, SLE, CEL, T1D, UC, CD, PS, SPA and CVID, attributable to common genomic variations (SNP-
h2). SNP-h2 estimates are most significant for T1D (0.863±s.e. 0.07) and JIA (0.727±s.e. 0.037), more
modest for UC (0.386±s.e. 0.04) and CD (0.454±0.025), largely consistent with population estimates
and are generally greater than that previously reported by adult GWAS. On pairwise analysis, we observed
that the diseases UC-CD (0.69±s.e. 0.07) and JIA-CVID (0.343±s.e. 0.13) are the most strongly cor-
related. Variations across the MHC strongly contribute to SNP-h2 in T1D and JIA, but does not significantly
contribute to the pairwise rG. Together, our results partition contributions of shared versus disease-specific
genomic variations to pAID heritability, identifying pAIDs with unexpected risk sharing, while recapitulating
known associations between autoimmune diseases previously reported in adult cohorts. 1 NATURE COMMUNICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | www.nature.com/naturecommunications & 2015 Macmillan Publishers Limited. All rights reserved. ARTICLE ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 0.00
0.10
0.20
0.30
0.40
0.50
0.60
0.70
0.80
0.90
1.00
CD
CEL CVID JIA
PS
SLE SPA T1D
UC
0.0%
0.1%
0.2%
0.3%
0.4%
0.5%
0.6%
0.7%
0.8%
0.9%
1.0%
POP-h2
Prevalence
Mean prevalence
POP-h2
JIA
UC
T1D
CD
0.0
0.3
0.5
0.8
1.0
CD
CEL CVID EPI
JIA
SLE SPA T1D
UC
Esimated H2T
MHC SNP-h2
MHC SNP-h2 (LIT)
MHC SNP-h2 (GWAS)
SNP-h2
0.0
0.3
0.5
0.8
1.0
CD
CEL CVID
EPI
JIA
SLE
SPA
T1D
UC
Estimated H2
SNP-h2
POP-h2
SNP-h2 (lit)
a
b
c
d
exMHC,
36.3%
ChrX, 1.2%
MHC, 2.1%
exMHC,
60.1%
ENV,
21.1%
MHC,
7.6%
ChrX, 3.1%
ENV,
53.3%
ChrX, 2.8%
MHC,
27.7%
ChrX, 1.4%
MHC,
0.5%
exMHC,
68.2%
ENV,
60.4%
exMHC,
44.7%
ENV, 9.4%
exMHC
ChrX
MHC
ENV
Figure 1 | Autoimmune disease prevalence and heritability estimates. (a) Mean population-based AI disease prevalence (orange) and heritability (blue
estimates (mean±s.d.). Data are curated from epidemiological surveys among Caucasian populations in Europe or North America based on studies
indexed in PubMed between 1975 and 2015. Where multiple sources of data are available for a given trait, we reported a simple non-weighted arithmeti
mean and provided as error bars the standard deviation. Most heritability estimates were based on twin concordance rates. Raw data used and reference
can be found in Supplementary Tables 1 and 2. (b) Univariate SNP-heritability (SNP-h2, orange) compared with estimates reported by prior studies. (SNP-h
(lit), blue) based on variations across the autosomes compared with population-based estimates (POP-h2, red) as reported in the literature (lit). Raw dat
used from prior GWAS SNP-h2 estimates are provided in Supplementary Table 3. Error bars denote standard error. (c) Univariate SNP-heritability
(autosomal) estimates with (Light green, wide) and without the extended MHC (orange, narrow). Results are compared with corresponding heritability
estimates reported using population-based (red, narrow) versus other published SNP-heritability estimates (blue, narrow), when available for a given
disease. Literature data used and references can be found in Supplementary Table 2 and Supplementary Tables 6 and 7. Error bars denote standard erro
(d) Partitioning phenotypic variance to genetic and non-genetic (ENV, green) components in the four largest pAID cohorts. Genetic components includ
contributions from the entire autosomal regions excluding the MHC (exMHC, orange), the extended MHC (MHC, blue) alone as well as from the
X-chromosome (ChrX, red). NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 (a) Mean population-based AI disease prevalence (orange) and heritability (blue) ion of autosomal, autosomal with extended MHC removed (exMHC) and ChrX variations to pAID Table 2 | Contribution of autosomal, autosomal with extended MHC removed (exMHC) and ChrX variations to pAID
heritability (h2). Disease
h2(auto)
s.e. P
h2(exMHC)
s.e. P
%MHC*
ChrX
s.e. P
CD
0.454
0.025
o1.00E–04
0.447
0.025
o1.00E–04
1.54
0.014
0.004
2.35E–04
CEL
0.447
0.362
1.06E–01
0.337
0.361
1.74E–01
24.72
0.048
0.058
1.89E–01
CVIDw
0.181
0.063
1.72E–03
0.167
0.063
3.66E–03
8.12
NA
NA
NA
EPI
0.168
0.027
1.05E–10
0.163
0.027
3.90E–10
2.91
0.010
0.005
1.21E–02
JIA
0.727
0.037
o1.00E–04
0.650
0.037
o1.00E–04
10.66
0.027
0.007
4.91E–06
PS
0.949
0.381
5.90E–03
0.949
0.380
5.87E–03
0.02
0.003
0.061
4.82E–01
SLE
0.206
0.076
3.16E–03
0.202
0.076
3.74E–03
1.89
0.013
0.013
1.60E–01
SPA
0.370
0.192
2.45E–02
0.310
0.191
4.91E–02
16.17
0.029
0.028
1.74E–01
T1D
0.863
0.070
o1.00E–04
0.581
0.069
o1.00E–04
32.66
0.028
0.012
5.27E–03
UC
0.386
0.041
o1.00E–04
0.363
0.041
o1.00E–04
5.84
0.012
0.007
3.38E–02
CD, crohn’s disease; CEL, celiac disease; CVID, common variable immunodeficiency disorder; EPI, epilepsy; JIA, juvenile idiopathic arthritis; NA, not applicable; pAID, paediatric autoimmune disease; PS,
psoriasis; SLE, systemic lupus erythematosus; SPA, spondyloarthropathy; T1D, type 1 diabetes; UC, ulcerative colitis. P-values (P) are based on results from the restricted maximum likelihood estimate (likelihood ratio test). Error bars represent standard error. *Percentage contribution of the extended MHC to total autosomal SNP-h2
wREML estimates could not be made due to limited common SNP variability among this cohort on the X-chromosome Table 2 | Contribution of autosomal, autosomal with extended MHC removed (exMHC) and ChrX variations to pAID
heritability (h2). 2 | Contribution of autosomal, autosomal with extended MHC removed (exMHC) and ChrX variation
bility (h2). CD, crohn’s disease; CEL, celiac disease; CVID, common variable immunodeficiency disorder; EPI, epilepsy; JIA, juvenile idiopathic arthritis; NA, not applicable; pAID, paediatric autoimmune disease; PS,
psoriasis; SLE, systemic lupus erythematosus; SPA, spondyloarthropathy; T1D, type 1 diabetes; UC, ulcerative colitis. P-values (P) are based on results from the restricted maximum likelihood estimate (likelihood ratio test). Error bars represent standard error. *Percentage contribution of the extended MHC to total autosomal SNP-h2
wREML estimates could not be made due to limited common SNP variability among this cohort on the X-chromosome ChrX makes up only about 5% of the total genome22, has
comparatively fewer coding bases and is less polymorphic23. & 2015 Macmillan Publishers Limited. All rights reserved. Results
Q
f Quantifying the heritability of paediatric AI diseases. To
quantify the SNP-h2 of the nine pAIDs, we utilized genome-wide
SNP genotypes ascertained from DNA samples of patients of
each pAID cohort along with samples from population-based
control subjects with no known diagnosis of autoimmunity
or immunodeficiency. Following extensive quality control (QC),
removing
SNPs
of
lower
minor
allele
frequency
(MAF),
missingness and differential missingness in cases and controls,
and deviation from Hardy–Weinberg equilibrium (see Methods),
we retained 461,301 SNPs. We excluded samples for low geno-
typing rates, cryptic relatedness and genetic outliers, leaving a
cohort consisting of 4,956 cases distributed across nine pAIDs
and 27,451 unrelated shared population-based controls (Table 1). We also included, for comparison, a non-immune-mediated
dichotomous trait, paediatric-onset epilepsy (EPI); this cohort of
B800 case subjects was recruited and genotyped at our centre
using the same platforms over the same time period. NATURE COMMUNICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | www.nature.com/naturecommunications
& 2015 Macmillan Publishers Limited. All rights reserved. 2 NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 0.00
0.10
0.20
0.30
0.40
0.50
0.60
0.70
0.80
0.90
1.00
CD
CEL CVID JIA
PS
SLE SPA T1D
UC
0.0%
0.1%
0.2%
0.3%
0.4%
0.5%
0.6%
0.7%
0.8%
0.9%
1.0%
POP-h2
Prevalence
Mean prevalence
POP-h2
a 0.0
0.3
0.5
0.8
1.0
CD
CEL CVID EPI
JIA
SLE SPA T1D
UC
Esimated H2T
MHC SNP-h2
MHC SNP-h2 (LIT)
MHC SNP-h2 (GWAS)
SNP-h2
c a Prevalence 0.0
0.3
0.5
0.8
1.0
CD
CEL CVID
EPI
JIA
SLE
SPA
T1D
UC
Estimated H2
SNP-h2
POP-h2
SNP-h2 (lit)
b b JIA
CD
d
ENV,
21.1%
MHC,
7.6%
ChrX, 3.1%
ENV,
53.3%
ChrX, 1.4%
MHC,
0.5%
exMHC,
68.2%
exMHC,
44.7% J
d d UC
T1D
exMHC,
36.3%
ChrX, 1.2%
MHC, 2.1%
exMHC,
60.1%
ChrX, 2.8%
MHC,
27.7%
ENV,
60.4%
ENV, 9.4%
exMHC
ChrX
MHC
ENV Figure 1 | Autoimmune disease prevalence and heritability estimates. (a) Mean population-based AI disease prevalence (orange) and heritability (blue)
estimates (mean±s.d.). Data are curated from epidemiological surveys among Caucasian populations in Europe or North America based on studies
indexed in PubMed between 1975 and 2015. Where multiple sources of data are available for a given trait, we reported a simple non-weighted arithmetic
mean and provided as error bars the standard deviation. Most heritability estimates were based on twin concordance rates. Raw data used and references
can be found in Supplementary Tables 1 and 2. (b) Univariate SNP-heritability (SNP-h2, orange) compared with estimates reported by prior studies. (SNP-h2
(lit), blue) based on variations across the autosomes compared with population-based estimates (POP-h2, red) as reported in the literature (lit). Raw data
used from prior GWAS SNP-h2 estimates are provided in Supplementary Table 3. Error bars denote standard error. (c) Univariate SNP-heritability
(autosomal) estimates with (Light green, wide) and without the extended MHC (orange, narrow). Results are compared with corresponding heritability
estimates reported using population-based (red, narrow) versus other published SNP-heritability estimates (blue, narrow), when available for a given
disease. Literature data used and references can be found in Supplementary Table 2 and Supplementary Tables 6 and 7. Error bars denote standard error. (d) Partitioning phenotypic variance to genetic and non-genetic (ENV, green) components in the four largest pAID cohorts. Genetic components include
contributions from the entire autosomal regions excluding the MHC (exMHC, orange), the extended MHC (MHC, blue) alone as well as from the
X-chromosome (ChrX, red). evalence and heritability estimates. NICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | www.nature.com/naturecommunications NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 (a) Observed prevalence of pAID Figure 2 | Prevalence of AI disease co-morbidities and estimates of genetic correlation (co-heritability) across pAIDs. (a) Observed prevalence of pAID
comorbidity observed in Caucasian populations in Europe and North America as curated from large-scale cohort studies. For each pairwise combination
(for example, Disease 1–Disease 2), the rate (y axis) indicates the percentage of patients with Disease 2 who have also been diagnosed with Disease 1. Literature data used and references can be found in Supplementary Table 9. (b) Bivariate estimates of genetic correlation (pairwise co-heritability) across
pAIDs. The heritability (SNP-h2) for the first and second disease are shown for each pAID pair (blue and green bars, respectively) along with the genetic
correlation (rG) for the pair estimated based on total autosomal common genetic variants (orange) and based on autosomal variants excluding the MHC
(red). Displayed are those pairs for which the rG estimates reached nominal significance (Po0.05). P-values are based on restricted maximum likelihood
ratio test. Error bars represent standard error. (c) Genetic sharing using the genome-wide pairwise sharing statistic (GPS). Correlation plot of the P-values
obtained from the genome-wide pairwise shared analysis. Significant P-values support evidence of genetic sharing based on the correlation of significant
association findings reported by GWAS for each pair of diseases. of heritability, sample size and the number of causal variants. We assessed the mean and maximum area under the receiver
operating characteristic curve (AUC) achieved, showing that
our SVM predictor was most effective for JIA and T1D
(AUCmax40.9; AUCmean40.85), although satisfactory results
was also seen in CEL (AUCmax40.8 and AUCmean40.7). These
findings are consistent with that recently reported by Speed et al. using an independent adult CEL cohort26. The predictability of all
nine pAIDs was fairly robust to range of P-value thresholds used
for selecting SNP predictors in building the SVM model (Fig. 3
and Supplementary Table 11). and between each of the nine pAIDs and EPI, which provided a
comparative baseline for non-significant genetic correlation20. We used both a strict (PBS) and a more relaxed Bonferroni
correction
(PBL)
to
adjust
for
either
45
(all
pairwise
combinations) or 9 comparisons (combinations per pAID); (see
Methods). We observed the highest rG between UC and CD
(rG ¼ þ 0.66; PBSo0.001), consistent with the reported sharing
of association loci by several published GWAS, immunochip and
fine-mapping studies11,27–29 (Supplementary Table 10). NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 Pairwise correlation or H2 (per disease)
1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0
–0.2
–0.4
–0.6
CVID-JIA EPI-JIA
EPI-UC PSOR-T1D PS-UC
SLE-CD
SPA-CD
T1D-CD
UC-CD
D1 SNP-h2
Autosomal rG
exMHC rG
D2 SNP-h2
b 0
0.88
1.77
2.65
3.54
4.42
CD
JIA
T1D
UC
CVID
SLE
CEL
PS
SPA
THY
SPA
PSOR
CEL
SLE
CVID
UC
T1D
JIA
c b Figure 2 | Prevalence of AI disease co-morbidities and estimates of genetic correlation (co-heritability) across pAIDs. (a) Observed prevalence of pAID
comorbidity observed in Caucasian populations in Europe and North America as curated from large-scale cohort studies. For each pairwise combination
(for example, Disease 1–Disease 2), the rate (y axis) indicates the percentage of patients with Disease 2 who have also been diagnosed with Disease 1. Literature data used and references can be found in Supplementary Table 9. (b) Bivariate estimates of genetic correlation (pairwise co-heritability) across
pAIDs. The heritability (SNP-h2) for the first and second disease are shown for each pAID pair (blue and green bars, respectively) along with the genetic
correlation (rG) for the pair estimated based on total autosomal common genetic variants (orange) and based on autosomal variants excluding the MHC
(
d) Di
l
d
th
i
f
hi h th
G
ti
t
h d
i
l i
ifi
(P
0 05) P
l
b
d
t i t d
i
lik lih
d Figure 2 | Prevalence of AI disease co-morbidities and estimates of genetic correlation (co-heritability) across pAIDs. (a) Observed prevalence of pAID
comorbidity observed in Caucasian populations in Europe and North America as curated from large-scale cohort studies. For each pairwise combination
(for example, Disease 1–Disease 2), the rate (y axis) indicates the percentage of patients with Disease 2 who have also been diagnosed with Disease 1. Literature data used and references can be found in Supplementary Table 9 (b) Bivariate estimates of genetic correlation (pairwise co heritability) across Figure 2 | Prevalence of AI disease co-morbidities and estimates of genetic correlation (co-heritability) across pAID Figure 2 | Prevalence of AI disease co-morbidities and estimates of genetic correlation (co-heritability) across pAIDs. (a) Observed prevalence of pAID
comorbidity observed in Caucasian populations in Europe and North America as curated from large scale cohort studies For each pairwise combination Figure 2 | Prevalence of AI disease co-morbidities and estimates of genetic correlation (co-heritability) across pAIDs. NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 Significant P-values support evidence of genetic sharing based on the correlation of significan
i i
fidi
d b
GWAS f
h
i
f di
ARTICLE
NATURE COMMUNICATIONS | DOI: 10.1038/ncomms944 0.25
0.20
0.15
0.10
0.05
0.00
Prevalence of subjects w/ D1 among Pts with D2
CD-UC
UC-CD
CVID-JIA
CEL-PS
T1D-CEL
CD-SPA
PS-UC
JIA-CEL
PS-T1D
PS-CD
UC-SPA
SLE-T1D
CEL-T1D
CD-PS
UC-PS
CVID-CD
SLE-UC
UC-T1D
CEL-SLE
CEL-CD
SLE-PS
CD-T1D
T1D-JIA
JIA-T1D
CVID-SLE
T1D-PS
CEL-UC
T1D-UC
CD-CEL
SLE-CD
CEl-SPA
T1D-CD
UC-CEL
PS-CEL
PS-SLE
S:E-CEL
CD-SLE
UC-SLE
T1D-SLE
CEL-JIA
a Disease 1 – Disease 2
CD
UC
CVID
CEL
T1D-C
CD-S
PS
JIA-C
PS-T
PS
UC-S
SLE-T
CEL-T
CD
UC
CVID
SLE
UC-T
CEL-S
CEL
SLE
CD-T
T1D
JIA-T
CVID-S
T1D
CEL
T1D
CD-C
SLE
CEl-S
T1D
UC-C
PS-C
PS-S
S:E-C
CD-S
UC-S
T1D-S
CEL
Pairwise correlation or H2 (per disease)
1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0
–0.2
–0.4
–0.6
CVID-JIA EPI-JIA
EPI-UC PSOR-T1D PS-UC
SLE-CD
SPA-CD
T1D-CD
UC-CD
D1 SNP-h2
Autosomal rG
exMHC rG
D2 SNP-h2
0
0.88
1.77
2.65
3.54
4.42
CD
JIA
T1D
UC
CVID
SLE
CEL
PS
SPA
THY
SPA
PSOR
CEL
SLE
CVID
UC
T1D
JIA
b
c
Figure 2 | Prevalence of AI disease co-morbidities and estimates of genetic correlation (co-heritability) across pAIDs. (a) Observed prevalence of pAID
comorbidity observed in Caucasian populations in Europe and North America as curated from large-scale cohort studies. For each pairwise combination
(for example, Disease 1–Disease 2), the rate (y axis) indicates the percentage of patients with Disease 2 who have also been diagnosed with Disease 1. Literature data used and references can be found in Supplementary Table 9. (b) Bivariate estimates of genetic correlation (pairwise co-heritability) across
pAIDs. The heritability (SNP-h2) for the first and second disease are shown for each pAID pair (blue and green bars, respectively) along with the genetic
correlation (rG) for the pair estimated based on total autosomal common genetic variants (orange) and based on autosomal variants excluding the MHC
(red). Displayed are those pairs for which the rG estimates reached nominal significance (Po0.05). P-values are based on restricted maximum likelihood
ratio test. Error bars represent standard error. (c) Genetic sharing using the genome-wide pairwise sharing statistic (GPS). Correlation plot of the P-values
obtained from the genome-wide pairwise shared analysis. Significant P-values support evidence of genetic sharing based on the correlation of significant
association findings reported by GWAS for each pair of diseases. NATURE COMMUNICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | www.nature.com/naturecommunications
& 2015 Macmillan Publishers Limited. All rights reserved. NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 variations in predicting pAID disease risk, using a SVM model-
based approach. Using a tenfold cross-validation study design, we
built a linear SVM model using the top GWAS signals observed
using nine out of ten of the total samples and tested this
SVM predictor in the remaining 10% of the samples. Based on
previous analyses in both case–control24 and quantitative traits25,
we expect that disease prediction accuracy to behave as function Disease prediction using support vector machines (SVM)s. Given that we observed relatively high rates of heritability across
many of the pAIDs, we evaluated the utility of common genomic NATURE COMMUNICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | www.nature.com/naturecommunications 3 0.25
0.20
0.15
0.10
0.05
0.00
Prevalence of subjects w/ D1 among Pts with D2
Disease 1 – Disease 2
CD-UC
UC-CD
CVID-JIA
CEL-PS
T1D-CEL
CD-SPA
PS-UC
JIA-CEL
PS-T1D
PS-CD
UC-SPA
SLE-T1D
CEL-T1D
CD-PS
UC-PS
CVID-CD
SLE-UC
UC-T1D
CEL-SLE
CEL-CD
SLE-PS
CD-T1D
T1D-JIA
JIA-T1D
CVID-SLE
T1D-PS
CEL-UC
T1D-UC
CD-CEL
SLE-CD
CEl-SPA
T1D-CD
UC-CEL
PS-CEL
PS-SLE
S:E-CEL
CD-SLE
UC-SLE
T1D-SLE
CEL-JIA
Pairwise correlation or H2 (per disease)
1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0
–0.2
–0.4
–0.6
CVID-JIA EPI-JIA
EPI-UC PSOR-T1D PS-UC
SLE-CD
SPA-CD
T1D-CD
UC-CD
D1 SNP-h2
Autosomal rG
exMHC rG
D2 SNP-h2
0
0.88
1.77
2.65
3.54
4.42
CD
JIA
T1D
UC
CVID
SLE
CEL
PS
SPA
THY
SPA
PSOR
CEL
SLE
CVID
UC
T1D
JIA
a
b
c
Figure 2 | Prevalence of AI disease co-morbidities and estimates of genetic correlation (co-heritability) across pAIDs. (a) Observed prevalence of pA
comorbidity observed in Caucasian populations in Europe and North America as curated from large-scale cohort studies. For each pairwise combinatio
(for example, Disease 1–Disease 2), the rate (y axis) indicates the percentage of patients with Disease 2 who have also been diagnosed with Disease
Literature data used and references can be found in Supplementary Table 9. (b) Bivariate estimates of genetic correlation (pairwise co-heritability) acro
pAIDs. The heritability (SNP-h2) for the first and second disease are shown for each pAID pair (blue and green bars, respectively) along with the genet
correlation (rG) for the pair estimated based on total autosomal common genetic variants (orange) and based on autosomal variants excluding the MH
(red). Displayed are those pairs for which the rG estimates reached nominal significance (Po0.05). P-values are based on restricted maximum likelihoo
ratio test. Error bars represent standard error. NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 (c) Genetic sharing using the genome-wide pairwise sharing statistic (GPS). Correlation plot of the P-valu
obtained from the genome-wide pairwise shared analysis. Significant P-values support evidence of genetic sharing based on the correlation of significa
association findings reported by GWAS for each pair of diseases. ARTICLE
NATURE COMMUNICATIONS | DOI: 10.1038/ncomms944 ARTICLE 0.25
0.20
0.15
0.10
0.05
0.00
Prevalence of subjects w/ D1 among Pts with D2
Disease 1 – Disease 2
CD-UC
UC-CD
CVID-JIA
CEL-PS
T1D-CEL
CD-SPA
PS-UC
JIA-CEL
PS-T1D
PS-CD
UC-SPA
SLE-T1D
CEL-T1D
CD-PS
UC-PS
CVID-CD
SLE-UC
UC-T1D
CEL-SLE
CEL-CD
SLE-PS
CD-T1D
T1D-JIA
JIA-T1D
CVID-SLE
T1D-PS
CEL-UC
T1D-UC
CD-CEL
SLE-CD
CEl-SPA
T1D-CD
UC-CEL
PS-CEL
PS-SLE
S:E-CEL
CD-SLE
UC-SLE
T1D-SLE
CEL-JIA
Pairwise correlation or H2 (per disease)
1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0
–0.2
–0.4
–0.6
CVID-JIA EPI-JIA
EPI-UC PSOR-T1D PS-UC
SLE-CD
SPA-CD
T1D-CD
UC-CD
D1 SNP-h2
Autosomal rG
exMHC rG
D2 SNP-h2
0
0.88
1.77
2.65
3.54
4.42
CD
JIA
T1D
UC
CVID
SLE
CEL
PS
SPA
THY
SPA
PSOR
CEL
SLE
CVID
UC
T1D
JIA
a
b
c
Figure 2 | Prevalence of AI disease co-morbidities and estimates of genetic correlation (co-heritability) across pAIDs. (a) Observed prevalence of pAI
comorbidity observed in Caucasian populations in Europe and North America as curated from large-scale cohort studies. For each pairwise combinatio
(for example, Disease 1–Disease 2), the rate (y axis) indicates the percentage of patients with Disease 2 who have also been diagnosed with Disease
Literature data used and references can be found in Supplementary Table 9. (b) Bivariate estimates of genetic correlation (pairwise co-heritability) acros
pAIDs. The heritability (SNP-h2) for the first and second disease are shown for each pAID pair (blue and green bars, respectively) along with the genet
correlation (rG) for the pair estimated based on total autosomal common genetic variants (orange) and based on autosomal variants excluding the MH
(red). Displayed are those pairs for which the rG estimates reached nominal significance (Po0.05). P-values are based on restricted maximum likelihoo
ratio test. Error bars represent standard error. (c) Genetic sharing using the genome-wide pairwise sharing statistic (GPS). Correlation plot of the P-value
obtained from the genome-wide pairwise shared analysis. Discussion
k To our knowledge, this is the most comprehensive assessment of
heritability and disease prediction using genome-wide dense
genotyping data across multiple pAIDs. The results show that
SNP-h2 estimates were significantly higher for the pAID cohorts
as compared with those obtained for the non-immune-mediated
disease EPI (Fig. 1a and Supplementary Tables 1 and 2). Among
the pAIDs examined where the SNP-h2 estimates were at least
nominally significant (Po0.05), T1D and JIA were the most
highly heritable (Fig. 1b). These results are in keeping with the
SNP-h2 estimates reported for T1D and Rheumatoid Factor
Positive (RF þ ), Rheumatoid Arthritis (RA) in adults, using the
Wellcome Trust Case Control Consortium data sets17,26,31. Considerably weaker SNP-h2 estimates were observed for UC
and
CD,
consistent
with
previous
reports
in
adults32
(Supplementary Fig. 1A). Although the sample size of CD was
several fold greater than those of T1D and UC, and twice that for
JIA, the SNP-h2 estimates are lower in CD, suggesting that
environmental factors play a much larger role in CD disease
aetiology (Fig. 1d). This finding is in keeping with studies
demonstrating a key role for the gut microbiome and faecal flora
in disease-onset and severity in the IBDs11,33,34. A number of previous epidemiological and genetic studies
have suggested a significant degree of shared risk across AI
diseases44–47. There are a number of reasons why our results may
differ from these reports. In such population-based studies,
observed
sharing
of
risk
in
the
population
is
inevitably
confounded
by
common
environmental
factors
or
gene–
environment interactions, neither of which would be parsed out
from purely epidemiological observations. In addition, it can be
challenging to perform these comparisons in heterogeneous
populations
because
they
may
be
composed
of
different
underlying genetic backgrounds, and genetic ancestry is known
to dramatically affect the risk for many AI diseases (for example,
greater risk of CEL and JIA in Caucasians)4,20. g
Although there are several prior large-scale analyses of genetic
sharing among AI diseases using GWAS data, these are based on
somewhat different analytical approaches or study methodology
than those employed here. A notable example comes from
Cotsapas et al., who derived a Cross-Phenotype Meta-Analysis
test statistic that powerfully combines multiple independent AI
data sets to analyse the likelihood that a SNP is shared across
disease phenotypes. They applied this test statistic to the 140 top
genetic risk variants reported previously by GWAS across seven
AI diseases47. ARTICLE Shown are the mean (orange) and maximum (blue) areas under the curve
(AUC) achieved in the validation set as obtained for each disease in the
ten-fold cross-validation analysis. The mean and maxima refer to the best
AUC’s when testing a range of P-value thresholds from which to pick SNPs
in training the linear SVM. SPA, spondyloarthropathy. Figure 3 | Disease prediction using a support vector machine model. Figure 3 | Disease prediction using a support vector machine model. Shown are the mean (orange) and maximum (blue) areas under the curve
(AUC) achieved in the validation set as obtained for each disease in the
ten-fold cross-validation analysis. The mean and maxima refer to the best
AUC’s when testing a range of P-value thresholds from which to pick SNPs
in training the linear SVM. SPA, spondyloarthropathy. g
Although
estimates
for
JIA
and
T1D
are
higher
than
SNP-h2 estimates reported previously, our estimates for RA and
T1D are more consistent, although still falling short of, than those
reported by population estimates from twin-based or familial
studies (Supplementary Table 2). That these SNP-h2 based
estimates are in general still falling behind estimates made from
epidemiological studies illustrates the ‘missing heritability’ phe-
nomenon. Disparities between POP-h2 and SNP-h2 estimates may
be at least partially attributable to inflation of population-based
estimates in the presence of ascertainment-bias and/or insufficient
adjustment for confounding effects. The latter tends to occur if
there are significant non-additive or shared environmental factors
that contribute to phenotypic variation36,43. the extended MHC was entirely removed from the analysis across
any of the pAID pairs, making it unlikely that the sharing of
common HLA alleles could significantly account for the degree of
co-heritability observed (Fig. 2b). ARTICLE ARTICLE 0.5
0.55
0.6
0.65
0.7
0.75
0.8
0.85
0.9
0.95
1JIA
CD
UC
T1D
CEL
PS
SLE
SPA
THY
CVID
Avg
Max
Figure 3 | Disease prediction using a support vector machine model. Shown are the mean (orange) and maximum (blue) areas under the curve
(AUC) achieved in the validation set as obtained for each disease in the
ten-fold cross-validation analysis. The mean and maxima refer to the best
AUC’s when testing a range of P-value thresholds from which to pick SNPs
in training the linear SVM. SPA, spondyloarthropathy. 0.5
0.55
0.6
0.65
0.7
0.75
0.8
0.85
0.9
0.95
1JIA
CD
UC
T1D
CEL
PS
SLE
SPA
THY
CVID
Avg
Max RF patients across different study cohorts, as recent analyses
suggest that RF þ RA may be ‘distinct’ from RF forms of RA
in terms of genetic aetiology37. Moreover, the subphenotype of
JIA that is most similar to RF þ RA (i.e. RF þ JIA) made up only
a small component of our JIA cohort (4.9%). Thus, the high
estimated heritability observed in JIA suggests that despite the
heterogeneous clinical findings, there may be a strongly shared
genetic component contributing to a common aetiology. 2 UC g
p
g
gy
We observed relatively low SNP-h2 estimates for SLE (Fig. 1b). Although these estimates are lower than those reported by So
et al.,38,39 they are higher than the POP-h2 reported based on
sibling-recurrence40. These observations are consistent with strong
environmental and epigenetic components to SLE liability41,42. We
included in our analysis a non-immune-mediated disease, early-
onset EPI, as a comparator cohort. As expected, the SNP-h2
estimates on the liability scale, albeit non-zero, was relatively low
compared with any of the AI diseases. That we observed slightly
higher heritability estimates across our paediatric cohorts than
previously reported in adults is also in keeping with the notion that
paediatric-onset diseases have been noted previously to reflect
disease aetiologies with a stronger genetic component29 and less
confounding
due
to
reduced
timespan
of
environmental
exposure(s). Adult or late-onset AI diseases can be associated
with environmental precipitating factors such as viral infections or
drug exposures, which have been implicated in a range of AI
diseases including T1D, CEL and SLE3,42. PS PS Figure 3 | Disease prediction using a support vector machine model. & 2015 Macmillan Publishers Limited. All rights reserved. NATURE COMMUNICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | www.nature.com/naturecommunications NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 We also
noted a positive rG between common variable immunodeficiency
disorder (CVID) and JIA (rG ¼ þ 0.34), although it was more
modest (PBLo0.01). While we did observe a marginally positive
rG for CD and T1D consistent with results from published
GWAS metanalysis30, although it did not reach significance at a
liberal Bonferroni threshold (rG ¼ þ 0.096; PBL ¼ 0.17). Of note,
we did not observe a significant reduction in rG estimates when Estimation
of
pairwise
co-heritability
across
pAIDs. To
investigate diseases with shared underlying genetic risk factors,
we assessed the genetic correlation (rG) for each pair of pAIDs NATURE COMMUNICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | www.nature.com/naturecommunications
& 2015 Macmillan Publishers Limited. All rights reserved. 4 NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 NATURE COMMUNICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | www.nature.com/naturecommunications
& 2015 Macmillan Publishers Limited. All rights reserved. |
|
/
|
/
& 2015 Macmillan Publishers Limited. All rights reserved. ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 heterogeneity
these
studies
often
combine
summary
data
obtained from independent case–control study cohorts accrued
and genotyped across North America and Europe using different
genotyping platforms and QC/analysis steps, requiring post-hoc
statistical adjustments for heterogeneity, genetic variation and the
use of SNP proxies. Although single-institution study designs can
have limited applicability, in our study, using a common shared
control accrued in the same institution and genotyped on the
same platform does limit the effect of inter-study heterogeneity in
our analysis. predictive SNPs without significantly impacting maximum or
mean AUC achieved, suggesting that the SVM model was robust
to this parameter (Fig. 3 and Supplementary Table 11). In
comparison, we obtained fairly modest AUCs for CD, UC and
CVID (AUCmax40.7, AUCmean40.65). These are in keeping
with our expectation that genetic prediction should rest on
underlying genetic heritability and confirms the value of SNP
heritability analysis. Indeed, the above observations are perhaps not surprising,
given recent findings that support a strong contribution for
environmental factors in disease susceptibility. For example, host-
microbial interactions have been implicated in the pathogenesis
of IBD and RA11,54. Furthermore, in CVID, it is well-established
that although genetic risk factors play a role in disease risk, there
is significant within-disease heterogeneity in terms of aetiology. Patients with CVID are often diagnosed in late adolescence,
suggesting that environmental risk factors play a greater role. Likewise, most cases of paediatric-onset IBD also have a post-
pubescent age of onset. This is in contrast to T1D, JIA or CEL,
which are commonly diagnosed by or before the age of 12 years,
although some degree of variability is observed. This is consistent
with the correlations noted above, in that the three diseases with
more moderate SNP-h2 estimates were also less predictable. As expected, we found that variations across the extended
MHC strongly contributed to both heritability estimates and
disease risk predictability in T1D and CEL, and more modestly in
UC and JIA. The contribution of the extended MHC to total
phenotypic variance explained correlated with the strength of the
strongest association signal within the extended MHC. However,
as recent reports have shown, this method for estimating h2 is
sensitive to the variation in linkage-disequilibrium (LD) across
the genome18,31. We therefore examined the effect of LD on the
SNP-h2 estimates by comparing the results with those obtained
using non-correlated SNP markers (Supplementary Fig. 2B; see
Methods for details). ARTICLE As anticipated, the effect of the pruning is
mostly attributable to the strong role of the MHC in the
heritability of these diseases, as pruning had little effect on the
heritability estimates once the extended MHC was removed. Thus, the number and degree of LD for the input SNPs used for
calculating h2 can be important for diseases where the MHC plays
a major role, consistent with previous studies31,48,49. 2 p
Among the three largest cohorts, namely JIA, UC and CD, CD
was by far the largest. However, the heritability estimated for CD
in our data set was the lowest of the three. As we know from prior
studies that disease prediction is a function of heritability, sample
size and the number of causal variants, we might expect the
accuracy of disease prediction for CD to be relatively poor. This is
exactly what we observed. In contrast, we had somewhat limited
sample sizes for SPA, PS and CEL cohorts, and we caution against
the interpretation of the high heritability estimates observed for
PS. Another limitation of the present study is that we have not
considered the role of rare, or potentially de novo, variants in the
overall estimates of genetic heritability. As more sequencing data
using either whole-exome or whole-genome approaches become
available, future studies will help address this question. j
p
The SNP-h2 for T1D was most strongly affected by the removal
of the extended MHC, emphasizing the importance of MHC
polymorphisms in T1D pathogenesis. In addition, the estimates
for SPA and CEL both fell significantly when markers across the
MHC were excluded from further analysis. The relatively limited
contribution of the genetic polymorphisms across the MHC to
heritability in IBD was consistent with prior GWAS results,
as the MHC SNP (rs1626392, Po2.27 10 7) most significantly
associated with CD did not reach genome-wide significance,
defined as Po5 10 8 (Supplementary Table 8). Aside from the
MHC, recent work has examined the degree to which functional
or coding loci, for example, DNAse I Hypersensitivity Sites50,
contribute
to
disease
heritability. Such
studies,
currently
underway, will help delineate biological functions and connect
genetic associations with mechanistic roles of such functional
variants. Discussion
k Although there is no doubt that findings from this
study are informative, the targeted candidate approach has clear
limitations and only summary statistics were available. Another
concern, which is not unique to the study by Cotsapas et al., but a
concern in most large GWAS meta-analyses, is inter-study y
As noted, the SNP-h2 observed for JIA was high despite the
known heterogeneous nature of this disease, including seven
distinct JIA subtypes35. Little is known about the heritability of
JIA as it is fairly uncommon. However, in RA, the more common
JIA counterpart in adults, a range of SNP-h2 estimates has been
reported17,26,31,36. Some of the heterogeneity in SNP-h2 estimates
for RA may be attributable to the different ratios of RF þ vs 5 ARTICLE A unique opportunity provided by our cohort was the ability
to quantify pairwise pAID genetic correlations as numerous
epidemiological analyses have shown that subsets of pAIDs
co-cluster in families or exhibit high rates of comorbidity55–57
(Table 3). As pAID co-heritability has not been systematically
examined using genome-wide SNP data, we aimed to identify
pAIDs showing significantly positive rG (that are consequently
co-heritable) versus diseases that are either genetically unrelated
or
negatively
correlated
(and
are
consequently
‘mutually-
protective’). We calculated the rG for each pAID pair and
between each of the nine diseases and EPI. This latter analysis
provides a ‘control’ or contextual baseline, akin to the inclusion of A still unrealized, but much anticipated goal of personalized
medicine is to utilize genomic data to accurately predict disease
risk26,51–53. We found that for the three pAIDs (T1D, JIA and
CEL) that were most predictable, a range of P-value thresholds
(Po1 10 6 and Po1 10 8) could be used to identify the Table 3 | pAID joint heritabilities or genetic correlation (rG) reaching nominal significance. pAID pair
rG (auto)
s.e. Pval
rG (exMHC)
s.e. P_nominal
P_adj
CVID-JIA
0.343
0.127
1.22E–03
0.354
0.142
2.47E–03
2.23E–02
EPI-JIA
0.150
0.079
2.95E–02
0.142
0.085
4.87E–02
0.44
EPI-UC
0.197
0.103
2.77E–02
0.248
0.108
1.06E–02
0.10
PS-T1D
0.241
0.139
3.29E–02
0.282
0.167
3.74E–02
0.34
PS-UC
0.316
0.169
2.31E–02
0.289
0.171
3.76E–02
0.34
SLE-CD
0.266
0.120
8.25E–03
0.255
0.121
1.15E–02
0.10
SPA-CD
0.215
0.138
4.64E–02
0.235
0.156
4.67E–02
0.42
T1D-CD
0.096
0.053
3.45E–02
0.142
0.064
1.33E–02
0.12
UC-CD
0.659
0.069
o1.00E–04
0.674
0.072
o1.00E–04
9.00E–04
Auto, autosomal; CD, crohn’s disease; CEL, celiac disease; CVID, common variable immunodeficiency disorder; EPI, epilepsy; JIA, juvenile idiopathic arthritis; NA, not applicable; pAID, paediatric
autoimmune disease; PS, psoriasis; exMHC, MHC excluded; SLE, systemic lupus erythematosus; SPA, spondyloarthropathy; T1D, type 1 diabetes; UC, ulcerative colitis. P-values (P) are based on results from the restricted maximum likelihood estimate (likelihood ratio test). P_adj is made using a Bonferonni-adjustment for nine pairwise tests for each disease. Table 3 | pAID joint heritabilities or genetic correlation (rG) reaching nominal significance. Auto, autosomal; CD, crohn’s disease; CEL, celiac disease; CVID, common variable immunodeficiency disorder; EPI, epilepsy; JIA, juvenile idiopathic arthritis; NA, not applicable; pAID, paediatric
autoimmune disease; PS, psoriasis; exMHC, MHC excluded; SLE, systemic lupus erythematosus; SPA, spondyloarthropathy; T1D, type 1 diabetes; UC, ulcerative colitis. ARTICLE Taken together, we report genome-wide SNP genotype-derived
heritability estimates and genetic correlations of disease liability
across pairs of nine investigated pAIDs using common and low-
frequency genetic variants. We contextualized these findings
alongside a comprehensive review of available literature and
epidemiological data sets, illustrate a method for quantifying
genetic risk factor sharing across pAIDs and provide considera-
tions for how such genetic data can aid in disease prediction. We observed that SNP-h2 estimates in pediatric AI diseases tend to
be greater in magnitude when compared to SNP-h2 reported
previously based on GWAS data from studies of adult AI disease
cohorts, particularly for T1D, UC, JIA/RA. Moreover, we also
observed that the ‘co-heritability’ across pAIDs was minimally
attributable to shared MHC variations. While genomic screening in
the general population on a large scale is not currently feasible, or
of high utility (given the low disease prevalence and consequently,
limited positive predictive value as well as the limitations in
interpretability), our analysis suggests that there is a high
heritability and disease predictability across the pAIDs. Future
studies in larger sample sizes and in adult cohorts will be helpful in
validating these results and developing new and improved methods
for genome-based disease prediction and for the development of
novel biomarkers that can be used to predict pAID risk. The balance of the paediatric AI disease subjects’ (SPA, PS, CEL and SLE)
samples were accrued by our biorepository at the CHOP, which includes over
60,000 paediatric patients recruited and enrolled by the Center for Applied
Genomics at CHOP. These individuals were ascertained for having a confirmed
diagnosis of SPA, PS, CEL and SLE in the age range of 1–17 years during time of
diagnosis and were required to fulfill clinical criteria for these respective disorders,
as confirmed by a specialist. Only cases that upon EMR search were confirmed to
have at least two or more in-person visits, at least one of which is with the specified
ICD9 diagnosis code(s) were pursued for clinical confirmation (see Supplementary
Table 12 for ICD-9 inclusion and exclusion codes). We used ICD9 codes previously
identified and utilized for PheWAS or EMR-based GWAS59,60 and agreed upon by
board-certified physicians. ARTICLE Disease diagnosis was based on these clinical criteria, rather than any
laboratory tests. The JIA cohort was recruited in the United States of America, Australia and
Norway and comprised of a total of 1,123 patients with onset of arthritis at
o16 years of age. JIA diagnosis and JIA subtype were determined according to the
International League of Associations for Rheumatology revised criteria35 and
confirmed using the JIA Calculator software66 (http://www.jra-research.org/
JIAcalc/), an algorithm-based tool adapted from the International League of
Associations for Rheumatology criteria. Before standard QC procedures and
exclusion of non-European ancestry, the JIA cohort was comprised of 464 case
subjects from Texas Scottish Rite Hospital for Children (Dallas, Texas, USA)
and the Children’s Mercy Hospitals and Clinics (Kansas City, Missouri, USA)
of self-reported European ancestry; 196 subjects from the CHOP; 221 subjects
from the Murdoch Childrens Research Institute (Royal Children’s Hospital,
Melbourne, Australia) and 504 subjects from the Oslo University Hospital
(Oslo, Norway). Somewhat unexpectedly, we observed a negative marginal rG
across several pAID pairs, including SLE-CD, SPA-CD, PS-UC
and PS-T1D. Although none of these was significant following a
Bonferroni correction, in each of the negatively correlated pAID
pairs, one of the two diseases is considered a ‘classic autoimmune’
(that is, SLE, UC, and T1D), whereas the other pAID in the pair
(that is, CD and PS) has been noted to have a strong
‘inflammatory’ component. The CVID study population consisted of 223 patients from the Mount Sinai
School of Medicine (New York City, New York, USA); 76 patients from the
University of Oxford (London, England); 47 patients from the CHOP and 27
patients from the University of South Florida (Tampa, Florida). The diagnosis in
each case was validated against the ESID/PAGID diagnostic criteria, as previously
described67. Although the diagnosis of CVID is most commonly made in young
adults (aged 20–40 years), all of the CHOP and University of South Florida cases
had paediatric age of onset disease, whereas the majority of the cases from the
Mount Sinai School of Medicine and Oxford had onset in young adulthood. We note that as the number of individuals with adult-onset CVID disease is so
small (less than 5% of all cases presented), and all ten diseases have paediatric age
of onset disease, we have elected to refer to the cohort material as pAIDs. ARTICLE ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 CD as a ‘null comparator’ phenotype by the Psychiatric Genomics
Consortium20. We observed a strongly and moderately positive
rG between two pAID pairs, namely UC-CD and JIA-CVID. sample size from which we excluded non-qualified samples/genotypes that did not
pass QC criteria required for inclusion in the genetic analysis (for example, because
of relatedness or poor genotyping rate; see details below). p
g
yp
g
)
The IBD cohort comprised 2,796 individuals aged 2–17 years of European
ancestry with biopsy-proven disease, including 1,931 with CD and 865 with UC,
excluding all patients with unclassified type (IBD-U). Affected individuals were
recruited from multiple centres from four geographically discrete countries and
diagnosed before their nineteenth birthday according to the standard IBD
diagnostic criteria, as previously reported3,29. Although the MHC made major contributions to the disease-
specific heritability, we found no evidence that variations across
the MHC significantly contributed to the pAID co-heritability for
any of the investigated disease pairs (Fig. 2b). For the pAID pairs
with significantly positive rG’s (UC-CD and JIA-CVID), we did
not observe a significant reduction in rG estimates when the SNPs
within the MHC were removed from the analysis, making it
unlikely that genetic sharing of MHC haplotypes can explain the
genetic correlation observed among pAIDs in this data set
(Fig. 2b). In addition, that the UC-CD and JIA-CVID pairs were
the two with the largest positive rG is also consistent with results
we obtained using an independent genome-wide pairwise sharing
metric for genetic correlation, in which we considered all
genome-wide SNP markers except those within the extended
MHC locus (Fig. 2c, see Methods for details). Although it may
appear to be surprising given the known association with the
MHC across all pAIDs, these results are in keeping with our
finding that the most significant MHC association signals
identified for each pAID was disease-specific and did not overlap
across the nine pAIDs (Supplementary Table 5). The T1D cohort consisted of 1,120 cases from nuclear family trios (one affected
child and two parents), including 267 independent Canadian T1D cases collected
in paediatric diabetes clinics in Montreal, Toronto, Ottawa and Winnipeg (Canada)
and 203 T1D cases recruited at CHOP since September 2006. All patients were
Caucasians by self-report and ranged in age between 3 and 17 years, with 7.9 years
being the median age at onset. All patients have been treated with insulin since
diagnosis. ARTICLE p y
Age- and gender-matched control subjects, including the EPI cohort of both
generalized and focal idiopathic EPI (ICD-9 345.9 and 345.4, respectively),
were identified from the CHOP-CAG biobank and ascertained by exclusion
of any patient with any ICD-9 codes for disorders of autoimmunity or
immunodeficiency58 (http://eicd9.com/). Research Ethics Boards at the CHOP and
each of the collaborating centre, including: the Mount Sinai School of Medicine,
University of Oxford, University of South Florida, the Children’s Mercy Hospitals
and Clinics, Texas Scottish Rite Hospital for Children, Murdoch Children’s
Research Institute, Oslo University Hospital, Cincinnati Children’s Hospital
Medical Center, McGill University, RCCS ‘Casa Sollievo della Sofferenza’,
University of Toronto, University of Edinburgh, Emory University, University of
Naples ‘Federico II’, Cedars Sinai Medical Center, Yorkhill Hospital for Sick
Children, University of Miami Miller School of Medicine, Careggi University
Hospital, University of Utah School of Medicine and Primary Children’s Medical
Center, approved this study. ARTICLE P-values (P) are based on results from the restricted maximum likelihood estimate (likelihood ratio test). P_adj is made using a Bonferonni-adjustment for nine pairwise tests for each disease. NATURE COMMUNICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | www.nature.com/naturecommunications
& 2015 Macmillan Publishers Limited. All rights reserved. 6 & 2015 Macmillan Publishers Limited. All rights reserved. Y1; . . . ; Yn are the Z scores for D2 across n SNPs ð2Þ The test statistic, g, used to detect genetic sharing between two diseases is ð3Þ g ¼ max
1jn min xj
; yj
; which is the maximum of the pairwise minima of the signals across all of the
n SNPs. The rationale is that if SNP j is associated with both D1 and D2, the
magnitudes of both Xj and Yj should be large. The more shared SNPs there are, the
greater the likelihood that the maximum of the pairwise minimal values will be
large. Under the null hypothesis that any genetic sharing is due only to chance,
g should be relatively small. We can obtain the P-value of this statistic by
permuting the labels of the Z-scores relative to each other in order to simulate the
null hypothesis. In fact, these P-values can be calculated analytically using a
hypergeometric distribution, and no actual permutation is needed. Note that no
significance threshold is required. This test was performed for all 45 pairwise pAID
combinations (hence, the reported P-values are Bonferroni-adjusted for 45 inde-
pendent tests). Web-based access to all novel data included in this manuscript is available
through our website at http://www.caglab.org. Population stratification correction. The final cohort, following all above-noted
QC, included a total of 4,956 pAID cases inclusive of 9 pAIDs and 27,451
population-matched controls, as well as a cohort consisting of 819 cases of
paediatric-onset EPI. To avoid confounding, we assigned individuals fitting the
diagnosis criteria for two or more pAIDs to the smaller disease cohort by sample
size. No individual was included twice. To ensure that the markers tested across the
cohorts were consistent, we included only SNPs that passed all QC criteria (461,301
SNPs). The filtered SNPs were tested in cases and controls for association with
disease and used for the estimation of the genetic relationship matrix (see below). We used a logistic regression equation to estimate ORs/betas, 95% confidence
intervals and P-values for trend, using additive coding for genotypes (0,1,2 minor
alleles). We adjusted for gender and population stratification by including the
binary gender and the first ten PCs (GCTA) from the PCA calculated from a set of
100,000 pruned SNPs as covariates in the logistic regression analyses69. From the
results of the association testing, we determined the genomic inflation per disease-
common control cohort. References 1. Anaya, J.-M., Go´mez, L. & Castiblanco, J. Is there a common genetic basis for
autoimmune diseases? Clin. Dev. Immunol. 13, 185–195 (2006). 2. Rojas-Villarraga, A., Amaya-Amaya, J., Rodriguez-Rodriguez, A.,
Mantilla, R. D. & Anaya, J.-M. Introducing polyautoimmunity: secondary
autoimmune diseases no longer exist. Autoimmune Dis. 2012, 254319 (201 We applied the previously described linear mixed model method for estimating
whole-genome SNP-based heritability using both common and low-frequency
variants, which is implemented in the software GCTA. We estimated the
genetic variance associated with genome-wide SNPs on the observed scale
(SNP-heritability or SNP-h2)70, conditioning on the top 20 ancestry PCs derived
from a pruned set of B100,000 independent SNPs across the same data set (that is,
PLINK --indep-pairwise 50 10 0.2) obtained also using GCTA. As our phenotypes
are dichotomous traits, we subsequently transformed these results to the liability
scale based on approximately observed disease prevalence at our centre for each
trait (Table 1). Note that the total control sample size utilized varied slightly as we
optimized our analysis to maximize the retained sample size when conservatively
removing distantly related individuals during QC. As we excluded rare variants
(MAFo0.01), these variants are therefore not included in the heritability estimates
attributable to genetic variation. g
3. Lettre, G. & Rioux, J. D. Autoimmune diseases: insights from genome-wide
association studies. Hum. Mol. Genet 17, R116–R121 (2008). 4. Nunes, T., Fiorino, G., Danese, S. & Sans, M. Familial aggregation in
inflammatory bowel disease: is it genes or environment? World J. Gastroenterol. 17, 2715–2722 (2011). 5. Cooper, J. D. et al. Seven newly identified loci for autoimmune thyroid disease. Hum. Mol. Genet. 21, 5202–5208 (2012). 6. Tsoi, L. C. et al. Identification of 15 new psoriasis susceptibility loci highlights
the role of innate immunity. Nat. Genet. 44, 1341 (2012). 7. Hinks, A. et al. Dense genotyping of immune-related disease regions identifies
14 new susceptibility loci for juvenile idiopathic arthritis. Nat. Genet. 45,
664–669 (2013). 8. L, J. et al. Dense fine-mapping study identifies new susceptibility loci for
primary biliary cirrhosis. Nat. Genet. 44, 1137 (2012). Joint heritability across pAID pairwise combinations. We estimated the genetic
correlation in disease risk for each of other pAID pairs using a bivariate linear
mixed model, as described previously17. For each pairwise analysis, the pooled
control samples passing QC were randomly allocated to the two diseases evenly
and the top 20 PCs were again included as covariates. Y1; . . . ; Yn are the Z scores for D2 across n SNPs All disease-specific, case–control GWAS had lGC values at
or below 1.04 with the exception of CD (1.09), consistent with that previously
reported for this data set29. Final counts from each pAID cohort, included controls
and genomic inflation calculated from median w2 association test statistics are
reported in Table 1. Disease prediction using a linear SVM. Given that we observed relatively high
rates of heritability across many of pAIDs, we sought to evaluate the utility of
genome-wide SNP data in predicting pAID disease liability, using a previously
described SVM pipeline that can be applied to GWAS results for a dichotomous
trait52. We identified SNPs to be used as predictors based on the strength of association
with a given disease in a training set, testing graded P-value thresholds
(Po1 10 5, 1 10 6, 1 10 7, 1 10 8, 1 10 9) for selecting SNP
predictors, where the P-value is derived from the case–control association testing
using samples in the training data set. We used each set of SNPs passing the tested
threshold to then train the linear SVM model. We then validated the SVM model by testing the accuracy of disease liability
predictions for each of the nine pAIDs in the remaining independent sample set. We reported the prediction performance as the mean and maximum AUC
achieved in both the training and validation sets (Fig. 3 and Supplementary
Table 11). Estimation of the variance components for each pAID. Only individuals and
SNPs that passed all QC metrics were used to estimate the variance components
for the ten diseases (nine pAIDs and one non-pAID condition EPI). For disease-
specific analysis, the common set of controls were used for each case–control
analysis cohort, after excluding individuals who are relatives up to within the
5th degree. The genetic relationship between individuals was estimated using
(i) all autosomal SNPs, (ii) all autosomal SNPs excluding the extended MHC
(chr6:26.5–34 Mb) and (iii) SNPs only found on the X-chromosome (ChrX). ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 because of the presence of samples with non-matching platforms, this is not ideal
given (i) added risk of artefacts and (ii) consequent variations in coverage
(genotyping density) across the genome. Without adding significant additional
information, this can result in biased heritability estimates unless careful
corrections are made to apply additional down-sampling/weighting of more
densely imputed regions31,48,49. As over 90% of the markers on the two arrays are
shared, whole-genome imputation was not necessary and we utilized only the set of
directly overlapping genotyped SNPs in the analysis (B500,000). each of two diseases; these SNPs are the genetic risk factors shared by that pair of
pAIDs. Most existing tests require choosing a significance threshold to determine
which SNPs are associated with which disease, but it is unknown how best to
choose this threshold. Our method is threshold-free and requires no tuning
parameters. Specifically, for any two diseases, we converted the P-values for all
SNPs in the genome into Z-scores, such that for example: X1; . . . ; Xn are the Z scores for D1 across n SNPs;
ð1Þ
Y1; . . . ; Yn are the Z scores for D2 across n SNPs:
ð2Þ
The test statistic, g, used to detect genetic sharing between two diseases is
g ¼ max
1jn min xj
; yj
;
ð3Þ ð1Þ After extracting the overlapping SNPs from the two platforms, SNPs with a low
genotyping rate o95%, low MAF (o0.01) or significantly departing from the
expected Hardy–Weinberg equilibrium (Po0.01) were excluded. Samples with low
average genotyping call rate (o95%) or determined to be of outliers of European
ancestry by PCA (Any of the top ten principal components (PCs) 46.0 standard
deviations as reported by SMARTPCA/EIGENSTRAT68) were removed. In
addition, one of each pair of distantly related individuals, as determined by
Identify-by-State analysis (40.05), was excluded, such that the largest sample size
would be retained in the final cohort. Methods
St d Study population. Information regarding the patient cohorts have been published
previously and are summarized briefly below. Written informed consent was obtained from all subjects (or their legal
guardians). Genomic DNA extraction and sample QC before and following
genotyping were performed using standard methods61. To minimize confounding
because of population stratification, we focused on only individuals of European
ancestry, as determined by both self-reported ancestry and principle component
analysis, PCA) in the present study (see below and Supplementary Fig. 4). Cases and controls were either directly ascertained as described in prior
studies29,53,58–65 or obtained from de-identified samples and associated electronic
medical records (EMRs) residing in the genomics biorepository at the Children’s
Hospital of Philadelphia. EMR searches were conducted using previously described
algorithms58,59 based on phenotype mapping established using PheWAS ICD-9
code mapping tables53,58,60 in consultation with qualified physician specialists for
each disease cohort. All DNA samples were assessed for QC and genotyped on the
Illumina HumanHap550 or HumanHap610 platforms at the Center for Applied
Genomics (CAG) at the Children’s Hospital of Philadelphia (CHOP, Philadelphia,
Pennsylvania, USA). Note that the patient counts below refer to the total recruited Genotyping and QC. All samples were genotyped at the CAG on the Human-
Hap550 or 610 BeadChip arrays. Although some published analyses using GWAS
data to derive heritability estimates have applied whole-genome imputation 7 NATURE COMMUNICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | www.nature.com/naturecommunications NATURE COMMUNICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | w NATURE COMMUNICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | www.nature.com/naturecommunications
& 2015 Macmillan Publishers Limited. All rights reserved. & 2015 Macmillan Publishers Limited. All rights reserved. NATURE COMMUNICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | www.nature.com/naturecommunications References H., Yang, J., Goddard, M. E., Visscher, P. M. & Wray, N. R. Estimation
of pleiotropy between complex diseases using single-nucleotide polymorphism-
derived genomic relationships and restricted maximum likelihood. Bioinformatics 28, 2540–2542 (2012). 46. EC, S., Cooper, G. S., Bynum, M. L. K. & Somers, E. C. Recent insights
in the epidemiology of autoimmune diseases: improved prevalence
estimates and understanding of clustering of diseases. J. Autoimmun. 33,
197–207 (2009). Bioinformatics 28, 2540–2542 (2012). 18. Lee, S. H., Wray, N. R., Goddard, M. E. & Visscher, P. M. Estimating missing
heritability for disease from genome-wide association studies. Am. J. Hum. Genet. 88, 294–305 (2011). 47. Cotsapas, C. et al. Pervasive sharing of genetic effects in autoimmune disease. PLoS Genet. 7, e1002254 (2011). 48. Speed, D. et al. SNP-based heritability analysis with dense data. Am. J. Hum. Genet. 93, 1155–1157 (2013). 19. Yang, J. et al. Common SNPs explain a large proportion of the heritability for
human height. Nat. Genet. 42, 565–569 (2010). 49. Lee, S. H. et al. Estimation of SNP heritability from dense genotype data. Am. J. Hum. Genet. 93, 1151–1155 (2013). 20. Lee, S. H. et al. Genetic relationship between five psychiatric disorders
estimated from genome-wide SNPs. Nat. Genet. 45, 984–994 (2013). 50. Gusev, A. et al. Partitioning heritability of regulatory and cell-type-specific
variants across 11 common diseases. Am. J. Hum. Genet. 95, 535–552
(2014). g
21. Jia, X. et al. Imputing amino acid polymorphisms in human leukocyte antigens. PLoS ONE 8, e64683 ( (2013). 22. Schaffner, S. F. The X chromosome in population genetics. Nat. Rev. Genet. 5,
43–51 (2004). 51. Kang, J., Kugathasan, S., Georges, M., Zhao, H. & Cho, J. H. Improved risk
prediction for Crohn’s disease with a multi-locus approach. Hum. Mol. Genet
20, 2435–2442 (2011). 23. Gottipati, S., Arbiza, L., Siepel, A., Clark, A. G. & Keinan, A. Analyses of
X-linked and autosomal genetic variation in population-scale whole genome
sequencing. Nat. Genet. 43, 741–743 (2011). 52. Mittag, F. et al. Use of support vector machines for disease risk prediction in
genome-wide association studies: Concerns and opportunities. Hum. Mutat. 33, 1708–1718 (2012). q
g
24. Daetwyler, H. D., Villanueva, B. & Woolliams, J. A. Accuracy of predicting the
genetic risk of disease using a genome-wide approach. PLoS ONE 3, e3395
(2008). 53. Liao, K. P. et al. Electronic medical records for discovery research in
rheumatoid arthritis. Arthritis Care Res. (Hoboken) 62, 1120–1127 (2010). 25. References Loci on 20q13 and 21q22 are associated with
pediatric-onset inflammatory bowel disease. Nat. Genet. 40, 1211–1215 (2008). 33. Cho, I. & Blaser, M. J. The human microbiome: at the interface of health and
disease. Nat. Rev. Genet. 13, 260–270 (2012). 63. Orange, J. S. et al. Genome-wide association identifies diverse causes of
common variable immunodeficiency. J. Allergy Clin. Immunol. 127, 1360–1367
e6 (2011). 34. Ma´rquez, A. et al. Specific association of a CLEC16A/KIAA0350 polymorphism
with NOD2/CARD15(-) Crohn’s disease patients. Eur. J. Hum. Genet. 17,
1304–1308 (2009). 64. Behrens, E. M. et al. Association of the TRAF1-C5 locus on chromosome 9 with
juvenile idiopathic arthritis. Arthritis Rheum. 58, 2206–2207 (2008). 65. Grant, S. F. et al. Association of the BANK 1 R61H variant with systemic lupus
erythematosus in Americans of European and African ancestry. Appl. Clin. Genet. 2, 1–5 (2009). 35. Petty, R. E. et al. International League of Associations for Rheumatology
classification of juvenile idiopathic arthritis: second revision, Edmonton, 2001. J. Rheumatol. 31, 390–392 (2004). 66. Behrens, E. M. et al. Evaluation of the presentation of systemic onset juvenile
rheumatoid arthritis: data from the Pennsylvania Systemic Onset Juvenile
Arthritis Registry (PASOJAR). J. Rheumatol. 35, 343–348 (2008). 36. Zaitlen, N. et al. Using extended genealogy to estimate components of
heritability for 23 quantitative and dichotomous traits. PLoS Genet. 9, e1003520
(2013). 67. Conley, M. E., Notarangelo, L. D. & Etzioni, A. Diagnostic criteria for primary
immunodeficiencies. Representing PAGID (Pan-American Group for
Immunodeficiency) and ESID (European Society for Immunodeficiencies). Clin. Immunol. 93, 190–197 (1999). 37. Han, B. et al. Fine mapping seronegative and seropositive rheumatoid arthritis
to shared and distinct HLA alleles by adjusting for the effects of heterogeneity. Am. J. Hum. Genet. 94, 522–532 (2014). 38. So, H.-C., Gui, A. H. S., Cherny, S. S. & Sham, P. C. Evaluating the heritability
explained by known susceptibility variants: a survey of ten complex diseases. Genet. Epidemiol. 35, 310–317 (2011). 68. Price, A. L. et al. Principal components analysis corrects for stratification in
genome-wide association studies. Nat. Genet. 38, 904 (2006). 69. Purcell, S. et al. PLINK: a tool set for whole-genome association and
population-based linkage analyses. Am. J. Hum. Genet. 81, 559–575 (2007). p
39. So, H.-C., Yip, B. H. K. & Sham, P. C. Estimating the total number of
susceptibility variants underlying complex diseases from genome-wide
association studies. PLoS ONE 5, e13898 (2010). 70. NASU, Y. et al. References By jointly analysing a pair of
cohorts, these results estimate both the SNP-h2 of liability to both diseases and an
estimate of the SNP-genetic correlation between these liabilities. We determined
the significance of the rG using a likelihood ratio test by fixing the genetic
correlation at zero17. Significantly positive (or negative) rG’s should reflect a shared
(or disparate) genetic background, as a positive (negative) rG means that the
correlation in the genetic variance components are higher (lower) between case
subjects than between the case subjects and the respective control cohorts. 9. Liu, J. Z. et al. Dense genotyping of immune-related disease regions identifies
nine new risk loci for primary sclerosing cholangitis. Nat. Genet. 45, 670–675
(2013). 10. Eyre, S. et al. High-density genetic mapping identifies new susceptibility loci for
rheumatoid arthritis. Nat. Genet. 44, 1336 (2012). 11. DP, M. et al. Host-microbe interactions have shaped the genetic architecture of
inflammatory bowel disease. Nature 491, 119 (2012). 12. Beecham, A. H. et al. Analysis of immune-related loci identifies 48 new
susceptibility variants for multiple sclerosis. Nat. Genet. 45, 1353–1360 (201 13. Zhernakova, A. et al. Detecting shared pathogenesis from the shared genetics of
immune-related diseases. Nat. Rev. Genet. 10, 43–55 (2009). 14. Parkes, M., Cortes, A., van Heel, D. A. & Brown, M. A. Genetic insights into
common pathways and complex relationships among immune-mediated
diseases. Nat. Rev. Genet. 14, 661–673 (2013). Genome-wide pairwise sharing analysis. We applied a novel test to detect the
presence of SNPs anywhere in the genome that are simultaneously associated with NATURE COMMUNICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | www.nature.com/naturecommunications 8 & 2015 Macmillan Publishers Limited. All rights reserved. ARTICLE ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/ncomms9442 44. Rzhetsky, A., Wajngurt, D., Park, N. & Zheng, T. Probing genetic overlap
among complex human phenotypes. Proc. Natl Acad. Sci. USA 104,
11694–11699 (2007). 15. Visscher, P. M., Hill, W. G. & Wray, N. R. Heritability in the genomics
era--concepts and misconceptions. Nat. Rev. Genet. 9, 255–266 (2008). 16. Yang, J., Lee, S. H., Goddard, M. E. & Visscher, P. M. GCTA: a tool
for genome-wide complex trait analysis. Am. J. Hum. Genet. 88, 76–82
(2011). g
for genome-wide complex trait analysis. Am. J. Hum. Genet. 88, 76–82
(2011). 45. Eaton, W. W., Rose, N. R., Kalaydjian, A., Pedersen, M. G. & Mortensen, P. B. Epidemiology of autoimmune diseases in Denmark. J. Autoimmun. 29, 1–9
(2007). 17. Lee, S. References Trichostatin A, a histone deacetylase inhibitor, suppresses
synovial inflammation and subsequent cartilage destruction in a collagen
antibody-induced arthritis mouse model. Osteoarthr. Cartil. 16, 723–732
(2008). 40. Harley, J. B. et al. Genome-wide association scan in women with systemic lupus
erythematosus identifies susceptibility variants in ITGAM, PXK, KIAA1542
and other loci. Nat. Genet. 40, 204–210 (2008). 41. Kamen, D. L. Environmental influences on systemic lupus erythematosus
expression. Rheum. Dis. Clin. North Am 40, 401–412 vii (2014). References Lee, S. H. & Wray, N. R. Novel genetic analysis for case-control genome-wide
association studies: quantification of power and genomic prediction accuracy. PLoS ONE 8, e71494 (2013). 54. Scher, J. U. et al. Expansion of intestinal Prevotella copri correlates with
enhanced susceptibility to arthritis. Elife 2, e01202 (2013). 55. Ramos, P. S. et al. A comprehensive analysis of shared loci between systemic
lupus erythematosus (SLE) and sixteen autoimmune diseases reveals limited
genetic overlap. PLoS Genet. 7, e1002406 (2011). 26. Speed, D. & Balding, D. J. MultiBLUP: improved SNP-based prediction for
complex traits. Genome Res 24, 1550–1557 (2014). 27. Franke, A. et al. Genome-wide meta-analysis increases to 71 the number
of confirmed Crohn’s disease susceptibility loci. Nat. Genet. 42, 1118–1125
(2010). 56. Tait, K. F. et al. Clustering of autoimmune disease in parents of siblings from
the Type 1 diabetes Warren repository. Diabet. Med. 21, 358–362 (2004). 57. Lin, J.-P. et al. Familial clustering of rheumatoid arthritis with other
autoimmune diseases. Hum. Genet. 103, 475–482 (1998). 28. Barrett, J. C. et al. Genome-wide association study of ulcerative colitis identifies
three new susceptibility loci, including the HNF4A region. Nat. Genet. 41,
1330–1334 (2009). 58. Denny, J. C. et al. PheWAS: demonstrating the feasibility of a phenome-wide
scan to discover gene-disease associations. Bioinformatics 26, 1205–1210 (2010). 29. Imielinski, M. et al. Common variants at five new loci associated with
early-onset inflammatory bowel disease. Nat. Genet. 41, 1335–1340 (2009). 59. Ritchie, M. D. et al. Robust replication of genotype-phenotype associations
across multiple diseases in an electronic medical record. Am. J. Hum. Genet. 86,
560–572 (2010). 30. Wang, K. et al. Comparative genetic analysis of inflammatory bowel disease and
type 1 diabetes implicates multiple loci with opposite effects. Hum Mol Genet
19, 2059–2067 (2010). 60. Liao, K. P. et al. Associations of autoantibodies, autoimmune risk alleles, and
clinical diagnoses from the electronic medical records in rheumatoid arthritis
cases and non-rheumatoid arthritis controls. Arthritis Rheum. 65, 571–581
(2013). 31. Speed, D., Hemani, G., Johnson, M. R. & Balding, D. J. Improved
heritability estimation from genome-wide SNPs. Am. J. Hum. Genet. 91,
1011–1021 (2012). 61. Hakonarson, H. et al. A genome-wide association study identifies KIAA0350 as
a type 1 diabetes gene. Nature 448, 591 (2007). 32. Chen, G.-B. et al. Estimation and partitioning of (co)heritability of
inflammatory bowel disease from GWAS and immunochip data. Hum. Mol. Genet 23, 4710–4720 (2014). 62. Kugathasan, S. et al. Competing financial interests: The authors declare no competing financial. Reprints and permission information is available online at http://npg.nature.com/
reprintsandpermissions/ Supplementary Information accompanies this paper at http://www.nature.com/
naturecommunications Competing financial interests: The authors declare no competing financial. Acknowledgements 42. Mok, C. C. & Lau, C. S. Pathogenesis of systemic lupus erythematosus. J. Clin
Pathol. 56, 481–490 (2003). 42. Mok, C. C. & Lau, C. S. Pathogenesis of systemic lupus erythematosus. J. Clin. Pathol. 56, 481–490 (2003). We thank the patients and their families for their participation in the genotyping studies
and in the Biobank Repository at the Center for Applied Genomics. We are also thankful
for the contributions of the Italian IBD Group, including Cucchiara S (Roma), Lionetti P
(Firenze), Barabino G (Genova), de Angelis GL (Parma), Guariso G (Padova), Catassi C 43. Manolio, T. A. et al. Finding the missing heritability of complex diseases. Nature 461, 747–753 (2009). NATURE COMMUNICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | www.nature.com/naturecommunications 9 & 2015 Macmillan Publishers Limited. All rights reserved. NATURE COMMUNICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | www.nature.com/naturecommunications
& 2015 Macmillan Publishers Limited. All rights reserved. Additional information (Ancona), Lombardi G (Pescara), Staiano AM (Napoli), De Venuto D (Bari), Romano C
(Messina), D’inca` R (Padova), Vecchi M (Milano), Andriulli A and Bossa F (S. Giovanni
Rotondo). Y.R.L. is supported by the Paul and Daisy Soros Fellowship for New Americans
and an NIH F30 Individual NRSA Training Grant (1F30AR066486). This study was
supported by Institutional Development Funds from the Children’s Hospital of Philadel-
phia, and by DP3DK085708, RC1AR058606, U01HG006830, the Crohn’s and Colitis
Foundation, the Juvenile Diabetes Research Foundation and a grant from the LRI to E.L.P. Supplementary Information accompanies this paper at http://www.nature.com/
naturecommunications Author contributions How to cite this article: Li, Y. R. et al. Genetic sharing and heritability of paediatric
age of onset autoimmune diseases. Nat. Commun. 6:8442 doi: 10.1038/ncomms9442
(2015). Y.R.L. and H.H. were leading contributors in the design, analysis and writing of
this study. D.J.A., M.M. and L.S. contributed to data collection and literature review. B.F., Ø.F., L.A.D., S.D.T., M.L.B., S.L.G., A.L., E.P., E.R., C.S., A.S., E.M., M.S.S.,
B.A.L., M.P., R.K.R., D.C.W., H.C., C.C.-R., J.S.O., E.M.B., K.E.S., S.K., A.M.G.,
J.S., T.F., C.P., R.N.B. and J.A.E. contributed samples and phenotypes. F.D.M., K.A.T., H.Q., R.M.C., C.E.K., F.W. and J.S. provided assistance with
samples genotyping, and data processing. J.K., S.D.Z., J.P.B., J.L. and H.L. contributed to,
advised and supervised statistical analysis. E.T.L.P., J.A.E. and B.J.K. assisted in
composing and revising the manuscript. All authors read, edited and approved of
the manuscript. This work is licensed under a Creative Commons Attribution 4.0
International License. The images or other third party material in this
article are included in the article’s Creative Commons license, unless indicated otherwise
in the credit line; if the material is not included under the Creative Commons license,
users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ 10 NATURE COMMUNICATIONS | 6:8442 | DOI: 10.1038/ncomms9442 | www.nature.com/naturecommunications
& 2015 Macmillan Publishers Limited. All rights reserved.
|
https://openalex.org/W4393020650
|
https://ojs.unimal.ac.id/jpes/article/download/13295/5514
|
Indonesian
| null |
Pelatihan Wirausaha Perempuan Dalam Proses Pengolahan Ikan Menjadi Fried Meatballs Di Desa Cot Keumuneng Aceh Utara
|
Jurnal Pengabdian Ekonomi dan Sosial
| 2,023
|
cc-by-sa
| 1,519
|
Jurnal Pengabdian Ekonomi dan Sosial
Vol 2 No 2 Oktober 2023 Jurnal Pengabdian Ekonomi dan Sosial
Vol 2 No 2 Oktober 2023 ISSN:2962-5831(Online - Elektronik) ABSTRAK Inovasi merupakan kemampuan seseorang individu atau sekelompok orang untuk
menghasilkan sesuatu yang baru seperti gagasan dan karya nyata yang relatif berbeda dengan
produk yang sebelumnya telah ada. Proses inovasi menjadi saluran untuk menciptakan dan
memberikan nilai di sebuah usaha. Fried Meatballs merupakan bentuk inovasi dari olahan
dari daging ikan sebagai bentuk peningkatan nilai ekonomi masyarakat. Fried Meatballs
atau lebih dikenal dengan bakso goreng merupakan olahan bahan baku ikan dengan bahan
pengisi berpati atau tepung tapioka dan bumbu-bumbu dalam bentuk bulat. peluang bisnis
ini di masyarakat memiliki potensi berbentuk makanan yang ringan jika di pasarkan
memberikan nilai ekonomis. Kata Kunci: pelatihan, ikan, friend meatballs PELATIHAN WIRAUSAHA PEREMPUAN DALAM PROSES
PENGOLAHAN IKAN MENJADI FRIED MEATBALLS
DI DESA COT KEUMUNENG ACEH UTARA zailaton1 , agustinawati2 zailaton1 , agustinawati2 1,2 Program studi kewirausahaan, Fakultas Ekonomi dan Bisnis Universitas Malikussaleh
*Email korespondesi: agustinawati@unimal.ac.id 1,2 Program studi kewirausahaan, Fakultas Ekonomi dan Bisnis Universitas Malikussaleh
*Email korespondesi: agustinawati@unimal.ac.id PENDAHULUAN Inovasi dimaknai sebagai suatu bentuk dari perbauran terhadap suatu produk dalam
negeri dengan tingkatan pada Ekonomi yang kuat. Oleh karena itu dalam kontek Inovasi
tentunya juga membutuhkan keberhasilan suatu tindakan dalam upaya wirausaha untuk
meningkatkan eksistensi bisnis. Inovasi merupakan kemampuan seseorang individu atau
sekelompok orang untuk menghasilkan sesuatu yang baru seperti gagasan dan karya nyata
yang relatif berbeda dengan produk yang sebelumnya telah ada (Aidhi et al,. 2023). Inovasi
juga dapat berpengaruh terhadap kinerja pengembangan produk dalam sebuah usaha
(Rachmasari & Suprapti, 2022). Proses inovasi menjadi saluran untuk menciptakan dan
memberikan nilai di sebuah usaha. Maka inovasi bisnis dijadikan sebagai mediasi
penghubung antara orientasi kewirausahaan terhadap kinerja pengembangan produk baru
yang saling berpengaruh antara satu dan lainya. Di sisi lain untuk memaksimalkan pengembangan produk diperlukan suatu orientasi
kewirausahaan menjadi kunci dalam meningkatkan kinerja bisnis sehingga prinsipnya
produk tersebut dapat menarik perhatian masyarakat. Orientasi kewirausahaan dijadikan
dalam konstruksi tingkat usaha, strategi keseluruhan organisasi dan modal bisnis. Melihat
peluang tersebut dengan perkembangan dan pembangunan bentuk inovasi semakin
meningkatnya aktivitas masyarakat Fried Meatballs merupakan bentuk inovasi dari olahan
dari daging ikan sebagai bentuk peningkatan nilai ekonomi masyarakat. Fried Meatballs
atau lebih dikenal dengan Bakso goreng disajikan mengikuti kemajuan dari inovasi produk-
produk makanan yang disesuaikan pada kebutuhan pasar. Namun tidak hanya pada bentuk
nilai jual saja, bakso goreng tentunya juga memiliki manfaat di antaranya (1) lauk makan;
(2) Snack; dan (3) cemilan ketika santai. Bakso goreng ini merupakan suatu inovasi yang
mampu memenuhi kebutuhan masyarakat. Dengan kata lain bakso goreng di inovasika Pelatihan Wirausaha Perempuan (Zailaton, dkk.) | 19 Vol.2 No.2 Vol.2 No.2 Jurnal Pengabdian Ekonomi dan Sosial
Vol 2 No 2 Oktober 2023 ISSN:2962-5831(Online - Elektronik) dengan tujuan untuk pemberdayaan Wirausaha Perempuan di Desa Cot keumuneng Aceh
Utara. Pelaksanan
kegiatan
pengabdian
kepada
masyarakat
bertujuan
untuk
mensejahterakan Wirausaha Perempuan, namun masih terbatas sarana dan prasarana seperti;
(1) keterbatasan pengetahuan masyarakat terkait dengan penunjang usaha; dan (2)
Keterbatasan kader kewirausahaan masyarakat. Berdasarkan permasalahan tersebut kami
mahasiswa prodi kewirausahaan menggagas kegiatan pengabdian kepada masyarakat
Pelatihan Wirausaha Perempuan Dalam Proses Pengolahan Ikan Menjadi Fried Meatballs
Di Desa Cot Keumuneng Aceh Utara. METODE Pelaksanan kegiatan pemberdayaan kepada masyarakat ini dilaksanakan di Desa Cot
Keumuneng, dengan peserta pelatihan adalah masyarakat sebanyak 20 orang. kegiatan
pemberdayaan kepada masyarakat ini Di Desa Cot Keumuneng ini dibentuk atas dasar
kebutuhan mitra. Namun untuk sampai pada tujuan yang dimaksud metode pelaksanaan
pengabdian ini dirancang sebagai berikut: A. Perencanaan
Pada tanggal 13 September 2022 dilakukan observasi dengan pihak mitra Desa Cot
Keumuneng, sehingga dilakukan perumusan permasalahan. Selanjutnya, dilakukan
penyusunan pelaksanaan program berdasarkan pada kebutuhan mitra. B. Persiapan
Pelaksanaan yang dilakukan dengan persiapan yang strategis berupa pengumpulan alat,
bahan dan materi saat sosialisasi di antara nya. a) Materi sosialisasi disampaikan terkait dengan bentuk semangat pembuatan
i
i d i b k B. Persiapan
Pelaksanaan yang dilakukan dengan persiapan yang strategis berupa pengumpulan alat,
bahan dan materi saat sosialisasi di antara nya. ) M
i
i li
i di
ik
k i
d
b
k
b Pelaksanaan yang dilakukan dengan persiapan yang strategis berupa pengumpulan alat a) Materi sosialisasi disampaikan terkait dengan bentuk semangat pembuatan
inovasi dari bakso goreng b) Pembuatan bahan Bakso goreng yang dikemas dalam bentuk inovasi c) Pembuatan kemasan bakso goreng sebagai brend produk yang akan dipasarkan
C. Pelaksanan
Pelaksanaan ini dilakukan program ini dilaksanakan selama 3 (tiga) kali yang dimulai
dari kunjungan observasi, pelaksanaan sampai dengan evaluasi kegiatan dengan
masyarakat Cot Keumuneng elaksanaan ini dilakukan program ini dilaksanakan selama 3 (tiga) kali yang dimulai
ari kunjungan observasi, pelaksanaan sampai dengan evaluasi kegiatan dengan
masyarakat Cot Keumuneng a) Melakukan sosialisasi dengan pendampingan kepada masyarakat b) Melakukan pemotongan bahan olahan dalam inovasi bakso goreng c) Praktik pengolahan dan penggorengan bakso goreng d) Selanjutnya pelatihan pengemasan bakso goreng dalam bentuk brand yang akan
dipasarkan Pelatihan Wirausaha Perempuan (Zailaton, dkk.) | 20 D. Evaluasi D. Evaluasi
Kegiatan pengabdian kepada masyarakat ini dilaksanakan pada 3 kali pertemuan dengan
masyarakat yang terlibat dalam mengevaluasi kegiatan ini yaitu tim pelaksana serta
kader yang dibentuk. Selanjutnya dilakukan analisis terhadap pengetahuan dan
keberlanjutan program yang nanti nya dilakukan oleh masyarakat. (a)
(b)
(c)
Gambar 1 (a). Pengenalan bahan ; (b) Penggorengan ; (c) Meatballs yang dikemas (c)
Meatballs yang dikemas (b) (a) (a) (b) (c) Gambar 1 (a). Pengenalan bahan ; (b) Penggorengan ; (c) Meatballs yang dikemas Pelatihan Wirausaha Perempuan (Zailaton, dkk.) | 20 Vol.2 No.2 Vol.2 No.2 Jurnal Pengabdian Ekonomi dan Sosial
Vol 2 No 2 Oktober 2023 ISSN:2962-5831(Online - Elektronik) HASIL, PEMBAHASAN, DAN DAMPAK (12pt) Secara umum makanan menjadi sumber nutrisi bahkan energi yang berasal dari
tumbuhan dan hewan bagi makhluk hidup. Setiap makanan tentunya dapat membuka
peluang bisnis bagi masyarakat yang dilalui proses inovasi (Hertanius, 2019). Jika ditinjau
dari landasan peluang bisnis di masyarakat memiliki potensi berbentuk makanan yang ringan
jika di pasarkan memberikan nilai ekonomis (Amanda at al., 2022). Inovasi dari bakso goreng merupakan olahan bahan baku ikan dengan bahan pengisi
berpati atau tepung tapioka dan bumbu-bumbu dalam bentuk bulat (Agustin, 2020). Pengabdian ini dilaksanakan selama tiga (3) kali secara langsung. Kelompok sasaran dalam
kegiatan pengabdian ini adalah masyarakat dengan jumlah 20 (dua puluh) orang. Kelompok
sasaran ini dipilih atas dasar karena pertimbangan efektivitas kegiatan serta pembentukan
kader guna meneruskan inovasi kepada masyarakat di lingkungan sosial. Hasil yang dicapai dalam pelatihan ini adalah hasil rancangan inovasi bakso goreng
sesuai dengan kebutuhan masyarakat. Rancangan dan metode yang digunakan melalui
pendekatan kepada masyarakat guna menjawab batasan pengetahuan masyarakat. Adapun
dampak kegiatan pengabdian kepada masyarakat yang disesuaikan dengan permasalahan
dan kebutuhan mitra dapat dilihat pada tabel 1. Tabel 1. Dampak Perubahan Pada Kelompok Sasaran
No
Permasalahan dan
Kebutuhan Mitra
Intervensi
Hasil
1
Keterbatasan
pengetahuan
masyarakat
terkait
dengan
nilai
penunjang
kewirausahaan
Sosialisasi
dengan
menggunakan
praktik
Mendapatkan
penambahan
pemahaman
dan
pengetahuan
masyarakat
terkait
dengan
penunjang
kewirausahaan
yaitu
inovasi bakso goreng
2
Tidak
ada
penggerak
dari
program
pemberdayaan
masyarakat sebagai
penunjang
Inovasi
Kewirausahaan
Praktik
pengolahan
bakso
goreng
melalui Inovasi
sebagai bentuk
peningkatan
pendapatan
wirausaha
perempun
Terbentuknya
kader
kewirausahan
perempuan
dalam
bentuk pemberdayaan
usaha
wirausaha
lainnyadi
Desa
Cot
Keumuneng. Tabel 1. Dampak Perubahan Pada Kelompok Sasaran KESIMPULAN Hasil yang diperoleh dari kegiatan ini adalah wirausaha perempuan mendapatkan ilmu
pengetahuan dalam mengelola atau produksi hingga pemasaran yang diterapkan dalam
proses kehidupan sehari-hari. Pendekatan ini dapat diadopsi dan dikembangkan lebih lanjut
kepada masyarakat dalam peningkatan nilai ekonomis lainya. Kedua implementasi dan
keberlanjutan dari program pemberdayaan masyarakat ini dapat menjalin komitmen kader
inovasi dari masyarakat. Selanjutnya juga konsep dari inovasi kewirausahaan turut dirasakan
oleh masyarakat dalam konteks memenuhi kebutuhan dasar bahkan meningkat ekonomi
perempuan masyarakat desa Cot Keumuneng. Pelatihan Wirausaha Perempuan (Zailaton, dkk.) | 21 Vol.2 No.2 Jurnal Pengabdian Ekonomi dan Sosial
Vol 2 No 2 Oktober 2023 ISSN:2962-5831(Online - Elektronik) Pelatihan Wirausaha Perempuan (Zailaton, dkk.) | 22 UCAPAN TERIMAKASIH Kegiatan pengabdian kepada masyarakat ini tidak dapat dilaksanakan tanpa
dukungan banyak pihak. oleh karenanya, kami mengucapkan terimakasih kepada Dosen
Pendamping lapangan atas dukungan secara administrasi dan substansi dalam pelaksanaan
program, Kepada Desa Cot Keumuneng sebagai mitra yang telah mendukung dan ikut
berpartisipasi dalam kegiatan ini dan semua pihak yang tidak dapat disebutkan satu persatu. DAFTAR PUSTAKA Agustin, L. (2020). Produksi Dan Pemasaran Bakso Goreng “Basreng “Ikan
Tongkol (Doctoral Dissertation, Politeknik Negeri Jember). Akbarani, T. N. (2023). Pengaruh Customer Satisfaction, Usability Dan Trust Terhadap
Kemudahan Dan Kenyamanan Bertransaksi Di Bank Syariah Tanggamus Dalam
Perspektif Ekonomi Islam (Doctoral Dissertation, Ui n Raden Intan Lampung). Al Aidhi, A., Harahap, M. A. K., Rukmana, A. Y., & Bakri, A. A. (2023). Peningkatan Daya
Saing Ekonomi Melalui Peranan Inovasi. Jurnal Multidisiplin West Science, 2(02),
118-134. Amanda, F. R., Tarmizi, A., & Habibah, G. W. I. (2022). Nilai-Nilai Ekonomi Syariah Pada
Usaha Rumahan Dodol Ketan Di Desa Teluk Rendah Pasar Kecamatan Tebo Ilir
Kabupaten Tebo (Doctoral Dissertation, Uin Sulthan Thaha Saifuddin Jambi). Firmansyah, M. R. (2022). Strategi Pemasaran Syariah Pada Bakso Yummy Sebagai Salah
Satu Cemilan Yang Sehat Dan Bergizi (Doctoral Dissertation, Uin Fatmawati
Sukarno Bengkulu). Hertanius, F. (2019). Proses Inovasi Munculnya Ide Dan Kreativitas Dalam Terciptanya
Kampung Pelangi Wonosari Semarang (Doctoral Dissertation). Rachmasari, A. D., & Suprapti, N. W. S. (2022). Peran Inovasi Produk Dalam Memediasi
Pengaruh Orientasi Pasar Terhadap Kinerja Bisnis Ukm Kedai Kopi (Doctoral
Dissertation, Udayana University Vol.2 No.2
|
https://openalex.org/W3198750743
|
https://www.ijhpm.com/article_4112_4e47085d1bfd86c767ac63f52696b43a.pdf
|
English
| null |
Public-Private Partnerships With Unhealthy Commodity Industries: Are They Undermining Real Progress in Non-Communicable Disease Prevention? Competing Frames in Global Health Governance: An Analysis of Stakeholder Influence on the Political Declaration on Non-communicable Diseases
|
International journal of health policy and management
| 2,021
|
cc-by
| 2,492
|
Article History:
Received: 11 June 2021
Accepted: 25 August 2021
ePublished: 30 August 2021 https://ijhpm.com https://ijhpm.com https://ijhpm.com
Int J Health Policy Manag 2022, 11(7), 1212–1214 https://ijhpm.com
Int J Health Policy Manag 2022, 11(7), 1212–1214 doi 10.34172/ijhpm.2021.118 Public-Private Partnerships With Unhealthy Commodity
Industries: Are They Undermining Real Progress in Non-
communicable Disease Prevention?
Comment on “Competing Frames in Global Health Governance: An Analysis of Stakehold
Influence on the Political Declaration on Non-communicable Diseases” Public-Private Partnerships With Unhealthy Commodity
Industries: Are They Undermining Real Progress in Non-
communicable Disease Prevention? Comment on “Competing Frames in Global Health Governance: An Analysis of Stakeholder
Influence on the Political Declaration on Non-communicable Diseases” Abstract Article History:
Received: 11 June 2021
Accepted: 25 August 2021
ePublished: 30 August 2021 Public-private partnerships (PPPs) and whole-of-society approaches are increasingly common in public health
promotion and non-communicable disease (NCD) prevention, despite a lack of evidence in favour of their effectiveness
in improving health outcomes. While PPPs may have advantages, they also give industry actors more influence over
the design and implementation of public health strategies and interventions. Partnering with unhealthy commodity
industries in particular – including the alcohol and ultra-processed food and beverages industries – can pose significant
risks to public health due to these industries’ deep-rooted conflicts of interest. In this commentary, I reiterate Suzuki
and colleagues’ message about the importance of assessing and managing conflicts of interest before engaging with
non-state actors through PPPs or other forms of engagement. Keywords: Public-Private Partnerships, Non-Communicable Diseases, Global Health Governance, Conflicts of
Interest Copyright: © 2022 The Author(s); Published by Kerman University of Medical Sciences. This is an open-access article
distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/
by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is
properly cited. p
p
y
Citation: Rinaldi C. Public-private partnerships with unhealthy commodity industries: are they undermining real
progress in non-communicable disease prevention?: Comment on “Competing frames in global health governance:
an analysis of stakeholder influence on the political declaration on non-communicable diseases.” Int J Health Policy
Manag. 2022;11(7):1212–1214. doi:10.34172/ijhpm.2021.118 *Correspondence to:
Chiara Rinaldi
Email:
chiara.rinaldi@lshtm.ac.uk *Correspondence to:
Chiara Rinaldi
Email:
chiara.rinaldi@lshtm.ac.uk Commentary Commentary Department of Health Services Research and Policy, London School of Hygiene and Tropical Medicine, London, U *Correspondence to:
Chiara Rinaldi
Email:
chiara.rinaldi@lshtm.ac.uk Background groups, and inequalities in whose framing was adopted by
the UN in the final Declaration. Notably, the issues raised
by non-governmental organizations (NGOs) and low- and
middle-income countries (LMICs) were less likely to be
adopted than those raised by industry stakeholders and high-
income countries. As Suzuki and colleagues’ analysis shows,
unhealthy commodity industries like the alcohol and ultra-
processed food and beverages industries are often treated as
‘partners’ in NCD governance despite the growing evidence
that the framing and strategies they adopt interfere with the
implementation of evidence-based public health measures.2
In line with previous analyses, the authors question whether
and how unhealthy commodity industries should be formally
involved in global NCD policy deliberations. The commercial determinants of health, or “factors that
influence health which stem from the profit motive” (p. 687),1
are a growing area of interest in public health research. The
impact of unhealthy commodity industries – including the
tobacco, alcohol and ultra-processed food and beverages
industries – on non-communicable diseases (NCDs) is of
particular concern due to the ways in which those actors have
been able to influence policy-making processes. Unhealthy
commodity industries have a clear conflict of interest in NCD
prevention because of their financial interests in producing,
marketing and selling products that contribute to the burden
of NCDs.3 However, while the conflicts of interest of the
tobacco industry are generally well understood and managed,
Suzuki and colleagues’ analysis highlights the need to identify
and evaluate other industries that have significant and often
irreconcilable conflicts of interest in the prevention of NCDs.2 Public-Private Partnerships and Whole-of-Society Approaches
One of the main themes that emerged from Suzuki and
colleagues’ analysis is the promotion of public-private
partnerships (PPPs) and ‘whole-of-society’ approaches,
predominantly by private sector actors. In recent years, PPPs
have increasingly been adopted as a tool to address common
risk factors for the development of NCDs – such as unhealthy
diets, physical inactivity and obesity – and other health- In their analysis of stakeholder influence on the United
Nations (UN) Political Declaration on NCDs, Suzuki et al
document the extent to which the Declaration reflects the
policy positions advocated by different public, private and
third sector stakeholders during the consultation process.2
They identified divergent framing between stakeholder Rinaldi industry to reduce the ‘harmful use of alcohol’ and eliminate
‘the marketing, advertising and sale of alcoholic products to
minors’” (p. Effectiveness of PPPs in Preventing NCDs Effectiveness of PPPs in Preventing NCDs PPPs are often conceptualised as political practices through
which corporations can contribute to addressing the negative
externalities of their products or practices, while at the same
time promoting their preferred regulatory environment by
participating in decision-making processes.9 Both creating
a more favourable image for themselves – often as part
of corporate social responsibility – and having increased
influence over strategies and interventions to address NCDs
can be a reason for unhealthy commodity industries to
partner with governments or international organisations,
and to promote such partnerships in consultation processes. Benefits of PPPs for the public sector can include increased
access to (financial) resources and expertise. What is interesting to note about Suzuki and colleagues’
findings is that when there was no direct threat to their own
business, industry stakeholder did not necessarily oppose
more wide-ranging regulatory interventions.2 For example,
an association of pharmaceutical industries supported the
taxation of sugar-sweetened beverages. This further highlights
the importance of carefully assessing and managing conflicts
of interest when engaging in PPPs and acknowledging
that some industry stakeholders are not in the position to
contribute to decision-making process in a way that promotes
evidence-based public health. It is therefore concerning that
the UN Declaration on NCDs does not make any attempt to
distinguish between the roles of different non-state actors
and their (conflicting) interests despite thirty statements of
support in the consultation process. In Suzuki and colleagues’
words, this could indeed be “the worst possible combination
(promotion of PPPs without the management of conflict of
interest) from the perspective of many NGOs and public
health advocates” (p. 10).2 However, there is no clear evidence in favour of the
effectiveness of PPPs in public health promotion to date.3,10,11
A recent systematic review found that partnerships addressing
NCD risk factors, which have high potential for conflicts
of interest, are more likely to be reported as unsuccessful
in reaching their goals compared to PPPs in communicable
disease control.10 Moreover, the authors of the review expressed
concern about the amount of ‘non-independent’ or industry-
funded studies on PPPs, which were in turn more likely to
be supportive of the evaluated partnerships. Effectiveness of PPPs in Preventing NCDs An evaluation
of the UK Public Health Responsibility Deal, a PPP based on
voluntary pledges from the alcohol and food and beverage
industries, suggests that beyond just being ineffective, taking
a PPP approach could delay the implementation of evidence-
based public health regulation that is less favourable from
a business perspective (eg, restrictions to the availability of
products and fiscal policies).11 International Journal of Health Policy and Management, 2022, 11(7), 1212–1214
1213 Background 9).2 While this appears positive, it does require
comment. The alcohol industry tends to disproportionally
focus on the harms of their products on the heaviest drinkers
or particularly vulnerable groups (eg, minors) while rejecting
harms in the wider population,8,12 despite evidence that the
harms of alcohol start from a low level of consumption.13
This framing is also commonly used by the tobacco and
ultra-processed food and beverage industries, and explains
why their commitments and pledges have a similar, narrow
focus.3 Limited support for interventions targeted at specific
populations does not represent a comprehensive public
health approach to NCD prevention – particularly given that
the interventions that are favoured by unhealthy commodity
industries (eg, education and awareness raising) tend to
have a weaker evidence base.14 The increased influence of
industry stakeholders in PPPs can lead to the adoption of a
narrow individual-oriented framing of NCD prevention at
the expense of an approach guided by public health values
and evidence.11 related challenges.4 PPPs represent a shift in governance
for health, in which responsibility is shared between public
bodies and other societal stakeholders.5 ‘Multi-stakeholder’ or
‘whole-of-society’ approaches are promoted by the UN, for
example through Sustainable Development Goal 17 which
includes the target to “encourage and promote effective public,
public-private and civil society partnerships.”6 While ‘whole-
of-society’ approaches are broader than PPPs and often aim
to engage civil society stakeholders, there is evidence that
the term is used by representatives of the alcohol and food
industry to justify and promote industry involvement in
policymaking, and to oppose attempts to limit involvement
due to conflicts of interest.7,8 The use of the terms ‘whole-
of-society’ and ‘whole-of-government’ together without
explicit differentiation, as seen in the UN Declaration on
NCDs, further adds to this ambiguity about the meaning of
different terms that indicate the engagement of sectors and
stakeholders beyond health.2 Funding Funding
No funding was received for this commentary. CR is supported by a National
Institute for Health Research (NIHR) School for Public Health Research (SPHR)
Pre-doctoral Fellowship. 14. Anderson P, Chisholm D, Fuhr DC. Effectiveness and cost-effectiveness
of policies and programmes to reduce the harm caused by alcohol. Lancet. 2009;373(9682):2234-2246. doi:10.1016/s0140-6736(09)60744-3 15. Maani N, Van Schalkwyk MC, Petticrew M, Ralston R, Collin J. The new
WHO Foundation - global health deserves better. BMJ Glob Health. 2021;6(2):e004950. doi:10.1136/bmjgh-2021-004950 International Journal of Health Policy and Management, 2022, 11(7), 1212–1214
1214 1.
West R, Marteau T. Commentary on Casswell (2013): the commercial
determinants of health. Addiction. 2013;108(4):686-687. doi:10.1111/ Ethical issues
Not applicable. Ethical issues
Not applicable. 11. Knai C, Petticrew M, Douglas N, et al. The public health responsibility deal:
using a systems-level analysis to understand the lack of impact on alcohol,
food, physical activity, and workplace health sub-systems. Int J Environ
Res Public Health. 2018;15(12):2895. doi:10.3390/ijerph15122895 Not applicable. Competing interests
Author declares that she has no competing interests. Competing interests
Author declares that she has no competing interests. 12. McCambridge J, Mialon M, Hawkins B. Alcohol industry involvement in
policymaking: a systematic review. Addiction. 2018;113(9):1571-1584. doi:10.1111/add.14216 Author’s contribution
CR is the single author of the paper. Author’s contribution
CR is the single author of the paper. 13. Burton R, Sheron N. No level of alcohol consumption improves health. Lancet. 2018;392(10152):987-988. doi:10.1016/s0140-6736(18)31571-x Conclusion
h Given the current evidence base, the increasing shift towards
PPPs could represent a concern for public health, specifically
when it comes to the prevention and control of NCDs. The
promotion of ‘multi-stakeholder’ and ‘whole-of-society’
approaches by the UN, with strong backing of unhealthy
commodity industries, should therefore be done with caution. It is important to note that most evidence on PPPs focuses
on formal partnerships established for the implementation
of specific public health interventions with a shared goal. However, there are different ways in which partnerships
can be organised, including more informal participation-
based approaches with ‘engagement’ or ‘dialogue’ between
sectors. Evidence on such partnerships remains limited. Regardless, we also need to carefully consider these other,
less overt ways in which unhealthy commodity industries
can influence strategies and approaches to NCD prevention, The Promotion of Industry Framing of NCD Prevention As could be expected, Suzuki and colleagues’ analysis shows
that industry stakeholders are opposed to regulation that could
directly harm their business (eg, taxation on sugar-sweetened
beverages).2 On the other hand, they note that “there was
consensus from all constituencies including the alcohol Rinaldi including through financial donations and through their
status as ‘partners’ in policy deliberations and consultations. The World Health Organization Foundation has recently
sparked controversy for accepting donations from harmful
commodity industries, seemingly undermining Framework
of Engagement with Non-State Actors principles which aim
to protect WHO’s work from conflicts of interest.15 Suzuki
and colleagues’ findings show that in the consultation process
on the UN Declaration on NCDs, contestation was associated
with a smaller likelihood of an issue being addressed and
included in the Declaration with clear language. In this light,
involving those with strong vested interests in policy outcomes
in deliberations risks overshadowing important issues raised
by NGOs and LMICs, giving industry stakeholders and
high-income countries with strong ties to those industries
disproportionate power. This is particularly concerning given
the high and rising burden of NCDs in LMICs. Reconsidering
the role that is given to stakeholders with significant conflicts
of interest is an important opportunity to rebalance the power
distribution in NCD governance and promote a greater role
for civil society and people living with NCDs, who deserve to
be protected by public health policies. add.12118 2. Suzuki M, Webb D, Small R. Competing frames in global health
governance: an analysis of stakeholder influence on the political
declaration on non-communicable diseases. Int J Health Policy Manag. 2022;11(7):1078-1089. doi:10.34172/ijhpm.2020.257 2022;11(7):1078-1089. doi:10.34172/ijhpm.2020.257 3. Moodie R, Stuckler D, Monteiro C, et al. Profits and pandemics:
prevention of harmful effects of tobacco, alcohol, and ultra-processed
food and drink industries. Lancet. 2013;381(9867):670-679. doi:10.1016/
s0140-6736(12)62089-3 4. Marks JH. The Perils of Partnership: Industry Influence, Institutional
Integrity, and Public Health. Oxford: Oxford University Press; 2019. 4. Marks JH. The Perils of Partnership: Industry Influence, Institutional
Integrity, and Public Health. Oxford: Oxford University Press; 2019. 5. Kickbusch I, Gleicher D. Governance for Health in the 21st Century. WHO
Regional Office for Europe; 2012. 5. Kickbusch I, Gleicher D. Governance for Health in the 21st Century. WHO
Regional Office for Europe; 2012. 6. United Nations. Goal 17: Revitalize the global partnership for sustainable
development [Internet]. New York: UN; [No date]. https://www.un.org/
sustainabledevelopment/globalpartnerships/. Accessed May 9, 2020. 7. Ralston R, Hil SE, da Silva Gomes F, Collin J. Towards preventing
and managing conflict of interest in nutrition policy? an analysis of
submissions to a consultation on a draft WHO tool. Int J Health Policy
Manag. 2021;10(5):255-265. doi:10.34172/ijhpm.2020.52 8. Rinaldi C, van Schalkwyk MC, Egan M, Petticrew M. A framing analysis
of consultation submissions on the WHO global strategy to reduce the
harmful use of alcohol: values and interests. Int J Health Policy Manag. 2021. doi:10.34172/ijhpm.2021.68 8. Rinaldi C, van Schalkwyk MC, Egan M, Petticrew M. A framing analysis
of consultation submissions on the WHO global strategy to reduce the
harmful use of alcohol: values and interests. Int J Health Policy Manag. 2021. doi:10.34172/ijhpm.2021.68 9. Mialon M. An overview of the commercial determinants of health. Global
Health. 2020;16(1):74. doi:10.1186/s12992-020-00607-x 9. Mialon M. An overview of the commercial determinants of health. Global
Health. 2020;16(1):74. doi:10.1186/s12992-020-00607-x 10. Parker LA, Zaragoza GA, Hernández-Aguado I. Promoting population
health with public-private partnerships: where’s the evidence? BMC
Public Health. 2019;19(1):1438. doi:10.1186/s12889-019-7765-2 10. Parker LA, Zaragoza GA, Hernández-Aguado I. Promoting population
health with public-private partnerships: where’s the evidence? BMC
Public Health. 2019;19(1):1438. doi:10.1186/s12889-019-7765-2 References 1. West R, Marteau T. Commentary on Casswell (2013): the commercial
determinants of health. Addiction. 2013;108(4):686-687. doi:10.1111/
|
https://openalex.org/W2186295708
|
https://zenodo.org/record/1434690/files/2115oraj02.pdf
|
English
| null |
A Single Stage Single Constraints Linear Fractional Programming Problem: An Approach
|
Zenodo (CERN European Organization for Nuclear Research)
| 2,015
|
cc-by
| 1,602
|
ABSTRACT In the present paper we present a new method for solving a class of single stage single constraints linear
fractional programming (LFP) problem. The proposed method is based on transformation the objective
value and the constraints also. After reducing the fractional program in to equivalent linear program with
the help of transformation technique, after that we apply Simplex method to find objective value. Numerical
examples are constructed to show the applicability of the above technique Operations Research and Applications : An International Journal (ORAJ), Vol.2, No.1, February 201 Operations Research and Applications : An International Journal (ORAJ), Vol.2, No.1, February 201 A single stage single constraints linear fractional
programming problem: An approach Sapan Kumar Dasa, T. Mandala aDepartment of Mathematics, National Institute of Technology,
Jamshedpur-831 014, Jharkhand, India KEYWORDS Linear fractional programming problem(LFPP); Linear program; Simplex method 1. INTRODUCTION Linear fractional programming problem (LFPP) deals with problems in which objective function
is a ratio of two linear functions. Maximizing the efficiency of an economicsystem leads to
optimization problems whose objective function is a ratio. Linear fractional problems may be
found in different fields such as data development analysis, taxprogramming, risk and portfolio
theory, logistic and location theory [7, 6, 5, 4, 3]. Also,linear fractional programming is used to
achieve the highest ratio of outcome to cost, theratio representing the highest efficiency. Charnes and Cooper [8] proposed several methods for solving linear fractional program by
transforming it to an equivalent linear program. Bitran and Novaes [18] considered updated
objective functions method to solve linear fractional program by solving a sequence of linear
programs whereas Dinkelbach [9] used parametric approach to solve a linear fractional
programming problems. Later on, several authors extended the approach by Dinkelbach [9] to
solve fractional programming problems, e.g., generalized fractional programming problems [10,
11] and the minimum spanning tree with sum of ratios problems [12]. Almogy and Levin [13]
extended the parametric approach of Dinkelbach [9] to solve sum of ratios problems. Falk and
Palocsay [16] showed that the approach in [13] does not always lead to appropriate solutions and
they extended the parametric approach of Dinkelbach to solve sum of ratios, product of ratios and
product of linear functions in [14]. Tammeret al. [15] considered Dinkelbach approach to solve
multiobjective linear fractional programming problems by estimating the parameters. However,
their approach does not necessarily guarantee an efficient solution. Gomes et al. [17] focused on 9 9 multiobjective linear programming problem having weights established some optimality
conditions. multiobjective linear programming problem having weights established some optimality
conditions. During recent years, complexity of problems arising in different fields prompted researchers to
develop efficient algorithms to solve linear fractional programs. Valipouret al. [19] suggested an
iterative parametric approach for solving multiobjective linear fractional programming (MOLFP)
problems. Cambiniet al. [20] reviewed methods for solving biobjective linear fractional
programming. Recently, Tantawy [21] suggested an iterative method based on conjugate gradient
projection method for solving linear fractional programming problem. In the present paper, we first convert linear fractional programming problem (LFPP) to linear
program one with the help of transformation technique. We also present an example to clarify the
proposed method. 1. INTRODUCTION In section 2 some notations and definitions is given while in section 3 we give
the main result with suitable numerical example and a concluding remark is given in section 4. 2. DEFINITIONS AND METHODOLOGY A linear fractional programming problem occurs when a function is minimize or maximize and
the objective function is ratio (numerator and denominator) and the constraints are linear type
function. Consider a linear fractional programming problem: This problem can be applied to find an optimal productivity solution, minimizing or maximizing
the ratio between storage cost and production cost under the storage constraints, Z=the total ratio
cost between storage cost and production cost, 3. Example Consider the following LFP problem First we solve the LFP problem by using our proposed technique to its correspondinglinear
programming problem, First we solve the LFP problem by using our proposed technique to its correspondinglinear
programming problem, Now this problem is our linear programming problem and solved by regular simplex method. We
find the variable is
, 0 and the optimal solution for the relaxed linear programming with optimal
value F(x) = 9/2.This result is same as the result of [22]. The method is very useful because of his
calculations involved are very simple and take least time as compare as other method for solving
linear fractional programming problem. We also solved this problem by LINGO software and
find objective solutions is same as our proposed method result. 2.1 Methodology Transformation of Objective: Transformation of Objective: Multiplying
on both numerator and denominator we have, Multiplying
on both numerator and denominator we have, 10 Operations Research and Applications : An International Journal (ORAJ), Vol.2, No.1, February 2015 Operations Research and Applications : An International Journal (ORAJ), Vol.2, No.1, February 2015 Transformation of constraints: Transformation of constraints: Transformation of constraints: Now consider the linear programming problem from the above transformation of objecttive and
transformation of constraints we have, Now consider the linear programming problem from the above transformation of objecttive and
transformation of constraints we have, 11 11 Operations Research and Applications : An International Journal (ORAJ), Vol.2, No.1, February 2015 Operations Research and Applications : An International Journal (ORAJ), Vol.2, No.1, February 201 Operations Research and Applications : An International Journal (ORAJ), Vol.2, No.1, February 2015 REFERENCES [1]
R. P. Agarwal, I. Ahmad, S. K. Gupta, N. Kailey, Generalized second-order mixed symmetric duality
in nondifferentiable mathematical programming, Abst. Appl. Anal. 2011 (2011) Article ID 103597.4
[2] R. P. Agarwal, I. Ahmad, S. K. Gupta, A note on higher-order nondifferentiable symmetric duality in [2] R. P. Agarwal, I. Ahmad, S. K. Gupta, A note on higher-order nondifferentiable symmetri
multiobjective programming, Appl. Math. Lett.24 (2011) 13081311. [3] K. M. Mjelde, Allocation of resources according to a fractional objective, European J. Oper. Res., 2
(1978) 116-124. [4] A. Charnes, W. W. Cooper, A. Y. Lewin, Data envelopment analysis: Theory, Methodology and
Applications, Seiford L. M. (Ed.), Kluwer Academic Publishers, Boston MA, 1995. [5] A. Charnes, W. W. Cooper, E, Rhodes, Measuring the efficiency of decision making units, European
J. Oper. Res.,2 (1978) 429-444. [6] A. Barros, Discrete and fractional programming technique for location models, Kluwer Academics
Publishers, 1998. [7] E. B. Bajalinov, Linear fractional programming:Theory, Methods, Applications, and Software,
Kluwer Academic Publishers, Boston MA , 2004., [8] A. Charnes, W. W. Cooper, Programming with linear fractional functional, Naval Research Logistics
Quarterly, 9 (1962) 181-186. [9] W. Dinkelbach, On nonlinear fractional programming, Manage. Sci. 13 (1967) 492-498. [10] J. P. Crouzeix, J. A. Ferland, S. Schaible, An algorithm for generalized linear fractional
programming, J. Global Optim. Theory Appl. 47 (1985) 35-49. [11] S. Schaible, J. Shi, Recent developments in fractional programming :single ratio and max-min case,
in: W. Takahashi, T. Tanaka (eds), Proceding of the 3rd international conference in nonlinear
analysis, Yokohama Publisher, Yokohama,(2004) 493-506. y
,
,
,(
)
[12] C. C. Skiscimi, S. W. Palocsay, Minimum spanning trees with sum of ratios, J. lobalOptim. 19
(2001) 103-120. Y. Almogy, O. Levin, A class of fractional programming problems, Oper. Res. 19(1971) 57-67. [14] J. E. Falk, S. W. Palocasy, Optimizing the sum of linear fractional functions, in: C. A. Floudas, P. M. Pardalos (eds), Recent Advances in Global Optimization, Kluwer Academic Publishers, Dordrecht,
(1992) 221-258. [15] K. Tammer, C. Tammer, E. Ohlenderf, Multicriterial fractional optimization, in: J. Guddat, H. T. Jongen, F. Nozicka, G. Nozicka, F. Still, Twilt (eds), Parametric optimization and related topics Iv,
Peter Lang, Berlin, (1997) 359-370. g
[16] J. E. Falk, S. W. Palocsay, Image space analysis of generalized fractional rograms,J. Global Optim., 4
(1994) 63-88. [17] R. O. Gomez, A.R. Lizana, P. R. Canales, Multiobjective fractional programming with generalized
convexity, Top 8 (2000) 97-110. 4. CONCLUSIONS In this paper, we present a transformation method for solving linear fractional programming
problem when the objective function is ratio function and the set of constraints isin the form of
linear inequality. Our proposed method based upon transformation technique. Our new method 12 Operations Research and Applications : An International Journal (ORAJ), Vol.2, No.1, February 2015 Operations Research and Applications : An International Journal (ORAJ), Vol.2, No.1, February 201 can be applied to any linear fractional programming problem,since it is a special thing of the
mathematical program. can be applied to any linear fractional programming problem,since it is a special thing of the
mathematical program. can be applied to any linear fractional programming problem,since it is a special thing of the
mathematical program. [23] V. N. Mishra, Some problems on approximations of functions in Banach spaces,Ph.D. Thesis (2007),
Indian Institute of Technology, Roorkee. [24] V. N. Mishra et al., Inverse result in simultaneous approximation by BaskakovDurrmeyer-Stancu
operations, Journal of Inequalities and Applications (2013) 586. Operations Research and Applications : An International Journal (ORAJ), Vol.2, No.1, February 2015 REFERENCES [18] G. R. Bitran, A. J. Novaes, Linear programming with a fractional objective function, Operation
Research, 21 (1973) 22-29. [19] E. Valipour, M. A. Yaghoobi, M. Mashinchi, An iterative approach to solve mutiobjective linear
fractional programming problems, Applied Mathematical Modelling,38 (2014) 38-49. [20] A. Cambini, L. Martein, I. M. Stancu-Minasian, A survey of bicriteria fractionalproblems, Adv. Model.Optim. 1 (1999) 9-46. . F. Tantawy, A new procedure for solving linear fractional programming problems,Mathematica
nd Computer Modelling 48 (2008) 969-973. [22] S. F. Tantawy, R. H. Sallam, A new method for solving integer linear fractionalprogramming
problems, International Journal of Recent Scienti_c Research 4 (2013)250-253. 13 gy
[24] V. N. Mishra et al., Inverse result in simultaneous approximation by BaskakovDurrmeyer-Stancu
operations, Journal of Inequalities and Applications (2013) 586. 14 14
|
https://openalex.org/W2906780810
|
https://ispan.waw.pl/journals/index.php/sn/article/download/sn.1629/4377
|
Latin
| null |
Subjects of the empress. Wider context of regional history education on East Prussia and identity processes in contemporary Kaliningrad oblast
|
Sprawy Narodowościowe. Seria Nowa
| 2,018
|
cc-by
| 9,228
|
Miłosz Janusz Zieliński Miłosz Janusz Zieliński This is an Open Access article distributed under the terms of the Creative Commons Attribution 3.0 PL Licen-
se (creativecommons.org/licenses/by/3.0/pl/), which permits redistribution, commercial and non-commercial,
provided that the article is properly cited. © The Author(s) 2018.
Publisher: Institute of Slavic Studies, Polish Academy of Sciences CITATION: Zieliński, M. J. (2018).
Subjects of the Empress. Wider Context
of Regional History Education on East Prussia
and Identity Processes in Contemporary
Kaliningrad Oblast.
Sprawy Narodowościowe. Seria nowa, 2018(50).
https://doi.org/10.11649/sn.1629 A b s t r a c t Political, economic and social changes brought about
by the dissolution of the Soviet Union have had a significant
impact on Kaliningrad Oblast, the empire’s westernmost
territory of geostrategic importance. Formerly belonging to
the German province of East Prussia, the region was meant
to become Sovietised completely. The end of the Cold War
led to a complete bankruptcy of such policies. At the turn
of the 80s and 90s the emergence of a grass-roots interest
in the officially forbidden parts of the Oblast’s history made
the question of the relation between its pre-war and post-
war past up-to-date. Its topicality was strengthened after
the 2005-06 commemorations of founding the city of Königs-
berg and Kaliningrad Oblast, in which both central and regional
authorities were heavily involved. This paper aims at identify-
ing how elements of the history of East Prussia have been se-
lected, interpreted and incorporated into the regional history
education course books in contemporary Kaliningrad Oblast. It
argues that the growing criticism of the course books’ con-
tents has been related to Russia’s domestic situation since
the 2012 presidential election and the tensions with the West
after 2014. Both events have hastened the process of forming
and cementing the so-called new Russian conservatism which
has had a growing influence on the Baltic semiexclave. SPRAWY NARODOWOŚCIOWE Seria nowa / NATIONALITIES AFFAIRS New series, 50/2018
DOI: 10.11649/sn.1629
Article No. 1629 This work was supported
by the National Science Centre
(grant number UMO-2012/06/A/HS3/00266).
No competing interests have been declared. MIŁOSZ JANUSZ ZIELIŃSKI
SWPS Uniwersytet Humanistycznospołeczny,
Warszawa
E-mail: milosz@miloszzielinski.pl MIŁOSZ JANUSZ ZIELIŃSKI MIŁOSZ JANUSZ ZIELIŃSKI MIŁOSZ JANUSZ ZIELIŃSKI
SWPS Uniwersytet Humanistycznospołeczny,
Warszawa
E-mail: milosz@miloszzielinski.pl SWPS Uniwersytet Humanistycznospołeczny,
Warszawa E-mail: milosz@miloszzielinski.pl K e y w o r d s: Kaliningrad Oblast; Russian Federation; Soviet
Union; East Prussia; history education; regional identity; na-
tional identity This work was supported
by the National Science Centre
(grant number UMO-2012/06/A/HS3/00266). No competing interests have been declared. Page 1 of 17 S t r e s z c z e n i e Zmiany polityczne, gospodarcze i społeczne spowodowane rozpadem Związku Sowieckiego wywar-
ły głęboki wpływ na obwód kaliningradzki, najdalej wysunięty na zachód obszar imperium o dużym
znaczeniu geostrategicznym. Obwód, wcześniej wchodzący w skład niemieckich Prus Wschodnich,
miał zostać całkowicie zsowietyzowany. Koniec zimnej wojny doprowadził do bankructwa takiej po-
lityki. Na przełomie lat osiemdziesiątych i dziewięćdziesiątych pojawienie się oddolnego zaintereso-
wania oficjalnie zakazanymi częściami historii obwodu spowodowało, że pytanie o stosunek między
jego przeszłością przedwojenną i powojenną stało się bardzo aktualne. Ten stan został wzmocniony
po uroczystościach związanych z założeniem miasta Królewiec i obwodu kaliningradzkiego w latach
2005-06, w które zaangażowały się zarówno władze centralne, jak i regionalne. Niniejszy artykuł ma
na celu prześledzenie, w jaki sposób elementy historii Prus Wschodnich zostały wybrane, zinterpre-
towane i włączone do podręczników poświęconych historii regionu we współczesnym obwodzie ka-
liningradzkim. Autor argumentuje, że rosnąca krytyka treści podręczników związana jest z sytuacją
wewnętrzną Rosji po wyborach prezydenckich w 2012 roku oraz rosnącymi napięciami w stosun-
kach z państwami zachodnimi. Obydwa wydarzenia przyspieszyły proces kształtowania i krzepnięcia
tak zwanego nowego rosyjskiego projektu konserwatywnego, który wywiera coraz większy wpływ
na bałtycką półeksklawę. S ł o w a k l u c z o w e: obwód kaliningradzki; Federacja Rosyjska; Związek Sowiecki; Prusy Wschod-
nie; nauczanie historii; tożsamość regionalna; tożsamość narodowa S ł o w a k l u c z o w e: obwód kaliningradzki; Federacja Rosyjska; Związek Sowiecki; Prusy Wschod-
nie; nauczanie historii; tożsamość regionalna; tożsamość narodowa Introduction C
hanges brought about by the dissolution of the Soviet Union have had a significant
impact on Kaliningrad Oblast, the empire’s westernmost part incorporated in 1945. Created as a rudimentary negation of its pre-war predecessor, Germany’s East
Prussia, the region was meant to become sovietised in every possible aspect. In particu-
lar, its seven-century-long history was to be rewritten from the perspective of Marxism-
Leninism and Russo-Slavic imperialism, discrediting East Prussia as a militarist, aggressive
and anti-Slavic political entity, the destruction of which was an act of historical justice. This
narrative was intended to become deeply rooted into the mindset of the new inhabitants
of the Oblast who came from various parts of the Soviet Union. They were very often
too young to remember the pre-October Revolution period, which additionally intensified
the impact of the new approach (Костяшов, 1996, p. 87; Сологубов, 2012, p. 41). C The end of the Cold War led to a complete bankruptcy of such policies. The emer-
gence of a spontaneous, grass-roots interest in alternative visions of the Oblast’s history,
initially in very limited circles of regional intelligentsia and nomenclature, made the ques-
tion of relations between the pre-war and post-war past of the region increasingly impor-
tant (Дементьев, 2014). The real boom took place when information technologies started
to spread in the city of Kaliningrad and, to some extent, in other parts of the Oblast. It
resulted in a growing number of ordinary citizens taking part in the discussion on what
East Prussia means on the one hand, and what it should mean on the other hand for con-
temporary Kaliningraders. Together with academicians, politicians and NGO activists they Page 2 of 17 Page 2 of 17 started asking questions to what extent the ‘East Prussianness’ could be compatible with
the ‘Sovietness’ and the ‘Russianness’ on regional, local and individual levels. It is now visible that this discussion has become a part of a broader, Russian-wide
discourse on the reinterpretation of both the Russian and the Soviet history, also in terms
of shaping the national and regional history education. In this article I take a close look at
how the 1871-1945 period of the history of East Prussia (from the unification of Germany
until the defeat of the Third Reich in World War Two) has been portrayed in the regional
history course books in Kaliningrad Oblast. Introduction Their contents have been put into both the re-
gional context and the developments in Russia since 2012: the emergence of the so-
called new conservatism and the sharp deterioration of Russia’s relations with the West
after the annexation of Crimea and breaking out of the war in Eastern Ukraine. I have
sought to explain how criticism of the course books was influenced by these changes af-
fecting Russia in general and Kaliningrad Oblast in particular. In order to achieve the above-mentioned goals I have used a variety of research ma-
terial. The background knowledge on the history of East Prussia and Kaliningrad Ob-
last was provided by books by C. Clark (Clark, 2009), Ю. В. Костяшов (Костяшов, 2009),
A. Kossert, R. Traba (Traba, 2005) and A. Sakson (Sakson, 2011). Works by O. Sezneva
(Sezneva, 2007, 2010, 2013) and И. О. Дементьев (Дементьев, 2014) gave a specific look
at how attitudes towards the pre-war past evolved in Kaliningrad Oblast. The central part
of the article revolves around the regional history course books written by Г. В. Кретинин,
А. П. Клемешев, Ю. В. Костяшов and Г. М. Федоров for grades from 6 to 11 (Клемешев, Ко-
стяшов, & Федоров, 2007; Кретинин, 2007a, 2007b). In order to confront the contents of the course books with the overall context of in-
ternal developments in Russia since the turn of 2011 and 2012, I have used a number
of articles from the presidential campaign published by V. Putin in various Russian media
outlets (Путин 2012a; Путин 2012b; Путин 2012c) and studies analysing them, such as
those on the ‘new conservatism’ originating from the Warsaw-based Centre for Eastern
Studies (Rodkiewicz & Rogoża, 2015; Rogoża, 2013). Criticism of their contents has pre-
dominantly been present on regional web portals, but also in the Russian-wide press,
such as “Литературная газета”. How East Prussia (re)appeared in Kaliningrad Oblast When World War Two came to an end East Prussia was in ruins. As the very first prov-
ince of the pre-1937 Germany that the Red Army entered, it suffered not only from regu-
lar military actions, but also from the aftermath events. The vendetta of the Soviet sol-
diers was stoke up by the authorities. In the wartime and the post-war Soviet propaganda
East Prussia was depicted as, for instance, ‘the lair of the fascist beast’ (Костяшов, 2003,
p. 11) and ‘German Empire’s watchdog in the East’ (Костяшов, 2003, p. 14). Such an ag-
gressive way of portraying the province had calamitous effects on civilians who did not
escape before the Red Army as well as on the material culture. Approximately 100,000 Germans—less than 10 per cent of the pre-war population, re-
mained in the province until 1950. They were resettled to the Soviet occupational zone
of Germany (Костяшов, 1996, p. 83). New inhabitants came mostly from the Russian, Be-
larusian and Ukrainian Soviet Republics. They were predominantly young and did not re- Page 3 of 17 Page 3 of 17 member the times prior to 1917. Speaking in terms of civilizational experience their new
home was completely different from what they were used to. For authorities in Moscow
the Oblast was thus a perfect place for creating Homo Sovieticus (Heller, 1988). Interest in discovering the pre-Soviet roots of the region appeared already in the second
generation of the Kaliningrad inhabitants and grew alongside the scope of liberty in the coun-
try (Дементьев, 2014, pp. 183–184). During perestroika it took the form of hobbyists-diggers
who semi-legally excavated old pieces of cutlery, crockery or bottles (Sezneva, 2007). Nev-
ertheless, a deeper interest in the past was shared only by a limited circle of regional intel-
ligentsia and a few members of the nomenclature (Дементьев, 2014, pp. 188–192). The situation changed following the dissolution of the Soviet Union (Хоппе, 2005,
p. 199). After the initial period of economic downturn and serious social challenges
the notion of the pre-war history started to find its place in the everyday life of Kalinin-
graders. It even gained a nationwide attention in 2005-2006 alongside the official celebra-
tions of 700th Anniversary of Königsberg and 70th Anniversary of Kaliningrad. How East Prussia (re)appeared in Kaliningrad Oblast President
Vladimir Putin met with Chancellor Gerhard Schröder and President Jacques Chirac in
Kaliningrad, where he emphasized the European roots of the city and supported the idea
of rebuilding its historic centre (Siegień, 2016, p. 116). The 2005-06 events marked the moment when the notion of East Prussia evolved
from a hobby of the few to an inseparable part of a broader public debate on how it
should be rediscovered and interpreted on a regional level. The once forbidden ‘Prus-
sian spirit’ quickly became a full-featured element of the main quotidian topic on various
levels. Gradually a growing number of institutions, starting from the governor’s office and
other regional governmental bodies, became involved in shaping the ‘East Prussian part’
of the political, economic and social agenda in contemporary Kaliningrad Oblast. The captivation of the pre-war history topic was further strengthened by a rapid devel-
opment of information technologies. Regional portals, discussion fora and social networks
became platforms for intensive dialogue between individuals. Favourable climate for speak-
ing about pre-1945 times openly and publicly became a trigger for the idea of introducing
a regional history course in the secondary and high school education system in the Oblast. 1
The University was founded in 1967 under the name of Kaliningrad State University. In 2005 it was renamed
to Immanuel Kant Russian State University by President Putin and Chancellor Schröder. In 2010 President
Medvedev renamed it to Immanuel Kant Baltic Federal University.
2
In this article they will be jointly referred to as ‘authors’ 2
In this article they will be jointly referred to as ‘authors’. to
a ue
a t
uss a
State U
e s ty by
es de t
ut
a d C a ce o Sc
öde
0 0
es
Medvedev renamed it to Immanuel Kant Baltic Federal University.
2 to Immanuel Kant Russian State University by President Putin and Chancellor Schröder. In 2010 Presiden
Medvedev renamed it to Immanuel Kant Baltic Federal University.
2 1
The University was founded in 1967 under the name of Kaliningrad State University. In 2005 it was renamed
to Immanuel Kant Russian State University by President Putin and Chancellor Schröder. In 2010 Presiden Zapadnyi krai: the regional history course books and initial
criticism The course on the history of the Western Land (Западный край) or Western Rus-
sia (Западная Россия) was created by a group of academicians from the Immanuel Kant
Russian State University:1 Кретинин, Г. В. (classes 6-9), А. П. Клемешев, Ю. В. Костяшов,
Г. М. Федоров (classes 10-11).2 The outline of the course was presented in the document
entitled the Programme of Public Education on Regional History (Программа общего
образования по истории края; Кретинин & Строганова 2006). It served as a basis of three
course books: from prehistoric times to the 18th century; until the beginning of World
War Two; since 1939-1941 until 2006. The last part tells the story of East Prussia being
conquered by the Red Army in 1944-45, Soviet Kaliningrad Oblast being emerged and Page 4 of 17 Page 4 of 17 the way developments in the region shaped after the dissolution of the Soviet Union. The
testing period of the course was launched in September 2006 in 36 schools attended
by 2,250 students (Корвин, 2007). By large, while discussing the Russo-Prussian relations in the 19th century, authors
highlight their positive aspects, such as Prussia’s neutrality during the Crimean War,
which helped Russia limit its isolation in Europe. St. Petersburg returned this favour when
it did not oppose the unification of Germany under the reign of the Hohenzollern (Крети-
нин, 2007b, pp. 139–142). In the overall complexity of Russo-German relations East Prus-
sia played the role of a marketplace and was an area of relatively high level of freedom
where various political ideas were free to emerge. Authors give an example of socialist
activists whose actions contributed to preparing the ground for the 1905 revolution in
Russia. Although they were persecuted by German law enforcement bodies, they could
count on a fair trial and successfully defend their views (Кретинин, 2007b, pp. 139–140). Thus, intelligentsia felt comfortable in the city of Königsberg where atmosphere of free
thought prevailed over the militaristic Prussian government (Кретинин, 2007b, p. 157). Much complicated reasons for the outbreak of war in 1914 were explained very brief-
ly. Authors took laconic notes of deterioration of the Russo-German relations in 1891-
1894 (Кретинин, 2007b, pp. 141–142). When the conflict finally started in 1914 military
actions in East Prussia were interrelated with those on the Western Front. The Russian
army entered the province in order to fulfil its duties towards France (Кретинин, 2007b,
p. 156). Zapadnyi krai: the regional history course books and initial
criticism The main challenges that East Prussia and the government in Berlin had to face per-
tained to stabilise the political and economic situation of the province. Thanks to inten-
sified efforts and considerable financial support, authors write, it was finally achieved
(Кретинин, 2007b, pp. 185–187). It was also possible thanks to economic cooperation
between Germany and Soviet Russia after the Treaty of Rapallo in 1922 when bilateral
diplomatic relations were established. Soviet economic presence in East Prussia was vis-
ible notably through the East German Trade Fair (the Ostmesse; Восточная ярмарка) in
Königsberg (Кретинин, 2007b, pp. 188–190). The end of the 1930s, however, was marked with increased tensions in Europe which
resulted in another world war. Authors of the course books start the wartime story
of East Prussia from 1941, i.e. from the beginning of the so-called Great Patriotic War. They also mention, however, that the province took part in the war campaign against
Poland in September 1939 (Клемешев et al., 2007, pp. 3–4). Although East Prussia is de-
scribed as place d’armes (плацдарм) of Germany’s aggression against the Soviet Union,
the war seemed to be distant for the inhabitants of the province for three years, with
the exception of rare air bombings of the Soviet aviation (Кретинин, 2007b, p. 200). The third part of the course books starts from describing military actions in East
Prussia in 1944-1945 (with a short introduction referring to September 1939), devoting
much space to Soviet war heroes: General Ivan D. Chernyakhovsky, Marshall Konstan-
tin K. Rokossovsky, as well as lower rank heroes Alexey I. Tazayev and Sergey I. Gu-
sev, who were killed in action in the province. Particular attention is given to the siege
of Königsberg, led by General Alexandr M. Vasilevski (Клемешев et al., 2007, pp. 13–18). The end of the war was followed by the introduction of the Soviet military administra-
tion in East Prussia. Authors show a considerable deal of criticism towards its actions. They touch upon the controversial topic of the so-called bounty hunters (трофейные ко-
манды) who did great harm to the regional economy, impeding its reconstruction. As it
is explained in the course books, such actions were tolerated due to uncertain future
of the territories conquered by the Red Army. Zapadnyi krai: the regional history course books and initial
criticism The quick assault on East Prussia in summer of 1914 is described as a strategic
mistake resulting in Russia’s defeat in the First Battle of the Masuria Lakes (Кретинин,
2007b, pp. 165–166). Authors even write that “the credit has to be given to the German
commandment” (надо отдать должное германскому командованию) for the way that lib-
erating East Prussia was carried out (Кретинин, 2007b, p. 172). At the same time authors
note the importance of the 1914 campaign in the German nationalist symbolic of the in-
terwar period. They give the Tannenberg Memorial as an example (Tannenberg Denkmal;
Кретинин, 2007b, p. 166). The course books touch upon the behaviour of Russian forces in East Prussia. Authors
acknowledge that there might have been incidental atrocities committed towards civil-
ians, yet they underline that the majority of Russian troops behaved orderly. To strength-
en this statement authors present testimonies of German citizens who not only confirm
this correct stance but also oppose other points of view overemphasizing the alleged
bestiality of Russians on East Prussian soil (Кретинин, 2007b, pp. 166–169). Authors em-
phasize that the question of relations between armed forces and civilians during wartime
objectively belongs to the list of the most difficult ones and that “East Prussia was no
exception” (Кретинин, 2007b, p. 169). Authors seek to highlight the universal atrocity affecting the mankind in general, re-
gardless of ethnic or political differences. Thousands of casualties of the first mass-scale
conflict in history of both German and Russian nationalities were buried on numerous
sites in East Prussia. The course books show the example of a Russo-German graveyard
in the village of Lasdehnen (contemporary Kranoznamensk) which has been preserved
until the present day (Кретинин, 2007b, p. 163). In Kaliningrad Oblast there are also sites
where exclusively Russian soldiers were buried (Кретинин, 2007b, p. 173). Consequences of the war are described as being of both real and symbolic nature. The
latter is related to the formal end of the Kingdom of Prussia (which in the course books is
referred to as the Duchy of Prussia—Герцогство Пруссия). Authors write that East Prussia as Page 5 of 17 a separate entity was emblematic of “Germany’s claims towards the Baltic states, Poland,
[as well as] Russian, Belarusian and Ukrainian lands” (Кретинин, 2007b, p. 179). It did not,
however, cancel the claims themselves even when the Weimar Republic was established. The post-2012 context The post-2012 context The internal situation of the post-Yeltsin Russia has been increasingly determined by poli-
cies formed on the central level. This phenomenon has reversed the inertia of decentrali-
sation and partial regionalisation observed since the last years of existence of the Soviet
Union (Marciniak, 2001). Back then some regions gained considerable factual autonomy
due to an insignificant control exercised by Moscow. When Vladimir Putin came to power
the role of the vast majority of regional authorities quickly changed from moderately in-
dependent players to plain executors of the Kremlin-shaped policies. It has had serious
consequences on all subjects of the Russian Federation and cannot be explained briefly
without making major simplifications. During the 2012 presidential campaign Vladimir Putin published seven articles on dif-
ferent aspects of Russia’s present and future, out of which at least three tackled history
and culture related issues (Путин, 2012a, 2012b, 2012c). Putin sketched the desired path
of Russia’s development and the set of values that it should abide by. He underlined
the positive role of the strong state apparatus which had been deeply rooted in the Rus-
sian political culture. He also stressed the need for gradual changes and opposed radical
moves (Rodkiewicz & Rogoża, 2015, pp. 8–9). Later on, Putin conceptualised and refined
his points, paving the way for the so-called new conservative project. He suggested that
Russia is a distinctive civilizational centre and should pursue independent political and cul-
tural policies, in particular within the post-Soviet space (Путин, 2012a). Putin’s words were soon forged into concrete actions, notably, to further strength-
en soft power instruments that Russia had started to develop couple of years earlier. This tendency was envisaged in the 2013 Russia’s foreign policy concept (“Concept
of the Foreign Policy”, 2013). In order to strengthen the Russian statehood and its ties
with Russians living abroad, an important role was given to the Russian Orthodox Church,
traditionally closely linked with the authorities, and the Russkiy Mir Foundation estab-
lished in 2007. The second game-changer was the outbreak of the so-called Revolution of Dignity in
Ukraine and the events that followed—annexation of Crimea and war in Donbas. They re-
sulted in sharp deterioration of Russo-Western relations, including sanctions imposed by,
among others, the European Union and the United States. It coincided with the economic
slowdown in Russia caused by structural macroeconomic shortcomings, such as corrup-
tion and mismanagement. Zapadnyi krai: the regional history course books and initial
criticism The course books also encompass the facts about the lives of that part of the Ger-
man population that did not flee their homes, consisting mostly of women, children and
elders. Authors acknowledge harsh conditions inhabitants had to withstand, including
constant problems with provisioning and forced labour, for instance, repairing roads and
dismantling ruins. Although their situation was difficult, Soviet authorities allowed them
to perform religious practices in churches which was forbidden for Orthodox newcomers
(Клемешев et al., 2007, pp. 24–25). In general, the course books tell the story of 1944-1945 and the first post-war years
in a very objective and unbiased way. While accepting the geopolitical consequences
of the war they do not omit negative aspects of the new political, economic and social
circumstances. Students reading the book are likely to understand that history is never
either just black or white, no matter how trivial such a remark would sound. Authors em-
bark from regional rather than national history narrative. They accept the fact that until
1945 East Prussia’s administrative language was German. One might wonder that is why
they consequently use German toponyms (written in Cyrillic) throughout the pre-1945
chapters and do not mention their contemporary Russian equivalents. Page 6 of 17 The post-2012 context Together with the growing atmosphere of the ‘new Cold War’,
the Kremlin turned to well-known tools in order to gain popular support, such as the myth
of a besieged fortress (осаждённая крепость). According to it, for centuries Russia has
been surrounded by enemies who have tried to weaken and humiliate the country. Citi-
zens of contemporary Russia should once again unite around their leader to repel the ag-
gressors (Колесников, 2015). Confrontation invoked by the events of 2012-2014 became visible in Kaliningrad Oblast,
although with some delay in many important areas of socio-political life of the semiex-
clave. At first, economic downturn was stronger in the region than in Russia in average. This created a theoretical danger that Kaliningraders might be willing to show their discon-
tent by taking to the streets similar to 2009 and 2010, which finally did not take place. For
a number of reasons they either supported Kremlin’s policies or remained passive. In Moscow’s eyes geostrategic importance of the Oblast prevailed over its coopera-
tion potential towards its immediate neighbourhood. Its military potential was signifi- Page 7 of 17 cantly strengthened (Wiśniewska, Domańska, Strzelecki, Żochowski, Wilk, & Menkiszak,
2016). Altogether, the region once again was placed at the frontline of political or any
other kind of confrontation with the West. Managing the crisis in the Oblast became of significant importance for central authori-
ties in view of the 2018 FIFA World Championship, which will be co-hosted by Kalinin-
grad. Ensuring smooth preparations for the event was the main reason for major shifts in
the regional administration. Governor Nikolay Tsukanov was temporarily replaced by a si-
lovik Georgi Zinichev. Later on, Zinichev yielded for Anton Alikhanov, a young technocrat
whose main task was to stabilise economic situation of the region and ensure swift im-
plementation of Kremlin’s policies. His first months in office confirmed this reasoning. Both new governors limited regional authorities’ engagement in issues connected
with the pre-war legacy of the Oblast, such as the reconstruction of the historical cen-
tre of Kaliningrad and preservation of pre-1945 buildings. In fact, they narrowed down
the official historical agenda to the elements directly corresponding to the ‘new Russian
conservatism’. Alikhanov took an active part in numerous events commemorating Rus-
sia’s presence in East Prussia; for instance, during the Seven Years’ War of 1756-1763,
when Russian troops came to the province both as an enemy and as an ally. The post-2012 context In August
2017, Alikhanov unveiled a monument devoted to General Field Marshall Pyotr Rumyant-
sev (Сумарокова, 2017). Two weeks later he opened a renewed monument of the bat-
tle of GrossJägersdorf (Гросс-Егерсдорф; contemporary village of Междуречье). During
the ceremony he said: “Here a whole host of Russian war commanders has written their
names into the history of this soil with golden letters”3 (“Антон Алиханов открыл”, 2017). 3
Здесь целая плеяда российских военных начальников вписала золотыми буквами свои имена в историю этой
земли.
4
Внедрение „Истории Западной России” в качестве регионального компонента в нынешнем объёме (34 часа
в год) неизбежно будет способствовать воспитанию у учащихся сепаратизма и прогерманской ориентации Criticism of the course books The course books were met with significant criticism the instant they were introduced. Main objections pertained to the allegedly pro-German perspective from which the au-
thors told the history of East Prussia. In the books it was acknowledged that throughout
seven centuries of its existence East Prussia was closely linked with the German culture,
language and politico-legal reality. Thus, critics argued: “The introduction of the History
of Western Russia as a regional component in the current time frame (34 hours a year)
will inevitably provide an incentive to promote separatism and a pro-German orientation
among the students”4 (Корвин, 2007). It was, however, not until 2014-2015 that the criticism gained on strength. Tatyana
Shabayeva expressed her reservations towards the course books in “Литературная газе-
та” in early 2015. While avoiding politically motivated arguments, Shabayeva wrote: Please read [in the course book for grades 8-9]: “The beginning of the war [East Prussian
people] welcomed. The war quickly moved eastward from the borders of the Reich. There
seemed to be no threat to the province. The bombings of the Soviet aviation were a sad
reminder of the war.” Who should be compared to a young man who learned the history
of the same people who “welcomed the beginning of the war with satisfaction”? Will he be 3
Здесь целая плеяда российских военных начальников вписала золотыми буквами свои имена в историю этой
земли. 4
Внедрение „Истории Западной России” в качестве регионального компонента в нынешнем объёме (34 часа
в год) неизбежно будет способствовать воспитанию у учащихся сепаратизма и прогерманской ориентации Внедрение „Истории Западной России” в качестве регионального компонента в нынешнем объёме (34 часа
в год) неизбежно будет способствовать воспитанию у учащихся сепаратизма и прогерманской ориентации Page 8 of 17 Page 8 of 17 able to feel the connection, the continuation between him and the Soviet aviation, which so
unpleasantly destroys the peace of the province? able to feel the connection, the continuation between him and the Soviet aviation, which so
unpleasantly destroys the peace of the province? The authors of “The History of Western Russia” are apparently not worried. The subject goes
to the textbook for grades 10-11: “The war came unexpectedly. On 17 August 1944, the Sovi-
et forces reached the border between the USSR and Germany near Schirwindt [contemporary
village of Kutuzovo].” The children of the Kaliningrad region are again suggested to feel as if
they were on the German side. Criticism of the course books Yet it was for the Germans in 1944 that “the war came unex-
pectedly” (which is a bit strange in the face of “nasty bombardments carried out by the So-
viet aviation”) (Шабаева, 2015).5 At that time some voices spoke about the alleged process of Königsbergization (which
might also be referred to as Germanisation) of Kaliningrad Oblast as a (geo)political rather
than cultural phenomenon (Выползов, 2016). It is significant that they were formulated
shortly after the sharp deterioration of the Russo-Western relations, suggesting that ex-
ploration of the pre-war past had been inspired by external forces intending to weaken
the “Russianness” of the Oblast. These voices devoted little attention to formulating
a positive counter-agenda. The new wave of criticism reached its peak in mid-2017 when the tenth anniversary
of introducing the regional history course gave a pretext for their deeper assessment. In order to present main arguments raised, I have used the round table dedicated to
the course books, which took place in July 2017, as well as some public statements
made by prominent figures in Kaliningrad Oblast. At the round table, Georgiy Biryukov, protoiereus of the Russian Orthodox Church in
Kaliningrad Oblast, described the course and the course books as “categorically harm-
ful”. He referred to the books in terms of civilizational differences between the East Prus-
sian and Soviet/Russian periods of the region’s history, as well as to the books lacking po-
litical and legal explanations why the Soviet Union conquered East Prussia. He underlined
that, in particular, the authors did not include references to: • Churchill’s conversation with Soviet Ambassador Ivan Mayski in 1941, in which the for-
mer proposed to split Germany and separate East Prussia; • Churchill’s conversation with Soviet Ambassador Ivan Mayski in 1941, in which the for-
mer proposed to split Germany and separate East Prussia; • the Tehran and Yalta conferences where the fate of Königsberg was discussed; • the law of the Union Control Council in Germany of 1947 “On the Elimination
of the Prussian State” (“История Западной России способствует”, 2017). Similar arguments were expressed by Vladimir Shulgin of the Russian National Line:6 Similar arguments were expressed by Vladimir Shulgin of the Russian National Line:6 We warned as early as in 2006 that the textbooks “The History of Western Russia” did not
meet the national interests of Russia. Page 9 of 17 5
Вчитайтесь: «Начало войны население встретило с удовлетворением. Боевые действия вскоре
переместились от границ рейха на восток. Казалось, провинции ничто не угрожает. Неприятным
напоминанием о войне были редкие бомбардировки советской авиации». С кем должен соотносить себя
молодой человек, если до сих пор он изучал историю того самого населения, которое «начало войны
встретило с удовлетворением»? Сможет ли он ощутить связь, преемственность с советской авиацией,
которая так неприятно тревожила спокойствие провинции? Авторов «Истории Западной России» это,
кажется, не заботит. Тема перетекает в учебник 10–11-х классов (авторы – А.П. Клемешев, Ю.В. Костяшов,
Г.М. Фёдоров): «Война пришла неожиданно. Советские войска в районе Ширвиндта 17 августа 1944 года
вышли на границу между СССР и Германией». Снова детям Калининградской области предлагается
почувствовать себя на стороне Германии. Это для немцев в 1944 году «война пришла неожиданно» (что
немного странно после «неприятных бомбардировок советской авиации»).
6
Русская народная линия (Russian National Line) is an “Orthodox informational agency” which was founded
in St. Petersburg in 2010 after some members of the “Russian Line” (founded in 2008) left the organisation.
The Russian National Line represents ethno- and religion-nationalist point of view on current developments
in Russia and its close neighbourhood. It is a fierce opponent of the presence of Western civilisation in the
Orthodox world. 5
Вчитайтесь: «Начало войны население встретило с удовлетворением. Боевые действия вскоре
переместились от границ рейха на восток. Казалось, провинции ничто не угрожает. Неприятным
напоминанием о войне были редкие бомбардировки советской авиации». С кем должен соотносить себя
молодой человек, если до сих пор он изучал историю того самого населения, которое «начало войны
встретило с удовлетворением»? Сможет ли он ощутить связь, преемственность с советской авиацией,
которая так неприятно тревожила спокойствие провинции? Авторов «Истории Западной России» это,
кажется, не заботит. Тема перетекает в учебник 10–11-х классов (авторы – А.П. Клемешев, Ю.В. Костяшов,
Г.М. Фёдоров): «Война пришла неожиданно. Советские войска в районе Ширвиндта 17 августа 1944 года
вышли на границу между СССР и Германией». Снова детям Калининградской области предлагается
почувствовать себя на стороне Германии. Это для немцев в 1944 году «война пришла неожиданно» (что
немного странно после «неприятных бомбардировок советской авиации»). 7
Мы ещё в 2006 году предостерегали, что учебники «История западной России» не отвечают национальным
интересам России. Этот курс вместо русской, российской истории рассматривает историю немецкую,
причём на ограниченном круге источников, который не включает себя источники, говорящие о тысячелет-
ней войне Запада с Византийской цивилизацией. Этот курс писали специалисты по зарубежной истории
и один географ. Нет ни одного учёного по отечественной истории. Даже этот факт — показатель ориен-
тации авторов курса. Если этот курс не исправить, он и дальше будет оказывать воздействие не только
на школьников и студентов, но и на тех, кто готовится стать офицерами, защитниками Родины.
8
Regional NGO Russian Community of Kaliningrad Oblast (Региональная общественная организация «Русская об-
щина Калининградской области») was registered only in July 2015 which alone ties the organisation with new
developments on Russian political and social scene after 2014.
9
Преподавание курса «История западной России» в нынешнем виде размывает этнонациональную
идентичность русских людей, обучающихся в калининградских школах, что приводит к ослаблению их
общегражданской российской идентичности.
10 In fact, Klemeshev only co-authored the last part of the series Criticism of the course books This course, instead of Russian [both русская and
российская история] history, tells the history of Germany and it does so based on a limited 5
Вчитайтесь: «Начало войны население встретило с удовлетворением. Боевые действия вскоре
переместились от границ рейха на восток. Казалось, провинции ничто не угрожает. Неприятным
напоминанием о войне были редкие бомбардировки советской авиации». С кем должен соотносить себя
молодой человек, если до сих пор он изучал историю того самого населения, которое «начало войны
встретило с удовлетворением»? Сможет ли он ощутить связь, преемственность с советской авиацией,
которая так неприятно тревожила спокойствие провинции? Авторов «Истории Западной России» это,
кажется, не заботит. Тема перетекает в учебник 10–11-х классов (авторы – А.П. Клемешев, Ю.В. Костяшов,
Г.М. Фёдоров): «Война пришла неожиданно. Советские войска в районе Ширвиндта 17 августа 1944 года
вышли на границу между СССР и Германией». Снова детям Калининградской области предлагается
почувствовать себя на стороне Германии. Это для немцев в 1944 году «война пришла неожиданно» (что
немного странно после «неприятных бомбардировок советской авиации»). Русская народная линия (Russian National Line) is an “Orthodox informational agency” which was founded
in St. Petersburg in 2010 after some members of the “Russian Line” (founded in 2008) left the organisation. The Russian National Line represents ethno- and religion-nationalist point of view on current developments
in Russia and its close neighbourhood. It is a fierce opponent of the presence of Western civilisation in the
Orthodox world. Page 9 of 17 range of sources, which does not include those referring to the millennial war of the West
against the Byzantine Civilisation. This course was written by foreign history specialists and
one geographer. There was not a single Russian history specialist. Even this fact is an indica-
tor of what the point of view of the authors of the course was. If this course is not corrected,
it will continue to have an impact not only on schoolchildren and students, but also on those
who are getting ready to become officers and defenders of the Motherland (“Если не станем
противостоять в Калининграде”, 2017).7 Maxim Makarov, President of the Russian Community of Kaliningrad Oblast Regional
NGO,8 pointed to the relation between the ethno-national and civic identity of Russians. Criticism of the course books According to him, the former should be strengthened in order to stop the process of blur-
ring and weakening the latter, also through the course books which has a negative im-
pact on all of those attending schools in the Oblast9 (“Если не станем противостоять в
Калининграде”, 2017). Among those taking part in the discussion was Andrey Klemeshev, rector of the Fed-
eral Baltic University in Kaliningrad, who is often referred to as the editor of the whole
series of the course books by his critics.10 In 2017 he admitted that the current course
should be replaced by a new one that will reflect the reality better. In particular, it should
take into consideration the post-war (and, one might suspect, post-Cold War) geopolitics. Klemeshev underlined that the whole programme of the regional history was created
very quickly during a difficult period. Nowadays, while the new programme is being de-
veloped, other events and processes should be stressed in the next generation of course
books (“Андрей Клемешев: Учебник”, 2017). One might wonder what is meant by “other events and processes”. An indication
of that has been given by actions taken by regional government officials at least since
2016. They mostly revolve around traces of the Russian and Soviet presence in East Prus-
sia before 1944-45. In particular, they are related to the military actions during the Seven
Years’ War, the Napoleonic Wars, as well as the World War One. For example, during
the first of the above-mentioned conflicts all of East Prussia was conquered by Russian
troops. It was then that Immanuel Kant and other citizens of Königsberg swore allegiance
to Empress Elisabeth. Conclusion For a number of years the regional history course taught in schools of Kaliningrad Ob-
last were met with just a moderate criticism. Arguments of its anti-Russian nature were
raised by a limited circle of institutions and individuals, usually belonging to radical circles. For a number of years the regional history course taught in schools of Kaliningrad Ob-
last were met with just a moderate criticism. Arguments of its anti-Russian nature were
raised by a limited circle of institutions and individuals, usually belonging to radical circles. Page 10 of 17 For numerous reasons it did not lie in the interest of the central and regional Russian
authorities to stop or control the revival of the East Prussian narrative in the Oblast. In
the last two to three years the wave of pessimism has become much stronger. Critics
who reprimanded the manner in which the last years of the existence of East Prussia and
the foundation of Kaliningrad Oblast were portrayed in the regional history course books
have embarked from ideological positions which cannot be understood without placing
them into the Russian-wide context. This can be explained in various ways. After more than a decade-long period of grow-
ing interest towards exploring elements of the East Prussian past and attempts to incor-
porate them into the regional history education, opponents of this process started to voice
their concerns openly and frequently. According to them, some parts of the ‘East Prus-
sian myth’ have come at the expense of the official narrative of the history of the Russian
state. Critics have argued that the emergence of the myth and anchoring it into the eve-
ryday life of ordinary Kaliningraders has been inspired by actors and events from the out-
side of the region. It has been strictly related to the domestic developments of Russia, as
well as to its international situation. The new conservative project pursued by the central Russian authorities has gradu-
ally transposed into numerous parts of the everyday life of ordinary Russians on regional
and local levels. One of the areas of confrontation that the new conservatism suggests is
the alleged set of tools that enemies of Russia have at their disposal. The Kremlin does
not send direct, specific and topic-oriented directions. It rather cares for setting the stage
for lower level actions which would fill it with proper content. Page 11 of 17 Conclusion The books tell a neutral story of a region that was subject to, willingly or not, supraregion-
al events and processes. They do not try to suggest that throughout the majority of time
East Prussia’s existence was harmful to the interests of Russia or the Soviet Union. It
was only the World War Two that led to geopolitical changes, but even then East Prussia
was merely a passive participant in the aggression planned and put into motion by au-
thorities in Berlin. One can argue that the lack of blatant “pro-Russianness” or “anti-Westernness” in
the course books was easier to accept even five years ago. Along with growing assert-
iveness of Moscow’s foreign policy it became less and less sustainable. When tensions
reached their critical point in 2014, the hitherto framework of the Russo-Western coop-
eration took a strong hit. Since then notions such as the Königsbergization have gained
popularity in regional media mostly because they have corresponded to the post-2014
Kremlin-shaped national agenda which has heavily exploited the narrative of a ‘besieged
fortress’ and a civilizational conflict. Under such circumstances there is little place for presenting the history of a region
which objectively is not deeply embedded into the traditionally understood “Russian-
ness”. Such a sharp reaction of those opposing the narrative present in the course books
also points to the fact that there has been a well-established feeling in the region that
the level of cultural, religious and social integration between Kaliningrad Oblast and ‘main-
land’ Russia has to be increased. This feeling is probably shared by the central authorities. From their point of view, such policies serve predominantly geopolitical and geostrategic
purpose. Thus, the criticism against the course books is fully in line with the current pace
and direction of developments in Russia, although, it is perceived somewhat differently
on regional and central levels. Nevertheless, it transposes into similar actions and tools
used on the ground, such as engagement of the Russian Orthodox Church and numerous
cultural organisations into the work on the ground. The course books on the pre-1945 history of Kaliningrad Oblast will probably be re-
written as a part of a larger programme encompassing over 40 of such publications Rus-
sia-wide. The contents of them, however, will not disappear. The reason for this is that
the presence of the pre-war past far exceeds the narrow educational system framework. Conclusion This is why this rhetoric has
been tailored in order to include regional specifics, such as history, social and economic
situation, developmental challenges etc. In the case of Kaliningrad Oblast, its historical
background and proximity to the West (the EU and NATO states) make the core of the re-
gional face of the new conservative agenda. The region is believed to be susceptible to
the foreign influence which forces the authorities and other institutions close to them to
take countermeasures. The discourse on the pre-war history of Kaliningrad Oblast is peculiar in this regard as it
belongs to the dual-use category, depending on the overall climate of the Russo-Western
relations. During Tsukanov’s term in office the question of regional history was constantly
present in the public sphere. It even got envisaged in discussions about concrete projects,
such as the reconstruction of the historical centre of Kaliningrad or renovation of a few re-
maining pieces of pre-war architecture. Mainly because of these actions the Oblast could
be perceived as a region close to Europe which looks at its East Prussian (Western) roots
and takes care of them for numerous reasons. Among them, economic development prob-
ably played the most important role for regional authorities. Although the results of this
discussion have so far been meagre, they have unleashed a considerable social potential in
various groups of the region’s population which became visible in many different spheres. After the 2012-2014 developments in and around Russia the above-mentioned fa-
vourable conditions for exploring the pre-war past of Kaliningrad Oblast came to an end. Changes at the top of the regional administration have depicted that further explora-
tion of the pre-war legacy will not rank high on the new governor’s political agenda. On
the contrary, the time has come to highlight the Russian and Soviet elements of the his-
tory of the Oblast. Not only has it become visible in the new initiatives, but it has also af-
fected the ongoing projects and is likely to affect the existing policies. History education
is undoubtedly one of them. Page 11 of 17 In this context, it needs to be stated that the course books present a highly unbiased
story of how East Prussia existed in pre-1945 Germany. Their authors focus on the prov-
ince itself rather than on German or Russian nation-state historiographical perspectives. Clark, C. (2009). Prusy: Powstanie i upadek: 1600-1947. (J. Szkudliński, Trans.). Warsza-
wa: Wydawnictwo Bellona.
Concept of the Foreign Policy of the Russian Federation. Approved by President
of the Russian Federation V. Putin on 12 February 2013. (2013). Retrieved July 20, Conclusion It will remain one of the main topics in the regional media, as well as in everyday talks
and activities of ordinary Kaliningraders. On the other hand, the discussion in the spirit
of discovering and understanding the pre-war past in a non-biased and non-politically mo-
tivated way will remain being affected by Russia-wide and international circumstances. Where it will bring the debate as a whole is yet to be observed. Bibliography Page 12 of 17 Page 12 of 17 2018, from http://www.mid.ru/en/foreign_policy/official_documents/-/asset_publisher/
CptICkB6BZ29/content/id/122186 Heller, M. (1988). Cogs in the wheel: The formation of Soviet man. New York, NY: Alfred
A. Knopf. Kossert, A. (2009). Prusy Wschodnie: Historia i mit. (B. Ostrowska, Trans.). Warszawa:
Wydawnictwo Naukowe Scholar. Marciniak, W. (2001). Rozgrabione imperium: Upadek Związku Sowieckiego i powstanie
Federacji Rosyjskiej. Kraków: Wydawnictwo Arcana. Rodkiewicz, W., & Rogoża, J. (2015). Potemkin conservatism: An ideological tool
of the Kremlin. Warszawa: Ośrodek Studiów Wschodnich. Rogoża, J. (2013). Adjustment in Putin’s state: Under the banner of conservatism. Retrieved
July
20,
2018,
from
https://www.osw.waw.pl/en/publikacje/analy-
ses/2013-12-18/adjustment-putins-state-under-banner-conservatism Sakson, A. (2011). Od Kłajpedy do Olsztyna: Współcześni mieszkańcy byłych Prus
Wschodnich: Kraj Kłajpedzki, Obwód Kaliningradzki, Warmia i Mazury. Poznań: Instytut
Zachodni. Sezneva, O. (2007). “We have never been German”: The economy of digging in Russian
Kaliningrad. In C. Calhoun & R. Sennett (Eds.), Practicing culture (pp. 13–34). London:
Routledge. Sezneva, O. (2010). Modalities of self-understanding, identification and representation in
the post-1991 Kaliningrad: A critical view. In S. Berger (Ed.), Kaliningrad in Europe: Per-
spectives from inside and outside. Lüneburg: Nordost-Institut. Sezneva, O. (2013). Architecture of descent: Historical reconstructions and the politics
of belonging in Kaliningrad, the former Königsberg. Journal of Urban History, 39(4),
767–787. https://doi.org/10.1177/0096144212470095 Siegień, P. (2016). How much Königsberg is in Kaliningrad? New Eastern Europe, 2016(5),
115–122. Traba, R. (2005). Wschodniopruskość: Tożsamość regionalna i narodowa w kulturze poli-
tycznej Niemiec. Poznań: Poznańskie Towarzystwo Przyjaciół Nauk. Wiśniewska, I., Domańska, M., Strzelecki, J., Żochowski, P., Wilk, A., & Menkiszak, M. (2016). Kaliningrad Oblast 2016: The society, economy and army. Warsaw: Centre for
Eastern Studies. Андрей Клемешев: Учебник “История Западной России” должен быть заменён. (2017, Octo-
ber 10). Retrieved from http://vesti-kaliningrad.ru/a-klemeshev-o-novom-uchebnike-ob-
istorii/ Антон Алиханов открыл памятный знак павшим при Гросс-Егерсдорфе. (2017, August 30). Retrieved July 20, 2018, from http://ruwest.ru/news/74444/ Бродерзен, П. (2005). “Позови меня тихо по имени…”: Кампания переименований в Кали-
нинградской области в 1946-1950 гг. в контексте калининградско-московских отношений
послевоенного времени. In Балтийский регион в истории России и Европы: Материалы
международной научной конференции: Калининград, 29—30 октября 2004 г. Калинин-
град. Выползов, A. (2016). За “кёнигсбергизацией” Калининграда стоят США. REGNUM. Retrie-
ved July 20, 2018, from https://regnum.ru/news/polit/2160463.html Дементьев, И. О. (2014). “Что я могу знать?”: Формирование дискурсов о прошлом Кали-
нинградской области в советский период (конец 1940-х–1980-е годы). Люди и тексты:
Исторический альманах, 2014, 175–218. Bibliography Page 13 of 17 Page 13 of 17 “Если не станем противостоять в Калининграде, то начнём терять Россию по кусочкам”. (2017). Retrieved July 20, 2018, from http://newsbalt.ru/analytics/2017/07/esli-ne-sta-
nem-protivostoyat-v-kaliningrade-to-nachnem-teryat-rossiya-po-kusochkam/ “История Западной России” способствует росту сепаратизма в Калининграде. (2017). Re-
trieved July 20, 2018, from https://regnum.ru/news/2298605.html Клемешев, А. П., Костяшов, Ю. В., & Федоров, Г. М. (2007). История Западной России: Учеб-
ник 10-11 классы. Москва: ЗАО “ОЛМА Медиа Групп”. Колесников, А. (2015, June). Российская идеология после Крыма: Пределы эффективности
и мобилизации. Retrieved July 20, 2018, from http://carnegieendowment.org/files/
CMC_Brief_Kolesnikov_June2015_Rus.pdf Корвин, А. (2007). История Западной России. Retrieved July 20, 2018, from http://samlib. ru/k/korwin_a/zxcds.shtml Костяшов, Ю. В. (1996). Заселение Калининградской области после Второй мировой войны. In Гуманитарная наука в России: Соросовские лауреаты (Vol. 2, pp. 82–88). Москва:
Международный Научный Фонд. Костяшов, Ю. В. (2003). Изгнание прусского духа: Как формировалось историческое созна-
ние населения Калининградской области в послевоенные годы. Калининград: Издатель-
ство Калининградского гос. университета. Костяшов, Ю. В. (2009). Секретная история Калининградской области. Калининград: Терра
Балтика. Кретинин, Г. В. (2007a). История Западной России: Учебник 6-7 классы. Москва: ЗАО
“ОЛМА Медиа Групп”. Кретинин, Г. В. (2007b). История Западной России: Учебник 8-9 классы. Москва: ЗАО
“ОЛМА Медиа Групп”. Кретинин, Г. В., & Строганова, Н. А. (2006). Программа общего образования по истории
края. Калининград: РГУ им. И. Канта. Путин, В. (2012a, February 27). Россия и меняющийся мир. Московские новости. Retrieved
July 20, 2018, from http://www.mn.ru/politics/78738 Путин, В. (2012b, January 23). Россия: Национальный вопрос. Независимая газета. Retrie-
ved July 20, 2018, from http://www.ng.ru/politics/2012-01-23/1_national.html Путин, В. (2012c, January 16). Россия сосредотачивается — вызовы, на которые мы должны
ответить. Известия. Retrieved July 20, 2018, from http://izvestia.ru/news/511884 Сологубов, А. М. (2012). Переселенец как Homo Scientis: Эпистемологический аспект осво-
ения Калининградской области. Вестник БФУ им. И. Канта, 2012(6), 39–46. Сумарокова, А. (2017, August 19). “Ген.-фельдмарш. Румянцев”: В Калининграде появи-
лись новая улица и памятник. Retrieved July 20, 2018, from https://www.newkali-
ningrad.ru/news/community/14598360-gen-feldmarsh-rumyantsev-v-kaliningrade-
poyavilis-novaya-ulitsa-i-pamyatnik.html Хоппе, Б. (2005). Кёнигсберг/Калининград в ХХ в.—фокус современной европейской исто-
рии. In В. И. Гальцов (Ed.), Балтийский регион в истории России и Европы (pp. 190–
199). Калининград: Издательство РГУ им. И. Канта. Шабаева, Т. (2015, April 8). Родная память и чужое наследие. Литературная газета,
2015(14). Retrieved July 20, 2018, from http://www.lgz.ru/article/-14-6504-8-04-2015/
rodnaya-pamyat-i-chuzhoe-nasledie/ Шендерюк, М. Г. (n.d.). Социальный портрет переселенца. Retrieved from http://gako. name/index.php?publ132&razd207 Page 14 of 17 Bibliography (Transliteration) Bibliography (Transliteration) Andreı˘ Klemeshev: Uchebnik “Istoriia Zapadnoı˘ Rossii” dolzhen byt’ zamenën. (2017, Oc-
tober 10). Retrieved from http://vesti-kaliningrad.ru/a-klemeshev-o-novom-uchebnike-
ob-istorii/ Anton Alikhanov otkryl pamiatnyı˘ znak pavshim pri Gross-Egersdorfe. (2017, August 30). Retrieved July 20, 2018, from http://ruwest.ru/news/74444/ Broderzen, P. (2005). “Pozovi menia tikho po imeni…”: Kampaniia pereimenovaniı˘
v Kaliningradskoı˘ oblasti v 1946-1950 gg. v kontekste kaliningradsko-moskovskikh
otnosheniı˘ poslevoennogo vremeni. In Baltiı˘skiı˘ region v istorii Rossii i Evropy: Mate-
rialy mezhdunarodnoı˘ nauchnoı˘ konferentsii: Kaliningrad, 29—30 oktiabria 2004 g. Ka-
liningrad. Clark, C. (2009). Prusy: Powstanie i upadek: 1600-1947. Warszawa: Wydawnictwo Bel-
lona. Concept of the Foreign Policy of the Russian Federation. Approved by President
of the Russian Federation V. Putin on 12 February 2013. (2013). Retrieved July 20,
2018, from http://www.mid.ru/en/foreign_policy/official_documents/-/asset_publisher/
CptICkB6BZ29/content/id/122186 Dement’ev, I. O. (2014). “Chto ia mogu znat’?”: Formirovanie diskursov o proshlom
Kaliningradskoı˘ oblasti v sovetskiı˘ period (konets 1940-kh–1980-e gody). Liudi i teksty:
Istoricheskiı˘ al’manakh, 2014, 175–218. “Esli ne stanem protivostoiat’ v Kaliningrade, to nachnëm teriat; Rossiiu po kusochkam”. (2017). Retrieved July 20, 2018, from http://newsbalt.ru/analytics/2017/07/esli-ne-sta-
nem-protivostoyat-v-kaliningrade-to-nachnem-teryat-rossiya-po-kusochkam/ Heller, M. (1988). Cogs in the wheel: The formation of Soviet man. New York, NY: Alfred
A. Knopf. “Istoriia Zapadnoı˘ Rossii” sposobstvuet rostu separatizma v Kaliningrade. (2017). Retrie-
ved July 20, 2018, from https://regnum.ru/news/2298605.html Khoppe, B. (2005). Kënigsberg/Kaliningrad v XX v.—fokus sovremennoı˘ evropeı˘skoı˘ isto-
rii. In V. I. Gal’tsov (Ed.), Baltiı˘skiı˘ region v istorii Rossii i Evropy (pp. 190–199). Kalinin-
grad: Izdatel’stvo RGU im. I. Kanta. Klemeshev, A. P., Kostiashov, I. V., & Fedorov, G. M. (2007). Istoriia Zapadnoı˘ Rossii:
Uchebnik 10-11 klassy. Moskva: ZAO “OLMA Media Grupp”. Kolesnikov, A. (2015, June). Rossiı˘skaia ideologiia posle Kryma: Predely ėffektivnosti i
mobilizatsii. Retrieved July 20, 2018, from http://carnegieendowment.org/files/CMC_
Brief_Kolesnikov_June2015_Rus.pdf Korvin, A. (2007). Istoriia Zapadnoı˘ Rossii. Retrieved July 20, 2018, from http://samlib. ru/k/korwin_a/zxcds.shtml Kossert, A. (2009). Prusy Wschodnie: Historia i mit. Warszawa: Wydawnictwo Naukowe
Scholar. Kostiashov, I. V. (1996). Zaselenie Kaliningradskoı˘ oblasti posle Vtoroı˘ mirovoı˘ voı˘ny. In
Gumanitarnaia nauka v Rossii: Sorosovskie laureaty (Vol. 2, pp. 82–88). Moskva:
Mezhdunarodnyı˘ Nauchnyı˘ Fond. Kostiashov, I. V. (2003). Izgnanie prusskogo dukha: Kak formirovalos’ istoricheskoe soz-
nanie naseleniia Kaliningradskoı˘ oblasti v poslevoennye gody. Kaliningrad: Izdatel’stvo
Kaliningradskogo gos. universiteta. Page 15 of 17 Page 15 of 17 Kostiashov, I. V. (2009). Sekretnaia istoriia Kaliningradskoı˘ oblasti. Kaliningrad: Terra Balti-
ka. Kretinin, G. V. (2007a). Istoriia Zapadnoı˘ Rossii: Uchebnik 6-7 klassy. Moskva: ZAO
“OLMA Media Grupp”. Kretinin, G. V. (2007b). Istoriia Zapadnoı˘ Rossii: Uchebnik 8-9 klassy. Moskva: ZAO
“OLMA Media Grupp”. Kretinin, G. Bibliography (Transliteration) V., & Stroganova, N. A. (2006). Programma obshchego obrazovaniia po istorii
kraia. Kaliningrad: RGU im. I. Kanta. Marciniak, W. (2001). Rozgrabione imperium: Upadek Związku Sowieckiego i powstanie
Federacji Rosyjskiej. Kraków: Wydawnictwo Arcana. Putin, V. (2012a, February 27). Rossiia i meniaiushchiı˘sia mir. Moskovskie novosti. Retrie-
ved July 20, 2018, from http://www.mn.ru/politics/78738 Putin, V. (2012b, January 23). Rossiia: Natsional’nyı˘ vopros. Nezavisimaia gazeta. Re-
trieved July 20, 2018, from http://www.ng.ru/politics/2012-01-23/1_national.html Putin, V. (2012c, January 16). Rossiia sosredotachivaetsia — vyzovy, na kotorye my dol-
zhny otvetit’. Izvestiia. Retrieved July 20, 2018, from http://izvestia.ru/news/511884 Rodkiewicz, W., & Rogoża, J. (2015). Potemkin conservatism: An ideological tool
of the Kremlin. Warszawa: Ośrodek Studiów Wschodnich. Rogoża, J. (2013). Adjustment in Putin’s state: Under the banner of conservatism. Retrieved
July
20,
2018,
from
https://www.osw.waw.pl/en/publikacje/analy-
ses/2013-12-18/adjustment-putins-state-under-banner-conservatism Sakson, A. (2011). Od Kłajpedy do Olsztyna: Współcześni mieszkańcy byłych Prus
Wschodnich: Kraj Kłajpedzki, Obwód Kaliningradzki, Warmia i Mazury. Poznań: Instytut
Zachodni. Sezneva, O. (2007). “We have never been German”: The economy of digging in Russian
Kaliningrad. In C. Calhoun & R. Sennett (Eds.), Practicing culture (pp. 13–34). London:
Routledge. Sezneva, O. (2010). Modalities of self-understanding, identification and representation in
the post-1991 Kaliningrad: A critical view. In S. Berger (Ed.), Kaliningrad in Europe: Per-
spectives from inside and outside. Lüneburg: Nordost-Institut. Sezneva, O. (2013). Architecture of descent: Historical reconstructions and the politics
of belonging in Kaliningrad, the former Königsberg. Journal of Urban History, 39(4),
767–787. https://doi.org/10.1177/0096144212470095 Shabaeva, T. (2015, April 8). Rodnaia pamia’ i chuzhoe nasledie. Literaturnaia gazeta,
2015(14). Retrieved July 20, 2018, from http://www.lgz.ru/article/-14-6504-8-04-2015/
rodnaya-pamyat-i-chuzhoe-nasledie/ Shenderiuk, M. G. (n.d.). Sotsial’nyı˘ portret pereselentsa. Retrieved from http://gako. name/index.php?publ132&razd207 Siegień, P. (2016). How much Königsberg is in Kaliningrad? New Eastern Europe, 2016(5),
115–122. Sologubov, A. M. (2012). Pereselenets kak Homo Scientis: Ėpistemologicheskiı˘ aspekt
osvoeniia Kaliningradskoı˘ oblasti. Vestnik BFU im. I. Kanta, 2012(6), 39–46. Sumarokova, A. (2017, August 19). “Gen.-fel’dmarsh. Rumiantsev”: V Kaliningrade
poiavilas’ novaia ulitsa i pamiatnik. Retrieved July 20, 2018, from https://www.new-
kaliningrad.ru/news/community/14598360-gen-feldmarsh-rumyantsev-v-kaliningrade-
poyavilis-novaya-ulitsa-i-pamyatnik.html Page 16 of 17 Traba, R. (2005). Wschodniopruskość: Tożsamość regionalna i narodowa w kulturze poli-
tycznej Niemiec. Poznań: Poznańskie Towarzystwo Przyjaciół Nauk. Vypolzov, A. (2016). Za “kënigsbergizatsieı˘” Kaliningrada stoiat SShA. REGNUM. Retrie-
ved July 20, 2018, from https://regnum.ru/news/polit/2160463.html Wiśniewska, I., Domańska, M., Strzelecki, J., Żochowski, P., Wilk, A., & Menkiszak, M. (2016). Kaliningrad Oblast 2016: The society, economy and army. Warsaw: Centre for
Eastern Studies. Page 17 of 17
|
https://openalex.org/W4384931624
|
https://journal.unnes.ac.id/nju/index.php/sji/article/download/43438/pdf
|
English
| null |
Capital Optical Character Recognition Using Neural Network Based on Gaussian Filter
|
Scientific Journal of Informatics/Scientific journal of informatics
| 2,023
|
cc-by
| 6,297
|
Abstract. Purpose: As digital technology advances, society needs to convert physical text into digital text. There are now many
methods available for doing this. One of them is OCR (Optical Character Recognition), which can scan images [1]–[4]
containing writing and turn them into digital text, making it easier to copy written text from an image. Text recognition
in images is complex due to variations in text size, color, font, orientation, background, and lighting conditions. g
p
g
g
g
Methods: The technique of text recognition or optical character recognition (OCR) in images can be done using several
methods, one of which is a neural network or artificial neural network. The artificial neural network method can help
a computer make intelligent decisions with limited human assistance. Intelligent decisions can be made because the
neural network can learn and model the relationship between nonlinear and complex input and output data. In this
research, the scaled conjugated gradient is applied for optimization. SCG is very effective in finding the minimum
value of a complex function, but it takes longer than some other optimization algorithms. Result/Findings: The dataset
used is an image with a size of 28 x 28 which is changed in dimension to 784 x 1. This research uses 4000 epochs and
obtained the best validation result at epoch 3506 with a value of 0.0087446. Results: From the statistical test results, the effect of perceived usefulness on ease of use has the highest level of
influence, obtaining a test value of 3.6. Furthermore, the effect of the attitude towards using on the behavioral intention
to use has the lowest level of influence, which obtained a test value of 1.2. Novelty: In this article, Gaussian filter is used as feature extraction to improve yield. Character detection results using
a Gaussian filter are known to be almost 10% higher than those using only a neural network. The result with the Neural
Network alone is 82.2%, while the Neural Network-Gaussian Filter produces 92.1%. Keywords: Digital test, Neural network, Optical character recognition, Test recognition
Received March 2023 / Revised May 2023 / Accepted June 2023 This work is licensed under a Creative Commons Attribution 4.0 International License. Capital Optical Character Recognition Using Neural Network Based
on Gaussian Filter Erna Zuni Astuti1*, Christy Atika Sari2, Mutiara Salsabila3, Hendra Sutrisno4, Eko Hari
Rachmawanto5, Mohamed Doheir6
1,2,3,4,5Department of Informatics Engineering, Faculty of Computer Science, Universitas Dian Nuswantoro, Indonesia
6Department of Computer Science, University Geomatrika, Malaysia Scientific Journal of Informatics
Vol. 10, No. 3, Aug 2023
http://journal.unnes.ac.id/nju/index.php/sji p-ISSN 2407-7658 e-ISSN 2460-0040 *Corresponding author.
Email addresses: erna.zuni.astuti@dsn.dinus.ac.id (Astuti)
DOI: 10.15294/sji.v10i3.43438 Scientific Journal of Informatics, Vol. 10, No. 3, Aug 2023 | 261 INTRODUCTION OCR (Optical Character Recognition)
can use neural network or artificial neural network methods to improve accuracy in identifying letters in
images or documents. Artificial neural networks can help OCR by processing the data given and finding
patterns that can be used to identify those letters [10], [16], [27]. Using a neural network, OCR can be more
accurate in identifying letters that are difficult to distinguish, such as the letters "D" and "O." The advantage
of artificial neural networks is their ability to recognize patterns from previously taught images. In previous
research, the same topic was discussed and can be used as a reference for comparison or to add further
discussion about the topic being researched. In this research, several journals discussing the same topic,
text recognition, were used as a comparison. The following are the journals by Khandokar [15] in 2021
using Convolutional Neural Network (CNN) to recognize the characters based on NIST dataset. This
experiment yield 92.91% using 80:20 for training and testing data form 1000 image, while accuracy only
65.32% using 200 datasets. Using segmentation, clipping, pre-processing, and feature extraction, the results
of this study might be optimized by combining or replacing extraction features. Another research by
Susanto [28] in 2021 has Javanese alphabet classification using simple machine learning by K-Nearest
Neighbor (KNN) and optimized using cropping, median filtering, otsu thresholding and HOG feature
extraction. The highest accuracy is 98,5% in K=1. The concept of artificial neural networks began in the 1940s and 1950s, when researchers began to
understand how the brain processes information by studying the structure and function of the brain. In 1943,
Warren McCulloch and Walter Pitts published a paper proposing that neurons in the brain could be
represented as simple logic gates that accept inputs and produce outputs based on a set of rules [7], [18],
[21], [24], [29]. Announced. This idea led Frank Rosenblatt to develop the first artificial neural network,
the so-called perceptron, in the 1950s. A perceptron was a simple neural network consisting of a single
layer of artificial neurons that could be trained to recognize patterns in data. However, perceptrons were
limited in their ability to recognize only linearly separable patterns, limiting their ability to solve more
complex problems. In the 1980s, the introduction of backpropagation, a learning algorithm that adjusts the
weights and biases of neural networks to improve their performance, revived the development of artificial
neural networks [30]. INTRODUCTION As digital technology advances, society needs printed text such as books, journals, and documents to be
converted into digital form in order to increase efficiency in automatically converting documents into digital
files. Text recognition in images is complex due to variations in text size, color, font, orientation,
background, low image resolution, and lighting conditions [1]–[4]. In addition, similar image shapes greatly
affect text recognition accuracy. The problem that arises is how to transfer the printed text to digital text
form. A system to recognize text in images [5]–[10] is known as an optical character recognition (OCR)
system. OCR was developed to address this problem. Optical character recognition (OCR) is a technology
that allows computers to read text contained in images or printed documents [4], [11]. OCR is very useful
for converting image documents or text images into editable text. OCR can be used to read text contained
in images, such as in photos, books, or documents [11], [12]. OCR works by scanning an image or document
and recognizing the letters contained in the image. After the letters are recognized, OCR uses complex
algorithms to convert the letters into digital text that can be processed by a computer. This is done by
comparing the recognized letters with a database of letters already recognized by OCR [1], [13]–[15]. If
OCR cannot find a letter that matches its known dataset, it will try to guess the letter with a shape similar
with its database [9], [16], [17]. *Corresponding author. Email addresses: erna.zuni.astuti@dsn.dinus.ac.id (Astuti)
DOI: 10.15294/sji.v10i3.43438 Scientific Journal of Informatics, Vol. 10, No. 3, Aug 2023 | 261 This research discusses an OCR-based system using the neural network or artificial neural network method. A neural network or artificial neural network is a set of algorithms used to imitate the workings of the
human nervous system. Artificial neural networks [18] are one of the deep learning algorithms, which
become more intelligent over time. Artificial neural networks [4], [13], [15], [19]–[22]] can learn on their
own through experience and can become more accurate as more data is provided. A neural network is a
system that processes information by imitating the workings of the human brain, allowing a computer to
understand various things and make decisions like a human think [7], [8], [23], [24]. Neural networks can
be used for a variety of tasks, such as pattern recognition [25], [26]. INTRODUCTION This has led to the creation of multi-layer neural networks (also known as deep
learning networks) that can learn and recognize more complex patterns in data. Artificial neural networks
are used today in a variety of applications, including image and speech recognition, natural language
processing, and machine learning. It has also played a key role in the development of self-driving cars and
other emerging technologies. The main purpose of this journal is to create a text recognition application
that can facilitate the process of converting printed images or documents into editable digital text. The text
recognition application can be useful for converting image documents or text images into editable text. The
text recognition application can also be used to read text contained in images, such as book photos or
documents. This can help facilitate document archiving. METHODS Then, segmentation is also carried out to focus on specific parts. Preprocessing and segmentation are important steps in the OCR process as they help improve the quality
of the input photos and focus on the specific area of interest, which can help improve the accuracy of the
OCR system [5], [23], [35], [36]. Preprocessing can involve steps such as image resizing, noise removal,
and image enhancement, while segmentation involves dividing the image into smaller regions or segments. These steps can help the OCR system to enhanced accurately identify and recognize the text in the image. The dataset used in this research was taken from Kaggle, which contains 372038 characters image a set of
26 capital character (A to Z) printed in the System font. Each letter image is represented by 28 x 28 pixels
and have a black background that has been resized to 784 x 1, where each pixel can be either on (represented
by a '1') or off (represented by a '0'). The process involves scanning each pixel in the enhanced letter image
from top to bottom and left to right to locate the capital letter printed on the paper. It is assumed that the
letters are clearly separated from each other (capital letter). In this research, we use the neural network
method. A neural network or artificial neural network is a system consisting of many units called "neurons"
that work together to solve problems that the system will face [12], [37]. The way a neural network works
is that it receives input data to be processed. This input is processed by the neurons in the network. Each
neuron calculates its output value using a predetermined function [38]–[40]. Figure 1. Sample data collection
Figure 2. Flowchart proposed method Figure 2. Flowchart proposed method In this research, a Neural Network will be used. However, before training, preprocessing must be done to
make the image clearer and cleaner. Then, segmentation is also carried out to focus on specific parts. Preprocessing and segmentation are important steps in the OCR process as they help improve the quality
of the input photos and focus on the specific area of interest, which can help improve the accuracy of the
OCR system [5], [23], [35], [36]. Preprocessing can involve steps such as image resizing, noise removal,
and image enhancement, while segmentation involves dividing the image into smaller regions or segments. METHODS Research on text recognition in images or OCR (Optical Character Recognition) is very important because
it can help in processing and accessing the information contained in photos by converting it into machine-
readable text [14], [31]–[34]. OCR can be used in various fields such as education, business, and industry. In the education field, OCR can help in processing and storing documents contained in images such as
textbooks, lecture notes, and other documents. OCR can also help in accessing documents written in
unusual letters or in a foreign language that is not known to someone. In the business field, OCR can help
in processing and storing documents contained in images such as invoices, financial reports, and other
documents. OCR can also help in processing and storing data contained in photos such as names, addresses,
and phone numbers, which can be used for marketing or data analysis purposes. In the industrial field, OCR
can help in processing and storing documents contained in images, such as product specifications, quality
reports, and other documents. OCR can also help in processing and storing data contained in images, such
as serial numbers, production dates, and production locations, which can be used for product maintenance
and care purposes. This research on text recognition in images is very important because it can help in 262 | Scientific Journal of Informatics, Vol. 10, No. 3, Aug 2023 processing and accessing the information contained in photos, which can be useful in various fields. Text
recognition in photos is a very complex matter due to the many variations. For example, different colors,
sizes, fonts, letter sizes, background images, and lighting conditions. Therefore, it is very difficult to
achieve high accuracy even though reCAPTCHA is currently still collecting datasets to improve its
program. Therefore, in this research, only capital text is used as shown in Figure 1. The objects used in this
research are a document and an image containing capital letter text. The document and photos will be input
into the system to read and recognize the text contained within it. The objects of this research were collected
from various sources, such as books, letters, and personal documents. from various sources, such as books, letters, and personal documents. Figure 1. Sample data collection
Figure 2. Flowchart proposed method
In this research, a Neural Network will be used. However, before training, preprocessing must be done to
make the image clearer and cleaner. METHODS The number of units in the input layer is determined by the dimensions of the
input data. For example, if the input data is a 2D image with 28 x 28 pixels, then the input layer would have
28 x 28 = 784 units. If the input data is a 1D time series with 10 times steps, then the input layer would
have 10 units. The input layer does not have any internal weights or biases, as it does not perform any
computations. It simply serves as a conduit for the input data to flow through the network to the subsequent
layers. And then there is also a hidden layer. A hidden layer in a neural network is a layer of neurons that
is not visible to the input data or the output predictions of the network. Hidden layers are located between
the input layer and the output layer in a neural network and are responsible for extracting features from the
raw input data and transforming it into a more useful representation for the output layer to process. The
number of hidden layers and the number of neurons in each hidden layer can vary greatly depending on the
complexity of the problem being solved. In general, a deeper neural network with more hidden layers can
learn more complex relationships in the data, but it also requires more computational resources and can be
more prone to overfitting. Each hidden layer in a neural network consists of a set of neurons, each of which
receives input from the neurons in the previous layer, applies a nonlinear activation function, and passes
the output to the neurons in the next layer. The activation function is a mathematical function that
determines the output of a neuron given its input. Common activation functions include sigmoid, tanh, and
ReLU (Rectified Linear Unit). Figure 3. Illustration of neural network [4][8][30] Figure 3. Illustration of neural network [4][8][30] Figure 3. Illustration of neural network [4][8][30] Figure 3. Illustration of neural network [4][8][30] The weights and biases of the connections between the neurons in the hidden layers are learned during the
training process of the neural network using an optimization algorithm, such as stochastic gradient descent. The learning process involves adjusting the weights and biases in order to minimize the error between the
predicted output and the ground truth label for a given input. METHODS These steps can help the OCR system to enhanced accurately identify and recognize the text in the image. The dataset used in this research was taken from Kaggle, which contains 372038 characters image a set of
26 capital character (A to Z) printed in the System font. Each letter image is represented by 28 x 28 pixels
and have a black background that has been resized to 784 x 1, where each pixel can be either on (represented
by a '1') or off (represented by a '0'). The process involves scanning each pixel in the enhanced letter image
from top to bottom and left to right to locate the capital letter printed on the paper. It is assumed that the
letters are clearly separated from each other (capital letter). In this research, we use the neural network
method. A neural network or artificial neural network is a system consisting of many units called "neurons"
that work together to solve problems that the system will face [12], [37]. The way a neural network works
is that it receives input data to be processed. This input is processed by the neurons in the network. Each
neuron calculates its output value using a predetermined function [38]–[40]. The output of each neuron is summed and sent to the neurons in the next layer. This process continues until
the data reaches the output layer, where the final output of the network is produced. During the learning
process, neural networks can learn patterns in data. For example, there may be specific features on an object
to be recognized. For example, the letter "A" has characteristics such as a sloping line, a horizontal line, Scientific Journal of Informatics, Vol. 10, No. 3, Aug 2023 | 263 and so on. During the training process, the neural network will try to identify these characteristics. On a
neural network there is such a thing as input layer. The input layer of a neural network is the layer that
receives the input data. It is the first layer of the network and does not perform any computations or
transformations on the data. Its primary role is to pass the input data through the network to the subsequent
layers for processing. The input layer is composed of units in Figure 3, also known as neurons or nodes,
that represent the input data. METHODS In this research we used 65 hidden layers. Having 65 hidden layers in a neural network would be considered a very deep neural network. Deep neural
networks, which have many hidden layers able to learn and model very complex relationships in the data. However, they also require a lot of computational resources to train and can be prone to overfitting, which
means that they perform well on the training data but do not generalize well to unseen data. There are
several factors to consider when deciding how many hidden layers to include in a neural network One factor is the complexity of the problem being solved. A deeper neural network may be necessary for
more complex tasks, but it may not be necessary for simpler tasks. Another factor is the amount of data
available for training. A deeper neural network may be able to learn more complex relationships in the data,
but it also requires more data to learn from in order to avoid overfitting. In general, it is a good idea to start
with a relatively shallow neural network and gradually increase the depth as needed, rather than starting 264 | Scientific Journal of Informatics, Vol. 10, No. 3, Aug 2023 with a very deep neural network. This allows you to determine the optimal number of hidden layers for
your specific problem, while minimizing the risk of overfitting. and the last is output layer. In a neural
network, the output layer is the final layer of neurons in the network. It is responsible for producing the
output of the neural network based on the inputs it receives from the previous layers. The output layer
receives input from the other layers in the network, processes this input using the weights and biases of the
neurons, and produces the final output of the network. The output of the output layer is often used to make
a prediction or classification based on the input data. The number of neurons in the output layer is typically
determined by the number of classes that the network is trying to predict. For example, if the neural network
is trying to classify images into one of 10 different categories, the output layer would have 10 neurons, each
corresponding to a different class. The output of each neuron in the output layer would be a probability,
indicating the likelihood that the input image belongs to that class. MSE = (1/n) ∑(Yi - Yi')^2 (1) Scientific Journal of Informatics, Vol. 10, No. 3, Aug 2023 | 265 METHODS The class with the highest probability
would be the one that the network predicts. In some cases, the output layer may also have a continuous
output, such as a real number, rather than a probability. This can be useful for tasks such as regression,
where the goal is to predict a continuous output rather than a discrete class. Overall, the output layer is an
important part of a neural network, as it produces the final output of the network based on the input it
receives from the other layers. This measures how well your regression model to predict the actual output
for new data that was not used in the modeling. If your regression model is good, then it will predict the
actual output with a low level of error. Figure 6 shows the best validation result, which is 0.0087446 at
epoch 3506. Mean squared error (MSE) is one matrix that can be used to evaluate the performance of a
neural network validation. MSE is calculated by taking the difference between the desired output value and
the actual output value produced by the model, then multiplying the difference by 2, and then summing all
the results. Then, the sum is divided by the number of data used for validation. MSE is used to measure the
difference between the predicted output and the actual output, and a lower MSE indicates a better model. To calculate MSE, you first need to calculate the difference between the predicted value and the actual
value for each data point, square the difference, and then sum all the squared differences. Finally, divide
the sum by the number of data points to get the average squared error. The lower the MSE, the better the
model is at predicting the actual output. The smaller the MSE value, the better the model is considered at
predicting the desired output. However, it is important to note that MSE is not always the most appropriate
metric for every case. If the data used has very large values, MSE may not give an accurate picture of the
model's performance as in (1), where n is the number of data points used for validation, ∑ is the sum symbol,
indicating that the values should be added together, Predicted value is the value predicted by model, Actual
value is the true value of the output. RESULTS AND DISCUSSIONS Before creating the neural network, we import the existing dataset. In this case, we create 65 hidden layers
in the neural network. Before training the input from the dataset, we first normalize it using a custom
function. This function will change the background of the images in the dataset to white. Then we normalize
the target to be trained, resulting in the transpose of the normalized target. Here, we use 4000 epochs or
iterations. The training process will take approximately 2-4 hours and will result in the following
performance. Based on Figure 4 and Figure 5, the value of R for Training, Validation, and Test is 0.89136
or 89.1%, 0.87411 or 87.4%, and 0.86548, respectively. Scientific Journal of Informatics, Vol. 10, No. 3, Aug 2023 | 265 Figure 4. Regression training neural network result Figure 4. Regression training neural network result Inputting an image is the first process carried out. In this process, all input images are read by the system. The image is then processed in the next step, which is pre-processing. After creating an artificial neural
network, the next step is to pre-process the input image. Pre-processing is the process of cleaning and
preparing the data before it is used in training. Pre-processing is the process of preparing the image for
further analysis by cleaning, enhancing, and adjusting the image so that it can be accurately processed by
the OCR system. 266 | Scientific Journal of Informatics, Vol. 10, No. 3, Aug 2023
the OCR system. Figure 5. Validation performance Figure 5. Validation performance Figure 5. Validation performance 266 | Scientific Journal of Informatics, Vol. 10, No. 3, Aug 2023 266 | Scientific Journal of Informatics, Vol. 10, No. 3, Aug 2023 This may involve tasks such as removing noise or blur, correcting distortions, and adjusting the image
contrast. Pre-processing is an important step in the OCR process because it helps to improve the accuracy
and efficiency of the OCR system. The first step is to add a Gaussian filter, which is used to smooth the
indentations in the character and improve the accuracy of the output. Then, the image is converted from
grayscale to binary. Next, a cropping step is performed on the input image. Cropping is done to discard
unnecessary parts by finding rows and columns that only contain a value of 0. RESULTS AND DISCUSSIONS By applying a Gaussian filter to the input image before processing it with
the OCR system, the quality of the image can be improved, which can help improve the accuracy of the
OCR system. The testing using Gaussian filter show an average accuracy of 92.1%, with the highest
accuracy at 100% and the lowest at 60%. This is because the objects being predicted are too close together,
so when segmenting the letters, the letters that are touching are only read as one. Images will be difficult to
read if the background of the image is not clean, dark, and uneven. Therefore, applying a Gaussian filter in
the preprocessing stage is important because it removes noise from the image, which improves the accuracy
of text recognition and the quality of the image. By improving the quality of the input image, the OCR
system is more likely to accurately recognize the text in the image. RESULTS AND DISCUSSIONS The next step is segmentation,
which is used to separate each character or letter by storing two indexes for the beginning and end in an
array. Segmenting the parts or each letter is useful to make the text recognition more focused and get more
specific information from the object. Then, cropping of the image containing the word into each letter will
be performed using the two initial indexes and the saved end index previously. After this step, the following
will be produced. After marking the parts of the letters that will be matched with the trained network,
cropping of the image containing the word will be performed into individual letters using the previously
stored initial and final indexes. Here, will use images specified in the object section. This study uses 15
testing data to test the model that has been created. This study will compare the results of testing using a
gaussian filter during preprocessing and without using a gaussian filter. Table 1. Comparison accuracy
Original Character
Neural Network
Gussian Filter + Neural Network
Recognized
Accuracy
Recognized
Accuracy
PEREMPUAN
JJAWXH
0%
PEREMOUAN
88.8%
KARTU
KARTU
100%
KARTU
100%
MAHASISWA
UAHASRSWA
77.7%
MAHASBSWA
88.8%
SEMENTARA
SEMENTARA
100%
SEMENTARA
100%
CITRA
CRTRA
75%
CITRA
100%
DIGITAL
DRGRTAL
71.4%
DIGBTAL
85.7%
PENGOLAHAN
PENGOLAHAN
100%
PENGOLAHAN
100%
KESEHATAN
KESEHAAN
88.8%
KESEHAAN
88.8%
PESERTA
PESERTA
100%
PESERTA
100%
NEGERI
NEGERB
83,3%
NEGERB
83.3%
SEKOLAH
SEKOLAH
100%
SEKOLAH
100%
KEBUDAYAAN
KEBUDAYAAR
90%
KEBUDAYAAN
100%
REPUBLIK
REPUBLBK
87,5%
REPUBLBK
87.5%
NUSWANTORO
NUSWANTORO
100%
NUSWANTORO
100%
UJIAN
UJIB
60%
UJIB
60%
Average
82,2%
Average
92,1% Based on Table 1, the testing results of the test show an average accuracy of 82.2, with the highest accuracy
at 100% and the lowest at 0%. This is because the image is not given a Gaussian filter, resulting in noise
being created on the background when binarized, which reduces the accuracy of text recognition. Without
using a Gaussian filter, noise cannot be removed, and the quality of the photos is reduced, which also
reduces the accuracy of text recognition and can cause errors in text recognition. Gaussian filters are
commonly used in image processing to smooth and blur photos, and they can be used to reduce noise and
improve the quality of photos. Scientific Journal of Informatics, Vol. 10, No. 3, Aug 2023 | 267 REFERENCES [1]
V. V Mainkar, J. A. Katkar, A. B. Upade, and P. R. Pednekar, “Handwritten Character Recognition
to Obtain Editable Text,” 2020 Int. Conf. Electron. Sustain. Commun. Syst., pp. 599–602, 2020. [2]
Y. Wang, W. Xiao, and S. Li, “Offline Handwritten Text Recognition Using Deep Learning: A
Review,” J. Phys. Conf. Ser., vol. 1848, no. 1, p. 012015, Apr. 2021, doi: 10.1088/1742-
6596/1848/1/012015. [3]
M. Wang, S. Niu, and Z. Gao, “A Novel Scene Text Recognition Method Based on Deep Learning,”
Comput. Mater. Contin., vol. 60, no. 2, pp. 781–794, 2019, doi: 10.32604/cmc.2019.05595. [4]
R. Parthiban, R. Ezhilarasi, and D. Saravanan, “Optical Character Recognition for English
Handwritten Text Using Recurrent Neural Network,” in 2020 International Conference on System,
Computation,
Automation
and
Networking
(ICSCAN),
Jul. 2020,
pp. 1–5. doi:
10.1109/ICSCAN49426.2020.9262379. [5]
M. B. Bora, D. Daimary, K. Amitab, and D. Kandar, “Handwritten Character Recognition from
Images using CNN-ECOC,” Procedia Comput. Sci., vol. 167, pp. 2403–2409, 2020, doi:
10.1016/j.procs.2020.03.293. j p
[6]
Y. Wang, L. Dang, and J. Ren, “Forest fire image recognition based on convolutional neural
network,” J. Algorithm. Comput. Technol., vol. 13, p. 174830261988768, Jan. 2019, doi:
10.1177/1748302619887689. [7]
S. Aqab and M. Usman, “Handwriting Recognition using Artificial Intelligence Neural Network
and Image Processing,” Int. J. Adv. Comput. Sci. Appl., vol. 11, no. 7, 2020, doi:
10.14569/IJACSA.2020.0110719. [8]
W. Mo, X. Luo, Y. Zhong, and W. Jiang, “Image recognition using convolutional neural network
combined with ensemble learning algorithm,” J. Phys. Conf. Ser., vol. 1237, no. 2, p. 022026, Jun. 2019, doi: 10.1088/1742-6596/1237/2/022026. [9]
Y. B. Hamdan and A. Sathesh, “Deep Learning based Handwriting Recognition with Adversarial
Feature Deformation and Regularization,” J. Innov. Image Process., vol. 3, no. 4, pp. 367–376, Dec. 2021, doi: 10.36548/jiip.2021.4.008. [10]
Tapotosh Ghosh, M.-H.-Z. Abedin, H. Al Banna, N. Mumenin, and M. Abu Yousuf, “Performance
Analysis of State of the Art Convolutional Neural Network Architectures in Bangla Handwritten
Character Recognition,” Pattern Recognit. Image Anal., vol. 31, no. 1, pp. 60–71, Jan. 2021, doi:
10.1134/S1054661821010089. [11]
G. A. Robby, A. Tandra, I. Susanto, J. Harefa, and A. Chowanda, “Implementation of Optical
Character Recognition using Tesseract with the Javanese Script Target in Android Application,”
Procedia Comput. Sci., vol. 157, pp. 499–505, 2019, doi: 10.1016/j.procs.2019.09.006. [12]
E. A. Al-Zubaidi, M. M. Mijwil, and A. S. Alsaadi, “Two-Dimensional Optical Character
Recognition of Mouse Drawn in Turkish Capital Letters Using Multi-Layer Perceptron
Classification,” J. Southwest Jiaotong Univ., vol. 54, no. CONSLUSION This research was conducted on text recognition using the neural network method with and without adding
a Gaussian filter. From the results of this research and testing, it can be concluded that the system using a
Gaussian filter has a success rate of text recognition with an accuracy of 92.1%, while the system without
using a Gaussian filter has a success rate of text recognition with an accuracy of 82.2%. It can therefore be
concluded that text recognition using the neural network method assisted by a Gaussian filter will produce
a higher accuracy than text recognition without using a Gaussian filter. Gaussian filters can improve the Scientific Journal of Informatics, Vol. 10, No. 3, Aug 2023 | 267 quality of the input image by removing noise and smoothing the image, which can help improve the
accuracy of the OCR system. It is important to consider the use of preprocessing techniques such as
Gaussian filtering to improve the accuracy of OCR systems. For the future research, Neural Network may
be can implemented using Wiener filter or Adaptive Threshold to increase best pixel enhancement before
weighting and is expected to produce higher accuracy. REFERENCES 4, 2019, doi: 10.35741/issn.0258-
2724.54.4.4. [13]
H. M. Najadat, A. A. Alshboul, and A. F. Alabed, “Arabic Handwritten Characters Recognition
using Convolutional Neural Network,” in 2019 10th International Conference on Information and
Communication Systems (ICICS), Jun. 2019, pp. 147–151. doi: 10.1109/IACS.2019.8809122. [14]
A. Rao, A. Arpitha, C. Nayak, S. Meghana, S. Nayak, and S. S., “Exploring Deep Learning
Techniques for Kannada Handwritten Character Recognition: A Boon for Digitization,” vol. 29,
pp. 11078–11093, Jul. 2020. [15]
I. Khandokar, M. Hasan, F. Ernawan, S. Islam, and M. N. Kabir, “Handwritten character
recognition using convolutional neural network,” J. Phys. Conf. Ser., vol. 1918, no. 4, p. 042152,
Jun. 2021, doi: 10.1088/1742-6596/1918/4/042152. [16]
I. Uddin et al., “Benchmark Pashto Handwritten Character Dataset and Pashto Object Character
Recognition (OCR) Using Deep Neural Network with Rule Activation Function,” Complexity, vol. 2021, pp. 1–16, Mar. 2021, doi: 10.1155/2021/6669672. pp
[17]
Y. B. Hamdan and Sathish, “Construction of Statistical SVM based Recognition Model for
Handwritten Character Recognition,” J. Inf. Technol. Digit. World, vol. 3, no. 2, pp. 92–107, Jun. 268 | Scientific Journal of Informatics, Vol. 10, No. 3, Aug 2023 2021, doi: 10.36548/jitdw.2021.2.003. [18] [18]
M. H. KESİKOĞLU, S. Y. ÇİÇEKLİ, and T. KAYNAK, “THE IDENTIFICATION OF
SEASONAL COASTLINE CHANGES FROM LANDSAT 8 SATELLITE DATA USING
ARTIFICIAL NEURAL NETWORKS AND K-NEAREST NEIGHBOR,” Turkish J. Eng., vol. 4,
no. 1, pp. 47–56, Jan. 2020, doi: 10.31127/tuje.599359. pp
j
[19]
S. Ahlawat, A. Choudhary, A. Nayyar, S. Singh, and B. Yoon, “Improved Handwritten Digit
Recognition Using Convolutional Neural Networks (CNN),” Sensors, vol. 20, no. 12, p. 3344, Jun. 2020, doi: 10.3390/s20123344. [20]
A. K. Nugroho, I. Permadi, and M. Faturrahim, “Improvement Of Image Quality Using
Convolutional Neural Networks Method,” Sci. J. Informatics, vol. 9, no. 1, pp. 95–103, May 2022,
doi: 10.15294/sji.v9i1.30892. [21]
T. Q. Vinh, L. H. Duy, and N. T. Nhan, “Vietnamese handwritten character recognition using
convolutional neural network,” IAES Int. J. Artif. Intell., vol. 9, no. 2, p. 276, Jun. 2020, doi:
10.11591/ijai.v9.i2.pp276-281. [22]
N. B. Muppalaneni, “Handwritten Telugu Compound Character Prediction using Convolutional
Neural Network,” in 2020 International Conference on Emerging Trends in Information
Technology and Engineering (ic-ETITE), 2020, pp. 1–4. doi: 10.1109/ic-ETITE47903.2020.349. [23]
J. Liu and J. Hong, “Design of Handwritten Numeral Recognition System Based on BP Neural
Network,” J. Phys. Conf. Ser., vol. 2025, no. 1, p. 012016, Sep. 2021, doi: 10.1088/1742-
6596/2025/1/012016. [24]
A. Mohsin and M. REFERENCES Sadoon, “Developing an Arabic Handwritten Recognition System by Means of
Artificial Neural Network,” J. Eng. Appl. Sci., vol. 15, no. 1, pp. 1–3, Oct. 2019, doi:
10.36478/jeasci.2020.1.3. j
[25]
Y. Zhuang et al., “A Handwritten Chinese Character Recognition based on Convolutional Neural
Network and Median Filtering,” J. Phys. Conf. Ser., vol. 1820, no. 1, p. 012162, Mar. 2021, doi:
10.1088/1742-6596/1820/1/012162. [26]
R. B. Lincy and R. Gayathri, “Optimally configured convolutional neural network for Tamil
Handwritten Character Recognition by improved lion optimization model,” Multimed. Tools Appl.,
vol. 80, no. 4, pp. 5917–5943, Feb. 2021, doi: 10.1007/s11042-020-09771-z. pp
[27]
N. P. Sutramiani, N. Suciati, and D. Siahaan, “MAT-AGCA: Multi Augmentation Technique on
small dataset for Balinese character recognition using Convolutional Neural Network,” ICT
Express, vol. 7, no. 4, pp. 521–529, Dec. 2021, doi: 10.1016/j.icte.2021.04.005. [28]
A. Susanto, C. Atika Sari, I. U. W. Mulyono, and M. Doheir, “Histogram of Gradient in K-Nearest
Neighbor for Javanese Alphabet Classification,” Sci. J. Informatics, vol. 8, no. 2, pp. 289–296,
Nov. 2021, doi: 10.15294/sji.v8i2.30788. [29]
D. Beohar and A. Rasool, “Handwritten Digit Recognition of MNIST dataset using Deep Learning
state-of-the-art Artificial Neural Network (ANN) and Convolutional Neural Network (CNN),” in
2021 International Conference on Emerging Smart Computing and Informatics (ESCI), Mar. 2021,
pp. 542–548. doi: 10.1109/ESCI50559.2021.9396870. pp
[30]
I. Prihandi, I. Ranggadara, S. Dwiasnati, Y. S. Sari, and Suhendra, “Implementation of
Backpropagation Method for Identified Javanese Scripts,” J. Phys. Conf. Ser., vol. 1477, no. 3, p. 032020, Mar. 2020, doi: 10.1088/1742-6596/1477/3/032020. [31]
A. Hazra, P. Choudhary, S. Inunganbi, and M. Adhikari, “Bangla-Meitei Mayek scripts handwritten
character recognition using Convolutional Neural Network,” Appl. Intell., vol. 51, no. 4, pp. 2291–
2311, Apr. 2021, doi: 10.1007/s10489-020-01901-2. p
[32]
R. KARAKAYA and S. KAZAN, “Handwritten Digit Recognition Using Machine Learning,” Sak. Üniversitesi Fen Bilim. Enstitüsü Derg., vol. 25, no. 1, pp. 65–71, Feb. 2021, doi:
10.16984/saufenbilder.801684. [33]
M. Jain, G. Kaur, M. P. Quamar, and H. Gupta, “Handwritten Digit Recognition Using CNN,” in
2021 International Conference on Innovative Practices in Technology and Management (ICIPTM),
Feb. 2021, pp. 211–215. doi: 10.1109/ICIPTM52218.2021.9388351. pp
[34]
S. Ahlawat and A. Choudhary, “Hybrid CNN-SVM Classifier for Handwritten Digit Recognition
Procedia Comput. Sci., vol. 167, pp. 2554–2560, 2020, doi: 10.1016/j.procs.2020.03.309. [35]
S. P. Deore and A. Pravin, “Devanagari Handwritten Character Recognition using fine-tuned Deep
Convolutional Neural Network on trivial dataset,” Sādhanā, vol. 45, no. 1, p. 243, Dec. 2020, doi:
10.1007/s12046-020-01484-1. [36]
P. (Dr. . 270 | Scientific Journal of Informatics, Vol. 10, No. 3, Aug 2023 REFERENCES R. S. Anuj Bhardwaj, “HANDWRITTEN DEVANAGARI CHARACTER
RECOGNITION USING DEEP LEARNING - CONVOLUTIONAL NEURAL NETWORK Scientific Journal of Informatics, Vol. 10, No. 3, Aug 2023 | 269 (CNN) MODEL,” PalArch’s J. Archaeol. Egypt / Egyptol., vol. 17, no. 6 SE-, pp. 7965–7984, Dec. 2020, [Online]. Available: https://archives.palarch.nl/index.php/jae/article/view/2203 [37]
M. S. Alam, K.-C. Kwon, M. A. Alam, M. Y. Abbass, S. M. Imtiaz, and N. Kim, “Trajectory-Based
Air-Writing Recognition Using Deep Neural Network and Depth Sensor,” Sensors, vol. 20, no. 2,
p. 376, Jan. 2020, doi: 10.3390/s20020376. p
[38]
R. Dixit, R. Kushwah, and S. Pashine, “Handwritten Digit Recognition using Machine and Deep
Learning Algorithms,” Int. J. Comput. Appl., vol. 176, no. 42, pp. 27–33, Jul. 2020, doi:
10.5120/ijca2020920550. [39]
A. Anton, N. F. Nissa, A. Janiati, N. Cahya, and P. Astuti, “Application of Deep Learning Using
Convolutional Neural Network (CNN) Method For Women’s Skin Classification,” Sci. J. Informatics, vol. 8, no. 1, pp. 144–153, May 2021, doi: 10.15294/sji.v8i1.26888. pp
y
j
[40]
I. F. Katili, F. D. Esabella, and A. Luthfiarta, “Pattern Recognition Of Javanese Letter Using
Template Matching Correlation Method,” J. Appl. Intell. Syst., vol. 3, no. 2, pp. 49–56, Dec. 2018,
doi: 10.33633/jais.v3i2.1954.
|
https://openalex.org/W4294557403
|
https://link.springer.com/content/pdf/10.1007/s10787-022-01033-8.pdf
|
English
| null |
A randomized controlled trial assessing the safety and efficacy of palmitoylethanolamide for treating diabetic-related peripheral neuropathic pain
|
Inflammopharmacology
| 2,022
|
cc-by
| 9,423
|
Inflammopharmacology (2022) 30:2063–2077
https://doi.org/10.1007/s10787-022-01033-8 Inflammopharmacology (2022) 30:2063–2077
https://doi.org/10.1007/s10787-022-01033-8 Inflammopharmacology ORIGINAL ARTICLE A randomized controlled trial assessing the safety and efficacy
of palmitoylethanolamide for treating diabetic‑related peripheral
neuropathic pain Emily Pickering1,2 · Elizabeth L. Steels1,2 · Kathryn J. Steadman1 · Amanda Rao3 · Lui Received: 28 June 2022 / Accepted: 30 June 2022 / Published online: 4 September 2022
© The Author(s) 2022 Abstract Background Peripheral neuropathy is a common complication of diabetes. The management of the associated neuropathic
pain remains difficult to treat.fi fi
Objective This study explored the safety, tolerability and efficacy of a palmitoylethanolamide (PEA) formulation in treating
diabetic-related peripheral neuropathic pain (PNP). Secondary outcomes included systemic inflammation, sleep and mood
changes in patients diagnosed with type 1 and type 2 diabetes and PNP. Design This study was a single-centre, quadruple-blinded, placebo-controlled trial with 70 participants receiving 600 mg of
PEA or placebo daily, for 8 weeks, with a 94% rate of study participation completion. Primary outcomes were neuropathic
pain and specific pain types (the BPI-DPN and NPSI). The secondary outcomes were sleep quality (MOS sleep scale), mood
(DASS-21), glucose metabolism and inflammation.i l
Results There was a significant reduction (P ≤ 0.001) in BPI-DPN total pain and pain interference, NPSI total score and
sub-scores, except for evoked pain (P = 0.09) in the PEA group compared with the placebo group. The MOS sleep problem
index and sub-scores significantly improved (P ≤ 0.001). DASS-21 depression scores significantly reduced (P = 0.03), but
not anxiety or stress scores. Interleukin-6 and elevated C-reactive protein levels significantly reduced in the PEA group
(P = 0.05), with no differences in fibrinogen between groups (P = 0.78) at treatment completion. There were no changes in
safety pathology parameters, and the treatment was well tolerated.l Conclusions The study demonstrated that the PEA formulation reduced diabetic peripheral neuropathic pain and inflamma-
tion along with improving mood and sleep. Further studies on the mechanistic effectiveness of PEA as an adjunct medicine
and as a monotherapy pain analgesic are warranted. Conclusions The study demonstrated that the PEA formulation reduced diabetic peripheral neuropathic pain and inflamma-
tion along with improving mood and sleep. Further studies on the mechanistic effectiveness of PEA as an adjunct medicine
and as a monotherapy pain analgesic are warranted. Clinical Trial Registration Registry name: Australian New Zealand Clinical Trials Registry (ANZCTR), Registration number:
ACTRN12620001302943, Registration link: https://anzctr.org.au/Trial/Registration/TrialReview.aspx?id=380826, Actual
study start date: 20 November 2020. Keywords Diabetic neuropathy · Pain · Neuropathic pain · Diabetes · Palmitoylethanolamide · PEA · Inflammation Keywords Diabetic neuropathy · Pain · Neuropathic pain · Diabetes · Palmitoylethanolamide · PEA · Inflammation 1
School of Pharmacy, University of Queensland, PACE
Precinct, 20 Cornwall Street, Wooloongabba, Brisbane,
QLD 4102, Australia * Elizabeth L. Steels
e.steels@uq.edu.au
Emily Pickering
e.pickering@uq.edu.au
Kathryn J. 1
School of Pharmacy, University of Queensland, PACE
Precinct, 20 Cornwall Street, Wooloongabba, Brisbane,
QLD 4102, Australia 1
School of Pharmacy, University of Queensland, PACE
Precinct, 20 Cornwall Street, Wooloongabba, Brisbane,
QLD 4102, Australia
2
Evidence Sciences Pty. Ltd., Brisbane, QLD, Australia
3
School of Human Movement and Nutrition Sciences,
University of Queensland, Brisbane, QLD 4102, Australia
4
Faculty of Medicine and Health, The University of Sydney,
Sydney, NSW, Australia Abstract Steadman
k.steadman@uq.edu.au
Amanda Rao
amanda@rdcglobal.com.au
Luis Vitetta
luis.vitetta@sydney.edu.au 2
Evidence Sciences Pty. Ltd., Brisbane, QLD, Australia 3
School of Human Movement and Nutrition Sciences,
University of Queensland, Brisbane, QLD 4102, Australia 4
Faculty of Medicine and Health, The University of Sydney,
Sydney, NSW, Australia (0123
1 3456789)
3 2064 E. Pickering et al. Abbreviations
AEs
Adverse events
ANOVA
Analysis of variance
ANZCTR
Australian New Zealand Clinical Trials
Registry
BMI
Body mass index
BPI-DPN
Brief pain inventory short form for diabetic
peripheral neuropathy
CRP
C-reactive protein
DASS-21
21-Item depression anxiety stress score
DPN
Diabetic peripheral neuropathy
DN4
Neuropathic pain diagnostic questionnaire
ENCB
Endocannabinoid
FBG
Fasting blood glucose
HbA1c
Glycosylated haemoglobin
HDPE
High-density polyethylene
IL-6
Interleukin-6
ITT
Intent to treat
MOS
Medical outcomes study—sleep scale
NAEs
N-Acylethanolamines
NF-KB
Nuclear factor kappa-light-chain-enhancer of
activated B cells
NHMRC
National Health and Medical Research
Council
NPSI
Neuropathic pain symptom inventory
PEA
Palmitoylethanolamide
PNP
Peripheral neuropathic pain
QOL
Quality of life
SD
Standard deviation
S-LANSS
Self-reported Leeds Assessment of Neuro-
pathic Symptoms and Signs
SPSS
Statistical package for the social sciences
TNF-α
Tumour necrosis factor alpha
VAS
Visual analogue scale These symptoms develop and worsen in the distal nerve
endings of the feet and in severe instances spread towards
the central parts of the body (Calcutt 2020). Small nerve
fibres make up 79–91% of peripheral nerve fibres (Said et al. 1992; Malik et al. 2005) and are more sensitive and prone
to damage than large nerve fibres. This results in the skin
of the toes and fingers to be the first areas affected by DPN
(Zochodne 2014). In DPN, continuously high serum glu-
cose levels damage small blood vessels, causing reduced
supply of oxygen and nutrients to the nerves (Bodman and
Varacallo 2021). This compromises the myelin sheath insu-
lating layer of the nerve fibres, particularly that of small
nerve fibres, progressing to loss of integrity (Malik 2014). The ongoing damage to small nerve fibres contributes to
the development of foot ulcerations (Gibbons et al. 2010),
reduced vasodilation due to pressure ulcers (Koïtka et al. 2004) and impaired heat and pain perception (Malik 2014). A recent murine model of diabetes (Yang et al. 2019) sug-
gested that variation to blood glucose levels weakens the
myelin sheath and nerve fibres and induces inflammation,
particularly increased pro-inflammatory cytokines such as
tumour necrosis factor alpha (TNF-α), interleukin-6 (IL-6)
and nuclear factor kappa-light-chain-enhancer of activated
B cells (NF-KB). Abstract Consequently, the progressive reduction in
the integrity of the myelin sheath has been demonstrated to
lead to increasing severity of DPN (Malik 2014). Abbreviations g
y
Maintaining control of blood glucose levels is a pri-
mary treatment strategy for DPN along with medications
for the management of neuropathic pain which includes
antiepileptics or anticonvulsants, tricyclic antidepressants,
serotonin–norepinephrine reuptake inhibitors and opioids
(Australian Medicines Handbook 2019). Reports demon-
strate that PEA has potential as an analgesic treatment for
DPN (D'amico et al. 2020). PEA belongs to the family of
N-acylethanolamines (NAEs) that are endogenous biologi-
cally active lipid mediators synthesized on demand by the
phospholipid membrane of cells (Alhouayek and Muccioli
2014; Mattace Raso et al. 2014). NAEs may assist in regu-
lating pain and inflammation with a neuroprotective effect
(Alhouayek and Muccioli 2014). PEA is also an endogenous
ligand of the endocannabinoid system (ENCB) with neuro-
modulator activity in the central nervous system (Clayton
et al. 2021). PEA has been reported to have an entourage
effect which can enhance the physiological effects of the
endogenous endocannabinoids such as N-arachidonoylethan-
olamine (i.e. anandamide). The overall effect is that through
N-arachidonoylethanolamine and the ENCB system, anti-
inflammatory and proapoptotic activities can inhibit pro-
inflammatory markers such as TNF-α and NF-KB (Sancho
et al. 2003). PEA is also considered to comprise a paral-
lel endocannabinoid signalling system without the adverse
effects such as those with exogenous endocannabinoids (e.g. THC). Introduction Diabetic peripheral neuropathy (DPN) is a neurodegenera-
tive disorder of the peripheral nervous system. It is the most
common complication of diabetes, estimated to affect up to
30% of this population, and is the leading cause of nerve
pain, disability due to foot ulceration and amputation, dis-
turbances to gait and fall-related injuries (Juster-Switlyk and
Smith 2016). DPN affects the motor, sensory and autonomic
nerves; however, it particularly targets the sensory nerves,
which are responsible for the transmission of information
including touch, temperature and injury pain (Bodman and
Varacallo 2021). The characteristics of the nerve pain range
from bouts of electric shocks and stabbing, abnormal sensa-
tions of tingling/pins and needles, spontaneous burning and
increased sensitivity to pressure and evoked pain by brush-
ing, pressure or cold temperatures (Bouhassira et al. 2004). 1 3 3 2065 A randomized controlled trial assessing the safety and efficacy of palmitoylethanolamide… Levagen+. The investigational product was formulated and
manufactured by Pharmako Biotechnologies Pty Ltd. under
Good Manufacturing Practice guidelines. The investigational
product was provided as clear capsules (size 00) contain-
ing 350 mg of Levagen+ providing no less than 300 mg
Palmidrol per capsule, which contains excipients polyglyc-
erol polyricinoleate (E476), coconut oil fractionated, lime
oil, olive oil, lecithin (sunflower and/or oat) (E322), silica
(E551), vitamin E), which were administered at a twice-daily
dose, equating to a total of dosing of 600 mg/day of PEA
over the 8-week study period. This dose has been shown to
be gut absorbed to an equivalent dose of 1.1025 g of micro-
nized PEA (Briskey et al. 2020). The matching placebo cap-
sule contained 350 mg of maltodextrin. The capsules were
packaged in high-density polyethylene (HDPE) bottles with
a HDPE lid. The packaging, labelling and dosage adminis-
tration of the placebo were the same as the investigational
product. Reduction in cellular PEA levels occurs due to prolonged
inflammation (Solorzano et al. 2009) and as a result from
nerve injury due to neuropathy (Franklin et al. 2003). An
early meta-analysis with a diverse variety of chronic neuro-
pathic pain conditions (three studies were specific for DPN)
demonstrated that PEA was progressively effective in reduc-
ing chronic neuropathic pain with the report concluding that
it has potential as a therapeutic strategy to manage chronic
neuropathic pain (Paladini et al. 2016). PEA, though, has been reported to be poorly bioavailable
with oral-gut administration, hindering pharmacological
efficacy (Gabrielsson et al. 2016). Introduction An increased absorbable
form has been developed (Briskey et al. 2020) that has been
reported to provide enhanced bioavailability and as such effi-
cacy and dosing. The aim of the current clinical study was
to determine whether an enhanced bioavailable formulation
of PEA was safe, tolerable and effective for managing DPN-
related pain and, moreover, and secondarily, this formulation
was effective in reducing inflammation and improving qual-
ity of life (QOL) associated with DPN, over an 8-week study
period in patients diagnosed with diabetes. Participant inclusion and exclusion criteria Participants meeting all inclusion and exclusion criteria
were invited to participate in the clinical study. Males or
females aged at least 18 years of age and diagnosed by a
treating physician with type 1 diabetes or type 2 diabetes and
with diabetic-related peripheral neuropathy were identified. Participants confirmed experiencing neuropathic pain by
scoring more than four on the Neuropathic Pain Diagnostic
Questionnaire (DN4) or scoring more than 12 on the Self-
reported Leeds Assessment of Neuropathic Symptoms and
Signs (S-LANSS). All participants were administering pre-
scribed anti-diabetic medications metformin and/or insulin. Some participants (31/66, 47%) were also co-administering
prescribed anti-diabetic medications with analgesic medica-
tions for pain. Participants were asked to continue taking
their prescribed medications for the duration of the study
and were permitted to take up to the maximum daily dose
(4 g/day) of paracetamol as pain rescue medication. Trial design The clinical study was an interventional, single-centre,
prospective, randomized, quadruple-blinded, placebo-con-
trolled, parallel study investigating the safety, tolerability
and efficacy of a PEA formulation on neuropathic pain. The secondary outcomes of improvement in inflammation
markers and sleep quality and improved quality of life when
administered as an adjunct analgesic to diabetic medications
were also measured over 8 weeks. The study included men
and women aged at least 18 years of age with a diagnosis of
type 1 or type 2 diabetes and who were prescribed glucose-
lowering medications including either or both metformin
and insulin. The study was conducted in Brisbane, Australia,
in accordance with the principles of The Declaration of Hel-
sinki and Australian Good Clinical Practice Guidelines. The
trial was registered with the Australian New Zealand Clinical
Trials Registry (ANZCTR) no.: ACTRN12620001302943. Potential participants were excluded if the peripheral neu-
ropathy was due to hereditary sensory neuropathy, vitamin
B12 or folate deficiency, paraneoplastic diseases, advanced
liver disease, kidney disease, hypothyroidism, prolonged
phenytoin, warfarin or immunosuppressive drug use. Par-
ticipants were also excluded if they were administering
herbal medicines for pain relief including, but not limited to,
turmeric/curcumin (Curcuma longa), boswellia (Boswellia
serrata), willow bark (Salix alba) or medicinal cannabis. Women who were pregnant, planning to become pregnant
or breastfeeding were excluded. Alcohol or substance abuse
health issues, allergy or sensitivity to any of the ingredi-
ents in the investigational product or any clinically relevant
abnormal findings, in the opinion of the investigators/clini-
cians, would make them not suitable for inclusion in the Investigational products The investigational product was a proprietary formulation
of palmitoylethanolamide (Palmidrol) which has previous
approval by the Therapeutic Goods Association as an active
ingredient in the listed medicines. The Palmidrol was pro-
vided by Gencor Pacific Ltd. and was combined with the
bioavailability enhancement technology system LipiSperse®
as defined by Briskey et al. (2022), under the brand name 1 3 2066 E. Pickering et al. study. All participants that met all criteria provided written
informed consent. (MOS). The MOS has been validated in patient populations
experiencing neuropathic pain (Hays et al. 2005; Rejas et al. 2007). The MOS sleep score includes a Sleep Problem Index
and the following dimensions of sleep disturbance: experi-
ence disturbance to sleep, achieving adequate sleep, sleep
quantity, daytime somnolence during the day, occurrence
of snoring and experiencing shortness of breath or headache
upon awakening. The 21-item Depression Anxiety Stress
Score (DASS-21) was also assessed at baseline and 8 weeks. This was a measure of mood distress along the three axes of
depression, anxiety and stress (Ng et al. 2007). study. All participants that met all criteria provided written
informed consent. Randomization, blinding and compliance Eligible participants were randomly allocated to one of two
study treatment groups with a ratio of 1:1. The study treat-
ments with PEA or placebo twice daily were provided to
participants in identical bottles labelled 001-070 sequen-
tially upon enrolment. Participants were independently
randomized prior to treatment medications being provided
to the clinic. The clinical trial was quadruple masked (i.e. participant, care provider, investigators, outcomes assessor)
to the allocation group of medications. Participant compli-
ance was assessed by the number of capsules taken, with
greater than 80% of the capsules consumed being accepted
as compliant with doses administered. Blood markers were assessed at baseline and at 8 weeks,
and these included HbA1c and fasting blood glucose (FBG),
as determined by the International Expert Committee guide-
lines (The Expert Committee on the Diagnosis and Classifi-
cation of Diabetes Mellitus, 2003, The International Expert
Committee 2009), and inflammation markers c-reactive
protein (CRP), IL-6 and fibrinogen with normal reference
ranges of 0–6 ml/L, 1.5–4.0 g/L and ≤ 1.8 pg/mL, respec-
tively. Blood safety markers were assessed at baseline and
8 weeks and included a full blood count, liver function tests,
platelets, electrolytes and kidney function. Product tolerabil-
ity was assessed regularly at each clinic interview and during
compliance checks by interviewers specifically documenting
changes to treatment medications, new symptom concerns,
new stressors or any mild, moderate or serious adverse
events that ensued. An adverse event form in the case file
forms was used to document adverse events. Adverse events
(AEs) were defined as any unfavourable changes in health,
including abnormal laboratory findings that occur in any
clinical trial participant, primarily during the clinical trial
period or within a specified period, following completion
of the clinical trial. The safety testing procedures followed
the NHMRC Safety Monitoring and Reporting in Clinical
Trials guidelines (National Health and Medical Research
Council 2016). Clinical study outcomes The primary outcome investigated the safety, tolerability and
effectiveness of PEA administered concurrently with pre-
scribed diabetic and pain medications to alleviate diabetic-
related neuropathic pain. The secondary outcomes included
a reduction in inflammatory markers and improvement in
mood and sleep quality. The primary outcome was considered as the overall
severity of neuropathic pain, assessed with the Brief Pain
Inventory Short Form for Diabetic Peripheral Neuropathy
(BPI-DPN). The BPI-DPN comprised a four-item Pain
Severity Score that rated worst pain, least pain, average pain
and present pain with a numerical rating score of 0 (i.e. no
pain) to 10 (i.e. worst imaginable pain). Pain was assessed
at baseline, 2, 4, 6 and 8 weeks. Additionally, the BPI-DPN
also contains a 10-item Pain Interference Score, which was
also measured at baseline, 4 and 8 weeks as a concomitant
primary outcome (Zelman et al. 2005). Additional pain asso-
ciated outcomes included the severity of the specific charac-
teristics of pain. The severity of the specific characteristics
of pain was assessed using the Neuropathic Pain Symptom
Inventory (NPSI) at baseline, 4 weeks and 8 weeks. The
NPSI includes 12 items that allow discrimination and quan-
tification of five distinct clinically relevant dimensions of
neuropathic pain syndromes, namely superficial spontaneous
burning, spontaneous pressing pain, paroxysmal pain (stab-
bing, pins and needles), evoked pain (mechanical brushing),
thermal allodynia (pressure)/hyperalgesia (contact with
cold) and dysesthesia/paraesthesia (pins and needles, tin-
gling). Each question provides a numerical rating score of 0
(i.e. no pain) to 10 (i.e. worst imaginable pain) (Bouhassira
et al. 2004). Correlations between glucose metabolism, pain,
sleep and mood There was a wide range in fasting blood glucose (FBG)
and glycosylated haemoglobin (HbA1c) levels, despite
the participants taking prescribed anti-diabetic medica-
tions (Table 3). The baseline FBG levels correlated with
HbA1c levels (r = 0.662, P < 0.001). There were correla-
tions between glucose metabolism and pain indices; par-
ticularly, a negative correlation between HbA1c and NPSI
evoked pain (r = − 0.251, P = 0.045) and FBG and BPI-
PN Pain Severity Index (r = − 0.322, P = 0.01), BPI-PN
total pain (r = − 0.370, P = 0.003), the NPSI paroxysmal
pain (r = − 0.266, P = 0.03), evoked pain (r = − 0.347,
P = 0.005) and paraesthesia/dysesthesia (r = − 0.257,
P = 0.04). Furthermore, CRP positively correlated with
both NPSI deep pain (r = 0.261, P = 0.04) and fibrinogen
levels (r = 0.387, P = 0.002). There was also a positive cor-
relation between BPI-PN pain interference and depression
scores (r = 0.431, P < 0.001), anxiety scores (r = 0.275,
P = 0.03), stress scores (r = 0.270, P = 0.03) and sleep
quality (r = 0.276, P = 0.03). Furthermore, CRP positively
correlated with both NPSI deep pain (r = 0.261, P = 0.04)
and fibrinogen levels (r = 0.387, P = 0.002) (Figs. 2 and 3). Statistical analysis An intent-to-treat (ITT) approach was used for data analy-
sis of the primary outcome, where all patients who were
randomized to receive treatment and completed baseline
and at least one subsequent assessment (including labora-
tory clinical markers) were included in the analysis. The
ITT single missing data points in those completing the study
were managed using the simple imputation method, with the
data from the last observation carried forward. Normality
between groups was tested using the Kolmogorov–Smirnov
and Shapiro–Wilk tests of normality. Demographics includ-
ing age, gender, weight, height and BMI were assessed for
statistical differences between treatment groups at baseline
and week 8 by Chi-square and presented with mean/SD. The pain scores were analysed for time and treatment effect
using repeated measures ANOVA. The QOL questionnaires
BPI-DPN, NPSI and MOS were analysed using t-tests (for
normally distributed data) and with Mann–Whitney U test
(for nonparametric data) between treatment groups. The
DASS-21 and the laboratory markers were assessed by
t-tests. Change scores were also used for data with wide
variations in values and/or significant differences at base-
line. Effect of confounders, including pain medications, was
assessed using linear regression analysis. The SPSS version
27 software was used for statistical analyses and for generat-
ing the graphical figures. Statistical significance was set at a
P < 0.05 (two-tailed). Sample size G*Power was used to calculate the sample size, based on
a statistical difference (P < 0.05) between two independent
means (active treatment and placebo groups, ratio 1:1) for
the primary outcome (BPI-DPN Pain Severity Index). With a
two-tailed, alpha error probability of 0.05 and a size effect of
0.8, the required total sample size was 29 participants per arm. Allowing for 15% rate of attrition, the required sample size for
recruitment was inflated to a total of 70 eligible participants
(in 1:1 ratio for active treatment group and placebo group (i.e. 35 per treatment group)). A previous effect size analysis of
PEA for treatment of pain using the differences between days
0 and 21 of PEA treatment for visual analogue scale (VAS)
scores from a meta-analysis (Clayton et al. 2021), with the
assumption that the VAS scores were normally distributed, Impact on sleep patterns was measured at baseline, 4 and
8 weeks using the Medical Outcomes Study—Sleep Scale 3 3 A randomized controlled trial assessing the safety and efficacy of palmitoylethanolamide… 2067 produced an estimated effect size (i.e. Cohen's d) value of 1.35
(95% CI 1.14–1.56) for the daily administration of 700 mg of
micronized PEA (Gabrielsson et al. 2016). diabetes (Table 1). The groups had a different ratio of men
to women, but there was no significant difference in base-
line health parameters of body weight, BMI, blood pres-
sure and lifestyle factors (Table 1). All participants were
taking prescribed diabetic medications, and the majority
were also taking prescribed pain medications as well as
other medications for co-morbidities (Table 1). Pain interference Table 3). By week 4, there was a significant difference between
the active treatment and placebo groups for total pain score
(P ≤ 0.001), superficial spontaneous pain (P ≤ 0.001), deep
pain (P = 0.03), paroxysmal pain (P ≤ 0.001) and paraesthesia
(P = 0.01). All these sub-scores were also found to be signifi-
cantly different at week 8 for total pain (P ≤ 0.001), superfi-
cial pain (P ≤ 0.001), deep pain (P = 0.002), paroxysmal pain
(P ≤ 0.001) and paraesthesia (P ≤ 0.001). The sub-score for
evoked pain, however, was not significantly different between
groups at week 4 (P = 0.25) yet was trending towards a signifi-
cant difference by week 8 (P = 0.09). Pain interference as measured by the BPI-DPN was not signifi-
cantly different between treatment groups at baseline (P = 0.15),
(Table 2). When evaluating the effect of treatment on pain
interference at week 4 and week 8, significant differences were
observed between the two treatment groups [F (1,64) = 12.60,
P ≤ 0.001] and the two time points [F (2,64) = 12.60, P ≤ 0.001]. At week 4, there was a significant difference between the means
of the active treatment and placebo group (P = 0.001). Week
8 means also were significantly different between groups
(P ≤ 0.001) with a greater change from baseline in the active
group (−1.90) than the placebo group (−0.39). Pain severity A total of 235 prospective participants were recruited
through public social media activities. Prospective par-
ticipants registered their interest initially through an online
screening form, followed by an e-consult of a comprehen-
sive assessment including medical history and medica-
tions to establish eligibility. A total of 70 participants aged
between 32 and 75 years met the inclusion and exclusion
criteria and, following the provision of written informed
consent, were enrolled in the study. Pain severity as measured by the Brief Pain Inventory
Short Form for Diabetic Peripheral Neuropathy (BPI-
DPN) was not statistically different between groups
at baseline, (P = 0.46) with most participants in both
groups exhibiting mild to moderate pain (score of 2–7),
(Table 2). Repeated measures ANOVA revealed a both
time [F (4,64) = 21.03, P < 0.001] and treatment [F
(1,64) = 23.52, P < 0.001] effect for the investigational
product over 8 weeks of the study. Further analysis indi-
cated that there was a significant difference in pain severity
between groups at week 2 (P = 0.002), which continued
at weeks 4, 6 and 8 (P < 0.001) (Table 2). Interestingly,
a progressive and significant improvement in treatment
group over placebo group in pain scores was observed as
the study progressed. The final analysis group included 66 participants
(Fig. 1, Table 1), 33 in each arm. Four participants were
excluded from analysis: three due to incomplete baseline
assessments and one due to pre-existing (unknown) dis-
ease that was an exclusion criterion (Fig. 1). The cohort
included three participants with type 1 and 62 with type 2 1 3 2068 E. Pickering et al. Fig. 1 Participant progress CONSORT flow chart Fig. 1 Participant progress CONSORT flow chart Effect of treatment on neuropathic pain symptoms
(NPSI) The use of pain medication was not associated with any
difference in BPI-PN pain severity, NPSI total and sub-
scores at completion of treatment in either group. The
mean pain medication index expressed as the number of All measurements of neuropathic pain symptoms as measured
by the NPSI were similar between groups at baseline (P ≥ 0.05, 1 3 A randomized controlled trial assessing the safety and efficacy of palmitoylethanolamide… A randomized controlled trial assessing the safety and efficacy of palmitoylethanolamide… 2069 1 Pearson Chi-Square test; all other P values assessed by two-tailed t test; statistical significance P < 0.05
2 Additional diabetic medications: semaglutide, empagliflozin, linagliptin, dulaglutide, dapagliflozin, acar-
bose, gliclazide MR, vildagliptin, sitagliptin, exenatide
3 Other pain medications: meloxicam (1) and tramadol (1), oxycodone (2)
4 May have been prescribed for pain relief and/or mood support
Active group
n = 35
Placebo group
n = 35
P value1
Total (n) %
33 (50%)
33 (50%)
1.000
Men (n) %
20 (60%)
15 (40%)
0.221
Women (n) %
13 (42%)
18 (58%)
Total Age (Av, range)
65.5 [53–79]
61.5 [32–75]
0.056
Men Age (Av, range)
67.8 [53–79]
61.2 [32–75]
0.046
Women Age (Av, range)
62.1 [53–74]
61.8 [50–71]
0.92
Total BMI (Av, range)
31.2 [23.5–42.3]
31.5 [22.9–39.5]
0.80
Men BMI (Av, range)
29.9 [23.5–37.4]
31.4 [25.1–37.8]
0.27
Women BMI (Av, range)
33.3 [24.4–42.3]
31.7 [22.9–39.5]
0.39
Medications taken at baseline
n = 33
n = 33
Metformin only
23
21
0.669
Insulin only
6
8
0.562
Metformin and Insulin
4
4
1.000
Additional diabetic medications2
19
22
0.626
Analgesic medications
Ibuprofen
2
4
0.392
Paracetamol
7
7
1.000
Aspirin
1
3
0.302
Pregabalin
7
5
0.523
Other pain medications3
2
2
1.000
Tricyclic antidepressants4
4
4
1.000
Blood pressure medications
17
19
0.655
Cholesterol medication
16
14
0.621
Anti-depressant/anti-anxiety5
7
13
0.370 1 Pearson Chi-Square test; all other P values assessed by two-tailed t test; statistical significance P < 0.05 2 Additional diabetic medications: semaglutide, empagliflozin, linagliptin, dulaglutide, dapagliflozin, acar
bose, gliclazide MR, vildagliptin, sitagliptin, exenatide non-treatment medication found no difference between
treatment groups for total medications (prescribed and
rescue) or rescue medications used. analysis of participants with CRP levels of ≥ 5.0 mg/L at
baseline was similar at baseline (P = 0.18); however, at
8 weeks the active treatment and placebo group demon-
strated a significant difference between means (P = 0.05). Effect of treatment on neuropathic pain symptoms
(NPSI) For IL-6, the groups were also similar at baseline
(P = 0.44); however, a significant difference was recorded
at 8 weeks between the active treatment group and the
placebo group (P = 0.04). Effect of treatment on glucose metabolism
and inflammatory markers The treatment groups varied widely in glucose metabo-
lism parameters (FBG and HbA1c) and the inflammatory
markers (IL-6, fibrinogen and CRP) (Table 4). All mark-
ers were not significantly different at baseline (P > 0.05). At 8 weeks, there was no significant difference between
the active treatment and placebo group for fasting blood
glucose, HbA1c, fibrinogen and CRP when analysed as
a whole group across all levels (P > 0.05). A sub-group Effect of treatment on sleep The groups were similar at baseline for all the Medical
Outcomes Study—Sleep Scale (MOS) subscale scores
(P ≥ 0.05), (Table 5). At 8 weeks, there was a signifi-
cant difference between the active treatment group com-
pared with the placebo group in the sub-scores of sleep 1 3 2070 E. Pickering et al. sturbance (P = 0.001), sleep adequacy (P = 0.001),
ytime somnolence (P ≤ 0.001), shortness of breath or
adache (P = 0.04) and the total sleep problem index
≤ 0.001). There was no significant change after 8 weeks
Effect of treatment on the mood parameters
depression, anxiety and stress
Groups were statistically different at baseline for de
2 BPI-PN (A) Pain Severity scores at baseline, 2, 4, 6 and 8 weeks and (B) Pain interference score at baseline, 4 and 8 weeks scores f
ve treatment group and the placebo group Fig. 2 BPI-PN (A) Pain Severity scores at baseline, 2, 4, 6 and 8 weeks and (B) Pain interference score at baseline, 4 and 8 weeks scores for the
active treatment group and the placebo group Effect of treatment on the mood parameters
depression, anxiety and stress disturbance (P = 0.001), sleep adequacy (P = 0.001),
daytime somnolence (P ≤ 0.001), shortness of breath or
headache (P = 0.04) and the total sleep problem index
(P ≤ 0.001). There was no significant change after 8 weeks
for sleep quantity (P = 0.52) or snoring (P = 0.22). Groups were statistically different at baseline for depres-
sion (active: 2.06 ± 2.95, placebo: 3.97 ± 3.75, P = 0.02) and 1 3 3 A randomized controlled trial assessing the safety and efficacy of palmitoylethanolamide… 2071 Fig. 3 Neuropathic Pain Symptom Inventory (NPSI) total and sub-scores for the active treatment group and the placebo group at 8 weeks Fig. 3 Neuropathic Pain Symptom Inventory (NPSI) total and sub-scores for the active treatment group and the placebo group at 8 weeks Pain Symptom Inventory (NPSI) total and sub-scores for the active treatment group and the placebo group at 8 weeks Fig. 3 Neuropathic Pain Symptom Inventory (NPSI) total and sub-scores for the active treatment group and the pl anxiety scores (active: 0.64 ± 1.34, placebo: 2.03 ± 2.89,
P = 0.02), but not stress scores (active: 2.94 ± 3.46, pla-
cebo: 4.15 ± 3.41, P = 0.16), so analysis was performed on
change scores. At 8 weeks, change scores were significantly
different between the active treatment and placebo groups
for depression (active: −1.27 ± 4.41 vs placebo: 0.09 ± 1.74,
P = 0.02, d = −0.562, CI = −1.052 to −0.067) and were
trending towards significance for stress (active: −1.36 ± 2.46
vs −0.39 ± 2.05, P = 0.09). There was no significant change anxiety scores (active: 0.64 ± 1.34, placebo: 2.03 ± 2.89,
P = 0.02), but not stress scores (active: 2.94 ± 3.46, pla-
cebo: 4.15 ± 3.41, P = 0.16), so analysis was performed on
change scores. At 8 weeks, change scores were significantly
different between the active treatment and placebo groups
for depression (active: −1.27 ± 4.41 vs placebo: 0.09 ± 1.74,
P = 0.02, d = −0.562, CI = −1.052 to −0.067) and were
trending towards significance for stress (active: −1.36 ± 2.46
vs −0.39 ± 2.05, P = 0.09). There was no significant change in anxiety levels for either group by the end of 8 weeks
(active: −0.333 ± 0.85 vs placebo: −0.545 ± 1.23, P = 0.42). Safety and tolerability of treatment There were no changes observed for the independently
reported laboratory markers for participant electrolytes, kid-
ney and liver function. It is noteworthy, though, that blood
glucose levels in both groups rose significantly over 8 weeks 1 3 E. Pickering et al. 2072 Table 2 BPI-PN pain outcome scores for active treatment group and placebo group at baseline, 2, 4, 6 and 8 weeks
1 Score given as average ± standard deviation
2 Shapiro–Wilk distribution test found these data (in one or both arms) to be not normally distributed; tests of significance were performed non-
parametrically with Mann–Whitney U. Active n = 33, Placebo n = 33. Statistical significance set at P ≤ 0.05
Intention to treat
Outcome
Time Point
Group
Mean ± SD1
Range
P value
Change from
baseline
Effect size [95% CI]
BPI Severity score2
Baseline
Active
4.77 ± 1.45
2.8–8.8
0.46
−0.135 [−0.617 to 0.349]
Placebo
4.96 ± 1.37
2.3–8.0
Week 2
Active
3.03 ± 1.39
1.0–7.9
0.002
−1.75
−0.790 [−1.289 to −0.286]
Placebo
4.14 ± 1.48
1.3–8.3
−0.80
Week 4
Active
2.49 ± 1.50
0.8–6.8
<0.001
−2.28
−1.131 [−1.648 to −0.607]
Placebo
4.27 ± 1.64
1.8–8.0
−0.70
Week 6
Active
2.30 ± 1.84
0.0–7.8
<0.001
−2.47
−1.324 [−1.853 to −0.785]
Placebo
4.54 ± 1.53
2.0–8.0
−0.42
Week 8
Active
1.72 ± 1.51
0.3–7.3
<0.001
−3.06
−1.811 [−2.381 to −1.231]
Placebo
4.19 ± 1.20
1.8–7.0
−0.77
BPI
Interference score2
Baseline
Active
3.49 ± 1.90
0.6–7.9
0.15
−0.398 [−0.884 to 0.091]
Placebo
4.26 ± 1.97
0.9–7.9
Week 4
Active
2.45 ± 1.85
0.1–6.4
0.001
−1.04
−0.882 [−1.385 to −0.373]
Placebo
4.11 ± 1.90
0.7–8.0
−0.15
Week 8
Active
1.60 ± 1.66
0.0–5.9
<0.001
−1.90
−1.303 [−1.831 to −0.766]
Placebo
3.87 ± 1.83
0.9–6.7
−0.39 e 2 BPI-PN pain outcome scores for active treatment group and placebo group at baseline, 2, 4, 6 and 8 weeks 1 Score given as average ± standard deviation g
g
2 Shapiro–Wilk distribution test found these data (in one or both arms) to be not normally distributed; tests of significance were performed non-
parametrically with Mann–Whitney U. Active n = 33, Placebo n = 33. Statistical significance set at P ≤ 0.05 reduction in spontaneous burning pain, spontaneous pres-
sure pain, paroxysmal pain and paraesthesia with no relief
observed from evoked pain, which is the response to touch,
pressure and cold. Safety and tolerability of treatment Furthermore, the reductions in neurologi-
cal pain were associated with lower levels of the inflamma-
tion markers IL-6 and CRP as well as a lower pain interfer-
ence with life activities (i.e. walking, gardening, cooking,
household duties), reduction in depression symptoms and
improvement in quality of sleep. of the clinical trial period (change in fasting glucose from
baseline: active + 0.88 mmol/L (+11.12%) P = 0.129, pla-
cebo + 0.95 mmol/L (+10.85%) P = 0.041, Table 4). Haema-
tology parameters remained constant throughout the 8-week
clinical trial except for a significant difference in the eosino-
phil count (0.04 vs −0.02, P = 0.01). There were several mild
and short-duration adverse events reported by participants
during the 8-week study period, which resolved over a few
days without withdrawing from the study. These included
intermittent mild headaches (n = 1, active treatment; n = 1,
placebo), episodes of constipation (n = 1, active treatment),
urticaria for 5 days (n = 1, active treatment), a severe fatigue
episode (n = 1, placebo group) and respiratory infections not
requiring additional medications (n = 3, placebo group) dur-
ing the study. All adverse events resolved during the 8-week
study period, and no participant was withdrawn from the
study because of an adverse event. The results of this study build on data from two previ-
ous clinical trials in patients diagnosed with DPN, albeit,
with different doses of micronized PEA. It is notable that
the analgesic effect of the PEA was significant from as
early as 2 weeks. In a study that administered 600 mg of a
micronized (non-emulsified) PEA, a significant reduction in
neuropathic pain associated with DPN was observed after
30 days and 60 days (Schifilliti et al. 2014). Furthermore,
the use of a higher dose of 1200 mg/day of the micronized
(non-emulsified) PEA resulted in significantly reduced dia-
betic and traumatic chronic neuropathic pain after 40 days
of treatment (Cocito et al. 2014). The results of the present
study highlight the variability that can be encountered with
different pain manifestations such as occurs with spontane-
ous, evoked, paroxysmal pain between individuals diagnosed
with diabetes, which may reflect different mechanisms of
the disease (Attal et al. 2008). In the current and previous
studies, PEA was observed to have a significant effect in Discussion The results of this study demonstrated that the formula-
tion of PEA was safe, tolerable and demonstrated analgesic
efficacy for mild to moderate neuropathic pain in patients
diagnosed with diabetic-related peripheral neuropathy when
tested over an 8-week period over placebo. The treatment
reduced pain in different sensory nerves as shown by the 1 3 3 A randomized controlled trial assessing the safety and efficacy of palmitoylethanolamide… 2073 Table 3 NPSI Pain outcome scores for active treatment group and placebo group at baseline, 4 and 8 weeks
Active n = 33, Placebo n = 33. Statistical significance set at P = < 0.05
1 Shapiro–Wilk distribution test found these data (in one or both arms) to be not normally distributed
Tests of significance are performed nonparametrically with Mann–Whitney U scores given as average ± standard deviation
Outcome
Group
Baseline
P value
4 weeks
P value
Change at
4 weeks
Effect size [95% CI]
8 weeks
P value
Change at
8 weeks
Effect size [95% CI]
Total pain score
Active
0.48 ± 0.17
0.86
0.29 ± 0.12
<0.001
−0.18
−1.061 [−1.574 to −0.541]
0.22 ± 0.13
<0.001
−0.25
−1.587 [−2.137 to −1.027]
Placebo
0.47 ± 0.17
0.43 ± 0.13
−0.04
0.43 ± 0.13
−0.04
Superficial pain
Active
5.94 ± 2.37
0.11
4.24 ± 2.15
<0.001
−1.70
−1.059 [−1.571 to −0.539]
3.21 ± 2.07
<0.001
−2.72
−1.651 [−2.207 to −1.086]
Placebo
6.85 ± 2.18
6.21 ± 1.52
−0.63
6.24 ± 1.56
−0.60
Deep pain
Active
5.79 ± 4.44
0.881
3.68 ± 3.33
0.03
−2.10
−0.562 [−1.052 to −0.067]
2.79 ± 2.86
0.0021
−3.00
−0.841 [−1.342 to −0.334]
Placebo
6.06 ± 4.45
5.79 ± 4.13
−0.27
5.55 ± 3.66
−0.51
Paroxysmal pain
Active
8.65 ± 4.30
0.911
4.29 ± 3.05
<0.001
−4.36
−0.991 [−1.500 to −0.476]
2.79 ± 2.35
<0.001
−5.86
−1.528 [−2.073 to −0.973]
Placebo
8.56 ± 4.34
7.55 ± 3.51
−1.02
7.22 ± 3.37
−1.33
Evoked pain
Active
8.99 ± 6.11
0.551
7.02 ± 5.28
0.25
−1.97
−0.283 [−0.768 to 0.203]
6.27 ± 4.82
0.09
−2.72
−0.423 [−0.909 to 0.067]
Placebo
8.25 ± 5.79
8.51 ± 5.19
0.25
8.39 ± 5.20
0.14
Paraesthesia
Active
8.96 ± 3.02
0.201
4.77 ± 3.00
0.01
−4.18
−0.642 [−1.135 to −0.145]
3.12 ± 2.57
<0.001
−5.83
−1.473 [−2.014 to −0.922]
Placebo
7.91 ± 3.47
6.77 ± 3.22
−1.14
7.08 ± 2.79
−0.83 1 3 2074 E. Pickering et al. Discussion Table 4 Effect of treatment on glucose metabolism and inflammatory markers at for the active treatment group and placebo group at baseline
and week 8
Active treatment group n = 33, Placebo group n = 33. Statistical significance set at P ≤ 0.05
1 Shapiro–Wilk distribution test found these data (in one or both arms) to be not normally distributed; tests of significance were performed non-
parametrically with Mann–Whitney U
Test (Reference Range)
Group
Baseline
Mean ± SD
Range
P value
8 Weeks
Mean ± SD
Range
P value
Fasting Blood Glucose
(3.0–7.7 mmol/L)
Active
7.91 ± 2.28
3.8–12.8
0.23
8.89 ± 3.17
5.1–20.2
0.791
Placebo
8.75 ± 3.26
3.3–19.0
9.13 ± 4.23
4.4–25.0
HbA1c mmol/L
(48–53 mmol/mol)
Active
58.35 ± 15.23
39.0–95.0
0.42
58.32 ± 16.47
37.0–96.0
0.531
Placebo
62.06 ± 20.73
36.0–121.0
62.97 ± 23.62
35–151
HbA1c %
(6.5–7%)
Active
7.49 ± 1.39
5.7–10.8
0.42
7.48 ± 1.51
5.5–11.0
0.531
Placebo
7.83 ± 1.90
5.4–13.3
7.91 ± 2.15
5.4–15.9
C-reactive Protein
(0–6 mg/L)
Active
5.32 ± 3.25
2.5–12.0
0.911
4.63 ± 3.19
2.5–12.0
0.261
Placebo
5.97 ± 4.91
2.5–21.0
6.13 ± 4.74
2.5–18.0
C-reactive protein
> 5.0 mg/L
Active
8.38 ± 1.89
5.0–12.0
0.18
6.17 ± 3.35
2.5–12.0
0.05
Placebo
10.13 ± 4.60
5.0–21.0
9.43 ± 5.07
2.5–18.0
Interleukin-6
(< 6 pg/mL)
Active
4.93 ± 1.79
1.0–9.0
0.44
4.13 ± 2.32
1.0–12.0
0.041
Placebo
4.52 ± 2.27
1.0–12.0
5.44 ± 3.02
1.0–18.0
Fibrinogen
(1.50–4.00 g/L) 1
Active
3.73 ± 0.83
2.31–6.46
0.531
3.55 ± 0.75
1.85–5.08
0.78
Placebo
3.56 ± 0.85
2.15–4.93
3.61 ± 0.96
1.17–5.98 glucose metabolism and inflammatory markers at for the active treatment group and placebo group at baseline Active treatment group n = 33, Placebo group n = 33. Statistical significance set at P ≤ 0.05
1 Shapiro–Wilk distribution test found these data (in one or both arms) to be not normally distributed; tests of significance were performed non-
parametrically with Mann–Whitney U levels of blood glucose levels also weaken the myelin
sheath and nerve fibres (Magrinelli et al. 2015). This pro-
cess, alongside the production of inflammatory markers, is
thought to progress the development of peripheral neurop-
athy (Jin and Park 2018). The inflammatory markers IL-6
and CRP were significantly reduced after treatment with
PEA in this study, and it is suggested that reducing inflam-
mation was part of the mechanism underlying the observed
reduction in neuropathic pain. Discussion This is supported by recent
cellular macrophage studies that have reported that PEA
inhibits both TNF-α and IL-6 release (Del Re et al. 2021). It is noteworthy that there was a correlation in this study
between elevated baseline fasting blood glucose and pain,
also HbA1c and pain as well as CRP and pain. These data
draw a link between effective management of blood glu-
cose levels in pre-diabetics and diabetics with possible
prevention and reduction in peripheral neuropathic pain. reducing the specific pain characteristic of paraesthesia/dys-
esthesia. PEA has also previously been shown to be effec-
tive in ameliorating pain associated with nerve compression
syndromes such as sciatic pain (Keppel Hesselink and Kop-
sky 2015; Domínguez et al. 2012), carpal tunnel syndrome
(Conigliaro et al. 2011) and Charcot–Marie–Tooth neu-
ropathy (Putzu 2016) as well as knee osteoarthritis (Steels
et al. 2019). The existing in-vitro, animal and human clinical
studies demonstrate and support the efficacy of PEA in the
treatment of algesia in neuropathies resulting from various
causes (Paladini et al. 2016). There is a strong correlation between pain and depression
(Alghafri et al. 2020; Vas and Papanas 2020). The current
study further confirms that PEA reduced depression symp-
toms when administered as an adjunct to citalopram (Ghaz-
izadeh-Hashemi et al. 2018). Furthermore, we also report
that the administration of PEA was associated with improve-
ments in sleep adequacy, reduction in night-time sleep dis-
turbances and daytime somnolence, confirming reports of
improved overall sleep quality in surgical patients (Evange-
lista et al. 2018) and those patients with poor sleep quality,
albeit without pain (Putzu 2016). A proportion of participants in this study were taking
a range of prescribed anti-diabetic medications and medi-
cations to manage the neuropathic pain, which showed
that PEA being effective as standalone treatment and
an adjunct medication for pain relief. It was noteworthy
that there was a significant placebo response observed,
although PEA was significantly different to placebo, a phe-
nomenon often seen in pain studies. Further research into
effectiveness of PEA as an adjunct to long-term pain medi-
cation should be further investigated. There are limitations
in this study that must be considered. Discussion The cohort was pri-
marily type 2 diabetic and further research is needed with Prolonged hyperglycaemia observed in early or poorly
controlled diabetes presents a metabolic disease with
inflammatory sequelae and with upregulation of nuclear
factor-KB (NF-KB), which subsequently increases TNF-α
and drives a follow-on production of IL-6 and CRP (Yang
et al. 2019; Mu et al. 2017, Patel and Santani 2009). Recent animal studies suggest that varying and elevated 1 3 3 A randomized controlled trial assessing the safety and efficacy of palmitoylethanolamide… 2075 1
Table 5 MOS sleep sub-scores for active treatment group and placebo group at baseline, 4 weeks and 8 weeks
MOS score ratings depict the level of impact of each subcategory. Questions are a mix of positively (*) and negatively phrased (**) questions
1 Shapiro–Wilk distribution test found these data (in one or both arms) to be not normally distributed; tests of significance are performed nonparametrically with Mann–Whitney U. Active treat-
ment group n = 33, Placebo group n = 33. Conclusion Attal N, Fermanian C, Fermanian J, Lanteri-Minet M, Alchaar H, Bou-
hassira D (2008) Neuropathic pain: are there distinct subtypes
depending on the aetiology or anatomical lesion? Pain 138:343–
353. https://doi.org/10.1016/j.pain.2008.01.006 An 8-week randomized clinical study concluded that the
PEA formulation investigated reduced diabetic-related neu-
ropathic pain and was associated with a reduction in inflam-
mation and depression. In addition, there was subsequent
improvement in sleep quality. PEA was also well tolerated
and reported to be safe as an adjunct in patients prescribed
metformin and or insulin for the management of either type
1 or type 2 diabetes. Australian Medicines Handbook (2019) Neuropathic pain. Australian
Medicines Handbook. Australian Medicines Handbook Pty Ltd,
Adelaide Bodman MA, Varacallo M (2021) Peripheral diabetic neuropathy
[Online]. Treasure Island, Florida: StatPearls Publishing. https://
www.ncbi.nlm.nih.gov/books/NBK442009/. Accessed 20 Sep 2021 Bouhassira D, Attal N, Fermanian J, Alchaar H, Gautron M, Masquelier
E, Rostaing S, Lanteri-Minet M, Collin E, Grisart J, Boureau F
(2004) Development and validation of the neuropathic pain symp-
tom inventory. Pain 108:248–257. https://doi.org/10.1016/j.pain.
2003.12.024 Author contributions Roles of the authors of this manuscript were as
follows: ES and LV were involved in the study design; ES and EP col-
lected the data; ES, EP and AR analysed the data; EP, ES, AR, KS and
LV prepared the manuscript. All authors approved the submission of
the final version of the manuscript. Briskey D, Mallard A, Rao A (2020) Increased absorption of palmi-
toylethanolamide using a novel dispersion technology system
(LipiSperse®). J. Nutraceuticals Food Sci 5:1–6. https://doi.org/
10.36648/nutraceuticals.5.2.3 Calcutt N (2020) Diabetic neuropathy and neuropathic pain: a (con)
fusion of pathogenic mechanisms? Pain 161:S65-s86. https://doi.
org/10.1097/j.pain.0000000000001922 Funding Open Access funding enabled and organized by CAUL and
its Member Institutions. Gencor Pacific Ltd., Hong Kong, was the trial
sponsor. The funding source had no role in the design of the clini-
cal trial, albeit the sponsor monitored its implementation. Further, the
sponsor had no influence over the analysis, reporting and interpretation
of the data. EP was supported by the University of Queensland Gradu-
ate School scholarship. Clayton P, Hill M, Bogoda N, Subah S, Venkatesh R (2021) Palmitoy-
lethanolamide: a natural compound for health management. Int J
Mol Sci 22:5305. https://doi.org/10.3390/ikms22105305 Cocito D, Peci E, Ciaramitaro P, Merola A, Lopiano L (2014) Short-
term efficacy of ultramicronized palmitoylethanolamide in periph-
eral neuropathic pain. Pain Res Treat 2014:854560–854560. Discussion permitted by statutory regulation or exceeds the permitted use, you will
need to obtain permission directly from the copyright holder. To view a
copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. Discussion Statistical significance set at P ≤ 0.05
MOS
sub-scale
Group
Baseline
Mean ± SD
(range)
P value 4 Weeks
Mean ± SD
(range)
P value Change at
4 weeks
Effect size
[95% CI]
8 Weeks
Mean ± SD
(range)
P value
Change at
8 weeks
Effect size
[95% CI]
Sleep distur-
bance*
Active
3.54 ± 0.79
(1.75–4.75)
0.64
3.92 ± 0.75
(2.00–5.00)
0.071
0.37
0.458
[−0.033 to 0.945]
4.20 ± 0.56
(2.25–5.00)
0.0011
0.65
0.843
[0.336 to 1.344]
Placebo 3.64 ± 0.91
(1.75–5.00)
3.55 ± 0.84
(2.00–5.00)
−0.09
3.62 ± 0.79
(2.00–4.75)
−0.02
Sleep adequacy*
Active
3.50 ± 1.47
(1.50–6.00)
0.391
2.59 ± 1.27
(1.00–5.50)
0.04
−0.91
−0.507
[−0.995 to −0.014]
2.12 ± 0.91
(1.00–4.00)
0.0011
−1.38
−0.831
[−1.331 to 0.324]
Placebo 3.15 ± 1.29
(1.00–6.00)
3.24 ± 1.31
(1.00–5.50)
0.09
3.05 ± 1.28
(1.50–6.00)
−0.11
Sleep quantity*
Active
6.02 ± 1.58
(2.00–1.00)
0.09
6.74 ± 1.43
(3.00–1.00)
0.69
0.72
6.83 ± 1.28
(4.00–9.00)
0.52
0.82
Placebo 6.67 ± 1.53
(4.00–1.00)
6.88 ± 1.38
(4.00–1.00)
0.21
7.04 ± 1.39
(4.00–1.00)
0.38
Daytime somno-
lence*
Active
4.44 ± 1.14
(2.33–6.00)
0.111
5.05 ± 0.89
(3.00–6.00)
0.001
0.61
0.923
[0.411 to 1.428]
5.30 ± 0.81
(3.33–6.00)
<0.0011
0.86
1.178
[0.651 to 1.698]
Placebo 3.90 ± 1.34
(1.00–6.00)
3.99 ± 1.36
(1.00–6.00)
0.09
4.08 ± 1.23
(1.00–6.00)
0.18
Snoring**
Active
3.67 ± 1.90
(1.00–6.00)
0.071
3.94 ± 1.85
(1.00–6.00)
0.43
0.27
3.91 ± 1.76
(1.00–6.00)
0.221
0.24
Placebo 4.48 ± 1.66
(1.00–6.00)
4.39 ± 1.48
(1.00–6.00)
−0.09
4.45 ± 1.42
(2.00–6.00)
−0.03
Shortness of
breath or head-
ache**
Active
5.76 ± 0.50
(4.00–6.00)
0.421
5.82 ± 0.46
(4.00–6.00)
0.11
0.06
5.93 ± 0.24
(5.00–6.00)
0.041
0.18
0.540
[0.047 to 1.029
Placebo 5.36 ± 1.32
(1.00–6.00)
5.36 ± 1.17
(2.00–6.00)
0.00
5.54 ± 1.00)
(2.00–6.00)
0.18
Sleep Problem
Index**
Active
4.01 ± 0.35
(3.22–4.78)
0.25
4.12 ± 0.37
(3.11–5.00)
0.01
0.11
4.20 ± 0.29
(3.11–4.78)
<0.001
0.18
0.859
[0.351 to 1.361]
Placebo 3.82 ± 0.58
(2.11–4.67)
3.82 ± 0.53
(2.56–4.56)
0.0003
3.86 ± 0.46
(2.78–4.44)
0.04 2076 E. Pickering et al. larger cohorts of both type 1 and type 2 diabetics. The
cohort had varying levels diabetic control (as measured by
HbA1c); therefor, larger studies may determine whether
glycaemic levels impacted on the effectiveness of PEA. In addition, specific assessment of individual motor, sen-
sory and autonomic nerve fibres is required to understand
the full clinical potential of PEA in diabetic peripheral
neuropathy. References Alghafri R, Gatt A, Formosa C (2020) Depression symptoms in
patients with diabetic peripheral neuropathy. Rev Diabet Stud
16:35–40. https://doi.org/10.1900/rds.2020.16.35l Alhouayek M, Muccioli G (2014) Harnessing the anti-inflammatory
potential of palmitoylethanolamide. Drug Discov Today 19:1632–
1639. https://doi.org/10.1016/j.drudis.2014.06.007 Conclusion https://doi.org/10.1155/2014/854560 Conigliaro R, Drago V, Foster P, Schievano C, Di Marzo V (2011) Use
of palmitoylethanolamide in the entrapment neuropathy of the
median in the wrist. Minerva Med 102:141–147 Data Availability Data described in the manuscript, code book and ana-
lytical code will be made available pending application and approval. D’amico R, Impellizzeri D, Cuzzocrea S, Di Paola R (2020)
ALIAmides update: palmitoylethanolamide and its formulations
on management of peripheral neuropathic pain. Int J Mol Sci
21:5330. https://doi.org/10.3390/ijms21155330 Declarations Conflict of interest None of the authors had any conflict of interest in
relation to this manuscript. Domínguez C, Martín A, Ferrer F, Puertas M, Muro A, González J, Pri-
eto J, Taberna I (2012) N-palmitoylethanolamide in the treatment
of neuropathic pain associated with lumbosciatica. Pain Manag
2:119–124. https://doi.org/10.2217/pmt.12.5 Ethics approval This study was approved by the National Institute of
Integrative Medicine Human Research Ethics Committee, trial refer-
ence number 0076E_2020. Evangelista M, Cilli V, De Vitis R, Militerno A, Fanfani F (2018)
Ultra-micronized palmitoylethanolamide effects on sleep-wake
rhythm and neuropathic pain phenotypes in patients with carpal
tunnel syndrome: an open-label, randomized controlled study. CNS Neurol Disord Drug Targets 17:291–298. https://doi.org/
10.2174/1871527317666180420143830 Open Access This article is licensed under a Creative Commons Attri-
bution 4.0 International License, which permits use, sharing, adapta-
tion, distribution and reproduction in any medium or format, as long
as you give appropriate credit to the original author(s) and the source,
provide a link to the Creative Commons licence, and indicate if changes
were made. The images or other third party material in this article are
included in the article's Creative Commons licence, unless indicated
otherwise in a credit line to the material. If material is not included in
the article's Creative Commons licence and your intended use is not Franklin A, Parmentier-Batteur S, Walter L, Greenberg DA, Stella
N (2003) Palmitoylethanolamide increases after focal cerebral
ischemia and potentiates microglial cell motility. J Neurosci
23:7767–7775. https://doi.org/10.1523/JNEUROSCI.23-21-
07767.2003 1 3 3 A randomized controlled trial assessing the safety and efficacy of palmitoylethanolamide… 2077 Putzu G (2016) Efficacy of ultramicronized palmitoylethanolamide on
the clinical symptoms of Charcot-Marie-tooth neuropathy. Archiv
Neurol Neurosurg 1:13–14 Gabrielsson L, Mattsson S, Fowler C (2016) Palmitoylethanolamide for
the treatment of pain: pharmacokinetics, safety and efficacy. Brit
J Clin Pharmacol 82:932–942. https://doi.org/10.1111/bcp.13020 Gabrielsson L, Mattsson S, Fowler C (2016) Palmitoylethanolamide for
the treatment of pain: pharmacokinetics, safety and efficacy. Brit
J Clin Pharmacol 82:932–942. https://doi.org/10.1111/bcp.13020 Del Re A, Corpetti C, Pesce M, Seguella L, Steardo L, Palenca I, Rurgo
S, De Conno B, Sarnelli G, Esposito G (2021) Ultramicronized
palmitoylethanolamide inhibits NLRP3 inflammasome expression
and pro-inflammatory response activated by SARS-CoV-2 spike
protein in cultured murine alveolar macrophages. Metabolites
11:592. https://doi.org/10.3390/metabo11090592 Ghazizadeh-Hashemi M, Ghajar A, Shalbafan M, Ghazizadeh-Hashemi
F, Afarideh M, Malekpour F, Ghaleiha A, Ardebili M, Akhondza-
deh S (2018) Palmitoylethanolamide as adjunctive therapy in
major depressive disorder: a double-blind, randomized and pla-
cebo-controlled trial. Declarations J Affect Disord 232:127–133. https://doi.
org/10.1016/j.jad.2018.02.057 g
j j
Gibbons C, Illigens B, Wang N, Freeman R (2010) Quantification of
sudomotor innervation: a comparison of three methods. Muscle
Nerve 42:112–119. https://doi.org/10.1002/mus.21626 Rejas J, Ribera M, Ruiz M, Masrramón X (2007) Psychometric proper-
ties of the MOS (Medical Outcomes Study) Sleep Scale in patients
with neuropathic pain. Eur J Pain 11:329–340. https://doi.org/10.
1016/j.ejpain.2006.05.002 p
Nerve 42:112–119. https://doi.org/10.1002/mus.2162 Hays R, Martin S, Sesti A, Spritzer K (2005) Psychometric properties
of the medical outcomes study sleep measure. Sleep Med 6:41–44. https://doi.org/10.1016/j.sleep.2004.07.006l Said G, Goulon-Goeau C, Slama G, Tchobroutsky G (1992) Severe
early-onset polyneuropathy in insulin-dependent diabetes mellitus. New Eng J Med 326:1257–1263. https://doi.org/10.1056/nejm1
99205073261905 Jin HY, Park TS (2018) Role of inflammatory biomarkers in diabetic
peripheral neuropathy. J Diabetes Investig 9:1016–1018. https://
doi.org/10.1111/jdi.12794 Sancho R, Calzado MA, Di Marzo V, Appendino G, Muñoz E (2003)
Anandamide inhibits nuclear factor-kappaB activation through
a cannabinoid receptor-independent pathway. Mol Pharmacol
63:429–438. https://doi.org/10.1124/mol.63.2.429i Juster-Switlyk K, Smith A (2016) Updates in diabetic peripheral neu-
ropathy. F1000Res, 5:738. https://doi.org/10.12688/f1000resea
rch.7898.1 Keppel Hesselink J, Kopsky D (2015) Palmitoylethanolamide, a neu-
traceutical, in nerve compression syndromes: efficacy and safety
in sciatic pain and carpal tunnel syndrome. J Pain Res 8:729–734. https://doi.org/10.2147/jpr.S93106 Schifilliti C, Cucinotta L, Fedele V, Ingegnosi C, Luca S, Leotta C
(2014) Micronized palmitoylethanolamide reduces the symp-
toms of neuropathic pain in diabetic patients. Pain Res Treat
2014:849623–849623. https://doi.org/10.1155/2014/849623 Solorzano C, Zhu C, Battista N, Astarita G, Lodola A, Rivara S, Mor
M, Russo R, Maccarrone M, Antonietti F, Duranti A, Tontini A,
Cuzzocrea S, Tarzia G, Piomelli D (2009) Selective N-acyletha-
nolamine-hydrolyzing acid amidase inhibition reveals a key role
for endogenous palmitoylethanolamide in inflammation. Proc Natl
Acad Sci USA 106:20966–20971. https://doi.org/10.1073/pnas.
0907417106 Koïtka A, Abraham P, Bouhanick B, Sigaudo-Roussel D, Demiot C,
Saumet J (2004) Impaired pressure-induced vasodilation at the
foot in young adults with type 1 diabetes. Diabetes 53:721–725. https://doi.org/10.2337/diabetes.53.3.721f Magrinelli F, Briani C, Romano M, Ruggero S, Toffanin E, Triolo
G, Peter GC, Praitano M, Lauriola MF, Zanette G, Tamburin S
(2015) The association between serum cytokines and damage to
large and small nerve fibers in diabetic peripheral neuropathy. J
Diabetes Res 2015:547834. https://doi.org/10.1155/2015/547834 Steels E, Venkatesh R, Steels E, Vitetta G, Vitetta L (2019) A double-
blind randomized placebo controlled study assessing safety, tol-
erability and efficacy of palmitoylethanolamide for symptoms of
knee osteoarthritis. Inflammopharmacology 27:475–485. Declarations https://
doi.org/10.1007/s10787-019-00582-9i Malik R, Tesfaye S, Newrick P, Walker D, Rajbhandari S, Siddique I,
Sharma A, Boulton A, King R, Thomas P, Ward J (2005) Sural
nerve pathology in diabetic patients with minimal but progressive
neuropathy. Diabetologia 48:578–585. https://doi.org/10.1007/
s00125-004-1663-5 The Expert Committee on the Diagnosis and Classification of Diabetes
Mellitus (2003) Report of the Expert Committee on the diagnosis
and classification of diabetes mellitus. Diabetes Care, 26:s5-s20. https://doi.org/10.2337/diacare.26.2007.S5 Malik R (2014) Chapter 18 - Pathology of human diabetic neuropathy. In: Zochodne DW, Malik RA (eds.) Handbook of Clinical Neurol-
ogy. Elsevier The International Expert Committee (2009) International Expert Com-
mittee report on the role of the A1C assay in the diagnosis of
diabetes. Diabetes Care 32:1327–1334. https://doi.org/10.2337/
dc09-9033 Mattace Raso G, Russo R, Calignano A, Meli R (2014) Palmitoyleth-
anolamide in CNS health and disease. Pharmacol Res 86:32–41. https://doi.org/10.1016/j.phrs.2014.05.006 Mu ZP, Wang YG, Li CQ, Lv WS, Wang B, Jing ZH, Song XJ, Lun
Y, Qiu MY, Ma XL (2017) Association between tumor necro-
sis factor-α and diabetic peripheral neuropathy in patients with
Type 2 Diabetes: a meta-analysis. Mol Neurobiol 54:983–996. https://doi.org/10.1007/s12035-016-9702-z Vas P, Papanas N (2020) Depression and diabetic peripheral neu-
ropathy: birds of a feather, but when do they flock together? Exp Clin Endocrinol Diabetes 128:347–349. https://doi.org/10.
1055/a-0808-4269 Yang J, Zhao Z, Yuan H, Ma X, Li Y, Wang H, Ma X, Qin G (2019)
The mechanisms of glycemic variability accelerate diabetic cen-
tral neuropathy and diabetic peripheral neuropathy in diabetic rats. Biochem Biophys Res Commun 510:35–41. https://doi.org/10.
1016/j.bbrc.2018.12.179 National Health and Medical Research Council (2016) Guidance:
Safety monitoring and reporting in clinical trials involving ther-
apeutic goods. National Health and Medical Research Council,
Canberra Ng F, Trauer T, Dodd S, Callaly T, Campbell S, Berk M (2007) The
validity of the 21-item version of the depression anxiety stress
scales as a routine clinical outcome measure. Acta Neuropsy-
chiatr 19:304–310. https://doi.org/10.1111/j.1601-5215.2007.
00217.x Zelman D, Gore M, Dukes E, Tai K, Brandenburg N (2005) Valida-
tion of a modified version of the brief pain inventory for painful
diabetic peripheral neuropathy. J Pain Symp Manag 29:401–410. https://doi.org/10.1016/j.jpainsymman.2004.06.018 Zochodne D (2014) Chapter 26 - Mechanisms of diabetic neuron dam-
age: Molecular pathways. In: Zochodne DW, Malik RA (eds.)
Handbook of Clinical Neurology. Declarations Elsevier Paladini A, Fusco M, Cenacchi T, Schievano C, Piroli A, Varrassi G
(2016) Palmitoylethanolamide, a special food for medical pur-
poses, in the treatment of chronic pain: a pooled data meta-anal-
ysis. Pain Phys 19:11–24 Publisher's Note Springer Nature remains neutral with regard to
jurisdictional claims in published maps and institutional affiliations. Publisher's Note Springer Nature remains neutral with regard to
jurisdictional claims in published maps and institutional affiliations. Patel S, Santani D (2009) Role of NF-κB in the pathogenesis of diabe-
tes and its associated complications. Pharmacol Rep 61:595–603. https://doi.org/10.1016/S1734-1140(09)70111-2 1 3
|
W2090205417.txt
|
https://zenodo.org/records/1968983/files/article.pdf
|
de
|
Zur Prophylaxe der Diphtherie
|
Medical microbiology and immunology
| 1,901
|
public-domain
| 9,580
|
Zur l~rophylaxe der Diphtherie.
Yon
Doc. G. Gabritschewsky,
Vorstand des bakterio|ogisohen Institutes an tier Kaiserl. Universit~.t ztl Moskatu
Unsere heutigen Kenntnisse fiber die Diphtherie haben dank den
Arbeiten hervorragender Bakteriologen, wie LSffler, Behring, Ehrlich,
Roux u. A. eine solche Ausgiebigkeit erlangt, dass uns eine gleiche ffir
viele andere acute Infectionskrankheiten erwfinsoht whre. Die Bakteriologie hat uns nicht nur die Aetiologie, Pathogenese und Therapie der
Diphtherie klar gestellt, sondern sie gab uns auch werthvolle Mittel in
die Hand ffir die Prophylaxe dieser Krankheit.
Indem wir die Frage aber die Pr~ventivimpfungen mit specifischem
Serum bei Seite lassen, wollen wit die in der Litteratur niedergelegten
bakteriologischen Angaben i~ber das Auffinden yon Diphtheriebacillen im
Rachen der Reconvalescenten und Gesunden sowie der hieraus zu folgernden prophylaktischen Maassnahmen analysiren. Schon im Jahre 1884
wies LSffler zuerst auf die MSglichkeit des Antreffens von Diphtheriebacillen auf der Schleimhaut Gesunder bin.
Im Jahre 1890 wurde auf dem internationalen medicinischen Congress
zu Berlin im Namen des Dr. Roux (I) ein Referat, in welchem folgende
-~[aassnahmen zur Bek~mpfung tier Diphtherie empfohlea wurden, verlesen. Roux verweist auf die Nothwendigkeit einer friihzeitigen Diagnose
tier Diphtherie auf bakteriologischem Wege, h~lt das Isoliren der Reeonvalescenten bis zum Schwund der Bacillen aus der MundhShle ffir erforderlieh, sowie das hhufige Untersuehen der Halsorgane yon Schfilern,
besonders wenn unter den letzteren F~lle yon Diphtherie vorgekommen etc.
Nachdem F e e t (2) im Jahre 1893 sieh iiberzeugt hatte, dass die Diphtherie auch als Angina oatarrhalis, mit Fieber, aber ohne Membranen~
46
~. ~ABRITSCHEWSKY:
auftreten kSnne, wandte er die bakteriologische Untersuchung behufs einer
friihzeitigen Diagnosenstellung bei allen Kindern sowohl mit Anginen, als
auch bei gesunden, die einer m5glichen Infection unterliegen konnten, an.
Auf dem medicinischen Congress zu Budapest, im Jahre 1894, wurde
auf das nothwendige Isoliren Diphtheriekranker bis zum vollstiindigen
Verschwinden des Infec~ionsstoffs yon den Schleimh~uten sowie auch, wo
das nur mSglich, auf das Untersuchen yon Gesunden, welche mit Diphtherieherden in Berfihrung gekommen, hingewiesen.
Ich will bier nur einige der 18 Thesen, welche auf dem Budapester
Congress yon der deutschen Commissioil, die aus den Professoren LSffler,
Behring, hIosler, P i s t o r und Strfibing (3) bestand, in Vorschlag gebracht worden, anftihren :
,,Der Diphtheriebacillus kann im Rachen bezw. in der Nase gesunder
Individuen vorkommen, ohne Krankheitserscheinungen zu machen" (aus
tier 5. These).
,,Der Kranke ist infectiSs, so lange er noch Bacillen auf den Schleimh~uten hat" (a. d. 7. These).
,,Reconvalescenten yon Diphtherie sind nicht eher zum freien Verkehr (Kinder zum Schulbesuche) zuzulassen, als bis durch die bakteriologische Untersuchung das Verschwinden der Bacillen konstatirt ist" - (a. d. 17. These).
Auf diesem Congress theilte. Dr. W. It. Welch viele interessante,
yon den amerikanischen Aerzten Big~,s, Park, Beebe und ihm selbst
(4) gesammelte Thatsachen mit.
Von 752 bakteriologisch yon amerikanischen Collegen untersuchten
Diphtherief~llen konnten dieselben bei 325 Kranken 3 Tage nach dem
Verschwinden der Membranen keine Bacillen mehr nachweisen; bei 201
5 bis 7 Tage; bei 84 nach 12 Tagen; bei 69 nach 15 Tagen; bei 55
nach 3 Wochen, bei 11 nach 4 Wochen, bei 15 nach 5 Wochen und
bei 1 nach 7 Wochen liessen sich keine Bacillen mehr nachweisen.
Die bakteriologische Untersuchung yon 330 !ndividuen, welche in
keinem directen Verkehr mit Diphtheriekranken gewesen, ergab bei 8 yon
ihnen virulente Diphtheriebacillen. Zwei yon diesen letzteren acht erkrankten sparer an Diphtherie. In 14 Familien mit 48 Kindern, bei
welchen die Isolirung der Diphtheriekranken mangelhaft oder gar nicht
durchgeffihrt worden war, land man bei 50 Procent der Kinder im Raehen
virulente Diphtheriebacillen. 40 Procent dieser inficirten Kinder erkranl~te
an Diphtherie. Andererseits konnte man bei vollkommenem Isoliren nut bei
10 Procent der gesunden Kinder Bacillen nachweisen.
Auf dem medicinischen Congress zu Bristol wies in demselben Jahre
(1894) Dr. tI. Biggs (5) darauf hin, dass, nachdem das Gesundheitsamt
ZUR PEOPHYLAXE DEE DIPHTHERKE.
47
zu New-York die bakteriologische Untersuchung ffir auf Diphtherie verd~chtige Fille organisirt, die Zahl dieser letzteren auf 5- bis 6000 gestiegen. Diese Untersuchungen ergaben dem Gesundheitsamt, dass das
Weiterverbreiten der Krankheit hauptsSchlich dutch die Reconvalescenten,
bei denen selbst 7 bis 8 Woehen im Rachen noch virulente Bacillen angetroffen werden, vor sich gehe. Darauf bin besehloss das Gesundheitsamt, Reconvaleseenten nur dann als gesund zu be~achten und die Desinfection der Riiumlichkeiten zu gestatten, wenn die bakteriologische Untersuchung in Bezug auf Diphtheriebaeillen negativ ausgefallen.
Im Allgemeinen muss man sagen, dass die Bekhmpfung der Diphtherie nirgends so rationell gehandhabt wird wie in New-York und anderen
amerikanisehen Stiidten. 1 0hne hier n~her auf das Ausffihren der bakteriologisehen Untersuchung bei Diphtherie einzugehen, gestatte ich mir
nur den Hinweis auf prophylaktisehe Haassnahmen (Isoliren, Desinfeetion),
in Bezug auf welche alas New-Yorker Gesundheitsamt alles, was nur heut
zu Tage mSglich ist, anwendet. Als bester Beleg hierffir kSnnen die
Circul~re und Instructionen fiir Aerzte und das Publicum dienen, die mir
mit vielem anderen Material zur Bekimpfung der Diphterie in liebenswfirdiger Weise yon Herrn Dr. H. M. Biggs, Vorstand der bakteriologischen Station am Gesundheitsamt-zu New-York zur Verffigung gestellt
wurden und die ich anhangsweise zu meiuer Arbeit wiedergebe.
Im Jahre 1894 erschien eine sehr ausffihrliche Arbeit fiber die Epidemiologie der Diphtherie yon Professor C. Flfigge (6), in welcher tier Verfasser
unter Anderem sich dahin iussert, dass das Weiterverbreiten der Diphtherie hauptsiehlich durch Gesunde (Reconvalescenten) statthabe. Aeussere
Einfiiisse, terrestrische, klimatische u. s. w. sind yon untergeordneter Bedeutung, wogegen Lebensalter und sociale Einflfisse, enges Zusammenwohnen, reger Verkehr zwischen Inficirten und Gesunden eine gosse
Rolle spielen.
In demselben Jahr (1894) haben L5ffler und Abel (7) eine bakteriologische Untersuchung yon gesunden Schiilern w~thrend einer Diphtherieepidemie vorgenommen. Bei 4 yon 160 Schiilern fanden sich Diphtheriebacillen. Zwei yon diesen 4 erkrankten spiter an Diphtherie.
1894 stellte Dr. Aaser bakteriologische Massenuntersuchungen in
einer Cavalleriekaserne in Christiania nach wiederholten DiphtheriefSllen
im Laufe yon 4 Monaten an, trotzdem dass die Kranken jedesmal unverztiglich ins Hospital untergebracht und die Wohnriume, als auch die
Kleider desinficirt wurden. Die Untersuchung ergab, dass yon 89 Individuen 17, i. e. 19 Procent, virulente Diphtheriebacillen beherbergten.
1 Vgl. K o l l e .
Diese Zeitschrift. Bd. XIX,
48
~. ~ABRITBCHEWSKY:
Alle inficirten Soldaten wurden gleich darauf isolirt. Einer yon ihnen
erkrankte am niichs~folgenden Tage an einer schweren Diphtherie; bei
zwei anderen trat spiite~ eine Angina lacunaris mit unbedeutender Membranbildung und Fieber auf. Bei den fibrigen kam es nur zu einer
Hyper~mie der Rachenschleimhaut, die so lunge anhielt als die Schleimhaut Diphtheriebacillen beherbergte. Der gfinstige Erfolg dieser ~aassnahmen - - das ErlSschen der Diphtherie-Epidemie in der Kaserne - - ,
bemerkt der Autor, veranlasste ihn die bakteriologisehe Untersuchung
auch in einigen anderen F~illen zu verwerthen. Als in einer Scharlachabtheilung ein Fall yon Diphtherie auftrat, ergab die Untersuchung aller
Kinder dieser Abtheilung bei 20 Procent yon ihnen Diphtheriebacillen.
Naehdem alle infieirten Kinder, die in eine andere Ab~heilung fibergefiihrt
worden, Pr~ventivinjectionen erhalten hat~en, traten keine neuen Erkrankungen aufi
Interessant ist der Umstand, dass eine dieser Kranken, bei welcher
veto 6. bis 31. October Diphtheriebaeillen noch nicht verschwunden
waren, nach Hause entlassen wurde. Am 6. November wurden' zwei
kleine Schwestern dieser entlassenen Kranken ins Spiral mit Diphtherie
eingeliefert.
Ein anderes ~Ial kam wiederum in derselben Scharlaehabtheilung ein
Fall yon Diphtherie vor; bei der bakteriologischen Untersuchung yon
29 Kindern constatirte man bei 9 virulente Diphtheriebacillen. Bei einem
der isolirten Kinder stell~e sich eine leichte Diphtherie - - ]llembranen auf
den Tonsillen - - ein.
In demselben Jahre 1894 empfahl D e s c h a m p e s (9) auf Grund der
Thatsachen, dass die Diphtherie dutch Reeonvalescenten und durch zu
frfih aus den Hospitiilern Entlassene in die Familien eingeschleppt werde,
das Erriehten yon St~itten ftir Diphtheriereeonvalescenten.
Auf Grund der Untersuchungen fiber die antitoxischen Eigensehaften gesunder Individuen dem Diphtheriegift gegenfiber kam W a s s e r m a n n (t0)
im Jahre 1895 zum Schluss, dass viele gegen Diphtherie refract~ire Individuen auf ihren Schleimhauten lange Zeit hindurch virulente Diphtheriebacillen beherbergen und somi~ die Diphtherie weiter verbreiten kSnnen.
Von 20 Kindern, die der MSglichkeit einer Infection unterlagen, fand
Wassermann b e i 3 yon ihnen Diphtheriebacillen; eins dieSer letzten drei
Kinder erkrankte am niichsten Tag an Diphtherie.
Indem W a s s e r m a n n auf die bakteriologisehe Untersuchung als auf
eine prophylaktische Haassnahme bei Bekiimpfung der Diphtherie hinweist;
spricht er sieh dahin aus, dass die Untersuchung nieht nur bei Erkrankten
ausgefiihrt, sondern auch auf die seheinbar Gesuuden ausgedehnt werden
mtisse. Die Umgebung der Kranken kann im Sinne eines Weiterver-
49
ZuR 1)ROI~HYLAXE DER Drl)HTH~RIE.
schleppens der Infection nur dann als ungef~hrlich betrachtet werden,
wenn die bakteriologische Uutersuchung negativ ausgefaUen. Ein besonderes Augenmerk ist darauf zu richten, dass den Geschwistern erkrankter Kinder nicht frfiher der Schulbesuch gestattet werde, bis dutch
die Untersuchung des Rachens die Anwesenheit yon Diphtheriebacillen
sichergestellt ist.
Diese Vorschrift ist noch nach anderer Richtung bin yon einschneidender Wichtigkeit, weil sonst die Desinfection der Wohnriiume oft v511ig
illusorisch wird.
Auf den XIII. Congress ffir innere Medicin zu Mfinchen im Jahre
1895 theilte Dr. Trump (11) die Resultate seiner bakteriologischen Untersuchungen fiber Diphtherie mit, auf Grand deren er zu folgenden
Schlfissen kommt :
1. Bei Kranken mit Angina diphtherica finder man oft Diphtheriebacillen auch auf anderen Schleimhhuten, ohne dass irgend welche pathologische Verhnderungen an denselben vorliegen.
2. Reconvalescenten beherbergen Diphtheriebacillen zuweilen sehr
lange (in einem Falle konnte man dieselben noch 82 Tage na~h tier Erkrankung nachweisen); bei gesunden Kindern liessen sich Diphtheriebacillen konstatiren, die wahrscheinlich dutch Reconvalescenten aus den
Diphtherieabtheilungen eingeschleppt worden waren.
3. Gesunde Individuen, die Bacillen beherbergen, kSnnen die Quelle
yon Hausepidemieen werden.
4. Die Infection steht auf dem Wege des directen Contactes; weshalb
eine blosse Desinfection der Wohnriiume und Effecten zum Yerhiiten einer
Weiterverbreitung der Infection sich als ungeniigend erweist.
Im Jahre 1896 untersuchte Dr. Mfiller (12) in der Klinik des Professors H e u b n e r systematisch alle Kinder, wobei yon 92, welche vom
Betreten his zum Verlassen des Hauses beobachtet wurden, 20 virulente
Diphtheriebacillen aufwiesen. Der Autor vermerkt die wichtige Bedeutung
solcher Untersuchungen fiir die Prophylaxe tier Diphtherie, da gegen
diese Infection immune Individuen ,wandernde Diphtherieherde" abgeben
kSnnen.
In demselben Jahr sprechen sich Kewlett, Nolan (13) and G u n d o b i n
(14) dahin aus, dass nur die bakteriologische Untersuchung als Kriterium
ffir die Isolationsdauer der Kranken yon Gesunden heranzuziehen sei.
Im Jahre 1896 spricht 1~'. H. Williams (15) auf Grund seiner
Beobachtungen sich dahin aus, dass in Famflien, in welchen die ersten
Diphtherief~lle aufgetreten, es nothwendig ist eine bakteriologische Untersuchung auch der scheinbar Gesunden vorzunehmen. Ffir den sicheren
Zeitschr. L Hygiene. XXX~'I.
4
5O
~. ~ABRITSCHE~'SK~ :
Nachweis yon Diphtheriebacillen im Rachen ist zuweilen ein einma]iges
Untersuchen nieht hinreichend.
Im Jahre 1897 legte C. F r a e n k e l (16) in einer sehr interessanten
Arbeit diejenigen allgemeinen Grundlagen zur Bekfimpfung der Diphtherie,
die auf dem heutigen Stand der Wissenschaft basiren, nieder. Er stellt
als ein nothwendiges Postulat eine rationellere Bekampfung der Diphtherie
auf als das bisher geschehen und weist auf die Grundlosigkeit der Gegner
und Skeptiker gegen die Maassnahmen zur Bek~mpfung der Diphtherie
bin. C. F r a e n k e l betrachtet als die wiehtigste Quelle der Infection den
Menschen (K_ranken, Reconvalescenten and Gesunden) und von untergeordneter Bedeutung versohiedene Effecten, die mit den Sekreten der Schleimh~iute Inficirter verunreinigt sind: Desinfection und Isolation sind somit
die besten und rationellsten Mittel. Da das Isoliren yon Kranken in
Privathiiusern, ja selbst in reichen, auf grosse Schwierigkeiten stSsst, so ist
es nothwendig, mSglichst friih die Kranken ins Hospital zu sohaffen. M_it
der Heilung der Krankheit selbst ist die Aufgabe des Klinikers beendet,
da aber der Reconvalescent zuweilen noch lange Zeit Tr~ger der Diphtheriebacillen ist, so beginnt jetzt die Aufgabe des Hygienisten und Sanit~tsarztes, dem Weiterversehleppen der Infection vorzubeugen. Es muss
somit als ein logisches Postulat ffir die Prophylaxe anerkannt werden ,,das
Errichten an den ttospit~ilern yon Stationen, in welchen alle diejenigen
Reconvalescenten, die noch L5ffler'sche Bacillen beherbergen, zuriickbehalten werden".
Eine strenge Isolirung ist nioht nur ffir Fiille yon sehwerer wirklicher
membranSser Angina diphtherica, sondern aueh fiir die leichteren Forn~en
derselben, ffir verschiedene andere durch Diphtheriebaoiiien veranlasste Infectionen, sowie end]ich auch ffir gesunde Individuen, die zuweilen Tr~ger
tier Infection sein kSnnen, nothwendig. ,,UnmSglicheAnforderungen", werden
Viele yon Ihnen ausrufen. Ganz richfig, erwidert der Autor selbst auf
diesen Einwurf; heut zu Tage ist die Realisirung dieser Wfinsche im
ganzen Umfange kaum mSglich, aber ffir die Praxis ist es nothwendig
die Aufgabe zu stellen und das Ziel, das Schritt fiir Schritt in rastloser
Arbeit erreicht werden sell, zu bestimmen. An und fiir sich UnmSgliohes
verlangen die vorher aufgestellten Si~tze gewiss nicht.
Im Jahre 1897 wies F i b i g e r (17) auf Grund der einsohl~igigen
Litteratur und durch eigene Beobachtungen nach, dass das Weiterverbreiten
tier Diphtherie dutch gesunde, abet infieirte Individuen statthabe und
besehrieb ausfiihrlich eine Diphtherieepidemie. in welcher er bakteriologische
Untersuchungen und Isolation der Gesunden - - das A as er'sche Verfahren
zur Bekiimpfung der Diphtherie, wie er dasselbe bezeichnet - - anwandte.
ZUR PRO]~HYLAxE DER DIPRTHERIE.
51
Bevor F i b i g e r auf die Beschreibung der yon ihm beobachteten Fiille
eingeht, giebt er Zahlenangaben fiber die Diphtheritieepidemie in Stockholm, wo Dr. t t e l l s t r S m zur BekSmpfung der Epidemie das Isoliren der
Kranken yon Gesunden durchffihrte, an.
I n den Jahren 1894--1895 beobachtete t t e l l s t r S m in einem Leibgarderegiment 25 Diphtherieffille. Da das Ueberfilhren der Kranken ins
Spiral und wiederholt ausgeffihrte Desinfectionen die Epidemie nicht zum
Stillstand brachten', so schritt man zur Untersuchung yon allen 786
Soldaten, unter welchea man bei 151, also in 19,21 Procent virulente
Diphtheriebacillea konstatirte. Alle Inficirten isolirte man yon den Gesunden; nur einer yon den Inficirten erkrankte sp~iter an Diphtherie.
Nach diesen Maassnahmen erlosch die Diphtherieepidemie in diesem Regiment.
F i b i g e r selbst beschreibt eine Diphtherieepidemie in einem Gymnasium, wo 8 Schiller in zwei Zeitabschnitten mit einem Interval yon
32 Tagen, w~hrend weichen die Schiller anlhsslich tier Feiertage entlassen
worden waren, erkranktefi. Als die gew5hnlichen Maassnahmen --~ Isolation
und Desinfection - - zu Nichts gefilhrt, entschloss sich F i b i g e r alle
134 Schiller, wie auch das Dienstpersonal am .Gymnasium zu untersuchen,
wobei in 22 Fallen Pseudodiphtherie-, fragliche Diphtherie- und echte
Diphtheriebacillen (10 Ffille) konstatirt wurden. Nachdem man die Inficirten isolirt hatte, kam 1 1/2 Jahre lang kein neuer Fall vor.
Interessant sind einige Angaben fiber die Dauer des Vorhandenseins
von Diphtheriebacillen im Rachen. Bei einem Knaben verblieben virulente
Diphtheriebacillen im Rachen 4 Monate lang, in einem anderen Fall selbst
9 Monate. Mit Rficksicht auf solche Ausnahmsfiille weist F i b i g e r aus
der einschl5gigen Litteratur auf den Fall yon Le Geudre et Pochon (18)
hin, in welchem 1 1/~ Jahre nach Ablauf des diphtheritischen Processes
noch Bacillen konstatirt werden konnten.
Auf Grund der soeben angeffihrten Thatsachen steht es i'fir F i b i g e r
fest, dass das Isoliren der Inficirten und die Desinfection im Princip das
allerbeste Mittel zur BekSmpfung yon Diphtherieepidemieen in Anstalten,
wo in der Masse der dort zusammenkommenden Menschen die allergilnstigsten Bedingungen filr ein Weiterverbreiten der Infection vorliegen,
ist. Gleich C. F r a e n k e l v.erhehlt auch F i b i g e r sich nicht die Schwi~rigkeiten bei der praktischen s
dieser Maassnahmen zur Bek~mpfung
der Diphtherie und meint, dass heut zu Tage ein e ideale Isolirung ,~ller
Inficirten i m ganzen Umfange nicht durchfilhrbar wi~re. Die yon praktischer Seite erhobenen Einwfirfe gipfeln 1. in der Schwierigkeit-alle
Inficirten zu entdecken, haupts~ichlich bei einmaligem Untersuchen u n d in
denjenigen F~llen, in welchen nut spSrlich Diphtheriebaci]len vorhanden
52
G. ~ABRITSC~t~WsKY:
sind, und 2. in der NothwendigkeR Gesunde ziemlieh lange, zuweilen
selbst einige Monate zu isoliren.
,,Was aber thun," fragt der Autor, ,,wenn ein so langandauerndes
Isoliren unmSglich ist," und beantwortet selbst die gestellte Frage: dass
Alles vernaehl~ssigen und sieh selbst fiberlassen ist absolut verwefflich,
u m so mehr, wenn es sich (wie in Herlufsholm) um ein enges Zusammenleben junger Leute und sehr empf~nglicher Kinder handelt. F i b i g e r
giebt uns keine bestimmten praktischen Winke, wie dieses Dilemma zu
15sen w~ire: yon rationalen Maassnahmen zur Bekiimpfung der Diphtherie
abzusehen ist unmSglich, ihre Anwendung aber ~nsserst schwierig.
Im Jahre 1897 spraeh Gliicksmann (19) beim Hinweis auf das
Antreffen yon Diphtheriebaeillen bei vollst~ndig Gesunden Sich dahin aus,
dass solehe Individuen die Diphtherie verschleppen kSnnen, da sie die
Triiger und Zfichter des inficirenden Agens seien.
Auf Grund der uns bekannten Eigensehaften der Diphtheriebaeillen
behauptet der Autor, dass die Diphtheriebacillen sowohl yon Reeonvalescenten und Gesunden ebenso infeetiSs sind als aueh yon Kranken. ,iDas
Constatiren yon Diphtheriebaeillen bei Genesenden," f~hrt Gliicksmann fort,
,,ist ffir die Prophylaxe yon Bedeutung, da Reeonvales~enten nur dann
zum freien Verkehr zugelassen werden dfirfen, sobald sie keine Diphtheriebacillen mehr beherbergen."
Mit Rfieksicht darauf haben das Kinderspital und einige praktisehe Aerzte
in Ziirich in bestimmten Intervallen syst.~matisch jeden geheflten Fall so
lange'untersucht, bis die Diphtheriebacillen verschwanden. Ausnahmen
yon dieser Regel fanden nur bei Uebefffillung des Spirals oder auf dringendes Verlangen unbelehrbarer Eltern, die ihre Kinder frfiher bei sieh zu
Hause haben wollten, start.
In demselben Jahre (1897) ffihren F o u l e r t o n und L l e w e l l y n (20)
ein Beispiel einer Diphtherieinfection durch einen gesunden Knaben aus
einer Pension, in welcher Diphtheriefalle vorgekommen, an. Der Knabe
verliess die Pension am 30. XI., als alle Kranken nach Hause entlassen
worden waren, wurde zu Hause isolirt und erhielt eine W~rterin ftir sich
allein. A m 8. XII. und 13. XII. eonstatirte man im Rachen dieses Knaben
virulente Diphtheriebacillen, die sich am 15. XH., am 20.XII. und am
20. I. nicht mehr nachweisen liessen. In der Zeit, w~ihrend welcher der
Knabe in seinem Raehen Diphtheriebacillen beherbergte, infieirte er seine
Warterin, die am 21. XII. an Diphtherie erkrankte.
F o u l e r t o ~ und L l e w e l l y n empfehlen alle Sehiiler, die in die Schule
nach Sohliessen derselben in Folge einer Diphtherieepidemie zuriiekkehren,
bakteriologiseh zu untersuchen und nur die keine Diphtheriebaeillen beherbergenden Schiller zum Besuohe zuzulassen.
ZuR PROPHYLAXE DER DIPHTHERIE.
53
Im Jahre 1897 weist Dr, N e t t e r (21) in einer Arbeit, die mit der
Frage fiber das Isoliren yon acuten Infectionskranken sich besch~ftigt,
auf die Gefahr der Verbreitung der Diphtherie dutch Reconvalescenten,
in deren Rachen oft ziemlich lange Diphtheriebacillen angetroffen werden,
bin und spricht sich u. A. wic folgt aus: ,Leider giebt es bisher noch
keine Anstalten, in welchen man genfigend lange die aus den Pariser
Hospit~lern entlassenen Diphtheriekranken isoliren kSnnte."
In den
Pariser Krankenhhusern verbleiben die Inficirten gewShnlich nicht liinger
als 10 Tage in Folge Raummangels; das Anormale und Unerwilnschte
dieser Sachlage springt um so mehr in die Augen, als in Frankreich in
Bezug auf die Lehranstalten eine ministerielle Verfilgung yore Jahre 1893
besteht, nach welcher an Diphtherie erkrankte Schiller zum Besuch der
Schule erst nach Ablauf yon 50 Tagen zugelassen werden.
Im 5ahre 1898 beschrieben Simonin et B e n o i t (22) ihre Untersuchungsresultate fiber larvirte Diphtherie, yon welcher sie viele F~lle
dutch die bakteriologische Untersuchung in Cavallerieregimentern in Lyon
in den Jahren 1896 und 1897 feststellen konnten. Unter 108 Untersuchten hatten 9 vollkommen ausgesprochene Diphtherie und 23 larvirte
Diphtherie (12 mit Angina catarrhalis und 11 ohne dieselbe). Die Dauer
einer solchen Angina betrug 50 bis 40 Tage und in einem Falle selbst 55
Tage. Nach den Untersuchungen dieser Autoren kSnnen selbst bei Abwesenheit yon Membranen - - L~hmungen und wahrer Diphtheriemarasmus
sich einstellen. Was die Dauer des Verweilens yon LSffler'schen Bacillen
im Rachen betrifft, so wiihrt dieselbe in vollkommen ausgesprochenen
F~tllen yon Diphtherie im Durchschnitt 34 Tage, wogegen dieselbe bei
Angina diphtherica larvata 62 Tage und ohne Angina selbst 83 Tage betragt. Die larvirte Diphtherie pflegt man bei Leuten im vorgerfickten
Alter 5frets als bei jugendlichen Individuen und bei Kindern anzutreffen.
W~hrend der Diphtherieepidemie constatirte man larvirte Diphtherie in
22.3 Proc. und nach 3 Monaten in 18.18 Proc., w~hrend am nichtinficirten
Orte nur 3.84 Proc. sich nachweisen liessen. Treten diphtherieverd~chtige
F~lle unter einer ~enschenmasse (Truppentheilen) auf, so sollen auf Vorschlag der Autoren alle Individuen klinisch und bakteriologisch untersucht
und die Inficirten isolirt werden.
Im'Jahre 1898 sprach Dr. M u l e r t (23) sich dahin aus, dass in der
Prophylaxe der Diphtherie die Isolirung nothwendig und, soweit iiberhaupt mSglich, durchgefilhrt werden miisse. Die bakteriologische Untersuchung giebt in einer bestimmten Zeit ein sicheres Kriterinm zum Isoliren
tier Reconvalescenten. Dr. ~[ulert ffihrt einea im Jahre 1896 yon der
Medicinalabtheilung des deutschen Kriegsministerims erschienenen Erlass
an, in dem es heisst: Diphtheriereconvalescenten diirfen aus den La-
54
03. GABR1TS0HEWSK~':
zarethen nieht frfiher entlassen werden, als bis eine dreifache auf einander
folgende mikroskopisehe Untersuchung und die Cultur die Abwesenheit
der Diphtheriebacillen sieher stellt, naehdem der pathologisehe Process
kliniseh als tIeilung geendet. Leider, fiihrt Dr. Mulert fort, seheinen
Soiche Maassnahmen in unseren Civilhospitiilern nieht allgemein fiblich zu
sein, Was ffir die Prophylaxe der Diphtherie sehr erwfinseht W~ire, um so
mehr, als Wiederholte Beobachtungen sichergestellt haben, dass Recon:
valescenten nach Sehwnnd der ]~embranen und Ablauf des Fiebers noch
viete Woehen lang in ihrer RaehenhShle virulente Baeillen beherbergen
und das Entlassen soleher Patienten somit eine gefAhrliche Quelle ffir die
Weiterverbreitung der Diphtherie darstellt. Der Autor beschreibt eine
kleine Epidemie, in: weleher ein zu frfihes Entlassen eines Diphtheriekranken neue Fiille in der Familie derselben zur Folge hatte.
Mit Rficksioht auf den eben angeffihrten Fall weist Pros h t o s l e r
darauf hin, dass in seiner Klinik zu Greifswald diese Maassnahme nach
M5glichkeit durchgeftihrt wird und dass die ttauptschwierigkeiten in den
materiellen Verh~ltnissen gipfeln. Beide Autoren ~iussern ihr Bedauern
fiber die Abwesenheit gesetzlioher Bestimmungen betreffs der Zeitdauer fiir
die Isolirung yon Diphtheriekranken in den Hospit~lern.
Czaplewski (24) unterstreieht die Bedeutung der bakteriologisehen
Untersuehung ffir die Sieherstellung nicht nur der gewShnlichen, sondern
aueh der abortiven Formen der Diphtherie~ was ein genaues Bestimmen
der Quelle ftir die Weiterverbreitung der Infeotion und die Anwendung
entspreehender Maasnahmen ffir Isolimng und Desinfection ermSglicht.
In demselben Jahre (1898) kommt Grigorjew (25)auf Grund yon
bakteriologisehen Untersuchungen an Diphtheriereconvalescenten zu folffenden ScMfissen:
1. Eine bestimmte Frist ffir das Isolirtbleiben aller yon Diphtherie
genesender Kinder lhsst sich nicht festsetzen.
2. Der Verkehr soleher Kinder mit gesunden ist nur nach wiederholten bakteriologischen Untersuchungen des Sehleims aus Rachen-und
NasenhShle zu gestatten.
3. Eine solehe bakteriologisohe Untersuchung ist besonders nothwendig bei K!ndem in sohulpfliehtigem Alter.
4. Bei den Diphtherieabtheflungen in den Kospitfflern ist es noth,
wendig, fiir die Reeonvateseenten Krankens/ile, die naeh ~Sgliohkeit isolirt
yon denjenigen seien, in welchen die kranken mit frischer Diphtherie behafteten Kinder liegen, herzurichten.
Im Jahre 1898 stellten Vesbrook, Wison, ~fcDaniel, Adair (26)
eine systematisehe bakteriologisehe Untersuchung der ZSglinge einer Schule:
ZUl~ ~ROPI:[YLAXE DER DIPIITHERIE.
55
in welcher bei einer best~ndigen Zahl yon 250 Schfilern epidemische
Diphtheric seit 1897 ausgebrochen war, an. Von 478 untersuchten F~llen
war bei 306 ein negatives Resultat und yon den restirenden 172 mit
pusitivem l~esultat wiesen 68 Kinder die klinischen Symptome der Diphtherie auf, wogegen 104 gesund waren. Bei vielen Kindern konnte man
Bacillen noch im Verlaufe yon Monaten nachweisen. Auf Grund dieser
Untersuchungen kommen die Autoren zum Schluss, dass man an Diphtheric
erkrankte und mit ihr inficirte Kinder bis zum vollst~ndigen Schwund
der Diphthenebacillen bei zweimaligem negativem Untersuchungsresultat
isoliren mfisse.
Im Jahre 1898 publicirte Prof. S o e r e n s e n (27) seine im Laufe yon
zwei Jahren (1895 und 1897) angestellten Beobachttmgen an Scharlachkranken in Bezug auf die Anwesenheit yon Diphtheriebacillen nicht nur
in F~llen, in welchen irgendwelche klinische Symptome yon Seiten des
Kehlkopfes und der Trachea vorlagen, sondern auch nicht selten bei allen
Kranken, die in Abtheilungen, wo Diphtherief~lle vorkamen, lagen. Von
1500 Kranken hatten 240 (16 Proc.) Diphtheriebacillen and yon diesen
letzteren F~llen land m~n bei 32 (13-3 Procent) das klinische Bild
(hiembranen) der Diphtherie. Nur ein Fall ~:erlief letal.
Yon 213 Kranken mit Diphtheriebacillen erkrankten 5, nachdem
die bakteriologische Untersuchung positiv ausgefallen.
Von 1547 F~llen, die beim Eintritt in die verschiedenen Abtheilungen
des Hdspitals untersucht wurden, fanden sich bei 38 (2.5 Procent)l
Diphtheriebacillen vor, was eine der mSglichen Quellen ffir die Weiterverbreitung der Infection darbieten kann.
Leider finden sich in der Soerensen'schen Arbeit, auf welche viel
Mfihe verwendet worden, keine n~heren Hinweise in Bezug auf die
diagnostischen ~erkmale der Diphtheriebacillen.
Der Pseudodyphtheriebacillen ist mit keinem Wort Erw~hnung gethan, wenngleich die letzteren
so h/iufig auf den Schleimh~uten Gesunder und Kranker angetroffen
werden, als dass sic bei den Kranken Soerensen's gefehlt h~tten.
Die Virulenz der Calturen ist nur in sehr wenigen F~llen bestimmt
worden. Das Neisser'sche Verfahren konnte der Autor nicht anwenden,
da dasselbe erst im ffahre 1897 beschrieben wurde. Somit haben die angeffihrten Zahlen nur einen bedingten Werth und kann die Thatsache
1 v. R a n k e (M.iinchener reed. Woehenschrift, 1886, Nr. 42) land unter 94 F~llen
yon Scarlatina bei 65 Procent'Membranen, yon welchen bei 53.7 PrqeentDiphtheriebacillen eonstatirt wurden. Diese Untersuchunffen fallen ebenfalls in die Zeit, in
weleher die Differentialdiagnose zwisehen Diphtherie- und PseudodiphtheriebaciUen
nicht besonders exact durehgefiihrt wurde, was bei dem heu~igen Stand der Untersuchungsmethoden m5glich ist.
56
G. GABR1TSCHEWSKu :
des ~eltenen Befalienwerdens unter den Tr~gern der Diphtheriebacillen (?),
unabh~n~g yon allen anderen Umst~inden, nut dadureh erkl~rt werden,
dass die Zahl der 240 nicht nut F~lle mit echten Diphtheriebaoillen,
sondern auch solche mit Pseudodiphtheriebacillen umfasste.
Alle acumen F~lle yon Diphtherie und Triiger de, Diphtheriebaclllen
isolirte So erensen voa allen anderen Kranken dieses Krankensaals, das
Isoliren wurde dagegen nicht vorgenommen, wenn man Diphtheriebacillen
bei de, Mehrzahl oder bei allen Kranken des betreffenden Saales constatirte.
D u n b a r und Vogel (28) kommen auf Grund der einschlagigen
Litteratur zum Schluss, dass bei de, Bekiimpfung der Diphtherie die
allersicherste Combination yon Maassnahmen in tier Desinfection und
Isolirung, die dutch bakteriologisehe Untersuchung beg,finder, wie dies in
einigen amerikanischen St~idten durchgeffihrt wird, gipfelt.
Im vorigen Jahre sammelte Kober (29) alle bis auf ihn in tier
Litteratur niedergelegten Angaben fiber den Nachweis yon Diphtheriebaeillen bei Gesunden, wobei es sich herausstellte, class die bakteriologisehe
Untersuchung einen positiven Ausschlag bei 7 Proc. gesunden und solcher
Individuen, die mit Diphtheriekranken in keine Berfibrung gekomrnen,
und bei 18.8 Proc. solcher, die nachgewiesenermaassen einem mSglichen
Infieirtwerden unterIe~en hatten, ergab. De, Autor selbst eonstatirte unter
600 gesunden und solchen Individuen, die einer Infection yon Seiten Diphtheriekranker sich nicht ausgesetzt, Diphtheriebacillen bei 15, d. h. 2.5 Proc.
und yon 123 Personen, die aus Diphtherieherden stammten, Bacillen
bei 10, d. h. 8 Procent.
K o b e r behauptet, gestiitzt auf ein genaues Studium der einschliigigen
Litteratur so wie auch auf seine eigenen Erfahrungen, dass die Hauptgefahr in der Weiterverbreitung der Diphtherie somit yon Seiten dieser
scheinbar Gesunden drohe.
Ausserdem ffihrt de, Autor noch eine ganze Reihe yon interessanten
Thatsachen an, die ich bier fibergehe, um den Umfang meiner Arbeit
nicht zu sehr auszudehnen.
Ueber die Epidemiologie der Diphtherie sehreibt W eic h s elm a n n (30)
1899 wie folgt: eine wichtige Rolle in der Weiterverbrei~ung der Diphtherie spielt die oben angeffihrte Thatsache, dass Gesunde und seheinbar
Gesunde, sowie auch Reconvatescenten in ihren Nasen- und RaehenhShlen
virulente Diphtheriebacillen enthalten unct somit unbehindert die Keime
de, Krankheit weiterverbreiten kSnnen.
Im Jahre 1899 theflte .L. M a r t i n (31) die prophylaktischen Maassnahmen, die zur Bek~mpfung de, Diphtherie in drei kleinen St~dten,
Privas, Petit-Tournier und Flaviac angewandt wurden, mit. Unter allen,
prophylaktischen Maassnahmen, die man gegen die Diphtherie gebraucht
ZUR IDROPHYLAX/~ DEI~ DIPHTHERIE.
57
bemerkt u. A. der Autor, ist die einfachste die bakteriologische Untersuchung. Dieses Mittel gestattet ein frfihzeitiges Erkennen der Infection
und erweist sich ~usserst nfitzlich in den Schulen, indem es die Inficirten
zu erkennen und sie yon den Gesunden zu isoliren ermSglicht. Das Anwenden dieser Maassnahme ergab dem Autor in den ttospitiilern eben
solche vorzfigliche Resultate. Schon im Jahre 1895, als in die Diphtherieabtheilung eines der Pariser HospitSler mehrere Masernkranke iiberfiihrt
wurden, starben alle Masernkranke ungeachtet der Serumtherapie. M a r t i n
stellte alsdann eine bakteriologische Untersuchung aller masernkranker
Kinder an und fand bei 9 Diphtheriebacillen. •
dieser 9 isolirten
Kinder verstarben. Im Laufe yon drei Monaten wurden keine neuen Erkrankungen beobachtet.
Ein anderes Mal hSrte im Hospice des Enfants assist4s eine endemische
schwere Diphtherie erst dann auf, nachdem eine systematische bakteriologische Untersuchung der Kinder durchgefiihrt und die Inficirtea his
zum vollsthndigen Schwund der Diphtheriebacillen aus dem Rachen isolirt
wurden.
G a r r a t t und W a h l b o u r n (32), die im London Fever ttospital 666
Scharlaehkranke auf die Anwesenheit yon Diphtheriebaciilea im Rachen
untersuchten, eonstatirten bei 8 Kranken, d. h. in 1.2 Proc. Diphtheriebaeillen und bei 21~ d. h. in 3-2 Prec. Pseudodiphtheriebacillen. Diphtherie als Nachkrankheit des Scharlachs hSrte fast ganz auf, nachdem man
es sich zur Regel gemacht, einen jeden in die Seharlachabtheilung eintretenden Kranken bakteriologisch zu untersuchen und bei Anwesenheit
yon Diphtheriebacillen ihn sofort zu isoliren.
Bevor ich zu meinen eigenen Beobachtungen iibergehe, muss ich noch
auf zwei soeben erschienene Arbeiten hinweisen. In einem norwegischen
Seehospiz ffir tuberculSse Kinder, unter welchen Fhlle yon Diphtherie auftraten, stellte Dr. S in d in g- L a r s e n (33) als prophylaktisches Mittel bakteriologische Untersuchungen an. Trotz dieser Maasnahm% die wiederholt
ausgeffihrt, trotz Isolirung und wiederholter Desinfection hSrte die Diphtherie nicht auf. Der Autor gesteht dies selbst ein und verweist auf die
Unvollkommenheit in der s
der bakteriologischen Untersuchung,
wobei nur nach Beseitigung der Mhngel die Epidemie erlosch. Diese Art
der Bek:,impfung in der Privatpraxis stSsst auf viele praktische Schwierigkeiten, so dass nach Ansicht des Autors dieselbe nut flit Hospit~ler und
Asyle fiir tuberculSse Kinder, die nach seinen Beobachtunge n ffir diese
Infectionskrankheit ganz besonders disponirt sind, verwendbar ist. Zu
bemerken ist, class you der Zahl der mit schwerer Tuberculose Behafteten
i~l der angefahrten Epidemie 80 Procent erkrankten~ Yon denjenigen mit
leichter Tuberculose nut 16 Procent,
58
G. G.ABR1TSCHEWSKY:
Endlich finden wir in der Arbeit des Dr. C o b b e t t (34) dieselben
Anforderungen ffir eine rationell durchgeffihrte Prophylaxe der Diphtherie,
welcher alle Autoren beipflichteten, wie dies aus der einschl~gigen
Litteratur ersichtlich.
Der Autor glaubt, dass solche Maassnahmen behufs Isolirung Genesender und Gesunder bis zum Schwund der Diphtheriebacillen auf
praktische Schwierigkeiten stossen und zweifelt nicht, dass es zur Realisirung
derselben der Beihfilfe der Gesellschaft selbst bedfirfe. Mit Rficksicht auf
alles frfiher Angeffihrte ist es nothwendig, die Gesellschaft mit den neuesten
Ergebnissen der Wissenschaft fiber diese Frage bekannt zu machen sowie
darauf hinzuweisen, dass wir ~[ittel zur Bek~mpfung des epidemischen
Weiterverbreitens der Diphtherie besitzen. Da uns solche Maassnahmen
zu Gebote stehen, w~re es siindhaft, dieselben unbeachtet zu lassen.
Wenden wir uns jetzt zur Betrachtung derjenigen F~lle, in welchen
die bakteriologische Prophylaxe der Diphtherie bei uns in Russland angewandt wurde.
Aus dem medicinischen Bericht der Anstalten der Kaiserin ~1aria
ffir das Jahr 1897/1898 ersieht man, dass die auf bakteriologische Untersuchungen begrfindeten prophylaktischen Maassnahmen zur Bek~mpfung
der Diphtherie zuerst yon Prof. K. R a u c h f u s s scbon im Jahre 1896 in
einer Lehranstalt zu St. Petersburg zur Anwendung kamen, wogegen erst
am 28. Februar 1898 auf der medicinischen 'Conferenz dieses Ressorts
das yon Prof. K. R a u o h f u s s ausffihrlich zusammengestellte Project yon
Maassnahmen zur erfolgreichen Bek~mpfung der Diphtherie in geschlossenen
Anstalten gut geheissen wurde.
Das Projeot basirt auf folgenden zwei Thatsachen, die duroh den
Verlauf der Diphtherieepidemieen in den Petersburger Internaten best~tigt
wurden:
1. durch das Bestehen yon Anginae laounares
deutliche Anzeichen einer Diphtherie und
diphthericae
ohne
2. dutch das Antreffen yon virulenten Diphtheriebacillen bei gesunden
unempf~nglichen Individuen, die abet fiir die Infeotion empf~ngliche inficiren kSnnen.
Auf Grund dieser Thatsachen wurde besGhlossen, nicht nut die Erzieherinnen, welche an den verschiedenen Formen yon Angina erkrankten ,
sondern auch alle vollkommen Gesund% die in Infectionsherden lebten,
zu untersuchen und diese letzteren bei positivem Befund yon Diphtheriebacillen in eine besondere Abtheflung des Hospitals ,,fOr gesunde Infici~'ende" fiberzuffihren.
1897 brachte ich in Vorschlag, sich dieser Maassnahme in dem
ZUR ]~ROPHYLAY]~:DER ])IPftTHERIE.
59
adeligen Knabenpensionat zu Moskau zu bedienen, we in kurzer Zeit zehn
Diphtherief~lle vorkamen. Dr. W l a s s j e w s k i aus unserem Institut stellte
an 66 Schfilern, sowohl gesunden als auoh an Reconvalescenten Untersuchungen an und fand bei 21 Diphtheriebaeillen. Zwei yon ihnen erkrankten sphter an Diphtherie. Nach Isolirung der Infieirten und Desinfection erlosch diese Epidemie.
1899 hatte ieh selbst Oelegenheit, diese Maassnahme in einem der
Kinderasyle, we sich 11 Knaben im Alter yon 4 bis 7 Jahren und vier
Erwachsene befanden, durchzufiibren. Trotz eines sofortigen Ueberffihrens
der Erkrankten in ein Hospital und einer griindlichen Desinfection hSrten
die DiphtheriefSlle nioht auf.
Bei der Untersuchung aller Insassen ohne Ausnahme oonstatirte man
bei zwei Knaben, die angeblich an Diphtherie nicht krank gewesen, vimlente Diph~heriebacillen. Naohdem man die Inficirten yon den Gesunden
isolirt und eine gTtindliohe Desinfeotion erst bei negativem Bacillenbefund
der InScirtgewesenen ausgefiihrt, kamen neue Erkrankungsf~lle nicht
mehr vet.
Man wandte sich im vorliegenden Fall an reich, da alle fiblichen
~iaassnahmen zur Bek~mpfung der Diphtherie, drei Mal durchgeffihrt,
ganz resultatlos geblieben. Die Durchfiihrung tier bakteriologischen Maassnahmen stiess nicht im Entferntesten auf irgend welohe 8ohwierigkeiten
und Hindernisse, weder teohnischer- noch socialerseits.
In demselben Jahre (1899) hielt ioh auf dem Congress der Gesellschaft russiseher Aerzte zu Kasan einen Vortrag, in welchem ich die allgemeinen Grundlagen, die zu Gunsten einer bakteriologischen Untersuohung
nicht nur yon Reconvalescenten I sondern auch yon Gesunden als Maassnahme einer rationellen Prophylaxe zur Bekhmpfung der Diphtherie in
Vorschlag k~men, anfiihrte (35).
Zu Ende desselben Jahres (1899) trat in einem der hiesigen M~dcheninstitute Diphtherie auf; man sohritt aus prophylaktisohen Riioksichten
zu einer bakteriologischen Hassenuntersuohung aller im Institut lebenden
Sohfilerinnen. Die Resultate dieser ~[aassnahme sind yon Dr. B e r e s t new (36) verSffentlioht worden. I-Iier sei nur bemerkt, class im Laufe
eines Monats, ~r ~itte October bis ~itte November, 18 F~lle yon Diphtherie anf 230 Schfflerinnen uad 115 Personen -:- das Dienstpersonal und
dessert Familien ~ kamen. Von 15 Schfilerinnen, die auf Grund der
bakteriologisohen Untersuohung mit Diphtheriebacillen isolirt wurden, erkrankten 7, was auf die Exaotheit und N/itzliohkeit der Untersuchung
hinweist, da zweifelsohne nut wirkli~h Infioirte in's Lazarethiibergefiihrt
wurden, da durch die fr(ihzeitige isolirung tier Inficirten die wahrscheinfiche Weiterverbreitung tier Krankheit verringert wurde und endlioh eine
60
G. GABRITSCHEWSKY:
frfihzeitige Serdtherapie der Inficirten eingeleitet werden konnte. Es ist
somit gere.chtfertigt, anzunehmen, dass man den getroffenen Vorsichtsmaassregeln bis zu einem gewissen Grade auch das Ausbleiben yon Todesf~llen unter den 18 Erkrankungen zuschreiben kann. Da bei der Massenuntersuohung aller im Institut Lebenden (ca. 350 Individuen) am 2. und
3. November und sparer am 7. und 8. November es sich herausstellte,
dass auch ein Theft des Dienstpersonales inficirt war und andererseits bei
der grossen Extensit~it und Sehnel!igkeit, mit welcher die Epidemie sieh
weiter verbreitete, es an R~umlichkeiten flit die nSthige Isolirung fehlte,
so musste die Anstalt geschlossen werden.
In diesem Fall ergab die bakteriologische Untersuohung, dass die
Bekhmpfung der Diphtherie mit den gewShnlichen fiblichen l~Iaassnahmen
Desinfection und Isolation der Erkrankten - - vollkommen nutzlos gewesen und das Ziel bei Inficirtsein des Dienstpersonales unerreichbar geblieben war.
Im Januar 1900, also ca. 2 Monate sp~iter, kehrten die Schfilerinnen
in Partieen yon 30 bis 40 Personen in's Institut zurfick und unteflagen
einer bakteriologischen Untersuchung, wobei man bei 10 virulente Diphtheriebacillen antraf. Diese letzteren warden isolirt und aus dem Lazareth
erst nach S c h ~ n d tier Bacillen entlassen. Schfilerinnen mit Pseudobacillen wurde der Besuch der Schule gestattet. Neue Erkrankungsff~ille
kamen nicht mehr vor.
-
-
Ia Bezug auf diese Maassnahmen sagt Prof. F i l a t o f f (37) in seinem
Lehrbuche Folgendes: ,,In der Praxis werden solche l~Iassenuntersuzhungen
auf unfiberwindliche Schwierigkeiten stossen, z. B. in grossen Pensionen
und hauPts~chlich auf dem Lande."
Profi F i l a t o f f ffigt hinzu, (lass die Gefahr des Infir
auch
,,nicht so schreaklich ist, wie man annimmt".
Principielle Widersprfiche sehen wir in dieser Begutachtung nicht,
um so mehr, als der Autor selbst die Thatsache des •
von wirklichen Diphtheriebacillen bei Gesunden als feststehend betrachtet und bei
der Weiterverbreitung der Diphtherie dem anhaltenden Verweilen vi_rulenter Diphtheriebacillen auf den Schleimh~uten der Reconvalescenten eine
wichtige Rolle zuschreibt.
Profi F i l a t o f f entl~sst die Reconvalescenten bei Schwund ~ler Diphtheriebacillen noch vor Ablauf yon 3 Wochen, nach dieser Frist jedoch
unabhhngig vom Resultat der bakteriologischen Untersuehung.
Nach dem Circular der l~iedicinal-Departements in Russland werden
Schiller nach iiberstandener Diphtherie erst nach 3 Wochen zum Schulbesuch zugelassen. Der Zusatz zu dieser u
lautet: ,Wenn mSg-
ZUR ~ROPHYLA~:EDER DIPHTHERIE.
61
lich nach einer bakteriologischen Untersuehung - - nach Schwund der
L 5 ffl er' schen Bacillen."
Dieses Circular betrachte~ also eine dreiwSchentliche Frist ffir nicht
bindend sobald Diphtheriebacillen im Raohen lhnger angetroffen werden.
in der russischen Litteratur wird endlieh yon Dr. J a k o w l e f f
(St. Petersburg) auf Grund der bakteriologischen Untersuehung yon Diphtheriekranken, Reconvalescenten, sowie auch Gesunden u. A. angeffihrt,
dass yon 245 Gesunden, die mit Diphtheriekranken in Berfihrung gekommen, bei 16, d. i. in 6.5 Proeent, Diphtheriebacillen angetroffen
wurden.
Dr. J a k o w l e f f sohliesst sich auf Grund eigener Beobachtungen, sowie der in der Litteratur niedergelegten Angaben, vollkommen den yon
mir gemaehten Behauptungen an und best~tigt vollends, dass ohne bakteriologische U n t e r s u c h u n g eine e r f o l g r e i c h e B e k ~ m p f u n g der
Diphtherie u n m S g l i c h ist.
Das sind die Thatsachen und Meinungss
tier einze]nen
Autoren~ die ieh mSglichst vollst~ndig aus der einsehliigigen Litteratur
zu sammeln bemfiht gewesen, um zu zeigen, bis zu welehem Grade die
leitende Idee in der neuen Handhabung zur Bek~mpfung tier Diphtherie
~rztlicherseits Anerkennung und wie welt die neuen hIaassnahmen praktische Verwendung gefunden. Zu welch' einer allgemeinen Schlussfolgerung berechtigt dies?
Im vorliegenden Fall stSsst ein richtiger Schluss auf keinerlei
Sehwierigkeiten. Aerzte, Bakteriologen und Epidemiologen stimmen alle
dariu iibereia, dass Gesunde, Genesende und an Diphtherie nicht krank
gewesene Individuen nicht selten Tr~ger yon virulenten Diphtheriebacillen
in der Hund-, Nasen- und RachenhShle und die Hauptverbreiter der
epidemischen Diphtherie sein kSnnen, dass als rationelle prophylaktisehe
Maassnahmen die Desinfeet,ion und Isolirung, die nur bei bakteriologischer
Controle die gfinstigsten Resultate ergeben, betrachtet werden miissen,
und dass endlich das Erkennen des Diphtheriebacillus und die Isolirung
yon Gesundeu zwei der grSssten Sehwierigkeiten sind bei der exaeten und
systematischen Ausffihrung yon prophylaktischen Maassnahmen zur Bek~mpfung tier Diphtherie.
Betrachten wit nun diese Schwierigkeiten, da nut sie die praktische
Verwendbarkeit einer rationellen Prophy.laxe der Diphtherie, die auf einer
bakteriologischen Untersuehung begrtindet~ praeisiren.
Vorerst muss darauf hingewiesen werden, dass trotz aller Sehwierigkeiten, welche in jeder neuen und complicirten Saehe besonders markant
hervortreten, die in u
gebraehten Maassnahmen, wie aus der einschl~gigen Litteratur ersichtlich~ erfolgreich' bereits ausgeffihrt wurden.
62
~. ~ABRITSCH'EWSKY:
Es liegt somit kein Grund vor, behaupten zu wollen, dass die Ausftihrung
dieser ]~[aassnahmen selbst in so bevSlkerten 5ffentliehen Anstalten wie
Kasernen, Schulen, Asylen und Pensionen unmSglich ist. Wenn lman
abet die Rationabilit~t dieser Maassnahmen vollkommen berechtigt und in
einigen Fiillen durchffihrbar, trotz der bestehenden Schwierigkeiten, anerkennt, so fragt es sich, ob wir nieht verpfliohtet sind, diese prophy]aktischen Maassnahmen zur Bek~mpfung der Diphtherie welter auszuarbeiten
und die Schwierigkeiten mSglichst zu beseitigen.
Man daft den Umstand nieht ausser Acht lassen, dass trotz des
Serums und anderer therapeutischer ~Iaassnahmen die Diphtherie dennoch
viele 0pfer fordert und bei uns zu Lande (Russland) die Morbidit~t und
an einigen 0rten auch die Mortalit~it selbst zugenommen haben. So wies
Prof. Dr. R a u c h f u s s (38) nach, dass in Petersburg in den letzten Jahren
die h[orbidit~t um alas u
die hlortalit~it um das Dreifache gestiegen.
In Bezug auf die erste Schwierigkeit, d. h. das Erkennen des Diphtheriebacillus, sind wit auf Grund yon Beobaohtungen und Untersuchungen
in unserem Institut zu folgenden Szhlfissen gekommen:
1. Die Cultur muss auf eoagulirtem Serum aus dem Sehleim der
Raohen- und NasenhShle (besonders, wenn Schnupfen vorliegt) angelegt
werden.
2. Eine einmalige Untersuchung ist ffir den exacten Nachweis des
u
yon Diphtheriebacillen ungenfigend.
3. Ausser der iibliohen Tinction muss auch die Neisser'sehe, die
bei positivem Resultat die ~ISglichkeit giebt, echte Diphtheriebacillen
(virulente und niehtvirulente) sicher zu stellen, ausgeffihrt werden.
4. Ein vollst~ndig negatives Resultat bei Neisser'scher F~rbung
yon St~bchen, die morphologisch den Diphtheriebacillen iihnlich, weist iluf
einen pseudodiphtheritischen Charakter derselben bin.
5. In zweifelhaften F~llen, bei sehr geringer Anzahl yon Diphtheriebacillen, oder wenn dutch die Neisser'sche F~rbung n u t sehr wenige
Stfibehen tingirt werden, so entscheidet das Dilemma entweder eiue wiederholte Untersuchung der betreffenden Personen oder ein Ueberimpfen in
ein neues R5hrchen mit Serum, wo somit in der zweiten Generation
zahlreiehere Culturen yon Bacillen sioh einstellen und die Neisser'sche
F~irbung dieselben intensiver tingirt. Solch' einen Unterschied zwischen
der ersten und zweiten Generation beobachteten wit einige Mal; und dies
kann vielleicht (lurch einen geschwfichten Zustand der einzelnen Keime in
Folge eines Antagonismus der Bakterien und anderer physikalischer und
chemischer Einflfisse, wie sie auf der Schleimhaut vorzukommen pflegen,
erklhrt werden.
ZUR ~ROPHYLAXE D]~R DrPHTB~RrE.
6~
6. Wie wenig echte Diphtheriebacillen man auch antreffen mug, und
wenn dieselben bis zu eiuem gewissen Grade aueh ihre Virulenz eingebfisst
h~ben sollten, so unterliegen dennoch die sie beherbergenden Individuen
einer Isolirung und mfissen als Inficirte betrachtet werden.
7. Individuen mit Pseudodiphtheriebacitlen mfissen im Sinne der
Weiterverbreitung der epidemisehen Diphtherie als unge~hrlieh betrachtet
werden.
&us den angeffihrten Thesen ersieht man, dass wir eine wichtige
Bedeutung der Neisser' sehen F~rbung f~r die Differentialdiagnose zwischen
Diphtherie- und Pseudodiphtheriebacillen beimessen, was wit um so begriindeter thua, als fast alle Autoren, die sich mit dieser Frage beschbftigt,
dieselben Resultate erzielt. Zur Differenzirung des echten Diphtheriebacillus kbnnen ausser der Neisser'schen F~rbung noch die saure Reaction der Zuckerbouillon, die Virulenz und das Verhalteu dem specifischen
Serum gegenfiber herangezogen werden.
Wit glauben nicht fehl zu gehen, dass bei Anwendung dieser Untersuchungsmethode ffir das Erkennen der Diphtheriebacillen viele F~lle mit
Bacillen heut zu Tage nicht isolirt werden, welche yon friiheren Autoren
als Diphtheriebaeillen angenommen warden und somit die Durchf~hrung
des Isolirens gesunder Individuen bei weitem erleichtert wird.
Die Exactheit, mit welcher die bakteriologische Diagnose gestellt
werden kann, gestattet eiae auf der letzteren begriindete ttandhabung
dieser praktischen Maassnahmen. Ausserdem haben Beobaehtuugen ergeben, class bei diesem uncomplicirten technischen Untersuehungsverfahren
ein hierin erfahrener Arzt im Laufe yon 24 Stunden ohne Anstrengung
bis zu 50 Individuen bakteriologisch unte~suchen kann, selbst wenn man
yon jedem einzelnen zwei R5hrchen mit Culturen beschickt und aus einer
jeden yon ihr zwei mikroskopische Priiparate anfertigt. Bei Massenuntersuehungen ist selbstverst~ndlich der Kostenpreis bei weitem geringer, was
in praktischer Beziehung ebenfalls nicht unberiicksichtigt bleiben daft.
Da abet eine absolute Garantie unmbglich ist, dass alle F~lle mit geringen
echten Diphtheriebacillen dutch diese ]Hethode sichergeste]lt werden kbnnen,
so ist der Einwurf berechtigt, ob es sich aueh lohnt, die Isolirung vorzunehmen, wenn Diphtheriebacillen vielleieht nieht gefunden werden
kbnnen, oder die Mbglichkeit vorhanden ist, eine Pseudodiphtherie- fi~r
eine eehte Diphtheriecultur anzunehmen.: - Soleh' ein Uebersehen ist selbstverst~ndlich nicht erwfinscht, kann abet dessen ungeaohtet die bakteriologisehe Untersuehung nicht entwerthen, da die letztere jedenfalls ein
sicheres Mittel ~ebt, die Weiterverbreitung einer Epidemie bei weitem
einzud~mmen und eine rechtzeitige Serotherapie der Isolirten und Erkraukten gestattet. Wir wollen durchaus nicht die Bedeutung und Rolle
64
CT. GABRITSCHEWSKY:
der Diphtheriebacillen bei Gesunden untersch/itzen, andererseits aber die
Gefahr, welche dieser Umstand mit sieh bringt, auch nicht fibertreiben.
In praktischer Beziehung ist es lange nicht einerlei, ob auf 100 Personen
10 bis 20 oder nur 1 bis 2 mit Diphtherie Behaftete kommen; es ist
n~mlich zur Genfige bekannt, class nicht nur die Quaht/it, sondern aueh
die Quantitat des inficirenden Agens das Schicksal der Infection und Epidemie entscheidet. Uebrigens ist das angef~hrte Beispiel nur Eventuahtiit
und nicht Regel. So ist man z. B. auf Grund der Thatsache, dass Isoliren und Desinfection nicht in allen F~llen ihren Zweck erreichen, absolut nicht zum Schluss berechtigt, class man ihre Anwendung selbst
unter den obwaltenden Umstiinden unterlassen mfisse.
Die Hauptschwierigkeit in der Durchfiihrung der bakteriologischen
Prophylaxe w/ihrend einer Diphtherieepidemie gipfelt nicht in der Diagnose,
sondern in dem Umstande, dass virulente Diphtheriebacillen bei Gesunden
und Reconvalescenten zuweilen im Laufe yon vielen ~onaten mit einer
ungewShnlichen Hartn/ickigkeit sich halten. Alle bisher verwandten Desinfectionsmittel, selbst die st~rksten unter ihnen, z.B. das Subhmat:vermindern zwar did Zahl der Keime in loco wohin sie gelangen und kSnnen
alas Verschwinden der Diphtheriebacillen beschleunigen, doch kann man
nicht immer sicher sein, dass sie rasch genug verschwinden, da an einigen
Stellen der Schleimh/iute des Rachens und der Nase (in den SeitenhShlen)
die Vermehrung der Bacillen unbehindert fortdauern kann. Durch eine
Ausarbeitung der Desinfectionstechnik der Schleimhiiute gegeniiber der
Diphtherie mfisste die Bek/~mpfung der Diphtherieepidemieen bedeutend
erleichtert werden.
Es bleibt somit in roller Kraft die Frage, was zu thun ist, wenn bei
einigen Reconvalescenten und Gesunder~ Diphtheriebacillen I~onate lang
zurfickbleiben und ein weiteres Isoliren aus 5conomischen und socialen
Grfinden unmSglich wiM.
Wenn das Isoliren sich nicht durchffihren 1/isst und die daran
interessirten Personen auf dasselbe nicht eingehen wollen, so bleibt natfirlich nichts fibrig, als yon einer vollkommenen Anwendung dieser Haassnahmen abzusehen, hieraus folgt jedoch nicht, diese Mittel da zu unterlassen, wo sie ausffihrbar und zwar in dem Maasse, wie nur mSghch.
Selbst wenn ein vollstiindiges prophylaktisches Isoliren, besonders yon
Erwachsenen, unmSglich, so gestattet die bakteriologische Untersuchung,
welche die Quelle der Infection aufdeckt, denjenigen, die wenigstens bedingte Maassnahmen der eigenen Familie und der Gesellschaft gegentiber
durchzufiihren wfinschen, die nSthigen Mittel. Das hnwenden yon desinficirenden Gurgelw~ssern, das sorgfiiltige, ffir die Umgebung unseh~d]iche Vernichten der Excrete aus l~und und Nase, besonderes Geschirr u. s. w.
65
ZUR PROPHYLAXE DER DIPHTHERIE.
kann bis zu einem gewissen Grade die Gefahr der Weiterverschleppung
der Infection herabstimmen. In einem der anhangsweise welter folgenden
Circulhre des New-Yorker Gesundheitsamtes finden sich ausffihrlichere
Hinweise fiber die Art und Weise der Uebertragung yon Diphtherie~ aus
welchen ersichtlich, wie man die Umgebung vor einem Inficirtwerden
schfitzea kann, wenn ein Isoliren unmSglich.
LungentuberculSse, die mehr oder weniger intelligent und denen die
Gefahr ffir alle Anderen wohl bewusst, beobachten gewisse Vorsichtsmaassregeln, und wie wohl bekannt, strebt die Gesellschaft, der Tuberculose gegenfiber im grossen Maassstabe prophylaktische Maassnahmen
zur Verminderung und Vernichtung der Infectionsherde anzuwenden.
I.ieut zu Tage muss in Bezug auf die Diphtherie dieselbe Frage auf
die Tagesordnung kommen. Die Bakteriologie hat uns gelehrt, wie und
wo das Diphtherieagens nachgewiesen werden kann, wie auch, class Gesunde, Reconvalescenten oder an Diphtherie Nichterkrankte einen tfickischea
Herd ffir die Infection und eine der I-tauptursachen der epidemischeu
Weiterverbreitung der Diphtherie abgeben. Nur dutch eine ausgiebigere
Verwendung der bakteriologischen Untersuchung wird die Prophylaxe der
Diphtherie rational begriindet sein und die besten Resultate bei Bek~mpfung
der Diphtherieepidemieen ergeben.
Die prophylaktischen Maassnahmen zur Bek~mpfung der epidemischen
Diphtherie, welche auf bakteriologischer Grundlage beruhen~ sind yon mir
in Thesen bereits formulirt worden. 1
Berficksichtigt man die oben angeffihrten Litteraturangaben fiber diese
Frage, so liessen sich diese Thesen in etwas veriinderter und ergfinzter
Form, wobei Alles speciell Russland Betreffende weggelassen, wie folgt
formuliren:
1. Die bakteriologische Untersuchung des Schleims aus der ]~und-,
Rachen- uhd NasenhShle soll nicht nur behufs diagnostischer Zwecke an
Erkrankten, sondern auch aus prophylaktischen Grtinden an yon Diphtherie
Genesenden, sowie an Gesunden, die in diphtheritischenHerden sich aufhalten und einer Infection durch dieselben ausgesetzt gewesen sein konnten,
angestellt werden.
2. Inficirte Individuen unterliegen, unabh~ngig yon ihrem vollst~indigen Wohlsein, denselben prophylaktischen ]~aassnahmen (Isolirung und
Desinfection), wie Diphtheriekranke. Wo ein vollst~ndiges Isoliren unmSglich, mfissen diejenigen Maassnahmen, welche wenigstens das Weiterverbreiten der Infection (besonderes Geschirr, systematisches und ungefiihr1 Centralblalt fi~r Bakteriologie.
Zeitschr. f. Hygiene. XX:XYI.
1899.
Bd. X X V I .
Nr. 16/t7.
5
66
G'. GABRITSCHF.WSKY:
liches Vernichten der Excrete aus Nase und Mund, Desinfection der
Schleimhiiute u. s. w.) beschr~nken, angewandt werden.
3. Diphtheriekranke dfirfen nach erfolgter Genesung aus den Hospit~lern nicht vor Schwund der Diphtheriebacillen yon den Schteimh~uten
entlassen werden. Wenn in den Hospitiilern Platzmangel das Durchfiihren diese Maassnahme nicht gestattet oder die Eltern und Anstalten
die Entlassung ihrer Kinder und ZSglinge vor Schwund der Diphtheriebacillen fordern, so sollen Eltern und Anstalten fiber die drohende Gefahr
der Weiterverbreitung der Infection in Kenntniss gese~zt und ihnen gedruckte Instractionen fiber Vorsichtsmaassregeln eingeh~ndigt werden.
Bei Platzmangel in den Hospit~lern sollten Asyle ffir genesende Kinder,
sowie auch ffir Gesunde, welche in Diphtherieherden inficirt sind, errichter werden.
4. In den Kinderhospit~lern mfissen aufDiphtheriebacillen alle Kinder,
besonders yon Masern, Schar]ach und Tuberculose behaftete, untersucht
worden.
5. In Schulen, Asy]en, Pensionen und Familien, wo Diphtherie aufgetreten, soll eine Massenuntersuchung der Rachen-und NasenhShle ausgeffihrt und alle Inficirten im Verlaufe einer durch die bakteriologische
Untersuchung festgesetzten Frist isolirt werden.
6. Bei der Desinfer
der Wohnr~ume und Sachen muss das Resultat der bakteriologischen Untersuchung sowohl der Reconvaleseenten,
als auch der in Diphtherieherden Wohnenden berficksichtigt werden.
Anhang.
Nr. 1,
Health Department of the City of New York.
Criminal court building.
R e g u l a t i o n s r e g a r d i n g the isolation of cases of d i p h t h e r i a
in p r i v a t houses.
In private houses the duration of isolation of cases of diphtheria after
apparent complete convalescence of such cases shall be determined by the
physician in attendance, with the following conditions:
First. Children convalescent from diphtheria shall not be allowed under
any conditions to attend any kind of school, i. e., day school, sunday school,
dancing school etc., until cultures show the absence of diphtheria bacilli
from the throat.
Second. Circulars of informations regarding the persistence of virulent
diphtheria bacilli in the throats "of convalescent cases of diphtheria and the
ZUR PROPHYLA~:E DER DIPHTHERIE.
67
dangers from infections arising from such eases shall be furnished by the
Health Department and presented by the attending physician, if the patient
is a child, to the mother, father or guardian, or, if the patient i s an adult,
to the patient, and the meaning of the circular and the significance of its
contents explained to them.
Third. The attending physician, when continued isolation is not mainrained, shall immediately notify in writing the Chief Inspector of contagious
diseases of the Health Department, of his action, and desinfection of the
premises may them be performed by the Health Department.
Exceptions to this rule regarding the time of isolation of convalescent
cases of diphtheria in private houses shall be:
1. Teachers of all kinds.
2. Persons whose occupations bring them into immediate contact with
children.
For the special regulations of the Health Department regarding the
isolation of cases of diphtheria in boarding houses, apartment houses and
hotels, application should be made to the Chief Inspector o.f contagious diseases.
George B. F o w l e r , ~.D.,
Commissioner of Health.
By order of the Board of Health.
Emmons Clark,
Secretary.
C h a r l e s G. W i l s o n ,
President.
Nr. 2.
Health Department of the City of l~ew York.
Criminal court building.
C i r c u l a r of i n f o r m a t i o n r e g a r d i n g the d a n g e r of d i p h t h e r i a b e i n g
c o m m u n i c a t e d to o t h e r s by p e r s o n s who h a v e r e c o v e r e d f r o m
the d i s e a s e .
Diphtheria is due to a germ known as the diphtheria bacillus. This
germ is present in the membrane and in the secretions of the mouth, nose
and throat of cases of diphtheria. The disease is produced by the reception
of the germs into the mouth or nose of well persons.
The discharges from the nose and mouth of cases of diphtheria containing these germs, may be received on handkerchiefs, towels, bedclothing,
carpets, rugs, personal clothing, toys, books etc., may dry, become pulverised
and breathed in as dust, or the germs may be transmitted directly from the
sick to the well through personal contact; as in kissing the sick, and in the
use of drinking cups or eating utensils which have been employed by the
sick, or through the direct discharge of the secretions on to the hands, face
or clothing of the nurse, physician or attendat while making applications
to the nose or throat of the sick person. Thus, in numerous ways the
germs find their way from the throat of the sick to the mouth or air passages of the well, and if then the conditions are favorable for the develop5*
~. GABRITSCHEWSKY:
68
ment of diphtheria in the throat of the person receiving them, after a varying period the disease is produced.
Diphtheria only follows the reception of the germs in the throa~ if
favorable conditions for their growth exist thm'e. In many cases the germs
remain for many days, and increase in number, in the throats of healthy
persons who have been in contact with cases of diphtheria, without producing the disease. Well persons who have these germs in their throats m a y
convey them to others, who contract the disease, while they themselves
escape.
During convalescence from diphtheria the germs of this disease often
persist in the throat for many days after all signs of disease in the throat
have disappeared, and after the individual is entirely well. Investigations
show that in about twenty-five per cent. of cases they persist for three
weeks or longer after the beginning of the disease, in fifteen per cent
for four weeks or longer, in five per cent for five weecks and occasionally
even for a much longer time. Experiments have shown that almost invariably these germs are virulent and capable of inducing the disease in others,
so long as they persist in the throat, and persons having such germs in
their throats may convey the disease to other persons at any time. Observation, however, has proven that the chances of communication of the disease in well persons after complete convalescence are not great, excepting
among children, who are far more succeptible to the disease and who are
much more likely t o become infected through the frequent introduction of
infected articles into the mouth.
The danger of communicating the disease to others after recovery while
the bacilli persist, is less than during th~ disease, because the number of
germs is smaller, the secretions less abundant and not as likely to be discharged where they will gain entrance to the mouths and throats of other
persons. When persons do not remain completely isolated as they are strongly
advised to do-until cultures made from the throat show the absence of the
diphtheria bacilli-they should constantly remember the facts set forth in this
circular, and use every precaution to .prevent the communication of the disease through the secretions which incite the disease, and should be particulary careful in the relations and contact with children.
No child, convalescent from diphtheria, Will be allowed to return to
school or to attend any assembly of children until diphtheria bacilli are no
longer present in the throat, and no person engaged in teaching or in caring
for children, who is convalescent from diphtheria, will be allowed to resume
his or her occupation until the bacilli are no longer present in the throat.
G e o r g e B. F o w l e r , M.D.
Commissioner of Health.
By order of the Board of Health,
E m m o n s Clark,
Secretary.
C h a r l e s C. Wilson,
President.
ZUR PROPYIY:LAXE D:ER DIPHTHERIE.
69
Litteratur-Verzeichniss.
1. Deutsche reed. VFochenschrift. 1890. Nr. 46.
2. CorresTondenzblattfis
1893. Bd. X X [ I I . Ref. Centralblair fi~r Bakterlologie. Bd. XIu
3. Centralblatt fi~r Bakteeiolo.qie. Bd. XVI.
4. The American Journal of ~he reed sciences. October 1894. - - Ci~irt nach
C. F r a e n k e l .
5. British reed. Journal. 1894. Vol. II. p. 360.
6. JOiese Zeitsehrift. Bd. XVII.
7. Deutsche med. BToehensehrift. 1894. Nr. 47.
8. JEbenda. 1895. Nr. 22.
9. Revue d'h.Tgi$ne et de police sanitaire. T. XV. p. 241.
10. Diese Zeitsehrifl. Bd. XIX.
11. Deutsche reed. ~ochenschrift. 1895. Nr. 10. S. 68.
12. Jahrbuch fi~r Kinderheilkunde. Hft. 1.
13. British reed. Journal. 1896. Nr. 1831.
14. Allgemeine u. sTecielle Theraloie fi~r J~inder~ranBheiten. 1896. (Russisch.)
15..Boat. reed. and surg. Journal. 3. Dec. 1896. Vol. CXXXV. - - Ref. H~/.qie,.
Rundsehau. 1897. Nr. 19.
16. Deutsche Vierteljah~'ssehrlft fi~r 5ffeutl. Gesundheitsfiflege. 1897. Bd. XXIX.
17. Berliner klin. ~Fochenachrift. 1897. Nr. 35--38.
18. Semaine mddicale. 1895. Nr. 68.
19. Diese Zeitsehrift. Bd. X X u
20. The Zancet. 23. Oct. 1897. - - Ref. Centralblatt fiPr Bakteriol. Bd. XXIV.
Nr. 10.
21. Semaine mddieale. 1897. Nr. 4 6 . .
22. l~evue de mdddcine. Janvier 1898. -- Ref. Mi~nchener mecl. Woehen~chrift.
1898. Nr. 15.
23 Deutsche reed. Wochenschrlft. 1898. Nr. 36.
2 4 . . E b e n d a . 1898. Nr. 6.
25. _Kinderheilkunde. 1898. Nr. 1. (Russisch.)
26. Brit. reed, Journ. 1898, Nr. 1946. - - .t~ygleu. Rundsehau. 1898. Nr. 19.
2 7 . . D i e s e Zcitschrift. 1898. Bd. XXIX.
28. Ergebnisse get allgera. _Pathologle. 1897. ffahrg. IV.
29. Diese Zeitschrift. 1899. Bd. XXXI.
30. Die Epidemiologie. W e y l ' s •andbueh dee tT,#giene. 1899. Bd. XXXVII.
70
G. G~BRITSCREWSKY: ZuR PROPRYLAXE I)ER DIPHTHERIE.
3I.
32.
Nr. 7.
33.
34.
35.
Revue d'hyglg~ne e~ de 2ooliee. sanitaire. 1899. T. XXI. ~r. ~.
British reed. Journal. 15. April 1899. - - Ref. ~.qien. RundseT~au. 1900.
2Vo~,k Magozin for Zazgevidensk~ben. 1900. Nr. 2.
~dinbour#h reed. Journal. Juni 1900.
~lrehi~e, ~'uase8 de Tat~ologie. 1899. T. VII. - - Au~;orreferat Cen~ralSla#~ f.
Bakteriologie. Bd. XXVI. Nr. 17/17.
36. Wratseh. 1900. Nr. 14. (Russisch.)
37. Vorlesungen i~ber aeute lnfeetionskrankheiten der .~inder. 1899. (Russisch.)
38. WraIaeh. 1899. (Russisch.) - - Ce~tralblatt fi~r J~aJcteriologie. Bd. XXVI.
Nr. 16/17,
|
|
https://openalex.org/W2886027067
|
https://europepmc.org/articles/pmc6084991?pdf=render
|
English
| null |
Estimated global overweight and obesity burden in pregnant women based on panel data model
|
PloS one
| 2,018
|
cc-by
| 8,535
|
RESEARCH ARTICLE OPEN ACCESS To estimate the global and country-level burden of overweight and obesity among pregnant
women from 2005 to 2014. Citation: Chen C, Xu X, Yan Y (2018) Estimated
global overweight and obesity burden in pregnant
women based on panel data model. PLoS ONE 13
(8): e0202183. https://doi.org/10.1371/journal. pone.0202183 Editor: Rebecca Painter, Amsterdam UMC, location
AMC, NETHERLANDS Editor: Rebecca Painter, Amsterdam UMC, location
AMC, NETHERLANDS Received: November 27, 2017
Accepted: July 29, 2018
Published: August 9, 2018 Copyright: © 2018 Chen et al. This is an open
access article distributed under the terms of the
Creative Commons Attribution License, which
permits unrestricted use, distribution, and
reproduction in any medium, provided the original
author and source are credited. Cheng Chen1, Xianglong Xu2,3,4, Yan Yan1* 1 Department of Epidemiology and Health Statistics, Xiangya School of Public Health, Central South
University, Changsha, Hunan, China, 2 School of Public Health and Management, Chongqing Medical
University, Chongqing, China, 3 Research Center for Medicine and Social Development, Chongqing Medical
University, Chongqing, China, 4 Collaborative Innovation Center of Social Risks Governance in Health,
Chongqing Medical University, Chongqing, China a1111111111
a1111111111
a1111111111
a1111111111
a1111111111 * 849352541@qq.com Methods Publicly accessible country-level data were collected from the World Health Organization,
the World Bank and the Food and Agricultural Organization. We estimated the number of
overweight and obese pregnant women among 184 countries and determined the time-
related trend from 2005 to 2014. Based on panel data model, we determined the effects of
food energy supply, urbanization, gross national income and female employment on the
number of overweight and obese pregnant women. Estimated global overweight and obesity
burden in pregnant women based on panel
data model Cheng Chen1, Xianglong Xu2,3,4, Yan Yan1* Results We estimated that 38.9 million overweight and obese pregnant women and 14.6 million
obese pregnant women existed globally in 2014. In upper middle income countries and
lower middle income countries, there were sharp increases in the number of overweight and
obese pregnant women. In 2014, the percentage of female with overweight and obesity in
India was 21.7%, and India had the largest number of overweight and obese pregnant
women (4.3 million), which accounted for 11.1% in the world. In the United States of
America, a third of women were obese, and the number of obese pregnant women was
1.1 million. In high income countries, caloric supply and urbanization were positively associ-
ated with the number of overweight and obese pregnant women. The percentage of employ-
ment in agriculture was inversely associated with the number of overweight and obese
pregnant women, but only in upper middle income countries and lower middle income
countries. Data Availability Statement: All relevant data are
within the paper and its Supporting Information
files. Introduction Obesity is a growing public health hazard worldwide. The proportion of global adult women
with overweight increased from 29.8% (29.3–30.2%) in 1980 to 38.0% (37.5–38.5%) in 2013,
and the increasing trend was observed in both high income and middle income countries [1]. Among pregnant women, increased body mass index (BMI) was associated with numerous
pregnancy related complications, including gestational diabetes mellitus (GDM), pregnancy
hypertension and preeclampsia [2, 3]. Women with overweight or obesity involved a relatively
high risk of severe maternal morbidity and mortality. Previous experts reported a odds ratio
(OR) for severe maternal morbidity of 1.1 for women with obesity class 1 (BMI 30.0–34.9)
compared with women with normal weight (BMI 18.5–24.9) [4]. The OR for obesity class 2
(BMI 35.0–39.8) was 1.2, and for obesity class 3 (BMI 40) was 1.4. Maternal obesity also
increased perinatal mortality. A previous cohort study found that maternal obesity was associ-
ated with nearly 25% of stillbirth that occurred between 37 and 42 weeks’ gestation [5]. In
addition, overweight and obesity were associated with elevated risks of fetal macrosomia, some
birth defects, and metabolic disease of children [6,7]. Considering the affect of pregnancy overweight and obesity on mothers and infants, it is
need to investigate the burden of overweight and obesity among pregnant women. Thirty-two
percent of Swedish pregnant women were overweight or obese in 2008–2010 [8]. The preva-
lence of overweight among pregnant women in Iceland increased form 25.9% to 27.7% within
nine years [9]. According to a retrospective cohort study in Canada, twenty-two percent of
pregnant women were obese and 24% were overweight in 2004–2014 [10]. A recent meta anal-
ysis reported that the prevalence of maternal obesity in Africa ranged from 6.5% to 50.7% [11]. However, previous studies exploring the prevalence of overweight and obesity among preg-
nant women were limited by the focus on a single country. The global burden of overweight
and obesity among pregnant women remained unclear. Many previous studies explored obesity of pregnant women and its determinants. For low-
income women, fast food intake can increase caloric supply, which is sufficient to explain the
increase of BMI in pregnant women [12, 13]. Compared with metropolitan residents, rural res-
idents were more likely to be overweight and obese in many countries, such as the United
States of America and China [14, 15]. Data Availability Statement: All relevant data are
within the paper and its Supporting Information
files. Funding: The funders had no role in study design,
data collection and analysis, decision to publish, or
preparation of the manuscript. The study was
supported by the National Natural Science
Foundation of China (81373101, 81673276). Competing interests: The authors have declared
that no competing interests exist. 1 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 Global overweight and obese pregnant women Conclusion The number of overweight and obese pregnant women has increased in high income and
middle income countries. Environmental changes could lead to increased caloric supply and
decreased energy expenditure among women. National and local governments should work
together to create a healthy food environment. PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 Global overweight and obese pregnant women group had a higher risk of obesity compared to those in the agricultural occupation group if
they had no education [18]. Although the World Health Organization (WHO) and the Global Burden of Disease study
(GBD) provided data of obesity and overweight data among adults [19], those data has not
been fully used to explore the prevalence of obesity and overweight among pregnant women. It is needed to explore more evidence about overweight and obesity among pregnant women. Therefore, the objectives of this study were to (1) estimate the global and country-level number
of overweight and obesity among pregnant women from 2005 to 2014; (2) identify relative
contributions of economic development, caloric supply, urbanization and female employment
to the number of overweight and obese pregnant women. Data sources We derived an estimate of the number of overweight and obese pregnant women using pub-
licly accessible country-level estimates of the following parameters: total population [20],
crude birth rate [21], estimated prevalence of overweight and obesity in female [22]. In each
country, the estimated overweight and obesity prevalence rate in female (>18 years) was age-
standardized. We collected overweight and obesity data of 195 countries, birth rate data of 255
countries and population data of 265 countries. We excluded countries with missing data, and
data of 184 countries form 2005 to 2014 were used in the final study. Eleven countries with
missing data were excluded, namely Cook Islands, Monaco, Nauru, Niue, Saint kitts and
Nevis, San Marino, South Sudan, Sudan, Sudan (former), Tuvalu and Dominica. To evaluate the contribution of energy intake to overweight and obesity, we collected data
of the food balance sheets (FBS) from the Food and Agricultural Organization (FAO) [23]. The
FBS data were compiled from national accounts of the supply and use of foods. The data pro-
vided a comprehensive picture of food consumption at country-level, and reflected the
increasing trend of per capita caloric supply. The database of FBS were updated in 2017, and
the latest data were food supply in 2013. To reflect the changes of social demographic and eco-
nomic characteristics, we also collected urbanization data, GNI data, and employment data
from the World Bank [24]. Urbanization was the percentage of population residing in urban
areas in each country according to national definition. GNI per capita data were in current U. S. dollars, divided by the midyear population, and deflated base on consumer price indexes. The indicators of employment were the percentages of employment in different industries of
all female employment, including employment in industries, employment in services and
employment in agriculture. Those factors were most ubiquitous in country-level and associ-
ated with the energy balance. In July 2017, we collected data of 184 countries form 2005 to
2013. PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 Introduction However, countries with high rates of urbanization usu-
ally have higher rates of obesity than those with low rates of urbanization. A previous system
review found a consistent positive association between urbanization and obesity in many
countries in Southeast Asia, and the association was greater in low gross national income
(GNI) countries [16]. Experts argued that urbanization could tip the balance between energy
intake and energy expenditure, namely decreases in physical activity and increases in the con-
sumption of cheap fast food [17]. Urbanization is usually accompanied by the transformation
of industrial structure. In middle income and low income countries, a growing number of
female work in service sectors. Occupational physical activity is an important determinant of
daily energy expenditure. A previous study found that women in the sedentary occupation PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 2 / 14 Estimating the burden of overweight and obesity in pregnant women BMI is defined as the weight in Kilograms divided by the square of the height in meters
(Kg/m2) [25]. According to data in the WHO, a BMI of 25.0 kg/m2 or more is classified as
overweight and obesity, and a BMI of 30.0 kg/m2 or more is defined as obesity. The point esti-
mated number of overweight pregnant women was obtained using the following formula:
Estimated number of overweight pregnant women = Total population×crude birth rate
280
365 estimated prevalence of overweight in female. We multiplied the total population by the crude birth rate, and then by the average gesta-
tional period (280 days), to calculate the number of pregnant days, per country. By dividing PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 3 / 14 Global overweight and obese pregnant women the number by 365 days, we estimated the number of women pregnant on any given days dur-
ing the year. Finally, by multiplying this number by the overweight prevalence, we calculated a
point estimate of the number of overweight and obese pregnant women. Similarly, the number
of obese pregnant women was calculated using the same method. This formula was adapted
form a previous study which provided a useful method to estimate the number of pregnant
women [26]. Country-level pointed estimates were added together to generate the global esti-
mates of the number of overweight and obese pregnant women. According to the 95% confi-
dence intervals of the overweight and obesity data, sensitivity analyses were used to provided
the upper and lower bounds of the estimate number of overweight and obese pregnant
women. Data analysis Panel data were often termed time series and cross section data[27]. Compared with singular
time series or cross-sectional analysis, panel data carried more information about the hetero-
geneity of individuals. The general model of the panel data can be described as the following
formula: yit ¼ ait þ mit þ bitxit yit refers to an explained variable and xit is an explanatory variable. i = 1. . .N refers to the indi-
vidual index. t = 1. . .T refers to the time index. αit is the intercept and μit shows the error term
with classic assumptions. βit represents the coefficient of xit. According to different interceptions, panel data model includes three kinds of model,
namely random effects model, pooled effects model and fixed effects model. We used the F test
to choose fixed or pooled effects specification. Then, we used the Hausman test to choose fixed
or random effects specification[28]. We used multivariable panel data models, and adjusted
beta coefficients were provided. Significance level was set as p <0.05, and the p value used a
two sided test. Microsoft Excel and R software version 3.3.1. were used to analyse these data. Results Countries were divided into four groups by the World Bank, namely high income countries
(HICs), upper middle income countries (UMICs), lower middle income countries (LMICs)
and low income countries (LICs). There were 52 HICs, 53 UMICs, 49 LMICs, and 30 LICs. We estimated that there were 38.9 million overweight and obese pregnant women and 14.6
million obese pregnant women in 2014 (Table 1). LMICs carried the greatest burden of over-
weight and obesity in pregnant women, and UMICs carried the greatest burden of obesity in
pregnant women. The burden of obesity in pregnant women was lower in LICs than in other
countries. Data of 184 countries was provided in S1 and S2 Figs. Table 1. Total number of overweight and obese pregnant women and percentage of global burden by WHO region in 2014. Income group
Number of overweight and obese pregnant women (BMI25)
Number of obese pregnant women (BMI>30)
Total
Percentage of global burden
Total
Percentage of global burden
High income
5275800
13.5%
2552100
17.5%
Upper middle income
13646600
35.0%
5507100
37.7%
Lower middle income
15237800
39.1%
5116400
35.0%
Low income
4786800
12.3%
1425400
9.8%
All countries combined
38947100
.. 14601100
.. https://doi org/10 1371/journal pone 0202183 t001 e 1. Total number of overweight and obese pregnant women and percentage of global burden by WHO region in 2014. Table 1. Total number of overweight and obese pregnant women and percentage of global burde PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 4 / 14 Global overweight and obese pregnant women Fig 1. Number of overweight and obese (BMI>25) pregnant women by WHO region from 2005 to 2014. h
//d i
/10 13 1/j
l
0202183 001 Fig 1. Number of overweight and obese (BMI>25) pregnant women by WHO region from 2005 to 2014. https://doi.org/10.1371/journal.pone.0202183.g001 https://doi.org/10.1371/journal.pone.0202183.g001 The increasing trends of overweight and obese pregnant women were observed in all
income groups, but with different increasing patterns (Figs 1 and 2). LICs had the lowest num-
ber of overweight and obese pregnant women for many years. The number of overweight and
obese pregnant women in UMICs and LMICs had a sharp increase. The number of obese Fig 2. Number of obese pregnant women (BMI30) by WHO region from 2005 to 2014. https://doi.org/10.1371/journal.pone.0202183.g002
PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018
5 / 14 Fig 2. Results Number of obese pregnant women (BMI30) by WHO region from 2005 to 2014. https://doi.org/10.1371/journal.pone.0202183.g002 PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 5 / 14 Global overweight and obese pregnant women pregnant women in UMICs was at a high level from 2005 to 2013. Although the number of
obese pregnant women in LICs was at a low level, there was a increasing trend over that time
period. pregnant women in UMICs was at a high level from 2005 to 2013. Although the number of
obese pregnant women in LICs was at a low level, there was a increasing trend over that time
period. Estimates for 20 countries with the highest overweight and obesity burden in pregnant
women were presented in Tables 2 and 3. In 2014, the percentage of female with overweight
and obesity in India was 21.7%. India had the largest number of overweight and obese preg-
nant women (4.3 million), which accounted for the largest proportion (11.1%) in the world. The increases of overweight and obese pregnant women in some countries were more than
50%, such as Nigeria (55.4%), Democratic Republic of the Congo (53.4%) and United Republic
of Tanzania (59.3%). For some countries with a high rate of overweight and obesity, the
changes in ten years were small, such as United States of America (3.8%), Mexico (5.3%) and
Turkey (5.9%). As the birth rate of Brazil decreased form 18.078 per 1000 people in 2005 to
14.727 per 1000 people in 2014, the number of overweight and obese pregnant women
decreased by 1.7%. The United States of America had the largest number of obese pregnant
women (1.07 million) in 2014. China also had 1.06 million obese pregnant women, and the
number increased by 71.2% in ten years. For some countries with a high birth rate, the number
of obese pregnant women was even doubled in ten years, such as Nigeria (96.9%), Democratic
Republic of the Congo (102.2%), and United Republic of Tanzania (111.6%). The changes in urbanization of different income groups were presented in S1 Table. In
2013, the urbanization rate in HICs and UMICs reached 80.6% and 63.7%, respectively. The
urbanization rate in LMICs increased form 38.5% in 2005 to 41.5% in 2013. GNI of UMICs
and LMICs increased by 50.2% and 54.0%, respectively (S2 Table). The Changes in caloric sup-
ply were presented in S3 Table. Results Caloric supply in HICs increased from 3221.0 kcal/capita/day
in 2005 to 3263 kcal/capita/day in 2013. Caloric supply in LICs was 2324.4 kcal/capita/day in
2013, and increased by 5.8% (128 kcal/capita/day) in nine years. For many countries, the per-
centage of employment in agriculture decreased, while the percentage of employment in ser-
vices increased (S4 Table). In 2013, the percentage of employment in agriculture was 1.8% in
HICs, 9.3% in UMICs, 37.4% in LMICs, and 73.6% in LICs. For female in LMICs, the percent-
age of employment in services increased form 39.5% in 2005 to 50.7% in 2013. As three indicators of employment were related to each other, we chose the percentage of
employment in agriculture as the proxy of changes in occupational physical activity. Accord-
ing to the results of F test and Hausman test, random effects model was used for HICs, UMICs
and LICs, and fixed effects model was used for LIMCs. For HICs, caloric supply (p = 0.001)
and urbanization (p = 0.026) were positively associated with the number of overweight and
obese pregnant women, and GNI (p = 0.004) was significantly associated with the number of
obese pregnant women (Table 4). For UMICs and LMICs, the effect of caloric supply on the
number of overweight and obese pregnant women was insignificant, and the percentage of
employment in agriculture was inverse associated with the number of overweight and obese
pregnant women. For LICs, urbanization (p = 0.005) and GNI (p<0.001)were significantly
associated with the number of overweight and obese pregnant women. Discussion The large number of overweight and obese pregnant women was a huge burden on health
care. This study estimated that nearly forty million pregnant women were overweight or obese
in the world in 2014. More than 70% of overweight pregnant women occurred in UMICs and
LMICs, owing to a large population and a high birth rate in those countries. The number of
overweight and obese pregnant women increased rapidly in middle income countries from
2005 to 2014, especially in India, China and Nigeria. In many countries, more than half of 6 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 women were overweight, and nearly a third of women were obese, such as Egypt, Turkey, Iran,
and South Africa. More adverse maternal and fetal outcomes were observed in women with
overweight and obesity. A previous study in Iranian found that pregnant women with obesity Global overweight and obese pregnant women Table 2. Total number of overweight and obese (BMI>25kg/m2) pregnant women and rate of overweight among female for the 20 high overweight burden
countries. Country
Rate of overweight and
obese among female [95%
CI ]
Number of overweight and obese pregnant
women [95% CI]
Changes in 10
years
Percentage of global burden
in 2014
2005
2014
2005
2014
Indiac
16.6 [14.4–
19.0]
21.7 [17.5–
26.4]
3518500 [3090400–
3997600]
4302000 [3509000–
5180300]
22.3%
11.1%
Chinab
25.8 [22.9–
28.8]
33.1 [27.4–
39.2]
3199500 [2862200–
3556700]
4285100 [3591200–
5038500]
33.9%
11.0%
Nigeriac
30.3 [26.7–
34.1]
39.6 [33.9–
45.3]
1373600 [1217600–
1539500]
2134900 [1845400–
2431500]
55.4%
5.5%
United States of Americaa
58.4 [55.2–
61.7]
62.9 [57.9–
67.8]
1853400 [1756600–
1952500]
1923400 [1780900–
2065600]
3.8%
4.9%
Egyptc
64.2 [60.4–
68.1]
70.2 [64.6–
75.3]
921100 [869500–
973800]
1340900 [1239500–
1434500]
45.6%
3.5%
Brazilb
48.0 [44.1–
51.9]
53.0 [46.9–
59.3]
1254600 [1157400–
1351000]
1233900 [1099800–
1375200]
-1.7%
3.2%
Mexicob
60.8 [57.0–
64.6]
65.2 [59.3–
70.7]
1119400 [1053500–
1185700]
1178300 [1076200–
1273400]
5.3%
3.0%
Indonesiac
22.1 [18.8–
25.6]
28.3 [22.9–
34.2]
823700 [705200–
947500]
1102300 [907500–
1318900]
33.8%
2.8%
Pakistanc
21.0 [17.4–
25.2]
26.0 [20.4–
32.0]
741700 [621200–
878000]
1079400 [859300–
1315200]
45.5%
2.8%
Russian Federationb
53.5 [49.5–
57.5]
54.8 [47.7–
61.5]
600700 [558000–
644200]
804100 [707500–
897800]
33.8%
2.1%
Turkeyb
63.5 [60.1–
66.8]
68.4 [63.4–
73.2]
645100 [612400–
676800]
682900 [635500–
727900]
5.9%
1.8%
Islamic Republic of Iranb
58.1 [55.0–
61.2]
64.2 [58.7–
69.4]
554800 [526600–
583200]
675100 [619800–
726000]
21.7%
1.7%
Democratic Republic of the
Congod
22.1 [17.5–
27.3]
27.7 [21.0–
35.2]
435200 [349300–
533300]
667600 [514800–
840900]
53.4%
1.7%
Iraqb
58.4 [53.4–
63.2]
63.7 [57.1–
70.0]
425200 [390400–
458300]
596200 [537600–
653600]
40.2%
1.5%
Ethiopiad
18.5 [15.0–
22.3]
24.2 [18.5–
30.2]
421700 [345100–
503300]
583400 [452700–
718600]
38.3%
1.5%
South Africab
58.6 [54.4–
62.9]
64.1 [58.5–
69.4]
492800 [459100–
527100]
550000 [504000–
593300]
11.6%
1.4%
United Republic of
Tanzaniad
24.7 [21.1–
28.5]
32.1 [26.6–
37.9]
312500 [268900–
358000]
497800 [417500–
583800]
59.3%
1.3%
Bangladeshc
15.0 [12.2–
18.1]
20.4 [15.8–
25.8]
397200 [328600–
474200]
492800 [387900–
615000]
24.1%
1.3%
Philippinesc
22.2 [18.9–
25.7]
26.9 [21.3–
32.9]
399500 [343700–
458600]
481800 [386300–
584200]
20.6%
1.2%
Algeriab
56.2 [51.5–
60.9]
61.3 [54.8–
67.4]
298600 [275000–
322500]
445000 [400700–
486900]
49.0%
1.1% overweight among female for the 20 high overweight burden 7 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 were 4 times more likely to develop gestational hypertension compared to those with normal
weight [29]. Maternal obesity also increased the risk of fetal macrosomia, cardiac breaks, neu-
ral tube defects, and fetal death [30, 31, 32]. Health care providers should pay more attention
to the adverse effects of obesity on maternal and fetal. Global overweight and obese pregnant women Table 3. Total number of obese pregnant women,rate of obesity among female for the 20 high obesity burden countries. Country
Rate of obesity among
female [95% CI]
Number of obese pregnant women [95%
CI]
Changes in 10
years
Percentage of global
burden in 2014
2005
2014
2005
2014
United States of Americaa
30.2 [26.9–
33.7]
34.9 [29.6–
40.4]
958400 [860000–
1064700]
1067200 [914000–
1226200]
11.3%
7.3%
Chinab
5.0 [3.8–6.3]
8.2 [5.5–
11.7]
620000 [479900–
773800]
1061500 [731400–
1491300]
71.2%
7.3%
Indiac
3.2 [2.5–4.1]
5.1 [3.4–7.2]
678200 [534100–
862600]
1011000 [699800–
1407500]
49.1%
6.9%
Nigeriac
9.3 [7.1–
11.6]
15.4 [11.4–
19.9]
421600 [326800–
521300]
830200 [625900–
1059400]
96.9%
5.7%
Egyptc
33.3 [29.1–
37.7]
39.7 [33.2–
46.1]
477700 [420400–
537700]
758300 [640200–
873500]
58.7%
5.2%
Mexicob
27.5 [23.8–
31.5]
32.4 [26.4–
38.7]
506300 [442600–
575700]
585500 [483200–
693100]
15.7%
4.0%
Brazilb
19.2 [16.2–
22.3]
24.0 [19.1–
29.4]
501800 [427600–
577300]
558700 [450400–
679500]
11.3%
3.8%
Russian Federationb
24.5 [21.0–
28.3]
26.2 [20.1–
32.7]
275100 [237600–
316400]
384400 [298100–
475400]
39.7%
2.6%
Turkeyb
30.8 [27.3–
34.3]
36.2 [31.0–
41.8]
312900 [279100–
346800]
361400 [313100–
414700]
15.5%
2.5%
Pakistanc
5.7 [3.9–8.2]
8.2 [5.2–
12.0]
201300 [141200–
286400]
340400 [225000–
489800]
69.1%
2.3%
South Africab
32.7 [28.5–
37.0]
38.1 [32.6–
43.9]
182600 [133400–
247100]
315500 [213400–
449500]
18.9%
2.2%
Indonesiac
4.9 [3.5–6.7]
8.1 [5.3–
11.7]
275000 [241900–
309300]
326900 [281600–
373500]
72.7%
2.2%
Islamic Republic of Iranb
24.8 [21.9–
27.8]
29.7 [24.6–
35.3]
236800 [210400–
264100]
312300 [261700–
368400]
31.9%
2.1%
Iraqb
26.8 [21.2–
32.7]
32.1 [24.5–
39.9]
195100 [156400–
235600]
300400 [234200–
368000]
54.0%
2.1%
Algeriab
25.3 [20.3–
30.6]
29.7 [23.0–
37.0]
134400 [109300–
161400]
215600 [169000–
265200]
60.4%
1.5%
Saudi Arabiab
35.6 [31.2–
40.0]
40.7 [33.9–
47.3]
90500 [57100–
135100]
183100 [112300–
281000]
23.1%
1.3%
Democratic Republic of the Congod
4.6 [2.8–7.0]
7.6 [4.4–
11.9]
157700 [139500–
176300]
194200 [163700–
224200]
102.2%
1.3%
Argentinab
24.5 [19.9–
29.3]
30.1 [23.5–
37.1]
138500 [113500–
164100]
174100 [137900–
213200]
25.7%
1.2%
United Kingdom of Great Britain and
Northern Irelanda
23.4 [21.1–
25.8]
28.4 [24.3–
32.5]
130100 [117800–
142900]
168900 [145900–
192100]
29.8%
1.2%
United Republic of Tanzaniad
6.2 [4.4–8.4]
10.7 [7.4–
14.8]
78400 [56500–
105100]
165900 [118000–
227000]
111.6%
1.1%
Note. CI: confidence intervals were 4 times more likely to develop gestational hypertension compared to those with normal
weight [29]. Global overweight and obese pregnant women mber of overweight and obese pregnant women based on panel data model between 2005 and 2013. Table 4. Factors associated with the number of overweight and obese pregnant women based on panel data model between 2005 and 2013. Income group
Variable
BMI25
BMI>30
B
S.E. p-value
B
S.E. p-value
Hig income
Caloric supply a
26.66
8.22
0.001
4.69
4.93
0.342
Urbanization b
1270.40
567.42
0.026
1432.70
337.98
<0.001
Gross national income c
0.06
0.13
0.628
0.22
0.08
0.004
Employment in agriculture d
-281.23
635.75
0.658
91.69
381.02
0.810
Upper middle income
Caloric supply
56.79
38.32
0.139
28.71
16.28
0.079
Urbanization
8298.60
1736.20
<0.001
3606.40
704.35
<0.001
Gross national income
3.73
1.54
0.016
2.81
0.66
<0.001
Employment in agriculture
-2407.60
791.85
0.003
-1080.90
337.15
0.001
Lower middle income
Caloric supply
-5.98
56.58
0.916
-6.75
28.28
0.811
Urbanization
6055.86
2755.61
0.029
2528.21
1377.31
0.067
Gross national income
14.33
5.35
0.008
7.70
2.67
0.004
Employment in agriculture
-4558.11
915.53
<0.001
-2437.66
457.60
0.000
Low middle income
Caloric supply
-52.11
26.67
0.052
-16.59
10.58
0.118
Urbanization
3022.80
1074.76
0.005
1045.45
388.62
0.008
Gross national income
82.61
10.70
<0.001
35.98
4.28
<0.001
Employment in agriculture
665.17
451.05
0.142
267.44
178.75
0.136 For HICs, the burden of overweight and obesity among pregnant women has been in a high
level for many years.The increases of the number of overweight and obese pregnant women in
UMICs and LMICs were faster than those in HICs. Those changes suggested a worldwide
time-related phenomenon rather than a country-specific trend [33]. Previous studies found a
slowdown in the increase rate of overweight and obesity in HICs, which provided some hope
that the epidemic might had peaked in developed countries and that the populations in middle
income countries might not reach the very high rates of over 40% [1]. However, considering
the large population and the increasing rate of overweight in middle income countries, the
burden of maternal overweight in those countries would be more serious in future. Given that an increasing number of people lived in urban area, food environment and dis-
eases of urban residents changed a lot [33, 34]. We found that urbanization was associated
with the increasing number of overweight and obese pregnant women. City life can be more
sedentary than rural life. Factors associated with the number of overweight and obese pregnant women based on panel data model between 2005 and 2013. Income group
Variable
BMI25
BMI>30
B
S.E. p-value
B
S.E. p-value
Hig income
Caloric supply a
26.66
8.22
0.001
4.69
4.93
0.342
Urbanization b
1270.40
567.42
0.026
1432.70
337.98
<0.001
Gross national income c
0.06
0.13
0.628
0.22
0.08
0.004
Employment in agriculture d
-281.23
635.75
0.658
91.69
381.02
0.810
Upper middle income
Caloric supply
56.79
38.32
0.139
28.71
16.28
0.079
Urbanization
8298.60
1736.20
<0.001
3606.40
704.35
<0.001
Gross national income
3.73
1.54
0.016
2.81
0.66
<0.001
Employment in agriculture
-2407.60
791.85
0.003
-1080.90
337.15
0.001
Lower middle income
Caloric supply
-5.98
56.58
0.916
-6.75
28.28
0.811
Urbanization
6055.86
2755.61
0.029
2528.21
1377.31
0.067
Gross national income
14.33
5.35
0.008
7.70
2.67
0.004
Employment in agriculture
-4558.11
915.53
<0.001
-2437.66
457.60
0.000
Low middle income
Caloric supply
-52.11
26.67
0.052
-16.59
10.58
0.118
Urbanization
3022.80
1074.76
0.005
1045.45
388.62
0.008
Gross national income
82.61
10.70
<0.001
35.98
4.28
<0.001
Employment in agriculture
665.17
451.05
0.142
267.44
178.75
0.136
Note. a kcal/capita/day
b % of total population
c current US$
d % of female employment. https://doi.org/10.1371/journal.pone.0202183.t004
Global overweight and obese pregnant women
PLOS ONE | htt
//d i
/10 1371/j
l
0202183
A
t 9 2018
9 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 Maternal obesity also increased the risk of fetal macrosomia, cardiac breaks, neu-
ral tube defects, and fetal death [30, 31, 32]. Health care providers should pay more attention
to the adverse effects of obesity on maternal and fetal. 8 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 For HICs, the burden of overweight and obesity among pregnant women has been in a high
level for many years.The increases of the number of overweight and obese pregnant women in
UMICs and LMICs were faster than those in HICs. Those changes suggested a worldwide
time-related phenomenon rather than a country-specific trend [33]. Previous studies found a
slowdown in the increase rate of overweight and obesity in HICs, which provided some hope
that the epidemic might had peaked in developed countries and that the populations in middle
income countries might not reach the very high rates of over 40% [1]. However, considering
the large population and the increasing rate of overweight in middle income countries, the
burden of maternal overweight in those countries would be more serious in future. Given that an increasing number of people lived in urban area, food environment and dis-
eases of urban residents changed a lot [33, 34]. We found that urbanization was associated
with the increasing number of overweight and obese pregnant women. City life can be more
sedentary than rural life. A previous study found that BMI of urban residents was lower in
countries with more land devoted to parks, which were sites for physical activity,walking and
cycling [35]. A recent study in Seoul found that the number of sports facilities in urban were
negatively associated with the probability of obesity [36]. City life also changes the availability
of food, especially fast foods and energy-dense foods. Previous studies found that supermarkets
were associated with a higher BMI among black adults [37]. The presence of convenience
stores and fast food restaurants was a driver of weight excess, which usually offered energy-
dense foods [38]. Although similar results shows that city life is associated with a higher risk of
obesity than rural life, findings in the literature are not always consistent. A previous study
found that the prevalence of obesity among women was higher in rural than in urban (33.4%
Table 4. https://doi.org/10.1371/journal.pone.0202183.t004 A previous study found that BMI of urban residents was lower in
countries with more land devoted to parks, which were sites for physical activity,walking and
cycling [35]. A recent study in Seoul found that the number of sports facilities in urban were
negatively associated with the probability of obesity [36]. City life also changes the availability
of food, especially fast foods and energy-dense foods. Previous studies found that supermarkets
were associated with a higher BMI among black adults [37]. The presence of convenience
stores and fast food restaurants was a driver of weight excess, which usually offered energy-
dense foods [38]. Although similar results shows that city life is associated with a higher risk of
obesity than rural life, findings in the literature are not always consistent. A previous study
found that the prevalence of obesity among women was higher in rural than in urban (33.4% PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 9 / 14 Global overweight and obese pregnant women vs 28.2%), and potential risk factors were lower leisure-time, intake of fiber and fruits and
higher intake of sweetened beverages [39]. We found that food energy supply increased in many countries from 2005 to 2013. Previous
studies in Venezuela and Ireland also reported a increasing trend of energy supply between
1961 and 2007 [40, 41]. We found that energy supply in HICs has been in a high level for
many years. Compared with China and Japan, the consumption of total meat was higher in
European Union, the United State of America and Canada [42]. This study found that caloric
supply was a risk factor for the huge number of pregnant women with overweight in HICs, but
not in other income group countries. A previous study about 69 countries also reported that
the association between the change in food energy supply and the change in average body
weight was significant for HICs [43]. For LICs, the increase of caloric supply might be a sign of
improved nutrition. This study found that GNI was positively associated with the number of obese pregnant
women in all income groups. A previous study in thirty-three less developed countries found
that GNI was positively associated with overweight among mothers [44]. Economic develop-
ment can reduce food prices, especially prices of unhealthful foods. A previous study even
reported that approximately 18% of growth in obesity could be attributed to relative food
prices reduction between 1976 and 2001 [45]. This study found that the percentage of employ-
ment in agriculture was inversely associated with the number of overweight and obese preg-
nant women, but only in UMICs and LMICs. The main change in UMICs and LMICs was that
a growing number of women were occupied in service sectors rather than in agriculture. Owing to the reduction in occupational physical activity, daily energy output among women
has decreased by more than 100 kcal/day over the past 5 decades [46]. A study in Malaysia also
reported that low occupational physical activity in middle-aged women was associated with
higher risks of obesity and abdominal obesity [47]. Considering a growing number of overweight and obese pregnant women in both high
income and middle income countries, health workers are faced with a huge challenge of reduc-
ing unfavorable pregnancy outcomes. According to the Institute of Medicine, the recommend
GWG for overweight pregnant women is 7–11.9 kg and for obese pregnant women is 5–9 kg
[48]. Dietary interventions and physical activity interventions were recommended to limit
GWG and prevent GDM in overweight and obese pregnant women [49, 50, 51]. However, a
randomised controlled trail in UK found that dietary and physical interventions in pregnant
women with obesity were not adequate to prevent GDM or large-for-gestational-age infants,
and a recent study in Australia also reported no significant differences in GDM between the
behavioural nutrition intervention group and the control group after adjusting confounding
factors [52,53]. From a public health perspective, it is a cost-effective strategy to control the
prevalence of obesity among women of childbearing age. Women should be informed the
potential risk of fast food and the importance of a normal weight for pregnant women. As the
environment makes it easier to become overweight and obese, national and local governments
should promote a health food environment, such as portion control, high calories food avail-
ability and media restrictions [54]. Some limitations exist in this study. Firstly, the data of overweight and obesity on reproduc-
tive age might be better than those across the whole age range. Unfortunately, data on repro-
ductive age of many countries were not available form public accessible database. PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 Conclusion There was a great increase of the number of overweight and obese pregnant women in both
high income and middle income countries. Those data demonstrated that food energy supply,
urbanization rate, GNI and employment in agriculture were associated with the burden of
overweight and obese among pregnant women. In order to control obesity among pregnant
women, national and local governments need to create a healthy food environment. As the status
of overweight and obesity can last for a long time, the present data can be used to approximate
the number of overweight and obese pregnant women. Secondly, the definition of overweight
and obesity is different in different regions, which can not be reflected in these international
data. Overweight is defined as a BMI 25.0 to <30.0 kg/m2 by the WHO, and a BMI of 30.0 kg/
m2 or more is defined as obesity. However, WHO Asia Pacific guidelines suggest that PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 10 / 14 Global overweight and obese pregnant women overweight is defined as BMI 23–27.49 kg/m2, and obesity is defined as BMI 27.5kg/m2 [55]. The overweight and obesity rate in some Asia countries would be underestimated using the
former definition [55]. In the 2011 China Health and Nutrition Survey, obesity was defined as
BMI 28.0 kg/m2, and the age-adjusted prevalence of obesity among women was 11.0%,
which was higher than the prevalence provided by the WHO (7.1%) [56]. Thirdly, the level of
urbanization, namely large metropolitan, small metropolitan and micropolitan, is also an
important factor. Urbanization rate can not reflect these important information. Finally, this
study used country as the unit of analysis in the panel data model, which might lead to ecologi-
cal fallacy. We should not use country-level statistical findings to make inferences about the
energy balance of individuals. Author Contributions Formal analysis: Cheng Chen. Supervision: Yan Yan. Writing – original draft: Cheng Chen, Xianglong Xu. Writing – original draft: Cheng Chen, Xianglong Xu. S3 Table. Changes of food supply in different income groups from 2005 to 2013.
(DOC) S3 Table. Changes of food supply in different income groups from 2005 to 2013. (DOC) S3 Table. Changes of food supply in different income groups from 2005 to 2013. (DOC) S4 Table. Employment in different industries of all female employment from 2005 to
2013. (DOC) S1 Fig. The estimated number of overweight and obese pregnant women in 184 countries
in 2014.
(TIF) S1 Fig. The estimated number of overweight and obese pregnant women in 184 countries
in 2014. (TIF) Supporting information pp
g
S1 Fig. The estimated number of overweight and obese pregnant women in 184 countries
in 2014. (TIF)
S2 Fig. The estimated number of obese pregnant women in 184 countries in 2014. (TIF)
S1 Table. Changes of urban population in different income groups from 2005 to 2013. (DOC)
S2 Table. Changes of gross national income in different income groups from 2005 to 2013. (DOC)
S3 Table. Changes of food supply in different income groups from 2005 to 2013. (DOC)
S4 Table. Employment in different industries of all female employment from 2005 to
2013. (DOC)
Author Contributions
Formal analysis: Cheng Chen. Supervision: Yan Yan. Writing – original draft: Cheng Chen, Xianglong Xu. References
1. Ng M, Fleming T, Robinson M, Thomson B, Graetz N, Margono C, et al. Global, regional, and national
prevalence of overweight and obesity in children and adults during 1980–2013: a systematic analysis
for the Global Burden of Disease Study 2013. The Lancet. 2014; 384 (9945):766–781. https://doi.org/
10.1016/S0140-6736(14)60460-8 PMID: 24880830 pp
g
S1 Fig. The estimated number of overweight and obese pregnant women in 184 countries
in 2014. (TIF)
S2 Fig. The estimated number of obese pregnant women in 184 countries in 2014. (TIF)
S1 Table. Changes of urban population in different income groups from 2005 to 2013. (DOC)
S2 Table. Changes of gross national income in different income groups from 2005 to 2013. (DOC)
S3 Table. Changes of food supply in different income groups from 2005 to 2013. (DOC)
S4 Table. Employment in different industries of all female employment from 2005 to
2013. (DOC) S1 Table. Changes of urban population in different income groups from 2005 to 2013.
(DOC) S2 Table. Changes of gross national income in different income groups from 2005 to 2013. (DOC) S2 Table. Changes of gross national income in different income groups from 2005 to 2013. (DOC) References 1. Ng M, Fleming T, Robinson M, Thomson B, Graetz N, Margono C, et al. Global, regional, and national
prevalence of overweight and obesity in children and adults during 1980–2013: a systematic analysis
for the Global Burden of Disease Study 2013. The Lancet. 2014; 384 (9945):766–781. https://doi.org/
10.1016/S0140-6736(14)60460-8 PMID: 24880830 1. Ng M, Fleming T, Robinson M, Thomson B, Graetz N, Margono C, et al. Global, regional, and national
prevalence of overweight and obesity in children and adults during 1980–2013: a systematic analysis
for the Global Burden of Disease Study 2013. The Lancet. 2014; 384 (9945):766–781. https://doi.org/
10.1016/S0140-6736(14)60460-8 PMID: 24880830 PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 11 / 14 Global overweight and obese pregnant women 2. Marchi J, Berg M, Dencker A, Olander EK, Begley C. Risks associated with obesity in pregnancy, for
the mother and baby: a systematic review of reviews. Obesity Reviews. 2015; 16(8):621–638. https://
doi.org/10.1111/obr.12288 PMID: 26016557 3. Ijas H, Morin-Papunen L, Keranen AK, Bloigu R, Ruokonen A, Puukka K, et al. Pre-pregnancy over-
weight overtakes gestational diabetes as a risk factor for subsequent metabolic syndrome. Eur J Endo-
crinol. 2013; 169(5):605–611. https://doi.org/10.1530/EJE-13-0412 PMID: 23959786 4. Lisonkova S, Muraca GM, Potts J, Liauw J, Chan WS, Skoll A, et al. Association Between Prepreg-
nancy Body Mass Index and Severe Maternal Morbidity. JAMA. 2017; 318 (18):1777–1786. https://doi. org/10.1001/jama.2017.16191 PMID: 29136442 5. Yao R, Ananth CV, Park BY, Pereira L, Plante LA, Perinatal Research Consortium. Obesity and the risk
of stillbirth: a population-based cohort study. Am J Obstet Gynecol. 2014; 210(5):451–457. https://doi. org/10.1016/j.ajog.2014.01.044 PMID: 24674712 6. Wang LF, Wang HJ, Ao D, Liu Z, Wang Y, Yang HX. Influence of pre-pregnancy obesity on the develop-
ment of macrosomia and large for gestational age in women with or without gestational diabetes mellitus
in Chinese population. J Perinatol. 2015; 35(12):985–990. https://doi.org/10.1038/jp.2015.119 PMID:
26401753 7. Parnell AS, Correa A, Reece EA. Pre-pregnancy Obesity as a Modifier of Gestational Diabetes and
Birth Defects Associations: A Systematic Review. Maternal and Child Health Journal 2017; 21
(5):1105–1120. https://doi.org/10.1007/s10995-016-2209-4 PMID: 28120287 8. Bjermo H, Lind S, Rasmussen F. The educational gradient of obesity increases among Swedish preg-
nant women: a register-based study. BMC Public Health 2015; 15:315. https://doi.org/10.1186/s12889-
015-1624-6 PMID: 25886465. 9. Eiriksdottir VH, Valdimarsdottir UA, Asgeirsdottir TL, Gisladottir A, Lund SH, Hauksdottir A, et al. Smok-
ing and obesity among pregnant women in Iceland 2001–2010. European Journal of Public Health
2015; 25(4):638–643. PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 References 23. Food and Agriculture Organization of the United Nations. Food Balance Sheets. [cited 2017 Jun 23]. Available from: http://www.fao.org/faostat/en/#data/FBS. 24. The World Bank. World Development Indicators. [cited 2017 Jun 23]. Available from: http://data. worldbank.org/indicator/NY.GNP.MKTP.PP.CD?view=chart. 25. World Health Organization, Obesity: preventing and managing the global epidemic. Report of a WHO
Consultation (WHO Technical Report Series 894). Geneva: World Health Organization, 2000. 26. Sugarman J, Colvin C, Moran AC, Oxlade O. Tuberculosis in pregnancy: an estimate of the global bur-
den of disease. Lancet Glob Health. 2014; 2(12):e710–6. https://doi.org/10.1016/S2214-109X(14)
70330-4 PMID: 25433626. 27. Deation A. Panel data model time series of cross-sections. Journal of Econometrics 1985; 30:109–26. 28. Sha T, Yan Y, Gao X, Xiang S, Zeng G, Liu S, et al. Association between Sleep and Body Weight: A
Panel Data Model Based on a Retrospective Longitudinal Cohort of Chinese Infants. Int J Environ Res
Public Health. 2017; 14(5). PMID: 28441347 29. Kazemian E, Sotoudeh G, Dorosty-Motlagh AR, Eshraghian MR, Bagheri M. Maternal obesity and
energy intake as risk factors of pregnancy-induced hypertension among Iranian women. J Health Popul
Nutr. 2014; 32(3):486–93. PMID: 25395911. 30. Talebian A, Soltani B, Sehat M, Zahedi A, Noorian A, Talebian M. Incidence and Risk Factors of Neural
Tube Defects in Kashan, Central Iran. Iran J Child Neurol. 2015; 9(3):50–6. PMID: 26401153. 31. Taheri M, Dehghani A, Noorishadkam M, Tabatabaei SM. Population attributable danger of hereditary
heart breaks. Risk factors among newborns in Yazd, Iran. J Med Life. 2015; 8(Spec Iss 3): 212–7. PMID: 28316693. 32. Usta A, Usta CS, Yildiz A, Ozcaglayan R, Dalkiran ES, et al. Frequency of fetal macrosomia and the
associated risk factors in pregnancies without gestational diabetes mellitus. Pan Afr Med J. 2017;
26:62. https://doi.org/10.11604/pamj.2017.26.62.11440 PMID: 28451039. 33. Wu Y, Xue H, Wang H, Su C, Du S, Wang Y. The impact of urbanization on the community food environ-
ment in China. Asia Pac J Clin Nutr. 2017; 26(3):504–13. https://doi.org/10.6133/apjcn.032016.09
PMID: 28429917. 34. Goryakin Y., Rocco L., Suhrcke M. The contribution of urbanization to non-communicable diseases:
Evidence from 173 countries from 1980 to 2008. Economics & Human Biology. 2017; 26:151–63. https://doi.org/10.1016/j.ehb.2017.03.004 PMID: 28410489. 35. Ewing R, Meakins G, Hamidi S, Nelson AC. Relationship between urban sprawl and physical activity,
obesity, and morbidity–Update and refinement. Health & Place. 2014; 26:118–26. https://doi.org/10. 1016/j.healthplace.2013.12.008 PMID: 24434082. 36. Kim J, Shon C, & Yi S. The Relationship between Obesity and Urban Environment in Seoul. References https://doi.org/10.1093/eurpub/ckv047 PMID: 25829507. 10. MacInnis N, Woolcott CG, McDonald S, Kuhle S. Population Attributable Risk Fractions of Maternal
Overweight and Obesity for Adverse Perinatal Outcomes. Scientific Reports. 2016; 6:22895. https://
doi.org/10.1038/srep22895 PMID: 26961675. 11. Onubi OJ, Marais D, Aucott L, Okonofua F, Poobalan AS. Maternal obesity in Africa: a systematic
review and meta-analysis. Journal of Public Health. 2015; 38(3):e218–e231. https://doi.org/10.1093/
pubmed/fdv138 PMID: 26487702. 12. Chang M, Brown R, Nitzke S. Fast Food Intake in Relation to Employment Status, Stress, Depression,
and Dietary Behaviors in Low-Income Overweight and Obese Pregnant Women. Maternal and Child
Health Journal. 2016; 20(7): 1506–17. https://doi.org/10.1007/s10995-016-1949-5 PMID: 26973147. 13. Fowles ER, Timmerman GM, Bryant M, Kim S. Eating at Fast-Food Restaurants and Dietary Quality in
Low-Income Pregnant Women. Western Journal of Nursing Research 2011; 33(5):630–651. https://
doi.org/10.1177/0193945910389083 PMID: 21131508. 14. Tian X, Zhao G, Li Y, Wang L, Shi Y. Overweight and obesity difference of Chinese population between
different urbanization levels. J Rural Health. 2014; 30(1):101–12. https://doi.org/10.1111/jrh.12041
PMID: 24383489. 15. Voss JD, Masuoka P, Webber BJ, Scher AI, Atkinson RL. Association of elevation, urbanization and
ambient temperature with obesity prevalence in the United States. Int J Obes (Lond). 2013; 37
(10):1407–12. https://doi.org/10.1038/ijo.2013.5 PMID: 23357956. 16. Angkurawaranon C, Jiraporncharoen W, Chenthanakij B, Doyle P, Nitsch D. Urban Environments and
Obesity in Southeast Asia: A Systematic Review, Meta-Analysis and Meta-Regression. PLoS ONE. 2014; 9(11):e113547. https://doi.org/10.1371/journal.pone.0113547 PMID: 25426942. 17. Bleich S, Cutler D FAU Murray C, Murray C FAU Adams A, Adams A. Why is the developed world
obese? Annual Review of Public Health. 2008; 29:273–95. https://doi.org/10.1146/annurev.publhealth. 29.020907.090954 PMID: 18173389. 18. Aitsi-Selmi A, Chen R, Shipley MJ, Marmot MG. Education is associated with lower levels of abdominal
obesity in women with a non-agricultural occupation: an interaction study using China’s Four Provinces
survey. BMC Public Health. 2013; 13:769. https://doi.org/10.1186/1471-2458-13-769 PMID:
23962144. 19. GBD 2015 obesity collaborators, Afshin A, Forouzanfar MH, Reitsma MB, Sur P, Estep K, et al. Health
Effects of Overweight and Obesity in 195 Countries over 25 Years. N Engl J Med. 2017; 377(1):13–27. https://doi.org/10.1056/NEJMoa1614362 PMID: 28604169. 20. United Nations. World Population Prospects 2017. [cited 2017 Jun 23]. Available from: https://esa.un. org/unpd/wpp/Download/Standard/Population/ 12 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 Global overweight and obese pregnant women 21. United Nations. World Population Prospects 2017. [cited 2017 Jun 23]. Available from: https://esa.un. org/unpd/wpp/Download/Standard/Fertility/. 22. World Health Organization. Global Health Observatory data repository. [cited 2017 Jun 23]. Available
from: http://apps.who.int/gho/data/node.main.A896?lang=en. PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 References Int J Envi-
ron Res Public Health. 2017; 14(8). https://doi.org/10.3390/ijerph14080898 PMID: 28792465. 37. Hosler AS, Michaels IH, & Buckenmeyer EM. Food Shopping Venues, Neighborhood Food Environ-
ment, and Body Mass Index Among Guyanese, Black, and White Adults in an Urban Community in the
US. Journal of Nutrition Education and Behavior. 2016; 48(6):361–8. https://doi.org/10.1016/j.jneb. 2016.03.003 PMID: 27085256. 38. Michimi A, Wimberly MC. The food environment and adult obesity in US metropolitan areas. Geospat
Health. 2015; 10(2):368. https://doi.org/10.4081/gh.2015.368 PMID: 26618317. 39. Trivedi T, Liu J, Probst J, Merchant A, Jhones S, Martin AB. Obesity and obesity-related behaviors
among rural and urban adults in the USA. Rural and remote health. 2015; 15(4):3267. PMID:
26458564. 40. Sheehy T, Sharma S. Trends in energy and nutrient supply in Trinidad and Tobago from 1961 to 2007
using FAO food balance sheets. Public Health Nutrition. 2013; 16(9):1693–1702. https://doi.org/10. 1017/S136898001200537X PMID: 23286774. 41. Sheehy T, Sharma S. The nutrition transition in the Republic of Ireland: trends in energy and nutrient
supply from 1961 to 2007 using Food and Agriculture Organization food balance sheets. British Journal
of Nutrition. 2011; 106(7):1078–89. https://doi.org/10.1017/S0007114511001395 PMID: 21481289. 42. Forsyth S, Gautier S, Salem Jr. N. Global Estimates of Dietary Intake of Docosahexaenoic Acid and
Arachidonic Acid in Developing and Developed Countries. Annals of Nutrition and Metabolism. 2016;
68:258–67. https://doi.org/10.1159/000446855 PMID: 27288396. 13 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 Global overweight and obese pregnant women 43. Vandevijvere S, Chow CC, Hall KD, Umali E, Swinburn BA. Increased food energy supply as a major
driver of the obesity epidemic: a global analysis. Bulletin of the World Health Organization. 2015; 93
(7):446–56. https://doi.org/10.2471/BLT.14.150565 PMID: 26170502. 44. Van Hook J, Altman CE, Balistreri KS. Global patterns in overweight among children and mothers in
less developed countries. Public Health Nutrition. 2013; 16(4):573–81. https://doi.org/10.1017/
S1368980012001164 PMID: 22583613. 45. Xu X, Variyam JN, Zhao Z, Chaloupka FJ. Relative Food Prices and Obesity in U.S. Metropolitan
Areas: 1976–2001. PLoS ONE. 2014; 9(12):e114707. https://doi.org/10.1371/journal.pone.0114707
PMID: 25502888. 46. Gonzalez-Gross M, Melendez A. Sedentarism, active lifestyle and sport: Impact on health and obesity
prevention. Nutr Hosp. 2013; 28 Suppl 5: 89–98. https://doi.org/10.3305/nh.2013.28.sup5.6923 PMID:
24010748. 47. Chu AH, Moy FM. Associations of occupational, transportation, household and leisure-time physical
activity patterns with metabolic risk factors among middle-aged adults in a middle-income country. Prev
Med. 2013; 57 Suppl:S14–7. https://doi.org/10.1016/j.ypmed.2012.12.011 PMID: 23276774. 48. 56.
Mi YJ, Zhang B, Wang HJ, Yan J, Han W, Zhao J, et al. Prevalence and Secular Trends in Obesity
Among Chinese Adults, 1991–2011. Am J Prev Med 2015; 49(5):661–9. https://doi.org/10.1016/j.
amepre.2015.05.005 PMID: 26275960. PLOS ONE | https://doi.org/10.1371/journal.pone.0202183
August 9, 2018 References Institute of Medicine (US) and National Research Council (US) Committee to Reexamine IOM Preg-
nancy Weight Guidelines. Weight Gain During Pregnancy: Reexamining the Guidelines. Washington
(DC): National Academies Press (US). 2009. 49. International Weight Management in Pregnancy (i-WIP) Collaborative Group. Effect of diet and physical
activity based interventions in pregnancy on gestational weight gain and pregnancy outcomes: meta-
analysis of individual participant data from randomised trials. BMJ. 2017; 358: j3119. https://doi.org/10. 1136/bmj.j3119 PMID: 28724518. 50. Yeo S, Walker JS, Caughey MC, Ferraro AM, Asafu-Adjei JK. What characteristics of nutrition and
physical activity interventions are key to effectively reducing weight gain in obese or overweight preg-
nant women? A systematic review and meta-analysis. Obesity Reviews. 2017; 18(4):385–99. https://
doi.org/10.1111/obr.12511 PMID: 28177566. 51. Lamminpaa R, Vehvilainen-Julkunen K, Schwab U. A systematic review of dietary interventions for ges-
tational weight gain and gestational diabetes in overweight and obese pregnant women. Eur J Nutr. 2017. https://doi.org/10.1007/s00394-017-1567-z PMID: 29128995. 52. Poston L, Bell R, Croker H, Flynn AC, Godfrey KM, Goff L, et al. Effect of a behavioural intervention in
obese pregnant women (the UPBEAT study): a multicentre, randomised controlled trial. Lancet Diabe-
tes Endocrinol. 2015; 3(10):767–77. https://doi.org/10.1016/S2213-8587(15)00227-2 PMID:
26165396. 53. Opie RS, Neff M, Tierney AC. A behavioural nutrition intervention for obese pregnant women: Effects
on diet quality, weight gain and the incidence of gestational diabetes. Aust N Z J Obstet Gynaecol. 2016; 56(4):364–73. https://doi.org/10.1111/ajo.12474 PMID: 27170563. 54. Seidell JC, Halberstadt J. The Global Burden of Obesity and the Challenges of Prevention. Annals of
Nutrition and Metabolism. 2015; 66 suppl 2:7–12. https://doi.org/10.1159/000375143 PMID:
26045323. 55. Pradeepa R, Anjana RM, Joshi SR, Bhansali A, Deepa M, Joshi PP, et al. Prevalence of generalized &
abdominal obesity in urban & rural India-the ICMR-INDIAB Study (Phase-I) [ICMR- NDIAB-3]. Indian J
Med Res. 2015; 142(2):139–50. https://doi.org/10.4103/0971-5916.164234 PMID: 26354211. 56. Mi YJ, Zhang B, Wang HJ, Yan J, Han W, Zhao J, et al. Prevalence and Secular Trends in Obesity
Among Chinese Adults, 1991–2011. Am J Prev Med 2015; 49(5):661–9. https://doi.org/10.1016/j. amepre.2015.05.005 PMID: 26275960. 14 / 14
|
https://openalex.org/W4319442556
|
https://www.frontiersin.org/articles/10.3389/fnut.2023.1106463/pdf
|
English
| null |
Prebiotic mechanisms of resistant starches from dietary beans and pulses on gut microbiome and metabolic health in a humanized murine model of aging
|
Frontiers in nutrition
| 2,023
|
cc-by
| 12,630
|
OPEN ACCESS EDITED BY
Peng-Gao Li,
Capital Medical University,
China
REVIEWED BY
Di Zhao,
Nanjing Agricultural University,
China
Jodi Woan-Fei Law,
Monash University Malaysia,
Malaysia
Tianming Yao,
Purdue University,
United States
*CORRESPONDENCE
Ravinder Nagpal
rnagpal@fsu.edu
SPECIALTY SECTION
This article was submitted to
Nutrition and Microbes,
a section of the journal
Frontiers in Nutrition
RECEIVED 23 November 2022
ACCEPTED 19 January 2023
PUBLISHED 07 February 2023
CITATION Saurabh Kadyan , Gwoncheol Park , Prashant Singh ,
Bahram Arjmandi and Ravinder Nagpal * Department of Nutrition and Integrative Physiology, College of Health and Human Sciences, Florida State
University, Tallahassee, FL, United States Department of Nutrition and Integrative Physiology, College of Health and Human Sciences, Florida State
University, Tallahassee, FL, United States Department of Nutrition and Integrative Physiology, College of Health and Human Sciences, Florida State
University, Tallahassee, FL, United States Dietary pulses, being a rich source of fiber and proteins, offer an ideal and inexpensive
food choice for older adults to promote gut and metabolic health. However, the
prebiotic effects of dietary pulses-derived resistant starches (RS), compared to RS
from cereals and tubers, remain relatively underexplored, particularly in context to
their gut modulatory potential in old age. We herein investigate the prebiotic effects
of pulses-derived RS on the gut microbiome and intestinal health in aged (60-week
old) mice colonized with human microbiota. C57B6/J mice were fed for 20 weeks
with either a western-style high-fat diet (control; CTL) or CTL diet supplemented (5%
w/w) with RS from pinto beans (PTB), black-eyed-peas (BEP), lentils (LEN), chickpeas
(CKP), or inulin (INU; reference control). We find that the RS supplementation
modulates gut microbiome in a sex-dependent manner. For instance, CKP enriched
α-diversity only in females, while β-diversity deviated for both sexes. Further, different
RS groups exhibited distinct microbiome differences at bacterial phyla and genera
levels. Notably, LEN fostered Firmicutes and depleted Proteobacteria abundance,
whereas Bacteroidota was promoted by CKP and INU. Genus Dubosiella increased
dominantly in males for all groups except PTB, whilst Faecalibaculum decreased
in females by CKP and INU groups. Linear discriminant analysis effect size (LEfSe)
and correlational analyzes reveal RS-mediated upregulation of key bacterial genera
associated with short-chain fatty acids (butyrate) production and suppression of
specific pathobionts. Subsequent machine-learning analysis validate decreased
abundance of notorious genera, namely, Enterococcus, Odoribacter, Desulfovibrio,
Alistipes and Erysipelatoclostridium among RS groups. CKP and LEN groups partly
protected males against post-prandial glycemia. TYPE Original Research
PUBLISHED 07 February 2023
DOI 10.3389/fnut.2023.1106463 TYPE Original Research
PUBLISHED 07 February 2023
DOI 10.3389/fnut.2023.1106463 TYPE Original Research
PUBLISHED 07 February 2023
DOI 10.3389/fnut.2023.1106463 COPYRIGHT
© 2023 Kadyan, Park, Singh, Arjmandi and
Nagpal. This is an open-access article
distributed under the terms of the Creative
Commons Attribution License (CC BY). The
use, distribution or reproduction in other
forums is permitted, provided the original
author(s) and the copyright owner(s) are
credited and that the original publication in this
journal is cited, in accordance with accepted
academic practice. No use, distribution or
reproduction is permitted which does not
comply with these terms. resistant starch, prebiotics, pulses, microbiota, gut dysbiosis 2.1. Extraction and preparation of resistant
starches from pulses Chickpeas, pinto beans, black-eyed peas and lentils were batch-
ordered and purchased from the commercial vendor1 for product
consistency and batch-tracking. Isolation and purification of starch
from pulse seeds were performed accordingly to our previously
described method (11). The recovered starch was dried in an oven
(45°C, 24 h) and stored under nitrogen at 4°C. The characteristics of
recovered pulse starch are shown in Supplementary Table S1. RS
production from purified starches was performed via simulation gastric
digestion as described previously by Tuncil et al. (18) with slight
modifications. Briefly, starch (12 g) was gelatinized in sodium
phosphate buffer (240 mL, pH – 6.9). After cooling to 37°C, 2 mL of
salivary amylase (Sigma-Aldrich) was added, followed by incubation
for 15 min. Under continuous stirring, the pH of the hydrolyzed starch
was adjusted from 6.9 to 2.0 using 6 M HCl. Thereafter, digestion was
carried out using sequential steps of enzymes: pepsin (37°C, pH 2.0,
30 min) and 4 mL pancreatin (37°C, pH 6.9, 90 min). The hydrolyzed
starch was recovered and dialyzed (6–8 kDa, 36 h) and the remaining
undigested starch was freeze dried for 72 h. Dietary pulses are an inexpensive, sustainable and valuable source
of nutrient-dense, health-promoting foods composed of both high-
quality proteins and dietary fibers, which could serve as a perfect-food
choice for the elderly in promoting gut and metabolic health (5). In fact,
a study, ‘Foods Habits in Later Life’ strongly predicted survival among
elderly consuming pulses (6). Among many pulse varieties grown
worldwide, chickpea, dry peas, lentils and dry beans, including black-
eyed peas, pinto beans and kidney beans, are some of the most widely
consumed pulses in the United States (7). Collectively, the consumption
of whole pulses has been shown to attenuate the risk of chronic diseases
by reducing the levels of post-prandial blood glucose, plasma levels of
low-density lipoprotein cholesterol, and blood pressure; however, the
underlying beneficial mechanisms of whole pulses and/or their
constituents on host physiology and gut microbiome in aging milieus
remain largely unexplored (8, 9). One such pulse-derived dietary fiber
worth exploring is the resistant starch (RS), which remains indigestible
in the upper gastrointestinal tract and is finally metabolized by intestinal
microbes in the large bowel, thereby promoting gut and metabolic
health (10). Apart from exerting health benefits, pulse-derived starches
could be a suitable ingredient for functional food formulations, as
studied earlier by our group (11). OPEN ACCESS Importantly, RS ameliorated high-
fat diet-induced gut hyperpermeability and enhanced expression of tight-junction
proteins (claudin-1 and claudin-4), which were more pronounced for LEN. In addition,
IL10 upregulation was more prominent for LEN, while TNF-α was downregulated by
LEN, CKP, and INU. Together, these findings demonstrate that RS supplementation
beneficially modulates the gut microbiome with a reduction in gut leakiness and
inflammation, indicating their prebiotic potential for functional food and nutritional
applications. Kadyan S, Park G, Singh P, Arjmandi B and
Nagpal R (2023) Prebiotic mechanisms of
resistant starches from dietary beans and
pulses on gut microbiome and metabolic
health in a humanized murine model of aging. Front. Nutr. 10:1106463. doi: 10.3389/fnut.2023.1106463 COPYRIGHT
© 2023 Kadyan, Park, Singh, Arjmandi and
Nagpal. This is an open-access article
distributed under the terms of the Creative
Commons Attribution License (CC BY). The
use, distribution or reproduction in other
forums is permitted, provided the original
author(s) and the copyright owner(s) are
credited and that the original publication in this
journal is cited, in accordance with accepted
academic practice. No use, distribution or
reproduction is permitted which does not
comply with these terms. resistant starch, prebiotics, pulses, microbiota, gut dysbiosis 01 frontiersin.org frontiersin.org Frontiers in Nutrition Kadyan et al. 10.3389/fnut.2023.1106463 Kadyan et al. Kadyan et al. 2.1. Extraction and preparation of resistant
starches from pulses The beneficial effects of RS extracted
from other sources, such as acorn and sago plants, in improving
physiological responses such as hyperglycemia and insulin sensitivity,
reducing gut hyperpermeability and systemic inflammation, and
modulating gut microbiome with the production of short-chain fatty
acids (SCFAs) had been demonstrated earlier (12). Likewise, the
physiologic and gut health effects of RS derived from cereal and tubers
have been widely studied, but studies pertaining to pulses-derived RS
have only started evolving recently (13–15). Furthermore, majority of
these studies utilize in-vitro fecal fermentation models, without much
extrapolation in in-vivo animal models. More importantly, the
mechanistic understanding of these pulses-derived RS among aging 1 https://nuts.com/cookingbaking/beans/ 1. Introduction population is lacking despite extensive use of dietary pulses as staple
foods worldwide. Aging is a predestined process of the human lifecycle, which is
usually accompanied by a gradual decline in physiological,
neurocognitive, and metabolic functions. The cumulative effect of
these reduced bodily functions weakens the immune system and
increases the susceptibility of the aged host to various
cardiometabolic, neurodegenerative, and lifestyle disorders. Emerging evidence is now pointing toward the crucial role of gut
microbiome homeostasis in host aging-associated health, implicating
the role of abnormal microbiome perturbations (‘dysbiosis’), induced
by factors such as increased medications and inadequate nutrition
with age, in exacerbating the pathophysiology of aging-linked
disorders (1, 2). Besides, increased consumption of western-style
high-sugar, high-fat and low-fiber diets during old age further
aggravates the ill effects of gut dysbiosis-linked aging-associated
ailments (3). Earlier studies by our group further corroborated the
link between the severity of gut dysbiosis with consumption of a
high-fat diet (HFD) in older non-human primates and the
amelioration of perturbed gut into a healthier microbiome upon the
intervention of high fiber diet, thus underscoring the incorporation
of fiber-rich diet in later life (4). As our gut microbiome coevolves
with our aging, strategic fiber-rich dietary interventions aimed at
maintaining and fostering a robust, diverse and healthier gut
microbial ecosystem in the elderly could indirectly prevent/offset the
progression of aging-associated diseases thereby potentially reducing
the economic burden associated with their treatment costs. Although human and mouse gut microbiotas share a 90%
resemblance at the phyla level, both possess dissimilarities in terms of
makeup and abundance of microbial signatures (16). Keeping in view
the emerging role of human intestinal microbiome in host physiologic
homeostasis, immunity, energy metabolism, and several disease
pathophysiologies, the current study was performed using a ‘humanized’
mouse model of aging, i.e., the older mice carrying the healthy human
gut microbiome, with the overarching aim to explore the prebiotic
attributes of RS purified from four of the most widely consumed pulses
(chickpea, black-eyed pea, pinto bean, and lentil) in modulating the gut
microbiome, physiological and intestinal health. The insights from this
novel study facilitate our understanding of developing nutraceuticals
from natural sources to support the health of our ever-growing elderly
population. On a broader perspective, this study will facilitate in
addressing the nutritional shortcoming of dietary fibers in the US as the
consumption of fibers in the country is less than 10% of recommended
intake (17). Frontiers in Nutrition 2.2. Animal studies The overall layout of the experimental design is summarized in
Figure 1A. Briefly, C57BL/6 J mice were purchased at ~55 weeks of age and
were allowed to acclimate in the new vivarium for 2 weeks. Then, mice were
subjected to our previously validated gut depletion and cleansing procedure
to eradicate the gut microbiota (12, 19). Briefly, the gut-cleansing procedure
included a 4-day microbial-depletion treatment with a broad-spectrum
antibiotic cocktail [containing ampicillin (1 g), Metronidazole (7–11) (1 g), 02 frontiersin.org 10.3389/fnut.2023.1106463 Kadyan et al. A
C
D
E
B
FIGURE 1
Effects of resistant starches from dietary pulses on bodyweight, diet intake, body-composition, total gastrointestinal length, colon length and cecum weight
in mice. (A) Experimental Design layout. (B) Overall trend depicting weekly change in bodyweight per group along with sex-specific changes in bodyweight
(%) and net weight gain (%). (C) Overall trend depicting weekly diet intake per group along with sex-specific average diet intake per week per group. (D) Overall and sex-specific alterations in body-composition (%) evaluated in terms of total fat and lean mass per group. (E) Comparison of total
gastrointestinal length, colon length and cecum weight of prebiotics with control. PTB, Pinto beans; BEP, black-eyed peas; LEN, Lentils; CKP, chickpeas;
INU, inulin; CTL, control high-fat diet group. #p < 0.1, *p < 0.05, **p < 0.01. B A B A C C D
E D C E E FIGURE 1
Effects of resistant starches from dietary pulses on bodyweight, diet intake, body-composition, total gastrointestinal length, colon length and cecum weight
in mice. (A) Experimental Design layout. (B) Overall trend depicting weekly change in bodyweight per group along with sex-specific changes in bodyweight
(%) and net weight gain (%). (C) Overall trend depicting weekly diet intake per group along with sex-specific average diet intake per week per group. (D) Overall and sex-specific alterations in body-composition (%) evaluated in terms of total fat and lean mass per group. (E) Comparison of total
gastrointestinal length, colon length and cecum weight of prebiotics with control. PTB, Pinto beans; BEP, black-eyed peas; LEN, Lentils; CKP, chickpeas;
INU, inulin; CTL, control high-fat diet group. #p < 0.1, *p < 0.05, **p < 0.01. CKP), and dry bean (Pinto; PTB), respectively. All diet formulations were
isocaloric (see Supplementary Table S2). 2.2. Animal studies After one-week, now at 60-weeks of age (matching to middle-aged
50–55 years old humans), the mice were randomly divided into five groups
(n = 14–16/group; 7–8 for each sex) and were fed a western-style high-fat
diet (HFD; #D21080102, Research Diets Inc., New Brunswick, NJ,
United States; formulated based on the USDA’s 2008 Dietary Assessment
of Major Food Trends) for 20-weeks. One group acted as control (CTL)
and received the western-style HFD, while four experimental groups
received CTL diet supplemented (5% w/w) with purified RS from dry peas
(Black-eyed pea; BEP), lentils (split lentil; LEN), chickpeas (Garbanzo; 2.2. Animal studies We have previously shown that
these four pulses, out of 18 different pulses, retain a considerable proportion
of RS pre- and post-cooking and lead to the production of short-chain fatty
acids (SCFAs) production by human gut microbiota in-vitro (11, 22). One
group received a widely studied fiber inulin (5% w/w; Biosynth) as a
positive control. The fortification proportion of 5% w/w and inclusion of
inulin as a positive control was based on our previous studies with
resistance starches and inulin (12). After the 20-week intervention, mice
were subjected to a series of assessments as described in the subsequent
sections. Mice were about 80–82 weeks old at this time, approximately
equivalent to age 60–65 years in humans. Body weight and diet intake were
measured weekly. At study endpoint, the mice were anesthetized with
isoflurane and euthanized. Cecum weight, liver weight and total
gastrointestinal length were measured. Tissues were collected in liquid
nitrogen and stored immediately at −80°C for further analysis. All the
animal studies and protocols were approved by the Florida State University’s
Institutional Animal Care and Use Committee (Protocol #202100008). Neomycin (1 g), and Vancomycin (0.5 g) per liter] in drinking water
(sweetened with 2% Medi-Drop Sucralose for palatability; Clear H2O,
Westbrook, ME), followed by bowel cleansing through 4-doses of oral
gavage with polyethylene glycol (200 μL per dose; 425 g/L) at 20-min
interval on the last day of antibiotic regimen to empty the intestine, wash
out the antibiotics remaining in the intestine, and decrease the bacterial
load. This regimen depletes more than 95% bacteria from the gut (12, 20,
21). Mice were fasted for 4 h before polyethylene cleansing. Because mice
are coprophagous, the cages and bedding materials were renewed each day
during this regimen. Then, mice were transplanted with a pool of five older
(3 M, 2F; age 50–55 year) subjects’ microbiota (collected recently in our lab;
IRB#00002132). To perform microbiota transfer, fecal contents from the
five human donors were pooled and dissolved in sterile PBS (100 mg/mL;
pH 7.4; pre-reduced in an anaerobic incubator), reconstituted in an
anaerobic chamber, and gavaged (200 μL/mice/day) for 4 consecutive days
(first FMT given right after 1 h after last polyethylene glycol treatment). For
the next one week, the mice were allowed to rest for microbiota stabilization. Frontiers in Nutrition frontiersin.org 2.6. In-vivo gut permeability assay Gut permeability was measured per our previously described
method (4, 12, 23). Briefly, mice were deprived of food for 4 hours, after
which the mice were administered with 60 mg/100 g body weight of
fluorescein isothiocyanate (FITC) dextran (4 kDa) solution via oral
gavage. Two hours after FITC gavage (still on fasting), about 50 μL of
blood was collected in a heparinized capillary tube by using a tail vein
cut. The concentration of the FITC-dextran was determined in the
serum using fluorescence spectroscopy at 530 nm and excitation at
485 nm using a plate reader. 3. Results 2 https://earthmicrobiome.org/ 2.8. Statistical analysis All the values throughout the manuscript are expressed as
mean ± standard error of the mean (SEM). Statistical significance
between groups was assessed using a two-tailed unpaired t-test using
Welch’s correction in the GraphPad Prism version 9.4.1. Microbiome
analyzes were executed using ‘R’ or ‘Python’ packages. The
microbiome diversity indices (α- and β-diversity), as well as
microbiome composition in terms of major phyla, families and
genera, were compared between control and different treatment
groups. Non-parametric Kruskal-Wallis test (34) and PERMANOVA
(35) with 999 random permutations were used to identify the
significant differences in microbial diversity and structure. All
p-values involving multiple testings were FDR-adjusted. Supervised
classification was performed in the q2-sample-classifier plugin via
nested stratified 5-fold cross-validation with Random Forest (36)
classifier grown with 2,000 trees. The metagenomic functional
activities were predcted using the open source bioinformatics tool
PICRUSt2 (Phylogenetic Investigation of Communities by
Reconstruction of Unobserved States) (37) and output were annotated
with KEGG Brite descriptions (38). The sequences were uploaded to
PICRUSt2 and were analyzed for the prediction of functional genes
of the classified members of the gut microbiota. Subsequently, the
inferred gene families were annotated against KEGG (Kyoto
encyclopedia of genes and genomes) orthologs (Kos) and then
collapsed into KEGG pathways to generate the functional pathway. The functions were finally categorized and compared at levels 2 and
3 as per the methods described elsewhere (37). The linear discriminant
analysis (LDA) effect size (LEfSe) (39) was used to identify the
difference in bacterial taxa and predicted functions between groups. Additionally, STAMP v 2.1.3 software (40) and Spearman’s correlation
were used to detect the group-specific taxa. InteractiVenn (41) was
used to illustrate the Venn diagram. Unless otherwise specified,
statistical significance was tested at p < 0.05. 2.3. Body composition Total body composition in terms of lean mass versus fat mass (and
total water) at the study endpoint was determined in live mice by using
the EchoMRI-130 Body Composition Analyzer (EchoMRI, MRI that
Counts, Houston, TX). Frontiers in Nutrition 03 Kadyan et al. 10.3389/fnut.2023.1106463 10.3389/fnut.2023.1106463 Kadyan et al. Kadyan et al. 2.4. Gut microbiome analysis transcription using the High-capacity cDNA reverse transcription kit
(ThermoFisher). The mRNA expression of tight-junction proteins,
including claudin-1 (CLDN1), claudin-4 (CLDN4), zonulin-1 (ZO1),
zonulin-2 (ZO2), occludin (OCCL) and JAM3 gene; and inflammatory
markers including interleukins (IL1β, IL10, and IL17A) and tumor
necrosis factor-alpha (TNF-α) were quantified using real-time PCR
(QuantStudio3, Applied Biosystems) with primers listed in
Supplementary Table S3. The 18S gene was used as an internal
housekeeping control. The results were expressed as ddCt method by
normalizing against the 18S expression of the control group, as per our
earlier report (12, 23). The gut microbiome will be measured as per our previously described
methods (4, 12, 23–27). Briefly, genomic DNA from 200 mg of the fecal
specimen was extracted using the QIAmp PowerFecal Pro DNA Kit
(Qiagen) according to the manufacturer’s instructions. Universal primers
515F (barcoded) and 806R were used to amplify the hypervariable V4
region of the bacterial 16S rRNA gene in accordance with the Earth
Microbiome Project benchmark protocol2. The resulting amplicons were
purified via AMPure® magnetic purification beads (Agencourt) followed
by quantification through Qubit-4 fluorimeter (InVitrogen), and the final
amplicon library was generated as per methods disclosed elsewhere. Equal molar concentrations of the library were pooled and sequenced for
paired-end (2 × 300bp) sequencing using an Illumina MiSeq sequencer
(using Miseq reagent kit v3; Illumina Inc., San Diego, United States). QIIME2 (ver. 2–2022.8) was used for microbiome bioinformatics analysis
(28). Raw sequence demultiplexing and filtering on quality was
performed with the q2-demux plugin, followed by trimming and
denoising through DADA2 (29). All identified amplicon sequence
variants (ASVs) were aligned with the MAFFT (30). ASVs were assigned
with a naïve Bayes taxonomy classifier developed for the sklearn classifier
against the pre-built from the 99% SILVA 138 database (31, 32). 2.5. Meal tolerance test and insulin tolerance
test These assessments were measured as per our previously described
methods (4, 12, 33). Briefly, the meal tolerance test (MTT) was
performed by oral gavage of 200 μL mixed meal (570 mg Ensure Plus,
30 mg dextrose, and 1.8 mL water) to mice fasted for 6 h. For the insulin
tolerance test (ITT), mice fasted for 4 h and were given 0.75 U/kg body
weight of insulin (Humulin) intra-peritoneally. Blood glucose reading
for both tests was measured at 0 min (before administration), and 15, 30,
60, and 120 min (after administration) using AccuCheck glucometer kit. Frontiers in Nutrition 3.2. Resistant starches from different beans
and pulses differently modulate the gut
microbiome The overall and sex-specific impact of RS on the α-diversity of the
gut microbial community was depicted using Shannon and Chao1
index, as depicted in Figures 2A,B. No significant differences in
α-diversity were observed overall, but CKP significantly enhanced
α-diversity in females compared to CTL as revealed by Shannon index
(PFDR = 0.022). However, the β-diversity revealed notable differences in
the microbial structure between CTL and RS-treated groups. Both INU
and CKP caused significant structural changes in both sexes, whereas
BEP (PFDR = 0.011) and LEN (PFDR = 0.004) induced differences only in
males (Figure 2C). Besides, sex-specific effects of different RS on
β-diversity were observed, as presented in Figure 2D. Compared to the
CTL group, the overall proportion of major bacterial phyla including
Firmicutes and Bacteroidota did not differ between groups, but the LEN
group harbored a significantly higher proportion of Firmicutes (male-
driven) while Bacteroidota (female-driven) was more abundant in the
CKP and INU groups (Figure 2E). Furthermore, phyla Proteobacteria
and Actinobacteria decreased by LEN and INU intake, respectively. Further analysis at the bacterial genus level revealed Faecalibaculum
(female-dominated), Dubosiella (male-dominated), Enterococcus, and
Bacteriodes as dominant genera accounting for 50–60% of total
abundance across different groups (Figure 2F). Overall, LEN had a
maximum abundance of former two genera. Interestingly,
Faecalibaculum decreased in CKP and INU. Enterococcus was found in
more abundance in PTB but was reduced in other groups compared to
CTL. INU had the highest rise for Bacteriodes. A detailed description of
hierarchical clustering at bacterial phylum, family, and genus levels for
individual samples is presented in Supplementary Figure S2. Colidextribacter, Holdemania and Lachnospiraceae;g_ were
significantly associated with CKP, while Parasutterella and Blautia were
linked with INU group. Correlation analyzes between control and
treatment groups for the top 20 genera are summarized in Figure 4B. The
genera exhibiting significantly positive and negative correlations among
different groups versus control were nearly identical to those found
significantly different by LEfSe analyzes, except for CKP, which showed
additional insights for genera associated negatively (Eggerthella and
Enterococcus) and positively (Desulfovibrio, Lactococcus, Parvibacter,
besides others) correlated. In general, positively (Lachnospiraceae;g_
uncultured, Ruminococcaceae;g_uncultured, Flavonifractor and A2) and
negatively correlated genera (Bifidobacterium, Eggerthella, CAG-532,
Enterobacteriaceae;g) shared common features in at least three of RS
groups compared to CTL. KEGG-based functional analysis annotated
these microbiome changes to a total of 12 differential metabolic
pathways when compared to CTL (Figure 4). 3.1. Effects of dietary pulses-derived
resistant starches on physiological
parameters Total RNA from the snap-frozen intestinal tissues (colon and ileum)
were isolated using RNeasy kit (Qiagen), followed by reverse The overall effect of RS on body-weight, diet intake, body-
composition, intestinal/colon length, and cecum weight are summarized
in Figures 1B–E. There was no marked impact of different RS on the
body-weight of either male or female mice. Although non-significant, 04 frontiersin.org frontiersin.org 10.3389/fnut.2023.1106463 10.3389/fnut.2023.1106463 Kadyan et al. demonstrated a significantly higher proportion of Lachnospiraceae,
Clostridium sensu stricto-1, f_Ruminococcaceae and Faecalitalea while
having a lower proportion of Desulfovibrio, Lactococcus, Bifidobacterium
and Dubosiella. In BEP group, Lachnospiraceae, Ruminococcaceae,
Oscillospiraceae, and Flavonifractor increased, while Eubacterium
fissicatena group, Eggerthella, CAG_352, Bifidobacterium and
Enterococcus decreased. The LEN group showed relatively least
significant
difference
compared
to
the
CTL
group
with
Christensenellaceae R-7 group deceased and Lachnospiraceae UCG-004
increased compared to CTL. CKP exhibited an increase in 12 genus-
specific taxa compared to CTL, which included Bacteroides,
Lachnospiraceae, Oscillospiraceae, Ruminococcaceae-g_Incertae_Sedis,
Colidextribacter, besides several other taxa, whereas assigned taxa
including Christensenellaceae R-7 group and Faecalibaculum decreased
significantly. For INU group, Dubosiella, Bacteriodes, Blautia and
Lachnospiraceae were among the selected taxa that increased and
Ruminococcaceae, Streptococcus, Christensenellaceae_R_7_g, and
Faecalibaculum were among the decreased ones. the INU group demonstrated less weight gain overall, whereas PTB and
BEP groups showed restricted weight gain in females and males,
respectively (Figure 1B). There was no difference in the food intake in
either gender (Figure 1C). Further, no notable differences were observed
between fat and lean mass of control and experimental groups; however,
male mice from PTB group exhibited marginally lower fat mass gain
(p = 0.0781) compared to CTL (Figure 1C). Overall, the intestinal length
was significantly longer for PTB and INU groups whereas an extension
in colon length was observed for BEP group (Figure 1E) mainly in males
(Supplementary Figure S1). Moreover, BEP (p < 0.05) and INU
(p < 0.0001) had significantly heavier cecum than CTL. INU group-
associated increase in cecum weight was consistent in both sexes
(Supplementary Figure S1). No overall difference in liver weight was
observed, but PTB group females demonstrated low liver weight
(p < 0.05) compared to CTL (Supplementary Figure S1). Altogether, no
single group yielded consistent results for these parameters, although
some overlapping effects were observed for INU group in both sexes,
PTB in females, and BEP in males. 3.1. Effects of dietary pulses-derived
resistant starches on physiological
parameters To identify discriminatory taxa associated with each RS, we applied
a supervised machine learning-based random forest model trained with
genus-level abundance. The model demonstrated a high classification
accuracy for each RS (AUC = 0.70–0.95) and the overall accuracy of
prediction was 60.4% (Supplementary Figures S3A,B). Among the 25
most predictive genera influenced by RS, 16 genera showed a significant
difference between CTL versus at least one of four RS groups
(Figure 4A). The prominent predictive genera included Faecalibaculum,
Lactococcus, Enterococcus, Streptococcus, Dubosiella, Blautia and
Bacteroides. The presence of these top genera, along with Odoribacter,
Desulfovibrio, Lactobacillus, Alistipes and Erysipelatoclostridium was
predominant in CTL group. Except for PTB, Dubosiella was more
prevalent in other groups compared to CTL. Bacteroides predicted more
strongly towards CKP and INU groups. Frontiers in Nutrition 3.2. Resistant starches from different beans
and pulses differently modulate the gut
microbiome PTB, Pinto beans; BEP, black-eyed peas; LEN, Lentils; CKP, chickpeas; INU, inulin; CTL, control high-fat
diet group. *p < 0.05. 3.3. Resistant starches from different dietary
pulses restrict elevation in post-prandial
blood glucose levels independent of insulin
sensitivity 3.2. Resistant starches from different beans
and pulses differently modulate the gut
microbiome Overall, pathways
pertaining to amino acid, vitamins, carbohydrate, glycan, and lipid
metabolism were upregulated prolifically in all RS groups
(Supplementary Figure S4). Pathways specific to KEGG level-3 that
upregulated in specific treatment groups included histidine metabolism
(BEP); and sphingolipid metabolism and glycan synthesis (INU). On the
other side, downregulated pathways were linked with glycolysis
gluconeogenesis (BEP, CKP, INU); xenobiotics degradation (LEN);
galactose metabolism (CKP); biosynthesis of peptidoglycan and
terpenoids (INU); metabolism of D-alanine, D-glutamine, and
nucleotides (INU). Among total 1,898 amplicon sequence variants (ASVs) obtained, the
largest number of unique ASVs (280, 14.8%) were identified in the CKP
group, followed by BEP (235, 14.16%) and LEN (219, 10.59%)
(Figures 3A,B). The identification of characteristic bacterial taxa changes
at genus and various taxonomic levels were further visualized using the
linear discriminant analysis (LDA) scores and LDA effect size (LEfSe)-
based cladogram analyzes, respectively (Figures 3C,D). PTB group 05 frontiersin.org 10.3389/fnut.2023.1106463 Kadyan et al. A
C
B
D
E
F
FIGURE 2
Effect of resistant starches from dietary pulses on gut microbiome diversity and taxonomic hierarchies. Alpha-diversity of microbiome of each group
determined using (A) Shannon index and (B) Chao1 index. (C) PCoA analysis representing β-diversity of microbiome of each group combined or male and
female groups. (D) Sex-specific differences in microbiome composition for each group; covariance ellipses are displayed and bounds (dotted lines) of each
group with two standard deviations (2σ) in each direction from the group mean. Relative abundance of gut microbiome in male and female mice or all
combined at (E) phylum level and (F) genus level. PTB, Pinto beans; BEP, black-eyed peas; LEN, Lentils; CKP, chickpeas; INU, inulin; CTL, control high-fat
diet group. *p < 0.05. C
D A
B C
D C A D B B D
F B F E E F E
F FIGURE 2
Effect of resistant starches from dietary pulses on gut microbiome diversity and taxonomic hierarchies. Alpha-diversity of microbiome of each group
determined using (A) Shannon index and (B) Chao1 index. (C) PCoA analysis representing β-diversity of microbiome of each group combined or male and
female groups. (D) Sex-specific differences in microbiome composition for each group; covariance ellipses are displayed and bounds (dotted lines) of each
group with two standard deviations (2σ) in each direction from the group mean. Relative abundance of gut microbiome in male and female mice or all
combined at (E) phylum level and (F) genus level. 3.3. Resistant starches from different dietary
pulses restrict elevation in post-prandial
blood glucose levels independent of insulin
sensitivity oral meal and intra-peritoneal insulin administration, are depicted in
Figures 5A,B. Overall, BEP and CKP groups yielded lower blood glucose
after 30 and 60 min of meal administration, respectively (Figure 5A). Besides, males from the CKP group presented the lowest AUC
(p = 0.0782) for blood glucose. Although no significant variations in
meal tolerance were observed for females, LEN group in males rendered
a relatively lower yet stable trend towards change in post-prandial The overall changes in blood glucose levels, along with sex-specific
percent changes in glucose and area under the curve (AUC) during the 06 Frontiers in Nutrition frontiersin.org 10.3389/fnut.2023.1106463 Kadyan et al. A
B
C
D
FIGURE 3
Comparison of unique microbiome features and discriminatory taxa between prebiotics (resistant starches and inulin) versus control groups. (A) Venn
diagram illustrating unique and shared features (ASVs) among different groups. (B) Unique features expanded to display top 10 genera with relative
abundance of unique features of each group. (C) Bacterial composition differences between groups analyzed with Linear discrimination analysis (LDA)
effect size (LefSe) algorithm; x-axis: LDA score. (D) LEfSe cladogram depicting differentially abundant taxa between groups. PTB, Pinto beans; BEP,
black-eyed peas; LEN, Lentils; CKP, chickpeas; INU, inulin; CTL, control high-fat diet group. A
B B A A C C D FIGURE 3
Comparison of unique microbiome features and discriminatory taxa between prebiotics (resistant starches and inulin) versus control groups. (A) Venn
diagram illustrating unique and shared features (ASVs) among different groups. (B) Unique features expanded to display top 10 genera with relative
abundance of unique features of each group. (C) Bacterial composition differences between groups analyzed with Linear discrimination analysis (LDA)
effect size (LefSe) algorithm; x-axis: LDA score. (D) LEfSe cladogram depicting differentially abundant taxa between groups. PTB, Pinto beans; BEP,
black-eyed peas; LEN, Lentils; CKP, chickpeas; INU, inulin; CTL, control high-fat diet group. Frontiers in Nutrition 3.4. Resistant starches from different dietary
pulses ameliorate gut permeability and leaky
gut markers glucose levels compared to CTL. Regarding insulin sensitivity, no
significant differences were observed among control and treatment
groups in male or female mice (Figure 5B), although a marginal decrease
in AUC for CKP group males might suggest improved insulin sensitivity. Overall, CKP and LEN groups offered protection to some extent against
post-prandial glucose levels in males. The influence of RS on intestinal epithelial layer integrity in both
males and females was examined in terms of the mRNA expression of 07 frontiersin.org 10.3389/fnut.2023.1106463 Kadyan et al. A
B
C
FIGURE 4
Comparison of predictive microbial genera, functional profiles and correlation analysis between resistant starches and control groups. (A) The top 25 most
strongly predictive genera ordered by relative importance score used to assess the contribution to classifier accuracy, and extended error bar plots with
corrected value of p (Welch’s two-sided t-test) for each taxon. (B) Correlation analysis for top 20 genera. (C) Differences in predictive KEGG functional
capabilities using the LEfSe (LDA > 3.0) analysis. PTB, Pinto beans; BEP, black-eyed peas; LEN, Lentils; CKP, chickpeas; INU, inulin; CTL, control high-fat diet
group. *p < 0.05. A B B
C C FIGURE 4
Comparison of predictive microbial genera, functional profiles and correlation analysis between resistant starches and control groups. (A) The top 25 most
strongly predictive genera ordered by relative importance score used to assess the contribution to classifier accuracy, and extended error bar plots with
corrected value of p (Welch’s two-sided t-test) for each taxon. (B) Correlation analysis for top 20 genera. (C) Differences in predictive KEGG functional
capabilities using the LEfSe (LDA > 3.0) analysis. PTB, Pinto beans; BEP, black-eyed peas; LEN, Lentils; CKP, chickpeas; INU, inulin; CTL, control high-fat diet
group. *p < 0.05. significant regulation in males’ colon but found to increase in the ileum
by PTB and INU groups (Figure 6D). Furthermore, we also measured
gut leakiness by gut permeability assay using oral FITC-dextran
administration in fasted mice, the findings of which are depicted in
Figures 6E,F. Overall, PTB and INU reduced gut leakiness in both sexes
compared to CTL. BEP and LEN had significantly improved gut
permeability only in females (Figure 6E). Unexpectedly, CKP group
males had increased gut permeability, with no major reduction
observed in females. Frontiers in Nutrition frontiersin.org 3.4. Resistant starches from different dietary
pulses ameliorate gut permeability and leaky
gut markers This finding provides a divergent result compared
to leaky gut expression analysis as the positive effect CKP on increased
expression of barrier-forming genes (CLDN1 and CLDN4) in males
was observed. different barrier-forming and scaffolding genes in ileum and colon
tissues (Figures 6A–D). Among females, LEN group increased CLDN-1
expression in both tissues while expression of OCCL and JAM3
decreased in the ileum with no major impact on the colon
(Figures 6A,C). The scaffolding proteins (ZO1 and ZO2) mostly
remained unaffected in the colon of females, whereas all groups except
CKP downregulated ZO1 expression in the ileum. Moreover, PTB
group enhanced CLDN-1 expression in the ileal tissues of both male
and female mice (Figures 6C,D). INU group exerted male-specific
enhanced expression of CLDN-1 and OCCL in the colon and ileum
(Figures 6B,D). CLDN-4 was upregulated by PTB, LEN and CKP
groups in the males’ colon (Figure 6B). While ZO2 exhibited no 08 Frontiers in Nutrition frontiersin.org Kadyan et al. 10.3389/fnut.2023.1106463 A
B
FIGURE 5
Effect of resistant starches from dietary pulses on post-prandial blood glucose levels and insulin sensitivity. (A) Meal tolerance test and (B) Insulin tolerance
test in combined or separately in males and females along with sex-specific percent change in blood glucose levels and area under curve (AUC). PTB, Pinto
beans; BEP, black-eyed peas; LEN, Lentils; CKP, chickpeas; INU, inulin; CTL, control high-fat diet group. #p < 0.1, *p < 0.05. A A B B B FIGURE 5
Effect of resistant starches from dietary pulses on post-prandial blood glucose levels and insulin sensitivity. (A) Meal tolerance test and (B) Insulin tolerance
test in combined or separately in males and females along with sex-specific percent change in blood glucose levels and area under curve (AUC). PTB, Pinto
beans; BEP, black-eyed peas; LEN, Lentils; CKP, chickpeas; INU, inulin; CTL, control high-fat diet group. #p < 0.1, *p < 0.05. frontiersin.org Frontiers in Nutrition 3.5. Resistant starches from different dietary
pulses differentially influence inflammatory
markers in males and females Dietary modulation of the gut microbiome to ameliorate
pathophysiology of various aging-associated non-communicable
diseases such as obesity and type-2 diabetes is receiving remarkable
attention. Among these modulatory approaches, dietary fibers,
including resistant starches, could be a promising ingredient for gut
health owing to their ability to safely pass through the upper
gastrointestinal tract and act as good substrates for fermentation by
colonic microflora. To this end, the current investigation evaluated the
prebiotic potential of resistant starches extracted from the different
dietary pulses, viz. pinto beans, black-eyed peas, lentils, and chickpeas
on the gut and metabolic health outcomes in a ‘humanized’ mouse
model of aging. RS supplementation within a high-fat diet (HFD)
induced distinct sex- and starch-specific alterations in the gut
microbiome, post-prandial hyperglycemia, leaky gut, and inflammatory
markers in human microbiome-harboring mice. The overall effect of
RS supplementation on weight gain did not yield significant differences
compared to CTL till 12-weeks. Although the protective effect of RS
type-2 (RS2) on high-fat diet has previously been demonstrated earlier
(42), weight reduction in some studies was not evident (43, 44). Moreover, a recent study reported detectable weight reduction after The overall gender-specific impact of RS in modulating the
expression of pro-inflammatory (IL1β, IL17A, and TNFα) and anti-
inflammatory (IL10) cytokines are presented in Figures 6G–J. In
females, IL10 and IL17A increased consistently in the colon and ileum
upon LEN treatment, while PTB supplementation increased IL10 in the
colon (Figures 6G,I). In males, IL10 was increased by LEN in the colon
and INU in the ileum (Figures 6H,J). In females, LEN significantly
downregulated TNFα expression in the ileum along with its
non-significant reduction in the colon by CKP (Figures 6G,I). For
males, a decrease in TNFα was observed in the colon by CKP and INU
groups, while a BEP-mediated increase occurred in the ileum
(Figures 6H,J). The decline in IL1β expression by INU group was
evident in the ileum of both males and females and in the colon of
males (Figures 6H–J). Altogether, RS supplementation enhanced anti-
inflammatory cytokine (IL10) and declined pro-inflammatory (TNFα)
response in both sexes. In addition, a positive correlation was observed
between IL10 and IL17A cytokines levels, which was strikingly evident
for the LEN group. 3.5. Resistant starches from different dietary
pulses differentially influence inflammatory
markers in males and females 09 Frontiers in Nutrition Kadyan et al. 10.3389/fnut.2023.1106463 A
C
E
B
G
H
I
J
D
F
FIGURE 6
Effect of resistant starches from dietary pulses on the markers of gut permeability, intestinal integrity, and inflammation. Gene expression of tight-junction
proteins viz., claudins (CLDN-1 and CLDN-4), zonulins (ZO1- and ZO-2), occludin (OCCL), and JAM3 in (A) females’ colon, (B) males’ colon, (C) females’
ileum and (D) males’ ileum. Gut permeability (in terms of the diffusion of 4 kDa FITC dextran from gut to blood circulation) in (E) females and (F) males. Gene expression of inflammatory markers including interleukins (IL1β, IL10, and IL17) and tumor necrosis factor-alpha (TNFα) in (G) females’ colon,
(H) males’ colon, (I) females’ ileum and (J) males’ ileum. PTB, Pinto beans; BEP, black-eyed peas; LEN, Lentils; CKP, chickpeas; INU, inulin; CTL, control
high-fat diet group. *p < 0.05, **p < 0.01. E
F A
C
B
D C
E
D
F E C A F D
F B D B
D F G
H
FIGURE 6 H J I G I
J J Effect of resistant starches from dietary pulses on the markers of gut permeability, intestinal integrity, and inflammation. Gene expression of tight-junction
proteins viz., claudins (CLDN-1 and CLDN-4), zonulins (ZO1- and ZO-2), occludin (OCCL), and JAM3 in (A) females’ colon, (B) males’ colon, (C) females’
ileum and (D) males’ ileum. Gut permeability (in terms of the diffusion of 4 kDa FITC dextran from gut to blood circulation) in (E) females and (F) males. Gene expression of inflammatory markers including interleukins (IL1β, IL10, and IL17) and tumor necrosis factor-alpha (TNFα) in (G) females’ colon,
(H) males’ colon, (I) females’ ileum and (J) males’ ileum. PTB, Pinto beans; BEP, black-eyed peas; LEN, Lentils; CKP, chickpeas; INU, inulin; CTL, control
high-fat diet group. *p < 0.05, **p < 0.01. 12-weeks of intervention, suggesting delayed therapeutic effects of RS2
(3). Also, the anti-obesity effects of RS are dose-dependent with diets
containing at least 8% RS exhibiting reduced adiposity irrespective of
phenotype (45). We also observed statistically insignificant but
noticebale sex-specific restricted weight gain for PTB and BEP groups. Sex-specific differences can also affect the trajectory of weight gain as
studied previously by feeding RS (a retrograded amylose starch)
supplemented diet to human microbiome-harboring rats (46). The
terminal bodyweight of male rats was increased while that of females
either remain unaffected or decreased after RS consumption. Frontiers in Nutrition frontiersin.org 3.5. Resistant starches from different dietary
pulses differentially influence inflammatory
markers in males and females RS-mediated increases in cecum weight have been reported earlier (43),
which may reflect the enhanced fermentative ability of gut microbes
guided by dietary fibers (47). RS-enriched diets have been shown to
promote gastrointestinal length, which, especially for colon, may also
reflect on the proportion of RS entering the colon (48). Besides, an
extension in colon length is also considered as a useful indicator of reduced intestinal inflammation associated with improved barrier
function, mucosal repair, and enterocyte hyperplasia (49). Compelling evidence has demonstrated the modulating role of RS
in the restructuring of gut microbiome composition, which is further
dictated by the selective ability of gut microbes to metabolize different
RS types (50). In the present study, α- and β-diversity arrays were
affected differently by the consumption of RS from different sources,
with CKP demonstrating a more pronounced effect overall. Such
differences could be ascribed to structural variations in RS linked with
different legume cultivars (5). Whole chickpea powder supplemented in
an obesogenic diet has previously been reported to yield superior
α-diversity compared to other pulse groups (9). Likewise, we noted that
different RS groups exhibit differential effects across all taxonomic
hierarchies, which further vary by gender. LEN exhibited positive effects
at the phylum level by enhancing Firmicutes (predominant in males)
and depleting Proteobacteria. These results provide divergent findings 10 10.3389/fnut.2023.1106463 10.3389/fnut.2023.1106463 Kadyan et al. as Firmicutes are generally decreased after consumption of whole lentils
in a high-fat diet (9), signifying the interactive effects of other lentil
components in the diet. Increased abundance of Firmicutes at the
expense of Bacteroidetes in individuals receiving RS2 has also been
reported previously (51). Many Firmicutes members are associated with
butyrate production, whereas Proteobacteria encompass many
lipopolysaccharides (LPS)-producing species which may induce gut
dysbiosis and the onset of several diseases (3, 52). Besides, Bacteroidota
(predominant in females), which increased after CKP and INU
treatment, harbors genera having the ability to degrade complex
carbohydrates, including RS, to produce acetate and propionate (52). The influence of gender in orchestrating gut microbiome composition
has received considerable attention lately as microbiota differences
among sexes could contribute to variation in local and systemic
inflammation as well as susceptibility to an array of cardiometabolic
diseases. Studies have emphasized to consider gender-specific effects
while understanding the interactions between gut microbiome and diet
(53). 3.5. Resistant starches from different dietary
pulses differentially influence inflammatory
markers in males and females However, the
growth of Bifidobacterium and Lactobacillus may also be affected by the
fat content in RS-supplemented diet (71). Nonetheless, the response of
gut microbiome to RS fermentation is quite complex, which may
be affected by nuanced structural and physicochemical variations in RS
derived either from same or different botanical origin (72). Studies have
attributed microstructural features of RS such as amylose content,
degree of crystallinity (B-type), chain-length and branching density,
blocklet type, surface porosity and texture as a principal factors behind
enhanced resistance of RS to digestion and its subsequent differential
modulation of gut microbiome (73, 74). Depending upon the source,
retrogradation process and amylose content of native starches, formation
of RS3 with B-type polymorphic form, low surface porosity, having a
compact and order blocklet structure along with higher proportion of
chains with moderate degree of polymerization (37–100) could act as
superior prebiotic for gut and metabolic health, as investigated recently
by (75). Even though mining these physicochemical structural variations
in RS was not the prime focus of this study, accumulating evidence
points out the governing role of fine structural variations of RS in
modulating the gut microbiome differently, which may be investigated
in future to delve deep into targeted microbiota interventions. The ASVs of each treatment group retrieved from 16S rRNA
i
i ld d lt
ti
i th i
t b li
th
d The specific effects of RS in increasing the abundance of some
reported SCFAs producers, viz., Faecalibaculum, Dubosiella, and
Bacteriodes and reducing obesity-associated genera (e.g., Enterococcus)
corroborate well with recent studies (14, 56). Moreover, Enterococcus,
which had high relative abundance in CTL group of males, was not
suppressed by PTB intervention. However, this consequence exhibited
no detectable adverse outcomes in terms of glucose response,
inflammation, and gut leakiness for PTB group in males. Even though
some species of Enterococcus are opportunistic pathogens, many others
are known to be beneficial or commensal to the host. For instance,
E. hirae has been reported to enhance anti-cancer immune responses
and improve gut epithelial integrity (57). Other studies have also
reported increased Enterococcus levels in rats by consumption of high-
amylose maize starch and its protective role in reducing bile acid
toxemia by prompting the excretion of primary bile acids from the
intestine (58). 3.5. Resistant starches from different dietary
pulses differentially influence inflammatory
markers in males and females Previous study by our group established gender-specific
microbiome changes upon yoghurt consumption, wherein consumption
patterns in male cohorts were negatively correlated with
Lactilactobacillus sakei, Staphylococcus and Enterobacteriaceae, whereas
Lacticaseibacillus casei found positively correlated with females (54). Sex-specific alterations in the gut community structure of rats receiving
prebiotic (oligofructose)-enriched chow have also been documented
(55), wherein female rats exhibited predominance of phylum
Bacteroidetes while no major changes in fecal community structure of
males were observed. This highlights the importance of including both
sexes during dietary intervention studies involving animal models to
fully elucidate the universality of health benefits imparted by dietary
fibers and prebiotics. various potential pathobionts among several discriminatory taxa
identified using a machine-learning approach, such as Odoribacter,
Alistipes and Erysipelatoclostridium were found more abundant in CTL
group, further highlighting the potential of RS in suppressing potentially
harmful gut microbial inhabitants. Previous studies have also
demonstrated the regulatory role of RS in inhibiting the growth of
Erysipelatoclostridium, Desulfovibrio, Alistipes (aging-associated taxa)
and Odoribacter (adiposity-linked taxa) in HFD-fed mice (3, 63, 64). In
a recent study, the abundance of Streptococcus in CTL compared to RS
groups was reported, which could be attributed to obesity-induced
inflammatory responses (65). Christensenellaceae R-7 group, which
exhibited a negative correlation with LEN and INU groups, has also
been reported to be inhibited by high amylose content in RS-enriched
diets (66). Members of the Christensenellaceae family are associated with
reduced adiposity in general and are specialized in protein fermentation
(67). Recent studies on [Eubacterium]_fissicatena_g (negatively
correlated with BEP) showed its close association with hypertension
(68). However, the same has also been linked to reduced body weight
and adiposity (69). Nevertheless, the current knowledge on the clinical
importance of these two genera with respect to metabolic disorders is
still limited and needs further investigation. Considering the ambiguous
role of some genera (e.g., Enterococcus, Eubacterium) on host
metabolism, further species level identification using shotgun
metagenomics would provide more information on the potential
mechanisms of RS on gut health as a function of species within a
particular genus. Although Bifidobacterium was negatively correlated
with RS groups in the current study, it was not among the top 25
discriminatory taxa associated with each RS by the machine learning
algorithm. This is in contrary to the previous studies reporting an
increased prevalence of Bifidobacterium in rodent and in-vitro
fermentation models following RS intervention (13, 70). Frontiers in Nutrition frontiersin.org 3.5. Resistant starches from different dietary
pulses differentially influence inflammatory
markers in males and females The Ruminococcaceae, Oscillospiraceae, and Lachnospiraceae families
among Firmicutes phylum harbor varied SCFAs-producing genera, with
many of their uncultured members observed to be enriched in RS
groups as revealed by the LEfSe analysis (59). Lachnospiraceae_
UCG_004, a genus abundant in the LEN group, has been reported to
be depleted in an altered gut microbiome of patients with coronary
artery diseases (60). Some of the well-known members of
Oscillospiraceae (e.g., Faecalibacterium) aid in the production of
butyrate and have the capacity to metabolize glucuronate (61). Compared to CTL, RS groups significantly downregulated known
opportunistic harmful bacteria, including Desulfovibrio [PTB],
Eggerthella [BEP, CKP] and Enterococcus [BEP, CKP], which are
associated with inflammatory conditions in the gut (62). In addition, The ASVs of each treatment group retrieved from 16S rRNA
sequencing yielded alterations in their metabolic pathways compared
to CTL. The upregulation of genes involved in lipid and carbohydrate
metabolism by chickpea RS had been demonstrated, which are
consistent with our study (65). The underrepresentation of genes
involved in carbohydrate (glycolysis/gluconeogenesis and galactose)
and amino acid metabolism (D-alanine and D-glutamate) by RS
consumption has been previously reported (66). In addition, these 11 10.3389/fnut.2023.1106463 10.3389/fnut.2023.1106463 Kadyan et al. more pronounced effect on males than females (85). In line with this,
we observed reductions in the gut permeability via FITC-dextran
assay among different RS and INU groups, which could be attributed
to increased expression of tight-junction proteins such as CLDN1,
CLDN4 and OCCL in the intestinal tissues. Lower expression of
claudins is associated with a hyper-permeable mucosal environment
and is correlated with the onset of type-1 diabetes (86). Although CKP
did not exhibit any lowering effect on gut permeability, it did enhance
the expression of some barrier-forming genes (CLDN1 and CLDN4),
which could likely be due to the reason that CKP induced highest
bacterial diversity with upregulation of many beneficial genera in
beside some other selected genera namely, Tyzzerella and Oscillibacter,
which have earlier been associated with obesity and impaired gut
permeability (3). Though we have not analyzed fecal butyrate levels,
increased gut permeability in males could also be attributed butyrate-
dependent faster turnover of intestinal cells via colonocyte
differentiation and maturation (55). Further experiments focusing on
the fecal concentration of SCFAs would provide more evidence on the
beneficial effects of RS on host health. 3.5. Resistant starches from different dietary
pulses differentially influence inflammatory
markers in males and females studies have reported a positive association of Alistipes with former
metabolism and a negative association of Parasutterella with the latter,
which were also evident in our study. The histidine metabolism
pathway, which was only affected by BEP seems to play a role in body
fat reduction and lower levels of systemic inflammation (76). Upregulation of the sphingolipid signaling pathway further emphasizes
diet-induced restructuring of the gut microbiome towards improved
metabolic function (77). However, it may be noted that the functional
prediction using PICRUSt based on 16 s rRNA gene may not
be exhaustive as ASVs are annotated using only about 300 bp amplicon
read length and thus may have limited application to predict actual
complex functional events. Nevertheless, it provides novel hypotheses
and future research direction to gain deeper insights on the functional
consequences of RS on gut and host health using comprehensive and
inclusive approaches such as whole genome metagenomics
and metatranscriptomics. Intake of dietary fibers, including RS are widely reported to lower
post-prandial glucose response, improving insulin sensitivity, and
modulate energy metabolism (75). In the current study, the physiological
benefits of RS (mainly LEN and CKP) on faster glucose clearance during
oral meal administration were mostly observed in males during
HFD-feeding. The anti-obesity effects of native (RS2) and retrograded
(RS3) lentil starch revealed decreased blood glucose following 6-weeks
of HFD-feeding (14). The modulatory role of dietary chickpeas and their
RS in controlling post-prandial hyperglycemia, hyperinsulinemia and
hyperlipidemia has been reported earlier (65, 78). In line with our study,
sex-specific variations in glycemic responses to western-style diet have
also been reported in wild-type C57BL/6 mice with males demonstrating
a higher response to GTT than females (79). Higher glucose tolerance
of females than males could be associated with increased levels of
adiponectin and reduced oxidative stress conferred by estrogen (80),
which could be a plausible factor explaining no marginal change in
glycemic response of females post RS intervention. Insulin sensitivity
was also marginally influenced by CKP group in males in current study. Improvement in insulin sensitivity upon RS intervention has earlier
been found to be effective for men with no apparent benefit observed in
women (81). 3.5. Resistant starches from different dietary
pulses differentially influence inflammatory
markers in males and females Apart from gender effects, such RS-specific variation in the
glycemic response could be intimately linked to differences in the
structural characteristics of different pulse starches such as amylose
chain length, granule shape and size, porosity, B-polymorphic content,
degree of polymerization and amylose/amylopectin ratio in the
starch (82). if
Ample evidence suggests the crucial role of sexual dimorphism in
immune responses, with reported differences in basal level of pro-and
anti-inflammatory cytokines in males and females (55, 87). High IL-10
and low TNF-α expression, which was more prominent in LEN, CKP
and INU groups, respectively, further highlighted their potential for
improving aging-associated intestinal immunomodulation. A similar
trend in intestinal inflammation, as measured by mRNA expression, by
retrograded starch extracted from Arenga pinnata has previously been
reported in aged mice (88). IL-10 secreted by Tregs confers anti-
inflammatory effects by controlling the production of pro-inflammatory
cytokines like TNF-α while interacting with dendric cells and
macrophages. We find that concomitant to IL-10, IL-17 expression was
also enhanced by LEN. Innate production of IL-17 under homeostatic
conditions is primarily driven by intestinal γδ T-cells, whereas Th17 cells
control the adaptive immunity of IL-17. Its role in regulating
adipogenesis and glucose metabolism, along with strengthening TJ
proteins like claudins and occludins, has previously been established
(89). This further aligns with a plausible protective mechanism of LEN
in promoting post-prandial glucose tolerance and epithelial integrity. More importantly, its protective role against metabolic syndrome has
been demonstrated by limiting gut dysbiosis and LPS translocation (90). Further studies unraveling the role of specific T-cells in regulating IL-17
after RS treatment could provide more insights, as elevated levels of
IL-17 could also induce inflammation. Nevertheless, we observed
improvement in the intestinal integrity and inflammatory markers,
further suggesting the protective role of these pulses-derived RS in the
maintenance of normal glucose homeostasis under HFD-induced
metabolic dysfunction (12). Low-grade inflammation and gut hyperpermeability could further
fuel the pathogenesis of various metabolic diseases in the elderly (83). We have previously shown how HFD-induced perturbations in the gut
microbiome may exacerbate pro-inflammatory signals via microbe-
associated lipopolysaccharide (LPS) production, thereby stimulating
gut leakiness and endotoxemia (12). Overall, the current study showed
that the RS supplementation ameliorated intestinal dysfunctions in
both sexes; however, the regulatory effects of RS on tight-junction (TJ)
and inflammatory genes are sex-specific. frontiersin.org Frontiers in Nutrition References 1. Nagpal, R, Newman, TM, Wang, S, Jain, S, Lovato, JF, and Yadav, H. Obesity-linked
gut microbiome dysbiosis associated with derangements in gut permeability and
intestinal cellular homeostasis independent of diet. J Diabetes Res. (2018) 2018:1–9. doi: 10.1155/2018/3462092 1. Nagpal, R, Newman, TM, Wang, S, Jain, S, Lovato, JF, and Yadav, H. Obesity-linked
gut microbiome dysbiosis associated with derangements in gut permeability and
intestinal cellular homeostasis independent of diet. J Diabetes Res. (2018) 2018:1–9. doi: 10.1155/2018/3462092 9. Lutsiv, T, Weir, TL, McGinley, JN, Neil, ES, Wei, Y, and Thompson, HJ. Compositional
changes of the high-fat diet-induced gut microbiota upon consumption of common
pulses. Nutrients. (2021) 13:3992. doi: 10.3390/nu13113992 10. Liu, H, Zhang, M, Ma, Q, Tian, B, Nie, C, Chen, Z, et al. Health beneficial effects of
resistant starch on diabetes and obesity via regulation of gut microbiota: a review. Food Funct. (2020) 11:5749–67. doi: 10.1039/D0FO00855A 2. Nagpal, R, Kumar, M, Yadav, A, Hemalatha, R, Yadav, H, Marotta, F, et al. Gut
microbiota in health and disease: an overview focused on metabolic inflammation. Benefic Microbes. (2016) 7:181–94. doi: 10.3920/bm2015.0062 11. Sangokunle, OO, Sathe, SK, and Singh, P. Purified starches from 18 pulses have
markedly different morphology, oil absorption and water absorption capacities,
swelling power, and turbidity. Starch-Stärke. (2020) 72:2000022. doi: 10.1002/
star.202000022 3. Zhang, Y, Chen, L, Hu, M, Kim, JJ, Lin, R, Xu, J, et al. Dietary type 2 resistant starch
improves systemic inflammation and intestinal permeability by modulating
microbiota and metabolites in aged mice on high-fat diet. Aging. (2020) 12:9173–87. doi: 10.18632/aging.103187 12. Ahmadi, S, Nagpal, R, Wang, S, Gagliano, J, Kitzman, DW, Soleimanian-Zad, S, et al. Prebiotics from acorn and sago prevent high-fat-diet-induced insulin resistance via
microbiome–gut–brain axis modulation. J Nutr Biochem. (2019) 67:1–13. doi:
10.1016/j.jnutbio.2019.01.011 4. Nagpal, R, Shively, CA, Appt, SA, Register, TC, Michalson, KT, Vitolins, MZ, et al. Gut microbiome composition in non-human primates consuming a Western or
Mediterranean diet. Front Nutr. (2018) 5:28. doi: 10.3389/fnut.2018.00028 5. Kadyan, S, Sharma, A, Arjmandi, BH, Singh, P, and Nagpal, R. Prebiotic potential of
dietary beans and pulses and their resistant starch for aging-associated gut and
metabolic health. Nutrients. (2022) 14:1726. doi: 10.3390/nu14091726 13. Cui, W, Ma, Z, Li, X, and Hu, X. Structural rearrangement of native and processed
pea starches following simulated digestion in vitro and fermentation characteristics
of their resistant starch residues using human fecal inoculum. Int J Biol Macromol. (2021) 172:490–502. doi: 10.1016/j.ijbiomac.2021.01.092 6. Conflict of interest The authors declare that the research was conducted in the absence
of any commercial or financial relationships that could be construed as
a potential conflict of interest. Supplementary material The Supplementary material for this article can be found online
at:
https://www.frontiersin.org/articles/10.3389/fnut.2023.1106463/
full#supplementary-material Ethics statement This study was carried out in accordance with the recommendations
and guidelines of the Institutional Animal Care and Use Committee. The
protocol was approved by the Institutional Animal Care and Use
Committee at Florida State University (PROTO202100008). 3.5. Resistant starches from different dietary
pulses differentially influence inflammatory
markers in males and females The variation in the
expression of TJ genes in the intestine of males and females have been
reported in previous studies (84) reporting altered expression of
multiple TJ genes in the intestine of males and females using zonulin
transgenic mouse model and revealing sex-specific altered TJ genes
expression with reduction of CLDN15, CLDN5, JAM3, and Myo1C
genes only in males’ duodenum while downregulation of CLDN15,
CLDN7, and ZO2 in females’ colon. Earlier study using same model
demonstrated increased gut permeability in both genders with slightly In conclusion, our study provides novel insights pertaining to the
prebiotic potential of dietary pulses-derived RS in positively modulating
the gut microbiome architecture, strengthening the intestinal epithelial
barrier integrity, and attenuating inflammation in a preclinical model of
human microbiome and aging. Our findings also demonstrate that the
prebiotic effects of RS differ among host sexes, further emphasizing the
role of sexual dimorphism at microbiome, immune and metabolic level,
which is crucial for establishing universality of health claims imparted
by functional foods. However, the studied effects are specific to RS from
specific beans and pulses, which is intelligible considering the structural
differences associated with different pulse varieties. Nonetheless, based
on the evidence accumulated from this study, LEN and CKP seemed to 12 Kadyan et al. 10.3389/fnut.2023.1106463 10.3389/fnut.2023.1106463 Acknowledgments The authors wish to thank all the lab members, the Institutional
Animal Care and Use Committees members, and fellow colleagues for
their help and cooperation in these studies. Publisher’s note All claims expressed in this article are solely those of the authors and
do not necessarily represent those of their affiliated organizations, or
those of the publisher, the editors and the reviewers. Any product that
may be evaluated in this article, or claim that may be made by its
manufacturer, is not guaranteed or endorsed by the publisher. Data availability statement The original contributions presented in the study are publicly
available. This data can be found at: https://ncbi.nlm.nih.gov/bioproject/
PRJNA902407. Funding deliver more favorable outcomes in modulating gut microbiome and
improving metabolic health. Still, further studies unraveling the changes
in global microbial metabolomic, and transcriptomic arrays would be of
great interest and importance in ascertaining the direct functional
consequences of RS on host health. Future efforts will also be directed
towards understanding the microstructural differences in RS to precisely
target
sexually
dimorphic
gut
microbiome
for
preventing
metabolic disorders. This work was supported by funding from the Pulse Crop Health
Initiative program of the United States Department of Agriculture
(USDA-ARS) to RN. Author contributions SK performed experiments, analyzed data, and wrote manuscript. GP analyzed data and assisted in manuscript drafting. PS assisted in
starch extractions, revised, and edited manuscript. BA provided critical
inputs and advices on experimental design, data interpretation,
reviewed, and edited manuscript. RN conceived the idea, supervised the
study, analyzed and interpretated data, revised, and edited the
manuscript. All authors reviewed and approved the final version of
the manuscript. References FEMS
Microbiol
Ecol. (2013)
83:299–309. doi:
10.1111/j.1574-6941.2012.01475.x 19. Ahmadi, S, Razazan, A, Nagpal, R, Jain, S, Wang, B, Mishra, SP, et al. Metformin
reduces aging-related leaky gut and improves cognitive function by beneficially
modulating gut microbiome/goblet cell/mucin axis. J Gerontol Ser A. (2020) 75:e9–
e21. doi: 10.1093/gerona/glaa056 44. Barouei, J, Bendiks, Z, Martinic, A, Mishchuk, D, Heeney, D, Hsieh, Y, et al. Microbiota, metabolome, and immune alterations in obese mice fed a high-fat diet
containing type 2 resistant starch. Mol Nutr Food Res. (2017) 61:1700184. doi:
10.1002/mnfr.201700184 20. Singh, V, San Yeoh, B, Chassaing, B, Xiao, X, Saha, P, Olvera, RA, et al. Dysregulated
microbial fermentation of soluble fiber induces cholestatic liver cancer. Cells. (2018)
175:679–694.e22. doi: 10.1016/j.cell.2018.09.004 45. Belobrajdic, DP, King, RA, Christophersen, CT, and Bird, AR. Dietary resistant starch
dose-dependently reduces adiposity in obesity-prone and obesity-resistant male rats. Nutr Metab. (2012) 9:93–10. doi: 10.1186/1743-7075-9-93 21. Vijay-Kumar, M, Aitken, JD, Carvalho, FA, Cullender, TC, Mwangi, S, Srinivasan, S,
et al. Metabolic syndrome and altered gut microbiota in mice lacking toll-like
receptor 5. Science. (2010) 328:228–31. doi: 10.1126/science.1179721 46. Silvi, S, Rumney, C, Cresci, A, and Rowland, I. Resistant starch modifies gut
microflora and microbial metabolism in human flora-associated rats inoculated with
faeces from Italian and UK donors. J Appl Microbiol. (1999) 86:521–30. doi:
10.1046/j.1365-2672.1999.00696.x 22. Sangokunle, OO. Exploration of Purified Pulse Starches for Food and Health. [Dissertation]. Tallahassee: Florida State University (2021). 47. Jakobsdottir, G, Jädert, C, Holm, L, and Nyman, ME. Propionic and butyric acids,
formed in the caecum of rats fed highly fermentable dietary fibre, are reflected in
portal and aortic serum. Br J Nutr. (2013) 110:1565–72. doi: 10.1017/
S0007114513000809 23. Nagpal, R, Indugu, N, and Singh, P. Distinct gut microbiota signatures in mice treated
with commonly used food preservatives. Microorganisms. (2021) 9:2311. doi:
10.3390/microorganisms9112311 24. Clark, M, Centner, AM, Ukhanov, V, Nagpal, R, and Salazar, G. Gallic acid
ameliorates atherosclerosis and vascular senescence and remodels the microbiome
in a sex-dependent manner in ApoE−/− mice. J Nutr Biochem. (2022) 110:109132. doi: 10.1016/j.jnutbio.2022.109132 48. Bird, AR, Brown, IL, and Topping, DL. Starches, resistant starches, the gut microflora
and human health. Curr Issues Intest Microbiol. (2000) 1:25–37. PMID: 11709851 49. Jiminez, JA, Uwiera, TC, Abbott, DW, Uwiera, RR, and Inglis, GD. Impacts of
resistant starch and wheat bran consumption on enteric inflammation in relation to
colonic bacterial community structures and short-chain fatty acid concentrations in
mice. Gut Pathog. References Darmadi-Blackberry, I, Wahlqvist, ML, Kouris-Blazos, A, Steen, B, Lukito, W, Horie, Y,
et al. Legumes: the most important dietary predictor of survival in older people of
different ethnicities. Asia Pac J Clin Nutr. (2004) 13:217–20. PMID: 15228991 14. Xu, J, Ma, Z, Li, X, Liu, L, and Hu, X. A more pronounced effect of type III resistant
starch vs. type II resistant starch on ameliorating hyperlipidemia in high fat diet-fed
mice is associated with its supramolecular structural characteristics. Food Funct. (2020) 11:1982–95. doi: 10.1039/C9FO02025J 7. Mitchell, DC, Marinangeli, CP, Pigat, S, Bompola, F, Campbell, J, Pan, Y, et al. Pulse
intake improves nutrient density among US adult consumers. Nutrients. (2021)
13:2668. doi: 10.3390/nu13082668 15. Zhou, D, Ma, Z, and Hu, X. Isolated pea resistant starch substrates with different
structural features modulate the production of short-chain fatty acids and
metabolism of microbiota in anaerobic fermentation in vitro. J Agric Food Chem. (2021) 69:5392–404. doi: 10.1021/acs.jafc.0c08197 8. Marinangeli, C, Harding, S, Zafron, M, and Rideout, T. A systematic review of the
effect of dietary pulses on microbial populations inhabiting the human gut. Benef
Microbes. (2020) 11:457–68. doi: 10.3920/BM2020.0028 13 Frontiers in Nutrition frontiersin.org frontiersin.org Kadyan et al. 10.3389/fnut.2023.1106463 16. Park, JC, and Im, S-H. Of men in mice: the development and application of a
humanized gnotobiotic mouse model for microbiome therapeutics. Exp Mol Med. (2020) 52:1383–96. doi: 10.1038/s12276-020-0473-2 41. Heberle, H, Meirelles, GV, da Silva, FR, Telles, GP, and Minghim, R. InteractiVenn:
a web-based tool for the analysis of sets through Venn diagrams. BMC Bioinformat. (2015) 16:1–7. doi: 10.1186/s12859-015-0611-3 17. Chen, Y, McGee, R, Vandemark, G, Brick, M, and Thompson, HJ. Dietary fiber
analysis of four pulses using AOAC 2011.25: implications for human health. Nutrients. (2016) 8:829. doi: 10.3390/nu8120829 42. Keenan, MJ, Janes, M, Robert, J, Martin, RJ, Raggio, AM, McCutcheon, KL, et al. Resistant starch from high amylose maize (HAM-RS2) reduces body fat and increases
gut bacteria in ovariectomized (OVX) rats. Obesity. (2013) 21:981–4. doi: 10.1002/
oby.20109 18. Tuncil, YE, Thakkar, RD, Arioglu-Tuncil, S, Hamaker, BR, and Lindemann, SR. Fecal
microbiota responses to bran particles are specific to cereal type and in vitro
digestion methods that mimic upper gastrointestinal tract passage. J Agric Food
Chem. (2018) 66:12580–93. doi: 10.1021/acs.jafc.8b03469 43. Sybille, T, June, Z, Michael, K, Roy, M, and Maria, LM. The intestinal microbiota in
aged mice is modulated by dietary resistant starch and correlated with improvements
in
host
responses. References (2016) 8:1–20. doi: 10.1186/s13099-016-0149-6 25. Munley, JA, Kelly, LS, Pons, EE, Kannan, KB, Coldwell, PS, Whitley, EM, et al. Multicompartmental traumatic injury and the microbiome: shift to a Pathobiome. J
Trauma Acute Care Surg. (2022) 94:15–22. doi: 10.1097/TA.0000000000003803 50. Martínez, I, Kim, J, Duffy, PR, Schlegel, VL, and Walter, J. Resistant starches types 2
and 4 have differential effects on the composition of the fecal microbiota in human. PLoS ONE. (2010) 5:e15046. doi: 10.1371/journal.pone.0015046 26. Saccon, TD, Nagpal, R, Yadav, H, Cavalcante, MB, Nunes, AD d C, Schneider, A, et al. Senolytic combination of Dasatinib and quercetin alleviates intestinal senescence and
inflammation and modulates the gut microbiome in aged mice. J Gerontol Ser A. (2021) 76:1895–905. doi: 10.1093/gerona/glab002 51. Vital, M, Howe, A, Bergeron, N, Krauss, RM, Jansson, JK, and Tiedje, JM. Metagenomic insights into the degradation of resistant starch by human gut
microbiota. Appl Environ Microbiol. (2018) 84:e01562–18. doi: 10.1128/
AEM.01562-18 27. Nagpal, R, Neth, BJ, Wang, S, Craft, S, and Yadav, H. Modified Mediterranean-
ketogenic diet modulates gut microbiome and short-chain fatty acids in association
with Alzheimer’s disease markers in subjects with mild cognitive impairment. EBioMedicine. (2019) 47:529–42. doi: 10.1016/j.ebiom.2019.08.032 52. Guan, N, He, X, Wang, S, Liu, F, Huang, Q, Fu, X, et al. Cell wall integrity of pulse
modulates the in vitro fecal fermentation rate and microbiota composition. J Agric
Food Chem. (2020) 68:1091–100. doi: 10.1021/acs.jafc.9b06094 28. Bolyen, E, Rideout, JR, Dillon, MR, Bokulich, NA, Abnet, CC, Al-Ghalith, GA, et al. Reproducible, interactive, scalable and extensible microbiome data science using
QIIME 2. Nat Biotechnol. (2019) 37:852–7. doi: 10.1038/s41587-019-0209-9 53. Org, E, Mehrabian, M, Parks, BW, Shipkova, P, Liu, X, Drake, TA, et al. Sex differences
and hormonal effects on gut microbiota composition in mice. Gut Microbes. (2016)
7:313–22. doi: 10.1080/19490976.2016.1203502 29. Callahan, BJ, McMurdie, PJ, Rosen, MJ, Han, AW, Johnson, AJA, and Holmes, SP. DADA2: high-resolution sample inference from Illumina amplicon data. Nat
Methods. (2016) 13:581–3. doi: 10.1038/nmeth.3869 54. Suzuki, Y, Ikeda, K, Sakuma, K, Kawai, S, Sawaki, K, Asahara, T, et al. Association
between yogurt consumption and intestinal microbiota in healthy young adults
differs by host gender. Front Microbiol. (2017) 8:847. doi: 10.3389/fmicb.2017.00847 30. Katoh, K, Misawa, K, Kuma, K, and Miyata, T. MAFFT: a novel method for rapid
multiple sequence alignment based on fast Fourier transform. Nucleic Acids Res. (2002) 30:3059–66. doi: 10.1093/nar/gkf436 55. References Shastri, P, McCarville, J, Kalmokoff, M, Brooks, SP, and Green-Johnson, JM. Sex
differences in gut fermentation and immune parameters in rats fed an oligofructose-
supplemented diet. Biol Sex Differ. (2015) 6:1–12. doi: 10.1186/s13293-015-0031-0 31. Bokulich, NA, Kaehler, BD, Rideout, JR, Dillon, M, Bolyen, E, Knight, R, et al. Optimizing taxonomic classification of marker-gene amplicon sequences with
QIIME 2’s q2-feature-classifier plugin. Microbiome. (2018) 6:1–17. doi: 10.1186/
s40168-018-0470-z 56. Ke, X, Walker, A, Haange, S-B, Lagkouvardos, I, Liu, Y, Schmitt-Kopplin, P, et al. Synbiotic-driven improvement of metabolic disturbances is associated with changes
in the gut microbiome in diet-induced obese mice. Mol Metab. (2019) 22:96–109. doi: 10.1016/j.molmet.2019.01.012 32. Robeson, M, O’Rourke, D, Kaehler, B, Ziemski, M, Dillon, M, Foster, J, et al. RESCRIPt: reproducible sequence taxonomy reference database management for the
masses. bioRxiv. (2020). doi: 10.1101/2020.10.05.326504 57. Daillère, R, Vétizou, M, Waldschmitt, N, Yamazaki, T, Isnard, C, Poirier-Colame, V,
et
al. Enterococcus
hirae
and
Barnesiella
intestinihominis
facilitate
cyclophosphamide-induced therapeutic immunomodulatory effects. Immunity. (2016) 45:931–43. doi: 10.1016/j.immuni.2016.09.009 33. Miller, B, Mainali, R, Nagpal, R, and Yadav, H. A newly developed Synbiotic yogurt
prevents diabetes by improving the microbiome–intestine–pancreas Axis. Int J Mol
Sci. (2021) 22:1647. doi: 10.3390/ijms22041647 58. Lei, S, Liu, L, Ding, L, Zhang, Y, and Zeng, H. Lotus seed resistant starch affects the
conversion of sodium taurocholate by regulating the intestinal microbiota. Int J Biol
Macromol. (2021) 186:227–36. doi: 10.1016/j.ijbiomac.2021.07.031 34. Kruskal, WH, and Wallis, WA. Use of ranks in one-criterion variance analysis. J
Am Stat Assoc. (1952) 47:583–621. doi: 10.1080/01621459.1952.10483441 35. Anderson, MJ. A new method for non-parametric multivariate analysis of variance. Austral Ecol. (2001) 26:32–46. doi: 10.1111/j.1442-9993.2001.01070.pp.x 59. Parada Venegas, D, De la Fuente, MK, Landskron, G, González, MJ, Quera, R,
Dijkstra, G, et al. Short chain fatty acids (SCFAs)-mediated gut epithelial and
immune regulation and its relevance for inflammatory bowel diseases. Front
Immunol. (2019) 10:277. doi: 10.3389/fimmu.2019.00277 36. Breiman,
L. Random
forests. Mach
Learn. (2001)
45:5–32. doi:
10.1023/A:1010933404324 37. Langille, MG, Zaneveld, J, Caporaso, JG, McDonald, D, Knights, D, Reyes, JA, et al. Predictive functional profiling of microbial communities using 16S rRNA marker
gene sequences. Nat Biotechnol. (2013) 31:814–21. doi: 10.1038/nbt.2676 60. Toya, T, Corban, MT, Marrietta, E, Horwath, IE, Lerman, LO, Murray, JA, et al. Coronary artery disease is associated with an altered gut microbiome composition. PLoS One. (2020) 15:e0227147. doi: 10.1371/journal.pone.0227147 38. Douglas, GM, Maffei, VJ, Zaneveld, JR, Yurgel, SN, Brown, JR, Taylor, CM, et al. References PICRUSt2 for prediction of metagenome functions. Nat Biotechnol. (2020) 38:685–8. doi: 10.1038/s41587-020-0548-6 61. Gophna, U, Konikoff, T, and Nielsen, HB. Oscillospira and related bacteria–from
metagenomic species to metabolic features. Environ Microbiol. (2017) 19:835–41. doi:
10.1111/1462-2920.13658 39. Segata, N, Izard, J, Waldron, L, Gevers, D, Miropolsky, L, Garrett, WS, et al. Metagenomic biomarker discovery and explanation. Genome Biol. (2011) 12:1–18. doi: 10.1186/gb-2011-12-6-r60 62. Warman, DJ, Jia, H, and Kato, H. The potential roles of probiotics, resistant starch,
and resistant proteins in ameliorating inflammation during aging (Inflammaging). Nutrients. (2022) 14:747. doi: 10.3390/nu14040747 63. Zhou, Y, Zhao, S, Jiang, Y, Wei, Y, and Zhou, X. Regulatory function of buckwheat-
resistant starch supplementation on lipid profile and gut microbiota in mice fed with
a high-fat diet. J Food Sci. (2019) 84:2674–81. doi: 10.1111/1750-3841.14747 40. Parks, DH, Tyson, GW, Hugenholtz, P, and Beiko, RG. STAMP: statistical analysis of
taxonomic and functional profiles. Bioinformatics. (2014) 30:3123–4. doi: 10.1093/
bioinformatics/btu494 14 Frontiers in Nutrition frontiersin.org frontiersin.org frontiersin.org Kadyan et al. 10.3389/fnut.2023.1106463 64. Wan, J, Wu, Y, Pham, Q, Li, RW, Yu, L, Chen, M-H, et al. Effects of differences in
resistant starch content of rice on intestinal microbial composition. J Agric Food
Chem. (2021) 69:8017–27. doi: 10.1021/acs.jafc.0c07887 gut microbiota. Genom. Proteom. Bioinformat. (2019) 17:64–75. doi: 10.1016/j. gpb.2019.03.001 78. Yang, Y, Zhou, L, Gu, Y, Zhang, Y, Tang, J, Li, F, et al. Dietary chickpeas reverse
visceral adiposity, dyslipidaemia and insulin resistance in rats induced by a chronic
high-fat diet. Br J Nutr. (2007) 98:720–6. doi: 10.1017/S0007114507750870 65. Zhao, M, Cui, W, Hu, X, and Ma, Z. Anti-hyperlipidemic and ameliorative effects of
chickpea starch and resistant starch in mice with high fat diet induced obesity are
associated with their multi-scale structural characteristics. Food Funct. (2022)
13:5135–52. doi: 10.1039/D1FO04354D 79. Sheng, L, Jena, PK, Liu, H-X, Kalanetra, KM, Gonzalez, FJ, French, SW, et al. Gender
differences in bile acids and microbiota in relationship with gender dissimilarity in
steatosis induced by diet and FXR inactivation. Sci Rep. (2017) 7:1–12. doi: 10.1038/
s41598-017-01576-9 66. Zhang, L, Ouyang, Y, Li, H, Shen, L, Ni, Y, Fang, Q, et al. Metabolic phenotypes and
the gut microbiota in response to dietary resistant starch type 2 in normal-weight
subjects: a randomized crossover trial. Sci Rep. (2019) 9:1–11. doi: 10.1038/
s41598-018-38216-9 80. Ahmed, S, and Spence, JD. Sex differences in the intestinal microbiome: interactions
with risk factors for atherosclerosis and cardiovascular disease. Biol Sex Differ. References (2021)
12:1–12. doi: 10.1186/s13293-021-00378-z 67. Jian, C, Silvestre, MP, Middleton, D, Korpela, K, Jalo, E, Broderick, D, et al. Gut
microbiota predicts body fat change following a low-energy diet: a PREVIEW
intervention study. Genome Med. (2022) 14:1–18. doi: 10.1186/s13073-022-01053-7 81. Maki, KC, Pelkman, CL, Finocchiaro, ET, Kelley, KM, Lawless, AL, Schild, AL, et al. Resistant starch from high-amylose maize increases insulin sensitivity in overweight
and obese men. J Nutr. (2012) 142:717–23. doi: 10.3945/jn.111.152975 68. Nakai, M, Ribeiro, RV, Stevens, BR, Gill, P, Muralitharan, RR, Yiallourou, S, et al. Essential hypertension is associated with changes in gut microbial metabolic
pathways: a multisite analysis of ambulatory blood pressure. Hypertension. (2021)
78:804–15. doi: 10.1161/HYPERTENSIONAHA.121.17288 82. Parada, J, and Aguilera, J. Starch matrices and the glycemic response. Food Sci Technol
Int. (2011) 17:187–204. doi: 10.1177/1082013210387712 83. Chung, HY, Kim, DH, Lee, EK, Chung, KW, Chung, S, Lee, B, et al. Redefining
chronic inflammation in aging and age-related diseases: proposal of the
senoinflammation concept. Aging Dis. (2019) 10:367–82. doi: 10.14336/AD.2018.0324 69. Yue, C, Li, M, Li, J, Han, X, Zhu, H, Yu, G, et al. Medium-, long-and medium-chain-
type structured lipids ameliorate high-fat diet-induced atherosclerosis by regulating
inflammation, adipogenesis, and gut microbiota in ApoE−/− mice. Food Funct. (2020) 11:5142–55. doi: 10.1039/D0FO01006E 84. Miranda-Ribera, A, Ennamorati, M, Serena, G, Cetinbas, M, Lan, J, Sadreyev, RI,
et al. Exploiting the zonulin mouse model to establish the role of primary impaired
gut barrier function on microbiota composition and immune profiles. Front
Immunol. (2019) 10:2233. doi: 10.3389/fimmu.2019.02233 70. Zhou, Y, Wei, Y, Yan, B, Zhao, S, and Zhou, X. Regulation of tartary buckwheat-
resistant starch on intestinal microflora in mice fed with high-fat diet. Food Sci Nutr. (2020) 8:3243–51. doi: 10.1002/fsn3.1601 85. Sturgeon, C, Lan, J, and Fasano, A. Zonulin transgenic mice show altered gut
permeability and increased morbidity/mortality in the DSS colitis model. Ann N Y
Acad Sci. (2017) 1397:130–42. doi: 10.1111/nyas.13343 71. Senevirathne, RN, Janes, M, Keenan, MJ, Martin, RJ, Raggio, AM, McCutcheon, KL,
et al. Dietary resistant starch increases Bifidobacterium spp./lactobacillus spp. and
clostridia spp. in the gut of mice fed low and moderate fat diets. FASEB J. (2009)
23:719–6. doi: 10.1096/fasebj.23.1_supplement.719.6 86. Neu, J, Reverte, CM, Mackey, AD, Liboni, K, Tuhacek-Tenace, LM, Hatch, M, et al. Changes in intestinal morphology and permeability in the biobreeding rat before the
onset of type 1 diabetes. J Pediatr Gastroenterol Nutr. (2005) 40:589–95. doi:
10.1097/01.MPG.0000159636.19346.C1 72. References DeMartino, P, and Cockburn, DW. Resistant starch: impact on the gut microbiome
and health. Curr Opin Biotechnol. (2020) 61:66–71. doi: 10.1016/j.copbio.2019. 10.008 87. Sankaran-Walters, S, Macal, M, Grishina, I, Nagy, L, Goulart, L, Coolidge, K, et al. Sex differences matter in the gut: effect on mucosal immune activation and
inflammation. Biol Sex Differ. (2013) 4:10. doi: 10.1186/2042-6410-4-10 73. Dobranowski, PA, and Stintzi, A. Resistant starch, microbiome, and precision
modulation. Gut Microbes. (2021) 13:1926842. doi: 10.1080/19490976.2021.1926842 74. Wen, J-J, Li, M-Z, Hu, J-L, Tan, H-Z, and Nie, S-P. Resistant starches and gut
microbiota. Food Chem. (2022) 387:132895. doi: 10.1016/j.foodchem.2022.132895 88. Ren, M, Li, M-Y, Lu, L-Q, Liu, Y-S, An, F-K, Huang, K, et al. Arenga pinnata resistant
starch modulate gut microbiota and ameliorate intestinal inflammation in aged mice. Nutrients. (2022) 14:3931. doi: 10.3390/nu14193931 75. Zhang, C, Qiu, M, Wang, T, Luo, L, Xu, W, Wu, J, et al. Preparation, structure
characterization, and specific gut microbiota properties related to anti-hyperlipidemic
action of type 3 resistant starch from Canna edulis. Food Chem. (2021) 351:129340. doi: 10.1016/j.foodchem.2021.129340 89. Zúñiga, LA, Shen, W-J, Joyce-Shaikh, B, Pyatnova, EA, Richards, AG, Thom, C, et al. IL-17 regulates adipogenesis, glucose homeostasis, and obesity. J Immunol. (2010)
185:6947–59. doi: 10.4049/jimmunol.1001269 76. DiNicolantonio, JJ, McCarty, MF, and OKeefe, JH. Role of dietary histidine in the
prevention of obesity and metabolic syndrome. Open Heart. (2018) 5:e000676. doi:
10.1136/openhrt-2017-000676 90. Pérez, MM, Martins, LM, Dias, MS, Pereira, CA, Leite, JA, Gonçalves, EC, et al. Interleukin-17/interleukin-17 receptor axis elicits intestinal neutrophil migration,
restrains gut dysbiosis and lipopolysaccharide translocation in high-fat diet-induced
metabolic syndrome model. Immunology. (2019) 156:339–55. doi: 10.1111/
imm.13028 77. Song, X, Zhong, L, Lyu, N, Liu, F, Li, B, Hao, Y, et al. Inulin can alleviate metabolism
disorders in Ob/Ob mice by partially restoring leptin-related pathways mediated by 15 frontiersin.org 15 Frontiers in Nutrition Frontiers in Nutrition
|
https://openalex.org/W4294169472
|
https://www.frontiersin.org/articles/10.3389/fnut.2022.986731/pdf
|
English
| null |
The change in nutritional status is related to cardiovascular events in patients with pacemaker implantation: A 4-year follow-up study
|
Frontiers in nutrition
| 2,022
|
cc-by
| 6,158
|
OPEN ACCESS OPEN ACCESS
EDITED BY
Lilia Castillo-Martinez,
Instituto Nacional de Ciencias Médicas
y Nutrición Salvador Zubirán
(INCMNSZ), Mexico
REVIEWED BY
Dulce González-Islas,
Instituto Nacional de Enfermedades
Respiratorias-México (INER), Mexico
José L. Villanueva-Juárez,
Instituto Nacional de Ciencias Médicas
y Nutrición Salvador Zubirán
(INCMNSZ), Mexico
*CORRESPONDENCE
Chen Chen
chenchenowen@126.com
Xiang Hu
huxiang@wmu.edu.cn
†These authors have contributed
equally to this work
SPECIALTY SECTION
This article was submitted to
Clinical Nutrition,
a section of the journal
Frontiers in Nutrition
RECEIVED 05 July 2022
ACCEPTED 09 August 2022
PUBLISHED 02 September 2022
CITATION
Wang K, Lian L, Chen C, Wang M,
Chen C and Hu X (2022) The change
in nutritional status is related
to cardiovascular events in patients
with pacemaker implantation: A 4-year
follow-up study. Front. Nutr. 9:986731. doi: 10.3389/fnut.2022.986731
COPYRIGHT
© 2022 Wang, Lian, Chen, Wang, Chen
and Hu. This is an open-access article
distributed under the terms of the
Creative Commons Attribution License
(CC BY). The use, distribution or
reproduction in other forums is
permitted, provided the original
author(s) and the copyright owner(s)
are credited and that the original
publication in this journal is cited, in
accordance with accepted academic
practice. No use, distribution or
reproduction is permitted which does
not comply with these terms. EDITED BY
Lilia Castillo-Martinez,
Instituto Nacional de Ciencias Médicas
y Nutrición Salvador Zubirán
(INCMNSZ), Mexico Kaijing Wang1†, Liyou Lian2,3†, Chengpu Chen2,3,
Meiling Wang2,3, Chen Chen2,3* and Xiang Hu4* 1Department of Ultrasound, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou,
China, 2Department of Cardiology, The First Affiliated Hospital of Wenzhou Medical University,
Wenzhou, China, 3The Key Lab of Cardiovascular Disease, Science and Technology of Wenzhou,
Wenzhou, China, 4Key Laboratory of Clinical Laboratory Diagnosis and Translational Research
of Zhejiang Province, Department of Endocrine and Metabolic Diseases, The First Affiliated Hospital
of Wenzhou Medical University, Wenzhou, China Background: The aim of our study was to evaluate changes in nutritional
status as measured by the prognostic nutritional index (PNI) and geriatric
nutritional risk index (GNRI) scores, and their abilities to predict clinical
prognosis in patients with pacemaker implantation (PMI). Methods: A total of 595 patients who underwent permanent PMI from January
2011 to December 2020 were included. PNI and GNRI scores were separately
calculated at the beginning day of PMI operation and at the end of 12-
month follow-up, and their net changes (1) were calculated by PNI or
GNRI scores at follow-up minus the corresponding scores on admission. TYPE Original Research
PUBLISHED 02 September 2022
DOI 10.3389/fnut.2022.986731 TYPE Original Research
PUBLISHED 02 September 2022
DOI 10.3389/fnut.2022.986731 TYPE Original Research
PUBLISHED 02 September 2022
DOI 10.3389/fnut.2022.986731 COPYRIGHT
© 2022 Wang, Lian, Chen, Wang, Chen
and Hu. This is an open-access article
distributed under the terms of the
Creative Commons Attribution License
(CC BY). The use, distribution or
reproduction in other forums is
permitted, provided the original
author(s) and the copyright owner(s)
are credited and that the original
publication in this journal is cited, in
accordance with accepted academic
practice. No use, distribution or
reproduction is permitted which does
not comply with these terms. OPEN ACCESS The
cohort patients were divided into low risk of malnutritional status (1PNI or
1GNRI scores ≥0) and high risk of malnutritional status (1PNI or 1GNRI
scores < 0) groups. Primary outcome measure was a composite major
adverse cardiovascular event (MCE), defined as heart failure hospitalization
(HFH), myocardial infarction (MI), stroke, or death from any cause, presented
as hazard ratios (HR) with 95% confidence intervals (CI) calculated by MCE in
the crude or multivariate-adjusted Cox Proportional Hazards models. Receiver
operating characteristic (ROC) curve analysis was used to compare the
differential ability to predict incident MCEs between1PNI and1GNRI scores. C
O
Wang K, Lian L, Chen C, Wang M,
Chen C and Hu X (2022) The change
in nutritional status is related
to cardiovascular events in patients
with pacemaker implantation: A 4-year
follow-up study. Front Nutr 9:986731 COPYRIGHT
© 2022 Wang, Lian, Chen, Wang, Chen
and Hu. This is an open-access article
distributed under the terms of the
Creative Commons Attribution License
(CC BY). The use, distribution or
reproduction in other forums is
permitted, provided the original
author(s) and the copyright owner(s)
are credited and that the original
publication in this journal is cited, in
accordance with accepted academic
practice. No use, distribution or
reproduction is permitted which does
not comply with these terms. Results: In total, 16% of patients developed the MCE during the follow-
up. The cumulative event rates determined by Kaplan–Meier analysis were
significantly higher in the high risk of malnutritional patients compared to the
low risk of malnutritional patients (P < 0.05). Adjusted multivariate analysis
showed that decreased PNI scores (HR: 2.228, 95% CI: 1.482–3.350) and
decreased GNRI scores (HR: 2.178, 95% CI: 1.439–3.295) were independently
associated with favorable outcomes. ROC curve analysis revealed an area Frontiers in Nutrition 01 frontiersin.org Wang et al. 10.3389/fnut.2022.986731 10.3389/fnut.2022.986731 under curve (AUC) of 0.586 for1PNI scores and AUC of 0.592 for 1GNRI
scores, but their predictive abilities were not statistically different. Conclusion: Either positive change of PNI or GNRI scores were associated
with reduced risk of MCEs in patients with PMI, and they have similar
ability to predict clinical cardiometabolic risk. Additional enhancing nutritional
status during follow-up may help to prevent unfavorable prognosis in
clinical practices. pacemaker implantation, prognostic nutritional index, geriatric nutritional risk index,
nutritional status, prognosis constantly change; therefore, the prognostic value of PNI and
GNRI scores for adverse clinical outcomes could also change
over time. OPEN ACCESS A recent study (13) reported that change in PNI
score was associated with clinical outcomes in patients with
cardiac resynchronization therapy (CRT)-device. Another study
(14) proved that remaining at nutritional risk in transcatheter
aortic valve replacement group resulted in an increased risk
of mortality and HFH. Thus, positive changes in the PNI
and GNRI scores during follow-up following PMI may be
associated with improved cardiac function and subsequent
clinical outcomes; however, few studies regarding associations
of changes in nutritional status and clinical outcomes in patients
with pacemakers have been reported. Therefore, present study
evaluated changes in nutritional status as measured by the PNI
and GNRI scores, and their abilities to predict clinical prognosis
in patients with PMI. Introduction Pacemaker implantation (PMI) is an effective treatment for
patients who continue to have symptoms of heart failure (HF)
while receiving adequate medical care (1). It is well-established
that PMI can help reduce the rate of HF hospitalization
(HFH) and atrial fibrillation (AF) in pacing-dependent groups
(2). However, some individuals are unable to benefit from
this therapy for a variety of reasons, resulting in suboptimal
outcomes (3). Malnutrition
is
a
common
comorbidity
in
elderly
individuals and is related with a worse prognosis (4, 5). Additionally, PMI recipients have grown older and more
complex,
with
numerous
comorbidities
(6). Thus,
the
nutritional status in this group needs additional monitoring. Several studies have demonstrated an association between a
single nutritional indicator (7–9), such as albumin, and poor
outcomes in individuals with chronic HF. However, to evaluate
the nutritional status, more complex and objective indices
are needed because single indicator might be inflected by
many external factors. Take albumin for instance. Albumin
(and other serum proteins generated by the liver) is altered
not just by nutritional state, but also by inflammation and
infection, limiting its utility in acutely unwell individuals. Additionally, albumin’s extended half-life reduces its utility for
monitoring short-term changes in both calorie and protein
intakes. Several nutritional screening measures have been
presented for the purpose of assessing malnutrition and its
long-term prognosis (10). Frontiers in Nutrition pacemaker implantation, prognostic nutritional index, geriatric nutritional risk index,
nutritional status, prognosis Study subjects This was a retrospective analysis of patients who had
successful PMI at the First Affiliated Hospital of Wenzhou
Medical University between January 2011 and December
2020 with complete follow-up data. Blood collection, 12-lead
electrocardiography (ECG), and echocardiography were used
to evaluate the clinical status prior to PMI. The indications
for pacemakers were recorded [atrioventricular block (AVB),
sick sinus syndrome (SSS), and atrial fibrillation (AF) with
bradycardia, left bundle branch block (LBBB) with HF, AF
with atrioventricular node ablation (ANVA)]. All patients were
evaluated thoroughly 1 year after implantation, including blood
sample analysis and echocardiography. Patients who were
lost to follow-up or lacked the necessary data were omitted
from this study. The prognostic nutritional index (PNI) score is generated
using blood albumin and lymphocyte counts to describe
the immunological nutritional status of surgical patients and
to estimate the probability of developing a complication
(11). Another nutrition screening measure frequently used
in individuals with HF is the geriatric nutritional risk index
(GNRI) score (12). These two indices can be determined using
only simple blood biomarkers. The nutritional status might Frontiers in Nutrition 02 frontiersin.org Wang et al. Wang et al. 10.3389/fnut.2022.986731 10.3389/fnut.2022.986731 99th percentile upper reference limit) with necrosis in a
clinical situation consistent with myocardial ischemia (15). The status and/or dates of death of all patients were gathered
from the patients’ medical records or attending physicians
at the patient’s referring hospital. All patients were followed
in this study. The duration of survival was calculated
from the date of PMI to the date of readmission, death,
or last follow-up. The net changes (1) were calculated by PNI or GNRI scores
at follow-up minus the corresponding scores on admission. The
cohort patients were divided into low risk of malnutritional
status (1PNI or 1GNRI scores ≥0) and high risk of
malnutritional status (1PNI or 1GNRI scores < 0) groups. We investigated the influence of 1PNI and 1GNR scores
following PMI on laboratory data, cardiac function, and
significant adverse cardiovascular events (MCEs). The PNI and GNRI scores are nutritional risk indicators that
are associated with the severity of malnutrition and mortality
rate in hospitalized patients. They were calculated as following
formula: Data collection Baseline information was investigated, including BMI,
smoking and drinking history and concurrent disease. The
echocardiographic parameters included the left ventricular
end-diastolic diameter (LVEDD), left atrial diameters (LAD),
LV ejection fraction (LVEF), mitral regurgitation (MR),
and tricuspid regurgitation (TR). Laboratory data included
hemoglobin, total lymphocyte count, albumin, triglycerides
(TG), total cholesterol (TC), high density lipoprotein (HDL),
low density lipoprotein (LDL), aspartate aminotransferase
(AST), white blood cell (WBC), hemoglobin (Hb), platelets
(PLT), and the PNI and GNRI scores. Statistical analyses Data were presented as percentages for categorical variables
and mean ± standard deviation (SD) or median and
interquartile range (IQR) for continuous variables. Group
differences were evaluated using Student t-tests or Mann–
Whitney U tests for continuous variables and chi-square
or Fisher exact tests for categorical variables. The crude
or multivariate-adjusted Cox Proportional Hazards models
were used to estimate the hazard ratios (HR) and 95%
confidence intervals (CI) calculated by MCE without or with the
additional adjustments including baseline sociodemographic
parameters, lifestyle factors, prevalent diseases, with the low
risk of malnutrition group as a reference category. The
cumulative incidence curve of adverse events was plotted
via the Kaplan-Meier method, with statistical significance
examined by the log-rank test. Receiver operating characteristic
(ROC) curve analysis was used to compare the difference
of the changes in PNI and GNRI scores in the ability of
predicting prognosis. ROC curves were generated for the
changes of PNI and GNRI scores, and areas under the
curve (AUCs), cut-of values, sensitivities, and specificities were
calculated. All analyses were performed using R V.4.1.3 or IBM
SPSS version 26.0 (IBM Corp., Armonk, NY, United States)
and tests were two-sided and P < 0.05 was considered
statistically significant. PNI scores = serum albumin (g/L) + 0.005 × total
lymphocyte count (per mm3) GNRI
scores
=
1.489
×
serum
albumin
level
(g/L) + 41.7 × [actual bodyweight (ABW)/ideal bodyweight
(IBW)] Ideal body weight was calculated using the Lorentz formula
(12): height (cm) –100– {[height (cm) –150]/4} for males or
height (cm) –100– ([height (cm) –150]/2.5) for females. If the
ratio of actual body weight (kg) to ideal body weight (kg)
was ≥1, the assigned value was 1, as previously published (14). Frontiers in Nutrition Patient characteristics Between January 2010 and December 2020, our study
enrolled 595 individuals. The basic information was detailed
in Table 1. The median age was 70 (interquartile range 63–
77) years and 63% of the population were men. The median
PNI and GNRI scores at baseline were 46.9 (interquartile range
43.0–50.3), 102.1 (interquartile range 96.2–108.5), respectively. The BMI was 23.7 (interquartile range 21.9–26.2) kg/m2. The hypertension, DM, ICM accounted for 57, 23, and 26%,
respectively. In our study, more patients were in high risk of
malnutritional status either measured by PNI or GNRI (69 and
75%, respectively). The follow-up of adverse events continued until December
2021. The MCE was readmission to the hospital due to
worsening HF, myocardial infarction, stroke, or death from
any cause. Only the first occurrence was included in the
analysis for patients who experienced two or more events. The diagnosis of worsening HF was an unplanned hospital
admission or an urgent HF visit resulting in intravenous
therapy for HF. Myocardial infarction is used when the
evidence of myocardial injury (defined as an elevation of
cardiac troponin values with at least one value above the Frontiers in Nutrition 03 frontiersin.org Wang et al. 10.3389/fnut.2022.986731 TABLE 1 Characteristics of the study population. TABLE 1 Characteristics of the study population. TABLE 1 Characteristics of the study population. Patient characteristics Variable
Total
1PNI
1GNRI
Decreasing
nutritional
status
Increasing
nutritional
status
P
Decreasing
nutritional
status
Increasing
nutritional
status
P
Participants (n, %)
595 (100)
412 (69)
183 (31)
424 (71)
171 (29)
Men (n, %)
375 (63)
259 (63)
116 (63)
0.903
271 (64)
104 (62)
0.479
Age (y)
70 (63–77)
69 (63–76)
71 (63–77)
0.526
69 (62–76)
72 (64–78)
0.058
BMI (kg/m2)
23.7
(21.9–26.2)
23.7 (22.0–26.1)
23.5 (21.3–26.3)
0.286
23.7 (22.0–26.2)
23.7 (21.3–26.3)
0.273
Follow-up periods (years) 2.0 (1.0–4.0)
2.0 (1.0–4.0)
2.0 (1.0–3.8)
0.017
2.0 (1.0–3.9)
2.0 (1.0–4.0)
0.722
Lifestyle risk factors (n, %)
Smoking history
164 (28)
105 (25)
59 (32)
0.089
115 (27)
49 (29)
0.705
Drinking history
154 (26)
104 (25)
50 (27)
0.593
110 (26)
44 (26)
0.957
Concomitant disease (n, %)
Hypertension
342 (57)
242 (59)
100 (55)
0.351
247 (58)
95 (56)
0.547
DM
136 (23)
102 (25)
34 (19)
0.098
107 (25)
29 (17)
0.030
ICM
152 (26)
104 (25)
48 (26)
0.799
109 (26)
43 (25)
0.887
Echocardiography
LVEF (%)
61.4
(39.0–68.6)
61.5 (42.0–68.6)
60.9 (37.0–67.4)
0.226
62.2 (41.3–69.4)
60.6 (37.0–66.9)
0.056
LAD (mm)
47.0
(42.0–52.0)
47.0 (42.0–52.0)
46.0 (41.0–52.0)
0.627
47.0 (42.0–52.0)
46.0 (41.0–52.0)
0.888
LVEDD (mm)
54.0
(49.0–60.0)
54.0 (49.0–60.0)
55.0 (49.0–60.0)
0.408
54.0 (49.0–60.0)
55.0 (49.0–62.0)
0.188
MR
1.0 (1.0–2.0)
1.0 (0.6–2.0)
1.0 (1.0–2.0)
0.033
1.0 (1.0–2.0)
1.0 (1.0–2.0)
0.018
TR
1.0 (0.0–2.0)
1.0 (0.0–2.0)
1.0 (0.5–2.0)
0.998
1.0 (0.0–2.0)
1.0 (0.0–2.0)
0.896
Laboratory metrics
TC (mmol/L)
4.1 (3.4–4.8)
4.0 (3.4–4.7)
4.2 (3.5–5.0)
0.065
4.1 (3.4–4.8)
4.1 (3.4–5.0)
0.408
TG (mmol/L)
1.2 (0.9–1.7)
1.2 (0.9–1.6)
1.2 (0.9–1.8)
0.461
1.2 (0.9–1.6)
1.3 (0.9–1.8)
0.307
HDL-c (mmol/L)
1.0 (0.9–1.2)
1.0 (0.8–1.1)
1.1 (0.9–1.3)
0.000
1.0 (0.8–1.2)
1.1 (0.9–1.3)
0.001
LDL-c (mmol/L)
2.2 (1.7–2.8)
2.2 (1.7–2.8)
2.3 (1.7–2.9)
0.611
2.2 (1.7–2.8)
2.3 (1.7–2.9)
0.899
AST (U/L)
24 (20–31)
23 (20–31)
25 (21–31)
0.158
24 (20–31)
24 (20–31)
0.581
WBC (109/L)
6.1 (5.0–7.4)
6.1 (5.0–7.4)
6.1 (5.1–7.4)
0.849
6.1 (5.1–7.5)
6.0 (4.8–7.2)
0.268
PLT (109/L)
186
(153–225)
185 (153–227)
188 (150–223)
0.830
185 (153–227)
188 (148–223)
0.602
Hb (g/L)
130
(119–142)
129 (119–140)
130 (120–142)
0.238
130 (120–142)
129 (118–141)
0.620
Types of the pacemakers
Single-chamber
13 (2.2)
7 (3.8)
6 (1.5)
0.129
7 (4.1)
6 (1.4)
0.087
Dual-chamber
334 (56.1)
97 (53.0)
237 (57.5)
0.350
85 (49.7)
249 (58.7)
0.055
CRT-P
97 (16.3)
28 (15.3)
69 (16.7)
0.748
33 (19.3)
64 (15.1)
0.257
CRT-D
133 (22.4)
45 (24.6)
88 (21.4)
0.443
40 (23.4)
93 (21.9)
0.781
Dual-ICD
18 (3.0)
6 (3.3)
12 (2.9)
0.998
93 (21.9)
12 (2.8)
0.862
Pacing indications
AVB
144 (24.2)
36 (19.7)
108 (26.2)
0.106
35 (20.5)
109 (25.7)
0.213
SSS
125 (21.0)
34 (18.6)
91 (22.1)
0.390
28 (16.4)
97 (22.9)
0.099
AF with bradycardia
104 (17.5)
34 (18.6)
70 (17.0)
0.723
31 (18.1)
73 (17.2)
0.884
LBBB with HF
63 (15.3)
26 (14.2)
89 (15.0)
0.828
51 (29.8)
82 (19.3)
0.998
AVNA
80 (19.4)
53 (29.0)
133 (22.4)
0.013
26 (15.2)
63 (14.9)
0.008
Data were presented as percentages for categorical variables and mean ± standard deviation (SD) or median and interquartile range (IQR) for continuous variables. Frontiers in Nutrition Patient characteristics Group differences
were evaluated using Student t-tests or Mann–Whitney U tests for continuous variables and chi-square or Fisher exact tests for categorical variables. PNI, prognostic nutritional index; GNRI, geriatric nutritional risk index; BMI, body mass index; DM, diabetes mellitus; ICM, ischemic cardiomyopathy; LVEF, left ventricular ejection
fraction; LAD, left atrial diameter; LVEDD, left ventricular end-diastolic diameter; MR, mitral regurgitation; TR, tricuspid regurgitation; TC, total cholesterol; TG, triglyceride; HDL-c,
high-density lipoprotein- cholesterol; LDL-c, low-density lipoprotein- cholesterol; AST, aspartate transaminase; WBC, white blood cell; PLT, platelet; Hb, hemoglobin; CRT-P, cardiac
resynchronization therapy-pacemaker; CRT-D, cardiac resynchronization therapy- defibrillator; dual-ICD, dual-implantable cardioverter defibrillator; AVB, atrioventricular block; SSS,
sick sinus syndrome; AF, atrial fibrillation; LBBB, left bundle branch block; HF, heart failure; AVNA, atrial fibrillation with atrioventricular node ablation. 1PNI 04 Frontiers in Nutrition frontiersin.org Wang et al. 10.3389/fnut.2022.986731 FIGURE 1
Kaplan–Meier survival analysis for major adverse cardiovascular event (MCE) stratified by 1PNI (A) and 1GNRI (B). PNI, prognostic nutritional
index; GNRI, geriatric nutritional risk index. FIGURE 1
Kaplan–Meier survival analysis for major adverse cardiovascular event (MCE) stratified by 1PNI (A) and 1GNRI (B). PNI, prognostic nutritional
index; GNRI, geriatric nutritional risk index. TABLE 2 The association of 1PNI, 1GNRI with major adverse cardiovascular event (MCE) risk. TABLE 2 The association of 1PNI, 1GNRI with major adverse cardiovascular event (MCE) risk. Association between the 1PNI and MCE risk
Association between the 1GNRI and MCE risk
HR
95%CI
P
HR
95%CI
P
Model 1
2.056
1.372–3.082
0.000
1.943
1.295–2.914
0.001
Model 2
2.045
1.364–3.065
0.001
1.913
1.271–2.879
0.002
Model 3
2.055
1.370–3.083
0.000
1.915
1.272–2.884
0.002
Model 4
2.228
1.482–3.350
0.000
2.178
1.439–3.295
0.000
We performed Cox Proportional Hazards models of patients stratified according to PNI and GNRI improved or decreased, respectively. Model 1: Unadjusted. Model 2: Adjusted for gender, age range, and BMI (overweight/obesity). Model 3: Adjusted for gender, age range, BMI, and life risk factors (smoking, current drinking). Model 4: Adjusted for gender, age range, life risk factors, and baseline health status (diabetes, hypertension, ischemic cardiomyopathy and hyperlipidemia). Association between the 1PNI and MCE risk Model 3: Adjusted for gender, age range, BMI, and life risk factors (smoking, current drinking). Model 4: Adjusted for gender, age range, life risk factors, and baseline health status (diabetes, hypertension, ischemic cardiomyopathy and hyperlipidemia). 3%). The most common reasons for pacing were AVB,
AF with AVNA, and SSS, accounting 24, 19, and 21%,
respectively. Patient characteristics AF with slow ventricular rate was 17% and LBBB
with HF was 18%. The
types
of
the
pacemakers
were
single-chamber
(2%),
dual-chamber
(56%),
cardiac-resynchronization
therapy-pacemaker (CRT-P, 16%), CRT-defibrillator (22%),
dual-implantable
cardioverter
defibrillator
(dual-ICD, 05 frontiersin.org frontiersin.org Wang et al. 10.3389/fnut.2022.986731 Nutritional status and major adverse
cardiovascular event risk without overall overweight/obesity, patients with or without
hypertension or diabetes or ischemic cardiomyopathy or
hyperlipidemia (Figures 2, 3). ROC curve analysis revealed an AUC of 0.586 (sensitivity:
45.3%, specificity: 72%) for 1PNI scores and AUC of
0.592 (sensitivity: 44.2%, specificity: 74.2%) for 1GNRI
scores, but their predictive abilities were not statistically
different (Figures 3, 4). All clinical follow-up data were obtained. The median
follow-up period was 2.0 (1.0–4.0) years. During the follow-
up, 95 (16%) MCEs were identified. Figure 1 shows Kaplan–
Meier curves for MCE among patients stratified by 1PNI and
1GNRI scores. In high risk of malnutritional status groups, the
cumulative incidence of MCEs clearly increased (log-rank test,
P < 0.0001 each). Discussion Table 2 shows Cox proportional hazard analyses for MCEs. On unadjusted Cox modeling, rates of MCEs rose progressively
with low risk of malnutrition groups, even after adjusting for
other risk factors (PNI scores < 0: HR: 2.228, 95% CI 1.482–
3.350, P < 0.000; GNRI scores < 0: HR: 2.178, 95% CI 1.439–
3.295, P < 0.000). The purpose of this study was to determine whether the
changes in nutritional status, as measured by 1PNI and 1GNRI
scores, was associated with MCEs in patients with PMI. The
key finding of the present study was that a decrease in PNI or
GNRI score during the follow-up was significantly and strongly
associated with a higher risk of MCE. The changes in PNI
and GNRI scores had similar predictive power for MCEs in
our study population. To our knowledge, this is the first study Decreased PNI or GNRI scores were consistently associated
with worse long-term clinical outcomes in each subgroup,
including
male
or
female,
age
≥
65
or
<
65
years,
with or without smoking and drinking history, with or FIGURE 2
Interaction of confounding factors on the association between 1PNI and major adverse cardiovascular event (MCE) risk. PNI, prognostic
nutritional index. FIGURE 2
Interaction of confounding factors on the association between 1PNI and major adverse cardiovascular event (MCE) risk. PNI, prognostic
nutritional index. 06 Frontiers in Nutrition frontiersin.org Wang et al. 10.3389/fnut.2022.986731 FIGURE 3
Interaction of confounding factors on the association between 1GNRI and major adverse cardiovascular event (MCE) risk. GNRI, geriatric
nutritional risk index. FIGURE 3
Interaction of confounding factors on the association between 1GNRI and major adverse cardiovascular event (MCE) risk. GNRI, geriatric
nutritional risk index. FIGURE 3
Interaction of confounding factors on the association between 1GNRI and major adverse cardiovascular event (MCE) risk. GNRI, geriatric
nutritional risk index. FIGURE 3
Interaction of confounding factors on the association between 1GNRI and major adverse cardiovascular event (MCE) risk. GNRI, geriatric
nutritional risk index. to demonstrate the long-term predictive significance of the
changes in the nutritional status evaluated by PNI and GNRI
scores in patients with PMI. variety of conditions (20–22). The PNI score was suitable for
evaluating distinct characteristics of two components and was
straightforward to compute using affordable objective markers. Frontiers in Nutrition Discussion The GNRI score was also an objective nutritional evaluation
tool, calculated by a blood biomarker, height and weight, and
several recent studies have demonstrated a link between this
nutritional marker and cardiovascular risk in HF patients (12,
23). Considering the nutritional status was not still all the time,
we chose the changes of the PNI and GNRI scores to evaluating
their associations with MCEs. Previous
research
had
established
that
traditional
cardiovascular risk variables such as age, renal dysfunction,
male gender, a family history of coronary artery disease,
heart failure, and diabetes are predictors of death following
Pacemaker implantation (16). Additionally, more research had
been conducted on the association between nutritional status
and clinical outcomes (17, 18). In this study, we examined this
association using the changes in PNI and GNRI scores during
the follow-up. GNRI and PNI scores include the objective
quantitative data of the patient. In the calculation of the GNRI,
only the height, weight, and serum albumin data of the patient
are required. Similarly, the calculation of the PNI needs only the
serum albumin level and the lymphocyte count of the patient. A study enrolled patients undergoing cardiovascular surgery
and found that a low PNI score was substantially linked with
postoperative complications and survival and PNI score might
be a valuable and reliable tool for assessing nutritional status
before surgery, and it should be taken into account when
determining the indication for and strategy for cardiovascular
surgery (24). One study studied the association between PNI
score and stable coronary artery disease and discovered that
the PNI score was independently associated with higher
MCE rates (25). Another research analyzed the link between One study on gastrointestinal surgery (19) first described the
as an integrated and multiparameter nutritional management
paradigm in 1980. Since then, numerous studies had identified
as an independent risk factor for poor clinic outcomes in a 07 frontiersin.org Wang et al. 10.3389/fnut.2022.986731 FIGURE 4
Receiver operating characteristic (ROC) curve result for 1PNI and 1GNRI. Receiver operating characteristic (ROC) curve result for 1PNI and 1GNRI. acute HF and PNI score and found that PNI score was
independently associated with cardiovascular death and total
mortality (26). Yamada et al. (13) included 119 patients with
a CRT-device and divided the patients based on whether
their PNI score had increased at 6-month follow-up or
not after the procedure. Discussion The PNI scores raised group had
a lower occurrence of adverse events than the PNI scores
decreased group in the Kaplan-Meier analysis. Newly published
analysis (21) enrolling 141 patients with acute HF and
performing multivariate analysis which displayed that PNI
score in the improved group was independently associated
with good outcomes. These outcomes were consistent with
our studies. In addition to its use in the cardiovascular
field, PNI score may be an effective index for predicting the
rate of adverse events in other domains. A study enrolling
patients with small hepatocellular carcinoma who had liver
resection found that PNI score was an independent predictor of
overall survival and recurrence-free survival in a multivariable
study (27). predictability for mortality (30). Another analysis studied the
link of the changes in GNRI score and major adverse cardiac and
cerebrovascular events in incident peritoneal dialysis patients,
finding that patients with worsening or stationary GNRI score
were at significantly greater risk for adverse events. These
outcomes were well consistent with our results. In this study, the area under the curve (AUC) of 1PNI
and 1GNRI for determining MACs in patients with PMI was
0.586 and 0.592, respectively. However, these values may differ
based on the patient population. Thus, the prognostic efficacy
of 1PNI and 1GNRI in individuals with various cardiovascular
disorders may not be constant. In our study, we found the rates of MCE increased in both
unadjusted and adjusted Cox modeling in decreased PNI and
GNRI scores. Multivariate Cox hazard models demonstrated
that both 1PNI and 1GNRI scores was independently
associated with MCE after adjusting for risk factors. But we
found a significant association between either 1PNI or 1GNRI
score and age, which might be explained that the younger
people have faster metabolisms and higher nutritional needs
than the elderly. Once malnutrition occurred, it might be hit
harder in this group. Geriatric nutritional risk index (GNRI) score was a validated
nutritional assessment method screening nutritional conditions. A study investigated the association between GNRI score
and stable coronary artery disease and reported that GNRI
score ≤98 showed an increase in the incidences of cardiac
death or non-fatal MI compared with GNRI score > 98 (28). Minamisawa et al. (29) found patients with low GNRI score had
worse prognoses than those with high GNRI score in patients
at risk for HF. Frontiers in Nutrition Publisher’s note All claims expressed in this article are solely those of the
authors and do not necessarily represent those of their affiliated
organizations, or those of the publisher, the editors and the
reviewers. Any product that may be evaluated in this article, or
claim that may be made by its manufacturer, is not guaranteed
or endorsed by the publisher. Acknowledgments We would like to give our appreciation and thanks to
Yang B. (Institute of Lipids Medicine, Wenzhou Medical
University, Wenzhou, China; School of Public Health and
Management, Wenzhou Medical University, Wenzhou, China,
e-mail address: yb@wmu.edu.cn) for his constant help of
polishing and statistical guidance. Data availability statement The raw data supporting the conclusions of this article will
be made available by the authors, without undue reservation. Conflict of interest The authors declare that the research was conducted in the
absence of any commercial or financial relationships that could
be construed as a potential conflict of interest. Limitation Our
study
first
evaluated
the
long-term
predictive
significance of 1PNI and 1GNRI score in patients with
PMI. We also found that 1PNI and 1GNRI scores had similar
predictive power in our study. However, our study had several
limitations. First, as a single-center, observational study of a
small patient cohort, unknown confounding factors might have
affected the outcomes, regardless of analytical adjustments. Second, our population was limited to pacemaker implantation
and could not be generalized to other populations. Third,
considering the small population of the study, we did not
perform a subgroup analysis between PNI or GNRI scores and
HFH, MI, stroke, or death. Conclusion Improved malnutrition risk assessed by the either 1PNI or
1GNRI were associated with increased cardiovascular events in
PMI patients. The changes in PNI and GNRI scores had similar
predictive power and might be useful for risk stratification of
patients with pacing indications in clinical practice during the
follow-up. By evaluating these two tools, we can forecast the
probability of future adverse occurrences in patients during
clinical work, which might help us better treat patients. Author contributions CC and XH designed the study, investigated, and revised
the manuscript. KW and LL contributed to the investigation
and drafted the manuscript. CPC and MW contributed to
the investigation and prepared the figures. XH provided study
supervision. All authors contributed to critical revision of the
manuscript and approved its final version. Funding This
work
was
supported
by
the
National
Natural
Science Foundation of China (81900737) and the Basic
Scientific Research Program of Wenzhou Medical University,
China (KYYW202015). Discussion A study on hemodialysis concluded the 1GNRI
score could predict all-cause, cardiovascular mortality and prove Serum albumin, the most abundant plasma protein, was
synthesized in the liver and secreted into the vascular
space, where it was distributed throughout the body. It had
traditionally been used to assess nutritional status and visceral
protein synthesis function. Low levels of serum albumin had also
been determined to be an independent risk factor for survival
in patients after surgery (31, 32). Lymphocytes could be used Frontiers in Nutrition 08 frontiersin.org Wang et al. 10.3389/fnut.2022.986731 Affiliated Hospital of Wenzhou Medical University (YS2022-
306). The patients/participants provided their written informed
consent to participate in this study. to assess immunological nutritional status. Low lymphocytes
meant poor immune status. Weight loss related with the
development of cardiac cachexia was frequently connected with
decreases in physical function and a worse prognosis. Thus,
PNI score, combined serum albumin and lymphocyte could be
a good index for screening nutritional status. The GNRI score,
which was derived using serum albumin, ABW, and IBW while
controlling for body edema, was an accurate index for assessing
nutritional status. References 16. Rohacek M, Baddour LM. Cardiovascular implantable electronic device
infections: associated risk factors and prevention. Swiss Med Wkly. (2015)
145:w14157. 1. Glikson M, Nielsen JC, Kronborg MB, Tolosana JM. Corrigendum to: 2021
esc guidelines on cardiac pacing and cardiac resynchronization therapy: developed
by the task force on cardiac pacing and cardiac resynchronization therapy of
the European society of cardiology (ESC): with the special contribution of the
European heart rhythm association (EHRA). Europace. (2022) 24:699. doi: 10.1093/
europace/euac023 17. Balli M, Cetin M, Koksal F, Sag FE, Katkat F, Tekin EE, et al. Predictors
of pacemaker-induced cardiomyopathy and importance of nutritional status and
prognostic nutritional index. Acta Cardiol Sin. (2022) 38:151–8. doi: 10.6515/ACS. 202203_38(2).20211117A 2. Doshi RN, Daoud EG, Fellows C, Turk K, Duran A, Hamdan MH, et al. Left
ventricular-based cardiac stimulation post AV nodal ablation evaluation (the PAVE
study). J Cardiovasc Electrophysiol. (2005) 16:1160–5. doi: 10.1111/j.1540-8167. 2005.50062.x 18. Hayiroglu MI, Cinar T, Cinier G, Yüksel G, Pay L, Keskin K, et al. Prognostic
value of serum albumin for long-term mortality in patients with dual-chamber
permanent pacemakers. Biomark Med. (2022) 16:341–8. doi: 10.2217/bmm-2021-
0991 3. Varma N, Auricchio A, Connolly AT, Boehmer J, Bahu M, Costanzo MR,
et al. The cost of non-response to cardiac resynchronization therapy: characterizing
heart failure events following cardiac resynchronization therapy. Europace. (2021)
23:1586–95. doi: 10.1093/europace/euab123 19. Buzby GP, Mullen JL, Matthews DC, Hobbs CL, Rosato EF. Prognostic
nutritional index in gastrointestinal surgery. Am J Surg. (1980) 139:160–7. 20. Candeloro M, Di Nisio M, Balducci M, Genova S, Valeriani E, Pierdomenico
SD, et al. Prognostic nutritional index in elderly patients hospitalized for acute heart
failure. ESC Heart Fail. (2020) 7:2479–84. 4. Corish CA, Bardon LA. Malnutrition in older adults: screening and
determinants. Proc Nutr Soc. (2019) 78:372–9. 5. Norman K, Pichard C, Lochs H, Pirlich M. Prognostic impact of disease-
related malnutrition. Clin Nutr. (2008) 27:5–15. 21. Kawata T, Ikeda A, Masuda H, Komatsu S. Changes in prognostic nutritional
index during hospitalization and outcomes in patients with acute heart failure. Heart Vessels. (2022) 37:61–8. 6. Camanho LEM, Saad EB, Slater C, Oliveira Inacio Junior LA, Vignoli G,
Carvalho Dias L, et al. Clinical outcomes and mortality in old and very old patients
undergoing cardiac resynchronization therapy. PLoS One. (2019) 14:e0225612. doi:
10.1371/journal.pone.0225612 22. Dai Y, Liu M, Lei L, Lu S. Prognostic significance of preoperative prognostic
nutritional index in ovarian cancer: a systematic review and meta-analysis. References Medicine (Baltimore). (2020) 99:e21840. 7. Nagoshi
T,
Yoshimura
M. Nutritional
status
assessment
as
a
potential
prognostic
indicator
in
patients
before
the
development
of
symptomatic heart failure. Circ J. (2018) 82:1501–2. doi: 10.1253/circj.CJ-18-
0454 23. Hao XY, Li DY, Zhang N. Geriatric nutritional risk index as a predictor for
mortality: a meta-analysis of observational studies. Nutr Res. (2019) 71:8–20. 24. Hayashi J, Uchida T, Ri S, Hamasaki A, Kuroda Y, Yamashita A, et al. Clinical significance of the prognostic nutritional index in patients undergoing
cardiovascular surgery. Gen Thorac Cardiovasc Surg. (2020) 68:774–9. 8. Dent E, Hoogendijk EO, Visvanathan R, Wright ORL. Malnutrition screening
and assessment in hospitalised older people: a review. J Nutr Health Aging. (2019)
23:431–41. 25. Wada H, Dohi T, Miyauchi K, Jun S, Endo H, Doi S, et al. Relationship
between the prognostic nutritional index and long-term clinical outcomes in
patients with stable coronary artery disease. J Cardiol. (2018) 72:155–61. doi: 10. 1016/j.jjcc.2018.01.012 9. Sharma Y, Avina P, Ross E, Horwood C, Hakendorf P, Thompson C. Validity
of the malnutrition universal screening tool for evaluation of frailty status
in older hospitalised patients. Gerontol Geriatr Med. (2022) 8:2333721422110
7817. 26. Cheng YL, Sung SH, Cheng HM, Hsu PF, Guo CY, Yu WC, et al. Prognostic
nutritional index and the risk of mortality in patients with acute heart failure. J Am
Heart Assoc. (2017) 6:e004876. 10. Abd Aziz NAS, Teng NIMF, Hamid MRA, Ismail NH. Assessing
the
nutritional
status
of
hospitalized
elderly. Clin
Interv
Aging. (2017)
12:1615–25. 27. Peng W, Li C, Wen TF, Yan LN, Li B, Wang WT, et al. Postoperative
prognostic nutritional index change is an independent predictor of survival in
patients with small hepatocellular carcinoma. Am J Surg. (2016) 212:122–7. doi:
10.1016/j.amjsurg.2015.06.023 11. Li HD, Cen KD, Sun WF, Feng B. Prognostic value of geriatric nutritional
risk index in elderly patients with heart failure: a meta-analysis. Aging Clin Exp Res. (2021) 33:1477–86. 28. Kunimura A, Ishii H, Uetani T, Aoki T, Harada K, Hirayama K, et al. Impact
of geriatric nutritional risk index on cardiovascular outcomes in patients with stable
coronary artery disease. J Cardiol. (2017) 69:383–8. 12. Bouillanne O, Morineau G, Dupont C, Coulombel I, Vincent JP, Nicolis
I, et al. Geriatric nutritional risk Index: a new index for evaluating at-risk
elderly medical patients. Am J Clin Nutr. (2005) 82:777–83. doi: 10.1093/ajcn/82. 4.777 29. Ethics statement The studies involving human participants were reviewed
and approved by the Institutional Review Board of the First Frontiers in Nutrition 09 frontiersin.org 10.3389/fnut.2022.986731 Wang et al. 10.3389/fnut.2022.986731 References Minamisawa M, Miura T, Motoki H, Ueki Y, Mochidome T, Oguchi Y, et al. Geriatric nutritional risk index predict cardiovascular events in patients at risk for
heart failure. Circulation. (2016) 134:A13099. 13. Yamada S, Yoshihisa A, Hijioka N, Amami K, Kaneshiro T, Ishida T,
et al. Associations of the prognostic nutritional index with the cardiac function
and survival after cardiac resynchronization therapy. Internal Med. (2021)
60:985–91. 30. Yajima T, Yajima K, Takahashi H. Impact of annual change in geriatric
nutritional risk index on mortality in patients undergoing hemodialysis. Nutrients. (2020) 12:3333. doi: 10.3390/nu12113333 14. González Ferreiro R, López Otero D, Álvarez Rodríguez L, Otero García Ó,
Pérez Poza M, Antúnez Muiños PJ, et al. Prognostic impact of change in nutritional
risk on mortality and heart failure after transcatheter aortic valve replacement. Circ
Cardiovasc Interv. (2021) 14:e009342. doi: 10.1161/CIRCINTERVENTIONS.120. 009342 14. González Ferreiro R, López Otero D, Álvarez Rodríguez L, Otero García Ó,
Pérez Poza M, Antúnez Muiños PJ, et al. Prognostic impact of change in nutritional
risk on mortality and heart failure after transcatheter aortic valve replacement. Circ
Cardiovasc Interv. (2021) 14:e009342. doi: 10.1161/CIRCINTERVENTIONS.120. 009342 31. Yin M, Si L, Qin WD, Li C, Zhang J, Yang H, et al. Predictive value of serum
albumin level for the prognosis of severe sepsis without exogenous human albumin
administration: a prospective cohort study. J Intensive Care Med. (2018) 33:687–94. doi: 10.1177/0885066616685300 15. Collet JP, Thiele H, Barbato E, Barthélémy O, Bauersachs J, Bhatt DL,
et al. 2020 ESC guidelines for the management of acute coronary syndromes in
patients presenting without persistent ST-segment elevation. Eur Heart J. (2021)
42:1289–367. 32. Huang W, Li CZ, Wang ZQ, Wang H, Zhou N, Jiang J, et al. Decreased
serum albumin level indicates poor prognosis of COVID-19 patients: hepatic injury
analysis from 2,623 hospitalized cases. Sci China Life Sci. (2020) 63:1678–87. doi:
10.1007/s11427-020-1733-4 Frontiers in Nutrition frontiersin.org 10
|
https://openalex.org/W4205559996
|
https://www.frontiersin.org/articles/10.3389/fcell.2021.816335/pdf
|
English
| null |
A Novel Potent Carrier for Unconventional Protein Export in Ustilago maydis
|
Frontiers in cell and developmental biology
| 2,022
|
cc-by
| 10,592
|
A Novel Potent Carrier for
Unconventional Protein Export in
Ustilago maydis
Magnus Philipp, Kai P. Hussnaetter, Michèle Reindl, Kira Müntjes, Michael Feldbrügge and
Kerstin Schipper*
Institute for Microbiology, Heinrich Heine University Düsseldorf, Düsseldorf, Germany Institute for Microbiology, Heinrich Heine University Düsseldorf, Düsseldorf, Germany Recombinant proteins are ubiquitously applied in fields like research, pharma, diagnostics
or the chemical industry. To provide the full range of useful proteins, novel expression hosts
need to be established for proteins that are not sufficiently produced by the standard
platform organisms. Unconventional secretion in the fungal model Ustilago maydis is an
attractive novel option for export of heterologous proteins without N-glycosylation using
chitinase Cts1 as a carrier. Recently, a novel factor essential for unconventional Cts1
secretion termed Jps1 was identified. Here, we show that Jps1 is unconventionally
secreted using a fusion to bacterial β-glucuronidase as an established reporter. Interestingly, the experiment also demonstrates that the protein functions as an
alternative carrier for heterologous proteins, showing about 2-fold higher reporter
activity than the Cts1 fusion in the supernatant. In addition, Jps1-mediated secretion
even allowed for efficient export of functional firefly luciferase as a novel secretion target
which could not be achieved with Cts1. As an application for a relevant pharmaceutical
target, export of functional bi-specific synthetic nanobodies directed against the SARS-
CoV2 spike protein was demonstrated. The establishment of an alternative efficient carrier
thus constitutes an excellent expansion of the existing secretion platform. ORIGINAL RESEARCH
published: 10 January 2022
doi: 10.3389/fcell.2021.816335 Edited by: Edited by:
Julien Villeneuve,
Institute for Functional Genomics
(IGF), France Reviewed by:
Carmen Faso,
University of Bern, Switzerland
Kai Heimel,
University of Göttingen, Germany
Rosana Puccia,
Federal University of São Paulo, Brazil Reviewed by:
Carmen Faso,
University of Bern, Switzerland
Kai Heimel,
University of Göttingen, Germany
Rosana Puccia,
Federal University of São Paulo, Brazil Reviewed by:
Carmen Faso,
University of Bern, Switzerland
Kai Heimel,
University of Göttingen, Germany
Rosana Puccia,
Federal University of São Paulo, Brazil
*Correspondence:
Kerstin Schipper
kerstin.schipper@uni-
duesseldorf.de Reviewed by:
Carmen Faso,
University of Bern, Switzerland
Kai Heimel,
University of Göttingen, Germany
Rosana Puccia,
Federal University of São Paulo, Brazil
*Correspondence:
Kerstin Schipper
kerstin.schipper@uni- Keywords: luciferase, anti-Sars-CoV2 nanobody, unconventional secretion, Ustilago maydis, sybody INTRODUCTION Specialty section:
This article was submitted to
Membrane Traffic,
a section of the journal
Frontiers in Cell and Developmental
Biology
Received: 16 November 2021
Accepted: 17 December 2021
Published: 10 January 2022 Specialty section:
This article was submitted to
Membrane Traffic,
a section of the journal
Frontiers in Cell and Developmental
Biology The market for recombinant proteins like biopharmaceuticals is steadily increasing (Walsh 2018). As
one example, the number of monoclonal antibody therapeutics entering phase 3 clinical trials has
risen from 39 in 2014 to 88 in 2020 (Reichert 2015; Kaplon and Reichert 2021). Protein secretion into
the culture broth is an excellent strategy for the production of recombinant proteins because it
supports straight-forward and inexpensive downstream processing (Nicaud et al., 1986; Flaschel and
Friehs 1993). In eukaryotes, proteins are mostly targeted via the endomembrane system by
N-terminal signal peptides for secretion (Viotti 2016). By contrast, the term unconventional
secretion describes protein export that does not occur via the classical endomembrane system
including endoplasmic reticulum and Golgi apparatus (Nickel 2010). Various routes for such
alternative secretion events exist, including direct transfer across the plasma membrane via
transporters or self-sustained translocation or vesicular pathways where membrane vesicles are
hitchhiked for export (Nickel 2010; Rabouille 2017). Received: 16 November 2021 Accepted: 17 December 2021 Published: 10 January 2022 Citation: Philipp M, Hussnaetter KP, Reindl M,
Müntjes K, Feldbrügge M and
Schipper K (2022) A Novel Potent
Carrier for Unconventional Protein
Export in Ustilago maydis. Front. Cell Dev. Biol. 9:816335. doi: 10.3389/fcell.2021.816335 Philipp M, Hussnaetter KP, Reindl M,
Müntjes K, Feldbrügge M and
Schipper K (2022) A Novel Potent
Carrier for Unconventional Protein
Export in Ustilago maydis. Front. Cell Dev. Biol. 9:816335. doi: 10.3389/fcell.2021.816335 Philipp M, Hussnaetter KP, Reindl M,
Müntjes K, Feldbrügge M and
Schipper K (2022) A Novel Potent
Carrier for Unconventional Protein
Export in Ustilago maydis. Front. Cell Dev. Biol. 9:816335. doi: 10.3389/fcell.2021.816335 Unconventional export of chitinase Cts1 in yeast cells of the fungal model Ustilago maydis is
coupled to cytokinesis in a lock-type mechanism (Reindl et al., 2019). Upon formation of the January 2022 | Volume 9 | Article 816335 Frontiers in Cell and Developmental Biology | www.frontiersin.org 1 Philipp et al. New Carrier for Protein Export FIGURE 1 | Jps1 is unconventionally secreted and serves as an alternative carrier for Gus export. (A) Schematic display of the proteins expressed to study
unconventional secretion. Cytoplasmic Gus (Guscyt) is used as a lysis control (top). Gus-Jps1 (middle) and Gus-Cts1 (bottom) harbor the respective carrier proteins at the
C-terminus. All proteins carry an SHH (double Strep, ten times His, triple HA) tag indicated in black (Sarkari et al., 2014). All schemes are drawn to scale. (B) Enzymatic
reaction mediated by β-glucuronidase. 4-methyl-umbeliferyl-β-D-glucuronide (4-MUG) and H2O are converted to 4-methyl-umbelliferone which is a fluorescent
molecule (365 nm excitation/465 nm emission). (C) Determination of intracellular Gus activity. Progenitor strain AB33P8Δ (Ctrl) and AB33 Guscyt expressing cytoplasmic
Gus were included as controls. The experiment was conducted in three biological replicates. (D) Comparative extracellular Gus activity of strains using either Cts1 or
Jps1 as a carrier. Enzyme activities were normalized to average values of the strain secreting Gus-Cts1. AB33P8Δ and AB33 Guscyt were used as a negative and lysis
controls, respectively. The experiment was conducted in three biological replicates. (E) Representative Western blot analysis of Gus-Cts1 and Gus-Jps1 secretion. Extracellular protein was enriched from culture supernatants by TCA precipitation. Intracellular protein levels were visualized by cell extracts. Western blots show 1 ml of
precipitated supernatants (TCA) and 10 μg cell extract (CE). Full length protein signal indicated by arrows, degradation bands with a rhombus. (F) Quantification of
secreted protein using Western blot analysis. Citation: Supernatants of strains producing Gus-Jps1 or Gus-Cts1 were enriched by TCA precipitation and subjected to Western
blot analysis. Signal intensities were compared to defined protein amounts of Multiple Tag protein (GenScript Piscataway, NJ, United States) included in the same gel. Bars show extrapolated protein amounts in µg/L. Western blots used for the analysis, see Supplementary Figure S3. Three biological replicates are shown; error bars
in figures (C), (D), and (F) indicate standard deviation. Definition of statistical significance (***): p-value < 0.05. p-value derived from Student’s unpaired t-test. FIGURE 1 | Jps1 is unconventionally secreted and serves as an alternative carrier for Gus export. (A) Schematic display of the proteins expressed to study
unconventional secretion. Cytoplasmic Gus (Guscyt) is used as a lysis control (top). Gus-Jps1 (middle) and Gus-Cts1 (bottom) harbor the respective carrier proteins at the
C-terminus. All proteins carry an SHH (double Strep, ten times His, triple HA) tag indicated in black (Sarkari et al., 2014). All schemes are drawn to scale. (B) Enzymatic
reaction mediated by β-glucuronidase. 4-methyl-umbeliferyl-β-D-glucuronide (4-MUG) and H2O are converted to 4-methyl-umbelliferone which is a fluorescent
molecule (365 nm excitation/465 nm emission). (C) Determination of intracellular Gus activity. Progenitor strain AB33P8Δ (Ctrl) and AB33 Guscyt expressing cytoplasmic
Gus were included as controls. The experiment was conducted in three biological replicates. (D) Comparative extracellular Gus activity of strains using either Cts1 or
Jps1 as a carrier. Enzyme activities were normalized to average values of the strain secreting Gus-Cts1. AB33P8Δ and AB33 Guscyt were used as a negative and lysis
controls, respectively. The experiment was conducted in three biological replicates. (E) Representative Western blot analysis of Gus-Cts1 and Gus-Jps1 secretion. Extracellular protein was enriched from culture supernatants by TCA precipitation. Intracellular protein levels were visualized by cell extracts. Western blots show 1 ml of
precipitated supernatants (TCA) and 10 μg cell extract (CE). Full length protein signal indicated by arrows, degradation bands with a rhombus. (F) Quantification of
secreted protein using Western blot analysis. Supernatants of strains producing Gus-Jps1 or Gus-Cts1 were enriched by TCA precipitation and subjected to Western
blot analysis. Signal intensities were compared to defined protein amounts of Multiple Tag protein (GenScript Piscataway, NJ, United States) included in the same gel. Bars show extrapolated protein amounts in µg/L. Western blots used for the analysis, see Supplementary Figure S3. Three biological replicates are shown; error bars
in figures (C), (D), and (F) indicate standard deviation. Citation: Definition of statistical significance (***): p-value < 0.05. p-value derived from Student’s unpaired t-test. because
it
avoids
post-translational
modifications
like
N-glycosylation occurring in the endomembrane system. In
addition, there is no apparent size limitation (Stock et al.,
2012). Successful
examples
are
secretion
of
functional
enzymes
like
β-glucuronidase
or
β-galactosidase,
and
antibody
formats
like
single-chain
variable
fragments
(scFv) or nanobodies (Stock et al., 2012; Sarkari et al.,
2014; Terfrüchte et al., 2017; Reindl et al., 2020). While the
secretion system is operational for several target proteins, low
yields in the µg per liter range are currently limiting its
applicability
(Terfrüchte
et
al.,
2017). Recently,
major
improvements were achieved by the generation of protease-
deficient production strains, usage of strong constitutive
promoters and medium optimization (Sarkari et al., 2014;
Terfrüchte et al., 2018). However, novel strategies to further
advance the system are needed. daughter cell at one growth pole of the cigar shaped mother cell,
Cts1 is targeted to the so-called fragmentation zone delimited at
the mother-daughter neck by consecutive formation of two septa
(Langner et al., 2015). Here, the chitinase participates in
separation of the two cells likely by degrading the remnant cell
wall (Langner et al., 2015). Two septation factors, guanine
nucleotide exchange factor (GEF) Don1 and kinase Don3, are
essential for formation of the secondary septum and for Cts1
secretion (Weinzierl et al., 2002; Aschenbroich et al., 2019). Furthermore, a recently identified potential anchoring factor,
Jps1, is crucial for chitinase localization and export (Reindl
et al., 2020). Importantly,
unconventional
Cts1
secretion
can
be
exploited for co-export of heterologous proteins (Stock
et al., 2012). Circumventing the classical secretion system
is advantageous for the production of distinct proteins, January 2022 | Volume 9 | Article 816335 Frontiers in Cell and Developmental Biology | www.frontiersin.org 2 New Carrier for Protein Export Philipp et al. FIGURE 2 | Inducible secretion of Gus-Jps1 via transcriptional
regulation of don3. (A) Schematic display of the inducible secretion system. don3-gfp is expressed under control of the arabinose-inducible promoter Pcrg. Under glucose conditions the promoter is in its “off state”,
unconventionally secreted proteins under control of Poma are thus expressed
but not secreted Under arabinose condition the promoter is in its “on state” In the present study we demonstrate that Jps1 is a novel potent
carrier for co-export of heterologous proteins. Jps1 is a Potent New Carrier for
Unconventional Protein Export Jps1 is a Potent New Carrier for
Unconventional Protein Export p
Previous experiments had shown that Jps1 co-localizes with Cts1
in the fragmentation zone (Reindl et al., 2020), suggesting that it
might also be unconventionally secreted. To study this, we
applied the well-established β-glucuronidase (Gus) reporter
system
(Figure
1A,B). This
bacterial
enzyme
is
largely
inactivated
upon
secretion
through
the
eukaryotic
endomembrane
system. By
contrast,
it
is
released
in
a
functional state via unconventional secretion in yeast cells of
U. maydis (Stock et al., 2012). To assay unconventional secretion
of Jps1, a strain expressing a Gus-Jps1 fusion protein was
generated in the background of the octuple protease-deletion
laboratory strain AB33P8Δ (Figure 1A) (Terfrüchte et al., 2018). Microscopic analysis revealed that yeast cells expressing Gus-Jps1
did not show any morphological differences as compared to the
progenitor (Supplementary Figures S1, S2). The Gus-Jps1 fusion
did also not disturb Cts1 function as detected by determining
extracellular chitinase activity of AB33P8Δ/Gus-Jps1 which was
similar to the activity detected in a strain expressing Gus-Cts1
(Supplementary
Figure
S1). Subsequently,
intra-
and
extracellular Gus activity was determined (Figures 1C,D). The
progenitor strain AB33P8Δ was used as a negative control, while a
strain expressing intracellular Gus served as a lysis control (AB33
Guscyt) (Stock et al., 2012). High Gus activity was present in cell
extracts of all strains harboring the Gus enzyme but not in the
progenitor
AB33P8Δ
lacking
the
enzyme
(Figure
1C). Importantly, Gus activity was also detected in the supernatant
of Gus-Jps1 expressing strains but not for the lysis control,
confirming unconventional secretion of Jps1 (Figure 1D). At
the same time, this experiment demonstrates, that Jps1—similar
to Cts1—is able to act as a carrier for heterologous proteins. Notably, extracellular Gus activity levels were increased by about
2-fold in culture supernatants of Gus-Jps1 compared to Gus-Cts1
expressing strains (Figure 1D), suggesting that Jps1 might
constitute a more effective carrier than Cts1. Both strains were
also compared in terms of growth speed and strain fitness using
online monitoring in a BioLector device (m2p-labs, Baesweiler,
Germany) (Funke et al., 2010). The progenitor strain AB33P8Δ as
well as AB33P8Δ/Gus-Cts1 and AB33P8Δ/Gus-Jps1 showed
similar proliferation patterns and doubling times of about 3 h FIGURE 2 | Inducible secretion of Gus-Jps1 via transcriptional
regulation of don3. (A) Schematic display of the inducible secretion system. don3-gfp is expressed under control of the arabinose-inducible promoter Pcrg. Citation: We observed
improved overall yields of secreted protein and export of
firefly luciferase that was not functionally secreted via Cts1-
fusions. As a proof-of-principle for pharmaceutical proteins we
exported functional nanobodies directed against the receptor-
binding domain (RBD) of the SARS-CoV2 spike protein. The
novel carrier thus constitutes an important improvement of
our expression system towards a competitive production
platform. RESULTS Jps1 is a Potent New Carrier for
Unconventional Protein Export Jps1 is a Potent New Carrier for
Unconventional Protein Export Under glucose conditions the promoter is in its “off state”,
unconventionally secreted proteins under control of Poma are thus expressed
but not secreted. Under arabinose condition the promoter is in its “on state”
and proteins are secreted. Gus is fused to either Cts1 or Jps1 including an
internal SHH tag (double Strep, ten times His, triple HA). (B) Gus activity in
culture supernatants of AB33 derivatives expressing Gus-Cts1 or Gus-Jps1
and their Δdon3 variants. Enzymatic activity was normalized to average values
of positive controls secreting Gus-Cts1 constitutively. The diagram represents
the results of three biological replicates. Error bars depict standard deviation. Fold change of induced cultures depicted over brackets. Definition of
statistical significance (***): p-value < 0.05. p-value derived from Student’s
unpaired t-test. FIGURE 2 | Inducible secretion of Gus-Jps1 via transcriptional
regulation of don3. (A) Schematic display of the inducible secretion system. don3-gfp is expressed under control of the arabinose-inducible promoter Pcrg. Under glucose conditions the promoter is in its “off state”, unconventionally secreted proteins under control of Poma are thus expressed
but not secreted. Under arabinose condition the promoter is in its “on state”
and proteins are secreted. Gus is fused to either Cts1 or Jps1 including an
internal SHH tag (double Strep, ten times His, triple HA). (B) Gus activity in
culture supernatants of AB33 derivatives expressing Gus-Cts1 or Gus-Jps1
and their Δdon3 variants. Enzymatic activity was normalized to average values
of positive controls secreting Gus-Cts1 constitutively. The diagram represents
the results of three biological replicates. Error bars depict standard deviation. Fold change of induced cultures depicted over brackets. Definition of
statistical significance (***): p-value < 0.05. p-value derived from Student’s
unpaired t-test January 2022 | Volume 9 | Article 816335 Frontiers in Cell and Developmental Biology | www.frontiersin.org 3 Philipp et al. New Carrier for Protein Export FIGURE 3 | Efficient Jps1-mediated export of firefly luciferase as a new reporter for unconventional secretion. (A) Schematic display of the proteins expressed to
study unconventional secretion. Cytoplasmic FLuc (FLuccyt) was used as a lysis control (top). FLuc-Jps1 (middle) and FLuc-Cts1 (bottom) harbor the respective carrier
proteins at the C-terminus. All proteins carry an SHH tag indicated in black (Sarkari et al., 2014). All schemes are drawn to scale. (B) Enzymatic reaction mediated by
firefly luciferase: D-Luciferin and ATP are converted to oxiluciferin, AMP and CO2. Jps1 is a Potent New Carrier for
Unconventional Protein Export Definition of statistical significance (***): p-value < 0.05. p-value was
derived from Student’s unpaired t-test. (E) Representative Western blot of FLuc-Cts1 and FLuc-Jps1. Secreted protein was enriched from the supernatant by TCA
precipitation. Intracellular protein levels were visualized by cell extracts. Western blots show 1 ml of precipitated supernatants (TCA) and 10 μg cell extracts (CE). Full
length protein signals indicated by arrows, degradation bands with a rhombus. about 103 μg/L (about 2.7-fold increase; Figure 1F). In
summary, these results demonstrate that Jps1 can deal as a
powerful carrier for heterologous proteins with elevated levels
in comparison to Cts1. during the exponential growth phase when incubated in CM
medium supplemented with 1% glucose (Supplementary Figure
S2). Thus, Jps1 constitutes a promising candidate for a novel
potent carrier for heterologous proteins. To assay secretion on the protein level, Western blot
analyses were conducted. These experiments showed that
extracellular amounts of Gus-Jps1 were markedly increased
as compared to Gus-Cts1, while intracellular levels were
comparable. This
confirms
that
Jps1
is
secreted
with
enhanced
efficiency
in
relation
to
Cts1
(Figure
1E,
Supplementary Figure S3). To quantify this result distinct
amounts
of
Multiple
Tag
protein
(GenScript
Biotech,
Piscataway,
NJ,
United
States)
were
included
(Supplementary Figure S4). Quantification of the Western
blot signals revealed that Gus-Cts1 levels in the supernatant
reach concentrations of 38 μg/L while Gus-Jps1 is present at Jps1 is a Potent New Carrier for
Unconventional Protein Export During this reaction excited intermediates emit energy in the form of light that can
be detected as bioluminescence. (C) Comparison of intracellular FLuc activity of the strains AB33P8Δ/FLuc-Cts1 and AB33P8Δ/FLuc-Jps1. Enzymatic activity was
normalized to average values of strain secreting FLuc-Cts1. The progenitor strain AB33P8Δ was used as a negative control. Strain AB33 FLuccyt with intracellular FLuc
expression dealt as positive control. Three biological replicates are shown. (D) Comparison of extracellular FLuc activity of strains harboring either Cts1 or Jps1 as a
carrier. Enzymatic activity was normalized to average values of strain secreting FLuc-Cts1. Strain AB33 FLuccyt with intracellular FLuc expression dealt as lysis control. Three biological replicates are shown. Error bars in figures (C) and (D) indicate standard deviation. Definition of statistical significance (***): p-value < 0.05. p-value was
derived from Student’s unpaired t-test. (E) Representative Western blot of FLuc-Cts1 and FLuc-Jps1. Secreted protein was enriched from the supernatant by TCA
precipitation. Intracellular protein levels were visualized by cell extracts. Western blots show 1 ml of precipitated supernatants (TCA) and 10 μg cell extracts (CE). Full
length protein signals indicated by arrows, degradation bands with a rhombus. FIGURE 3 | Efficient Jps1-mediated export of firefly luciferase as a new reporter for unconventional secretion. (A) Schematic display of the proteins expressed to
study unconventional secretion. Cytoplasmic FLuc (FLuccyt) was used as a lysis control (top). FLuc-Jps1 (middle) and FLuc-Cts1 (bottom) harbor the respective carrier
proteins at the C-terminus. All proteins carry an SHH tag indicated in black (Sarkari et al., 2014). All schemes are drawn to scale. (B) Enzymatic reaction mediated by
firefly luciferase: D-Luciferin and ATP are converted to oxiluciferin, AMP and CO2. During this reaction excited intermediates emit energy in the form of light that can
be detected as bioluminescence. (C) Comparison of intracellular FLuc activity of the strains AB33P8Δ/FLuc-Cts1 and AB33P8Δ/FLuc-Jps1. Enzymatic activity was
normalized to average values of strain secreting FLuc-Cts1. The progenitor strain AB33P8Δ was used as a negative control. Strain AB33 FLuccyt with intracellular FLuc
expression dealt as positive control. Three biological replicates are shown. (D) Comparison of extracellular FLuc activity of strains harboring either Cts1 or Jps1 as a
carrier. Enzymatic activity was normalized to average values of strain secreting FLuc-Cts1. Strain AB33 FLuccyt with intracellular FLuc expression dealt as lysis control. Three biological replicates are shown. Error bars in figures (C) and (D) indicate standard deviation. don3 Induced Secretion Further Enhances
Gus-Jps1 Secretion Recently, we have established a system that allows for the
induction of unconventional secretion via regulation of kinase
Don3, a gatekeeper of the fragmentation zone (Hussnaetter et al.,
2021). To this end we used a arabinose-inducible promoter to
control don3 expression, which is prerequisite for secondary
septum formation (Weinzierl et al., 2002). Unconventional
secretion is only functional with a functional fragmentation
zone consisting of two septa (Aschenbroich et al., 2019). Here January 2022 | Volume 9 | Article 816335 Frontiers in Cell and Developmental Biology | www.frontiersin.org 4 New Carrier for Protein Export Philipp et al. we reproduced these findings using Jps1 as a carrier as
demonstrated by a strain which carried genetic modifications
for transcriptional induction of don3 and expressed the Gus-Jps1
reporter as read-out (Figure 2A,B) (Hussnaetter et al., 2021). Although we observed a slightly higher background activity in
arabinose cultures, the induction was more than 18-fold and thus,
significantly higher than for using Cts1 as a carrier protein,
showing about 12-fold induction (Figure 2B). Furthermore,
Gus-activity was elevated 2.4-fold compared to induced Gus-
Cts1 secretion and more than 3-fold compared to regular Gus-
Cts1 secretion. Hence, inducible Jps1 constitutes a powerful tool
for unconventional secretion of heterologous proteins. Jps1
enables export of functional firefly luciferase. unclear. The size of the FLuc-Cts1 fusion protein is likely not
affecting its unconventional secretion, since larger fusions had been
successfully exported in earlier studies (Stock et al., 2012). Eventually, structural interferences or other unknown features of
this particular fusion lead to reduced protein production or its
instability. These results further emphasize the advantage of having
a second carrier for the unconventional secretion system at hands. Unconventional Secretion of Functional
Antibodies Against Sars-CoV2-Receptor
Binding Domain g
Next, we tested unconventional secretion of nanobodies directed
against the SARS-CoV2 spike protein receptor binding domain
(RBD) as a timely example of pharmaceutically relevant targets. Therefore, strains were generated in which two synergistic
synthetic nanobodies (sybodies) directed against the Sars-
CoV2 spike-RBD were combined (Walter et al., 2020). The bi-
specific sybody was tagged with a 10× His-linker for purification
and fused to either Cts1 or Jps1 for unconventional secretion
(AB33P8Δ/Sy68/15-Cts1 and AB33P8Δ/Sy68/15-Jps1) (Figure 4A). Western blot analyses confirmed that both fusion proteins were
synthesized. However, Sy68/15-Cts1 was produced at a lower level
compared to Sy68/15-Jps1. The latter showed stronger degradation
than observed for other Jps1 fusion proteins (see above). In
supernatants only a very faint signal was present for Sy68/15-
Cts1 while for Sy68/15-Jps1 a stronger signal and less degradation
than in cell extracts was detected (Figure 4B). Quantification
revealed an increase of about 18-fold in signal intensity for the
Jps1 full-length fusion compared to the Cts1 full-length fusion
(Supplementary Figure S3). Subsequently, the antigen-binding
activity of the sybody was determined via direct confrontation
with spike-RBD immobilized on ELISA plates and subsequent
detection with an antibody sandwich Figure 4C. Immobilized
bovine serum albumin (BSA) dealt as a negative control. ELISA
experiments using cell extracts demonstrated that both sybody-
fusion proteins were functional in detecting the cognate antigen. While the activity of Sy68/15-Cts1 was only slightly above baseline,
Sy68/15-Jps1 showed strong volumetric activity (Figure 4D). Next,
sybody-fusion proteins were IMAC purified from culture
supernatants
and
applied
to
ELISA
in
up
to
10-fold
concentrated solutions Figure 4E. While no activity could be
observed
for
Sy68/15-Cts1,
Sy68/15-Jps1
showed
volumetric
binding activity on the antigen, confirming the secretion of the
functional
sybody
fusion
protein. Thus,
pharmaceutically
relevant nanobodies were exported in their functional form
using Jps1 as a carrier for unconventional secretion. Jps1 Enables Export of Functional Firefly
Luciferase Photinus pyralis luciferase FLuc was recently established for
intracellular
use
in
U. maydis
(Müntjes
et
al.,
2020). Bioluminescence would be a straight-forward alternative read-
out for unconventional secretion because the signal can be
detected directly from the culture broth while the established
reporters Gus and β-galactosidase (LacZ) require more elaborate
biochemical assays. Further advantages are low background signals
and the use of the inexpensive substrate D-luciferin Figure 3A
(Miska and Geiger 1987). To test bioluminescence as a read-out for
unconventional secretion, an expression strain producing FLuc-
Cts1 was generated in the background of the octuple protease
deletion strain (AB33P8Δ/FLuc-Cts1). Similarly, a FLuc-Jps1
expressing
strain
was
generated
(AB33P8Δ/FLuc-Jps1)
to
evaluate the effect of the alternative carrier (Figure 3B). AB33
producing intracellular luciferase (FLucCyt) was used as a positive
control in all assays (Müntjes et al., 2020). Monitoring of
proliferation revealed that growth speed was slightly reduced in
AB33P8Δ/FLuc-Jps1 with a doubling time of 3.5 h, compared to the
progenitor strain AB33P8Δ and AB33P8Δ/FLuc-Cts1 showing
doubling
times
of
3 h
in
the
exponential
growth
phase
(Supplementary
Figure
S2). The
slight
reduction
might
eventually be caused by a minor increase in the number of
abnormal cells growing in clusters in the FLuc-Jps1 expressing
strain (Supplementary Figure 2C). Luciferase assays showed that
intracellular activity was very low in the FLuc-Cts1 expressing strain
compared to the strain producing cytoplasmic FLuc, while levels of
Fluc-Jps1 expressing strains were comparable to the cytoplasmic
control showing significant activity (Figure 3C). Importantly, in
culture supernatants the observed effect was even more pronounced
and extracellular FLuc activity for the strain producing FLuc-Jps1
was about 48-fold higher than activity of FLuc-Cts1 secreting cells
for which no significant difference to the control strain could be
observed (Figure 3D). These results were confirmed in Western
blot analyses. While intracellular levels of FLuc-Cts1 were reduced
in comparison to FLuc-Jps1 which showed an about 1.8-fold higher
signal intensity, only FLuc-Jps1 was detectable in supernatants
(Figure 3E; Supplementary Figure S3). This demonstrates that
not only expression of FLuc-Cts1 was impaired but also detectable
Cts1 based secretion was absent. The reason for the differential
performance of the Cts1 and Jps1 fusions with FLuc remains DISCUSSION (A) Bi-specific anti SARS-CoV2 spike-RBD
sybodies sy#15 and sy#68 (Walter et al., 2020) were tagged with a 10x His tag and fused to either Cts1 (top) or Jps1 (bottom) via a TEV protease cleavage site and an
HA-tag. (B) Detection reaction for ELISA: Colorless 10-acetyl-3,7-dihydrophenoxazine (ADHP) is converted by horseradish peroxidase (HRP) using H2O2 to resurofin, a
purple substance that emits strong fluorescence (excitation 570 nm, emission 600 nm). (C) Representative Western blot analyses of Sy68/15-Cts1 and Sy68/15-
Jps1. Secreted protein was enriched from the supernatant by TCA precipitation. Intracellular protein levels were visualized by cell extracts. Western blots show 1 ml of
precipitated supernatants (TCA) and 10 μg cell extracts (CE). Full length protein signals indicated by arrows, degradation bands with rhombi. (D) ELISA of cell extracts:
1 µg of RBD was immobilized per well. 1 µg BSA dealt as a negative control. Baseline was established by a well coated with RBD and only treated with anti-HA and anti-
mouse-HRP. Serial dilutions of U. maydis cell extracts (30 ng, 60 ng, 250 ng per well) were applied in technical triplicates both to RBD and BSA coated wells. Detection
was carried out with the before mentioned anti-HA-mouse and anti-mouse-HRP antibodies. Three biological replicates are shown. Error bars indicate standard deviation
of biological replicates. (E) ELISA of protein purified from supernatants: ELISA wells were coated, and reactions detected as described in (D). Culture supernatants
containing sybody-fusion proteins were subjected to Nickel2+-NTA IMAC and subsequently concentrated up to 10-fold. Serial dilutions of supernatants (1-fold, 5-fold,
10-fold concentrated supernatant) were mixed with blocking solution and added to ELISA wells in technical triplicates. Three biological replicates are shown. Error bars
indicate standard deviation for biological replicates. hydrolytic enzymes are secreted with very high titers compared to
heterologous targets (Nevalainen and Peterson 2014). In our
system, similar to the previously used carrier chitinase Cts1,
Jps1 was fused to the C-terminus of heterologous target
proteins to mediate their export via the fragmentation zone. Of note, one exception identified during this study was the
reporter enzyme LacZ: Here, a LacZ-Cts1 fusion is functional
and unconventionally secreted (Reindl et al., 2020) while
strains producing the respective LacZ-Jps1 fusion showed
growth
retardation
and
were
lacking
detectable
LacZ
activity and LacZ-Jps1 protein in the culture supernatant
(results not shown). DISCUSSION Here we successfully evaluated the potential anchoring factor Jps1
as a novel carrier for the export of heterologous proteins by
unconventional secretion in U. maydis. Carrier proteins are
ubiquitously used in fungal protein expression systems based
on conventional secretion (Fleissner and Dersch 2010). This is
mainly due to the observation that homologous proteins like January 2022 | Volume 9 | Article 816335 Frontiers in Cell and Developmental Biology | www.frontiersin.org 5 Philipp et al. New Carrier for Protein Export FIGURE 4 | Export of functional bi-specific Sars-CoV2 sybodies using Jps1 as a carrier for unconventional secretion. (A) Bi-specific anti SARS-CoV2 spike-RBD
sybodies sy#15 and sy#68 (Walter et al., 2020) were tagged with a 10x His tag and fused to either Cts1 (top) or Jps1 (bottom) via a TEV protease cleavage site and an
HA-tag. (B) Detection reaction for ELISA: Colorless 10-acetyl-3,7-dihydrophenoxazine (ADHP) is converted by horseradish peroxidase (HRP) using H2O2 to resurofin, a
purple substance that emits strong fluorescence (excitation 570 nm, emission 600 nm). (C) Representative Western blot analyses of Sy68/15-Cts1 and Sy68/15-
Jps1. Secreted protein was enriched from the supernatant by TCA precipitation. Intracellular protein levels were visualized by cell extracts. Western blots show 1 ml of
precipitated supernatants (TCA) and 10 μg cell extracts (CE). Full length protein signals indicated by arrows, degradation bands with rhombi. (D) ELISA of cell extracts:
1 µg of RBD was immobilized per well. 1 µg BSA dealt as a negative control. Baseline was established by a well coated with RBD and only treated with anti-HA and anti-
mouse-HRP. Serial dilutions of U. maydis cell extracts (30 ng, 60 ng, 250 ng per well) were applied in technical triplicates both to RBD and BSA coated wells. Detection
was carried out with the before mentioned anti-HA-mouse and anti-mouse-HRP antibodies. Three biological replicates are shown. Error bars indicate standard deviation
of biological replicates. (E) ELISA of protein purified from supernatants: ELISA wells were coated, and reactions detected as described in (D). Culture supernatants
containing sybody-fusion proteins were subjected to Nickel2+-NTA IMAC and subsequently concentrated up to 10-fold. Serial dilutions of supernatants (1-fold, 5-fold,
10-fold concentrated supernatant) were mixed with blocking solution and added to ELISA wells in technical triplicates. Three biological replicates are shown. Error bars
indicate standard deviation for biological replicates. FIGURE 4 | Export of functional bi-specific Sars-CoV2 sybodies using Jps1 as a carrier for unconventional secretion. DISCUSSION We anticipate that this could be related
to the formation of tetramers by LacZ which interfere with Jps1
but not with Cts1 secretion; however, this hypothesis needs to
be verified. Nevertheless, the discovery of a second carrier for
unconventional secretion in U. maydis is a very favorable addition to our expression system (Reindl et al., 2019;
Wierckx et al., 2021): The choice between the two fusion
proteins, Cts1 and Jps1, will greatly enhance the repertoire
of our secretion targets. Jps1 proofed valuable for the export of
proteins that were not secreted at significant levels as Cts1
fusions and showed promising secretion levels for these
targets. This is for example true for the firefly luciferase
FLuc or the bi-specific sybodies that were only secreted
efficiently when fused to Jps1. As a positive side effect, the
FLuc-Jps1 fusion protein is a valuable alternative that allows a
quick and inexpensive quantification of unconventional
secretion via Jps1 in future studies (Wider and Picard 2017;
Branchini et al., 2018). On the contrary, the intrinsic feature of
chitin binding of Cts1 is very attractive as a tool which can be
developed for efficient in situ protein purification from culture
broth (Terfrüchte et al., 2017). Hence, both carriers show January 2022 | Volume 9 | Article 816335 Frontiers in Cell and Developmental Biology | www.frontiersin.org 6 Philipp et al. New Carrier for Protein Export distinct advantages that can be exploited depending on the
actual demands. of a new carrier and export of functional SARS-CoV2
nanobodies we have thus laid a solid foundation for further
exploitation and application of lock-type unconventional
secretion. In line with our results, different carriers show varying
efficiencies in other fungal systems. For example, glycoamylase
or α-amylase have been described as a powerful tool for
heterologous
protein
secretion
in
filamentous
fungi
like
Aspergilli (Ward et al., 1990; Nakajima et al., 2006). Similarly,
the choice of the conventional signal peptide for efficient entry
into the endoplasmic reticulum has been described as a key
factor for improving conventional secretion (Xu et al., 2018;
Wang et al., 2020). While existence of a signal peptide remains
elusive for lock-type unconventional secretion (Stock et al.,
2012), it is conceivable that other unconventionally secreted
proteins are still to be discovered that might constitute even
more powerful carriers. Currently, we do not have a precise
idea on why Jps1 mediates export of heterologous proteins
more effectively than Cts1. Molecular Biology Methods Molecular Biology Methods
All plasmids (pUMa/pUx vectors) generated in this study were
obtained using standard molecular biology methods established
for U. maydis including Golden Gate and Gibson cloning
(Brachmann et al., 2004; Gibson 2011; Gibson et al., 2009;
Terfrüchte
et
al.,
2014). All
plasmids
were
verified
by
restriction analysis and sequencing. Oligonucleotides applied
for cloning are listed in Table 1. Genomic DNA of U. maydis
strain UM521 was used as template for PCR reactions. The
genomic sequence for this strain is stored at the EnsemblFungi
database
(EnsemblFungi). The
generation
of
plasmids
pUMa3329_Δupp1_Pcrg-eGfp-Tnos-natR, pUMa2113_pRabX1-
Poma_gus-SHH-cts1,
pUMa2240_Ip_Poma-his-αGfpllama-ha-
Cts1-CbxR and pUMa3771_Δupp3_Potef_FLuc_NatR has been
described previously (resulting strains, see Table 2). For the
generation of pUMa3012_Ip_Poma_Gus-SHH-Jps1_CbxR the
jps1 gene (umag_03776) was amplified from genomic DNA
using primers oMB372 and oMB373 with AscI and ApaI
hydrolyzation
sites. Subsequently,
the
backbone
of
pUMa2113_Ip_Poma_Gus-SHH-Cts1_CbxR
was
used
for
restriction ligation cloning and jps1 was inserted into the
backbone instead of cts1. pUMa4131_Ip_Poma_FLuc-SHH-
Cts1_CbxR was generated by amplification of the U. maydis
dicodon-optimized
P. pyralis
fluc
gene
from
pUMa3771_Δupp3_Potef_FLuc_NatR
using
oAB297
and
oAB298
with
BamHI
and
SfiI
hydrolyzation
sites. pUMa2113_Ip_Poma_Gus-SHH-Cts1_CbxR
was
then
hydrolyzed with BamHI and SfiI and fluc was inserted into the
backbone instead of gus via restriction/ligation cloning. A
restriction/ligation
cloning
approach
was
applied
for
pUMa4566_Ip_Poma_FLuc-SHH-Jps1_CbxR. jps1 was excised
from pUMa3012_Ip_Poma_Gus-SHH-Jps1_CbxR using AscI
and ApaI and inserted into pUMa4131_Ip_Poma_FLuc-SHH-
Cts1_CbxR,
also
hydrolyzed
with
AscI
and
ApaI. pUx1_Ip_Poma-Sy#68-his-Sy#15-ha-Cts1-CbxR was generated
by amplification of genes sy#68 and sy#15 (Walter et al., 2020)
from a synthetic gBlock (Integrated DNA Technology, Coralville,
Iowa, United States) using primers oAB908 and oAB909 for sy#15
adding BamHI and SpeI hydrolyzation sites and oCD234 and
oCD235 for sy#68 with complementary overhangs for Gibson
cloning. Subsequently, pUMa2240_Ip_Poma-his-αGfpllama-ha-
Cts1-CbxR (Terfrüchte et al., 2017) was hydrolyzed with BamHI
and SpeI and gene sy#15 was inserted via restriction ligation
cloning,
replacing
αgfpllama
and
thereby
generating
pUMa4678. pUMa4678 was then hydrolyzed with BamHI and
the sequence encoding for sy#68 was inserted via Gibson cloning
(Gibson et al., 2009), generating pUx1. For the generation of
pUx8 jps1 was excised from pUMa3012 using AscI and ApaI and
inserted into the AscI and ApaI hydrolyzed backbone of pUx1. The
successful
synthesis
and
functional
export
of
nanobodies directed against the RBD of the surface spike
protein of the SARS-CoV2 virus is a timely new addition to
the repertoire of secreted targets. DISCUSSION Further studies on the molecular
roles of Jps1 during Cts1 secretion might resolve this question
in the future. Notably, unconventional secretion was also
observed for septation factor Don3 (Aschenbroich et al.,
2019) which may thus serve as such alternative carrier. However, Gus activity levels of unconventionally secreted
Gus-Don3 are minute compared to Gus-Cts1, suggesting
that
it
does
not
constitute
a
promising
alternative
(Aschenbroich et al., 2019). Hence, it is important to
further study the mechanism of lock-type secretion and in
particular, to identify further players that localize to the
fragmentation zone for export during cytokinesis (Reindl
et al., 2019; Wierckx et al., 2021). Strain Generation U. maydis strains used in this study were obtained by homologous
recombination yielding genetically stable strains (Bösch et al., 2016)
(Table 2). For genomic integrations at the ip locus, integrative
plasmids were used (Stock et al., 2012). These plasmids contained
the ipr allele, promoting carboxin resistance. For integration, plasmids
were linearized within the ipr allele to allow for homologous
recombination with the ips locus. For transformation, integrative
plasmids were hydrolyzed within the ipr locus using the restriction
endonuclease SspI, resulting in a linear DNA fragment. For genetic
modifications in other loci, plasmids with about 1 kb flanking regions
and a resistance cassette were generated (Brachmann et al., 2004;
Terfrüchte et al., 2014). For transformation, the insertion cassette was
excised from the plasmid backbone using SspI or SwaI (Terfrüchte
et al., 2014). For all genetic manipulations, U. maydis protoplasts were
transformed
with
linear
DNA
fragments
for
homologous
recombination. All strains were verified by Southern blot analysis
(Southern 1974). For in locus modifications the flanking regions were
amplified as probes. For ip insertions, the probe was obtained by PCR
using the primer combination oMF502/oMF503 and the template
pUMa260 (Keon et al., 1991; Brachmann et al., 2004). Primer
sequences are listed in Table 1. Quantification of Unconventional Secretion
by Western blot analysis U. maydis strains were grown at 28°C in complete medium
supplemented with 1% (w/v) glucose (CM-glc) or with 1% (w/v)
arabinose (CM-ara) if not described differently (Holliday 1974;
Tsukuda et al., 1988). Solid media were supplemented with 2%
(w/v) agar agar. Growth phenotypes were evaluated using the
BioLector
microbioreactor
(m2p-labs,
Baesweiler,
Germany)
(Funke
et
al.,
2010). MTP-R48-B(OH)
round
plates
were
inoculated
with
1.5 ml
culture
per
well
and
incubated
at
1,000 rpm at 28°C. Backscatter light with a gain of 25 or 20 was
used to determine biomass. Gus-Cts1 and Gus-Jps1 secretion was analyzed by trichloroacetic
acid (TCA) precipitation of culture broths. 1 ml of cell-free
supernatants of cultures grown in Verduyn medium (55.5 mM
Glucose, 74.7 mM NH4Cl, 0.81 mM MgSO4×7H2O, 0.036 mM
FeSO4×7H2O,
36.7 mM
KH2PO4,
100 mM
MES
pH
6.5,
0.051 mM EDTA, 0.025 mM ZnSO4×7H2O, 0.041 mM CaCl2,
0.016 mM H3bBO3, 6.7 µM MnCl2×2H2O, 2.3 µM CoCl2×6H2O,
1.9 µM CuSO4×5H2O, 1.9 µM Na2MoO4×2H2O, 0.6 µM KI) to
an OD600 of 3 were chilled on ice and mixed with 400 µl 50%
(v/v) TCA solution and incubated on ice at 4°C overnight. Subsequently, precipitated protein pellets were harvested by
centrifugation at 11,000 x g at 4°C for 30 min. Supernatants
were discarded and pellets were washed with 300 µl of-20°C
acetone followed by centrifugation at 11,000 × g at 4°C for Molecular Biology Methods Determination of Extracellular Cts1 Activity
Extracellular
Cts1
activity
was
analyzed
using
4-
methylumbelliferyl β-D cellobioside (MUC, Sigma-Aldrich,
Billerica, MA, United States) as a substrate (Koepke et al.,
2011). Whole cell cultures were mixed 3:7 with KHM Buffer
(110 mM CH3CO2K, 20 mM HEPES, 2 mM MgCl2, 2 mM MUC)
in black 96 well plates. Relative fluorescence units were
determined
using
a
plate
reader
(Tecan,
Männedorf,
Switzerland) by fluorescence measurement at 28°C for 100 min
every 2 min (360 nm excitation and 450 nm emission, gain 100). Determination of Extracellular Cts1 Activity
Extracellular
Cts1
activity
was
analyzed
using
4-
methylumbelliferyl β-D cellobioside (MUC, Sigma-Aldrich,
Billerica, MA, United States) as a substrate (Koepke et al.,
2011). Whole cell cultures were mixed 3:7 with KHM Buffer
(110 mM CH3CO2K, 20 mM HEPES, 2 mM MgCl2, 2 mM MUC)
in black 96 well plates. Relative fluorescence units were
determined
using
a
plate
reader
(Tecan,
Männedorf,
Switzerland) by fluorescence measurement at 28°C for 100 min
every 2 min (360 nm excitation and 450 nm emission, gain 100). Quantification of Unconventional Secretion
Using Luciferase Reporter g
Extra- and intracellular luciferase activity was determined using
D-luciferin (Biosynth Carbosynth, Compton, United Kingdom). Cell-free supernatants or whole cell cultures in CM medium were
mixed 8:2 with luciferin substrate mix (20 mM tricine, 2.67 mM
MgSO4×7H2O, 0.1 mM EDTA×2 H2O, 33.3 mM DTT, 0.524 mM
ATP, 0.269 mM acetyl-CoA, 0.467 mM D-luciferin, 5 mM
NaOH, 0.264 mM MgCO3×5H2O) in white 96-well plates. Relative photon count (RPC) was determined using a Mithras
LB 940 plate reader (Berthold technologies, Bad Wildbad,
Germany) for 20 min with measurements every 20 s. Molecular Biology Methods The current pandemics
situation underpinned that it is important to develop novel
methodology for quick, specific, and sensitive detection and
treatment of viral infections in the future. On the one hand
nanobodies
are
potent
proteins
for
antigen
detection
(Muyldermans 2013) and thus very promising tools in the
context of SARS-CoV2 detection. On the other hand,
antibody-based
pharmaceuticals
like
Casirivimab
and
Imdevimab are already used to treat COVID-19 infection
(Sun and Ho 2020). Therefore, besides application in virus
diagnostics, nanobodies directed against SARS-CoV2 could
potentially even become novel pharmaceutical targets for
therapeutic approaches (Dubey et al., 2020). The unique
system of unconventional secretion in U. maydis now
offers new possibilities for nanobody production without
the risk of undesired modifications by N-glycosylation
(Stock et al., 2012). This would eliminate the necessity to
humanize
llama
derived
nanobodies
for
safe
use
as
pharmaceuticals
to
avoid
allergic
reaction
in
patients
(Vincke et al., 2009; Dong et al., 2020). To achieve this,
both the unconventional secretion system and specifically
the production and application of nanobodies via this
system have to be optimized, for example by further
multimerization to increase valency and affinity (Wichgers
Schreur et al., 2020; Koenig et al., 2021). By the establishment January 2022 | Volume 9 | Article 816335 Frontiers in Cell and Developmental Biology | www.frontiersin.org 7 New Carrier for Protein Export Philipp et al. TABLE 1 | DNA oligonucleotides used in this study. Designation
Nucleotide sequence (59- 39)
oMB372_jps1_fw
TTAGGCGCGCCATGCCAGGCATCTCC
oMB373_jps1_rev
TTAGGGCCCTTAGGATTCCGCATCGATTGGGG
oMF502_ip_fw
ACGACGTTGTAAAACGACGGCCAG
oMF503_ip_rev
TTCACACAGGAAACAGCTATGACC
oAB297_fluc_fw
AAATTGGATCCATGGAGGACGCCAAGAACATCAAG
oAB298_fluc_rev
AATAGGCCGCGTTGGCCACGGCGATCTTGCCACCCTT
oAB908_sy#15_fw
ATATAGGATCCATGGCGGCCCATCACCACCATCACC
ACCATCACCACCATCATATGCAGGTGCAGCTCG
oAB909_sy#15_rev
ATATAACTAGTCGAGACGGTGACCTGGGTGC
oCD234_sy#68_fw
CTACCTTACTCTATCAGGATCATGCAGGTGCAGCTC
GTCG
oCD235_sy#68_rev
GGTGATGGGCCGCCATGGATCCCGAGACGGTGACCT
GGGTGC TABLE 1 | DNA oligonucleotides used in this study. methylumbelliferyl β-D galactopyranoside (MUG, bioWORLD,
Dublin, OH, United States)) (Koepke et al., 2011; Stock et al.,
2012; Stock et al., 2016). Cell-free culture supernatants were
mixed
1:1
with
2×
Gus
assay
buffer
(10 mM
sodium
phosphate buffer pH 7.0, 28 µM β-mercaptoethanol, 0.8 mM
EDTA, 0.0042% (v/v) lauroyl-sarcosin, 0.004% (v/v) Triton X-
100, 2 mM MUG, 0.2 mg/ml (w/v) BSA) in black 96-well plates. Relative fluorescence units (RFUs) were determined using a plate
reader (Tecan, Männedorf, Switzerland) for 100 min at 28°C with
measurements every 5 min (excitation/emission wavelengths:
365/465 nm, gain 60). For quantification of conversion of
MUG to the fluorescent product 4-methylumbelliferone (MU),
a calibration curve was determined using 0, 1, 5, 10, 25, 50, 100,
200 µM MU. Frontiers in Cell and Developmental Biology | www.frontiersin.org SDS PAGE and Western Blot Analysis SDS PAGE and Western Blot Analysis
To verify protein production and secretion in cell extracts and
supernatants, respectively, Western Blot analysis was used. 20 ml
cultures were grown to an OD600 of 1.0 and harvested at 1,500
×
g
for
5 min
in
centrifugation
tubes. Until
further
preparation, pellets were stored at −20°C. For preparation of
cell extracts, cell pellets were resuspended in 1 ml cell extract
lysis buffer (100 mM sodium phosphate buffer pH 8.0, 10 mM
Tris/HCl pH 8.0, 8 M urea, 1 mM DTT, 1 mM PMSF, 2.5 mM
benzamidine, 1 mM pepstatin A, 2× complete protease
inhibitor cocktail (Roche, Sigma/Aldrich, Billerica, MA,
United States) and cells were crushed by agitation with
glass
beads
at
2,500 rpm
for
12 min
at
4°C. After
centrifugation
(11,000
×
g
for
30 min
at
4°C),
the Quantification of Unconventional Secretion
Using the Gus Reporter bPlasmids generated in our working group are integrated in a plasmid collection and termed pUMa, or pUx plus a number as 4-digit number as identifier. TABLE 2 | (Continued) U. maydis strains used in this study. Strains
Relevant
genotype/Resistance
Strain collection
no. (UMaa)
Plasmids
transformed/Resistanceb
Manipulated locus
(umag gene
identifier)
Progenitor
(UMaa)
References
ipS(Pomafluc:shh:jps1)ipR CbxR
AB33P8ΔSy#68/
#15-Cts1
a2 PnarbW2bE1 PhleoR
Ux1
pUx1
ip
2413
this study
FRT10(um04641Δ:hyg)
FRT11(um03947Δ)
FRT6(um03975Δ)
FRT5(um04400Δ)
FRT3(um11908Δ)
FRT2(um00064Δ)
FRTwt[um02178Δ)
FRT1(um04926Δ) HygR
ipS(Pomaantirbdsybody#68:his:
antirbdsybody#15:ha:cts1)ipR
CbxR
AB33P8ΔSy#68/
#15-Jps1
a2 PnarbW2bE1 PhleoR
Ux8
pUx8
ip
2413
this study
FRT10(um04641Δ:hyg)
FRT11(um03947Δ)
FRT6(um03975Δ)
FRT5(um04400Δ)
FRT3(um11908Δ)
FRT2(um00064Δ)
FRTwt[um02178Δ)
FRT1(um04926Δ) HygR
ipS(Pomaantirbdsybody#68:his:
antirbdsybody#15:ha:jps1)ipR
CbxR
aInternal strain collection numbers (UMa/Ux codes). bPlasmids generated in our working group are integrated in a plasmid collection and termed pUMa, or pUx plus a number as 4-digit number as identifier. TABLE 2 | (Continued) U. maydis strains used in this study. Plasmids
transformed/Resistanceb 2413
this study this study Internal strain collection numbers (UMa/Ux codes). Plasmids generated in our working group are integrated in a plasmid c supernatant
was
separated
from
cell
debris
and
was
transferred to a fresh reaction tube. Protein concentration
was determined by Bradford assay (BioRad, Hercules, CA,
United States) (Bradford 1976) and 10 µg total protein was
used for SDS-PAGE. SDS-PAGE was conducted using 10% (w/
v) acrylamide gels. Subsequently, proteins were transferred to
methanol activated PVDF membranes using semi-dry Western
blotting. SHH-tagged Gus-Cts1 was detected using a primary
anti-HA (1:3,000, Millipore/Sigma, Billerica, United States). An anti-mouse IgG-horseradish peroxidase (HRP) conjugate
(1:3,000 Promega, Fitchburg, United States) was used as
secondary antibody. HRP activity was detected using the
Amersham ™ECL ™Prime Western Blotting Detection
Reagent
(GE
Healthcare,
Chalfont
St
Giles,
United
Kingdom)
and
a
LAS4000
chemiluminescence
imager (GE Healthcare Life Sciences, Freiburg, Germany). 20 min twice. Pellets were dried at room temperature and
resuspended in Laemmli buffer containing 0.12 M NaOH. Resuspended pellets were denatured at 95°C for 10 min and
then subjected to SDS-PAGE and Western blot analysis. To
determine
protein
concentration
obtained
by
TCA
precipitation a standard ladder of 50, 100, 200 and 500 ng
of Multiple Tag protein (GenScript Biotech, Piscataway, NJ,
United States) was loaded onto the SDS-PAGE next to
obtained samples. Western blot signals were quantified
using image studio lite version 5.2 (Li-Cor Biosciences,
Lincoln,
NE,
United
States)
and
the
standard
curve
obtained by quantification of Multiple Tag protein signals
was used to determine protein concentrations in culture
supernatants. Quantification of Unconventional Secretion
Using the Gus Reporter g
p
Extracellular
Gus
activity
was
determined
to
quantify
unconventional Cts1 secretion using the specific substrate 4- January 2022 | Volume 9 | Article 816335 Frontiers in Cell and Developmental Biology | www.frontiersin.org 8 New Carrier for Protein Export Philipp et al. TABLE 2 | U. maydis strains used in this study. Strains
Relevant
genotype/Resistance
Strain collection
no. (UMaa)
Plasmids
transformed/Resistanceb
Manipulated locus
(umag gene
identifier)
Progenitor
(UMaa)
References
AB33
a2 PnarbW2bE1
133
pAB33
b
FB2 (55)
Brachmann et al. (2004)
PhleoR
AB33 Gus-Cts1
a2 PnarbW2bE1 PhleoR
1289
pUMa2113/CbxR
ip
133
Sarkari et al. (2014)
ipS(Pomagus:shh:cts1)ipRCbxR
AB33don3Δ/Gus-
Cts1
a2 PnarbW2bE1 PhleoR
1742
pUMa2717/HygR
umag_05543 (don3)
1289
Aschenbroich
et al. (2019)
ipS(Pomagus:shh:cts1)ipR CbxR
umag_don3Δ_HygR
AB33don3Δ
a2 PnarbW2bE1 PhleoR
2028
pUMa2717/HygR
umag_05543 (don3)
133
Aschenbroich
et al. (2019)
umag_don3Δ_HygR
AB33don3Δ/
Pcrgdon3-gfp/Gus-
Cts1
a2 PnarbW2bE1 PhleoR
2302
pUMa3330/NatR
umag_02178 (upp1)
1742
Aschenbroich
et al. (2019)
ipS(Pomagus:shh:cts1)ipR CbxR
umag_don3Δ_HygR
upp1:(Pcrgdon3:gfp) NatR
AB33P8ΔGus-Cts1
a2 PnarbW2bE1 PhleoR
2418
pUMa2113
Ip
2413
Terfrüchte et al. (2018)
FRT10(um04641Δ:hyg)
FRT11(um03947Δ)
FRT6(um03975Δ)
FRT5(um04400Δ)
FRT3(um11908Δ)
FRT2(um00064Δ)
FRTwt[um02178Δ)
FRT1(um04926Δ) HygR
ipS(Pomagus:shh:cts1)ipR CbxR
AB33don3Δ/Gus-
Jps1
a2 PnarbW2bE1 PhleoR
2734
pUMa3012
Ip
2028
This study
ipS(Pomagus:shh:cts1)ipR CbxR
umag_don3Δ_HygR
AB33don3Δ/
Pcrgdon3-gfp/Gus-
Jps1
a2 PnarbW2bE1 PhleoR
2776
pUMa3330/NatR
umag_02178 (upp1)
2734
This study
ipS(Pomagus:shh:cts1)ipR CbxR
umag_don3Δ_HygR
upp1:(Pcrgdon3:gfp) NatR
AB33P8ΔGus-Jps1
a2 PnarbW2bE1 PhleoR
2900
pUMa3012
Ip
2413
this study
FRT10(um04641Δ:hyg)
FRT11(um03947Δ)
FRT6(um03975Δ)
FRT5(um04400Δ)
FRT3(um11908Δ)
FRT2(um00064Δ)
FRTwt[um02178Δ)
FRT1(um04926Δ) HygR
ipS(Pomagus:shh:jps1)ipR CbxR
AB33P8Δ FLuc-Cts1
a2 PnarbW2bE1 PhleoR
3151
pUMa4131
Ip
2413
this study
FRT10(um04641Δ:hyg)
FRT11(um03947Δ)
FRT6(um03975Δ)
FRT5(um04400Δ)
FRT3(um11908Δ)
FRT2(um00064Δ)
FRTwt[um02178Δ)
FRT1(um04926Δ) HygR
ipS(Pomafluc:shh:cts1)ipR CbxR
AB33P8Δ FLuc-Jps1
a2 PnarbW2bE1 PhleoR
3214
pUMa4566
ip
this study
FRT10(um04641Δ:hyg)
FRT11(um03947Δ)
FRT6(um03975Δ)
FRT5(um04400Δ)
FRT3(um11908Δ)
FRT2(um00064Δ)
FRTwt[um02178Δ)
FRT1(um04926Δ) HygR TABLE 2 | U. maydis strains used in this study. 2413
this study this study (Continued on following page)
January 2022 | Volume 9 | Article 816335 January 2022 | Volume 9 | Article 816335 New Carrier for Protein Export New Carrier for Protein Export Philipp et al. TABLE 2 | (Continued) U. maydis strains used in this study. Strains
Relevant
genotype/Resistance
Strain collection
no. (UMaa)
Plasmids
transformed/Resistanceb
Manipulated locus
(umag gene
identifier)
Progenitor
(UMaa)
References
ipS(Pomafluc:shh:jps1)ipR CbxR
AB33P8ΔSy#68/
#15-Cts1
a2 PnarbW2bE1 PhleoR
Ux1
pUx1
ip
2413
this study
FRT10(um04641Δ:hyg)
FRT11(um03947Δ)
FRT6(um03975Δ)
FRT5(um04400Δ)
FRT3(um11908Δ)
FRT2(um00064Δ)
FRTwt[um02178Δ)
FRT1(um04926Δ) HygR
ipS(Pomaantirbdsybody#68:his:
antirbdsybody#15:ha:cts1)ipR
CbxR
AB33P8ΔSy#68/
#15-Jps1
a2 PnarbW2bE1 PhleoR
Ux8
pUx8
ip
2413
this study
FRT10(um04641Δ:hyg)
FRT11(um03947Δ)
FRT6(um03975Δ)
FRT5(um04400Δ)
FRT3(um11908Δ)
FRT2(um00064Δ)
FRTwt[um02178Δ)
FRT1(um04926Δ) HygR
ipS(Pomaantirbdsybody#68:his:
antirbdsybody#15:ha:jps1)ipR
CbxR
aInternal strain collection numbers (UMa/Ux codes). nzyme-Linked Immunosorbent Assay Enzyme-Linked Immunosorbent Assay
For detection of nanobody binding activity protein adsorbing 384-
well
microtiter
plates
(Nunc®
Maxisorp™,
ThermoFisher
Scientific, Waltham, MA, United States) were used. Wells were
coated with 1 µg commercially available Sars-CoV2 spike-RBD-
domain protein (GenScript Biotech, Piscataway, NJ, United States). 1 µg BSA per well dealt as negative control (NEB, Ipswich, MA,
United States). Samples were applied in a final volume of 100 µl
coating buffer (100 mM Tris-HCL pH 8, 150 mM NaCl, 1 mM
EDTA) per well at 4°C for at least 16 h. Blocking was conducted for
at least 4 h at 4°C with 5% (w/v) skimmed milk in coating buffer. Subsequently, 5% (w/v) skimmed milk in PBS was added to defined
protein amounts of nanobody samples from cell extracts or
purified from culture supernatants and respective controls. 100 µl of sample was added to wells coated with Sars-CoV2
spike-RBD and BSA. The plate was incubated with samples and
controls over night at 4°C. After 3x PBS-T (PBS supplemented with
0.05% (v/v) Tween-20, 100 µl per well) washing, a mouse anti-HA
antibody (Millipore/Sigma, Billerica, United States) 1: 5,000 diluted
in PBS supplemented with skimmed milk (5% w/v) was added
(100 µl per well) and incubated for 2 h at room temperature. Then
wells were washed again three times with PBS-T (100 µl per well)
and incubated with an anti-mouse IgG-horseradish peroxidase
(HRP) conjugate (Promega, Fitchburg, United States) (50 µl per
well) for 1 h at room temperature [1:5,000 in PBS supplemented
with skimmed milk (5% w/v)]. Subsequently wells were washed
three times with PBS-T and three times with PBS and incubated
with Quanta Red™enhanced chemifluorescent HRP substrate (50:
50:1, 50 µl per well) (ThermoFisher Scientific, Waltham, MA,
United States) at room temperature for 15 min. The reaction
was stopped with 10 µl per well Quanta RedTM stop solution
and fluorescence readout was performed at 570 nm excitation and
600 nm emission using an Infinite M200 plate reader (Tecan,
Männedorf, Switzerland). ACKNOWLEDGMENTS We are thankful to B. Axler for excellent technical support of the
project. We gratefully acknowledge support in microscopic
imaging and analysis by N. Heßler. FUNDING KH
was
supported
by
the
CLIB-Competence
Center
Biotechnology
(CKB)
funded
by
the
European
Regional
Development Fund ERDF (34. EFRE-0300096). KS and MR
received funding from the Deutsche Forschungsgemeinschaft
(DFG,
German
Research
Foundation)—Projektnummer
267205415—SFB 1208. KH
was
supported
by
the
CLIB-Competence
Center
Biotechnology
(CKB)
funded
by
the
European
Regional
Development Fund ERDF (34. EFRE-0300096). KS and MR
received funding from the Deutsche Forschungsgemeinschaft
(DFG,
German
Research
Foundation)—Projektnummer
267205415—SFB 1208. Microscopic Analyses The Supplementary Material for this article can be found online at:
https://www.frontiersin.org/articles/10.3389/fcell.2021.816335/
full#supplementary-material Microscopic analyses were performed with immobilized early-log
phase budding cells on agarose patches (3% w/v f. c.) using a Biophys. Acta (Bba)—Proteins Proteomics 1867, 140154, 2019 . Epub 2018/
10/15. doi:10.1016/j.bbapap.2018.10.007 DATA AVAILABILITY STATEMENT The original contributions presented in the study are included in
the article/Supplementary Material, further inquiries can be
directed to the corresponding author. AUTHOR CONTRIBUTIONS MP, KPH and MR designed the experiments. MP and KPH
conducted the experiments. KS, KM and MF supervised the
project. KS, MP and KM prepared the manuscript with input
from all co-authors. MP and KH prepared figures and tables. IMAC Purification of Supernatants For the purification of recombinant unconventionally secreted
protein from U. maydis, cells were grown in CM-glucose (1%
w/v) medium buffered with 0.05 M MES pH 6.5.200 ml of culture
supernatants were harvested at and OD600 of 0.8 by centrifugation at
5,000 × g for 3 min. Harvested supernatants were chilled to 4°C and
treated with a protease inhibitor tablet of cOmplete protease inhibitor
(Roche, Sigma/Aldrich, Billerica, MA, United States). 2 ml of
Nickel2+-NTA matrix was equilibrated with 50 ml lysis buffer
(10 mM imidazole 50 mM NaH2PO4, 300 mM NaCl, pH 8.0). 22 ml of 10 times concentrated lysis buffer were added to the
supernatants and subsequently Nickel2+-NTA matrix was added January 2022 | Volume 9 | Article 816335 Frontiers in Cell and Developmental Biology | www.frontiersin.org 10 New Carrier for Protein Export Philipp et al. to the supernatant. The mixture was batched by gentle stirring on a
magnetic stirrer at 4°C for 1 h. Following batching supernatant flow-
through was discarded via a PD-10 column. Matrix was collected in
the PD-10 column during the process. Collected matrix was washed
with 50 ml of wash buffer (lysis buffer, 20 mM Imidazole) and protein
was eluted with 2 ml elution buffer (lysis buffer, 250 mM imidazole). In the last step supernatants were concentrated via Amicon Ultra 50 k
0.5 ml centrifugal filter devices (Merck Millipore, Burlington,
Massachusetts, United States) and the buffer exchanged to PBS
(137 mM
NaCl,
2.7
mM
KCl,
10 mM
Na2HPO4,
1.8 mM
KH2PO4, pH 7.2) and applied for ELISA. wide-field microscope setup from Zeiss (Oberkochen, Germany)
Axio Imager M1 equipped with a Spot Pursuit CCD camera
(Diagnostic Instruments, Sterling Heights, United States) and the
objective lenses Plan Neofluar (×40, NA 1.3), Plan Neofluar (63×,
NA 1.25) and Plan Neofluar (100×, NA 1.4). The microscopic
system was controlled by the software MetaMorph (Molecular
Devices, version 7, Sunnyvale, United States). Image processing
including rotating and cropping of images, scaling of brightness,
contrast and fluorescence intensities as well as insertion of scale
bars was performed with MetaMorph. Arrangement and
visualization were performed with Canvas 12 (ACD Systems). Biophys. Acta (Bba)—Proteins Proteomics 1867, 140154, 2019 . Epub 2018/
10/15. doi:10.1016/j.bbapap.2018.10.007 REFERENCES (2020). Emerging Antibody-Based Therapeutics against SARS-CoV-
2 during the Global Pandemic. Antibody Ther. 3, 246–256. doi:10.1093/abt/tbaa025 Kaplon, H., and Reichert, J. M. (2021). Antibodies to Watch in 2021. mAbs 13,
1860476. doi:10.1080/19420862.2020.1860476 Terfrüchte, M., Joehnk, B., Fajardo-Somera, R., Braus, G. H., Riquelme, M.,
Schipper, K., et al. (2014). Establishing a Versatile Golden Gate Cloning
System for Genetic Engineering in Fungi. Fungal Genet. Biol. 62, 1–10. doi:10.1016/j.fgb.2013.10.012 Keon, J. P. R., White, G. A., and Hargreaves, J. A. (1991). Isolation,
Characterization and Sequence of a Gene Conferring Resistance to the
Systemic Fungicide Carboxin from the maize Smut Pathogen, Ustilago
Maydis. Curr. Genet. 19, 475–481. Epub 1991/06/01. doi:10.1007/bf00312739 Terfrüchte, M., Reindl, M., Jankowski, S., Sarkari, P., Feldbrügge, M., and Schipper,
K. (2017). Applying Unconventional Secretion in Ustilago Maydis for the
export of Functional Nanobodies. Ijms 18, 937. doi:10.3390/ijms18050937 Koenig, P.-A., Das, H., Liu, H., Kümmerer, B. M., Gohr, F. N., Jenster, L.-M., et al. (2021). Structure-guided Multivalent Nanobodies Block SARS-CoV-2 Infection
and Suppress Mutational Escape. Science 371, 371, 2021 . Epub 2021/01/14. doi:10.1126/science.abe6230 Terfrüchte, M., Wewetzer, S., Sarkari, P., Stollewerk, D., Franz-Wachtel, M.,
Macek,
B.,
et
al. (2018). Tackling
Destructive
Proteolysis
of
Unconventionally
Secreted
Heterologous
Proteins
in
Ustilago
Maydis. J. Biotechnol. 284,
28437–28451. Epub
2018/08/01. doi:10.1016/
j.jbiotec.2018.07.035 Koepke, J., Kaffarnik, F., Haag, C., Zarnack, K., Luscombe, N. M., König, J., et al. (2011). The RNA-Binding Protein Rrm4 Is Essential for Efficient Secretion of
Endochitinase Cts1. Mol. Cell Proteomics 10, M111 011213, 2011 . Epub 2011/
08/03. doi:10.1074/mcp.M111.011213 Tsukuda, T., Carleton, S., Fotheringham, S., and Holloman, W. K. (1988). Isolation
and Characterization of an Autonomously Replicating Sequence from Ustilago
Maydis. Mol. Cel. Biol. 8, 3703–3709. Epub 1988/09/01. doi:10.1128/
mcb.8.9.3703-3709.1988 Langner, T., Öztürk, M., Hartmann, S., Cord-Landwehr, S., Moerschbacher, B.,
Walton, J. D., et al. (2015). Chitinases Are Essential for Cell Separation in
Ustilago Maydis. Eukaryot. Cel 14, 846–857. Epub 2015/05/03. doi:10.1128/
EC.00022-15 Vincke, C., Loris, R., Saerens, D., Martinez-Rodriguez, S., Muyldermans, S., and
Conrath, K. (2009). General Strategy to Humanize a Camelid Single-Domain
Antibody and Identification of a Universal Humanized Nanobody Scaffold. Vincke, C., Loris, R., Saerens, D., Martinez-Rodriguez, S., Muyldermans, S., and
Conrath, K. (2009). General Strategy to Humanize a Camelid Single-Domain
Antibody and Identification of a Universal Humanized Nanobody Scaffold. J. Biol. Chem. 284,
2843273–2843284. Epub
2008/11/18. doi:10.1074/
jbc.M806889200 Miska, W., and Geiger, R. (1987). Synthesis and Characterization of Luciferin
Derivatives for Use in Bioluminescence Enhanced Enzyme Immunoassays. New
Ultrasensitive Detect. REFERENCES Bösch, K., Frantzeskakis, L., Vraneš, M., Kämper, J., Schipper, K., and Göhre, V. (2016). Genetic Manipulation of the Plant Pathogen Ustilago Maydis to Study
Fungal
Biology and Plant Microbe
Interactions. JoVE 30, 20160930. doi:10.3791/54522 Aschenbroich, J., Hussnaetter, K. P., Stoffels, P., Langner, T., Zander, S., Sandrock,
B., et al. (2019). The Germinal centre Kinase Don3 Is Crucial for
Unconventional Secretion of Chitinase Cts1 in Ustilago Maydis. Biochim. January 2022 | Volume 9 | Article 816335 Frontiers in Cell and Developmental Biology | www.frontiersin.org 11 Philipp et al. New Carrier for Protein Export Brachmann, A., König, J., Julius, C., and Feldbrügge, M. (2004). A Reverse Genetic
Approach for Generating Gene Replacement Mutants in Ustilago Maydis. Mol. Genet. Genomics 272, 216–226. Epub 2004/08/19. doi:10.1007/s00438-004-
1047-z Model Microorganism Ustilago Maydis. Front. Microbiol. 11, 1384, 2020 . Epub 2020/07/17. doi:10.3389/fmicb.2020.01384 Muyldermans, S. (2013). Nanobodies: Natural Single-Domain Antibodies. Annu. Rev. Biochem. 82, 775–797. Epub 2013/03/19. doi:10.1146/annurev-biochem-063011-
092449 Bradford, M. M. (1976). A Rapid and Sensitive Method for the Quantitation of
Microgram Quantities of Protein Utilizing the Principle of Protein-Dye
Binding. Anal. Biochem. 72 (72), 248–254. Epub 1976/05/07. doi:10.1006/
abio.1976.9999 Nakajima, K.-i., Asakura, T., Maruyama, J.-i., Morita, Y., Oike, H., Shimizu-Ibuka,
A., et al. (2006). Extracellular Production of Neoculin, a Sweet-Tasting
Heterodimeric Protein with Taste-Modifying Activity, by Aspergillus oryzae. Appl. Environ. Microbiol. 72, 3716–3723. doi:10.1128/aem.72.5.3716-3723.2006 Branchini, B. R., Southworth, T. L., Fontaine, D. M., Kohrt, D., Florentine, C. M.,
and Grossel, M. J. (2018). A Firefly Luciferase Dual Color Bioluminescence
Reporter Assay Using Two Substrates to Simultaneously Monitor Two Gene
Expression Events. Sci. Rep. 8 (8), 5990, 2018 . Epub 2018/04/18. doi:10.1038/
s41598-018-24278-2 Nevalainen, H., and Peterson, R. (2014). Making Recombinant Proteins in
Filamentous Fungi- Are We Expecting Too Much? Front. Microbiol. 5, 75,
2014 . Epub 2014/03/01. doi:10.3389/fmicb.2014.00075 Nicaud, J.-M., Mackman, N., and Holland, I. B. (1986). Current Status of Secretion
of Foreign Proteins by Microorganisms. J. Biotechnol. 3, 255–270. doi:10.1016/
0168-1656(86)90008-8 Dong, J., Huang, B., Jia, Z., Wang, B., Gallolu Kankanamalage, S., Titong, A., et al. (2020). Development of Multi-specific Humanized Llama Antibodies Blocking
SARS-CoV-2/ace2 Interaction with High Affinity and Avidity. Emerging
Microbes
&
Infections
9,
1034–1036. Epub
2020/05/15. doi:10.1080/
22221751.2020.1768806 Nickel, W. (2010). Pathways of Unconventional Protein Secretion. Curr. Opin. Biotechnol. 21, 621–626. Epub 2010/07/20. doi:10.1016/j.copbio.2010.06.004 Rabouille, C. (2017). Pathways of Unconventional Protein Secretion. Trends Cel
Biol. 27, 230–240. Epub 2016/12/19. doi:10.1016/j.tcb.2016.11.007 Dubey, A., Dahiya, S., Rouse, B. T., and Sehrawat, S. (2020). REFERENCES Perspective: Reducing
SARS-CoV2 Infectivity and its Associated Immunopathology. Front. Immunol. 11, 11. doi:10.3389/fimmu.2020.581076 Reichert, J. M. (2015). Antibodies to Watch in 2015. mAbs 7, 1–8. doi:10.4161/
19420862.2015.988944 Flaschel, E., and Friehs, K. (1993). Improvement of Downstream Processing of
Recombinant Proteins by Means of Genetic Engineering Methods. Biotechnol. Adv. 11, 31–77. doi:10.1016/0734-9750(93)90409-G Reindl, M., Hänsch, S., Weidtkamp-Peters, S., and Schipper, K. (2019). A Potential
Lock-type Mechanism for Unconventional Secretion in Fungi. Ijms 20, 460. doi:10.3390/ijms20030460 Fleissner, A., and Dersch, P. (2010). Expression and export: Recombinant Protein
Production Systems for Aspergillus. Appl. Microbiol. Biotechnol. 87, 1255–1270. Epub 2010/06/10. doi:10.1007/s00253-010-2672-6 Reindl, M., Stock, J., Hussnaetter, K. P., Genc, A., Brachmann, A., and Schipper, K. (2020). A Novel Factor Essential for Unconventional Secretion of Chitinase
Cts1. Front. Microbiol. 11, 1529, 2020 . Epub 2020.02.07. doi:10.1101/
2020.02.07.93861310.3389/fmicb.2020.01529 Funke, M., Buchenauer, A., Schnakenberg, U., Mokwa, W., Diederichs, S., Mertens, A.,
et al. (2010). Microfluidic Biolector-Microfluidic Bioprocess Control in Microtiter
Plates. Biotechnol. Bioeng. 107, 497–505. Epub 2010/06/03. doi:10.1002/bit.22825 Sarkari, P., Reindl, M., Stock, J., Müller, O., Kahmann, R., Feldbrügge, M., et al. (2014). Improved Expression of Single-Chain Antibodies in Ustilago Maydis. J. Biotechnol. 191, 165–175. doi:10.1016/j.jbiotec.2014.06.028 Gibson, D. G. (2011). Enzymatic Assembly of Overlapping DNA Fragments. Methods Enzymol. 498, 349–361. Epub 2011/05/24. doi:10.1016/B978-0-12-
385120-8.00015-2 Southern, E. M. (1974). Detection of Specific Sequences Among DNA Fragments
Separated by Gel Electrophoresis. J. Mol. Biol. 98, 503–517. doi:10.1016/s0022-
2836(75)80083-0 Gibson, D. G., Young, L., Chuang, R.-Y., Venter, J. C., Hutchison, C. A., 3rd, and
Smith, H. O. (2009). Enzymatic Assembly of DNA Molecules up to Several
Hundred Kilobases. Nat. Methods 6, 343–345. Epub 2009/04/14. doi:10.1038/
nmeth.1318 Stock, J., Sarkari, P., Kreibich, S., Brefort, T., Feldbrügge, M., and Schipper, K. (2012). Applying Unconventional Secretion of the Endochitinase Cts1 to export
Heterologous Proteins in Ustilago Maydis. J. Biotechnol. 161, 16180–16191. Epub 2012/03/27. doi:10.1016/j.jbiotec.2012.03.004 Holliday, R. (1974). “Ustilago Maydis,” in Handbook of Genetics (New York, NY:
Plenum Press), 575–595. doi:10.1007/978-1-4899-1710-2_31 Epub 2012/03/27. doi:10.1016/j.jbiotec.2012.03.004 Stock, J., Terfrüchte, M., and Schipper, K. (2016). A Reporter System to Study
Unconventional Secretion of Proteins Avoiding N-Glycosylation in Ustilago
Maydis. Unconventional Protein Secretion: Methods Protoc. Springer Protoc. 1459, 149–160. doi:10.1007/978-1-4939-3804-9_10 Hussnaetter, K. P., Philipp, M., Müntjes, K., Feldbrügge, M., and Schipper, K. (2021). Controlling Unconventional Secretion for Production of Heterologous
Proteins in Ustilago Maydis through Transcriptional Regulation and Chemical
Inhibition of the Kinase Don3. JoF 7, 179, 2021 . Epub 2021/04/04. doi:10.3390/
jof7030179 Sun, Y., and Ho, M. REFERENCES Syst. Enzyme Immunoassays, 25:23–30. doi:10.1515/
cclm.1987.25.1.23 J. Biol. Chem. 284,
2843273–2843284. Epub
2008/11/18. doi:10.1074/
jbc.M806889200 Viotti, C. (2016). ER to Golgi-dependent Protein Secretion: The Conventional
Pathway. Methods Mol. Biol. 1459, 3–29. Epub 2016/09/26. doi:10.1007/978-1-
4939-3804-9_1 Müntjes, K., Philipp, M., Hüsemann, L., Heucken, N., Weidtkamp-Peters, S.,
Schipper, K., et al. (2020). Establishing Polycistronic Expression in the Frontiers in Cell and Developmental Biology | www.frontiersin.org January 2022 | Volume 9 | Article 816335 12 Philipp et al. New Carrier for Protein Export Walsh, G. (2018). Biopharmaceutical Benchmarks 2018. Nat. Biotechnol. 36,
1136–1145. doi:10.1038/nbt.4305 Factories. Essays
Biochem. Jul
65
(65),
365–379. Epub
2021/04/17. doi:10.1042/EBC20200141 Walter, J. D., Hutter, C. A. J., Zimmermann, I., EgloffWyss, P., Sorgenfrei, M.,
Hürlimann, L. M., et al. (2020). Sybodies Targeting the SARS-CoV-2 Receptor-
Binding Domain. bioRxiv. doi:10.1101/2020.04.16.045419 Xu, Y., Wang, Y.-h., Liu, T.-q., Zhang, H., Zhang, H., and Li, J. (2018). The GlaA
Signal Peptide Substantially Increases the Expression and Secretion of α-
galactosidase in Aspergillus niger. Biotechnol. Lett. 40, 949–955. doi:10.1007/
s10529-018-2540-5 Wang, Q., Zhong, C., and Xiao, H. (2020). Genetic Engineering of Filamentous
Fungi for Efficient Protein Expression and Secretion. Front. Bioeng. Biotechnol. 8, 293, 2020 . Epub 2020/04/24. doi:10.3389/fbioe.2020.00293 Conflict of Interest: The authors declare that the research was conducted in the
absence of any commercial or financial relationships that could be construed as a
potential conflict of interest. Ward, M., Wilson, L. J., Kodama, K. H., Rey, M. W., and Berka, R. M. (1990). Improved Production of Chymosin in Aspergillus by Expression as a
Glucoamylase-Chymosin Fusion. Nat. Biotechnol. 8, 435–440. Epub 1990/
05/01. doi:10.1038/nbt0590-435 Publisher’s Note: All claims expressed in this article are solely those of the authors
and do not necessarily represent those of their affiliated organizations, or those of
the publisher, the editors, and the reviewers. Any product that may be evaluated in
this article, or claim that may be made by its manufacturer, is not guaranteed or
endorsed by the publisher. Weinzierl, G., Leveleki, L., Hassel, A., Kost, G., Wanner, G., and Bolker, M. (2002). Regulation of Cell Separation in the Dimorphic Fungus Ustilago
Maydis. Mol. Microbiol. 45, 219–231. Epub 2002/07/09. doi:10.1046/j.1365-
2958.2002.03010.x Wichgers Schreur, P. J., van de Water, S., Harmsen, M., Bermúdez-Méndez, E., Drabek,
D., Grosveld, F., et al. (2020). Multimeric Single-Domain Antibody Complexes
Protect against Bunyavirus Infections. eLife 9, e52716. doi:10.7554/eLife.52716 Copyright © 2022 Philipp, Hussnaetter, Reindl, Müntjes, Feldbrügge and Schipper. Frontiers in Cell and Developmental Biology | www.frontiersin.org January 2022 | Volume 9 | Article 816335 REFERENCES This is an open-access article distributed under the terms of the Creative Commons
Attribution License (CC BY). The use, distribution or reproduction in other forums is
permitted, provided the original author(s) and the copyright owner(s) are credited
and that the original publication in this journal is cited, in accordance with accepted
academic practice. No use, distribution or reproduction is permitted which does not
comply with these terms. Wider, D., and Picard, D. (2017). Secreted Dual Reporter Assay with Gaussia
Luciferase and the Red Fluorescent Protein mCherry. PLoS One 12, e0189403,
2017 . Epub 2017/12/09. doi:10.1371/journal.pone.0189403 Wierckx, N., Miebach, K., Ihling, N., Hussnaetter, K. P., Büchs, J., and Schipper, K. (2021). Perspectives for the Application of Ustilaginaceae as Biotech Cell January 2022 | Volume 9 | Article 816335 Frontiers in Cell and Developmental Biology | www.frontiersin.org 13
|
https://openalex.org/W4365606991
|
https://zenodo.org/records/7831060/files/Xudoyberdiyev%20Anvar%20Megli%20o%E2%80%99gli.pdf
|
Swahili
| null |
JADIDCHILIK HARAKATINING TUB MOHIYATI HAMDA JADIDLAR HAYOTIDAGI QORA SAHIFALAR
|
Zenodo (CERN European Organization for Nuclear Research)
| 2,023
|
cc-by
| 3,000
|
https://doi.org/10.5281/zenodo.7831060 https://doi.org/10.5281/zenodo.7831060 Xudoyberdiyev Anvar Megli o’gli
anvarxudoyberdiyev258@gmail.com
O’zbekiston Milliy universiteti Tarix fakulteti
Tarix yo’nalishi 1-bosqich talabasi Annotatsiya. XIX asr oxiri XX asr boshlarida Rossiya mustamlakasi hisoblangan O’rto osiyo, Sibr,
Kavkaz hududlarda boshlangan madaniy marifiy o’zgarishlari ziyoliy qatlam orasida
Jadidchilik harakati nomi bilan keng yoyildi.Maqolamizda dastlab Jadidchilik harakatininig
yuzaga kelish sabablari va g’oyaviy qarama-qarshiliklari, xalqni qoloqlikdan hamda diniy
xurofotdan ozod etish,ma’naviy-marifiy isloxotlar orqali jamiyatni yuksaltirish va siyosiy
birlikka chorlash kabi g’oyaviy qarashlari haqida atroflicha fikr yuritsak, maqolamizning
ikkinchi qismida Jadidchilik harakatining yorqin namoyondalarining hayoti va ularning
mustabid tuzimga qarshi olib borgan milliy ozodlik harakatlarini, shuningdek yakunda
sodir bo’lga oktabrning mudhish voqealarini maqolamiz sahifalari ketma-ketligida yoritib
beramiz. Kalit so’zlar. Jadidchilik, Maorif, Yangi usul , Tarbiai Atvol, Mustabid, Jaholatga qarshi kurash,
Panturkizm, Qora sahifalar, Asri orzu. Abstract. The cultural and educational changes that began in the regions of Central Asia,
Siberia, and the Caucasus, which were considered Russian colonies at the end of the 19th
century and the beginning of the 20th century, spread widely among the intellectuals under
the name of Jadidchilik movement.In our article, we will firstly think about the reasons, for
the emergence of the Jadidism movement and its , its idealogical opposition, its idealogical
views, such as liberating the people from backwardness and religious prejudice, improving
society through spiritual and educational reforms, and calling for political unity.We will
cower the life of its bright representatives and their national liberation movements against
the established regime , as well the terrible events of October that happened in the end ,in
the sequence of pages of our article. Key words. Key words. 936 Publishing centre of Finland International Journal of Education, Social Science & Humanities. Finland Academic Research Science Publishers
ISSN: 2945-4492 (online) | (SJIF) = 7.502 Impact factor
Volume-11| Issue-4| 2023 Published: |22-04-2023| International Journal of Education, Social Science & Humanities. Finland Academic Research Science Publishers
ISSN: 2945-4492 (online) | (SJIF) = 7.502 Impact factor
Volume-11| Issue-4| 2023 Published: |22-04-2023| Jadidism, Educatio, New method, Tarbiai Atvol, Mustabid system, Strugle against
ignorance, Pan-Turkism, Blac pages, Century dream. Jadidism, Educatio, New method, Tarbiai Atvol, Mustabid system, Strugle against
ignorance, Pan-Turkism, Blac pages, Century dream. g
,
,
p g ,
y
Maqolamiz
avvalida
“Jadid”
atamsi
kelib
chiqish
tarixiga
nazar
tashlasak.Dastlab Turkiyada Sulton Salim III {1739-1802} hukmronligi davrida
“Jadid” atamasi siyosiy tuzimga nisbatan qo’llanilgan. Abu Bakr Ratib
bildirishnomalarida keltirishicha u yerda ko’rgan idora tizimini dastlab “Nizomi
Jadid” deb nomlagan.Yevropaga bu terminning kirib kelishi 1789-yildagi buyuk
Fransuz inqilobiga borib taqaladi.Ushbu jamiyatda qurilgan siyosiy tuzim
“Fransiya nizomi Jadid” nomi bilan tarixiy manbalarda keltirilgan.XIX asr oxiri va
XXasr boshlarida insoniyat sivilizatsiyasida tub burilish davri bo’ldi.Davlatlarning
keyingi siyosiy taraqqiyoti yoki qaramlik davrini jahon urishlari belgilab berdi.Bu
davrda bir jamiyat yoki xalqning butunlay yod jamiyat ta’sirida qolishi turmush
hayotda balki insonlar falsafasida inqilobiy fikrlar shakillanishiga asos bo’ldi. Aynan shu davrda Markaziy Osiyoda boshqa mintaqalarida shakillangani singari
o’zgacha fikrlovchi ziyoliylar qatlami shakillanishi yangi Jadidchilik deb
nomlangan oqim vujudga kelishiga zamin yaratdi.Jadidchilik yoki Jadidizm (
arabcha jadid yangi ) Turkiston, Kavkaz, Qirim xalqlar hayotida muhim ahamiyat
kasb etgan siyosiy-ijtimoiy tuzum desak mubolag’a bo’lmaydi. Abstract. O’ng oqim o’z oldiga
xon hokimiyatini saqlab qolgan holda iqtisodiy sohada o’zgarishlar amalga oshirish
orqali erkin bozor munosabatlariga keng yo’l ochishni maqsad qilib qo’ygan.Xiva
jadidchiligining so’l oqimi esa mayda sarmoyadorlar va xalqning turli tabaqa
vakillarini birlashtirgan bo’lib ularga qozikalon Bobooxun Salimov rahbarlik
qilgan.Ularning asosiy birdan bir maqsadi xon hokimiyatiga qarshi kurash bu
orqali yangi usul maktablar tarmog’ini kengytirishni o’z oldiga dasturilamal
sifatida qo’ygan edi. Jadidchilik harakati namoyondalaridan bo’lgan Mahmudxo’ja
Behbudiy, Sadriddin Ayniy, Fayzulla Xo’jayev kabilar mustabid tuzumga qarshi
kurashishning murosa yo’lini tanlagan bo’lsa, Abdurauf Fitrat, Abdulhamid
Cho’lpon, Munavvarqori Abdurashidxonov kabilar modernizatsiyaning radikal
yo’lini tanlashgan. Jadidchilikning poydevoriga tamal toshi qo’yilishi yangi usul
maktablari tashkil etilishi edi. Maqolamizning davomida maorif tuzimi yuksalishi
hamda yangi usul maktablari tashkil topishi haqida atroflicha fikr yuritamiz. Rossiyaga
qaram
bo’lgan
musulmon
xalqlar
orasida
jadidlar
harakatini
rivojlantirishda Istanbulda ta’lim olgan , siyosat va jamoat arbobi Ismoil Gaspiral
(1851-1914) katta rol o’ynaydi.Sharq tillari, madaniyat, mintaliteti bilan tanish
bo’lgan Gaspiral osiyoning musulmon mamlakatlarini madaniy saviyasini
oshirishga, mamlakatni modernizatsiyasi qilishga to’sqinlik qilayotgan to’siqlar XIX
asrning
ikkinchi
yarmida
Jadidchilik
harakatining
turkistonda
tarqalishning mafkuraviy yo’nalishi uch g’oyaviy frontdan iborat bo’lgan.Ular
Turkiston,Buxoro, Xiva jadidchilik kabi frontlar edi. .Turkiston jadidchiligining
ijtimoiy asosini ziyolilar tashkil qilgan .Ular Rossiya mustabid hokimiyatiga qarshi
kurashning oldingi saflarida turishdi.Turkiston dastlab muxtor so’ngra mustaqil
davlat
bo’lishini
yo’qlab
chiqdilar
.Buxoro
jadidchiligining
asosini
taraqqiyparvarlar, hunarmandlardan iborat bo’lgan. XX asr boshlarida Buxoro
jamiyati 2 gruhga : Ikrom domla rahbarligidagi taraqqiyparvarlar, Mulla
Abdurazzoq boshchiligida qadimiylarga bo’lingan edi. Ular turkistondagi ijtimoiy
siyosiy vaziyatdan kelib chiqqan holda hukumat oldiga soliqlarni kamaytirish
talabi bilan chiqdi. Xiva jadidchiligi asosiy ikkita oqimdan tashkil topgan.O’ng
oqimni harakatlantiruvchi kuch rivojlaniyotgan savdo-sanoat korxonalari egalari
hamda
yirik
boylarning
vakillari
hisoblangan. Bu
oqimga
Xiva
xoni
Asfandiyorxonning bosh vaziri Islomxo’ja boshchilik qilgan. O’ng oqim o’z oldiga
xon hokimiyatini saqlab qolgan holda iqtisodiy sohada o’zgarishlar amalga oshirish
orqali erkin bozor munosabatlariga keng yo’l ochishni maqsad qilib qo’ygan.Xiva
jadidchiligining so’l oqimi esa mayda sarmoyadorlar va xalqning turli tabaqa
vakillarini birlashtirgan bo’lib ularga qozikalon Bobooxun Salimov rahbarlik
qilgan.Ularning asosiy birdan bir maqsadi xon hokimiyatiga qarshi kurash bu
orqali yangi usul maktablar tarmog’ini kengytirishni o’z oldiga dasturilamal
sifatida qo’ygan edi. Jadidchilik harakati namoyondalaridan bo’lgan Mahmudxo’ja
Behbudiy, Sadriddin Ayniy, Fayzulla Xo’jayev kabilar mustabid tuzumga qarshi
kurashishning murosa yo’lini tanlagan bo’lsa, Abdurauf Fitrat, Abdulhamid
Cho’lpon, Munavvarqori Abdurashidxonov kabilar modernizatsiyaning radikal
yo’lini tanlashgan. Jadidchilikning poydevoriga tamal toshi qo’yilishi yangi usul
maktablari tashkil etilishi edi. Maqolamizning davomida maorif tuzimi yuksalishi
hamda yangi usul maktablari tashkil topishi haqida atroflicha fikr yuritamiz. Abstract. XIX asr oxirida
mustamlaka zulmi tufayli iqtisodiy jihatdan inqiroz holatiga tushib qolgan,
Turkistonni jaholat iskanjasidan olib chiqishga bel bog’lagan ziyoliylar birinchi
navbatda Chor Rossiyaning siyosiy zulmiga qarshi kurashishni afzal bilishdi.Bu
borada Jadidchilik harakati katta rol o’ynadi.Jadidchilik harakatining asosiy
maqsad va tamoillari shunda ediki qoloq ijtimoiy tuzumni bartaraf etish
mamlakat aholisi orasida ma’rifatni keng yoyish va madaniy islohatlar o’tkazish,
shuningdek milliy mustaqillik g’oyalarini hayotga tadbiq etishni o’z oldiga maqsad
qilib qo’ygan edi. Turkistonda jadidchilik g’oyalari XIX asr 90-yillarida keng
yoyildi .XX asr 30-yillarida o’lka ijtimoiy va siyosiy hayotida muhum rol o’ynadi
.Jadidchilik umummilliy harakat sifatida quydagi g’oyalar va yo’nalishlarni ilgari
surdi. *Turkistonni o’rta asrlarga xos bo’lgan qoloqlik va dini xurofotdan ozod etish
va shariatni isloh qilish . *Turkistonni o’rta asrlarga xos bo’lgan qoloqlik va dini xurofotdan ozod etish
va shariatni isloh qilish . *Turkiston muxtoriyat hukumatini barpo etish, Buxoro va Xivada demokratik
respublika tuzimini o’rnatish orqali ozod va farovon jamiyat qurish *Milliy pul birligi , Milliy qo’shin tuzish va Milliy davlatchilik tuzumini
shakillantirish. 937 Publishing centre of Finland International Journal of Education, Social Science & Humanities. Finland Academic Research Science Publishers
ISSN: 2945-4492 (online) | (SJIF) = 7.502 Impact factor
Volume-11| Issue-4| 2023 Published: |22-04-2023| International Journal of Education, Social Science & Humanities. Finland Academic Research Science Publishers International Journal of Education, Social Science & Humanities. Finland Academic Research Science Publishers
ISSN: 2945-4492 (online) | (SJIF) = 7.502 Impact factor
Volume-11| Issue-4| 2023 Published: |22-04-2023| *Yangi usul maktablar tarmog’ini kengaytirish,qobiliyatli yoshlarni chet elga
ta’lim olish uchun yuborish. *Yangi usul maktablar tarmog’ini kengaytirish,qobiliyatli yoshlarni chet elga
ta’lim olish uchun yuborish. *Turli marifiy teatr truppalarini tuzish, gazeta va jurnallar chop etish bu orqali
xalqning ma’naviy ongini bir pog’ona yuksaltirish XIX
asrning
ikkinchi
yarmida
Jadidchilik
harakatining
turkistonda
tarqalishning mafkuraviy yo’nalishi uch g’oyaviy frontdan iborat bo’lgan.Ular
Turkiston,Buxoro, Xiva jadidchilik kabi frontlar edi. .Turkiston jadidchiligining
ijtimoiy asosini ziyolilar tashkil qilgan .Ular Rossiya mustabid hokimiyatiga qarshi
kurashning oldingi saflarida turishdi.Turkiston dastlab muxtor so’ngra mustaqil
davlat
bo’lishini
yo’qlab
chiqdilar
.Buxoro
jadidchiligining
asosini
taraqqiyparvarlar, hunarmandlardan iborat bo’lgan. XX asr boshlarida Buxoro
jamiyati 2 gruhga : Ikrom domla rahbarligidagi taraqqiyparvarlar, Mulla
Abdurazzoq boshchiligida qadimiylarga bo’lingan edi. Ular turkistondagi ijtimoiy
siyosiy vaziyatdan kelib chiqqan holda hukumat oldiga soliqlarni kamaytirish
talabi bilan chiqdi. Xiva jadidchiligi asosiy ikkita oqimdan tashkil topgan.O’ng
oqimni harakatlantiruvchi kuch rivojlaniyotgan savdo-sanoat korxonalari egalari
hamda
yirik
boylarning
vakillari
hisoblangan. Bu
oqimga
Xiva
xoni
Asfandiyorxonning bosh vaziri Islomxo’ja boshchilik qilgan. Publishing centre of Finland Abstract. Rossiyaga
qaram
bo’lgan
musulmon
xalqlar
orasida
jadidlar
harakatini
rivojlantirishda Istanbulda ta’lim olgan , siyosat va jamoat arbobi Ismoil Gaspiral
(1851-1914) katta rol o’ynaydi.Sharq tillari, madaniyat, mintaliteti bilan tanish
bo’lgan Gaspiral osiyoning musulmon mamlakatlarini madaniy saviyasini
oshirishga, mamlakatni modernizatsiyasi qilishga to’sqinlik qilayotgan to’siqlar 938 Publishing centre of Finland International Journal of Education, Social Science & Humanities. Finland Academic Research Science Publishers
ISSN: 2945-4492 (online) | (SJIF) = 7.502 Impact factor
Volume-11| Issue-4| 2023 Published: |22-04-2023| International Journal of Education, Social Science & Humanities. Finland Academic Research Science Publishers
ISSN: 2945-4492 (online) | (SJIF) = 7.502 Impact factor
Volume-11| Issue-4| 2023 Published: |22-04-2023| bartaraf qilishga intildi. Buxoroga kelib Sulton Ahad bilan uchrashdi va unga
o’qitishning yangi usulidan foydalangan holda islom bilan bir qatorda dunyoviy
fanlarni o’qitish tizimiga asoslangan maktablar tashkil qilish taklifini beradi Lekin
an’anaviy mafkura tarafdori bo’lgan Sulton Ahad bu fikrni qo’llab quvvatlamaydi. Ismoil Gaspiral modernizatsiya qilishning dastlabki bosqichini ta’lim tizimini isloh
qilishdan boshladi.Chunki mustabid tuzumga bolta urish uchun avvalo xalqning
g’oyaviy mafkurasini shakillantirish zarur edi. 1884-yilda jadid maktabini tashkil
qilib, 40 kunda 12ta bolaning savodini chiqardi. Uning bu tuzimi “usuli savtiya”
yani “yangi usul” nomi bilan shuxrat qozondi. 1888-Yilda “Rahbari muallim ,
muallimlar yo’ldoshi” asarini yaratdi. Ommaviy axborot vositalari orqali milliy
ozodlik g’oyalarini keng yoyish uchun foydalandi. Bu yo’lda “Tarjimon” gazetasi
yashirin qurol vazifasini bajardi. Ushbu gazeta (1883-1914) yillar oralig’ida mavjud
bo’lgan. Undan tashqari Munavvarqori rahbarligida “Xurshid”, Abdulla Avloniy
muharirligida “ Shuxrat”, Bektemirov tashabbusi bilan “Osiyo”gazetalari nashir
qilina boshlandi.1913-1915-yillar oralig’ida matbuotchilikning yangi to’lqini
shakillana boshladi. Xususan “Sadoi Turkiston”, “Sadoi Farg’ona”, “Xurriyat”,
“Ulug’ turkiston” kabi ommaviy axborot vositalari keng ommaga taqdim etildi. I.Gaspiral 1893 yilda Samarqand, Toshkent, Buxoro shaharlarida yangicha tuzimni
keng yoyishga harakat qildi .Buxorodalik paytida Gaspiral Abdulahadni jadid
maktabini ochishga ko’ndiradi. Bu maktab “Muzaffariya” nomi bilan ataldi. 1898-
yilda To’qmoqda jadid maktablari ochildi. Shamsiddin domla boshchiligida
Andijonda 1898-yilda , 1901-yilda Qo’qonda Salohiddin domla, Toshkentda
Munavvarqori
Abdurashidxonov,
Samarqandda
Abduqodir
Shakuriy
boshchiligida ilk jadid maktablari ochildi. 1908-yilda Buxoroda Abdulvohid
Munzim tomonidan dastlabki tojik tilidagi yangi usul maktabini tashkil qilindi. Lekin bu yerda mutassiblik g’oyalar tarafdorlari shu qadar ko’p ediki buning
oqibatida 1909-yilda Munzimning tashkil qilgan maorif maskani yopiladi.1911-1912
yillarda Buxoroda yangi usul maktablari soni 57 tani tashkil qilgan.Ularning
orasida eng mashhurlari Mirkomil Burxonov, Usmonxoja Po’lat xojayev, Xolidxo’ja
Mehrilarniki(1913) bo’lgan.Buxoroda joylashgan Mulla Vafoning maktabida
o’qitish rus tiliga asoslangan edi. Publishing centre of Finland Abstract. 1911-yilga kelib Turkistondagi yangi usul
maktablar soni 63 ta bo’lib unda taxsil oladigan talabalarning soni 4106 tani tashkil
etgan,
yozma
malumotlarga
asoslansak
hududlar
kesimida
maktablar
taqsimlanishi quydagicha:Farg’ona viloyatida 36ta, Yettisuvda 12 tasi, 5 tasi
Samarqandda Toshkentning o’zida 24ta yangi usul tipidagi maktablar mavjud
bo’lgan. Munavvarqori Abdurashidxonovning1927-yil toshkent okrug maorif 939 Publishing centre of Finland International Journal of Education, Social Science & Humanities. Finland Academic Research Science Publishers
ISSN: 2945-4492 (online) | (SJIF) = 7.502 Impact factor
Volume-11| Issue-4| 2023 Published: |22-04-2023| International Journal of Education, Social Science & Humanities. Finland Academic Research Science Publishers
ISSN: 2945-4492 (online) | (SJIF) = 7.502 Impact factor
Volume-11| Issue-4| 2023 Published: |22-04-2023| xodimlari qurultoyida so’zlagan nutqida shunday takidlagan: “Jadid maktabi
tashkil qilg’onlar ham eski madrasa va qorixonalarda yetishib chiqqan ziyolilar
edi.Ular yolg’iz Boxchasaroyda chiqadurg’on Gasprinskiy gazetalarini o’qirdilar va
shu
orqali
Turkiston
milliy
mafkurasini
shakillantirishga
bel
bog’lagan
edilar,hamda Jadidlar oqimini vujudga keltirdilar”[ 1 ] deb so’zini yakunlaydi. Jadidlar taxsil oliyotgan avlod ongiga milliy vatanparvarlik ruhini singdirish
maqsadida bir qator darsliklar yaratishdi.Jumladan Saidrasul Saidazizov “Ustodi
avval” (1902), Behbudiy “Risolai asbobi savod” (1904), Munavvarqori “Adibi
avval”, “Adibi soniy” (1907), Avloniy “Birinchi muallim” (1910), “Ikkinchi
muallim” kabi asarlar taxsil oliyotgan talabalarni siyosiy qarashlarini bir qadar
yuksalishiga zamin yaratdi. Jadidchilar yangi usul maktablari sonini tez suratlarda
ko’paytirish uchun qilgan harakati bejiz ketmadi. 1917 yilda Turkistonda ularning
soni 100 taga yetdi.Ammo jamiyatda tez fursatda yangicha fikrlaydigan qatlam
shakillanishiga Rossiya ichki ishlar vazirligi jim qarab tura olmas edi. Jadidlarning
usul maktablari tashkil qilib , gazetalar chiqarishlari panislomizmning mintaqa
bo’ylab targ’ib qilish sifatida baholandi va hokimiyat bu borada qarshilik
ko’rsatishga harakat qila boshladi[ 2]. Jadidlarning keyingi maqsadlaridan biri iqtidorli yoshlarni chet el oliy o’quv
yurtlarida taxsil olish uchun jo’natishdan iborat edi. Shu maqsadda 1909-yilning
18-iyulida “Tarbiyai Atvol” marifiy jamiyatini tashkil qilishdi.Moliyaviy jihatdan
Muhiddin Mansurov , Sadri Ziyo kabi sarmoyadorlar jamiyatini qo’llab
quvvatlaganlar.Jamiyat asosini Hamidxo’ja Mehriy, Sadriddin Ayni, Ahmadjon
Abdusaidov tashkil etgan. Ushbu jamiyat (1908-1914) yillar oralig’ida mavjud
bo’lgan. Dastlab 1911-yilda 15nafar yosh kadirni chet elga taxsil olish uchun
jo’natgan bo’lsa ,bu raqamlar 1912- yilda 30tani tashkil qilgan[3]. Turkiston Jadidchilik oqimini harakatlantiruvchi kuch sifatida Mahmudxo’ja
Behbudiy, Abduqodir Shakuriy, Sadriddin Ayniy, Abdulhamid Cho’lpon,
Munavvarqori Abdurashidxonov, Abdurauf Fitrat, Is’hoqxon Ibrat kabi yurt
fidoilari ko’z oldimizda gavdalanadi. Xususan o’rta osiyo Jadidlar otasi
hisoblangan
Mahmudxo’ja
Behbudiy(1875-1919)
Uning
tashabbusi
o’laroq
Samarqandda yangi usul maktablari ochildi. Publishing centre of Finland Abstract. 1909 yilda islom tarixiga oid asar
yaratdi.Mamlakat yoshlarini saviyali ilmi asarlar bilan taminlash maqsadida (1908)
yilda “ Behbudiya” kutubxonasi nomi bilan mashhur qiroatxona tashkil qildi [4]. Bu esa talabalarning g’arb va sharq ilmi manbalaridan bohabar bo’lishi uchun
yaratilgan eng katta imkoniyat edi. Xalqning ma’naviy ongini yuksaltirishda
matbuotning o’rni yuqori ekanligini anglagan holda “Behbudiya” nomi bilan 940 Publishing centre of Finland International Journal of Education, Social Science & Humanities. Finland Academic Research Science Publishers
ISSN: 2945-4492 (online) | (SJIF) = 7.502 Impact factor
Volume-11| Issue-4| 2023 Published: |22-04-2023| International Journal of Education, Social Science & Humanities. Finland Academic Research Science Publishers
ISSN: 2945-4492 (online) | (SJIF) = 7.502 Impact factor
Volume-11| Issue-4| 2023 Published: |22-04-2023| ataladigan xususiy nashriyot tashkil qiladi, shuningdek “oyna” jurnali (1913)
nashir etilishida jonbozlik ko’rsatdi. .Behbudiy (1904-1909) yillarda “Risolai
jug’rofiai umroniya”, “Risolai jug’rofiya Rusiy”, “Kitob ul atvol’’, “Muxtasar
tarixi islom” kabi yuksak vatanparvarlik g’oyalari bilan yo’g’rilgan asarlarni yozib
qoldirgan. Abdurauf Fitrat (1886-1938) o’rta osiyo jadidchiligining taniqli vakili
shuningdek birinchi o’zbek prafessori[5].Fitrat shaxsiyati Jadid marifatparvarlik
harakati shakillanishida asosiy o’rinni egallaydi.Jadidchilik milliy marifatparvarlik
harakatining tarafdori. Inqilobga qadar o’rta osiyoni Rossiyadan ozod qilinish
harakatlarida
jonbozlik
ko’rsatgan
.1909-yilda
Istanbulda
nashr
etilgan
“Munozara” asari mazmunida Buxoro amirligi boshqararuv tizimida islohat
o’tkazish hamda usuli jadid deb nomlangan maorif tizimi asoslari haqida so’z
yuritgan.1906-1907 yillarda qizlar ucun Urganchda maktab ochilishi tashabbuskori
bo’ldi.Shuningdek do’sti Munzim bilan hamkorlikda 70 nafar yosh turkistonliklarni
Germaniyaga tahsil olishga jo’natdi.O’z asarlari mazmunida mustabid tuzimni
qoralab xalqni ilmsizlik iskanjasidan olib chiqishga qaratilgan g’oyalarni ilgari
surganlgi taxsinga sazovordir Munavvarqori
Abdurashidxonov
(1878-1929)
Turkiston
jadidchilik
harakatining yo’lboshchisi XX asr o’zbek teatrchilik san’at, o’sha davrda nashr
etilgan “ Xurshid” gazetasining, shuningdek yangi usul maktablari asoschisi
hisoblanadi. 1910-yilda Toshkentda “Turon” kutubxonasi tashkil etishda oldingi
saflarda turib tashabbus ko’rsatgan. Dastlab 1901-yilda o’z uyida yangi usuli jadid
maktabini tashkil qiladi.Uning “Adibi avval”, “Adibi soniy”(1907), “Tarixi
Anbiyo” kabi qo’llanma darsliklari yangi tashkil etilgan usul maktablarida asosiy
qo’llanmasi sifatida qadrlangan.Munavvarqori 1917-yil fevral oyi voqealaridan
so’ng Turkiston xalq demokratik davlatchilik yaratish g’oyalarini ochiqdan ochiq
ilgari sura boshlaydi.1918-yilda Turk o’chog’i ilmi marifiy jamiyatini tashkil qiladi. Mustabid tuzum zanjirlariga bolta urish uchun yashirin inqilobiy “Milli ittihod va
Milli istiqlol” gruhiga rahbarlik qila boshlagan hamda ozodlik kabi g’oyalarni ilgari
surib xalqni birlikka chorlagan. O’zbek xalqi uchun norasmiy motam kuni sifatida eslanadigan 1938-yilning 4-
oktabr voqealari jadidchilik harakati namoyondalari hayotida qora sahifalarga
muxurlandi.1938-yil 4-oktabrda Toshkent SSSR Oliy sudi Harbiy kollegiyasining
sayyor yig’ilishida 51 nafar mahbus otuvga hukum qilingan.4-16-oktabr oralig’ida
507nafar O’zbekistonlik mahbus ustidan shavqatsiz hukm chiqarildi. Publishing centre of Finland Publishing centre of Finland Abstract. Ular orasida
xalqimizning jaholat botqog’idan olib chiqishga harakat qilgan ammo yakunda 941 Publishing centre of Finland International Journal of Education, Social Science & Humanities. Finland Academic Research Science Publishers
ISSN: 2945-4492 (online) | (SJIF) = 7.502 Impact factor
Volume-11| Issue-4| 2023 Published: |22-04-2023| International Journal of Education, Social Science & Humanities. Finland Academic Research Science Publishers
ISSN: 2945-4492 (online) | (SJIF) = 7.502 Impact factor
Volume-11| Issue-4| 2023 Published: |22-04-2023| otuvga hukm qilingan ziyolilar ham bor edi.Xususan Abdurauf Fitrat , Cho’lpon(
1937-yil xalq dushmani) , Abdulla Qodiriy (1937-yil 31-dekabrda xalq dushmani),
kabi adabiyotimizning yorqin vakillari shuningdek siyosiy arboblarimizdan Rahim
Inog’omov ,Rustam Islomov , Sadulla Tursunxo’jayev kabilar ham qatog’on
qurboni bo’lishdi.Rasmiy malumotlarga ko’ra SSSR oliy sud raisi yurist Aleeksev
tomonidan davlat xafsizlik mayyori Darenik Zaxarovich Apresyanga talabnoma
yo’llab 51 nafar ziyoliy ustidan chiqarilgan qarorga o’zgartirish kiritib zudlik bilan
otuvga
hukim
qiladi[5].Jazo
Yunusobod
tumanidagi
qatilxonada
amalga
oshiriladi.( 1va 2 rasmlarga) etibor bering. Unda Fitrat hamda Qodiriyning oliy sud
qaroriga ko’ra sud jarayoni 5-oktabrda bo’lishi haqidagi hujjat berilgan .Ammo jazo
otriyadi bir kun avval jazoni ijro etish uchun buyruq oladi . Ertasida sud zalida
saxnalashtirilgan sud jarayoni bo’lib o’tadi. Jazo otriyadi tomonidan 4-oktabrda
jazo amalga oshirilayotgan vaqtda aholi o’q ovozini payqamasligi uchun motor
yoqib qo’yilganligi tarixiy ma’lumotlarda uchraydi.Manbalarda keltirilishicha
ushbu kunda shu qadar ko’p mahbuslarning hayotiga zomin bo’lishadiki hatto
avtomatlar o’qlari bunga dosh bermaydi. Cho’lponga kelgan vaqtda o’qlar ham
tugaydi .Cho’lpon bunday qabih kimsalar orasida qolishni istamasligi tabiiy edi, va
yakunda jazo bolta bilan amalga oshiriladi . Davlat xafsizlik katta letinanti
Shishkin tomonida jazo amalga oshirilgani haqida dalolatnoma tayyorlanadi [5]. Yuqoridagi malumotlardan shu ayon bo’ladiki yurtni jaholat botqog’idan olib
chiqish uchun qilgan harakatlari tufayli jazo yana bir oz vaqt kechiktirilganda edi
xalqning istiqlolchilik harakatlari bilan ulanib ketgan bo’lar edi. Ammo ko’p
o’tmay xalq g’aflat uyqusidan uygondi. Milliy ozodlik harakatlari keng yoyila
boshlandi.Podsho
hukumati
millatning
mintalitetiga
urgan
boltasi
o’z
hukumatining ag’darilishiga zamin yaratdi.Jadidchilik harakati xalqimizning
mustabid tuzumdan xalos bo’lishida ,milliy mafkura shakillanishida, shuningdek
xalqimizning diniy va dunyoviy qarashlari bir qadar yuksalishida muhim ahamiyat
kasb etdi. Jadidchilik harakati maorif va siyosiy boshqaruv tizimlarida ham
o’zining ijobiy ta’sirini o’tkazdi. Jadidlar hayotidagi qora saxifalar xalqimiz
xotirasida o’chmas iz qoldirdi.Tarix zarvaraqlarida va xalqimiz ongida xalq
ozodligi yo’lida shahid bo’lgan yurt farzandlari yodi mangu qolajak. 942 Publishing centre of Finland International Journal of Education, Social Science & Humanities. Finland Academic Research Science Publishers
ISSN: 2945-4492 (online) | (SJIF) = 7.502 Impact factor
Volume-11| Issue-4| 2023 Published: |22-04-2023| Publishing centre of Finland FOYDALANILGAN ADABIYOTLAR: 1 Munavvarqori Abdurashidxonov. “Qizil O’zbekiston” . Toshkent. 1927 yil. Ziyo uz.com. 1 Munavvarqori Abdurashidxonov. “Qizil O’zbekiston” . Toshkent. 1927 yil. Ziyo uz.com. 1 Munavvarqori Abdurashidxonov. “Qizil O’zbekiston” . Toshkent. 1927 yil. Ziyo uz.com. 2. Qosimov B. “Milliy uyg’onish”, T “Sharq” 2004-yil, 53-bet. 3. Baxrom Irzayev. “O’zbek yoshlari va xorijiy ta’lim”, T. 2018-yil 4.Mahmudxo’ja Behbudiy . “Behbudiya kutubxonasi”,Tanlangan asarlar. Jild -
I.T.:2018-yil 14-bet 5.Sh.G’afforov. “Istibdod
davrida
Turkistondagi
ta’lim
tizimi”.-
Samarqand.2001-yil 5.Sh.G’afforov. “Istibdod
davrida
Turkistondagi
ta’lim
tizimi”.-
Samarqand.2001-yil 6. “Jadidchilik:islohat,yangilanish, mustaqillik va taraqqiyot uchun kurash”-T
Sharq 1999-yil. 6. “Jadidchilik:islohat,yangilanish, mustaqillik va taraqqiyot uchun kurash”-T
Sharq 1999-yil. 7. M.F.Zikrullayev. “Katta Qirg’in” tarixining muhim sanalari”1937-
1938yillardagi. T-“Fan” nashriyoti. 2013-yil 7. M.F.Zikrullayev. “Katta Qirg’in” tarixining muhim sanalari”1937-
1938yillardagi. T-“Fan” nashriyoti. 2013-yil 1. https://uz.m.wikipediya.org.wiki/jadidchilik 2. https://fayllar.org/jadidchilik-va-jad-adabiyoti-yangi-o’zbek-
adabiyotining-oziga.html 2. https://fayllar.org/jadidchilik-va-jad-adabiyoti-yangi-o’zbek-
adabiyotining-oziga.html 2. https://fayllar.org/jadidchilik-va-jad-adabiyoti-yangi-o’zbek-
adabiyotining-oziga.html 943
|
https://openalex.org/W3119766967
|
https://zenodo.org/records/2026985/files/article.pdf
|
English
| null |
Progress in Airships. Forms of Gas Bags
|
Scientific American
| 1,907
|
public-domain
| 3,034
|
ROMAN
FOUNDATIONS-THE
WALLS
AT
RICH·
BOROUGH, KENT, ENGLAND. In
the
absence
of
experience
in
constructing
gas
bags, those buoyant bodies have been neglected, while
the inventor struggled with the mechanism of power,
propulsion,
and guidance. The
commonplace sausage
type of gas bag supplied an
apparently easy solution
of the buoyant problem, and the fitting of some kind
of net for harnessing the bag to the suspended car
was
easier. This
has
been
especially
the
case
in
America, and most of the airships have been of that
type. ,
,
AT Richborough the foundations were thus arranged:
First, two rows of boulder stones lie on the natural
soil, which is a solid pitsand;
then a thin stratum of
chalk nodules; next a single row of boulders, and over
them another thin layer of small chalk, all these being
without cement; then boulders again, mixed with mor
tar. And so the masonry proceeds internally, with a
confused mixture of large boulders, ochre stones, sand
stone, and blocks of chalk, the whole cemented with a
mortar formed of lime, grit, large and small pebbles,
sea shells, and fragments of baked bricks. The out
side of the great northeast wall is very beautiful to
the eye, as well as magnificent. It is composed, as far
as now remains, in general, of seven great and fairly
distinct rows of stone, each of them very nearly 4 feet
thick, and each of them consisting in general of seven
courses of separate stones. The measure of the great
combined courses sometimes varies a little, some being
4 feet 3 inches, while others are only 3 feet 3 inches
in breadth, or rather in depth, which may, therefore,
perhaps, indicate an intention of forming them about
4 Roman feet in breadth or depth upon an average. These great courses of stone are separated from each
other by six smaller courses of bricks, composed each
merely of a double row of bricks that are about 1 %
inches or 1% inches in thickness, but are a f very dif
ferent breadths, from 8 inches to 1 foot;
and of very
different lengths, some being 14 inches, some 16 inches
long, and some 17% inches, a variation of dimensions
to be met with in some other Roman structures. For
in the old wall of Verulam was a brick, now worked
up in ,the wall of the abbey at St. FEBRUARY 9, 1907. 26007 causes an eddy or vortex, the violence of which rep
resents wasted impelling force. The erratic course of
the sphere is wasted travel, but is the easiest means
of relief. Hence even a flag wriggles to get past the
wind which waves it. m Santos
Dumont
are
examples. This
breaking
tend
ency is partly obviated by use of the extended pole
keel, or the triangular section frame made conspicu
ous in Santos Dumont's
airships,
extensively
copied
in this country, and provocative of "pitching" in con
nection with loss of gas. This was remedied to some
extent in the Santos Dumont airships by use of the in
ternal air bag. The lurching of gas from one end to
the other becomes a dangerous feature in connection
with fixed attachment of the suspension lines to a re
inforce built in the gas bag to replace netting, which
better readjusts the variable strains. After
these
considerations,
the
question
is
forced
upon us:
Which iight will finally remain victor, and
supplant the usual systems of illumination? Electric
ity,
illuminating
gas,
petroleum,
alcohol,
and acety
lene have entered into keen competition. Even if en
ormous strides have been made with the new metallic
flIament lamps, the efficient results of which, for the de
velopment
of electro-technics,
as
a
whole,
give
rise
to great hopes,
nevertheless it
is probable that none
of the
existing systems of illumination will soon en
tirely supplant the others. The field is too large, and
even if still more new sources of light should be dis
covered, there is room for all to exist side by side. The light of the old carbon filament lamp is certainly
dearer than in all other lamps. Hence it is possible that
in future consumers will not employ the carbon fila
ment lamp with its cheap cost of production so widely,
on account of its high working expenses. Now, if any cylinder has its ends capped with hemi
spheres, the same phenomenon occurs, modified by the
long sides of the sausage shape. The front air pressure
and the rear sucti'm oscillate the body less, while the
ail' pressure flows along the sides in waves of alternate
ly greater and less pressure rearward. The body de
sires to wriggle like the flag. PROGRESS IN AIRSHIPS.
FORMS OF GAS BAGS.*
By CARL E. MYERS. PROGRESS in airships seems directed
principally to
improvement or increase of motive power and varia
tions in the application of the propelling features. Not
much consideration
seems to be given to the proper
form of gas bag or hull, its adaptation to the environ
ment, facility of evolution, or speed, or its mainten
ance of equilibrium, or long-continued flight. The crude idea seems to be that any gas body may
be
pushed speedily through the air if the propelling
activity is sufficient. The correct idea should be to improve the form of
gas bag so as to obtain great buoyancy and efficiency
with little resistance to progress. The true principle
is to increase the propelling effect and decrease the
opposition
to
passage. This
seems
obvious,
just
as
does the fact that a sharp-pointed nail or needle may
be driven
through material
substance with less force
and less resistance than when blunted. I next devised all elongated balloon by omitting a
considerable number of segments from a sphere. With
this balloon, sharp at top and bottom, I made a per
pendicular
ascension
from
Akron,
Ohio,
September,
1879, and a fall of two miles in two minutes to a safe
landing exactly on the spot I had selected from above. The balloon ascended with its neck closed and lashed
to the concentrating ring upon which I sat, without a
basket,
my
feet
hanging below. Thus I formed the
steering rudder going up, and
the guiding prow
in
descending, and the vessel followed my extended legs
in any direction, as does the body of any bird steered
as always by its beak and extended neck. Yet a recent airship, the largest in the world, the
"Zeppelin," portrayed on the cover of the SCIENTIFIC
AMERICA=" of December 22, 1906, resembles a floating
log, with rounded ends, of 38 feet diameter, 410 feet
long,
367,000
cubic
feet
capacity,
equipped
with
70
horse-power,
driving
four propellers. It presumes
a
speed of 3 3 % miles per hour. Comparison of this aerial log with the fine lines and
clipper shape of a modern steamship shows an obvious
rliscrellancy. Tile
Parseval airship,
illustrated
in the SCIENTIFIC
A1rERlCA] of Nov,Ƥmber 10, also has a gas bag of this
sausage type, about 157 feet long and 26 feet diameter. ROMAN
FOUNDATIONS-THE
WALLS
AT
RICH·
BOROUGH, KENT, ENGLAND. Albans, which is
very nearly 2 feet in length, and there is one at Dover
near 3 feet in length. The composition of these bricks
is also as various as their dimensions. Some of them
are entirely red throughout their whole substance (like
our modern bricks), only of a deeper color;
some are
red
on
the
outside,
but
of
deep
blue
within,
the
internal substance being formed of a, different
earth
from the outside (perhaps for the sake of sparing the
better and scarcer material). And here, again, we find
a great similarity to other Roman works, for in the
walls of Chesterford, Verulam, and Silchester are ex
actly the same varied appearances. Some of the bricks
also, here at Richborough, are of a yellow color, hav
ing plainly been composed only of mud and clay taken
from the neighboring shore. And some of these latter
might
possibly have been
merely
dried by the sun;
but how the red ones should become of that color with
out the aid of fire, or how: any (except the yellow ones)
should have been dried in the sun, as has beeIY hastily
conjectured DY some antiquaries, is incomprehensible. Let them have been formed how they will, the whole
produces still a very beautiful effect to the eye. The
structure is everywhere uniformly of this sort of style,
except in some very few parts, where reparations have
plainly been made in Saxon times, and with squared
stones of a much smaller size and with herring-bone
work.-The Architect and Contract Reporter. Whether the new tungsten lamps will fulfill the far
reaching expectations placed in them, cannot, of course,
be proved
until
they have received a thorough trial
under
practical
conditions. Electric
lighting
with
glow lamps has always been a luxury. With a con
sumption of 0.5 watt per candle, electric light will be
just about as cheap as the incandescent gas light, pro
vided the price of electric energy remains what it is at
present, namely, 5 to 6 pence per kilowatt-hour. One of the earliest practical airships carrying a man
was that of C. F. Ritchel, in 1878. It was a gas bag
of about 13 feet diameter by 26 feet long, forming a
cylinder
having
nearly
flat
ends. Broad
webbing
bands
passed
over
this
cylinder
suspended
the
car
below. FEBRUARY 9, 1907. This rearward-flowing
pressure, acting as skin friction, restrains the forward
movement of the body in proportion to the surface ex
posed,
which is greater
in the
sausage than
in the
sphere, bulk for bulk. While the use of
attached suspension lines instead
of netting lessens skin friction during flight, it is an
element of weakness in securing a gas bag to its an
chorage, when exposed to wind. On the other hand, I
have had an inflated airship secured within its own
netting
exposed
for
days
without
loss
of
gas
in
a
windy "norther" blowing 20 to 30 miles per hour, an
chored with
sand bags,
its
sharp
point
toward
the
wind. In a properly-shaped gas bag the ordinary skin fric
tion is a greater cause of retarded speed than the so
called
"head
resistance," which has no actual exist
ence. Hence we should for an airship seek such forms
of body as have greatest capacity with least surface
resistance,
and
such entering and departing lines of
pressure as leave the air least disturbed, or else con
vert the disturbed waves into useful work. There is talk of a tungsten lamp having four times
the illuminating power of the carbon
filament lamp,
namely, a specific consumption of energy of about 0.8
watt per candle;
some even anticipate 0.5 watt per
candle, and the end of the incandescent gas light. It
is
still, however, a
question whether we shall soon
really possess an electric lamp so economical as only
to need 0.5 watt per candle. At present it is only the
optimists who think so, since the new tungsten lamps
are not yet out of the laboratory stage of manufacture. The makers are
not particularly willing to
publish
important results prematurely. We have already seen
that a large number of inventors are working on the
production of the tungsten lamp; it must, however, be
noted that in all countries where an examination of
patents in respect to novelty takes place, particularly
in Germany and Austria, none of the above-mentioned
patent applications except the first one by Just have up
to the
present
been
accepted. It is
yet to be seen
which of the various processes can secure effective pro
tection by patent, and it is not yet known which of
them in practice will prod lIce efficient lamps. ROMAN
FOUNDATIONS-THE
WALLS
AT
RICH·
BOROUGH, KENT, ENGLAND. A small four-bladed fan extended in front of
the car and drew it in any direction which its change
able face determined. It was an exceedingly effective
mechanism,
operated
by
hand
cranks
turned by
its
rider. M. W. QuinlHn. The
nearly flat
ends
of the
vessel prevented speedy movement. This log form is
the worst known. The
difficulty
of
producing
high-voltage
filaments
from metals with high fusing points, the necessity of
using the lamps in prescribed positions, the delicacy or
the
filament substances,
efc.-all these have almost
completely been overcome, or, at least, set aside. Coincident with this I conceived a gas kite which
avoided this obstacle, and had much greater power. This was a kite surface surmounted on its upper side
with the half of a split balloon. It looked like an up
turned
boat
having
sharp
prow
and
stern. It
was
drawn forward and -upward by a large screw-sail re
volved by hand and foot power, combined with "chain
less gear."
It mounted facing the wind, drawn up an
inclined aerial plane, raising about 50 pounds in ex
cess of its gas buoyancy. At a mile elevation "I was
monarch of all I surveyed," and could easily fall in
any direction by gravity, guiding by simply leaning to
right or left, or forward to descend, as its center of
gravity was easily
displaced. This machine was su
perior
in
accomplishment
to
anything
I
have
ever
seen. It
did
not
satisfy
my needs as
an
exhibitor,
because it could not be operated in all weather, as my
balloons
could, and it was difficult to land this gas
parachute in a wind. FEBRUARY 9, 1907. For safety, economy, carrying capacity, speed, and
easy management in air and on earth, the gas bag
should have sharp points,
and greatest bulk
amid
ships, or where the greatest weight is carried. Either
its points or its entire envelope should be continuously
distended under some pressure. Its first requisite is a
positively impervious
gas
envelope. How I have
ob
tained this I will explain later at some length. In the effort to attain such shapes I have performed
numberless
experiments, and some dangerous or dar
ing air flights,
extending
over
a quarter century of
active professional operations with hydrogen gas ves
sels, and I have tested in air, water, snow, and earth
various
forms
of
projectile
bodies,
known
and
un
known. I propose to review hereafter some of these
most useful as
object lessons in aeronautics. PROGRESS IN AIRSHIPS.
FORMS OF GAS BAGS.*
By CARL E. MYERS. "The Ville
de
Paris,"
illustrated
in
the
SCIENTIFIC
AMERICAX of November 17, is shaped forward like an
elongated egg, and aft is sausage shaped, with twin
links
of smaller
size
attached along
its
four sides. As a seas hip it would be a monstrosity, impossible to
propel at half the speed of a plain sausage. I cite the
late samples as the possible limit of absurd airship
construction of to-day,
yet
no protest against
them
has appeared. I had carried a reliable aneroid barometer, an ane
mometer, and watch. I knew how much excess buoy
ancy I had, and my ballast in small bags was weighed. Consequently, I knew the effective effort of my buoy
ant speed upward, and the equivalent of foot-pounds
effort and speed in descending. Upon nearing earth
I cast loose the neck of the balloon, which arose by
air pressure, forming a resisting surface, and I struck
the earth without harm, in a corn field. The experi
ment was successful, and not expensive. It was the
prelude to the construction of a line of variable types
of
airships
which
have
been
remarkably
successful
with
hand
and
foot
propulsion. One
such
airship
made twelve successive flights during five weeks, and
returned to the place of departure each time. Another,
provided with its own motor, one-fourth horse-power,
made a test speed of 12. miles per hour, and ran nearly
700 miles during five weeks' operation. SCIENTIFIC AMERICAN SUPPLEMENT No. 1623. SCIENTIFIC AMERICAN SUPPLEMENT No. 1623. © 1907 SCIENTIFIC AMERICAN. INC. PROTECTING PIPES
FƥOM
FREEZING. THE
means
generally
employed
to
prevent
pipes
from freezing consist in
the use
of
coatings which
protect against cold, and non-conductors of heat, such
as straw, cork, and oakum. There are, however, more
effective agents,
also
practicable
for
use in
thawing
frozen pipes. The pipes are first covered with a thin
layer
of straw,
sawdust
or
tanbark. Pieces
of
un
slaked lime as large as the fist are then packed arounrl
them, and enveloped
in another layer
of some non
conducting material, straw, oakum, or cork, and the
whole is held firmly together by means of a ",rapping
of coarse linen. The first layer is for the purpose of
protecting the pipes from the action of the fresh lime,
which would cause the metal to rust. The lime draws
moisture from the air and the materials surrounding
it. and is made warm by means of the chemical re
action. The outer covering allows only a small amount
of atmospheric air to pass through, so that the lime Practical tests show that the
resistance
of air
to
the passage of a gas body is chiefly of three sorts,
which may be termed head pressure, rear suction, and
surface
friction. A
sphere
obviously
has
the
least
surface for friction. If a sphere
is moved
through
air, there accumulates directly in front a cushion of
compressed air, most resistant here. This air relieves
its pressure by alternately dodging on either side of
the sphere,
w'hich
reciprocates
this
action
by
itself
lurching
or
pitching
in
a
zigzag
course. This
can
readily be observed by towing a small gas balloon on
a
string. Meanwhile a partial vacuum exists at the
rear, where returning air flows inward, at the expense
of
the
propelling
force. Eaph
or
these
movementR
S
I ll
d f
h
S
S Returning to the consideration of the sausage-shape
gas bag, I have observed in every case where it was
forced hard the front end or its part adjacent "caved
in" or became hollow at a certain speed, which it could
not then
exceed. Pitching from loss of gas next re
sulted. Another defect of this form of gas bag is the tend
ency to sink in the middle. of which the "Greth" air
ship, of San Francisco, and the earliest airship of M. * SpecIally prepared forthe SCIENTIFIC AllERICAN SUPPLEMENT. © 1907 SCIENTIFIC AMERICAN. INC.
|
https://openalex.org/W2904311315
|
https://europepmc.org/articles/pmc6299591?pdf=render
|
English
| null |
Expression of microRNAs in cerebrospinal fluid of dogs with central nervous system disease
|
Acta veterinaria Scandinavica
| 2,018
|
cc-by
| 3,959
|
© The Author(s) 2018. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License
(http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium,
provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license,
and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/
publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Acta Veterinaria Scandinavica Acta Veterinaria Scandinavica Marioni‑Henry et al. Acta Vet Scand (2018) 60:80
https://doi.org/10.1186/s13028-018-0434-0 Open Access Abstract In this pilot study we investigated the expression of 14 microRNAs in the cerebrospinal fluid (CSF) of dogs with neo‑
plastic, inflammatory and degenerative disorders affecting the central nervous system (CNS). CSF microRNA (miRNA)
expression profiles were compared to those from dogs with neurological signs but no evidence of structural or
inflammatory CNS disease. Seven miRNAs were easily detected in all samples: miR-10b-5p, miR-19b, miR-21-5p, miR-
30b-5p, miR-103a-3p, miR-124, and miR-128-3p. Expression of miR-10b-5p was significantly higher in the neoplastic
group compared to other groups. There was no relation between miRNA expression and either CSF nucleated cell
count or CSF protein content. Higher expression of miR-10b-5p in the neoplastic group is consistent with previous
reports in human medicine where aberrant expression of miR-10b is associated with various neoplastic diseases of
the CNS. Keywords: Canine, Cerebrospinal fluid, microRNA, Neoplasms miR-181c in the CSF of dogs with meningoencephalitis of
unknown origin (MUO) has been reported [5].h Expression of microRNAs in cerebrospinal
fluid of dogs with central nervous system
disease Katia Marioni‑Henry1* , Debiao Zaho1, Pablo Amengual‑Batle2, Nina Marie Rzechorzek1,3
and Michael Clinton1 *Correspondence: Katia.Marioni‑Henry@ed.ac.uk
1 Royal (Dick) School of Veterinary Studies and Roslin Institute, University
of Edinburgh, Easter Bush Campus, Roslin, Midlothian EH25 9RG, UK
Full list of author information is available at the end of the article Findings MicroRNAs (miRNAs) are a class of small, non-coding
RNAs of approximately 20–25 nucleotides in length that
regulate gene expression at the post-transcriptional level
[1]. MicroRNA molecules are extremely stable and miR-
NAs expressed in neurons can be found in cerebrospinal
fluid (CSF) and serum [2]. For central nervous system
(CNS) disorders, proximity to the diseased tissue and low
cellular content make CSF the preferred fluid for analyses
[1–3]. The primary aim of this pilot study was to assess the
expression of 14 miRNAs in CSF samples of dogs that
underwent a diagnostic work-up for investigation of neu-
rological signs. The secondary aim was to compare the
miRNA profiles of dogs diagnosed with structural CNS
disorders (i.e. inflammatory, neoplastic, and degenera-
tive conditions affecting brain, spinal cord and meninges;
see groups A, B and D in Table 1, which were combined
for this comparison) to those of dogs without evidence
of structural CNS disease (combined groups C and E);
this second group included dogs with epilepsy but no
evidence of a specific underlying cause (i.e. idiopathic
epilepsy group C in Table 1) and dogs with neurologi-
cal signs localised to peripheral nerves only (group E). A
tertiary aim was to compare the expression of the miR-
NAs in CSF samples of dogs in each of the five groups
listed in Table 1. The selection of the miRNAs was based
on similar studies performed in human medicine that
identified dysregulation of these 14 miRNAs in a variety In human medicine, miRNAs in CSF have been con-
sidered as potential biomarkers for the early diagnosis
and prognosis of various CNS neoplastic, inflammatory
and degenerative diseases (e.g. gliomas, glioblastomas,
metastatic brain tumours, multiple sclerosis, amyo-
trophic lateral sclerosis, Alzheimer’s disease and Parkin-
son’s disease) [1, 2, 4]. Recently, expression of miR-21 and *Correspondence: Katia.Marioni‑Henry@ed.ac.uk
1 Royal (Dick) School of Veterinary Studies and Roslin Institute, University
of Edinburgh, Easter Bush Campus, Roslin, Midlothian EH25 9RG, UK
Full list of author information is available at the end of the article Marioni‑Henry et al. Findings Acta Vet Scand (2018) 60:80 Page 2 of 6 Table 1 Clinical data for inflammatory condition group (A), neoplastic condition group (B), idiopathic epilepsy group (C),
degenerative condition group (D), and other neurological disorders not affecting the CNS group (E)
SOD1: Canine degenerative myelopathy is a progressive neurodegenerative disease associated with the c.118G > A substitution in exon 2 of the canine superoxide
dismutase 1 (SOD1) gene in German Shepherd Dogs and other affected canine breeds
FE female entire, FN female neutered, ME male entire, MN male neutered
a Diagnosis supported by histopathology
b Diagnosis supported by clinical examination, MRI findings, CSF analysis and cytology
Case
Signalment
Final diagnosis
Breed
Age (years)
Sex
A
Bernese Mountain Dog
1
FN
Steroid-responsive meningitis-arteritis
A
Medium size mixed breed
9
MN
Idiopathic polyradiculoneuritis
A
Bichon Frise
2
FN
Meningoencephalitis of unknown origin
A
Dalmatian
0.5
FE
Steroid-responsive meningitis-arteritis
B
American Bulldog cross
9
ME
Mixed glioma—braina
B
Miniature Schnauzer
7
ME
Suspected histiocytic sarcoma—braina
B
Flat Coat Retriever
8
MN
Histiocytic sarcoma—lumbar spinal corda
B
Boxer
6
FN
Glioma—brain
C
Labrador Retriever
2
FN
Idiopathic epilepsy
C
Collie cross
8
FN
Idiopathic epilepsy
C
Collie
3
MN
Idiopathic epilepsy
C
Staffordshire Bull Terrier cross
7
FN
Idiopathic epilepsy
C
Giant Schnauzer
7
FN
Idiopathic epilepsy
C
Bearded Collie
6
FN
Idiopathic epilepsy
D
German Shepherd Dog
4
MN
Canine degenerative myelopathy—SOD1 homozygous
D
German Shepherd Dog
10
FN
Canine degenerative myelopathy—SOD1 homozygousa
E
Cavalier King Charles Spaniel
6
FN
Vestibular signs associated with otitis media-internab
E
Border Collie
14
FN
Geriatric idiopathic vestibular diseaseb
E
Labrador Retriever
12
FE
Geriatric idiopathic vestibular diseaseb
E
Border Terrier
9
MN
Iliopsoas sarcomaa of two or more of the following: clinical findings, mag-
netic resonance imaging (MRI), CSF analysis, and post-
mortem examination (Table 1). of CNS neoplastic, inflammatory and degenerative dis-
eases. Teplyuk et al. Findings Relative expression was calculated as fold-change by
the ΔΔCt method with UniSp6 as a reference followed
by normalisation to the expression for sample C4 (con-
trol with idiopathic epilepsy) and to the volume of CSF
available. SD, n = 10). Expression of miR-10b-5p was also sig-
nificantly (P= 0.008) higher in the neoplastic group (B)
when compared to all other groups (Figs. 1, 2). No other
significant differences in miRNA expression between
groups were detected. However, the expression of miR-
128 approached significance with a probability level of
P = 0.074 in the idiopathic epilepsy group (C) when com-
pared to all other groups (Figs. 1, 2).i Cycle threshold (Ct) values are inverse to the quan-
tity of target nucleic acid in the sample and high Ct
values (above 35 cycles) can also be generated by deg-
radation of the probe-based fluorophore by nonspe-
cific amplification of background nucleic acids [10]. Accordingly, only Ct values < 32 were considered reli-
able and only miRNAs with Ct < 32 in the majority of
samples were included in subsequent analyses. Analy-
sis of variance (ANOVA) was used to determine if the
expression of each miRNA differed among dogs with
and without CNS structural disease and then among
the five categories of disease (Table 1). A general lin-
ear model (GLM) was used to assess the relationship
between CSF nucleated cell count and miRNA expres-
sion, and CSF protein content and miRNA expression. All statistical analyses were conducted with R version
3.2.2 [(2015–08–14) 2015, The R Foundation for Sta-
tistical Computing].i No significant correlation (P > 0.05, GLM) was detected
between CSF nucleated cell count and miRNA expres-
sion, or CSF protein content and miRNA expression. Higher expression of miR-10b-5p in the neoplastic
group is consistent with previous reports in human med-
icine; levels of both miR-10b and miR-21 were signifi-
cantly increased in the CSF of patients with glioblastoma
and brain metastasis of breast and lung cancer compared
with tumours in remission and a variety of non-neoplas-
tic conditions, while miR-10b was not detected in brain
or CSF of non-cancer human patients [6]. A recent study investigating the expression of miR-
21 and miR-181c in the CSF of dogs with MUO, found
that the expression of miRNAs in the CSF, particularly
miR-21, was correlated with the CSF nucleated cell
count [5]. Findings [6] reported increased levels of miR-
10b and miR-21 in the CSF of patients with glioblastoma
and brain metastasis of breast and lung cancer, compared
with tumors in remission and a variety of non-neoplastic
conditions; miR-21 and miR-19b were among the most
highly upregulated miRNA in primary CNS lymphoma;
miR-181c and miR-633 miR-128-3p miR-155-5p were
upregulated in multiple sclerosis, dysregulation of miR-
210 and MiR-922 miR-103a-3p miR-194-5p was reported
in degenerative conditions [7] and dysregulation of
miR146, miR155 and miR124 in epilepsy [8, 9].h CSF collected via cerebellomedullary cisternal or lum-
bar puncture was centrifuged and the supernatant was
frozen within 60 min of collection and stored at − 80 °C
until further use. All samples were visually inspected and
pink, red, or xanthochromic samples were excluded. CSF
analysis and cytology was available for 17/20 samples
(Table 2). RNA was isolated from a minimum of 250 µL and a
maximum of 500 µL of CSF. RNA extraction and cDNA
synthesis were performed using miRCURY LNA Uni-
versal RT microRNA PCR Starter Kit (Exiqon). Syn-
thetic exogenous spike-in UniSp6 RNA was added into
each sample. Real-time quantitative PCR (qRT-PCR)
analysis was performed using the ExiLENT SYBR Green
master mix (Exiqon) and the Stratagene Mx3000P The study was conducted in compliance with the guide-
lines of the Veterinary Ethical Review Committee of the
Royal (Dick) School of Veterinary Studies of the Univer-
sity of Edinburgh (Approval Number 124.17; 20 Novem-
ber 2017). All animals were examined by a board-certified
neurologist, and diagnoses were based on a combination Marioni‑Henry et al. Acta Vet Scand (2018) 60:80 Page 3 of 6 Table 2 Cerebrospinal fluid analysis (normal laboratory reference range for nucleated cells ≤ 5 cells/μL and for protein
concentration ≤ 0.25 mg/dL for cisternal samples and ≤ 0.45 mg/dL for lumbar samples)
Groups
Available data
Nucleated cell count
per μL (mean)
RBCs
Protein
mg/dL
(mean)
Inflammatory (group A)
4/4
1.1
2/4 rare
11.25
Neoplastic (group B)
2/4
4.4
Rare and moderate
numbers
122
Idiopathic epilepsy (group C)
6/6
1.1
None
18.8
Canine degenerative myelopathy (group D)
2/2
2
None
20
Neurological signs not localised to CNS or meninges
(group E)
3/4
0
None
30.6 real-time quantitative PCR system (Life Technologies). Findings This finding was not replicated in our study
using a GLM, possibly due to the low average nucleated
cell count and the small sample size of this pilot study. In our study, the CSF samples were visually inspected
prior to analysis and those presenting a reddish or xan-
thochromic discoloration were excluded to avoid con-
tamination from peripheral blood. Therefore, acute
forms of steroid responsive meningitis-arteritis (SRMA)
associated with a high nucleated cell count and xan-
thochromic CSF would have been excluded by this study
(Table 2). However, whilst highly inflammatory CSF
samples do not pose a diagnostic challenge for SRMA, Seven miRNAs were amplified in all samples: miR-
10b-5p, miR-19b, miR-21-5p, miR-103a-3p, miR-124,
and miR-128-3p. The remaining seven miRNAs were
not detected (Ct ≥ 32 in ≥ 50% of the samples): miR-146,
miR-155-5p, miR-181c, miR-194-5p, miR-210, miR-633,
and miR-922. Expression of miR-10b-5p was significantly (P= 0.028),
higher (mean 4.51 ± 0.79 SD, n = 10) in the group of
dogs with CNS disease (combined groups A, B and D)
compared to dogs without evidence of CNS structural
disease (combined groups C and E; mean 2.28 ± 0.52 Marioni‑Henry et al. Acta Vet Scand (2018) 60:80 Page 4 of 6 0
1
2
3
4
5
6
7
8
Rela ve expression
A B C D E
miR 10b-5p
*
miR 19b 0
1
2
3
4
5
6
7
8
Rela ve expression
A B C D E
miR 10b-5p
*
0
0.5
1
1.5
2
2.5
Rela ve expression
A B C D E
miR 19b
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
Rela ve expression
A B C D E
miR 21b-5p
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
Rela ve expression
A B C D E
miR 30b-5p ◂ chronic SRMA cases can present with normal or only
slightly elevated CSF protein content and mild pleocy-
tosis [11, 12]. For more than 20 years researchers have
been looking for biomarkers to improve the diagnostic
and prognostic accuracy of SRMA [11, 12]. Based on our
results, we believe that further studies investigating the
use of upregulated miRNAs as biomarkers for chronic
SRMA are warranted. For similar reasons, we decided
to include cases of acute idiopathic polyradiculoneuri-
tis (AIP) in our study. Findings In the first instance, canine AIP
is an inflammatory condition of the ventral nerve roots
and peripheral nerves analogous to Guillain–Barré syn-
drome in humans where presence of CNS inflammation
has been demonstrated by immunohistochemistry [13,
14]. Secondly, like chronic SRMA, canine AIP is diag-
nostically challenging since it presents with abnormal
CSF analysis characterized by nonspecific elevated pro-
tein content and normal nucleated cell count [13]. Our
primary objective was therefore to investigate the pos-
sibility of detecting upregulated miRNAs in the CSF of
AIP cases as potentially useful diagnostic biomarkers.h 0
0.5
1
1.5
2
2.5
Rela ve expression
A B C D E
miR 21b-5p 0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
Rela ve expression
A B C D E p
y
g
The major limitations of this pilot study were the low
sample size and the lack of a true control group of dogs
without neurological signs; this to comply with UK legis-
lation that requires that any medical procedure involving
client-owned dogs must be performed for the direct ben-
efit of the patient. Studies performed on human patients
face similar limitations in regard to control groups, how-
ever, CSF is still considered the ideal source of miRNA
to investigate brain disorders in human patients, due to a
profile identical to that of the brain tissue and a less inva-
sive and more accessible procedure compared to a brain
biopsy [7]. 0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
Rela ve expression
A B C D E
miR 30b-5p Fig. 1 Relative expression of microRNA-10b-5p (miR-10b-5p),
microRNA-19b (miR-19b), microRNA-21b-5p (miR-21b-5p),
microRNA-30b-5p (miR-30b-5p) in cerebrospinal fluid (CSF) of dogs
with inflammatory conditions (group A), neoplastic conditions (group
B), idiopathic epilepsy (group C), degenerative conditions (group D),
and other neurological disorders not affecting the CNS (group E). Error
bars represent the standard error. *Statistically significant difference
between groups (*P = 0.008) Authors’ contributions KMH and PAB developed the research concept; NMR and PAB were respon‑
sible for sample collection and storage. KMH, MC and DZ developed the
methods. DZ was a major contributor in quantitative real-time PCR analysis. KMH, DZ and MC analysed data and KMH performed the statistical analyses. KMH prepared the manuscript with contributions from MC, NMR and PAB. All
authors read and approved the final manuscript. Author details 1 Royal (Dick) School of Veterinary Studies and Roslin Institute, University
of Edinburgh, Easter Bush Campus, Roslin, Midlothian EH25 9RG, UK. 2 School
of Veterinary Medicine, College of Veterinary, Medical and Life Sciences, Uni‑
versity of Glasgow, Bearsden Road, Glasgow G61 1QH, UK. 3 Centre for Clinical
Brain Sciences, University of Edinburgh, Chancellor’s Building, 49 Little France
Crescent, Edinburgh EH16 4SB, UK. 0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1.8
2
Rela ve expression
A B C D E
iR 128 3 Abbreviations AIP: acute idiopathic polyradiculoneuritis; ANOVA: analysis of variance; cDNA:
complementary DNA; CNS: central nervous system; CSF: cerebrospinal fluid;
Ct: cycle threshold; GLM: general linear model; miRNA or miR: microRNAs;
MRI: magnetic resonance imaging; MUO: meningoencephalitis of unknown
origin; qRT-PCR: real-time quantitative PCR; RBC: red blood cells; SRMA: steroid-
responsive meningitis-arteritis; SOD1: superoxide dismutase 1. Acknowledgements
Th
h
ld lik The authors would like to acknowledge Mrs Emma Jeffery for helping with
sample collection and Dr. Ted Henry for help with statistical analysis. Funding This work was supported by the Fiona and Ian Russell Seed Corn Fund for
Companion Animal Research Grant (Grant Number G31335). Preliminary
results were presented at the 30th Annual Symposium of the European Col‑
lege of Veterinary Neurology, Helsinki, Finland, 21–23 September 2017. The Roslin Institute receives Institute Strategic Grant funding from the
BBSRC (M.C. & D.Z.). Fig. 2 Relative expression of microRNA-103a-3p (miR-103a-3p),
microRNA-124 (miR-124), microRNA-128-3p (miR-128-3p) in
cerebrospinal fluid (CSF) of dogs with inflammatory conditions
(group A), neoplastic conditions (group B), idiopathic epilepsy
(group C), degenerative conditions (group D), and other neurological
disorders not affecting the CNS (group E). Error bars represent the
standard error. *Statistically significant difference between groups
(*P = 0.008) Fig. 2 Relative expression of microRNA-103a-3p (miR-103a-3p),
microRNA-124 (miR-124), microRNA-128-3p (miR-128-3p) in
cerebrospinal fluid (CSF) of dogs with inflammatory conditions
(group A), neoplastic conditions (group B), idiopathic epilepsy
(group C), degenerative conditions (group D), and other neurological
disorders not affecting the CNS (group E). Error bars represent the
standard error. *Statistically significant difference between groups
(*P = 0.008) Fig. 2 Relative expression of microRNA-103a-3p (miR-103a-3p),
microRNA-124 (miR-124), microRNA-128-3p (miR-128-3p) in
cerebrospinal fluid (CSF) of dogs with inflammatory conditions
(group A), neoplastic conditions (group B), idiopathic epilepsy
(group C), degenerative conditions (group D), and other neurological
disorders not affecting the CNS (group E). Error bars represent the
standard error. *Statistically significant difference between groups
(*P = 0.008) N.M.R. was funded by a Wellcome Trust Integrated Training Fellowship for
Veterinarians (096409/Z/11/Z). Conclusionsh The expression of 14 miRNAs in the CSF of dogs with
neurological disorders with and without structural CNS
abnormalities showed that 7 miRNAs were consistently
expressed in all samples. We also found a significantly
increased expression of miR-10b-5p in the neoplastic
group compared to other groups. Marioni‑Henry et al. Acta Vet Scand (2018) 60:80 Page 5 of 6 0
0.5
1
1.5
2
2.5
Rela ve expression
A B C D E
miR 103a-3p
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1.8
2
Rela ve expression
A B C D E
miR 124
0
0.5
1
1.5
2
2.5
Rela ve expression
A B C D E
miR 128-3p
Fig. 2 Relative expression of microRNA-103a-3p (miR-103a-3p),
microRNA-124 (miR-124), microRNA-128-3p (miR-128-3p) in
cerebrospinal fluid (CSF) of dogs with inflammatory conditions
(group A), neoplastic conditions (group B), idiopathic epilepsy
(group C), degenerative conditions (group D), and other neurological
disorders not affecting the CNS (group E). Error bars represent the
standard error. *Statistically significant difference between groups
(*P = 0.008) 0
0.5
1
1.5
2
2.5
Rela ve expression
A B C D E
miR 103a-3p
miR 124 Competing interests None of the authors of this paper has a financial or personal relationship with
other people or organisations that could inappropriately influence or bias the
content of the paper. Availability of data and materials The datasets used and/or analyzed during the current study are available from
the corresponding author on reasonable request. Publisher’s Note Springer Nature remains neutral with regard to jurisdictional claims in pub‑
lished maps and institutional affiliations. Springer Nature remains neutral with regard to jurisdictional claims in pub‑
lished maps and institutional affiliations. Received: 15 August 2018 Accepted: 13 December 2018 If our findings are replicated in a larger cohort, CSF
miRNA expression analysis may prove to be a useful,
minimally invasive technique to support the pre-mortem
diagnosis of CNS disorders in dogs. Consent for publication 0
0.5
1
1.5
2
2.5
Rela ve expression
A B C D E Ethics approval and consent to participate The study was conducted in compliance with the guidelines of the Veterinary
Ethical Review Committee of the Royal (Dick) School of Veterinary Studies of
the University of Edinburgh approval number 124.17; 20 November 2017. References . Burgos K, Malenica I, Metpally R, Courtright A, Rakela B, Beach T, et al. Profiles of extracellular miRNA in cerebrospinal fluid and serum from
patients with Alzheimer’s and Parkinson’s diseases correlate with disease
status and features of pathology. PLoS ONE. 2014;9:e94839. https://doi. org/10.1371/JOURNAL.PONE.0094839. Page 6 of 6 Marioni‑Henry et al. Acta Vet Scand (2018) 60:80 Marioni‑Henry et al. Acta Vet Scand (2018) 60:80 2. Shalaby T, Grotzer AM. Tumor-associated CSF microRNAs for the predic‑
tion and evaluation of CNS malignancies. Int J Mol Sci. 2015;16:29103–19.f 9. Brennan GP, Dey D, Chen Y, Patterson KP, Magnetta EJ, Hall AM, et al. Dual and opposing roles of microRNA-124 in epilepsy are mediated
through inflammatory and NRSF-dependent gene networks. Cell Rep. 2016;14:2402–12. https://doi.org/10.1016/j.celrep.2016.02.042. 3. Burgos KL, Javaherian A, Bomprezzi R, Ghaffari L, Rhodes S, Courtright
A, et al. Identification of extracellular miRNA in human cerebrospinal
fluid by next-generation sequencing. RNA. 2013;19:712–22. https://doi. org/10.1261/rna.036863.112. 10. Caraguel CGB, Stryhn H, Gagné N, Dohoo IR, Hammell KL. Selection of a
cutoff value for real-time polymerase chain reaction results to fit a diag‑
nostic purpose: analytical and epidemiologic approaches. J Vet Diagn
Invest. 2011;23:2–15. https://doi.org/10.1177/104063871102300102. 10. Caraguel CGB, Stryhn H, Gagné N, Dohoo IR, Hammell KL. Selection of a
cutoff value for real-time polymerase chain reaction results to fit a diag‑
nostic purpose: analytical and epidemiologic approaches. J Vet Diagn
Invest. 2011;23:2–15. https://doi.org/10.1177/104063871102300102. 4. Benigni M, Ricci C, Jones AR, Giannini F, Al-Chalabi A, Battistini S. Identi‑
fication of miRNAs as potential biomarkers in cerebrospinal fluid from
amyotrophic lateral sclerosis patients. NeuroMol Med. 2016;18:551–60. https://doi.org/10.1007/s12017-016-8396-8. 11. Bathen-Noethen A, Carlson R, Menzel D, Mischke R, Tipold A. Con‑
centrations of acute-phase proteins in dogs with steroid responsive
meningitis-arteritis. J Vet Intern Med. 2008;22:1149–56. https://doi. org/10.1111/j.1939-1676.2008.0164.x. 11. Bathen-Noethen A, Carlson R, Menzel D, Mischke R, Tipold A. Con‑
centrations of acute-phase proteins in dogs with steroid responsive
meningitis-arteritis. J Vet Intern Med. 2008;22:1149–56. https://doi. org/10.1111/j.1939-1676.2008.0164.x. 5. Gaitero L, Russell SJ, Monteith GLJ. Expression of microRNAs miR-21
and miR-181c in cerebrospinal fluid and serum in canine meningoen‑
cephalomyelitis of unknown origin. Vet J. 2016;216:122–4. https://doi. org/10.1016/J.TVJL.2016.07.014. 12. Tipold A, Jaggy A. Steroid responsive meningitis-arteritis in dogs: long-
term study of 32 cases. J Small Anim Pract. 1994;35:311–6. https://doi. org/10.1111/j.1748-5827.1994.tb03293.x. 6. Teplyuk NM, Mollenhauer B, Gabriely G, Giese A, Kim E, Smolsky M, et al. MicroRNAs in cerebrospinal fluid identify glioblastoma and metastatic
brain cancers and reflect disease activity. Neuro-oncology. References 2012;14:689–
700. https://doi.org/10.1093/neuonc/nos074. 13. Cuddon PA. Acquired canine peripheral neuropathies. Vet Clin
Small Anim Pract. 2002;32:207–49. https://doi.org/10.1016/S0195
-5616(03)00086-X. 13. Cuddon PA. Acquired canine peripheral neuropathies. Vet Clin
Small Anim Pract. 2002;32:207–49. https://doi.org/10.1016/S0195
-5616(03)00086-X. 7. Stoicea N, Du A, Lakis DC, Tipton C, Arias-Morales CE, Bergese SD. The
miRNA journey from theory to practice as a CNS biomarker. Front Genet. 2016;7:11. https://doi.org/10.3389/fgene.2016.00011. 14. Müller HD, Beckmann A, Schröder JM. Inflammatory infiltrates in the
spinal cord of patients with Guillain–Barré syndrome. Acta Neuropathol. 2003;106:509–17. https://doi.org/10.1007/s00401-003-0768-0. 14. Müller HD, Beckmann A, Schröder JM. Inflammatory infiltrates in the
spinal cord of patients with Guillain–Barré syndrome. Acta Neuropathol. 2003;106:509–17. https://doi.org/10.1007/s00401-003-0768-0. 8. Srivastava A, Dixit AB, Banerjee J, Tripathi M, Sarat Chandra P. Role of
inflammation and its miRNA based regulation in epilepsy: implications
for therapy. Clin Chim Acta. 2016;452:1–9. https://doi.org/10.1016/j. cca.2015.10.023. •
fast, convenient online submission
•
thorough peer review by experienced researchers in your field
•
rapid publication on acceptance
•
support for research data, including large and complex data types
•
gold Open Access which fosters wider collaboration and increased citations
maximum visibility for your research: over 100M website views per year
•
At BMC, research is always in progress. Learn more biomedcentral.com/submissions
Ready to submit your research ? Choose BMC and benefit from: •
fast, convenient online submission
•
thorough peer review by experienced researchers in your field
•
rapid publication on acceptance
•
support for research data, including large and complex data types
•
gold Open Access which fosters wider collaboration and increased citations
maximum visibility for your research: over 100M website views per year
•
At BMC, research is always in progress. Learn more biomedcentral.com/submissions
Ready to submit your research ? Choose BMC and benefit from:
|
https://openalex.org/W3097491155
|
https://www.frontiersin.org/articles/10.3389/fmars.2020.587686/pdf
|
English
| null |
Population Genetic Structure and Gene Expression Plasticity of the Deep-Sea Vent and Seep Squat Lobster Shinkaia crosnieri
|
Frontiers in marine science
| 2,020
|
cc-by
| 13,251
|
Yao Xiao1†, Ting Xu1,2†, Jin Sun1, Yan Wang1, Wai Chuen Wong1, Yick Hang Kwan1,
Chong Chen3, Jian-Wen Qiu2* and Pei-Yuan Qian1* 1 Department of Ocean Science and Hong Kong Branch of the Southern Marine Science and Engineering Guangdong
Laboratory (Guangzhou), The Hong Kong University of Science and Technology, Hong Kong, China, 2 Department of Biology,
Hong Kong Baptist University, Hong Kong, China, 3 X-STAR, Japan Agency for Marine-Earth Science and Technology
(JAMSTEC), Yokosuka, Japan ORIGINAL RESEARCH
published: 02 November 2020
doi: 10.3389/fmars.2020.587686 ORIGINAL RESEARCH
published: 02 November 2020
doi: 10.3389/fmars.2020.587686 Overall, exploring the population Reviewed by:
Yong-Jin Won,
Ewha Womans University,
South Korea
Yun-wei Dong,
Xiamen University, China
*Correspondence:
Jian-Wen Qiu
qiujw@hkbu.edu.hk
Pei-Yuan Qian
boqianpy@ust.hk
†These authors have contributed
equally to this work Reviewed by:
Yong-Jin Won,
Ewha Womans University,
South Korea
Yun-wei Dong,
Xiamen University, China *Correspondence:
Jian-Wen Qiu
qiujw@hkbu.edu.hk
Pei-Yuan Qian
boqianpy@ust.hk
†These authors have contributed
equally to this work Specialty section:
This article was submitted to
Marine Molecular Biology
and Ecology,
a section of the journal
Frontiers in Marine Science
Received: 27 July 2020
Accepted: 22 September 2020
Published: 02 November 2020 Specialty section:
This article was submitted to
Marine Molecular Biology
and Ecology,
a section of the journal
Frontiers in Marine Science Received: 27 July 2020
Accepted: 22 September 2020
Published: 02 November 2020 Edited by:
Yong Wang,
Institute of Deep-Sea Science
and Engineering (CAS), China Edited by:
Yong Wang,
Institute of Deep-Sea Science
and Engineering (CAS), China Edited by:
Yong Wang,
Institute of Deep-Sea Science
and Engineering (CAS), China Edited by:
Yong Wang,
Institute of Deep-Sea Science
and Engineering (CAS), China Shinkaia crosnieri (Decapoda: Munidopsidae) is a squat lobster that dominates both
deep-sea hydrothermal vent and methane seep communities in the Western Pacific. Previous studies comparing S. crosnieri living in these two types of habitats have
suffered from methodological and/or sample size limits. Here, using transcriptome-wide
single nucleotide polymorphisms (SNPs) markers from 44 individuals of S. crosnieri,
we reveal the extent of genetic connectivity between a methane seep population
in the South China Sea and a hydrothermal vent population in the Okinawa
Trough, as well as their signatures of local adaptation. Analysis of differentially
expressed genes (DEGs) between these two populations and population-specific
genes (PSGs) revealed that a large number of unigenes, such as cytochrome
P450 (CYP), glutathione S-transferase (GST) and peroxiredoxin 6 (Prdx6) related to
oxidoreductase, and sulfur dioxygenase (ETHE1) and chondroitin 4-sulfotransferase
11 (CHST11) related to sulfur metabolism, showed opposite expression patterns in
these two populations. Data subsampling in this study revealed that at least five
individuals of S. crosnieri per site are required to generate reliable results from the
differential gene expression analysis. Population genetic analyses based on 32,452
SNPs revealed clear genetic differentiation between these two populations with an
FST value of 0.07 (p < 0.0005), and physical oceanographic modeling of the ocean
currents in middle and deep layers also suggests a weak connection between
these two sites. Analysis of outlier SNPs revealed 345 unigenes potentially under
positive selection, such as sarcosine oxidase/L-pipecolate oxidase (PIPOX), alanine-
glyoxylate transaminase/serine-glyoxylate transaminase/serine-pyruvate transaminase
(AGXT), and Cu-Zn superoxide dismutase (SOD1). Among the differentially expressed
genes and genes with amino acid substitutions between the two sites are those related
to oxidation resistance and xenobiotic detoxification, indicating local adaptation to
the specific environmental conditions of each site. INTRODUCTION or three mitochondrial genes (COI, cytochrome b gene (Cytb),
and 16S rRNA) (Shen et al., 2016) have revealed clear genetic
differentiation between a methane seep population in the SCS
and hydrothermal vent populations in the OT. Nevertheless, due
to limited genomic coverage, a single to a few gene markers can
hardly reflect the population divergence at the genome level and
do not allow for the effective discovery of signatures of natural
selection. Genome-wide single-nucleotide polymorphism (SNP)
markers have the potential to resolve these issues (Xu et al., 2012). Recently, Cheng et al. (2020) obtained 12,963 genome-wide SNPs
using restriction site associated DNA sequencing (RAD-Seq). By
using these markers, they also revealed clear genetic divergence
between a seep population of S. crosnieri in the SCS and a
vent population in the OT. Nevertheless, RAD-Seq can only
capture a reduced representation of the genome, and numerous
SNP markers are located in non-coding regions (Pegadaraju
et al., 2013; Houston et al., 2014). Indeed, Cheng et al. (2020)
identified 54 outlier SNPs potentially under positive selection,
but only five were in the protein-coding regions. More recently,
Cheng et al. (2019) compared the transcriptomes of S. crosnieri
collected from the same seep and vent populations, but with
only three individuals from each population. They detected 545
differentially expressed genes (DEGs) and 82 protein-coding
genes (PCGs) that have potentially undergone positive selection. However, the small sample size may have limited the statistical
power for the detection of DEGs and PCGs. Deep-sea hydrothermal vents, often distributed along active mid-
ocean ridges and back-arc spreading centers, are well known
for discharging sulfur-rich geofluids into the water column
(Corliss et al., 1979; German et al., 2000). Methane seeps, usually
found along the continental margins and in trenches, typically
release methane-rich geofluids from the seabed more slowly (Van
Dover et al., 2002). These two types of ecosystems share some
similar features, such as lack of light to support photosynthesis,
high pressure, and high concentration of chemically reduced
compounds as well as heavy metals (Levin, 2005; German et al.,
2011). Nevertheless, a great number of macrobenthos flourish
in these ‘extreme’ environments, forming high-biomass hotspots
powered by chemosynthesis in the deep ocean. Over 700 species
of macrobenthos have been recorded in the global hydrothermal
vents and more than 600 in the methane seeps (German et al.,
2011). INTRODUCTION However, only a small fraction of them inhabit both
hydrothermal vents and methane seeps (Watanabe et al., 2010;
Vrijenhoek, 2013), indicating that thriving in both types of
environments requires specific adaptation and gene expression
(Watanabe et al., 2010). The Western Pacific is an ideal region for a comparative
study of vent and seep populations due to their close proximity
and the presence of 20% of all the recorded macrobenthos
in both types of habitats (Watanabe et al., 2010). Kiel (2016)
reported the Western Pacific as having the highest number of
active, sedimentary back-arc vents in the world, which might
provide a vital biogeographic link between vent and seep animals. Previous population genetic studies of deep-sea macrobenthos in
the Western Pacific have mainly focused on macrobenthos with
a planktotrophic larval stage, such as bathymodioline mussels
(Kyuno et al., 2009; Miyazaki et al., 2010; Xu et al., 2017,
2018) and the deep-sea limpet Shinkailepas myojinensis (Yahagi
et al., 2017). In this study, we aim to understand the population
connectivity and local environmental adaptation of the squat
lobster Shinkaia crosnieri (Decapoda: Munidopsidae), whose
larvae are lecithotrophic and likely have a limited dispersal ability
(Nakajima et al., 2018). Shinkaia crosnieri was initially discovered
on the Edison Seamount vent field in the Bismarck Archipelago
(Baba and Williams, 1998), and was later found in methane seeps
in the South China Sea (SCS) and hydrothermal vents in the
Okinawa Trough (OT), with a known bathymetric range between
700 and 2,200 m (Chan et al., 2000; Watanabe et al., 2010;
Miyazaki et al., 2017a). The distribution pattern of S. crosnieri
in terms of both habitat types and life-history trait makes it a
suitable model for studies of population divergence and local
adaptation under different environmental conditions. In the present study, we sequenced the transcriptomes of a
total of 44 S. crosnieri individuals, including 20 from a methane
seep in the SCS and 24 from a hydrothermal vent in the OT. We identified DEGs between the two populations, population-
specific genes (PSGs), transcriptome-wide SNP markers, and
outlier SNP markers. We also conducted subsampling analyses
to determine the number of individuals required to generate
representative data for a meaningful population comparison. Our
results not only provide new insights into the local adaptation
and population genetics of S. crosnieri inhabiting both types
of habitats, but also demonstrate how different techniques and
sample sizes may affect these results. Citation: Xiao Y, Xu T, Sun J, Wang Y,
Wong WC, Kwan YH, Chen C,
Qiu J-W and Qian P-Y (2020)
Population Genetic Structure
and Gene Expression Plasticity of the
Deep-Sea Vent and Seep Squat
Lobster Shinkaia crosnieri. Front. Mar. Sci. 7:587686. doi: 10.3389/fmars.2020.587686 November 2020 | Volume 7 | Article 587686 Frontiers in Marine Science | www.frontiersin.org 1 Xiao et al. Population Connectivity and Deep-Sea Adaptation structure of S. crosnieri using transcriptome-wide SNP markers resulted in an improved
understanding of its molecular adaptation and expression plasticity in vent and
seep ecosystems. structure of S. crosnieri using transcriptome-wide SNP markers resulted in an improved
understanding of its molecular adaptation and expression plasticity in vent and
seep ecosystems. Keywords: deep sea, gene expression, population connectivity, RNA-seq, single-nucleotide polymorphism Frontiers in Marine Science | www.frontiersin.org Transcriptome Assembly, Completeness
Assessment, and Functional Annotation Quantitative real-time reverse transcription-PCR (qRT-PCR)
was performed to validate the expression levels of selected genes. Primers for qRT-PCR were designed utilizing the online NCBI
Primer-BLAST tool5 with sequences given in Supplementary
Table 1. Total RNA of two individuals (SCS_23 and OT_3)
were extracted with the TRIzol kit (Thermo Fisher Scientific,
United States) and reverse transcription from RNA to single-
stranded cDNA was conducted using the High Capacity
cDNA Reverse Transcription Kit (Thermo Fisher Scientific,
United States). The SYBR Green PCR Master Mix (Thermo
Fisher Scientific, United States) was utilized with synthesized
cDNA to conduct qPCR reaction on a LightCycler 480
Instrument II (Roche, Switzerland). The relative fold change in
the expression of selected genes was calculated by the 2−11Ct
method using actin as an internal standard gene. The correlation
coefficient (r2) between expression of qRT-PCR and that of RNA-
Seq was analyzed in Excel. Assessment, and Functional Annotation
The quality of raw sequencing data was controlled using
FASTQC2 and filtered using TRIMMOMATIC v.0.36 (Bolger
et al., 2014). The parameters were leading: 10; trailing: 10; sliding
window: 4:15; minlen: 25, and adapters: ILLUMINACLIP:
TruSeq3-PE.fa:2:30:10. The
resultant
clean
reads
of
the
four tissues from OT_1, testis from OT_2 and abdominal
muscle from SCS_3 (with maximum sequence data size
of SCS individuals) were de novo assembled using Trinity
v.2.8.3 (Grabherr et al., 2011) under default settings except
the min_contig_length parameter was set to 300 and the
min_kmer_cov was set to 2. The longest isoform of each
gene cluster was selected using a custom Python script as a
unigene. CD-HIT-EST (Fu et al., 2012) was used to remove
redundant unigenes based on a similarity threshold of 95%. TransDecoder v.5.5.0 (Haas et al., 2013) was then applied
to predict candidate open reading frames (ORFs) with the
single_best_only option. BUSCO v.3.0 (Simão et al., 2015)
was utilized to evaluate the completeness of the final assembly
based on the metazoa_odb9 database, and the Perl script
assemblathon_stats.pl (Bradnam et al., 2013) was run to evaluate
the final assembly. The transcriptome from each individual was
also assembled separately using the same Trinity parameters and
CD-HIT was applied thereafter. Gene Ontology enrichment analysis of DEGs was performed
with Fisher’s Exact Test in OmicsBox v1.3.116. KEGG pathway
enrichment analysis was performed via Enrichr (Chen et al.,
2013). In both enrichment analyses the significant enrichment
level was set to 0.05. Sample Collection A total of 44 S. crosnieri individuals were used in this study. Among them, 10 were collected by the manned submersible
Jiaolong from Jiaolong Ridge (also known as the F site), a
methane seep (22◦06.921′ N, 119◦17.131′ E; 1,122 m deep) in the
SCS in June 2013. Another set of 10 individuals were collected
using the remotely operated vehicle (ROV) ROPOS on board
the research vessel (R/V) Tan Kah Kee from the same site in Previous studies of S. crosnieri using the mitochondrial
cytochrome c oxidase submit I (COI) gene (Yang et al., 2016) November 2020 | Volume 7 | Article 587686 Frontiers in Marine Science | www.frontiersin.org 2 Population Connectivity and Deep-Sea Adaptation Xiao et al. April 2018. In addition, 24 individuals were collected using the
ROV KAIKO (with vehicle Mk-IV) from the Sakai hydrothermal
field (27◦31.4749′ N, 126◦59.021′ E; 1,550 m deep) (Nakamura
et al., 2015) in the OT during the Japan Agency for Marine-Earth
Science and Technology (JAMSTEC) R/V Kairei cruise KR15-17
in November 2015. Samples were frozen at −80◦C upon recovery
or fixed with RNAlater, and then shipped to the laboratory and
stored at −80◦C until use. 10−5 and the online annotation KAAS-KEGG server3 applying
the single-directional best hit method was used to search
for the Kyoto Encyclopedia of Genes and Genomes (KEGG)
annotation. Differentially Expressed Genes (DEGs)
and Population-Specific Genes (PSGs)
Analyses Kallisto (Bray et al., 2016) was used to quantify abundances of
the assembled unigenes with a bootstrap value of 100. The gene
expression level in transcripts per kilobase million (TPM) was
further normalized with the TMM method in edgeR (Robinson
et al., 2010). The correlation between individuals and normalized
TPM was determined by principal component analysis (PCA)
implemented in the R package DESeq2 (Love et al., 2014). Genes
without expression were removed, and the average TPM of each
contig was calculated individually within each sampling site. The
Differential expression analysis was conducted via the web portal
RNA-seq 2G (Zhang et al., 2017) using the DESeq2 method with
a minimal read count of 10. Unigenes with an absolute value of
fold change greater than 2 and a false discovery rate (FDR) less
than 0.05 were considered as DEGs. Partial DEGs were visualized
by Morpheus4. 2https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ 1www.novogene.com 1www.novogene.com
2https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ RNA Extraction and Sequencing The gill, hepatopancreas, ovary, and abdominal muscle from
one vent individual (OT_1), testis, and abdominal muscle from
another vent individual (OT_2) and the abdominal muscles of
the other 42 individuals (from SCS_1 to 20 and OT_3 to 24) were
dissected for total RNA extraction using the TRIzol kit (Thermo
Fisher Scientific, United States) according to the manufacturer’s
protocol. The quantity and quality of RNA were examined using
1% agarose gel electrophoresis and NanoDrop 2000 (Thermo
Fisher Scientific, United States), respectively. RNA libraries were
constructed individually and sequenced on an Illumina HiSeq
2500 platform (PE150) by Novogene Bioinformatics Technology
Co., Ltd., Beijing, China1. 1www.novogene.com
2https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ November 2020 | Volume 7 | Article 587686 Frontiers in Marine Science | www.frontiersin.org Transcriptome Assembly, Completeness
Assessment, and Functional Annotation The transcriptome data for individuals collected from the
two sampling sites were compared, adopting the same Kalisto
and RNA-seq 2G parameters as mentioned above. The groups
of orthologous genes of each individual transcriptome were
detected using Proteinortho v.6.0b (Lechner et al., 2011). PSGs
were defined as the unigenes shared among at least 90% of
the individuals from one site but among less than 10% of the
individuals from the other site (Parra et al., 1998; Belcaid et al., All obtained unigenes were searched against the NCBI
non-redundant (NR) protein database using BLASTp v.2.7.1
(Altschul et al., 1990) with an E-value of 10−5, a word size
of 3, a minimum alignment of 20, and max hsps of 20, and
the resultant .xml file was fed into OmicsBox (Götz et al.,
2008) to search for the Gene Ontology (GO) function. The
UniProt database was scanned using BLASTp with an E-value of November 2020 | Volume 7 | Article 587686 Frontiers in Marine Science | www.frontiersin.org 3 Population Connectivity and Deep-Sea Adaptation Xiao et al. Detection and Annotation of Outlier
SNPs 2019). PSGs were annotated according to the NCBI NR and
KEGG databases. The KEGG pathway of PSGs was reconstructed
using KEGG Mapper7. Two methods were adopted to detect outlier SNPs. First, a
coalescent method implemented in Arlequin v.3.5 (Excoffier and
Lischer, 2010) was used for outlier detection by running 20,000
simulations with 100 simulated demes per group, and SNPs with
a p-value < 0.05 were considered as the Arlequin outliers. Second,
a multivariate analysis was implemented in the R package pcadapt
(Luu et al., 2017) for outlier identification, and SNPs with an
adjusted p-value < 0.1 from the Benjamini–Hochberg procedure
were considered as the pcadapt outliers. Outliers detected by
both methods were retained and used for GO enrichment and
KEGG enrichment analyses using the same approaches described
in the Section “Differentially Expressed Genes and Population-
Specific Genes”. Statistical
power
of
different
sample
sizes
used
for
the identification of DEGs was tested by a subsampling
approach. In brief, a total of 3, 5, 10, and 15 individuals
were randomly selected from each habitat to identify DEGs
using
the
methods
detailed
in
this
section. Afterward,
comparison
was
conducted
to
determine
an
appropriate
sample size for the reliable DEGs. In addition, DEGs obtained
in our study were also compared with those reported in
Cheng et al. (2019). RESULTS Statistical power of different sample sizes selected to identify
SNPs was tested via a subsampling approach as well. In brief,
a total of 3, 5, and 10 individuals were randomly selected from
each habitat to identify SNPs using the methods described above. Afterward, the SNP numbers were compared to evaluate the
impact of sample sizes on SNP detection. 7https://www.genome.jp/kegg/tool/map_pathway.html
8http://picard.sourceforge.net Identification of Transcriptome-Wide
SNP Markers Raw reads of each individual were trimmed using Trimmomatic
v.0.36 (see section “Transcriptome Assembly, Completeness
Assessment, and Functional Annotation” for the settings). Filtered reads of each individual were mapped to the assembled
transcriptome using Bowtie2 (Langmead and Salzberg, 2012)
with a maximum fragment length of 1,200 (–maxins 1200), a
seed of 20 (−D 20), re-seed reads twice (−R 2), a mismatch
number of 1 (−N 1), a seed length of 18 (−L 18), and an
interval between seeds of S,1,0.50. The generated .sam files were
converted to the .bam format and sorted by SAMtools v.1.8
(Li et al., 2009). PCR duplicates were removed using Picard
MarkDuplicates8. Then, the mpileup function implemented
in SAMtools was applied to generate .bcf files with the
following settings: 10 for minimum mapping quality for an
alignment (−q 10) and 20 for minimum base quality (−Q
20). SNPs were identified using the call function implemented
in BCFtools v.1.9 (Li, 2011) with the default settings along
with the options −mv and −Ov. The obtained .vcf file
for each individual was then sorted using VCFtools v.0.1.13
(Danecek et al., 2011) and the .vcf files of all the individuals
were combined into a single file using the merge function
in BCFtools v.1.9. 9https://www.hycom.org/ Population Genetic Analyses Based on
the Entire and the Outlier SNP Dataset
and Physical Oceanographic Modeling Genetic differentiation was evaluated using the pairwise Fst
statistic via Arlequin v.3.5.2.2 with the default settings based on
both the entire and the outlier SNP datasets. Population structure
was examined using the maximum-likelihood estimation method
implemented in ADMIXTURE v1.3 (Alexander et al., 2009) and
PCA implemented in the R package SNPRelate (Zheng et al.,
2012) using single SNP per locus of both the entire and the outlier
SNP datasets. The .ped, .map, and .bed format files (Alexander
et al., 2015) required by the ADMIXTURE analysis were
transformed based on the combined .vcf file of all the individuals
using PLINK v.2.00a2LM (Purcell et al., 2007) with the –allow-
extra-chr option. The number of ancestral populations (i.e., the
K value) in ADMIXTURE was predefined from 1 to 3. When
K was set to 2, the divergence was between sites. Population
structure was visualized using the function bar plot in R based
on the best K value. The relative migration pattern of all samples
was estimated via divMigrate in the R package diveRsity (Keenan
et al., 2013) using the DJost statistic as a measure of genetic
distance and a bootstrap value of 1,000. To validate population
genetics results against physical oceanographic model of larval
drift, we inferred the annual-mean lateral ocean currents from
the HYCOM + NCODA Global 1/12◦Reanalysis (experiment
sequence: 53.X) data9, which has assimilated multiple sources of
available observational records of the ocean. Filtering of SNPs was carried out using VCFtools v.0.1.13 with
the following criteria: (1) retaining only bi-allelic sites (–min-
alleles 2 –max-alleles 2); (2) with a minor allele frequency (MAF)
greater than or equal to 0.02 (–maf 0.02); (3) excluding sites with
indels (–remove-indels); (4) excluding sites not conforming to
Hardy–Weinberg equilibrium with a p-value threshold of 0.01;
(5) retaining SNPs successfully genotyped in at least 50% of all
individuals (–max-missing 0.5); and (6) retaining SNPs with a
sequencing depth greater than 3 and less than 120 (–minDP 3
and –maxDP 120). Transcriptome Assembly, Completeness,
and Annotation A total of 3.93 Gb of paired-end clean reads of abdominal
muscle from one SCS individual (SCS_3) and 10.96 Gb of paired-
end clean reads of five tissues (gill, ovary, abdominal muscle,
hepatopancreas, and testis) from two OT individuals (OT_1 and
OT_2) (Supplementary Table 2) were obtained after quality November 2020 | Volume 7 | Article 587686 Frontiers in Marine Science | www.frontiersin.org 4 Population Connectivity and Deep-Sea Adaptation Xiao et al. FIGURE 1 | (A) Principal component analysis (PCA) based on gene expression matric of all individuals shows complete separation between individuals from the
South China Sea (SCS) and the Okinawa Trough (OT). Red dots indicate samples collected from the OT, and yellow and blue dots indicate samples collected from
SCS in 2013 and 2018, respectively. (B) Relationship between FDR and fold change. Red and green dots represent up-regulated genes in the SCS and the OT,
respectively. Yellow dots indicate non-DEGs. Vertical dotted lines: log2 (Fold change) = 1 and –1; horizontal dotted line: –log10 (FDR) = 0.05. FIGURE 1 | (A) Principal component analysis (PCA) based on gene expression matric of all individuals shows complete separation between individuals from the
South China Sea (SCS) and the Okinawa Trough (OT). Red dots indicate samples collected from the OT, and yellow and blue dots indicate samples collected from
SCS in 2013 and 2018, respectively. (B) Relationship between FDR and fold change. Red and green dots represent up-regulated genes in the SCS and the OT,
respectively. Yellow dots indicate non-DEGs. Vertical dotted lines: log2 (Fold change) = 1 and –1; horizontal dotted line: –log10 (FDR) = 0.05. metabolism’ (ko00640), ‘spliceosome’ (ko03040), and ‘focal
adhesion’ (ko04510). Meanwhile, 13 OT-PSGs were mapped to 26
KEGG pathways, such as ‘drug metabolism – cytochrome P450’
(ko00982), ‘peroxisome’ (ko04146), and ‘glycosaminoglycan
biosynthesis - chondroitin sulfate/dermatan sulfate’ (ko00532). control and were used for transcriptome reference assembly. The detailed sequencing statistics of the abdominal muscle from
these individuals are summarized in Supplementary Table 3. De
novo assembly and data filtering resulted in 29,273 unigenes. The
assembled transcriptome had a contig N50 of 2,690 bp and a
mean contig size of 1,615 bp. The contig length ranged from 301
to 29,765 bp. BUSCO analysis revealed that the transcriptome
contained 89.7% complete plus 3.1% fragmented conserved
metazoan genes. Transcriptome Assembly, Completeness,
and Annotation Results of PCA based on the unigene expression matric
revealed that all the SCS seep individuals were separated from all
the OT vent individuals along the first eigenvector (Figure 1A). Further analyses showed that a total of 4,854 (16.6%) unigenes
were differentially expressed between S. crosnieri from the SCS
and the OT. Among them, 2,597 (8.87%) unigenes showed
higher expression levels in the SCS than in the OT, whereas
2,257 (7.71%) had higher expression levels in the OT than in
the SCS. Relationship between the FDR and fold change for
all DEGs is illustrated in Figure 1B. Expression heatmap and
hierarchical clustering analysis of 29 DEGs involved in oxidative
activity, metabolism of xenobiotics by cytochrome P450, sulfur
metabolism, and methane metabolism are presented in Figure 2
and their mean expression levels in the SCS and OT as well
as functional annotations are given in Supplementary Table 6. Furthermore, qRT-PCR and RNA-Seq data are highly correlated
(R2 = 0.97) (Supplementary Figure 2), indicating the robustness
of our DEG analyses. g
A total of 19,743 (67.44%) unigenes had at least one significant
hit in the NR, UniProt, KEGG, or GO database. In total, 7,044
(24.06%) unigenes were annotated according to the GO database
and assigned to three major GO categories according to GO level
three in OmicsBox: biological process (BP, 707, 2.42%), molecular
function (MF, 1,394, 4.76%), and cellular component (CC, 1,589,
5.43%) (Supplementary Figure 1A). Specifically, in the category
of BP, MF, and CC, ‘organic substance metabolic process’ (GO:
0071704), ‘organic cyclic compound binding’ (GO: 0097159),
and ‘organelle’ (GO: 0043226) were the top most abundant
subcategories, respectively. Furthermore, 8,707 (29.7%) unigenes
were mapped to 295 KEGG pathways, with ‘thermogenesis,’
‘endocytosis,’ and ‘RNA transport’ being the top three most
abundant pathways (Supplementary Figure 1B). There were 40 and 32 subcategories enriched in the SCS
seep population and the OT vent population, respectively, by
GO enrichment analysis (Supplementary Figure 3A). The
top three enriched categories in the seep population were
‘translation’ (GO: 0006412), ‘structural constituent of ribosome’
(GO: 0003735), and ‘polymeric cytoskeletal fiber’ (GO: 0099513);
and those in the vent population were ‘adenyl ribonucleotide
binding’ (GO: 0032559), ‘motor activity’ (GO: 0003774), and
‘myosin complex’ (GO: 0016459). Additionally, more (40
vs. 32) subcategories were enriched in the seep population
than in the vent population. For instance, ‘oxidoreductase PSGs and DEGs Between the SCS and
OT Populations De novo transcriptome assembly for each individual produced
an average of 38,707 transcripts with statistical details presented
in Supplementary Table 4. Specifically, a total of 61 (0.16%)
and 51 (0.13%) SCS and OT PSGs were identified in our
study and their KEGG and NR annotations are presented
in Supplementary Table 5. Among them, 10 SCS-PSGs
were mapped to 18 KEGG pathways, such as ‘propanoate November 2020 | Volume 7 | Article 587686 Frontiers in Marine Science | www.frontiersin.org 5 Population Connectivity and Deep-Sea Adaptation Xiao et al. FIGURE 2 | Heatmap and hierarchical clustering analysis of 29 DEGs between the SCS and the OT. The color code represents the gene expression level based on
the Log-transformed values. Red color indicates higher expression, whereas blue color indicates lower expression. FIGURE 2 | Heatmap and hierarchical clustering analysis of 29 DEGs between the SCS and the OT. The color code represents the gene expression level based on
the Log-transformed values. Red color indicates higher expression, whereas blue color indicates lower expression. FIGURE 2 | Heatmap and hierarchical clustering analysis of 29 DEGs between the SCS and the OT. The color code represents the gene expression level based on
the Log-transformed values. Red color indicates higher expression, whereas blue color indicates lower expression. FIGURE 2 | Heatmap and hierarchical clustering analysis of 29 DEGs between the SCS and the OT. The color code represents the gene expression level based on
the Log-transformed values. Red color indicates higher expression, whereas blue color indicates lower expression. FIGURE 2 | Heatmap and hierarchical clustering analysis of 29 DEGs between the SCS and the OT. The color code represents the gene expression level based on
the Log-transformed values. Red color indicates higher expression, whereas blue color indicates lower expression. EGG enrichment analysis of all DEGs between S. crosnieri individuals from the SCS and those from the OT. TABLE 1 | KEGG enrichment analysis of all DEGs between S. crosnieri individuals from the SCS and those from the OT. PSGs and DEGs Between the SCS and
OT Populations Term
Overlap*
p-value
DEGs from SCS individuals
Glycosaminoglycan biosynthesis
8/53
1.05E-04
Pentose and glucuronate interconversions
5/34
2.40E-03
RNA transport
10/165
1.80E-02
Mannose type O-glycan biosynthesis
3/23
2.55E-02
Spliceosome
8/134
3.50E-02
Ubiquinone and other terpenoid-quinone biosynthesis
2/11
3.64E-02
DEGs from OT individuals
Vitamin digestion and absorption
3/24
1.43E-02
Mineral absorption
4/44
1.45E-02
Axon guidance
9/180
1.61E-02
Amino sugar and nucleotide sugar metabolism
4/49
2.09E-02
Bacterial invasion of epithelial cells
5/74
2.15E-02
ECM-receptor interaction
5/83
3.33E-02
Starch and sucrose metabolism
3/33
3.34E-02
ErbB signaling pathway
5/84
3.48E-02
Mitophagy
4/63
4.65E-02
*Significant terms overlap with the input list. DEGs from OT individuals *Significant terms overlap with the input list. vent population including ‘axon guidance’ (ko04360), ‘amino
sugar and nucleotide sugar metabolism’ (ko00520), and ‘bacterial
invasion of epithelial cells’ (ko05100) (Table 1). activity, oxidizing metal ions, NAD or NADP as acceptor’ (GO:
0016723) was one of the representative subcategories in the
seep population, which included metalloreductase (STEAP3). procollagen-lysine,2-oxoglutarate
5-dioxygenase
(PLOD),
tryptophan 2,3-dioxygenase (TDO2), 4-hydroxyphenylpyruvate
dioxygenase
(HPPD),
cysteine
dioxygenase
(CDO1),
and
prolyl 4-hydroxylase (P4HA) belonged to the ‘dioxygenase
activity’ (GO: 0051213) subcategory that was enriched in
the vent population. A total of 692 unigenes upregulated
in the seep population had KEGG annotations, while 589
unigenes upregulated in the vent population were annotated. The
results
of
KEGG
pathway-enriched
analysis
showed
six significantly enriched pathways in the seep population
including
‘glycosaminoglycan
biosynthesis
–
chondroitin
sulfate/dermatan sulfate’ (ko00532), ‘pentose and glucuronate
interconversions’ (ko00040), and ‘RNA transport’ (ko03013),
while eight significantly enriched pathways were detected in the activity, oxidizing metal ions, NAD or NADP as acceptor’ (GO:
0016723) was one of the representative subcategories in the
seep population, which included metalloreductase (STEAP3). procollagen-lysine,2-oxoglutarate
5-dioxygenase
(PLOD),
tryptophan 2,3-dioxygenase (TDO2), 4-hydroxyphenylpyruvate
dioxygenase
(HPPD),
cysteine
dioxygenase
(CDO1),
and
prolyl 4-hydroxylase (P4HA) belonged to the ‘dioxygenase
activity’ (GO: 0051213) subcategory that was enriched in
the vent population. A total of 692 unigenes upregulated
in the seep population had KEGG annotations, while 589
unigenes upregulated in the vent population were annotated. The
results
of
KEGG
pathway-enriched
analysis
showed
six significantly enriched pathways in the seep population
including
‘glycosaminoglycan
biosynthesis
–
chondroitin
sulfate/dermatan sulfate’ (ko00532), ‘pentose and glucuronate
interconversions’ (ko00040), and ‘RNA transport’ (ko03013),
while eight significantly enriched pathways were detected in the Frontiers in Marine Science | www.frontiersin.org *Significant terms overlap with the input list. SNP Identification and Outlier SNP
Characterization After genotyping and stringent data filtering (Table 2), a
total of 32,452 transcriptome-wide SNPs located in 8,667
unigenes were obtained and subjected to downstream analyses. Outlier screening tests resulted in 1,065 Arlequin outliers and
1,307 pcadapt outliers, with 504 outliers identified by both
approaches (Figure 3A). These 504 outlier SNPs (Figure 3B)
were located in 345 unigenes and GO enrichment analysis of
these unigenes resulted in the discovery of 27 enriched categories
(Supplementary Figure 3B). Among them, the top category
for all, as well as molecular function, was ‘ATP binding’ (GO: November 2020 | Volume 7 | Article 587686 Frontiers in Marine Science | www.frontiersin.org 6 Population Connectivity and Deep-Sea Adaptation Xiao et al. TABLE 2 | Number of filtering SNP candidates retained after each step of filtering. Category
Count
Bi-allelic
1,059,632
Overall minor allele frequency (MAF ≥0.02)
800,814
Without an indel
800,814
Hardy–Weinberg equilibrium (p ≥0.01)
683,175
Genotyped in 50% of individuals
36,705
Coverage depth ≥3 and ≤120
32,452 protein 1 (CREB1), succinate dehydrogenase cytochrome b560
subunit (SDHC), succinate dehydrogenase (SDHD), NAD-
dependent protein deacetylase Sirt6 (SIRT6), and succinate
dehydrogenase [ubiquinone] iron-sulfur subunit (SDHB). Genes
from environment-related pathways such as sulfur dioxygenase
(ETHE1) in the ‘sulfur metabolism’ (ko00920) pathway and
dihydrodiol dehydrogenase (DDH) in the ‘metabolism of
xenobiotics by cytochrome P450’ (ko00980) pathway were
identified in the outlier SNP dataset and the results of amino
acid substitutions of 27 outlier SNPs with annotation are given
in Table 5. Overall, 74 unigenes were found in both the outlier
SNP dataset and the DEG dataset (42 in the SCS and 32 in the
OT) (Supplementary Table 7), such as ETHE1 and DDH in
the SCS population, suggesting a link between mutation and
expressional change. 0070403), and the second and third categories of MF were
‘calcium ion binding’ (GO: 0005509) and ‘zinc ion binding’ (GO:
0008270), respectively. The category of ‘calcium ion binding’
includes myosin light chain 6 and 12 (MYL6 and MYL12),
chloride intracellular channel protein 2 (CLIC2), Plastin-2
(PLSL) and serine/threonine-protein phosphatase 2A regulatory
subunit B” subunit alpha (PPP2R3A); the category of ‘zinc ion
binding’ includes NAD-dependent protein deacylase 2 and 5
(SIRT2 and SIRT5), glycine hydroxymethyltransferase (GlyA)
and chitin deacetylase 1 (CDA1). The annotations and partial
functions of the nine metal ion binding related genes using the
published literature are presented in Table 3. Population Structure Based on the Entire
and the Outlier SNP Dataset Pairwise Fst calculated based on the entire SNP dataset was 0.07
(p < 0.0005), and pairwise Fst value estimated based on the outlier
SNP dataset was 0.43 (p < 0.0005), much higher than that based
on the entire SNP dataset as expected. ADMIXTURE analyses
and PCA based on both all the (Figures 4A,B) and just the
outlier SNPs (Figures 4C,D) clearly revealed two genetic groups
of S. crosnieri, with one formed by all the individuals from the
SCS and the other formed by all the individuals from the OT. Nevertheless, eight individuals in the OT showed a signature
of admixture of two genetic groups (Figure 4A), indicating a
stronger gene flow from the SCS to the OT. A similar result
was also obtained via the divMigrate analysis where a higher
migration rate was detected in the SCS to OT direction (1.00
for SCS to OT vs. 0.21 for OT to SCS). Figure 5A shows the
lateral velocity vectors at 500 m depth laid upon the bathymetry
in our study region. The Kuroshio Current is visible at this level KEGG enrichment analysis revealed that 180 of the 345
outlier-containing unigenes were significantly enriched in 15
pathways (Table 4). Specifically, a number of pathways may be
related to the adaptation of S. crosnieri to local environments,
although it is not possible to pinpoint the specific environmental
condition shaping the adaptation. For example, three unigenes
were enriched in the ‘peroxisome’ (ko04146) pathway: sarcosine
oxidase/L-pipecolate
oxidase
(PIPOX),
alanine-glyoxylate
transaminase/serine-glyoxylate
transaminase/serine-pyruvate
transaminase
(AGXT),
and
Cu-Zn
superoxide
dismutase
(SOD1); and five unigenes were enriched in the ‘thermogenesis’
(ko04714) pathway: cyclic AMP-responsive element-binding A
B
FIGURE 3 | (A) Locus-specific Fst is plotted against observed heterozygosity (Heterozygosity) with black circles indicating neutral SNPs, red circles indicating outlier
SNPs only identified by Arlequin, yellow circles indicating outlier SNPs only identified by PCAdapt and blue circles indicating overlapped outlier SNPs identified by two
methods. (B) A total of 504 outlier SNPs is plotted based on observed heterozygosity and locus-specific Fst value with seven genes highlighted with different colors. B A FIGURE 3 | (A) Locus-specific Fst is plotted against observed heterozygosity (Heterozygosity) with black circles indicating neutral SNPs, red circles indicating outlier
SNPs only identified by Arlequin, yellow circles indicating outlier SNPs only identified by PCAdapt and blue circles indicating overlapped outlier SNPs identified by two
methods. Subsampling Analyses and Comparison
With Previous Studies Although the samples we used were different from those used in
Cheng et al. (2019), some DEGs identified in our study showed
the same expression patterns as in Cheng et al. (2019), including
cystathionine gamma-lyase (CSE) and heat shock protein 22
(HSP) (Table 6), indicating they might reflect true differences
between the two populations. Nevertheless, subsampling of our
data clearly revealed a requirement of a larger sample size to
capture the DEGs of the two populations (Figure 6A). From our
data, the total number of DEGs increased sharply from 2,142
when using data from three individuals to 4,353 when using data
from five individuals. But the number of DEGs did not increase
further when data from even more individuals were included in
the analysis. from RAD-Seq. Another advantage is that SNPs detected from
transcriptome are all located in protein coding sequences
while markers from RAD-Seq were sometimes out of protein-
coding regions which is degermed by restriction enzymes
(Cheng et al., 2020). The SNP markers detected using the subsampling strategy
showed a basically consistent trend when 3, 5, and 10 individuals
were chosen from each habitat (Figure 6B), and all three
subsampling groups under transcriptome sequencing obtained
more SNPs than the number of loci obtained under RAD
sequencing. Moreover, more outlier SNP sequences were detected
from transcriptome-wide SNP dataset compared with the results Frontiers in Marine Science | www.frontiersin.org Population Structure Based on the Entire
and the Outlier SNP Dataset (B) A total of 504 outlier SNPs is plotted based on observed heterozygosity and locus-specific Fst value with seven genes highlighted with different colors. November 2020 | Volume 7 | Article 587686 Frontiers in Marine Science | www.frontiersin.org 7 Population Connectivity and Deep-Sea Adaptation Xiao et al. TABLE 3 | Nine enriched outlier-associated unigenes related with metal ion binding. GO term
Gene name
Annotation
Partial function
References
Calcium ion binding
MYL6
Myosin light chain 6
MYL6 belongs to MLC1, calcium binding proteins. Leclerc et al., 2016
MYL12
Myosin regulatory light chain 12
MYL12 regulates activity of the myosin ATPase, inhibits
interactions between myosin and actin. Calcium binding to
myosin activates the myosin ATPase and provokes muscle
contraction. Kendrick-Jones et al.,
1976
PPP2R3A
Serine/threonine-protein
phosphatase 2A regulatory
subunit B′′ subunit alpha
PPP2R3A catalyzes the removal of phosphate groups from
serine and/or threonine residues with the requirement of
manganese ions. Swingle and Honkanen,
2019
PLSL
Plastin-2
PLSL binds for Ca2+ originally, also a candidate for binding
U ions with a low pH of 5.2; involved in the formation of
cross-linked actin filaments. Bucher et al., 2016
CLIC2
Chloride intracellular channel
protein 2
CLIC2 inhibits cardiac ryanodine receptor (RyR) calcium
release channels, regulates calcium release from
intracellular stores in the heart and skeletal muscles. Cromer et al., 2007
Zinc ion binding
SIRT2
NAD-dependent deacetylase
sirtuin 2
SIRT may be related to minerals deficiency; correlated with
Zn2+ binding; SIRTs in skeletal muscle regeneration and
survival under catabolic stress (esp. 2 and 5); SIRT2 can
extend lifespan; influence metabolism, circadian rhythms,
and potentially life span through their function as protein
deacetylase. Ha et al., 2019
SIRT5
NAD-dependent deacetylase
sirtuin 5
GlyA
Glycine
hydroxymethyltransferase
GlyA catalyzes the reversible cleavage of serine to form
glycine and monocarbonic groups
Rubin-Blum et al., 2017
CDA1
Deacetylation reaction for
insoluble chitin 1
Deacetylation reaction for insoluble chitin; convert chitin into
water-soluble chitosan; metal ions such as Sr2+, Mg2+,
Na+, Ca2+, and K+ simulate the activity rate of CDA and
further inhibited by Co2+, Ba2+
Trimukhe and Varma,
2008; Chai et al., 2020 TABLE 3 | Nine enriched outlier-associated unigenes related with metal ion binding. Population Structure Based on the Entire
and the Outlier SNP Dataset GlyA catalyzes the reversible cleavage of serine to form
glycine and monocarbonic groups
Rubin-Blum et al., 2017
Deacetylation reaction for insoluble chitin; convert chitin into
water-soluble chitosan; metal ions such as Sr2+, Mg2+,
Na+, Ca2+, and K+ simulate the activity rate of CDA and
further inhibited by Co2+, Ba2+
Trimukhe and Varma,
2008; Chai et al., 2020 TABLE 4 | KEGG enrichment analysis of outlier SNPs. TABLE 4 | KEGG enrichment analysis of outlier SNPs. Term
Overlap*
p-value
Alanine, aspartate, and glutamate metabolism
4/35
2.05E-04
Glycine, serine, and threonine metabolism
4/40
3.47E-04
Mismatch repair
3/23
9.11E-04
Citrate cycle (TCA cycle)
3/30
2.00E-03
Valine, leucine, and isoleucine degradation
3/48
7.63E-03
Axon guidance
5/181
1.85E-02
Lysosome
4/123
2.02E-02
Bacterial invasion of epithelial cells
3/74
2.44E-02
RNA degradation
3/79
2.89E-02
Peroxisome
3/83
3.28E-02
ErbB signaling pathway
3/85
3.48E-02
Thermogenesis
5/231
4.57E-02
*Significant terms overlap with the input list. and turns anticyclonically at the Luzon strait, allowing for water
exchange between the Pacific Ocean and the SCS. At 1,000 m
depth, however, much less connectivity is revealed between the
Pacific and the South China Sea SCS (Figure 5B). DISCUSSION Function
Outlier SNP ID
ALT
AA Change
Iron incorporation into
metallo-sulfur cluster
TRINITY_DN23438_c2_g1_i3_1730
AAG
R/K
Lysosome
TRINITY_DN893_c2_g2_i2_1077
GTT
T/V
TRINITY_DN5263_c0_g1_i7_1262
GTC
I/V
TRINITY_DN1916_c0_g2_i2_2889
CCC
–
TRINITY_DN8085_c0_g1_i3_150
GGG
E/G
TRINITY_DN8085_c0_g1_i3_2302
CAC
–
TRINITY_DN3521_c0_g1_i14_2372
CCC
–
TRINITY_DN3521_c0_g1_i14_2392
GCG
V/A
TRINITY_DN6522_c0_g1_i2_1819
TTC
–
Endocytosis
TRINITY_DN2898_c0_g1_i3_1076
CAC
–
TRINITY_DN47762_c0_g1_i1_2457
GCG
–
TRINITY_DN5968_c0_g1_i23_1932
GCA
V/A
TRINITY_DN5968_c0_g1_i23_2234
TGT
R/C
Phagosome
TRINITY_DN5194_c3_g1_i1_115
GCC
T/A
TRINITY_DN229_c1_g1_i3_3160
TTC
–
TRINITY_DN229_c1_g1_i3_4608
GTC
L/V
TRINITY_DN1849_c0_g1_i1_1404
ATG
L/M
Peroxisome
TRINITY_DN3834_c0_g1_i2_1142
CCA
S/P
TRINITY_DN2705_c1_g1_i5_943
AGC
R/S
TRINITY_DN12145_c0_g1_i3_479
GGC
D/G
Methane metabolism
TRINITY_DN31417_c0_g2_i1_152
CGG
W/R
TRINITY_DN2705_c1_g1_i5_943
AGC
R/S
TRINITY_DN664_c1_g1_i1_829
CCG
S/P
Sulfur metabolism
TRINITY_DN595_c1_g1_i9_1123
GTG
–
C-type lectin receptor
signaling pathway
TRINITY_DN474_c0_g1_i5_967
GAA
I/Z
TRINITY_DN474_c0_g1_i5_968
ALT, alternate codon and marked with boldness; AA, amino acid; “–”, synonymous substitution. TABLE 5 | Functional classification and amino acid substitution of the 27 annotated outlier SNPs. provided the basis for investigating the population differentiation
and environment-specific local adaptation of S. crosnieri from a
methane seep and a hydrothermal vent in the Western Pacific. Luzon Strait. The Luzon Strait is the only deep channel that
connects the semi-enclosed SCS with the deeper Pacific Ocean
for water transportation via a sandwiched vertical structure
(Tian et al., 2009). Seawater flows into the SCS from the
Pacific Ocean in both the upper (<700 m) and the deep
(>1,500 m) layers and exits the SCS through the Luzon Strait
in the intermediate layer (700–1,500 m) (Tian et al., 2006; Yuan
et al., 2009). The eastward spread is stronger in both winter
and spring (You et al., 2005). Previously, by using genome-
wide SNP markers, Xu et al. (2018) revealed a limited gene
flow of the deep-sea mussel Gigantidas platifrons (previously
known as Bathymodiolus platifrons) between the SCS and the
OT. However, G. platifrons and S. crosnieri have different life-
history traits. Bathymodioline mussels produce planktotrophic
larvae that take almost a year to develop, and their dispersal DISCUSSION By using the RNA-Seq technique, a total of 112 PSGs, 4,854
DEGs, and 32,452 SNPs were detected in this study, which November 2020 | Volume 7 | Article 587686 8 Population Connectivity and Deep-Sea Adaptation Xiao et al. TABLE 5 | Functional classification and amino acid substitution of the 27 annotated outlier SNPs. Function
Outlier SNP ID
ALT
AA Change
Annotation
Iron incorporation into
metallo-sulfur cluster
TRINITY_DN23438_c2_g1_i3_1730
AAG
R/K
iscS, NFS1; cysteine desulfurase [EC:2.8.1.7]
Lysosome
TRINITY_DN893_c2_g2_i2_1077
GTT
T/V
CTSB; cathepsin B [EC:3.4.22.1]
TRINITY_DN5263_c0_g1_i7_1262
GTC
I/V
SLC17A5; MFS transporter, ACS family, solute carrier family
17 (sodium-dependent inorganic phosphate cotransporter),
member 5
TRINITY_DN1916_c0_g2_i2_2889
CCC
–
SCARB2, LIMP2, CD36L2; lysosome membrane protein 2
TRINITY_DN8085_c0_g1_i3_150
GGG
E/G
NPC1; Niemann-Pick C1 protein
TRINITY_DN8085_c0_g1_i3_2302
CAC
–
NPC1; Niemann-Pick C1 protein
TRINITY_DN3521_c0_g1_i14_2372
CCC
–
BTS, CLN3; battenin
TRINITY_DN3521_c0_g1_i14_2392
GCG
V/A
BTS, CLN3; battenin
TRINITY_DN6522_c0_g1_i2_1819
TTC
–
AP1S1_2; AP-1 complex subunit sigma 1/2
Endocytosis
TRINITY_DN2898_c0_g1_i3_1076
CAC
–
ARPC3; actin related protein 2/3 complex, subunit 3
TRINITY_DN47762_c0_g1_i1_2457
GCG
–
VPS36, EAP45; ESCRT-II complex subunit VPS36
TRINITY_DN5968_c0_g1_i23_1932
GCA
V/A
VTA1, LIP5; vacuolar protein sorting-associated protein
VTA1
TRINITY_DN5968_c0_g1_i23_2234
TGT
R/C
VTA1, LIP5; vacuolar protein sorting-associated protein
VTA1
Phagosome
TRINITY_DN5194_c3_g1_i1_115
GCC
T/A
ATPeV1A, ATP6A; V-type H+-transporting ATPase subunit
A [EC:3.6.3.14]
TRINITY_DN229_c1_g1_i3_3160
TTC
–
ITGB1, CD29; integrin beta 1
TRINITY_DN229_c1_g1_i3_4608
GTC
L/V
ITGB1, CD29; integrin beta 1
TRINITY_DN1849_c0_g1_i1_1404
ATG
L/M
CANX; calnexin
Peroxisome
TRINITY_DN3834_c0_g1_i2_1142
CCA
S/P
PIPOX; sarcosine oxidase/L-pipecolate oxidase [EC:1.5.3.1
1.5.3.7]
TRINITY_DN2705_c1_g1_i5_943
AGC
R/S
AGXT; alanine-glyoxylate transaminase/serine-glyoxylate
transaminase/serine-pyruvate transaminase [EC:2.6.1.44
2.6.1.45 2.6.1.51]
TRINITY_DN12145_c0_g1_i3_479
GGC
D/G
SOD1; superoxide dismutase, Cu-Zn family [EC:1.15.1.1]
Methane metabolism
TRINITY_DN31417_c0_g2_i1_152
CGG
W/R
glyA, SHMT; glycine hydroxymethyltransferase [EC:2.1.2.1]
TRINITY_DN2705_c1_g1_i5_943
AGC
R/S
AGXT; alanine-glyoxylate transaminase/serine-glyoxylate
transaminase/serine-pyruvate transaminase [EC:2.6.1.44
2.6.1.45 2.6.1.51]
TRINITY_DN664_c1_g1_i1_829
CCG
S/P
FBP, fbp; fructose-1,6-bisphosphatase I [EC:3.1.3.11]
Sulfur metabolism
TRINITY_DN595_c1_g1_i9_1123
GTG
–
ETHE1; sulfur dioxygenase [EC:1.13.11.18]
C-type lectin receptor
signaling pathway
TRINITY_DN474_c0_g1_i5_967
GAA
I/Z
PPP3C, CNA; serine/threonine-protein phosphatase 2B
catalytic subunit [EC:3.1.3.16]
TRINITY_DN474_c0_g1_i5_968
PPP3C, CNA; serine/threonine-protein phosphatase 2B
catalytic subunit [EC:3.1.3.16]
ALT, alternate codon and marked with boldness; AA, amino acid; “–”, synonymous substitution. TABLE 5 | Functional classification and amino acid substitution of the 27 annotated outlier SNPs. Population Genetic Structure of
Shinkaia crosnieri Between SCS and OT Previous population genetic studies (Shen et al., 2016; Yang et al.,
2016; Cheng et al., 2020) using different molecular markers have
consistently shown that the seep population of S. crosnieri in the
SCS and the vent population in the OT form two distinct genetic
groups. By using 32,452 transcriptome-wide SNPs markers, our
study also uncovered the same pattern of population structure. One of the abiotic factors that may have contributed to such
population subdivision is the physical barrier formed by the November 2020 | Volume 7 | Article 587686 Frontiers in Marine Science | www.frontiersin.org 9 Population Connectivity and Deep-Sea Adaptation Xiao et al. A
C
D
B
FIGURE 4 | Population structure analyses based on ADMIXUTRE (A,C) and PCA (B,D) using entire SNP for (A,B) and the outlier SNP dataset for (C,D), showing the
population genetic structure of S. crosnieri (K for optimal genetic group number). A B C C D FIGURE 4 | Population structure analyses based on ADMIXUTRE (A,C) and PCA (B,D) using entire SNP for (A,B) and the outlier SNP dataset for (C,D), showing the
population genetic structure of S. crosnieri (K for optimal genetic group number). FIGURE 5 | Annual-mean lateral velocity vectors, denoted by black arrows, at (A) 500 m depth and (B) 1,000 m depth laid upon the bathymetry (color contours;
unit: km) in our analysis region. Sampling sites of the SCS and the OT are indicated by the red hexagrams. Datasets are based on the HYCOM + NCODA Global
1/12◦Reanalysis (experiment sequence: 53.X). FIGURE 5 | Annual-mean lateral velocity vectors, denoted by black arrows, at (A) 500 m depth and (B) 1,000 m depth laid upon the bathymetry (color contours;
unit: km) in our analysis region. Sampling sites of the SCS and the OT are indicated by the red hexagrams. Datasets are based on the HYCOM + NCODA Global
1/12◦Reanalysis (experiment sequence: 53.X). ability is largely influenced by the upper or surface currents
(Arellano and Young, 2009). Consequently, the limited larval
exchange of G. platifrons between the two sides of the Luzon
Strait is suggested to be achieved by the looping path of the
Kuroshio Current, which flows into the SCS via the middle part and exits via the northern part of the Luzon Strait (Nan et al.,
2011), as well as by the North Pacific Intermediate Water (NPIW;
Xu et al., 2018). In contrast, S. Local Adaptation of Shinkaia crosnieri Results of DEGs, PSGs, and outlier SNP detection revealed that
several functional groups such as oxidoreductase activity, sulfur
metabolism and metal ion binding may be involved in the
local adaptation and expression plasticity of S. crosnieri under
the varying environmental conditions of methane seeps and
hydrothermal vents. Apart from cellular organelles, the mixed-function oxygenase
(MFO) system also contributes to xenobiotic detoxification
in all organisms (Ramos and Garcia, 2007). The first phase
of MFO is mainly controlled by biotransformation enzymes
like CYP and glutathione S-transferase (GST) that introduce
a functional group to transform and metabolize toxicants. Superoxide dismutase (SOD), catalase (CAT), and glutathione
peroxidase (GPx) catalyze subsequent reactions (second phase)
and generate more polar products which are more easily
excreted by the organisms (Lee, 1981). CYP and GST were
upregulated
unigenes
in
the
seep
population,
while
the
alternative hydroperoxide of GPx, the peroxiredoxin 6 (PRDX
6) that also uses glutathione (GSH) as an electron donor
(Deponte, 2013), was upregulated in the seep population. Cu-Zn superoxide dismutase (SOD1), a major antioxidant
defense enzyme in charge of the detoxification of superoxide
radicals and the generation of H2O2, was identified as an
outlier SNP. The different expression levels of MFO system-
related enzymes may suggest varying toxicant accumulation
patterns between the two studied populations. Besides SOD1,
sarcosine oxidase/L-pipecolate oxidase (PIPOX) and alanine-
glyoxylate transaminase/serine-glyoxylate transaminase/serine-
pyruvate transaminase (AGXT) were two other unigenes
enriched under the ‘peroxisome’ pathway from the outlier
SNP dataset. PIPOX catalyzes the oxidation of sarcosine and
l-pipecolate with the production of H2O2 and it was also
identified as a PSG in the vent population. AGXT takes part
in glyoxylate detoxification and prevents the generation of
toxic oxalate from glyoxylate. All three unigenes underwent
amino substitutions. Differential gene expression and allele
frequency changes of all these oxidoreductase-related genes may
suggest different extents of local adaptation to the extreme
environmental conditions. Mitochondria are a target of pollutant-induced toxicity, as
reflected by the expressional changes of mitochondria-related
proteins as well as oxidative enzymes in response to exposure
to toxicants (Gobe and Crane, 2010). Iron plays a role in many
cellular processes as well as the generation of harmful reactive
oxygen species (ROS). Extracellular iron is taken up by cells and
transported to mitochondria which form a major center of iron
utilization and accumulation. Population Genetic Structure of
Shinkaia crosnieri Between SCS and OT crosnieri produces large (2 mm)
oil-rich eggs and equally oil-rich lecithotrophic larvae (Miyake
et al., 2010; Nakajima et al., 2018), and its dispersal is expected November 2020 | Volume 7 | Article 587686 Frontiers in Marine Science | www.frontiersin.org 10 Population Connectivity and Deep-Sea Adaptation Xiao et al. in our outlier SNP dataset, and sulfur dioxygenase (ETHE1),
a homodimeric Fe-containing sulfur dioxygenase (SDO), was
a DEG in the seep population as well as an outlier SNP with
synonymous substitution. Natural gas hydrate on the seafloor
of the Formosa Ridge (also known as the Jiaolong Ridge) in
the SCS contained significant amounts of H2S (Zhang et al.,
2017). These results indicate activation of mitochondrial sulfur
detoxification in the seep population to cope with the local
physicochemical condition. One PSG in the vent population,
chondroitin 4-sulfotransferase 11 (CHST11), belongs to the
family of sulfotransferases that catalyze the conversion of sulfate
to chondroitin 4′-sulfate, a sulfated polysaccharide usually found
in the matrix of animal cells (Suzuki and Strominger, 1960;
Helbert, 2017). to mainly occur within the middle and deep layers. Its larval
dispersal between the two sides of the Luzon Strait could be
greatly restricted by the effect of the NPIW widely distributed
between a depth range of 300–800 m (Watanabe and Wakatsuchi,
1998), approximately 300 m above the SCS methane seep and
approximately 700 m above the OT hydrothermal vent. The
physical oceanographic modeling results also revealed limited
water connection between these two sites at the middle (500 m)
and deep (1,000 m) layers. Local Adaptation of Shinkaia crosnieri In this process, metalloreductase
(STEAP3), an endosomal membrane enzyme, can convert iron
from an insoluble ferric (Fe3+) to a soluble ferrous (Fe2+) form
(Martinez-Finley et al., 2012). Then, Fe2+ is transported directly
from the endosome to mitochondria (Paul et al., 2017). STEAP3
was unigene upregulated in the SCS seep population, which
may be a strategy against comparatively higher level of dissolved
Fe concentration in the SCS than in the OT (Hu et al., 2015;
Miyazaki et al., 2017a,b). Hydrogen sulfide (H2S) is an important energy source
driving carbon fixation and other critical metabolic processes
in deep-sea thiotrophs, which then supply energy and nutrients
to many macrobenthos living in hydrothermal vents and
methane seeps. However, this gas is also a potent toxin
to cytochrome c oxidase. Therefore, animals living in these
chemosynthesis-based ecosystems are expected to be able to
detoxify sulfide (Julian et al., 2005). Organisms inhabiting H2S-
rich environments have evolved many strategies for coping with
continuous exposure to H2S (Tobler et al., 2016). Eukaryotes
usually possess a mitochondrial sulfur detoxification pathway,
which converts sulfide into sulfite. Two key enzymes in the
sulfide detoxification system, sulfide:quinone oxidoreductase
(SQR) and sulfur dioxygenase (SDO), have been identified in
this study. SQR is a mitochondrial membrane-bound enzyme. It converts sulfide to persulfides and transfers electrons to
the pool of ubiquinone, which is then oxidized by SDO to
generate sulfite with the consumption of molecular oxygen
and persulfide molecules (Hildebrandt and Grieshaber, 2010). A previous study of the mussel G. platifrons from the SCS
methane seep discovered the expression of the same sulfide
detoxification pathway in the symbiont-hosting gill (Wong et al.,
2015; Sun et al., 2017). Succinate dehydrogenase [ubiquinone]
iron-sulfur subunit (SDHB), a subunit of SQR, was identified Plastin is involved in the formation of cross-linked actin
filaments, which are also responsible for the structural solidity
of certain cells (Shinomiya, 2012). Plastin is a protein known for
binding metals including Ca and could therefore be a candidate
for binding uranium (U) with a low pH of 5.2 (Bucher et al.,
2016). If the alteration in plastin occurs in the presence of U,
it may contribute to the structural damage previously observed
in gill tissues of zebrafish (Barillet et al., 2010). Hu et al. Frontiers in Marine Science | www.frontiersin.org Local Adaptation of Shinkaia crosnieri (2015)
reported that cold seeps of the northern SCS contained enriched
authigenic U in sediments, which may explain its influence on November 2020 | Volume 7 | Article 587686 Frontiers in Marine Science | www.frontiersin.org 11 Population Connectivity and Deep-Sea Adaptation Xiao et al. TABLE 6 | Comparison of DEGs involved in antioxidation and detoxification, heat shock proteins, and immune defense with those in the study of Cheng et al. (2020). Gene category
Gene ID
Expression level
Fold
change
FDR
Expression
pattern
Annotation
SCS
OT
Antioxidation and detoxification
Peroxidase
TRINITY_DN10835_c0_g1_i3
49.46
11.73
4.22
3.77E-04
O
Peroxidase
TRINITY_DN66309_c0_g1_i4
24.88
7.16
3.47
9.40E-05
O
Peroxidase
CSE
TRINITY_DN2459_c1_g1_i4
322.20
819.52
0.39
6.68E-15
S
Cystathionine gamma-lyase
DDH
TRINITY_DN4130_c0_g1_i1
21.70
1.49
14.60
1.17E-16
O
Trans-1,2-dihydrobenzene-1,2-diol
dehydrogenase
GST
TRINITY_DN42396_c0_g2_i1
3806.61
524.57
7.26
3.85E-11
O
Glutathione S-transferase Mu 5
Heat Shock Proteins
HSP
TRINITY_DN3174_c0_g2_i5
135.72
315.51
0.43
2.36E-02
S
Heat shock protein 22
Immune defense
CTL
TRINITY_DN57189_c0_g1_i1
160.49
13.34
12.03
3.59E-04
C-type lectin containing low-density lipoprotein
receptor domain
TRINITY_DN3217_c0_g1_i6
278.21
30.75
9.05
2.41E-38
C-type lectin domain family 10 member A
TRINITY_DN95076_c0_g1_i1
5.89
0.72
8.13
2.89E-02
C-type lectin superfamily 17 member A
TRINITY_DN6868_c0_g1_i12
4.47
0.87
5.17
1.90E-05
C-type lectin domain family 4 member M-like
TRINITY_DN5535_c0_g1_i1
14.74
3.56
4.14
1.62E-05
C-type lectin 37Db
TRINITY_DN17218_c0_g1_i7
199.88
76.03
2.63
1.45E-02
C-type lectin containing low-density lipoprotein
receptor domain
TRINITY_DN11643_c0_g1_i2
13.63
36.57
0.37
3.32E-07
C-type lectin domain family 4 member M-like
TRINITY_DN19395_c0_g1_i3
1.26
3.79
0.33
8.46E-05
C-type lectin domain family 4 member C-like
TRINITY_DN799_c0_g1_i4
7.02
22.12
0.32
1.04E-02
C-type lectin domain family 4 member M
TRINITY_DN347_c1_g1_i1
99.98
482.65
0.21
2.05E-09
C-type lectin
TRINITY_DN3728_c0_g1_i2
5.09
34.93
0.15
2.19E-04
C-type lectin domain family 17, member A
TRINITY_DN3401_c0_g1_i3
0.49
187.91
0.00
6.68E-03
C-type lectin
TFPI
TRINITY_DN1673_c0_g2_i1
6700.80
1805.69
3.71
6.90E-04
O
Tissue factor pathway inhibitor 2
Serine protease
TRINITY_DN1726_c0_g1_i5
179.85
22.97
7.83
2.89E-03
O
Serine proteinase stubble
TRINITY_DN4588_c0_g1_i2
22.85
48.87
0.47
5.42E-03
S
Serine proteinase stubble
TRINITY_DN62208_c0_g1_i2
2.20
5.96
0.37
6.35E-03
S
Serine proteinase stubble
Ig
TRINITY_DN27402_c1_g1_i4
2.62
10.60
0.25
4.19E-03
S
Immunoglobulin domain-containing protein
oig-4
A2M
TRINITY_DN3_c0_g1_i7
2170.77
704.60
3.08
4.78E-02
S
Alpha-2-macroglobulin
Expression pattern: S, same/similar; O, opposite. DEGs from SCS population are bold. Ig study of gene expression profiling to prognose lymphoma, breast
cancer, head cancer, and neck cancer, all data sets showed that
predictive inaccuracy was reduced by increasing the sample
size (Dunkler et al., 2007). Local Adaptation of Shinkaia crosnieri Our empirical experience showed
that, based on ∼3 Gb of 150-bp data per sample, at least
five samples would be required to detect sufficient DEGs for
comparing transcriptomes from two different populations. When
only three replicates were used in a simulation with our data,
only about half of the DEGs were detected. Moreover, when
less data per sample were available (2.3 Gb of 100-bp data)
(Cheng et al., 2019), even fewer DEGs (545) could be detected. Our results are consistent with the results of another study that
indicated at least six replicates should be used in order to obtain
sufficient DEGs for a study of the treatment effects (Schurch
et al., 2016). Meanwhile, our subsampling indicated the need to
choose up-to-date methodology in order to detect transcriptome-
wide SNPs. Our empirical results will thus be useful when
planning population genetic studies of not only the squat lobster
S. crosnieri, but also other invertebrates. the genetic variation of plastin. Chitin deacetylases may convert
chitin into chitosan by enzymatic deacetylation of the amino
group of chitin (Raval et al., 2017). Chitin and chitosan both
play a role in metal ions absorption (Kurita, 2006). Previous
study found metal ions such as Sr2+, Mg2+, Na+, Ca2+, and K+
simulate the activity rate of CDA and further inhibited by Co2+,
Ba2+ (Chai et al., 2020). Trace metals such as Sr2+, Mg2+, and
Ca2+ have been detected in the seawater of both the SCS seep and
OT vent sites, with Sr2+ being more abundant in the SCS site than
in the OT site (Hu et al., 2015; Miyazaki et al., 2017a,b). Plastin-2
(PLSL) and chitin deacetylase 1 (CDA1) were enrichened under
GO term from the outlier SNP dataset. CONCLUSION of DEGs between the two populations. Altogether, our study
provide new insights into the genetic structure of S. crosnieri and
molecular mechanisms that have enabled it to thrive in both vents
and seeps. A broader sampling involving more localities with
contrasting environmental conditions including both different
temperatures and depths is desired in the future to further
improve our understanding of the population dynamics and local
adaptation of this widespread species in the Western Pacific. Using transcriptome sequencing, we identified DEGs and PSGs
between two Shinkaia crosnieri populations, one from a methane
seep in the SCS and another from a hydrothermal vent field in
the OT. These genes were found to be involved and enriched
in metabolic pathways including oxidoreductase activity and
sulfur metabolism that may be important in adaptation to
different environmental conditions. Using transcriptome-wide
SNP markers, we confirmed clear genetic subdivision between
S. crosnieri from the SCS and the OT, and suggested the Luzon
Strait as an important physical barrier to the larval dispersal
between the two populations. Moreover, our outlier analysis
revealed the involvement of genes related to metal ion binding
and energy metabolism in shaping the genetic divergence of the
two populations. Using a subsampling strategy, we found that at
least five replicates were required to capture the full spectrum FIGURE 6 | (A) Number of DEGs identified based on the subsampling approach by random selecting 3 (T3), 5 (T5), 10 (T10), and 15 (T15) individuals from each
local population and comparing with Cheng et al. (2019), with yellow, red, and blue dots indicating DEGs in SCS, OT, and total DEGs number, respectively. (B) Number of SNPs identified based on the subsampling approach by random selecting 3 (T3), 5 (T5), and 10 (T10) individuals from each local population and
comparison with Cheng et al. (2020). FIGURE 6 | (A) Number of DEGs identified based on the subsampling approach by random selecting 3 (T3), 5 (T5), 10 (T10), and 15 (T15) individuals from each
local population and comparing with Cheng et al. (2019), with yellow, red, and blue dots indicating DEGs in SCS, OT, and total DEGs number, respectively. (B) Number of SNPs identified based on the subsampling approach by random selecting 3 (T3), 5 (T5), and 10 (T10) individuals from each local population and
comparison with Cheng et al. (2020). Frontiers in Marine Science | www.frontiersin.org Sample Size Advantages Transcriptome sequencing data are able to reveal gene expression
patterns in different sample tissues or in samples under different
conditions. Nevertheless, an appropriate sample size should be
applied for transcriptome-based studies as it will affect the
results obtained (Aach and Church, 2001). For example, in a November 2020 | Volume 7 | Article 587686 Frontiers in Marine Science | www.frontiersin.org 12 Population Connectivity and Deep-Sea Adaptation Xiao et al. Xiao et al. A
B
FIGURE 6 | (A) Number of DEGs identified based on the subsampling approach by random selecting 3 (T3), 5 (T5), 10 (T10), and 15 (T15) individuals from each
local population and comparing with Cheng et al. (2019), with yellow, red, and blue dots indicating DEGs in SCS, OT, and total DEGs number, respectively. (B) Number of SNPs identified based on the subsampling approach by random selecting 3 (T3), 5 (T5), and 10 (T10) individuals from each local population and
comparison with Cheng et al. (2020). A REFERENCES Bradnam, K. R., Fass, J. N., Alexandrov, A., Baranay, P., Bechner, M., Birol,
I., et al. (2013). Assemblathon 2: evaluating de novo methods of genome
assembly in three vertebrate species. GigaScience 2:10. doi: 10.1186/2047-21
7X-2-10 Aach, J., and Church, G. M. (2001). Aligning gene expression time series with time
warping algorithms. Bioinformatics 17, 495–508. doi: 10.1093/bioinformatics/
17.6.495 Bray, N. L., Pimentel, H., Melsted, P., and Pachter, L. (2016). Near-optimal
probabilistic RNA-seq quantification. Nat. Biotechnol. 34:525. doi: 10.1038/nbt. 3519 Alexander, D. H., Novembre, J., and Lange, K. (2009). Fast model-based estimation
of ancestry in unrelated individuals. Genome Res. 19, 1655–1664. doi: 10.1101/
gr.094052.109 Bucher, G., Mounicou, S., Simon, O., Floriani, M., Lobinski, R., and Frelon, S. (2016). Insights into the nature of uranium target proteins within zebrafish
gills after chronic and acute waterborne exposures. Environ. Toxicol. Chem. 35,
736–741. doi: 10.1002/etc.3249 Alexander,
D. H.,
Shringarpure,
S. S.,
Novembre,
J.,
and
Lange,
K. (2015). Admixture
1.3
Software
Manual. Available
online
at:
https://www.genetics.ucla.edu/software/admixture/admixture-manual.pdf
(accessed March 23, 2020). Chai, J., Hang, J., Zhang, C., Yang, J., Wang, S., Liu, S., et al. (2020). Purification
and characterization of chitin deacetylase active on insoluble chitin from
Nitratireductor aquimarinus MCDA3-3. Int. J. Biol. Macromol. 152, 922-929. doi: 10.1016/j.ijbiomac.2020.02.308 Altschul, S. F., Gish, W., Miller, W., Myers, E. W., and Lipman, D. J. (1990). Basic
local alignment search tool. J. Mol. Biol. 215, 403–410. doi: 10.1016/S0022-
2836(05)80360-2 Chan, T. Y., Lee, D. A., and Lee, C. S. (2000). The first deep-sea hydrothermal
animal reported from Taiwan: Shinkaia crosnieri Baba and Williams, 1998
(Crustacea: Decapoda: Galatheidae). Bull. Mar. Sci. 67, 799–804. Arellano, S. M., and Young, C. M. (2009). Spawning, development, and the
duration of larval life in a deep-sea cold-seep mussel. Biol. Bull. 216, 149–162. doi: 10.1086/BBLv216n2p149 Baba, K., and Williams, A. (1998). New galatheoidea (Crustacea, Decapoda,
Anomura) from hydrothermal systems in the West Pacific Ocean: bismarck
archipelago and Okinawa Trough. Zoosystema 20, 143–156. doi: 10.11646/
zootaxa.4057.1.5 Chen, E. Y., Tan, C. M., Kou, Y., Duan, Q., Wang, Z., Meirelles, G. V., et al. (2013). Enrichr: interactive and collaborative HTML5 gene list enrichment analysis
tool. BMC Bioinform. 14:128. doi: 10.1186/1471-2105-14-128 Cheng, J., Hui, M., Li, Y., and Sha, Z. (2020). Genomic evidence of
population genetic differentiation in deep-sea squat lobster Shinkaia crosnieri
(Crustacea: Decapoda: Anomura) from Northwestern Pacific hydrothermal
vent and cold seep. Oceanogr. Res. Pap. 156:103188. ETHICS STATEMENT (GML2019ZD0409) as well as by a grant from China Ocean
Mineral Resources Research and Development Association
(DY135-E2-1-03). The R/V Kairei research cruise KR15-17 (PI:
Hiroyuki Yamamoto) was supported by the Council for Science,
Technology, and Innovation (CSTI) in Japan as part of the
Cross Ministerial Strategic Innovation Promotion Program (SIP),
Next-generation Technology for Ocean Resource Exploration. The animal study was reviewed and approved by The Ethics
Committee of the Hong Kong University of Science and
Technology. Written informed consent was obtained from the
owners for the participation of their animals in this study. DATA AVAILABILITY STATEMENT The datasets generated from this study can be found in
online repositories. The names of the repository/repositories and
accession number(s) can be found below: https://www.ncbi.nlm. nih.gov/, PRJNA612951. November 2020 | Volume 7 | Article 587686 Frontiers in Marine Science | www.frontiersin.org 13 Population Connectivity and Deep-Sea Adaptation Xiao et al. FUNDING This work was supported by grants from the Major Basic
and
Applied
Research
Projects
of
Guangdong
Province
(2019B030302004-04), the Hong Kong Branch of Southern
Marine
Science
and
Engineering
Guangdong
Laboratory
(Guangzhou)
(SMSEGL20SC01),
and
Southern
Marine
Science and Engineering Guangdong Laboratory (Guangzhou) This work was supported by grants from the Major Basic
and
Applied
Research
Projects
of
Guangdong
Province
(2019B030302004-04), the Hong Kong Branch of Southern
Marine
Science
and
Engineering
Guangdong
Laboratory
(Guangzhou)
(SMSEGL20SC01),
and
Southern
Marine
Science and Engineering Guangdong Laboratory (Guangzhou) This work was supported by grants from the Major Basic
and
Applied
Research
Projects
of
Guangdong
Province
(2019B030302004-04), the Hong Kong Branch of Southern
Marine
Science
and
Engineering
Guangdong
Laboratory
(Guangzhou)
(SMSEGL20SC01),
and
Southern
Marine
Science and Engineering Guangdong Laboratory (Guangzhou) SUPPLEMENTARY MATERIAL The Supplementary Material for this article can be found
online
at:
https://www.frontiersin.org/articles/10.3389/fmars. 2020.587686/full#supplementary-material ACKNOWLEDGMENTS P-YQ and J-WQ conceived and designed the project. J-WQ, JS,
and CC collected the samples. TX and YX extracted the RNA. YX,
YHK, and WCW performed the real-time PCR experiments. YW
conducted ocean modeling analysis. YX, TX, and JS performed
the bioinformatics analyses and drafted the manuscript. All
authors contributed to the manuscript and approved it for
submission and publication. We thank the captains and crews of Xiangyanghong 9, R/V
Kairei, and R/V Tan Ka Kee, as well as the operation team
of the deep-submergence vehicle Jiaolong, ROV KAIKO, and
ROV ROPOS for helping us with sample collection during the
relevant cruises. Hiroyuki Yamamoto (JAMSTEC) is thanked for
serving as the chief scientist of the research cruise KR15-17. The data for physical oceanographic modeling in this study can
be found on the http://www.hycom.org data server under the
“HYCOM + NCODA Global 1/12◦Reanalysis” link. REFERENCES pcadapt: an R package to perform
genome scans for selection based on principal component analysis. Mol. Ecol. Resour. 17, 67–77. doi: 10.1111/1755-0998.12592 Grabherr, M. G., Haas, B. J., Yassour, M., Levin, J. Z., Thompson, D. A., Amit,
I., et al. (2011). Trinity: reconstructing a full-length transcriptome without a
genome from RNA-Seq data. Nat. Biotechnol. 29:644. doi: 10.1038/nbt.1883 Martinez-Finley, E. J., Chakraborty, S., Fretham, S. J., and Aschner, M. (2012). Cellular transport and homeostasis of essential and nonessential metals. Metallomics 4, 593–605. doi: 10.1039/C2MT00185C Ha, B. G., Jung, S. S., Jang, Y. K., Jeon, B. Y., and Shon, Y. H. (2019). Mineral-
enriched deep-sea water modulates lactate metabolism via PGC-1α-mediated
metabolic reprogramming. Mar. Drugs 17:611. doi: 10.3390/md17110611 Miyake, H., Kitada, M., Nakamura, K. I., Omata, T., and Itoh, T. (2010). Larvae of
deep-sea chemosynthetic ecosystem animals in captivity. Cahiers Biol. Mar. 51,
441–450. doi: 10.1111/j.1439-0485.2006.00115.x Haas, B. J., Papanicolaou, A., Yassour, M., Grabherr, M., Blood, P. D., Bowden, J.,
et al. (2013). De novo transcript sequence reconstruction from RNA-seq using
the Trinity platform for reference generation and analysis. Nature protocols 8,
1494–1512. doi: 10.1038/nprot.2013.084 Miyazaki, J., Kawagucci, S., Makabe, A., Takahashi, A., Kitada, K., Torimoto,
J., et al. (2017a). Deepest and hottest hydrothermal activity in the Okinawa
Trough: the Yokosuka site at Yaeyama Knoll. Royal Soc. Open Sci. 4:171570. doi: 10.1098/rsos.171570 Helbert, W. (2017). Marine polysaccharide sulfatases. Front. Mar. Sci. 4:6. doi:
10.3389/fmars.2017.00006 Hildebrandt, T. M., and Grieshaber, M. K. (2010). Three enzymatic activities
catalyze the oxidation of sulfide to thiosulfate in mammalian and invertebrate
mitochondria. FEBS
J. 275,
3352–3361. doi:
10.1111/j.1742-4658.2008. 06482.x Miyazaki, J., Makabe, A., Matsui, Y., Ebina, N., Tsutsumi, S., Ishibashi, J. I.,
et al. (2017b). WHATS-3: an improved flow-through multi-bottle fluid sampler
for deep-sea geofluid research. Front. Earth Sci. 5:45. doi: 10.3389/feart.2017. 00045 Houston, R. D., Taggart, J. B., Cézard, T., Bekaert, M., Lowe, N. R., Downing, A.,
et al. (2014). Development and validation of a high density SNP genotyping
array for Atlantic salmon (Salmo salar). BMC Genom. 15:90. doi: 10.1186/1471-
2164-15-90 Miyazaki, J. I., de Oliveira Martins, L., Fujita, Y., Matsumoto, H., and Fujiwara,
Y. (2010). Evolutionary process of deep-sea Bathymodiolus mussels. PLoS One
5:e10363. doi: 10.1371/journal.pone.0010363 Hu, Y., Feng, D., Liang, Q., Xia, Z., Chen, L., and Chen, D. (2015). Impact of
anaerobic oxidation of methane on the geochemical cycle of redox-sensitive
elements at cold-seep sites of the northern South China Sea. Top. Stud. Oceanogr. REFERENCES doi: 10.1016/j.ejca.2006.11.018 Lechner, M., Findeiß, S., Steiner, L., Marz, M., Stadler, P. F., and Prohaska, S. J. (2011). Proteinortho: detection of (co-) orthologs in large-scale analysis. BMC
Bioinform. 12:124. doi: 10.1186/1471-2105-12-124 Excoffier, L., and Lischer, H. E. (2010). Arlequin suite ver 3.5: a new series of
programs to perform population genetics analyses under Linux and Windows. Mol. Ecol. Resour. 10, 564–567. doi: 10.1111/j.1755-0998.2010.02847.x Leclerc, C., Haeich, J., Aulestia, F. J., Kilhoffer, M. C., Miller, A. L., Neant, I., et al. (2016). Calcium signaling orchestrates glioblastoma development: facts and
conjunctures. Biochim. Biophys. Acta 1863, 1447–1459. doi: 10.1016/j.bbamcr. 2016.01.018 Fu, L., Niu, B., Zhu, Z., Wu, S., and Li, W. (2012). CD-HIT: accelerated for
clustering the next-generation sequencing data. Bioinformatics 28, 3150–3152. doi: 10.1093/bioinformatics/bts565 Lee, R. F. (1981). Mixed function oxygenases (MFO) in marine invertebrates. Mar. Biol. Lett. 2, 87–105. German, C., Livermore, R., Baker, E., Bruguier, N., Connelly, D., Cunningham, A.,
et al. (2000). Hydrothermal plumes above the East Scotia Ridge: an isolated
high-latitude back-arc spreading centre. Earth Planet. Sci. Lett. 184, 241–250. doi: 10.1016/S0012-821X(00)00319-8 Levin, L. A. (2005). “Ecology of cold seep sediments: interactions of fauna with
flow, chemistry and microbes,” in Oceanography and Marine Biology: An Annual
Review, eds R. Gibson, R. Atkinson, and J. Gordon (Boca Raton, FL: Taylor and
French), 1–46. doi: 10.1201/9781420037449.ch1 German, C. R., Ramirez-Llodra, E., Baker, M. C., Tyler, P. A., and Committee,
C. S. S. (2011). Deep-water chemosynthetic ecosystem research during the
census of marine life decade and beyond: a proposed deep-ocean road map. PLoS One 6:e23259. doi: 10.1371/journal.pone.0023259 Li, H. (2011). A statistical framework for SNP calling, mutation discovery,
association mapping and population genetical parameter estimation from
sequencing data. Bioinformatics 27, 2987–2993. doi: 10.1093/bioinformatics/
btr509 Gobe, G., and Crane, D. (2010). Mitochondria, reactive oxygen species and
cadmium toxicity in the kidney. Toxicol. Lett. 198, 49–55. doi: 10.1016/j.toxlet. 2010.04.013 Li, H., Handsaker, B., Wysoker, A., Fennell, T., Ruan, J., Homer, N., et al. (2009). The sequence alignment/map format and SAMtools. Bioinformatics 25, 2078–
2079. doi: 10.1093/bioinformatics/btp352 Götz, S., García-Gómez, J. M., Terol, J., Williams, T. D., Nagaraj, S. H., Nueda, M. J.,
et al. (2008). High-throughput functional annotation and data mining with the
Blast2GO suite. Nucl. Acids Res. 36, 3420–3435. doi: 10.1093/nar/gkn176 Love, M., Anders, S., and Huber, W. (2014). Differential analysis of count data–the
DESeq2 package. Genome Biology 15:550. doi: 10.1186/s13059-014-0550-8 Luu, K., Bazin, E., and Blum, M. G. (2017). REFERENCES doi: 10.1016/j.dsr.2019.10
3188 Barillet, S., Larno, V., Floriani, M., Devaux, A., and Adam-Guillermin, C. (2010). Ultrastructural effects on gill, muscle, and gonadal tissues induced in zebrafish
(Danio rerio) by a waterborne uranium exposure. Aquat. Toxicol. 100, 295–302. doi: 10.1016/j.aquatox.2010.08.002 Cheng, J., Hui, M., and Sha, Z. (2019). Transcriptomic analysis reveals insights into
deep-sea adaptations of the dominant species, Shinkaia crosnieri (Crustacea:
Decapoda: Anomura), inhabiting both hydrothermal vents and cold seeps. BMC
Genom. 20:388. doi: 10.1186/s12864-019-5753-7 Belcaid, M., Casaburi, G., McAnulty, S. J., Schmidbaur, H., Suria, A. M., Moriano-
Gutierrez, S., et al. (2019). Symbiotic organs shaped by distinct modes of
genome evolution in cephalopods. Proc. Natl. Acad. Sci. U.S.A. 116, 3030–3035. doi: 10.1073/pnas.1817322116 Bolger, A. M., Lohse, M., and Usadel, B. (2014). Trimmomatic: a flexible
trimmer for Illumina sequence data. Bioinformatics 30, 2114–2120. doi: 10. 1093/bioinformatics/btu170 Corliss, J. B., Dymond, J., Gordon, L. I., Edmond, J. M., von Herzen, R. P., Ballard,
R. D., et al. (1979). Submarine thermal springs on the Galapagos Rift. Science
203, 1073–1083. doi: 10.1126/science.203.4385.1073 November 2020 | Volume 7 | Article 587686 Frontiers in Marine Science | www.frontiersin.org 14 Population Connectivity and Deep-Sea Adaptation Xiao et al. Kiel, S. (2016). A biogeographic network reveals evolutionary links between deep-
sea hydrothermal vent and methane seep faunas. Biol. Sci. 283:20162337. doi:
10.1098/rspb.2016.2337 Cromer, B. A., Gorman, M. A., Hansen, G., Adams, J. J., Coggan, M., Board, P. G.,
et al. (2007). Expression, purification, crystallization and preliminary X-ray
diffraction analysis of chloride intracellular channel 2 (CLIC2). Struct. Biol. Crystall. Commun. 63, 961–963. doi: 10.1107/S1744309107049159 Kurita, K. (2006). Chitin and chitosan: functional biopolymers from marine
crustaceans. Mar. Biotechnol. 8, 203–226. doi: 10.1007/s10126-005-0097-5 Danecek, P., Auton, A., Abecasis, G., Albers, C. A., Banks, E., DePristo, M. A., et al. (2011). The variant call format and VCFtools. Bioinformatics 27, 2156–2158. doi: 10.1093/bioinformatics/btr330 Kyuno, A., Shintaku, M., Fujita, Y., Matsumoto, H., Utsumi, M., Watanabe, H.,
et al. (2009). Dispersal and differentiation of deep-sea mussels of the genus
Bathymodiolus (Mytilidae. Bathymodiolinae). J. Mar. Biol. 2009, 1–15. doi:
10.1155/2009/625672 Deponte, M. (2013). Glutathione catalysis and the reaction mechanisms of
glutathione-dependent enzymes. Biochim. Biophys. Acta 1830, 3217–3266. doi:
10.1016/j.bbagen.2012.09.018 Langmead, B., and Salzberg, S. L. (2012). Fast gapped-read alignment with Bowtie
2. Nat. Methods 9, 357–359. doi: 10.1038/nmeth.1923 Dunkler, D., Michiels, S., and Schemper, M. (2007). Gene expression profiling: does
it add predictive accuracy to clinical characteristics in cancer prognosis? Eur. J. Cancer 43, 745–751. REFERENCES 122, 84–94. doi: 10.1016/j.dsr2.2015.06.012 Nakajima, Y., Shinzato, C., Khalturina, M., Nakamura, M., Watanabe, H. K.,
Nakagawa, S., et al. (2018). Isolation and characterization of novel polymorphic
microsatellite loci for the deep-sea hydrothermal vent limpet, Lepetodrilus nux,
and the vent-associated squat lobster, Shinkaia crosnieri. Mar. Biodivers. 48,
677–684. doi: 10.1007/s12526-017-0704-5 Julian, D., April, K. L., Patel, S., Stein, J. R., and Wohlgemuth, S. E. (2005). Mitochondrial depolarization following hydrogen sulfide exposure in
erythrocytes from a sulfide-tolerant marine invertebrate. J. Exp. Biol. 208,
4109–4122. doi: 10.1242/jeb.01867 Nakamura, K., Kawagucci, S., Kitada, K., Kumagai, H., Takai, K., and Okino, K. (2015). Water column imaging with multibeam echo-sounding in the mid-
Okinawa Trough: implications for distribution of deep-sea hydrothermal vent
sites and the cause of acoustic water column anomaly. Geochem. J. 49, 579–596. doi: 10.2343/geochemj.2.0387 Keenan, K., McGinnity, P., Cross, T. F., Crozier, W. W., and Prodöhl, P. A. (2013). diveRsity: an R package for the estimation and exploration of population
genetics parameters and their associated errors. Methods Ecol. Evol. 4, 782–788. doi: 10.1111/2041-210X.12067 Nan, F., Xue, H., Chai, F., Shi, L., Shi, M., and Guo, P. (2011). Identification of
different types of Kuroshio intrusion into the South China Sea. Ocean Dyn. 61,
1291–1304. doi: 10.1007/s10236-011-0426-3 Kendrick-Jones, J., Szentkiralyi, E. M., and Szent-Györgyi, A. G. (1976). Regulatory
light chains in myosins. J. Mol. Biol. 104, 747–775. doi: 10.1016/0022-2836(76)
90180-7 Parra, E. J., Marcini, A., Akey, J., Martinson, J., Batzer, M. A., Cooper, R.,
et al. (1998). Estimating African American admixture proportions by use of November 2020 | Volume 7 | Article 587686 Frontiers in Marine Science | www.frontiersin.org 15 Population Connectivity and Deep-Sea Adaptation Xiao et al. population-specific alleles. Am. J. of Hum. Genet. 63, 1839–1851. doi: 10.1086/
302148 population-specific alleles. Am. J. of Hum. Genet. 63, 1839–1851. doi: 10.1086/
302148 Trimukhe, K. D., and Varma, A. J. (2008). Complexation of heavy metals by
crosslinked chitin and its deacetylated derivatives. Carbohydr. Polym. 71, 66–73. doi: 10.1016/j.carbpol.2007.05.016 Paul, B. T., Manz, D. H., Torti, F. M., and Torti, S. V. (2017). Mitochondria and
Iron: current questions. Exp. Rev. Hematol. 10, 65–79. doi: 10.1080/17474086. 2016.1268047 Van Dover, C. L., German, C., Speer, K. G., Parson, L., and Vrijenhoek, R. (2002). Evolution and biogeography of deep-sea vent and seep invertebrates. Science
295, 1253–1257. doi: 10.1126/science.1067361 Pegadaraju, V., Nipper, R., Hulke, B., Qi, L., and Schultz, Q. (2013). REFERENCES De novo
sequencing of sunflower genome for SNP discovery using RAD (Restriction
site Associated DNA) approach. BMC Genom. 14:556. doi: 10.1186/1471-2164-
14-556 Vrijenhoek, R. C. (2013). On the instability and evolutionary age of deep-sea
chemosynthetic communities. Top. Stud. Oceanogr. 92, 189–200. doi: 10.1016/
j.dsr2.2012.12.004 Watanabe, H., Fujikura, K., Kojima, S., Miyazaki, J. I., and Fujiwara, Y. (2010). “Japan: Vents and Seeps in Close Proximity,” in The vent and seep
biota, ed. S. Kiel (Dordrecht: Springer), 379–401. doi: 10.1007/978-90-481-95
72-5_12 Purcell, S., Neale, B., Todd-Brown, K., Thomas, L., Ferreira, M. A., Bender, D.,
et al. (2007). PLINK: a tool set for whole-genome association and population-
based linkage analyses. Am. J. Hum. Genet. 81, 559–575. doi: 10.1086/51
9795 Watanabe, T., and Wakatsuchi, M. (1998). Formation of 26.8–26.9 σθ water in the
kuril basin of the sea of okhotsk as a possible origin of north pacific intermediate
water. J. Geophys. Res. 103, 2849–2865. doi: 10.1029/97JC02914 Ramos, R., and Garcia, E. (2007). Induction of mixed-function oxygenase system
and antioxidant enzymes in the coral Montastraea faveolata on acute exposure
to benzo (a) pyrene. Toxicol. Pharmacol. 144, 348–355. doi: 10.1016/j.cbpc.2006. 11.006 Wong, Y. H., Sun, J., He, L. S., Chen, L. G., Qiu, J. W., and Qian, P. Y. (2015). High-
throughput transcriptome sequencing of the cold seep mussel Bathymodiolus
platifrons. Sci. Rep. 5:16597. doi: 10.1038/srep16597 Raval, R., Simsa, R., and Raval, K. (2017). Expression studies of Bacillus
licheniformis chitin deacetylase in E. coli Rosetta cells. Int. J. Biol. Macromol. 104, 1692–1696. doi: 10.1016/j.ijbiomac.2017.01.151 Xu, J., Ji, P., Zhao, Z., Zhang, Y., Feng, J., Wang, J., et al. (2012). Genome-wide SNP
discovery from transcriptome of four common carp strains. PLoS One 7:e48140. doi: 10.1371/journal.pone.0048140 Robinson, M. D., McCarthy, D. J., and Smyth, G. K. (2010). edgeR: a Bioconductor
package for differential expression analysis of digital gene expression data. Bioinformatics 26, 139–140. doi: 10.1093/bioinformatics/btp616 Xu, T., Sun, J., Lv, J., Watanabe, H. K., Li, T., Zou, W., et al. (2017). Genome-wide
discovery of single nucleotide polymorphisms (SNPs) and single nucleotide
variants (SNVs) in deep-sea mussels: Potential use in population genomics and
cross-species application. Top. Stud. Oceanogr. 137, 318–326. doi: 10.1016/j. dsr2.2016.03.011 Rubin-Blum, M., Antony, C. P., Borowski, C., Sayavedra, L., Pape, T., Sahling,
H., et al. (2017). Short-chain alkanes fuel mussel and sponge Cycloclasticus
symbionts from deep-sea gas and oil seeps. Nature microbiology 2:17093. doi:
10.1038/nmicrobiol.2017.93 Schurch, N. REFERENCES J., Schofield, P., Gierli´nski, M., Cole, C., Sherstnev, A., Singh, V., et al. (2016). How many biological replicates are needed in an RNA-seq experiment
and which differential expression tool should you use? RNA 22, 839–851. doi:
10.1261/rna.053959.115 Xu, T., Sun, J., Watanabe, H. K., Chen, C., Nakamura, M., Ji, R., et al. (2018). Population genetic structure of the deep-sea mussel Bathymodiolus platifrons
(Bivalvia: Mytilidae) in the Northwest Pacific. Evol. Appl. 11, 1915–1930. doi:
10.1111/eva.12696 Shen, Y., Kou, Q., Chen, W., He, S., Yang, M., Li, X., et al. (2016). Comparative population structure of two dominant species, Shinkaia
crosnieri (Munidopsidae: Shinkaia) and Bathymodiolus platifrons (Mytilidae:
Bathymodiolus), inhabiting both deep-sea vent and cold seep inferred from
mitochondrial multi-genes. Ecol. Evol. 6, 3571–3582. doi: 10.1002/ece3. 2132 Yahagi, T., Watanabe, H., Kojima, S., and Kano, Y. (2017). Do larvae from deep-
sea hydrothermal vents disperse in surface waters? Ecology 98, 1524–1534. doi: 10.1002/ecy.1800 Yang, C. H., Tsuchida, S., Fujikura, K., Fujiwara, Y., Kawato, M., and Chan,
T. Y. (2016). Connectivity of the squat lobsters Shinkaia crosnieri (Crustacea:
Decapoda: Galatheidae) between cold seep and hydrothermal vent habitats. Bull. Mar. Sci. 92, 17–31. doi: 10.5343/bms.2015.1031 Shinomiya, H. (2012). Plastin family of actin-bundling proteins: its functions in
leukocytes, neurons, intestines, and cancer. Int. J. Cell Biol. 2012, 213492. doi:
10.1155/2012/213492 You, Y., Chern, C. S., Yang, Y., Liu, C. T., Liu, K. K., and Pai, S. C. (2005). The
South China Sea, a cul-de-sac of North Pacific intermediate water. J. Oceanogr. 61, 509–527. doi: 10.1007/s10872-005-0059-6 Simão, F. A., Waterhouse, R. M., Ioannidis, P., Kriventseva, E. V., and Zdobnov,
E. M. (2015). BUSCO: assessing genome assembly and annotation completeness
with single-copy orthologs. Bioinformatics 31, 3210–3212. doi: 10.1093/
bioinformatics/btv351 Yuan, Y., Liao, G., Wang, H., Lou, R., and Chen, H. (2009). Variability of the
currents in the Luzon Strait during spring of 2002 obtained from observations
and satellite geostrophic currents and spectral analyses. Earth Sci. 52, 519–531. doi: 10.1007/s11430-009-0041-z Sun, J., Zhang, Y., Xu, T., Zhang, Y., Mu, H., Zhang, Y., et al. (2017). Adaptation
to deep-sea chemosynthetic environments as revealed by mussel genomes. Nat. Ecol. Evol. 1:121. doi: 10.1038/s41559-017-0121 Zhang, Z., Zhang, Y., Evans, P., Chinwalla, A., and Taylor, D. (2017). RNA-seq
2G: online analysis of differential gene expression with comprehesive options
of statistical methods. bioRxiv[Preprint]. doi: 10.1101/122747 Suzuki,
S.,
and
Strominger,
J. L. (1960). Enzymatic
sulfation
of
mucopolysaccharides
in
hen
oviduct
I. REFERENCES Transfer
of
sulfate
from
3’-
phosphoadenosine 5’-phosphosulfate to mucopolysaccharides. J. Biol. Chem. 235, 257–266. Zheng, X., Levine, D., Shen, J., Gogarten, S. M., Laurie, C., and Weir, B. S. (2012). A high-performance computing toolset for relatedness and principal
component analysis of SNP data. Bioinformatics 28, 3326–3328. doi: 10.1093/
bioinformatics/bts606 Swingle, M. R., and Honkanen, R. E. (2019). Inhibitors of serine/threonine
protein phosphatases: biochemical and structural studies provide insight
for further development. Curr. Med. Chem. 26, 2634–2660. doi: 10.2174/
0929867325666180508095242 Conflict of Interest: The authors declare that the research was conducted in the
absence of any commercial or financial relationships that could be construed as a
potential conflict of interest. Tian, J., Yang, Q., Liang, X., Xie, L., Hu, D., Wang, F., et al. (2006). Observation
of Luzon Strait transport. Geophys. Res. Lett. 33:L19607. doi: 10.1029/
2006GL026272 Copyright © 2020 Xiao, Xu, Sun, Wang, Wong, Kwan, Chen, Qiu and Qian. This
is an open-access article distributed under the terms of the Creative Commons
Attribution License (CC BY). The use, distribution or reproduction in other forums
is permitted, provided the original author(s) and the copyright owner(s) are credited
and that the original publication in this journal is cited, in accordance with accepted
academic practice. No use, distribution or reproduction is permitted which does not
comply with these terms. Tian, J., Yang, Q., and Zhao, W. (2009). Enhanced diapycnal mixing in the South
China Sea. J. Phys. Oceanogr. 39, 3191–3203. doi: 10.1175/2009JPO3899.1 Tobler, M., Passow, C. N., Greenway, R., Kelley, J. L., and Shaw, J. H. (2016). The evolutionary ecology of animals inhabiting hydrogen sulfide–rich
environments. Annu. Rev. Ecol. Evol. Syst. 47, 239–262. doi: 10.1146/annurev-
ecolsys-121415-032418 November 2020 | Volume 7 | Article 587686 Frontiers in Marine Science | www.frontiersin.org 16
|
https://openalex.org/W4387399891
|
https://bg.copernicus.org/preprints/bg-2023-176/bg-2023-176.pdf
|
English
| null |
Comment on bg-2023-176
| null | 2,023
|
cc-by
| 19,891
|
Abstract. CO2 emissions exhibited a clear
diurnal pattern, but the exact shape and timing of this pattern differed between habitats. In contrast, CH4 emissions did not
change diurnally. Our data confirm our hypothesis that spatial variability is important for CH4 while diurnal variability is more while 10% of emissions was from dry fallen sediment at the side of the river. Aquatic CO2 emissions were similar at different
20
habitats, while aquatic CH4 emissions were higher at the side of the river. Artificial structures to imoprove navigability
(groynes) created still water areas with elevated CH4 emissions and lower CO2 emissions. CO2 emissions exhibited a clear
diurnal pattern, but the exact shape and timing of this pattern differed between habitats. In contrast, CH4 emissions did not
change diurnally. Our data confirm our hypothesis that spatial variability is important for CH4 while diurnal variability is more while 10% of emissions was from dry fallen sediment at the side of the river. Aquatic CO2 emissions were similar at different
20
habitats, while aquatic CH4 emissions were higher at the side of the river. Artificial structures to imoprove navigability
(groynes) created still water areas with elevated CH4 emissions and lower CO2 emissions. CO2 emissions exhibited a clear
diurnal pattern, but the exact shape and timing of this pattern differed between habitats. In contrast, CH4 emissions did not
change diurnally. Our data confirm our hypothesis that spatial variability is important for CH4 while diurnal variability is more relevant for CO2 emissions. Continuous measurements are most likely necessary for reliable quantification of river GHG
25
emissions. relevant for CO2 emissions. Continuous measurements are most likely necessary for reliable quantification of river GHG
25
emissions. Diurnal versus spatial variability of greenhouse gas emissions from an
anthropogenic modified German lowland river Norbert Kamjunke2, Uta Koedel3, Michael Rode4, Claudia Schuetze3, Ingeborg Matthias Koschorreck1, Norbert Kamjunke2, Uta Koedel3, Michael Rode4, Claudia Schuetze3, Ingeborg
Bussmann5 1Department Lake Research, Helmholtz Centre for Environmental Research, Magdeburg, Germany
5
2Department River Ecology, Helmholtz Centre for Environmental Research, Magdeburg, Germany
3Department Monitoring & Exploration Technologies, Helmholtz Centre for Environmental Research, Leipzig, Germany
4Department Aquatic Ecosystem Analysis, Helmholtz Centre for Environmental Research, Magdeburg, Germany
5Department Shelf Sea System Ecology, Alfred Wegener Institut, Helmholtz Zentrum für Polar- und Meeresforschung,
Helgoland, Germany
10 1Department Lake Research, Helmholtz Centre for Environmental Research, Magdeburg, Germany
5
2Department River Ecology, Helmholtz Centre for Environmental Research, Magdeburg, Germany
3Department Monitoring & Exploration Technologies, Helmholtz Centre for Environmental Research, Leipzig, Germany
4Department Aquatic Ecosystem Analysis, Helmholtz Centre for Environmental Research, Magdeburg, Germany
5Department Shelf Sea System Ecology, Alfred Wegener Institut, Helmholtz Zentrum für Polar- und Meeresforschung,
Helgoland, Germany
10 Correspondence to: Matthias Koschorreck (matthias.koschorreck@ufz.de) https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. Abstract. Greenhous gas (GHG) emissions from rivers are globally relevant, but quantification of these emissions comes with
considerable uncertainty. Most of the existing studies were carried out at small streams and much less is known about GHG
emissions from larger rivers. Here quantification of ecosystem scale emissions is challenged by both spatial and short-term
15
temporal variability. We measured spatio-temporal variability of CO2 and CH4 emissions from a 1 km long reach of the German
lowland river Elbe over three days in order to establish which factor is more relevant to be taken into consideration: small-
scale spatial variability or short-term temporal variability of CO2 and CH4 emissions. g
g
y
q
considerable uncertainty. Most of the existing studies were carried out at small streams and much less is known about GHG
emissions from larger rivers. Here quantification of ecosystem scale emissions is challenged by both spatial and short-term
15
temporal variability. We measured spatio-temporal variability of CO2 and CH4 emissions from a 1 km long reach of the German
lowland river Elbe over three days in order to establish which factor is more relevant to be taken into consideration: small-
scale spatial variability or short-term temporal variability of CO2 and CH4 emissions. GHG emissions from the river reach studied were dominated by CO2 and 90 % of total emissions was from the water surface y
g
emissions from larger rivers. Here quantification of ecosystem scale emissions is challenged by both spatial and short-term
15
temporal variability. We measured spatio-temporal variability of CO2 and CH4 emissions from a 1 km long reach of the German
lowland river Elbe over three days in order to establish which factor is more relevant to be taken into consideration: small-
scale spatial variability or short-term temporal variability of CO2 and CH4 emissions. GHG emissions from the river reach studied were dominated by CO2 and 90 % of total emissions was from the water surface 15 GHG emissions from the river reach studied were dominated by CO2 and 90 % of total emissions was from the water surface,
while 10% of emissions was from dry fallen sediment at the side of the river. Aquatic CO2 emissions were similar at different
20
habitats, while aquatic CH4 emissions were higher at the side of the river. Artificial structures to imoprove navigability
(groynes) created still water areas with elevated CH4 emissions and lower CO2 emissions. 1Department Lake Research, Helmholtz Centre for Environmental Research, Magdeburg, Germany
5
2Department River Ecology, Helmholtz Centre for Environmental Research, Magdeburg, Germany
3Department Monitoring & Exploration Technologies, Helmholtz Centre for Environmental Research, Leipzig, Germany
4Department Aquatic Ecosystem Analysis, Helmholtz Centre for Environmental Research, Magdeburg, Germany
5Department Shelf Sea System Ecology, Alfred Wegener Institut, Helmholtz Zentrum für Polar- und Meeresforschung,
Helgoland, Germany
10 1.2 Traditional method for budgeting
35 Bottom-up approaches to quantifying riverine GHG emissions are typically based on GHG concentrations measured in a
restricted number of water samples. GHG fluxes between water and atmosphere are calculated from measured concentrations
and estimated gas transfer velocities (Raymond et al., 2013) multiplied by water surface area. Typical datasets contain weekly
to monthly concentration data from a small number of sites along a specific river. GHG emissions can also directly be measured by floating chambers. However, these measurements are laborious and prone to experimental artefacts if not carried out
40
carefully (Lorke et al., 2015). Water surface areas are typically estimated by using river width based on empirical relations
(Raymond et al., 2013) or for larger rivers using remote sensing data (Palmer and Ruhi, 2018). These approaches are suitable
to cover seasonal dynamics as well as large-scale spatial patterns along larger rivers. However, recent research has indicated
considerable short-term temporal and small-scale spatial variability of riverine GHG emissions which pose challenges for budgeting and upscaling. 45 budgeting and upscaling. 45 1.3 Short-term temporal variability The advent of reliable and affordable probes to continuously measure GHG concentrations revealed considerable diurnal
fluctuations of CO2 in rivers (Gómez-Gener et al., 2021). Because the balance between photosynthesis and respiration depends
on light, CO2 emissions are typically elevated in the night (Attermeyer et al., 2021). Thus, CO2 emission estimates only relying
on discrete water samples taken during daylight hours significantly under-estimate true emissions. While diurnal fluctuations
50
of CO2 are well documented, there is little knowledge concerning the short-term variability of CH4 (Stanley et al., 2016). The advent of reliable and affordable probes to continuously measure GHG concentrations revealed considerable diurnal
fluctuations of CO2 in rivers (Gómez-Gener et al., 2021). Because the balance between photosynthesis and respiration depends
on light, CO2 emissions are typically elevated in the night (Attermeyer et al., 2021). Thus, CO2 emission estimates only relying on light, CO2 emissions are typically elevated in the night (Attermeyer et al., 2021). Thus, CO2 emission estimates only relying
on discrete water samples taken during daylight hours significantly under-estimate true emissions. While diurnal fluctuations
50
of CO2 are well documented, there is little knowledge concerning the short-term variability of CH4 (Stanley et al., 2016). on discrete water samples taken during daylight hours significantly under-estimate true emissions. While diurnal fluctuations
50
of CO2 are well documented, there is little knowledge concerning the short-term variability of CH4 (Stanley et al., 2016). 1.1 Necessity of upscaling/quantification of GHG emissions from rivers Rivers are a globally relevant source of greenhouse gases (GHG) (Battin et al., 2023; Attermeyer et al., 2021; Raymond et al.,
2012). It is currently estimated that rivers globally emit about 2 Pt CO2 y-1 (Liu et al., 2022) and 30.5 ± 17.1Tg CH4 y-1
30 30 1 https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. (Rosentreter et al., 2021). However, these estimates suffer from considerable uncertainty. Aquatic GHG emissions actually
generate considerable uncertainty among global GHG assessments (IPCC, 2021). Reducing uncertainty in aquatic GHG
budgets is important in order to improve biogeochemical models and climate prediction(s). Uncertainties result from a general
lack of data as well as from the methods used for budgeting aquatic GHG emissions. 1.4 Small scale spatial variability While a single water sample might be representative of a certain specific reach in a small stream this is undoubtedly not the
case in larger rivers. Rivers contain various habitat types: these are either natural or those or with anthropogenic modifications. case in larger rivers. Rivers contain various habitat types: these are either natural or those or with anthropogenic modifications. Channelisation and disconnection of rivers from their floodplain decrease spatial heterogeneity (Wohl and Iskin, 2019) and
55
most likely also affect GHG emissions (Machado dos Santos Pinto et al., 2020). However, anthropogenic modifications not
only reduce habitat diversity. Several European rivers were modified by building groynes in the 19th century with the primary
goal of concentrating the water into the main river and to improve river flow and navigability (Pusch and Fischer, 2006). Consequently, the flow velocity is lower within the groyne fields leading to increased sedimentation (Kleinwächter et al.,
2017). Thus, groynes increase habitat diversity compared to straight, channelised rivers but decrease it compared to a natural
60 Channelisation and disconnection of rivers from their floodplain decrease spatial heterogeneity (Wohl and Iskin, 2019) and
55
most likely also affect GHG emissions (Machado dos Santos Pinto et al., 2020). However, anthropogenic modifications not
only reduce habitat diversity. Several European rivers were modified by building groynes in the 19th century with the primary
goal of concentrating the water into the main river and to improve river flow and navigability (Pusch and Fischer, 2006). Consequently, the flow velocity is lower within the groyne fields leading to increased sedimentation (Kleinwächter et al., 2017). Thus, groynes increase habitat diversity compared to straight, channelised rivers but decrease it compared to a natural
60 2 https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. shoreline. In the River Elbe, there are no groynes until German river km 120; however, they dominate the shore line
downstream from there (Fig. 1b). A recent study presents evidence that the still water areas between such groynes are a source
of CH4 resulting in lateral CH4 gradients (Bussmann et al., 2022). Ignoring these gradients significantly under-estimates total
CH4 emissions. shoreline. In the River Elbe, there are no groynes until German river km 120; however, they dominate the shore line
downstream from there (Fig. 1b). 1.5 Aim of this study We expected that small-scale spatial and diurnal variability would need to be considered for budgeting GHG emissions from
rivers. The ideal approach would thus be to perform high-frequency measurements at a large number of sites. However, this is
simply not possible and thus, there is a trade-off between frequency and spatial coverage of measurements. In this study, we
75
aim to answer the question “What is more important for budgeting/upscaling GHG emissions from rivers: small-scale spatial
or short-term temporal variability?”. We hypothesise that the answer to this question depends on the gas: We expected spatial
heterogeneity to be more relevant for CH4 emissions, while temporal variability is more relevant for CO2. To test this
hypothesis, we measured CH4 and CO2 emissions in different habitats within a typical reach of the German lowland river Elbe
over a four-day campaign. The study was designed to cover a typical low-discharge summer situation when both habitat
80
diversity and biological activity in the river were expected to be the highest. 1.4 Small scale spatial variability A recent study presents evidence that the still water areas between such groynes are a source
of CH4 resulting in lateral CH4 gradients (Bussmann et al., 2022). Ignoring these gradients significantly under-estimates total
CH4 emissions. A typical feature of rivers is their fluctuating discharge resulting in fluctuating water level. Thus, depending on discharge,
65
certain parts of rivers are temporarily drying up. It has been shown that these dry river areas emit disproportionally high
amounts of CO2 into the atmosphere (Gómez-Gener et al., 2015). Ignoring dry areas in river GHG budgets may lead to a
significant underestimation of GHG emissions (Marcé et al., 2019). Depending on their elevation, such dry river sediments
can be quite heterogeneous with respect either to substrate type (sand versus mud), or to the occurrence of temporary vegetation
(Bolpagni et al., 2019). Recent research indicates considerable spatial variability as well as temporal dynamics of dry river
70
GHG emissions (Mallast et al., 2020; Koschorreck et al., 2022). A typical feature of rivers is their fluctuating discharge resulting in fluctuating water level. Thus, depending on discharge,
65
certain parts of rivers are temporarily drying up. It has been shown that these dry river areas emit disproportionally high
amounts of CO2 into the atmosphere (Gómez-Gener et al., 2015). Ignoring dry areas in river GHG budgets may lead to a
significant underestimation of GHG emissions (Marcé et al., 2019). Depending on their elevation, such dry river sediments
can be quite heterogeneous with respect either to substrate type (sand versus mud), or to the occurrence of temporary vegetation 65 (Bolpagni et al., 2019). Recent research indicates considerable spatial variability as well as temporal dynamics of dry river
70
GHG emissions (Mallast et al., 2020; Koschorreck et al., 2022). (Bolpagni et al., 2019). Recent research indicates considerable spatial variability as well as temporal dynamics of dry river
70
GHG emissions (Mallast et al., 2020; Koschorreck et al., 2022). 2.1 Study Site Investigations were performed at an one km-long reach of the 8th order River Elbe at Tangermünde located in the middle part
of the river in Germany at river km 388 according to German kilometration (Figure 1a). The reach is typical for the middle
85
Elbe River which is characterised by groyne fields (Figure 1b) between km 120-580. Measurements were done in late summer
on 18-22 August 2022. Discharge varied between 189-197 m3 s-1 during that period which was below the mean low discharge
of 235 m3 s-1 (Figure 2). Investigations were performed at an one km-long reach of the 8th order River Elbe at Tangermünde located in the middle part of the river in Germany at river km 388 according to German kilometration (Figure 1a). The reach is typical for the middle
85
Elbe River which is characterised by groyne fields (Figure 1b) between km 120-580. Measurements were done in late summer
on 18-22 August 2022. Discharge varied between 189-197 m3 s-1 during that period which was below the mean low discharge
of 235 m3 s-1 (Figure 2). We separated the river's water surface into three distinct habitats: The middle of the river the sides of the river and the area We separated the river's water surface into three distinct habitats: The middle of the river, the sides of the river, and the area
between groynes (groyne fields). The groyne fields extend from the riverbank to a virtual line connecting the heads of the
90 between groynes (groyne fields). The groyne fields extend from the riverbank to a virtual line connecting the heads of the
90 between groynes (groyne fields). The groyne fields extend from the riverbank to a virtual line connecting the heads of the
90 3 https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. groynes. The side areas we defined as extending from the outer boundary of the groyne fields 15 m into the river. The studied
reach had ten groynes at both banks, extending up to 60 m into the river. The distance between the groynes was 80 ± 10 m. The area between the groynes was partly dry. These dry areas featured three typical habitats: muddy areas and sandy beaches
without and with terrestrial vegetation (Figure 1c, Figure S1). 2.1 Study Site 95
Figure 1: Location of the investigation site Tangermünde (red dot) within Germany at River Elbe at river km 388 according to
German kilometration (a). Top view of the sampling reach with groyne fields (Source orthophoto: Google Earth, GeoBasis-DE/BKG,
date of recording 06/08/2020), flow direction to North-east, study area marked by red lines (b). Detailed view of the study area with
indication of habitat types (c). The groyne field is divided into an aquatic habitat and the terrestrial habitat with partially dry fallen
sediments, which are divided into sandy areas (location of soil flux chamber C1), muddy areas (chamber C2) and vegetated areas
100
(chamber C3). (Source orthophoto: Google Earth, GeoBasis-DE/BKG, recording date 06/08/2020, © Google Earth, see also Fig. S1). 95 Figure 1: Location of the investigation site Tangermünde (red dot) within Germany at River Elbe at river km 388 according to
German kilometration (a). Top view of the sampling reach with groyne fields (Source orthophoto: Google Earth, GeoBasis-DE/BKG,
date of recording 06/08/2020), flow direction to North-east, study area marked by red lines (b). Detailed view of the study area with
indication of habitat types (c). The groyne field is divided into an aquatic habitat and the terrestrial habitat with partially dry fallen
sediments, which are divided into sandy areas (location of soil flux chamber C1), muddy areas (chamber C2) and vegetated areas
100
(chamber C3). (Source orthophoto: Google Earth, GeoBasis-DE/BKG, recording date 06/08/2020, © Google Earth, see also Fig. S1). 4 https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. Figure 2: Discharge of River Elbe at gauge Tangermünde during 2022 (German Federal Waterways and Shipping Administration). The blue area marks the range between mean low discharge and mean discharge, the red area marks the period of the sampling
campaign. 5 Figure 2: Discharge of River Elbe at gauge Tangermünde during 2022 (German Federal Waterways and Shipping Administration). The blue area marks the range between mean low discharge and mean discharge, the red area marks the period of the sampling
campaign. 2.2.1 Flow velocity and water depth measurement We mapped basic hydrographic parameters with the RV Albis meandering between the western and
eastern groyne heads on 19 August 2022. Due to the size of the ship, it was not possible to enter near-shore areas and groyne
fields From 19 22 August 2022
the RV Albis was anchored at the pier in Tangermünde and measured the hydrographic
120 (GPS) position data were collected using a GPS tracker (Garmin International) with a frequency of 1s. 115
Basic hydrographic parameters (temperature, conductivity, oxygen, pH, turbidity and chlorophyll) were determined with a
pocketferrybox (4Hjena, Kiel, Germany) and a multiparameter probe (Exo2, YSI). The water supply for both sensors was the
moon pool of the Albis. We mapped basic hydrographic parameters with the RV Albis meandering between the western and
eastern groyne heads on 19 August 2022. Due to the size of the ship, it was not possible to enter near-shore areas and groyne
fields. From 19-22 August 2022, the RV Albis was anchored at the pier in Tangermünde and measured the hydrographic
120
parameters at the same position, continuously for 63 hours. moon pool of the Albis. We mapped basic hydrographic parameters with the RV Albis meandering between the western and
eastern groyne heads on 19 August 2022. Due to the size of the ship, it was not possible to enter near-shore areas and groyne
fields. From 19-22 August 2022, the RV Albis was anchored at the pier in Tangermünde and measured the hydrographic
120
parameters at the same position, continuously for 63 hours. fields. From 19-22 August 2022, the RV Albis was anchored at the pier in Tangermünde and measured the hydrographic
120
parameters at the same position, continuously for 63 hours. fields. From 19-22 August 2022, the RV Albis was anchored at the pier in Tangermünde and measured the hydrographic
120
parameters at the same position, continuously for 63 hours. fields. From 19-22 August 2022, the RV Albis was anchored at the pier in Tangermünde and measured the hydrographic
120
parameters at the same position, continuously for 63 hours. 2.2.1 Flow velocity and water depth measurement Flow velocity profile measurements were conducted at the study site at Tangermünde on 19 August 2022. We deployed a
Teledyne RD Instruments (TRDI) four-beam 1200 kHz Rio Grande ADCPTM from an inflatable boat. The vertical resolution
of water velocities was 0.25 m (25 cm bins), and the sampling frequency was approximately 2 Hz. In total 8 transects of flow
110
velocity and water depth where measured using 4-5 replicates at each transect (Figure S2 in the Supplement). The minimum
and maximum measured flow velocity were 0.007 and 1.05 ms-1, and the minimum and maximum water depths were 0.45 and
3.1 m respectively. Depth-averaged velocities (U) were calculated from the measured portion of the water column, neglecting y p
y
g
Teledyne RD Instruments (TRDI) four-beam 1200 kHz Rio Grande ADCPTM from an infl Teledyne RD Instruments (TRDI) four-beam 1200 kHz Rio Grande ADCPTM from an inflatable boat. The vertical resolution
of water velocities was 0.25 m (25 cm bins), and the sampling frequency was approximately 2 Hz. In total 8 transects of flow
110
velocity and water depth where measured using 4-5 replicates at each transect (Figure S2 in the Supplement). The minimum
and maximum measured flow velocity were 0.007 and 1.05 ms-1, and the minimum and maximum water depths were 0.45 and
3.1 m respectively. Depth-averaged velocities (U) were calculated from the measured portion of the water column, neglecting 5 https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. the unmeasured upper and lower portion of the water column (see Figure S2 in the Supplement). Global positioning system the unmeasured upper and lower portion of the water column (see Figure S2 in the Supplement). Global positioning system
(GPS) position data were collected using a GPS tracker (Garmin International) with a frequency of 1s. 115 the unmeasured upper and lower portion of the water column (see Figure S2 in the Suppleme (GPS) position data were collected using a GPS tracker (Garmin International) with a frequency of 1s. 115
Basic hydrographic parameters (temperature, conductivity, oxygen, pH, turbidity and chlorophyll) were determined with a
pocketferrybox (4Hjena, Kiel, Germany) and a multiparameter probe (Exo2, YSI). The water supply for both sensors was the
moon pool of the Albis. 2.2.2 Aquatic GHG measurements To assess the spatial pattern of aquatic GHG emissions, aquatic GHG fluxes were measured from an inflatable boat using a
floating chamber connected to a portable FTIR analyser (GASMET) as explained in Lorke et al. (2015). For each measurement
the chamber was deployed from a drifting boat for 2-5 minutes. Fluxes were calculated from the linear change of CO2 and CH4
125 To assess the spatial pattern of aquatic GHG emissions, aquatic GHG fluxes were measured
floating chamber connected to a portable FTIR analyser (GASMET) as explained in Lorke et al To assess the spatial pattern of aquatic GHG emissions, aquatic GHG fluxes were measured from an inflatable boat using a
floating chamber connected to a portable FTIR analyser (GASMET) as explained in Lorke et al. (2015). For each measurement floating chamber connected to a portable FTIR analyser (GASMET) as explained in Lorke et al. (2015). For each measurement
the chamber was deployed from a drifting boat for 2-5 minutes. Fluxes were calculated from the linear change of CO2 and CH4
125
mixing ratios in the chamber. A water sample for later gas chromatograph (GC) analysis was taken during each flux
measurement. Surface water samples of 30 mL were taken with 60 mL syringes, and 30 ml of ambient air was added to the
syringes, which was then vigorously shaken for one minute. The equilibrated headspace was then transferred to pre-evacuated
exetainers and the equilibration temperature was measured in the remaining water sample. Dissolved methane concentrations were mapped with a dissolved gas extraction unit and a laser-based analytical Greenhouse
130
Gas Analyzer (GGA; both Los Gatos Research, United States) on an inflatable boat. The degassing unit withdrew water from
the water basin or directly from surface water at 1.2 L min 1-1. Methane was extracted from the water via a hydrophobic
membrane and hydrocarbon-free carrier gas was on the other side of the membrane (nitrogen, at 0.5 L min-1). The carrier gas
with the extracted CH4 was then directed to the inlet of the gas analyser. The time offset between the water intake and stable Dissolved methane concentrations were mapped with a dissolved gas extraction unit and a laser-based analytical Greenhouse
130
Gas Analyzer (GGA; both Los Gatos Research, United States) on an inflatable boat. The degassing unit withdrew water from
the water basin or directly from surface water at 1.2 L min 1-1. 2.2.2 Aquatic GHG measurements Methane was extracted from the water via a hydrophobic
membrane and hydrocarbon-free carrier gas was on the other side of the membrane (nitrogen, at 0.5 L min-1). The carrier gas
with the extracted CH4 was then directed to the inlet of the gas analyser. The time offset between the water intake and stable recording at the GGA was determined beforehand in the laboratory. To convert the relative concentrations (ppm) given by the
135
GGA to absolute concentrations (nmol L-1), discrete water samples were obtained at least every hour. The CH4 concentration
in these bottles was determined using the headspace method and gas chromatographic analysis. Based on the obtained values,
the conversion factor (ppm to nmol L-1) of 84.88 was determined. For high-spatial resolution measurements the instruments were set-up in a small rubber boat with a 5-L nitrogen tank and a car recording at the GGA was determined beforehand in the laboratory. To convert the relative concentrations (ppm) given by the
135
GGA to absolute concentrations (nmol L-1), discrete water samples were obtained at least every hour. The CH4 concentration
in these bottles was determined using the headspace method and gas chromatographic analysis. Based on the obtained values,
the conversion factor (ppm to nmol L-1) of 84.88 was determined. For high-spatial resolution measurements the instruments were set-up in a small rubber boat with a 5-L nitrogen tank and a car recording at the GGA was determined beforehand in the laboratory. To convert the relative concentrations (ppm) given by the
135
GGA to absolute concentrations (nmol L-1), discrete water samples were obtained at least every hour. The CH4 concentration
in these bottles was determined using the headspace method and gas chromatographic analysis. Based on the obtained values,
the conversion factor (ppm to nmol L-1) of 84.88 was determined. For high spatial resolution measurements the instruments were set up in a small rubber boat with a 5 L nitrogen tank and a car For high-spatial resolution measurements the instruments were set-up in a small rubber boat with a 5-L nitrogen tank and a car
battery. The water inlet of the degasser was fixed to a bar and submerged to approx. 20 cm water depth. We entered each
140
groyne field from the north and kept the boat at the groyne heads for approximately 2 min. (against the current) and then
entered the following groyne field as far as possible (Figure 5). battery. 2.2.2 Aquatic GHG measurements Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. CH4 mixing ratio (ppm) given by the Contros Sensor was corrected to absolute concentrations (µmol L-1) using the CH4
concentration in water samples taken during the measuring interval and analysed with gas chromatography. 2.3 Terrestrial measurements Based on our own long-term tests with the system,
160
soil flux detection limits were determined for CO2 of ±0.36 mmol m-2 h-1 (corresponding to a change of 2 ppm CO2 during a
closure time period of 2 minutes) and for CH4 of ±0.072 µmol m-2 h-1 (corresponding to a change of 0.6 ppb CH4 during a
closure time period of 2 minutes). To cover the temporal dynamics of terrestrial CO2 emissions, three opaque automatic chambers (CFLUX-1 Automated Soil CO2 Flux System, PP systems, Amesbury, Massachusetts, USA) were installed at a sandy site, a muddy site, and a sandy site
165
with herbaceous vegetation (Figure 1 d). The chambers measured CO2 fluxes once every hour. Each flux measurement lasted
for 5 min, and the chambers were open for 55 min between flux measurements. CO2 fluxes were calculated from the linear
increase of CO2. The detection limit for terrestrial CO2 fluxes was 0.08 mmol m-2 h-1. Each chamber was equipped with a soil
moisture and temperature probe (Stevens HydraProbe, Stevens Water Monitoring Systems, Portland, Oregon, USA). CO2 Flux System, PP systems, Amesbury, Massachusetts, USA) were installed at a sandy site, a muddy site, and a sandy site
165
with herbaceous vegetation (Figure 1 d). The chambers measured CO2 fluxes once every hour. Each flux measurement lasted
for 5 min, and the chambers were open for 55 min between flux measurements. CO2 fluxes were calculated from the linear
increase of CO2. The detection limit for terrestrial CO2 fluxes was 0.08 mmol m-2 h-1. Each chamber was equipped with a soil
moisture and temperature probe (Stevens HydraProbe, Stevens Water Monitoring Systems, Portland, Oregon, USA). Light intensity was measured at Magdeburg (50 km from Tangermünde) as PAR [µmol m-2 s-1] using a LI-190R Quantum
170
Sensor (Licor, Lincoln, U.S.A.). Light intensity was measured at Magdeburg (50 km from Tangermünde) as PAR [µmol m-2 s-1] using a LI-190R Quantum
170
Sensor (Licor, Lincoln, U.S.A.). Light intensity was measured at Magdeburg (50 km from Tangermünde) as PAR [µmol m-2 s-1] using a LI-190R Quantum
170
Sensor (Licor, Lincoln, U.S.A.). 2.2.2 Aquatic GHG measurements The water inlet of the degasser was fixed to a bar and submerged to approx. 20 cm water depth. We entered each
140
groyne field from the north and kept the boat at the groyne heads for approximately 2 min. (against the current) and then
entered the following groyne field as far as possible (Figure 5). For continuous measurements of dissolved CO2 and CH4, an optical AMT-Sensor (Rostock, Germany) and a Contros-Sensor battery. The water inlet of the degasser was fixed to a bar and submerged to approx. 20 cm water depth. We entered each
140
groyne field from the north and kept the boat at the groyne heads for approximately 2 min. (against the current) and then
entered the following groyne field as far as possible (Figure 5). For continuous measurements of dissolved CO2 and CH4, an optical AMT-Sensor (Rostock, Germany) and a Contros-Sensor
(4H J
J
G
)
d
l
d i th
l
f th Albi
Th CO
id d d t
hi h battery. The water inlet of the degasser was fixed to a bar and submerged to approx. 20 cm water depth. We entered each
140
groyne field from the north and kept the boat at the groyne heads for approximately 2 min. (against the current) and then
entered the following groyne field as far as possible (Figure 5). For continuous measurements of dissolved CO2 and CH4, an optical AMT-Sensor (Rostock, Germany) and a Contros-Sensor
(4H-Jena, Jena, Germany) were deployed in the moon pool of the Albis. The CO2 sensor provided data as ppm which were
converted to concentrations (µmol L-1) according to its solubility at the respective temperature (UNESCO/IHA, 2010). The
145 entered the following groyne field as far as possible (Figure 5). For continuous measurements of dissolved CO2 and CH4, an optical AMT-Sensor (Rostock, Germany) and a Contros-Sensor
(4H-Jena, Jena, Germany) were deployed in the moon pool of the Albis. The CO2 sensor provided data as ppm which were For continuous measurements of dissolved CO2 and CH4, an optical AMT-Sensor (Rostock, Germany) and a Contros-Sensor
(4H-Jena, Jena, Germany) were deployed in the moon pool of the Albis. The CO2 sensor provided data as ppm which were
converted to concentrations (µmol L-1) according to its solubility at the respective temperature (UNESCO/IHA, 2010). The
145 converted to concentrations (µmol L-1) according to its solubility at the respective temperature (UNESCO/IHA, 2010). The
145 6 https://doi.org/10.5194/bg-2023-176
Preprint. 2.3 Terrestrial measurements The mixing ratio of CO2 in the atmosphere and other meteorological parameters was continuously measured by a sensebox. The sensebox system is a toolkit developed in the framework of Citizen Science projects for environmental data collection
150
funded by the German Federal Ministry of Education and Research. It consists of an open-source microcontroller unit which
can deploy various environmental sensors (Bartoscheck et al., 2019). The sensebox was equipped with a GPS-sensor, an
environmental sensor measuring air temperature, relative humidity and air pressure, and a CO2 - sensor determining the air
CO2 mixing ratio. The sensebox was installed at the RV Albis and measured the atmospheric conditions every 5 minutes. The spatial variability of CO2 and CH4 fluxes at terrestrial sediments was measured with the laser-based trace gas analyser LI-
155
7810 (LI-COR Biosciences, USA) in combination with a closed chamber (LI-COR smart chamber). Soil gas fluxes were
calculated from the temporal gas concentration change taking into consideration the chamber volume and the surface area of
the soil area covered by the chamber. For each sampling point the change in gas concentration in the closed chamber was
determined after a purging period in a 1-second sampling interval during the 2-minute observation time. Fluxes were calculated The spatial variability of CO2 and CH4 fluxes at terrestrial sediments was measured with the laser-based trace gas analyser LI-
155
7810 (LI-COR Biosciences, USA) in combination with a closed chamber (LI-COR smart chamber). Soil gas fluxes were
calculated from the temporal gas concentration change taking into consideration the chamber volume and the surface area of
the soil area covered by the chamber. For each sampling point the change in gas concentration in the closed chamber was
determined after a purging period in a 1-second sampling interval during the 2-minute observation time. Fluxes were calculated using the linear fitting approach of the SoilFluxPro Software (LI-COR). Based on our own long-term tests with the system,
160
soil flux detection limits were determined for CO2 of ±0.36 mmol m-2 h-1 (corresponding to a change of 2 ppm CO2 during a
closure time period of 2 minutes) and for CH4 of ±0.072 µmol m-2 h-1 (corresponding to a change of 0.6 ppb CH4 during a
closure time period of 2 minutes). To cover the temporal dynamics of terrestrial CO2 emissions, three opaque automatic chambers (CFLUX-1 Automated Soil using the linear fitting approach of the SoilFluxPro Software (LI-COR). 2.5 Calculations and Statistics Gas transfer coefficients were calculated from CH4 fluxes measured by the floating chamber Gas transfer coefficients were calculated from CH4 fluxes measured by the floating chambers divided by the difference between
actual and equilibrium CH4 concentration. Equilibrium CH4 concentrations were calculated from the mean measured
180
atmospheric CH4 mixing ratio (2.5 ppm) using temperature-dependent Henry coefficients from Sander (2015); (Koschorreck
et al., 2021). The so-determined kCH4 was converted to k600 and kCO2 using Schmidt numbers according to UNESCO/IHA
(2010). We did not use CO2 data for k600 calculations because the CO2 concentration was close to equilibrium resulting in large
uncertainties in the calculation of k600. 180 Probe measurements of CO2 and CH4 concentrations were converted to fluxes using the measured gas transfer velocity of k600
185
= 5.5 m d-1 (Table 1). k600 was converted to kCO2 and kCH4 as explained in Striegl et al. (2012) . Emissions from different habitat types were compared by pairwise Wilcoxon tests. Fluxes during the day were compared to
fluxes during night using Wilcoxon rank sum tests. For statistical analysis, all high-frequency data were transformed to hourly
data by calculating the mean values of data available between 30 minutes before and 30 minutes after the full hour. Time series Probe measurements of CO2 and CH4 concentrations were converted to fluxes using the measured gas transfer velocity of k600
185
= 5.5 m d-1 (Table 1). k600 was converted to kCO2 and kCH4 as explained in Striegl et al. (2012) . data were log transformed after checking for normality by using Shapiro-Wilk tests. Significance of linear correlation of log
190
transformed fluxes with drivers was checked by F-tests (p<0.05). Mixed linear models to explain GHG fluxes from
combinations of predictors were compared based on their AIC. All statistical analyses were done with R (R-Core-Team, 2016). 2.4 Laboratory Analyses CO2 and CH4 concentrations in gas samples were measured with a gas chromatograph (GC) (SRI 8610C, SRI Instruments
Europe, Bad Honnef, Germany). The GC was equipped with a flame ionisation detector and a methanizer which allowed for
simultaneous measurement of CO2 and CH4 with an uncertainty of < 5 %. Dissolved gas concentrations were calculated using
175 175 7 https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. temperature-dependent Henry coefficients (UNESCO/IHA, 2010). CO2 concentrations were corrected for alkalinity as
described in Koschorreck et al. (2021). temperature-dependent Henry coefficients (UNESCO/IHA, 2010). CO2 concentrations were corrected for alkalinity as
described in Koschorreck et al. (2021). gradient has been attributed to the salty inflow of river Saale 97 km upstream of our sampling site (Weigold and Baborowski,
2009). It indicates limited lateral mixing of river water even over a large distance. gradient has been attributed to the salty inflow of river Saale 97 km upstream of our sampling site (Weigold and Baborowski,
2009). It indicates limited lateral mixing of river water even over a large distance. 205 3.2 Spatial variability GHG concentrations and emissions differed between habitats and also between the two gases CO2 and CH4 (Table 1). The
water was over-saturated both with CO2 and CH4, resulting in positive fluxes (= emission to the atmosphere). The flux of CO2
from the water surface was two orders of magnitude higher than the CH4 flux. Dissolved CO2 concentrations were highest in
210
the middle of the river and decreased towards the side. An opposite pattern was observed for CH4 which was higher at the side
and within the groyne fields than in the middle of the river (Figure 3). Since the gas transfer velocity k600 was twice as high
at the sites compared to the middle of the river, CH4 emissions were significantly higher at the side and in the groyne field
compared to the middle of the river. Since higher CO2 concentrations in the middle of the river were compensated by lower
k600 values, emissions of CO2 did not differ significantly between aquatic habitats (Figure 4). 215 GHG concentrations and emissions differed between habitats and also between the two gases CO2 and CH4 (Table 1). The
water was over-saturated both with CO2 and CH4, resulting in positive fluxes (= emission to the atmosphere). The flux of CO2 from the water surface was two orders of magnitude higher than the CH4 flux. Dissolved CO2 concentrations were highest in
210
the middle of the river and decreased towards the side. An opposite pattern was observed for CH4 which was higher at the side
and within the groyne fields than in the middle of the river (Figure 3). Since the gas transfer velocity k600 was twice as high
at the sites compared to the middle of the river, CH4 emissions were significantly higher at the side and in the groyne field
compared to the middle of the river. Since higher CO2 concentrations in the middle of the river were compensated by lower k600 values, emissions of CO2 did not differ significantly between aquatic habitats (Figure 4). 215
The dry habitats had higher CO2 emissions but lower CH4 emissions compared to the aquatic sites. Dry CO2 emissions showed
a large variability within habitats and were highest at vegetated sites and very low at the sandy sites (Figure 4). 3.1 Hydrodynamic and climatic conditions Our study was conducted during a typical summer low-water situation. At the gauge level of 134 cm (on 6 August 2022, when
195
the orthophoto in Figure 1 was taken), 9 % of the study area was not covered by water (Table 1). Flow velocity in the river
was relatively uniform around 0.65 m s-1 while within the groyne fields flow velocity was significantly lower (Table 1). Water
was flowing rather smoothly without larger waves, but we observed more turbulent conditions downstream of groyne heads. The mean water depth was 1.82 m with the most shallow but also the deepest parts in the groyne fields (maximum depth 3.1 m). The water level rose by 10 cm over the 3 days of our study. Weather conditions were rather constant during our study
200
period, predominantly sunny, with 5.6 mm of rainfall during the first night (19.8. until 6 am). Air temperature fluctuated
between 12.4° and 32.2° and low wind speeds of 0.3 ± 0.6 m s-1 were recorded. Water temperatures were evenly distributed
(23.3 ± 0.04°C, measured between 19.08.2022 14:00 and 22.08.2022 08:00). Electrical conductivity of the water at the western
shore and within the western groyne fields was about 200 µS cm-1 higher than on the eastern shore (Fig. S3). This conductivity 8 https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. gradient has been attributed to the salty inflow of river Saale 97 km upstream of our sampling site (Weigold and Baborowski,
2009). It indicates limited lateral mixing of river water even over a large distance. 3.2 Spatial variability Dry CH4
emissions, in contrast, were more than two orders of magnitude lower, in some cases even slightly negative (Table 1), and
only small differences between habitats were observed. The dry habitats had higher CO2 emissions but lower CH4 emissions compared to the aquatic sites. Dry CO2 emissions showed
a large variability within habitats and were highest at vegetated sites and very low at the sandy sites (Figure 4). Dry CH4
emissions, in contrast, were more than two orders of magnitude lower, in some cases even slightly negative (Table 1), and
only small differences between habitats were observed. 220
Figure 3: Concentrations of dissolved CH4 in the water measured continuously with a mobile gas extraction unit connected to a
GHG analyser. Measurements were carried out from an inflatable boat on 21.8.2022 between 16:18 and 16:51. 220 Figure 3: Concentrations of dissolved CH4 in the water measured continuously with a mobile gas extraction unit connected to a
GHG analyser. Measurements were carried out from an inflatable boat on 21.8.2022 between 16:18 and 16:51. 9 https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. Table 1: GHG emissions and their regulating factors in different habitat types. Emission data from floating chamber
measurements, CH4 concentration measured by a membrane equilibrator connected to a portable GHG analyser (data from
5
Figure 3), CO2 concentration analysed by GC, velocity measured by ADCP. Areas were manually extracted from a recent
Google earth image (Figure 1). all data: median (range). 3.2 Spatial variability CH4 fluxes are given in μmol m-2 h-1 and CO2 fluxes in mmol m-2 h-
1 aquatic
terrestrial
middle
side
groyne fields
sandy
muddy
vegetated
CH4
emissions
[μmol m-2 h-1]
12.2
(6-20.7)
24.9
(6-73)
39
(12.6-52.6)
0.2
(-0.06 - 0.45)
0.1
(-0.8 - 2.5)
-0.1
(-2.8 - 0.4)
CO2
emissions
[mmol m-2 h-1]
2.2
(1.86-2.21)
1.5
(1.07-3.14)
0.8
(0.21-2.3)
0.6
(0.4 - 1.3)
1.3
(-1.5 - 3.9)
2.8
(0.2 - 13.9)
CH4
concentration
[µmol L-1]
0.12
(0.11 - 0.16)
0.18
(0.17 - 0.32)
0.18
(0.17 -0.21)
-
-
-
CO2
concentration
[µmol L-1]
29.3
(28.4-29.6)
23.4
(15.4-24.3)
15.7
(15.4-28.8)
-
-
-
k600 [m d-1]
2.8
(1.7-5.5)
5.7
(2.3-10.8)
4.9
(1.5-6)
-
-
-
velocity [m s-1]
0.79
(0.72-0.81)
0.65
(0.22-0.84)
0.22
(0.09-0.45)
-
-
-
total area [m2]
125,000
25,000
42,000
7,700
4,800
6,500
total CH4 emissions
[mol h-1]
1.5
(0.7 - 2.6)
0.6
(0.1 - 1.8)
1.6
(0.5 - 2.2)
0.0015
(-0.0005 -
0.0035)
0.0005
(-0.0038 - 0.012)
-0.00065
(-0.018 -
0.0026)
total CO2 emissions
[mol h-1]
275
(232 - 276)
37.5
(26.7 - 78.5)
33.6
(8.8 - 96.6)
4.6
(3.1 - 10.0)
6.2
(-7.2 - 18.7)
18.2
(1.3 - 90.3) 37.5
(26.7 - 78.5) 10 230
Figure 4: Boxplots comparing CO2 flux (a) and CH4 flux (b) in different habitat types. Numbers within the plots indicate p-values
of pairwise comparisons using Wilcoxon tests. https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. 30 Figure 4: Boxplots comparing CO2 flux (a) and CH4 flux (b) in different habitat types. Numbers within the plots indicate p-values
of pairwise comparisons using Wilcoxon tests. 3.3 Temporal variability Continuous measurements of aquatic GHG concentrations (Figure S4) and water physicochemical variables were carried out Continuous measurements of aquatic GHG concentrations (Figure S4) and water physi Continuous measurements of aquatic GHG concentrations (Figure S4) and water physicochemical variables were carried out
over a period of 2.5 days at the side of the river. Water temperature (median 22.6; range 21.7 - 23.7°C), pH (median 8.0; range
35
7 3
8 3) and oxygen (median 97 9; 87 5
117 8 % saturation) showed a clear daily pattern with lowest values in the early Continuous measurements of aquatic GHG concentrations (Figure S4) and water physicochemical variables were carried out
over a period of 2.5 days at the side of the river. Water temperature (median 22.6; range 21.7 - 23.7°C), pH (median 8.0; range
235
7.3 - 8.3) and oxygen (median 97.9; 87.5 - 117.8 % saturation) showed a clear daily pattern, with lowest values in the early
morning (5:00 - 6:00), while conductivity was rather constant (median 1260; range 1146 - 1381 µS/cm; Supp. Figure S5). Aquatic CO2 concentrations showed a clear diurnal cycle with rising concentrations during the night and decreasing
concentrations during the day. The diurnal amplitude of the resulting flux spanned about 5 mmol m2 h-1 with maximum fluxes over a period of 2.5 days at the side of the river. Water temperature (median 22.6; range 21.7 - 23.7°C), pH (median 8.0; range
235
7.3 - 8.3) and oxygen (median 97.9; 87.5 - 117.8 % saturation) showed a clear daily pattern, with lowest values in the early
morning (5:00 - 6:00), while conductivity was rather constant (median 1260; range 1146 - 1381 µS/cm; Supp. Figure S5). Aquatic CO2 concentrations showed a clear diurnal cycle with rising concentrations during the night and decreasing
concentrations during the day. The diurnal amplitude of the resulting flux spanned about 5 mmol m2 h-1 with maximum fluxes around 7:00 and minimum fluxes around 17:00 (blue line in Figure 5a). In contrast to CO2, the CH4 concentration and the flux
240
did not change with time (Figure 5b). At the dry sites diurnal pattern of CO2 emissions are apparent, but these patterns differed between habitat type. At the vegetated
site a pattern similar to the aquatic site was observed, but phase shifted. The highest fluxes were found around noon, while the
minimum was at 4:00 before sunrise (green line in Figure 5a). 3.3 Temporal variability The mud site did not show such a sinus-like pattern but rather a two-state pattern: The site switched from constant emissions during the night to constant CO2 uptake during the day (red line
245
in Figure 5a). No diurnal variability of CO2 emissions was observed at the sandy site (yellow line in Figure 5a). The CH4 flux
data from the dry sites do not allow visualisation of diurnal variability, but the data (dots in Figure 5b) at least do not show
any temporal trend during the day. CO2 emissions from the mud and the vegetated site showed a decreasing trend during our study. At the muddy site this trend CO2 emissions from the mud and the vegetated site showed a decreasing trend during our study. At the muddy site this trend
was associated with increasing sediment moisture (Figure 5c), which was obviously caused by the rising water level of the
250 was associated with increasing sediment moisture (Figure 5c), which was obviously caused by the rising water level of the
250 11 river (Figure 5d) - on 21.8. the flux chamber was only 1 m from the water line. At the other dry sites no trend of sediment
moisture was visible, but sediment temperature during the day tended to increase through the study (Figure 5c). river (Figure 5d) - on 21.8. the flux chamber was only 1 m from the water line. At the other dry sites no trend of sediment
moisture was visible, but sediment temperature during the day tended to increase through the study (Figure 5c). river (Figure 5d) - on 21.8. the flux chamber was only 1 m from the water line. At the other dry sites no trend of sediment
moisture was visible, but sediment temperature during the day tended to increase through the study (Figure 5c). Figure 5: Time series of CO2 flux (a), CH4 flux (b), sediment and water temperature (c), and sediment moisture and water level (d). All data with hourly resolution. Black line in c) shows light measured as PAR [µmol m-2 s-1/2]. Grey-shaded areas indicate night. 5
As a result of these diurnal patterns, fluxes were significantly higher during the day than during the night at the sandy and
vegetated site, while at the muddy site fluxes were higher at night than during daylight (Table 2). At the water site, medians
did not differ significantly between day and night. 3.3 Temporal variability Furthermore, the temperature of the water was relatively constant – in contrast to the dry sites where diurnal temperature
amplitudes up to 20° were observed (Figure 5c). If we correlate the CO2 flux with temperature or light for each habitat type
separately, we get a significant positive correlation at the sandy and vegetated site, while the correlation was negative at the
muddy site (Figure S7). A positive correlation with light was observed at the vegetated site, while the correlation was negative
265
at the muddy site. To consider site-specific correlations, we added site as a random factor to our statistical model. A mixed linear model with
(log-transformed) temperature, light, water level, and sediment moisture as fixed factors and site as random factor explained
82 % of the variability. The best-simplified model (based on AIC) contained water level and sediment moisture as fixed factors
and had a conditional R2 of 0.82. 270 including all data was not significant (F-test, p>0.05), which is consistent with the observed different diurnal pattern at different
sites. Furthermore, the temperature of the water was relatively constant – in contrast to the dry sites where diurnal temperature
amplitudes up to 20° were observed (Figure 5c). If we correlate the CO2 flux with temperature or light for each habitat type
separately, we get a significant positive correlation at the sandy and vegetated site, while the correlation was negative at the
muddy site (Figure S7). A positive correlation with light was observed at the vegetated site, while the correlation was negative
65
at the muddy site. including all data was not significant (F-test, p>0.05), which is consistent with the observed different diurnal pattern at different
sites. Furthermore, the temperature of the water was relatively constant – in contrast to the dry sites where diurnal temperature
amplitudes up to 20° were observed (Figure 5c). If we correlate the CO2 flux with temperature or light for each habitat type
separately, we get a significant positive correlation at the sandy and vegetated site, while the correlation was negative at the
muddy site (Figure S7). A positive correlation with light was observed at the vegetated site, while the correlation was negative
265
at the muddy site. To consider site-specific correlations, we added site as a random factor to our statistical model. 3.3 Temporal variability A mixed linear model with
(log-transformed) temperature, light, water level, and sediment moisture as fixed factors and site as random factor explained
82 % of the variability. The best-simplified model (based on AIC) contained water level and sediment moisture as fixed factors
and had a conditional R2 of 0.82. 270 Table 2: Median (range) of temporal GHG fluxes at different sites (data from Figure 7). Day and night separated by sunrise/sunset
Day and night data were significantly different for the dry sites but not for the water (Wilcox test). habitat
all data
day
night
CO2 flux
mud
2.3 (-2.34 - 6.41)
0.07 (-2.34 - 6.16)
2.97 (1.3 - 6.41)
sand
1.19 (-0.22 - 2.45)
1.37 (0.43 - 2.45)
0.79 (-0.22 - 1.91)
vegetation
7.06 (3.24 - 11.84)
8.17 (4.21 - 11.84)
6.01 (3.24 - 8.75)
water
3.32 (0.80 - 5.96)
2.94 (0.80 - 5.96)
3.53 (1.26 - 5.77)
CH4 flux
water
0.052 (0.05 - 0.055)
0.053 (0.05 - 0.054)
0.053 (0.052 - 0.055) Table 2: Median (range) of temporal GHG fluxes at different sites (data from Figure 7). Day and night separated by sunrise/sunset. Day and night data were significantly different for the dry sites but not for the water (Wilcox test). Table 2: Median (range) of temporal GHG fluxes at different sites (data from Figure 7). Day and nigh
Day and night data were significantly different for the dry sites but not for the water (Wilcox test). 3.3 Temporal variability Figure 5: Time series of CO2 flux (a), CH4 flux (b), sediment and water temperature (c), and sediment moisture and water level (d). All data with hourly resolution. Black line in c) shows light measured as PAR [µmol m-2 s-1/2]. Grey-shaded areas indicate night. 255 Figure 5: Time series of CO2 flux (a), CH4 flux (b), sediment and water temperature (c), and sediment moisture and water level (d). All data with hourly resolution. Black line in c) shows light measured as PAR [µmol m-2 s-1/2]. Grey-shaded areas indicate night. 255 Figure 5: Time series of CO2 flux (a), CH4 flux (b), sediment and water temperature (c), and sediment moisture and water level (d). All data with hourly resolution. Black line in c) shows light measured as PAR [µmol m-2 s-1/2]. Grey-shaded areas indicate night. 255
As a result of these diurnal patterns, fluxes were significantly higher during the day than during the night at the sandy and
vegetated site, while at the muddy site fluxes were higher at night than during daylight (Table 2). At the water site, medians
did not differ significantly between day and night. W
i d
l
h
d
CO fl
i h
d
i l d i
(Fi
S6) Th
CO fl
kl
i
l As a result of these diurnal patterns, fluxes were significantly higher during the day than during the night at the sandy and
vegetated site, while at the muddy site fluxes were higher at night than during daylight (Table 2). At the water site, medians
did not differ significantly between day and night. We tried to correlate the dry CO2 flux with measured potential drivers (Figure S6). The CO2 flux was weakly negatively
correlated with sediment moisture and water level. Correlation with temperature or light (which were highly auto-correlated)
260 We tried to correlate the dry CO2 flux with measured potential drivers (Figure S6). The CO2 flux was weakly negatively
correlated with sediment moisture and water level. Correlation with temperature or light (which were highly auto-correlated)
260 correlated with sediment moisture and water level. Correlation with temperature or ligh
260 correlated with sediment moisture and water level. Correlation with temperature or light (which were highly auto-correlated)
260 12 https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. including all data was not significant (F-test, p>0.05), which is consistent with the observed different diurnal pattern at different
sites. 3.4 Comparison of spatial and temporal variability
275 Figure 6: Coefficients of variation of spatial (calculated from Table 1) and temporal (calculated from Figure 5) variability of CH4
fluxes and CO2 fluxes of aquatic sites (a) and dry sites (b) accordingly. Temporal variability of CH4 fluxes from terrestrial sites was
285
not calculated. 3.4 Comparison of spatial and temporal variability
275 We calculated coefficients of variation (CV) to make spatial and temporal variability comparable (Figure 6). Consistent with
our hypothesis, spatial variability of CH4 emissions had a higher CV compared to CO2 both in aquatic and dry habitats. Also
consistent with our hypothesis, temporal variability of the CO2 flux had a higher CV than CH4 flux at the aquatic sites. At the
dry sites (where we did not measure temporal changes in CH4 flux) the temporal variability of the CO2 flux also showed a high We calculated coefficients of variation (CV) to make spatial and temporal variability comparable (Figure 6). Consistent with
our hypothesis, spatial variability of CH4 emissions had a higher CV compared to CO2 both in aquatic and dry habitats. Also
consistent with our hypothesis, temporal variability of the CO2 flux had a higher CV than CH4 flux at the aquatic sites. At the
dry sites (where we did not measure temporal changes in CH4 flux) the temporal variability of the CO2 flux also showed a high CV. If we want to judge the consequences of this result on upscaling, however, the absolute height of the fluxes as well as the
280
relative areas of the different habitats need to be considered. CV. If we want to judge the consequences of this result on upscaling, however, the absolute height of the fluxes as well as the
280
relative areas of the different habitats need to be considered. 13 Figure 6: Coefficients of variation of spatial (calculated from Table 1) and temporal (calculated from Figure 5) variability of CH4
fluxes and CO2 fluxes of aquatic sites (a) and dry sites (b) accordingly. Temporal variability of CH4 fluxes from terrestrial sites was
not calculated. https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. Figure 6: Coefficients of variation of spatial (calculated from Table 1) and temporal (calculated from Figure 5) variability of CH4
fluxes and CO2 fluxes of aquatic sites (a) and dry sites (b) accordingly. Temporal variability of CH4 fluxes from terrestrial sites was
285
not calculated. Figure 6: Coefficients of variation of spatial (calculated from Table 1) and temporal (calculated from Figure 5) variability of CH
fluxes and CO2 fluxes of aquatic sites (a) and dry sites (b) accordingly. Temporal variability of CH4 fluxes from terrestrial sites w
85
not calculated. 3.5 Upscaling About 10 % of our study area was not covered by water (Table 1). Within the dry area, all three habitat types contributed
similarly to the total area. If we apply a “perfect” approach considering spatial variability of CH4 and both spatial and temporal About 10 % of our study area was not covered by water (Table 1). Within the dry area, all three habitat types contributed
similarly to the total area. If we apply a “perfect” approach considering spatial variability of CH4 and both spatial and temporal
variability for CO2, our study reach emitted 91 mol CH4 d-1 and 16962 mol CO2 d-1 (Table 3). Thus, CH4 contributed 5 % to
290
total CO2-eq emissions. GHG emissions were dominated by the aquatic habitats which contributed 91 % to the total emissions. About 10 % of our study area was not covered by water (Table 1). Within the dry area, all three habitat types contributed
similarly to the total area. If we apply a “perfect” approach considering spatial variability of CH4 and both spatial and temporal
variability for CO2, our study reach emitted 91 mol CH4 d-1 and 16962 mol CO2 d-1 (Table 3). Thus, CH4 contributed 5 % to
290
total CO2-eq emissions. GHG emissions were dominated by the aquatic habitats which contributed 91 % to the total emissions. CH4 emissions from terrestrial habits can be neglected, while at the aquatic habitats they contributed about 6 % to the CO2-eq
emissions. Table 3: Total GHG emissions from our studied reach quantified considering both spatial and temporal variability. For CO2
emissions we multiplied the median flux from the temporal data (Figure 5) by habitat areas (Table 1). For aquatic CO2 emissions,
295
the temporal median flux at the side was also applied to the other aquatic habitats. For CH4 emissions we used the spatially different
fluxes (Table 1). For CO2-eq the CH4 emissions were converted to CO2-eq using a GWP of 28 and added to the CO2 emissions. aquatic
terrestrial
all
middle
side
groyne
fields
total
sandy
muddy
vegetated
total
total
CH4 [mol d-1]
36.6
14.94
39
91
0.018
0.023
-0.016
0
91
CO2 [mol d-1]
9960
1992
3347
15299
425
137
1101
1663
16962
CO2-eq [mol d-1]
10333
2144
3747
16224
425
137
1101
1664
17887 variability for CO2, our study reach emitted 91 mol CH4 d-1 and 16962 mol CO2 d-1 (Table 3). 3.5 Upscaling Thus, CH4 contributed 5 % to
290
total CO2-eq emissions. GHG emissions were dominated by the aquatic habitats which contributed 91 % to the total emissions. CH4 emissions from terrestrial habits can be neglected, while at the aquatic habitats they contributed about 6 % to the CO2-eq
emissions. Table 3: Total GHG emissions from our studied reach quantified considering both spatial and temporal variability. For CO2
emissions we multiplied the median flux from the temporal data (Figure 5) by habitat areas (Table 1). For aquatic CO2 emissions,
295
the temporal median flux at the side was also applied to the other aquatic habitats. For CH4 emissions we used the spatially different
fluxes (Table 1). For CO2-eq the CH4 emissions were converted to CO2-eq using a GWP of 28 and added to the CO2 emissions. aquatic
terrestrial
all
middle
side
groyne
fields
total
sandy
muddy
vegetated
total
total
CH4 [mol d-1]
36.6
14.94
39
91
0.018
0.023
-0.016
0
91
CO2 [mol d-1]
9960
1992
3347
15299
425
137
1101
1663
16962 aquatic
terrestrial
all
middle
side
groyne
fields
total
sandy
muddy
vegetated
total
total
CH4 [mol d-1]
36.6
14.94
39
91
0.018
0.023
-0.016
0
91
CO2 [mol d-1]
9960
1992
3347
15299
425
137
1101
1663
16962
CO2-eq [mol d-1]
10333
2144
3747
16224
425
137
1101
1664
17887 14 https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. 4.1 Spatial variability
300 Concentrations were lower in the
groyne fields, most probably because of higher photosynthetic activity and higher plankton biomass (Pusch and Fischer, 2006). However, different CO2 concentrations did not translate into spatial differences in CO2 emissions, because higher CO2
315
concentrations were accompanied by lower gas transfer velocities. This highlights the interacting role of both concentration
and gas transfer velocity in shaping spatial patterns of GHG emissions from rivers. While spatial differences in concentrations have already been acknowledged (Bussmann et al., 2022) our study is, to our
knowledge, the first to show small-scale spatial differences in gas transfer velocities in a river. Measuring k600 in rivers is not knowledge, the first to show small-scale spatial differences in gas transfer velocities in a river. Measuring k600 in rivers is not
an easy task, because tracer addition approaches, as typically used in small streams (Hope et al., 2001), cannot be applied. The
320
floating chamber method is probably the only existing method that can be used in intermediate to large streams (Lorke et al.,
2015). The spatial resolution of the method depends on the duration of the measurement, because the boat is drifting during
the measurement. We tried to minimise measuring time to about 2 minutes to optimise spatial resolution. Thus, at the measured
flow velocity of 0.8 m s-1, a typical flux measurement spanned a drift path of about 100m, which might be the limit for the an easy task, because tracer addition approaches, as typically used in small streams (Hope et al., 2001), cannot be applied. The
320
floating chamber method is probably the only existing method that can be used in intermediate to large streams (Lorke et al.,
2015). The spatial resolution of the method depends on the duration of the measurement, because the boat is drifting during
the measurement. We tried to minimise measuring time to about 2 minutes to optimise spatial resolution. Thus, at the measured
flow velocity of 0.8 m s-1, a typical flux measurement spanned a drift path of about 100m, which might be the limit for the spatial resolution in the direction of the flow. Since we were drifting parallel to the shore, however, the method was well suited
325
to distinguish k600 between the middle and side of the river. While higher k600 values at the side of the river were expected, the
high k600 in the groyne fields are somehow surprising. 4.1 Spatial variability
300 Our results show considerable spatial variability of CH4 emissions from aquatic sites. This confirms earlier observations
(Staniek, 2018) and can be explained by the fact that CH4 is primarily produced in the sediment and thus, depends on the
spatial heterogeneity of the sediments. Sediment deposition in rivers is highly heterogeneous and depends on hydrodynamics
(Henning and Hentschel, 2013). Fine material rich in organic matter preferably settles at low stream velocity, in our case in the groyne fields. Thus, our study confirms the hypothesis that the groyne fields are the major source of CH4 in river Elbe
305
(Bussmann et al., 2022). The data also revealed a significant difference between aquatic and terrestrial CH4 emissions. CH4 emissions from dry
sediments have rarely been measured, and there is still debate about their significance. A recent global survey indicates that
these CH4 emissions probably cannot be neglected (Paranaíba et al., 2021). However, our results confirm previous observations the groyne fields. Thus, our study confirms the hypothesis that the groyne fields are the major source of CH4 in river Elbe
305
(Bussmann et al., 2022). The data also revealed a significant difference between aquatic and terrestrial CH4 emissions. CH4 emissions from dry
sediments have rarely been measured, and there is still debate about their significance. A recent global survey indicates that
these CH4 emissions probably cannot be neglected (Paranaíba et al., 2021). However, our results confirm previous observations that CH4 emissions from dry river sediments are rather small compared to CO2 emissions (Koschorreck et al., 2022). It has
310
been suggested that there might be local hot spots of CH4 emissions (Marcé et al., 2019), but our measurements did not show
any evidence for such hot spots. We also observed considerable spatial variability in the CO2 concentration in the water. Concentrations were lower in the
groyne fields, most probably because of higher photosynthetic activity and higher plankton biomass (Pusch and Fischer, 2006). that CH4 emissions from dry river sediments are rather small compared to CO2 emissions (Koschorreck et al., 2022). It has
310
been suggested that there might be local hot spots of CH4 emissions (Marcé et al., 2019), but our measurements did not show
any evidence for such hot spots. We also observed considerable spatial variability in the CO2 concentration in the water. 4.1 Spatial variability
300 However, we argue that this bias might be small in our study when CO2 uptake by the
plants during the day was probably largely compensated by higher CO2 emissions due to plant respiration during the night. Taken together, our results show that spatial differences were especially apparent for CH4. The terrestrial habitats need to be
350
considered for CO2 emissions while they can probably be neglected for ecosystem-scale CH4 emissions from river Elbe. Taken together, our results show that spatial differences were especially apparent for CH4. The terrestrial habitats need to be
350
considered for CO2 emissions while they can probably be neglected for ecosystem-scale CH4 emissions from river Elbe. 4.1 Spatial variability
300 However, the observed trend of decreasing CO2 emissions with increasing sediment moisture (Figure 5) shows that
wetter conditions not necessarily result in higher fluxes. CO2 emissions from muddy sediments obviously result from a complex interplay between organic matter availability and moisture dependent gas transport limitation (Keller et al., 2020). 340
The sediment in the proximity of terrestrial vegetation showed clearly elevated CO2 emissions, confirming earlier observations
(Bolpagni et al., 2017; Koschorreck et al., 2022). Since we excluded plants from our chambers, these elevated CO2 emissions
are probably caused by root respiration. It is well known that in soils root respiration contributes about 50 % to soil respiration
(Hanson et al., 2000). It is clear that at vegetated sites our CO2 fluxes cannot be equated with net ecosystem exchange because complex interplay between organic matter availability and moisture dependent gas transport limitation (Keller et al., 2020). 340
The sediment in the proximity of terrestrial vegetation showed clearly elevated CO2 emissions, confirming earlier observations
(Bolpagni et al., 2017; Koschorreck et al., 2022). Since we excluded plants from our chambers, these elevated CO2 emissions
are probably caused by root respiration. It is well known that in soils root respiration contributes about 50 % to soil respiration
(Hanson et al., 2000). It is clear that at vegetated sites our CO2 fluxes cannot be equated with net ecosystem exchange because the plants were excluded from our chambers. To fully assess the effect of terrestrial plants on river CO2 emissions,
345
measurements using transparent chambers and considering light conditions as well as plant biomass determinations are
necessary. The exclusion of plants from our measurements means that we systematically overestimate CO2 emissions from
vegetated sites in the growing season. However, we argue that this bias might be small in our study when CO2 uptake by the
plants during the day was probably largely compensated by higher CO2 emissions due to plant respiration during the night. the plants were excluded from our chambers. To fully assess the effect of terrestrial plants on river CO2 emissions,
345
measurements using transparent chambers and considering light conditions as well as plant biomass determinations are
necessary. The exclusion of plants from our measurements means that we systematically overestimate CO2 emissions from
vegetated sites in the growing season. 4.1 Spatial variability
300 This indicates that factors like bottom roughness and flow energy
dissipation at the banks had a larger effect on turbulence on k than simply flow velocity (which was higher in the middle of
the stream). Wind was not an important factor controlling k600 values in our study because wind speeds were low and rather constant. It can be expected that in larger and winding rivers with variable fetch, wind field heterogeneities further contribute
330
to the spatial variability of k600. It is reasonable to assume that spatially variable gas transfer velocities should also be 15 https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. considered in stream metabolism calculations where k600 is used to quantify oxygen exchange between water and atmosphere
(Demars et al., 2015). Compared to the aquatic sites, the CO2 emissions from terrestrial sites showed considerable inter-habitat variability. Higher considered in stream metabolism calculations where k600 is used to quantify oxygen exchange between water and atmosphere
(Demars et al., 2015). C
d
h
i
i
h CO
i i
f
i l i
h
d
id
bl i
h bi
i bili
Hi h Compared to the aquatic sites, the CO2 emissions from terrestrial sites showed considerable inter-habitat variability. Higher
CO2 emissions from darker (= more muddy) sites compared to sandy sites have been used to scale up CO2 emissions from dry
335
river sediments using remote sensing (Mallast et al., 2020). They can be explained both by higher organic matter content of
the muddy sediments as well as higher sediment moisture which favors microbial CO2 production in the sediment (Keller et
al., 2020). However, the observed trend of decreasing CO2 emissions with increasing sediment moisture (Figure 5) shows that
wetter conditions not necessarily result in higher fluxes. CO2 emissions from muddy sediments obviously result from a
l
i
l
b
i
il bili
d
i
d
d
li i
i
(K ll
l 2020)
340 335 CO2 emissions from darker (= more muddy) sites compared to sandy sites have been used to scale up CO2 emissions from dry
335
river sediments using remote sensing (Mallast et al., 2020). They can be explained both by higher organic matter content of
the muddy sediments as well as higher sediment moisture which favors microbial CO2 production in the sediment (Keller et
al., 2020). 4.2 Temporal variability of CO2 and CH4 Our results confirm that diurnal variability of CO2, which has been shown in streams (Attermeyer et al., 2021; Gómez-Gener
et al., 2021) as well as marine systems (Honkanen et al., 2021), is also relevant in rivers. Interestingly, the shape of the diurnal
curve of CO2 emissions differed between habitats, showing that different regulatory mechanisms are at play. 355 Our results confirm that diurnal variability of CO2, which has been shown in streams (Attermeyer et al., 2021; Gómez-Gener
et al., 2021) as well as marine systems (Honkanen et al., 2021), is also relevant in rivers. Interestingly, the shape of the diurnal
curve of CO2 emissions differed between habitats, showing that different regulatory mechanisms are at play. 355
At aquatic sites biological fixation and mineralization of carbon led to sinusoidal diurnal pCO2 variations, with a maximum in
the morning and a minimum in the afternoon. This pattern was obviously driven by light since the diurnal temperature
amplitude in the water was below 1.5°C. The temporal pattern at the muddy site was also most likely driven by the interplay between microalgae primary production Our results confirm that diurnal variability of CO2, which has been shown in streams (Atterm
et al., 2021) as well as marine systems (Honkanen et al., 2021), is also relevant in rivers. Intere Our results confirm that diurnal variability of CO2, which has been shown in streams (Attermeyer et al., 2021; Gómez-Gener
et al., 2021) as well as marine systems (Honkanen et al., 2021), is also relevant in rivers. Interestingly, the shape of the diurnal curve of CO2 emissions differed between habitats, showing that different regulatory mechanisms are at play. 355
At aquatic sites biological fixation and mineralization of carbon led to sinusoidal diurnal pCO2 variations, with a maximum in
the morning and a minimum in the afternoon. This pattern was obviously driven by light since the diurnal temperature
amplitude in the water was below 1.5°C. The temporal pattern at the muddy site was also most likely driven by the interplay between microalgae primary production curve of CO2 emissions differed between habitats, showing that different regulatory mechanisms are at play. 355
At aquatic sites biological fixation and mineralization of carbon led to sinusoidal diurnal pCO2 variations, with a maximum in
the morning and a minimum in the afternoon. This pattern was obviously driven by light since the diurnal temperature
amplitude in the water was below 1.5°C. 4.2 Temporal variability of CO2 and CH4 Methane is produced in deeper sediment layers and, thus, is not affected by light
driven changes of redox conditions at the very sediment surface. Even if CH4 oxidation at the sediment surface were affected by phototrophic activity, this would not result in fluctuating emissions because these are buffered by the CH4 pool in the water. 385
Thus, we have confirmed our hypothesis that diurnal variability is relevant for CO2 but not for CH4. We also show that the
shape of the diurnal CO2 flux curve depends on the habitat. As already acknowledged in the literature, this has important
implications for monitoring strategies and upscaling (Gómez-Gener et al., 2021). by phototrophic activity, this would not result in fluctuating emissions because these are buffered by the CH4 pool in the water. 385
Thus, we have confirmed our hypothesis that diurnal variability is relevant for CO2 but not for CH4. We also show that the
shape of the diurnal CO2 flux curve depends on the habitat. As already acknowledged in the literature, this has important
implications for monitoring strategies and upscaling (Gómez-Gener et al., 2021). 4.2 Temporal variability of CO2 and CH4 The reason for a “Plateau behavior” was the migration of
some cells to greater depth in order to avoid too much light. Possible physiological explanations might be the existence of
alternative electron sinks (e g the Mehler reaction or photorespiration) or limitation by Calvin cycle reactions
375 similar fast switching process between plateau-like CO2 production in the dark and CO2 uptake during the light, was observed
370
(Tang and Kristensen, 2007). Detailed investigations of benthic primary production on exposed marine sediments showed that
both linear and plateau relationships were obtained between the fluorescence parameter (relative electron transport rate [rETR])
and the community-level carbon-fixation rate (Migne et al., 2007). The reason for a “Plateau behavior” was the migration of
some cells to greater depth in order to avoid too much light. Possible physiological explanations might be the existence of alternative electron sinks (e.g., the Mehler reaction or photorespiration) or limitation by Calvin cycle reactions. 375
At vegetated sites, the diurnal pattern was probably driven by diurnal fluctuating plant metabolism and root respiration. However, the diurnal CO2 flux curve was not in phase with the light or temperature curve. This can be explained by a hysteresis
effect caused by the transit time of CO2 from the source of its formation (probably the plant roots) and the sediment surface
(Koschorreck et al., 2022; Phillips et al., 2011). Diurnal variability of CH4 emissions was observed in a study focussing on spatio-temporal variability of GHG emissions in
380
the Danube Delta (Canning et al., 2021). Elevated CH4 emissions from a floodplain lake and a channel were attributed to
stratification and temporary mixing of the water column. In our case the water column was permanently mixed and diurnal
variability was not an issue for CH4 fluxes. Methane is produced in deeper sediment layers and, thus, is not affected by light
driven changes of redox conditions at the very sediment surface. Even if CH4 oxidation at the sediment surface were affected Diurnal variability of CH4 emissions was observed in a study focussing on spatio-temporal variability of GHG emissions in
380
the Danube Delta (Canning et al., 2021). Elevated CH4 emissions from a floodplain lake and a channel were attributed to
stratification and temporary mixing of the water column. In our case the water column was permanently mixed and diurnal
variability was not an issue for CH4 fluxes. 4.2 Temporal variability of CO2 and CH4 The temporal pattern at the muddy site was also most likely driven by the interplay between microalgae primary production The temporal pattern at the muddy site was also most likely driven by the interplay between microalgae primary production
and respiration, but the shape of the diurnal CO2 curve differed considerably. These data nicely demonstrate that the same
360
regulatory mechanism (light-dependent balance of photosynthesis and respiration) may result in different diurnal pattern
depending on the physical environments. In water, changes in biological activity are buffered by the DIC pool in the water,
resulting in gradual changes of CO2 concentration during the day. At terrestrial sites, switching photosynthesis on and off and respiration, but the shape of the diurnal CO2 curve differed considerably. These data nicely demonstrate that the same
360
regulatory mechanism (light-dependent balance of photosynthesis and respiration) may result in different diurnal pattern
depending on the physical environments. In water, changes in biological activity are buffered by the DIC pool in the water,
resulting in gradual changes of CO2 concentration during the day. At terrestrial sites, switching photosynthesis on and off 16 https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. changes the sediment from a CO2 sink to a source and back again. Since microbial respiration depends on temperature and
temperature fluctuations in the sediment were quite large, it is somewhat surprising that we did not see a pronounced
365
temperature signal in the CO2 flux (as in Koschorreck et al. (2022)). A possible explanation is that the CO2 pool in the pore
space buffers the effect of fluctuating respiration. Since the flux of CO2 between the sediment and the atmosphere is driven by
the concentration gradient, this results in rather constant CO2 efflux during the night. During the day this efflux is most
probably blocked by photosynthetic uptake of CO2 by benthic microalgae. In a laboratory study with marine sediments, a similar fast switching process between plateau-like CO2 production in the dark and CO2 uptake during the light, was observed
370
(Tang and Kristensen, 2007). Detailed investigations of benthic primary production on exposed marine sediments showed that
both linear and plateau relationships were obtained between the fluorescence parameter (relative electron transport rate [rETR])
and the community-level carbon-fixation rate (Migne et al., 2007). 4.3 Implications for measurement strategy and upscaling Thus, if stream width is not explicitly measured, expected errors can become considerably larger than the mean width of our reach of 200 m, which we obtain by dividing the water surface area (Table 1) by reach length (0.96 km). Although we found good agreement between measured widths and those calculated using the equation of Raymond et al. (2012), measured flux widths should always be used for field studies because of the large scatter in the regression and the
logyrhytmic scale used. Thus, if stream width is not explicitly measured, expected errors can become considerably larger than
the standard error. These approaches, however, do not include dry sediment areas. In our case 9 % of the total area was dry,
400
which is somewhat lower than the 26 % estimated from remote sensing data during the extreme drought in 2018 (Mallast et
al., 2020). During that drought, dry sediment areas showed clear longitudinal variability along the river depending on
topography ranging from 2 % to 40 % of the river area being dry. A straightforward strategy would be to determine dry areas
for different discharge scenarios to derive a quantitative relation between dry area, water area, and discharge. the standard error. These approaches, however, do not include dry sediment areas. In our case 9 % of the total area was dry,
400
which is somewhat lower than the 26 % estimated from remote sensing data during the extreme drought in 2018 (Mallast et
al., 2020). During that drought, dry sediment areas showed clear longitudinal variability along the river depending on
topography ranging from 2 % to 40 % of the river area being dry. A straightforward strategy would be to determine dry areas
for different discharge scenarios to derive a quantitative relation between dry area, water area, and discharge. the standard error. These approaches, however, do not include dry sediment areas. In our case 9 % of the total area was dry,
400
which is somewhat lower than the 26 % estimated from remote sensing data during the extreme drought in 2018 (Mallast et
al., 2020). During that drought, dry sediment areas showed clear longitudinal variability along the river depending on
topography ranging from 2 % to 40 % of the river area being dry. A straightforward strategy would be to determine dry areas
for different discharge scenarios to derive a quantitative relation between dry area, water area, and discharge. 4.3 Implications for measurement strategy and upscaling Based on our results, the best monitoring strategy for our stream reach should consider spatial variability of CH4 and both
390
spatial and temporal variability of CO2. Applying that approach to our data (Table 3) revealed the dominant role of CO2 (due
to low CH4 fluxes) and aquatic habitats (due to their larger area). It is evident that the exact quantification of habitat areas is crucial. Stream surface areas are typically estimated from empirical
relations depending on stream order (Raymond et al., 2012) or remote sensing (Palmer and Ruhi, 2018). Estimating river width Based on our results, the best monitoring strategy for our stream reach should consider spatial variability of CH4 and both
390
spatial and temporal variability of CO2. Applying that approach to our data (Table 3) revealed the dominant role of CO2 (due
to low CH4 fluxes) and aquatic habitats (due to their larger area). It is evident that the exact quantification of habitat areas is crucial. Stream surface areas are typically estimated from empirical It is evident that the exact quantification of habitat areas is crucial. Stream surface areas are typically estimated from empirical
relations depending on stream order (Raymond et al., 2012) or remote sensing (Palmer and Ruhi, 2018). Estimating river width
from annual mean discharge, according to Raymond et al. (2012), reveals in our case a width of 183m. This is very similar to
395 relations depending on stream order (Raymond et al., 2012) or remote sensing (Palmer and Ruhi, 2018). Estimating river width
from annual mean discharge, according to Raymond et al. (2012), reveals in our case a width of 183m. This is very similar to
395 from annual mean discharge, according to Raymond et al. (2012), reveals in our case a width of 183m. This is very similar to
395 17 https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. the mean width of our reach of 200 m, which we obtain by dividing the water surface area (Table 1) by reach length (0.96 km). Although we found good agreement between measured widths and those calculated using the equation of Raymond et al. (2012), measured flux widths should always be used for field studies because of the large scatter in the regression and the
logyrhytmic scale used. 4.3 Implications for measurement strategy and upscaling For the design of a perfect
monitoring strategy for a given river, the particular habitat types and diversity need to be considered. We can also expect that
the role of spatial and temporal variability changes with the season, both because habitat areas and regulatory factors like
420
temperature or day-length change (Koschorreck et al., 2022). We would also expect that CH4 variability needs to be re-assessed
if ebullition becomes relevant (Maeck et al., 2014), especially in dammed river sections (Matoušů et al., 2019) or floodplain
waters and under warm conditions (Barbosa et al., 2021). More natural river-floodplain systems containing floodplain lakes
are known to harbour extreme spatial variability with significant CH4 emissions (Maier et al., 2021), calling for a more
sophisticated monitoring approach (Canning et al., 2021). 425 g
gy
g
p
yp
y
p
the role of spatial and temporal variability changes with the season, both because habitat areas and regulatory factors like
420
temperature or day-length change (Koschorreck et al., 2022). We would also expect that CH4 variability needs to be re-assessed
if ebullition becomes relevant (Maeck et al., 2014), especially in dammed river sections (Matoušů et al., 2019) or floodplain
waters and under warm conditions (Barbosa et al., 2021). More natural river-floodplain systems containing floodplain lakes
are known to harbour extreme spatial variability with significant CH4 emissions (Maier et al., 2021), calling for a more
sophisticated monitoring approach (Canning et al., 2021). 425 sophisticated monitoring approach (Canning et al., 2021). 425 Competing Interests The contact author has declared that none of the authors has any competing interests The contact author has declared that none of the authors has any competing interests 4.4 Conclusion Although we only provide a snapshot case study at a German river, we can derive a number of conclusions relevant for the
quantification of GHG emission from large temperate rivers. It is now evident from several studies that continuous measurements are necessary to come up with realistic emission It is now evident from several studies that continuous measurements are necessary to come up with realistic emission
approaches. CO2 probes are becoming more and more popular. Deploying them in numerous rivers will improve global riverine
430
CO2 emissions estimates. It is now evident from several studies that continuous measurements are necessary to come up with realistic emission
approaches. CO2 probes are becoming more and more popular. Deploying them in numerous rivers will improve global riverine
430
CO2 emissions estimates. It is also clear that river sediments drying up at low discharge need to be considered at least for CO2 budgets. However, when approaches. CO2 probes are becoming more and more popular. Deploying them in numerous rivers will improve global riverine
430
CO2 emissions estimates. It is also clear that river sediments drying up at low discharge need to be considered at least for CO2 budgets. However, when
it comes to total GHG emissions, lower CH4 emissions compensate for higher CO2 emissions from dry sediments; this is a
scenario already hypothesised for reservoir sediments (Marcé et al., 2019). CO2 emissions estimates. It is also clear that river sediments drying up at low discharge need to be considered at least for CO2 budgets. However, when
it comes to total GHG emissions, lower CH4 emissions compensate for higher CO2 emissions from dry sediments; this is a
scenario already hypothesised for reservoir sediments (Marcé et al., 2019). Finally, our data show that anthropogenic modification of the river (here: the construction of groynes) has the potential to alter
435
GHG emissions significantly. In our case, the groyne fields nearly doubled CH4 emissions from the river. 4.3 Implications for measurement strategy and upscaling We used our dataset to simulate different monitoring approaches and compared them with the optimal approach (Figure 7). 405
Only sampling the river during the day at the side would result in more than 50 % under estimation of the real GHG emissions
– mostly because higher CO2 emissions during the night are not considered. Measuring in the middle of the river or even in all
habitats during the day would only slightly improve the result. If both CO2 and CH4 emissions were measured over a 24 h
cycle only along the side of the river, we would slightly overestimate emissions because of high CH4 emissions along the side. The convenient approach of deploying only a CO2 probe at the side of the river would result in about 5 % underestimation of
410
total CO2-eq emissions from our study reach. Figure 7: Deviation of total GHG emissions (CO2-eq) obtained by different monitoring approaches from perfect approach (spatial
variability of CH4 and both spatial and temporal variability of CO2 considered). 415 Figure 7: Deviation of total GHG emissions (CO2-eq) obtained by different monitoring approaches from perfect approach (spatial
variability of CH4 and both spatial and temporal variability of CO2 considered). 415 Figure 7: Deviation of total GHG emissions (CO2-eq) obtained by different monitoring approaches from perfect approach (spatial
variability of CH4 and both spatial and temporal variability of CO2 considered). 415 18 https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. However, these considerations differ depending on the target gas. If we sample only during the day along the side of the river,
for example, we would underestimate CO2 emissions by about 50 % but overestimate CH4 emissions by 40 % (Figure S8). Although our results reveal some general principles, they cannot be simply applied to other systems. For the design of a perfect
monitoring strategy for a given river, the particular habitat types and diversity need to be considered. We can also expect that However, these considerations differ depending on the target gas. If we sample only during the day along the side of the river,
for example, we would underestimate CO2 emissions by about 50 % but overestimate CH4 emissions by 40 % (Figure S8). Although our results reveal some general principles, they cannot be simply applied to other systems. References Bolpagni, R., Folegot, S., Laini, A., and Bartoli, M.: Role of ephemeral vegetation of emerging river bottoms in modulating CO2 exchanges
across a temperate large lowland river stretch, Aquat Sci, 79, 149-158, 10.1007/s00027-016-0486-z, 2017. p
g
q
Bolpagni, R., Laini, A., Mutti, T., Viaroli, P., and Bartoli, M.: Connectivity and habitat typology drive CO2 and CH4 fluxes across land–
water interfaces in lowland rivers, Ecohydrology, 12, e2036, doi:10.1002/eco.2036, 2019. 465 Bolpagni, R., Laini, A., Mutti, T., Viaroli, P., and Bartoli, M.: Connectivity and habitat typology drive CO2 and CH4 fluxes across land–
water interfaces in lowland rivers, Ecohydrology, 12, e2036, doi:10.1002/eco.2036, 2019. 465
Bussmann, I., Koedel, U., Schütze, C., Kamjunke, N., and Koschorreck, M.: Spatial Variability and Hotspots of Methane Concentrations in Bolpagni, R., Laini, A., Mutti, T., Viaroli, P., and Bartoli, M.: Connectivity and habitat typology drive CO2 and CH4 fluxes across land
water interfaces in lowland rivers, Ecohydrology, 12, e2036, doi:10.1002/eco.2036, 2019. 465
Bussmann, I., Koedel, U., Schütze, C., Kamjunke, N., and Koschorreck, M.: Spatial Variability and Hotspots of Methane Concentrations in
a Large Temperate River, Frontiers in Environmental Science, 10, 10.3389/fenvs.2022.833936, 2022. Canning, A., Wehrli, B., and Körtzinger, A.: Methane in the Danube Delta: the importance of spatial patterns and diel cycles for atmospheric
emission estimates, Biogeosciences, 18, 3961-3979, 10.5194/bg-18-3961-2021, 2021. y
gy
tze, C., Kamjunke, N., and Koschorreck, M.: Spatial Variability and Hotspots of Methane Concentrations in
tiers in Environmental Science, 10, 10.3389/fenvs.2022.833936, 2022. a Large Temperate River, Frontiers in Environmental Science, 10, 10.3389/fenvs.2022.833936, 2022. Canning, A., Wehrli, B., and Körtzinger, A.: Methane in the Danube Delta: the importance of spatial patterns and diel cycles for atmospheric
emission estimates, Biogeosciences, 18, 3961-3979, 10.5194/bg-18-3961-2021, 2021. 4 0 ,
g
,
,
,
g
,
Demars, B. O. L., Thompson, J., and Manson, J. R.: Stream metabolism and the open diel oxygen method: Principles, practice, and
470
perspectives, Limnol Oceanogr-Meth, 13, 356-374, 10.1002/lom3.10030, 2015. g
son, J., and Manson, J. R.: Stream metabolism and the open diel oxygen method: Principles, practice, an
ogr-Meth, 13, 356-374, 10.1002/lom3.10030, 2015. Demars, B. O. L., Thompson, J., and Manson, J. R.: Stream metabolism and the open diel oxygen method: Principles, practice, and
470
perspectives, Limnol Oceanogr-Meth, 13, 356-374, 10.1002/lom3.10030, 2015. Gómez-Gener, L., Obrador, B., von Schiller, D., Marce, R., Casas-Ruiz, J. References Attermeyer, K., Casas-Ruiz, J. P., Fuss, T., Pastor, A., Cauvy-Fraunié, S., Sheath, D., Nydahl, A. C., Doretto, A., Portela, A. P., Doyle, B. C., Simov, N., Gutmann Roberts, C., Niedrist, G. H., Timoner, X., Evtimova, V., Barral-Fraga, L., Bašić, T., Audet, J., Deininger, A., Busst,
G., Fenoglio, S., Catalán, N., de Eyto, E., Pilotto, F., Mor, J.-R., Monteiro, J., Fletcher, D., Noss, C., Colls, M., Nagler, M., Liu, L., Romero
450
González-Quijano, C., Romero, F., Pansch, N., Ledesma, J. L. J., Pegg, J., Klaus, M., Freixa, A., Herrero Ortega, S., Mendoza-Lera, C.,
Bednařík, A., Fonvielle, J. A., Gilbert, P. J., Kenderov, L. A., Rulík, M., and Bodmer, P.: Carbon dioxide fluxes increase from day to night
across European streams, Communications Earth & Environment, 2, 118, 10.1038/s43247-021-00192-w, 2021. Attermeyer, K., Casas-Ruiz, J. P., Fuss, T., Pastor, A., Cauvy-Fraunié, S., Sheath, D., Nydahl, A. C., Doretto, A., Portela, A. P., Doyle, B. C., Simov, N., Gutmann Roberts, C., Niedrist, G. H., Timoner, X., Evtimova, V., Barral-Fraga, L., Bašić, T., Audet, J., Deininger, A., Busst,
G., Fenoglio, S., Catalán, N., de Eyto, E., Pilotto, F., Mor, J.-R., Monteiro, J., Fletcher, D., Noss, C., Colls, M., Nagler, M., Liu, L., Romero
450 Attermeyer, K., Casas-Ruiz, J. P., Fuss, T., Pastor, A., Cauvy-Fraunié, S., Sheath, D., Nydahl, A. C., Doretto, A., Portela, A. P., Doyle, B. C., Simov, N., Gutmann Roberts, C., Niedrist, G. H., Timoner, X., Evtimova, V., Barral-Fraga, L., Bašić, T., Audet, J., Deininger, A., Busst, G., Fenoglio, S., Catalán, N., de Eyto, E., Pilotto, F., Mor, J.-R., Monteiro, J., Fletcher, D., Noss, C., Colls, M., Nagler, M., Liu, L., Romero
450
González-Quijano, C., Romero, F., Pansch, N., Ledesma, J. L. J., Pegg, J., Klaus, M., Freixa, A., Herrero Ortega, S., Mendoza-Lera, C.,
Bednařík, A., Fonvielle, J. A., Gilbert, P. J., Kenderov, L. A., Rulík, M., and Bodmer, P.: Carbon dioxide fluxes increase from day to night
across European streams, Communications Earth & Environment, 2, 118, 10.1038/s43247-021-00192-w, 2021. Barbosa, P. M., Melack, J. M., Amaral, J. H. F., Linkhorst, A., and Forsberg, B. R.: Large Seasonal and Habitat Differences in Methane G., Fenoglio, S., Catalán, N., de Eyto, E., Pilotto, F., Mor, J.-R., Monteiro, J., Fletcher, D., Noss, C., Colls, M., Nagler, M., Liu, L., Romero
450
González-Quijano, C., Romero, F., Pansch, N., Ledesma, J. L. J., Pegg, J., Klaus, M., Freixa, A., Herrero Ortega, S., Mendoza-Lera, C.,
Bednařík, A., Fonvielle, J. References P., Proia, L., Acuna, V., Catalan, N., Munoz, I., and Koschorreck,
M.: Hot spots for carbon emissions from Mediterranean fluvial networks during summer drought, Biogeochemistry, 125, 409-426,
10.1007/s10533-015-0139-7, 2015. p
p
,
g
,
,
,
,
Gómez-Gener, L., Obrador, B., von Schiller, D., Marce, R., Casas-Ruiz, J. P., Proia, L., Acuna, V., Catalan, N., Munoz, I., and Koschorreck,
M.: Hot spots for carbon emissions from Mediterranean fluvial networks during summer drought, Biogeochemistry, 125, 409-426,
10.1007/s10533-015-0139-7, 2015. Gómez-Gener, L., Rocher-Ros, G., Battin, T., Cohen, M. J., Dalmagro, H. J., Dinsmore, K. J., Drake, T. W., Duvert, C., Enrich-Prast, A.,
475
Horgby, Å., Johnson, M. S., Kirk, L., Machado-Silva, F., Marzolf, N. S., McDowell, M. J., McDowell, W. H., Miettinen, H., Ojala, A. K., ener, L., Rocher-Ros, G., Battin, T., Cohen, M. J., Dalmagro, H. J., Dinsmore, K. J., Drake, T. W., Duvert,
Å., Johnson, M. S., Kirk, L., Machado-Silva, F., Marzolf, N. S., McDowell, M. J., McDowell, W. H., Miettin Gómez-Gener, L., Rocher-Ros, G., Battin, T., Cohen, M. J., Dalmagro, H. J., Dinsmore, K. J., Drake, T. W., Duvert, C., Enrich-Prast, A.,
475
Horgby, Å., Johnson, M. S., Kirk, L., Machado-Silva, F., Marzolf, N. S., McDowell, M. J., McDowell, W. H., Miettinen, H., Ojala, A. K.,
Peter, H., Pumpanen, J., Ran, L., Riveros-Iregui, D. A., Santos, I. R., Six, J., Stanley, E. H., Wallin, M. B., White, S. A., and Sponseller, R. A.: Global carbon dioxide efflux from rivers enhanced by high nocturnal emissions, Nature Geoscience, 14, 289-294, 10.1038/s41561-021-
00722-3, 2021. Peter, H., Pumpanen, J., Ran, L., Riveros-Iregui, D. A., Santos, I. R., Six, J., Stanley, E. H., Wallin, M. B., White, S. A., and Sponseller, R. A.: Global carbon dioxide efflux from rivers enhanced by high nocturnal emissions, Nature Geoscience, 14, 289-294, 10.1038/s41561-021-
00722-3, 2021. Hanson, P. J., Edwards, N. T., Garten, C. T., and Andrews, J. A.: Separating root and soil microbial contributions to soil respiration: A
480
review of methods and observations, Biogeochemistry, 48, 115-146, Doi 10.1023/A:1006244819642, 2000. i
d
h l
S di
i
d fl
i d
d b
l
d
difi d
h
i
lb
G Hanson, P. J., Edwards, N. T., Garten, C. T., and Andrews, J. A.: Separating root and soil microbial contributions to soil respiration: A
480
review of methods and observations, Biogeochemistry, 48, 115-146, Doi 10.1023/A:1006244819642, 2000. References A., Gilbert, P. J., Kenderov, L. A., Rulík, M., and Bodmer, P.: Carbon dioxide fluxes increase from day to night
across European streams, Communications Earth & Environment, 2, 118, 10.1038/s43247-021-00192-w, 2021. Barbosa, P. M., Melack, J. M., Amaral, J. H. F., Linkhorst, A., and Forsberg, B. R.: Large Seasonal and Habitat Differences in Methane
Ebullition
on
the
Amazon
Floodplain,
Journal
of
Geophysical
Research:
Biogeosciences,
126,
e2020JG005911,
455
https://doi.org/10.1029/2020JG005911, 2021. Barbosa, P. M., Melack, J. M., Amaral, J. H. F., Linkhorst, A., and Forsberg, B. R.: Large Seasonal and Habitat Differences in Methane
Ebullition
on
the
Amazon
Floodplain,
Journal
of
Geophysical
Research:
Biogeosciences,
126,
e2020JG005911,
455
https://doi.org/10.1029/2020JG005911, 2021. Bartoscheck, T., Fehrenbach, D., and Fehrenbach, J.: Das sensebook-Buch - 12 Projekte rund um Sensoren, dpunkt Verlag, Heidelberg,
2019. Ebullition
on
the
Amazon
Floodplain,
Journal
of
Geophysical
Research:
Biogeosciences,
126,
e2020JG005911,
455
https://doi.org/10.1029/2020JG005911, 2021. Bartoscheck, T., Fehrenbach, D., and Fehrenbach, J.: Das sensebook-Buch - 12 Projekte rund um Sensoren, dpunkt Verlag, Heidelberg,
2019. Battin, T. J., Lauerwald, R., Bernhardt, E. S., Bertuzzo, E., Gener, L. G., Hall, R. O., Hotchkiss, E. R., Maavara, T., Pavelsky, T. M., Ran, p
g
,
Bartoscheck, T., Fehrenbach, D., and Fehrenbach, J.: Das sensebook-Buch - 12 Projekte rund um Sensoren, dpunkt Verlag, Heidelberg,
2019. h, D., and Fehrenbach, J.: Das sensebook-Buch - 12 Projekte rund um Sensoren, dpunkt Verlag, Heidelber Battin, T. J., Lauerwald, R., Bernhardt, E. S., Bertuzzo, E., Gener, L. G., Hall, R. O., Hotchkiss, E. R., Maavara, T., Pavelsky, T. M., Ran,
L., Raymond, P., Rosentreter, J. A., and Regnier, P.: River ecosystem metabolism and carbon biogeochemistry in a changing world, Nature,
460
613, 449-459, 10.1038/s41586-022-05500-8, 2023. Bolpagni R Folegot S Laini A and Bartoli M : Role of ephemeral vegetation of emerging river bottoms in modulating CO2 exchanges L., Raymond, P., Rosentreter, J. A., and Regnier, P.: River ecosystem metabolism and carbon biogeochemistry in a changing world, Nature,
460
613, 449-459, 10.1038/s41586-022-05500-8, 2023. Bolpagni, R., Folegot, S., Laini, A., and Bartoli, M.: Role of ephemeral vegetation of emerging river bottoms in modulating CO2 exchanges
across a temperate large lowland river stretch, Aquat Sci, 79, 149-158, 10.1007/s00027-016-0486-z, 2017. Bolpagni, R., Laini, A., Mutti, T., Viaroli, P., and Bartoli, M.: Connectivity and habitat typology drive CO2 and CH4 fluxes across land– 613, 449-459, 10.1038/s41586-022-05500-8, 2023. Acknowledgements We would like to thank the captain of RV Albis, S. Bauth and our technician C. Völkner for help during fieldwork. Discharge
440
data were kindly provided by the German Federal Waterways and Shipping Administration (WSV) communicated by the
German Federal Institute of Hydrology (BfG). Thanks to Patrick Fink for providing light data and Peifang Leng for
commenting on the manuscript. We also thank P. Ronning for proofreading and his valuable comments and to Yao Li for
preparing the spatially distibuted ADCP data. This work was supported by funding from the Helmholtz Association in the 19 https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. framework of MOSES (Modular Observation Solutions for Earth Systems) and the POF-4 topic “Coastal Transition Zones
445
under Natural and Human Pressure”. framework of MOSES (Modular Observation Solutions for Earth Systems) and the POF-4 topic “Coastal Transition Zones
445
under Natural and Human Pressure”. References Henning, M., and Hentschel, B.: Sedimentation and flow patterns induced by regular and modified groynes on the River Elbe, Germany,
Ecohydrology, 6, 598-610, 10.1002/eco.1398, 2013. Honkanen M Müller J D Seppälä J Rehder G Kielosto S Ylöstalo P Mäkelä T Hatakka J and Laakso L : The diurnal cycle g
y
Henning, M., and Hentschel, B.: Sedimentation and flow patterns induced by regular and modified groynes on the River Elbe, Germany,
Ecohydrology, 6, 598-610, 10.1002/eco.1398, 2013. ,
,
,
,
pp
,
,
,
,
,
,
,
,
,
,
,
,
,
y
of pCO2 in the coastal region of the Baltic Sea, Ocean Sci., 17, 1657-1675, 10.5194/os-17-1657-2021, 2021. 485
Hope, D., Palmer, S. M., Billett, M. F., and Dawson, J. J. C.: Carbon dioxide and methane evasion from a temperate peatland stream, Limnol
Oceanogr, 46, 847-857, 2001. IPCC: Climate Change 2021: The Physical Science Basis. Contribution of Working Group I to the Sixth Assessment Report of the
Intergovernmental Panel on Climate Change, edited by: Masson-Delmotte, V., Zhai, P., Pirani, A., Connors, S. L., Péan, C., Berger, S., p
g
,
,
,
,
,
Hope, D., Palmer, S. M., Billett, M. F., and Dawson, J. J. C.: Carbon dioxide and methane evasion from a temperate peatland stream, Limnol
Oceanogr, 46, 847-857, 2001. IPCC: Climate Change 2021: The Physical Science Basis. Contribution of Working Group I to the Sixth Assessment Report of the p
g
Hope, D., Palmer, S. M., Billett, M. F., and Dawson, J. J. C.: Carbon dioxide and methane evasion from a temperate peatland stream, Limnol
Oceanogr, 46, 847-857, 2001. Caud, N., Chen, Y., Goldfarb, L., Gomis, M. I., Huang, M., Leitzell, K., Lonnoy, E., Matthews, J. B. R., Maycock, T. K., Waterfield, T.,
490
Yelekçi, O., Yu, R., and Zhou, B., Cambridge University Press, Cambridge, United Kingdom and New York, NY, USA, 2021. Keller, P. S., Catalán, N., von Schiller, D., Grossart, H. P., Koschorreck, M., Obrador, B., Frassl, M. A., Karakaya, N., Barros, N., Howitt,
J. A., Mendoza-Lera, C., Pastor, A., Flaim, G., Aben, R., Riis, T., Arce, M. I., Onandia, G., Paranaíba, J. R., Linkhorst, A., del Campo, R.,
Amado, A. M., Cauvy-Fraunié, S., Brothers, S., Condon, J., Mendonça, R. F., Reverey, F., Rõõm, E. I., Datry, T., Roland, F., Laas, A., Keller, P. S., Catalán, N., von Schiller, D., Grossart, H. P., Koschorreck, M., Obrador, B., Frassl, M. References p
g
j
Maeck, A., Hofmann, H., and Lorke, A.: Pumping methane out of aquatic sediments - ebullition forcing mechansims in an impounded river,
biogeosciences, 11, 2925-2938, 2014. g
Maier, M. S., Teodoru, C. R., and Wehrli, B.: Spatio-temporal variations in lateral and atmospheric carbon fluxes from the Danube Delta,
515
Biogeosciences, 18, 1417-1437, 10.5194/bg-18-1417-2021, 2021. Maier, M. S., Teodoru, C. R., and Wehrli, B.: Spatio-temporal variations in lateral and atmospheric carbon fluxes from the Danube Delta,
515
Biogeosciences, 18, 1417-1437, 10.5194/bg-18-1417-2021, 2021. Mallast, U., Staniek, M., and Koschorreck, M.: Spatial upscaling of CO2 emissions from exposed river sediments of the Elbe River during
an extreme drought, Ecohydrology, 13, ARTN e221610.1002/eco.2216, 2020. Marcé, R., Obrador, B., Gómez-Gener, L., Catalán, N., Koschorreck, M., Arce, M. I., Singer, G., and von Schiller, D.: Emissions from dry Biogeosciences, 18, 1417 1437, 10.5194/bg 18 1417 2021, 2021. Mallast, U., Staniek, M., and Koschorreck, M.: Spatial upscaling of CO2 emissions from exposed river sediments of the Elbe River during
an extreme drought, Ecohydrology, 13, ARTN e221610.1002/eco.2216, 2020. Marcé, R., Obrador, B., Gómez-Gener, L., Catalán, N., Koschorreck, M., Arce, M. I., Singer, G., and von Schiller, D.: Emissions from dry
inland
waters
are
a
blind
spot
in
the
global
carbon
cycle,
Earth-Science
Reviews,
188,
240-248,
520
https://doi.org/10.1016/j.earscirev.2018.11.012, 2019. Š oschorreck, M.: Spatial upscaling of CO2 emissions from exposed river sediments of the Elbe River during
ogy, 13, ARTN e221610.1002/eco.2216, 2020. g ,
y
gy,
,
,
Marcé, R., Obrador, B., Gómez-Gener, L., Catalán, N., Koschorreck, M., Arce, M. I., Singer, G., and von Schiller, D.: Emissions from dry
inland
waters
are
a
blind
spot
in
the
global
carbon
cycle,
Earth-Science
Reviews,
188,
240-248,
520
https://doi.org/10.1016/j.earscirev.2018.11.012, 2019. Matoušů, A., Rulík, M., Tušer, M., Bednařík, A., Šimek, K., and Bussmann, I.: Methane dynamics in a large river: a case study of the Elbe inland
waters
are
a
blind
spot
in
the
global
carbon
cycle,
Earth-Science
Reviews,
188,
240-248,
520
https://doi.org/10.1016/j.earscirev.2018.11.012, 2019. Matoušů, A., Rulík, M., Tušer, M., Bednařík, A., Šimek, K., and Bussmann, I.: Methane dynamics in a large river: a case study of the Elbe
River, Aquat Sci, 81, 12, 2019. Migne, A., Gevaert, F., Creach, A., Spilmont, N., Chevalier, E., and Davoult, D.: Photosynthetic activity of intertidal microphytobenthic lík, M., Tušer, M., Bednařík, A., Šimek, K., and Bussmann, I.: Methane dynamics in a large river: a case stud
81, 12, 2019. References q
Migne, A., Gevaert, F., Creach, A., Spilmont, N., Chevalier, E., and Davoult, D.: Photosynthetic activity of intertidal microphytobenthic
communities during emersion: in situ measurements of chlorophyll fluorescence (PAM) and CO2 flux (IRGA), J Phycol, 43, 864-873,
525
10.1111/j.1529-8817.2007.00379.x, 2007. Migne, A., Gevaert, F., Creach, A., Spilmont, N., Chevalier, E., and Davoult, D.: Photosynthetic activity of intertidal microphytobenthic
communities during emersion: in situ measurements of chlorophyll fluorescence (PAM) and CO2 flux (IRGA), J Phycol, 43, 864-873,
525
10.1111/j.1529-8817.2007.00379.x, 2007. Palmer, M., and Ruhi, A.: Measuring Earth's rivers, Science, 361, 546-547, doi:10.1126/science.aau3842, 2018. Paranaíba, J. R., Aben, R., Barros, N., Quadra, G., Linkhorst, A., Amado, A. M., Brothers, S., Catalán, N., Condon, J., Finlayson, C. M.,
Grossart H P Ho itt J Oli eira J nior E S Keller P S Koschorreck M Laas A Leigh C Marcé R Mendonça R M ni
C communities during emersion: in situ measurements of chlorophyll fluorescence (PAM) and CO2 flux (IRGA), J Phycol, 43, 864-873,
525
10.1111/j.1529-8817.2007.00379.x, 2007. Palmer, M., and Ruhi, A.: Measuring Earth's rivers, Science, 361, 546-547, doi:10.1126/science.aau3842, 2018. Paranaíba, J. R., Aben, R., Barros, N., Quadra, G., Linkhorst, A., Amado, A. M., Brothers, S., Catalán, N., Condon, J., Finlayson, C. M.,
Grossart, H.-P., Howitt, J., Oliveira Junior, E. S., Keller, P. S., Koschorreck, M., Laas, A., Leigh, C., Marcé, R., Mendonça, R., Muniz, C. ,
Ruhi, A.: Measuring Earth's rivers, Science, 361, 546-547, doi:10.1126/science.aau3842, 2018. ,
,
,
,
,
, Q
,
,
,
,
,
,
,
,
,
,
,
,
y
,
,
Grossart, H.-P., Howitt, J., Oliveira Junior, E. S., Keller, P. S., Koschorreck, M., Laas, A., Leigh, C., Marcé, R., Mendonça, R., Muniz, C. C., Obrador, B., Onandia, G., Raymundo, D., Reverey, F., Roland, F., Rõõm, E.-I., Sobek, S., von Schiller, D., Wang, H., and Kosten, S.:
530
Cross-continental importance of CH4 emissions from dry inland-waters, Science of The Total Environment, 151925,
https://doi.org/10.1016/j.scitotenv.2021.151925, 2021. C., Obrador, B., Onandia, G., Raymundo, D., Reverey, F., Roland, F., Rõõm, E.-I., Sobek, S., von Schiller, D., Wang, H., and Kosten, S.:
530
Cross-continental importance of CH4 emissions from dry inland-waters, Science of The Total Environment, 151925,
https://doi.org/10.1016/j.scitotenv.2021.151925, 2021. Phillips, C. L., Nickerson, N., Risk, D., and Bond, B. J.: Interpreting diel hysteresis between soil respiration and temperature, Global Change
Biol, 17, 515-527, 10.1111/j.1365-2486.2010.02250.x, 2011. References C., Obrador, B., Onandia, G., Raymundo, D., Reverey, F., Roland, F., Rõõm, E.-I., Sobek, S., von Schiller, D., Wang, H., and Kosten, S.:
530
Cross-continental importance of CH4 emissions from dry inland-waters, Science of The Total Environment, 151925,
https://doi.org/10.1016/j.scitotenv.2021.151925, 2021. p
y
,
,
,
https://doi.org/10.1016/j.scitotenv.2021.151925, 2021. Phillips, C. L., Nickerson, N., Risk, D., and Bond, B. J.: Interpreting diel hysteresis between soil respiration and temperature, Global Change
Biol, 17, 515-527, 10.1111/j.1365-2486.2010.02250.x, 2011. p
g
j
,
Phillips, C. L., Nickerson, N., Risk, D., and Bond, B. J.: Interpreting diel hysteresis between soil respiration and temperature, Global Change
Biol, 17, 515-527, 10.1111/j.1365-2486.2010.02250.x, 2011. Pusch, M., and Fischer, H.: Stoffdynamik und Habitatstruktur in der Elbe, Weißensee, Berlin, 2006. 535
R-Core-Team: R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria, 2016. Raymond, P. A., Zappa, C. J., Butman, D., Bott, T. L., Potter, J., Mulholland, P., Laursen, A. E., McDowell, W. H., and Newbold, D.:
Scaling the gas transfer velocity and hydraulic geometry in streams and small rivers, Limnology and Oceanography-Fluids&Environment,
2, 41-53, 2012. Raymond, P. A., Hartmann, J., Lauerwald, R., Sobek, S., McDonald, C., Hoover, M., Butman, D., Striegl, R., Mayorga, E., Humborg, C.,
540
Kortelainen, P., Durr, H., Meybeck, M., Ciais, P., and Guth, P.: Global carbon dioxide emissions from inland waters, Nature, 503, 355-359,
Doi 10.1038/Nature12760, 2013. R
J A B
A V D
B R H l
M A Li
S S
C M l
k J R
d P A D
C M All
G Raymond, P. A., Hartmann, J., Lauerwald, R., Sobek, S., McDonald, C., Hoover, M., Butman, D., Striegl, R., Mayorga, E., Humborg, C.,
540
Kortelainen, P., Durr, H., Meybeck, M., Ciais, P., and Guth, P.: Global carbon dioxide emissions from inland waters, Nature, 503, 355-359,
Doi 10.1038/Nature12760, 2013. Rosentreter, J. A., Borges, A. V., Deemer, B. R., Holgerson, M. A., Liu, S., Song, C., Melack, J., Raymond, P. A., Duarte, C. M., Allen, G. H., Olefeldt, D., Poulter, B., Battin, T. I., and Eyre, B. D.: Half of global methane emissions come from highly variable aquatic ecosystem
i
5
56
5
545 Doi 10.1038/Nature12760, 2013. Rosentreter, J. A., Borges, A. V., Deemer, B. R., Holgerson, M. A., Liu, S., Song, C., Melack, J., Raymond, P. A., Duarte, C. M., Allen, G. H., Olefeldt, D., Poulter, B., Battin, T. I., and Eyre, B. References H., and Teichert, L.: Temporal patterns and drivers of CO2 emission from dry sediments in agroyne field of a
large river, Biogeosciences, 19, 5221-5236, 10.5194/bg-19-5221-2022, 2022. Liu, S., Kuhn, C., Amatulli, G., Aho, K., Butman, D. E., Allen, G. H., Lin, P., Pan, M., Yamazaki, D., Brinkerhoff, C., Gleason, C., Xia, X., H., and Teichert, L.: Temporal patterns and drivers of CO2 emission from dry sediments in agroyne field of
19, 5221-5236, 10.5194/bg-19-5221-2022, 2022. Liu, S., Kuhn, C., Amatulli, G., Aho, K., Butman, D. E., Allen, G. H., Lin, P., Pan, M., Yamazaki, D., Brinkerhoff, C., Gleason, C., Xia, X.,
and Raymond, P. A.: The importance of hydrology in routing terrestrial carbon to the atmosphere via global streams and rivers, Proceedings
05
of the National Academy of Sciences, 119, e2106322119, doi:10.1073/pnas.2106322119, 2022. and Raymond, P. A.: The importance of hydrology in routing terrestrial carbon to the atmosphere via global streams and rivers, Proceedings
505
of the National Academy of Sciences, 119, e2106322119, doi:10.1073/pnas.2106322119, 2022. Lorke, A., Bodmer, P., Noss, C., Alshboul, Z., Koschorreck, M., Somlai-Haase, C., Bastviken, D., Flury, S., McGinnis, D. F., Maeck, A.,
Mueller, D., and Premke, K.: Technical note: drifting versus anchored flux chambers for measuring greenhouse gas emissions from running
waters, Biogeosciences, 12, 7013-7024, 10.5194/bg-12-7013-2015, 2015. y
p
Lorke, A., Bodmer, P., Noss, C., Alshboul, Z., Koschorreck, M., Somlai-Haase, C., Bastviken, D., Flury, S., McGinnis, D. F., Maeck, A.,
Mueller, D., and Premke, K.: Technical note: drifting versus anchored flux chambers for measuring greenhouse gas emissions from running
waters, Biogeosciences, 12, 7013-7024, 10.5194/bg-12-7013-2015, 2015. ,
g
,
,
,
g
,
Machado dos Santos Pinto, R., Weigelhofer, G., Diaz-Pines, E., Guerreiro Brito, A., Zechmeister-Boltenstern, S., and Hein, T.: River-
510
floodplain restoration and hydrological effects on GHG emissions: Biogeochemical dynamics in the parafluvial zone, Science of The Total
Environment, 715, 136980, https://doi.org/10.1016/j.scitotenv.2020.136980, 2020. g
g
Machado dos Santos Pinto, R., Weigelhofer, G., Diaz-Pines, E., Guerreiro Brito, A., Zechmeister-Boltenstern, S., and Hein, T.: River-
510
floodplain restoration and hydrological effects on GHG emissions: Biogeochemical dynamics in the parafluvial zone, Science of The Total
Environment, 715, 136980, https://doi.org/10.1016/j.scitotenv.2020.136980, 2020. Maeck, A., Hofmann, H., and Lorke, A.: Pumping methane out of aquatic sediments - ebullition forcing mechansims in an impounded river,
biogeosciences, 11, 2925-2938, 2014. sources, Nature Geoscience, 14, 225 230, 10.1038/s41561 021 00715 2, 2021.
545
Sander, R.: Compilation of Henry's law constants (version 4.0) for water as solvent, Atmos. Chem. Phys., 15, 4399-4981, 10.5194/acp-15-
4399-2015, 2015.
Staniek, M.: Spatial distribution of greenhouse gas concentrations along two characteristic Elbe segments, bachelor, Institute of Geoscience
and Geography, MLU, Halle, 2018. 4399-2015, 2015.
Staniek, M.: Spatial distribution of greenhouse gas concentrations along two characteristic Elbe segments, bachelor, Institute of Geoscience
and Geography, MLU, Halle, 2018. References A., Karakaya, N., Barros, N., Howitt,
J. A., Mendoza-Lera, C., Pastor, A., Flaim, G., Aben, R., Riis, T., Arce, M. I., Onandia, G., Paranaíba, J. R., Linkhorst, A., del Campo, R., Amado, A. M., Cauvy-Fraunié, S., Brothers, S., Condon, J., Mendonça, R. F., Reverey, F., Rõõm, E. I., Datry, T., Roland, F., Laas, A.,
Obertegger, U., Park, J. H., Wang, H., Kosten, S., Gómez, R., Feijoó, C., Elosegi, A., Sánchez-Montoya, M. M., Finlayson, C. M., Melita,
495 Obertegger, U., Park, J. H., Wang, H., Kosten, S., Gómez, R., Feijoó, C., Elosegi, A., Sánche
495 Obertegger, U., Park, J. H., Wang, H., Kosten, S., Gómez, R., Feijoó, C., Elosegi, A., Sánchez-Montoya, M. M., Finlayson, C. M., Melita,
495 20 https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. M., Oliveira Junior, E. S., Muniz, C. C., Gómez-Gener, L., Leigh, C., Zhang, Q., and Marcé, R.: Global CO2 emissions from dry inland
waters share common drivers across ecosystems, Nature Communications, 11, art. 2126, https://doi.org/10.1038/s41467-020-15929-y, 2020. Kleinwächter, M., Schröder, U., Rödiger, S., Hentwchel, B., and Anlauf, A.: Buhnen in der Elbe und ihre Umgestaltung, in: Alternative
Buhnenformen in der Elbe - hydraulische und ökologische Wirkungen, Schweizerbart, Stuttgart, 2017. uhnenformen in der Elbe - hydraulische und ökologische Wirkungen, Schweizerbart, Stuttgart, 2017. 500 Koschorreck, M., Prairie, Y. T., Kim, J., and Marce, R.: Technical note: CO2 is not like CH4 - limits of and corrections to the headspace
method to analyse pCO2 in fresh water, Biogeosciences, 18, 1619-1627, 10.5194/bg-18-1619-2021, 2021. Koschorreck, M., Prairie, Y. T., Kim, J., and Marce, R.: Technical note: CO2 is not like CH4 - limits of and corrections to the headspace
method to analyse pCO2 in fresh water, Biogeosciences, 18, 1619-1627, 10.5194/bg-18-1619-2021, 2021. Koschorreck, M., Knorr, K. H., and Teichert, L.: Temporal patterns and drivers of CO2 emission from dry sediments in agroyne field of a
large river, Biogeosciences, 19, 5221-5236, 10.5194/bg-19-5221-2022, 2022. Liu S Kuhn C Amatulli G Aho K Butman D E Allen G H Lin P Pan M Yamazaki D Brinkerhoff C Gleason C Xia X p
method to analyse pCO2 in fresh water, Biogeosciences, 18, 1619-1627, 10.5194/bg-18-1619-2021, 2021. Koschorreck, M., Knorr, K. References D.: Half of global methane emissions come from highly variable aquatic ecosystem
sources Nature Geoscience 14 225 230 10 1038/s41561 021 00715 2 2021
545 Rosentreter, J. A., Borges, A. V., Deemer, B. R., Holgerson, M. A., Liu, S., Song, C., Melack, J., Raymond, P. A., Duarte, C. M., Allen, G. H., Olefeldt, D., Poulter, B., Battin, T. I., and Eyre, B. D.: Half of global methane emissions come from highly variable aquatic ecosystem
sources, Nature Geoscience, 14, 225-230, 10.1038/s41561-021-00715-2, 2021. 545 545 sources, Nature Geoscience, 14, 225-230, 10.1038/s41561-021-00715-2, 2021. 545
Sander, R.: Compilation of Henry's law constants (version 4.0) for water as solvent, Atmos. Chem. Phys., 15, 4399-4981, 10.5194/acp-15-
4399-2015, 2015. Staniek, M.: Spatial distribution of greenhouse gas concentrations along two characteristic Elbe segments, bachelor, Institute of Geoscience
and Geography, MLU, Halle, 2018. ,
,
,
,
,
Sander, R.: Compilation of Henry's law constants (version 4.0) for water as solvent, Atmos. Chem. Phys., 15, 4399-4981, 10.5194/acp-15-
4399-2015, 2015. Staniek, M.: Spatial distribution of greenhouse gas concentrations along two characteristic Elbe segments, bachelor, Institute of Geoscience
and Geography, MLU, Halle, 2018. 21 https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. https://doi.org/10.5194/bg-2023-176
Preprint. Discussion started: 25 September 2023
c⃝Author(s) 2023. CC BY 4.0 License. Stanley, E. H., Casson, N. J., Christel, S. T., Crawford, J. T., Loken, L. C., and Oliver, S. K.: The ecology of methane in streams and rivers:
550
patterns, controls, and global significance, Ecol Monogr, 86, 146-171, 10.1890/15-1027.1, 2016. Striegl, R. G., Dornblaser, M. M., McDonald, C. P., Rover, J. A., and Stets, E. G.: Carbon dioxide and methane emissions from the Yukon
River system, Global Biogeochemical Cycles, 26, https://doi.org/10.1029/2012GB004306, 2012. Tang, M., and Kristensen, E.: Impact of microphytobenthos and macroinfauna on temporal variation of benthic metabolism in shallow coastal Tang, M., and Kristensen, E.: Impact of microphytobenthos and macroinfauna on temporal variation of benthic metabolism in shallow coastal
sediments, J Exp Mar Biol Ecol, 349, 99-112, 10.1016/j.jembe.2007.05.011, 2007. 555 sediments, J Exp Mar Biol Ecol, 349, 99-112, 10.1016/j.jembe.2007.05.011, 2007. 555
UNESCO/IHA: GHG Measurement Guidlines for Freshwater Reservoirs, edited by: Goldenfum, J. A., UNESCO, 138 pp., 2010. Weigold, F., and Baborowski, M.: Consequences of delayed mixing for quality assessment of river water: Example Mulde-Saale-Elbe, J
Hydrol, 369, 296-304, 10.1016/j.jhydrol.2009.02.039, 2009. p
j j
UNESCO/IHA: GHG Measurement Guidlines for Freshwater Reservoirs, edited by: Goldenfum, J. A., UNESCO, 138 pp., 2010. Stanley, E. H., Casson, N. J., Christel, S. T., Crawford, J. T., Loken, L. C., and Oliver, S. K.: The ecology of methane in streams and rivers:
550
patterns, controls, and global significance, Ecol Monogr, 86, 146-171, 10.1890/15-1027.1, 2016.
Striegl, R. G., Dornblaser, M. M., McDonald, C. P., Rover, J. A., and Stets, E. G.: Carbon dioxide and methane emissions from the Yukon
River system, Global Biogeochemical Cycles, 26, https://doi.org/10.1029/2012GB004306, 2012.
Tang M and Kristensen E : Impact of microphytobenthos and macroinfauna on temporal variation of benthic metabolism in shallow coastal References Weigold, F., and Baborowski, M.: Consequences of delayed mixing for quality assessment of river water: Example Mulde-Saale-Elbe, J
Hydrol, 369, 296-304, 10.1016/j.jhydrol.2009.02.039, 2009. Wohl, E., and Iskin, E.: Patterns of Floodplain Spatial Heterogeneity in the Southern Rockies, USA, Geophys Res Lett, 46, 5864-5870,
10.1029/2019gl083140, 2019. 560 22 22
|
https://openalex.org/W2590030572
|
https://link.springer.com/content/pdf/10.1007%2Fs10096-017-2918-7.pdf
|
English
| null |
Serious fungal infections in the Philippines
|
European journal of clinical microbiology & infectious diseases
| 2,017
|
cc-by
| 3,488
|
Eur J Clin Microbiol Infect Dis (2017) 36:937–941
DOI 10.1007/s10096-017-2918-7 Eur J Clin Microbiol Infect Dis (2017) 36:937–941
DOI 10.1007/s10096-017-2918-7 ORIGINAL ARTICLE Serious fungal infections in the Philippines M. C. R. Batac1 & D. Denning2 Received: 21 December 2016 /Accepted: 21 December 2016 /Published online: 21 February 2017
# The Author(s) 2017. This article is published with open access at Springerlink.com Received: 21 December 2016 /Accepted: 21 December 2016 /Published online: 21 February 2017
# The Author(s) 2017. This article is published with open access at Springerlink.com 391, oral candidiasis 3,467, esophageal candidiasis 1,522
(all in HIV-infected people), invasive aspergillosis (IA)
3,085, candidemia 1,968, candida peritonitis 246,
mucormycosis 20, fungal keratitis 358, tinea capitis 846 and
mycetoma 97 annually. A total of 1,852,137 (1.9% of popu-
lation) are afflicted with a serious fungal infection. Epidemiological studies are needed to validate these esti-
mates, facilitating appropriate medical care of patients and
proper prioritization of limited resources. Abstract The Philippines is a low middle-income, tropical
country in Southeast Asia. Infectious diseases remain the main
causes of morbidity, including tuberculosis. AIDS/HIV prev-
alence is still low at <1%, but is rapidly increasing. Fungal
disease surveillance has not been done, and its burden has
never been estimated. This becomes more important as the
population of immunocompromised patients increases, drawn
from patients with AIDS, TB, malignancies, and autoimmune
diseases requiring chronic steroid use. Using the methodology
of the LIFE program (www.LIFE-worldwide.org), estimates
were derived from data gathered from WHO, UNAIDS,
Philippine Health Statistics 2011, Philippine Dermatological
Society Health Information System database, HIV/AIDS and
ART registry of the Philippines, epidemiological studies such
as The TREAT Asia HIV Observational Database 2005, and
personal communication. Aspergillosis and candidiasis were
the top causes of fungal infections in the Philippines. Chronic
pulmonary aspergillosis (CPA), drawn from the number of
tuberculosis patients, affects 77,172 people. Allergic
bronchopulmonary aspergillosis (ABPA) and severe asthma
with fungal sensitization (SAFS) frequencies, which were de-
rived from the number of asthmatic patients, affect 121,113
and 159,869 respectively. Recurrent vulvovaginal candidiasis
(RVVC) affects 1,481,899 women. Other estimates
were cryptococcal meningitis 84, Pneumocystis pneumonia 1
University of Manchester, Manchester, UK
2
Education and Research Center, University Hospital of South
Manchester, Southmoor Road, Manchester M23 9LT, UK Methods Applying the methodology of the LIFE program (www.LIFE-
worldwide.org), prevalence and incidence of serious fungal
infections were derived from data gathered from WHO,
UNAIDS, Philippine Health Statistics 2011, Philippine
Dermatological Society (PDS) Health Information System da-
tabase, HIV/AIDS and ART registry of the Philippines, epi-
demiological studies such as The TREAT Asia HIV
Observational Database 2005, and personal communication. HIV/AIDS cases in the Philippines are few with a cumu-
lative number of cases at 34,999 since the first case was
reported in 1984. However, a sudden rise in cases has been
observed since 2009, with 83% of cumulative cases diag-
nosed between 2009 to present. Most of the newly diag-
nosed cases are in the 25–29 age group, and among men
who have sex with men [4, 8]. Our estimated incidences of
Pneumocystis pneumonia, cryptococcal meningitis, and oral
candidiasis were based on HIV/AIDS cases in the recent
past, which were relatively low but are increasing rapidly. Adequate diagnosis of opportunistic fungal disease with the
highly sensitive and specific cryptococccal antigen lateral
flow device [9, 10] and Pneumocystis PCR are required
[11, 12]. It is not known if disseminated histoplasmosis con-
tributes to illness and death in AIDS in the Philippines, but
it is likely. Histoplasmin reactivity has been documented,
and at least nine cases of histoplasmosis have been reported
[2, 13, 14]. Hand in hand with an effective AIDS prevention
program and prompt and adequate treatment of cases with
antiretroviral drugs, proper management of opportunistic
fungal infections is required. Introduction In the Philippines, most of the initial epidemiological studies
on mycoses focused on superficial fungal infections [1–3]. Since the 1950s, superficial mycoses have been in the top five
most common diagnoses in outpatient dermatology clinics [2]. For several decades, fungal diagnostic procedures in the
Philippines were not routinely done and were limited to the
isolation, culture, and identification of fungi involved in skin,
hair, and nail infections. Two university-based microbiology
departments conduct short courses in Diagnostic Mycology
on a regular basis. In the past, these used to cater solely for
dermatologists, because the course was a requirement in their
residency training. However, with the advent of AIDS and the
compromised patient, more infectious disease fellows and
medical laboratory personnel have enrolled in these courses. * D. Denning
ddenning@manchester.ac.uk
M. C. R. Batac
tinbatacmd@gmail.com The advent of AIDS, first described in the Philippines in
1984, led to a greater recognition of fatal fungal diseases [4]. Out of 470 HIV/AIDS patients seen in a tertiary general hos-
pital in the Philippines, 32.5% presented with opportunistic
infections [5]. Among the opportunistic infections, 32% were
due to Pneumocystis pneumonia, 4% to cryptococcal menin-
gitis, and 3% to oral thrush [5]. HIV cases are climbing steeply
in the Philippines (20–30% annual increase in the years 2013– Eur J Clin Microbiol Infect Dis (2017) 36:937–941 938 Table 1
Important population statistics Table 1
Important population statistics
Percenta
Number
Population (2013)
98,394,000
Males
50.5
49,688,970
Females
49.5
48,705,030
Below 15 years old
34.0
33,453,960
Above 40 years old
25.6
25,188,864
Above 60 years old
6.0
5,903,640
Females 15–50 years old
25.9
25,484,046
a (Epidemiology Bureau, Department of Health 2011) 2016), especially among men [4]. Patients who are on immu-
nosuppressive agents, or are immunocompromised due to
medical conditions and/or are critically ill, are also at an in-
creased risk of acquiring serious fungal infections, and such
patients are common in the urban centres of the Philippines. There is no active surveillance done for serious fungal infec-
tions in the country, and epidemiological studies even for be-
nign superficial mycoses have been sparse. Although nation-
ally based surveillance programs are the gold-standard in es-
timating disease prevalence, they are usually costly and diffi-
cult to implement [6]. Introduction Therefore, the actuarial method used by
previous researchers was used in this study to estimate the
incidence and prevalence of serious fungal diseases, in which
results from previous epidemiological studies, populations at
risk, and epidemiological databases were used [6]. a (Epidemiology Bureau, Department of Health 2011) RVVC per month. There are around 4,000 to 5,000 obstetri-
cians in the Philippines, so the maximum estimate for the
prevalence of RVVC would only be 240,000, ∼1.2 million
less than the estimate. Those with RVVC who consult a phy-
sician do not represent everyone in the population, and many
women suffer in silence. It is likely that many patients self-
medicate, since patients may purchase topical antifungals
without prescription or try local remedies. Clearly, this sub-
stantial issue needs addressing in the Philippines. RVVC per month. There are around 4,000 to 5,000 obstetri-
cians in the Philippines, so the maximum estimate for the
prevalence of RVVC would only be 240,000, ∼1.2 million
less than the estimate. Those with RVVC who consult a phy-
sician do not represent everyone in the population, and many
women suffer in silence. It is likely that many patients self-
medicate, since patients may purchase topical antifungals
without prescription or try local remedies. Clearly, this sub-
stantial issue needs addressing in the Philippines. Population and country profile The Philippines is classified as lower middle income by the
World Bank. It is considered one of the promising newly in-
dustrialized countries, enjoying a steady economic growth at
the turn of the millennium, with a per capita 2013 gross do-
mestic product of $2,765. Based on the 2013 WHO statistical
profile, the population of the Philippines is 98,394,000, with
34% under the age of 15, and 6% over the age of 60. The
median age is 23. The number of hospital beds per population
was the lowest in Asia in 2010 [7]. Table 1 shows relevant
population statistics. The Philippine prevalence of IA, CPA, ABPA, and
SAFS, diseases caused by Aspergillus sp., has never been
studied. However, several cases of pulmonary aspergillo-
sis have been documented. In 1975, a case of pulmonary
aspergillosis was reported in Manila in a 25-year-old male
who presented with a 7-year history of hemoptysis with-
out fever and anorexia, and who was diagnosed with tu-
berculosis but did not respond to anti-tuberculosis medi-
cines [15]. From February 1982 to May 1990, 16 cases of
pulmonary aspergillosis were seen at the Lung Center of Burden of fungal infections In 2016, a total of 1,852,137 severe fungal infections are esti-
mated to have occurred in the Philippines. Table 2 shows the
incidence and prevalence for selected serious fungal infec-
tions. Eighty percent of this is due to RVVC. However, local
practicing obstetricians think this number is too high, and
approximate that each general obstetrician sees 12–15 patients
with vulvovaginal candidiasis, three to four of whom have Eur J Clin Microbiol Infect Dis (2017) 36:937–941 939 Table 2
Prevalence and incidence of selected serious fungal infections in the Philippines, 2016 Table 2
Prevalence and incidence of selected serious fungal infections in the Philippines, 2016
Serious fungal iInfection
Estimation method
Totals
Rate/100,000
Cryptococcal meningitis
7% of new AIDS diagnosis
84
Annual incidence
0.09
Pneumocystis pneumonia
31% of new AIDS diagnosis
391
Annual incidence
0.40
Invasive aspergillosis (IA)
10% of AML + an equal number in other
haematological conditions, 0.5% of
renal transplant patients, 4% of liver
transplant patients, and 1.3% of patients
hospitalized for COPD
3,085
Annual incidence
3
Chronic pulmonary aspergillosis (CPA)
22% of cavitary pulmonary TB, 2% of
non-cavitary pulmonary TB, annually,
reduced by 15% for surgery and death. Burden of fungal infections 77,172
Prevalence
78
Allergic bronchopulmonary
aspergillosis (ABPA)
2.5% of adult asthmatics
121,113
Prevalence
123
Severe asthma with fungal
sensitisation (SAFS)
33% of the most severe 10% of adult
asthmatics
159,869
Prevalence
162
Candidaemia
2/100,000 general population, 1.5 in
non-ICU and 0.5 in ICU
1,968
Annual incidence
2
Candida peritonitis
50% of incidence of candidemia in ICU
246
Annual incidence
0.25
Oral candidiasis
90% of HIV patients not on antiretrovirals
and CD4 < 200 × 106/L
3,467
Annual incidence
3.5
Oesophageal candidiasis
20% of HIV patients not on antiretrovirals and
CD4 < 200 × 106/l + 5% of HIV patients on
antiretrovirals
1,522
Annual incidence
1.5
Recurrent vulvovaginal candidiasis
(RVVC) (>4x/year)
5% of women 15–50 years of age
1,481,899
Prevalence
3,012
Mucormycosis
0.2 cases per 1,000,000 population
20
Annual incidence
0.02
Fungal keratitis
Based on cases seen at a tertiary government
hospital in the NCR in 2015
358
Annual incidence
0.36
Tinea capitis
Based on cases seen at Philippine Dermatological
Society training institutions in 2015
846
Annual incidence
0.86
Mycetoma
Based on cases seen at Philippine
Dermatological Society training
institutions in 2015
97
Annual incidence
0.10
Total serious fungal infection burden
1,852,137 Candidemia among infants confined in the neonatal inten-
sive care unit was reported to affect 56 of 7,830 infants in
2004, and 31 of 7,830 infants in 2005 [21]. A case–control
study done in the same facility revealed that birthweight of
1,250 to 2,499 grams and gestational age of <28 weeks were
significantly associated with candidemia (p < 0.05) [21]. The
rate of candidemia in different countries varied from 1.2 to 25
per 100,000 population [22]. We have decided to use two per
100,000 population since exposure to risk factors for
candidemia may be low, and there are remarkably few hospi-
tals for this populous country [7]. In some developed coun-
tries, candidemia rates are increasing instead of decreasing
and may be due to increased survival of patients with severe
diseases or extremely low-birth weight, more aggressive in-
terventions in the form of surgery, transplants, invasive proce-
dures and devices, immunosuppressive therapy, and use of
broad-spectrum antibiotics [22]. These are resources that are
not accessible to most Filipinos. the Philippines [16]. Nine of them had inactive pulmonary
tuberculosis; 13 underwent lobectomy or segmentectomy
or cyst removal. From Jan 2000 to December 2002, 37
patients were diagnosed with aspergilloma, and all were
surgically managed. References Both were caused by Fonsecaea compacta. 6. Beardsley J, Denning DW, Chau NV, Yen NTB, Crump JA, Day JN
(2015) Estimating the burden of fungal disease in Vietnam. Mycoses 58:101–106. doi:10.1111/myc.12382 7. OECD WHO (2014) Hospital care. In: Health at a Glance: Asia/
Pacific 2014. Measuring Progress towards Universal Health
Coverage. OECD Publishing, Paris, pp 66–68 8. Ross AGP, Ditangco RA, Belimac JG, Olveda RM, Mercado ES,
Rogers GD, Cripps AW, Lam A, Crowe SM (2013) HIV epidemic
in men who have sex with men in the Philippines. Lancet Infect Dis
13:472–473. doi:10.1016/S1473-3099(13)70129-4 9. WHO (2011) Rapid advice: diagnosis, prevention and management
of cryptococcal disease in HIV-infected adults, adolescents and
children. World Health Organization, Geneva 10. Tang MW, Clemons K V, Katzenstein DA, Stevens DA (2015) The
cryptococcal antigen lateral flow assay: A point-of-care diagnostic
at an opportune time. Crit Rev Microbiol 1–9 11. Calderón EJ, Gutiérrez-Rivero S, Durand-Joly I, Dei-Cas E (2010)
Pneumocystis infection in humans: diagnosis and treatment. Expert
Rev Anti Infect Ther 8:683–701. doi:10.1586/eri.10.42 12. Global Action Fund for Fungal Infections (2014) GAFFI Roadmap:
improving outcomes for patients with fungal infections across the
world - a road map for the next decade. GAFFI, Geneva 13. Bulmer AC, Bulmer GS (2001) Incidence of histoplasmin hyper-
sensitivity in the Philippines. Mycopathologia 149:69–71. doi:10.1023/A:1007277602576 14. Tacastacas J, Bacorro E, Berba RP, Tobias-Altura M, Sison MA,
Roa F (2006) A 47 year old male with multiple ulcerated skin
plaques: a case report on disseminated histoplasmosis and a review
of eight previously reported cases of histoplasmosis in the
Philippines. Philipp J Microbiol Infect Dis 35:5–11 Challenges in the management of serious fungal infections
in a resource-limited setting include timely diagnosis, proper
antifungal intervention, patient compliance with long-term
treatment, and high cost of antifungal treatment. 15. Alora A, Alora B, Dizon G (1975) A report of a case of
pulmonary aspergillosis in the Philippines. Philipp J Microbiol
Infect Dis 4:46–52 Acknowledgements
We would like to acknowledge the help given by
Dr. Vanessa de Villa (Founding Member of the Philippine Society for
Transplant Surgeons and Chair of the Philippine Board of Transplant
Surgery), Dr. Richard Nepomuceno (cataract, cornea, refractive surgery),
Dr. Edsel Maurice T. Salvana (infectious diseases and tropical medicine,
HIV/AIDS), Dr. Ricardo Manalastas (obstetrics and gynecology, infec-
tious diseases) through Dr. Ma. Cristina Pelaez-Crisologo, Dr. Francisco
Lopez (bone marrow transplant specialist) through Dr. References case was detected. Back then, experts surmised that tinea
capitis was uncommon in the Philippines and was not a public
health problem, and this seems the case to this day [2]. The
common dermatophytes isolated were Microsporum canis,
Trichophyton tonsurans, T. mentagrophytes, and
M. gypseum [2]. A total of 9,180 (11.5% of new patients)
consulted for superficial mycoses in ten PDS institutions in
2015. Fungal keratitis, tinea capitis, mycetoma, and RVVC
are mycoses that affect young, productive, and otherwise
healthy patients. These affect work productivity and quality
of life. It is important to manage these diseases promptly to
avoid complications. 1. Handog EB, Dayrit JF (2005) Mycology in the Philippines,
revisited. Jpn J Med Mycol 46:71–76. doi:10.3314/jjmm.46.71 2. Simuangco S, Bocobo F, Lacuna L (1962) Review of literature on
medical mycology in the Philippines, 1955-1962. Mycopathol
Mycol Appl 20:145–156. doi:10.1080/00150198808201399 3. Simuangco S (1975) Geographic dermatology: the Philippines. Acta Med Philipp 11:37–43 4. NHSS (2016) HIV/AIDS and ART registry of the Philippines
(HARP). Manila 5. Salvana EM, Leyritana K, Alejandria M, Lim J, Destura R, Tenorio
A (2012) Opportunistic infections in Filipino HIV patients. Abstract published and presented at the ID Week 2012
Conference on October 2012 in San Diego, CA.. The incidence of subcutaneous mycoses is rare in the
Philippines, but these usually cause disfiguring lesions in
young people, limiting their mobility and affecting quality of
life [1]. By 1963, three patients with eumycetoma caused
by Penicillium funiculosum, Madurella grisea, and
Scedosporium (formerly Monosporium) apiospermum had
been seen in the Philippines [23]. Eumycetoma of 20 years
duration in a 40-year-old man was found to be caused by
Phialemonium sp., and was treated with terbinafine 250 mg
twice a day for 9 months followed by excision of redundant
skin [24]. Another case was reported in 2009 of a 33-year-old
man with draining sinuses and nodules on the left foot and
thigh, from which Madurella sp. was isolated [25]. More re-
cently, Madurella mycetomatis was cultured from the foot
lesions of 9 years duration of a 33-year-old man [26]. Most
of these patients were farmers. If eumycetoma presents with
chronic nodules and draining sinuses, chromoblastomycosis
appears as warty lesions. A few cases of chromoblastomycosis
have also been reported in the Philippines, including a 58-
year-old man who presented with verrucous lesions of 25years
and a 71-year-old man with warty lesions of 48 years duration
[2, 27]. Burden of fungal infections Ninety-two percent presented with
hemoptysis and 95% had tuberculosis [17]. In a retrospec-
tive study done in a private hospital in Manila, there
were 14 cases of aspergillosis seen from 1998-2004; five
were diabetics and three had renal transplants. Eleven
presented with pulmonary aspergillosis, and three had
extrapulmonary aspergillosis [18]. A case of primary la-
ryngeal aspergillosis in a 28-year-old who presented with
fever, cough, and cold a day after giving birth by caesar-
ean section was reported [19]. The Philippines is endemic
for TB, and urban and rural areas have high rates of
COPD [20]. TB and COPD are both associated with
CPA, and therefore it is probable that there are many
undiagnosed CPA cases, with some of them managed as
resistant TB cases. It is important to determine baseline
prevalence using available serological tests. the Philippines [16]. Nine of them had inactive pulmonary
tuberculosis; 13 underwent lobectomy or segmentectomy
or cyst removal. From Jan 2000 to December 2002, 37
patients were diagnosed with aspergilloma, and all were
surgically managed. Ninety-two percent presented with
hemoptysis and 95% had tuberculosis [17]. In a retrospec-
tive study done in a private hospital in Manila, there
were 14 cases of aspergillosis seen from 1998-2004; five
were diabetics and three had renal transplants. Eleven
presented with pulmonary aspergillosis, and three had
extrapulmonary aspergillosis [18]. A case of primary la-
ryngeal aspergillosis in a 28-year-old who presented with
fever, cough, and cold a day after giving birth by caesar-
ean section was reported [19]. The Philippines is endemic
for TB, and urban and rural areas have high rates of
COPD [20]. TB and COPD are both associated with
CPA, and therefore it is probable that there are many
undiagnosed CPA cases, with some of them managed as
resistant TB cases. It is important to determine baseline
prevalence using available serological tests. In 1955, dermatologists surveyed the schools where two
children with tinea capitis studied; however, no additional Eur J Clin Microbiol Infect Dis (2017) 36:937–941 940 References References Charina Dizon,
and the Philippine Dermatological Society — Health Information
Systems. 16. Ramos A (1991) Pulmonary aspergillosis nine-year experience at
Lung Center of the Philippines. Sci Proc 75–82 17. Castolo JP (2003) Experience in surgery for pulmonary
aspergilloma. Philipp J Thorac Cardiovasc Surg 10:6–9 18. Llorin R, Chua J (2005) Aspergillosis: a case series. Int J
Antimicrob Agents 26S:2005 19. Villanueva JC, Opulencia AP, Calavera KZ, Lim W (2015)
Primary laryngeal aspergillosis in a postpartum patient. 30:
47–49 20. Idolor LF, De Guia TS, Francisco NA, Roa CC, Ayuyao FG, Tady
CZ, Tan DT, Banal-Yang S, Balanag VM, Reyes MTN, Dantes RB
(2011) Burden of obstructive lung disease in a rural setting in the
Philippines. Respirology 16:1111–1118. doi:10.1111/j.1440-
1843.2011.02027.x Open Access This article is distributed under the terms of the Creative
Commons Attribution 4.0 International License (http://
creativecommons.org/licenses/by/4.0/), which permits unrestricted use,
distribution, and reproduction in any medium, provided you give appro-
priate credit to the original author(s) and the source, provide a link to the
Creative Commons license, and indicate if changes were made. 21. Morales-Lagunzad NRM, Mantaring JBV (2010) Risk factors for
candidemia in the neonatal intensive care unit of the Philippine Eur J Clin Microbiol Infect Dis (2017) 36:937–941 941 General Hospital from October 2003 To August 2006 : a case-
control Study. Pediatr Infect Dis Soc Philipp J 10:44–50 25. Dela Cerna EM, Montinola F (2009) Eumycetoma in a Filipino
adult man. J Philipp Dermatol Soc 18:41–43 22. Arendrup MC (2010) Epidemiology of invasive candidiasis. Curr
Opin Crit Care 16:445–452. doi:10.1097/MCC.0b013e32833e84d2 26. Pena-Dumdum A, Banate-Gulfan G, Ledesma TG, Gabriel MT,
Senador L (2016) Stage III eumycetoma successfully treated with oral
ketoconazole and surgical debulking. J Philipp Dermatol Soc 25:58–61 23. Reyes A (1963) A contribution to the study of mycetoma in
the Philippines: maduromycosis caused by monosporium
apiospermum (laboratory studies). Acta Med Philipp 19:89–
102 27. Alora MBT, Bulmer GS (1993) Chromomycosis : report of a case
successfully treated with itraconazole. Philipp J Microbiol Infect Dis
22:23–27 28. Epidemiology Bureau Department of Health (2011) The 2011
Philippine Health Statistics. Manila 24. Abad-Venida ML, Reyes MV (2001) Eumycetoma (Phialemonium
spp) cured by terbinafine. J Philipp Soc Cutan Med 8–11 28. Epidemiology Bureau Department of Health (2011) The 2011
Philippine Health Statistics. Manila
|
https://openalex.org/W3165519204
|
https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0252208&type=printable
|
English
| null |
Social capital and cognitive decline: Does sleep duration mediate the association?
|
PloS one
| 2,021
|
cc-by
| 7,576
|
PLOS ONE RESEARCH ARTICLE Background Citation: Wang L, Li J, Wang Z, Du Y, Sun T, Na L,
et al. (2021) Social capital and cognitive decline:
Does sleep duration mediate the association? PLoS
ONE 16(5): e0252208. https://doi.org/10.1371/
journal.pone.0252208 Studies have found that social capital (SC) is associated with the risk of cognitive decline;
however, the mechanism explaining how SC leads to cognitive decline is unclear. The cur-
rent study examines the mediation effect of sleep duration on the relationship between SC
and cognitive decline in Chinese older adults. Editor: Y Zhan, German Centre for
Neurodegenerative Diseases Site Munich:
Deutsches Zentrum fur Neurodegenerative
Erkrankungen Standort Munchen, GERMANY
Received: September 7, 2020
Accepted: May 12, 2021
Published: May 27, 2021 Editor: Y Zhan, German Centre for
Neurodegenerative Diseases Site Munich:
Deutsches Zentrum fur Neurodegenerative
Erkrankungen Standort Munchen, GERMANY Liqun Wang1, Jiangping Li1, Zhizhong WangID2*, Yong Du3,4, Ting Sun3, Li Na3,
Yang Niu5,6* 1 Department of Epidemiology and Statistics, School of Public Health and Management at Ningxia Medical
University, Yinchuan, China, 2 Department of Epidemiology and Statistics at School of Public Health of
Guangdong Medical University, Dongguan, China, 3 Surgical Laboratory of General Hospital, Ningxia
Medical University, Yinchuan, China, 4 School of Clinical Medicine at Ningxia Medical University, Yinchuan,
China, 5 Key Laboratory of the Ningxia Ethnomedicine Modernization, Ministry of Education, Ningxia Medical
University, Yinchuan, China, 6 School of Traditional Chinese Medicine, Ningxia Medical University,
Yinchuan, China a1111111111
a1111111111
a1111111111
a1111111111
a1111111111 a1111111111
a1111111111
a1111111111
a1111111111
a1111111111 * wzhzh_lion@126.com (ZW); niuyang0227@163.com (YN) Results Peer Review History: PLOS recognizes the
benefits of transparency in the peer review
process; therefore, we enable the publication of
all of the content of peer review and author
responses alongside final, published articles. The
editorial history of this article is available here:
https://doi.org/10.1371/journal.pone.0252208 After controlling for covariates, both social cohesion and social interaction were positively
correlated with the MMSE score (p<0.001), and social cohesion was negatively correlated
with sleep duration (p = 0.009); On the contrary, sleep duration was negatively correlated
with MMSE score (p<0.001). Linear regression analysis showed social cohesion was posi-
tively associated with the MMSE score (β = 0.16, p = 0.005), while sleep duration was asso-
ciated with an increased risk of cognitive decline (β = -0.72, p<0.001). Sleep duration has
mediated the relationship between social cohesion and cognitive decline (explaining 21.7%
of the total variance). Copyright: © 2021 Wang et al. This is an open
access article distributed under the terms of the
Creative Commons Attribution License, which
permits unrestricted use, distribution, and
reproduction in any medium, provided the original
author and source are credited. Methods A cross-sectional study of 955 community-dwelling aged 60 or over was conducted. The
mini-mental state examination (MMSE), self-report sleep duration questionnaire, and social
capital scales were administered during the face-to-face survey. The Bootstrap methods
PROCESS program is employed to test the mediation model. Received: September 7, 2020
Accepted: May 12, 2021
Published: May 27, 2021 Received: September 7, 2020
Accepted: May 12, 2021
Published: May 27, 2021 Social capital and cognitive decline: Does
sleep duration mediate the association? Liqun Wang1, Jiangping Li1, Zhizhong WangID2*, Yong Du3,4, Ting Sun3, Li Na3,
Yang Niu5,6* PLOS ONE PLOS ONE * wzhzh_lion@126.com (ZW); niuyang0227@163.com (YN) Introduction Expanded longevity is one of the most remarkable success stories in human history, and this
also directs the population aging. The proportion of people aged 60 years and older is expected
to rise to 22% of the total population in the coming decades, which is from 800 million to 2 bil-
lion [1]. One of the consequences of rapid population aging is the increased prevalence of
aging-related diseases, of which dementia and mild cognitive impairment were the commonly
prevalent neuropsychiatric disorders in older adults [2]. The determinants of cognitive decline include biological (Apolipoprotein E, protein tau, β-
amyloid) and environmental factors (education, lifestyle, diet, medical service, social capital,
etc.) [3–5]. Studies have also reported that sleep duration is an independent neurobehavioral
predictor of cognitive disorders [6]. A meta-analysis suggested that longer sleep duration was
associated with higher risks of mild cognitive impairment [7]. Another study revealed a U-
shaped association that showed either shorter or longer sleep duration was associated with a
higher risk of cognitive decline [8]. Social capital (SC) is a characteristic of social life, including interpersonal trust, norms of
reciprocity, mutual aid, and social involvement (like socializing with friends, relatives, col-
leagues, or neighbors) [9], which has linked with several beneficial health outcomes [10, 11]. As a social determinant of health, SC may play an essential role in protecting individuals from
cognitive decline. At least one study has found that SC accrued in early and midlife may reduce
the detrimental influences of psychological stress on cognitive functioning in later life [12]. Specifically, possessing a rich SC (supportive personal network with numerous types of rela-
tionships, e.g., neighbor and friends) has been associated with better cognitive function among
the old adults [13–17]. One possible explanation is individuals with rich SC lead to less life
stress and more leisure time [12]. Also, one study found that poor SC might increase the risk
of cognitive decline among elderly residents in Wuhan, China [18]. Besides, SC has been associated with sleep duration [19, 20]. A notable exception is one
study that revealed an inverted U-shaped association between SC and sleep duration [19]. Takahashi et al. [21] reported that Japanese workers who had higher neighborhood or work-
place social capital had a better quantity of sleep (not too short and too long). There still unclear how SC leads to better cognitive function in older individuals. Conclusions Data Availability Statement: All relevant data are
within the paper and its Supporting Information
files. Social capital negatively associated with the risk of cognitive decline in this Chinese popula-
tion, and sleep duration may partly explain this relationship. It may be a suggestive clue to 1 / 13 PLOS ONE | https://doi.org/10.1371/journal.pone.0252208
May 27, 2021 PLOS ONE Social capital and cognitive decline identify those at a higher risk of progressing to cognitive impairment. Further prospective
study in need to confirm this finding due to the cross-sectional design. Funding: The study was supported by the
Research and Development Plan of the 13th five-
year plan of Ningxia autonomous region (the major
S&T projects.) (grant number 2016BZ02) and the
National Natural Science Foundation of China
(grant number 81860599). Competing interests: The authors have declared
that no competing interests exist. Introduction A previous
study reported that shorter sleep duration mediated the association between homocysteine
and cognitive decline [22]. They argued that short sleep duration might cause an increased
homocysteine level, then strengthened Aβ accumulation, which is a critical pathological pro-
cess of cognitive decline. The present study sought to examine the mediating effect of sleep
duration on the relationship between SC and cognitive decline in old Chinese adults. We
hypothesized that rich SC is associated with better cognitive function and that this association
would be mediated at least in part by appropriate sleep duration. The field process The trained medical students served as investigators. With local community leaders’ coopera-
tion, the investigators visited the participants’ houses to guide them to finish the survey, then
described our study and questionnaire. Under the participants’ agreement, our investigators
read the questions one by one to them and then recorded their answers, and the survey lasted
approximately 45 minutes. As for respondents with low mini-mental state examination
(MMSE) scores, other information is provided by family members. The finished questionnaire
was double-checked immediately by a separate supervisor in the field. Study sample Data were abstracted from a cross-sectional survey conducted from April 2017 to July 2017 at
Ningxia province, China. The detailed sampling process can be found elsewhere [23]. Here, in 2 / 13 PLOS ONE | https://doi.org/10.1371/journal.pone.0252208
May 27, 2021 PLOS ONE Social capital and cognitive decline summary, the participants were selected using a multi-stage sampling method: firstly, four
counties were selected from a total of 22 counties in the province using a stratified sampling
design according to the proportion of the minority population and the economic status. Sec-
ondly, twenty rural communities and twenty urban communities were selected among the
four selected counties (with a total of 166 urban communities and 628 rural communities)
using random sampling methods. Thirdly, 115 households were selected in each target com-
munities using a systematic sampling method. Finally, one eligible family member from each
household was determined to attend the survey using the Kish table. There were 615 house-
holds not responded after three times attempt to contact, and 3,985 eligible participants were
finally selected. Of them, 1159 participants were aged 60 and over, and 104 participants were
excluded due to the cognitive function test missing. Of them, 1159 participants were aged 60
and over, and 104 participants were excluded due to the cognitive function test’s missing
value. Ultimately, 955 participants were included in this study (Fig 1). The inclusion criteria
are a) living at the present address for at least six months and b) aged 60 years or older. The
exclusion criteria were the following: a) unconsciousness caused by any forms; b) the acute
phase of a cerebrovascular accident; c) a severe illness (e.g., stroke, cerebral infarction or
myocardial infarction) that prevents communication; d) any obvious cognitive disabilities or
deafness, aphasia or other language barriers; e) people reported with depression (by asking “do
you ever be told have depression by your doctors”) and f) with sleep disorders and taking hyp-
notics or psychotropic medications, as well as some particular occupation need to going to bed
late. Fig 1. Participant screen process. https://doi.org/10.1371/journal.pone.0252208.g001 3 / 13 PLOS ONE | https://doi.org/10.1371/journal.pone.0252208
May 27, 2021 PLOS ONE Social capital and cognitive decline The Institutional Review Board of the General Hospital of Ningxia Medical University
(approval number 2017–200) approved this study. All the participants provided a written con-
sent form at the beginning of the survey. PLOS ONE | https://doi.org/10.1371/journal.pone.0252208
May 27, 2021 Measurement Cognitive function. The Chinese version of the MMSE scale was employed to assess the
cognitive function, which has high sensitivity (90.8%) and specificity (93%) for screening cog-
nitive disorders [24]. The MMSE consists of 19 questions to measure the five different domains
of cognitive function (1) orientation, (2) memory, (3) attention and calculation, (4) language,
and (5) constructional praxis (coping task, e.g. copying intersecting pentagons). And has a
total score ranging from 0 to 30; a higher score reflects better cognitive function. We categori-
fied cognitive performance into two categories (cognitive decline vs. normal according to Cui
et al. suggested criteria [25]: MMSE 17 for those with no formal education; MMSE 20 for
those with primary school education (6 years); and MMSE24 for those with junior high
school education or above ( 9 years). Social capital. SC was evaluated using the social capital scale to cover the two dimensions
of social capital (social cohesion and social interaction) developed by Mujahid [26]. The social
cohesion subscale consists of 4 items: 1) People around here are willing to help their neighbors,
2) People in my neighborhood generally get along with each other, 3) People in my neighbor-
hood can be trusted, 4) People in my neighborhood share the same values. Each item ranged
from 1 to 5 (1 = strongly disagree, 2 = disagree, 3 = neutral, 4 = agree, and 5 = strongly agree). Cronbach’s alpha was 0.88 among the Chinese sample [27], and among this sample was 0.81. The social interaction scale consists of five items: 1) You and other people in the community
(village) help each other (e.g., look after children, help buy something and borrow tools), 2)
When a neighbor is not at home or going out, you can help him look after the house or prop-
erty, 3) people in the community (village) talk about each other’s personal matters (children
care, exercise, etc.), 4) participate in group activities together with people in the community
(village), and 5) communicate with each other on the street. Each item scored from 1 to 4 in
response to a 4-point Likert scale (from never to often). The previous study has shown the Chi-
nese version of the social interaction scale has good reliability and validity [27]. The Cron-
bach’s alpha in this sample was 0.76. Social capital. Demographic characteristics of participants The demographic characteristics of the participants were shown in Table 1. The average age
was 66.9 (SD = 5.3) years, with a range of 60 to 80 years. Slightly about half (46.7%) were male,
the mean educational attainment was 3.7 (SD = 4.2) years, and about 81.9% were married. The
mean sleep duration was 8.1 (SD = 1.5) hours, the mean score of social cohesion was 15.7
(SD = 2.5), and for the social interaction was 13.2 (SD = 3.7). The mean score of MMSE was
23.6 (SD = 5.4). And the prevalence of cognitive decline (CD) was 15.8%. Participants with CD
were older, more likely to have longer sleep duration, and lower SC scores than those with nor-
mal cognitive performance. Statistical analyses Analyses were performed using the Statistical Package for the Social Sciences (SPSS) version
24.0 (IBM Inc., Chicago, Illinois, USA). We performed a logarithmic transformation of the
MMSE score to fit the normal distribution. Means and standard deviations (SD) or were used
to describe continuous variables; counts and proportions were used to describe categorical var-
iables. The bivariate test using Student’s t-test or Chi-square test. Partial correlations were
employed to create the correlation matrix under controlling the socio-demographic covariates. Three separate linear regression models were performed to examine the association among
SC, sleep duration, and MMSE. In summary, model l include the SC and sleep duration;
model 2 adjusted with covariate variables (age, gender, ethnicity, residence, educational attain-
ment, marital status, family income, body mass index, smoking, alcohol use, fasting blood glu-
cose, hyperlipidemia, hypertension); and the interaction between SC and sleep duration were
added in model 3. And collinearity detected where the tolerance >0.1 and Variance Inflation
Factor (VIF) <5 indicate multicollinearity can be ignored [28]. Bootstrap methods of PRO-
CESS developed by Hayes was employed to test the mediation effect of sleep duration on the
relationship between SC and MMSE score [29]. The bias-corrected percentile bootstrap confi-
dence interval does not contain 0 indicate that the mediation effect statistically significant [30]. Sensitivity analyses were performed using Structural Equation Modelling (SEM) approach. Measurement SC was evaluated using the social capital scale to cover the two dimensions
of social capital (social cohesion and social interaction) developed by Mujahid [26]. The social
cohesion subscale consists of 4 items: 1) People around here are willing to help their neighbors,
2) People in my neighborhood generally get along with each other, 3) People in my neighbor-
hood can be trusted, 4) People in my neighborhood share the same values. Each item ranged
from 1 to 5 (1 = strongly disagree, 2 = disagree, 3 = neutral, 4 = agree, and 5 = strongly agree). Cronbach’s alpha was 0.88 among the Chinese sample [27], and among this sample was 0.81. Sleep duration. Three questions “What time do you usually go to sleep at night?”, “What
time do you usually rise in the morning?” and “In general, do you take afternoon nap often?”. The time between bedtime and rise up as the crude sleep duration, then it adjusted depends on
the response to the question “In general, do you take afternoon nap often?” if the answer is yes,
then the modified sleep duration is crude sleep duration plus one hour. Socio-demographic information includes age, gender (male vs. female), ethnicity (Han vs. minority), residence (rural vs. urban), educational attainment (continuous data), marital status
(married vs. unmarried/widowed/divorced), family income (as measured by the self-reported 4 / 13 PLOS ONE | https://doi.org/10.1371/journal.pone.0252208
May 27, 2021 PLOS ONE Social capital and cognitive decline family average individual income per month and the answer included five groups: <1,000
RMB, 1,000–1,999 RMB, 2,000–2,999 RMB, 3,000–4,999 RMB, and 5,000 RMB or more) were
collected using a standard form. The data about body mass index (BMI = weight (kg)/height (m)2), fasting blood glucose
(mmol/L), smoking (defined as at least one cigarette per day and last for six months or more),
alcohol use (defined as a drink of at least one glass of alcohol, that equals 1/2 bottle of beer or
125-milliliter grape wine or fruit wine or 40-milliliter white wine, in a day for the past 12
months), hypertension (yes vs. no), dyslipidemia (yes vs. no) were abstracted from the medical
record. PLOS ONE Social capital and cognitive decline Table 1. Demographic characteristics of participants (n = 955). Variables
Total (N = 955)
cognitive decline (N = 151)
normal (N = 804)
χ2/t
p
Age, mean (SD), years
66.9(5.3)
68.7(5.8)
66.6(5.1)
4.08a
<0.001
Gender, male, n (%)
446(46.7)
36(23.8)
410(51.0)
37.66 b
<0.001
Ethnicity, han, n (%)
646(67.6)
109(72.2)
537(66.8)
2.83 b
0.243
Residence, rural, n (%)
479(50.2)
88(58.3)
391(48.6)
4.73 b
0.030
Marital status, n (%)
3.10 b
0.078
unmarried/widowed/divorced
173(18.1)
35(23.2)
138(17.2)
Married
782(81.9)
116(76.8)
666(82.8)
Educational attainment, mean (SD), years
3.7(4.2)
1.6(3.7)
4.0(4.1)
7.27 a
<0.001
Family income, n (%)
37.95 b
<0.001
<1000 RMB
505(52.9)
111(73.5)
394(49.0)
1000~1999 RMB
192(20.1)
17(11.2)
175(21.7)
2000~2999 RMB
148(15.5)
14(9.3)
134(16.7)
3000~4999 RMB
87(9.1)
6(4.0)
81(10.1)
5000 RMB
23(2.4)
3(2.0)
20(2.5)
Smoking, n (%)
181(19.0)
18(11.9)
163(20.3)
5.77 b
0.016
Alcohol use, n (%)
126(13.2)
5(3.3)
121(15.0)
15.29 b
<0.001
BMI, mean (SD)
25.8(7.9)
25.5(4.0)
25.8(8.3)
0.44 a
0.662
FBG, mean (SD)
5.1(1.6)
5.0(1.4)
5.1(1.6)
0.53a
0.599
Hyperlipidemia, n (%)
38(4.0)
3(2.0)
35(4.4)
1.86 b
0.172
Hypertension, n (%)
385(40.3)
60(39.7)
325(40.4)
0.02 b
0.874
Social cohension, mean (SD)
15.7(2.5)
15.3(2.8)
15.7(2.5)
1.89 a
0.059
Social interaction, mean (SD)
13.2(3.7)
12.5(4.3)
13.3(3.6)
2.57 a
0.010
Sleep duration, mean (SD), hours
8.1(1.5)
9.0(1.6)
7.9(1.4)
7.58 a
<0.001 -0.11, P = 0.003). As shown in Fig 2, a U-shaped association between sleep duration and
MMSE score was found. SD: standard deviation; BMI: body mass index; FBG: fasting blood glucose
a: t-test used
b: chi-square test used. https://doi.org/10.1371/journal.pone.0252208.t001 -0.11, P = 0.003). As shown in Fig 2, a U-shaped association between sleep duration and
MMSE score was found. -0.11, P = 0.003). As shown in Fig 2, a U-shaped association between sleep duration and
MMSE score was found. The binary correlation matrix The partial correlation matrix showed in Table 2. After controlling for socio-demographic var-
iables (age, gender, ethnicity, residence, educational attainment, marital status, family income)
and health variables (BMI, smoking, alcohol use, FBG, hyperlipidemia, hypertension), both
the social cohesion (r = 0.12, P<0.001) and social interaction (r = 0.09, P<0.05) were positively
correlated with MMSE score; the sleep duration was negatively correlated with MMSE score (r
= -0.24, P<0.001); and the social cohesion was negatively correlated with sleep duration (r = 5 / 13 PLOS ONE | https://doi.org/10.1371/journal.pone.0252208
May 27, 2021 SD: standard deviation; BMI: body mass index; FBG: fasting blood glucose Linear regression analysis duration was inversely associated with the MMSE score (model 2). The same was true for
social interaction. The association between SC and MMSE score disappeared when adding the
interaction between SC and sleep duration (model 3), indicating that sleep duration is a possi-
ble mediator. Model l + covariate variables (age, gender, ethnicity, residence, educational attainment, marital status, family income, body Model l = SC+ sleep duration; Model 2 = Model l + covariate variables (age, gender, ethnicity, residence, educational attainment, marital status, family income, body
mass index, smoking, alcohol use, fasting blood glucose, hyperlipidemia, hypertension); Model 3 = Model 2 + interaction between SC and sleep duration; Social
interaction and social cohesion were tested separately in all the models. β: beta; 95%CI: 95% confidence interval; NA: not applicable.
R2 for social cohesion in model 1–3 are 0.081, 0.389, 0.389 respectively; R2 for social interaction in model 1–3 are 0.089, 0.388, 0.389 respectively. were tested separately in all the models. β: beta; 95%CI: 95% confidence interval; NA: not applicable. e, fasting blood glucose, hyperlipidemia, hypertension); Model 3 = Model 2 + interaction between SC and sleep duration; Soc p
y
β
;
;
pp
n in model 1–3 are 0.081, 0.389, 0.389 respectively; R2 for social interaction in model 1–3 are 0.089, 0.388, 0.389 respectively. https://doi.org/10.1371/journal.pone.0252208.t003 mass index, smoking, alcohol use, fasting blood glucose, hyperlipidemia, hypertension); Model 3 = Model 2 + interaction between SC and sleep duration; Social
interaction and social cohesion were tested separately in all the models. β: beta; 95%CI: 95% confidence interval; NA: not applicable.
R2 for social cohesion in model 1 3 are 0 081 0 389 0 389 respectively; R2 for social interaction in model 1 3 are 0 089 0 388 0 389 respectively Linear regression analysis Linear regression analysis
As Table 3 showed, social cohesion was positively associated with the MMSE score; namely,
those with rich social cohesion may predict better cognitive performance. In contrast, sleep
Table 2. Correlation matrix between SC, sleep duration, and MMSE score (n = 955)a. Mean
SD
1
2
3
4
1.MMSE
23.6
5.4
1
2.Social cohension
15.7
2.5
0.12
1
3.Social interaction
13.2
3.7
0.09
0.46
1
4.Sleep duration
8.1
1.5
-0.24
-0.11
-0.05
1
p<0.001
p<0.05, SD = standard deviation
a: The covariates include age, gender, ethnicity, residence, educational attainment, marital status, family income, BMI, smoking, alcohol use, FBG, hyperlipidemia,
hypertension were adjusted using Partial correlation method. https://doi.org/10.1371/journal.pone.0252208.t002 As Table 3 showed, social cohesion was positively associated with the MMSE score; namely,
those with rich social cohesion may predict better cognitive performance. In contrast, sleep Table 2. Correlation matrix between SC, sleep duration, and MMSE score (n = 955)a. Mean
SD
1
2
3
4
1.MMSE
23.6
5.4
1
2.Social cohension
15.7
2.5
0.12
1
3.Social interaction
13.2
3.7
0.09
0.46
1
4.Sleep duration
8.1
1.5
-0.24
-0.11
-0.05
1
p<0.001
p<0.05, SD = standard deviation
a: The covariates include age, gender, ethnicity, residence, educational attainment, marital status, family income, BMI, smoking, alcohol use, FBG, hyperlipidemia,
hypertension were adjusted using Partial correlation method Table 2. Correlation matrix between SC, sleep duration, and MMSE score (n = 955)a. Mean
SD
1
2
3
4
1.MMSE
23.6
5.4
1
2.Social cohension
15.7
2.5
0.12
1
3.Social interaction
13.2
3.7
0.09
0.46
1
4.Sleep duration
8.1
1.5
-0.24
-0.11
-0.05
1 Table 2. Correlation matrix between SC, sleep duration, and MMSE score (n = 955)a. p<0.05, SD = standard deviation
a: The covariates include age, gender, ethnicity, residence, educational attainment, marital status, family income, BMI, smoking, alcohol use, FBG, hyperlipidemia,
hypertension were adjusted using Partial correlation method. PLOS ONE | https://doi.org/10.1371/journal.pone.0252208
May 27, 2021 6 / 13 PLOS ONE Social capital and cognitive decline Fig 2. A U-shaped association between sleep duration and MMSE score. https://doi.org/10.1371/journal.pone.0252208.g002 Fig 2. A U-shaped association between sleep duration and MMSE score. https://doi.org/10.1371/journal.pone.0252208.g002 duration was inversely associated with the MMSE score (model 2). The same was true for
social interaction. The association between SC and MMSE score disappeared when adding the
interaction between SC and sleep duration (model 3), indicating that sleep duration is a possi-
ble mediator. Mediation effect of sleep duration on the relationship of SC and cognitive
decline As shown in Table 4, after controlling for covariates, there is a significant mediation effect of
sleep duration on the relationship between social cohesion and cognitive decline. The results
showed that both the direct effect (p = 0.004) and the indirect effect (p = 0.002) were significant. The mediation effect explained 21.7% (0.045/0.207) of the total variance. No mediation effect of
sleep duration in the relationship between social interaction and cognitive decline was found. Table 3. Linear regression model for interaction between SC and sleep duration on cognitive decline (n = 955). Variables
Model 1
Model 2
Model 3
P value
β (95%CI)
P value
β (95%CI)
P value
β (95%CI)
Social cohesion
0.192
0.09(-0.04,0.22)
0.005
0.16(0.05,0.27)
0.522
0.23(-0.39,0.84)
Sleep duration
<0.001
-0.99(-1.21,-0.77)
<0.001
-0.72(-0.91,-0.52)
0.311
-0.60(-1.75,0.55)
Social cohesion×sleep duration
NA
NA
NA
NA
0.900
-0.01(-0.08,0.07)
Social interaction
0.001
0.14(0.06, 0.23)
0.020
0.09(0.01,0.16)
0.607
0.11(-0.31,0.52)
Sleep duration
<0.001
-0.98(-1.20,-0.76)
<0.001
-0.74(-0.93,-0.54)
0.037
-0.71(-1.37,-0.05)
Social interaction×sleep duration
NA
NA
NA
NA
0.950
-0.01(-0.05,0.05) Table 3. Linear regression model for interaction between SC and sleep duration on cognitive decline (n = 955). Variables
Model 1
Model 2
Model 3
P value
β (95%CI)
P value
β (95%CI)
P value
β (95%CI)
Social cohesion
0.192
0.09(-0.04,0.22)
0.005
0.16(0.05,0.27)
0.522
0.23(-0.39,0.84)
Sleep duration
<0.001
-0.99(-1.21,-0.77)
<0.001
-0.72(-0.91,-0.52)
0.311
-0.60(-1.75,0.55)
Social cohesion×sleep duration
NA
NA
NA
NA
0.900
-0.01(-0.08,0.07)
Social interaction
0.001
0.14(0.06, 0.23)
0.020
0.09(0.01,0.16)
0.607
0.11(-0.31,0.52)
Sleep duration
<0.001
-0.98(-1.20,-0.76)
<0.001
-0.74(-0.93,-0.54)
0.037
-0.71(-1.37,-0.05)
Social interaction×sleep duration
NA
NA
NA
NA
0.950
-0.01(-0.05,0.05)
Model l = SC+ sleep duration; Model 2 = Model l + covariate variables (age, gender, ethnicity, residence, educational attainment, marital status, family income, body
mass index, smoking, alcohol use, fasting blood glucose, hyperlipidemia, hypertension); Model 3 = Model 2 + interaction between SC and sleep duration; Social
interaction and social cohesion were tested separately in all the models. β: beta; 95%CI: 95% confidence interval; NA: not applicable. R2 for social cohesion in model 1–3 are 0.081, 0.389, 0.389 respectively; R2 for social interaction in model 1–3 are 0.089, 0.388, 0.389 respectively. https://doi.org/10.1371/journal.pone.0252208.t003 able 3. Linear regression model for interaction between SC and sleep duration on cognitive decline (n = 955). Table 3. Sensitivity analysis As showed in Fig 3, the SEM approach was peformed to sensitivity analysis. The results showed
social cohension was positively associated with cognitive function and inversely related to sleep
duration, and sleep duration negatively associated with cognitive function. These results was
consistent with the binary correlation results even though the pathway coefficient was not the
same due to the different methods. This validated the methodological robustness of findings. PLOS ONE Social capital and cognitive decline Table 4. The mediating effect of sleep duration on the relationship between SC and cognitive decline . Effect
Bias-Corrected 95%CI
β
SE
P-value
Lower
Upper
Social cohesion
Total effect
0.207
0.058
<0.001
0.093
0.332
Indirect Effects
0.045
0.016
0.002
0.018
0.082
Direct Effects
0.162
0.057
0.004
0.050
0.275
Social interaction
Total effect
0.104
0.039
0.007
0.026
0.181
Indirect Effects
0.004
0.002
0.133
-0.001
0.009
Direct Effects
0.090
0.038
0.019
0.015
0.165
After controlling for age, gender, ethnicity, residence, educational attainment, marital status, family income, BMI, smoking, alcohol use, FBG, hyperlipidemia,
hypertension. After controlling for age, gender, ethnicity, residence, educational attainment, marital status, family income, BMI, smoking
hypertension. After controlling for age, gender, ethnicity, residence, educational attainment, marital status, family income, BMI, smoking, alcohol use, FBG, hyperlipidemia,
hypertension. Considering the possible U-shape relationship between sleep duration and cognitive
decline, the exploring analysis conducted stratified (use a cutoff point of mean sleep duration
of the total sample) by sleep duration showed in Table 5. The mediation effect of sleep dura-
tion on the relationship between social cohesion and cognitive decline persists in those who
had eight hours and longer sleep duration. However, the mediation effect disappears in those
who had less than eight hours of sleep duration per day. Mediation effect of sleep duration on the relationship of SC and cognitive
decline Linear regression model for interaction between SC and sleep duration on cognitive dec Model l = SC+ sleep duration; Model 2 = Model l + covariate variables (age, gender, ethnicity, residence, educational attainment, marital status, family income, body
mass index, smoking, alcohol use, fasting blood glucose, hyperlipidemia, hypertension); Model 3 = Model 2 + interaction between SC and sleep duration; Social
interaction and social cohesion were tested separately in all the models. β: beta; 95%CI: 95% confidence interval; NA: not applicable. R2 for social cohesion in model 1–3 are 0.081, 0.389, 0.389 respectively; R2 for social interaction in model 1–3 are 0.089, 0.388, 0.389 respectively. PLOS ONE | https://doi.org/10.1371/journal.pone.0252208
May 27, 2021 7 / 13 a: The effect adjusted the covariates include age, gender, ethnicity, residence, educational attainment, marital status, family income, BMI, smoking, alcohol use, FBG,
hyperlipidemia, hypertension. https://doi.org/10.1371/journal.pone.0252208.t005 Discussion The current study examined the association between SC and cognitive decline. And provides
the primary evidence for the relationship between SC and cognitive decline among older Table 5. The mediation model of social cohesion stratified by sleep durationa. Effect
Bias-Corrected 95%CI
β
SE
P-value
Lower
Upper
sleep duration <8 h
Total effect
0.196
0.093
0.038
0.011
0.380
Indirect Effects
-0.001
0.008
0.954
-0.012
0.022
Direct Effects
0.197
0.093
0.038
0.011
0.380
sleep duration 8 h
Total effect
0.164
0.074
0.026
0.019
0.309
Indirect Effects
0.041
0.021
0.025
0.008
0.091
Direct Effects
0.123
0.072
0.089
-0.019
0.265
a: The effect adjusted the covariates include age, gender, ethnicity, residence, educational attainment, marital status, family income, BMI, smoking, alcohol use, FBG,
h
l
d
h Table 5. The mediation model of social cohesion stratified by sleep durationa. PLOS ONE | https://doi.org/10.1371/journal.pone.0252208
May 27, 2021 PLOS ONE | https://doi.org/10.1371/journal.pone.0252208
May 27, 2021 8 / 13 PLOS ONE Social capital and cognitive decline Fig 3. https://doi.org/10.1371/journal.pone.0252208.g003 https://doi.org/10.1371/journal.pone.0252208.g003 adults, suggesting a possible mechanism to explain how SC is related to reduce cognitive
impairment in studies examining that association. As hypothesized, the results revealed a posi-
tive association of SC with cognitive performance in older Chinese adults, and this relationship
was partially mediated by sleep duration. The mediation effect is accounting for 21.7% of the
total effect in the total sample, while no mediation effect was found among those who had less
than eight hours of sleep duration. Sleep duration was negatively associated with MMSE. Long sleep duration might be a risk
factor for cognitive decline, consistent with prior research that found long sleep duration was
associated with cognitive impairment [31–33]. A cohort study found long sleep duration was
associated with poorer cognitive performance among adults aged 41–75 years [34]. Previous
studies also have reported that long sleep duration is associated with low MMSE scores among
old adults [35–39]. One possible explanation is increased sleep fragmentation associated with
decreased cognitive performance, and so it may be that longer sleep duration may emerge
more frequent nighttime wakes or sleep in bed much more time [40]. Additionally, obstructive
sleep apnea syndrome (OSAS) can cause deterioration in cognitive functions, and the study
reported that OSAS was more prevalent in extended sleep duration groups [41, 42]. PLOS ONE | https://doi.org/10.1371/journal.pone.0252208
May 27, 2021 Strength and limitations Given the increasing rate of aging and the incidence of dementia in the elderly Chinese popu-
lation, the present findings have relevance for understanding the mechanisms of how SC is
linked with cognitive function. And provide primary evidence for developing interventional
program for cognitive decline in minority areas. Several limitations were identified; first, the
cross-sectional design prevents making causal inferences from the relationships between SC
and cognitive decline reported here. Hence, further longitudinal design would be necessary to
determine causal relationships in the future. Second, potential confounding variables like
depression that were not included in the analysis may lead to overestimation of the association
between cognitive function and sleep duration. Third, due to the feasibility consideration, bed-
time and sleep duration were collected via a self-reported survey question; it may involve infor-
mation bias even though it has been found to have a reasonable correlation with actigraphic
measurement [56]. Discussion Besides,
long sleep duration has been associated with an increased level of inflammatory factors [43,
44], and elevated inflammatory cytokine levels increase the risk of cognitive decline [45]. Fur-
thermore, reports revealed a U-shaped association with a higher risk of cognitive decline in
older adults with either short or long sleep duration [7, 46, 47]. Compared with lower SC, the average sleep duration was shorter in those who have higher
SC. This finding was consistent with the previous study that found lower neighborhood SC
was negatively associated with short sleep duration among Japanese male adults [17]. China is
a typical Guanxi-based society, and the previous study has reported that Guanxi (traditional
Chinese social interaction) has almost the same connotation as social capital [48]. Addition-
ally, rural China’s culture values trust, mutual assistance, and reciprocal exchange, which pro-
vide cultural soil for cultivating social capital [49]. The current study also found SC was positively correlated with the MMSE score and might
be a protective factor of cognitive function. One study conducted in Taiwan revealed that
increased social support is associated with better cognitive function in older adults [50]. Also,
Holtzman and his colleagues reported a positive association between social support and 9 / 13 PLOS ONE | https://doi.org/10.1371/journal.pone.0252208
May 27, 2021 PLOS ONE Social capital and cognitive decline cognitive function among older adults [51]. Moreover, a three-year cohort study found that
social networks reduced the incidence of dementia in older adults [52]. On the one hand, the
possible mechanisms were social activities provide the challenge of effective communication
and participation in complex interpersonal exchanges [53]. On the other hand, emotional sup-
port might buffer against physiological stress and benefit cognitive function [54]. Furthermore,
a recent study manifested that social capital could help older adults continue to independently
live in local communities and handle life stressors efficiently, even when they encounter
declines in their physical and cognitive health. Social capital can provide older residents with a
sense of security and belonging and is an important reserve domain in old age [55]. Conclusions Social cohesion, one dimension of the social capital, positively associated with cognitive func-
tion, and sleep duration partly mediated this relationship. The findings provides the primary
evidence for better understanding how SC is related to reduce risk of cognitive decline in stud-
ies examining that association. Hence, clinicians can suggest patients communicate more with
others (chat, play chess or exercise together, etc.) to improve the social capital, and in turn
maintain their cognitive function. References 1. Bloom DE, Chatterji S, Kowal P, Lloyd-Sherlock P, McKee M, Rechel B, et al. Macroeconomic implica-
tions of population ageing and selected policy responses. Lancet. 2015; 385: 649–657. https://doi.org/
10.1016/S0140-6736(14)61464-1 PMID: 25468167 2. World Health Organization. Dementia: a public health priority. http://www.who.int/mental_health/
publications/demeatia_report_2012/en/ 3. Wang LQ, Wang ZZ, Koenig HG, Alshohaib S. Interactions between Apolipoprotein E Genes and Religi-
osity in Relation to Mild Cognitive Impairment. Neuropsychiatry. 2017; 07: 659–666. https://doi.org/10. 4172/Neuropsychiatry.1000262 4. Buerger K, Teipel SJ, Zinkowski R, Blennow K, Arai H, Engel R, et al. CSF tau protein phosphorylated
at threonine 231 correlates with cognitive decline in MCI subjects. Neurology. 2002; 59: 627–629. https://doi.org/10.1212/wnl.59.4.627 PMID: 12196665 5. Li X, Ma C, Zhang J, Liang Y, Chen Y, Chen K, et al. Prevalence of and potential risk factors for mild
cognitive impairment in community-dwelling residents of Beijing. J Am Geriatr Soc. 2013; 61: 2111–
2119. https://doi.org/10.1111/jgs.12552 PMID: 24479143 6. Chen JC, Espeland MA, Brunner RL, Lovato LC, Wallace RB, Leng X, et al. Sleep duration, cognitive
decline, and dementia risk in older women. Alzheimers Dement. 2016; 12: 21–33. https://doi.org/10. 1016/j.jalz.2015.03.004 PMID: 26086180 7. Liang Y, Qu LB, Liu H. Non-linear associations between sleep duration and the risks of mild cognitive
impairment/dementia and cognitive decline: a dose-response meta-analysis of observational studies. Aging Clin Exp Res. 2019; 31: 309–320. https://doi.org/10.1007/s40520-018-1005-y PMID: 30039452 8. Wu L, Sun DL, Tan Y. A systematic review and dose-response meta-analysis of sleep duration and the
occurrence of cognitive disorders. Sleep Breath. 2018; 22: 805–814. https://doi.org/10.1007/s11325-
017-1527-0 PMID: 28589251 9. Kawachi I, Kim D, Coutts A, Subramanian SV. Commentary: Reconciling the three accounts of social
capital. Int J Epidemiol. 2004; 33: 682–690. https://doi.org/10.1093/ije/dyh177 PMID: 15282222 10. Verhaeghe PP and Tampubolon G. Individual social capital, neighbourhood deprivation, and self-rated
health in England. social science & medicine. 2012; 75: 349–357. https://doi.org/10.1016/j.socscimed. 2012.02.057 PMID: 22560798 11. Ziersch AM. Health implications of access to social capital: findings from an Australian study. Social Sci-
ence & Medicine. 2005; 61: 2119–2131. https://doi.org/10.1016/j.socscimed.2005.01.015 PMID:
16115715 12. Ihle A, Oris M, Sauter J, Rimmele U, Kliegel M. Cognitive Reserve and Social Capital Accrued in Early
and Midlife Moderate the Relation of Psychological Stress to Cognitive Performance in Old Age. Dement Geriatr Cogn Disord. 2018; 45: 190–197. https://doi.org/10.1159/000488052 PMID: 29870984 13. Bennett DA, Schneider JA, Tang Y, Arnold SE, Wilson RS. Author Contributions Conceptualization: Liqun Wang, Zhizhong Wang, Li Na, Yang Niu. Conceptualization: Liqun Wang, Zhizhong Wang, Li Na, Yang Niu. Data curation: Liqun Wang, Jiangping Li, Zhizhong Wang, Yong Du, Ting Sun. Formal analysis: Liqun Wang, Jiangping Li, Zhizhong Wang. Supervision: Zhizhong Wang, Yang Niu. Writing – original draft: Liqun Wang, Zhizhong Wang. Writing – review & editing: Jiangping Li, Zhizhong Wang. 10 / 13 PLOS ONE | https://doi.org/10.1371/journal.pone.0252208
May 27, 2021 PLOS ONE Social capital and cognitive decline PLOS ONE | https://doi.org/10.1371/journal.pone.0252208
May 27, 2021 References The effect of social networks on the relation
between Alzheimer’s disease pathology and level of cognitive function in old people: a longitudinal
cohort study. Lancet Neurol. 2006; 5: 406–412. https://doi.org/10.1016/S1474-4422(06)70417-3
PMID: 16632311 14. Crooks VC, Lubben J, Petitti DB, Little D, Chiu V. Social network, cognitive function, and dementia inci-
dence among elderly women. Am J Public Health. 2008; 98: 1221–1227. https://doi.org/10.2105/AJPH. 2007.115923 PMID: 18511731 15. Ellwardt L, Van Tilburg TG, Aartsen MJ. The mix matters: Complex personal networks relate to higher
cognitive functioning in old age. Soc Sci Med. 2015; 125: 107–115. https://doi.org/10.1016/j. socscimed.2014.05.007 PMID: 24840784 16. Kimura D, Takeda T, Ohura T, Imai A. Evaluation of facilitative factors for preventing cognitive decline:
A 3-year cohort study of community intervention. Psychogeriatrics. 2016; 17: 9–16. https://doi.org/10. 1111/psyg.12182 PMID: 26858148 17. Wang B, He P, Dong B. Associations between social networks, social contacts, and cognitive function
among Chinese nonagenarians/centenarians. Arch Gerontol Geriatr. 2015; 60: 522–527. https://doi. org/10.1016/j.archger.2015.01.002 PMID: 25650253 18. Wang C, Zhu J, Cai Y, Cui D, Wang Q, Mao Z. Community-Based Study of the Relationship Between
Social Capital and Cognitive Function in Wuhan, China. Asia Pac J Public Health. 2016; 28: 717–724. https://doi.org/10.1177/1010539516640351 PMID: 27029384 19. Win T, Yamazaki T, Kanda K, Tajima K, Sokejima S. Neighborhood social capital and sleep duration: a
population based cross-sectional study in a rural Japanese town. BMC Public Health. 2018; 18: 343. https://doi.org/10.1186/s12889-018-5204-4 PMID: 29529998 11 / 13 PLOS ONE | https://doi.org/10.1371/journal.pone.0252208
May 27, 2021 PLOS ONE Social capital and cognitive decline 20. Cheng GH, Chan A, Lo JC. Importance of social relationships in the association between sleep duration
and cognitive function: data from community-dwelling older Singaporeans. Int Psychogeriatr. 2018; 30:
893–901. https://doi.org/10.1017/S1041610217001041 PMID: 28615083 21. Takahashi M, Tsutsumi A, Kurioka S, Inoue A, Shimazu A, Kosugi Y, Kawakami N. Occupational and
socioeconomic differences in actigraphically measured sleep. J Sleep Res. 2014; 23: 458–462. https://
doi.org/10.1111/jsr.12136 PMID: 24628714 22. Sanchez-Espinosa MP, Atienza M, Cantero JL. Sleep mediates the association between homocysteine
and oxidative status in mild cognitive impairment. Sci Rep. 2017; 7: 7719. https://doi.org/10.1038/
s41598-017-08292-4 PMID: 28798397 23. Wang L, Li J, Du Y, Sun T, Na L, Wang Z. The relationship between sleep onset time and cardiometa-
bolic biomarkers in Chinese communities: a cross-sectional study. BMC Public Health. 2020; 20:374. https://doi.org/10.1186/s12889-020-08516-9 PMID: 32197597 24. Zhang ZX, Zahner GE, Roma´n GC, Liu XH, Wu CB, Hong Z, et al. References Socio-demographic variation of
dementia subtypes in china: Methodology and results of a prevalence study in Beijing, Chengdu,
Shanghai, and Xian. Neuroepidemiology. 2006; 27: 177–187. https://doi.org/10.1159/000096131
PMID: 17035714 25. Cui GH, Yao YH, Xu RF, Tang HD, Jiang GX, Wang Y, et al. Cognitive impairment using education-
based cutoff points for CMMSE scores in elderly Chinese people of agricultural and rural Shanghai
China. Acta Neurol Scand. 2011; 124: 361–367. https://doi.org/10.1111/j.1600-0404.2010.01484.x
PMID: 21303351 26. Mujahid MS, Diez Roux AV, Morenoff JD, Raghunathan T. Assessing the measurement properties of
neighborhood scales: from psychometrics to ecometrics. Am J Epidemiol. 2007; 165: 858–867. https://
doi.org/10.1093/aje/kwm040 PMID: 17329713 27. Gao J, Fu H, Li J, Jia Y. Association between social and built environments and leisure-time physical
activity among Chinese older adults: a multilevel analysis. BMC Public Health. 2015; 15:1–11. https://
doi.org/10.1186/1471-2458-15-1 PMID: 25563658 28. Mason Charlotte H, Perreault William D. Collinearity, Power, and Interpretation of Multiple Regression
Analysis. Journal of Marketing Research. 1991; 28: 268–280. 29. Hayes A. Introduction to mediation, moderation, and conditional process analysis. Journal of Educa-
tional Measurement. 2013; 51: 335–337. 30. MacKinnon DP, Fritz MS, Williams J, Lockwood CM. Distribution of the product confidence limits for the
indirect effect: Program PRODCLIN. Behav Res Methods. 2007; 39: 384–389. https://doi.org/10.3758/
bf03193007 PMID: 17958149 31. Schmutte T, Harris S, Levin R, Zweig R, Katz M, Lipton R. The relation between cognitive functioning
and self-reported sleep complaints in nondemented older adults: results from the Bronx aging study. Behav Sleep Med. 2007; 5: 39–56. https://doi.org/10.1207/s15402010bsm0501_3 PMID: 17313323 32. Loerbroks A, Debling D, Amelang M, Stu¨rmer T. Nocturnal sleep duration and cognitive impairment in a
population-based study of older adults. Int J Geriatr Psychiatry. 2010; 25: 100–109. https://doi.org/10. 1002/gps.2305 PMID: 19548221 33. Malek-Ahmadi M, Kora K, O’Connor K, Schofield S, Coon D, Nieri W. Longer self-reported sleep dura-
tion is associated with decreased performance on the montreal cognitive assessment in older adults. Aging Clin Exp Res. 2015; 28: 333–337. https://doi.org/10.1007/s40520-015-0388-2 PMID: 26063636 34. van Oostrom SH, Nooyens ACJ, van Boxtel MPJ, Verschuren WMM. Long sleep duration is associated
with lower cognitive function among middle-age adults-the Doetinchem Cohort Study. Sleep Med. 2018; 41: 78–85. https://doi.org/10.1016/j.sleep.2017.07.029 PMID: 29425581 35. Ramos AR, Dong C, Elkind MS, Boden-Albala B, Sacco RL, undek T, et al. Association between sleep
duration and the mini-mental score: the Northern Manhattan study. J Clin Sleep Med. 2013; 9: 669–
673. https://doi.org/10.5664/jcsm.2834 PMID: 23853560 36. References Auyeung TW, Lee JS, Leung J, Kwok T, Leung PC, Woo J, et al. Cognitive deficit is associated with
phase advance of sleep-wake rhythm, daily napping, and prolonged sleep duration-a cross-sectional
study in 2,947 community-dwelling older adults. Age. 2013; 35: 479–486. https://doi.org/10.1007/
s11357-011-9366-6 PMID: 22215376 37. Virta JJ, Heikkila¨ K, Perola M, Koskenvuo M, Ra¨iha¨ I, Rinne JO, et al. Midlife sleep characteristics asso-
ciated with late life cognitive function. Sleep. 2013; 36: 1533–1541. https://doi.org/10.5665/sleep.3052
PMID: 24082313 38. Benito-Leo´n J, Louis ED, Bermejo-Pareja F. Cognitive decline in short and long sleepers: A prospective
population-based study (NEDICES). J Psychiatr Res. 2013; 47: 1998–2003. https://doi.org/10.1016/j. jpsychires.2013.09.007 PMID: 24094933 12 / 13 PLOS ONE | https://doi.org/10.1371/journal.pone.0252208
May 27, 2021 PLOS ONE Social capital and cognitive decline 39. Faubel R, Lo´pez-Garcı´a E, Guallar-Castillo´n P, Graciani A, Banegas J R, Rodrı´guez-Artalejo F. Usual
sleep duration and cognitive function in older adults in Spain. J Sleep Res. 2010; 18: 427–435. https://
doi.org/10.1111/j.1365-2869.2009.00759.x 40. Blackwell T, Yaffe K, Ancoli-Israel S, Redline S, Ensrud KE, Stefanick ML, et al. association of sleep
characteristics and cognition in older community-dwelling men: the MrOS sleep study. Sleep. 2011; 34:
1347–1356. https://doi.org/10.5665/SLEEP.1276 PMID: 21966066 41. Bliwise DL, King AC, Harris RB. Habitual sleep durations and health in a 50–65 year old population. J
Clin Epidemiol. 1994; 47: 35–41. https://doi.org/10.1016/0895-4356(94)90031-0 PMID: 8283193 42. Be´dard MA, Montplaisir J, Richer F, Rouleau I, Malo J. Obstructive Sleep Apnea Syndrome: Pathogen-
esis of Neuropsychological Deficits. J Clin Exp Neuropsychol. 1991; 13: 950–964. https://doi.org/10. 1080/01688639108405110 PMID: 1779033 43. Grandner MA, Buxton OM, Jackson N, Sands-Lincoln m, Pandey A, Jean-Louis G. Extreme sleep dura-
tions and increased C-reactive protein: effects of sex and ethnoracial group. Sleep. 2013; 36:769–779. https://doi.org/10.5665/sleep.2646 PMID: 23633760 44. Patel SR, Zhu X, Storfer-Isser A, Mehra R, Jenny NS, Tracy R, et al. Sleep duration and biomarkers of
inflammation. Sleep. 2009; 32:200–204. https://doi.org/10.1093/sleep/32.2.200 PMID: 19238807 45. Windham BG, Simpson BN, Lirette S, Bridges J, Bielak L, Peyser PA, et al. Associations Between
Inflammation and Cognitive Function in African Americans and European Americans. Journal of the
American Geriatrics Society. 2014; 62: 2303–2310. https://doi.org/10.1111/jgs.13165 PMID: 25516026 46. Xu L, Jiang CQ, Lam TH, Liu B, Jin YL, Zhu T, et al. Short or long sleep duration is associated with mem-
ory impairment in older Chinese: the Guangzhou Biobank Cohort Study. Sleep. 2011; 34: 575–580. https://doi.org/10.1093/sleep/34.5.575 PMID: 21532950 47. PLOS ONE | https://doi.org/10.1371/journal.pone.0252208
May 27, 2021 References Song QF, Liu XX, Hu WN, Han XC, Zhou WH, Lu AD, et al. Night Sleep Duration and Risk of Cognitive
Impairment in a Chinese Population: A Cross-sectional Study. Biomed Environ Sci. 2017; 30: 749–757. https://doi.org/10.3967/bes2017.100 PMID: 29122095 48. Bian Y. Bringing strong ties back in: Indirect ties, network bridges, and job searches in China. American
Sociological Review. 1997; 62: 366–385. https://doi.org/10.2307/2657311 49. Hang L, Zhang Y, Erpeng L, Nan X. How does social capital affect individual health among the elderly in
rural China?—Mediating effect analysis of physical exercise and positive attitude. PloS One. 2020; 15:
e0231318. https://doi.org/10.1371/journal.pone.0231318 PMID: 32716935 50. Yeh SC, Liu YY. Influence of social support on cognitive function in the elderly. BMC Health Serv Res. 2003; 3: 9. https://doi.org/10.1186/1472-6963-3-9 PMID: 12775218 51. Holtzman RE, Rebok GW, Saczynski JS, Kouzis AC, Wilcox Doyle K, Eaton WW. Social network char-
acteristics and cognition in middle-aged and older adults. J Gerontol B Psychol Sci Soc Sci. 2004; 59:
P278–284. https://doi.org/10.1093/geronb/59.6.p278 PMID: 15576855 52. Fratiglioni L, Wang HX, Ericsson K, Maytan M, Winblad B. Influence of social network on occurrence of
dementia: a community-based longitudinal study. Lancet. 2000; 355: 1315–1319. https://doi.org/10. 1016/S0140-6736(00)02113-9 PMID: 10776744 53. Berkman LF. Which influences cognitive function: living alone or being alone? Lancet. 2000; 355:
1291–1292. https://doi.org/10.1016/S0140-6736(00)02107-3 PMID: 10776738 54. Uchino BN, Holt-Lunstad J, Uno D, Betancourt R, Garvey TS. Social support and age-related differ-
ences in cardiovascular function: An examination of potential mediators. Ann Behav Med. 1999; 21:
135–142. https://doi.org/10.1007/BF02908294 PMID: 10499134 55. Zhang J, Lu N, Wang W. Does Education Moderate the Relationship between Social Capital and Cogni-
tive Function among Older Adults? Evidence from Suzhou City, China. Int J Environ Res Public Health. 2020; 17:6560. https://doi.org/10.3390/ijerph17186560 PMID: 32916917 56. Lockley S W, Skene D J, Arendt J. Comparison between subjective and actigraphic measurement of
sleep and sleep rhythms. Journal of Sleep Research. 1999; 8:175–183. https://doi.org/10.1046/j.1365-
2869.1999.00155.x PMID: 10476003 13 / 13
|
https://openalex.org/W3194585006
|
https://bovine-ojs-tamu.tdl.org/AABP/article/download/6877/6321
|
English
| null |
Surgery for the Urine Pooling Cow
|
American Association of Bovine Practitioners Conference
| 1,989
|
cc-by
| 1,389
|
0
"'O
(D
~
~
(")
(D
00
00
0..
......
00
,-+-
'"i
~
~
......
0 p Typically a urine pooler is an older cow which might
be expected to have a pendulous uterus. This type of indi-
vidual often has a tipped pelvis and may have somewhat of
a horizontal vulvar conformation. In our study most of the
cows were or had been embryo donors.1 This could indi-
cate a correlation with hormonal therapy but more likely
reflects the value of the cow and the continued effort to
make a diagnosis and treat this valuable cow. A urine pool-
er typically has a history of infertility characterized by fre-
quent breeding and no confirmed pregnancies. The vulvar lips must be retracted with stay sutures,
held by an assistant, or by use of a speculum such as a Cas-
lick's mare speculum. The vaginal mucosa is incised in an
elongated horizontal "U" along the floor of the vagina with
the apex of the incision located about 1 cm anterior to the
urethral orifice (Fig. 1 ). The sides of the incision extend
caudally on either side of the urethral orifice to approxi-
mately 2 cm from the vulvar lips. It is important that the
two sides of the incision are each about 2 cm from the ven-
tral commissure of the vagina. If these incisions are closer
to the floor of the vagina the resultant urinary tube will be
too small and may cause excessive straining in the post op-
erative period. The submucosa along the incision is under-
mined by sharp dissection to produce 2 mucosal layers. Examination reveals a pendulous uterus which is
often pulled over the brim of the pelvis on rectal examina-
tion. Also on rectal examination, urine ( usually in quanti-
ties of 1-2 quarts) can be manually expressed from the
anterior vagina with rectal message. There may or may not
be urine present in the uterus at the time of the examina-
tion. A vaginal exam reveals a varying quantity of urine
pooled in the anterior vagina, a more anterior position of
the urethral orifice than would be considered normal and
often a vaginitis secondary to the urine pooling. On vaginal
examination it is important to check for other causes of
infertility such as a prolapsed cervical ring or a cervical
tear. A uterine culture and / or an endometrial biopsy may
be indicated as part of the initial evaluation. 0
"'O
(D
~
~
(")
(D
00
00
0..
......
00
,-+-
'"i
~
~
......
0 p y
p
p
y
The ventral edges of the right and left incisions are
then apposed with 2-0 absorbable suture material with a
swaged, taper needle in a continuous Lembert suture pa-
tern (Fig. 2). These sutures need to be placed close togeth-
er and pulled snug to create an intimate apposition of
tissue which will prevent urine leakage which is very detri-
mental to healing and usually leads to fistula formation. This layer of closure essentially creates the urethral exten-
sion. The right and left dorsal mucosa! layers are then ap-
posed with the same suture material in a simple continuous
or horizontal mattress pattern (Fig. 3). This creates the
new ventral vaginal wall. With this extension the urine will
now be discharges about 2 cm from the vulvar lips and will
not spill or gravitate back into the vagina. Surgery for the Urine Pooling Cow Bruce L. Hull, DVM MS,
Department of Food Animal Medicine
The Ohio State University
Columbus, OH 43210 Bruce L. Hull, DVM MS,
Department of Food Animal Medicine
The Ohio State University
Columbus, OH 43210 Bruce L. Hull, DVM MS,
Department of Food Animal Medicine
The Ohio State University
Columbus, OH 43210 bovine is similar to the technique described for the equine
by Brown and Colahan.3 The urethroplasty for caudal ex-
tension of the urethra is performed with the cow in the
standing position. Local anesthesia is achieved by sacro-
coccygeal epidural injection of 5-7 ml of 2 % lidocaine. It is
important to evacuate the fecal material from the rectum
and to pack the rectum with a large piece of roll cotton. These two steps help prevent contamination of the peri-
nea} area during surgery. The fluid within the vagina is
manually evacuated and the vagina is rinsed with a mild
solution of tamed iodine. Once epidural anesthesia is com-
plete, the tail is tied to the side of the animal. The total
perinea! area is clipped and prepped in a routine manner. Before considering surgery for the urine pooler one
must first understand why urine pooling is detrimental and
what constitutes a diagnosis of a urine pooler. Doing sur-
gery for a urine pooler is of no benefit if the cow has other
pathology which is causing her infertility unless these other
problems are treated concurrently or in place of urine
pooler surgery. Surgical Procedure There has been a surgical procedure described to cre-
ate a fold of tissue in the vaginal floor just anterior to the
uretheral orifice. 2
This procedure utilizes a horizontal
mattress suture to gather a fold of vaginal mucosa just an-
terior to the urethral orifice. This fold serves as a dam to
help prevent anterior gravitation of the urine within the
vagina. Although this procedure may benefit some cows, it
has not given satisfactory results in cows with a severe tip-
ping of the pelvis or a very anterior position of the urethral
orifice. It needs to be noted that the most anterior end of the
incision (just above the urethral orifice) is very critical in
both the dissection and suturing. This area is very thin, be-
tween the urethra and ventral vaginal floor, and is often so
far cranial in the vagina that dissection and suturing are The surgery which we prefer for urine pooling in the THE BOVINE PROCEEDINGS-No. 22 12 FIGURE 1 difficult. However, this is also the area that is most prone
to failure and if a fistula develops in this area the surgery
will not be successful. FIGURE 1
FIGURE2
FIGURE3 FIGURE2 FIGURE2 0
"'O
(D
~
~
(")
(D
00
00
0.. ...... 00
,-+-
'"i
~
~
...... 0 p Post surgically the cow's micturition needs to be ob-
served closely, although if a tube of sufficient diameter has
been created this should be no problem. One is often
tempted to place an indwelling catheter post operatively,
however, the irritation caused by the catheter may cause
dehiscence or straining. The use of an indwelling catheter
is therefore not advisable. Post operatively, antibiotics are
probably indicated, although, the literature cites using an-
tibiotics or not using antibiotics as being up to individual
preference. As horizontal vulvar conformation may predispose a
cow to vaginal contamination with feces, the surgeon is
often tempted to perform a Caslick's in conjuction with the
urethral extension. Please note that a Caslick's procedure
in itself impedes the flow of urine and can be counterpro-
ductive to a urethral extension. Therefore, it is advisable to
wait and see if the urethral extension totally solves the
problem or if more surgery is needed. If a Caslick's must be
performed it should be a very minimal Caslick's and not
extend down over the newly created urethral orifice. Surgical Procedure Although it is necessary for correction of the urovagi-
na before successful breeding to take place, complete eval-
uation of the cow's reproductive tract should be performed
before surgery. The presence of chronic vaginitis, cervici-
tis, endometritis, cystic ovarian disease and other repro-
ductive problems must be assessed to allow a more
accurate prognosis for future breeding soundness. Surgical
failure of the urethral extension is often a result of these
secondary complications. Therefore, if they are not fully
evaluated and treated concurrently, a successful outcome
cannot be expected. FIGURE3 FIGURE3 FIGURE3 Bibliography 1. Saint-Jean G, Hull BL, Robertson JT, Hoffsis GF and
Haibel GK: Urethral Extension for the Correction of Ura-
vagina in the Bovine. A Review of 14 Cases. Veterinary Sur-
gery 17.5:258-262 1988. 2. Hudson RS: Repair of Perinea! Laceration in The Cow. The Bovine Practitioner. 5th Annual Convention. 34-40,
1972. 3. Brown MP, Callahan PT, and Hawkins, DL: Urethral
Extension for Treatment of Urine Pooling in Mares. Jour-
nal American Veterinary Medical Association. 173:1005-
1007, 1978. APRIL, 1990 13
|
https://openalex.org/W4316917506
|
https://jsr.lib.ums.ac.id/index.php/jkk/article/download/12/11
|
English
| null |
Women’s Right Violation in the Name of Islam Teaching A Feminist Essentialist Criticism Reflected on “A Thousand Splendid Suns” Novel
|
Jurnal Keilmuan dan Keislaman
| 2,022
|
cc-by
| 5,929
|
Keilmuan dan Keislaman
e-ISSN xxxx-xxxx Keilmuan dan Keislaman
e-ISSN xxxx-xxxx Abstract The topic of this study is women's human rights abuses in Afghanistan. Violations of
women's human rights were used as the main object of this study because the idea of
violating women's rights can be seen from the beginning to the end of the story of the novel
"A Thousand Splendid Suns" written by Khaled Hosseini. The purpose of this study is to
investigate more deeply the causes of unfair treatment for women in Afghanistan. This
research uses essentialist feminist theory to examine what are the types of violations of
women's rights and the causes of the unfair treatment received by women in Afghanistan. This theory will also be used to uncover what aspects cause society in Afghanistan to treat
women differently. The results of this study indicate that there are two types of violations
of women's human rights, namely domestic violence, and violations of social rights. The
study also indicates that Afghans’ misinterpretation of Islamic teachings is one of the
causes of women's rights abuses in Afghanistan. Keywords: Islamic teaching, Feminist Essentialist, Women’s rights abuses Keywords: Islamic teaching, Feminist Essentialist, Women’s rights abuses Article history: Submit: 5 Januari 2021; Revisi: 25 Januari 2022; Diterima: 10 Februari 2022
Publikasi: 1 Maret 2022; Periode Terbit: Maret 2022 Women’s Right Violation in the Name of Islam Teaching A Feminist Essentialist Criticism …. (42-51) Women’s Right Violation in the Name of Islam Teaching A
Feminist Essentialist Criticism Reflected on “A Thousand
Splendid Suns” Novel Women’s Right Violation in the Name of Islam Teaching A
Feminist Essentialist Criticism Reflected on “A Thousand
Splendid Suns” Novel Nur Indah Dwi Saputri1* 1Fakultas Keguruan dan Ilmu Pendidikan, Universitas Muhammadiyah Surakarta, Indonesia
*Corresponding email: A30140051@student.ums.ac.id akultas Keguruan dan Ilmu Pendidikan, Universitas Muhammadiyah Surakarta, Indonesia
*Corresponding email: A30140051@student.ums.ac.id Keywords: Islamic teaching, Feminist Essentialist, Women’s rights abuses Keilmuan dan Keislaman
e-ISSN xxxx-xxxx because it creates a subordination to
others. of all injustice, one of which is the issue
of discrimination against women. In fact,
the problem is a community tradition
that is not in accordance with Islamic
teachings, so it is more appropriate if the
problem of gender discrimination is
caused
by
religious
and
cultural
interpretation
factors
(Qomariah
&
Saputra, 2020). The issues of women’s inferiority in
this research are the reason why the
novel of A Thousand Splendid Suns by
Khaled Hosseini is selected to be the
research object of study because it
possesses a clear case of women’s
inequality treatment in society, whether
in the public places or private around
their family. A Thousand Splendid Suns
is a novel that tell a story of two women,
Mariam and Laila. They are destined to
meet each other by the hand of their
abusive husband, Rasheed, without
power to fight back. They live together to
bear Rasheed iron hand each day until
the point when Mariam cannot take it
anymore. Sacrificing her own life to give
Laila freedom is Mariam choice, because
of the law that strongly protect men no
matter what they do, Mariam statement
will never be counted in the court
resulting death penalty for her. The
reasons why the book A Thousand
Splendid Suns is chosen to be the object
of study of this research is the way
Khaled Hosseini portrays how women is
being treated on an Islam society. Khaled
Hosseini also success to bring his story to
be hearth breaking when he clearly
portrays how Laila and Mariam struggle
of
abuse
in
their
household
life,
especially Mariam. Apart from those two
reasons, this novel also depict interesting
issue as how brave Laila and Mariam
decide to rebel against the law and
traditions by trying to escape their hard
life. p
,
)
When a society made a label of how
women
and
men
should
behave
according to their gender, this condition
will create a ‘stereotypes’. Stereotypes
often made up by binary opposition
paradigm, which means a point of view
that separate the words into two
classifications
that
structurally
contradictory (Rohmaniyah, 2014). For
example, while women expected to be
home to rise their children, men should
work to provide their family, because
women are weaker, they should work
less than men. Women’s Right Violation in the Name of Islam Teaching A Feminist Essentialist Criticism …. (42-51) Introduction psychologically, and symbolically, are
widely disclosed and published through
the
media,
showing
the
low
understanding of adolescents on gender
awareness and justice (Sokowati, 2021). Gender inequality in fulfilling access to
basic needs can have a negative impact in
all sectors. This gap hinders the highly
qualified talents that exist in women
(Aktaria & Handoko, 2012). Gender
equality will decrease poverty, increase
the education levels that can bring the
world into more sustainable economy
(Firmansyah & Sihaloho, 2021). Religious
teachings are often regarded as the root Gender belongs to a social category
that refers to an individual's social,
cultural, and biological meaning and
identity (Astalini et al., 2021). Because
men
and
women
have
different
fundamental qualities, there is a choice
effect between the two (Marzal et al.,
2022). Gender is as societal expectations
about men and women that have been
constructed Through news, fiction or
advertising. In this case, Mass media
plays a role in gender socialization
(Dwita, 2018). Cases of sexual violence,
sexual
harassment
both
physically, 42 Keilmuan dan Keislaman
e-ISSN xxxx-xxxx Keilmuan dan Keislaman
e-ISSN xxxx-xxxx The assumption that the
main task of women is to serve their
husbands, their duties and functions are
only to carry out work related to house
work (Rahmayati et al., 2021). Also, there
are different social expectations for
women and men. From an early age,
women are associated with being non-
aggressive, subtle, dependent, passive,
and non-decision-making, whereas boys
are associated with being aggressive,
active, independent, decision-making,
and dominant (Subiyantoro et al., 2019). The characteristics of men stereotype is
showing
an
independent
character,
strong personality, and ambitious. The
stereotype for women includes cheerful,
spoiled, and obedient (Cunandar, 2019). The perception of men is higher than the
average value of women perception
because men tend to have critical
thinking
skills
towards
material
(Kurniawan et al., 2022). This kind of
stereotypes can be seen as discrimination This present study uses feminist
essentialist
criticism
as
underlying
theory. Simone de Beauvoir is the
proponent of Feminist Essentialist. De
Beauvoir believes if Otherness is the basic
category in human mind. She argues that
women identity as Other and their
isolation are based, partly, on her 43 Keilmuan dan Keislaman
e-ISSN xxxx-xxxx Muhammad Imran Joiya, et. al. and
Nurul Istikhomah. Both of the research
describes women attitude when they
respond
to
the
patriarchal
society
oppression. The last study is written by
D. Alice Ligoria. She explains that
religion is the main factor of women
inequal treatment in society. physique-especially their capability on
reproduction – and partly on division of
labor based on their sex determined by
their ability to conceive and raise a child
(Humm, 1990). According to Simone de
Beauvoir book, The Second Sex (1949) it
can be seen that women life is divided
into several stages. The first stage is
childhood where the society is the one
who put the label male or female to their
children. Marriage is the next stage of
women life where they have no equal
status with their husband. Their inequal
status then being emphasized by how the
society does not see the wife as the part
of the society as an individual. These
women are expected to serve society
within their house walls when they will
interact with each other without knowing
what actually happen outside. Not only
tormented by the restriction in the
society, they also have their own fear. These women fear to lose their husband
favor when age catch them. They fear to
lose everything when their husband does
not see them as beautiful as when they
are young. Nonetheless, as time pass,
society gives more understanding for
women. They finally allowed to feel a
little freedom. Method While conducting this research, the
researcher used qualitative research as
research
design
because
the
main
concern of this research is explaining the
cause
of
Afghan
people
inequal
treatment to women. The data and the
source of data in this research are written
document. The data taken in the novel,
are related to the issues of religion aspect
that influence Afghan society point of
view toward women and laws that
violated women’s right. The instrument
on this research is the data collected from
the novel. Since the data resource of this
research is a novel in other word a
document, the researcher used document
analysis as the method of collecting data. In this research the researcher used
content analysis to analyses the data. Content analysis is a research tool used to
determine the presence of certain words
or concepts within texts or sets of texts
(Busch, et al., 1994 – 2012, p. 2). Using several previous studies, this
research
aims
to
make
deeper
understanding on how women are being
treated unequally by the society. The first
previous study is from Dr. Nongmaithem
that talks about how strong women are in
their life of oppression, they will reach
the point that they will take their own
fate. Second study is written by Shengai
Li and Tingting Liu, with the purpose of
study is to raise people awareness on
how women in Afghanistan still treated
unequally. The next study comes from
Marzieh
Gordan
and
Areej
Saaf
Almutairy, they talk about women
resistance under patriarchal society. The
next study used as reference is come from Women’s Right Violation in the Name of Islam Teaching A Feminist Essentialist Criticism …. (42-51) Keilmuan dan Keislaman
e-ISSN xxxx-xxxx The reason why these
children cannot refuse the marriage is
their father or head of the family has the
final say in this decision and they can
only agree. The impact of early marriage
causes the quality of the household to not
be in a superior performance both in
terms
of
reproductive
health,
psychological and economic readiness of
the family, thus bringing the impact of
being
vulnerable
to
divorce,
and
neglecting the quality of their children's
education. Psychological maturity is
lacking, problem solving methods are
less thoughtful, doing homework is not
optimal. Emotions are not stable in
solving household problems that take
turns (Julijanto, 2015). Another reason is
some of Hadis in Islam teaching such as,
Hadis narrated by Abu Daud no. 2083,
Tirmidzi no. 1102, Ibnu Majah no. 1879
dan Ahmad 6: 66 “From ‘Aisyah, she
said, Rasulullah shallallahu ‘alaihi wa
sallam said, ‘A married women without
her guardian's permission then her
marriage is invalid, invalid, invalid. And
if they disputed then the government is a
guardian for women who do not have a
guardian’”. (Menikah Tanpa Izin dengan
Orangtua dalam Islam, 2018). This Hadis
explains that a woman who marries
without their guardian’s permission,
their marriage is invalid. Consequently, by Laila and Aziza. Their parents neglect
their obligation to gives their children
supervision, or protection in health,
security and morality (Family and
Children’s Right, 2008). The reason why
the parents failed to give their children
supervision is first, Laila’s mother forgets
to picking Laila so Laila is bullied by her
friends. The second reason is when
Aziza’s father, Rasheed, he gives very
different treatment to his children. When
he showers his son with present, he
wants to make his daughter to be a
beggar. He
also
sends
Aziza
to
orphanage and keep Zalmai, his son,
with the family when they have economy
crisis. The cause that responsible of this
discrimination is Quran 4:11 that talks
about a son is valued as two daughters in
receiving
inheritance. Maybe
these
people are interpreting the verse as son
should be valued more than daughter so
they must give more than daughter. This
interpretation cannot be right because
according to Toward Understanding the
Quran, the explanation of the Quran 4:11,
sons receive more than daughter because
they have responsibility to their family. They must feed their family, as women,
they can life from their husband income. Results and Discussions Based on the underlying theory
used, feminist essentialist, the finding
and discussion will be divided according
to women life stages. On each stage of
women life, the current researcher
discusses what kind of women right
violation and what is the cause of the
violation. The first stage of women life is
childhood. In their childhood they are
being
discriminated
by
treated
differently from boys. It is experienced 44 Women’s Right Violation in the Name of Islam Teaching A Feminist Essentialist Criticism …. (42-5 Keilmuan dan Keislaman
e-ISSN xxxx-xxxx right violation because they are forced to
marry in such a young age, or it can be
called as children marriage. Girls are
very
vulnerable
to
being
sexually
exploited not only because of the lack of
literacy in girls and the economic
conditions of the family, but also because
of their environment and social status. The lower the social status, especially the
family's social status, the higher the
potential to be manipulated, made into a
sexual object, because socially in a
powerless
condition
(Anggelia
&
Purwanti, 2020). The reason why these
children cannot refuse the marriage is
their father or head of the family has the
final say in this decision and they can
only agree. The impact of early marriage
causes the quality of the household to not
be in a superior performance both in
terms
of
reproductive
health,
psychological and economic readiness of
the family, thus bringing the impact of
being
vulnerable
to
divorce,
and
neglecting the quality of their children's
education. Psychological maturity is
lacking, problem solving methods are
less thoughtful, doing homework is not
optimal. Emotions are not stable in
solving household problems that take
turns (Julijanto, 2015). Another reason is
some of Hadis in Islam teaching such as,
Hadis narrated by Abu Daud no. 2083,
Tirmidzi no. 1102, Ibnu Majah no. 1879
dan Ahmad 6: 66 “From ‘Aisyah, she
said, Rasulullah shallallahu ‘alaihi wa
sallam said, ‘A married women without
her guardian's permission then her
marriage is invalid, invalid, invalid. And
if they disputed then the government is a
guardian for women who do not have a
guardian’”. (Menikah Tanpa Izin dengan
Orangtua dalam Islam, 2018). This Hadis
explains that a woman who marries
without their guardian’s permission,
their marriage is invalid. Consequently, right violation because they are forced to
marry in such a young age, or it can be
called as children marriage. Girls are
very
vulnerable
to
being
sexually
exploited not only because of the lack of
literacy in girls and the economic
conditions of the family, but also because
of their environment and social status. The lower the social status, especially the
family's social status, the higher the
potential to be manipulated, made into a
sexual object, because socially in a
powerless
condition
(Anggelia
&
Purwanti, 2020). Keilmuan dan Keislaman
e-ISSN xxxx-xxxx relationship with purpose to gain control
from others (Melinda Smith & Jeanne
Segal, 2018). The domestic abuse consists
of verbal, physical and sexual abuse. Verbal abuse is when Rasheed shaming
Mariam in front of Laila. He deliberately
tells Laila how Mariam is lower than
them because she is an illegitimate child
and come from a country side. The
second abuse is physical abuse, reflected
by how Mariam is afraid of Rasheed
body language, especially when he
touches his belt. Mariam alerted body
language shows how often Rasheed
abuse her, because her body reflects
automatically
when
Rasheed
show
aggression to hit Mariam. The last abuse
is sexual abuse. This abuse is depicted
when Laila forcefully accepts Rasheed
request of intercourse because she does
not want Rasheed to hit Mariam. Laila
wants to rescue Mariam from Rasheed
physical abuse because Rasheed accuses
Mariam to make Laila refuse his request
of intercourse. they do not have any right to refuse the
matchmaking from their family. g
y
Another forced marriage is caused
by
pregnant
out
of
matrimony. Premarital sexual behavior in adolescents
is behavior that is driven by sexual desire
with the opposite sex which is carried out
without
going
through
an
official
marriage process according to religion or
according to law (Yudhaprawira &
Uyun, 2018). This is happened to Laila
when she finds out that she is pregnant
and she accept Rasheed proposal to
avoid stoning to death punishment. The
base of this law is Hadis Sahih Muslim,
17: 4191 'Ubada b. as-Samit reported:
Allah's Messenger (may peace be upon
him) as saying: Receive (teaching) from
me, receive (teaching) from me. Allah has
ordained a way for those (women). When
an unmarried male commits adultery
with an unmarried female (they should
receive)
one
hundred
lashes
and
banishment for one year. And in case of
married male committing adultery with a
married female, they shall receive one
hundred lashes and be stoned to death
(Translation of Sahih Muslim, book 17:
The Book Pertaining to Punishments
Prescribed by Islam (Kitab Al-Hudud,
2011). This abuse may be caused by Quran
4:34 that talks about husband right to
punish their wife if their wife does
something
wrong. This
verse
interpretation may be used as the
justification of their abusive action. While
this verse gives the right for men to
punish
their
wife,
but
physical
punishment is not allowed. Keilmuan dan Keislaman
e-ISSN xxxx-xxxx That is the reason why men or boys
should receive the two portions of
women or girls’ inheritance. Second stage of women life is
marriage life. Women in Afghanistan
does not have teenage life stage because
they will be married off by their family if
the family see them ready to be married
off. Early marriage is a marriage that
happens too soon at a young age, which
is carried out by couples aged 18 years
and under, both male and female. This is
due to the low economic level, low
education and inadequate access to
information (Nurhayati & Kurniasasri,
2020). This case is included as women 45 (42-51) Women’s Right Violation in the Name of Islam Teaching A Feminist Essentialist Criticism …. (42-51) Keilmuan dan Keislaman
e-ISSN xxxx-xxxx It seems that the law for travelling
women is based on Fataawa al-Lajnah al-
Daa’imah (17/339): It is haram for a
woman to travel without a mahram in all
cases, whether the journey is long or
short (al-Munajjid, 2018). As quoted from
The Telegraph (2011) Professor Akhtar-
Ul-Wasay, former head of Islamic Studies
at Delhi's Jamia Milia Islamia University
said that the fatwa is no longer fit for
modern era. When the Hadis being
issued, the condition of the society is very
traditional. There
are
no
safe
transportation and there are still so many
wild animals so women cannot travel
alone
without
their
male
relatives
because they cannot protect themselves. Compared to the condition from the day
when the Hadis is being issued to the
Afghanistan
condition
around
1992
when the law is made the Hadis cannot
be applied anymore because there are
already has proper transportation for
women to travel alone. The law of
women travelling then not only violated
their right to freedom of movement, it is
also endangering them. When these
women travel because they want to
escape their abusive house, they will
being send back to be tortured more. The
chance of escape become none, because their interest in society and the way of
participation can be form of freedom of
association. Freedom of association can
be interpreted as every individual can
participate in groups in the society, either
formal
or
nonformal
(Freedom
of
Association, n.d.). The violation against
this right is when Rasheed forbids his
wives to interact with his friends and
forbid them to go out alone. Rasheed also
asks his wives to wear burqa when the
going out. This condition closes every
chance of his wives, Mariam and Laila, to
have a decent interaction with others. Once Laila meets her old friend, Tariq,
Rasheed punish her by hit her. The
reasons why Rasheed doing what he did
to his wives because he wants to protect
his pride as men and husband. He wants
to protect his wives from other men’s
stare as described by Quran 24:30 that
verse talks about how men should keep
their gaze toward women because the
gazing
at
women
freely
can
be
considered
as
adultery
(Towards
Understanding the Quran, n.d.). Keilmuan dan Keislaman
e-ISSN xxxx-xxxx According to
Toward Understanding the Quran, even
the Prophet Muhammad does not agree
on physical punishment and recommend
to hit with something that will not leave
any marks on body and not to hit across
the face. According to the novel story
line, what Rasheed do is wrong even
Prophet Muhammad, does not do it. )
As it can be seen both of the case of
forced marriage is a limited freedom of
choice. Freedom of choice or the right or
ability to choose whatever you want to
do or have (Freedom of choice, 2018). The
freedom of choice that violated is a
freedom to choose the one they want to
marry. As for Laila case, the law is
restricting the choice to have children
outside of wedlock and also threat people
life just because they are pregnant out of
wedlock. The next stage of women life, is
their social life. Women receive a lot of
right violation. The first right violation is
right to participate in society. It means
that every human being has right to voice In the stage of married life, women
also experience domestic abuse or a
violation against someone’s partner in
intimate relationship such as marital 46 Women’s Right Violation in the Name of Islam Teaching A Feminist Essentialist Criticism …. (42-51) Keilmuan dan Keislaman
e-ISSN xxxx-xxxx right to freedom of movement and
residence within the borders of each
State, 2) everyone has the right to leave
any country, including his own, and to
return to his country (Definitions of the
right to freedom of movement, 2009). The
case in the novel for violation against the
right to freedom of movement is when
Mariam and Laila try to escape to
Peshawar. It is very difficult for them to
try to escape because the current law of
Afghanistan stated that every woman
cannot leave their house alone without
any male relatives. When they are
caught, they are returned to their house. It seems that the law for travelling
women is based on Fataawa al-Lajnah al-
Daa’imah (17/339): It is haram for a
woman to travel without a mahram in all
cases, whether the journey is long or
short (al-Munajjid, 2018). As quoted from
The Telegraph (2011) Professor Akhtar-
Ul-Wasay, former head of Islamic Studies
at Delhi's Jamia Milia Islamia University
said that the fatwa is no longer fit for
modern era. When the Hadis being
issued, the condition of the society is very
traditional. There
are
no
safe
transportation and there are still so many
wild animals so women cannot travel
alone
without
their
male
relatives
because they cannot protect themselves. Compared to the condition from the day
when the Hadis is being issued to the
Afghanistan
condition
around
1992
when the law is made the Hadis cannot
be applied anymore because there are
already has proper transportation for
women to travel alone. The law of
women travelling then not only violated
their right to freedom of movement, it is
also endangering them. When these
women travel because they want to
escape their abusive house
they will right to freedom of movement and
residence within the borders of each
State, 2) everyone has the right to leave
any country, including his own, and to
return to his country (Definitions of the
right to freedom of movement, 2009). The
case in the novel for violation against the
right to freedom of movement is when
Mariam and Laila try to escape to
Peshawar. It is very difficult for them to
try to escape because the current law of
Afghanistan stated that every woman
cannot leave their house alone without
any male relatives. When they are
caught, they are returned to their house. Keilmuan dan Keislaman
e-ISSN xxxx-xxxx As the right to become witness it is
influenced by Holy Quran 2:282 “Two
witnesses from among your men. And if
there are not two men [available], then a
man and two women from those whom
you accept as witnesses-so that if one
woman errs, then the other can remind
her.” This interpretation affects how
women intelligent are seen in the public
eyes. Women are seen as less intelligent
than men because they are need two
females as replacement of one man
witness just like the quotes from Holy
Quran above. chance to escape. The law of women
travelling become worse when Taliban
regime come. The women who are
caught travelling alone will be beaten
before returned to their home. Another right violated in women
social life is right to public service. According to Cambridge Dictionary
public service is a service provided by the
government, such as hospitals, schools,
or the police. As seen in the data women
has limited access for public services. The
first public services that being limited for
women is police protection. The police
refuse to protect these women from the
abuse they have been through. The police
ignore women safety because the policy
stated that police will not interfere with
private family affairs. According to Dr. Taha Jaber Al-
Alwani the interpretation of the verse is
the actual women witness needed is just
one. Two women needed as witness
because if one woman become witness
the other will be the guarantor to remind
the actual witness if what she says is right
or not. This verse also happens in the era
when women are not the one who
manage the transaction as Holy Quran
2:282 is verse that discuss about financial
problem. The second woman witness
here is a guarantor to make sure that the
main witness said the truth without
reducing or adding any other statement
(Al-Alwani, n.d.). The second public service denied
for women is education. Education for
women is being denied because of the
perspective of the society see education
for women is not important. The next
public service is hospital. In the novel,
when Taliban take control over in
Afghanistan, the hospital for women is
being closed and only leaving one
hospital for women and they limit
resource for the hospital for women. Not
only ignoring the health for women,
Taliban regime also ignoring the proper
facility for women prison. Keilmuan dan Keislaman
e-ISSN xxxx-xxxx g
)
This also considered to be a
restriction of interaction because when
women go outside with their husband,
and meet another woman at the street
they will not recognize each other
because their body is covered. They also
cannot recognize each other husband
because they are being forbade to see
other men, even if these women’s
husband is friends. As described before,
women have no way of interaction. A
woman who are living in polygamy
family may have a better chance of
interaction because they can interact with
other wives, but women who live in
monogamous family, they can be seen as
prisoner because they have no way to
interact with others but their husband. Another right that being violated is
freedom of movement. Freedom of
movement here is 1) everyone has the 47 Keilmuan dan Keislaman
e-ISSN xxxx-xxxx The prison for
women
can
be
considered
as
uninhabitable, furthermore the prison
warden does not provide food for the
women prisoner. The last public service
is court. In Afghanistan women cannot
become witness for her own case if there
are no men witnesses. The judges see
women as less intelligent than men, so
women cannot be trusted when they
become the witness in the court. The situation described in the Holy
Quran verse cannot be applied in the
Mariam’s or other similar case. What if
the woman kills people in self-defense
when there are no witnesses. They will be
punished because the judge cannot trust
the sole women witness. They will be
sent to jail because they want to protect
themselves. In Mariam case, she is being
sentenced to death. Women’s Right Violation in the Name of Islam Teaching A Feminist Essentialist Criticism …. (42-51) Keilmuan dan Keislaman
e-ISSN xxxx-xxxx a Mahram, even if it is a Short Trip. Retrieved from Islam Question and
Answer:
https://islamqa.info/en/101520 a Mahram, even if it is a Short Trip. Retrieved from Islam Question and
Answer:
https://islamqa.info/en/101520 women right violation in the novel. Nonetheless, it cannot be forgotten about
the background of Afghanistan history
itself. The wrong interpretation of Islam
teaching that led women right violation
in the novel actually caused by the
society condition in that country. A
different belief of Islam teaching among
different tribal and their rejection toward
modern Islam teaching interpretation is
what makes women position in society is
lesser than men. Anggelia, A., & Purwanti, A. (2020). Kebijakan
Perlindungan
Anak
terhadap
Eksploitasi
Seksual
melalui Nikah Siri dalam Perspektif
Hukum Nasional di Indonesia. Jurnal Jurisprudence, 10(1), 109–126. Astalini, A., Darmaji, D., Kurniawan, D. A., & Wulandari, M. (2021). Male or
Female, Who is Better? Students’
Perceptions of Mathematics Physics
E-Module
Based
on
Gender. Indonesian Journal on Learning and
Advanced Education (IJOLAE), 3(3),
207–224. This novel also can be seen as
Khaled Hosseini’s way to tell the world
how helpless Afghan women life is. They
cannot seek help to the police because
they will not interfere with what men do
to their families. Women also cannot seek
help to others because they will be beaten
if they walk out from their house. They
also will be prisoned if they try to run
from their family, their main abuser. Furthermore, they also cannot defend
themselves in the court because they
cannot be witness for their own case. Beauvoir, S. d. (1949). Le Deuxième Sexe
(The Second Sex). Paris: Éditions
Gallimard. Cunandar,
D. (2019). Kemampuan
Mendengarkan dengan Modalitas
Auditori Berbasis Gender. Kajian
Linguistik Dan Sastra, 4(2), 138–143. g
( )
Definitions of the Right to Freedom of
Movement. (2009, September 1). Retrieved from Claiming Human
Right:
http://www.claiminghumanrights
.org/freedom_movement_definitio
n.html It can also be seen that Khaled
Hosseini
wants
to
brings
people
awareness about how some women still
struggle from discrimination. These
women being abused openly by the law
in their country and no way to escape. They need to be protected and freed from
the law and the mindset of the society
where they life. Dwita, D. (2018). Gender Equality in
Media
Television
(Semiotics
Analysis
of
Fair
and
Lovely
Advertisement Issue of Marriage or
Master Degree). Komuniti: Jurnal
Komunikasi dan Teknologi Informasi,
10(1), 37–43. Conclusion After the research on novel A
Thousand Splendid Suns by Khaled
Hosseini by using feminist essentialist, it
can be concluded that Islam teaching is
one of the strong factors that causes The cause of the imbalance right in
public services is also caused by Islam
teaching in Holy Quran 4:11, which
stated that woman is valued half of man. 48 Women’s Right Violation in the Name of Islam Teaching A Feminist Essentialist Criticism …. (42-51) Keilmuan dan Keislaman
e-ISSN xxxx-xxxx Ekonomi
Pembangunan:
Kajian
Masalah Ekonomi dan Pembangunan,
22(1), 12–21. Perception of e-Module Linear
Equations in Mathematics and
Physics. Indonesian
Journal
on
Learning and Advanced Education
(IJOLAE), 4(2), 92–106. ( )
Freedom of Association. (n.d.). Retrieved
from
Human
Right
House
Foundation:
https://humanrightshouse.org/we
-stand-for/freedom-of-
association/#Registrationofcivilsoc
iety Li, S., & Tingting, L. (2017). On the
Miserable Female's Fate from A
thousand Splendid Suns. Advances
in Social Science, Education and
Humanities, Volume 119, 4. Ligoria, D. A. (n.d.). Study of Marginalized
Afghan Women in Khaled Hosseini’s. Retrieved
from
ACADEMIA:
www.academia.edu Freedom of choice. (2018, May 18). Retrieved
from
Longman
Dictionary
of
Contemporary
English:
https://www.ldoceonline.com/dic
tionary/freedom-of- choice Freedom of choice. (2018, May 18). Retrieved
from
Longman
Dictionary
of
Contemporary
English:
https://www.ldoceonline.com/dic
tionary/freedom-of- choice Marzal, J., Chit, S. C., Elisa, E., Utomo, P. E. P., Kurniawan, D. A., & Sandra,
R. O. (2022). Lecturer Gender
Perspective with Online Thesis
Guidance Case Study Elista in
Jambi University. Indonesian Journal
on Learning and Advanced Education
(IJOLAE), 4(3), 191–208. Gordan, M & Almutairy, A. S. (2013). Resistance,
a
Facet
of
Post-
colonialism in Women Characters
of Khaled Hosseini's a Thousand
Splendid Suns. International Journal
of Applied Linguistics & English
Literature, Vol. 2 No.3, 8. Melinda Smith, M., & Jeanne Segal, P. (2018, March). Helpguide.org Trusted
Guide to Mental & Emotional Health. Retrieved from Domestic Violence
and Abuse Recognizing the Signs of
an
Abusive
Relationship
and
Getting
Help:
https://www.helpguide.org/articl
es/abuse/domestic-violence-and-
abuse.htm Hosseini, K. (2007). A Thousand Splendid
Suns. London: ATSS Publications. Humm, M. (1990). Dictionary of Feminist
Theory. Colombus:
Ohio
State
University Press. Istikomah, N. (2015). Women’s
Attitudes Towards Gender
Discrimination. Yogyakarta:
Universitas Negeri Yogyakarta. Humm, M. (1990). Dictionary of Feminist
Theory. Colombus:
Ohio
State
University Press. Istikomah, N. (2015). Women’s
Attitudes Towards Gender
Discrimination. Yogyakarta:
Universitas Negeri Yogyakarta. Menikah Tanpa Izin dengan Orangtua dalam
Islam. (2018, April 11). Retrieved
from Dalami Islam - Pusat Ilmu
Islam
Nusantara:
https://dalamislam.com/hukum-
islam/pernikahan/menikah-tanpa-
izin-dengan-orangtua-dalam-islam g
gy
Joyia, Muhammad Imran, e. a. (2017,
January 1). ISSN 1930-2940 Vol. 17. Retrieved from Language in India:
www.langageinindia.com Julijanto, M. (2015). Dampak Pernikahan
Dini dan Problematika Hukumnya. Jurnal Pendidikan Ilmu Sosial, 25(1),
62–72. g
g
Nelson, D. (2011, March 9). Muslim
Women 'should not Travel more than
48 Miles from Home without Male
Chaperone'. Retrieved from The
Telegraph:
https://www.telegraph.co.uk/ne
ws/worldnews/asia/india/837122
1/Muslim-
women-should-not- Public Service [Def. 1]. (n.d.). Cambridge
Dictionary. Retrieved
from
http://dictionary.cambridge.org/
dictionary/nglish/public-service Kurniawan, D. A., Astalini, A., Darmaji,
D., Tanti, T., & Maryani, S. (2022). References Aktaria, E., & Handoko, B. S. (2012). Ketimpangan
Gender
dalam
Pertumbuhan Ekonomi. Al-Alwani, D. T. (n.d.). The Testimony of
Women in Islamic Law. Retrieved
from
Al-
Hewar
Center:
http://www.alhewar.com/TahaTe
stimony.htm
l
d
(
l
) Aktaria, E., & Handoko, B. S. (2012). Ketimpangan
Gender
dalam
Pertumbuhan Ekonomi. Family and Children's Right. (2008). Retrieved
from
Humanium:
https://www.humanium.org/en/f
amily-and-childrensrights/ Al-Alwani, D. T. (n.d.). The Testimony of
Women in Islamic Law. Retrieved
from
Al-
Hewar
Center:
http://www.alhewar.com/TahaTe
stimony.htm Firmansyah, C. A., & Sihaloho, E. D. (2021). The Effects of Women
Empowerment
on
Indonesia’s
Regional Economic Growth. Jurnal y
al-Munajjid, S. M. (2018, April 11). A
Women should not Travel Except with 49 s Right Violation in the Name of Islam Teaching A Feminist Essentialist Criticism …. (42-51)
50 Keilmuan dan Keislaman
e-ISSN xxxx-xxxx Innovative
Learning:
Gender 50 Keilmuan dan Keislaman
e-ISSN xxxx-xxxx travel-more-than-48-miles-from-
home-without-male-
chaperone.html http://www.islamicstudies.info/t
afheem.php?sura=4&verse=13&to=
14 Translation of Sahih Muslim, book 17:
The Book Pertaining to Punishments
Prescribed by Islam (Kitab Al-Hudud). (2011). Retrieved from Center for
Muslim-Jewish
Engagement:
http://cmje.usc.edu/religious-
texts/hadith/muslim/017-
smt.php#017.4191 Nongmaithem, D. A. (2017). The Role of
Women in A Thousand Splendid Suns
by
Khaled. IOSR
Journal
of
Humanities and Social Science
(IOSR-JHSS), 4. Nurhayati, S. F., & Kurniasasri, I. (2020). Analisis
Pernikahan
Usia
Dini
Ditinjau
dari
Sudut
Pandang
Ekonomi, Sosial dan Religi: Studi
pada
Kecamatan
Purwodadi
Kabupaten
Grobogan. Profetika:
Jurnal Studi Islam, 21(1), 17–26. Yudhaprawira, M. R., & Uyun, Z. (2018). Kematangan
Beragama
Remaja
Akhir
sebagai
Pelaku
Seksual
Pranikah. Indigenous: Jurnal Ilmiah
Psikologi, 2(1). Qomariah, N. L., & Saputra, A. (2020). Penggunaan
dan
Penguasaan
Ruang Berbasis Gender di UMS. Sinektika: Jurnal Arsitektur, 17(2),
128–134. Quran English Android Application. (2018). Developed by Andiumpam.com Rahmayati, R., Ramadhan, S., & Afnita,
A. (2021). Diskriminasi Gender
dalam
Novel
Perempuan
Terpasung
Karya
Hani
Naqshabandi: Kajian Feminisme
Sastra. Kajian Linguistik dan Sastra,
6(1), 84–95. ( )
Rohmaniyah, I. (2014). Kontruksi
Patriarki dalam Tafsir Agama. Yogyakarta:
Universitas
Islam
Sultan Kalijaga Yogyakarta. Sokowati, M. E. (2021). Penguatan
Jurnalistik Berbasis Gender untuk
Kader Pimpinan Cabang Ikatan
Mahasiswa
Muhammadyah
AR
Fakhruddin. Warta LPM, 24(3), 436–
446. Subiyantoro, S., Wardani, N. E., &
Saddhono, K. (2019). Kedudukan
Wanita Jawa sebagai Istri dalam
Novel Alun Samudra Rasa Karya
Ardini
Pangastuti
BN. Kajian
Linguistik dan Sastra, 3(1), 77–86. Towards Understanding the Quran. (n.d.). Retrieved
from 51
|
https://openalex.org/W4230563361
|
http://www.scielo.br/pdf/bjb/v74n2/1519-6984-bjb-74-02-editorial.pdf
|
English
| null |
EDITORIAL NOTE
|
Brazilian Journal of Biology
| 2,014
|
cc-by
| 154
|
DOI: http://dx.doi.org/10.1590/1519-6984.740201 DOI: http://dx.doi.org/10.1590/1519-6984.740201 ISSN 1519-6984 Br a z i li a n Jo ur n a l o f Bi o lo g y ® MAY, 2014 MAY, 2014 NUMBER 2 EDITORIAL NOTE The Brazilian Journal of Biology is preparing three special numbers for 2014 and early 2015. One refers to the Belo Monte Reservoir located in the Xingu River in collaboration with Norte Energia. Two other special numbers are a collection of papers covering the Rio do Sinos watershed, elaborated
in collaboration with University Feevale. The distribution of printed volumes of the Brazilian Journal of Biology will be restricted to
Universities which have the annual subscription. Since the papers are easily accessed from SciELO, the
Editorial Board considers it sufficient for associates to access the whole content of the journal from that
site. Authors will continue to receive a printed copy of the journal that contains their published papers. The Editorial Board
|
https://openalex.org/W2969849271
|
https://www.int-arch-photogramm-remote-sens-spatial-inf-sci.net/XLII-4-W14/143/2019/isprs-archives-XLII-4-W14-143-2019.pdf
|
English
| null |
SMODERP2D SOIL EROSION MODEL ENTERING AN OPEN SOURCE ERA WITH GPU-BASED PARALLELIZATION
|
The international archives of the photogrammetry, remote sensing and spatial information sciences/International archives of the photogrammetry, remote sensing and spatial information sciences
| 2,019
|
cc-by
| 6,289
|
ABSTRACT: SMODERP2D is a runoff-soil erosion physically-based distributed episodic model used for calculation and prediction processes at
agricultural areas and small watersheds. The core of the model is a raster based cell-by-cell mass balance calculation which includes
the key hydrological processes, such as effective precipitation, surface runoff and stream network routing. Effective precipitation,
the forcing of the runoff and erosion processes, is reduced by surface retention and infiltration. Surface runoff consists of two
components: slower sheet and concentrated rapid rill flow. Stream network routing is performed line-by-line in the user predefined
polyline layer. SMODERP is a long-term project driven by the Department of Landscape Water Conservation at the Czech Technical University
in Prague. At the beginning, SMODERP has been developed as a surface runoff simulated by profile model (1D). Later the model
has been redesigned using a spatially distributed method. This version is named SMODERP2D. Ongoing development is focused
on obtaining parameters of the hydrological models, incorporating new infiltration and flow routing routines, and conceptualization
of a rill flow and rill development. The model belongs to a family of so-called GIS-based hydrological models utilizing capabil-
ities of GIS software for geospatial data processing. Importantly, the SMODERP2D project is currently entering an open source
world. Originally the model could be run only in proprietary Esri ArcGIS platform. A new version of the model presented by this
manuscript adds support for two key open source GIS platforms, GRASS GIS and QGIS. A newly developed GRASS module and
QGIS plugin significantly increases the accessibility of the SMODERP2D model for research purposes and also for engineering
practice. Middle scale distributed hydrological models often encounter with high computation costs and long model runtime. Long runtime
is caused by high-resolution input data which is easily available nowadays. The project also includes an experimental version of
the SMODERP2D model enabling the parallelization of computations. This parallelization is done using TensorFlow, and its goal
is to decrease the time needed for its run. It is supported by both CPU and GPU. Parallelization of computations is an important
step towards providing SMODERP2D web processing services in order to allow quick and easy integration to highly specialized
platforms such as Atlas Ltd. Commission IV, WG IV/4 Commission IV, WG IV/4 KEY WORDS: Hydrology, Soil Erosion Models, GIS, Open Source, Parallel computing This contribution has been peer-reviewed.
https://doi.org/10.5194/isprs-archives-XLII-4-W14-143-2019 | © Authors 2019. CC BY 4.0 License. SMODERP2D SOIL EROSION MODEL ENTERING AN OPEN SOURCE ERA WITH
GPU-BASED PARALLELIZATION M. Landa1, J. Jeˇr´abek2, O. Peˇsek1, P. Kavka2 1 Dept. of Geomatics, Faculty of Civil Engineering, Czech Technical University in Prague,
Czech Republic - (martin.landa, ondrej.pesek)@fsv.cvut.cz
2 Dept. of Landscape Water Conservation, Faculty of Civil Engineering, Czech Technical University in Prague,
Czech Republic - (jakub.jerabek, petr.kavka)@fsv.cvut.cz 1 Dept. of Geomatics, Faculty of Civil Engineering, Czech Technical University in Prague,
Czech Republic - (martin.landa, ondrej.pesek)@fsv.cvut.cz
2 Dept. of Landscape Water Conservation, Faculty of Civil Engineering, Czech Technical University in Prague,
Czech Republic - (jakub.jerabek, petr.kavka)@fsv.cvut.cz 2.1
SMODERP2D model outflow from a given cell [LT −1]
inf = infiltration [LT −1]
ret = surface retention for a given raster cell [LT −1] ret = surface retention for a given raster cell [LT −1] In the case of EH models, the commonly computed processes
are sheet and rill flow. The sheet flow covers the earth’s surface
evenly, whereas rill flow detaches the soil material and con-
centrates its flow in the created rill (therefore it is also called
concentrated flow). Although the concentrated rill flow is par-
ticularly fast (causing the time step size constraint) it usually
occupies a small portion of the area. The computation may end
up in a situation where a small portion of the computed area
demands a shorter time step (due to rill flow presence) whereas
the rest of the area allows larger time step. In that case, only
a small part of the computed area with developed rills causes
a long model run-time. The sum Pn
j in the expression (2) represents sum of all inflows
to the cell i. The flow direction and therefore the sum Pn
j is
controlled by D8 flow direction algorithm (O’Callaghan, Mark,
1984). Effective precipitation es is potential precipitation re-
duced by interception of the rainfall water on the vegetation. The model is forced to satisfy the Courant–Friedrichs–Lewy
(CFL) criterion (Courant et al., 1928): CFL = qdt
dx < 1.0
(4) (4) where
dt = time step [T]
dx = grid cell size [L] where where To summarize and outline the objectives of this manuscript. The
advantage of high-resolution geospatial data availability is con-
strained with an increasing computation demands of a calcula-
tion. In the case of this manuscript, the extremely short time
steps caused by the needs of CFL criterion is the main con-
cern. Not all computed processes need a shorter time step and
processes which are spatially limited (the concentrated flow in
rill). In other words, the whole basin computation run-time is
being increased due to a small part of the computed basin. One
way to overcome this problem is to use GPU or CPU-based
parallelization. In this manuscript, TensorFlow Python library
(Abadi et al., 2015) was tested to parallelize the EH model. Be-
sides the TensorFlow also a CPU-based parallelization is out-
lined. The testing was performed with the SMODERP2D EH
model. 2.
MATERIAL AND METHODS or points (typically a water pump) features may be presented in
the modelled system and affect the water flow or soil transport
regime. GIS software has tools to operate with the linear and
point features, and geospatial data which simplifies modellers
live. 1.
INTRODUCTION Linear (water courses, streets, ditches) Empirical erosion models are often based on Universal Soil
Loss Equation (USLE), (Wischmeier et al., 1978, Renard et al.,
1997) and empirical hydrological models on the Curve number
method (CN) (Cronshey, 1986), concepts more than 30 years 143 The International Archives of the Photogrammetry, Remote Sensing and Spatial Information Sciences, Volume XLII-4/W14, 2019
FOSS4G 2019 – Academic Track, 26–30 August 2019, Bucharest, Romania The International Archives of the Photogrammetry, Remote Sensing and Spatial Information Sciences, Volume XLII-4/W14, 2019
FOSS4G 2019 – Academic Track, 26–30 August 2019, Bucharest, Romania The International Archives of the Photogrammetry, Remote Sensing and Spatial Information Sciences, Volume XLII-4/W14, 2019
FOSS4G 2019 – Academic Track, 26–30 August 2019, Bucharest, Romania This contribution has been peer-reviewed.
https://doi.org/10.5194/isprs-archives-XLII-4-W14-143-2019 | © Authors 2019. CC BY 4.0 License.
144 This contribution has been peer-reviewed. 1.
INTRODUCTION old. Using empirical approaches may introduce limitations in
the protection measures design e.g. because mentioned models
do not take into account the transient nature of modelled pro-
cesses. Physically based models are being developed to over-
come the empirical models limitations. Erosion / hydrological models (EH) are being used for various
research or engineering purposes. Results of such models may
be used as input information for planning or designing soil con-
servation measures in the landscape and hydrological units -
basins. Runoff water volume and transported soil amounts or
discharge time series are being calculated in order to design the
protection measures sufficient for a given flood or soil transport
event. Another example of a practical application of EH mod-
els may be land-use change, build up areas development studies
or effect of those on water or soil transport regime. Great use
of EH models is also in extreme event forecasting. In research,
EH models are being used to proof a new theory or to test hy-
potheses related to mechanism controlling the runoff and soil
transport. Processes taking place in a landscape are spatially distributed,
which is the reason why GIS (Geographic Information System)
is often deploying in the modelling process taking advantage
of ready to use GIS features. EH models have similar struc-
ture (although each model is specific in the terms of processes
solved, its purpose or coding strategy). Runoff and soil loss are
initiated by precipitation which is, especially for larger areas,
spatially distributed process. Majority of models include an
infiltration routine with spatially distributed parameters, since
grassland and parking lot may be presented in a single hydro-
logical model and have vastly distinct infiltration characteris-
tic. Infiltrated water is transported to the soil which has varying
transport properties. Ponging water creates overland flow which
leads to a soil transport and may cause severe soil and nutrition
losses in the landscape. This contribution has been peer-reviewed. This contribution has been peer-reviewed. 2.1
SMODERP2D model The SMODERP2D model has been integrated in open source
GIS packages and tested for the GPU/CPU parallelization
within presented work. The model, which is now capable of
2D calculation, has been developed from the 1D profile ver-
sion (Hol´y, 1984). Description of the model follows. The EH model may encounter with some run-time issues which
rise from model spatial and temporal discretization. Data avail-
ability and larger computation resources lead more often to the
use of finer spatial resolution. It was noted in (Molnar, Julien,
2000) that raster grid cell size is interchangeable in the terms of
a spatial discretization if the model parameters were calibrated
on the model with the same raster grid size. Finer spatial resolu-
tion, in some cases, causes problems with a time discretization
and the time step size. Time step size is commonly controlled
with Courant–Friedrichs–Lewy (CFL) criterion (Courant et al.,
1928). CFL criterion forces the time step to decrease if: a) ve-
locity of flow process increases or; b) the spatial discretization
becomes finer. Maximum acceptable CFL value, which pre-
serves computation stability, theoretically equals one. For shal-
low surface processes (processes which take place in the used
model) CFL criterion should be even smaller than one as it was
noted in (Zhang, Cundy, 1989) or (Esteves et al., 2000). The
need for smaller than one CFL criterion is caused mainly by
the discrepancy between a solution (surface water height), cell
size and surface roughness coefficient or by sharp surface slope
changes between adjacent cells. The model has a simple structure based on the mass balance
equation: Storage
∆t
= Inflow −Outflow
(1) (1) where Storage represents surface water level h [L] which
changes each proceeding time during the computation. Inflow
and Outflow terms on the right-hand side of the equation (1)
represent the water flowing in and out the storage during the
time step ∆t and consist of several components. The Inflow
and Outflow of ith raster cell are defined as: Inflowi = esi +
n
X
j
qj
(2) Inflowi = esi +
n
X
j
qj
(2)
Outflowi = infi −qi −reti
(3) (2) Outflowi = infi −qi −reti
(3) (3) where
es = effective precipitation [LT −1] where
es = effective precipitation [LT −1] where
es = effective precipitation [LT
]
q = inflow to resp. 2.1
SMODERP2D model The model calculates the surface runoff and soil loss
processes with the use of GIS software for the data pre- and
postprocessing. GRASS GIS provider and QGIS plugin were
lately implemented in the SMODERP2D project, next to the
already existing Esri ArcGIS Toolbox. Those new features and
some of the principles used in the SMODERP2D model are also
presented in this manuscript. If the flow q is high, the model is forced to decrease the time
step in order to satisfy the CFL criterion, since a grid cell size
is fixed. The flow q in the equations (2) and (3) has two components. Slower and spatially extensive sheet flow qsh: qsh = XIY hb
(5)
X, Y, b = empirical parameters [−]
I = surface slope [−] (5) X, Y, b = empirical parameters [−]
I = surface slope [−] where and faster concentrated rill flow qrl calculated by the Mannings
formula: qrl = A 1
nR2/3I1/2
(6) (6) where
A = cross-section area [L2]
n = roughness in the rill [TL−1/3]
R = hydraulic radii [L] where
A = cross-section area [L2]
n = roughness in the rill [TL−1/3]
R = hydraulic radii [L] This contribution has been peer-reviewed. https://doi.org/10.5194/isprs-archives-XLII-4-W14-143-2019 | © Authors 2019. CC BY 4.0 License. 144 144 The International Archives of the Photogrammetry, Remote Sensing and Spatial Information Sciences, Volume XLII-4/W14, 2019
FOSS4G 2019 – Academic Track, 26–30 August 2019, Bucharest, Romania The International Archives of the Photogrammetry, Remote Sensing and Spatial Information Sciences, Volume XLII-4/W14, 2019
FOSS4G 2019 – Academic Track, 26–30 August 2019, Bucharest, Romania The International Archives of the Photogrammetry, Remote Sensing and Spatial Information Sciences, Volume XLII-4/W14, 2019
FOSS4G 2019 – Academic Track, 26–30 August 2019, Bucharest, Romania The resulting flow is a sum of sheet and rill flow: has been separated into new classes. This step was crucial in
order to make data preparation workflow GIS package indepen-
dent. The only supported platform, Esri ArcGIS, has been sep-
arated from the base workflow. Based on that, a new concept of
so-called GIS providers has been introduced, see Figure 2. q = qrl + qrl
(7) (7) The sheet flow starts when the infiltration capacity is exceeded;
when rainfall is higher than infiltration. The rill flow emerges
if a critical water level of sheet flow is exceeded. 2.2
SMODERP2D entering an open source world SMODERP2D is the project with a long history. Over the years
its development has been driven by the Department of Land-
scape Water Conservation at the Czech Technical University in
Prague (see SMODERP2D logo in Figure 1). In 2018 SMOD-
ERP2D developers started working on a new generation of the
model in order to solve or at least to improve various critical
issues of the project. This includes most importantly the com-
putation stability and performance, better interoperability, and
lack of documentation. Recently SMODERP2D source code
has been published on GitHub (SMODERP2D Development
Team, 2019) under GNU GPL licence in order to attract a wider
audience, new developers and users. Crucial is the separation of GIS functionality related code from
the generic workflow defined by the base provider. The base
provider depends only on standard builtin Python libraries. Array-like computation is performed by a well-known NumPy
library. Using GIS provider prototypes, the SMODERP2D
project can be easily extended to support other GIS packages. Currently, the SMODERP2D project comes with three differ-
ent GIS providers. Support for Esri ArcGIS platform is im-
plemented by ArcGISProvider, GRASS GIS is handled by
GrassGisProvider, see 2.2.1 for details. The CmdProvider
is triggered only when the model computation is run from
a command-line. In this case, it is assumed that the data prepa-
ration phase has been already performed by one of the sup-
ported GIS platforms. @ @ @
@
@
@ @
@ @ @
@ @ @ @
@ @ @
@ @ @
@
@ @
@ @
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@ @
@
@
@
@
@
@
@ @ @
@ @ @
@ @ @
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@
@ @ @
@
@
@ @
@ @ @
@ @ @ @
@
@
@
\
\
/
/ /
\
\
/
\
/
/
/
@ @ @
@ @ @
\ _\/
/_/
\
\/
\/
/_____/
@
@
@
@
\__/
\
/
_\___/
@
@
@
\____
\/
/
@
@
@
\_____/______/
@
@
@
\
@
@
@
\____________________ @ @ @ @
@ @ @
Figure 1. 2.1
SMODERP2D model The critical
water level is defined based on critical shear stress; when the
drag force of the flowing water becomes large than the cohesive
forces of the soil particles. From the definition, the sheet flow
does not occur all over the basin area. The rill flow is usually
presented to even lower extend. However, the CFL criterion is
more likely constrained by the rapid rill flow even though its it
occupy smaller area compared to sheet flow. Figure 2. Concept of GIS providers (software dependencies
outlined by stereotypes) Infiltration is solved with Phillip’s infiltration equation (Philip,
1957): inf = 1/2St−1/2 + Ks
(8) (8) where Parameters of relations (5) (6) and (8) are in the most cases spa-
tially distributed. It is therefore beneficial to incorporate GIS
packages in the modeling process. Figure 2. Concept of GIS providers (software dependencies
outlined by stereotypes) 2.2
SMODERP2D entering an open source world ASCII-art SMODERP2D project logo Example of running computation from a command-line below. Option –typecomp roff specifies that only model computation
without data preparation phase is triggered. It means that data
has been already preprocessed and stored in a pickle file dis-
tributed by a test.ini configuration file. python ./bin/start-smoderp2d.py --typecomp roff \
--indata tests/test.ini python ./bin/start-smoderp2d.py --typecomp roff \
--indata tests/test.ini This contribution has been peer-reviewed.
https://doi.org/10.5194/isprs-archives-XLII-4-W14-143-2019 | © Authors 2019. CC BY 4.0 License. python ./bin/start-smoderp2d.py --typecomp roff \
--indata tests/test.ini Newly added option next edge al-
lows adding information about the next left and right edge based
on the segment orientation determined from surface slope. This
functionality is important for SMODERP2D in order to deter-
mine stream network correct connectivity as Figure 3 shows. 2.2.2
QGIS plugin
Recently the SMODERP2D model has
been integrated also into QGIS environment. QGIS1 is a widely
used open source GIS platform which can be easily extended
by user-defined plugins. A SMODERP2D QGIS plugin al-
lows performing both data preparation and model computation
phases in QGIS native environment, see Figure 5. Data pre-
processing is ensured by GRASS GIS provider as described
in 2.2.1. Note that QGIS installation normally comes with
GRASS GIS included. It means that GRASS dependency is
solved by QGIS installation itself. Experimental code of the
plugin compatible with the current long term release QGIS
version 3.4 is available from the project GitHub repository
(SMODERP2D Development Team, 2019). GNU GPL licence. These improvements have been integrated
into main distribution and will be part of upcoming GRASS
GIS version 7.8. A GRASS v.to.points module (GRASS Devel-
opment Team, 2019b) has been extended to extract from lines
start or end nodes only. This functionality is used to determine
the slope of a polyline stream feature to ensure that its direc-
tion will always be downslope. Another improvement is re-
lated to a v.to.db GRASS module (GRASS Development Team,
2019a). This tool allows uploading geometry-related informa-
tion into the attribute table. Newly added option next edge al-
lows adding information about the next left and right edge based
on the segment orientation determined from surface slope. This
functionality is important for SMODERP2D in order to deter-
mine stream network correct connectivity as Figure 3 shows. Figure 5. SMODERP2D model implemented as QGIS plugin Figure 3. Stream segmentation procedure network connectivity Figure 3. Stream segmentation procedure network connectivity Figure 5. SMODERP2D model implemented as QGIS plugin On the top of the GRASS GIS provider a specialized GRASS
r.smoderp2d module has been designed. This tool allows the
user running SMODERP2D model computation directly from
GRASS GIS working environment as demonstrated in Figure 4. The module can be easily installed in GRASS GIS similarly
to other extensions (so-called addons modules) by g.extension
command. By default, the r.smoderp2d module performs data
preparation phase followed by model computation steps. Data
preparation only can be performed by -d flag. python ./bin/start-smoderp2d.py --typecomp roff \
--indata tests/test.ini Figure 1. ASCII-art SMODERP2D project logo 2.2.1
GRASS GIS integration
SMODERP2D supported
GIS platforms have been recently extended by a new GRASS-
based GIS provider. Introducing an open source GIS platform
to SMODERP2D workflow is crucial from the perspective of
interoperability. SMODERP2D users can choose between a
proprietary Esri ArcGIS platform and an open source GRASS
GIS (Neteler et al., 2012). The GRASS GIS provider is de-
signed similarly to ArcGIS provider. From a Python perspec-
tive, there is only one difference, GIS functions are accessed by
PyGRASS package (Zambelli et al., 2013). Nevertheless, an in-
tegration of GRASS tools in the SMODERP2D project required
a few improvements in GRASS GIS itself. That was possible
since GRASS GIS is an open source project distributed under The model is implemented in Python programming language
using the object-oriented paradigm. The original source code
has been designed with a low level of scalability, limited read-
ability and interoperability. Part of the computation phase re-
sponsible for a data preprocessing was restricted to the sin-
gle platform only, Esri ArcGIS. In 2018 the original source
code has been completely refactorized. Python classes defining
computational steps were re-organized in a hierarchical manner. Major design-related changes have been done in Python classes
responsible for data handling and preparation using GIS soft-
ware tools. Data preparation workflow is handled by a newly-
defined a base, partly abstract Python class (BaseProvider in
Figure 2). Functionality depending on the used GIS package 145 The International Archives of the Photogrammetry, Remote Sensing and Spatial Information Sciences, Volume XLII-4/W14, 2019
FOSS4G 2019 – Academic Track, 26–30 August 2019, Bucharest, Romania The International Archives of the Photogrammetry, Remote Sensing and Spatial Information Sciences, Volume XLII-4/W14, 2019
FOSS4G 2019 – Academic Track, 26–30 August 2019, Bucharest, Romania GNU GPL licence. These improvements have been integrated
into main distribution and will be part of upcoming GRASS
GIS version 7.8. A GRASS v.to.points module (GRASS Devel-
opment Team, 2019b) has been extended to extract from lines
start or end nodes only. This functionality is used to determine
the slope of a polyline stream feature to ensure that its direc-
tion will always be downslope. Another improvement is re-
lated to a v.to.db GRASS module (GRASS Development Team,
2019a). This tool allows uploading geometry-related informa-
tion into the attribute table. 1https://www.qgis.org
2https://legacy.python.org/dev/peps/pep-0373/ This contribution has been peer-reviewed.
https://doi.org/10.5194/isprs-archives-XLII-4-W14-143-2019 | © Authors 2019. CC BY 4.0 License. 2.3
Parallel computing experiments In (Vivoni
et al., 2011) the basin was separated in sub-basins based on
stream network. The sub-basins communicated with each other
through so-called ghost cell. The strategy aimed to generate as
few ghost cells as possible; to reduce the communication be-
tween the CPU cores. This experimental SMODERP2D branch is still under develop-
ment; however, the alpha-version is ready to be used. Table 1
presents the results of different tests made on this version (com-
paring parallelized GPU computation, parallelized CPU com-
putation and a single CPU one). As can be seen in the table, the usage of GPUs is not always
the right way even when compared with CPUs, both single and
parallelized ones. The bottleneck of TensorFlow is its graph
initialization; this step is very time-consuming and therefore
can last many times longer than the computation itself for ex-
tremely small data. Another bottleneck is the memory shift be-
tween RAM and GPU virtual memory which concludes into
slower processes for weaker GPUs (compared with parallelized
computations being run on CPUs). Generally, for data of com-
mon size was the process with parallelized computations faster
(reaching around 60 per cent of the total computation time on
different architectures). Interesting moment is slower run of
much stronger CPU4 when compared with weaker CPU2; this
behaviour has to be examined deeper. The parallelization strategy outlined in the manuscript is based
on the hydrological reality and it is shown in a simplified setup
in Figure 7. In this example, the Nuˇcice experimental catch-
ment was chosen to present the parallelization strategy. At
this 0.5 km2 large basin a long-term monitoring of erosion and
runoff processes is being conducted by the Dept. of Landscape
Water Conservation. The strategy main goal is the reduction of the communication
between CPU-cores during the computation as much as possi-
ble. The whole basin is divided into several sub-basins based
on the digital elevation model and user-defined sub-basin size. Outlet4 of each sub-basin is depicted with red dots in Figure 7. After the sub-basins are defined, an order in which each sub-
basin will be computed is defined as follows. Sub-basins which
are hydrologically the farthest from the basin outlet (depicted
by the triangle in Figure 7) and therefore have no inflow flow
upslope area are calculated at first. Those sub-basins have the
rainfall stored in hyetographs as the only input. 3https://ai.google/research/teams/brain/ 2.3
Parallel computing experiments Because one of the most crucial points of SMODERP2D com-
putations is the speed, an experimental branch allowing (both
CPU and GPU-based) parallelized computations has been de-
veloped. The main step was to rewrite all loop-based computations into
matrix-based mathematical operations. To keep matrices as so-
called tensors and to perform all the operations, an open source
TensorFlow Python library (Abadi et al., 2015) developed by
Google Brain Team3 was used. Even though TensorFlow is
most widely used for machine learning and its performance on
basic mathematical operations is not always better than the one
of NumPy (a quick comparison with NumPy and Numba can be
seen in (Puget, 2015)), it had been preferred for its easy switch
between CPU and GPU-based core (it depends only on the ver-
sion of TensorFlow the user has installed, no needs for changes
in the code) and therefore support also for users without an ac-
cess to machines with GPU. Another advantage of TensorFlow
is its usage of so-called graphs. A graph is a representation of
all operations in dataflow/workflow. Its individual operations
are automatically sent to multiple cores in a CPU or multiple
threads in a GPU. These nodes are run independently in paral-
lel. Table 2. Used processing units
ID
Model
Clock speed
Memory
GPU1
GeForce GTX 1060
3GB
33 MHz
3,016 MiB
GPU2
4× GeForce GTX
1080 Ti
33 MHz
11,178 MiB
CPU1
AMD
Ryzen
7
1700
Eight
Core
Processor
1.373 GHz
512 KB
CPU2
16× AMD Ryzen
7 1700 Eight Core
Processor
1.373 GHz
512 KB
CPU3
Intel
Xeon
CPU
E5-2630 v4
2.4 GHz
25,600 KB
CPU4
40×
Intel
Xeon
CPU E5-2630 v4
2.4 GHz
25,600 KB Table 2. Used processing units To support further development of TensorFlow and exploit its
bleeding edge functionalities, TensorFlow 2.0, which is pub-
lished currently just as an alpha version, was used in the
SMODERP2D experimental branch. Because TensorFlow 2.0
is still not suitable with all the Python acrobatic tricks, NumPy
was used for matrix operations in places where TensorFlow
could not (on places where loops were still needed; looping
through a NumPy array is incomparably faster than through
a Tensor). into sub-domains based on a certain algorithm where each sub-
domain computation is loaded to a single CPU core. It is benefi-
cial to incorporate the hydrological behaviour in the paralleliza-
tion strategy if the domain is a hydrological basin. 4The location in the basin where all water from the basin flows python ./bin/start-smoderp2d.py --typecomp roff \
--indata tests/test.ini In this case, the
module creates a binary pickle file which can be later used for
a subsequent model computation. Note that ArcGIS Toolbox
also allows creating a pickle file for later usage. Importantly,
such pickle files are platform independent. 2.2.3
Python3 support
SMODERP2D project also comes
with Python 3 support, but still supporting Python 2. Note
that Python versions 2 and 3 are not backwards compati-
ble. Python 3 support is important from various perspectives. Python 2 is slowly reaching the end of life2, but still used by
many GIS platforms such as Esri ArcGIS 10.x. Newly sup-
ported GIS platforms by the SMODERP2D project as Esri
ArcGIS Pro, (upcoming) GRASS GIS 7.8 and QGIS 3.x are
Python 3 based. On the other hand it is still meaningful to
support both Python versions, Python 2 mainly because of Esri
ArcGIS 10.x platform. Figure 4. Running r.smoderp2d module from GRASS GIS
graphical user interface Figure 6. SMODERP2D model available as ArcToolbox for Esri
ArcGIS 10.x and Pro platforms Figure 4. Running r.smoderp2d module from GRASS GIS
graphical user interface Figure 6. SMODERP2D model available as ArcToolbox for Esri
ArcGIS 10.x and Pro platforms r.smoderp2d command-line usage example: r.smoderp2d elevation=w001001 soil=soil_map \
soil_type=Novak vegetation=soil_map \
vegetation_type=veg rainfall_file=rainfall.txt \
points=points2 table_soil_vegetation=tab_sv \
table_soil_vegetation_code=soilveg \
table_stream_shape=tab_stream_shape \
table_stream_shape_code=smoderp stream=stream 146 The International Archives of the Photogrammetry, Remote Sensing and Spatial Information Sciences, Volume XLII-4/W14, 2019
FOSS4G 2019 – Academic Track, 26–30 August 2019, Bucharest, Romania The International Archives of the Photogrammetry, Remote Sensing and Spatial Information Sciences, Volume XLII-4/W14, 2019
FOSS4G 2019 – Academic Track, 26–30 August 2019, Bucharest, Romania Table 1. Results of parallelization tests
RAM
Processing unit
Data 62 KB
Data 197 MB
[s]
[s]
15 GB
GPU1
4.0
7,560
CPU1
0.2
12,809
CPU2
2.1
7,249
251 GB
GPU2
2.5
6,611
CPU3
0.2
10,637
CPU4
1.5
8,631 Table 1. Results of parallelization tests This contribution has been peer-reviewed.
https://doi.org/10.5194/isprs-archives-XLII-4-W14-143-2019 | © Authors 2019. CC BY 4.0 License. 2.3
Parallel computing experiments In the simpli-
fied setup shown in Figure 7, the sub-basins 1, 2, 3, and 6 are 2.3.1
Further ideas for a sub-basins based parallel com-
puting
Besides the GPU-based parallelization (with Ten-
sorFlow (Abadi et al., 2015) or NVIDIA Cuda technol-
ogy (Kalyanapu et al., 2011, Le et al., 2015)) the pure CPU
parallelization may also bring a good improvement in the com-
putation time reduction. The computation domain is separated 4The location in the basin where all water from the basin flows 147 The International Archives of the Photogrammetry, Remote Sensing and Spatial Information Sciences, Volume XLII-4/W14, 2019
FOSS4G 2019 – Academic Track, 26–30 August 2019, Bucharest, Romania The International Archives of the Photogrammetry, Remote Sensing and Spatial Information Sciences, Volume XLII-4/W14, 2019
FOSS4G 2019 – Academic Track, 26–30 August 2019, Bucharest, Romania The International Archives of the Photogrammetry, Remote Sensing and Spatial Information Sciences, Volume XLII-4/W14, 2019
FOSS4G 2019 – Academic Track, 26–30 August 2019, Bucharest, Romania The International Archives of the Photogrammetry, Remote Sensing and Spatial Information Sciences, Volume XLII-4/W14, 2019
FOSS4G 2019 – Academic Track, 26–30 August 2019, Bucharest, Romania Figure 7. Simple example of possible CPU-based parallelization
strategy for the experimental catchment Nuˇcice hoc solution should be chosen depending on the data and avail-
able computing power. Even though the SMODERP2D model
does not belong in the family of forecasting models (where the
short run-time is necessary) the run-time speed up will increase
the usability of the model in practice and research applications. hoc solution should be chosen depending on the data and avail-
able computing power. Even though the SMODERP2D model
does not belong in the family of forecasting models (where the
short run-time is necessary) the run-time speed up will increase
the usability of the model in practice and research applications. SMODERP2D source code is available on GitHub (SMOD-
ERP2D Development Team, 2019) under GNU GPL licence. SMODERP2D source code is available on GitHub (SMOD-
ERP2D Development Team, 2019) under GNU GPL licence. ACKNOWLEDGEMENTS The research has been supported by the research grants
TJ01000270,
QK1910029,
and
internal
CTU
grant
SGS17/173/OHK1/T3/11. Figure 7. Simple example of possible CPU-based parallelization
strategy for the experimental catchment Nuˇcice REFERENCES calculated at first in parallel. The calculated hydrographs of
the sub-basins 1, 2, 3, and 6 are stored in the memory for the
later use. Sub-basins which have sub-basins 1, 2, 3, and 6 in
its upslope area are calculated next. It this case it is the only
the sub-basin 4. The water inputs in the sub-basin 4 are now
hyetograph and also hydrographs of upslope sub-basins 1 and
3. Next sub-basin to be calculated is the final sub-basin 5. In
this sub-basin is the outlet of the whole area. The water input
in the final sub-basin 5 are hyetograph and hydrographs of sub-
basins 2, 4 and 6. Once the main outlet hydrograph is obtained
the calculation stores the results and stops. Abadi, M., Agarwal, A., Barham, P., Brevdo, E., Chen, Z.,
Citro, C., Corrado, G. S., Davis, A., Dean, J., Devin, M., Ghe-
mawat, S., Goodfellow, I., Harp, A., Irving, G., Isard, M., Jia,
Y., Jozefowicz, R., Kaiser, L., Kudlur, M., Levenberg, J., Man´e,
D., Monga, R., Moore, S., Murray, D., Olah, C., Schuster, M.,
Shlens, J., Steiner, B., Sutskever, I., Talwar, K., Tucker, P., Van-
houcke, V., Vasudevan, V., Vi´egas, F., Vinyals, O., Warden, P.,
Wattenberg, M., Wicke, M., Yu, Y., Zheng, X., 2015. Tensor-
Flow: Large-scale machine learning on heterogeneous systems. Software available from tensorflow.org. Courant, R., Friedrichs, K., Lewy, H., 1928. ¨Uber die partiellen
Differenzengleichungen der mathematischen Physik. Mathe-
matische annalen, 100(1), 32–74. This approach may encounter several limitations. The main one
originates from the basin geometry. In the case of a narrow
basin situation (each sub-basin has a single upslope and downs-
lope sub-basin), the sub-basins will be computed in a sequence,
which loses the advantage of multi-core working station. If this
situation happens the user will be forced to create very small
sub-basins in order to be able to perform the outlined CPU par-
allelization. The possibilities of CPU parallelization described
in this section will be the subject of further research. Cronshey, R., 1986. Urban hydrology for small watersheds. Technical report, US Dept. of Agriculture, Soil Conservation
Service, Engineering Division. Esteves, M., Faucher, X., Galle, S., Vauclin, M., 2000. Over-
land flow and infiltration modelling for small plots during un-
steady rain: numerical results versus observed values. Journal
of hydrology, 228(3-4), 265–282. This contribution has been peer-reviewed.
https://doi.org/10.5194/isprs-archives-XLII-4-W14-143-2019 | © Authors 2019. CC BY 4.0 License. This contribution has been peer-reviewed.
https://doi.org/10.5194/isprs-archives-XLII-4-W14-143-2019 | © Authors 2019. CC BY 4.0 License. 3.
CONCLUSION GRASS Development Team, 2019a. v.to.db GRASS mod-
ule. Geographic Resources Analysis Support System (GRASS)
Software, Version 7.7. Open Source Geospatial Foundation. https://grass.osgeo.org/grass77/manuals/v.to.db.html (2 June
2019). This manuscript presents SMODERP2D project and related re-
cently triggered development. SMODERP2D computational
tools have been successfully integrated into Esri ArcGIS,
GRASS GIS and QGIS desktop GIS platforms. On the top
of that, the concept of so-called GIS providers has been in-
troduced. Ongoing development is mainly focused on compu-
tational routines and parallel computation experiments. Also
OGC Web Processing Service providing SMODERP2D func-
tionality is planned to be established. All the tools are currently
distributed as experimental ready for testing and user feedback. The official stable release of SMODERP2D model is planned
for 2020. This includes also user documentation which is cur-
rently under development. GRASS Development Team, 2019b. v.to.points GRASS mod-
ule. Geographic Resources Analysis Support System (GRASS)
Software, Version 7.7. Open Source Geospatial Foundation. https://grass.osgeo.org/grass77/manuals/v.to.points.html
(2
June 2019). Hol´y, M., 1984. Vztahy mezi povrchov´ym odtokem a trans-
portem ˇzivin v povod´ı vod´arensk´ych n´adrˇz´ı (d´ılˇc´ı zpr´ava
v´yzkumn´eho ´ukolu VI 4 15/01 03/) (in czech). Technical report,
CTU in Prague, Prague. In the case of SMODERP2D model, the run-time is an issue, es-
pecially if multiple mid-scale hydrological basins in fine spatial
resolution grid computation needs to be undertaken. The code
parallelization is a common practice in cases where the reduc-
tion of run-time is convenient or even necessary, therefore the
existence of the TensorFlow-based branch; and although this
branch is still under development, the reduction of the computa-
tion costs is already reaching up to 40 per cent depending on the
data and architecture. This experiment also shows that the par-
allelized branch should not be used as the default one, but an ad Kalyanapu, A. J., Shankar, S., Pardyjak, E. R., Judi, D. R.,
Burian, S. J., 2011. Assessment of GPU computational en-
hancement to a 2D flood model. Environmental Modelling and
Software, 26(8), 1009–1016. Le, P. V., Kumar, P., Valocchi, A. J., Dang, H. V., 2015. GPU-
based high-performance computing for integrated surface-sub-
surface flow modeling. Environmental Modelling and Software,
73, 1–13. http://dx.doi.org/10.1016/j.envsoft.2015.07.015. This contribution has been peer-reviewed. https://doi.org/10.5194/isprs-archives-XLII-4-W14-143-2019 | © Authors 2019. CC BY 4.0 License. This contribution has been peer-reviewed. https://doi.org/10.5194/isprs-archives-XLII-4-W14-143-2019 | © Authors 2019. CC BY 4.0 License. p
https://doi.org/10.5194/isprs-archives-XLII-4-W14-143-2019 | © Authors 2019. CC BY 4.0 License. 3.
CONCLUSION 148 The International Archives of the Photogrammetry, Remote Sensing and Spatial Information Sciences, Volume XLII-4/W14, 2019
FOSS4G 2019 – Academic Track, 26–30 August 2019, Bucharest, Romania This contribution has been peer-reviewed. This contribution has been peer-reviewed.
htt
//d i
/10 5194/i
hi
XLII 4 W14 143 2019 | © A th
2019 CC BY 4 0 Li Molnar, D., Julien, P., 2000. Grid-size effects on surface runoff
modeling. Journal of Hydrologic Engineering, 5(1), 8–16. Molnar, D., Julien, P., 2000. Grid-size effects on surface runoff
modeling. Journal of Hydrologic Engineering, 5(1), 8–16. Neteler, M., Bowman, M., Landa, M., Metz, M., 2012. GRASS
GIS: a multi-purpose Open Source GIS. Environmental Mod-
elling & Software, 31, 124–130. O’Callaghan, J. F., Mark, D. M., 1984. The extraction of
drainage networks from digital elevation data. Computer Vision,
Graphics, and Image Processing, 28(3), 323 - 344. Philip, J., 1957. The theory of infiltration: 1. The infiltration
equation and its solution. Soil science, 83(5), 345–358. Puget,
J.,
2015. How
to
quickly
compute
the
man-
delbrot
set
in
python. IBM
Community
Blog. https://www.ibm.com/developerworks/community/blogs/jfp/en-
try/How To Compute Mandelbrodt Set Quickly?lang=en
(4
June 2019). Renard, K. G., Foster, G. R., Weesies, G., McCool, D., Yo-
der, D. et al., 1997. Predicting soil erosion by water: a guide
to conservation planning with the Revised Universal Soil Loss
Equation (RUSLE). 703, United States Department of Agricul-
ture Washington, DC. SMODERP2D
Development
Team,
2019. SMOD-
ERP2D
GitHub
repository. https://github.com/storm-fsv-
cvut/smoderp2d (2 June 2019). Vivoni, E. R., Mascaro, G., Mniszewski, S., Fasel, P., Springer,
E. P., Ivanov, V. Y., Bras, R. L., 2011. Real-world hydrologic
assessment of a fully-distributed hydrological model in a par-
allel computing environment. Journal of Hydrology, 409(1-2),
483–496. http://dx.doi.org/10.1016/j.jhydrol.2011.08.053. Wischmeier, W. H., Smith, D. D. et al., 1978. Predicting rain-
fall erosion losses-a guide to conservation planning. Predicting
rainfall erosion losses-a guide to conservation planning. Zambelli, P., Gebbert, S., Ciolli, M., 2013. Pygrass:
An
Object Oriented Python Application Programming Inter-
face (API) for Geographic Resources Analysis Support Sys-
tem (GRASS) Geographic Information System (GIS). IS-
PRS International Journal of Geo-Information, 2(1), 201–219. https://www.mdpi.com/2220-9964/2/1/201. Zhang, W., Cundy, T. W., 1989. Modeling of two-dimensional
overland flow. Water Resources Research, 25(9), 2019–2035. 149
|
https://openalex.org/W2584060087
|
https://journal.uokufa.edu.iq/index.php/kjvs/article/download/4019/3675
|
English
| null |
Resistance to cecal coccidiosis following sonicated oocysts immunization of Eimeri tenella in broilers
|
Kufa journal for veterinary medical sciences
| 2,015
|
cc-by
| 3,373
|
Kufa Journal For Veterinary Medical Sciences Vol. (6) No. (1) 2015 Kufa Journal For Veterinary Medical Sciences Vol. (6) No. (1) 2015 2015 Kufa Journa nal For Veterinary Medical Sciences
Vol (6)
No (1) Resistance to cecal coccidiosis following sonicated oocysts
immunization of Eimeri tenella in broilers Awse. M. Hussien** Latif I. Kadhim* *Pathology and Poultry Diseases Dept.-Coll of Vet. Med- Baghdad University
**Pathology and Poultry Diseases Dept.-Coll of Vet. Med- Diyala University *Pathology and Poultry Diseases Dept.-Coll of Vet. Med- Baghdad University
**Pathology and Poultry Diseases Dept.-Coll of Vet. Med- Diyala University Abstract Seventy five broiler chicks were used to determine the immunization of in ovo
inoculation or intramusclular (I/M) injection of sonicated oocyts (SO) antigen against
E. tenella manifestation. The chicks were divided in to 3 equal groups compromising
of 25 chicks each. The 1st group inoculated in ovo with SO of E. tenella and repeated
by I/M injection at 21 days old. The 2nd group injected I/M with SO at day old and
repeated at 21 days old, The birds of the 3rd group remains as control. All birds were
challenge by 50,000 sporulated oocysts of E. tenella at day 28. No significant (P >
0.05) group difference was detected between the immunized and non-immunized
groups for mean, WBC count, H/L ratio, lesion score, mortality and oocysts shedding
until E. tenella challenge at day 28. After challenge test the in ovo and I/M
immunized groups showed a great protective immunity against E. tenella infection
documented by significantly (P < 0.05) reduced mortality, lesion score and decreased
fecal oocyst shedding, compared with non-immunized group. It is concluded that in
ovo and I/M immuinzation of sonicated oocyst stimulates a remarkable protection in
broilers following E. tenella infection comparing with non-immunized groups. There
was no significance difference between the two procedures of immunization. Key words: E. tenella, immunity, sonicated oocyts, broilers g
p
Key words: E. tenella, immunity, sonicated oocyts, broilers المقاومة ضد االصابة االعورية لطفيلي
االيميريا تينيال
بعد التمنيع بالبيوض المكسرة في دجاج
اللحم
**لطيف ابراهيم كاظم* اوس المنتصر حسين
*فرع االمراض وامراض الدواجن-كلية الطب البيطري- جامعة بغداد
** فرع االمراض وامراض الدواجن-كلية الطب البيطري- جامعة ديالى المقاومة ضد االصابة االعورية لطفيلي
االيميريا تينيال
بعد التمنيع بالبيوض المكسرة في دجاج
اللحم
**لطيف ابراهيم كاظم* اوس المنتصر حسين
*فرع االمراض وامراض الدواجن-كلية الطب البيطري- جامعة بغداد
** فرع االمراض وامراض الدواجن-كلية الطب البيطري- جامعة ديالى Birds and management After cleaning and disinfection, one-
day-old, broiler chicks of the “Cobb
strain” were purchased from a local
hatchery. Upon arrival, the chicks were
raised
according
to
routine
management practice as outlined by
the
National
Research
Council
requirements
(8). All
nutrients
including water were supplied ad
libitum to meet the NRC (8). Vaccination is an alternative option for
coccidiosis control (5). Compared with
virulent or attenuated live vaccine,
recombinant
protein
vaccine
can
induce good antibody response and has
more efficiency to protect birds against
challenge of Eimeria oocysts (5). Parental inoculation of dead antigen is
although
capable
of
stimulating
circulating
antibodies
against
coccidiosis antigen (6). In spite of
these limitations, vaccination remains الخالصة: ا
أجريت هذه الدراسة على خمسة وسبعين فرخا من افراخ اللحم للتمنيع ضد اإلصابة بطفيلي االيميريا
تينيال باستخدام مستضد أكياس البيض المكسرة لطفيلي االيميريا تيني ال عن طريق الحقن بالبيض
والعضلة. قسمت
هذه األفراخ إلى3
مجاميع متساوية تحتوي كل مجموعة على52
فرخاً, حقنت المجموعة األولى بالبيض
ة
بعمر
81
يوماً من عمر الجنين وأعيد حقنها بعمر21
.يوماً بعد الفقس بأكياس البيض المكسرة لطفيلي االيميريا تينيال
المجموعة الثا
نية حقنت بالعضلة بعمر يوم و احد, وأعيد حقنها بعمر21
يوماً بمستضد أكياس البيض المكسرة 94 2015 Kufa Journal For Veterinary Medical Sciences Vol. (6) No. (1) لطفيلي االيميريا تينيال. المجموعة الثالثة تركت كمجموعة سيطرة. تم إجراء اختبار التحدي لكل الطيور بإعطائها
25,555
كيس بيض متبوغ لطفيلي االيميريا تينيال بعمر51
.ًيوما
لم تالحظ أي فروقات معنوية(P > 0.05)
بين المجاميع الممنعة والغير ممنعة من ناحية
معدل
المعايير الدموية
,المنتخبة
واالفات العيانية , ومعدل الهالك ومعدل طرح أكياس البيض في البراز حتى اجراء اختبار التحدي بعمر
51
.ًيوما
بعد إجراء اختبار التحدي أظهرت المجاميع الممنعة عن طريق الحقن بالبيضة والحقن بالعضلة
مستويات حماية عالية ضد ألخمج بطفيلي االيميريا تينيال وسجلت اختالفا معنوياً عند(P > 0.05)
من ناحية
انخفاض عدد الهالكات و اآلفات العيانية, و انخفاض
معدل أكياس البيض المطروحة مقارنةً مع المج موعة غير
.الممنعة
من هذا يمكن االستنتاج بأن التمنيع بواسطة الحقن بالبيض والحقن بالعضلة بأكياس البيض المكسرة
حفزت المناعة ووفرت حماية يمكن مالحظتها ضد الخمج بااليميريا تينيال مقارنةً مع المجاميع غير الممنعة. ولم
يكن هناك أي فرق معنوي بين طري .قتي التمنيع بواسطة الحقن بالبيض أو الحقن بالعضلة لطفيلي االيميريا تينيال. المجموعة الثالثة تركت كمجموعة سيطرة. تم إجراء اختبار التحدي لكل الطيور بإعطائها
25,555
كيس بيض متبوغ لطفيلي االيميريا تينيال بعمر51
.ًيوما
لم تالحظ أي فروقات معنوية(P > 0.05)
بين المجاميع الممنعة والغير ممنعة من ناحية
معدل
المعايير الدموية
,المنتخبة
واالفات العيانية , ومعدل الهالك ومعدل طرح أكياس البيض في البراز حتى اجراء اختبار التحدي بعمر
51
.ًيوما
بعد إجراء اختبار التحدي أظهرت المجاميع الممنعة عن طريق الحقن بالبيضة والحقن بالعضلة
مستويات حماية عالية ضد ألخمج بطفيلي االيميريا تينيال وسجلت اختالفا معنوياً عند(P > 0.05)
من ناحية
انخفاض عدد الهالكات و اآلفات العيانية, و انخفاض
معدل أكياس البيض المطروحة مقارنةً مع المج موعة غير
.الممنعة
من هذا يمكن االستنتاج بأن التمنيع بواسطة الحقن بالبيض والحقن بالعضلة بأكياس البيض المكسرة
حفزت المناعة ووفرت حماية يمكن مالحظتها ضد الخمج بااليميريا تينيال مقارنةً مع المجاميع غير الممنعة. ولم
يكن هناك أي فرق معنوي بين طري .قتي التمنيع بواسطة الحقن بالبيض أو الحقن بالعضلة Materials and Methods Experiment was done in the poultry
house
of
Pathology
and
Poultry
Diseases
Department,
College
of
Veterinary
Medicine,
Baghdad
University. At present, prophylactic chemotherapy
is still the main strategy of controlling
coccidiosis, but their extensive use
over the past 40 years has resulted in
the development of drug resistance by
these parasites (3). Due to food safety
concerns and the cost of new drug
development, recent emphasis has
centered on elicitation of protective
immune response to parasite infection
by development and effective use of
live or inactivated parasite vaccines
(4). Introduction Coccidiosis
is
an
economically
important
disease
that
seriously
impairs the feed utilization and growth
of chickens due to intestinal infection
with the protozoa Eimeria (1). Seven
species of Eimeria are generally
accepted to be causative agents of
avian coccidiosis, E. tenella found to
be the most prevalent and pathogenic
species throughout the world (2). the most efficient means of preventing
disease and reducing economic losses
(7). The purpose of the present study
was
to
protect
chicken
against
coccidiosis by immunized it with
whole
sonicated
oocyst
trial
in
alleviating E. tenella infection. Lesion scores
f
h Lesion scores
Means of the lesion scores of the
different
treatment
groups
were
determined. On days 28, 33, 34 and 35
five birds per group were randomly
selected and sacrificed by slaughtering
for scoring of lesions from cecal
coccidia. Lesions were scored in the
ceca on a scale from 1 to 4 with 4
being the most severe (15). Experimental Design A total of seventy five newly hatched
commercial
broiler
chicks
“Cobb
strain” was purchased from a local
hatchery. Upon arrival, the chicks were
divided randomly into three equal
groups of 25 chicks each, 1st group
chicks were inoculated in ovo with 0.2
ml of sonicated oocyst at day 18
embryonated eggs and repeated by I/M
injection at 21 days old. The 2nd group
chicks were receives 0.2 ml of
sonicated oocyst by I/M injection at
day one and repeated by same route at
21 days old. The 3rd group chicks
remain as control. All groups were
feed a basal diet without anticoccidial
drugs and challenged by 50,000 viable
sporulated oocysts of E. tenella at day
28 (11). Determination of oocysts shedding
The oocysts output was assessed using
a haemocytometer chamber method
which was documented by (16; 17). Litter was collected from several
representative areas of the chicken
house for oocyst counts at 10 days post
infection. The samples are mixed
together and 5 g of material are
weighed, and purification, flotation and
a haemocytometer counting was done. Total oocyst output= total oocyst count
per gram x weight of feces. Results The mean total WBCs count of broilers
during the experimental period is
summarized in Table 1. No significant
differences were shown between all
experimental groups at day 28. Then
after,
statistically
significant
differences were indicated in the
immunized groups (P < 0.05) when
compared with non-immunized group
at days 33, 34 and 35. At that time the
vaccinated ones had the low values. Sonicated
antigen
and
vaccine
preparation Field isolate of a highly virulent strain
of E. tenella was obtained from a
broiler farm with an outbreak of
bloody diarrhea with pathognomonic
cecal lesion and high mortality was
used
in
this
study
for
oocysts
collection. After oocysts collection in
potassium dichromate solution (% 2.5)
and sporulatation, the oocysts were 05 Kufa Journal For Veterinary Medical Sciences Vol. (6) No. (1) 2015 (EDTA), WBC were counted using the
haemocytometer (13), and heterophil
to lymphocyte ratio were determined
by counting encountered elements on
blood smears under the microscope
according to Hawk (14). washed 3 times by physiological saline
solution (pH 7.2) at 3000 rpm for 10
minutes each and concentrated to
5000-6000
per
ml
using
the
hemocytometer
(9). The
washed
sporulated oocysts were subjected to
ultra-sonication
by
Soniprep150
(SONY Company) for 2 by 30 seconds
in jacketed vessel with cool water. The
inactivated vaccine was prepared from
sonicated suspension by treating with
0.3
percent
formalin
(33%
formaldehyde) for 96 hours at 37°C
(10; 6) and stored at 4°C until use. Performance parameters
Morbidity and mortality rate The morbidity and mortality monitored
by recording the clinical signs and
dead chicks from 5th day to 8th day
after challenge infection with E. tenella
(12). Effects on selected haemograms Effects on selected haemograms
Blood (3 ml) was collected from the
birds from the jugular vein into 05 Kufa Journal For Veterinary Medical Sciences Vol. (6) No. (1) 2015 Table: 1 the WBCs counts (103/mm3) of broilers during the experimental period
(mean±SD) Table: 1 the WBCs counts (103/mm3) of broilers during the experimental period
(mean±SD)
Groups
Before challenge
After challenge
d 28
d 33
d 34
d 35
Inovo
20.48 ± 0.509 a
20.79 ± 0.750 b
20.95 ± 0.840 b
21.42 ± 0.916 a
I/M
21.48 ± 1.377 a
20.88 ± 0.993 b
20.99 ± 0.809 b
21.81 ± 0.777 a
Control
20.39 ± 1.042 a
22.95 ± 1.375 a
22.74 ± 0.890 a
22.58 ± 0.891 a
a, b Values bearing similar superscript between column do not differ at (P < 0.05). Similarly, an increasing pattern of H/L ratio was seen in non-immunized group at
days 33, 34 and 35 (Table 2). That of the immunized groups remained the lowest H/L
ratio (P < 0.05) at days 33, 34 and 35. y
le: 2 the H/L ratio of broilers during the experimental period (mean±SD) Table: 2 the H/L ratio of broilers during the experimental period (mean±SD)
Groups
Before challenge
After challenge
d 28
d 33
d 34
d 35
Inovo
0.373 ± 0.046 a
0.412 ± 0.044 b 0.404 ± 0.052 b
0.394 ± 0.054 b
I/M
0.401 ± 0.070 a
0.409 ± 0.032 b 0.406 ± 0.032 b
0.404 ± 0.032 b
Control
0.402 ± 0.059 a
0.554 ± 0.049 a 0.502 ± 0.048 a
0.509 ± 0.048 a
a, b Values bearing similar superscript between column do not differ at (P < 0.05). a, b Values bearing similar superscript between column do not differ at (P < 0.05). The lesions score of broilers during the experimental period (mean±SD) are illustrated
in table 3. Lesion scores were zero for the all groups before challenge. After
challenge, the immunized groups have the lowest mean lesion compared with non-
immunized control group which was had the highest mean lesion scores with an
average of 3.20. average of 3.20. Table: 3 the lesions score of broilers during the experimental period (mean±SD)
Groups
Before challenge
After challenge
d 28
d 35
Inovo
0
1.00±0.707 b
I/M
0
0.80±0.447 b
Control
0
3.20±0.447 a
a, b Values bearing similar superscript between column do not differ at (P < 0.05). Kufa Journal For Veterinary Medical Sciences Vol. (6) No. (1) Kufa Journal For Veterinary Medical Sciences Vol. (6) No. (1) 2015 The results of oocyst output of broilers during the experimental period are
summarized in table 5. The immunized groups shed the lowest oocysts (0.051xl06,
0.055xl06) of any of the infected groups. Nevertheless, the control group had the
highest means with an estimate of 1.930x106 oocysts shedding. g
y
g
Table: 5 the oocyst output (x106) of broilers during the experimental period
(mean±SD)
Groups
d 28
d 38
Inovo
0
0.051±0.016 b
I/M
0
0.055±0.009 b
Control
0
1.930±0.155 a
a, b Values bearing similar superscript between column do not differ at (P < 0.05). g
y
g
Table: 5 the oocyst output (x106) of broilers during the experimental period
(mean±SD) H/L ratio from normal in the non-
immunized
groups,
this
increase
corresponds to periods of severe tissue
destruction to the heavy infestation of
E. tenella (22). Discussions The effectiveness of vaccination in the
generation of immunity and protection
against subsequent Eimeria infection
has been well documented (4). The in
ovo inoculation at day 18 embryonated
eggs and I/M at 1 day old injection
proved their ability to stimulating the
immune system ( 18; 19). The main
objectives of this research were to
assess whether the sonicated oocysts
antigen could reduce the negative
effects to mean protection rate, cecal
lesions and oocyst output and mortality
by
stimulating
effective
immune
response associated with virulent E. tenella challenged in broiler chickens. Our results revealed that there were no
statistically differences were detected
in the immunized and non-immunized
groups for mean, WBC count, H/L
ratio, lesion score, mortality and
oocysts shedding until E. tenella
challenge at day 28, this proved that
the dose of antigen not effected on the
chicks. In this study immunization with the E. tenella sonicated oocyts stimulated
protective
immunity
against
experimental challenge infection as
indicated by reduced lesion scores
among immunized groups compared
with non-immunized groups (cecal
lesions in the 2 to 3 range), vaccination
against E. tenella reduced the cecal
distraction (4). The mortality rate in
non-immunized groups showed high
levels comparing with the immunized
groups which proved high protection
against the E. tenella. Our results revealed that there were no
statistically differences were detected
in the immunized and non-immunized
groups for mean, WBC count, H/L
ratio, lesion score, mortality and
oocysts shedding until E. tenella
challenge at day 28, this proved that
the dose of antigen not effected on the
chicks. g
In general protective immunity has
normally
been
measured
through
vaccination by reduction in lesion
scores, cessation in total oocyst output
(23). We
can
conclude
that
immunization of broilers chicks with a
sonicated oocyst is an effective tool for
generation of protective immunity to
field strain of E. tenella following
challenged of poultry with this antigen
increased this protection and eliminate
the disease by prevent the clinical
signs, gross lesions and mortality rate. In general protective immunity has
normally
been
measured
through
vaccination by reduction in lesion
scores, cessation in total oocyst output
(23). We
can
conclude
that
immunization of broilers chicks with a
sonicated oocyst is an effective tool for
generation of protective immunity to
field strain of E. Discussions tenella following
challenged of poultry with this antigen
increased this protection and eliminate
the disease by prevent the clinical
signs, gross lesions and mortality rate. White blood cells are the effectors cells
of immune responses and have been
assessed
in
many
immunological
studies of avian (20). The normal range
limits
of
selected
haemogram
parameters
values
(21)
in
the
immunized groups signified possible
event of a usual situation compared
with the deviation of WBC count and Effects on selected haemograms g
Table: 3 the lesions score of broilers during the experimental period (mean±SD) a, b Values bearing similar superscript between column do not differ at (P < 0.05). In the cross sectional study, noticeable signs of morbidity were observed as
depression, ruffled feather, diarrhea and/or blood mixed droppings were recorded as
clinical cases at day 5 post infection and peaked on days 6 and 7. The result of clinical
infection was observed in non-immunized group (control). One bird died for each in
ovo and I/M groups, then six birds was died from control group (table 4). g
p
g
p (
)
Table: 4 the mortality of broilers during the experimental period (mean±SD) Groups
Number of challenge at d 28
After challenge till d 35
Percentage
Inovo
15
1
6.66
I/M
15
1
6.66
Control
15
6
40.00 05 References: 1. Allen, D.C. and Fetterer, R.H. (2002). Recent
advances
in 05 2015 Kufa Journal For Veterinary Medical Sciences Vol. (6) No. (1) biology and immunobiology of
Emeira Species and in diagnosis
and control of infection with
coccidiosis
of
poultry. Clin. Microbiol. Rev. 15:58-65. 9. Davis, L.R. (1973). Techniques. In
The
Coccidia,
ed. D. M. Hammond and P. L. Long, 411–
58. Baltimore, MD: Univ. Park
Press. 10. Fu, H.M. and Lee Y.C. (1976):
Immunological
studies
on
chemically attenuated oocysts of
chicken caecal coccidiosis. Journal
of Chinese Society of Vet. Sci. 2:
51–55. 2. Haug, A., Gjevre, A.G., Thebo, P.,
Mattsson, J.G. and Kaldhusdal, M. (2008). Coccidial infections in
commercial
broilers:
epidemiological
aspects
and
comparison of Eimeria species
identification by morphometric
and polymerase chain reaction
techniques. Avian Pathol. 37:161–
170. 11. Reid, W.M. (1978). Coccidiosis. In Diseases of Poultry, 7th ed., ed. M. S. Hofstad, B.W. Calnek, C. F. Helmboldt,W. M. Reid, and H. W. Yoder, Jr. 784-815. Ames, IA:
Iowa State Univ. Press. 3. Kitandu, A. and Juranovà, R. (2006). Progress
in
Control
Measures for Chicken Coccidiosis
ACTA VET. BRNO, 75: 265–276. 12. Edward, C.M., Williams, C.C. and
Cuckler,
A.C. (1968). Development
to
quinoline
coccidiostats
under
field
and
laboratory conditions. J. Parasitol. 45: 1190-1193. 4. Dalloul, R.A. and Lillehoj, H.S. (2005). Recent
advances
in
immunomodulation
and
vaccination
strategies
against
coccidiosis. Avian Dis. 49: 1-8. 13. Jain,
C.N. (1986). Schalm’s
veterinary haematology, 4th ed. Lea and Febiger, Philadelphia, p. 42. 5. Geriletu, Xu, L., Xurihu, and Li,
X. (2011). Vaccination of chickens
with DNA vaccine expressing
Eimeria tenella MZ5-7 against
coccidiosis Vet. Parasitol.177: 6–
12. 14. Hawk, P.B., Oscar, B.L. and
Summerson, W. (1965). Hawk's
physiological chemistry. 14th Ed. London
Journal;
and
A. Churchill.L.T.D.401. 6. Bahram, A.M. and Bahrami, A. (2006). Immune
response
of
chicken
to
an
experimental
sonicated
coccidian
oocyst
vaccine. Archives
of
Razi
Institute, 61: 43-48. 15. Johnson, J.K. and Reid, W.M. (1970). Anticoccidial drugs: lesion
scoring techniques in battery and
floor-pen
experiments
with
chickens. Exp. Parasitol. 28: 30-
36. 7. Sharman,
P.A.,
Smith,
N.C.,
Wallach, M.G. and Katrib, M. (2010). Chasing the golden egg:
Vaccination
against
poultry
coccidiosis. Parasite Immunol. 32:
590-598. 16. Long, Μ.Α., Handwerger, Β. S.,
Arnos, D.Β. and Yunis, Ε.J. (1976). The genetics of cell
mediated
lympholysis. J. Immuno. 117: 2092-2099. 8. National
Research
Council. (1994). Nutrient Requirements for
Poultry. 9th
(ed),
pp
3-34. Washington: National Academy
Press. 8. National
Research
Council. (1994). References: Nutrient Requirements for
Poultry. 9th
(ed),
pp
3-34. Washington: National Academy
Press. 17. Hodgson,
J.N. (1970). Coccidiosis:
oocyst
counting
technique
for
coccidiostat
evaluation. Exp. Parasitol. 28: 99-
102. 09 Kufa Journal For Veterinary Medical Sciences Vol. (6) No. (1) 2015 18. Long, P.L. (1984). Coccidiosis
control: past present and future. British poult. Sci. 25: 3-18. 19. Weber, F.H. and Evans. N.A. (2003). Immunization of broiler
chicks by in ovo injection of
Eimeria
tenella
sporozoites,
sporocysts, or oocysts. Poult. Sci. 82: 1701–1707. 20. Grasman, K.A.)2002(. Assessing
immunological
function
in
toxicological studies of avian
wildlife. Integr. Comp. Biol. 42:
34–42. 21. Bartholomew, A., Latshaw, D. and
Swane, D. (1998). Changes in
blood chemistry, hematology, and
histology
caused
by
a
selenium/vitamin E deficiency and
recovery in chicks. Biol. Trace
Element Res. 62: 7–16. 22. Kogut, M.H., Gore, T.C. and
Long,
P.L. (1984). Eimeria
tenella,
Emeria
necatrix,
and
Eimeria adenoeides: Peripheral
blood
leukocyte
response
of
chickens and turkeys to strains
adapted to the turkey embryo. Exp. Parasitol. 58: 63-71. 23. De
Gussem,
M. (2007). Coccidiosis in poultry: Review on
diagnosis, control, prevention and
interaction with overall gut health. Pages 253–261 in Proc. 16th Eur. Symp. on Poult. Nutr. World’s
Poultry
Science
Association,
Beekbergen, the Netherlands. 00
|
https://openalex.org/W2590000561
|
https://repositorio.unican.es/xmlui/bitstream/10902/1967/3/MeasurementTopPair.pdf
|
English
| null |
Measurement of the top pair production cross section in the dilepton decay channel in<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" display="inline"><mml:mi>p</mml:mi><mml:mover accent="true"><mml:mi>p</mml:mi><mml:mo>¯</mml:mo></mml:mover></mml:math>collisions at<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" display="inline"><mml:msqrt><mml:mi>s</mml:mi></mml:msqrt><mml:mo>=</mml:mo><mml:mn>1.96</mml:mn><mml:mtext> </mml:mtext><mml:mtext> </mml:mtext><mml:mi>TeV</mml:mi></…
|
Physical review. D. Particles, fields, gravitation, and cosmology/Physical review. D, Particles, fields, gravitation, and cosmology
| 2,010
|
cc-by
| 20,478
|
PHYSICAL REVIEW D 82, 052002 (2010) PHYSICAL REVIEW D 82, 052002 (2010) PHYSICAL REVIEW D 82, 052002 (2010) in p p collisions at
ffiffiffis
p ¼ 1:96 TeV
T. Aaltonen,22 B. A´ lvarez Gonza´lez,10,x S. Amerio,42a D. Amidei,33 A. Anastassov,37 A. Annovi,18 J. Antos,13
G. Apollinari,16 J. A. Appel,16 A. Apresyan,47 T. Arisawa,56 A. Artikov,14 J. Asaadi,52 W. Ashmanskas,16 B. Auerbach,59
A. Aurisano,52 F. Azfar,41 W. Badgett,16 A. Barbaro-Galtieri,27 V. E. Barnes,47 B. A. Barnett,24 P. Barria,45a,45c P. Bartos,13
M. Bauce,42a,42b G. Bauer,31 F. Bedeschi,45a D. Beecher,29 S. Behari,24 G. Bellettini,45a,45b J. Bellinger,58 D. Benjamin,15
A. Beretvas,16 A. Bhatti,49 M. Binkley,16,a D. Bisello,42a,42b I. Bizjak,29,dd K. R. Bland,5 C. Blocker,7 B. Blumenfeld,24
A. Bocci,15 A. Bodek,48 D. Bortoletto,47 J. Boudreau,46 A. Boveia,12 B. Brau,16,c L. Brigliadori,6a,6b A. Brisuda,13
C. Bromberg,34 E. Brucken,22 M. Bucciantonio,45a,45b J. Budagov,14 H. S. Budd,48 S. Budd,23 K. Burkett,16
G. Busetto,42a,42b P. Bussey,20 A. Buzatu,32 S. Cabrera,15,z C. Calancha,30 S. Camarda,4 M. Campanelli,34 M. Campbell,33
F. Canelli,12,16 A. Canepa,44 B. Carls,23 D. Carlsmith,58 R. Carosi,45a S. Carrillo,17,m S. Carron,16 B. Casal,10 M. Casarsa,16
A. Castro,6a,6b P. Catastini,16 D. Cauz,53a V. Cavaliere,45a,45c M. Cavalli-Sforza,4 A. Cerri,27,h L. Cerrito,29,s Y. C. Chen,1
M. Chertok,8 G. Chiarelli,45a G. Chlachidze,16 F. Chlebana,16 K. Cho,26 D. Chokheli,14 J. P. Chou,21 W. H. Chung,58
Y. S. Chung,48 C. I. Ciobanu,43 M. A. Ciocci,45a,45c A. Clark,19 D. Clark,7 G. Compostella,42a,42b M. E. Convery,16
J. Conway,8 M. Corbo,43 M. Cordelli,18 C. A. Cox,8 D. J. Cox,8 F. Crescioli,45a,45b C. Cuenca Almenar,59 J. Cuevas,10,x
R. Culbertson,16 D. Dagenhart,16 N. d’Ascenzo,43,v M. Datta,16 P. de Barbaro,48 S. De Cecco,50a G. De Lorenzo,4
M. Dell’Orso,45a,45b C. Deluca,4 L. Demortier,49 J. Deng,15,e M. Deninno,6a F. Devoto,22 M. d’Errico,42a,42b
A. Di Canto,45a,45b B. Di Ruzza,45a J. R. Dittmann,5 M. D’Onofrio,28 S. Donati,45a,45b P. Dong,16 T. Dorigo,42a K. Ebina,56
A. Elagin,52 A. Eppig,33 R. Erbacher,8 D. Errede,23 S. Errede,23 N. Ershaidat,43,cc R. Eusebi,52 H. C. Fang,27
S. Farrington,41 M. Feindt,25 J. P. Fernandez,30 C. Ferrazza,45a,45d R. Field,17 G. Flanagan,47,t R. Forrest,8 M. J. Frank,5
M. Franklin,21 J. C. Freeman,16 I. Furic,17 M. Gallinaro,49 J. Galyardt,11 J. E. Garcia,19 A. F. Garfinkel,47 P. Garosi,45a,45c
H. Gerberich,23 E. Gerchtein,16 S. Giagu,50a,50b V. Giakoumopoulou,3 P. Giannetti,45a K. Gibson,46 C. M. Ginsburg,16
N. Giokaris,3 P. Giromini,18 M. Giunta,45a G. Giurgiu,24 V. Glagolev,14 D. Glenzinski,16 M. Gold,36 D. Goldin,52
N. Goldschmidt,17 A. Golossanov,16 G. Gomez,10 G. Gomez-Ceballos,31 M. Goncharov,31 O. Gonza´lez,30 I. Gorelov,36
A. T. Goshaw,15 K. Goulianos,49 A. Gresele,42a S. Grinstein,4 C. Grosso-Pilcher,12 R. C. Group,16 J. Guimaraes da Costa,21
Z. Gunay-Unalan,34 C. Haber,27 S. R. Hahn,16 E. Halkiadakis,51 A. Hamaguchi,40 J. Y. Han,48 F. Happacher,18 K. Hara,54
D. Hare,51 M. Hare,55 R. F. Harr,57 K. Hatakeyama,5 C. Hays,41 M. Heck,25 J. Heinrich,44 M. Herndon,58 S. Hewamanage,5
D. Hidas,51 A. Hocker,16 W. Hopkins,16,i D. Horn,25 S. Hou,1 R. E. Hughes,38 M. Hurwitz,12 U. Husemann,59 N. Hussain,32
M. Hussein,34 J. Huston,34 G. Introzzi,45a M. Iori,50a,50b A. Ivanov,8,q E. James,16 D. Jang,11 B. Jayatilaka,15 E. J. Jeon,26
M. K. Jha,6a S. Jindariani,16 W. Johnson,8 M. Jones,47 K. K. Joo,26 S. Y. Jun,11 T. R. Junk,16 T. Kamon,52 P. E. Karchin,57
Y. Kato,40,p W. Ketchum,12 J. Keung,44 V. Khotilovich,52 B. Kilminster,16 D. H. Kim,26 H. S. Kim,26 H. W. Kim,26
J. E. Kim,26 M. J. Kim,18 S. B. Kim,26 S. H. Kim,54 Y. K. Kim,12 N. Kimura,56 S. Klimenko,17 K. Kondo,56 D. J. Kong,26
J. Konigsberg,17 A. Korytov,17 A. V. Kotwal,15 M. Kreps,25 J. Kroll,44 D. Krop,12 N. Krumnack,5,n M. Kruse,15
V. Krutelyov,52,f T. Kuhr,25 M. Kurata,54 S. Kwang,12 A. T. Laasanen,47 S. Lami,45a S. Lammel,16 M. Lancaster,29
R. L. Lander,8 K. Lannon,38,w A. Lath,51 G. Latino,45a,45c I. Lazzizzera,42a T. LeCompte,2 E. Lee,52 H. S. Lee,12 J. S. Lee,26
S. W. Lee,52,y S. Leo,45a,45b S. Leone,45a J. D. Lewis,16 C.-J. Lin,27 J. Linacre,41 M. Lindgren,16 E. Lipeles,44 A. Lister,19
D. O. Litvintsev,16 C. Liu,46 Q. Liu,47 T. Liu,16 S. Lockwitz,59 N. S. Lockyer,44 A. Loginov,59 D. Lucchesi,42a,42b
J. Lueck,25 P. Lujan,27 P. Lukens,16 G. Lungu,49 J. Lys,27 R. Lysak,13 R. Madrak,16 K. Maeshima,16 K. Makhoul,31
P. Maksimovic,24 S. Malik,49 G. Manca,28,d A. Manousakis-Katsikakis,3 F. Margaroli,47 C. Marino,25 M. Martı´nez,4
R. Martı´nez-Balları´n,30 P. Mastrandrea,50a M. Mathis,24 M. E. Mattson,57 P. Mazzanti,6a K. S. McFarland,48 P. McIntyre,52
R. McNulty,28,k A. Mehta,28 P. Mehtala,22 A. Menzione,45a C. Mesropian,49 T. Miao,16 D. Mietlicki,33 A. Mitra,1
G. Mitselmakher,17 H. Miyake,54 S. Moed,21 N. Moggi,6a M. N. Mondragon,16,m C. S. Moon,26 R. Moore,16
M. J. Morello,16 J. Morlock,25 P. Movilla Fernandez,16 A. Mukherjee,16 Th. Muller,25 P. Murat,16 M. Mussini,6a,6b
J. Nachtman,16,o Y. Nagai,54 J. Naganoma,56 I. Nakano,39 A. Napier,55 J. Nett,58 C. Neu,44,bb M. S. Neubauer,23
J. Nielsen,27,g L. Nodulman,2 O. Norniella,23 E. Nurse,29 L. Oakes,41 S. H. Oh,15 Y. D. Oh,26 I. Oksuzian,17 T. Okusawa,40
R. Orava,22 L. Ortolan,4 S. Pagan Griso,42a,42b C. Pagliarone,53a E. Palencia,10,h V. Papadimitriou,16 A. A. Paramonov,2
J. Patrick,16 G. Pauletta,53a,53b M. Paulini,11 C. Paus,31 D. E. Pellett,8 A. Penzo,53a T. J. Phillips,15 G. Piacentino,45a
E. Pianori,44 J. Pilot,38 K. Pitts,23 C. Plager,9 L. Pondrom,58 K. Potamianos,47 O. Poukhov,14,a F. Prokoshin,14,aa
A. Pronko,16 F. Ptohos,18,j E. Pueschel,11 G. Punzi,45a,45b J. Pursley,58 A. Rahaman,46 V. Ramakrishnan,58 N. Ranjan,47
I. Redondo,30 P. Renton,41 M. Rescigno,50a F. Rimondi,6a,6b L. Ristori,45a,16 A. Robson,20 T. Rodrigo,10 T. Rodriguez,44 Measurement of the top pair production cross section in the dilepton decay channel
in p p collisions at
ffiffiffis
p ¼ 1:96 TeV A. Castro,
P. Catastini,
D. Cauz,
V. Cavaliere,
M. Cavalli-Sforza, A. Cerri,
L. Cerrito,
Y. C. Chen,
M. Chertok,8 G. Chiarelli,45a G. Chlachidze,16 F. Chlebana,16 K. Cho,26 D. Chokheli,14 J. P. Chou,21 W. H. Chung,58
Y. S. Chung,48 C. I. Ciobanu,43 M. A. Ciocci,45a,45c A. Clark,19 D. Clark,7 G. Compostella,42a,42b M. E. Convery,16 g
pp g
S. Farrington,41 M. Feindt,25 J. P. Fernandez,30 C. Ferrazza,45a,45d R. Field,17 G. Flanagan,47, M. Franklin,21 J. C. Freeman,16 I. Furic,17 M. Gallinaro,49 J. Galyardt,11 J. E. Garcia,19 A. F. Garfinkel,47 P. Garosi,45a,45c H. Gerberich,23 E. Gerchtein,16 S. Giagu,50a,50b V. Giakoumopoulou,3 P. Giannetti,45a K. Gibson,46 C. M. Ginsburg,16
N. Giokaris,3 P. Giromini,18 M. Giunta,45a G. Giurgiu,24 V. Glagolev,14 D. Glenzinski,16 M. Gold,36 D. Goldin,52 A. T. Goshaw,15 K. Goulianos,49 A. Gresele,42a S. Grinstein,4 C. Grosso-Pilcher,12 R. C. Group,16 J. Guimaraes da Costa,21 D. Hare,51 M. Hare,55 R. F. Harr,57 K. Hatakeyama,5 C. Hays,41 M. Heck,25 J. Heinrich,44 M. Herndon,58 S. Hewamanage,5 M. Hussein,34 J. Huston,34 G. Introzzi,45a M. Iori,50a,50b A. Ivanov,8,q E. James,16 D. Jang,11 B. Jayatilaka,15 E. J. Jeon,26 g
g
p
J. Nielsen,27,g L. Nodulman,2 O. Norniella,23 E. Nurse,29 L. Oakes,41 S. H. Oh,15 Y. D. Oh,26 I. Oksuzian,17 T. Okusawa
22
4
42 42b
53
10 h
16 R. Orava,22 L. Ortolan,4 S. Pagan Griso,42a,42b C. Pagliarone,53a E. Palencia,10,h V. Papadimitriou,16 A. A. Paramonov,2
J. Patrick,16 G. Pauletta,53a,53b M. Paulini,11 C. Paus,31 D. E. Pellett,8 A. Penzo,53a T. J. Phillips,15 G. Piacentino,45a
E. Pianori,44 J. Pilot,38 K. Pitts,23 C. Plager,9 L. Pondrom,58 K. Potamianos,47 O. Poukhov,14,a F. Prokoshin,14,aa g
A. Pronko,16 F. Ptohos,18,j E. Pueschel,11 G. Punzi,45a,45b J. Pursley,58 A. Rahaman,46 V. Ramakrishnan,58 N. Ranjan,47 1550-7998=2010=82(5)=052002(20) 2010 The American Physical Society T. AALTONEN et al. Vidal,30 I. Vila,10 R. Vilar,10
M. Vogel,36 G. Volpi,45a,45b P. Wagner,44 R. L. Wagner,16 T. Wakisaka,40 R. Wallny,9 S. M. Wang,1 A. Warburton,32 S. Uozumi,
A. Varganov,
E. Vataga,
F. Vazquez,
G. Velev,
C. Vellidis, M. Vidal,
I. Vila,
R. Vilar,
M. Vogel,36 G. Volpi,45a,45b P. Wagner,44 R. L. Wagner,16 T. Wakisaka,40 R. Wallny,9 S. M. Wang,1 A. Warburton,32 ,
g
,
g ,
q
,
,
,
,
,
,
M. Vogel,36 G. Volpi,45a,45b P. Wagner,44 R. L. Wagner,16 T. Wakisaka,40 R. Wallny,9 S. M. Wang,1 A. Warburton,32
D. Waters,29 M. Weinberger,52 W. C. Wester III,16 B. Whitehouse,55 D. Whiteson,44,e A. B. Wicklund,2 E. Wicklund,16
S. Wilbur,12 F. Wick,25 H. H. Williams,44 J. S. Wilson,38 P. Wilson,16 B. L. Winer,38 P. Wittich,16,i S. Wolbers,16 H. Wolfe,38
T. Wright,33 X. Wu,19 Z. Wu,5 K. Yamamoto,40 J. Yamaoka,15 U. K. Yang,12,r Y. C. Yang,26 W.-M. Yao,27 G. P. Yeh,16
K. Yi,16,o J. Yoh,16 K. Yorita,56 T. Yoshida,40,l G. B. Yu,15 I. Yu,26 S. S. Yu,16 J. C. Yun,16 A. Zanetti,53a
15
6 6b g
p
g
g
y
g
D. Waters,29 M. Weinberger,52 W. C. Wester III,16 B. Whitehouse,55 D. Whiteson,44,e A. B. Wicklund,2 E. Wicklund,16
S. Wilbur,12 F. Wick,25 H. H. Williams,44 J. S. Wilson,38 P. Wilson,16 B. L. Winer,38 P. Wittich,16,i S. Wolbers,16 H. Wolfe,38
T. Wright,33 X. Wu,19 Z. Wu,5 K. Yamamoto,40 J. Yamaoka,15 U. K. Yang,12,r Y. C. Yang,26 W.-M. Yao,27 G. P. Yeh,16 D. Waters,
M. Weinberger,
W. C. Wester III,
B. Whitehouse,
D. Whiteson,
, A. B. Wicklund, E. Wicklund,
S. Wilbur,12 F. Wick,25 H. H. Williams,44 J. S. Wilson,38 P. Wilson,16 B. L. Winer,38 P. Wittich,16,i S. Wolbers,16 H. Wolfe,38 T. Wright,33 X. Wu,19 Z. Wu,5 K. Yamamoto,40 J. Yamaoka,15 U. K. Yang,12,r Y. C. Yang,26
K. Yi,16,o J. Yoh,16 K. Yorita,56 T. Yoshida,40,l G. B. Yu,15 I. Yu,26 S. S. Yu,16 J. C. Yu T. Wright,33 X. Wu,19 Z. Wu,5 K. Yamamoto,40 J. Yamaoka,15 U. K. Yang,12,r Y. C. Yang,26 W.-M. Yao,27 G. P. Yeh,16
K. Yi,16,o J. Yoh,16 K. Yorita,56 T. Yoshida,40,l G. B. Yu,15 I. Yu,26 S. S. Yu,16 J. C. Yun,16 A. Zanetti,53a g
g
g
K. Yi,16,o J. Yoh,16 K. Yorita,56 T. Yoshida,40,l G. B. Yu,15 I. Yu,26 S. S. Yu,16 J. C. Yun,16 A. Zanetti,53a Y. Zeng,15 and S. Zucchelli6a,6b 052002-1 052002-1 052002-1 T. AALTONEN et al. Tang,12 M. Tecchio,33 P. K. Teng,1 J. Thom,16,i J. Thome,11
G. A. Thompson,23 E. Thomson,44 P. Ttito-Guzma´n,30 S. Tkaczyk,16 D. Toback,52 S. Tokar,13 K. Tollefson,34 T. Tomura,54
D. Tonelli,16 S. Torre,18 D. Torretta,16 P. Totaro,53a,53b M. Trovato,45a,45d Y. Tu,44 N. Turini,45a,45c F. Ukegawa,54
S. Uozumi,26 A. Varganov,33 E. Vataga,45a,45d F. Va´zquez,17,m G. Velev,16 C. Vellidis,3 M. Vidal,30 I. Vila,10 R. Vilar,10
M. Vogel,36 G. Volpi,45a,45b P. Wagner,44 R. L. Wagner,16 T. Wakisaka,40 R. Wallny,9 S. M. Wang,1 A. Warburton,32
D. Waters,29 M. Weinberger,52 W. C. Wester III,16 B. Whitehouse,55 D. Whiteson,44,e A. B. Wicklund,2 E. Wicklund,16
S. Wilbur,12 F. Wick,25 H. H. Williams,44 J. S. Wilson,38 P. Wilson,16 B. L. Winer,38 P. Wittich,16,i S. Wolbers,16 H. Wolfe,38
T Wright 33 X Wu 19 Z Wu 5 K Yamamoto 40 J Yamaoka 15 U K Yang 12,r Y C Yang 26 W M Yao 27 G P Yeh 16 g
y
A. Sukhanov,17 I. Suslov,14 K. Takemasa,54 Y. Takeuchi,54 J. Tang,12 M. Tecchio,33 P. K. Teng,1 J. Thom,16,i J. Thome,11 A. Sukhanov,
I. Suslov,
K. Takemasa,
Y. Takeuchi,
J. Tang,
M. Tecchio,
P. K. Teng, J. Thom,
J. Thome,
G. A. Thompson,23 E. Thomson,44 P. Ttito-Guzma´n,30 S. Tkaczyk,16 D. Toback,52 S. Tokar,13 K. Tollefson,34 T. Tomura,54
D. Tonelli,16 S. Torre,18 D. Torretta,16 P. Totaro,53a,53b M. Trovato,45a,45d Y. Tu,44 N. Turini,45a,45c F. Ukegawa,54
S Uozumi 26 A Varganov 33 E Vataga 45a,45d F Va´zquez 17,m G Velev 16 C Vellidis 3 M Vidal 30 I Vila 10 R Vilar 10 ,
,
,
,
g,
,
g,
,
,
G. A. Thompson,23 E. Thomson,44 P. Ttito-Guzma´n,30 S. Tkaczyk,16 D. Toback,52 S. Tokar,13 K. Tollefson,34 T. Tomura,54
D. Tonelli,16 S. Torre,18 D. Torretta,16 P. Totaro,53a,53b M. Trovato,45a,45d Y. Tu,44 N. Turini,45a,45c F. Ukegawa,54 g
S. Uozumi,26 A. Varganov,33 E. Vataga,45a,45d F. Va´zquez,17,m G. Velev,16 C. Vellidis,3 M. Vidal,30 I. Vila,10 R. Vilar,10
M. Vogel,36 G. Volpi,45a,45b P. Wagner,44 R. L. Wagner,16 T. Wakisaka,40 R. Wallny,9 S. M. Wang,1 A. Warburton,32
D. Waters,29 M. Weinberger,52 W. C. Wester III,16 B. Whitehouse,55 D. Whiteson,44,e A. B. Wicklund,2 E. Wicklund,16
S. Wilbur,12 F. Wick,25 H. H. Williams,44 J. S. Wilson,38 P. Wilson,16 B. L. Winer,38 P. Wittich,16,i S. Wolbers,16 H. Wolfe,38
T. Wright,33 X. Wu,19 Z. Wu,5 K. Yamamoto,40 J. Yamaoka,15 U. K. Yang,12,r Y. C. Yang,26 W.-M. Yao,27 G. P. Yeh,16 S. Uozumi,26 A. Varganov,33 E. Vataga,45a,45d F. Va´zquez,17,m G. Velev,16 C. Vellidis,3 M. T. AALTONEN et al. T. AALTONEN et al. E. Rogers,23 S. Rolli,55 R. Roser,16 M. Rossi,53a F. Ruffini,45a,45c A. Ruiz,10 J. Russ,11 V. Rusu,16 A. Safonov,52
W. K. Sakumoto,48 L. Santi,53a,53b L. Sartori,45a K. Sato,54 V. Saveliev,43,v A. Savoy-Navarro,43 P. Schlabach,16
A. Schmidt,25 E. E. Schmidt,16 M. P. Schmidt,59,a M. Schmitt,37 T. Schwarz,8 L. Scodellaro,10 A. Scribano,45a,45c E. Rogers,
S. Rolli,
R. Roser,
M. Rossi,
F. Ruffini,
A. Ruiz,
J. Russ,
V. Rusu,
A. Safonov,
W. K. Sakumoto,48 L. Santi,53a,53b L. Sartori,45a K. Sato,54 V. Saveliev,43,v A. Savoy-Navarro,43 P. Schlabach,16
A S h idt 25 E E S h idt 16 M P S h idt 59 a M S h itt 37 T S h
8 L S
d ll
10 A S ib
45a 45c F. Scuri,45a A. Sedov,47 S. Seidel,36 Y. Seiya,40 A. Semenov,14 F. Sforza,45a,45b A. Sfyrla,23 S. Z. Shalhout,8 T. Shears,28
P. F. Shepard,46 M. Shimojima,54,u S. Shiraishi,12 M. Shochet,12 I. Shreyber,35 A. Simonenko,14 P. Sinervo,32
14 a
55
8
16
16
51
4
16 F. Scuri,45a A. Sedov,47 S. Seidel,36 Y. Seiya,40 A. Semenov,14 F. Sforza,45a,45b A. Sfyrla,23 S. Z. Shalhout,8 T. Shears,28
P. F. Shepard,46 M. Shimojima,54,u S. Shiraishi,12 M. Shochet,12 I. Shreyber,35 A. Simonenko,14 P. Sinervo,32
A. Sissakian,14,a K. Sliwa,55 J. R. Smith,8 F. D. Snider,16 A. Soha,16 S. Somalwar,51 V. Sorin,4 P. Squillacioti,16
59
20
32
32
37
36
33
54 P. F. Shepard,46 M. Shimojima,54,u S. Shiraishi,12 M. Shochet,12 I. Shreyber,35 A. Simonenko,14 P. Sinervo,32
A. Sissakian,14,a K. Sliwa,55 J. R. Smith,8 F. D. Snider,16 A. Soha,16 S. Somalwar,51 V. Sorin,4 P. Squillacioti,16
S
i ki59
S
i
20
S l
32 O S l
Chil
32
S
37
S
l
36 G
S
k
33
S d
54 P. F. Shepard,
M. Shimojima,
S. Shiraishi,
M. Shochet,
I. Shreyber,
A. Simonenko,
P. Sinervo,
A. Sissakian,14,a K. Sliwa,55 J. R. Smith,8 F. D. Snider,16 A. Soha,16 S. Somalwar,51 V. Sorin,4 P. Squillacioti,16
M St
it ki 59 R St D
i
20 B St l
32 O St l
Chilt
32 D St
t
37 J St
l
36 G L St
k
33 Y S d
54 A. Sissakian,14,a K. Sliwa,55 J. R. Smith,8 F. D. Snider,16 A. Soha,16 S. Somalwar,51 V. Sorin,4 P. Squillacioti,16
M. Stanitzki,59 R. St. Denis,20 B. Stelzer,32 O. Stelzer-Chilton,32 D. Stentz,37 J. Strologas,36 G. L. Strycker,33 Y. Sudo,54
A. Sukhanov,17 I. Suslov,14 K. Takemasa,54 Y. Takeuchi,54 J. PHYSICAL REVIEW D 82, 052002 (2010) 32Institute of Particle Physics, McGill University, Montre´al, Que´bec, Canada H3A 2T8; Simon Fraser University,
Burnaby, British Columbia, Canada V5A 1S6; University of Toronto, Toronto, Ontario, Canada M5S 1A7;
and TRIUMF, Vancouver, British Columbia, Canada V6T 2A3
33University of Michigan, Ann Arbor, Michigan 48109, USA
34Michigan State University, East Lansing, Michigan 48824, USA
35Institution for Theoretical and Experimental Physics, ITEP, Moscow 117259, Russia
36University of New Mexico, Albuquerque, New Mexico 87131, USA
37Northwestern University, Evanston, Illinois 60208, USA
38The Ohio State University, Columbus, Ohio 43210, USA
39Okayama University, Okayama 700-8530, Japan
40Osaka City University, Osaka 588, Japan
41University of Oxford, Oxford OX1 3RH, United Kingdom
42aIstituto Nazionale di Fisica Nucleare, Sezione di Padova-Trento, I-35131 Padova, Italy;
42bUniversity of Padova, I-35131 Padova, Italy
43LPNHE, Universite Pierre et Marie Curie/IN2P3-CNRS, UMR7585, Paris, F-75252 France
44University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA
45aIstituto Nazionale di Fisica Nucleare Pisa, I-56127 Pisa, Italy;
45bUniversity of Pisa, I-56127 Pisa, Italy;
45cUniversity of Siena, I-56127 Pisa, Italy;
45dScuola Normale Superiore, I-56127 Pisa, Italy
46University of Pittsburgh, Pittsburgh, Pennsylvania 15260, USA
47Purdue University, West Lafayette, Indiana 47907, USA
48University of Rochester, Rochester, New York 14627, USA
49The Rockefeller University, New York, New York 10065, USA
50aIstituto Nazionale di Fisica Nucleare, Sezione di Roma 1;
50bSapienza Universita` di Roma, I-00185 Roma, Italy
51Rutgers University, Piscataway, New Jersey 08855, USA
52Texas A&M University, College Station, Texas 77843, USA
53aIstituto Nazionale di Fisica Nucleare Trieste/Udine, I-34100 Trieste, Italy;
MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . . . (CDF Collaboration)b 1Institute of Physics, Academia Sinica, Taipei, Taiwan 11529, Republic of China
2 2Argonne National Laboratory, Argonne, Illinois 60439, USA
3 3University of Athens, 157 71 Athens, Greece
4Institut de Fisica d’Altes Energies, Universitat Autonoma de Barcelona, E-08193, Bellaterra (Barcelona), Spain
5Baylor University, Waco, Texas 76798, USA
6aIstituto Nazionale di Fisica Nucleare Bologna, I-40127 Bologna, Italy;
6bUniversity of Bologna, I-40127 Bologna, Italy
7Brandeis University, Waltham, Massachusetts 02254, USA
8University of California, Davis, Davis, California 95616, USA
9University of California, Los Angeles, Los Angeles, California 90024, USA
10Instituto de Fisica de Cantabria, CSIC-University of Cantabria, 39005 Santander, Spain
11Carnegie Mellon University, Pittsburgh, Pennsylvania 15213, USA
12Enrico Fermi Institute, University of Chicago, Chicago, Illinois 60637, USA
13Comenius University, 842 48 Bratislava, Slovakia; Institute of Experimental Physics, 040 01 Kosice, Slovakia
14Joint Institute for Nuclear Research, RU-141980 Dubna, Russia
15Duke University, Durham, North Carolina 27708, USA
16Fermi National Accelerator Laboratory, Batavia, Illinois 60510, USA
17University of Florida, Gainesville, Florida 32611, USA
18Laboratori Nazionali di Frascati, Istituto Nazionale di Fisica Nucleare, I-00044 Frascati, Italy
19University of Geneva, CH-1211 Geneva 4, Switzerland
20Glasgow University, Glasgow G12 8QQ, United Kingdom
21Harvard University, Cambridge, Massachusetts 02138, USA
22Division of High Energy Physics, Department of Physics, University of Helsinki and Helsinki Institute of Physics,
FIN-00014, Helsinki, Finland
23University of Illinois, Urbana, Illinois 61801, USA
24The Johns Hopkins University, Baltimore, Maryland 21218, USA
25Institut fu¨r Experimentelle Kernphysik, Karlsruhe Institute of Technology, D-76131 Karlsruhe, Germany
26Center for High Energy Physics, Kyungpook National University, Daegu 702-701, Korea;
Seoul National University, Seoul 151-742, Korea; Sungkyunkwan University, Suwon 440-746, Korea;
Korea Institute of Science and Technology Information, Daejeon 305-806, Korea;
Chonnam National University, Gwangju 500-757, Korea; Chonbuk National University, Jeonju 561-756, Korea
27Ernest Orlando Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA
28University of Liverpool, Liverpool L69 7ZE, United Kingdom
29University College London, London WC1E 6BT, United Kingdom
30Centro de Investigaciones Energeticas Medioambientales y Tecnologicas, E-28040 Madrid, Spain
31Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA 3University of Athens, 157 71 Athens, Greece 4Institut de Fisica d’Altes Energies, Universitat Autonoma de Barcelona, E-08193, Bellaterra (B
5 sica d’Altes Energies, Universitat Autonoma de Barcelona, E-08193, Bellaterra (Barcelona), Spain
5 Institut de Fisica d Altes Energies, Universitat Autonoma de Barcelona, E-08193, Bellaterra (Barcelona), Spain
5Baylor University, Waco, Texas 76798, USA
6 g
(
)
p
5Baylor University, Waco, Texas 76798, USA
6 5Baylor University, Waco, Texas 76798, USA 5Baylor University, Waco, Texas 76798, USA
6 Fisica Nucleare Bologna, I-40127 Bologna, Italy; uto Nazionale di Fisica Nucleare Bologna, I-40127 B
6b 6bUniversity of Bologna, I-40127 Bologna, Italy 7Brandeis University, Waltham, Massachusetts 02254, USA 8University of California, Davis, Davis, California 95616, USA 9University of California, Los Angeles, Los Angeles, California 90024, USA y
f
11Carnegie Mellon University, Pittsburgh, Pennsylvania 15213, USA g
y
g
y
12Enrico Fermi Institute, University of Chicago, Chicago, Illinois 60637, USA y
f
g
g
niversity, 842 48 Bratislava, Slovakia; Institute of Experimental Physics, 040 01 Kosice, Slovakia
14 14Joint Institute for Nuclear Research, RU-141980 Dubna, Russia or Nuclear Research, RU-141980 Dubna, Russia 15Duke University, Durham, North Carolina 27708, USA 16Fermi National Accelerator Laboratory, Batavia, Illinois 60510, USA ersity of Florida, Gainesville, Florida 32611, USA zionali di Frascati, Istituto Nazionale di Fisica Nucleare, I-00044 Frascati, Italy
19 19University of Geneva, CH-1211 Geneva 4, Switzerland
20 20Glasgow University, Glasgow G12 8QQ, United Kingdom
21 21Harvard University, Cambridge, Massachusetts 02138, USA 23University of Illinois, Urbana, Illinois 61801, USA 24The Johns Hopkins University, Baltimore, Maryland 21218, USA 28University of Liverpool, Liverpool L69 7ZE, United Kingdom
29 29University College London, London WC1E 6BT, United Kingdom o de Investigaciones Energeticas Medioambientales y Tecnologicas, E-28040 Madrid
31Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA ro de Investigaciones Energeticas Medioambientales y Tecnologicas, E-28040 Madrid,
31Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA 052002-2 PHYSICAL REVIEW D 82, 052002 (2010) PHYSICAL REVIEW D 82, 052002 (2010) and TRIUMF, Vancouver, British Columbia, Canada V6T 2A3
33 33University of Michigan, Ann Arbor, Michigan 48109, USA
4 34Michigan State University, East Lansing, Michigan 48824, USA Institution for Theoretical and Experimental Physics, ITEP, Moscow 117259, Russia
36 36University of New Mexico, Albuquerque, New Mexico 87131, USA
37 thwestern University, Evanston, Illinois 60208, USA 38The Ohio State University, Columbus, Ohio 43210, USA 39Okayama University, Okayama 700-8530, Japan y
y
p
41University of Oxford, Oxford OX1 3RH, United Kingdom 45aIstituto Nazionale di Fisica Nucleare Pisa, I-56127 Pisa, Italy;
45b 45bUniversity of Pisa, I-56127 Pisa, Italy; aDeceased
bWith visitors from bWith visitors from cUniversity of Massachusetts Amherst, Amherst, MA 01003, USA
d cUniversity of Massachusetts Amherst, Amherst, MA 01003, USA
dIstituto Nazionale di Fisica Nucleare, Sezione di Cagliari, 09042 M cUniversity of Massachusetts Amherst, Amherst, MA 01003, USA
dIstituto Nazionale di Fisica Nucleare, Sezione di Cagliari, 09042 Monserrato (Cagliari), Italy eUniversity of California Irvine, Irvine, CA 92697, USA
f fUniversity of California Santa Barbara, Santa Barbara, CA 93106, USA gUniversity of California Santa Cruz, Santa Cruz, CA 95064, USA
h hCERN,CH-1211 Geneva, Switzerland iCornell University, Ithaca, NY 14853, USA jUniversity of Cyprus, Nicosia CY-1678, Cyprus
k kUniversity College Dublin, Dublin 4, Ireland
l y
g
lUniversity of Fukui, Fukui City, Fukui Prefecture, Japan 910-0017 Universidad Iberoamericana, Mexico D.F., Mexico nIowa State University, Ames, IA 50011, USA nIowa State University, Ames, IA 50011, USA oUniversity of Iowa, Iowa City, IA 52242, USA oUniversity of Iowa, Iowa City, IA 52242, USA pKinki University, Higashi-Osaka City, Japan 577-8502 pKinki University, Higashi-Osaka City, Japan 577-8502 qKansas State University, Manhattan, KS 66506, USA qKansas State University, Manhattan, KS 66506, USA rUniversity of Manchester, Manchester M13 9PL, England rUniversity of Manchester, Manchester M13 9PL, England sQueen Mary, University of London, London, E1 4NS, England
t y, University of London, London, E1 4NS, England tMuons, Inc., Batavia, IL 60510, USA tMuons, Inc., Batavia, IL 60510, USA uNagasaki Institute of Applied Science, Nagasaki, Japan uNagasaki Institute of Applied Science, Nagasaki, Japan uNagasaki Institute of Applied Science, vNational Research Nuclear University, Moscow, Russia vNational Research Nuclear University, Moscow, Russia wUniversity of Notre Dame, Notre Dame, IN 46556, USA wUniversity of Notre Dame, Notre Dame, IN 46556, USA xUniversidad de Oviedo, E-33007 Oviedo, Spain xUniversidad de Oviedo, E-33007 Oviedo, Spain yTexas Tech University, Lubbock, TX 79609, USA yTexas Tech University, Lubbock, TX 79609, US zIFIC (CSIC-Universitat de Valencia), 56071 Valencia, Spain zIFIC (CSIC-Universitat de Valencia), 56071 Valencia, Spain aaUniversidad Tecnica Federico Santa Maria, 110v Valparaiso, Chile
bb p
bbUniversity of Virginia, Charlottesville, VA 22906, USA p
bbUniversity of Virginia, Charlottesville, VA 22906, USA ccYarmouk University, Irbid 211-63, Jordan
dd ccYarmouk University, Irbid 211-63, Jordan
dd ddOn leave from J. PHYSICAL REVIEW D 82, 052002 (2010) 53bUniversity of Trieste/Udine, I-33100 Udine, Italy
54 y
f
y
54University of Tsukuba, Tsukuba, Ibaraki 305, Japan
55Tufts University, Medford, Massachusetts 02155, USA University of Tsukuba, Tsukuba, Ibaraki 305, Japan 55Tufts University, Medford, Massachusetts 02155, USA
56 56Waseda University, Tokyo 169, Japan 57Wayne State University, Detroit, Michigan 48201, USA
58 58University of Wisconsin, Madison, Wisconsin 53706, USA
59 59Yale University, New Haven, Connecticut 06520, USA A measurement of the tt production cross section in p p collisions at
ffiffis
p ¼ 1:96 TeV using events with
two leptons, missing transverse energy, and jets is reported. The data were collected with the CDF II
detector. The result in a data sample corresponding to an integrated luminosity 2:8 fb1 is tt ¼ 6:27
0:73ðstatÞ 0:63ðsystÞ 0:39ðlumÞ pb: for an assumed top mass of 175 GeV=c2. PACS numbers: 14.65.Ha, 12.38.Qk, 13.85.Qk DOI: 10.1103/PhysRevD.82.052002 DOI: 10.1103/PhysRevD.82.052002 I. INTRODUCTION BRðWþ ! ‘þÞ BrðW ! ‘Þ 5% with ‘ ¼ e or
, but it has a signal to background ratio well above unity
even before requiring the identification of one of the jets
originating from a b quark. This analysis does not require
jets in the events to have secondary vertices consistent with
the presence of a b-hadron decay as this selection would
further reduce the acceptance by almost 50%. This paper describes a measurement of the tt production
cross section in p p collisions at
ffiffis
p ¼ 1:96 TeV with the
CDF detector at the Fermilab Tevatron. This measurement
requires the identification of both leptons in the decay
chain tt ! ðWþbÞðWbÞ ! ð‘þv‘bÞð‘ v‘bÞ. Events are
selected with two high transverse energy leptons: high
missing transverse energy (E6
T) and at least two jets in
the final state. From the excess of events selected in the
data over the predicted background from other known
standard model (SM) sources, we obtain a measurement
of the production of tt events. In Sec. II we give a short description of the detector. In
Sec. III the data sample and event selection are presented. Section IV presents the formula used for the cross section
calculation and the measurement of the tt acceptance in
dilepton events. Section V describes the calculation of the
backgrounds. Systematics uncertainties are covered in
Sec. VI. In Sec. VII observations are compared to predic-
tions in control samples characterized by the presence of
two leptons plus high E6
T in the final state. We conclude
by presenting the result of our measurement in Sec. IX,
followed by a short summary in Sec. X. The top quark pair production in the standard model
proceeds primarily by quark-antiquark annihilations. At
the Tevatron the predictions are 85% quark-antiquark
annihilations and 15% gluon fusions. At the Large
Hadron Collider at
ffiffis
p ¼ 14 TeV the situation is predicted
to be very different with 90% of the production being due
to gluon-gluon fusion and 10% due to quark-antiquark
annihilation. PHYSICAL REVIEW D 82, 052002 (2010) Stefan Institute, Ljubljana, Slovenia ddOn leave from J. Stefan Institute, Ljubljana, Slovenia 052002-3 052002-3 T. AALTONEN et al. PHYSICAL REVIEW D 82, 052002 (2010) T. AALTONEN et al. T. AALTONEN et al. III. DATA SAMPLE AND EVENT SELECTION This analysis is based on an integrated luminosity of
2:8 fb1 collected with the CDF II detector between March
2002 and April 2008. The data are collected with an
inclusive lepton trigger that requires an electron (muon)
with ET > 18 GeV (pT > 18 GeV=c). The transverse
energy and transverse momentum are defined as ET ¼
E sin and pT ¼ p sin, where E is energy measured in
the calorimeter and p is momentum measured by the
tracking system. From this inclusive lepton data set, events
with a reconstructed isolated electron of ET (muon of pT)
greater than 20 GeVðGeV=cÞ are selected. Isolation is
defined as the calorimeter energy deposited in a cone of
radius R
ffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
ðÞ2 þ ðÞ2
p
¼ 0:4 in space cen-
tered around the lepton, minus the energy deposited by the
lepton itself. Details on electron and muon identification,
or lepton ID, criteria used in this analysis are contained in
Ref. [10]. Electrons are identified by matching clusters of
localized energy deposition in the calorimeter to tracks
reconstructed using hits from the tracking chambers and
the extra constraint provided by the position of the beam
line in the transverse direction. We further require that the
energy deposition in the electromagnetic section of the
calorimeter exceeds the energy measured in the corre-
sponding hadronic section and the lateral cluster energy
profile agrees with shapes derived from electron beam test
data. Muons are identified by matching tracks to minimum
ionizing-like clusters in the calorimeter and to stubs, or sets
of radially aligned hits, in the muon chambers. Leptons
passing all of the lepton identification cuts and also having p
y
A fraction of events passing the dilepton selection does
not originate from p p collisions but from beam interac-
tions with the detector and shielding material or from
cosmic ray sources. These events are removed by requiring
a reconstructed vertex consistent with originating from the
beam interaction region and within 60 cm of the center of
the detector along the z direction. We also require that the
timing of tracks in dimuon events be consistent with both
muons traveling from the center of the detector outward
into the tracking chamber [11]. MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS isolation less than 10% of the lepton energy are defined as
‘‘tight’’. They can be of one of four categories: electrons
reconstructed in the central electromagnetic (CEM) or plug
electromagnetic (PHX from ‘‘Phoenix’’ the name of the
tracking algorithm) calorimeter and muons pointing to the
regions covered by both layers of CMU and CMP cham-
bers (CMUP) or by the central muon extension chambers
(CMX). These tight leptons are also required to be the
objects that trigger the event, with the exception of PHX
electrons which are allowed in events triggered by non-
isolated CEM, CMUP, or CMX. The calorimeter system [7] is split radially into electro-
magnetic and hadronic sections segmented in projective
tower geometry and covers the pseudorapidity range jj <
3:6. The
electromagnetic
sampling
calorimeters
are
constructed of alternating layers of lead absorber and
scintillator whereas the hadronic calorimeters use iron
absorbers. The central strip chambers are embedded in
the central electromagnetic calorimeter at a depth of about
6X0 (radiation length), which is the region of maximum
shower intensity for electrons. In the plug region stereo
layers of scintillator bars are placed at shower maximum. A set of central muon drift chambers located outside the
central calorimeters (CMU), complemented by another set
of central muon chambers set behind a 60 cm iron shield
(CMP), provides muon coverage for jj 0:6. Additional
drift chambers and scintillation counters (CMX) detect
muons in the region 0:6 jj 1:0 [8]. A second electron of ET (muon of pT) greater than
20 GeVðGeV=cÞ is also required using looser identification
cuts and without the isolation requirement. ‘‘Loose’’ lep-
tons are either electrons or muons which pass the same
identification cuts as the tight leptons but fail the isolation
requirement. Another category of loose leptons has no tight
lepton counterpart and is made of muons with tracks point-
ing to regions covered by only one of the two central muon
chambers (CMU, CMP) or with tracks of energy deposition
corresponding to a minimum ionizing particle and pointing
to regions not covered by a muon chamber (CMIO). CMIO
muons, as well as PHX electrons, must be isolated. Multicell gas Cerenkov counters [9] located in the 3:7 <
jj < 4:7 region measure the average number of inelastic
p p collisions per bunch crossing and thus determine the
beam luminosity. MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . The total uncertainty on the luminosity
is estimated to be 5.9%, of which 4.4% comes from the
acceptance and operation of the luminosity monitor and
4.0% from the uncertainty in the inelastic p p cross section. Each dilepton candidate must contain at least one tight
lepton and at most one loose lepton. These requirements
result in 18 different DIL dilepton categories, as illustrated
in Sec. IVA, where background estimates and acceptances
are calculated separately for each category. Events with
more than two tight or loose leptons in the final state are
rejected as they come mostly from background sources like
WZ and ZZ events. Another source of trilepton background
comes from Drell-Yan Z= ! eþe events with a radi-
ated photon converting into an asymmetric eþe pair, that
is a conversion pair where one electron does not reach the
minimum track pT threshold of 500 MeV=c needed for
the electron to not be trapped inside the tracking chamber. The loss in signal efficiency from removal of events with
three or more leptons is only 0.4%. II. DETECTOR This analysis improves upon a previous measurement of
the cross section using the same dilepton (DIL) selection in
a data sample with an integrated luminosity of 0:197 fb1
[1]. Unlike other CDF measurements of the tt cross section
in the dilepton channel [2], where one ‘ is identified as e or
while the other is identified by the presence of a high
momentum central track, the DIL analysis positively iden-
tifies both leptons as either electrons or muons from W
decays or as products of semileptonic decays of leptons,
thus allowing for the comparison of the observed yield of tt
decays to ee, , and e final states with the predictions
from lepton universality. CDF II is a general-purpose detector that is described in
detail elsewhere [4]. The components relevant to this
analysis are briefly described here. The detector has an
approximate full angular coverage with a charged-particle
tracker inside a magnetic solenoid, backed by calorimeters
and muon detectors. CDF uses a cylindrical coordinate
system in which is the polar angle about an axis defined
by the proton beam and is the azimuthal angle about the
beam axis. Particle pseudorapidity is defined as ¼
lntanð=2Þ. The charged-particle tracking system surrounds the
beam pipe and consists of multiple layers of silicon
micro-strip detectors, which cover a pseudorapidity region
jj < 2, and a 3.1 m long open-cell drift chamber covering
the pseudorapidity region jj < 1 [5,6]. The tracking sys-
tem is located inside a superconducting solenoid, which in
turn is surrounded by calorimeters. The magnetic field has
a strength of 1.4 Tand is aligned coaxially with the p and p
beams. The measurement provides a test of the QCD calcula-
tions of the tt cross section [3] in a channel which is
independent and complementary to other measurements
of the tt cross section in higher statistics final states where
at least one W boson from the top quark is reconstructed
via its hadronic decay, W ! qq0. The dilepton final state
suffers from a lower statistical precision, as the product
of the branching ratios of the semileptonic W decay 052002-4 PHYSICAL REVIEW D 82, 052002 (2010) PHYSICAL REVIEW D 82, 052002 (2010) PHYSICAL REVIEW D 82, 052002 (2010) The acceptance, which in our definition includes the
effects of the detector geometrical acceptance, lepton
identification, and tt to dilepton selection, is measured
using the PYTHIA Monte Carlo program [14] to simulate
tt events of all three decay modes (hadronic, lepton þ jets,
and dilepton) with an assumed Mtop ¼ 175 GeV=c2. Monte Carlo simulated events are required to have both
W bosons from top quarks decaying to a lepton plus
neutrino, where the lepton can be either an electron or
muon. The tt Monte Carlo simulation acceptance pre-
diction of 0:808 0:004ðstatÞ% is corrected by taking
into account differences observed between data and the
Monte Carlo simulation modeling of the detector response
in independent control samples. The following sections
describe the implementation and checks on the acceptance
correction procedure. to calorimeter dead zones and to nonlinear tower response
to deposited energy. These effects are convoluted to pro-
vide the jet energy scale (JES) correction factor which
estimates the energy of the originating parton from the
measured energy of a jet [13]. Jets used in the DIL selec-
tion are required to have corrected ET > 15 GeV and
jj < 2:5. jj
We further impose two cuts based on the kinematical
properties of the event: the first is a cut on the missing
transverse energy E6
T
1 which measures the transverse
energy of the neutrinos via the imbalance of the energy
detected in the calorimeter, after correcting for the pres-
ence of muons. We require that E6
T > 25 GeV, which is
strengthened to a > 50 GeV requirement if any lepton or
jet is closer than 20 to the E6
T direction. This cut, in the
following referred to as L-cut, is used to reject mainly Z ! þ and events with mismeasured E6
T generated by jets
pointing to cracks in the calorimeter. The second, or Z-veto
cut, aims at reducing the contamination of dilepton decays
of the Z boson by requiring high missing ET significance
for ee and events with dilepton invariant mass in the
76–106 GeV=c2
region. Missing ET
significance, or
MetSig, is defined as E6
T= ffiffiffiffiffiffiffiffiffiffi
Esum
T
p
, where Esum
T
is the sum
of transverse energies deposited in all calorimeter towers. 1The scalar quantity E6
T is the magnitude of the missing
transverse energy vector ~E6
T defined as the opposite of the sum
over the calorimeter towers P
iEi
Tni of the transverse energy
measured in each tower, Ei
T, times the unit vector in the azimu-
thal plane that points from the beam line to the ith calorimeter
tower, ni. PHYSICAL REVIEW D 82, 052002 (2010) This variable separates events with real E6
T due to neutrinos
from events where the E6
T is due to energy measurement
fluctuations or energy loss in calorimeter cracks. This
second category of events is expected to have a degraded
E6
T resolution. In the DIL selection, we use a cut of
MetSig > 4 GeVð1=2Þ. III. DATA SAMPLE AND EVENT SELECTION Electrons from conversions
of photons in the detector material are removed by identi-
fying events with a track near the electron track of opposite
curvature and consistent with coming from a ! eþe
vertex. Jets are reconstructed from the calorimeter towers using
a cone algorithm with fixed radius R ¼ 0:4 in
space [12]. The jet ET is corrected for detector effects due 052002-5 A. Signal acceptance The available statistics for each subsample of DIL
events can be maximized by requiring that only detector
parts essential for the identification of a particular lepton
category be fully functioning. For example, PHX electrons
require hits in the silicon inner vertex detector to recon-
struct their tracks. Hence the identification of events with
PHX electrons is limited to data taken with ‘‘good,’’ i.e. fully functioning, silicon detectors but no such require-
ments are imposed on events where the electron is central. To accommodate this approach, we rewrite the denomina-
tor of the cross section formula in Eq. (1) as the sum of the
acceptance for each DIL category Ai, weighted by the
luminosity relative to that category Li: Events in the DIL dilepton sample passing the L-cut and
Z-veto cut become tt candidate events if they have at least
2 jets, if the two leptons are of opposite charge, and if HT
the transverse energy sum of leptons, neutrinos, and jets is
greater than 200 GeV. Events in the DIL dilepton sample
reconstructed with 0 or 1 jet are used as control samples for
the background estimation. A L ¼
X
i
Ai Li:
(2) (2) In this analysis four different luminosity samples are used,
corresponding to the integrated luminosity of runs with
fully functional subdetectors for trigger CEM electrons
and CMUP muons, either ignoring the status of the
silicon detectors (2826 pb1) or requiring good silicon
(2676 pb1), and runs fully functional also for trigger
CMX muons, either ignoring (2760 pb1) or requiring
(2623 pb1) good silicon. In defining runs good for
CEM, CMUP, or CMX leptons, the distinction between
isolated and nonisolated leptons is irrelevant. IV. CROSS SECTION In this analysis we measure the cross section by using
the formula tt ¼ Nobs Nbkg
A L
;
(1) (1) The acceptances Ai can be factorized in terms of the
two leptons ‘1 and ‘2 comprising the DIL category i
according to the following formula: where Nobs is the number of dilepton candidate events, and
Nbkg is the total background. The denominator is the
product for the acceptance for tt candidate events, A,
and of the data set luminosity L. A i ¼ A‘1‘2 C‘1‘2;
(3) (3) where the A‘1‘2 are the raw PYTHIA tt Monte Carolo (MC)
efficiencies for events with reconstructed leptons ‘1 and ‘2
passing the full DIL selection and the C‘1‘2 are correction
factors specific for that lepton pair. The correction factors
are in turn calculated as 052002-6 MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . . PHYSICAL REVIEW D 82, 052002 (2010) TABLE II. List, by dilepton category, of raw acceptance
A‘1‘2, correction factor C‘1‘2, and luminosity Li used in the
calculation of the denominator for the 2:8 fb1 DIL cross section
measurement. The acceptance of each category includes contri-
butions from nonisolated loose leptons. The A‘1‘2 uncertainty
comes only from the MC statistics. The error in the C‘1‘2 comes
from the propagation of the dilepton efficiency uncertainties of
Table I. TABLE II. List, by dilepton category, of raw acceptance
A‘1‘2, correction factor C‘1‘2, and luminosity Li used in the
calculation of the denominator for the 2:8 fb1 DIL cross section
measurement. The acceptance of each category includes contri-
butions from nonisolated loose leptons. The A‘1‘2 uncertainty
comes only from the MC statistics. The error in the C‘1‘2 comes
from the propagation of the dilepton efficiency uncertainties of
Table I. where z0 is an event efficiency while trgi and SF‘i are
single lepton trigger efficiency and identification efficiency
scale factors, respectively. The factor z0 accounts for the
efficiency of 60 cm cut on the z position of the recon-
structed event vertex. By using a sample of minimally
biased inelastic interactions, we find that this cut accepts
96:63 0:04ðstatÞ% of the full CDF luminous region. The lepton trigger efficiencies trgi are measured in data
samples selected with independent sets of triggers and
found to be around 90% or better. Finally, the scale factors
SF‘i are calculated as ratios of lepton identification
efficiencies measured in data and in Monte Carlo
simulations. IV. CROSS SECTION DIL category
A‘1‘2(%)
C‘1‘2
Li(pb1)
CEM-CEM
0:1224 0:0017
0:9338 0:0019
2826
CEM-PHX
0:0470 0:0010
0:8658 0:0027
2676
CMUP-CMUP
0:0498 0:0011
0:8189 0:0025
2826
CMUP-CMU
0:0191 0:0007
0:7920 0:0040
2826
CMUP-CMP
0:0267 0:0008
0:7299 0:0035
2826
CMUP-CMX
0:0474 0:0010
0:8569 0:0020
2760
CMUP-CMIO
0:0234 0:0007
0:8050 0:0040
2826
CMX-CMX
0:0106 0:0005
0:8996 0:0027
2760
CMX-CMU
0:0075 0:0004
0:8134 0:0043
2760
CMX-CMP
0:0115 0:0005
0:7495 0:0037
2760
CMX-CMIO
0:0101 0:0005
0:8267 0:0043
2760
CEM-CMUP
0:1769 0:0020
0:8737 0:0018
2826
CEM-CMU
0:0349 0:0009
0:8879 0:0040
2826
CEM-CMP
0:0475 0:0010
0:8182 0:0035
2826
CEM-CMX
0:0845 0:0014
0:9171 0:0019
2760
CEM-CMIO
0:0410 0:0010
0:9025 0:0041
2826
PHX-CMUP
0:0327 0:0009
0:7723 0:0029
2676
PHX-CMX
0:0147 0:0006
0:7922 0:0032
2623 Table I lists all the factors used in the acceptance cor-
rection. Their central values are the luminosity weighted
averages over different data taking periods, and the quoted
uncertainties are only statistical. Using these as inputs to
Eq. (4), we obtain values ranging from 73% to 93% for the
correction factors C‘1‘2, as shown in Table II, and a total
denominator (1) for the 2:8 fb1 DIL cross section of
19:43 0:10 pb1, where the uncertainty comes solely
from the propagation of the statistical uncertainties of
each term in Eqs. (3) and (4). T. AALTONEN et al. In order to find a
consistent normalization for all data, we perform a fit to the
Z cross sections in the different dilepton categories with
three free parameters, corresponding to a multiplicative
factor in front of the selection efficiency of each of the
three loose muon categories. The fit returns the cross
sections reported in the last column of Table III and an
average Z peak cross section of 249:1 0:8 pb, as shown
in Fig. 3. The Z peak cross section measured here does not
include the 6% uncertainty in the luminosity which is
common to all channels and which is the dominant uncer-
tainty for the Z measured in [15]. The free param-
eters returned by the fit are SFZ
CMP ¼ 0:977 0:011,
SFZ
CMU ¼ 1:072 0:011, and SFZ
CMIO ¼ 0:954 0:010. They are folded into the acceptance correction procedure
as additional scale factors to be multiplied by the lepton
identification scale factors of Table I for the appropriate
categories. After the fit, the single Z cross section has been independently measured by CDF as Z ¼ 256
16 pb [15]. For the first check, we look for possible time variations
of the measured Z cross section in different data taking
periods corresponding to integrated luminosities between
200 and 500 pb1. Figure 1 shows the result of these
checks for dielectron channels. The error bar in the figure
reflects uncertainties of data statistics, Monte Carlo statis-
tics, and the acceptance correction procedure. We do not
observe any systematic trend in the time dependence of the
cross section for dielectron categories. The same conclu-
sions hold for the dimuons channels. As an example, Fig. 2
shows the Z cross section time dependence for events with
two tight muons. For the second check, Table III reports the cross sections
measured over the whole 2:8 fb1 data sample for each
dilepton category, using the efficiencies and scale factors
of Table I. The categories with two tight leptons (CEM-
CEM, CEM-PHX, CMUP-CMUP, CMUP-CMX, and CEM_CEM
CEM_PHX
CMUP_CMUP
CMUP_CMU
CMUP_CMP
CMUP_CMX
CMUP_CMIO
CMX_CMX
CMX_CMU
CMX_CMP
CMX_CMIO
[pb]
σ
200
220
240
260
280
300
-1
Z cross section for 2.8 fb
0.8 pb
±
= 249.1
σ
Fitted Z Cross section :
FIG. 3. B. Check of acceptance corrections As a cross-check of our acceptance correction proce-
dure, we calculate the cross section for Z production for
each dielectron and dimuon category used in the DIL selection. With this check we verify the consistency of
the correction procedure across the different dilepton
categories. TABLE I. Event vertex reconstruction efficiency z0 and list,
by lepton type, of trigger efficiency trg, and lepton identification
efficiency scale factors (SF) defined in Eq. (4). These are the
luminosity weighted averages of efficiencies calculated for
different data taking periods. We select events with ee or in the final state and
require the two leptons to have opposite charges and
invariant mass in the range to 76–106 GeV=c2. We follow
the same lepton pairing used for the DIL dilepton selection. Cosmic ray induced events and events with an identified
conversion are removed following the same criteria used
for the tt dilepton selection. The number of events selected
by these cuts is shown in Table III. Vertex reconstruction efficiency, z0
0:9663 0:0004
Lepton type
Trigger efficiency, trg
CEM
0:965 0:001
CMUP
0:917 0:002
CMX
0:896 0:002
Lepton identification scale factor, SF
CEM
0:987 0:001
PHX
0:932 0:002
CMUP
0:927 0:002
CMX
0:973 0:002
CMU
0:97 0:01
CMP
0:90 0:01
CMIO
0:99 0:01 We
use
Z= ! ee
and
Z= !
PYTHIA
Monte Carlo simulated samples to calculate the raw
acceptance of the selection described above in the invariant
mass 76 GeV=c2 < M‘‘ < 106 GeV=c2. We use a formu-
lation for the Z cross section calculation analogous to
the tt cross section formulation in Eq. (1). In particular
we employ the same factorization for the denominator
correction calculation prescribed in the previous section
by Eqs. (2)–(4). Lepton identification scale factor, SF We perform two checks: time independence of the
acceptance correction factor for each DIL category and
consistency of the correction procedure among different
categories. With these checks we are not trying to measure
the Z cross section but rather to determine if our under-
standing of the acceptance is correct. The Z cross section 052002-7 Dataset Periods
0d
0h
0i
P8
P9
P10
P11
P12
P13
P14
P15
P16
P17
ALL
[pb]
σ
200
220
240
260
280
300
Tight-Tight muon
→
Z
CMUP_CMUP
CMUP_CMX
CMX_CMX
FIG. 2. T. AALTONEN et al. PHYSICAL REVIEW D 82, 052002 (2010) TABLE III. Number of selected events and Z cross section in
2:8 fb1 for the different ee and dilepton categories with
dilepton invariant mass in the range 76–106 GeV=c2. Results are
given both for the original and the fitted Z cross sections. The
cross section uncertainties are only from the data statistics and
from the propagation of the uncertainty in the single lepton
efficiency of Table I. efficiency of Table I. Category
Number
of events
Z Cross section (pb)
original
Fitted
CEM-CEM
50 519
248:0 1:4
248:0 1:4
CEM-PHX
51 468
250:3 1:4
250:3 1:4
CMUP-CMUP
14 096
251:2 2:4
251:2 2:4
CMUP-CMU
5914
261:4 3:7
243:8 4:5
CMUP-CMP
6944
244:3 3:2
250:1 4:2
CMUP-CMX
20 007
247:1 2:0
247:1 2:0
CMUP-CMIO
9099
229:2 2:7
240:3 3:6
CMX-CMX
5563
250:8 3:6
250:8 3:6
CMX-CMU
3969
275:5 4:7
257:0 5:3
CMX-CMP
4280
241:8 3:9
247:6 4:8
CMX-CMIO
5771
251:0 3:6
263:2 4:4 red by CDF as Z ¼ 25
for possible time variat
ion in different data tak
grated luminosities betw
shows the result of th
. The error bar in the fig
tatistics, Monte Carlo st
ction procedure. We do
n the time dependence of
ategories. The same con
nnels. As an example, Fi
ross section (pb)
original
Fitte
248:0 1:4
248:0
250:3 1:4
250:3
251:2 2:4
251:2
261:4 3:7
243:8
244:3 3:2
250:1
247:1 2:0
247:1
229:2 2:7
240:3
250:8 3:6
250:8
275:5 4:7
257:0
241:8 3:9
247:6
251:0 3:6
263:2 Dataset Periods FIG. 2. Z cross section using dilepton categories with tight
muons as a function of data taking period. CMX-CMX), which are also the ones with the largest
acceptance, are consistent with the theoretical prediction
of 251:6þ2:8
3:1 pb [15]. Categories with a loose muon (CMP,
CMU, or CMIO) paired to a tight muon show some resid-
ual variation around the average value which is not
consistent with statistical fluctuations. B. Check of acceptance corrections Z cross section using dilepton categories with tight
muons as a function of data taking period. PHYSICAL REVIEW D 82, 052002 (2010) PHYSICAL REVIEW D 82, 052002 (2010) Dataset Periods
0d
0h
0i
P8
P9
P10
P11
P12
P13
P14
P15
P16
P17
ALL
[pb]
σ
200
220
240
260
280
300
Tight-Tight muon
→
Z
CMUP_CMUP
CMUP_CMX
CMX_CMX
PHYSICAL REVIEW D 82, 052002 (2010) T. AALTONEN et al. Z peak cross section measured for each inclusive same
flavor dilepton category using the full 2:8 fb1 data sample after
applying the loose muon scale factor. Not included is a 6%
uncertainty in the luminosity measurement common to all chan-
nels. The band represents the mean value of 249:1 0:8 pb. CEM_CEM
CEM_PHX
CMUP_CMUP
CMUP_CMU
CMUP_CMP
CMUP_CMX
CMUP_CMIO
CMX_CMX
CMX_CMU
CMX_CMP
CMX_CMIO
[pb]
σ
200
220
240
260
280
300
-1
Z cross section for 2.8 fb
0.8 pb
±
= 249.1
σ
Fitted Z Cross section : Dataset Periods
0d
0h
0i
P8
P9
P10
P11
P12
P13
P14
P15
P16
P17
ALL
[pb]
σ
200
220
240
260
280
300
-1
ee Cross Section in 2.8 fb
→
Z
CEM_CEM
PHX_CEM
FIG. 1. Z cross section using dilepton categories with two
electrons as a function of data taking period. Dataset Periods
0d
0h
0i
P8
P9
P10
P11
P12
P13
P14
P15
P16
P17
ALL
[pb]
σ
200
220
240
260
280
300
-1
ee Cross Section in 2.8 fb
→
Z
CEM_CEM
PHX_CEM FIG. 3. Z peak cross section measured for each inclusive same
flavor dilepton category using the full 2:8 fb1 data sample after
applying the loose muon scale factor. Not included is a 6%
uncertainty in the luminosity measurement common to all chan-
nels. The band represents the mean value of 249:1 0:8 pb. FIG. 3. Z peak cross section measured for each inclusive same
flavor dilepton category using the full 2:8 fb1 data sample after
applying the loose muon scale factor. Not included is a 6%
uncertainty in the luminosity measurement common to all chan-
nels. The band represents the mean value of 249:1 0:8 pb. Dataset Periods Dataset Periods FIG. 1. Z cross section using dilepton categories with two
electrons as a function of data taking period. FIG. 1. Z cross section using dilepton categories with two
electrons as a function of data taking period. 052002-8 PHYSICAL REVIEW D 82, 052002 (2010) MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . for a b or c quark to become a well-reconstructed high pT
lepton is very small. We define different categories of
fakeable leptons, one per high pT lepton category in the
DIL dilepton selection. V. BACKGROUNDS We consider four different sources of standard model
processes that can mimic the signature of dilepton plus E6
T
plus 2 or more jets signature: diboson events (WW, WZ,
ZZ, or W), Drell-Yan production of tau leptons (DY ! ), Drell-Yan production of electrons or muons with
additional E6
T (if the event is an actual Drell-Yan event,
there is no E6
T so we refer to this as fake E6
T) (DY ! ee=), and QCD production of W boson with multiple
jets in which one jet is misidentified as a lepton (‘‘W þ jet
fakes’’). The two dominant sources of background are
DY ! ee= and W þ jet fakes. These two processes
have production cross sections much larger than the tt,
but they can only contaminate the tt dilepton signature of
two leptons plus jets and large E6
T when misreconstructions
of the event create either some large fake E6
T or a jet
misidentified as a lepton. Because it is difficult to use the
Monte Carlo simulation to predict the effect of event
misreconstruction in our detector, we estimate the back-
ground from these two processes using data-based meth-
ods, as discussed in Secs. VA and V B, respectively. The
diboson and DY ! backgrounds are calculated using
Monte Carlo simulation expectation as described in
Secs. V C and V D. Corrections are applied for trigger
and lepton ID efficiencies following the same procedure
described in Sec. IVA. Jets whose full hadronic activity is limited to single
charged pions or kaons with a late shower development
or decay in flight might deposit little energy in the
calorimeter but generate hits in the muon chambers, thus
faking the signature of a muon. We define fakeable muons
as good quality tracks of pT > 20 GeV=c with E=p < 1. Depending on which muon subdetectors these tracks point
to, we label as TCMUP, TCMX, LMIO, and LMUO fake-
able muons which can fake tight CMUP, tight CMX, loose
CMIO, or loose CMU/CMP muons, respectively. Fakeable
muons that fail the isolation requirement are put together
into a single NMUO category as long as they point to any
muon subdetector. We select fakeable leptons among generic QCD jets
collected in four different control samples, whose main
trigger requirement is the presence of at least one jet of
ETrg
T
> 20, 50, 70, and 100 GeV, respectively. V. BACKGROUNDS The simpli-
fication of the jet algorithm used in these trigger selections
tends to underestimate the energy of the offline recon-
structed jets. To ensure a trigger efficiency of 90% or
greater we require the trigger jet to have reconstructed
ET greater than 35, 55, 75, and 105 GeV, respectively, in
the four jet samples. The resulting probabilities are labeled
Jet20, Jet50, Jet70, and Jet100 fake lepton probabilities. To minimize real lepton contamination, we require that
fakeables in the denominator of the fake probability fail
one or more of the standard lepton identification cuts. For
the numerator instead we require that the fakeable leptons
pass all of the lepton identification requirements. We
estimate the contamination of real leptons from W’s,
Drell-Yan, or dibosons, using Monte Carlo simulation
predictions for the number of events with one lepton and
at least one jet above the ET threshold. Our strategy for validating the background estimation is
to compare data and background estimates in the 0-jet and
1-jet bins, as discussed in Sec. VII. T. AALTONEN et al. measurements are consistent with each other within un-
certainties, with the possible exception of categories con-
taining one CMIO loose muon for which we observe a
maximum deviation equal to 10% of the average value. This systematic deviation affects only 10% of the DIL tt
raw acceptance, corresponding to the summed contribu-
tions of any dilepton pair containing a CMIO muon in
Table II. Therefore, we estimate a final 1% systematic
uncertainty on the acceptance due to the correction
procedure. p
Jets with a large fraction of neutral to charged pion
production can create signatures with low track multiplic-
ity and large energy deposition in the electromagnetic
calorimeter, thus faking the presence of electrons. We
define fakeable electrons as tracks of pT > 20 GeV=c
pointing to an electronlike cluster with energy deposition
in the electromagnetic section of the calorimeter far
exceeding the energy measured in the hadronic section,
namely, with EHAD=EEM < 0:125. Fakeable electrons are
further divided into objects that can fake CEM or PHX
electrons depending on whether their clusters belong to the
central or plug section of the calorimeter. We label them
TCEM and TPHX, respectively. Fakeable for the noniso-
lated electrons do not require isolation for the central
cluster and are called NCEM. A. W þ jet fakes 0
l
a
t
o
t :
0
0
1
t
e
j
±
0.273
NMUO
0.001
±
0.389
LMIO
0.009
±
-0.182
LMUO
0.031
±
0.047
TCMX
0.116
±
0.049
TCMUP
0.032
±
-0.109
TPHX
0.002
±
0.441
NCEM
0.001
±
-0.126
TCEM
0.004
±
0.626
1
1
0
. 0
l
a
t
o
t :
0
7
t
e
j
±
0.118
NMUO
0.001
±
0.137
LMIO
0.008
±
-0.010
LMUO
0.037
±
-0.069
TCMX
0.111
±
0.104
TCMUP
0.030
±
-0.017
TPHX
0.003
±
0.198
NCEM
0.001
±
-0.091
TCEM
0.008
±
0.375
jet20: total
0.022
±
0.076
NMUO
0.015
±
-0.181
LMIO
0.013
±
0.415
LMUO
0.377
±
-0.663
TCMX
0.308
±
-0.374
TCMUP
0.134
±
-0.102
TPHX
0.014
±
-0.147
NCEM
0.007
±
0.213
TCEM
0.036
±
-0.079
FIG. 4. Ratio of observed total number of fake leptons for each
fakeable category vs the Jet50-based prediction normalized by
the number of observed fake leptons. The predictability of the
jet50 PT dependent fake rate is good at the 30% level, as shown
by the band in the plot. When error bars are not shown they are
smaller than the dot size. closest to the energy spectrum of jets in the dilepton plus
missing ET sample. The fake probabilities for different
lepton categories show a dependence on the transverse
energy of the fakeable lepton. To properly account for
the difference in the pT spectrum of fakeable leptons in
QCD jets vs W þ jets, we calculate fake probabilities in six
pT ranges as shown in Table IV. closest to the energy spectrum of jets in the dilepton plus
missing ET sample. The fake probabilities for different
lepton categories show a dependence on the transverse
energy of the fakeable lepton. To properly account for
the difference in the pT spectrum of fakeable leptons in
QCD jets vs W þ jets, we calculate fake probabilities in six
pT ranges as shown in Table IV. The uncertainties on the fake probabilities in Table IV
are only statistical. Variations in fake probabilities between
the different QCD jet samples are used to estimate a
systematic
uncertainty in
the
lepton
fake
estimate. Figure 4 shows a comparison between the number of
fake lepton events observed in the Jet20, Jet70, and
Jet100 data sample, after integration over the full pT
spectrum, and the number predicted by the Jet50 fake
probabilities of Table IV. A. W þ jet fakes p
p
To minimize real lepton contamination, we require that
fakeables in the denominator of the fake probability fail
one or more of the standard lepton identification cuts. For
the numerator instead we require that the fakeable leptons
pass all of the lepton identification requirements. We
estimate the contamination of real leptons from W’s,
Drell-Yan, or dibosons, using Monte Carlo simulation
predictions for the number of events with one lepton and
at least one jet above the ET threshold. Events with a single W boson plus jets can simulate the
dilepton signature when one of the jets is misidentified as a
lepton. The W þ jet fake contamination is calculated in
two steps: first we extract the probability of generic QCD
jets faking the signatures of different lepton categories;
then we apply these probabilities to weight events in the
data containing one and only one high pT lepton plus jets. The fake probabilities are measured in generic jets from
QCD decays by selecting ‘‘fakeable’’ leptons, which are
jets passing minimal lepton identification criteria described
below. We do not consider separately the heavy-flavor
contribution to our backgrounds because the probability Events with a single W boson plus jets can simulate the
dilepton signature when one of the jets is misidentified as a
lepton. The W þ jet fake contamination is calculated in
two steps: first we extract the probability of generic QCD
jets faking the signatures of different lepton categories;
then we apply these probabilities to weight events in the
data containing one and only one high pT lepton plus jets. The fake probabilities are measured in generic jets from
QCD decays by selecting ‘‘fakeable’’ leptons, which are
jets passing minimal lepton identification criteria described
below. We do not consider separately the heavy-flavor
contribution to our backgrounds because the probability We use the fake lepton probability measured in the Jet50
sample as our primary estimator to apply to data events
because the jet energy spectrum in the Jet50 sample is the 052002-9 PHYSICAL REVIEW D 82, 052002 (2010) T. AALTONEN et al. (Obs-Pred)/Obs
-3
-2
-1
0
1
2
3
Using jet50 fake rates
MET>25 GeV lepton + fakeable events
8
0
0
. A. W þ jet fakes We assess a 30% systematic
uncertainty on the ability of the Jet50 fake probabilities
to predict electron and muon fake contamination in
samples with a wide range of jet energy. We define ‘‘lepton þ fakeable’’ as those events in the
central high pT lepton data sets with one and only one good
high pT lepton, E6
T > 25 GeV and a second fakeable
object failing at least one standard lepton identification
cut. The fakeable object, which can be from any of the
fakeable categories defined above, is paired to the good
lepton and treated as the second lepton in the event when
calculating any of the kinematic variables used in the top
quark DIL selection, such as dilepton invariant mass,
corrected E6
T, and HT. Jets found in a cone of R < 0:4
around the fakeable lepton are not included in the jet
multiplicity count of that event because those jets are
associated with the fake lepton in this W þ jet fake
estimation scheme. The fake lepton contamination is
calculated by weighting each ‘‘lepton þ fakeable’’ event
found in data by the fake probability in Table IV. If more
than one fakeable object is found in the event, we pair each
of them to the good lepton and add their single fake
contributions. The fake dilepton background thus calcu-
lated contains a statistical component, which is the sum of
the fake probability uncertainty itself and the statistics of
the ‘‘lepton þ fakeable’’ sample. FIG. 4. Ratio of observed total number of fake leptons for each
fakeable category vs the Jet50-based prediction normalized by
the number of observed fake leptons. The predictability of the
jet50 PT dependent fake rate is good at the 30% level, as shown
by the band in the plot. When error bars are not shown they are
smaller than the dot size. As a check, we compare the same sign ‘‘leptonþ
fakeable’’ prediction to the number of W þ jet fakes
with same sign dilepton candidates in the signal regions. We define as fake lepton charge the charge of the track TABLE IV. Jet50 fake probabilities vs fakeable lepton pT for different fakeable categories. The uncertainties are statistical only. Because of the definition of fake probability, the denominator can fluctuate to be smaller than the numerator in low statistics high pT
bins, hence fake rate values exceeding 100%. A. W þ jet fakes Jet50 fake probabilities (%) in pT range (GeV=c)
Fakeable
[20–30]
[30–40]
[40–60]
[60–100]
[100–200]
200
TCEM
4:97 0:09
3:68 0:08
2:39 0:01
2:88 0:02
3:26 0:11
4:98 3:44
NCEM
0:74 0:1
0:53 0:01
0:58 0:01
0:49 0:01
0:80 0:07
—
TPHX
12:6 0:1
13:9 0:1
12:7 0:1
20:4 0:2
26:8 0:2
65:0 57:3
TCMUP
0:99 0:04
2:30 0:10
3:82 0:14
6:30 0:28
4:13 0:47
—
TCMX
0:91 0:06
2:20 0:15
4:74 0:21
5:09 0:55
0:76 1:98
0:44 0:25
LMUO
2:48 0:05
2:88 0:14
4:41 0:23
5:45 0:47
7:51 0:64
0:41 0:26
LMIO
21:0 0:1
25:4 0:3
27:8 0:4
40:1 0:1
37:0 1:5
107:1 14:8
NMUO
0:51 0:01
0:36 0:01
0:23 0:01
0:25 0:01
0:29 0:04
0:17 0:18 052002-10 MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . . PHYSICAL REVIEW D 82, 052002 (2010) where NDT
in and NBKG
in
represent the number of events inside
the Z-window passing the L-cut in data and in non-DY MC
background predictions, respectively. Rout=in is the ratio of
Z= ! ee= events outside to inside the Z window
predicted by the ALPGEN [16] Monte Carlo generator. associated to the fakeable lepton. Same sign dilepton can-
didates are corrected for the presence of same sign pairs
coming from tt, DY, or diboson events that are simulated in
our Monte Carlo simulations. The results of this check
are shown in Table V. Although the channel shows
deviation at the 3 standard deviation level for some jet
multiplicity bins, the agreement in the final predic-
tions over all dilepton categories is at the 1 standard
deviation level. The second contribution is calculated as N high ¼ Rhigh=lowðNDT
low NBKG
low Þ;
(6) (6) where NDT
low and NBKG
low
represent the events inside the
Z-window passing the L-cut with MetSig < 4 GeVð1=2Þ
for data and for non-DY MC background predictions,
respectively. Rhigh=low is the ratio of events passing/failing
the
MetSig > 4 GeVð1=2Þ
cut
predicted
by
ALPGEN. Table VI summarizes the inputs to Eqs. (5) and (6) and
the final values of Nout and Nhigh for each jet multiplicity
bin. B. Drell-Yan to ee= background The contamination from Z= ! ee= decays is cal-
culated using a combination of data-based and MC-based
predictions. We define DIL data samples enriched in DY
events after the L-cut by inverting the Z-veto cut and
extrapolating the remaining DY contamination in the
signal region by using the relative contribution of Z= ! ee= decays passing and failing the Z-veto cut as
predicted from MC. The Z-veto cut (see Sec. III) requires
that the dilepton invariant mass be outside the Z window
region of 76–106 GeV=c2, or, if inside, that the event have
missing ET significance E6
T= ffiffiffiffiffiffiffiffiffiffi
Esum
T
p
> 4 GeVð1=2Þ. The DY contamination in the signal sample is extracted
from the Nout and Nhigh estimates in the 2 jet bin,
corrected for the efficiency of the HT > 200 GeV and of
the opposite sign lepton cuts. The combined efficiency for
these two cuts is calculated using ALPGEN simulated Z
samples and shown as OS in Table VII. We calculate the DY ! ee= contamination as the
sum of two contributions, one outside the Z window
region, Nout, and one inside the Z window with high
MetSig, Nhigh. The first contribution is calculated as The contamination of Z= ! to e events comes
mostly from cases where one of the final state muon N out ¼ Rout=inðNDT
in NBKG
in
Þ;
(5) N out ¼ Rout=inðNDT
in NBKG
in
Þ;
(5) (5) TABLE V. Comparison between the same sign dilepton fake background prediction using the
fake rate tables and the numbers of same sign dilepton candidates found in the signal region,
after MC subtraction of standard model contamination sources. Number of SS dilepton events
ee
e
All
Corrected candidates in 0 jet
13:0 4:4
11:3 3:7
14:8 5:0
39:0 7:6
Predicted candidates in 0 jet
8:1 2:6
7:8 2:7
18:0 5:6
33:9 10:4
Corrected candidates in 1 jet
7:8 3:5
0:5 1:4
24:9 5:6
33:2 6:7
Predicted candidates in 1 jet
5:1 1:6
6:9 2:2
25:4 7:8
37:3 11:4
Corrected candidates in 2 or more jets
5:0 2:7
0:7 1:0
24:7 5:2
30:4 5:9
Predicted candidates in 2 or more jets
3:8 1:2
7:5 2:4
24:5 7:6
35:9 11:0 TABLE VI. Inputs to Eqs. A. W þ jet fakes For the calculation of tt contribution to NBKG we
use the prediction of 6.7 pb for the cross section. We
later correct this iteratively to the value measured in
the data. T. AALTONEN et al. distribution function predictions to be WW ¼ 12:4
0:8 pb, WZ ¼ 3:7 0:1 pb. For the ZZ events, a cross
section ZZ ¼ 3:8 pb is assumed with an uncertainty of
20%. W decays are simulated with the BAUR Monte Carlo
generator [19]. The leading order (LO) production cross
section of W ¼ 32 3 pb is assumed and multiplied by
a K-factor of 1.36 [20] to correct for NLO effects. The W
Monte Carlo generator acceptance prediction is multiplied
by a conversion inefficiency scale factor of 1:15 0:35 to
correct for the imperfect simulation of the tracking varia-
bles used in the conversion identification algorithm. HT;OS
ee
For Nout events
0:54 0:03
0:60 0:05
For Nin events
0:95 0:01
0:99 0:01 radiates a very energetic photon. These photons, which are
almost collinear to the muon, deposit their energy in the
EM calorimeter and produce a cluster which is associated
with the original muon track and fakes the electron signa-
ture. The missed muon gives rise to a sizable E6
T in the
event, curtailing the effectiveness of the L-cut and Z-veto
to reject them. As no data-based control sample is available
for this contamination, we estimate it using Monte Carlo
simulation predictions. Monte Carlo generators do not correctly model the jet
production from hadronic radiation, as is seen by compar-
ing the jet multiplicity spectra of data and MC predictions
for ee and events in the Z peak region. Data, even
after correcting the jet multiplicity spectrum for other
SM contributions, have higher fractions of events in the 2
or more jet bins compared to predictions. We calculate jet
multiplicity scale factors CNj as ratios of data and MC
events in each jet bin, after normalizing the MC to the
number of data in the Z peak region. These scale factors,
shown in Table VIII, are used to correct the jet multiplicity
of WW and W events. A 5% systematic uncertainty on
this correction is assessed by comparing jet multiplicity
scale factors calculated with different generators. C. Diboson background The diboson processes, WW, WZ, ZZ, and W, can
mimic the signature of the tt signal via different mecha-
nisms, with real leptons and E6
T from W and Z decays and
jets produced by boson hadronic decays or initial and final
state radiation. For WW events, the two leptons and the E6
T
are produced when both W’s decay semileptonically but
the jets require some hadronic radiation external to the
diboson system. For WZ and ZZ events, the two leptons
come from the Z boson while the other W or Z boson
provides the jets via their hadronic decays. As these decays
do not contain any neutrino, some mechanism is required
to produce fake missing transverse energy. Finally for W
events, one lepton plus E6
T is generated from the semi-
leptonic W decay while the second lepton is produced from
an asymmetric conversion in which one of the two
electrons has little energy and is caught spiralling inside
the central drift chamber. Like in the WW case, the W
system is accompanied by hadronic jets. Events involving
W þ jets fake leptons, with a real lepton from the W boson
paired to a fake lepton from the hadronic decays of the
other boson, are removed from the MC to avoid double
counting. B. Drell-Yan to ee= background (5) and (6) and for each dilepton flavor and jet multiplicity. Nout and Nhigh are the final values of the
DY ! ee and background contamination outside the Z peak region and inside the Z peak region with high MetSig, respectively. TABLE VI. Inputs to Eqs. (5) and (6) and for each dilepton flavor and jet multiplicity. Nout and Nhigh are the final values of the
DY ! ee and background contamination outside the Z peak region and inside the Z peak region with high MetSig, respectively. 0-jet
1-jet
2-jets
ee
ee
ee
NDT
in
78
45
76
58
73
43
NBKG
in
29:6 1:4
20:1 1:0
16:5 1:6
11:9 1:4
17:0 1:2
16:8 1:0
Rout=in
0:39 0:05
0:45 0:09
0:31 0:05
0:31 0:06
0:32 0:02
0:26 0:02
Nout
19:1 3:7
11:3 3:2
18:2 2:8
14:2 2:5
18:2 2:8
6:8 1:7
NDT
low
65
37
69
54
65
39
NBKG
low
12:6 0:7
9:5 0:6
5:9 0:5
5:5 0:5
6:7 0:2
8:0 0:5
Rhigh=low
0:026 0:009
0:010 0:005
0:049 0:006
0:049 0:011
0:040 0:003
0:040 0:004
Nhigh
1:23 0:21
0:26 0:06
2:85 0:41
2:34 0:36
2:16 0:32
1:22 0:25 052002-11 TABLE VII. HT and opposite sign cut efficiency for the DY ! ee and background contamination in 2 jet region. The
efficiency is calculated separately for events outside the Z peak
region passing the L-cut, and for events inside the Z peak region
also passing the MetSig > 4 GeVð1=2Þ cut. HT;OS
ee
For Nout events
0:54 0:03
0:60 0:05
For Nin events
0:95 0:01
0:99 0:01
T. AALTONEN et al. PHYSICAL REVIEW D 82, 052002 (2010) T. AALTONEN et al. VI. SYSTEMATIC UNCERTAINTIES The systematic uncertainty for the cross section mea-
surement has two main contributions: systematics in the
tt dilepton acceptance and systematics in the background
estimation. We distinguish between the uncertainties
affecting only the signal or the background from the
uncertainties common to both. the fakes contamination, the 30% uncertainty on the con-
version inefficiency scale factor affecting the W contami-
nation, and the theoretical uncertainties, ranging from
2%–10%, on the production cross sections of diboson
and Z ! processes. Although large, each of these sys-
tematics affects only a fraction of the total background. Finally, a systematics common to most Monte Carlo
generator predictions of background processes with jet
production from QCD radiation comes from the 5%
uncertainty in the CNj correction factors of Sec. V C. For the signal acceptance, we consider systematic un-
certainties coming from different MC generators, different
assumed amounts of initial (ISR) and final (FSR) state
radiation in the PYTHIA Monte Carlo, calculated by com-
paring data to expectations for the pT spectrum of the
dilepton system in Drell-Yan events and for the kinematic
distributions of the underlying events and different parton
distribution functions. These sources are uncorrelated
from each other. The two remaining and largest sources
of acceptance systematics are common to signal and back-
ground Monte Carlo simulation predictions. They arise
from uncertainties in the lepton identification (ID) scale
factors and JES. Comparing the lepton ID scale factors
calculated for Z events with 0, 1, and 2 jets, we derive
a systematic uncertainty associated with the Monte Carlo
generator acceptance correction of 2%. This is added in
quadrature to the 1% systematic uncertainty on the accep-
tance correction procedure derived from measurement of
the Z cross section in different dilepton channels (see
Sec. IVA), for a total systematic uncertainty on the lepton
ID correction of 2.2%. The JES uncertainty is calculated by
measuring the shift in acceptance due to varying the jet
energy scale correction applied to each jet in the event by
1 standard deviation of its systematic uncertainty. Table IX summarizes the systematic uncertainties in the
tt acceptance separated by the contributions which are
independent and contributions which are common with
the systematic uncertainty in the background prediction. Common contributions affect both the numerator and the
denominator of Eq. (1) used to calculate the final tt cross
section. MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . . PHYSICAL REVIEW D 82, 052002 (2010) TABLE IX. Summary of systematic uncertainties affecting the
tt acceptance: The entries in the top part of the table are treated
as uncorrelated and added in quadrature when calculating their
contribution to tt cross section systematic uncertainty via the
denominator of Eq. (1); the two entries in the bottom part of the
table are also added in quadrature but their correlation with the
systematic uncertainty in the background prediction, which
appears in the numerator of Eq. (1), is taken into account
when calculating the final systematic uncertainty in the tt cross
section. The final contamination from this process is estimated
using a Monte Carlo simulation and assumes a Z !
cross section of 251:6þ2:8
3:1 pb [15]. The simulated samples
are generated using ALPGEN generator [16] that has built-in
matching of the number of jets, coupled with PYTHIA [14]
for the shower evolution, and EVTGEN [21] for the heavy-
flavor hadron decays. All simulated events were run
through the full CDF detector simulation. To correct for
NLO effects, this value is further multiplied by a K-factor
of 1.4 [22]. The MC predictions in the different jet bins are
finally rescaled by the CNj scale factors, as discussed in
Sec. V C. Source
Systematic error (%)
MC generator
1.5
ISR
1.7
FSR
1.1
PDF
0.8
Lepton ID
2.2
Jet corrections
3.2 VI. SYSTEMATIC UNCERTAINTIES Their correlation is taken into account when cal-
culating the systematic uncertainty on the measured tt
cross section. D. Drell-Yan ! background Z= ! þ decays are simulated with the ALPGEN
generator. These events can fake the dilepton plus E6
T plus
2 or more jets signature when both ’s decay semileptoni-
cally to ‘þ‘ ‘ ‘ and jets from initial and final state
radiation are present. The contamination from this process
is expected to contribute equally to the eþe and þ
categories and to be twice as big in the e
channel. The
neutrinos from the semileptonic decays tend to have
lower energy than the neutrinos in the tt dilepton sample
and align along the direction of the leptonic decay when
the Z recoils against the external jets. Hence a big fraction
of the Z= ! events are removed by the L-cut, the cut
on the event E6
T > 25 GeV or E6
T > 50 GeV in case
any lepton or jet is closer than 20 to the E6
T direction
(see Sec. III). TABLE VIII. Jet multiplicity scale factors for Z ! eþe and
Z ! þ events in the 0-jet bin (C0j), 1-jet bin (C1j), and
2-jet bin (C2j), respectively. The last column is the weighted
average of the two same flavor Z samples and it is used as the
correction factor for e reconstructed events. The uncertainties
shown here are statistical only. Only WW background contribute to the ee, , and e
final states in the same proportion as the tt signal. Diboson
processes involving a Z contribute preferentially to the
same flavor lepton categories. W events do not contrib-
ute any background to the þ category given the
negligible probability that the photon will convert to a
muon pair. Jet multiplicity scale factor
eþe
þ
‘þ‘
C0j
1:017 0:010
0:999 0:011
1:010 0:010
C1j
0:918 0:012
0:991 0:012
0:948 0:008
C2j
1:056 0:020
1:123 0:020
1:082 0:014 The WW, WZ, and ZZ processes are simulated with
the PYTHIA Monte Carlo generator. Their production cross
section is taken from the latest next-to-leading order
(NLO) MCFM (a Monte Carlo for FeMtobarn processes
at Hadron colliders) version [17] and CTEQ6 [18] parton 052002-12 VII. CONTROL SAMPLES ee þ
21:04 4:27
16:48 3:56
3:31 0:90
40:83 7:28
W þ jet fakes
12:14 3:73
14:84 4:60
67:26 20:43
94:23 26:16
Total background
57:80 8:97
48:70 7:57
110:37 21:27
216:87 32:46
tt ( ¼ 6:7 pb)
3:94 0:22
4:02 0:22
9:47 0:49
17:44 0:86
Observed
58
54
107
219 6:7þ0:7
0:9 pb [3]. The sum of the background and signal
contributions is labeled ‘‘Total SM expectation’’ and can
be compared to the number of ‘‘Observed’’ data in
2:8 fb1. Figure 8 shows the tt and the different back-
grounds overlaid on the data, for the single lepton pT,
dilepton invariant mass, the event E6
T, and HT distributions. Again there is overall good agreement between data and
total background plus tt expectations, as shown by the
probability for the
2=ndf distribution reported on the
figures. VII. CONTROL SAMPLES We use dilepton events passing both the L-cut and
Z-veto cut, but with only 0 or 1 jets as control samples
for the background calculation. The tt contamination to the
0 jet samples is negligible and the contribution to the 1 jet
sample is small. The results for the number of observed and
expected events in the various dilepton flavor categories
are shown in Table X. The tt contribution has been calcu-
lated assuming a production cross section of 6.7 pb. For the
0-jet bin data, the single largest contribution is from dibo-
son production, followed by W þ jet fakes and DY pro-
duction; for the 1-jet bin data the single largest contribution
is from W þ jet fakes, followed by diboson and DY pro-
duction. Figures 5 and 6 show the 0-jet and 1-jet data
overlaid on top of the background and tt predictions for
four different kinematic distributions: single lepton pT,
dilepton invariant mass, event missing transverse energy
E6
T, and total scalar transverse energy HT. The largest
deviation, still below the 2 standard deviation level, is in
the Njet ¼ 0 control sample for the channel. Overall
the data are in good agreement with the expectations of
background plus signal (tt). The agreement is quantified in
terms of the probability for the
2=ndf distribution and
shown as ‘‘
2 Test’’ on the figures. Uncorrelated sources of systematic uncertainties affect-
ing the backgrounds are the 30% systematic uncertainty on 052002-13 T. AALTONEN et al. PHYSICAL REVIEW D 82, 052002 (2010) T. AALTONEN et al. T. AALTONEN et al. PHYSICAL REVIEW D 82, 052002 (2010) TABLE X. Summary table, by lepton flavor content of background estimates, tt predictions, and observed events in data
corresponding to an integrated luminosity of 2:8 fb1 for the 0-jet (top) and 1-jet (bottom) bins, respectively. The quoted uncertainties
are the sum of the statistical and systematic uncertainty. TABLE X. Summary table, by lepton flavor content of background estimates, tt predictions, and observed events in data
corresponding to an integrated luminosity of 2:8 fb1 for the 0-jet (top) and 1-jet (bottom) bins, respectively. The quoted uncertainties
are the sum of the statistical and systematic uncertainty. VIII. tt USING 2 JET SELECTION As an intermediate step toward the final result, Table XI
shows the predictions for signal and background in events
with two or more jets passing all of the tt selection criteria
except the HT > 200 GeV and the opposite lepton charge
requirement. For this sample the tt signal contribution is
almost equal to the total background contribution. There is
good agreement between the predicted and observed num-
ber of events both in overall normalization and in the bin-
by-bin distribution for the same four kinematic variables
used in the 0- and 1-jet control samples, as shown in Fig. 7. The agreement is quantified in terms of the probability for
the
2=ndf distribution and shown as ‘‘
2 Test’’ on the
figures. Table XIII summarizes the background and signal pre-
dictions for the 0, 1, and 2 jet bin control samples and
for the signal sample. Figure 9 shows the overall number of
candidate events in the different jet multiplicity bins over-
laid on top of a stacked histogram of the different back-
ground components. The band gives the tt contribution
for a cross section of 6.7 pb. The hatched area represents
the uncertainty in the total background estimate. From the
difference between the observed data and the total back-
ground predictions, we measure a cross section for tt
events in the dilepton channel of VII. CONTROL SAMPLES Njet ¼ 0 control sample per dilepton flavor category
Source
ee
e
‘‘
WW
36:36 3:26
30:10 2:71
76:40 6:76
142:87 12:57
WZ
2:88 0:21
4:56 0:31
4:21 0:29
11:65 0:74
ZZ
4:13 3:19
4:25 3:28
0:41 0:32
8:79 6:78
W
14:26 4:90
0:00 0:00
13:77 2:63
28:03 7:11
DY !
0:95 0:26
0:79 0:23
2:15 0:39
3:89 0:58
DY ! ee þ
20:32 3:94
11:59 3:34
8:13 1:33
40:04 5:77
W þ jet fakes
18:72 5:73
15:29 4:93
38:47 11:72
72:49 19:02
Total background
97:64 14:47
66:58 9:23
143:54 15:65
307:76 35:84
tt ( ¼ 6:7 pb)
0:15 0:03
0:18 0:03
0:34 0:04
0:67 0:06
Observed
99
96
147
342
Njet ¼ 1 control sample per dilepton flavor category
Source
ee
e
‘‘
WW
9:74 1:08
8:73 0:97
21:55 2:31
40:01 4:24
WZ
4:95 0:25
2:66 0:15
4:11 0:21
11:72 0:52
ZZ
1:59 1:23
1:58 1:22
0:91 0:70
4:08 3:14
W
3:70 1:47
0:00 0:00
4:43 1:12
8:14 2:27
DY !
4:64 0:83
4:42 0:78
8:81 1:50
17:87 2:99
DY ! ee þ
21:04 4:27
16:48 3:56
3:31 0:90
40:83 7:28
W þ jet fakes
12:14 3:73
14:84 4:60
67:26 20:43
94:23 26:16
Total background
57:80 8:97
48:70 7:57
110:37 21:27
216:87 32:46
tt ( ¼ 6:7 pb)
3:94 0:22
4:02 0:22
9:47 0:49
17:44 0:86
Observed
58
54
107
219 Njet ¼ 1 control sample per dilepton flavor category
Source
ee
e
‘‘
WW
9:74 1:08
8:73 0:97
21:55 2:31
40:01 4:24
WZ
4:95 0:25
2:66 0:15
4:11 0:21
11:72 0:52
ZZ
1:59 1:23
1:58 1:22
0:91 0:70
4:08 3:14
W
3:70 1:47
0:00 0:00
4:43 1:12
8:14 2:27
DY !
4:64 0:83
4:42 0:78
8:81 1:50
17:87 2:99
DY ! IX. RESULTS The signal and background DIL candidate events, that is
events in the 2 or more jet samples after the final HT >
200 GeV and opposite charge requirements, are shown in
Table XII separately for the different dilepton flavor
contribution. The tt rate is computed assuming a tt pro-
duction cross section in agreement with the NLO standard
model calculation for a top mass of 175 GeV=c2, of tt ¼ 6:27 0:73ðstatÞ 0:63ðsystÞ 0:39ðlumÞ pb; tt ¼ 6:27 0:73ðstatÞ 0:63ðsystÞ 0:39ðlumÞ pb; where the first uncertainty is statistical, the second is
the convolution
of the
acceptance
and
background where the first uncertainty is statistical, the second is
the convolution
of the
acceptance
and
background 052002-14 (GeV/c)
T
Lepton P
0
20
40
60
80
100
120
140
160
180
200
Events per 10 GeV
50
100
150
200
250
300
(GeV/c)
T
Lepton P
0
20
40
60
80
100
120
140
160
180
200
Events per 10 GeV
50
100
150
200
250
300
Njet=0
)
-1
DATA (2.8 fb
(Entries : 684)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.224
2
χ
)
2
Dilepton invariant mass (GeV/c
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
20
40
60
80
100
)
2
Dilepton invariant mass (GeV/c
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
20
40
60
80
100
Njet=0
)
-1
DATA (2.8 fb
(Entries : 342)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.864
2
χ
(GeV)
T
Missing E
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
20
40
60
80
100
120
140
160
180
(GeV)
T
Missing E
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
20
40
60
80
100
120
140
160
180
Njet=0
)
-1
DATA (2.8 fb
(Entries : 342)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.063
2
χ
(GeV)
T
H
0
100
200
300
400
500
600
Events per 40 GeV
20
40
60
80
100
120
140
160
180
(GeV)
T
H
0
100
200
300
400
500
600
Events per 40 GeV
20
40
60
80
100
120
140
160
180
Njet=0
)
-1
DATA (2.8 fb
(Entries : 342)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.080
2
χ
FIG. 5. Background and top quark signal predictions overlaid on the data for 0-jet events in 2:8 fb1. MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . . MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . . . PHYSICAL REVIEW D 82, 052002 (2010) (GeV/c)
T
Lepton P
0
20
40
60
80
100
120
140
160
180
200
Events per 10 GeV
50
100
150
200
250
300
(GeV/c)
T
Lepton P
0
20
40
60
80
100
120
140
160
180
200
Events per 10 GeV
50
100
150
200
250
300
Njet=0
)
-1
DATA (2.8 fb
(Entries : 684)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.224
2
χ
MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . )
2
Dilepton invariant mass (GeV/c
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
20
40
60
80
100
)
2
Dilepton invariant mass (GeV/c
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
20
40
60
80
100
Njet=0
)
-1
DATA (2.8 fb
(Entries : 342)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.864
2
χ Events per 20 GeV
Events per 20 GeV )
2
Dilepton invariant mass (GeV/c
60
80
100
120
140
16
)
2
Dilepton invariant mass (GeV/c
60
80
100
120
140
16 (GeV/c)
T
Lepton P (GeV/c)
T
Lepton P (G
/ )
T
p
(G
/ )
T
p
(GeV)
T
Missing E
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
20
40
60
80
100
120
140
160
180
(GeV)
T
Missing E
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
20
40
60
80
100
120
140
160
180
Njet=0
)
-1
DATA (2.8 fb
(Entries : 342)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.063
2
χ (GeV)
T
H
0
100
200
300
400
500
600
Events per 40 GeV
20
40
60
80
100
120
140
160
180
(GeV)
T
H
0
100
200
300
400
500
600
Events per 40 GeV
20
40
60
80
100
120
140
160
180
Njet=0
)
-1
DATA (2.8 fb
(Entries : 342)
Syst. Uncert. MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . . = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.080
2
χ Events per 40 GeV
Events per 40 GeV FIG. 5. Background and top quark signal predictions overlaid on the data for 0-jet events in 2:8 fb1. From top left to bottom right:
two leptons transverse energy spectrum, the dilepton invariant mass, the event E6
T and HT. The hatched area represents the uncertainty
in the total background estimate. All of the results are consistent with each other. Similar
conclusions hold for the cross section of signal events
where both leptons are isolated, which is a sample exten-
sively used in other SM precision measurements like the Z
cross section measurement: All of the results are consistent with each other. Similar
conclusions hold for the cross section of signal events
where both leptons are isolated, which is a sample exten-
sively used in other SM precision measurements like the Z
cross section measurement: systematics, and the third comes from the 6% uncertainty
in the luminosity measurement. This result assumes a top
quark mass of Mt ¼ 175 GeV=c2. Studies of the DIL
selection acceptance vs Mt show an increase in acceptance
of 3% for each 1 GeV=c2, in the range 2 GeV=c2 around
the combined Tevatron top quark mass measurement of
Mt ¼ 173:1 0:6stat 1:1syst GeV=c2 [23]. The theory
cross section decreases by approximately 0.2 pb for
each 1 GeV=c2 increase over the mass range from
170–180 GeV=c2. ttðisoÞ ¼ 6:40 0:75ðstatÞ 0:49ðsystÞ pb: The luminosity error is common to all of these subsamples
and is not explicitly quoted. As a test of lepton universality, we quote the results for
the individual dilepton flavor decay modes: tt ¼ 6:27 0:73ðstatÞ 0:63ðsystÞ 0:39ðlumÞ pb; From top left to bottom right:
two leptons transverse energy spectrum, the dilepton invariant mass, the event E6
T and HT. The hatched area represents the uncertainty
in the total background estimate. MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . . . PHYSICAL REVIEW D 82, 052002 (2010) X. CONCLUSIONS ttðeeÞ ¼ 4:57 1:56ðstatÞ 0:58ðsystÞ pb
ttðÞ ¼ 7:47 1:63ðstatÞ 0:79ðsystÞ pb;
ttðeÞ ¼ 6:43 0:95ðstatÞ 0:69ðsystÞ pb: We present a measurement of the tt cross section at the
Tevatron in a sample of data corresponding to an integrated
luminosity of 2:8 fb1 collected by the CDF II detector. Using events with two leptons, large missing energy, and 052002-15 (GeV/c)
T
Lepton P
0
20
40
60
80
100
120
140
160
180
200
Events per 10 GeV
20
40
60
80
100
120
140
160
180
200
220
(GeV/c)
T
Lepton P
0
20
40
60
80
100
120
140
160
180
200
Events per 10 GeV
20
40
60
80
100
120
140
160
180
200
220
Njet=1
)
-1
DATA (2.8 fb
(Entries : 438)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.582
2
χ
)
2
Dilepton invariant mass (GeV/c
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
10
20
30
40
50
60
70
80
90
)
2
Dilepton invariant mass (GeV/c
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
10
20
30
40
50
60
70
80
90
Njet=1
)
-1
DATA (2.8 fb
(Entries : 219)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.747
2
χ
(GeV)
T
Missing E
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
20
40
60
80
100
120
140
(GeV)
T
Missing E
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
20
40
60
80
100
120
140
Njet=1
)
-1
DATA (2.8 fb
(Entries : 219)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.627
2
χ
(GeV)
T
H
0
100
200
300
400
500
600
Events per 40 GeV
10
20
30
40
50
60
70
80
90
(GeV)
T
H
0
100
200
300
400
500
600
Events per 40 GeV
10
20
30
40
50
60
70
80
90
Njet=1
)
-1
DATA (2.8 fb
(Entries : 219)
Syst. Uncert. X. CONCLUSIONS = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.627
2
χ (GeV)
T
H
0
100
200
300
400
500
600
Events per 40 GeV
10
20
30
40
50
60
70
80
90
(GeV)
T
H
0
100
200
300
400
500
600
Events per 40 GeV
10
20
30
40
50
60
70
80
90
Njet=1
)
-1
DATA (2.8 fb
(Entries : 219)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.451
2
χ Events per 40 GeV
Events per 40 GeV Events per 20 GeV
Events per 20 GeV FIG. 6. Background and top quark signal predictions overlaid on the data for 1-jet events in 2:8 fb1 From top left to bottom right:
two leptons transverse energy spectrum, the dilepton invariant mass, the event E6
T and HT. The hatched area represents the uncertainty
in the total background estimate. TABLE XI. Summary table by lepton flavor content of background estimates, tt predictions, and observed events in data
corresponding to an integrated luminosity of 2:8 fb1 for the 2 jet bin before the HT and the opposite lepton charge requirement
events. The uncertainties are the sums in quadrature of the statistical and systematic errors. The last column is the total dilepton sample
obtained as the sum of the ee, , and e contributions. TABLE XI. Summary table by lepton flavor content of background estimates, tt predictions, and observed events in data
corresponding to an integrated luminosity of 2:8 fb1 for the 2 jet bin before the HT and the opposite lepton charge requirement
events. The uncertainties are the sums in quadrature of the statistical and systematic errors. The last column is the total dilepton sample
obtained as the sum of the ee, , and e contributions. Njet 2 tt sample per dilepton flavor category
Source
ee
e
‘‘
WW
3:54 0:63
3:65 0:65
7:50 1:28
14:70 2:47
WZ
1:75 0:23
1:01 0:14
1:68 0:23
4:44 0:57
ZZ
0:83 0:65
0:74 0:58
0:47 0:37
2:04 1:59
W
0:62 0:41
0:00 0:00
1:45 0:58
2:07 0:78
DY !
2:97 0:75
3:29 0:84
6:68 1:67
12:94 3:22
DY ! X. CONCLUSIONS = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.451
2
χ
FIG. 6. Background and top quark signal predictions overlaid on the data for 1-jet events in 2:8 fb1 From top left to bottom right:
two leptons transverse energy spectrum, the dilepton invariant mass, the event E6
T and HT. The hatched area represents the uncertainty
in the total background estimate. T. AALTONEN et al. PHYSICAL REVIEW D 82, 052002 (2010) PHYSICAL REVIEW D 82, 052002 (2010) (GeV/c)
T
Lepton P
0
20
40
60
80
100
120
140
160
180
200
Events per 10 GeV
20
40
60
80
100
120
140
160
180
200
220
(GeV/c)
T
Lepton P
0
20
40
60
80
100
120
140
160
180
200
Events per 10 GeV
20
40
60
80
100
120
140
160
180
200
220
Njet=1
)
-1
DATA (2.8 fb
(Entries : 438)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.582
2
χ
T. AALTONEN et al. )
2
Dilepton invariant mass (GeV/c
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
10
20
30
40
50
60
70
80
90
)
2
Dilepton invariant mass (GeV/c
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
10
20
30
40
50
60
70
80
90
Njet=1
)
-1
DATA (2.8 fb
(Entries : 219)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.747
2
χ Events per 20 GeV
Events per 20 GeV )
2
Dilepton invariant mass (GeV/c
0
60
80
100
120
140
16
)
2
Dilepton invariant mass (GeV/c
0
60
80
100
120
140
16 (GeV/c)
T
Lepton P (GeV/c)
T
Lepton P T
(GeV)
T
Missing E
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
20
40
60
80
100
120
140
(GeV)
T
Missing E
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
20
40
60
80
100
120
140
Njet=1
)
-1
DATA (2.8 fb
(Entries : 219)
Syst. Uncert. X. CONCLUSIONS ee þ
20:33 6:00
8:04 2:73
1:76 0:72
30:13 8:54
W þ jet fakes
9:66 3:00
18:67 5:77
54:67 16:59
83:00 22:90
Total background
39:71 8:73
35:40 7:36
74:22 17:13
149:33 28:19
tt ( ¼ 6:7 pb)
31:25 1:52
32:69 1:59
74:62 3:58
138:56 6:61
Observed
58
68
143
269 052002-16 (GeV/c)
T
Lepton P
0
20
40
60
80
100
120
140
160
180
200
Events per 10 GeV
20
40
60
80
100
120
140
160
180
200
220
(GeV/c)
T
Lepton P
0
20
40
60
80
100
120
140
160
180
200
Events per 10 GeV
20
40
60
80
100
120
140
160
180
200
220
2
≥
Njet
)
-1
DATA (2.8 fb
(Entries : 538)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.940
2
χ
)
2
Dilepton invariant mass (GeV/c
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
10
20
30
40
50
60
70
80
90
)
2
Dilepton invariant mass (GeV/c
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
10
20
30
40
50
60
70
80
90
2
≥
Njet
)
-1
DATA (2.8 fb
(Entries : 269)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.334
2
χ
(GeV)
T
Missing E
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
20
40
60
80
100
(GeV)
T
Missing E
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
20
40
60
80
100
2
≥
Njet
)
-1
DATA (2.8 fb
(Entries : 269)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.194
2
χ
(GeV)
T
H
0
100
200
300
400
500
600
Events per 40 GeV
10
20
30
40
50
60
70
80
90
(GeV)
T
H
0
100
200
300
400
500
600
Events per 40 GeV
10
20
30
40
50
60
70
80
90
2
≥
Njet
)
-1
DATA (2.8 fb
(Entries : 269)
Syst. Uncert. MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . . . = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.190
2
χ Events per 20 GeV
Events per 20 GeV (GeV)
T
Missing E (GeV)
T
Missing E FIG. 7. Background and top quark signal predictions overlaid on the data for 2-jet events before the HT and the opposite lepton
charge requirement in 2:8 fb1. From top left to bottom right: Two leptons transverse energy spectrum, the dilepton invariant mass, the
event E6
T and HT. The hatched area represents the uncertainty in the total background estimate. TABLE XII. Summary table by lepton flavor content of background estimates, tt predictions, and observed events in the final sample
of events with 2 jets passing all candidate selection criteria, for an integrated luminosity of 2:8 fb1. The uncertainties are the sums
in quadrature of the statistical and systematic errors. The last column is the total dilepton sample obtained as the sum of the ee, ,
and e contributions. TABLE XII. Summary table by lepton flavor content of background estimates, tt predictions, and observed events in the final sample
of events with 2 jets passing all candidate selection criteria, for an integrated luminosity of 2:8 fb1. The uncertainties are the sums
in quadrature of the statistical and systematic errors. The last column is the total dilepton sample obtained as the sum of the ee, ,
and e contributions TABLE XII. Summary table by lepton flavor content of background estimates, tt predictions, and observed events in the final sample
of events with 2 jets passing all candidate selection criteria, for an integrated luminosity of 2:8 fb1. The uncertainties are the sums
in quadrature of the statistical and systematic errors. The last column is the total dilepton sample obtained as the sum of the ee, ,
and e contributions. tt signal events per dilepton flavor category
Source
ee
e
‘‘
WW
2:16 0:38
2:42 0:42
4:79 0:80
9:37 1:51
WZ
0:94 0:15
0:68 0:11
0:59 0:10
2:22 0:33
ZZ
0:65 0:51
0:64 0:50
0:23 0:18
1:51 1:18
W
0:23 0:25
0:00 0:00
0:00 0:00
0:23 0:25
DY !
1:67 0:32
1:76 0:34
3:87 0:72
7:29 1:34
DY ! MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . . . PHYSICAL REVIEW D 82, 052002 (2010) MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . . . (GeV/c)
T
Lepton P
0
20
40
60
80
100
120
140
160
180
200
Events per 10 GeV
20
40
60
80
100
120
140
160
180
200
220
(GeV/c)
T
Lepton P
0
20
40
60
80
100
120
140
160
180
200
Events per 10 GeV
20
40
60
80
100
120
140
160
180
200
220
2
≥
Njet
)
-1
DATA (2.8 fb
(Entries : 538)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.940
2
χ
MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . )
2
Dilepton invariant mass (GeV/c
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
10
20
30
40
50
60
70
80
90
)
2
Dilepton invariant mass (GeV/c
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
10
20
30
40
50
60
70
80
90
2
≥
Njet
)
-1
DATA (2.8 fb
(Entries : 269)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.334
2
χ Events per 20 GeV
2
3
4
5
6
7
8
Events per 20 GeV
2
3
4
5
6
7
8 )
2
Dilepton invariant mass (GeV/c )
2
Dilepton invariant mass (GeV/c (GeV/c)
T
Lepton P (GeV/c)
T
Lepton P (GeV)
T
Missing E
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
20
40
60
80
100
(GeV)
T
Missing E
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
20
40
60
80
100
2
≥
Njet
)
-1
DATA (2.8 fb
(Entries : 269)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.194
2
χ (GeV)
T
H
0
100
200
300
400
500
600
Events per 40 GeV
10
20
30
40
50
60
70
80
90
(GeV)
T
H
0
100
200
300
400
500
600
Events per 40 GeV
10
20
30
40
50
60
70
80
90
2
≥
Njet
)
-1
DATA (2.8 fb
(Entries : 269)
Syst. Uncert. X. CONCLUSIONS = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.190
2
χ
FIG. 7. Background and top quark signal predictions overlaid on the data for 2-jet events before the HT and the opposite lepton
charge requirement in 2:8 fb1. From top left to bottom right: Two leptons transverse energy spectrum, the dilepton invariant mass, the
event E6
T and HT. The hatched area represents the uncertainty in the total background estimate. MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . . . PHYSICAL REVIEW D 82, 052002 (2010) MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . . . ee þ
11:81 2:16
5:32 1:23
1:36 0:60
18:49 2:73
W þ jet fakes
3:91 1:28
9:34 3:05
20:90 6:43
34:15 9:51
Total background
21:37 3:14
20:16 3:64
31:73 6:78
73:26 11:30
tt ( ¼ 6:7 pb)
28:80 1:41
31:24 1:52
70:15 3:36
130:19 6:21
Total SM expectation
50:17 4:25
51:40 5:00
101:88 10:06
203:45 17:33
Observed
41
55
99
195 052002-17 (GeV/c)
T
Lepton P
0
20
40
60
80
100
120
140
160
180
200
Events per 10 GeV
20
40
60
80
100
120
(GeV/c)
T
Lepton P
0
20
40
60
80
100
120
140
160
180
200
Events per 10 GeV
20
40
60
80
100
120
Candidates
)
-1
DATA (2.8 fb
(Entries : 390)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.981
2
χ
)
2
Dilepton invariant mass (GeV/c
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
10
20
30
40
50
60
)
2
Dilepton invariant mass (GeV/c
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
10
20
30
40
50
60
Candidates
)
-1
DATA (2.8 fb
(Entries : 195)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.134
2
χ
(GeV)
T
Missing E
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
10
20
30
40
50
60
70
80
90
(GeV)
T
Missing E
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
10
20
30
40
50
60
70
80
90
Candidates
)
-1
DATA (2.8 fb
(Entries : 195)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.370
2
χ
(GeV)
T
H
0
100
200
300
400
500
600
Events per 40 GeV
10
20
30
40
50
60
70
80
(GeV)
T
H
0
100
200
300
400
500
600
Events per 40 GeV
10
20
30
40
50
60
70
80
Candidates
)
-1
DATA (2.8 fb
(Entries : 195)
Syst. Uncert. MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . . . = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.096
2
χ (GeV)
T
Missing E
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
10
20
30
40
50
60
70
80
90
(GeV)
T
Missing E
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
10
20
30
40
50
60
70
80
90
Candidates
)
-1
DATA (2.8 fb
(Entries : 195)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.370
2
χ Events per 40 GeV
Events per 40 GeV Events per 20 GeV
Events per 20 GeV FIG. 8. Background and top quark signal predictions overlaid on the data for top quark DIL candidate events in 2:8 fb1. From top
left to bottom right: Two leptons transverse energy spectrum, the dilepton invariant mass, the event E6
T and HT. The hatched area
represents the uncertainty in the total background estimate. TABLE XIII. Summary table by jet multiplicity bin of background estimates, tt predictions, and observed events in data
corresponding to an integrated luminosity of 2:8 fb1. The uncertainties are the sums in quadrature of the statistical and systematic
error. The last column contains the candidate events with HT > 200 GeV and opposite sign lepton cuts applied. TABLE XIII. Summary table by jet multiplicity bin of background estimates, tt predictions, and observed events in data
corresponding to an integrated luminosity of 2:8 fb1. The uncertainties are the sums in quadrature of the statistical and systematic
error. The last column contains the candidate events with HT > 200 GeV and opposite sign lepton cuts applied. corresponding to an integrated luminosity of 2:8 fb
. The uncertainties are the sums in quadrature of the statistical and systematic
error. The last column contains the candidate events with HT > 200 GeV and opposite sign lepton cuts applied. MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . . . = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.096
2
χ
FIG. 8. Background and top quark signal predictions overlaid on the data for top quark DIL candidate events in 2:8 fb1. From top
left to bottom right: Two leptons transverse energy spectrum, the dilepton invariant mass, the event E6
T and HT. The hatched area
represents the uncertainty in the total background estimate. T. AALTONEN et al. PHYSICAL REVIEW D 82, 052002 (2010) PHYSICAL REVIEW D 82, 052002 (2010) T. AALTONEN et al. (GeV/c)
T
Lepton P
0
20
40
60
80
100
120
140
160
180
200
Events per 10 GeV
20
40
60
80
100
120
(GeV/c)
T
Lepton P
0
20
40
60
80
100
120
140
160
180
200
Events per 10 GeV
20
40
60
80
100
120
Candidates
)
-1
DATA (2.8 fb
(Entries : 390)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.981
2
χ
T. AALTONEN et al. )
2
Dilepton invariant mass (GeV/c
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
10
20
30
40
50
60
)
2
Dilepton invariant mass (GeV/c
0
20
40
60
80
100
120
140
160
180
200
Events per 20 GeV
10
20
30
40
50
60
Candidates
)
-1
DATA (2.8 fb
(Entries : 195)
Syst. Uncert. = 6.7 pb)
σ
t (
t
Diboson
µ
µ
ee+
→
DY
ττ
→
DY
Fakes
Test : 0.134
2
χ Events per 20 GeV
Events per 20 GeV Events per 10 GeV
Events per 10 GeV )
2
Dilepton invariant mass (GeV/c )
2
Dilepton invariant mass (GeV/c (GeV/c)
T
Lepton P (GeV/c)
T
Lepton P (GeV)
T
H
0
100
200
300
400
500
600
Events per 40 GeV
10
20
30
40
50
60
70
80
(GeV)
T
H
0
100
200
300
400
500
600
Events per 40 GeV
10
20
30
40
50
60
70
80
Candidates
)
-1
DATA (2.8 fb
(Entries : 195)
Syst. Uncert. ACKNOWLEDGMENTS We thank the Fermilab staff and the technical staffs of the
participating institutions for their vital contributions. This
work was supported by the U.S. Department of Energy and
National Science Foundation ; the Italian Istituto Nazionale
di Fisica Nucleare; the Ministry of Education, Culture,
Sports, Science and Technology of Japan; the Natural
Sciences and Engineering Research Council of Canada;
the National Science Council of the Republic of China;
the Swiss National Science Foundation; the A. P. Sloan
Foundation; the Bundesministerium fu¨r Bildung und
Forschung,
Germany;
the
World
Class
University
Program, the National Research Foundation of Korea; the
Science and Technology Facilities Council and the Royal
Society, UK; the Institut National de Physique Nucleaire et
Physique des Particules/CNRS; the Russian Foundation for
Basic Research; the Ministerio de Ciencia e Innovacio´n,
and Programa Consolider-Ingenio 2010, Spain; the Slovak
R&D Agency; and the Academy of Finland. HT>200+OS
HT>200+OS FIG. 9. Dilepton candidate events (black point) by jet multi-
plicity. The stacked histogram represents the background con-
tribution and the tt contribution for an assumed tt ¼ 6:7 pb. The hatched area is the uncertainty in the total background
estimate. two or more jets we select a sample with a signal top
quark pair almost a factor of 2 larger than the background. The contamination from SM sources is checked in lower
jet multiplicity samples. From the excess of data over the
background predictions we measure: [1] D. Acosta et al. (CDF Collaboration), Phys. Rev. Lett. 93,
142001 (2004). [9] D. Acosta et al., Nucl. Instrum. Methods Phys. Res., Sect. A 494, 57 (2002). [2] T. Aaltonen et al. (CDF Collaboration), Phys. Rev. D 79,
112007 (2009). [10] A. Abulencia et al. (CDF Collaboration), J. Phys. G 34,
2457 (2007). [11] A. Affolder et al. (CDF Collaboration), Phys. Rev. Lett. 88, 041801 (2002). [3] M. Cacciari et al., J. High Energy Phys. 09 (2008) 127; N. Kidonakis and R. Vogt, Phys. Rev. D 78, 074005 (2008);
S. Moch and P. Uwer, Nucl. Phys. B, Proc. Suppl. 183, 75
(2008). [12] F. Abe et al. (CDF Collaboration), Phys. Rev. D 45, 1448
(1992). [4] R. Blair et al., The CDF II Detector Technical Design
Report, Fermilab-Pub-96/390-E. [13] A. Bhatti et al., Nucl. Instrum. Methods Phys. Res., Sect. A 566, 375 (2006). [14] T. Sjostrand, S. Mrenna, and P. Skands, J. High Energy
Phys. 05 (2006) 026. We use version 6.216. [5] C. S. tt ¼ 6:27 1:03ðtotalÞ pb; tt ¼ 6:27 1:03ðtotalÞ pb; consistent with the NLO standard model calculation
of 6:7þ0:7
0:9 pb. Yields in the ee, , and e final states
are in agreement with the predictions from lepton
universality. MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . . . Control sample and signal events per jet multiplicity
Source
0 jet
1 jet
2 jet
HT þ OS
WW
142:87 12:57
40:01 4:24
14:70 2:47
9:37 1:51
WZ
11:65 0:74
11:72 0:52
4:44 0:57
2:22 0:33
ZZ
8:79 6:78
4:08 3:14
2:04 1:59
1:51 1:18
W
28:03 7:11
8:14 2:27
2:07 0:78
0:23 0:25
DY !
3:89 0:58
17:87 2:99
12:94 3:22
7:29 1:34
DY ! ee þ
40:04 5:77
40:83 7:28
30:13 8:54
18:49 2:73
W þ jet fakes
72:49 19:02
94:23 26:16
83:00 22:90
34:15 9:51
Total background
307:76 35:84
216:87 32:46
149:33 28:19
73:26 11:30
tt ( ¼ 6:7 pb)
0:67 0:06
17:44 0:86
138:56 6:61
130:19 6:21
Total SM expectation
308:43 35:87
234:31 33:28
287:89 34:70
203:45 17:33
Observed
342
219
269
195 052002-18 0 jet
1 jet
2 jet
≥
HT>200+OS
Events
0
100
200
300
400
500
0 jet
1 jet
2 jet
≥
HT>200+OS
Events
0
100
200
300
400
500
data
uncertainty
σ
1
±
Bkgd
= 6.7 pb)
σ
t (
t
WW/WZ/ZZ
DY
fake
FIG. 9. Dilepton candidate events (black point) by jet multi-
plicity. The stacked histogram represents the background con-
tribution and the tt contribution for an assumed tt ¼ 6:7 pb. The hatched area is the uncertainty in the total background
estimate. MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS . PHYSICAL REVIEW D 82, 052002 (2010) 0 jet
1 jet
2 jet
≥
HT>200+OS
Events
0
100
200
300
400
500
0 jet
1 jet
2 jet
≥
HT>200+OS
Events
0
100
200
300
400
500
data
uncertainty
σ
1
±
Bkgd
= 6.7 pb)
σ
t (
t
WW/WZ/ZZ
DY
fake
MEASUREMENT OF THE TOP PAIR PRODUCTION CROSS tt ¼ 6:27 0:73ðstatÞ 0:63ðsystÞ 0:39ðlumÞ pb; tt ¼ 6:27 0:73ðstatÞ 0:63ðsystÞ 0:39ðlumÞ pb; or [20] U. Baur, T. Han, and J. Ohnemus, Phys. Rev. D 48, 5140
(1993).
[21] D. Lange, Nucl. Instrum. Methods Phys. Res., Sect. A 462,
152 (2001).
[22] T. Aaltonen et al. (CDF Collaboration), Phys. Lett. B 692,
232 (2010).
[23] Tevatron ElectroWeak Working Group for the CDF and
D0 Collaborations, arXiv:0903.2503.
T. AALTONEN et al.
PHYSICAL REVIEW D 82, 052002 (2010) [20] U. Baur, T. Han, and J. Ohnemus, Phys. Rev. D 48, 5140
(1993).
[21] D. Lange, Nucl. Instrum. Methods Phys. Res., Sect. A 462,
152 (2001).
[22] T. Aaltonen et al. (CDF Collaboration), Phys. Lett. B 692,
232 (2010).
[23] Tevatron ElectroWeak Working Group for the CDF and
D0 Collaborations, arXiv:0903.2503. [21] D. Lange, Nucl. Instrum. Methods Phys. Res., Sect. A 462,
152 (2001).
[23] Tevatron ElectroWeak Working Group for the CDF and
D0 Collaborations, arXiv:0903.2503. T. AALTONEN et al.
PHYSICAL REVIEW D 82, 052002 (2010) ACKNOWLEDGMENTS Hill et al., Nucl. Instrum. Methods Phys. Res., Sect. A 530, 1 (2004); A. Sill et al., Nucl. Instrum. Methods
Phys. Res., Sect. A 447, 1 (2000); A. Affolder et al., Nucl. Instrum. Methods Phys. Res., Sect. A 453, 84 (2000). [15] A. D. Martin, W. J. Stirling, R. S. Thorne, and G. Watt
(CDF Collaboration), Phys. Lett. B 652, 292 (2007). [6] A. Affolder et al., Nucl. Instrum. Methods Phys. Res.,
Sect. A 526, 249 (2004). [16] M. L. Mangano et al., J. High Energy Phys. 07 (2003) 001. We use version 2.10. M. L. Mangano, M. Moretti, F. Piccinini, and M. Treccani, J. High Energy Phys. 01
(2007) 013. [7] L. Balka et al., Nucl. Instrum. Methods Phys. Res., Sect. A
267,272(1988);S.Bertoluccietal.,Nucl.Instrum.Methods
Phys. Res., Sect. A 267, 301 (1988); M. Albrow et al., Nucl. Instrum. Methods Phys. Res., Sect. A 480, 524 (2002). [17] J. M. Campbell and R. K. Ellis, Phys. Rev. D 60, 113006
(1999). [18] W. Giele et al., arXiv:hep-ph/0204316. [8] G. Ascoli et al., Nucl. Instrum. Methods Phys. Res., Sect. A 268, 33 (1988). [19] U. Baur and E. L. Berger, Phys. Rev. D 41, 1476 (1990). 052002-19 T. AALTONEN et al. 052002-20
|
https://openalex.org/W4229848643
|
https://www.qeios.com/read/I73NOB/pdf
|
English
| null |
Laryngeal Obstruction, CTCAE
|
Definitions
| 2,020
|
cc-by
| 65
|
Qeios · Definition, February 2, 2020 Open Peer Review on Qeios Open Peer Review on Qeios Laryngeal Obstruction, CTCAE National Cancer Institute National Cancer Institute Qeios ID: I73NOB · https://doi.org/10.32388/I73NOB National Cancer Institute. Laryngeal Obstruction, CTCAE. NCI Thesaurus. Code C143639. National Cancer Institute. Laryngeal Obstruction, CTCAE. NCI Thesaurus. Code C143639. A disorder characterized by blockage of the laryngeal airway. Qeios ID: I73NOB · https://doi.org/10.32388/I73NOB 1/1
|
https://openalex.org/W4220727907
|
https://zenodo.org/records/6344859/files/39.pdf
|
English
| null |
Mongol Migration To Egypt And Bilad Al-Sham In Mamluk Era For The Period (658 - 694 AH/ 1260 - 1296 AD)
|
Zenodo (CERN European Organization for Nuclear Research)
| 2,022
|
cc-by
| 2,662
|
Multicultural Education Multicultural Education Volume 8, Issue 1, 2022 Mongol Migration To Egypt And Bilad Al-Sham In Mamluk Era For The
Period (658 - 694 AH/ 1260 - 1296 AD)
Abdul Khaliq Khames Ali, Zainab Adnan Hashem
Article Info
Abstract
Article History
Received:
June 28, 2021
The study addresses Mongol migration during the period ofAL-Sultan AL-
ZaherBibars reign (658-676AH/ 1260-1277 AD)beside the sultan's
perception and the his procedure's in dealing with Mongols, the search also
presents the most significant migration during AL-Sultan Qalawn reign and
his son AL-khalil(678-693AH/1279-1293AD) in addition the reasons behind
the migration decreasing as compared with AL-ZaherBibars reign. Accepted:
January 29, 2022 Mongol Migration To Egypt And Bilad Al-Sham In Mamluk Era For The
Period (658 - 694 AH/ 1260 - 1296 AD)
A
i
A i
i
A The study addresses Mongol migration during the period ofAL-Sultan AL-
ZaherBibars reign (658-676AH/ 1260-1277 AD)beside the sultan's
perception and the his procedure's in dealing with Mongols, the search also
presents the most significant migration during AL-Sultan Qalawn reign and
his son AL-khalil(678-693AH/1279-1293AD) in addition the reasons behind
the migration decreasing as compared with AL-ZaherBibars reign. Introduction Introduction
The topic of Mongol migration to Egypt definatly in the state of navel Mamluk is considered as one of the most
difficult and cross_ cutting , the migrations had an effective role on the state and Mumluk community specially
in politics , military and society During the period (658-696AH/1260-1297AD) , an important migrations had
took place specially during the reign of Al-Sultan AL-ZaherBibars and Al-sultan kitbugha and AL-Sultan
received them and welcomed AL-Mogolwormly as aresult, Mogol's migrations increased a matter rose Al-
Sultan's fears and concerns of Mogol . Al-Sultan AL-ZaherBibars thought of Mogol increasing as a plan set up
by his enemies, there for, he put soliders on alarm. The research is divided into introduction,Two pants and conclusions. First part deals with Mogol migrations
during the reign Al-Sultan AL-ZaherBibars and second part deals with Mogol's migration during the reign of
Al-Sultan Qalawn and Al-Khalil (his son) reign finally the third part concerns with the migrations during the
reign of Al-Sultan Kitbugha. Firstly: Mongol's migration during the reign of AL-Sultan AL-ZaherBibars (658-676 AH/1260-1277 AD(:
During Al-Sultan Bibars reign the first migrations has flocked to Egypt , many migrations ( individual or
group's) had been witnessed and Al-Sultan of Momluk seemed to relay on Mongol in his army's formation, It
can be argued that some factors increased migrations such as the good relationships among Mamluk , golden
trips Mongol and Persian Mongol's which contributed in flocking Mongol in addition to the bad relationship
between persion Mongol's and their judges , some person Mongol's escaped and sattled down in Egypt since it
represented as a safe haven for their families represented as a safe haven for their families IbnKatheer and AL-Maqrizi asserted the fact that , the initial Mongol migrations took place during the reign of
AL-Sultan AL-ZaherBibars in 660AH/1262AD , IbnKatheer states that Tatar delegations arrived to Al-Sultan in
hope find safety and Al-Sultan , by his role, honoredthem and was very generous supplying them efficient
incomes , In the vein , AL-Maqrizi mentions these events by details and exposes the migrations , The king
Baraka sent Mongol to stand with Holaqo but he ordered them Holaqo and Baraka Khan , had broken down and
to informed to join the Egyptian military if they couldn't manage to join him. Introduction AL-Sultan supported them and hosted them well in Bilad AL-Sham and later to Cairo there Al-Sultan by
himself recieved them in( 26 Thi AL-Hijja 660AH/21 November 1262 AD) and built them houses mating all
their needs , horses , money . they almost were two hundred knights with their families, they got a good living
and became muslims. AL-Sultan supported them and hosted them well in Bilad AL-Sham and later to Cairo there Al-Sultan by
himself recieved them in( 26 Thi AL-Hijja 660AH/21 November 1262 AD) and built them houses mating all
their needs , horses , money . they almost were two hundred knights with their families, they got a good living
and became muslims. The Sultan's good reception and generosity attracted the other Mongol migrants . AL-Maqrizi indicated that
Mongol groups flocked as a result of the sultan's generosity and good hospitality that led to increase Mongol's
migrant numbers to Egyp obviously , the text above shows the main reasons behind Mongol influx which is due
good hospitality and generosity consequently , many sects and Mongol groups were headed to Egypt , besides
thatIt seemed that the sultan's desire to put migrants in his side and his military since them had great knowledge
in military potential, moreover he wanted them to join Islam. 322 323 AL-Maqrizi mentioned that an arrival of new group in 661AH/ 1263AD who escaped from Al-Ilkhauian
according to the Sultan Baraka Khan's order , Tattar arrived with their notables Qarmon , Amtakhia , Narkia ,
Jabric , Qian , Nasisa , Teshor , Nabto , Subhi , Jarjilan , Ajqarqa , Mutaqadim , The sultan rode his hours to
meet them , as soon they saw him , They went down and kissed the ground under his feet , and he empored them
, AL-Maqrizi states that , when The Sultan AL-ZaherBibars knew of migrants coming he celebrated and
welcomed Them and did the same thing with the coming groups , He called them upon Islam and they accepted
happily,Migration to Egypt continued for the above mentioned reasonsAL-Maqrizi indicated this events stating
that influx came from Shiraz on the top of them , The prince Saif AL-DeenBiklik (i) and much princes of
Khafaja
(
ii
) they were received kindly by the sultan . Introduction j
y
y y
As the number of Mongol migrant raised the sultan got suspicious and afraid of these frequent migration ,
he viewed them as a part of policy set up by enemies , AL-Maqrizi said that many Barbarians and thieves
arrived to Egypt so the sultan gathered princes to express his fears against these continuous migrations , then he
came up with a decision , if Tattar obeyed rules then they would treated well and rewarded gift, horses and
camels and money. y
Al-Sultan AL-ZaherBibars crowed his son as a king to run to state affairs in his father absence. Al S l
d hi
i
i h
i
h
li
d i AL L
h li i
f C i y
Al-Sultan AL-ZaherBibars crowed his son as a king to run to state affairs in his father absence. Al-Sultan never stopped his generosity with migrants , they lived in AL-Luoq on the limits of Cairo Al-Sultan AL-ZaherBibars crowed his son as a king to run to state affairs in his father absence. Al-Sultan never stopped his generosity with migrants , they lived in AL-Luoq on the limits of Cairo
AL-Qalqashandi showed his hatred of Mongol saying that AL-Luoq was well built but badly inhabited by
Barbarian Al-Sultan never stopped his generosity with migrants , they lived in AL-Luoq on the limits of Cairo
AL-Qalqashandi showed his hatred of Mongol saying that AL-Luoq was well built but badly inhabited by
Barbarian L-Qalqashandi showed his hatred of Mongol saying that AL-Luoq was well built but badly inhabite
arbarian On 17thShawal 662 AH/12th of October 1264 AD , The Sultan informed of arrival another Mongol's trips so
he ordered princes of Khafaja to behave generously with them . As mentioned before , outlaws found Egypt as a safe haven not only civil people emigrated but significant
figures in AL-Elikhanian state escaped from their judges like the prince Shams AL-DeenBahadir
Three thousands migrants approached to Egypt during the reign of the Sultan Bibars , some of them were
Khazakia (knights be longed to the sultan) and AL Bira Thus , the migrants of Mongol continued throughout the reign of the Sultan Bibars , IbnShadad said that the
preparation of the Mongol coming to Egypt and Bilad AL-Sham daring his era of about three thousands same . Finally :
h d b Finally :
It had been argued that the period of The Sultan Bibars was completely full of Mongol migrants to Egypt and
Bilad AL-Sham , some sources asserts that three thousands migrants arrived to Egypt and this due to the good
hospitality, on the other hand the period (678_693AH/1274_1293AD) had witnessed a great decreasing of
migration and the reason of that due to the important of Bilad AL-Sham condition during the reign of Ahmed
Takodar beside the truce with Mongol . gg
Secondly : Migration of Mongol during the rule of the Sultan AL-Mansur Qalawon and his son AL-
Ashraf Khalil (678-693AH /1279-1293 AD) : During the reign of The Sultan Al-Mansur Qalawon , a significant decrease in migration was witnessed as
compared with AL-ZaherBirbas reign, never the less , migration never stopped at that time , sincemany unrests
had been accursed in the state period of Ahmed Takwdar reign and his nephew attempt to change the
government , all these factors effected badly on the economic of the country a matter forced Mongol to run
away. Regarding the period of the Sultan AL-Ashraf Khalil reign , Mongol migration decreased , only two
hundred migrant arrived to Egypt, The sultan recieved them and empowered them, the reasons of immigration
still ambiguous , yet . It may be due to the judge's in justice mismanagement besides an economic factors that had effected badly
on people.. Introduction (
AL-Maqrizi said that saying the magnifying of the Mongol in the reign of the Sultan AL-ZaherBibars , there
was copied Egypt and Bilad AL-Sham . It's notable that the increasing of Mongol numbers let to overcrowding in AL-Luoq area in Cairo which
considered as their in habitat , Therefore sultan ordered to build them houses inside castlethat are known as
ruins of Tattars
(
iii
) , It's essential to note that the Sultan AL-ZaherBibars was very keen to isolate the groups in
special places , and this due to the fact that AL-Mamalik always felt alienated in the state of Egyptian
and Bilad AL-Sham , They were conscious of the differences between them , there for they lived as a stranger
(
iv
). Maybe you come to mind some of the questions , Why did The sultan choose AL-Luoq in specific ? The
answer is related to the geographic position of this city. The sultan needed militant groups in the coastal area and Al 1ng was the best to pace any external
aggression gg
Secondly : Migration of Mongol during the rule of the Sultan AL-Mansur Qalawon and his son AL-
Ashraf Khalil (678-693AH /1279-1293 AD) : References Nawar, Communities of Mongol , p27 . 324 IbnKatheer , Start and end , section 13 , P266 . And see too AL Maqrizi , AL-Siluok , section 1 , p544 . AL-Maqrizi , AL-Siluok , section1 , p544 . IbnKatheer , Start and end , section 13 , P266 . And see too AL Maqrizi , AL-Siluok , section 1 , p544 . AL M
i i
AL Sil
k
i
1
544 IbnKatheer , Start and end , section 13 , P266 . And see too AL Maqrizi , AL-Siluok , section 1 , p544 . AL-Maqrizi , AL-Siluok , section1 , p544 . The same source , section 1 , p.p 544-545 . And see too AL-Yonini , ThailMeraat AL-Zaman , section 2 , p156 ;
AL-Nowairi , Nihayat AL-Erb , section 30 , p63 ; IbnAebik AL-Dawadari , Kanz AL-Durar , section8 ,
p.p 90-91 . Al-Qalqashandi , AL-khutat , section 2 , p637 . IbnShadad , History of King AL-Zaher , p.p337-338 . AL-Maqrizi , AL-Siluok , section 1 , p561 . And see too IbnAbd Al-zaher , AL-RawdAlzaher , p137 ; Bibars
AL-Mansuri , zubdat AL-Fikra , p 84-85 ; AL-Nowairi , Nihayat AL-Erb , section 31 , p38 ; ALthahabi ,
History of Islam , section 49 , p 8 ; AL-Yafuaai , ThailMeraatALzaman , section 4 , p121 . AL-Maqrizi , AL-Siluok , section1, p561 . And see too Al-Yonini ,ThailMeraat AL-Zaman , section 2 , p195
.Shiraz : It's an ancient city in the western south of BiladFaris , Mohammed Ibn Al-khasimrepuit it by the
time of the Islamic conquest (695AH/713AD) , Shiraz means a hollow of lion. when the Islamic militry
arrived to BiladFaris , they dewlt in that place and settled there till Astakhar city established , MuslIms
were pleased with this place and built Shiraz . the city was valuable , well improved , it was full of
binding , there were no walls , markets, houses had big garden of fruits . Yaqut AL-Hamawi
,MuaajamALBoldan , section3 , p 381 ; AL-Humairy , ALRawd AL-Zahir , p351 . (Saif AL-DeenBiklik , he was one of the Mogol princes who arrived to Egypt during the reign of The sultan
Bibars, The sultan anpowered him and crowned him as a prince of AL-Tabilkhanat in Cairo . Author Information
Prof. Abdul Khaliq Khames Ali (Ph.D.)
Department of History, College of Education for
HumanitiesUniversity of Diyala,Iraq
Zainab Adnan Hashem
Department of History, College of Education for
HumanitiesUniversity of Diyala,Iraq References AL-
Nowairi , Nihayat AL-Erb , section 30 , p99 ; AL-Maqrizi , AL-Siluok , section2 , p 8 . (Khafaja : It's one of the most important trip of AkeelIbnKab Bin Amir Bin Saasaan . Their name comes from
their grandfather Khafaja Bin Amro Bin Akeen Bin kaab .Orginally they were an Iraqi princes inhabited
Heat and AL-Anbar till Nakhla and Kufa . The most prominent figures of this trips are Klider Bin Badran
Bin Muqalid Bin Suluman Bin Mharish AL-Ebadi ,Shahir Bin Ahmed AL-Khafagi .IbnFadlullah AL
Omari , Msalik AL Absar , section 4 , p.p351-352 ; Al-Qalqashandi , Nehayat AL-Erb Fe MaarifatAnsab
Al-Arab , p246 ; And Qalaaed AL Juman , p122-123 . AL-Maqrizi , AL-Siluok , section 2 , p8 . And see too AL-Nowairi ,Nehayat AL-Erb , section 30 , p99 . The same source section2
p10 AL-Maqrizi , AL-Siluok , section 2 , p8 . And see too AL-Nowairi ,Nehayat AL-Erb , section 30 , p
The same source , section2 , p10 . The same source , section2 , p10 . The same source , section2 , p10 . AL-Luoq : Linguistically , in Arabic languages AL-Luoq means soften things. It located in the front part of
Cairo. The ground of this city was fertile a since water of Nile had receded there . Al-Qalqashandi ,
Suboh AL-Aasha , section3 , p408 ; AL-Maqrizi , Al-Khutat , section3 , p210 ; Dahman , Muaajam AL-
Alfaz AL-Tarikhia , p133 . AL-Maqrizi , Al-Khutat , section3 , p209 . Al-Qalqashandi ,Suboh AL-Aasha , section3 , p408 . AL-Maqrizi , AL-Siluok , section2 , p11 . The prince Shams AL-DeenBahider : He is Shams AL-DeenBahiderIbnFaraj bin Abdulla Sahib Samisat , The
prince arrived to Cairo in (672AH/1274AD) and the sultan Bibars empowered him, he stayed in Cairo
till his death in (676AH/1278AD) . IbnTahgriBardy , AL-Dalil AL-Shafi , section1 , p199 . Maqrizi , AL-Siluok , section2 , p86 ; AL-Nowairi , Nehayat AL-Erb , section30 , p207 . AL-Maqrizi , AL-Siluok , section2 , p86 ; AL-Nowairi , Nehayat AL-Erb , section30 , p207 . AL-Maqrizi , AL-Siluok , section2 , p86 . IbnAbd Al-Zaher , History of the king Al-Zaher , p.p337-338 . IbnAbd Al-Zaher , History of the king Al-Zaher , p.p337-338 . The same source , section2 , p86 . 24 The same source , section2 , p86 . References 24 Nawoar , Communities of Mongol , p35 . AL Maqrizi , AL Siluok , section2 , p49 . The same source , section2 , p165 . IbnKatheer , Start and end , section14 , p40 . Author Information
Prof. Abdul Khaliq Khames Ali (Ph.D.)
Department of History, College of Education for
HumanitiesUniversity of Diyala,Iraq
Zainab Adnan Hashem
Department of History, College of Education for
HumanitiesUniversity of Diyala,Iraq 325
|
https://openalex.org/W4362533291
|
https://figshare.com/articles/journal_contribution/Supplemental_Table_2_from_Race-associated_Molecular_Changes_in_Gynecologic_Malignancies/22544586/1/files/40008000.pdf
|
English
| null |
Supplemental Table 4 from Race-associated Molecular Changes in Gynecologic Malignancies
| null | 2,023
|
cc-by
| 4,128
|
Supplemental Table 2 Supplemental Table 2 Gene
logFC
adj.P.Val
Tumor
GSTM1
3.54
1.07E-03
CESC
HBB
1.78
4.65E-02
CESC
GSTT2
1.22
4.65E-02
CESC
IHH
2.46
3.86E-02
UCEC
PDIA2
1.63
4.82E-02
UCEC
HSD17B3
1.51
6.23E-03
UCEC
GPR83
1.37
1.37E-02
UCEC
LRGUK
1.22
3.36E-02
UCEC
C3orf32
1.17
1.58E-02
UCEC
NOTCH2NL
1.14
2.54E-10
UCEC
LRRIQ3
1.03
4.64E-02
UCEC
DISP2
-1.16
5.88E-03
UCEC
C21orf56
-1.30
3.80E-02
UCEC
RPS28
-2.49
8.28E-03
UCEC
ULK4
1.18
7.82E-08
OV
PLCH1
2.03
2.42E-07
OV
ACSM3
1.34
1.53E-05
OV
CEACAM21
-2.03
1.53E-05
OV
CD300C
-1.81
4.05E-05
OV
EFR3B
1.34
4.40E-05
OV
C1QC
-1.51
4.93E-05
OV
SLC26A7
1.99
4.93E-05
OV
BSN
1.28
5.09E-05
OV
ARHGDIB
-1.15
5.09E-05
OV
C1QB
-1.55
7.39E-05
OV
TYROBP
-1.41
9.70E-05
OV
C3orf15
1.20
1.03E-04
OV
FGFRL1
1.21
1.47E-04
OV
GPSM3
-1.33
1.54E-04
OV
C1QA
-1.45
1.71E-04
OV
TBX1
1.85
2.05E-04
OV
ZNF389
1.22
2.24E-04
OV
PDLIM2
-1.02
2.60E-04
OV
GYPC
-1.26
2.77E-04
OV
LAPTM5
-1.31
2.90E-04
OV
TGFBI
-1.23
3.84E-04
OV
HLA-DPA1
-1.51
4.42E-04
OV
FILIP1
1.37
4.44E-04
OV CRYBB1
-1.38
5.03E-04
OV
HNMT
-1.10
5.03E-04
OV
HS3ST1
-1.21
5.03E-04
OV
CD68
-1.12
5.27E-04
OV
MDH1B
1.01
5.27E-04
OV
SRGN
-1.15
5.59E-04
OV
MSI1
1.57
5.73E-04
OV
SIGLEC9
-1.49
6.33E-04
OV
SLCO2B1
-1.42
6.33E-04
OV
AIF1
-1.52
6.34E-04
OV
FCER1G
-1.28
6.44E-04
OV
C2orf39
2.04
6.91E-04
OV
ZNF334
1.19
6.93E-04
OV
ZNF443
1.04
6.93E-04
OV
CRIP3
1.49
7.19E-04
OV
DNAH7
1.25
7.19E-04
OV
DNAH10
1.27
7.37E-04
OV
SLC46A3
-1.04
8.02E-04
OV
HYDIN
1.13
8.52E-04
OV
MAF
-1.20
8.58E-04
OV
DNAH6
1.45
8.87E-04
OV
GLRX
-1.00
8.87E-04
OV
MS4A4A
-1.50
8.90E-04
OV
LOC100233209
-1.69
9.43E-04
OV
CD14
-1.18
9.93E-04
OV
C17orf60
-1.56
1.00E-03
OV
LRRC37A2
-1.17
1.00E-03
OV
GIMAP6
-1.03
1.27E-03
OV
ADORA3
-1.52
1.29E-03
OV
CYBB
-1.51
1.35E-03
OV
LSP1
-1.15
1.35E-03
OV
HAMP
-1.81
1.35E-03
OV
RNASE2
-1.50
1.38E-03
OV
CLEC18A
1.57
1.48E-03
OV
NPL
-1.03
1.49E-03
OV
C1orf162
-1.20
1.50E-03
OV
RNASE6
-1.41
1.53E-03
OV
C20orf26
1.56
1.58E-03
OV
LILRA2
-1.45
1.58E-03
OV
LRP4
1.44
1.58E-03
OV
MS4A6A
-1.50
1.58E-03
OV RCSD1
-1.05
1.58E-03
OV
DCN
-1.49
1.61E-03
OV
TBXAS1
-1.31
1.68E-03
OV
LRRIQ3
1.03
1.71E-03
OV
CACNB4
1.04
1.78E-03
OV
CCDC74B
1.02
1.82E-03
OV
MGC16121
-1.59
1.82E-03
OV
ADAP2
-1.01
1.84E-03
OV
GNG2
-1.01
1.85E-03
OV
HLA-DPB1
-1.33
1.85E-03
OV
GMFG
-1.07
1.91E-03
OV
APOBEC3H
-1.44
1.94E-03
OV
DSCAML1
1.63
2.06E-03
OV
GIMAP4
-1.11
2.06E-03
OV
MSR1
-1.24
2.07E-03
OV
FOLR2
-1.25
2.07E-03
OV
PIK3CG
-1.39
2.07E-03
OV
CD300A
-1.19
2.12E-03
OV
IRF8
-1.44
2.12E-03
OV
PDIA2
1.82
2.19E-03
OV
APOC2
-2.01
2.24E-03
OV
C1S
-1.18
2.24E-03
OV
FCGR2A
-1.14
2.24E-03
OV
GAB3
-1.12
2.24E-03
OV
ABLIM3
-1.11
2.24E-03
OV
EFEMP1
-1.89
2.28E-03
OV
ITGB2
-1.24
2.28E-03
OV
DNAH11
1.60
2.39E-03
OV
CD52
-1.56
2.40E-03
OV
LAIR1
-1.36
2.57E-03
OV
SELPLG
-1.16
2.66E-03
OV
S1PR4
-1.20
2.72E-03
OV
LY86
-1.29
2.72E-03
OV
MT2A
-1.06
2.75E-03
OV
HKDC1
1.23
2.75E-03
OV
CRABP2
-1.16
2.76E-03
OV
RSPH10B2
1.35
2.77E-03
OV
CSF1R
-1.22
2.77E-03
OV
VSIG4
-1.43
2.81E-03
OV
ALOX5AP
-1.69
2.85E-03
OV
SIGLEC7
-1.18
2.87E-03
OV PPARG
-1.34
2.89E-03
OV
HAVCR2
-1.32
2.90E-03
OV
HOPX
-1.29
2.95E-03
OV
RNF157
1.08
2.95E-03
OV
GIMAP1
-1.00
3.02E-03
OV
TTLL9
1.19
3.03E-03
OV
GPR34
-1.39
3.07E-03
OV
F13A1
-1.52
3.10E-03
OV
WDR65
1.46
3.29E-03
OV
CLEC18B
1.53
3.32E-03
OV
MS4A7
-1.15
3.39E-03
OV
TMEM176A
-1.54
3.59E-03
OV
CMKLR1
-1.17
3.66E-03
OV
HLA-DMA
-1.18
3.66E-03
OV
HCST
-1.06
3.73E-03
OV
LRRC25
-1.12
3.82E-03
OV
LY96
-1.22
3.87E-03
OV
CTSW
-1.90
3.93E-03
OV
JAK3
-1.01
3.93E-03
OV
SIRPB2
-1.44
3.93E-03
OV
SLC38A3
1.34
3.93E-03
OV
GAL3ST4
-1.01
4.01E-03
OV
GNG7
-1.35
4.01E-03
OV
CD300LF
-1.41
4.04E-03
OV
CXCL12
-1.65
4.15E-03
OV
CD163
-1.34
4.20E-03
OV
SIGLEC11
-1.84
4.20E-03
OV
TNFSF8
-1.48
4.22E-03
OV
FCGR3A
-1.24
4.26E-03
OV
PSTPIP1
-1.20
4.42E-03
OV
TUBB2B
2.14
4.42E-03
OV
ARHGAP9
-1.31
4.43E-03
OV
FERMT3
-1.09
4.48E-03
OV
CD37
-1.12
4.55E-03
OV
CSF3R
-1.54
4.55E-03
OV
PRDM8
-1.19
4.58E-03
OV
CSF2RA
-1.44
4.58E-03
OV
KLRB1
-2.02
4.58E-03
OV
PIK3R5
-1.05
4.64E-03
OV
RASGRP4
-1.19
4.64E-03
OV
LST1
-1.37
4.73E-03
OV HIST1H4C
-1.12
4.91E-03
OV
SPI1
-1.07
4.91E-03
OV
BTK
-1.35
5.19E-03
OV
WAS
-1.03
5.20E-03
OV
CYTH4
-1.17
5.29E-03
OV
CA14
1.10
5.38E-03
OV
ABI3
-1.05
5.39E-03
OV
LYZ
-1.56
5.39E-03
OV
GIMAP7
-1.02
5.47E-03
OV
TLR4
-1.07
5.49E-03
OV
P2RY13
-1.47
5.57E-03
OV
DOCK8
-1.12
5.62E-03
OV
CD86
-1.27
5.72E-03
OV
LAT2
-1.16
5.80E-03
OV
VENTX
-1.25
5.83E-03
OV
SIGLEC5
-1.29
5.86E-03
OV
TMEM232
1.36
5.90E-03
OV
HPGDS
-1.27
5.93E-03
OV
C17orf87
-1.07
5.93E-03
OV
KLK10
-1.52
5.93E-03
OV
SELM
-1.28
5.93E-03
OV
SPP1
-1.05
5.93E-03
OV
CD53
-1.21
6.03E-03
OV
CD7
-1.62
6.07E-03
OV
MRC1
-1.38
6.19E-03
OV
BIN2
-1.24
6.24E-03
OV
CD247
-1.40
6.32E-03
OV
PTPRCAP
-1.22
6.38E-03
OV
EVI2B
-1.32
6.38E-03
OV
C3AR1
-1.29
6.41E-03
OV
CLIC5
-1.21
6.41E-03
OV
DOK2
-1.07
6.49E-03
OV
KIAA0748
-1.44
6.49E-03
OV
SASH3
-1.26
6.49E-03
OV
RIPK3
-1.05
6.54E-03
OV
TREM2
-1.33
6.88E-03
OV
CASC1
1.20
7.12E-03
OV
NCKAP1L
-1.28
7.19E-03
OV
ZNF727
1.35
7.19E-03
OV
GPR77
-1.09
7.24E-03
OV
SLA
-1.14
7.24E-03
OV NCF4
-1.04
7.28E-03
OV
VAV1
-1.16
7.33E-03
OV
DLEC1
1.22
7.52E-03
OV
DPYD
-1.33
7.52E-03
OV
TNFAIP8L2
-1.10
7.52E-03
OV
TNFRSF11A
-1.15
7.52E-03
OV
TLR8
-1.50
7.62E-03
OV
SALL1
1.68
7.65E-03
OV
SLC9A9
-1.04
7.65E-03
OV
IL10RA
-1.15
7.92E-03
OV
PLEK
-1.18
7.96E-03
OV
NKX3-1
-1.34
8.05E-03
OV
FCGR1B
-1.16
8.13E-03
OV
WBSCR26
-2.24
8.15E-03
OV
C10orf128
-1.23
8.27E-03
OV
FCRLB
-1.32
8.27E-03
OV
FGL2
-1.27
8.27E-03
OV
TMEM200A
-1.18
8.31E-03
OV
DARC
-1.60
8.35E-03
OV
FLRT1
1.22
8.37E-03
OV
HK3
-1.24
8.52E-03
OV
SNORA12
-1.39
8.52E-03
OV
NFAM1
-1.08
8.57E-03
OV
MGP
-1.03
8.65E-03
OV
FPR1
-1.40
8.81E-03
OV
PRSS35
-1.59
8.87E-03
OV
APOC1
-1.08
8.88E-03
OV
FCGR2C
-1.48
8.92E-03
OV
HCK
-1.13
8.92E-03
OV
SERPINA1
-1.25
8.92E-03
OV
CLDN11
-1.27
8.94E-03
OV
SAMSN1
-1.16
8.96E-03
OV
RTN1
-1.22
9.10E-03
OV
TFPI2
-1.90
9.10E-03
OV
APBB1IP
-1.29
9.19E-03
OV
COL9A3
1.48
9.30E-03
OV
PTPN7
-1.08
9.42E-03
OV
PRSS33
2.14
9.46E-03
OV
TMEM176B
-1.21
9.46E-03
OV
TMEM150B
-1.32
9.51E-03
OV
C16orf54
-1.23
9.52E-03
OV LOC441666
1.07
9.60E-03
OV
NCF2
-1.04
9.68E-03
OV
PTCH2
1.01
9.72E-03
OV
LRRIQ1
1.17
9.77E-03
OV
C13orf33
-1.55
1.02E-02
OV
C7orf58
-1.05
1.02E-02
OV
SDS
-1.09
1.02E-02
OV
EPSTI1
-1.11
1.03E-02
OV
IGF1
-1.27
1.03E-02
OV
RERG
-1.53
1.04E-02
OV
DKK2
-1.25
1.04E-02
OV
DNAH2
1.29
1.04E-02
OV
LOC221442
1.10
1.04E-02
OV
IL12RB1
-1.42
1.04E-02
OV
EVI2A
-1.28
1.07E-02
OV
TLR7
-1.27
1.07E-02
OV
MYO1F
-1.05
1.08E-02
OV
MATK
-1.49
1.08E-02
OV
CLEC7A
-1.27
1.12E-02
OV
AOAH
-1.46
1.12E-02
OV
IL32
-1.12
1.13E-02
OV
P2RY8
-1.04
1.13E-02
OV
CD180
-1.10
1.18E-02
OV
CYTIP
-1.05
1.19E-02
OV
LGR5
1.64
1.19E-02
OV
ITGAM
-1.28
1.19E-02
OV
SAMD3
-1.30
1.19E-02
OV
FCRL6
-1.67
1.20E-02
OV
WDFY4
-1.34
1.21E-02
OV
SLC38A5
-1.65
1.22E-02
OV
AMH
-2.00
1.23E-02
OV
MARCO
-2.01
1.23E-02
OV
CD6
-1.17
1.24E-02
OV
MPEG1
-1.07
1.24E-02
OV
FCGR1A
-1.20
1.25E-02
OV
GPR65
-1.09
1.25E-02
OV
HGFAC
1.23
1.25E-02
OV
TNFSF13B
-1.18
1.26E-02
OV
LOC349196
-1.05
1.26E-02
OV
RASAL3
-1.10
1.26E-02
OV
RASL11A
-1.28
1.26E-02
OV GRAP2
-1.43
1.29E-02
OV
CD33
-1.03
1.30E-02
OV
C14orf49
-1.12
1.30E-02
OV
EMILIN3
1.15
1.31E-02
OV
APOL3
-1.18
1.32E-02
OV
PARP15
-1.20
1.32E-02
OV
SGSM1
1.24
1.33E-02
OV
CD209
-1.17
1.34E-02
OV
PTAFR
-1.03
1.36E-02
OV
CLEC2B
-1.02
1.36E-02
OV
MAN1A1
-1.13
1.41E-02
OV
FCGR1C
-1.13
1.41E-02
OV
C12orf68
-1.02
1.42E-02
OV
RGS18
-1.12
1.43E-02
OV
CD3G
-1.47
1.44E-02
OV
CD84
-1.25
1.44E-02
OV
LILRB4
-1.24
1.44E-02
OV
SERPINF1
-1.24
1.46E-02
OV
TTC9
-1.15
1.46E-02
OV
LOC285830
-1.02
1.47E-02
OV
GAPT
-1.30
1.47E-02
OV
CYSLTR1
-1.18
1.52E-02
OV
GDF1
1.07
1.52E-02
OV
GFAP
1.45
1.52E-02
OV
SIT1
-1.39
1.53E-02
OV
IL2RA
-1.25
1.53E-02
OV
PDGFRA
-1.44
1.54E-02
OV
PTPN22
-1.24
1.54E-02
OV
DOCK2
-1.24
1.60E-02
OV
TAGAP
-1.03
1.61E-02
OV
KCNMB2
1.11
1.62E-02
OV
WNK2
1.01
1.63E-02
OV
ALDH1A1
-1.04
1.64E-02
OV
CD48
-1.28
1.64E-02
OV
HLA-DRA
-1.01
1.64E-02
OV
CLEC10A
-1.68
1.69E-02
OV
SLC22A3
-1.46
1.69E-02
OV
GNLY
-1.98
1.72E-02
OV
PROM1
1.56
1.73E-02
OV
LRRK2
-1.10
1.73E-02
OV
KLHL4
-1.27
1.75E-02
OV SLAMF8
-1.24
1.75E-02
OV
SSPO
1.12
1.76E-02
OV
ANKRD22
-1.30
1.78E-02
OV
KRT15
-1.67
1.78E-02
OV
COPZ2
-1.15
1.78E-02
OV
SLC24A3
-1.30
1.78E-02
OV
FOLR3
-2.21
1.79E-02
OV
NTN4
-1.03
1.79E-02
OV
C12orf59
-1.17
1.80E-02
OV
LILRB5
-1.08
1.81E-02
OV
GPR82
-1.30
1.84E-02
OV
HSD11B1
-1.23
1.85E-02
OV
TSIX
1.14
1.85E-02
OV
C2orf40
1.20
1.90E-02
OV
ARHGAP15
-1.19
1.95E-02
OV
CCR2
-1.54
1.97E-02
OV
MEI1
-1.33
1.97E-02
OV
FPR3
-1.18
1.99E-02
OV
CFD
-1.07
2.01E-02
OV
FCGR2B
-1.41
2.02E-02
OV
PTPRC
-1.19
2.02E-02
OV
SLAMF6
-1.40
2.03E-02
OV
RARRES3
-1.11
2.03E-02
OV
VCAM1
-1.41
2.07E-02
OV
CHGA
1.60
2.10E-02
OV
LEFTY2
1.44
2.12E-02
OV
CAMK2B
1.09
2.14E-02
OV
FAM81B
1.60
2.14E-02
OV
SNAI2
-1.19
2.14E-02
OV
VTCN1
-1.58
2.14E-02
OV
ARMC3
1.16
2.15E-02
OV
NCF1C
-1.17
2.18E-02
OV
IKZF1
-1.01
2.20E-02
OV
TMEM215
-1.65
2.20E-02
OV
LCN2
-1.38
2.22E-02
OV
CXCR3
-1.37
2.22E-02
OV
FYB
-1.16
2.23E-02
OV
GPR120
-1.24
2.25E-02
OV
KYNU
-1.03
2.25E-02
OV
LILRB2
-1.07
2.26E-02
OV
SLC2A5
-1.05
2.28E-02
OV RASGRF2
-1.01
2.30E-02
OV
GPR84
-1.20
2.30E-02
OV
ARHGAP20
-1.19
2.33E-02
OV
GZMK
-1.85
2.33E-02
OV
AMICA1
-1.20
2.34E-02
OV
SNX20
-1.02
2.34E-02
OV
CCR5
-1.20
2.34E-02
OV
FMO2
-2.27
2.39E-02
OV
RUNX1T1
-1.44
2.41E-02
OV
S100A3
-1.19
2.42E-02
OV
DDX11L2
-1.42
2.47E-02
OV
PDCD1LG2
-1.18
2.47E-02
OV
CPM
-1.06
2.48E-02
OV
SORCS2
-1.83
2.49E-02
OV
B3GNT3
-1.32
2.51E-02
OV
LRRC16B
1.11
2.59E-02
OV
CBFA2T3
-1.07
2.59E-02
OV
CD5
-1.21
2.60E-02
OV
LOXL4
-1.20
2.72E-02
OV
TLR10
-1.18
2.72E-02
OV
TMEM71
-1.01
2.72E-02
OV
KLK1
-1.80
2.73E-02
OV
WISP1
-1.16
2.75E-02
OV
IL10
-1.04
2.75E-02
OV
SELL
-1.09
2.75E-02
OV
PDLIM3
-1.17
2.79E-02
OV
KLRK1
-1.50
2.81E-02
OV
SIGLEC10
-1.16
2.83E-02
OV
SECTM1
-1.17
2.84E-02
OV
KLK13
-1.45
2.86E-02
OV
FOXQ1
-1.44
2.88E-02
OV
GZMA
-1.55
2.89E-02
OV
CA8
1.22
2.90E-02
OV
ADAM33
1.06
2.92E-02
OV
DPEP1
-1.68
2.94E-02
OV
BCAT1
1.31
2.95E-02
OV
PLA2G2A
-2.29
3.02E-02
OV
HGF
-1.03
3.08E-02
OV
LTA
-1.06
3.10E-02
OV
TFAP2C
-1.12
3.10E-02
OV
MNDA
-1.03
3.13E-02
OV DEFB1
-2.38
3.14E-02
OV
IGSF21
-1.20
3.14E-02
OV
CSF2RB
-1.06
3.14E-02
OV
DNAI1
1.34
3.16E-02
OV
SLA2
-1.08
3.17E-02
OV
RGS6
-1.15
3.21E-02
OV
CCL3
-1.02
3.23E-02
OV
PRPH
1.10
3.25E-02
OV
OSR1
-1.30
3.26E-02
OV
DIRAS1
-1.49
3.27E-02
OV
TSPAN7
1.05
3.32E-02
OV
CLEC12A
-1.32
3.38E-02
OV
ATP1A3
-2.06
3.40E-02
OV
ANXA8L2
-1.67
3.41E-02
OV
AIM2
-1.38
3.43E-02
OV
SLC22A18AS
-1.11
3.43E-02
OV
SFRP4
-1.67
3.44E-02
OV
CHRNB4
-1.19
3.47E-02
OV
MGAT5B
1.59
3.50E-02
OV
ADAMTS8
1.01
3.50E-02
OV
RHD
1.13
3.53E-02
OV
FGFR3
1.05
3.57E-02
OV
PDZK1IP1
-1.52
3.61E-02
OV
RGS4
-1.10
3.64E-02
OV
VWA3A
1.45
3.73E-02
OV
CCL5
-1.31
3.77E-02
OV
PRF1
-1.11
3.77E-02
OV
ATP1B2
1.07
3.79E-02
OV
CLDN10
-2.19
3.82E-02
OV
CALML5
-2.30
3.84E-02
OV
BTC
-1.12
3.85E-02
OV
CRTAM
-1.21
3.86E-02
OV
CD96
-1.03
3.87E-02
OV
TIMP4
-1.39
3.89E-02
OV
HLA-DQA1
-1.33
3.93E-02
OV
XKR9
-1.31
3.94E-02
OV
EFCAB4B
-1.02
3.95E-02
OV
UBASH3A
-1.23
3.97E-02
OV
IL2RG
-1.19
3.98E-02
OV
STEAP4
-1.34
4.01E-02
OV
LUM
-1.30
4.02E-02
OV TNIK
-1.31
4.03E-02
OV
CEACAM6
-2.08
4.06E-02
OV
PTPRN
-1.21
4.06E-02
OV
IBSP
-1.52
4.08E-02
OV
ITK
-1.37
4.12E-02
OV
CD2
-1.53
4.18E-02
OV
SLC10A4
1.02
4.18E-02
OV
LYVE1
-1.04
4.18E-02
OV
MSC
-1.27
4.18E-02
OV
IL21R
-1.69
4.22E-02
OV
FASLG
-1.25
4.24E-02
OV
XCL1
-1.13
4.27E-02
OV
NTRK2
-1.12
4.29E-02
OV
TCF21
-1.63
4.40E-02
OV
CLEC5A
-1.07
4.47E-02
OV
OSM
-1.05
4.47E-02
OV
FBN3
1.40
4.48E-02
OV
MEIS3
-1.12
4.50E-02
OV
IL2RB
-1.24
4.63E-02
OV
KRT4
-2.18
4.64E-02
OV
KLK7
-1.00
4.64E-02
OV
CD8B
-1.31
4.65E-02
OV
GPR1
-1.19
4.65E-02
OV
ITGB6
-1.43
4.70E-02
OV
CLEC4E
-1.34
4.71E-02
OV
THSD7A
1.01
4.73E-02
OV
PREX2
1.01
4.75E-02
OV
CCL14
-1.34
4.84E-02
OV
EBI3
-1.01
4.90E-02
OV
IMPG2
1.52
4.98E-02
OV
A2ML1
1.29
1.47E-05
BRCA
ABAT
-1.08
1.47E-10
BRCA
ABCA12
-1.61
2.98E-07
BRCA
ABCA6
-1.23
3.81E-11
BRCA
ABCA8
-1.24
2.07E-05
BRCA
ABCA9
-1.29
3.85E-09
BRCA
ABCC11
-1.33
2.76E-04
BRCA
ABCC6
-1.10
8.29E-09
BRCA
ABCC8
-1.41
2.47E-05
BRCA
ABCC9
-1.02
1.52E-13
BRCA
ABCG2
-1.06
1.20E-14
BRCA ACCN3
1.01
5.56E-14
BRCA
ACER2
-1.27
3.38E-20
BRCA
ACOX2
-1.10
8.74E-08
BRCA
ACVR1C
-1.02
5.32E-06
BRCA
ADAM22
-1.26
2.22E-15
BRCA
ADAMTS15
-1.68
1.17E-11
BRCA
ADAMTS18
-1.06
9.76E-07
BRCA
ADAT3
1.02
1.69E-25
BRCA
ADCK5
1.02
1.88E-36
BRCA
ADCY5
-1.35
2.28E-07
BRCA
ADH1B
-1.86
2.19E-04
BRCA
ADIPOQ
-2.13
3.07E-05
BRCA
AFF3
-1.38
2.49E-09
BRCA
AGER
1.04
3.22E-28
BRCA
AGR2
-1.48
4.41E-06
BRCA
AGR3
-2.01
3.90E-05
BRCA
AGTR1
-1.47
2.33E-07
BRCA
AIM1L
1.04
4.00E-09
BRCA
AK5
-1.32
2.14E-07
BRCA
AKAP12
-1.02
2.26E-12
BRCA
AKAP2
-1.08
6.67E-12
BRCA
ALOX12P2
1.11
1.07E-06
BRCA
AMPH
-1.07
3.73E-10
BRCA
ANGPTL1
-1.20
1.46E-10
BRCA
ANGPTL6
1.05
9.30E-17
BRCA
ANKRD30A
-2.91
2.97E-08
BRCA
ANKRD30B
-1.24
1.28E-04
BRCA
ANKRD43
-1.22
4.53E-07
BRCA
ANO3
-1.33
4.49E-07
BRCA
ANO5
-1.18
2.77E-07
BRCA
AQP7
-1.13
2.52E-04
BRCA
AQPEP
-1.00
4.22E-06
BRCA
ARHGAP20
-1.18
5.08E-14
BRCA
ARHGEF38
-1.20
2.55E-10
BRCA
AR
-2.15
6.63E-14
BRCA
ASPA
-1.11
1.50E-09
BRCA
ATAD3B
1.04
6.56E-35
BRCA
ATP1A2
-1.15
2.65E-05
BRCA
ATP1A4
-1.89
2.59E-15
BRCA
AURKB
1.41
6.67E-26
BRCA
AVPR1A
-1.28
3.08E-17
BRCA B3GALT5
-1.14
3.68E-06
BRCA
B3GNT4
1.08
2.22E-13
BRCA
BAIAP2L2
1.33
2.08E-20
BRCA
BARX1
1.28
8.79E-10
BRCA
BIRC5
1.26
3.10E-18
BRCA
C14orf73
1.26
1.35E-15
BRCA
C16orf59
1.02
8.34E-21
BRCA
C16orf79
1.21
1.75E-25
BRCA
C18orf56
1.14
3.59E-22
BRCA
C19orf20
1.25
2.83E-23
BRCA
C19orf60
1.09
7.41E-40
BRCA
C1orf106
1.01
1.73E-07
BRCA
C20orf151
1.01
8.04E-19
BRCA
C21orf70
1.05
1.21E-39
BRCA
C2CD4D
1.13
8.16E-26
BRCA
C2orf48
1.13
5.48E-12
BRCA
C4orf48
1.43
1.87E-24
BRCA
C6orf108
1.10
7.41E-40
BRCA
C6orf223
1.10
4.59E-07
BRCA
C8ORFK29
1.07
1.83E-15
BRCA
CALML5
1.52
8.00E-08
BRCA
CAPS
1.21
7.38E-18
BRCA
CBS
1.15
7.45E-14
BRCA
CBX2
1.01
8.55E-13
BRCA
CCDC154
1.09
3.02E-15
BRCA
CCDC42B
1.09
8.63E-14
BRCA
CCDC78
1.03
8.24E-10
BRCA
CCDC85B
1.04
1.37E-26
BRCA
CCDC88B
1.00
2.08E-25
BRCA
CCL3L1
1.19
2.00E-15
BRCA
CCNE1
1.12
1.19E-17
BRCA
CD19
1.17
6.51E-07
BRCA
CDC20
1.17
1.45E-17
BRCA
CDC45
1.12
2.00E-16
BRCA
CDCA3
1.02
1.91E-18
BRCA
CDKN2A
1.37
1.07E-23
BRCA
CDT1
1.24
5.97E-28
BRCA
CENPA
1.05
3.05E-15
BRCA
CENPM
1.09
1.53E-21
BRCA
CHODL
1.13
2.07E-08
BRCA
CHTF18
1.07
5.39E-31
BRCA CLDN3
1.02
2.25E-11
BRCA
CLEC18A
1.14
3.56E-20
BRCA
CLEC18B
1.10
2.13E-14
BRCA
CLIC3
1.07
7.13E-13
BRCA
COL11A2
1.43
5.77E-17
BRCA
COL2A1
1.14
3.77E-04
BRCA
COL9A3
1.55
1.04E-15
BRCA
COMTD1
1.17
1.79E-32
BRCA
CRABP1
1.12
2.60E-04
BRCA
CRLF1
1.01
1.08E-09
BRCA
CYP2D7P1
1.03
8.60E-19
BRCA
DDN
1.09
1.44E-12
BRCA
DGCR5
1.09
1.02E-12
BRCA
DNER
1.04
1.15E-05
BRCA
DUSP9
1.04
2.36E-06
BRCA
E2F1
1.02
1.46E-20
BRCA
EMID2
1.09
1.98E-10
BRCA
EN1
1.32
1.35E-12
BRCA
ENHO
1.24
6.65E-15
BRCA
ESPNL
1.10
2.15E-12
BRCA
FAM131C
1.35
1.01E-15
BRCA
FAM64A
1.06
4.54E-16
BRCA
FBN3
1.09
4.29E-06
BRCA
FER1L4
1.04
2.60E-13
BRCA
FOXD1
1.20
7.10E-10
BRCA
FOXH1
1.25
2.20E-20
BRCA
FUT7
1.15
7.21E-18
BRCA
FZD9
1.21
2.50E-10
BRCA
GAL
1.40
5.56E-11
BRCA
GFRA3
1.05
4.28E-07
BRCA
GJB3
1.56
4.27E-14
BRCA
GJB5
1.62
2.52E-16
BRCA
GLDC
1.21
2.37E-07
BRCA
GLI4
1.11
6.11E-37
BRCA
GPC2
1.30
3.06E-20
BRCA
GSDMC
1.16
5.75E-10
BRCA
GSTM1
2.26
1.57E-11
BRCA
HAGHL
1.21
9.39E-17
BRCA
HES4
1.41
2.44E-34
BRCA
HIST1H2AM
1.04
6.28E-11
BRCA
HIST1H2BO
1.09
2.65E-11
BRCA HIST2H3C
1.02
8.28E-12
BRCA
HIST3H2A
1.08
6.66E-11
BRCA
HOMER3
1.04
5.30E-37
BRCA
HPDL
1.21
7.67E-14
BRCA
IER5L
1.10
1.83E-29
BRCA
ISG15
1.29
2.42E-16
BRCA
JSRP1
1.85
2.68E-22
BRCA
KCNG1
1.48
7.19E-12
BRCA
KIF18B
1.16
5.81E-17
BRCA
KIF1A
1.04
7.01E-04
BRCA
KIFC2
1.03
7.96E-25
BRCA
KRT16
1.70
6.55E-11
BRCA
KRT6A
1.21
7.26E-05
BRCA
KRT6B
1.08
4.39E-04
BRCA
KRT6C
1.41
6.44E-08
BRCA
KRT81
1.61
1.11E-10
BRCA
KRT86
1.10
1.08E-09
BRCA
LCN2
1.01
7.38E-05
BRCA
LGALS4
1.22
2.19E-17
BRCA
LGALS7
1.02
5.97E-06
BRCA
LIME1
1.26
1.31E-24
BRCA
LOC339535
1.15
5.33E-07
BRCA
LOC400696
1.36
4.96E-12
BRCA
LTB
1.21
1.15E-13
BRCA
LYG1
1.03
6.15E-21
BRCA
MEF2B
1.28
1.13E-28
BRCA
MIA
1.14
1.33E-05
BRCA
MIF
1.08
7.65E-25
BRCA
MND1
1.04
5.09E-14
BRCA
MSLN
2.43
1.82E-18
BRCA
MST1P9
1.15
9.17E-15
BRCA
MYBL2
1.28
1.51E-15
BRCA
MYLK2
1.32
1.59E-20
BRCA
NACA2
1.08
8.95E-35
BRCA
NCRNA00105
1.02
3.41E-20
BRCA
NFKBIL2
1.01
1.01E-25
BRCA
NOTUM
1.30
1.79E-12
BRCA
NOXO1
1.14
6.32E-15
BRCA
NPM2
1.05
1.95E-09
BRCA
NRTN
1.34
6.88E-17
BRCA
NSUN5P1
1.11
2.61E-39
BRCA ORC6L
1.01
2.46E-18
BRCA
P2RY11
1.59
9.52E-15
BRCA
PAQR6
1.05
9.54E-19
BRCA
PCSK1N
2.18
1.12E-19
BRCA
PIF1
1.55
3.09E-39
BRCA
PITX1
1.00
8.57E-07
BRCA
PKMYT1
1.44
1.04E-22
BRCA
POMC
1.07
4.31E-15
BRCA
PPP1R14C
1.07
1.95E-06
BRCA
PRAME
1.46
2.76E-05
BRCA
PRR22
1.00
2.28E-23
BRCA
PRR7
1.30
1.53E-26
BRCA
PRSS50
1.23
6.44E-10
BRCA
RAD54L
1.06
4.87E-18
BRCA
RBP1
1.15
8.73E-16
BRCA
RCOR2
1.09
1.52E-13
BRCA
RECQL4
1.16
1.69E-24
BRCA
RFPL3S
1.14
1.79E-25
BRCA
RLTPR
1.21
1.15E-13
BRCA
RNF112
1.01
9.57E-17
BRCA
ROPN1B
1.18
1.64E-05
BRCA
RUFY4
1.36
2.00E-16
BRCA
SCAND1
1.01
9.69E-33
BRCA
SCT
1.05
4.16E-08
BRCA
SH2B2
1.00
2.55E-22
BRCA
SH3D20
1.10
6.22E-19
BRCA
SPC24
1.16
1.76E-14
BRCA
SPIB
1.08
1.22E-07
BRCA
SYCE1L
1.65
2.48E-18
BRCA
SYT8
1.36
2.51E-07
BRCA
TCAP
1.26
1.16E-16
BRCA
TMEM105
1.05
7.59E-09
BRCA
TMEM191A
1.05
7.85E-26
BRCA
TMPRSS5
1.12
3.04E-13
BRCA
TNFRSF25
1.13
3.82E-27
BRCA
TNFRSF4
1.08
8.77E-26
BRCA
TNFRSF6B
1.01
2.12E-23
BRCA
TNNI2
1.03
7.64E-11
BRCA
TNNT1
1.42
9.17E-08
BRCA
TPRXL
1.07
4.23E-09
BRCA
TREX2
1.05
9.30E-17
BRCA TROAP
1.20
6.50E-19
BRCA
TSIX
1.68
1.96E-17
BRCA
TSPAN10
1.21
1.46E-22
BRCA
TSPO
1.02
4.28E-38
BRCA
TUBB8
1.21
1.72E-24
BRCA
UBE2C
1.24
2.17E-16
BRCA
UBE2S
1.12
1.52E-30
BRCA
UPK3BL
1.11
5.26E-13
BRCA
UPK3B
1.34
2.04E-18
BRCA
VGF
1.75
6.32E-18
BRCA
VGLL1
1.12
4.87E-04
BRCA
WNT6
1.60
2.15E-15
BRCA
YJEFN3
1.09
9.42E-20
BRCA
ZNHIT2
1.03
6.80E-08
BRCA
BAGE2
-1.07
1.69E-11
BRCA
BCAS1
-1.08
1.10E-04
BRCA
BICC1
-1.40
1.56E-14
BRCA
BMPR1B
-1.10
2.26E-03
BRCA
BPIL1
-1.25
4.10E-04
BRCA
BTBD8
-1.25
1.11E-15
BRCA
C1QTNF7
-1.26
3.91E-11
BRCA
C1orf150
-1.28
5.07E-13
BRCA
C1orf173
-1.12
5.71E-05
BRCA
C1orf64
-1.95
4.13E-06
BRCA
C20orf114
-1.12
2.13E-03
BRCA
C20orf94
-1.02
4.23E-16
BRCA
C4orf31
-1.34
1.64E-12
BRCA
C6orf97
-1.13
8.76E-09
BRCA
C6
-1.11
1.23E-04
BRCA
C7orf58
-1.20
3.80E-13
BRCA
C8orf34
-1.03
1.50E-07
BRCA
CA12
-1.26
2.44E-10
BRCA
CACNA2D1
-2.11
1.08E-19
BRCA
CADM2
-1.08
1.86E-06
BRCA
CAPN8
-1.12
4.29E-04
BRCA
CASQ1
-1.11
1.09E-09
BRCA
CASQ2
-1.02
2.46E-07
BRCA
CCDC144A
-1.20
4.19E-11
BRCA
CCDC158
-1.44
7.79E-13
BRCA
CCDC160
-1.12
1.00E-09
BRCA
CCNT1
-1.36
1.97E-19
BRCA CD36
-1.24
2.62E-07
BRCA
CDKL5
-1.32
9.50E-14
BRCA
CEACAM5
-1.16
3.20E-03
BRCA
CES1
-1.36
7.63E-08
BRCA
CHAD
-1.20
9.73E-05
BRCA
CHL1
-1.35
5.93E-09
BRCA
CIDEA
-1.30
3.35E-04
BRCA
CIDEC
-1.18
1.32E-03
BRCA
CLGN
-1.06
1.58E-05
BRCA
CLIC6
-1.46
2.77E-06
BRCA
CLSTN2
-1.35
6.03E-09
BRCA
CNTN1
-1.44
5.12E-11
BRCA
CNTN4
-1.14
3.86E-09
BRCA
CNTNAP2
-1.16
3.34E-04
BRCA
COL14A1
-1.13
6.56E-12
BRCA
CPA3
-1.08
2.26E-06
BRCA
CPB1
-1.96
4.58E-04
BRCA
CSRNP3
-1.25
6.35E-12
BRCA
CST1
-1.16
2.63E-03
BRCA
CST2
-1.55
1.20E-08
BRCA
CT62
-1.30
9.98E-08
BRCA
CTTNBP2
-1.15
2.08E-10
BRCA
CX3CR1
-1.08
3.14E-10
BRCA
CYP2A6
-1.36
2.00E-04
BRCA
CYP2B7P1
-1.77
1.02E-04
BRCA
CYP4B1
-1.07
1.54E-04
BRCA
CYP4X1
-1.25
4.86E-06
BRCA
CYP4Z1
-2.06
3.26E-06
BRCA
CYP4Z2P
-1.67
9.24E-07
BRCA
DACH1
-1.28
1.28E-07
BRCA
DCDC2
-1.12
2.74E-05
BRCA
DCLK1
-1.25
1.57E-11
BRCA
DDI2
-1.18
5.59E-17
BRCA
DDR2
-1.11
6.53E-11
BRCA
DEFB132
-1.33
1.28E-06
BRCA
DNAH5
-1.22
8.09E-09
BRCA
DNAH7
-1.13
1.77E-13
BRCA
DOC2B
-1.04
1.24E-07
BRCA
DYNC2H1
-1.18
1.70E-21
BRCA
EBF2
-1.20
2.97E-12
BRCA
EDIL3
-1.49
5.24E-13
BRCA ENTPD3
-1.07
3.05E-10
BRCA
EPHA3
-1.03
5.57E-10
BRCA
ERBB4
-1.99
3.83E-10
BRCA
ESR1
-1.81
3.12E-12
BRCA
EYA4
-1.10
2.85E-07
BRCA
F13A1
-1.01
1.74E-10
BRCA
F7
-1.22
1.84E-06
BRCA
FABP4
-1.17
2.20E-03
BRCA
FAIM2
-1.05
1.18E-07
BRCA
FAM106C
-1.16
3.64E-13
BRCA
FAM184B
-1.29
4.60E-07
BRCA
FAM196B
-1.10
4.65E-06
BRCA
FAM198B
-1.09
1.05E-19
BRCA
FAM63B
-1.04
3.44E-19
BRCA
FAT3
-1.59
3.02E-20
BRCA
FAT4
-1.09
7.59E-14
BRCA
FGF10
-1.80
2.13E-10
BRCA
FHL5
-1.17
5.80E-13
BRCA
FLRT3
-1.09
6.93E-07
BRCA
FMN1
-1.21
1.47E-14
BRCA
FMO2
-1.02
2.18E-05
BRCA
FOXA1
-1.47
7.00E-09
BRCA
FOXP2
-1.08
8.68E-09
BRCA
GABRB3
-1.67
1.55E-10
BRCA
GALNT5
-1.09
2.31E-08
BRCA
GCNT4
-1.10
1.74E-12
BRCA
GFRA1
-1.96
5.84E-11
BRCA
GLDN
-1.01
4.49E-09
BRCA
GLRB
-1.10
2.99E-11
BRCA
GP2
-2.24
1.76E-07
BRCA
GPD1
-1.06
1.83E-03
BRCA
GPR81
-1.42
3.74E-12
BRCA
GPR98
-1.09
1.25E-04
BRCA
GRIK3
-1.07
7.58E-05
BRCA
GRPR
-1.99
4.99E-10
BRCA
GTF2A1
-1.12
4.42E-20
BRCA
GUSBP3
-1.03
5.86E-15
BRCA
HEPACAM2
-1.47
1.21E-06
BRCA
HIPK3
-1.21
1.73E-20
BRCA
HMCN1
-1.03
1.30E-11
BRCA
HMGCS2
-2.16
4.36E-06
BRCA HRASLS5
-1.19
7.84E-06
BRCA
HRNR
-1.08
1.71E-11
BRCA
HS6ST3
-1.17
8.11E-05
BRCA
HTR2B
-1.04
1.36E-16
BRCA
IGF1
-1.03
5.11E-08
BRCA
IGSF9B
-1.20
4.49E-09
BRCA
IL20RA
-1.59
3.07E-12
BRCA
IL33
-1.05
1.10E-05
BRCA
IL6ST
-2.07
2.54E-23
BRCA
INPP4B
-1.05
8.24E-11
BRCA
IPMK
-1.04
2.01E-14
BRCA
ITGA8
-1.17
1.04E-10
BRCA
ITPR1
-1.08
1.60E-18
BRCA
KCND3
-1.77
1.87E-14
BRCA
KCNJ3
-1.62
1.20E-04
BRCA
KERA
-1.10
1.26E-08
BRCA
KIAA0754
-1.16
9.30E-12
BRCA
KIAA1244
-1.08
9.50E-13
BRCA
KIAA1377
-1.08
4.13E-13
BRCA
KLHDC7A
-1.47
1.59E-05
BRCA
KLHL11
-1.17
4.68E-19
BRCA
KL
-1.18
2.15E-15
BRCA
LAMA2
-1.10
2.48E-11
BRCA
LCOR
-1.17
2.71E-18
BRCA
LEP
-1.31
5.43E-04
BRCA
LGALS12
-1.21
2.70E-05
BRCA
LNPEP
-1.12
6.26E-20
BRCA
LOC100272216
-1.07
2.30E-15
BRCA
LOC145837
-1.07
7.66E-06
BRCA
LOC148696
-1.34
1.27E-14
BRCA
LOC728606
-1.14
1.20E-04
BRCA
LONRF2
-1.31
5.89E-09
BRCA
LPHN3
-1.09
2.66E-08
BRCA
LRP1B
-1.25
3.54E-05
BRCA
LRRC31
-2.30
1.10E-10
BRCA
LRRC37A2
-1.13
3.39E-34
BRCA
LUZP2
-1.09
1.17E-06
BRCA
MAN2A1
-1.04
1.82E-19
BRCA
MAOA
-1.10
1.54E-06
BRCA
MAST4
-1.09
1.10E-14
BRCA
MBNL3
-1.02
3.68E-11
BRCA MGAT5
-1.07
5.47E-13
BRCA
MICALCL
-1.16
1.15E-13
BRCA
MKX
-1.09
5.65E-06
BRCA
MMRN1
-1.18
6.35E-07
BRCA
MS4A2
-1.21
1.55E-09
BRCA
MUC6
-1.01
9.81E-03
BRCA
MYH11
-1.07
4.21E-07
BRCA
MYO9A
-1.03
1.88E-19
BRCA
MYOCD
-1.26
3.66E-12
BRCA
MYRIP
-1.46
3.03E-13
BRCA
N4BP2
-1.04
1.10E-15
BRCA
NAT1
-1.44
8.08E-08
BRCA
NAV3
-1.04
1.42E-09
BRCA
NBEAL1
-1.24
2.49E-17
BRCA
NBEA
-1.14
9.14E-15
BRCA
NCAM2
-1.42
4.21E-10
BRCA
NCOA2
-1.02
2.86E-13
BRCA
NEK10
-1.44
8.16E-07
BRCA
NEK5
-1.03
4.62E-08
BRCA
NELL2
-1.09
1.29E-05
BRCA
NFATC2
-1.17
3.10E-12
BRCA
NHSL2
-1.35
7.51E-22
BRCA
NOVA1
-1.29
3.59E-07
BRCA
NRXN3
-1.03
9.37E-10
BRCA
NTN4
-1.17
4.64E-12
BRCA
NTRK2
-1.15
2.30E-07
BRCA
ODZ2
-1.05
1.97E-06
BRCA
OGN
-1.12
3.97E-05
BRCA
OMD
-1.07
7.09E-08
BRCA
P2RY12
-1.01
1.53E-08
BRCA
P2RY1
-1.00
4.00E-16
BRCA
PAR5
-1.42
1.17E-14
BRCA
PARD3B
-1.22
1.06E-18
BRCA
PCDHA4
-1.01
1.22E-05
BRCA
PCDHA6
-1.17
2.73E-07
BRCA
PCDHGA2
-1.07
8.77E-10
BRCA
PCDHGA4
-1.02
6.36E-10
BRCA
PCDHGA9
-1.00
1.63E-11
BRCA
PCDHGB7
-1.18
1.31E-13
BRCA
PCK1
-1.42
4.08E-05
BRCA
PDE11A
-1.15
1.54E-10
BRCA PDE3A
-1.22
1.71E-13
BRCA
PDE8B
-1.07
1.44E-08
BRCA
PDK4
-1.15
9.16E-09
BRCA
PGR
-1.81
2.87E-07
BRCA
PIP
-1.68
9.07E-04
BRCA
PIWIL2
-1.07
1.24E-09
BRCA
PLCXD3
-1.10
4.41E-06
BRCA
PLIN1
-1.26
3.93E-04
BRCA
PLXNC1
-1.19
4.38E-13
BRCA
PPAN-P2RY11
-1.56
1.32E-05
BRCA
PPFIA2
-1.01
3.94E-11
BRCA
PREX2
-1.84
6.25E-19
BRCA
PRG4
-1.01
7.84E-05
BRCA
PRKG1
-1.40
2.14E-17
BRCA
PRKG2
-1.42
6.08E-16
BRCA
PRND
-1.04
2.15E-09
BRCA
PRSS21
-1.65
9.28E-09
BRCA
PTPRT
-1.76
7.92E-07
BRCA
RAB27B
-1.29
5.36E-15
BRCA
RAB30
-1.14
4.47E-15
BRCA
RALGAPA2
-1.31
1.00E-19
BRCA
RANBP3L
-1.13
9.79E-07
BRCA
RAPGEF6
-1.10
2.72E-17
BRCA
RASEF
-1.38
2.75E-11
BRCA
RASSF6
-1.05
1.01E-07
BRCA
RBMS3
-1.03
4.09E-13
BRCA
RERGL
-1.08
7.62E-05
BRCA
REST
-1.02
4.12E-16
BRCA
RGPD5
-1.12
7.83E-15
BRCA
RGS22
-1.56
1.18E-08
BRCA
RGS7BP
-1.25
4.61E-11
BRCA
ROBO2
-1.51
9.57E-08
BRCA
RORB
-1.06
2.93E-09
BRCA
RPS28
-1.49
5.49E-06
BRCA
RPS6KA6
-1.05
6.03E-08
BRCA
RUNDC2A
-1.03
5.87E-20
BRCA
RUNX1T1
-1.22
1.70E-14
BRCA
SCGB1D2
-1.62
1.29E-03
BRCA
SCGB2A2
-2.15
1.80E-04
BRCA
SCN2B
-1.28
2.61E-08
BRCA
SCN3A
-1.11
5.59E-08
BRCA SCN7A
-2.12
5.02E-17
BRCA
SCUBE2
-1.74
1.04E-11
BRCA
SEMA3D
-1.04
1.13E-07
BRCA
SEMA3E
-1.42
1.86E-08
BRCA
SEMA5A
-1.07
7.95E-17
BRCA
SERINC5
-1.23
1.30E-15
BRCA
SERPINA11
-1.70
2.23E-06
BRCA
SLC13A2
-1.01
8.22E-04
BRCA
SLC14A2
-1.38
1.86E-11
BRCA
SLC16A12
-1.28
4.81E-10
BRCA
SLC16A6
-1.08
9.01E-07
BRCA
SLC16A7
-1.26
1.57E-13
BRCA
SLC18A2
-1.23
4.11E-10
BRCA
SLC24A2
-1.00
3.23E-05
BRCA
SLC30A8
-1.28
1.21E-03
BRCA
SLC38A11
-1.16
2.48E-12
BRCA
SLC40A1
-1.29
8.09E-16
BRCA
SLC44A4
-1.37
1.18E-06
BRCA
SLC44A5
-1.09
2.36E-06
BRCA
SLC4A4
-1.05
8.98E-07
BRCA
SLC7A2
-1.62
7.22E-11
BRCA
SLIT2
-1.29
6.73E-14
BRCA
SLITRK6
-1.43
2.18E-05
BRCA
SNORD116-4
-1.38
1.59E-08
BRCA
SORCS1
-1.48
2.94E-06
BRCA
SOX5
-1.09
4.91E-10
BRCA
SPATA18
-1.12
1.88E-12
BRCA
ST8SIA6
-1.82
2.40E-14
BRCA
STK32B
-1.11
4.96E-08
BRCA
SVEP1
-1.19
2.19E-10
BRCA
SYNPO2
-1.00
4.37E-11
BRCA
SYT1
-1.16
1.83E-05
BRCA
SYT9
-1.74
1.19E-07
BRCA
SYTL5
-1.26
1.46E-05
BRCA
TAOK1
-1.22
1.32E-15
BRCA
TAT
-1.07
8.33E-03
BRCA
TBC1D9
-1.07
7.91E-13
BRCA
TDRD1
-1.11
3.31E-05
BRCA
TFAP2B
-1.65
3.32E-04
BRCA
TFF1
-2.18
8.78E-05
BRCA
TFF3
-1.28
2.23E-04
BRCA TGFBR3
-1.10
8.06E-10
BRCA
THRSP
-1.22
3.26E-05
BRCA
THSD4
-1.02
5.41E-09
BRCA
THSD7A
-1.04
1.43E-13
BRCA
THSD7B
-1.25
7.84E-11
BRCA
TIMP4
-1.18
1.44E-05
BRCA
TLL1
-1.17
1.73E-13
BRCA
TMC5
-1.78
5.29E-09
BRCA
TMEM132C
-1.01
1.40E-03
BRCA
TMTC1
-1.12
1.20E-09
BRCA
TNIK
-1.05
3.26E-11
BRCA
TNNI3K
-1.05
7.90E-10
BRCA
TOX3
-1.21
8.13E-06
BRCA
TP63
-1.01
9.09E-05
BRCA
TPRG1
-1.19
9.57E-06
BRCA
TRIM58
-1.02
3.72E-06
BRCA
TSHZ2
-1.23
1.68E-12
BRCA
TTBK2
-1.14
1.05E-22
BRCA
TUSC5
-1.68
3.39E-05
BRCA
UGCG
-1.12
1.49E-13
BRCA
UHMK1
-1.20
1.53E-14
BRCA
UNC5C
-1.10
6.91E-12
BRCA
VWA2
-1.23
2.57E-07
BRCA
WDR65
-1.29
1.11E-11
BRCA
ZBTB16
-1.38
4.43E-07
BRCA
ZFHX4
-1.05
8.77E-14
BRCA
ZNF192
-1.13
6.82E-16
BRCA
ZNF214
-1.14
4.85E-14
BRCA
ZNF354C
-1.34
1.28E-18
BRCA
ZNF366
-1.15
1.86E-15
BRCA
ZNF676
-1.24
7.01E-09
BRCA
ZNF699
-1.16
7.90E-20
BRCA
ZNF727
-1.07
6.75E-08
BRCA LogFC AA vs EA; Log2 ( mean EA tumor RPKM) / (mean AA tumor RPKM) for each
transcript LogFC AA vs EA; Log2 ( mean EA tumor RPKM) / (mean AA tumor RPKM) for each
transcript adj.P.Val: P value adjusted for multiple hypothesis testing with BH method Tumor: CESC = cervix, UCEC = uterine endometrial carcinoma, OV = ovary, BRCA =
breast
|
https://openalex.org/W4213286143
|
https://www.frontiersin.org/articles/10.3389/fonc.2022.821958/pdf
|
English
| null |
Transcriptional Expressions of ALDH1A1/B1 as Independent Indicators for the Survival of Thyroid Cancer Patients
|
Frontiers in oncology
| 2,022
|
cc-by
| 7,556
|
ORIGINAL RESEARCH
published: 23 February 2022
doi: 10.3389/fonc.2022.821958 Methods: We used UALCAN, Human Protein Atlas, Kaplan–Meier plotter, TISIDB,
TIMER, and KOBAS databases to investigate the expression and role of ALDH1 in
thyroid cancer progression. In addition, quantitative real-time polymerase chain reaction
was performed to detect the expression of the target genes in thyroid cancer cell lines and
cancer tissues. Edited by:
Yun Dai,
First Affiliated Hospital of Jilin
University, China
Reviewed by:
Tao Yi,
Hong Kong Baptist University, Hong
Kong, SAR China
Lianjun Zhang,
Suzhou Institute of Systems Medicine
(ISM), China
*Correspondence: Edited by:
Yun Dai,
First Affiliated Hospital of Jilin
University, China Reviewed by:
Tao Yi,
Hong Kong Baptist University, Hong
Kong, SAR China
Lianjun Zhang,
Suzhou Institute of Systems Medicine
(ISM), China Results: Expression of ALDH1A1/B1 was significantly decreased based on individual
cancer stages and tumor histology, and high levels of ALDH1A1/B1 were associated with
poor overall survival in thyroid cancer patients. Moreover, ALDH1A1/B1 expression was
negatively correlated with immune-stimulating genes, major histocompatibility complex,
chemokines, and receptors. *Correspondence:
Gang Wu
yingwu2002@163.com Conclusions: These results suggest that ALDH1A1/B1 might serve as potential
prognostic biomarkers for thyroid cancer diagnosis. Specialty section:
This article was submitted to
Cancer Molecular Targets
and Therapeutics,
a section of the journal
Frontiers in Oncology
Received: 25 November 2021
Accepted: 24 January 2022
Published: 23 February 2022 Keywords: ALDH1, thyroid cancer, prognostic biomarker, tumor-infiltrating lymphocytes, cancer progression Transcriptional Expressions of
ALDH1A1/B1 as Independent
Indicators for the Survival of
Thyroid Cancer Patients
Ying Cui 1, Yao Liu 1, Lan Mu 1, Yang Li 1 and Gang Wu 2*
1 Department of Otorhinolaryngology, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, China,
2 Department of Hepatobiliary Surgery, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, China 1 Department of Otorhinolaryngology, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, China,
2 Department of Hepatobiliary Surgery, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, China Background: Aldehyde dehydrogenase (ALDH) 1 is an important enzyme involved in the
regulation of several cellular mechanisms via aldehyde detoxification. High ALDH1 levels
were correlated with tumorigenesis and stemness maintenance in cancer. INTRODUCTION Thyroid cancer is the most prevalent endocrine malignancy worldwide, with a global incidence of
over 500,000 per year (1). The number of endocrine cases has been growing rapidly for decades (2),
resulting in approximately 90% of the endocrine malignancies and 70% of deaths (3). Thyroid
cancer was reported to be prevalent in women (4), but more advanced-stage cases were found in
men (5). Recently, different research groups have explored prognostic markers and therapeutic
targets for thyroid cancer, and some progress has been made (1, 4, 6). However, current diagnosis of
thyroid cancer is still far from sufficient owing to the lack of appropriate biomarkers. Received: 25 November 2021
Accepted: 24 January 2022
Published: 23 February 2022 Survival Analysis Initiation and progression of many cancers could be
characterized by aberrant change of gene expression (17). Here, we used Kaplan–Meier plotter (http://kmplot.com/
analysis/) to access the prognostic value of ALDH1 level in
thyroid cancer patients. Kaplan–Meier plotter can investigate
the gene expression level in pan-caner under TCGA consortium
support (18). We investigated expression levels of target genes in
thyroid cancer tissues and corresponding adjacent tissues, and
the search term was limited to thyroid cancer. Finally, we
analyzed data based on patient’s gender to plot the OS, first
progression (FP), and post progression survival (PPS) of patients
with thyroid cancer, with the hazard ratio (HR) (95% confidence
intervals) and log rank P-value. In this study, we systematically investigated the role of the
ALDH1 family in thyroid cancer patients by searching public
databases. We demonstrated that the levels of ALDH1A1/B1 had
been significantly decreased in thyroid cancer patients. Aberrantly elevated ALDH1A1/B1 levels in cancer tissues were
negatively correlated with OS in thyroid cancer patients. Besides,
the perturbation of ALDH1A1/A3/B1 expression was
significantly correlated with pathological stage. Finally, we
elucidated the important but overlooked role of ALDH1A1/B1
in tumor-infiltrating lymphocytes (TILs), and we revealed a non-
negligible immune regulatory role of ALDH1A1/B1 by drawing
gene expression heat maps. Overall, our data provide motivation
to better understand the tumor promotion and immune
inhibition role of ALDH1A1/B1 in thyroid cancer patients,
suggesting that ALDH1A1/B1 might serve as potential
prognostic biomarkers for thyroid cancer treatment. Analysis of Target Genes in Tumor-
Infiltrating Immune Cells Tumor IMmune Estimation Resource (TIMER, https://cistrome. shinyapps.io/timer/) contains 10,897 tumors from 32 cancer
types (19), which could be used to systematically analyze the
association between the target genes and tumor-infiltrating
immune cells. Here, we investigated the somatic copy number
alterations of target genes with the Somatic copy number
alterations
(SCNA) module. And correlation between target
genes and infiltrating immune cells also was analyzed by limiting
the search term to thyroid cancer. In addition, the correlation between target genes with
lymphocytes and immunomodulators was investigated by
using the TISIDB database (http://cis.hku.hk/TISIDB/
index.php). Citation: Recent studies demonstrated that Nuclear factor E2-related
factor 2 (Nrf2) activation is involved in cancer stem cell-like
properties’ exposure to high ALDH1A1 concentrations (9) that
result in shorter overall survival (OS) and progression-free
survival in ovarian cancer patients (10). In addition, targeting
ALDH1 may facilitate the elimination of cancer stem cells and
inhibit breast cancer progression (11, 12). Cumulative evidence
has revealed that aberrant expression of ALDH1 is being
evaluated as a potential novel prognostic marker in breast
cancer (13), head and neck squamous cell carcinoma (7),
ovarian cancer (9), and lung cancer (14). However, the role of
the ALDH1 family in thyroid cancer remains poorly understood. Construction of Protein–Protein
Interaction The Gene Expression Profiling Interactive Analysis (GEPIA)
dataset is a web-based tool exploited by Tang et al. (15) to
analyze the RNA sequencing (RNA-seq) expression data in
cancer tissues and normal samples. Here, we used GEPIA to
investigate the target genes’ expression level in thyroid cancer
tissues and normal control and analyzed the relationship
between target genes and pathological stage in thyroid cancer
patients. And search term was limited to thyroid cancer. In
addition, target proteins expressed in thyroid cancer tissues were
confirmed by immunohistochemistry data acquired from The
Human Protein Atlas (https://www.proteinatlas.org/). inBio Discover (https://inbio-discover.com/) could visualize
protein–protein interaction networks under genome scale in
humans (20). inBio Discover contains over 500,000
interactions from 8 source databases. Here, we used inBio
Discover to construct target gene networks by limiting the
search terms to ALH1A1, ALDH1A3, and ALDH1B1. Cell Culture and Sample Collection
Thyroid epithelial cell line nthy-ori 3-1 and thyroid cancer cell
lines (CHO-K1 and TPC-1) were purchased from the
Shanghai Zhong Qiao Xin Zhou Biotechnology Co., Ltd. Cell lines were mycoplasma negative, and nthy-ori 3-1 was
cultured in RPMI 1640 medium, and thyroid cancer cell lines
were cultured in Dulbecco’s modified Eagle’s medium
(DMEM). All cell lines were supplemented with 10% fetal
bovine serum (Gibco, USA). Citation: Cui Y, Liu Y, Mu L, Li Y and Wu G
(2022) Transcriptional Expressions
of ALDH1A1/B1 as Independent
Indicators for the Survival of
Thyroid Cancer Patients. Front. Oncol. 12:821958. doi: 10.3389/fonc.2022.821958 The aldehyde dehydrogenase 1 (ALDH1) family consists of conserved enzymes that oxidize
aldehydes to carboxylic acids (7). The ALDH1 family includes six members: ALDH1A1 to
ALDH1A3, ALDH1B1, ALDH1L1, and ALDH1l2 (8), which are localized in cytosol and February 2022 | Volume 12 | Article 821958 Frontiers in Oncology | www.frontiersin.org Cui et al. ALDH1A1/B1 in Thyroid Cancer mitochondria and can be found in almost all important organs
(8). Recent studies demonstrated that Nuclear factor E2-related
factor 2 (Nrf2) activation is involved in cancer stem cell-like
properties’ exposure to high ALDH1A1 concentrations (9) that
result in shorter overall survival (OS) and progression-free
survival in ovarian cancer patients (10). In addition, targeting
ALDH1 may facilitate the elimination of cancer stem cells and
inhibit breast cancer progression (11, 12). Cumulative evidence
has revealed that aberrant expression of ALDH1 is being
evaluated as a potential novel prognostic marker in breast
cancer (13), head and neck squamous cell carcinoma (7),
ovarian cancer (9), and lung cancer (14). However, the role of
the ALDH1 family in thyroid cancer remains poorly understood. In this study, we systematically investigated the role of the
ALDH1 family in thyroid cancer patients by searching public
databases. We demonstrated that the levels of ALDH1A1/B1 had
been significantly decreased in thyroid cancer patients. Aberrantly elevated ALDH1A1/B1 levels in cancer tissues were
negatively correlated with OS in thyroid cancer patients. Besides,
the perturbation of ALDH1A1/A3/B1 expression was
significantly correlated with pathological stage. Finally, we
elucidated the important but overlooked role of ALDH1A1/B1
in tumor-infiltrating lymphocytes (TILs), and we revealed a non-
negligible immune regulatory role of ALDH1A1/B1 by drawing
gene expression heat maps. Overall, our data provide motivation
to better understand the tumor promotion and immune
inhibition role of ALDH1A1/B1 in thyroid cancer patients,
suggesting that ALDH1A1/B1 might serve as potential
prognostic biomarkers for thyroid cancer treatment. metastasis. P-value was determined based on Student’s t-test,
and P < 0.05 was considered as statistically significant. mitochondria and can be found in almost all important organs
(8). UALCAN UALCAN (http://ualcan.path.uab.edu) is a free web database
based on The Cancer Genome Atlas (TCGA) level 3 RNA-seq
and 31 different types of cancer (16). In the current study, we
investigated the expression level of target genes in thyroid cancer
tissues and corresponding adjacent tissues based on UALCAN
platform, and the search term was limited to thyroid cancer. Finally, we analyzed data based on individual cancer stages,
patient’s gender, patient’s age, tumor histology, and nodal Five thyroid cancer tissues and their corresponding adjacent
tissues were collected from the First Hospital of Jilin University
and stored at -80°C conditions. All participants were given
written informed consent, and the current study has been February 2022 | Volume 12 | Article 821958 Frontiers in Oncology | www.frontiersin.org 2 ALDH1A1/B1 in Thyroid Cancer Cui et al. approved by the ethics committee of the First Hospital of
Jilin University. t-test analysis was performed on GraphPad Prism 7.0, and P <
0.05 was considered statistically significant. RESULTS We used TransZol Up (Transgen, CN) to extract total RNA from the
cell lines and cancer tissues according to manufacturer’s instructions. cDNA was synthesized by using the Hifair® II 1st Strand cDNA
Synthesis Kit (Yeasen, CN) according to manufacturer’s instructions. Then, quantitative real-time PCR (qRT-PCR) was performed using a
Hieff® qPCR SYBR Green Master Mix (Yeasen, CN) according to the
manufacturer’s instructions. Here, b-actin was used as endogenous
control. The primer sequence sets were listed as follows: b-actin: F: 5′
TTCAACACCCCAGCCATG3′, R: 5′CCTCGTAGATGG
GCACAGT3′; ALDH1A1: F: 5′ CCGTGGCGTACTATG
GATGC3′, R: 5′GCAGCAGACGATCTCTTTCGAT3′; ALDH1B1:
F :
5 ′ A G A G T C T T A C G C C T T G G A C T T 3 ′ ,
R :
5 ′
GTCTTGCCATGCCACTTGTC3′. Aberrant Expression of the ALDH1 Family
in Thyroid Cancer Patients The members of the ALDH1 family have been proposed as
potential therapeutic targets of ovarian cancer (21) and have
shown the prognostic value in lung cancer (14) and breast
carcinomas (22). However, the role of the ALDH1 family in
thyroid cancer is still ill-defined. Here, we investigated the
expression of the ALDH1 family in thyroid cancer, and we
found that all ALDH1 family members could be detected in
thyroid cancer tissues at the transcriptional level (Figure 1A). Interestingly, the expressions of ALDH1A1 and ALDH1B1 were
higher in the normal thyroid tissues and decreased in the thyroid
cancer tissues (Figure 1A). In contrast, the level of ALDH1A3
was slightly increased in cancer tissues (Figure 1A). The low
expression levels of other genes in the thyroid tissues were Statistical Analysis Significance was determined with the either HR and P-values for
plotting survival curves. The independent-samples Student’s A
B
C
D
FIGURE 1 | Confirmation of the expression of ALDH1 family members in thyroid cancer tissues. (A) ALDH1A1/A3/B1 were the top 3 genes expressed in thyroid
cancer tissues (T, tumor tissues, n = 503; N, normal tissues, n = 59). (B–D) ALDH1A1/A3/B1 expression was confirmed by immunohistochemistry data from The
Human Protein Atlas. Data in panel (A) were collected from the Gene Expression Profiling Interactive Analysis (GEPIA) database. Data in panels (B–D) were collected
from The Human Protein Atlas database. A B
C
D
FIGURE 1 | Confirmation of the expression of ALDH1 family members in thyroid cancer tissues. (A) ALDH1A1/A3/B1 were the top 3 genes expressed in thyroid
cancer tissues (T, tumor tissues, n = 503; N, normal tissues, n = 59). (B–D) ALDH1A1/A3/B1 expression was confirmed by immunohistochemistry data from The
Human Protein Atlas. Data in panel (A) were collected from the Gene Expression Profiling Interactive Analysis (GEPIA) database. Data in panels (B–D) were collected
from The Human Protein Atlas database. B C B FIGURE 1 | Confirmation of the expression of ALDH1 family members in thyroid cancer tissues. (A) ALDH1A1/A3/B1 were the top 3 genes expressed in thyroid
cancer tissues (T, tumor tissues, n = 503; N, normal tissues, n = 59). (B–D) ALDH1A1/A3/B1 expression was confirmed by immunohistochemistry data from The
Human Protein Atlas. Data in panel (A) were collected from the Gene Expression Profiling Interactive Analysis (GEPIA) database. Data in panels (B–D) were collected
from The Human Protein Atlas database. February 2022 | Volume 12 | Article 821958 Frontiers in Oncology | www.frontiersin.org 3 Cui et al. ALDH1A1/B1 in Thyroid Cancer negligible. Furthermore, immunohistochemical results verified
that ALDH1A1, ALDH1A3, and ALDH1B1 were highly
expressed in thyroid cancer at the protein level (Figures 1B–
D), suggesting the role of ALDH1A1/A3/B1 in thyroid cancer
development. Therefore, we determined the expressions of
ALDH1A1/A3/B1 in thyroid cancer tissues. possible prognostic value of these enzymes particularly for male
patients with thyroid cancer. possible prognostic value of these enzymes particularly for male
patients with thyroid cancer. Expressions of ALDH1A1/A3/B1 Were
Correlated With the Pathological Stage in
Thyroid Cancer To determine if ALDH1A1/A3/B1 is associated with tumor
progression and clinical outcome of thyroid cancer, we
assessed the correlation between the expression of ALDH1A1/
A3/B1 and the pathological stage of the patients. Results showed
that there was a significant correlation between the expression of
ALDH1A1, ALDH1A3, and ALDH1B1 with the pathological
stage (Figures 3A–C). These results indicate that the aberrant
expression of ALDH1A1/A3/B1 may correlate with
tumorigenesis and development. Prognostic Value of mRNA Expression of
ALDH1A1/A3/B1 in Thyroid Cancer
Patients Furthermore, we assessed the prognostic significance of ALDH1A1/
A3/B1 in patients using Kaplan–Meier plotter analysis. As shown in
Figure 4, patients were divided into low- and high-risk groups
based on the expression levels of ALDH1A1/A3/B1. Our results
showed that changes in the expression level of ALDH1A1/A3 had
no influence over the OS of female patients with thyroid cancer
(Figures 4A, B, D, E); however, increased mRNA level of
ALDH1A1 impaired OS in male patients with thyroid cancer
(Figure 4C). Moreover, high mRNA expression of ALDH1A3
only enhanced OS in male patients with thyroid cancer
(Figure 4F). And high levels of ALDH1B1 also impaired OS in
patients with thyroid cancer (Figures 4G–I). Taken together, we
identified that ALDH1A1/A3/B1 in thyroid cancer patients were
significantly correlated with OS in male patients, supporting the ALDH1A1/A3/B1 Broadly Interacted With
Multiple Proteins p
Furthermore, to study the role of ALDH1A1/A3/B1 in thyroid
cancer, we constructed protein–protein interaction networks by
using the tool of inBio Discover. The results showed that
ALDH1A1/A3/B1 could interact with various proteins (Figure 6),
most of which were identified as enzymes. Besides, we showed that
ALDH1A1 and ALDH1B1 could interact with transcription factors
[CCAAT/enhancer-binding protein b (CEBPB) and CCHC-type
zinc finger nucleic acid-binding protein (CNBP), respectively] and
membrane-bound proteins [ATP-binding cassette subfamily C
member 6 (ABCC6) and mucin 1 (MUC1)], suggesting that the
aberrant changes of ALDH1A1/B1 may alter multiple protein
expressions and intracellular signal transduction. Indeed, our
Gene Ontology (GO) enrichment analysis revealed a close
relationship between ALDH1A1/A3/B1 and ethanol oxidation
(GO:0006069), as well as retinol metabolic process (GO:0042572),
which are known to be important in detoxification (8),
tumorigenesis (25), and thyroid tumor progression
(Supplementary Material) (26). Specifically, we found that
ALDH1A1/A3/B1 were enriched in thyroid hormone binding
(GO:0070324). Similarly, Kyoto Encyclopedia of Genes and
Genomes (KEGG) pathway analysis also showed that ALDH1A1/
A3/B1 were associated with metabolic pathways (Supplementary
Material). Collectively, these results indicate that ALDH1A1/A3/B1
may influence thyroid cancer via metabolism and interaction with
various enzymes and transcription factors. ALDH1A1 and ALDH1B1 Expression Tumor-infiltrating lymphocytes (TILs) are shaped by interactions
between different types of killer cell, regulatory cell, stroma cell, and
tumor cell in the tumor microenvironment (23). Recent studies
have shown that tumor progression or suppression is closely
associated with TIL response in the tumor microenvironment
(24). Thus, we investigated the relationship between ALDH1A1/
A3/B1 expression and TILs using TIMER. ALDH1A1/B1 showed
weak correlation with the six key types of immune cell in the tumor
microenvironment (Figures 5A, C). Meanwhile, a significant
correlation between ALDH1A3 expression and neutrophils, CD4+
T cells, dendritic cells, macrophages, and B cells (Figure 5B) was
observed. These results suggest that ALDH1A1/A3/B1 may affect
multiple cells in the tumor microenvironment that may participate
in the tumor immune response. y
As the expressions of ALDH1A1/A3/B1 were relatively high in
thyroid cancer tissues, we assessed the level of ALDH1A1/A3/B1
in thyroid tumors and normal tissues. Our results showed that
the transcriptional levels of ALDH1A1 and ALDH1B1
significantly decreased in thyroid cancer tissues, whereas that
of ALDH1A3 increased (Figures 2A–C). Next, we further
assessed the changes in ALDH1A1/A3/B1 based on different
classifications. As shown in Figure 2, mRNA expressions of
ALDH1A1 and ALDH1B1 significantly decreased in thyroid
cancer tissues compared to that in normal groups based on
individual cancer stages (Figures 2D–F), gender (Figures 2G–I),
tumor histology (Figures 2J–L), and nodal metastasis
(Figures 2M–O). Meanwhile, the levels of ALDH1A3
significantly increased (Figures 2B, E, H, K, N). These results
indicate that ALDH1A1/A3/B1 may be correlated with and may
play a significant role in thyroid cancer progression. Aberrant Levels of ALDH1A1/A3/B1 Are
Correlated With Immune Molecular
Regulation in Thyroid Cancer Violin plots showed the distribution
of ALDH1A1/A3/B1 [(A–C), respectively] mRNA expression correlated with tumor stage in thyroid cancer patients (Stage I, n = 284; stage II, n = 52; stage III, n =
112; stage IV, n = 55). Data were collected from the Gene Expression Profiling Interactive Analysis (GEPIA) database. cancer immunotherapy (27). Using the TISIDB database, we
demonstrated that ALDH1A1/A3/B1 had significant negative
correlations with immune-stimulating genes, such as CD276,
TMEM173, TNFSF9, CD70, TNFRSF25, and CD40 in thyroid
cancer (Figure 7A). Moreover, high levels of ALDH1A1/B1 were
also associated with restrained expression of major
histocompatibility complex (MHC) genes (HLA-G, HLA-B,
HLA-DOB, and TAP1) (Figure 7B). Notably, previous studies
have demonstrated that higher levels of immune stimulators and
MHC promote better therapeutic effect in cancer patients (28,
29). In addition, the expressions of ALDH1A1/B1 were found to
be negatively correlated with chemokines (CXCL5, CXCL16, and
CCL13; and CXCL14, respectively) (Figure 7C) and receptors
(CXCR2, CCR4, and CCR8, and CCR9, CCR10, respectively)
(Figure 7D). Therefore, ALDH1A1/B1 may be involved in
regulating the above immune molecules. cancer immunotherapy (27). Using the TISIDB database, we
demonstrated that ALDH1A1/A3/B1 had significant negative
correlations with immune-stimulating genes, such as CD276,
TMEM173, TNFSF9, CD70, TNFRSF25, and CD40 in thyroid
cancer (Figure 7A). Moreover, high levels of ALDH1A1/B1 were
also associated with restrained expression of major
histocompatibility complex (MHC) genes (HLA-G, HLA-B,
HLA-DOB, and TAP1) (Figure 7B). Notably, previous studies
have demonstrated that higher levels of immune stimulators and
MHC promote better therapeutic effect in cancer patients (28,
29). In addition, the expressions of ALDH1A1/B1 were found to
be negatively correlated with chemokines (CXCL5, CXCL16, and
CCL13; and CXCL14, respectively) (Figure 7C) and receptors
(CXCR2, CCR4, and CCR8, and CCR9, CCR10, respectively)
(Figure 7D). Therefore, ALDH1A1/B1 may be involved in
regulating the above immune molecules. this study. We observed that the regulatory functions of
ALDH1A1 and ALDH1B1 in thyroid cancer were involved in
poor outcome in patients and inhibition of TILs. ALDH1 has been reported to play a role in promoting triple-
negative breast cancer progression and enhanced drug resistance
in a Notch receptor 4 (NOTCH4)-dependent manner (32, 33). Ciccone et al. (31) observed that overexpression of ALDH1A1
was significantly associated with Vascular endothelial growth
factor (VEGF)-mediated angiogenesis by retinoic acid/ Hypoxia-
inducible factor (HIF)-1a signal. Similarly, a study on esophageal
cancer demonstrated that ALDH1 was positively correlated with
higher clinical stage and poor prognosis (34). Aberrant Levels of ALDH1A1/A3/B1 Are
Correlated With Immune Molecular
Regulation in Thyroid Cancer Here, we showed
that ALDH1A1/B1 expressions were significantly decreased in
thyroid cancer patients, but ALDH1A3 was elevated. However,
the level of ALDH1A3 had no significant changes. These results
indicated the different roles of ALDH1 members in thyroid
cancer progression. ALDH1A1 could regulate retinoid
signaling via the CEBPB to promote stemness maintenance in
cancer stem cells (35). In addition, ALDH1A1 also profoundly
affected acetaldehyde metabolism and drug resistance in
chemotherapy (8), suggesting that ALDH1A1 could broadly
regulate biological processes in cancer. Indeed, our results
showed that elevated ALDH1A1 was negatively correlated with
OS of thyroid cancer patients in male patients but was beneficial
to Relapse-free survival (RFS) in female patients; it indicated that
differently expressed gonadal hormones may also be involved in
ALDH1A1 regulation in thyroid cancer development. Aberrant Levels of ALDH1A1/A3/B1 Are
Correlated With Immune Molecular
Regulation in Thyroid Cancer Immune modulation is a critical factor that should be considered
in cancer therapy, as numerous genes play a role in response to February 2022 | Volume 12 | Article 821958 Frontiers in Oncology | www.frontiersin.org 4 Cui et al. ALDH1A1/B1 in Thyroid Cancer A
B
C
D
E
F
J
K
L
G
H
I
M
N
O
Expression of ALDH1A1/B1 was significantly decreased in thyroid cancer tissues. Differential expression of ALDH1A1 (A), ALDH1A3 (B), and ALDH
cancer tissues and normal tissues based on individual cancer stage (D–F), patient’s gender (G–I), histological subtype (J–L), and nodal metastasi
(N0, no regional lymph node metastasis; N1, metastases in 1–3 axillary lymph nodes; N2, metastases in 4–9 axillary lymph nodes; N3, metastases
illary lymph nodes). **P < 0.01; ****P < 0.0001. Data were collected from the UALCAN database. ns, No significance. C
F C
F
I B D E D
E H G I I I H H G G L J J
K
L L K O M N O M N FIGURE 2 | Expression of ALDH1A1/B1 was significantly decreased in thyroid cancer tissues. Differential expression of ALDH1A1 (A), ALDH1A3 (B), and ALDH1B1
(C) in thyroid cancer tissues and normal tissues based on individual cancer stage (D–F), patient’s gender (G–I), histological subtype (J–L), and nodal metastasis
status (M–O) (N0, no regional lymph node metastasis; N1, metastases in 1–3 axillary lymph nodes; N2, metastases in 4–9 axillary lymph nodes; N3, metastases in
10 or more axillary lymph nodes). **P < 0.01; ****P < 0.0001. Data were collected from the UALCAN database. ns, No significance. February 2022 | Volume 12 | Article 821958 Frontiers in Oncology | www.frontiersin.org 5 Cui et al. Cui et al. ALDH1A1/B1 in Thyroid Cancer A
B
C
FIGURE 3 | Correlation between differentially expressed ALDH1A1/A3/B1 and the pathological stage of thyroid cancer patients. Violin plots showed the distribution
of ALDH1A1/A3/B1 [(A–C), respectively] mRNA expression correlated with tumor stage in thyroid cancer patients (Stage I, n = 284; stage II, n = 52; stage III, n =
112; stage IV, n = 55). Data were collected from the Gene Expression Profiling Interactive Analysis (GEPIA) database. C A C B C FIGURE 3 | Correlation between differentially expressed ALDH1A1/A3/B1 and the pathological stage of thyroid cancer patients. ALDH1A1/B1 Expressions Significantly
Decreased in Thyroid Cancer Cell Line
and Cancer Tissues We have observed a decreased expression of ALDH1A1/B1 in
thyroid cancer tissues (Figures 1A–D), which was correlated
with poor survival of patients (Figures 4C, G, I). Thus, we aimed
to verify the levels of ALDH1A1/B1 in thyroid cancer cell lines
and thyroid cancer tissues in vitro. We found that ALDH1A1/B1
expression was significantly reduced in CHO-K1 and TPC-1 cells
(Figures 8A, B). In addition, we also detected a decreased
expression of ALDH1A1/B1 in thyroid cancer tissues
(Figures 8C, D). These results confirmed the effectiveness and
reliability of the results in bioinformatic analysis. Although the tumorigenic property of ALDH1A3 in cancer
has been observed (8, 36), it is suggested that upregulated
epithelial-to-mesenchymal transition and cancer stem cell-like
properties related genes, including ALDH13, may be involved in
the lymphatic spread of papillary thyroid microcarcinoma (37). However, another study found that administration of ALDH
inhibitor had no impact on proliferation or spherogenicity in
several thyroid cancer cell lines (38). Here, we showed that
aberrant expression of ALDH1A3 was not associated with OS
in women. Meanwhile, we noted that there was a significant
correlation between the expression of ALDH1A3 with the
pathological stage in patients. These data suggest that increased
ALDH1A3 may only affect alternative subtypes of thyroid DISCUSSION ALDH1 is a critical gene mediating the enzymatic detoxification
of aldehydes (8), and it is also required in cell proliferation (30)
and stem cell differentiation (31) in cancers. Although all six
ALDH1 members have been reported to be intracellularly
involved in various biological processes, ALDH1A1/A3/B1
were the highly expressed genes in thyroid cancer tissues in February 2022 | Volume 12 | Article 821958 Frontiers in Oncology | www.frontiersin.org 6 ALDH1A1/B1 in Thyroid Cancer Cui et al. Cui et al. A
B
C
D
E
F
G
H
I
FIGURE 4 | Kaplan–Meier plot revealing the correlation between ALDH1A1/A3/B1 level and overall survival (OS) in thyroid cancer patients. Patients were divided in
two groups based on ALDH1A1/A3/B1 level [(A, D, G), respectively], and Kaplan–Meier plot of ALDH1A1/A3/B1 in female patients (B, E, H) and male patients (C,
I) with thyroid cancer (total number of patients with thyroid cancer, n = 502; total number of female patients with thyroid cancer, n = 367; total number of male
patients with thyroid cancer, n = 135). Data were collected from the Kaplan–Meier plotter database. C A B B E F D E F E F D I I H G I H G FIGURE 4 | Kaplan–Meier plot revealing the correlation between ALDH1A1/A3/B1 level and overall survival (OS) in thyroid cancer patients. Patients were divided into
two groups based on ALDH1A1/A3/B1 level [(A, D, G), respectively], and Kaplan–Meier plot of ALDH1A1/A3/B1 in female patients (B, E, H) and male patients (C, F,
I) with thyroid cancer (total number of patients with thyroid cancer, n = 502; total number of female patients with thyroid cancer, n = 367; total number of male
patients with thyroid cancer, n = 135). Data were collected from the Kaplan–Meier plotter database. cancer. However, the role of ALDH1A3 in thyroid cancer
development still needs further study. suggesting a correlation between ALDH1B1 and androgen. Notably, a study in prostate cell lines showed that cells with
elevated ALDH1B1 expression frequently expressed androgen
receptor (41). In addition, a study in prostate cancer cell line
LNCaP reported that androgen dihydrotestosterone was involved
in aldehyde dehydrogenase regulation (42). Also, a high level of
aldehyde dehydrogenase promoted metastatic growths in stem-like
prostate cancer cells (43). Therefore, androgen receptor expression
may be involved in ALDH1B1-induced thyroid cancer progression. DISCUSSION Upregulation of ALDH1B1 by Kras is associated with pancreatic
lineage homeostasis and initiation/development of pancreatic ductal
adenocarcinoma, which are characteristics of enhanced production
of reactive oxygen species by altering mitochondrial metabolism
(39). ALDH1B1 is an enzyme that has been proposed to play an
important role in acetaldehyde detoxification. Increased ALDH1B1
level was involved in the regulation of the Wnt-, Notch-, and
Phosphatidylinositol-3-kinase (PI3K)/Akt signaling pathways in
colon tumorigenesis (40). In this study, high ALDH1B1 expression
in thyroid cancer was associated with poor OS in male patients, y
y
p g
We have shown the negative correlation between ALDH1A1/
B1 and several immune-stimulating genes. In addition, we
observed an opposite expression of the ALDH1A1/B1 and February 2022 | Volume 12 | Article 821958 Frontiers in Oncology | www.frontiersin.org 7 ALDH1A1/B1 in Thyroid Cancer Cui et al. A
B
C
FIGURE 5 | Correlation between ALDH1A1/A3/B1 expression and immune infiltration in thyroid cancer. (A–C) Correlation of ALDH1A1/A3/B1 expression with
infiltration levels of CD8+ T cell, CD4+ T cell, Treg cell, B cell, neutrophil, macrophage, dendritic cell, natural killer cell, and monocyte in thyroid cancer (total number of
patients with thyroid cancer, n = 223). Data were collected from the Tumor IMmune Estimation Resource. FIGURE 5 | Correlation between ALDH1A1/A3/B1 expression and immune infiltration in thyroid cancer. (A–C) Correlation of ALDH1A1/A3/B1 expression with
infiltration levels of CD8+ T cell, CD4+ T cell, Treg cell, B cell, neutrophil, macrophage, dendritic cell, natural killer cell, and monocyte in thyroid cancer (total number o
patients with thyroid cancer, n = 223). Data were collected from the Tumor IMmune Estimation Resource. MHC genes. During thyroid cancer progression, the immune
response relies on MHCs to recognize and present antigens to
cytotoxic T lymphocytes. Low levels of immune-stimulating
genes and MHCs would inhibit immune cell activation and induce a weaker response or promote immunological
unresponsiveness (44). Recently, several chemokines were
reported to be highly expressed in thyroid cancer tissues,
which might be involved in cancer progression and invasion FIGURE 6 | Interaction and Gene Ontology (GO)/pathway analysis of ALDH1A1/A3/B1. Protein–protein interaction network of ALDH1A1/A3/B1, constructed by the
online tool inBio Discover. FIGURE 6 | Interaction and Gene Ontology (GO)/pathway analysis of ALDH1A1/A3/B1. Protein–protein interaction network of ALDH1A1/A3/B1, constructed by the
online tool inBio Discover. February 2022 | Volume 12 | Article 821958 Frontiers in Oncology | www.frontiersin.org 8 ALDH1A1/B1 in Thyroid Cancer Cui et al. Cui et al. DISCUSSION A
B B A A B D C
D
xpression of ALDH1A1/B1 in thyroid cancer cell lines and thyroid cancer tissu C D D D C FIGURE 8 | Reduced expression of ALDH1A1/B1 in thyroid cancer cell lines and thyroid cancer tissues. Thyroid epithelial cell line nthy-ori 3-1 and thyroid cancer
cell lines (CHO-K1 and TPC-1) were collected, and the levels of ALDH1A1 (A) and ALDH1B1 (B) were detected by using qRT-PCR. Five thyroid cancer tissues and
corresponding adjacent tissues were collected from the First Hospital of Jilin University to verify the expression of ALDH1A1 (C) and ALDH1B1 (D) in patients. *P <
0.05; **P < 0.01; ****P < 0.0001. therapeutic targets for diagnosis and therapy. However, future
studies are required to validate our findings and promote the
clinical utility of ALDH1A1/B1 in thyroid cancer treatment. receptor has been implicated in non-small cell lung cancer (47),
gastric cancer (48), and breast cancer therapy (49). Of note, our
results showed a negative correlation between ALDH1A1/B1 and
several chemokines/receptors in thyroid cancer tissues. Thus,
some chemokines/receptors that showed decreased expression
in thyroid cancer tissues may be avoided as potential
therapeutic targets. DATA AVAILABILITY STATEMENT The datasets used and/or analyzed during the present study are
available from the corresponding author on reasonable request. In conclusion, we have revealed the aberrant expressions of
ALDH1A1/B1 in thyroid cancer tissues that were significantly
correlated with the OS of patients and with the pathological
stage. In addition, we found that elevated ALDH1A1/B1
expression was negatively associated with chemokine/receptor
expression and involved in several TILs. Thus, ALDH1A1/B1 are
potential biomarkers for thyroid cancer and might be valuable Frontiers in Oncology | www.frontiersin.org DISCUSSION ctivating Akt and Erk signaling pathway (45). Our results
carcinoma development (45). As soluble mediators secret
A
B
C
D
URE 7 | Correlation of ALDH1A1/A3/B1 with lymphocytes and immunomodulators. Relations between the abundance of immunostimulators (A), MHC (B),
mokines (C), and receptors (D) and ALDH1A1/A3/B1 expression and the top 2 genes displaying the greatest Spearman’s correlation with ALDH1A1/A3/B1
ession. Red and blue cells indicate positive and negative correlations, respectively. The color intensity is directly proportional to the strength of the correlatio
were collected from the TISIDB and TIMER database (total number of patients with thyroid cancer, n = 223). TILs, tumor-infiltrating lymphocytes, MHC ma
compatibility complex; TIMER, Tumor IMmune Estimation Resource. A
B A B C
D D C D FIGURE 7 | Correlation of ALDH1A1/A3/B1 with lymphocytes and immunomodulators. Relations between the abundance of immunostimulators (A), MHC (B),
chemokines (C), and receptors (D) and ALDH1A1/A3/B1 expression and the top 2 genes displaying the greatest Spearman’s correlation with ALDH1A1/A3/B1
expression. Red and blue cells indicate positive and negative correlations, respectively. The color intensity is directly proportional to the strength of the correlations. Data were collected from the TISIDB and TIMER database (total number of patients with thyroid cancer, n = 223). TILs, tumor-infiltrating lymphocytes, MHC major
histocompatibility complex; TIMER, Tumor IMmune Estimation Resource. carcinoma development (45). As soluble mediators secreted by
cells in tumor microenvironments, different chemokines recruit
various types of immune cells by binding to its corresponding
receptors (46). Moreover, targeting chemokine and chemokine by activating Akt and Erk signaling pathway (45). Our results
revealed the importance of chemokines and receptors in thyroid
cancer, which were in agreement with the study that emphasized
the crucial role of chemokine receptors during papillary thyroid February 2022 | Volume 12 | Article 821958 Frontiers in Oncology | www.frontiersin.org 9 Cui et al. Cui et al. ALDH1A1/B1 in Thyroid Cancer A
B
C
D
FIGURE 8 | Reduced expression of ALDH1A1/B1 in thyroid cancer cell lines and thyroid cancer tissues. Thyroid epithelial cell line nthy-ori 3-1 and thyroid cancer
cell lines (CHO-K1 and TPC-1) were collected, and the levels of ALDH1A1 (A) and ALDH1B1 (B) were detected by using qRT-PCR. Five thyroid cancer tissues and
corresponding adjacent tissues were collected from the First Hospital of Jilin University to verify the expression of ALDH1A1 (C) and ALDH1B1 (D) in patients. *P <
0.05; **P < 0.01; ****P < 0.0001. REFERENCES Front Immunol (2020) 11:784. doi: 10.3389/fimmu.2020.00784 7. Zhou C, Sun B. The Prognostic Role of the Cancer Stem Cell Marker Aldehyde
Dehydrogenase 1 in Head and Neck Squamous Cell Carcinomas: A Meta-Analysis. Oral Oncol (2014) 50(12):1144–8. doi: 10.1016/j.oraloncology.2014.08.018 24. Li B, Severson E, Pignon JC, Zhao H, Li T, Novak J, et al. Comprehensive
Analyses of Tumor Immunity: Implications for Cancer Immunotherapy. Genome Biol (2016) 17(1):174. doi: 10.1186/s13059-016-1028-7 8. Tomita H, Tanaka K, Tanaka T, Hara A. Aldehyde Dehydrogenase 1A1 in
Stem Cells and Cancer. Oncotarget (2016) 7(10):11018–32. doi: 10.18632/
oncotarget.6920 25. Wang W, Wang C, Xu H, Gao Y. Aldehyde Dehydrogenase, Liver Disease and
Cancer. Int J Biol Sci (2020) 16(6):921–34. doi: 10.7150/ijbs.42300 9. Kim D, Choi BH, Ryoo IG, Kwak MK. High NRF2 Level Mediates Cancer
Stem Cell-Like Properties of Aldehyde Dehydrogenase (ALDH)-High
Ovarian Cancer Cells: Inhibitory Role of All-Trans Retinoic Acid in
ALDH/NRF2 Signaling. Cell Death Disease (2018) 9(9):896. doi: 10.1038/
s41419-018-0903-4 26. Kurashige T, Shimamura M, Yasui K, Mitsutake N, Matsuse M, Nakashima M,
et al. Studies on Expression of Aldehyde Dehydrogenase in Normal and Cancerous
Tissues of Thyroids. Hormone Metab Res = Hormon- und Stoffwechselforschung =
Hormones metabolisme (2015) 47(3):194–9. doi: 10.1055/s-0034-1387770 10. Ruscito I, Darb-Esfahani S, Kulbe H, Bellati F, Zizzari IG, Rahimi Koshkaki H,
et al. The Prognostic Impact of Cancer Stem-Like Cell Biomarker Aldehyde
Dehydrogenase-1 (ALDH1) in Ovarian Cancer: A Meta-Analysis. Gynecologic
Oncol (2018) 150(1):151–7. doi: 10.1016/j.ygyno.2018.05.006 27. Nixon AB, Schalper KA, Jacobs I, Potluri S, Wang IM, Fleener C. Peripheral
Immune-Based Biomarkers in Cancer Immunotherapy: Can We Realize Their
Predictive Potential? J Immunotherapy Cancer (2019) 7(1):325. doi: 10.1186/
s40425-019-0799-2 11. Ginestier C, Hur MH, Charafe-Jauffret E, Monville F, Dutcher J, Brown M,
et al. ALDH1 is a Marker of Normal and Malignant Human Mammary Stem
Cells and a Predictor of Poor Clinical Outcome. Cell Stem Cell (2007) 1
(5):555–67. doi: 10.1016/j.stem.2007.08.014 28. Huang Y, Kim BYS, Chan CK, Hahn SM, Weissman IL, Jiang W. Improving
Immune-Vascular Crosstalk for Cancer Immunotherapy. Nat Rev Immunol
(2018) 18(3):195–203. doi: 10.1038/nri.2017.145 29. Axelrod ML, Cook RS, Johnson DB, Balko JM. Biological Consequences of
MHC-II Expression by Tumor Cells in Cancer. Clin Cancer Res an Off J Am
Assoc Cancer Res (2019) 25(8):2392–402. doi: 10.1158/1078-0432.ccr-18-3200 12. Lin CC, Lo MC, Moody RR, Stevers NO, Tinsley SL, Sun D. Doxycycline
Targets Aldehyde Dehydrogenase−Positive Breast Cancer Stem Cells. Oncol
Rep (2018) 39(6):3041–7. doi: 10.3892/or.2018.6337 30. REFERENCES 16. Chandrashekar DS, Bashel B, Balasubramanya SAH, Creighton CJ, Ponce-
Rodriguez I, Chakravarthi B, et al. UALCAN: A Portal for Facilitating Tumor
Subgroup Gene Expression and Survival Analyses. Neoplasia (New York NY)
(2017) 19(8):649–58. doi: 10.1016/j.neo.2017.05.002 1. Ancker OV, Krüger M, Wehland M, Infanger M, Grimm D. Multikinase
Inhibitor Treatment in Thyroid Cancer. Int J Mol Sci (2019) 21(1):10–29. doi: 10.3390/ijms21010010 17. Prevarskaya N, Skryma R, Shuba Y. Ion Channels in Cancer: Are Cancer
Hallmarks Oncochannelopathies? Physiol Rev (2018) 98(2):559–621. doi: 10.1152/physrev.00044.2016 2. Raue F, Frank-Raue K. Thyroid Cancer: Risk-Stratified Management and
Individualized Therapy. Clin Cancer Res an Off J Am Assoc Cancer Res (2016)
22(20):5012–21. doi: 10.1158/1078-0432.ccr-16-0484 18. Nagy Á , Munkácsy G, Győrffy B. Pancancer Survival Analysis of Cancer
Hallmark Genes. Sci Rep (2021) 11(1):6047. doi: 10.1038/s41598-021-84787-5 3. Varricchi G, Loffredo S, Marone G, Modestino L, Fallahi P, Ferrari SM, et al. The Immune Landscape of Thyroid Cancer in the Context of Immune
Checkpoint Inhibition. Int J Mol Sci (2019) 20(16):3934–65. doi: 10.3390/
ijms20163934 19. Li T, Fan J, Wang B, Traugh N, Chen Q, Liu JS, et al. TIMER: A Web Server
for Comprehensive Analysis of Tumor-Infiltrating Immune Cells. Cancer Res
(2017) 77(21):e108–10. doi: 10.1158/0008-5472.can-17-0307 20. Li T, Wernersson R, Hansen RB, Horn H, Mercer J, Slodkowicz G, et al. A
Scored Human Protein-Protein Interaction Network to Catalyze Genomic
Interpretation. Nat Methods (2017) 14(1):61–4. doi: 10.1038/nmeth.4083 4. Cabanillas ME, McFadden DG, Durante C. Thyroid Cancer. Lancet (London
England) (2016) 388(10061):2783–95. doi: 10.1016/s0140-6736(16)30172-6 5. Nilubol N, Zhang L, Kebebew E. Multivariate Analysis of the Relationship
Between Male Sex, Disease-Specific Survival, and Features of Tumor
Aggressiveness in Thyroid Cancer of Follicular Cell Origin. Thyroid Off J
Am Thyroid Assoc (2013) 23(6):695–702. doi: 10.1089/thy.2012.0269 21. Chefetz I, Grimley E, Yang K, Hong L, Vinogradova EV, Suciu R, et al. A Pan-
ALDH1A Inhibitor Induces Necroptosis in Ovarian Cancer Stem-Like Cells. Cell Rep (2019) 26(11):3061–3075.e6. doi: 10.1016/j.celrep.2019.02.032 22. Demir H, Dulgar O, Gulle BT, Turna H, Ilvan S. Prognostic Value of Aldehyde
Dehydrogenase 1 (ALDH1) in Invasive Breast Carcinomas. Bosnian J basic
Med Sci (2018) 18(4):313–9. doi: 10.17305/bjbms.2018.3094 6. Abdullah MI, Junit SM, Ng KL, Jayapalan JJ, Karikalan B, Hashim OH. Papillary Thyroid Cancer: Genetic Alterations and Molecular Biomarker
Investigations. Int J Med Sci (2019) 16(3):450–60. doi: 10.7150/ijms.29935 23. Petitprez F, Meylan M, de Reyniès A, Sautès-Fridman C, Fridman WH. The
Tumor Microenvironment in the Response to Immune Checkpoint Blockade
Therapies. FUNDING This work was supported by Scientific Research Funds of the
Education Department of Liaoning Province (JYTQN2020018). SUPPLEMENTARY MATERIAL YC performed the experiments. YC and YLiu analyzed the data
and wrote the article. LM collected samples. YC, YLiu, YLi, and
GW revised the article. GW conceived the idea and supervised
the project. All authors contributed to the article and approved
the submitted version. The Supplementary Material for this article can be found online at:
https://www.frontiersin.org/articles/10.3389/fonc.2022.821958/
full#supplementary-material. ETHICS STATEMENT All participants were given written informed consent, and the
current study has been approved by the ethics committee of the February 2022 | Volume 12 | Article 821958 10 ALDH1A1/B1 in Thyroid Cancer Cui et al. First Hospital of Jilin University. The patients/participants provided
their written informed consent to participate in this study. REFERENCES Fischer AK, Pham DL, Bösmüller H, Lengerke C, Wagner P, Bachmann C,
et al. Comprehensive in Situ Analysis of ALDH1 and SOX2 Reveals Increased
Expression of Stem Cell Markers in High-Grade Serous Carcinomas
Compared to Low-Grade Serous Carcinomas and Atypical Proliferative
Serous Tumors. Virchows Archiv an Int J Pathology (2019) 475(4):479–88. doi: 10.1007/s00428-019-02647-0 13. Li J, Zhang B, Yang YF, Jin J, Liu YH. Aldehyde Dehydrogenase 1 as a
Predictor of the Neoadjuvant Chemotherapy Response in Breast Cancer: A
Meta-Analysis. Medicine (2018) 97(34):e12056. doi: 10.1097/
md.0000000000012056 14. Wei D, Peng JJ, Gao H, Zhang T, Tan Y, Hu YH. ALDH1 Expression and the
Prognosis of Lung Cancer: A Systematic Review and Meta-Analysis. Heart
Lung Circulation (2015) 24(8):780–8. doi: 10.1016/j.hlc.2015.03.021 31. Ciccone V, Terzuoli E, Donnini S, Giachetti A, Morbidelli L, Ziche M. Stemness Marker ALDH1A1 Promotes Tumor Angiogenesis via Retinoic
Acid/HIF-1a/VEGF Signalling in MCF-7 Breast Cancer Cells. J Exp Clin
Cancer Res CR (2018) 37(1):311. doi: 10.1186/s13046-018-0975-0 15. Tang Z, Li C, Kang B, Gao G, Li C, Zhang Z. GEPIA: A Web Server for Cancer
and Normal Gene Expression Profiling and Interactive Analyses. Nucleic
Acids Res (2017) 45(W1):W98–102. doi: 10.1093/nar/gkx247 February 2022 | Volume 12 | Article 821958 Frontiers in Oncology | www.frontiersin.org 11 ALDH1A1/B1 in Thyroid Cancer Cui et al. Prostate Cancer Cell Line LNCaP. Exp Biol Med (Maywood NJ) (2007) 232
(6):762–71. doi: 10.3181/00379727-232-2320762 Prostate Cancer Cell Line LNCaP. Exp Biol Med (Maywood NJ) (2007) 232
(6):762–71. doi: 10.3181/00379727-232-2320762 32. Simões BM, O'Brien CS, Eyre R, Silva A, Yu L, Sarmiento-Castro A, et al. Anti-
Estrogen Resistance in Human Breast Tumors Is Driven by JAG1-NOTCH4-
Dependent Cancer Stem Cell Activity. Cell Rep (2015) 12(12):1968–77. doi: 10.1016/j.celrep.2015.08.050 43. Miftakhova R, Hedblom A, Semenas J, Robinson B, Simoulis A, Malm J, et al. Cyclin A1 and P450 Aromatase Promote Metastatic Homing and Growth of
Stem-Like Prostate Cancer Cells in the Bone Marrow. Cancer Res (2016) 76
(8):2453–64. doi: 10.1158/0008-5472.can-15-2340 33. Panigoro SS, Kurnia D, Kurnia A, Haryono SJ, Albar ZA. ALDH1 Cancer
Stem Cell Marker as a Prognostic Factor in Triple-Negative Breast Cancer. Int
J Surg Oncol (2020) 2020:7863243. doi: 10.1155/2020/7863243 44. Rock KL, Reits E, Neefjes J. Present Yourself! By MHC Class I and MHC Class II
Molecules. Trends Immunol (2016) 37(11):724–37. doi: 10.1016/j.it.2016.08.010 34. Chen MF, Chen PT, Lu MS, Chen WC. Role of ALDH1 in the Prognosis of
Esophageal Cancer and its Relationship With Tumor Microenvironment. REFERENCES Mol
Carcinogenesis. (2018) 57(1):78–88. doi: 10.1002/mc.22733 45. Zhu X, Bai Q, Lu Y, Lu Y, Zhu L, Zhou X, et al. Expression and Function of
CXCL12/CXCR4/CXCR7 in Thyroid Cancer. Int J Oncol (2016) 48(6):2321–9. doi: 10.3892/ijo.2016.3485 35. Wang J, Wang L, Ho CT, Zhang K, Liu Q, Zhao H. Garcinol From Garcinia
Indica Downregulates Cancer Stem-Like Cell Biomarker ALDH1A1 in
Nonsmall Cell Lung Cancer A549 Cells Through DDIT3 Activation. J Agric
Food Chem (2017) 65(18):3675–83. doi: 10.1021/acs.jafc.7b00346 46. Mollica Poeta V, Massara M, Capucetti A, Bonecchi R. Chemokines and
Chemokine Receptors: New Targets for Cancer Immunotherapy. Front
Immunol (2019) 10:379. doi: 10.3389/fimmu.2019.00379 47. Wang CL, Sun BS, Tang Y, Zhuang HQ, Cao WZ. CCR1 Knockdown
Suppresses Human Non-Small Cell Lung Cancer Cell Invasion. J Cancer
Res Clin Oncol (2009) 135(5):695–701. doi: 10.1007/s00432-008-0505-0 36. Nie S, Qian X, Shi M, Li H, Peng C, Ding X, et al. ALDH1A3 Accelerates
Pancreatic Cancer Metastasis by Promoting Glucose Metabolism. Front Oncol
(2020) 10:915. doi: 10.3389/fonc.2020.00915 37. Lee S, Bae JS, Jung CK, Chung WY. Extensive Lymphatic Spread of Papillary
Thyroid Microcarcinoma is Associated With an Increase in Expression of
Genes Involved in Epithelial-Mesenchymal Transition and Cancer Stem Cell-
Like Properties. Cancer Med (2019) 8(15):6528–37. doi: 10.1002/cam4.2544 48. Aldinucci D, Casagrande N. Inhibition of the CCL5/CCR5 Axis Against the
Progression of Gastric Cancer. Int J Mol Sci (2018) 19(5):1477–93. doi: 10.3390/ijms19051477 49. Bonapace L, Coissieux MM, Wyckoff J, Mertz KD, Varga Z, Junt T, et al. Cessation
of CCL2 Inhibition Accelerates Breast Cancer Metastasis by Promoting
Angiogenesis. Nature (2014) 515(7525):130–3. doi: 10.1038/nature13862 49. Bonapace L, Coissieux MM, Wyckoff J, Mertz KD, Varga Z, Junt T, et al. Cessation
of CCL2 Inhibition Accelerates Breast Cancer Metastasis by Promoting
Angiogenesis. Nature (2014) 515(7525):130–3. doi: 10.1038/nature13862 38. Shimamura M, Kurashige T, Mitsutake N, Nagayama Y. Aldehyde
Dehydrogenase Activity Plays No Functional Role in Stem Cell-Like
Properties in Anaplastic Thyroid Cancer Cell Lines. Endocrine (2017) 55
(3):934–43. doi: 10.1007/s12020-016-1224-y Conflict of Interest: The authors declare that the research was conducted in the
absence of any commercial or financial relationships that could be construed as a
potential conflict of interest. 39. Mameishvili E, Serafimidis I, Iwaszkiewicz S, Lesche M, Reinhardt S, Bölicke N,
et al. Aldh1b1 Expression Defines Progenitor Cells in the Adult Pancreas and is
Required for Kras-Induced Pancreatic Cancer. Proc Natl Acad Sci USA (2019) 116
(41):20679–88. February 2022 | Volume 12 | Article 821958 Frontiers in Oncology | www.frontiersin.org REFERENCES doi: 10.1073/pnas.1901075116 Publisher’s Note: All claims expressed in this article are solely those of the authors
and do not necessarily represent those of their affiliated organizations, or those of
the publisher, the editors and the reviewers. Any product that may be evaluated in
this article, or claim that may be made by its manufacturer, is not guaranteed or
endorsed by the publisher. 40. Singh S, Arcaroli J, Chen Y, Thompson DC, Messersmith W, Jimeno A, et al. ALDH1B1 Is Crucial for Colon Tumorigenesis by Modulating Wnt/b-
Catenin, Notch and PI3K/Akt Signaling Pathways. PloS One (2015) 10(5):
e0121648. doi: 10.1371/journal.pone.0121648 41. Gangavarapu KJ, Azabdaftari G, Morrison CD, Miller A, Foster BA, Huss WJ. Aldehyde Dehydrogenase and ATP Binding Cassette Transporter G2
(ABCG2) Functional Assays Isolate Different Populations of Prostate Stem
Cells Where ABCG2 Function Selects for Cells With Increased Stem Cell
Activity. Stem Cell Res Ther (2013) 4(5):132. doi: 10.1186/scrt343 Copyright © 2022 Cui, Liu, Mu, Li and Wu. This is an open-access article distributed
under the terms of the Creative Commons Attribution License (CC BY). The
use, distribution or reproduction in other forums is permitted, provided the original
author(s) and the copyright owner(s) are credited and that the original publication in
this journal is cited, in accordance with accepted academic practice. No use,
distribution or reproduction is permitted which does not comply with these terms. 42. Trasino SE, Harrison EH, Wang TT. Androgen Regulation of Aldehyde
Dehydrogenase 1A3 (ALDH1A3) in the Androgen-Responsive Human February 2022 | Volume 12 | Article 821958 Frontiers in Oncology | www.frontiersin.org 12
|
https://openalex.org/W2903672731
|
https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0208636&type=printable
|
English
| null |
Epitope specificity of anti-synapsin autoantibodies: Differential targeting of synapsin I domains
|
PloS one
| 2,018
|
cc-by
| 8,006
|
RESEARCH ARTICLE Objective To identify the specific domains of the presynaptic protein synapsin targeted by recently
described autoantibodies to synapsin. Editor: Edgar Meinl, Ludwig-Maximilians-
Universitat Munchen, GERMANY
Received: July 4, 2018
Accepted: November 20, 2018
Published: December 13, 2018
Copyright: © 2018 Mertens et al. This is an open
access article distributed under the terms of the
Creative Commons Attribution License, which
permits unrestricted use, distribution, and
reproduction in any medium, provided the original
author and source are credited. Editor: Edgar Meinl, Ludwig-Maximilians-
Universitat Munchen, GERMANY
Received: July 4, 2018
Accepted: November 20, 2018
Published: December 13, 2018 Abstract Citation: Mertens R, Melchert S, Gitler D, Schou
MB, Saether SG, Vaaler A, et al. (2018) Epitope
specificity of anti-synapsin autoantibodies:
Differential targeting of synapsin I domains. PLoS
ONE 13(12): e0208636. https://doi.org/10.1371/
journal.pone.0208636 Epitope specificity of anti-synapsin
autoantibodies: Differential targeting of
synapsin I domains Robert Mertens1☯, Sarah Melchert1☯, Daniel Gitler2, Morten Brix Schou3,4, Sverre
Georg Saether3,4, Arne Vaaler3,4, Johannes Piepgras1, Elena Kochova1, Fabio Benfenati5,
Gudrun Ahnert-Hilger1, Klemens Ruprecht6, Markus Ho¨ltjeID1* 1 Institute of Integrative Neuroanatomy, Charite´ –Universita¨tsmedizin Berlin, corporate member of Freie
Universita¨t Berlin, Humboldt-Universita¨t zu Berlin, and Berlin Institute of Health, Berlin, Germany,
2 Department of Physiology and Cell Biology, Faculty of Health Sciences and Zlotowski Center for
Neuroscience, Ben-Gurion University of the Negev, Beer-Sheva, Israel, 3 Department of Psychiatry,
St. Olav’s University Hospital, Trondheim, Norway, 4 Department of Mental Health, Norwegian University of
Science and Technology, Trondheim, Norway, 5 Center for Synaptic Neuroscience and Technology, Istituto
Italiano di Tecnologia, Genova, Italy, 6 Department of Neurology, Charite´ –Universita¨tsmedizin Berlin,
corporate member of Freie Universita¨t Berlin, Humboldt-Universita¨t zu Berlin, and Berlin Institute of Health,
Berlin, Germany ☯These authors contributed equally to this work. Methods Sera of 20 and CSF of two patients with different psychiatric and neurological disorders pre-
viously tested positive for immunoglobulin (Ig)G antibodies to full-length synapsin were
screened for IgG against synapsin I domains using HEK293 cells transfected with con-
structs encoding different domains of rat synapsin Ia. Additionally, IgG subclasses were
determined using full-length synapsin Ia. Serum and CSF from one patient were also
screened for IgA autoantibodies to synapsin I domains. Sera from nine and CSF from two
healthy subjects were analyzed as controls. Copyright: © 2018 Mertens et al. This is an open
access article distributed under the terms of the
Creative Commons Attribution License, which
permits unrestricted use, distribution, and
reproduction in any medium, provided the original
author and source are credited. ☯These authors contributed equally to this work.
* markus.hoeltje@charite.de Data Availability Statement: All relevant data are
within the manuscript. Data Availability Statement: All relevant data are
within the manuscript. IgG in serum from 12 of 20 IgG synapsin full-length positive patients, but from none of the
healthy controls, bound to synapsin domains. Of these 12 sera, six bound to the A domain,
five to the D domain, and one to the B- (and possibly A-), D-, and E-domains of synapsin I. IgG antibodies to the D-domain were also detected in one of the CSF samples. Determina-
tion of IgG subclasses detected IgG1 in two sera and one CSF, IgG2 in none of the samples,
IgG3 in two sera, and IgG4 in eight sera. One patient known to be positive for IgA antibodies
to full-length synapsin had IgA antibodies to the D-domain in serum and CSF. Funding: DG was supported by Israel Science
Foundation (ISF) grant 1427/12.KR was supported
by the German Ministry of Education and Research
(BMBF/KKNMS, Competence Network Multiple
Sclerosis). All other authors report no disclosures. Competing interests: The authors have declared
that no competing interests exist. 1 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0208636
December 13, 2018 Autoantibodies and synapsin domains Conclusions Anti-synapsin autoantibodies preferentially bind to either the A- or the D-domain of synapsin I. Ethical approval The study was approved by the institutional review board of Charite´– Universita¨tsmedizin
Berlin (EA1/182/10; EA1/129/14, EA1/096/12) and the regional committee for medical and
health research ethics, central Norway (2011/137). All participants provided written informed
consent. Introduction By using a combination of immunoprecipitation methods, mass spectrometry, immunohis-
tochemistry of brain sections from wild type and synapsin-knock mice, as well as cell-based
assays and biochemistry we recently identified the presynaptic vesicle-associated protein
synapsin as a novel target of antineuronal autoantibodies in serum and CSF samples from a
patient with limbic encephalitis [1]. In a follow-up study using rat and human synapsin I trans-
fected HEK293 cells as well as biochemical analyses, serum IgG to full-length rat synapsin Ia
could be detected in 23 of 375 (6.1%) patients with different neurological and psychiatric dis-
eases, including patients with psychotic, bipolar or depressive disorders and multiple sclerosis
(MS) or clinically isolated syndrome (CIS), but not in 97 healthy controls [2]. Concomitant
serum autoantibodies, mostly low titer anti NMDAR antibodies, were detected in eight of the
patients. Synapsin I and II are major synaptic vesicle proteins of the adult brain, regulating the avail-
ability of synaptic vesicles and thus affecting synaptic transmission [3; 4; 5]. Synapsins are
composed of different highly preserved domains that were defined by sequence homology
across species [6]. Synapsin Ia and Ib are two splice variants, which share the A-, B-, C- and D-
domains, but differ in the C-terminal region, consisting of the E domain in synapsin Ia and
the shorter F-domain in synapsin Ib (see also Fig 1A). Distinct functional roles, such as inter-
ference with the generation of excitatory postsynaptic potentials attributed to domain A of
synapsin I, were ascribed to individual domains of synapsin I [7]. Membrane insertion
involves domain C of synapsin I [8] and interaction with SH3 domain-containing endocytic
proteins was shown for the D-domain of synapsin I [9]. The pathophysiological relevance of antibodies to synapsin remains to be clarified. Our
finding that antibodies to synapsin are associated with different clinical phenotypes raises the
question whether this may be related to distinct functions of synapsin and to the recognition
of different domains of synapsin by autoantibodies to synapsin. To better understand the
potential pathophysiological relevance of antibodies to synapsin, we here characterized in
molecular detail the specific domains of synapsin targeted by synapsin autoantibodies. Patients Patient sera investigated in this study were previously shown to contain antibodies to full-
length rat synapsin Ia and included sera from 16 patients positive for IgG autoantibodies to rat
synapsin Ia identified from a total of 207 patients admitted to acute psychiatric inpatient care
at the Department of Psychiatry, St. Olav’s University Hospital, Trondheim, Norway, between PLOS ONE | https://doi.org/10.1371/journal.pone.0208636
December 13, 2018 2 / 14 Autoantibodies and synapsin domains Fig 1. Specific recognition of synapsin I domains expressed in transfected HEK cells by domain-specific antibodies. (A) Domain structure of the synapsin
isoforms Ia and Ib. The N-terminal domains A-C are highly conserved in all synapsins. The C-terminal region is variable due to heterogeneous combinations of
domains. (B) Diagram of the synapsin Ia fragments examined to investigate targeting of patient synapsin autoantibodies. (C-H) HEK293 cells were transfected
with rat synapsin Ia constructs, comprising the eYFP-tagged fragment ABC (C), eGFP-tagged BC (D), eGFP-tagged C (E), eGFP-tagged DE (F), eGFP-tagged D
(G), or eGFP-tagged E domain (H). Cells were fixed, permeabilized and incubated with antibodies either directed against the A-domain of synapsin I and II (rabbit
anti-synapsin A; dilution 1:500), the D-domain of synapsin I (mouse anti-synapsin D; dilution 1:500), the C-domain of synapsin I/II (rabbit anti-synapsin C;
dilution 1:500), or the E-domain of synapsin Ia (rabbit anti-synapsin E; dilution 1:200). Antibody binding was visualized using Alexa Fluor 549-coupled secondary
antibodies. Transfected cells were correctly recognized by the respective antibodies that colocalized with the signal of the tags, confirming domain identities and
detectability of the various domains by anti-synapsin antibodies. htt
//d i
/10 1371/j
l
0208636 001 Fig 1. Specific recognition of synapsin I domains expressed in transfected HEK cells by domain-specific antibodies. (A) Domain structure of the synapsin
isoforms Ia and Ib. The N-terminal domains A-C are highly conserved in all synapsins. The C-terminal region is variable due to heterogeneous combinations of
domains. (B) Diagram of the synapsin Ia fragments examined to investigate targeting of patient synapsin autoantibodies. (C-H) HEK293 cells were transfected
with rat synapsin Ia constructs, comprising the eYFP-tagged fragment ABC (C), eGFP-tagged BC (D), eGFP-tagged C (E), eGFP-tagged DE (F), eGFP-tagged D
(G), or eGFP-tagged E domain (H). PLOS ONE | https://doi.org/10.1371/journal.pone.0208636
December 13, 2018 Secondary antibodies Mouse antibodies were visualized using Alexa Fluor 594 goat anti-mouse IgG (MoBiTec, Go¨t-
tingen, Germany; # A11032). An Alexa Fluor 488 goat anti-guinea pig IgG (#A11073) was also
from MoBiTec. Alexa Fluor 594 goat anti-rabbit IgG (#A11037; Life Technologies, Carlsbad,
CA, USA) was applied to detect bound rabbit antibodies. Bound human antibodies of the
immunoglobulin type G (total) were visualized by incubation with Alexa Fluor 594 labeled
goat anti-human IgG directed against the Fc fragment (Dianova GmbH, Hamburg, Germany;
#109-585-008). Visualization of human IgG subclass-specific antibodies was achieved using
Alexa Fluor 647 labeled goat anti-human IgG1, IgG2, IgG3 and IgG4 antibodies (Southern Bio-
tech, Birmingham, USA; #9052–31, #9070–31, #9210–31, #9200–31). Detection of immuno-
globulin A antibodies in the patients’ CSF or serum was performed by using a Texas Red 594
coupled goat anti-human IgA secondary antibody (Dianova; #109-076-011). Cell-based assays for synapsin domains and full-length synapsin Ia Human embryonic kidney (HEK) 293 cells were cultured in 24-well multiplates to 70% con-
fluency and transfected with 0.5–1 μg plasmid cDNA per well for 24 hours using Lipofecta-
mine (Gibco/BRL Life Technologies, Eggenstein, Germany). Transfected cells were fixed with
ice-cold 80% methanol for 20 minutes at -20˚C (or 4% paraformaldehyde at 4˚C for detection
of IgA immunoreactivity). Cells were subsequently permeabilized with 0.1% Triton X-100. Thereafter, cells were incubated with either the domain specific primary antibodies at the indi-
cated concentrations or with patient or control serum or CSF at a dilution of 1:320. Patients Cells were fixed, permeabilized and incubated with antibodies either directed against the A-domain of synapsin I and II (rabbit
anti-synapsin A; dilution 1:500), the D-domain of synapsin I (mouse anti-synapsin D; dilution 1:500), the C-domain of synapsin I/II (rabbit anti-synapsin C;
dilution 1:500), or the E-domain of synapsin Ia (rabbit anti-synapsin E; dilution 1:200). Antibody binding was visualized using Alexa Fluor 549-coupled secondary
antibodies. Transfected cells were correctly recognized by the respective antibodies that colocalized with the signal of the tags, confirming domain identities and
detectability of the various domains by anti-synapsin antibodies. https://doi org/10 1371/journal pone 0208636 g001 September 2011 and March 2012 [10]. Additionally, three sera and 1 CSF from three anti-
synapsin IgG positive patients with a clinically isolated syndrome (CIS) were available from a
cohort total of 78 patients with a CIS or early relapsing-remitting MS (RRMS) collected at the
NeuroCure Clinical Research Center/Department of Neurology, Charite´ –Universita¨tsmedizin
Berlin. The patients’ demographic and clinical findings were recently described in detail [2]. In
addition, we analyzed serum and cerebrospinal fluid (CSF) from the 69-year old male index
patient diagnosed with limbic encephalitis in 2013 in whom IgA and IgG autoantibodies to
synapsin I and II were originally identified [1]. As controls, nine sera and two CSF samples
from healthy subjects were obtained at the NeuroCure Clinical Research Center/Department
of Neurology, Charite´ –Universita¨tsmedizin Berlin. 3 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0208636
December 13, 2018 Autoantibodies and synapsin domains Primary antibodies A monoclonal mouse antibody against the D-domain of synapsin I (#106011) and a polyclonal
rabbit antibody against the A-domain of synapsin I/II (#106002) were purchased from Synap-
tic Systems (Go¨ttingen, Germany). Rabbit polyclonal antibodies against the C-domain of
synapsin I/II and the E-domain of synapsin Ia were developed by one of us (F.B.), as described
earlier [11]. A guinea pig polyclonal antibody against GFP used in some experiments to
enhance endogenous fluorescence was also obtained from Synaptic Systems (#132005). Synapsin plasmids The expression constructs for the various synapsin domains have been described earlier [6]. In
brief, domain cDNAs were cloned into either pEGFP or pEYFP C1-C3 vectors. The following
domains or domain combinations constituting sequence fragments of rat synapsin Ia (Uni-
Prot/SWISS-PROT AAA42145.1) were used in our study: pEYFP-ABC; pEGFP-BC; pEGFP-C;
pEGFP-D; pEGFP-DE and pEGFP-E. From the sequence of rat synapsin Ia, the following frag-
ments were constructed: these constructs contained the N-terminal ABC fragment (aa 1–416),
the BC fragment (aa 31–416), the C-domain (aa 113–420), the DE fragment (aa 415–704), the
D-domain (aa 414–651) and the E-domain (aa 655–704) (for detail see also Fig 1A, B). Rat full-
length synapsin Ia cDNA was tagged on the N-terminus with enhanced green fluorescent pro-
tein (eGFP) based on a pEGFP-C1 backbone. Cell-based assays for detection of antibodies to synapsin I domains From the sequence of rat synapsin Ia, six different combinatory domain fragments or single
domains were constructed (Fig 1A and 1B). A rationale to use fragments from rat synapsin
sequence was the fact that these constructs have been extensively characterized for targeting
and binding to presynaptic terminals [6]. Of note, human and rat synapsin I share a 95% iden-
tity of the primary structure [13] and binding of patient antibodies to rat and human protein
exhibited a very similar pattern [2]. To establish cell-based assays for the detection of antibodies to synapsin I domains, we
transfected six different rat synapsin Ia constructs (Fig 1B) in human embryonic kidney
(HEK) 293 cells. Subsequently, transfected cells were incubated with antibodies specifically
recognizing the A-, C-, D- or E-domain of synapsin I. As shown in Fig 1C–1H all six synapsin I fragments were clearly detectable by their endoge-
nous fluorescence tags (eYFP for the ABC-fragment and eGFP for all other constructs). Fur-
thermore, all individual or combined synapsin Ia domains were specifically recognized by
antibodies directed against the respective domains indicating proper assay function. Antibody index calculation HEK 293 cells transfected with rat synapsin Ia were processed as described above and incu-
bated with increasing dilutions (1:320, 1:1000, 1:3200, 1:10.000, and 1:32,000) of patient CSF
or serum to determine titers of anti-synapsin IgG in a paired CSF/serum sample. Alexa Fluor
594 goat anti-human IgG served as secondary antibody. Antibody index (AI) for IgG PLOS ONE | https://doi.org/10.1371/journal.pone.0208636
December 13, 2018 4 / 14 Autoantibodies and synapsin domains antibodies to synapsin Ia was calculated as previously described by the formula: AI = (anti-
synapsin titer CSF-1/anti-synapsin titer serum-1)/(total Ig CSF/total Ig serum). AI values above
four were considered as evidence of an intrathecal antibody synthesis [12]. Autoantibodies and synapsin domains Table 1. Demographic, antibody specificity and clinical findings of the 13 patients with serum or CSF antibodies to synapsin Ia fragments identified in this study. Antibodies to synapsin predominantly target the A- or D-domain of
synapsin Ia To decipher the domain specificity of autoantibodies to synapsin Ia, we analyzed a total of 20
serum and two CSF samples, which contained antibodies to full-length rat synapsin Ia as iden-
tified in our previous studies [1; 2]. For control, nine sera and two CSF samples from healthy
controls were analyzed as well. We first screened sera and one CSF from 19 patients positive for rat synapsin IgG for their
reactivity with the various synapsin domains. Sixteen of these patients were diagnosed with
psychiatric disorders and three patients with a CIS. CSF was available from one of the patients
with a CIS. A summary of the findings is presented in Table 1. Out of the sera of 16 patients with psychiatric disorders positive for antibodies to full-length
synapsin, 11 sera exhibited a detectable immunoreactivity against one or more of the synapsin
Ia domains. Out of the three patients diagnosed with a CIS, serum and CSF from one patient
showed immunoreactivity to synapsin Ia domains. Among these 12 positive sera, seven were
positive for the ABC fragment (58,3%). Among these seven sera, only one serum was also posi-
tive for the BC fragment and none was positive for the single C-domain. Thus, sera of six
patients were likely exclusively positive for the A-domain (as exemplarily shown for one
patient in Fig 2A) and one proved to be positive for both the A- and B-domains, or solely the
B-domain (PSY_0149, the only patient with multiple targeted domains, Table 1). A direct
approach towards testing reactivity to the A- or B-domain alone was not possible due to the
lack of individual constructs for these domains. The only serum that reacted with both the
ABC and the BC fragment also reacted with the C-terminal domains D and E. The five sera (41,7%) that did not react with the ABC fragment exhibited reactivity to either
the DE fragment and the D-domain (three), the DE fragment alone (one) or the D-domain
alone (one). None of these five sera showed reactivity towards the E-domain alone, indicating
that the D-domain was the targeted antigenic region of synapsin Ia. A CSF sample available 5 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0208636
December 13, 2018 Serum code
Höltje et al.,
2017
Sex
Age
(years)
ABC
BC
C
DE
D
E
Synapsin Ia
full length
titer
Diagnoses
Index patient
serum
m
69
-
-
-
+
(IgA)
+
(IgA)
-
1:1000–
1:3200
Limbic encephalitis
Index patient
CSF
„
„
-
-
-
+
(IgA)
+
(IgA)
-
1:1000–
1:3200
„
CIS_0036
f
34
-
-
-
-
-
-
1:3200
Clinically isolated syndrome; panic/anxiety and recurrent depressive disorder
CIS_0064
f
51
-
-
-
-
-
-
1:1000
Clinically isolated syndrome
CIS_0117
serum,
f
24
-
-
-
+
+
-
1:10000
Clinically isolated syndrome
CIS_0117
CSF
„
„
-
-
-
+
+
-
1:1000
„
PSY_0012
f
29
+
-
-
-
-
-
1:320
Bipolar affective disorder type 1, mixed episode
PSY_0032
f
20
+
-
-
-
-
-
1:320
Bipolar affective disorder type 2, depressive episode
PSY_0045
f
32
-
-
-
-
-
-
1:1000
Persistent personality change after catastrophic experience
PSY_0068
f
52
-
-
-
+
-
-
1:10000
Recurrent depressive disorder, severe episode; coma for 12 days at age 8 years
after accident
PSY_0082
m
26
-
-
-
-
-
-
1:320
Acute polymorphic psychotic disorder; Mo¨bius syndrome
PSY_0094
m
39
-
-
-
-
+
-
1:100000
Alcohol abuse, ADHD, bipolar affective disorder type 2, emotionally unstable
personality disorder
PSY_0141
f
21
-
-
-
+
+
-
1:1000
Alcohol dependency syndrome, ADHD, PTSD, mixed obsessional thoughts and
acts
PSY_0149
m
51
+
+
-
+
+
+
1:100000
Bipolar affective disorder type 1, manic episode; alcohol dependency syndrome;
hepatitis C
PSY_0171
f
22
-
-
-
+
+
-
1:32000
Moderate depressive episode; SLE
PSY_0183
f
31
-
-
-
-
-
-
1:3200
Recurrent depressive disorder, moderate episode; personality disorder
PSY_1851
f
58
+
-
-
-
-
-
1:1000
Bipolar disorder type 1, manic episode with psychotic symptoms; MS; later
diagnosed organic affective disorder
PSY_1852
m
68
+
-
-
-
-
-
1:320
Bipolar affective disorder type 1, manic episode; paranoid psychosis
PSY_0189
f
77
+
-
-
-
-
-
1:3200
Recurrent depressive disorder
PSY_0201
f
44
-
-
-
-
-
-
1:1000
Recurrent depressive disorder, moderate episode; alcohol dependency syndrome;
epileptic seizures as child; probably anoxic injury during birth
PSY_0203
m
27
+
-
-
-
-
-
1:320
Paranoid schizophrenia; multiple drug abuse
PSY_0204
m
60
-
-
-
-
-
-
1:1000
Recurrent depressive disorder, mild episode; cannabinoid abuse; melanoma;
hepatitis C; psoriasis from a patient whose serum reacted with the DE fragment and the D-domain likewise reacted
with the DE fragment and the D-domain. Cells were incubated with patient serum or CSF at a dilu
1:320. Bound IgG was detected using an Alexa 594-coupled secondary antibody to the Fc fragment of human
the patient’s serum and CSF demonstrated positive immunoreactivity that co-localized with the eGFP tag of s
DE and the single D-domain, but with none of the other constructs. (C) Serum and CSF of two healthy contr
(control IgG) did not bind to cells transfected with synapsin ABC or DE. https://doi.org/10.1371/journal.pone.0208636.g002 Fig 2. Serum IgG autoantibodies against the A-domain of a patient with bipolar affective disorder and serum and
CSF IgG autoantibodies against the D-domain of a patient with a clinically isolated syndrome (CIS). (A) HEK293
cells were transfected with eYFP-tagged synapsin I fragment ABC, eGFP-tagged DE, BC or C. Cells were incubated
with patient serum at a dilution of 1:320. Bound IgG was detected using an Alexa 594-coupled secondary antibody to
the Fc fragment of human IgG. The patient’s serum demonstrated positive immunoreactivity that co-localized with the
eYFP tag of synapsin ABC, but with none of the other constructs. (B) HEK293 cells were transfected with eYFP-tagged
synapsin I fragment ABC, eGFP-tagged DE, D or E. Cells were incubated with patient serum or CSF at a dilution of
1:320. Bound IgG was detected using an Alexa 594-coupled secondary antibody to the Fc fragment of human IgG. Both
the patient’s serum and CSF demonstrated positive immunoreactivity that co-localized with the eGFP tag of synapsin
DE and the single D-domain, but with none of the other constructs. (C) Serum and CSF of two healthy controls
(control IgG) did not bind to cells transfected with synapsin ABC or DE. https://doi org/10 1371/journal pone 0208636 g002 Fig 2. Serum IgG autoantibodies against the A-domain of a patient with bipolar affective disorder and serum and
CSF IgG autoantibodies against the D-domain of a patient with a clinically isolated syndrome (CIS). (A) HEK293
cells were transfected with eYFP-tagged synapsin I fragment ABC, eGFP-tagged DE, BC or C. Cells were incubated
with patient serum at a dilution of 1:320. Bound IgG was detected using an Alexa 594-coupled secondary antibody to
the Fc fragment of human IgG. The patient’s serum demonstrated positive immunoreactivity that co-localized with the
eYFP tag of synapsin ABC, but with none of the other constructs. (B) HEK293 cells were transfected with eYFP-tagged
synapsin I fragment ABC, eGFP-tagged DE, D or E. The results of the cell-based assays for this patient
with a CIS are shown in Fig 2B. For this patient, antibody titers of the serum/CSF pair were
determined using full-length synapsin Ia and the antibody index (AI) was calculated. The AI
value of 19,6 clearly indicated an intrathecal synthesis of IgG to synapsin I. Additionally, all
nine control sera and two CSF samples applied at the same dilution as the patients’ sera were
negative for IgG synapsin autoantibodies, as exemplarily shown for two healthy subjects in Fig
2C. p
p
Presence (+) or absence (-) of immunoreactivity is indicated. Diagnoses, antibody titers to full length synapsin and concomitant autoantibodies (not shown) have been published previously [2]. ADHD = attention deficit hyperactivity disorder; f = female; m = male; MS = multiple sclerosis; PTSD = post traumatic stress disorder; SLE = systemic lupus
erytematodus
https://doi.org/10.1371/journal.pone.0208636.t001
PLOS ONE | https://doi.org/10.1371/journal.pone.0208636
December 13, 2018
6 / 14 from a patient whose serum reacted with the DE fragment and the D-domain likewise reacted
with the DE fragment and the D-domain. The results of the cell-based assays for this patient
with a CIS are shown in Fig 2B. For this patient, antibody titers of the serum/CSF pair were
determined using full-length synapsin Ia and the antibody index (AI) was calculated. The AI
value of 19,6 clearly indicated an intrathecal synthesis of IgG to synapsin I. Additionally, all
nine control sera and two CSF samples applied at the same dilution as the patients’ sera were
negative for IgG synapsin autoantibodies, as exemplarily shown for two healthy subjects in Fig
2C. 6 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0208636
December 13, 2018 PLOS ONE | https://doi.org/10.1371/journal.pone.0208636
December 13, 2018 Autoantibodies and synapsin domains Fig 2. Serum IgG autoantibodies against the A-domain of a patient with bipolar affective disorder and se
CSF IgG autoantibodies against the D-domain of a patient with a clinically isolated syndrome (CIS). (A)
cells were transfected with eYFP-tagged synapsin I fragment ABC, eGFP-tagged DE, BC or C. Cells were incu
with patient serum at a dilution of 1:320. Bound IgG was detected using an Alexa 594-coupled secondary anti
the Fc fragment of human IgG. The patient’s serum demonstrated positive immunoreactivity that co-localize
eYFP tag of synapsin ABC, but with none of the other constructs. (B) HEK293 cells were transfected with eYF
synapsin I fragment ABC, eGFP-tagged DE, D or E. Cells were incubated with patient serum or CSF at a dilution of
1:320. Bound IgG was detected using an Alexa 594-coupled secondary antibody to the Fc fragment of human IgG. Both
the patient’s serum and CSF demonstrated positive immunoreactivity that co-localized with the eGFP tag of synapsin
DE and the single D-domain, but with none of the other constructs. (C) Serum and CSF of two healthy controls
(control IgG) did not bind to cells transfected with synapsin ABC or DE. https://doi.org/10.1371/journal.pone.0208636.g002 7 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0208636
December 13, 2018 Autoantibodies and synapsin domains Of note, all patient and control samples were also tested for IgA reactivity to the ABC and
DE fragments. All samples remained negative, thereby confirming our previous results
obtained for full-length synapsin [2]. As a further control, 10 patient sera from that study nega-
tive for antibodies to full-length synapsin were tested for IgG and IgA reactivity to the ABC or
DE fragment. Also in this case no reactivity to one of the fragments was detected. IgA antibodies from the index patient with limbic encephalitis target the
D-domain of synapsin Ia We also analyzed serum and CSF from our index patient, who had high titers of anti-synapsin
IgA and IgG in serum and CSF [1]. Both serum and CSF IgA clearly reacted with the DE frag-
ment and the D-domain, but not with the ABC or E construct (Fig 3A). Of note, we could not
detect reactivity of IgG antibodies to any of the domains despite the fact that serum and CSF
IgG reacted with full-length synapsin. Serum and CSF IgA from two healthy subjects did not
react with any of the synapsin constructs (Fig 3B). After successful determination of domain specificities we additionally aimed to identify the
respective IgG subclasses involved in autoimmunity against synapsin. To this end, we trans-
fected HEK 293 cells with full-length synapsin Ia, again incubated fixed cells with patient sera
that reacted with any of the fragments as well as with the CSF of CIS117 at a dilution of 1:320
and applied secondary antibodies specific for the four subclasses IgG1, IgG2, IgG3 and IgG4
(Fig 4A–4C). As a result, 1 serum and the corresponding CSF exclusively contained IgG1, 2
sera exclusively showed IgG3 reactivity, 7 sera only showed IgG4 reactivity, and one serum was
positive for IgG1 and IgG4. No IgG2 reactivity was observed (for summary, see Table 2). Note-
worthy, the patient with two IgG subclasses was identical with the patient that exhibited
immunoreactivity to more than one domain. One single serum (patient PSY_203) yielded no
clearly interpretable results and was therefore excluded from the summary in Table 2. Overall,
no strict correlation of one of the subclasses with a single domain was observed. Demographical data of patients with synapsin autoantibodies The diagnoses, clinical and paraclinical findings of all synapsin-positive patients have been
published in our previous studies. In the current study 13 (eight females, five males) out of 20
patients exhibited immunoreactivity to fragments of synapsin Ia and had a mean ± SD (range)
age of 42,9 ± 19,8 (20–77) years. Controls (nine) had a mean ± SD (range) age of 38,6 ± 17,2
(18–65) years. Discussion Despite the apparent clinical heterogeneity, a preference in epitope specificity of anti synapsin
autoantibodies in psychiatric and neurological patients exists. Of the 13 patients showing a
clearly detectable immune reaction to one of the investigated synapsin fragments six sera
exhibited binding of IgG to the A-domain. The same number was detected for the D-domain. Binding to the B-domain and E-domain was shown for one individual patient. Concerning
IgG subclasses, IgG1, IgG3 and, with highest prevalence, IgG4 were detected. Intracellular synaptic proteins as autoimmune targets: Pathogenic
relevance Given the fact that synapsin is a ubiquitous presynaptic protein, occurrence of autoantibodies
in a variety of psychiatric and neurological disorders is conceivable. Subject to functional evi-
dence, it remains unclear if the existence of autoantibodies to synapsin in serum or CSF has a 8 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0208636
December 13, 2018 Autoantibodies and synapsin domains Fig 3. IgA antibodies to synapsin in serum and CSF of the previously described index patient with limbic
encephalitis are directed against the D-domain of synapsin. (A)HEK293 cells were transfected with eYFP-tagged
synapsin I fragment ABC, eGFP-tagged DE, D or E. Cells were incubated with patient serum or CSF at a dilution of
1:320. Bound patient IgA was detected using a Texas red-coupled secondary antibody directed against human IgA. Both the patient’s serum and CSF (Patient IgA) demonstrated positive immunoreactivity that co-localized with the
eGFP tag of synapsin DE and single D fragments, but with none of the other constructs. (B) Serum and CSF of two
healthy controls (control IgA) did not bind to the cells transfected in the same way. https://doi.org/10.1371/journal.pone.0208636.g003 bodies to synapsin in serum and CSF of the previously described index patient with limbic
( )
ll
f
d
h Fig 3. IgA antibodies to synapsin in serum and CSF of the previously described index patient with limbic
encephalitis are directed against the D-domain of synapsin. (A)HEK293 cells were transfected with eYFP-tagged
synapsin I fragment ABC, eGFP-tagged DE, D or E. Cells were incubated with patient serum or CSF at a dilution of
1:320. Bound patient IgA was detected using a Texas red-coupled secondary antibody directed against human IgA. Both the patient’s serum and CSF (Patient IgA) demonstrated positive immunoreactivity that co-localized with the
eGFP tag of synapsin DE and single D fragments, but with none of the other constructs. (B) Serum and CSF of two
healthy controls (control IgA) did not bind to the cells transfected in the same way. https://doi.org/10.1371/journal.pone.0208636.g003 https://doi.org/10.1371/journal.pone.0208636.g003 pathogenic relevance or represents an epiphenomenon. In addition, it remains to be elucidated
whether the antigenic epitope presentation of synapsin domains relies on the extracellular pre-
sentation of synapsin in the course of an ongoing neuronal repair mechanism as proposed in
an in vivo model [14], a specific extracellular presentation by a so far unknown surface trans-
port process or just results from neuronal cell loss. Intracellular synaptic proteins as autoimmune targets: Pathogenic
relevance (A) HEK293 cells were transfected with eGFP-tagged full length rat synapsin Ia and incubated with serum of
patient CIS117 at a dilution of 1:320. Bound antibodies of the IgG subclasses IgG1-4 were detected using subclass-specific Alexa
647-coupled secondary antibodies. The patient’s serum demonstrated positive immunoreactivity for IgG1 antibodies but none of the
other subclasses. (B) HEK293 cells were transfected with eGFP-tagged full length rat synapsin Ia and incubated with serum of
patient PSY 149 (upper two panels) and patient PSY 171 at a dilution of 1:320. Bound antibodies of the IgG subclasses IgG1-4 were
detected as given in (A). Serum of patient PSY 149 demonstrated positive immunoreactivity for IgG1 and IgG4 antibodies but none
of the other subclasses. Serum of patient PSY 171 demonstrated positive immunoreactivity for IgG3 antibodies but none of the other
subclasses. (C) Serum of a healthy control did not exhibit immunoreactivity for any of the IgG subclasses. https://doi org/10 1371/journal pone 0208636 g004 Fig 4. IgG subclasses of antibodies to synapsin in sera of patients with a clinically isolated syndrome (CIS), bipolar disorder
and depression. (A) HEK293 cells were transfected with eGFP-tagged full length rat synapsin Ia and incubated with serum of
patient CIS117 at a dilution of 1:320. Bound antibodies of the IgG subclasses IgG1-4 were detected using subclass-specific Alexa
647-coupled secondary antibodies. The patient’s serum demonstrated positive immunoreactivity for IgG1 antibodies but none of the
other subclasses. (B) HEK293 cells were transfected with eGFP-tagged full length rat synapsin Ia and incubated with serum of
patient PSY 149 (upper two panels) and patient PSY 171 at a dilution of 1:320. Bound antibodies of the IgG subclasses IgG1-4 were
detected as given in (A). Serum of patient PSY 149 demonstrated positive immunoreactivity for IgG1 and IgG4 antibodies but none
of the other subclasses. Serum of patient PSY 171 demonstrated positive immunoreactivity for IgG3 antibodies but none of the other
subclasses. (C) Serum of a healthy control did not exhibit immunoreactivity for any of the IgG subclasses. https://doi.org/10.1371/journal.pone.0208636.g004 https://doi.org/10.1371/journal.pone.0208636.g004 association was detectable in patients with autoantibodies to synapsin. Moreover, the index
patient clinically improved following immunotherapy indicating a functional relevance of anti
synapsin antibodies [1]. Hitherto, autoantibodies to intracellular synaptic proteins other than
synapsin have already been described for glutamate decarboxylase 65 (GAD65) and amphi-
physin in large patient cohorts [19; 20]. Intracellular synaptic proteins as autoimmune targets: Pathogenic
relevance Irrespective of how the immune system
gets access to synapsin, an intracellular action of antibodies might occur, as shown for autoan-
tibodies against neurofilament proteins [15]. In contrast to other known autoantibodies to
intracellular target proteins such as anti-Hu, anti-Ri or anti-Yo, [16; 17; 18] no strong tumor pathogenic relevance or represents an epiphenomenon. In addition, it remains to be elucidated
whether the antigenic epitope presentation of synapsin domains relies on the extracellular pre-
sentation of synapsin in the course of an ongoing neuronal repair mechanism as proposed in
an in vivo model [14], a specific extracellular presentation by a so far unknown surface trans-
port process or just results from neuronal cell loss. Irrespective of how the immune system
gets access to synapsin, an intracellular action of antibodies might occur, as shown for autoan-
tibodies against neurofilament proteins [15]. In contrast to other known autoantibodies to
intracellular target proteins such as anti-Hu, anti-Ri or anti-Yo, [16; 17; 18] no strong tumor PLOS ONE | https://doi.org/10.1371/journal.pone.0208636
December 13, 2018 9 / 14 Autoantibodies and synapsin domains Fig 4. IgG subclasses of antibodies to synapsin in sera of patients with a clinically isolated syndrome (CIS), bipolar disorder
and depression. (A) HEK293 cells were transfected with eGFP-tagged full length rat synapsin Ia and incubated with serum of
patient CIS117 at a dilution of 1:320. Bound antibodies of the IgG subclasses IgG1-4 were detected using subclass-specific Alexa
647-coupled secondary antibodies. The patient’s serum demonstrated positive immunoreactivity for IgG1 antibodies but none of the
other subclasses. (B) HEK293 cells were transfected with eGFP-tagged full length rat synapsin Ia and incubated with serum of
patient PSY 149 (upper two panels) and patient PSY 171 at a dilution of 1:320. Bound antibodies of the IgG subclasses IgG1-4 were
detected as given in (A). Serum of patient PSY 149 demonstrated positive immunoreactivity for IgG1 and IgG4 antibodies but none
of the other subclasses. Serum of patient PSY 171 demonstrated positive immunoreactivity for IgG3 antibodies but none of the other
subclasses. (C) Serum of a healthy control did not exhibit immunoreactivity for any of the IgG subclasses. https://doi org/10 1371/journal pone 0208636 g004 Fig 4. IgG subclasses of antibodies to synapsin in sera of patients with a clinically isolated syndrome (CIS), bipolar disorder
and depression. Autoantibodies and synapsin domains Table 2. Distinct IgG subclasses of antibodies to synapsin Ia. Patient
IgG1
IgG2
IgG3
IgG4
Target Domain
CIS117 (serum)
+
-
-
-
D
CIS117 (CSF)
+
-
-
-
D
PSY_0012
-
-
-
+
A
PSY_0032
-
-
-
+
A
PSY_0068
-
-
-
+
D
PSY_0094
-
-
-
+
D
PSY_0141
-
-
-
+
D
PSY_0149
+
-
-
+
A,B,D,E
PSY_0171
-
-
+
-
D
PSY_1851
-
-
-
+
A
PSY_1852
-
-
-
+
A
PSY_0189
-
-
+
-
A
Presence (+) or absence (-) of immunoreactivity is indicated. Differential attribution of IgG1 and IgG4 antibodies to individual domains is unknown. Table 2. Distinct IgG subclasses of antibodies to synapsin Ia. Presence (+) or absence (-) of immunoreactivity is indicated. Differential attribution of IgG1 and IgG4 antibodies to individual domains is unknown. https://doi.org/10.1371/journal.pone.0208636.t002 encephalitis) than antibodies to the A-domain, that dominated in bipolar disorder patients
(four out of six patients). The relatively small number of patients, however, prevents us from
drawing definite conclusions on the association of specific diseases with antibodies to distinct
synapsin domains. In addition to our previous prevalence study that addressed the presence of antibodies in
serum we had one additional CSF sample available for testing (CIS_0117). As shown for IgA
antibodies of the index patient with limbic encephalitis this CSF also showed IgG reactivity to
the D-domain. Future studies are needed to show whether the D-domain is the preferred tar-
get of CSF antibodies. Noteworthy, for both available CSF samples with antibodies to synapsin
an intrathecal synthesis of immunoglobulins could be detected in this study and [1]. Antibodies to full length synapsin not reacting with single domains or
domain stretches The observation that seven out of 20 sera reacting with full-length synapsin Ia did not react
with distinct synapsin domains might be explained by the fact that a certain conformational
epitope structure can be provided only by the full-length protein. Missing homology between
human and rat sequences can be ruled out to account for this observation since all 20 sera
reacted with full length rat synapsin. Alternatively, anti synapsin antibodies may exhibit a
lower affinity to the expressed domains than to the full-length protein and therefore escaped
detection in our assay, that was standardized to yield a high signal to noise ratio based on a
1:320 dilution of the applied patient samples. Antibody titers to full length protein were ini-
tially analysed for all individual sera [2] but can not be used to predict the binding probability
to individual domains since sera with a titer range of 1:320–1:100.000 bound to synapsin frag-
ments. Folding differences between the individual synapsin constructs might also account for
the observation that two sera exclusively reacted with either the DE fragment or the D-domain
but not with both constructs as expected. Intracellular synaptic proteins as autoimmune targets: Pathogenic
relevance For these two autoantibodies associated with different
neurological diseases like stiff-man syndrome evidence for a pathogenic potential was sug-
gested by rodent animal models showing induction of anxiety by transfer of stiff-person IgG
[21] or a reduced GABAergic inhibition by amphiphysin IgG [22]. Internalisation of amphi-
physin antibodies was shown recently to reduce the presynaptic vesicle pool, adding to mecha-
nistic understanding of a suggested pathogenic effect [23]. In the case of synapsin, a putative
pathogenic mechanism of antibody binding might rely on the interference with the phosphor-
ylation status of synapsin, a modification crucially involved in regulating its function and
shown to affect residues both of the A and the D domain regions [24]. association was detectable in patients with autoantibodies to synapsin. Moreover, the index
patient clinically improved following immunotherapy indicating a functional relevance of anti
synapsin antibodies [1]. Hitherto, autoantibodies to intracellular synaptic proteins other than
synapsin have already been described for glutamate decarboxylase 65 (GAD65) and amphi-
physin in large patient cohorts [19; 20]. For these two autoantibodies associated with different
neurological diseases like stiff-man syndrome evidence for a pathogenic potential was sug-
gested by rodent animal models showing induction of anxiety by transfer of stiff-person IgG
[21] or a reduced GABAergic inhibition by amphiphysin IgG [22]. Internalisation of amphi-
physin antibodies was shown recently to reduce the presynaptic vesicle pool, adding to mecha-
nistic understanding of a suggested pathogenic effect [23]. In the case of synapsin, a putative
pathogenic mechanism of antibody binding might rely on the interference with the phosphor-
ylation status of synapsin, a modification crucially involved in regulating its function and
shown to affect residues both of the A and the D domain regions [24]. When comparing the clinical diagnoses, antibodies to the D-domain were observed in a
somewhat broader spectrum of diseases (depression, bipolar disorder, ADHD, CIS, limbic PLOS ONE | https://doi.org/10.1371/journal.pone.0208636
December 13, 2018 10 / 14 Acknowledgments The authors thank the participants of this study for donating blood and Birgit Metze and Mar-
ion Mo¨bes for excellent technical assistance. The authors also thank Biobank1 for handling,
registering, and storing all blood samples from the Norwegian cohort. Autoantibodies and synapsin domains being the last prevalent subclass detected in serum [25] IgG4 antibodies therefore seem to be
the dominant, albeit not exclusive, subclass in synapsin autoimmunity, at least for the relatively
few cases described so far. So, the ability to fix complements seems not to be necessary for the
immune reaction in the majority of patients. Antibodies of the IgG4 subclass have already been
described for LGI1 and Caspr2 encephalitis [26; 27], and DPPX encephalitis [28]. Moreover,
this subclass is associated with an IgG4-related systemic disease, able to cause pancreatitis and
infection of other organs [29, 30]. In summary, the present study adds data to further characterize the recently identified anti-
neuronal antibodies against synapsin in molecular detail. Functional cell culture and animal
studies investigating the pathological potential of these antibodies by electrophysiological and
biochemical methods are under way to shed light on the putative pathogenic impact of these
antibodies. Writing – review & editing: Gudrun Ahnert-Hilger, Klemens Ruprecht. Writing – review & editing: Gudrun Ahnert-Hilger, Klemens Ruprecht. Author Contributions Conceptualization: Fabio Benfenati, Gudrun Ahnert-Hilger, Klemens Ruprecht, Markus
Ho¨ltje. Conceptualization: Fabio Benfenati, Gudrun Ahnert-Hilger, Klemens Ruprecht, Markus
Ho¨ltje. Investigation: Robert Mertens, Sarah Melchert, Daniel Gitler, Johannes Piepgras, Elena
Kochova. Resources: Morten Brix Schou, Sverre Georg Saether, Arne Vaaler, Fabio Benfenati, Klemens
Ruprecht. Resources: Morten Brix Schou, Sverre Georg Saether, Arne Vaaler, Fabio Benfenati, Klemens
Ruprecht. Resources: Morten Brix Schou, Sverre Georg Saether, Arne Vaaler, Fabio Benfenati, Klemens
Ruprecht. Supervision: Markus Ho¨ltje. Supervision: Markus Ho¨ltje. Writing – original draft: Markus Ho¨ltje. Distinct IgG subclasses Of the four IgG subclasses described in the literature we were able to detect IgG1 (two
patients), IgG3 (two patients) and, most frequently, IgG4 (eight patients) in our cohort. Despite 11 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0208636
December 13, 2018 References 1. Piepgras J., Ho¨ltje M., Otto C., Harms H., Satapathy A., Cesca F., et al. Intrathecal immunoglobulin A
and G antibodies to synapsin in a patient with limbic encephalitis. Neurol Neuroimmunol Neuroinflamm. 2015 Nov 4; 2(6):e169. https://doi.org/10.1212/NXI.0000000000000169 PMID: 26587554 2. Ho¨ltje M., Mertens R., Kochova E., Brix Schou M., Saether S.G., Pru¨ss H., et al. Synapsin-antibodies in
psychiatric and neurological disorders: prevalence and clinical finding. Brain, Behaviour and Immunity
2017; Jul 18. pii: S0889-1591(17)30218-0. https://doi.org/10.1016/j.immuni.2017.05.003 3. Fassio A., Patry L., Congia S., Onofri F., Piton A., Gauthier J., et al. Syn1 loss-of-function mutations in
autism and partial epilepsy cause impaired synaptic function. Hum Mol Genet 2011; 20:2297–307. https://doi.org/10.1093/hmg/ddr122 PMID: 21441247 4. Baldelli P., Fassio A., Valtorta F., Benfenati F. Lack of synapsin I reduces the readily releasable pool of
synaptic vesicles at central inhibitory synapses. Journal of Neuroscience 2007; 27:13520–13531. https://doi.org/10.1523/JNEUROSCI.3151-07.2007 PMID: 18057210 5. Bragina L., Candiracci C., Barbaresi P., Giovedı` S., Benfenati F., Conti F. Heterogeneity of glutamater-
gic and GABAergic release machinery in cerebral cortex. Neuroscience 2007; 146:1829–1840. https://
doi.org/10.1016/j.neuroscience.2007.02.060 PMID: 17445987 6. Gitler D., Xu Y., Kao H.T., Lin D., Lim S., Feng J., et al. Molecular determinants of synapsin targeting to
presynaptic terminals. J Neurosci 2004; 24, 3711–3720. https://doi.org/10.1523/JNEUROSCI.5225-
03.2004 PMID: 15071120 12 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0208636
December 13, 2018 Autoantibodies and synapsin domains 7. Hilfiker S., Benfenati F., Doussau F., Nairn A.C., Czernik A.J., Augustine G.J., et al. Structural domains
involved in the regulation of transmitter release by synapsins. Journal of Neuroscience 2005; 25:2658–
2669. https://doi.org/10.1523/JNEUROSCI.4278-04.2005 PMID: 15758176 8. Cheetham J.J., Hilfiker S., Benfenati F., Weber T., Greengard P., Czernik A.J. Identification of synapsin
I peptides that insert into lipid membranes. Biochem J 2001; 354:57–66. PMID: 11171079 9. Gerth F., Ja¨pel M., Pechstein A., Kochlamazashvili G., Lehmann M., Puchkov D., et al. Intersectin asso-
ciates with synapsin and regulates its nanoscale localization and function. Proc Natl Acad Sci U S A. 2017 Nov7; 114(45):12057–12062. https://doi.org/10.1073/pnas.1715341114 PMID: 29078407 10. Schou M., Sæther S.G., Borowski K., Teegen B., Kondziella D., Stoecker W., et al. Prevalence of
serum anti-neuronal autoantibodies in patients admitted to acute psychiatric care. Psychol Med. 2016
Dec; 46(16):3303–3313. https://doi.org/10.1017/S0033291716002038 PMID: 27609625 11. Vaccaro P., Dente L., Onofri F., Zucconi A., Martinelli S., Valtorta F, et al. Anti-synapsin monoclonal
antibodies: epitope mapping and inhibitory effects on phosphorylation and Grb2 binding. Brain Res Mol
Brain Res. 1997 Dec 1; 52(1):1–16. PMID: 9450672 12. References Reiber H., Lange P. Quantification of virus-specific antibodies in cerebrospinal fluid and serum: sensi-
tive and specific detection of antibody synthesis in brain. Clin Chem 1991; 37:1153–1160. PMID:
1855284 13. Su¨dhof T C. The structure of the human synapsin I gene and protein. J Biol Chem. 1990 May 15; 265
(14):7849–52. PMID: 2110562 14. Wang S., Cesca F., Loers G., Schweizer M., Buck F., Benfenati F, et al. Synapsin I is an oligomannose-
carrying glycoprotein, acts as an oligomannose-binding lectin, and promotes neurite outgrowth and neu-
ronal survival when released via glia-derived exosomes. J Neurosci. 2011 May 18; 31(20):7275–90. https://doi.org/10.1523/JNEUROSCI.6476-10.2011 PMID: 21593312 15. Congdon E.E., Gu J., Sait H.B., Sigurdsson E.M. Antibody uptake into neurons occurs primarily via cla-
thrin-dependent Fcgamma receptor endocytosis and is a prerequisite for acute tau protein clearance. J
Biol Chem 2013; 288, 35452–35465. https://doi.org/10.1074/jbc.M113.491001 PMID: 24163366 16. Graus F., Keime-Guibert F., Reñe R., Benyahia B., Ribalta T., Ascaso C., et al. Anti-Hu-associated
paraneoplastic encephalomyelitis: analysis of 200 patients. Brain. 2001 Jun; 124(Pt 6):1138–48. PMID:
11353730 17. Pittock S.J., Lucchinetti C.F., Lennon V.A. Anti-neuronal nuclear autoantibody type 2: paraneoplastic
accompaniments.Ann Neurol. 2003 May; 53(5):580–7. https://doi.org/10.1002/ana.10518 PMID:
12730991 18. Peterson K., Rosenblum M.K., Kotanides H., Posner J.B. Paraneoplastic cerebellar degeneration. I. A
clinical analysis of 55 anti-Yo antibody-positive patients.Neurology. 1992 Oct; 42(10):1931–7. PMID:
1407575 19. Pittock S.J., Lucchinetti C.F., Parisi J.E., Benarroch E.E., Mokri B., Stephan C.L., et al. Amphiphysin
autoimmunity: paraneoplastic accompaniments. Ann Neurol. 2005 Jul; 58(1):96–107. https://doi.org/
10.1002/ana.20529 PMID: 15984030 20. Pittock S.J., Yoshikawa H., Ahlskog J.E., Tisch S.H,. Benarroch E.E., Kryzer T.J., et al. Glutamic acid
decarboxylase autoimmunity with brainstem, extrapyramidal, and spinal cord dysfunction. Mayo Clin
Proc. 2006 Sep; 81(9):1207–14. https://doi.org/10.4065/81.9.1207 PMID: 16970217 21. Geis C., Weishaupt A., Gru¨newald B., Wultsch T., Reif A., Gerlach M., et al. Human stiff-person syn-
drome IgG induces anxious behavior in rats. PLoS One. 2011 Feb 8; 6(2):e16775. https://doi.org/10. 1371/journal.pone.0016775 PMID: 21346811 22. Geis C., Weishaupt A., Hallermann S., Gru¨newald B., Wessig C., Wultsch T., et al. Stiff person syn-
drome-associated autoantibodies to amphiphysin mediate reduced GABAergic nhibition. Brain. 2010
Nov; 133(11):3166–80. https://doi.org/10.1093/brain/awq253 PMID: 20884644 23. Werner C., Pauli M., Doose S., Weishaupt A., Haselmann H., Gru¨newald B., et al. Humanautoantibo-
dies to amphiphysin induce defective presynaptic vesicle dynamics and composition. Brain. 2016 Feb;
139(Pt 2):365–7 https://doi.org/10.1093/brain/awv324 PMID: 26582558 24. Su¨dhof T.C., Czernik A.J., Kao H.T., Takei K., Johnston P.A., Horiuchi A., et al. PLOS ONE | https://doi.org/10.1371/journal.pone.0208636
December 13, 2018 27.
van Sonderen A., Ariño H., Petit-Pedrol M., Leypoldt F., Ko¨rtve´lyessy P., Wandinger K.P., et al. The
clinical spectrum of Caspr2 antibody-associated disease. Neurology. 2016 Aug 2; 87(5):521–8. https://
doi.org/10.1212/WNL.0000000000002917 PMID: 27371488 References Synapsins: mosaics of
shared and individual domains in a family of synaptic vesicle phosphoproteins. Science. 1989 Sep 29;
245(4925):1474–80. PMID: 2506642 25. Engelhart S., Glynn R.J., Schur P.H. Disease associations with isolated elevations of each of the four
IgG subclasses. Semin Arthritis Rheum. 2017 Oct; 47(2):276–280. https://doi.org/10.1016/j.semarthrit. 2017.03.021 PMID: 28457528 26. Ariño H., Armangue´ T., Petit-Pedrol M., Sabater L., Martinez-Hernandez E., Hara M., et al. Anti-LGI1-
associated cognitiveimpairment: Presentation and long-term outcome. Neurology. 2016 Aug 23; 87
(8):759–65. https://doi.org/10.1212/WNL.0000000000003009 PMID: 27466467 13 / 14 PLOS ONE | https://doi.org/10.1371/journal.pone.0208636
December 13, 2018 Autoantibodies and synapsin domains 27. van Sonderen A., Ariño H., Petit-Pedrol M., Leypoldt F., Ko¨rtve´lyessy P., Wandinger K.P., et al. The
clinical spectrum of Caspr2 antibody-associated disease. Neurology. 2016 Aug 2; 87(5):521–8. https://
doi.org/10.1212/WNL.0000000000002917 PMID: 27371488 28. Hara M., Ariño H., Petit-Pedrol M., Sabater L., Titulaer M.J., Martinez-Hernandez E., et al. DPPX anti-
body-associated encephalitis: Main syndrome and antibody effects. Neurology. 2017 Apr 4; 88
(14):1340–1348. https://doi.org/10.1212/WNL.0000000000003796 PMID: 28258082 29. Hamano H., Kawa S., Horiuchi A., Unno H., Furuya N., Akamatsu T., et al. High serum IgG4 concentra-
tions in patients with sclerosing pancreatitis. N Engl J Med. 2001 Mar 8; 344(10):732–8. https://doi.org/
10.1056/NEJM200103083441005 PMID: 11236777 30. Hamano H., Tanaka E., Ishizaka N., Kawa S. IgG4-related Disease—A Systemic Disease that
Deserves Attention Regardless of One’s Subspecialty. Intern Med. 2018 May 1; 57(9):1201–1207. https://doi.org/10.2169/internalmedicine.9533-17 PMID: 29279491 14 / 14
|
https://openalex.org/W1968176131
|
https://www.scielo.br/j/aob/a/DhvRLv7mwrBDgLXcjLftNfq/?lang=pt&format=pdf
|
English
| null |
Estudo macroscópico e histológico de reparos osteocondrais biologicamente aceitáveis
|
Acta Ortopédica Brasileira
| 2,004
|
cc-by
| 6,174
|
Estudo macr
Estudo macr
Estudo macr
Estudo macr
Estudo macroscópico e histológico de r
oscópico e histológico de r
oscópico e histológico de r
oscópico e histológico de r
oscópico e histológico de repar
epar
epar
epar
eparos
os
os
os
os
osteocondrais biologicamente aceitáveis.
osteocondrais biologicamente aceitáveis.
osteocondrais biologicamente aceitáveis.
osteocondrais biologicamente aceitáveis.
osteocondrais biologicamente aceitáveis. JOSÉ LEÃO RIBEIRO1, GILBERTO LUIS CAMANHO2, LUIZ CARLOS TAKITA3 ARTIGO ORIGINAL / ORIGINAL ARTICLE ARTIGO ORIGINAL / ORIGINAL ARTICLE INTRODUCTION The surgical treatment of the chondral and osteochondral injuries
that affect articulations that support load, mainly the knee, still repre-
sents a challenge for the orthopedist. This is due to the characteristics
of the articular hyaline cartilage, that is unprovided of vascularization
and so has limited potential of cicatrization. According to Mankim(15),
the cicatrization of an injury restricted to the articular hyaline cartilage
(chondral defect) doesn’t follow the three natural phases - the necro-
sis, the inflammation and the repair - exactly because of its avascular
condition. As in any other injured body tissue, the same phase of initial
necrosis occurs and consequently there is loss of cells and matrix. As
the chondrocytes are relatively insensitive to the condition of installed
hypoxia; there is a smaller number of dead cells than in any other or-
ganical tissue. A second phase of inflammation, as is entirely mediat-
ed for the vascular system, is absent. Consequently, the hematoma
formation doesn’t occur, and so it does not produce fibrin or coagulum
of fibrin that would work as a frame for the new repair tissue growth. SUMMARY Este estudo teve como finalidade avaliar macroscopicamente e
histológicamente defeitos osteocondrais já cicatrizados, também co-
nhecidos como reparos. Foram utilizados seis coelhos machos, adul-
tos, albinos da raça Nova Zelândia. Defeitos cilíndricos osteocon-
drais de 3.2 mm de diâmetro por 4.0 mm de profundidade foram
criados cirurgicamente em ambos côndilos femorais mediais. O ci-
lindro osteocondral retirado do joelho esquerdo (defeito não tratado)
foi implantado no joelho direito (joelho tratado). Comparou-se ma-
croscopicamente e histológicamente ambos tipos de defeitos após
doze semanas de evolução. A avaliação macroscópica de todos os
defeitos mostrou evolução para reparos denominados biologicamente
aceitáveis. O termo “biologicamente aceitável” foi utilizado para defi-
nir reparos, que à observação macroscópica, se apresentaram como
tecido neo-formado semelhante à fibrocartilagem, brilhante, liso, fir-
me, em continuidade com a cartilagem adjacente. Como todos os
defeitos, tratados e não tratados, eram macroscopicamente seme-
lhantes, realizou-se um estudo histológico comparativo para averi-
guar qual tipo de tecido de reparação se formava em ambos defei-
tos. Pela análise histológica dos reparos biologicamente aceitáveis,
concluiu-se que houve formação de tecido cartilaginoso hialino nos
defeitos tratados com enxerto autólogo e de tecido fibrocartilaginoso
nos defeitos não tratados. The aim of this study was to evaluate macroscopically and histo-
logically healed osteochondral defects, also known as repairs. Six adult,
male, New Zealander White rabbits were used. Cylindrical osteochon-
dral defects of 3.2 mm in diameter by 4.0 mm in depth were artificially
created in the load-bearing surfaces of both medial femoral condyles. The osteochondral graft collected from the left knee (untreated defect)
was implanted in the right knee (treated defect). Both defects were
compared macroscopically ad histologically after twelve weeks. The
macroscopic evaluation of all defects demostrated evolution in repairs
called “biologically acceptable”. The “biologically acceptable” term was
used to define repairs, that at macrocospic observation, presented as
neo-formed tissue similar to fibrocartilage, brilliant, smooth, firm, in
continuity with the adjacent cartilage. As all the defects, treated and
untreated, were macroscopically similar, a histological comparative
study was made to verify which type of repair tissue was formed in the
surface of both defects. By histologic analysis of the biologically ac-
ceptable repairs, the authors conclude that: treated defects with autol-
ogous grafts were filled with hyaline cartilaginous tissue and untreated
defects were filled with fibrocartilaginous tissue. Key words: Knee; Autologous transplantation; Articular cartilage; Os-
teochondral defect; Rabbit Descritores: Joelho; Transplante autólogo; Cartilagem articular; De-
feito osteocondral; Coelho Trabalho recebido em 21/05/03. Aprovado em 16/02/04.
Trabalho realizado no Serviço de Ortopedia do Departamento de Clinica
Cirúrgica da Universidade Federal de Mato Grosso.
1 - Professor Adjunto de Ortopedia da Universidade Federal de Mato
Grosso
2 - Professor Associado da Faculdade de Medicina da Universidade de
São Paulo - FMUSP
3 - Professor Assistente de Anatomia Patológica Especial da Universidade
Federal de Mato Grosso do Sul
Endereço para correspondência: R. Osório Duque Estrada, 15 – Hospital
Ortopédico, CEP -78005-720, Cuiabá-MT, e-mail: drleao@terra.com.br Trabalho realizado no Serviço de Ortopedia do Departamento de Clinica
Cirúrgica da Universidade Federal de Mato Grosso. Work performed at the Orthopedics Service of the Surgical Clinical
Department of the Universidade Federal de Mato Grosso
1 – Adjuct Professor of Orthopedics of the Universidade Federal de Mato
Grosso
2 – Associate Professor of the Universidade de São Paulo - FMUSP
3 – Assistant Professor of Special Pathological Anatomy of the
Universidade Federal de Mato Grosso do Sul
Mail address: R. Osório Duque Estrada, 15 – Hospital Ortopédico, CEP -
78005-720, Cuiabá-MT, e-mail: drleao@terra.com.br Work performed at the Orthopedics Service of the Surgical Clinical
Department of the Universidade Federal de Mato Grosso INTRODUÇÃO O tratamento cirúrgico das lesões condrais e osteocondrais que
acometem articulações que suportam carga, principalmente o joe-
lho, ainda representa um desafio para o ortopedista. Isto se deve às
características da cartilagem hialina articular, que por ser desprovida
de vascularização tem limitado potencial de cicatrização. De acordo com Mankim(15), a cicatrização de uma lesão restrita à
cartilagem hialina articular (defeito condral), não obedece inteira-
mente às três fases naturais que são a necrose, inflamação e repara-
ção, justamente por causa da sua condição avascular. Como em qualquer outro tecido corpóreo lesado, existe a mes-
ma fase de necrose inicial e conseqüentemente ocorre a perda celu-
lar e da matriz. Como os condrócitos são relativamente insensíveis à
condição de hipóxia instalada, existe um número menor de células 2 – Associate Professor of the Universidade de São Paulo - FMUSP
3 – Assistant Professor of Special Pathological Anatomy of the
Universidade Federal de Mato Grosso do Sul Mail address: R. Osório Duque Estrada, 15 – Hospital Ortopédico, CEP -
78005-720, Cuiabá-MT, e-mail: drleao@terra.com.br 16 ACTA ORTOP BRAS 12(1) - JAN/MAR, 2004 This way there isn’t continuity in this process. Considering the third
phase that is the repair, the absence of a vascular or inflammatory phase
limits the number of available cells to respond to the trauma and the
repair capacity fails in the remaining chondrocytes, blocking the cica-
trization process definitively. mortas do que em qualquer outro tecido orgânico. A segunda fase
de inflamação por estar inteiramente mediada pelo sistema vascular
é ausente, conseqüentemente não há formação de hematoma, não
há produção de fibrina e nem de coágulo de fibrina que serviria como
arcabouço para o novo tecido de reparação que iria crescer, não
havendo conseqüentemente continuidade do processo. Consideran-
do a terceira fase que é a reparação, a ausência de uma fase vascu-
lar ou inflamatória limita consideravelmente o número de células dis-
poníveis para responder ao trauma e a capacidade de reparo falha
nos condrócitos remanescente o que termina por bloquear definitiva-
mente o processo de cicatrização. However it is known that if the injury extends until the subchondral
bone (osteochondral defect), that is well vascularized, all the three
phases occur naturally(3,5,11,15). The hematoma is quickly organized with
coagulum of fibrin, white globules and elements of the bone marrow. INTRODUÇÃO Undifferentiated cells of the marrow and vascular endothelium are trans-
formed into primitive fibroblasts and become fibroblastic vascularized
tissues of repair by receiving capillaries and coagulum of fibrin(15). The
study of the cicatrization of the cartilaginous injuries has been per-
formed in diverse animal experimental models. Probably, no other ex-
periment has been so frequently performed, concerning the study of
cartilage, experimental surgical injury and the observation of the carti-
lage behavior by macroscopical, radiological, biochemical, biomechan-
ic, biomolecular and ultrastructural view(3,4,7,15). The focal injuries of the
articular cartilage require surgical treatment that includes the classic
methods of stimulation of the bone marrow such as debridement,
multiple perforations, abrasions, microfractures(12,19,22,23), and modern
biological methods such as periosteal and perichondral transplants,
implantation of cultivated autologous chondrocytes, and autologous
osteochondral grafts(1,9,17,18,21). Because of the high cost and technical
restrictions imposed to some biological methods of treatment of the
osteochondral injuries, some authors have also searched efficient
methods and easier to perform. No entanto é reconhecido que se a lesão se estende até o osso
subcondral (defeito osteocondral), muito bem vascularizado, todas
as três fases ocorrem naturalmente(3,5,11,15). O hematoma rapidamen-
te se organiza com coágulos de fibrina, glóbulos brancos e elemen-
tos da medula óssea. Células indiferenciadas da medula e endotélio
vascular são transformadas em fibroblastos primitivos que com o
aporte de capilares e coágulos de fibrina se transformam em tecido
fibroblástico vascularizado de reparação(15). O estudo da cicatrização das lesões cartilaginosas tem sido efe-
tuado em diversos modelos experimentais animais. Provavelmente
nenhum outro experimento tenha sido executado tão freqüentemen-
te, quando se trata de estudo sobre cartilagem, quanto a lesão cirúr-
gica experimental e a observação do comportamento da cartilagem
do ponto vista macroscópico, radiológico, bioquímico, biomecânico,
biomolecular e ultraestrutural(3,4,7,15). As lesões focais da cartilagem articular requerem tratamento ci-
rúrgico que vão desde métodos clássicos de estimulação da medula
óssea como desbridamento, perfurações múltiplas, abrasões, mi-
crofraturas(12,19,22,23), até métodos biológicos modernos como trans-
plantes periosteais e pericondriais, implante de condrócitos autólo-
gos cultivados, e enxertos autólogos osteocondrais(1,9,17,18,21). Por causa
do alto custo e de restrições técnicas impostas a vários métodos
biológicos de tratamento das lesões osteocondrais, alguns autores
buscaram métodos também eficazes, porem de maior facilidade de
execução. Um desses métodos simples e direto é sem dúvida o trans-
plante ou enxerto autólogo osteocondral que representa uma boa
alternativa para reparação biológica da lesão focal da superfície arti-
cular. MATERIAL AND METHOD Este modelo experimental teve como base o método de trata-
mento das lesões osteocondrais descrito e desenvolvido por Hango-
dy(9). Optou-se por provocar cirurgicamente defeitos osteocondrais
em ambos côndilos femorais de coelhos. O defeito do joelho direito
(D) foi preenchido por enxerto osteocondral autólogo (tratado) e o
defeito do joelho esquerdo (E) permaneceu vazio (não tratado). INTRODUÇÃO Consiste em preencher o defeito com cilindro osteocondral
único, ou conforme o tamanho da lesão, com uma série de peque-
nos enxertos osteocondrais, técnica que ficou conhecida como mo-
saicoplastia. Foi utilizada em cães, cavalos e cadáveres humanos
por Hangody(9) desde 1991 e as áreas doadoras e receptoras foram
estudadas para verificação do comportamento das lesões e sobre-
vida da cartilagem hialina transplantada. One of these simple and direct methods is the transplant or autol-
ogous osteochondral graft that represents a good alternative for bio-
logical repair of the focal injury of the articular surface. It consists of
filling the defect with a unique osteochondral cylinder, or according to
the size of the injury, with a series of small osteochondral grafts. This
technique is known as mosaicplasty. It has been used in dogs, horses
and human corpses by Hangody(9) since 1991 and the donor and re-
ceiver portions have been studied in order to verify the injuries behav-
ior and supervene of the transplanted hyaline cartilage. The method of treatment of the osteochondral injuries described
and developed by Hangody(9) was the base for this experimental mod-
el. Osteochondral defects in both femoral condyles of rabbits were
surgically produced. The defect of the right knee (D) was filled by au-
tologous osteochondral graft (treated) and the defect of the left knee
(E) remained empty (untreated). The aim of this study was to compare
macroscopically and histologically the osteochondral defects healed
after twelve weeks of evolution. 2 - Surgical technique As a pre-anesthetical drug, each animal received 0,2 mg/kg of
weight of acepromazine 1% by intramuscular. The anesthesia consist-
ed of the intramuscular application of hydrochloride of ketamine 5% in
the dose of 50 mg/kg of weight associated with xylazine 2% with the
dose of 3mg/kg of weight. 1 - Animals of experimentation Six male, albinic, adult rabbits of the New Zealander race, weight
between 3.4 kg and 3,6 kg were used in this study. The animals were
lodged in appropriate room and confined in metallic cages measuring
70X70X70. They were fed with standard pelleted ration and water ad
libitum. O objetivo deste estudo foi comparar macroscopicamente e his-
tológicamente os defeitos osteocondrais cicatrizados após doze se-
manas de evolução. ACTA ORTOP BRAS 12(1) - JAN/MAR, 2004 4.1 - Macroscopic evaluation Foram retirados ambos joelhos, examinados cuidadosamente
para anotação dos dados da avaliação macroscópica dos reparos
obtidos. As peças foram então encaminhadas para estudo histológi-
co. Five parameters were analyzed through the macroscopic inspec-
tion: the repair concerns the defect cicatrization, the continuity is the
levelling and integration of the repair with the adjacent cartilage. The other three parameters show the external characteristics of
the repair and they are the surface, the brightness and the consistency
of the tissue formed. A considered good result (biologically accept-
able) should present the five satisfactory parameters. On the other hand,
a considered not good result (biologically not acceptable) would
present one or more of these parameters not satisfactory. 1 - Animais de experimentação Para o estudo proposto foram utilizados seis coelhos machos,
albinos,adultos da raça Nova Zelândia pesando entre 3.4 kg e 3.6 kg. Os animais foram alojados em sala apropriada e confinados em gai-
olas metálicas medindo 70X70X70, alimentados com ração padrão
peletizada e água ad libitum. After the trichotomy of the surgical area it was performed a parap-
atellar medial incision followed by arthrotomy and lateral dislocation of
patella. 17 ACTA ORTOP BRAS 12(1) - JAN/MAR, 2004 Figura 1 - Enxerto osteocondral retirado do joelho
esquerdo – defeito não tratado. Figure 1 - Osteochondral graft removed from left knee –
untreated defect. 3 - Eutanásia Specifically for the proposed study, the analysis of two criteria of
evaluation was considered: macroscopic and histologic. Foi realizada nos animais aos 90 dias de evolução com aneste-
sia similar à do pré-operatório, seguida de injeção endovenosa de 60
mg/kg de cloreto de potássio. 4.1 - Avaliação macroscópica Analisou-se através da inspeção macroscópica cinco parâme-
tros: a reparação diz respeito a cicatrização propriamente dita do
defeito, a continuidade é a nivelação e integração do reparo com a
cartilagem adjacente. Os outros três parâmetros mostram as carac-
terísticas externas do reparo e são: a superfície, o brilho e a consis-
tência do tecido formado. Um resultado considerado bom (biologi-
camente aceitável) teria os cinco parâmetros satisfatórios e um re-
sultado não bom (biologicamente não aceitável) teria um ou mais
desses parâmetros não satisfatórios. 3 - Euthanasia It was performed in the animals af-
ter 90 days of evolution by a similar an-
esthesia used in the preoperative, fol-
lowed by endovenous injection of 60 mg/
kg of potassium chloride. O enxerto osteocondral retirado do
joelho E (defeito não tratado) foi então
transferido para o joelho D (defeito tra-
tado) (Figura 2). Procedeu-se a sínte-
se das feridas operatórias e os animais
foram devolvidos às suas respectivas
gaiolas para recuperação pós-operató-
ria. Both knees were removed in order
to be carefully examined. All the data of
the macroscopic evaluation of the ob-
tained repairs were noted. The parts then
were led to histologic study. Figura 2 - Enxerto osteocondral retirado do joelho direito –
defeito tratado. Figure 2 - Osteochondral graft removed from right knee –
treated defect. 4.2 - Avaliação Histológica Após a eutanásia a porção distal do fêmur foi removida em blo-
co, fixada em formol a 10%, descalcificada em solução de acido nitri-
co e formalina e estudada histologicamente. As lâminas foram cora-
das com Hematoxilina-Eosina. em secções no plano sagital, conten-
do o longo eixo do fêmur distal para melhor visibilização da extensão
do defeito. 2 - Técnica cirúrgica As the knee was flexed, the femoral
medial condyle was exposed and with
the help of a metallic trephine an osteo-
chondral cylinder of 3.2 mm/4.0 mm was
removed from the knee E (Figure 1). Through sequential perforations with up
to 3.2-mm drills, the osteochondral de-
fect in knee D was created. Como droga pré-anestésica cada
animal recebeu 0.2 mg/kg de peso de
acepromazina 1% via intramuscular. A
anestesia constou da aplicação por via
intramuscular de cloridrato de ketami-
na 5% na dose de 50 mg/kg de peso
associada a xilazina 2% na dose de
3mg / kg de peso. Osteochondral grafts removed from
knee E (untreated defect) were then
transferred to knee D (treated defect)
(Figure 2). The synthesis of the surgical
wounds was performed and the animals
returned to their respective cages for
postoperative recovery. Após tricotomia da área cirúrgica
realizou-se incisão parapatelar medial
seguida de artrotomia e luxação lateral
da patela. Com o joelho fletido expôs-
se o côndilo femoral medial e com au-
xílio de trefina metálica retirou-se um ci-
lindro osteocondral de 3.2 mm/4.0 mm
do joelho E (Figura 1). Através de per-
furações seqüenciais com brocas até
3.2 mm criou-se o defeito osteocondral
no joelho D. Figura 1 - Enxerto osteocondral retirado do joelho
esquerdo – defeito não tratado. Figure 1 - Osteochondral graft removed from left knee –
untreated defect. Figura 2 - Enxerto osteocondral retirado do joelho direito –
defeito tratado. Figure 2 - Osteochondral graft removed from right knee –
treated defect. 4.2 - Histologic evaluation After the euthanasia the distal portion of femur was removed in
block, settled in formol 10%, decalcificated in solution of nitric acid
and formalin and studied histologically. The blades were tinged with
Hematoxylin-Eosine. in cuts in the sagittal plan, including the long axle
of distal femur for a better view of the extension of the defect. In the histologic examination of the cuts of distal femur, the atten-
tion was particularly directed to the following elements: 1- the type of
regenerated tissue in the surface of the defect: cartilage, fibrocartilage
or fibrous tissue; 2- the appearance of the cartilaginous surface: smooth,
depressed or irregular; 3- presence or absence of the regenerated
subchondral bone. 4 - Critérios de avaliação Considerou-se especificamente para o estudo proposto a análi-
se de dois critérios de avaliação: macroscópico e histológico. 2 - Histologic aspect 0
00
00
100
19
95
0
01
05
0
00
00
0
00
00
eqüência de resultados na
cicatrização dos defeitos. frequency of results in the
n of defects healing. The histologic repair would only be con-
sidered satisfactory if the surface of the tis-
sue regenerated smoothly, were covered by
cartilage were in the same level of the adja-
cent cartilage. The differences in the types
of cicatrization tissue of the repairs were
clear. For the treated defects the surface of
the repairs presented the preservation of the
smooth, regular original hyaline cartilage and
in the same level of the adjacent hostess
cartilage. The deep part of the repair be-
came bone tissue with trabecular standard
close to normality (Figure 3-B). The surface
of the repairs proceeding from treated de-
fects was not regularly smooth. It presented
small depression and was predominantly re-covered by fibrous tissue. The deep part presented rudimentarily neoformed trabecular bone. It
practically did not present formation of subchondral bone (Figure 4-B). Tabela 1 - Distribuição da freqüência de resultados na
avaliação macroscópica da cicatrização dos defeitos. Table 1 - Distribution of the frequency of results in the
macrocospic evaluation of defects healing. 1 - Aspecto macroscópico O típico aspecto macroscópi-
co encontrado nos defeitos trata-
dos e não tratados pode ser visto
nas Figuras 3-A e 4-A, resultados
considerados bons ou biologica-
mente aceitáveis. O termo “biolo-
gicamente aceitável”, segundo
Amiel(1) , é utilizado para definir re-
paros, que à observação macros-
cópica, se apresentam como te-
cido neo-formado semelhante à fi-
brocartilagem, liso, brilhante, firme, em con-
tinuidade com a cartilagem adjacente. Tra-
ta-se de um critério macroscópico de ava-
liação, não se podendo estabelecer uma
correlação exata com as características his-
tológicas, bioquímicas ou biomecânicas do
reparo. Figu
cicat
Figure
forma Figura 4 - Defeito não tratado: em “A” aspecto macroscópico; em “B”
cicatrização com formação de fibro cartilagem, superfície com pequena
depressão e osso subcondral desorganizado. Figure 4 - Untreated defect: in “A” macroscopic aspect; in “B” healing with
formation of fibrocartilage; surface with little depression and disorganized
subchondral bone.. Parâmetros
tratados
não tratados
Parâmeters
treated
untreated
n = 6
n = 6
n
%
n
%
Reparação do defeito
Reparação do defeito
Reparação do defeito
Reparação do defeito
Reparação do defeito
Defect repair
Defect repair
Defect repair
Defect repair
Defect repair
100%
20
100
19
95
75%
0
0
01
05
<75%>50%
0
0
00
00
<50%
0
0
00
00
Integração do defeito
Integração do defeito
Integração do defeito
Integração do defeito
Integração do defeito
Defect integration
Defect integration
Defect integration
Defect integration
Defect integration
100%
20
100
19
95
75%
0
0
01
05
<75%> 50%
0
0
00
00
<50%
0
0
00
00 In spite of presenting minimum alter-
ations of the surface these characteristics
were similar in the treated defects. Howev-
er, all the repairs of untreated defects or not
were considered satisfactory, therefore bio-
logically acceptable after twelve weeks of
evolution. Os dados relativos ao resultado da ava-
liação macroscópica, que incluiu os cinco
parâmetros já citados, estão resumidos nas
Tabelas 1 e 2. Observou-se predomínio da
formação de um tecido de reparação firme
com preenchimento total da lesão, com su-
perfície plana, regular e em continuidade
com a cartilagem adjacente nos defeitos
tratados. Nos defeitos não tratados essas
características foram semelhantes a não
ser por apresentarem alterações mínimas
da superfície. Porém todos os reparos pro-
venientes de defeitos tratados ou não, fo-
ram considerados satisfatórios, portanto bi-
ologicamente aceitáveis após doze sema-
nas de evolução. 2 - Aspecto histológico A reparação histológica foi considerada satisfatória somente se
a superfície do tecido regenerado fosse lisa e coberta por cartilagem
e ainda estando no mesmo nível da cartilagem adjacente. As diferen-
ças de tipo de tecido de cicatrização dos reparos foram nítidas. Para os defeitos tratados a superfície dos reparos mostraram a
preservação da cartilagem hialina original lisa, regular e no mesmo
nível da cartilagem adjacente hospedeira. A parte profunda do repa-
ro tinha se transformado em tecido ósseo com padrão trabecular
próximo da normalidade (Figura 3-B). 1 - Macroscopic aspect The typical macroscopic aspect in the treated and untreated de- 18 ACTA ORTOP BRAS 12(1) - JAN/MAR, 2004 Figura 3 - Defeito tratado: em “A” aspecto macroscópico; em “B”
cicatrização com formação de cartilagem hialina, superfície lisa e osso
subcondral normal. Figure 3 - Treated defect: in “A”: macroscopic aspect; in “B”: healing
with formation of hyaline cartilage; smooth surface and normal
subchondral bone fects can be seen in the Figures
3-A and 4-A; the results were con-
sidered as good or biologically
acceptable. “The biologically ac-
ceptable” term, according to
Amiel(1), is used to define repairs,
that in macroscopical observa-
tion, present as neo-formed tis-
sues similar to the fibrocartilage. They are smooth, brilliant, firm,
and in continuity with the adjacent
cartilage. As this is a macroscop-
ic criterion of evaluation, it is not
possible to settle an accurate cor-
relation among the histological,
biochemical or biomechanic
characteristics of the repair. The
relative data to the result of the
macroscopic evaluation, that in-
clude the five parameters already
mentioned, are summarized in
Tables 1 and 2. It was observed
the predominance of the forma-
tion of a firm repair tissue, with
total fulfilling of the injury, present-
ing a plain and regular surface in
continuity with the adjacent carti-
he treated defects. No exame histológico das sec-
ções do fêmur distal, a atenção foi
dirigida particularmente para os
seguintes elementos: 1- o tipo de
tecido regenerado na superfície
do defeito: cartilagem, fibrocarti-
lagem ou tecido fibroso; 2- o es-
tado da superfície cartilaginosa:
lisa, com depressão ou irregular;
3- presença ou ausência do osso
subcondral regenerado. Figura 3 - Defeito tratado: em “A” aspecto macroscópico; em “B”
cicatrização com formação de cartilagem hialina, superfície lisa e osso
subcondral normal. Figure 3 - Treated defect: in “A”: macroscopic aspect; in “B”: healing
with formation of hyaline cartilage; smooth surface and normal
subchondral bone. Figure 3 - Treated defect: in “A”: macroscopic aspect; in “B”: healing
with formation of hyaline cartilage; smooth surface and normal
subchondral bone. Figura 4 - Defeito não tratado: em “A” aspecto macroscópico; em “B”
cicatrização com formação de fibro cartilagem, superfície com pequena
depressão e osso subcondral desorganizado. Figure 4 - Untreated defect: in “A” macroscopic aspect; in “B” healing with
formation of fibrocartilage; surface with little depression and disorganized
subchondral bone
subchondral bone. ACTA ORTOP BRAS 12(1) - JAN/MAR, 2004 DISCUSSÃO Nem todas as lesões cartilaginosas são progressivas e nem to-
das são clinicamente sintomáticas, podendo a articulação retornar
ao seu estado normal(4,14) . Permanece a dúvida sobre qual lesão ne-
cessitaria de reparo cirúrgico e se o tratamento profilático alteraria
para melhor a evolução particular de cada lesão. The defects larger than 4 mm of diameter can practically take all
medial femoral condyle. The ones 5-mm height can cause destruction
to the bone marrow and such factors modify the sensitivity of the ex-
perimental model and the expected result(24). O coelho foi escolhido, neste estudo, por ser um animal ampla-
mente utilizado em modelos para reparação da superfície articular e
a forma cilíndrica do defeito por ser também unanimemente escolhi-
da quando se trata de estudo de cicatrização de lesões focais oste-
ocondrais(9,12,15,16,21,22). The amount of involved animals in this research was based on
bioethic principles. We used a minimum number for validation of hy-
pothesis(8,13). The six animals of the present series were selected from
a bigger group (n=20) of a wider study, exactly because all the repairs
were biologically acceptable independently if they were from treated
or untreated defects. All the knees of these 06 animals underwent mag-
netic resonance, in a previous study of a graduation thesis of the main
author(20). The examinations were performed at the end of the1st, 4th,
8th and 12th weeks of evolution with homogeneously good results in
what concerns the cicatrization, but not to the type of tissue formed on
the surface of the repairs. The aim of this present animal model was to
perform a basic histologic study, in a minimum sample that was homo-
geneous and known, in order to verify the type of tissue formed in both
types of repairs. Em relação ao tamanho da lesão optamos por criar defeitos de
3.2 mm/4.0 mm em animais acima de 3.5 kg de peso para se guar-
dar uma proporcionalidade adequada, pois defeito maiores de 4 mm
de diâmetro já tomam praticamente todo o côndilo femoral medial e
de 5 mm de altura já provocam destruição medular óssea e tais
fatores alteram a sensibilidade do modelo experimental e logica-
mente os resultados esperados(24). A quantidade de animais envolvidos nesta pesquisa baseou-se
em princípios bioéticos, buscando-se utilizar um número mínimo para
validação das hipóteses(8,13). DISCUSSION Not all the cartilaginous injuries are gradual and clinically symp-
tomatic. The articulation can return to its normal state(4,14). The uncer-
tainty about which injury should require surgical repair still remains and
it is not sure that the prophylactic treatment would modify for better the
particular evolution of each injury. A superfície dos reparos provenientes de defeitos não tratados
não era regularmente lisa, apresentava pequena depressão e estava 19 ACTA ORTOP BRAS 12(1) - JAN/MAR, 2004 Parâmetros
tratados
não tratados
Parameters
treated
untreated
n = 6
n = 6
n
%
n
%
Superfície / Surface
Surface
Surface
Surface
Surface
plana / smooth
20
200
17
85
depressão / depression
0
0
03
15
degeneração / degenerated
0
0
00
00
Coloração / Color
Color
Color
Color
Color
translúcida / translucide
19
95
19
95
opaca / opaque
01
05
01
05
descolorida / descolored
0
0
0
0
Erosão / Erosion
Erosion
Erosion
Erosion
Erosion
nenhuma / none
20
100
19
95
somente do defeito / only in the defect
0
0
01
05
defeito e cartilagem adjacente / defect and adjacent cartilage
0
0
0
0
Tabela 2 - Distribuição da freqüência de resultados na avaliação do aspecto macroscópico dos defeitos. Table 2 - Distribution of the frequency of results in the evaluation of the macrocospic aspect of the defects. The rabbit was chosen, in this study, because it is an animal widely
used in repairing models of the articular surface of the defect and the
cylindrical form can also be unanimously chosen if the case concerns
the study of cicatrization of osteochondral focal injuries(9,12,15,16,21,22). recoberta por tecido predominantemente fibroso. A parte profunda
apresentava osso trabecular neoformado rudimentarmente, não ha-
vendo praticamente formação de osso subcondral (Figura 4-B). recoberta por tecido predominantemente fibroso. A parte profunda
apresentava osso trabecular neoformado rudimentarmente, não ha-
vendo praticamente formação de osso subcondral (Figura 4-B). Regarding the size of the injury we decided to develop defects of
3.2 mm/4.0 mm in animals over 3.5 kg of weight in order to maintain an
adequate proportionality. ACTA ORTOP BRAS 12(1) - JAN/MAR, 2004 REFERÊNCIAS BIBLIOGRÁFICAS
REFERÊNCIAS BIBLIOGRÁFICAS
REFERÊNCIAS BIBLIOGRÁFICAS
REFERÊNCIAS BIBLIOGRÁFICAS
REFERÊNCIAS BIBLIOGRÁFICAS 1. Amiel D, Coutss RD, Abel M, Stewart W, Hailwood F, Akeson W H. Rib
perichondrial grafts for the repair of full thickness articular cartilage defects. A
morphological and biochemical study in rabbits. J Bone Joint Surg Am 67:911-
920, 1985. 1. Amiel D, Coutss RD, Abel M, Stewart W, Hailwood F, Akeson W H. Rib
perichondrial grafts for the repair of full thickness articular cartilage defects. A
morphological and biochemical study in rabbits. J Bone Joint Surg Am 67:911-
920, 1985. 2: Animal welfare requirements” Interpretação e extensão da norma. Acta Ortop
Bras 6:139-142, 1998. 14. Mankin HJ. Current concepts review. The response of articular cartilage to
mechanical injury. J Bone Joint Surg Am 64:460-466, 1982. 2. Brittberg M, Nilsson A, Lindahl A, Ohlsson C, Peterson L. Rabbit articular
cartilage defects treated with autologous cultured chondrocytes. Clin Orthop
326:270-283, 1996. 15. Mankin HJ. Localization of tritiated thymidine in articular cartilage of rabbits II:
Repair in immature cartilage. J Bone Joint Surg Am 44:688-698, 1962. 16. Meachin G, Roberts C. Repair of joint surface from subarticular tissue in the
rabbit knee. J Anat 109:317-327, 1971. 3. Calandruccio RA, Gilmer WS Jr. Proliferation, regeneration, and repair of articular
cartilage of immature animals. J Bone Joint Surg Am 44:431-455, 1962. 17. O’Driscoll SW, Keeley FW, Salter RB. The chondrogenic potential of free
autogenous periosteal grafts for biological resurfacing of major full-thickness
in joint surfaces under the influence of continuous passive motion. An
experimental investigation in the rabbit. J Bone Joint Surg Am 68:1017-1035,
1986. 4. Campbell CJ. The healing of cartilage deffects. Clin Orthop 64:45-63, 1969. 5. Convery FR, Akeson WH, Keown GH. The repair of large osteochondral defects:
an experimental study in horses. Clin Orthop 82:253-262, 1972. 6. De Palma AF, McKeever CD, Subin DK. Process of repair of articular cartilage
demonstrated by histology and autoradiography with tritiated thymidine. Clin
Orthop 48:229-242, 1966. 18. Peterson L, Minas T, Brittberg M, Nilsson A, Stogren-Jansson E, Lindahl A. Two to nine years outcome after autologous chondrocyte transplantation of
the knee. Clin Orthop 374:212-234, 2000. 7. Fuller JA, Ghadially FN. Ultrastructural observation on surgically produced
partial-thickness defectes in articular cartilage. Clin Orthop 86:193-205, 1972. 19. Pridie KH. A method of resurfacing osteoarthritis knee joints. In: Proceedings
of the British Orhopaedic Association. J Bone Joint Surg Br 41:618-619, 1959. 8. Gomes PF. Curso de estatística experimental. 2.ed. São Paulo: Universidade
de São Paulo, 1963. p.53-60. CONCLUSÕES Após doze semanas e nas condições experimentais descritas no
presente estudo pode-se concluir: 1 - Todos os defeitos osteocondrais tratados e não tratados evo-
luíram para reparos biologicamente aceitáveis 2 - A análise histológica dos reparos biologicamente aceitáveis
demonstrou formação de tecido cartilaginoso nos defeitos tratados
e de tecido fibrocartilaginoso nos defeitos não tratados. 2 - A análise histológica dos reparos biologicamente aceitáveis
demonstrou formação de tecido cartilaginoso nos defeitos tratados
e de tecido fibrocartilaginoso nos defeitos não tratados. CONCLUSIONS After twelve weeks under the described experimental conditions in
the present study, we can conclude: Reparos biologicamente aceitáveis podem se formar a partir de
defeitos tratados ou não e podem ser indistinguíveis pela aparência
macroscópica. O estudo histológico permitiu que se diferenciasse
ambos tipos de reparos. 1 - All the treated and untreated osteochondral defects became
biologically acceptable repairs. 2 - histologic analysis of the biologically acceptable repairs dem-
onstrated cartilaginous tissue and fibrocartilaginous tissue formation
in the treated or untreated defects respectively. Nas circunstâncias do estudo histológico ficou bem evidenciado
o tipo de tecido neoformado para cada tipo de reparo. A inferiorida-
de dos reparos de defeitos não tratados foi devida principalmente ao
retardo na formação de osso subcondral, o que mostra a fundamen-
tal importância do osso subcondral na reorganização tecidual. A pre-
visibilidade de formação de tecido cartilaginoso nos defeitos trata-
dos e de tecido predominantemente fibroso nos defeitos não trata-
dos estão de acordo com os dados da literatura(10,17,22). DISCUSSÃO Os seis animais da presente série foram
selecionados de um grupo maior (n=20) de um estudo mais amplo,
justamente por serem todos reparos biologicamente aceitáveis inde-
pendentemente se eram provindos de defeitos tratados ou não trata-
dos. Todos os joelhos desses 06 animais foram submetidos à resso-
nância magnética, em um estudo prévio de uma tese de doutorado
do autor principal(20). Os exames foram realizados ao final das 1ª, 4ª,
8ª e 12ª semanas de evolução com resultados homogeneamente
bons no que diz respeito à cicatrização, porem não no tipo de tecido
que se formava na superfície dos reparos. O que se pretendeu no
presente modelo animal foi realizar um estudo histológico básico,
em uma amostra mínima, homogênea e conhecida, somente para
averiguação do tipo de tecido que se formou em ambos tipos de
reparos. The result of 100% of biologically acceptable repairs according to
the settled macroscopic criteria, the absence of macroscopic alter-
ations in the articular tibia cartilage in contact with the femoral condyles,
the absence of infection and the maintenance of the articular stability
during the twelve weeks of evolution demonstrated the validity of the
experimental method applied. Biologically acceptable repairs can be
formed from treated defects or not and can be indistinguishable by the
macroscopic appearance. The histologic study allowed us to differen-
tiate both types of repairs. In the circumstances of the histologic study
it was well evidenced the type of neoformed tissue for each type of
repair. The inferiority of the repairs of treated defects was due to the
delay in the formation of the subchondral bone, what shows the basic
importance of the subchondral bone in the tissular reorganization. The
previsibility of the cartilaginous tissue formation in the treated defects 20 ACTA ORTOP BRAS 12(1) - JAN/MAR, 2004 and the fibrous tissue in the untreated defects is in accordance with
the literature data(10,17,22). and the fibrous tissue in the untreated defects is in accordance with
the literature data(10,17,22). A obtenção de 100% de reparos biologicamente aceitáveis, con-
forme os critérios macroscópicos estabelecidos, a ausência de alte-
rações macroscópicas na cartilagem articular tibial em contato com
os côndilos femorais, a ausência de infecção e a manutenção da
estabilidade articular durante as doze semanas de evolução demons-
tram a validade do método experimental empregado. REFERÊNCIAS BIBLIOGRÁFICAS
REFERÊNCIAS BIBLIOGRÁFICAS
REFERÊNCIAS BIBLIOGRÁFICAS
REFERÊNCIAS BIBLIOGRÁFICAS
REFERÊNCIAS BIBLIOGRÁFICAS 20. Ribeiro JL. Estudo comparativo da cicatrização de defeito osteocondral em
coelhos. São Paulo, 2003. Tese (Doutorado), Faculdade de Medicina da
Universidade de São Paulo. 9. Hangody L, Kish G, Karpati Z, Eberhart R. Osteochondral plugs: autogenous
osteochondral mosaicplasty for the treatment of focal chondral and
osteochondral articular defects. Oper Tech Orthop 7:312, 1997. 21. Rubak JM, Poussa M, Ritsilä V. Chondrogenesis in repair of articular cartilage
defects by free periosteal grafts in rabbits. Acta Orthop Scand 53:181-186,
1982. 10. Hangody L, Feczkó P, Bartha L, Bodó G, Kish G. Mosaicplasty for the treatment
of articular defects of the knee and ankle. Clin Orthop 391(Suppl):328-336,
2001. 22. Salter RB, Simmonds DF, Malcom BW, Rumble DJ, MacMichael D, Clements
ND. The biological effect of continuous passive motion on the healing of full
thickness defects on articular cartilage. An experimental investigation in the
rabbits. J Bone Joint Surg Am 62:1232-1251, 1980 11. Key J. Experimental arthritis: the changes in joints produced by creating defects
in the articular cartilage. J Bone Joint Surg Am 13:725, 1931. 12. Kim HK, Moran ME, Salter RB. The potential for regeneration of articular
cartilage in defects created by chondral shaving and subchondral abrasion. An experimental investigation in rabbits. J Bone Joint Surg Am 73:1301-1315,
1991. 23. Steadman JR, Rodkey WG, Briggs KK, Singleton SB. Microfracture technique
for full thickness chondral defects: technique and clinical results. Oper Tech
Orthop 7:300-304, 1997. 24. Wakitani S, Goto T, Pineda SJ, Young RG, Mansour JM, Caplan AJ, Golberg
VM. Mesenchymal cell-based repair of large full-thickness defects of articular
cartilage. J Bone Joint Surg Am 76:579-592, 1994. 13. Leopizzi N, Bollinger Neto R, Barros Filho TEP, Azze RJ. Modelos experimentais
em Ortopedia e ISSO 10993: “Biological evaluation of medical devices- Part 21 ACTA ORTOP BRAS 12(1) - JAN/MAR, 2004
|
https://openalex.org/W2010664424
|
https://europepmc.org/articles/PMC3885510?pdf=render
|
English
| null |
Impact of Universal Health Insurance Coverage on Hypertension Management: A Cross-National Study in the United States and England
|
PloS one
| 2,014
|
cc-by
| 8,894
|
Abstract This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits
tion, and reproduction in any medium, provided the original author and source are credited. Funding: There was no direct external funding for this study. The Department of Primary Care & Public Health at Imperial College is grateful for support from the
National Institute for Health Research Biomedical Research Centre scheme, the National Institute for Health Research Collaboration for Leadership in Applied
Health Research & Care scheme and the Imperial Centre for Patient Safety and Service Quality. Prof Wachter was a US-UK Fulbright Scholar at Imperial College
London in 2011. Dr Millett is funded by the Higher Education Funding Council for England and the National Institute for Health Research Collaboration for
Leadership in Applied Health Research & Care scheme. Dr Vamos is partly funded by the National Institute for Health Research. Dr Netuveli is supported by a grant
from the Economic & Social Research Council [RES-596-28-0001]. The funders had no part in the design and conduct of the study; collection, management,
analysis, and interpretation of the data; and preparation, review, or approval of the manuscript. Competing Interests: The authors have declared no competing interests exist. * E-mail: andrew.dalton@phc.ox.ac.uk ment, which are likely due to population, health care provider and
health system characteristics [4]. Andrew R. H. Dalton1*, Eszter P. Vamos2, Matthew J. Harris2, Gopalakrishnan Netuveli3,
Robert M. Wachter4, Azeem Majeed2, Christopher Millett2 1 Department of Primary Care Health Sciences, University of Oxford, Oxford, England, 2 Department of Primary Care and Public Health, Imperial College London, London,
England, 3 Institute for Health and Human Development, University of East London, London, England, 4 Division of Hospital Medicine, Department of Medicine, University
of California San Francisco, San Francisco, California, United States of America Abstract Background: The Patient Protection and Affordable Care Act (ACA) galvanised debate in the United States (US) over
universal health coverage. Comparison with countries providing universal coverage may illustrate whether the ACA can
improve health outcomes and reduce disparities. We aimed to compare quality and disparities in hypertension
management by socio-economic position in the US and England, the latter of which has universal health care. Method: We used data from the Health and Retirement Survey in the US, and the English Longitudinal Study for Aging from
England, including non-Hispanic White respondents aged 50–64 years (US market-based v NHS) and .65 years (US-
Medicare v NHS) with diagnosed hypertension. We compared blood pressure control to clinical guideline (140/90 mmHg)
and audit (150/90 mmHg) targets; mean systolic and diastolic blood pressure and antihypertensive prescribing, and
disparities in each by educational attainment, income and wealth, using regression models. Results: There were no significant differences in aggregate achievement of clinical targets aged 50 to 65 years (US market-
based vs. NHS- 62.3% vs. 61.3% [p = 0.835]). There was, however, greater control in the US in patients aged 65 years and over
(US Medicare vs. NHS- 53.5% vs. 58.2% [p = 0.043]). England had no significant socioeconomic disparity in blood pressure
control (60.9% vs. 63.5% [p = 0.588], high and low wealth aged $65 years). The US had socioeconomic differences in the 50–
64 years group (71.7% vs. 55.2% [p = 0.003], high and low wealth); these were attenuated but not abolished in Medicare
beneficiaries. Conclusion: Moves towards universal health coverage in the US may reduce disparities in hypertension management. The
current situation, providing universal coverage for residents aged 65 years and over, may not be sufficient for equality in
care. Citation: Dalton ARH, Vamos EP, Harris MJ, Netuveli G, Wachter RM, et al. (2014) Impact of Universal Health Insurance Coverage on Hypertension Management: A
Cross-National Study in the United States and England. PLoS ONE 9(1): e83705. doi:10.1371/journal.pone.0083705 Editor: Angelo Scuteri, INRCA, Italy Received June 26, 2013; Accepted November 6, 2013; Published January 8, 2014 Copyright: 2014 Dalton et al. This is an open-access article distributed under the terms of the Creative Commons Attributi
unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. ton et al. January 2014 | Volume 9 | Issue 1 | e83705 Impact of Universal Health Insurance Coverage on
Hypertension Management: A Cross-National Study in
the United States and England Andrew R. H. Dalton1*, Eszter P. Vamos2, Matthew J. Harris2, Gopalakrishnan Netuveli3,
Robert M. Wachter4, Azeem Majeed2, Christopher Millett2 Study sample We used data from Wave 4 of ELSA, conducted during 2008
and 2009, which contained 11,050 respondents (response rate
74.3%). We included respondents aged 50 years and over, with
valid blood pressure readings (the mean of two or more readings
on the same day_ these are considered valid for both surveys) from
the survey nurse visit (n = 7,982). To control for differences in
ethnic mix between samples, and ensure that we compared
differences between health systems – not biological differences in
control between samples, we restricted analyses to non-Hispanic
Whites (excluding 225 from ELSA). We exclude respondents with
missing SEP (n = 2,795) and clinical data (body mass index (BMI),
co-morbidities and smoking status; n = 900 – latter two not
mutually exclusive), leaving 4,910 respondents. Finally, England offers universal health coverage with care free
at the point of delivery. Medication in England costs a standard
prescription charge (£7.10 (,$13.90) in April 2008), although
exemptions mean that approximately 60 percent of the popula-
tion, and 90 percent of items dispensed are free of charge [11]. Exempt groups include people aged 60 years and over, people on
income support, or those with certain chronic health conditions –
although not hypertension. The US has universal coverage for
people aged 65 years and over, through Medicare. Co-payments
for medications, however, remain high, despite Medicare’s ‘‘Part
D’’ drug prescription benefit program [12]. Health coverage for
younger US patients is more variable, with a high proportion
without insurance or under-insured [4]. Having survived legal
contests over the ‘‘individual mandate’’ and given the outcome of
the
2012
presidential
election,
the
Patient
Protection
and
Affordable Care Act (ACA) will partly address this. It will expand
Medicaid eligibility and enhance individual access to private
health insurance, although the former may be limited by the
power of individual states not to implement changes [13]. For comparison we used data from Wave 9 of the HRS,
administered in 2008, with 17,217 respondents (response rate
88.6%). We include respondents aged 50 years and over, with
valid blood pressure readings (mean of two of more – n = 8,273). We excluded 3,049 non-white and Hispanic respondents, 304 with
no BMI record (there were no missing SEP data), leaving 4,920
respondents. From the samples defined above, we include respondents with a
self-reported physician’s diagnosis of hypertension, leaving 2,318
ELSA and 2,855 HRS respondents. Health Insurance Coverage and Hypertension Control in conditions requiring longitudinal care such as hypertension [9]. Second, as a single payer system, the NHS arguably has clearer
lines of accountability and more centralized coordination of
quality improvement (though the more pluralistic US system may
be more innovative). Thirdly, the US has considerably higher
health care spending than England; although this may be
considered a result of other structural differences [10]. In 2009
the US spent 17.4 percent of its Gross Domestic Product on health
care, compared with 9.8 percent in the United Kingdom (UK). This may provide resources for higher quality care, although the
relationship between health care spending and quality remains
unclear [10]. Outcome variables We analysed data from two national longitudinal surveys, the
English Longitudinal Study of Aging (ELSA), and the Health and
Retirement Survey (HRS) from the US. These provide compara-
ble data on aging, with similar sampling techniques and content
[5]. Both are population-based surveys presenting information on
health, physical and mental functioning, demographic factors,
behaviors and SEP in non-institutionalized individuals aged 50
years and over. We used mean systolic and diastolic blood pressure (from two or
more measures), defined controlled hypertension as blood pressure
lower than 140/90 mmHg, in line with clinical guidelines [17,18]
and measured prescribing of anti-hypertensive agents based on
self-report. In addition to the guideline-derived target, we assessed
control (in both countries) based on the UK’s audit target of 150/
90 mmHg. This defines control in England’s Quality and
Outcome Framework (QOF), which provides pay-for-performance
remuneration to primary care practices for the management of
chronic disease, including hypertension, therefore is an important
UK threshold for control [19]. ELSA, a biennial survey, began in 2002, sampling respondents
aged 50 years and over [15]. A refreshment sample, aged 50 to 53
years, was added in 2006 to boost representation of younger age
groups. Interviews are carried out at respondents’ homes, with
visiting nurses collecting biomedical data during alternate waves. The HRS started in 1991 [16], with biennial follow-up since 1992
and subsequent refreshment samples. The HRS has maintained a
sample of more than 20,000 respondents, and is conducted using
both telephone and face-to-face interviews – dependent on the
survey wave and module of questions. Further details of both
surveys can be found elsewhere [15,16]. Predictor variables – measures of SEP We used three measures of SEP that are comparable between
countries: education, household income and wealth. We divided
education into three groups defined using years of continuous
education; in England 0–11 years, 12–13 and $14; in the USA 0–
12, 13–15 and $16 years. These cut-offs are widely used [5,6],
producing similar education distributions across countries. We
used total household income, and total household wealth (savings,
net stock value, mutual funds, bonds, real estate value, own
business
share,
owned
cars
minus
liabilities),
adjusted
for
household size (the square root of household size), and producing
groups separated by tertiles within each country. Introduction Given the growing global burden of chronic diseases, effective
and equitable management of diseases such as hypertension is a
core requirement of any health system [1]. Despite population
level reductions in blood pressure and improvements in anti-
hypertensive prescribing [2,3],_ENREF_2 suboptimal hyperten-
sion management – most notably insufficient blood pressure
control – persists in many countries, including the United States
(US) and England. One marker of this quality gap across all
countries is within-country disparities in hypertension manage- Cross-country analysis can examine relationships between
health system structures and outcomes [5,6]. England and the
US have four important differences that may influence hyperten-
sion management. Firstly, the US has a more specialised health
service (higher ratio of specialists to generalists; more ambulatory
care delivered by specialists) whereas the National Health Service
(NHS) emphasizes generalist primary care, with restricted access to
specialists through gate-keeping. Strong primary care can improve
health outcomes [7,8] and may reduce inequalities [7], especially January 2014 | Volume 9 | Issue 1 | e83705 1 PLOS ONE | www.plosone.org January 2014 | Volume 9 | Issue 1 | e83705 Study sample Both surveys included a
question similar to ‘‘Has a doctor ever told you that you have
hypertension?’’ Though this method may underestimate hyperten-
sion rates compared with measures based on actual blood pressure
values [5], we felt it a more relevant definition for our study, which
relates to the management of patients with a known diagnosis –
further there is no evidence of inequalities in reporting-bias [15]. We aim to determine whether health system differences
between the US and England influence the quality of hypertension
management and disparities across socio-economic position (SEP),
characterised by income, wealth and education [14]. By stratifying
respondents into one group younger than 65 years and the other
older (i.e. the eligibility criteria for Medicare), we compare
hypertension management in the NHS both with part of the US
health care system with universal (Medicare) coverage and with
the market-based system for those under 65. The latter will see
coverage expanded as part of the ACA. US Market-based v English NHS (50–64 years) In patients with hypertension, we modelled blood pressure
control (to clinical and audit targets) and anti-hypertensive
medication using logistic regression [20]; and systolic and diastolic
blood pressure using linear regression. We modelled outcomes
separately in each country, age group and individual measure of
SEP; adjusting for age, sex, smoking status, vascular co-morbidity
and BMI. In the 50 to 64 years age group in the US, we present
results in the whole population and the insured group alone. We
defined the insured group as those reporting any health insurance
coverage (private, Medicaid or other health care plan) at the time
of interview. We present adjusted outcomes after recombination of
model coefficient [20], presenting the overal control in each
country and in each SEP group. We test statistical significance in
global control between countries; between SEP groups within each
country, comparing medium and high, with lowest group for each
indicator; and in equivalent SEP groups between countries. We
derived standard errors using the delta method for all adjusted
coefficiants [21]; when unable to estimate variance directly, this
allows accurate estimation [21]. We use valid survey weights in all
analyses (variables w4nurwt in ELSA and lwgtr in the HRS), which
we conducted using Stata 11.2SE. In patients with hypertension aged 50–64 years, there were no
significant differences in the control of blood pressure to 140/
90 mm Hg between countries (table 2). Respondents with health
insurance in the US had similar control to those in England (61.3
in England vs. 62.3 in all Americans (p = 0.835 vs. England) and
65.4% in the insured (p = 0.409 vs. England). Similarly there were
no statistically significant differences in control to audit targets in
the whole population (74.5 in England vs. 69.4 in the US;
p = 0.255), or insured Americans (72.7; p = 0.695). Despite no overall difference between countries in the 50–64
years age group, Americans in the highest wealth groups had
significantly better control to the clinical target than their English
counterparts (table 4) (71.7 vs. 60.9; p = 0.037). Wealth disparities
to both clinical and audit targets were present in the US, with
wealthier patients more likely to meet targets, but not in England
(clinical target- 60.9% for wealthy vs. 63.5% for poorer patients in
England (p = 0.588); compared with 71.7% vs. 55.2% (p = 0.003)
in the US). Health Insurance Coverage and Hypertension Control Health Insurance Coverage and Hypertension Control diagnoses out of coronary heart disease, stroke/transient ischaemic
attack, diabetes or chronic kidney disease. blood pressure control was present in highest SEP groups (e.g. income – lowest 53.9% in England vs. 56.3% in US, p = 0.533;
highest 51.0% vs. 62.9% respectively, p = 0.033). US Market-based v English NHS (50–64 years) The wealth disparity is reduced, but still significant in
the insured, under-65 American group (72.5 in the most wealthy
vs. 60.7 in the least (p = 0.036)). There was a suggestion of an
income disparity in the US sample, with the intermediate group
having better control than the lowest (clinical target – 54.9% in
low income vs. 66.6% in intermediate (p = 0.05)). This was not,
however statistically significant for the highest income (62.2%;
p = 0.198). There were no income disparities in the English
sample. US Medicare v English NHS (. 65 years) There was significantly higher aggregate blood pressure (BP)
control in US respondents aged .65 years using clinical
(BP,140/90 mm
Hg-
53.5%
England
vs. 58.2%
in
US;
p = 0.043) but not audit targets (BP,150/90 mm Hg- 71.9% vs. 73.9% respectively; p = 0.338) (table 2). Mean systolic BP was
significantly lower in the US than England (135.6 vs. 140.2 mm
Hg; p,0.001) but the converse for mean diastolic BP (78.9 vs. 73.7 mm Hg; p,0.001). Prescribing of at least one anti-
hypertensive medication was significantly more common in the
US than England (91.4% vs. 80.9%; p,0.001). Results The unadjusted prevalence of hypertension in England and the
US was 38.2% and 45.1% in respondents aged 50–64 years and
52.9% and 63.6% in those aged .65 years, respectively. There are
significant disparities in prevalence, especially by education and
wealth, which were greatest in US respondents aged 50–64 years. The characteristics of respondents with hypertension are presented
in table 1. In both age groups, there was higher mean BMI and
prevalence of vascular co-morbidity in the US. There are marked
gradients in smoking in both countries, with a higher prevalence in
lowest SEP groups and, in younger patients, higher BMI in lowest
SEP groups. There are significant SEP disparities in insurance
coverage in US respondents aged 50–64 years. Mean systolic blood pressure was lower in the total and insured
American groups aged under 65 years compared with the sample
from
England
(England = 135.5;
insured
American = 129.9
(p,0.001 vs. England); total American = 131.2 (p = 0.007 vs. England)): Diastolic blood pressure was lower in the English
sample, compared with both from the US (England = 81.5; insured
American = 83.9 (p,0.016 vs. England); total American = 84.8
(p,0.001 vs. England)). English patients with hypertension were less often prescribed
anti-hypertensive medication than insured Americans and Amer-
icans in total (56.4%, 84.2% (p,0.001 vs. England) and 80.2%
(p,0.001
vs. England),
respectively). There
were
no
SEP
disparities in prescribing in the US Sample, but there was
statistically significantly lower prescribing in the lowest wealth
group in the total US sample compared with the lowest (71.2% in
lowest vs. 86.0% in the highest; p = 0.004). There were no
prescribing
disparities
in
the
US
sample
reporting
health
insurance. Statistical Methods There was higher prescribing of anti-hypertensive medication in
the US across all SEP groups compared with England. In England
there was no SEP disparity in prescribing, whereas the US had
lower prescribing in lowest SEP groups, which was statistically
significant for wealth (87.4% in the least wealthy vs. 92.4% in the
most; p = 0.029). We conducted analyses separately in the groups aged 50 to 64
years and 65 years and over. We present the prevalence of
hypertension, standardized for age and sex, along with summaries
of covariates across indicators of SEP (education, income and
wealth), in patients with hypertension in each country, testing for
statistical significance. US Market-based v English NHS (50–64 years) Covariates We
controlled
for
the
following
potentially
confounding
variables: age and sex of respondents; smoking status (current/
non-smoker); BMI; and concordant vascular co-morbidities. All
except BMI were self-reported, which was recorded by study
nurses. We defined the latter as the sum of patient reported doctor January 2014 | Volume 9 | Issue 1 | e83705 January 2014 | Volume 9 | Issue 1 | e83705 PLOS ONE | www.plosone.org 2 Main findings The lowest SEP groups had poorer control to clinical targets in
the US, which reached statistical significance for income (56.3% in
low income vs. 62.9% in high; p = 0.033), but not in England
(table 3). There were no statistically significant SEP disparities to
audit targets. When the analysis compared equivalent SEP groups
between countries (results not presented), the largest disparity in Patients with hypertension in America eligible for Medicare are,
overall, more likely than their English counterparts to have their
blood pressure controlled to the clinical guideline target, but not to
the English audit target. Patients aged 50 to 64 years, with
insurance coverage through the NHS or under the market-based January 2014 | Volume 9 | Issue 1 | e83705 January 2014 | Volume 9 | Issue 1 | e83705 PLOS ONE | www.plosone.org 3 Health Insurance Coverage and Hypertension Control nd characteristics in hypertensive patients aged greater than 50 years in England and the United States. Table 1. Risk factors and characteristics in hypertensive patients aged greater than 50 years in England and the United States. Table 1. Risk factors and characteristics in hypertensive patients aged greater than 50 years in England and the United States. Main findings England
United States
Years of education
Low
Medium
High
Total
Low
Medium
High
Total
Age 50 to 64 years
N hypertensive¤
451
63
212
726
340
164
164
668
Prevalence, % (SE)
35.9 (2.2)
30.2 (3.5)
30.1 (2.2)
33.4 (1.8)
47.3 (2.4)`
43 (2.9)`
33.4 (2.6)
41.7 (1.7)`
Sex (% Female, SE)¤
54.0 (2.4)
39.9 (6.1)
40.0 (3.3)
49.3 (1.9)
48.5 (3.8)
38.1 (5.1)
35.6 (5.3)
42.8 (2.7)
Age, mean (SE)¤
60.3 (0.1)
59.7 (0.3)
59.9 (0.2)
60.1 (0.1)
59.2 (0.2)
59.1 (0.3)
59.2 (0.3)
59.2 (0.2)
Current smoker, % (SE)
20.5 (2.0)
12.6 (4.4)
8.2 (2.2)
16.7 (1.5)
25.0 (2.7)
19.3 (3.5)
11.6 (3.0)
20.0 (1.8)
BMI mean (SE)
30.6 (0.3)
29.9 (0.7)
29.3 (0.3)
30.2 (0.2)
31.5 (0.5)
31.9 (0.7)`
30.7 (0.7)
31.4 (0.3)`
.1 Vascular Co Morb, % (SE)
17.3 (1.8)
20.5 (5.2)
12.1 (2.2)
16.2 (1.4)
38.4 (3.7)`
27.9 (4.9)`
20.4 (4.8)`
31.4 (2.5)`
Insurance coverage, % (SE)
86.0 (2.6)
94.2 (2.4)
96.0 (2.5)
90.5 (1.6)
Age$65 years
N hypertensive¤
1195
68
329
1592
1344
416
427
2187
Prevalence, % (SE)
55.0 (1.1)
46.1 (4.1)
48.4 (1.9)
53.4 (0.9)
65.2 (1.2)`
61.7 (2.1)`
58.9 (2.0)`
63.1 (0.9)`
Sex (% Female, SE)¤
61.3 (1.4)
43.3 (6.0)
39.3 (2.7)
57.2 (1.3)
54.3 (1.9)
51.6 (3.4)
33.3 (3.1)
49.7 (1.5)
Age, mean (SE)¤
76.6 (0.2)
75.3 (1.0)
74.3 (0. 5)
76.2 (0.2)
74.8 (0.3)
73.7 (0.5)
74.0 (0. January 2014 | Volume 9 | Issue 1 | e83705 PLOS ONE | www.plosone.org
4
January 2014 | Volume 9 | Issue 1 | e83705 Main findings US health system, have similar control to both targets, even
limiting the comparison to US patients reporting health insurance. The US has significantly more prescribing of anti-hypertensive
medication across all levels of SEP. We found no evidence of
disparities in English patients, while significant wealth and income
disparities in blood pressure control and prescribing among
younger US patients. Disparities in both were generally attenuated
but not abolished in insured patients aged under-65 and older
patients eligible for Medicare. The US is enacting some of its most significant health care
reforms since the formation of Medicare in 1965 [22]. Currently,
up to 46 million Americans are uninsured [22], with gaps in health
insurance estimated to account for up to 45,000 deaths per year
[23]. The ACA aims to extend health insurance coverage to an
additional 31 million citizens. An open question is whether access
to health insurance will improve disease management and reduce
health disparities. In addition to more equitable care, the NHS matches the US in
absolute levels of blood pressure control in the sample aged 50 to
64 years. Given greater health care spending in the US [10], this
may suggest more efficient hypertension management under the
NHS than in a US system with no mandated health insurance
Reasons for this may include increased access to primary care, less
overcharging, less duplication of care, less overuse and lower
transaction/overhead costs [10,30]. Moreover, in the US system,
patients – even those with health insurance – are considerably
more likely to forgo necessary care due to cost and less likely to
have a long-term relationship with a health care provider [27]. Patients in the US eligible for Medicare do, however, see better
control, than the sample aged $65 years under the NHS. This
supports previous findings of greater blood pressure control in
patients with hypertension in the US than in England [31]. The
US may have improved treatment strategies, including the
medication use seen here. Systolic blood pressure was lower in
the US sample, but diastolic blood pressure significantly lower in
England (up to a 5 mm. Hg. difference aged $65). Diastolic blood
pressure may be more easily controlled with medication than
systolic blood pressure [32]. While health insurance may reduce health disparities through
improved access to care, the US and UK health systems differ in
other ways. Main findings 5)
74.4 (0.2)
Current smoker, % (SE)
10.3 (0.9)
4.7 (2.7)
5.2 (1.2)
9.3 (0.8)
11.1 (0.9)
9.6 (1.8)
5.0 (1.2)
9.6 (0.7)
BMI mean (SE)
28.9 (0.2)
29.2 (0.8)
28.2 (0.3)
28.8 (0.1)
29.8 (0.2)`
29.8 (0.5)
29.2 (0.3)`
29.7 (0.2)`
.1 Vascular Co Morb, % (SE)
27.5 (1.3)
33.4 (5.7)
24.9 (2.4)
27.3 (1.1)
41.1 (1.9)`
35.0 (3.2)
31.6 (3.1)`
38.0 (1.4)`
Income
Low
Medium
High
Total
Low
Medium
High
Total
Age 50 to 64 years
N hypertensive¤
184
250
292
726
161
196
311
668
Prevalence, % (SE)
37.7 (2.8)
35.2 (2.5)
30.1 (2.0)
33.4 (1.8)
47.8 (3.4)`
45.2 (3.0)`
38.1 (2.1)`
41.7 (1.7)`
Sex (% Female, SE)¤
51.6 (3.8)
49.7 (3.2)
47.2 (2.9)
49.3 (1.9)
53.1 (5.5)
47.1 (5.1)
34.5 (3.7)
42.8 (2.7)
Age, mean (SE)¤
60.1 (0.2)
60.4 (0.2)
60.0 (0.2)
60.1 (0.1)
59.0 (0.3)
59.2 (0.3)
59.3 (0.2)
59.2 (0.2)
Current smoker, % (SE)
23.7 (3.3)
18.4 (2.6)
10.1 (1.9)
16.7 (1.5)
29.0 (4.1)
24.0 (3.5)
13.6 (2.2)
20.0 (1.8)
BMI mean (SE)
31.2 (0.5)
30.2 (0.4)
29.6 (0.3)
30.2 (0.2)
32.3 (0.7)
31.6 (0.7)`
30.9 (0.5)
31.4 (0.3)`
.1 Vascular Co Morb, % (SE)
23.2 (3.2)
16.9 (2.4)
10.5 (1.8)
16.2 (1.4)
53.3 (5.4)`
24.0 (4.4)`
23.7 (3.4)`
31.4 (2.5)`
Insurance coverage, % (SE)
75.3 (4.7)
94.0 (2.2)
96.8 (1.4)
90.5 (1.6)
Age$65 years
N hypertensive¤
626
633
333
1592
762
856
569
2187
Prevalence, % (SE)
54.8 (1.5)
54.3 (1.5)
49.0 (1.9)
53.4 (0.9)
68.5 (1.6)`
63.2 (1.4)`
57.2 (1.7)`
63.1 (0.9)`
Sex (% Female, SE)¤
64.8 (1.9)
53.6 (2.0)
47.1 (2.8)
57.2 (1.3)
55.4 (2.6)
50.3 (2.3)
41.6 (2.8)
49.7 (1.5)
Age, mean (SE)¤
77.5 (0.3)
75.9 (0.3)
74.0 (0.4)
76.2 (0.2)
74.9 (0.4)
74.3 (0.3)
73.9 (0.4)
74.4 (0.2)
Current smoker, % (SE)
10.1 (1.2)
9.9 (1.2)
6.0 (1.3)
9.3 (0.8)
12.9 (1.4)
9.5 (1.1)
5.3 (1.0)
9.6 (0.7)
BMI mean (SE)
28.6 (0.2)
28.9 (0.2)
29.0 (0.3)
28.8 (0.1)
29.1 (0.3)`
30.1 (0.3)`
29.6 (0.3)
29.7 (0.2)`
.1 Vascular Co Morb, % (SE)
26.7 (1.8)
28.5 (1.8)
26.1 (2.5)
27.3 (1.1)
47.4 (2.6)`
36.0 (2.2)`
29.4 (2.6)
38.0 (1.4)`
Wealth
Low
Medium
High
Total
Low
Medium
High
Total
Age 50 to 64 years
N hypertensive¤
243
224
259
726
237
225
206
668
Prevalence, % (SE)
43.3 (2.7)
31.0 (2.3)
28.1 (2.1)
33.4 (1.8)
53.1 (2.9)`
42.2 (2.6)`
32.1 (2.4)
41.7 (1.7)`
Sex (% Female, SE)¤
48.2 (3.2)
53.8 (3.4)
46.2 (3.2)
49.3 (1.9)
48.1 (4.5)
41.7 (4.7)
37.2 (4.6)
42.8 (2.7)
Age, mean (SE)¤
60.1 (0.2)
60.2 (0.2)
60.1 (0.2)
60.1 (0.1)
58.9 (0.3)
59.1 (0.3)
59.6 (0.3)
59.2 (0.2)
Current smoker, % (SE)
27.3 (2.9)
13.9 (2.4)
7.5 (1.8)
16.7 (1.5)
30.0 (3.4)
18.8 (3.0)
10.1 (2.4)
20.0 (1.8)
BMI mean (SE)
31.4 (0.4)
30.0 (0.3)
29.2 (0.3)
30.2 (0.2)
32.1 (0.6)
31.8 (0.5)
30.3 (0.5)
31.4 (0.3)`
.1 Vascular Co Morb, % (SE)
24.4 (2.8)
13.1 (2.2)
10.0 (1.9)
16.2 (1.4)
44.9 (4.4)`
23.6 (4.1)`
22.6 (4.2)`
31.4 (2.5)`
Insurance coverage, % (SE)
83.5 (3.3)
91.9 (2.6)
97.7 (1.6)
90.5 (1.6) PLOS ONE | www.plosone.org 4 Health Insurance Coverage and Hypertension Control Table 1. Main findings Cont. England
United States
Years of education
Low
Medium
High
Total
Low
Medium
High
Total
Age$65 years
N hypertensive¤
513
594
485
1592
586
778
823
2187
Prevalence, % (SE)
58.4 (1.7)
54.1 (1.5)
47.4 (1.6)
53.4 (0.9)
67.8 (1.8)`
65.2 (1.5)`
58.7 (1.4)`
63.1 (0.9)`
Sex (% Female, SE)¤
62.6 (2.2)
57.1 (2.0)
50.0 (2.3)
57.2 (1.3)
61.1 (2.8)
47.6 (2.5)
43.4 (2.3)
49.7 (1.5)
Age, mean (SE)¤
77.2 (0.4)
76.1 (0.4)
75.1 (0.4)
76.2 (0.2)
74.5 (0.4)
74.3 (0.4)
74.4 (0.3)
74.4 (0.2)
Current smoker, % (SE)
15.3 (1.6)
6.5 (1.0)
5.1 (1.1)
9.3 (0.8)
14.8 (1.6)
9.8 (1.2)
5.7 (1.0)
9.6 (0.7)
BMI mean (SE)
29.1 (0.2)
29.0 (0.2)
28.0 (0.2)
28.8 (0.1)
29.4 (0.4)
29.9 (0.3)`
29.6 (0.3)`
29.7 (0.2)`
.1 Vascular Co Morb, % (SE)
34.1 (2.1)
26.0 (1.8)
19.8 (1.9)
27.3 (1.1)
47.2 (2.9)`
38.7 (2.5)`
30.9 (2.2)`
38.0 (1.4)`
BMI = Body Mass Index; BP = Blood Pressure; SE = standard error; Co Morb = co-morbidity; Age controlled to 57 in former age group and 75 in latter. ¤Not tested for statistical significance controlling for age and sex. {p,0.05. `p,0.01 testing the English against the US, controlling for age and sex. doi:10.1371/journal.pone.0083705.t001 Table 1. Cont. patients with chronic conditions in the US report significant out-
of-pocket payments (.$1,000), compared with only 4 percent in
the UK [27]. Moves under the ACA not only to extend health
insurance but also to impose minimum quality standards on all
policies, including ending annual claims limits and eliminating
certain co-payments, may limit obstacles to appropriate care for
Americans,
and
improve
equity
in
care
[13]. Alternately,
inequalities formed under the age of 65 years may simply become
too entrenched to be fully eradicated under Medicare. SEP may
be a fundamental cause of raised blood pressure [28]; however, health
systems should still seek to provide equitable management for
those with diagnosed hypertension [29]. US health system, have similar control to both targets, even
limiting the comparison to US patients reporting health insurance. The US has significantly more prescribing of anti-hypertensive
medication across all levels of SEP. We found no evidence of
disparities in English patients, while significant wealth and income
disparities in blood pressure control and prescribing among
younger US patients. Disparities in both were generally attenuated
but not abolished in insured patients aged under-65 and older
patients eligible for Medicare. January 2014 | Volume 9 | Issue 1 | e83705 Main findings England
United States
Total
Total
Insured
Aged $65 yearsa
Systolic BP mean (SE)
140.2 (0.65)
135.6 (0.64)`
–
Diastolic BP mean (SE)
73.7 (0.36)
78.9 (0.34)`
–
BP ,140/90 mmHg, % (SE)
53.5 (1.73)
58.2 (1.54){
–
BP ,150/90 mmHg, % (SE)
71.9 (1.55)
73.9 (1.39)
–
Anti-hypertensive medication, % (SE)
80.9 (1.34)
91.4 (0.91)`
–
Aged 50-64 yearsb
Systolic BP mean (SE)
135.5 (1.17)
131.2 (1.05)`
129.9 (1.09)`
Diastolic BP mean (SE)
81.5 (0.70)
84.8 (0.68)`
83.9 (0.68){
BP ,140/90 mmHg, % (SE)
61.3 (3.55)
62.3 (3.15)
65.4 (3.33)
BP ,150/90 mmHg, % (SE)
74.5 (3.24)
69.4 (3.06)
72.7 (3.15)
Anti-hypertensive medication, % (SE)
56.4 (3.71)
80.2 (2.61)`
84.2 (2.56)` Table 2. Aggregate blood pressure control and prescribing in hypertensive patients aged 50- 64 in the United States and
England. Table 3. Blood pressure control and prescribing in hypertensive patients in US Medicare v English NHS by socioeconomic-position
(. 65 years) Table 3. Blood pressure control and prescribing in hypertensive patients in US Medicare v English NHS by socioeconomic-position
(. 65 years). Main findings England
United States
Years of Education
Low
Medium
High
Low
Medium
High
Systolic BP mean (SE)
140.1 (0.73)
142.1 (3.00)
140.0 (1.11)
136.7 (0.78)
133.9 (1.24){
134.6 (1.05)
Diastolic BP mean (SE)
73.4 (0.41)
75.6 (1.44)
74.8 (0.64){
79.0 (0.42)
78.3 (0.65)
79.4 (0.57)
BP ,140/90 mmHg, % (SE)
53.6 (1.94)
47.8 (6.30)
54.2 (3.03)
56.8 (1.90)
57.9 (2.83)
61.5 (2.67)
BP ,150/90 mmHg, % (SE)
71.8 (1.76)
66.6 (6.20)
73.1 (2.79)
72.8 (1.71)
73.6 (2.51)
76.5 (2.35)
Anti-hypertensive medication, % (SE)
81.0 (1.48)
80.0 (5.16)
80.3 (2.45)
90.3 (1.21)
92.7 (1.87)
92.8 (1.54)
Income
Low
Medium
High
Low
Medium
High
Systolic BP mean (SE)
139.9 (0.96)
139.9 (0.89)
141.1 (1.16)
136.8 (1.10)
136.3 (0.86)
133.7 (0.94){
Diastolic BP mean (SE)
73.4 (0.53)
73.7 (0.51)
74.4 (0.63)
79.6 (0.58)
78.7 (0.47)
78.6 (0.50)
BP ,140/90 mmHg, % (SE)
53.9 (2.49)
54.5 (2.37)
51.0 (3.04)
56.3 (2.43)
55.8 (2.12)
62.9 (2.29){
BP ,150/90 mmHg, % (SE)
72.0 (2.27)
74.3 (2.11)
67.2 (2.89)
72.4 (2.20)
71.9 (1.95)
77.5 (1.96)
Anti-hypertensive medication, % (SE)
81.7 (1.88)
80.5 (1.80)
79.9 (2.48)
89.6 (1.88)
92.7 (1.10)
91.4 (1.46)
Wealth
Low
Medium
High
Low
Medium
High
Systolic BP mean (SE)
141.1 (1.10)
139.4 (0.94)
140.3 (0.93)
135.4 (1.21)
137.0 (0.93)
134.7 (0.82)
Diastolic BP mean (SE)
73.1 (0.62)
73.7 (0.52)
74.3 (0.52)
79.1 (0.65)
79.1 (0.50)
78.8 (0.45)
BP ,140/90 mmHg, % (SE)
51.2 (2.87)
55.4 (2.41)
53.1 (2.50)
56.6 (2.76)
56.3 (2.21)
60.3 (1.99)
BP ,150/90 mmHg, % (SE)
69.5 (2.70)
74.8 (2.15)
70.3 (2.31)
74.2 (2.46)
71.1 (2.02)
75.7 (1.74)
Anti-hypertensive medication, % (SE)
80.6 (2.16)
80.0 (1.90)
82.0 (1.96)
87.4 (2.46)
92.4 (1.23){
92.4 (1.11){
BP = Blood Pressure; SE = standard error; Values and significance testing controls for age and sex, body mass index, vascular co-morbidity and smoking (age controlled at
75, female sex, non-smoker, no co-morbidity and BMI = 28.8 – the English mean). Significance tests compare the medium and high SEP groups with the low within each
country. g
yp Table 2. Aggregate blood pressure control and prescribing in hypertensive patients aged 50- 64 in the United States and
England. Main findings Primary care plays a significantly greater role in
England, which itself may reduce disparities [7,24]. Importantly,
we not only saw reduced disparities in England compared with the
US, but also in American patients eligible for Medicare compared
with younger patients in the US. Insurance coverage may be a
significant driver of disparities in hypertension control in younger
Americans. In the US, younger patients with market-based (often
private) insurance may trade insurance source (switching to the
Medicare) once eligible. Younger patients lacking insurance
(usually from a lower SEP) are more likely to become newly
eligible for insurance at the age of 65 years [25] and in turn see
greatest increases in health care expenditure [26]. This might, in
part, reduce US disparities in hypertension management. That said, disparities in blood pressure control are absent in
older English patients but some remain in Medicare-eligible
Americans (for example an income disparity to clinical blood
pressure targets). Universal coverage alone may not eliminate
disparities. The strength of English primary care may increase
equity in hypertension control [7]. Also, in the US, even Medicare
patients, and many with private insurance, face significant co-
payments. These may stop patients obtaining necessary care,
including office visits and pharmaceuticals. Up to 40 percent of One interesting finding is that anti-hypertensive medications are
more far more frequently prescribed in the US sample aged 50 to
64 years, but control remains comparable. The control of blood
pressure in hypertensive patients is not solely determined by the
prescribing of pharmacological agents per se. Other factors,
including use of appropriate combination therapy and life-style
interventions, are important [33]. This might, at least in part,
explain the lack of association between prescribing rates and blood January 2014 | Volume 9 | Issue 1 | e83705 January 2014 | Volume 9 | Issue 1 | e83705 PLOS ONE | www.plosone.org 5 Health Insurance Coverage and Hypertension Control Table 2. Aggregate blood pressure control and prescribing in hypertensive patients aged 50- 64 in the United States and
England. Main findings England
United States
Total
Total
Insured
Aged $65 yearsa
Systolic BP mean (SE)
140.2 (0.65)
135.6 (0.64)`
–
Diastolic BP mean (SE)
73.7 (0.36)
78.9 (0.34)`
–
BP ,140/90 mmHg, % (SE)
53.5 (1.73)
58.2 (1.54){
–
BP ,150/90 mmHg, % (SE)
71.9 (1.55)
73.9 (1.39)
–
Anti-hypertensive medication, % (SE)
80.9 (1.34)
91.4 (0.91)`
–
Aged 50-64 yearsb
Systolic BP mean (SE)
135.5 (1.17)
131.2 (1.05)`
129.9 (1.09)`
Diastolic BP mean (SE)
81.5 (0.70)
84.8 (0.68)`
83.9 (0.68){
BP ,140/90 mmHg, % (SE)
61.3 (3.55)
62.3 (3.15)
65.4 (3.33)
BP ,150/90 mmHg, % (SE)
74.5 (3.24)
69.4 (3.06)
72.7 (3.15)
Anti-hypertensive medication, % (SE)
56.4 (3.71)
80.2 (2.61)`
84.2 (2.56)`
{p,0.05. `p,0.01 testing US groups against the English total. aUS Medicare v English NHS group. bUS market-based v English NHS; BP = Blood Pressure; SE = standard error. All values and significance testing controls for age and sex, BMI, vascular co-morbidity and smoking (age controlled at 75 & 57 respectively, female sex, non-smoker, no
co-morbidity and BMI = 28.8 – the English mean). doi:10.1371/journal.pone.0083705.t002
Table 3. Blood pressure control and prescribing in hypertensive patients in US Medicare v English NHS by socioeconomic-position
(. 65 years). Main findings England
United States
Years of Education
Low
Medium
High
Low
Medium
High
Systolic BP mean (SE)
140.1 (0.73)
142.1 (3.00)
140.0 (1.11)
136.7 (0.78)
133.9 (1.24){
134.6 (1.05)
Diastolic BP mean (SE)
73.4 (0.41)
75.6 (1.44)
74.8 (0.64){
79.0 (0.42)
78.3 (0.65)
79.4 (0.57)
BP ,140/90 mmHg, % (SE)
53.6 (1.94)
47.8 (6.30)
54.2 (3.03)
56.8 (1.90)
57.9 (2.83)
61.5 (2.67)
BP ,150/90 mmHg, % (SE)
71.8 (1.76)
66.6 (6.20)
73.1 (2.79)
72.8 (1.71)
73.6 (2.51)
76.5 (2.35)
Anti-hypertensive medication, % (SE)
81.0 (1.48)
80.0 (5.16)
80.3 (2.45)
90.3 (1.21)
92.7 (1.87)
92.8 (1.54)
Income
Low
Medium
High
Low
Medium
High
Systolic BP mean (SE)
139.9 (0.96)
139.9 (0.89)
141.1 (1.16)
136.8 (1.10)
136.3 (0.86)
133.7 (0.94){
Diastolic BP mean (SE)
73.4 (0.53)
73.7 (0.51)
74.4 (0.63)
79.6 (0.58)
78.7 (0.47)
78.6 (0.50)
BP ,140/90 mmHg, % (SE)
53.9 (2.49)
54.5 (2.37)
51.0 (3.04)
56.3 (2.43)
55.8 (2.12)
62.9 (2.29){
BP ,150/90 mmHg, % (SE)
72.0 (2.27)
74.3 (2.11)
67.2 (2.89)
72.4 (2.20)
71.9 (1.95)
77.5 (1.96)
Anti-hypertensive medication, % (SE)
81.7 (1.88)
80.5 (1.80)
79.9 (2.48)
89.6 (1.88)
92.7 (1.10)
91.4 (1.46)
Wealth
Low
Medium
High
Low
Medium
High
Systolic BP mean (SE)
141.1 (1.10)
139.4 (0.94)
140.3 (0.93)
135.4 (1.21)
137.0 (0.93)
134.7 (0.82)
Diastolic BP mean (SE)
73.1 (0.62)
73.7 (0.52)
74.3 (0.52)
79.1 (0.65)
79.1 (0.50)
78.8 (0.45)
BP ,140/90 mmHg, % (SE)
51.2 (2.87)
55.4 (2.41)
53.1 (2.50)
56.6 (2.76)
56.3 (2.21)
60.3 (1.99)
BP ,150/90 mmHg, % (SE)
69.5 (2.70)
74.8 (2.15)
70.3 (2.31)
74.2 (2.46)
71.1 (2.02)
75.7 (1.74)
Anti-hypertensive medication, % (SE)
80.6 (2.16)
80.0 (1.90)
82.0 (1.96)
87.4 (2.46)
92.4 (1.23){
92.4 (1.11){
BP = Blood Pressure; SE = standard error; Values and significance testing controls for age and sex, body mass index, vascular co-morbidity and smoking (age controlled at
75, female sex, non-smoker, no co-morbidity and BMI = 28.8 – the English mean). Significance tests compare the medium and high SEP groups with the low within each
country. {p,0.05. doi:10.1371/journal.pone.0083705.t003 BP = Blood Pressure; SE = standard error; Values and significance testing controls for age and sex, body mass index, vascular co-morbidity and smoking (age controlled at
75, female sex, non-smoker, no co-morbidity and BMI = 28.8 – the English mean). Significance tests compare the medium and high SEP groups with the low within each
country. {p,0.05. Main findings doi:10 1371/journal pone 0083705 t003 January 2014 | Volume 9 | Issue 1 | e83705 PLOS ONE | www.plosone.org Health Insurance Coverage and Hypertension Control Table 4. Blood pressure control and prescribing in hypertensive patients in US market-based v English NHS by socioeconomic-
position (aged 50-64 years). Main findings England
United States
Total
Total
Insured
Years of Education
Low
Medium
High
Low
Medium
High
Low
Medium
High
Systolic BP mean (SE)
136.9 (1.35) 131.3 (2.10)` 134.2 (1.46)
132.8 (1.42)
131.5 (1.70)
129.0 (1.45){
131.2 (1.50) 130.6 (1.76) 128.0 (1.46)
Diastolic BP mean (SE)
81.9 (0.82)
81.0 (1.47)
81.1 (0.85)
84.5 (0.93)
85.0 (1.04)
84.9 (0.95)
83.4 (0.98)
84.3 (1.00)
84.0 (0.93)
BP ,140/90 mmHg, % (SE)
57.8 (4.17)
70.3 (6.52)
64.6 (4.30)
59.3 (4.18)
59.7 (4.80)
68.0 (4.20)
63.2 (4.46)
62.5 (5.00)
70.1 (4.28)
BP ,150/90 mmHg, % (SE)
71.9 (3.89)
76.6 (6.15)
78.2 (3.71)
67.4 (4.09)
70.2 (4.61)
70.9 (4.21)
71.7 (4.22)
72.4 (4.64)
73.8 (4.22)
Anti-hypertensive medication, % (SE)
53.4 (4.32)
58.0 (7.06)
61.3 (4.58)
78.4 (3.89)
77.4 (4.23)
84.4 (3.50)
84.3 (3.87)
80.2 (4.01)
87.2 (3.32)
Income
Low
Medium
High
Low
Medium
High
Low
Medium
High
Systolic BP mean (SE)
134.0 (1.62) 136.3 (1.61) 135.7 (1.35)
135.5 (1.87)
129.5 (1.63)`
130.9 (1.22){
132.6 (2.09) 128.2 (1.70) 130.2 (1.24)
Diastolic BP mean (SE)
81.4 (0.97)
81.7 (0.97)
81.5 (0.82)
86.5 (1.18)
83.3 (1.09){
85.1 (0.75)
84.4 (1.32)
82.3 (1.13)
84.4 (0.73)
BP ,140/90 mmHg, % (SE)
62.6 (4.95)
58.1 (4.77)
62.6 (4.01)
54.9 (5.67)
66.6 (4.42){
62.2 (3.60)
62.4 (6.38)
69.1 (4.58)
64.5 (3.73)
BP ,150/90 mmHg, % (SE)
75.3 (4.52)
74.2 (4.24)
74.2 (3.68)
62.5 (5.82)
75.8 (3.94){
68.2 (3.55)
71.4 (6.25)
78.6 (3.91)
70.5 (3.61)
Anti-hypertensive medication, % (SE)
51.7 (5.41)
61.6 (4.59)
55.6 (4.35)
74.9 (5.38)
83.8 (3.77)
80.0 (3.08)
81.4 (5.34)
89.3 (3.25)
82.6 (3.06)
Wealth
Low
Medium
High
Low
Medium
High
Low
Medium
High
Systolic BP mean (SE)
133.7 (1.59) 135.9 (1.45) 136.4 (1.49)
133.8 (1.62)
131.9 (1.58)
128.6 (1.29){
131.6 (1.73) 130.7 (1.65) 128.2 (1.33)
Diastolic BP mean (SE)
81.1 (0.95)
81.0 (0.87)
82.2 (0.90)
85.6 (1.14)
85.5 (0.95)
83.5 (0.80)
83.9 (1.18)
84.6 (0.98)
83.2 (0.81)
BP ,140/90 mmHg, % (SE)
63.5 (4.61)
59.7 (4.61)
60.9 (4.37)
55.2 (4.74)
57.6 (4.39)
71.7 (3.95)`
60.7 (5.07)
60.4 (4.67)
72.5 (4.08){
BP ,150/90 mmHg, % (SE)
75.1 (4.34)
74.4 (4.04)
74.0 (3.96)
64.3 (4.84)
64.2 (4.36)
78.0 (3.51)`
69.3 (4.99)
67.6 (4.57)
78.9 (3.56)
Anti-hypertensive medication, % (SE)
57.1 (5.04)
59.3 (4.64)
53.8 (4.53)
71.2 (5.18)
80.5 (3.67)
86.0 (2.83)`
81.6 (4.70)
81.6 (3.86)
87.7 (2.73)
BP = Blood Pressure; SE = standard error; Values and significance testing control for age and sex, body mass index, vascular co-morbidity and smoking (age controlled at
57, female sex, non-smoker, no co-morbidity and BMI = 30.2 – the English mean in the age group. Main findings Significance tests compare the medium and high SEP groups with the
low within each country. {p,0.05. `p,0.01. doi 10 1371/journal pone 0083705 t004 Table 4. Blood pressure control and prescribing in hypertensive patients in US market-based v English NHS by socioeconomic-
position (aged 50-64 years). Table 4. Blood pressure control and prescribing in hypertensive patients in US market-based v English NHS by socioeconomic-
position (aged 50-64 years). BP = Blood Pressure; SE = standard error; Values and significance testing control for age and sex, body mass index, vascular co-morbidity and smoking (age controlled at
57, female sex, non-smoker, no co-morbidity and BMI = 30.2 – the English mean in the age group. Significance tests compare the medium and high SEP groups with the
low within each country. {p,0.05. `p,0.01. doi:10.1371/journal.pone.0083705.t004 pressure control. This may be especially relevant in a US sample
with no single insurance source, which is exposed to under-
insurance and co-payment [27]. Our data did not permit us to
determine which medications were prescribed (nor how many). Therefore, we could not assess the appropriateness of prescribing
(including combination therapy). burden disease and our study provides key insights related to its
management, it may not be appropriate to extrapolate findings to
other disease areas, particularly higher cost acute conditions. We
limited our study to white respondents in each country. This
allowed us to focus on national differences in health care insurance
and policies, without confounding from ethnically different
minority groups. This allows greater focus on the research
question, improves validity, but will limit the applicability of our
findings to the general population in both countries. Our data only
cover older adults, so findings may not be generalisable to all ages. Finally, the survey contained missing data for outcomes and
covariates. The nested survey design, however, combined with the
weights, ensure those with complete data remain representative. Strengths & Limitations Our study has potential limitations. We used patient-reported
diagnoses of hypertension. While this definition may underesti-
mate prevalence [5], we felt it more appropriate than one based on
blood pressure values, which will include undiagnosed cases. After
all, the health system is only able to manage disease when
diagnosed and recorded. Use of self-reporting may have influenced
our findings. Most notably, we may underestimate levels of
prescribing, although differences between countries are likely to be
minimal. We have data on only one aspect of hypertension
management; anti-hypertensive prescribing, however, this is a core
aspect of control, highlighted by clinical guidelines in both
countries. We did not have data on the class of antihypertensive
agent prescribed, or the number. These differences may contribute
to the cross-national differences in blood pressure control, and
may require further study. While hypertension is a common, high References Starfield B, Shi L, Macinko J (2005) Contribution of primary care to health
systems and health. Milbank Q 83: 457–502. 24. Majeed A, Banarsee R, Molokhia M (2009) Health Disparities and Community
Participation in England. The Journal of Ambulatory Care Management 32:
280. 8. Macinko J, Starfield B, Shi L (2007) Quantifying the health benefits of primary
care physician supply in the United States. International journal of health
services 37: 111–126. 25. McWilliams JM, Meara E, Zaslavsky AM, Ayanian JZ (2009) Differences in
control of cardiovascular disease and diabetes by race, ethnicity, and education:
U.S. trends from 1999 to 2006 and effects of medicare coverage. Ann Intern
Med 150: 505–515. 9. Caminal J, Starfield B, Sanchez E, Casanova C, Morales M (2004) The role of
primary care in preventing ambulatory care sensitive conditions. European
journal of public health 14: 246–251. 26. McWilliams JM, Meara E, Zaslavsky AM, Ayanian JZ (2007) Use of health
services by previously uninsured Medicare beneficiaries. N Engl J Med 357:
143–153. 10. Anderson GF, Hussey PS, Frogner BK, Waters HR (2005) Health spending in
the United States and the rest of the industrialized world. Health Aff (Millwood
24: 903–914. 27. Schoen C, Osborn R, How SK, Doty MM, Peugh J (2009) In chronic condition:
experiences of patients with complex health care needs, in eight countries, 2008. Health Aff (Millwood) 28: w1–16. 11. Gilmore I (2010) Prescription Charges Review Implementing Exemption from
Prescription Charges for People with Long Term Conditions London:
Department of Health, 2010, May 27. Report No.: DH 116366. 28. Link BG (2008) Epidemiological Sociology and the Social Shaping of Population
Health. J Health & Social Behaviour 49: 367–384. 12. Millett C, Everett CJ, Matheson EM, Bindman AB, Mainous AG, 3rd (2010)
Impact of Medicare Part D on seniors’ out-of-pocket expenditures on
medications. Arch Intern Med 170: 1325–1330. 29. Institute of Medicine (US) (2001) Committee on Quality of Health Care in
America. Crossing the quality chasm: A new health system for the 21st century. National Academies Press, 2001. 13. Patient Protection and Affordable Care Act of 2010. Public Law 111–148
section 6301 2010. 30. Lu JFR, Hsiao WC (2003) Does Universal Health Insurance Make Health Care
Unaffordable? Lessons From Taiwan. Health Affairs 22: 77–88. 14. Galobardes B, Shaw M, Lawlor DA, Lynch JW, Davey Smith G (2006)
Indicators of socioeconomic position (part 1). J Epidemiol Community Health
60: 7–12. 31. References 1. Heidenreich PA, Trogdon JG, Khavjou OA, Butler J, Dracup K, et al. (2011)
Forecasting the future of cardiovascular disease in the United States: a policy
statement from the American Heart Association. Circulation 123: 933–944. 17. Jones DW, Hall JE (2004) Seventh report of the Joint National Committee on
Prevention, Detection, Evaluation, and Treatment of High Blood Pressure and
evidence from new hypertension trials. Hypertension 43: 1–3. 2. Danaei G, Finucane MM, Lin JK, Singh GM, Paciorek CJ, et al. (2011)
National, regional, and global trends in systolic blood pressure since 1980:
systematic analysis of health examination surveys and epidemiological studies
with 786 country-years and 5.4 million participants. Lancet 377: 568–577. 18. Williams B, Poulter NR, Brown MJ, Davis M, McInnes GT, et al. (2004)
Guidelines for management of hypertension: report of the fourth working party
of the British Hypertension Society, 2004-BHS IV. J Hum Hypertens 18: 139–
185. with 786 country-years and 5.4 million participants. Lancet 377: 5 19. Millett C, Gray J, Bottle A, Majeed A (2008) Ethnic disparities in blood pressure
management in patients with hypertension after the introduction of pay for
performance. Ann Fam Med 6: 490–496. 3. Chobanian AV (2009) Shattuck Lecture. The hypertension paradox-more
uncontrolled disease despite improved therapy. N Engl J Med 361: 878–887. 4. Asch SM, Kerr EA, Keesey J, Adams JL, Setodji CM, et al. (2006) Who is at
greatest risk for receiving poor-quality health care? N Eng J of Med 354: 1147–
1156. 20. Roalfe AK, Holder RL, Wilson S (2008) Standardisation of rates using logistic
regression: a comparison with the direct method. BMC Health Serv Res 8: 275. 5. Banks J, Marmot M, Oldfield Z, Smith JP (2006) Disease and disadvantage in
the United States and in England. JAMA 295: 2037–2045. 21. Papke LE, Wooldridge JM (2005) A computational trick for delta-method
standard errors. Economics Letters 86: 413–417. 22. Yamey G (2010) Obama’s giant step towards universal health insurance. BMJ
340: c1674. 6. Avendano M, Glymour MM, Banks J, Mackenbach JP (2009) Health
Disadvantage in US Adults Aged 50 to 74 Years: A Comparison of the Health
of Rich and Poor Americans With That of Europeans. American Journal of
Public Health 99: 540–548. 23. Wilper AP, Woolhandler S, Lasser KE, McCormick D, Bor DH, (2009) Health
insurance and mortality in US adults. Am J Public Health 99: 2289–2295. 7. Conclusions Characteristics of both a single-payer health service and
universal health insurance coverage may facilitate high-quality,
equitable treatment. The expansion of Medicaid eligibility and
widening private insurance coverage under the ACA, indeed the
promotion of universal health coverage globally [37], may
encourage equitable management of chronic conditions such as
hypertension. Unlike disparities in disease prevalence which
appear influenced by, amongst other things, wider redistributive
social policy and the prevalence of disease risk factors [5,6], our
findings suggest disparities in hypertension control may be more
closely linked to health system structure, notably access to care
through insurance. Although our study demonstrates that not all
kinds of ‘‘universal’’ health insurance are alike, it supports the
ambition that expanding health insurance may reduce health
disparities. The English and American health care systems differ in many
ways, including the provision of universal coverage and overall
levels of expenditure. Our study demonstrates that hypertension
control in England matches that in the US in a sample without
universal health insurance, despite the evidence elsewhere of lower
overall expenditure. The two study samples covered by universal
health insurance had the lowest disparities in hypertension control,
and in one – in England – were entirely absent. The ACA’s vision
of improving access to healthcare may promote equitable care. Interestingly, while there were no indications of health care
disparities in the English system, some persisted in the US, even
when patients had insurance (either private insurance for patients
under 65 years or Medicare for older patients). The introduction
of universal coverage at the age of 65 years may not eliminate
disparities that become too entrenched; although may reduce
them. Again, the ACA’s expansion of Medicare and its individual
mandate – expanding private insurance coverage – may increase
equity. Alternately, other country-level differences, such as the
underlying structures or subtle challenges to care access, including
co-payment, may perpetuate disparities: If so, further system-level
changes may be required, which the ACA only begins to tackle. Author Contributions Conceived and designed the experiments: AD EV MH GN CM. Obtained
funding: AM CM. Administrative, technical, or material support: GN. Study supervision: CM. Statistical analysis: AD GN. Analysis and
interpretation of data: AD EV MH GN AM RW CM. Acquisition of
data: AD GN. Drafting of the manuscript: AD CM. Critical revision of the
manuscript for important intellectual content: AD EV MH GN AM RW
CM. Conceived and designed the experiments: AD EV MH GN CM. Obtained
funding: AM CM. Administrative, technical, or material support: GN. Study supervision: CM. Statistical analysis: AD GN. Analysis and
interpretation of data: AD EV MH GN AM RW CM. Acquisition of
data: AD GN. Drafting of the manuscript: AD CM. Critical revision of the
manuscript for important intellectual content: AD EV MH GN AM RW
CM. Health Insurance Coverage and Hypertension Control underlying causes of disease and poor control warrant greater
attention [36]. Impacts on policy and practice Hypertension control in both countries remains suboptimal;
blood pressure is controlled to clinical targets in just over a half of
patients aged 65 years and over. Given the association between
blood pressure control and good health outcomes [34], and despite
an array of efficacious medications [3], further efforts are required
to improve the management of hypertension in both countries. Chronic disease control may also be influenced by population
factors, including dietary behaviours and lifestyle [35]. Managing January 2014 | Volume 9 | Issue 1 | e83705 PLOS ONE | www.plosone.org 7 g
y J
yp
34. Lloyd-Jones DM, Evans JC, Levy D (2005) Hypertension in adults across the age
spectrum: current outcomes and control in the community. JAMA 294: 466–
472. 35. Levene LS, Baker R, Bankart MJ, Khunti K (2010) Association of features of
primary health care with coronary heart disease mortality. JAMA 304: 2028–
2034. cross-sectional findings from the British Women’s Heart and Health Study and
the British Regional Heart Study. J Hum Hypertens 20: 733–741. 36. Cappuccio FP, Capewell S, Lincoln P, McPherson K (2011) Policy options to
reduce population salt intake. BMJ 343: d4995. p p
J
37. Moreno-Serra R, Smith PC (2012) Does progress towards universal health
coverage improve population health? The Lancet 380: 917–923. Health Insurance Coverage and Hypertension Control References Wolf-Maier K, Cooper RS, Kramer H, Banegas JR, Giampaoli S, et al. (2004)
Hypertension treatment and control in five European countries, Canada, and
the United States. Hypertension 43: 10–17. 15. Marmot MG, Banks J, Blundell R, Lessof C, Nazroo J. Health, Wealth, and
Lifestyles of the Older Population in England — The 2002 English Longitudinal
Study of Ageing. London, England: Institute for Fiscal Studies; December 2003. 32. Mancia G, Bombelli M, Lanzarotti A, Grassi G, Cesana G, et al. (2002) Systolic
vs diastolic blood pressure control in the hypertensive patients of the PAMELA
population. Arch Intern Med 162: 582–586. 16. Juster FT, Suzman R (1995) An overview of the health and retirement study. Journal of Human Resources 30: S7–S56. 33. Patel R, Lawlor DA, Whincup P, Montaner D, Papacosta O, et al. (2006) The
detection, treatment and control of high blood pressure in older British adults: PLOS ONE | www.plosone.org January 2014 | Volume 9 | Issue 1 | e83705 January 2014 | Volume 9 | Issue 1 | e83705 8 Health Insurance Coverage and Hypertension Control Health Insurance Coverage and Hypertension Control Health Insurance Coverage and Hypertension Control January 2014 | Volume 9 | Issue 1 | e83705 PLOS ONE | www.plosone.org PLOS ONE | www.plosone.org 9
|
https://openalex.org/W4367859410
|
https://zenodo.org/records/7895523/files/DPTMS0501.pdf
|
English
| null |
SPIRITUALITY - HUMAN SPIRITUALITY AT THE STAGE OF HISTORICAL DEVELOPMENT
|
Zenodo (CERN European Organization for Nuclear Research)
| 2,023
|
cc-by
| 851
|
SPIRITUALITY - HUMAN SPIRITUALITY AT THE STAGE OF
HISTORICAL DEVELOPMENT
Usmanov Azizbek Zhuraevich
Andijan State Pedagogical Institute
methodology for teaching social and humanitarian Sciences
(fundamentals of spirituality) 2nd year master's degree:
https://doi.org/10.5281/zenodo.7895523
d h
h fl
f
f ll b
l d h When it is said that man is the flower of nature, of all being, it is implied that he
has this trait, that is, the possibility of being the owner of high spirituality. We
will not see this possibility in other creatures. While material objects give a
person physical food and energy, spirituality gives him spiritual food and
strength. Content only with material maintenance – inherent in unconscious and
soulless creatures. And the pursuit of spirituality is a virtue inherent only to the
human being, the owner of spirit and mind. Spirituality is such a complex social
phenomenon as the set of the mental and spiritual world of a person. Only those with high spirituality and enlightenment will live as glue, my country,
and it will be happiness for them to show kindness to others, to support those
who need help, and to guide them the right way. Those who have high
spirituality and compassion have hard work on their heads, make it easy for a
person who has fallen into grief, become a partner in his grief, become an
ointment for pain, make out a hojati, take care of orphans. In those who are the
owner of such a quality, national pride, a sense of national pride, orientality will
be strong. They consider it a glory for themselves to defend the honor of their
nation, the benefit of el-ulusi, to show their wealth, Soul, Sacrifice and examples
of heroism in the path of protection of the motherland if a security is born to
independence. It is also not very correct to conclude from the above that only
enlightened people would have a high spirituality. Looking at the history of
mankind, not all people with spirituality have always been enlightened. At the
same time we can see that not all people with knowledge, enlightenment were
highly spiritual. For such people, our people rightfully used the proverbial
phrase "become a scientist, but not a person", which expressed deep meaning in
itself. In other words, there are people who are not scientists. Without a
scientist, but with a high morale. There are scientists who do not deserve a
human breed. This one weighs. There is no benefit from them to the land. As our
people find and say, Rice will not be without kurmak. DEVELOPMENT OF PEDAGOGICAL TECHNOLOGIES IN
MODERN SCIENCES
International scientific-online conference DEVELOPMENT OF PEDAGOGICAL TECHNOLOGIES IN
MODERN SCIENCES
International scientific-online conference SPIRITUALITY - HUMAN SPIRITUALITY AT THE STAGE OF
HISTORICAL DEVELOPMENT
Usmanov Azizbek Zhuraevich
Andijan State Pedagogical Institute
methodology for teaching social and humanitarian Sciences
(fundamentals of spirituality) 2nd year master's degree:
https://doi.org/10.5281/zenodo.7895523
d h
h fl
f
f ll b
l d h SPIRITUALITY - HUMAN SPIRITUALITY AT THE STAGE OF
HISTORICAL DEVELOPMENT
Usmanov Azizbek Zhuraevich
Andijan State Pedagogical Institute
methodology for teaching social and humanitarian Sciences
(fundamentals of spirituality) 2nd year master's degree:
https://doi.org/10.5281/zenodo.7895523
d h
h fl
f
f ll b
l d h This phrase has been used
only in proportion to certain people of science whose spirituality is rare in life. Such people were viewed with contempt. On the contrary, those who took the
example of the people of Science, who harmoniously embodied spirituality and 5 DEVELOPMENT OF PEDAGOGICAL TECHNOLOGIES IN
MODERN SCIENCES
I
i
l
i
ifi
li
f International scientific-online conference International scientific-online conference enlightenment, dreamed that their children would become such people. Along
with the above phrase, our people also used the phrase "Be not a scientist." This
situation does not mean that spirituality and enlightenment deny each other. On
the contrary, spirituality and enlightenment are mutual, closely related factors
that persecute each other, promote the maturation of Man and society. As our
president said, We can see which period of our history we do not take, the
pursuit of ilmu enlightenment and high spirituality in our country has never
stopped, which has become evident even in the most difficult and complex times
as an immortal embodiment of the genius of our people. Our ancestors from
time immemorial considered the invaluable wealth of knowledge, education and
upbringing as the most basic condition and guarantee of human perfection and
prosperity of the nation. In conclusion, it can be said that any country has its own spirituality that
elevates this spirituality.So in our country,too,it is necessary to educate children
in the spirit of spirituality from a preschool institution of course it is also a
mistake to limit yourself to this, because this spirituality of ours goes back
several years, which means that it is also advisable to draw attention to families. In order for us to strengthen the Enlightenment, it is necessary to obtain more
knowledge, people who read books are definitely enlightened. References:
1. Milliy istiqlol g'oyasi: asosiy tushuncha va tamoyillar. T., «O'zbekiston»,
2000. References:
1. Milliy istiqlol g'oyasi: asosiy tushuncha va tamoyillar. T., «O'zbekiston»,
2000. 2. Falsafa qomusiy lug’at
|
https://openalex.org/W2227064212
|
https://www.cambridge.org/core/services/aop-cambridge-core/content/view/E2BB82618FFD5F1C843E36986CFA643F/S0955603600001112a.pdf/div-class-title-span-class-italic-maudsley-discussion-paper-11-should-mental-health-nurses-prescribe-span-by-k-gournay-r-gray-london-institute-of-psychiatry-21-pp-2001-4-00-pb-isbn-0-9500289-4-div.pdf
|
English
| null |
Maudsley Discussion Paper 11. Should Mental Health Nurses Prescribe? By K. Gournay, R. Gray, London: Institute of Psychiatry. 21 pp. 2001. £4.00 (pb). ISBN: 0-9500289-4
|
Psychiatric bulletin of the Royal College of Psychiatrists/Psychiatric bulletin
| 2,002
|
cc-by
| 1,449
|
Maudsley Discussion Paper
11. Should Mental Health
Nurses Prescribe? literature online, I was intrigued by the
notion of a book that compiles the
evidence-base for dementia. Searching
online for evidence from my desk at work,
the answer has always been ‘too much’ to
get hold of and at the same time ‘too
little’ that is relevant to my own practice. The central concept of this book is that
the author, working within the College
Research Unit, searches for evidence (of
vastly differing qualities) and lays it out in
a way that enables scrutiny. The book sets
out its search methodology clearly, cites
its sources and then presents ‘the
evidence’ with references and an attempt
to weigh its importance. The book’s remit
is to compile secondary research
evidence, not primary studies and papers. Therefore, it is a rather sad reflection on
the paucity of the evidence, that so much
of it is the national guidelines of the
College or the American Psychiatric Asso-
ciation, or evidence-based guidelines
relying on studies of moderate quality or
poorer, rather than systematic reviews
and critically appraised research summa-
ries. This is not the fault of the author,
who has done a creditable job to pull
these dry guidelines together. But the
larger point is that the College and its
publishing arm, Gaskell, have missed an
opportunity with this type of static
publication. The same material together
with the hugely useful website links and
critical appraisal resources should be
accessible on the College website or on
CD-ROM, even at a cost. Searching would
be easier, links to other resources would
be enhanced and the search could be as
contemporary as the day you access the
site (not as old as September 1998). The
shame is that, for all the author’s hard
work, the book is now way out of date. The idea of an evidence-based briefing is
a good one because of the approach, but
not in this format. Does the book help
reduce the personal effort required to get
on top of the evidence? Not really. Stephen Burton
Consultant and Senior Lecturer in
Old Age Psychiatry, Ladywell Unit, University Hospital By K. Gournay and R. Gray. London: Institute of Psychiatry. 21pp. 2001. »4.00 (pb). ISBN: 0-9500289-4 By K. Gournay and R. Gray. London: Institute of Psychiatry. 21pp. 2001. »4.00 (pb). ISBN: 0-9500289-4 By K. Gournay and R. Gray. London: Institute of Psychiatry. 21pp. 2001. »4.00 (pb). Columns Reviews Therefore, it is a rather sad reflection on
the paucity of the evidence, that so much
of it is the national guidelines of the
College or the American Psychiatric Asso-
ciation, or evidence-based guidelines
relying on studies of moderate quality or
poorer, rather than systematic reviews
and critically appraised research summa-
ries. This is not the fault of the author,
who has done a creditable job to pull
these dry guidelines together. But the
larger point is that the College and its
publishing arm, Gaskell, have missed an
opportunity with this type of static
publication. The same material together
with the hugely useful website links and
critical appraisal resources should be
accessible on the College website or on
CD-ROM, even at a cost. Searching would
be easier, links to other resources would
be enhanced and the search could be as
contemporary as the day you access the
site (not as old as September 1998). The
shame is that, for all the author’s hard
work, the book is now way out of date. The idea of an evidence-based briefing is
a good one because of the approach, but
not in this format. Does the book help
reduce the personal effort required to get
on top of the evidence? Not really. Columns Reviews Columns Reviews there is one minor quibble, it is that the
focus is heavily on psychiatric trainees. Safety concerns all health professionals, of
course, and the fundamental principles
espoused in this pack could apply to many
others in the NHS. One wonders if the
College has the energy to re-work the
material for a wider audience. If so, it
could be a real money-spinner. citing the ‘anti-medical’ model adopted by
some mental health nurses, thereby
assuming they would be reluctant to
prescribe. As for the medical profession,
nursing has a code of practice that is
subject to professional and legal
accountability. Therefore, it is slightly
presumptuous to consider that nurses
would fail in their duty of care in
prescribing to any greater degree than
psychiatrists. The same principles apply to
the rather convoluted discussions around
which nurses should prescribe which
drugs. This minor point aside, Safety in
Psychiatry reflects great credit on the
College and those involved in its produc-
tion. For anyone carrying responsibility for
the safety of trainees, there can be only
one message: buy, buy, buy! The paper only considers the role of
community mental health nurses, ignoring
the fact that this group comprises only
12% of qualified mental health nurses. The
more urgent need for nurses to be able to
initiate, titrate and alter medication often
lies in the hospital or group home envir-
onment. It would be useful to consider
these issues in relation to independent
and/or supplementary prescribing, and
the need for good collaborative working
and robust shared-care arrangements. David Newby
Consultant Psychiatrist, Leeds
Community & Mental HealthTeachingTrust literature online, I was intrigued by the
notion of a book that compiles the
evidence-base for dementia. Searching
online for evidence from my desk at work,
the answer has always been ‘too much’ to
get hold of and at the same time ‘too
little’ that is relevant to my own practice. The central concept of this book is that
the author, working within the College
Research Unit, searches for evidence (of
vastly differing qualities) and lays it out in
a way that enables scrutiny. The book sets
out its search methodology clearly, cites
its sources and then presents ‘the
evidence’ with references and an attempt
to weigh its importance. The book’s remit
is to compile secondary research
evidence, not primary studies and papers. Maudsley Discussion Paper
11. Should Mental Health
Nurses Prescribe? ISBN: 0-9500289-4 The discussion around evaluation of the
effectiveness of nurse prescribing suggests
using randomised controlled trials. Although this method is often considered
the gold standard of research, it may not
evaluate adequately the finer points of
prescribing practice or user satisfaction. Given the evidence base quoted in the
paper on current prescribing efficacy by
psychiatrists, it might be timely to consider
robust evaluation of all prescribing for
mental health patients, using both quanti-
tative and qualitative methods. In conclusion, much of this paper has
been superseded by recent events and
this is its main weakness. The Government
is moving fast on nurse prescribing. If this
paper is to have any real impact on the
current debate, it needs to link more
directly with the mainstream to avoid
being dragged along on the coat-tails of
directives from the Department of Health. This paper explores the context of mental
health nurse prescribing and covers such
considerations as rationale, supervision,
training and evaluation. Although well
researched and systematically argued, its
perspective is medically orientated and
this narrows the scope of the discussion. F.Winstanley
Senior Lecturer in Community
Nursing and Nurse Prescribing,
R. Dibblee
Community Mental Health Nurse, Suffolk College,
School of Health and Applied Sciences,The Ipswich
Hospital NHS Trust, Heath Road, Ipswich IP4 5PD The rationale is rightly argued from the
basis of patient need; but it is from a
public health and resource viewpoint,
rather than the more compelling one of
continuity of care. As in other areas of
chronic disease management, the
continuing contact of the nurse with the
patient informs the process of prescrip-
tion review and adjustment. Nurse
involvement could alter the focus from
treating patients pharmaceutically, to
putting medication in the context of a
care plan, which balances symptom
control against side-effects. EBB Evidence-Base Briefing:
Dementia. A Compilation
of Secondary Research
Evidence, Guidelines and
Consensus Statements
By Claire Palmer. London: Gaskell. 1999. 96 pp. »15.00 (pb). ISBN: 1-901242-35-8 EBB Evidence-Base Briefing:
Dementia. A Compilation
of Secondary Research
Evidence, Guidelines and
Consensus Statements
By Claire Palmer. London: Gaskell.
1999. 96 pp. »15.00 (pb).
ISBN: 1-901242-35-8 EBB Evidence-Base Briefing:
Dementia. A Compilation
of Secondary Research
Evidence, Guidelines and
Consensus Statements
By Claire Palmer. London: Gaskell. 1999. 96 pp. »15.00 (pb). ISBN: 1-901242-35-8 Pursuing best practice in our brave new
world of evidence-based medicine
demands considerable personal effort to
find out ‘what is known’. As an old age
psychiatrist, and regular searcher of the Stephen Burton
Consultant and Senior Lecturer in
Old Age Psychiatry, Ladywell Unit, University Hospital
Lewisham SE13 5QY A somewhat sweeping assumption is
made against nurse prescribing through 440 https://doi.org/10.1192/pb.26.11.440 Published online by Cambridge University Press
|
https://openalex.org/W4385852226
|
http://e-journal.janabadra.ac.id/index.php/JMIH/article/download/3047/pdf_117
|
Indonesian
| null |
PERLINDUNGAN HAK-HAK ANAK DALAM PERKARA PERMOHONAN DISPENSASI KAWIN SEBAGAI UPAYA PENCEGAHAN PERKAWINAN DIBAWAH UMUR
|
Kajian Hasil Penelitian Hukum
| 2,023
|
cc-by-sa
| 8,338
|
A R T I C L E I N F O Dalam penulisan tesis ini metode yang digunakan
adalah normatif empiris, yaitu penelitian terhadap
penerapan hukum normatif. Mengkaji implementasi
hukum normatif di Pengadilan Agama Purwokerto
terhadap peraturan perundang-undangan yang berkaitan
dengan perkara permonan dispensasi kawin, untuk
memastikan apakah penerapan hukum sudah sesuai
dengan
Undang-Undang
yang
berlaku
atau
ada
pertimbangan lain untuk memenuhi kebutuhan hukum
masyarakat. Article history:
Received (bulan) 10 Juli 2023
Accepted (bulan) 11 Juli, 2023
Available online 17 Juli, 2023
Kata Kunci:
Hak Anak, Dispensasi Kawin,
Perkawinan Bawah Umur. Keywords:
Children's Rights, Marriage
Dispensation, Underage Marriage. This is an open access article under the
CC BY-SA license. Copyright © 2022 by Author. Published
by
Magister
Hukum
Janabadra
Yogyakarta
https://doi.org/ Article history:
Received (bulan) 10 Juli 2023
Accepted (bulan) 11 Juli, 2023
Available online 17 Juli, 2023
Kata Kunci:
Hak Anak, Dispensasi Kawin,
Perkawinan Bawah Umur. Keywords:
Children's Rights, Marriage
Dispensation, Underage Marriage. This is an open access article under the
CC BY-SA license. Copyright © 2022 by Author. Published
by
Magister
Hukum
Janabadra
Yogyakarta
https://doi.org/ Article history:
Received (bulan) 10 Juli 2023
Accepted (bulan) 11 Juli, 2023
Available online 17 Juli, 2023 Article history:
Received (bulan) 10 Juli 2023
Accepted (bulan) 11 Juli, 2023
Available online 17 Juli, 2023 Kata Kunci:
Hak Anak, Dispensasi Kawin,
Perkawinan Bawah Umur. Keywords:
Children's Rights, Marriage
Dispensation, Underage Marriage. Dalam pembahasan tesis ini dijelaskan mengenai
prosedur dan faktor penyebab diajukannya perkara
dispensasi kawin yaitu faktor ekonomi, faktor pendidikan
yang rendah dan faktor hamil diluar nikah , pertimbangan
hakim dalam memutus perkara tersebut dikaitkan dengan
klausula alasan mendesak sebagaimana termuat dalam
pasal 7 ayat (2) Undang-Undang Nomor 16 Tahun 2019
tentang perubahan atas Undang-Undang Nomor 1 Tahun
1974 tentang perkawinan, proses persidangan sebelum
dan sesudah lahirnya Perma Nomor 5 Tahun 2019 tentang
Pedoman Mengadili Perkara Dispensasai Kawin serta
kemungkinan-kemungkinan
akibat
yang
akan
ditimbulkan terkait perlindungan hak-hak anak yang
mungkin dilanggar apabila perkara dispanssasi kawin
dikabulkan. Copyright © 2022 by Author. Published
by
Magister
Hukum
Janabadra
Yogyakarta https://doi.org/ Kata Kunci: Hak Anak, Dispensasi Kawin, Perkawinan
Bawah Umur. *Corresponding author.
E-mail addresses: antonisaid12@gmail.com 1 K. Wantjik Saleh, Hukum Perkawinan Indonesia, Ghalia indonesia, Jakarta, 2000, hlm.14.
2 CST. Kansil, Pengertian Ilmu Hukum dan Tata Hukum Indonesia, Balai Pustaka, Jakarta, 2018, hlm.230. A B S T R A C T This tesis used normative empirical method, study
about positif law, study of implementation normative law
in Purwokerto religious court about the constitution
related with matrimonial dispensation to ensure the
constitution has corresponding with positive law or judge
has enother consideration to fulfil public law. Jurnal Kajian Hasil Penelitian Hukum Vol. 7, No. 1, Tahun 2023 In this thesis explained abut procedure and causal factor
of matrimonial dispensation, economic factor, insufficient
education and unwed pregnancy judges consideration to
solve the case, urgent reason in chapter 7 verse (2)
constitution number 16 of 2019 abaut the chance of
constitution number 1 of 1974 abaut merriage, court
process before and after the rules of supreme court
number 5 of 2019 about court process of matrimonial
dispensation born and the possibility effect with the
children`s right protecting violated if matrimonial
dispensation case decided. Keywords: Children's Rights, Marriage Dispensation,
Underage Marriage. Keywords: Children's Rights, Marriage Dispensation,
Underage Marriage. Keywords: Children's Rights, Marriage Dispensation,
Underage Marriage. Available Online http://e-journal.janabadra.ac.id/index.php/JMIH/issue/archive 1 K. Wantjik Saleh, Hukum Perkawinan Indonesia, Ghalia indonesia, Jakarta, 2000, hlm.14.
2 Pendahuluan Undang-Undang Nomor 1 Tahun 1974 yang telah dirubah dengan Undang-
Undang Nomor 16 Tahun 2019 tentang perkawinan dinyatakan pada pasal 1 ayat (1)
”Perkawinan adalah ikatan lahir batin antara seorang pria dengan seorang wanita
sebagai suami isteri dengan tujuan membentuk keluarga (rumah tangga) yang bahagia
dan kekal berdasarkan Ketuhanan Yang Maha Esa”. Dalam melaksanakan perkawinan antara seorang laki-laki dengan seorang
perempuan ada batasan umur yang harus dipenuhi bagi calon pengantin. Pada pasal
7 ayat (1) Undang-Undang Nomor 1 Tahun 1974 tentang perkawinan disebutkan
bahwa batasan usia untuk laki-laki adalah 19 tahun dan untuk perempuan 16 tahun. Dengan “ikatan lahir-batin” dimaksudkan bahwa perkawinan itu tidak hanya cukup
dengan adanya “ikatan lahir” atau “ikatan batin” saja tapi harus keduanya. Suatu
“ikatan lahir” adalah ikatan yang dapat dilihat. Mengungkapkan adanya suatu
hubungan hukum antara seorang pria dan wanita untuk hidup bersama, sebagai
suami isteri, dengan kata lain dapat disebut “hubungan formil”.1) ”Ketentuan ini diadakan adalah untuk menjaga kesehatan suami isteri dan
keturunan, dan karena itu dipandang perlu diterangkan batas umur untuk
perkawinan dalam Undang-undang Perkawinan”.2) Lahirnya Undang-Undang Nomor 16 Tahun 2019 tentang perubahan atas
Undang-Undang Nomor 1 Tahun 1974 tentang perkawinan, pada pasal 7 ayat (1)
membawa perubahan yang signifikan pada batas usia perkawinan. Batasan usia
pernikahan bagi laki-laki dan perempuan disamakan menjadi 19 tahun. Disinilah
masalah mulai muncul karena perubahan batasan usia perkawinan tersebut. Masyarakat kita khususnya di daerah pedesaan ada sebagian yang masih mempunyai
padangan bahwa anak perempuan tidak perlu sekolah tinggi cukup sampai tingkat
SLTP saja sudah cukup untuk kemudian mereka akan dinikahkan kalau sudah ada Available Online http://e-journal.janabadra.ac.id/index.php/JMIH/issue/archive jodohnya. Sedangkan maksud dari perubahan Undang-Undang Perkawinan ini
diantaranya adalah agar anak bangsa dapat mengenyam pendidikan 12 tahun, atau
setidak-tidaknya sampai tingkat SLTA dan untuk mencegah perkawinan usia dini. Pendahuluan "Salah satu asas atau prinsip perkawinan yang ditentukan dalam Undang-undang
Perkawinan adalah bahwa calon suami isteri itu harus telah masak jiwa raganya
untuk dapat melangsungkan perkawinan agar dapat mewujudkan tujuan
perkawinan secara baik tanpa berakhir pada perceraian dan mendapat keturunan
yang baik dan sehat untuk itu harus dicegah adanya perkawinan yang masih di
bawah umur".3) "Salah satu asas atau prinsip perkawinan yang ditentukan dalam Undang-undang
Perkawinan adalah bahwa calon suami isteri itu harus telah masak jiwa raganya
untuk dapat melangsungkan perkawinan agar dapat mewujudkan tujuan
perkawinan secara baik tanpa berakhir pada perceraian dan mendapat keturunan
yang baik dan sehat untuk itu harus dicegah adanya perkawinan yang masih di
bawah umur".3) Pola pikir yang sudah mendarah daging ini mempengaruhi minat untuk
menuntut ilmu bagi anak perempuan bahwa mereka tidak perlu sekolah tinggi,
sehingga mempengaruhi perilaku mereka. Pada usia 15 tahun mereka sudah mulai
pacaran dengan harapan ketika lulus SLTP sudah mempunyai calon suami untuk
segera menikah dan hal itu tidak dilarang atau malah didukung oleh orang tua
mereka. Perubahan batas usia perkawinan yang diatur dalam pasal 7 (1) Undang-Undang
Nomor 16 Tahun 2019 menjadi 19 tahun bagi calon pengantin laki-laki maupun
perempuan diharapkan akan sempurna kematangan jiwa dan raganya dalam
memasuki bahtera kehidupan rumah tangga agar dapat tercipta tujuan perkawinan
dan dapat melahirkan keturunan yang sehat, selain itu juga dapat menjadi kendali
dalam menurunkan angka kematian ibu dan anak. Masing-masing suami dan isteri
dapat menjalankan hak dan kewajibannya masing-masing dengan sempurna sehingga
dapat mendorong sempurnanya tumbuh kembang anak secara maksimal. Kemajuan teknologi juga telah berpengaruh dalam mengubah keterbukaan
informasi publik secara global telah menciptakan dunia dalam genggaman, media
sosial telah menyentuh seluruh lapisan masyarakat sampai ke pelosok negeri. Hal
tersebut ikut andil dalam mempercepat pendewasaan anak-anak, karena adanya
globalisasi informasi yang tidak tersaring secara baik. Segala macam informasi dari
luar dengan mudah masuk ke seluruh penjuru negeri sehingga dengan cepat
mempengaruhi gaya hidup anak-anak sekarang namun tidak diimbangi dengan
kematangan pola pikir. Faktor sosiologis juga mempengaruhi pergaulan bebas pada
anak-anak, selain itu faktor intern keluarga, kurangnya pengawasan dan kasih sayang
serta perhatian dari orang tua akan menyebabkan anak terjerumus dalam sebuah
budaya pergaulan yang terlalu bebas. Akibatnya perzinaan sering terjadi dan
berujung pada kehamilan bagi anak-anak perempuan sebelum menikah. 3 Sudarsono, Hukum Perkawinan Nasional, cet.3, Rineka Cipta, Jakarta, 2005, hlm.7. Pendahuluan Pernikahan dini juga menimbulkan implikasi mengenai dilanggarnya hak-hak anak,
diantaranya hak untuk bermain, hak untuk mengenyam pendidikan dan diskriminasi
dari orang tua, sebagaimana tercantum dalam Undang-Undang perlindungan anak,
maka apabila dengan sengaja orang tua menikahkan anaknya dengan tujuan tertentu
dengan maksud untuk mengeksploitasi anak sehingga anak diabaikan hak-haknya,
maka telah melanggar pasal 26 ayat (1) huruf (c) Undang-undang Nomor 23 Tahun
2002 yang disempurnakan menjadi Undang-Undang Nomor 35 Tahun 2014 tentang
perlindungan anak, yang menyatakan bahwa orang tua berkewajiban dan
bertanggung jawab untuk mencegah terjadinya perkawinan pada usia anak-anak. Kematangan fisik dan psikologis bukan merupakan jaminan perkawinan akan
berjalan dengan mulus dan sempurnya, kematangan sosial dan ekonomi memegang
peranan penting dalam menjaga utuhnya sebuah rumah tangga. Seorang yang sudah
berani membentuk rumah tangga harus berani memberi nafkah kepada anak dan isteri
dan juga kematangan ekonomi belum tercapai akan muncul masalah pokok yaitu
kebutuhan hidup yang harus dipenuhi oleh seorang suami kepada keluarganya. Pernikahan dini juga menimbulkan implikasi mengenai dilanggarnya hak-hak anak,
diantaranya hak untuk bermain, hak untuk mengenyam pendidikan dan diskriminasi
dari orang tua, sebagaimana tercantum dalam Undang-Undang perlindungan anak,
maka apabila dengan sengaja orang tua menikahkan anaknya dengan tujuan tertentu
dengan maksud untuk mengeksploitasi anak sehingga anak diabaikan hak-haknya,
maka telah melanggar pasal 26 ayat (1) huruf (c) Undang-undang Nomor 23 Tahun
2002 yang disempurnakan menjadi Undang-Undang Nomor 35 Tahun 2014 tentang
perlindungan anak, yang menyatakan bahwa orang tua berkewajiban dan
bertanggung jawab untuk mencegah terjadinya perkawinan pada usia anak-anak. Pendahuluan Ketika hal ini sudah terjadi maka mau tidak mau suka tidak suka orang tua ikut
bertanggung jawab untuk menyelesaikan masalah ini, dengan cara yang sederhana
yaitu menikahkan anak di usia yang belum cukup, karena untuk menutupi aib
keluarga di mata masyarakat. Jurnal Kajian Hasil Penelitian Hukum Vol. 7, No. 1, Tahun 2023 Lahirnya Peraturan Mahkamah Agung Nomor 5 tahun 2019 tentang pedoman
mengadili perkara permohonan Dispensasi kawin mempunyai tujuan untuk
memberikan pedoman kepada para hakim secara lebih rinci dalam memahami
permohonan Dispensasi kawin agar tujuan dari Undang-Undang Nomor 16 Tahun
2019 tentang perubahan atas Undang-Undang Nomor 1 Tahun 1974 tentang
perkawinan dapat diwujudkan. Alasan yang mendesak menjadi salah satu syarat
untuk dapat dikabulkannya permohonan Dispensasi Kawin dan didukung dengan
bukti-bukti yang cukup adalah keadaan dimana tidak ada pilihan lain dan sangat
terpaksa untuk dilangsungkan perkawinan. Disamping meningkatnya perkara Dispensasi Kawin di Pengadilan Agama
Purwokerto, ada Permasalahan yang tidak kalah penting, yaitu bagaimana hakim
untuk menyelesaikan perkara Dispensasi Kawin, Teori hukum mana yang akan
dipakai oleh hakim dalam menyelesaikan perkara Dispensasi Kawin, Apakah
penerapan hukum positif atau Sociological Jurisprudence. Disatu sisi Hakim harus
memutus berdasarkan aturan yang sudah ada karena Indonesia menganut sistem Civil
Law. Hal ini juga bertujuan untuk mencapai maksud dari Undang-Undang Nomor 16
tahun 2019 tentang perubahan atas Undang-Undang Nomor 1 Tahun 1974 tentang
perkawinan yaitu untuk mencegah perkawinan usia dini, melindungi hak-hak anak
dan sebagainya. Akan tetapi disisi yang lain sebagian masyarakat kita belum siap
dengan perubahan ini. Didalam persidangan orang tua masih tetap ingin menikahkan
anaknya yang belum berusia 19 tahun dan celakanya si anak juga sudah berkeinginan
untuk menikah dan tidak ingin melanjutkan sekolahnya. Kematangan fisik dan psikologis bukan merupakan jaminan perkawinan akan
berjalan dengan mulus dan sempurnya, kematangan sosial dan ekonomi memegang
peranan penting dalam menjaga utuhnya sebuah rumah tangga. Seorang yang sudah
berani membentuk rumah tangga harus berani memberi nafkah kepada anak dan isteri
dan juga kematangan ekonomi belum tercapai akan muncul masalah pokok yaitu
kebutuhan hidup yang harus dipenuhi oleh seorang suami kepada keluarganya. 1. Dispensasi Kawin Sebuah Pengecualian Dari Aturan Umum Tinjauan Pustaka 1. Dispensasi Kawin Sebuah Pengecualian Dari Aturan Umum 1. Dispensasi Kawin Sebuah Pengecualian Dari Aturan Umum 1. Dispensasi Kawin Sebuah Pengecualian Dari Aturan Umum Available Online http://e-journal.janabadra.ac.id/index.php/JMIH/issue/archive Dispensasi kawin merupakan upaya bagi mereka yang ingin menikah
namun belum mencukupi batas usia untuk menikah yang telah ditetapkan oleh
pemerintah, sehingga bagi masyarakat yang beragama Islam orang tua dari anak
yang belum cukup umur tersebut bisa mengajukan dispensasi kawin ke
Pengadilan
Agama
melalui
proses
persidangan
terlebih
dahulu
agar
mendapatkan izin dispensasi perkawinan. Dispensasi kawin merupakan sebuah
kelonggaran hukum bagi mereka yang belum memenuhi syarat untuk
melangsungkan perkawinan secara hukum positif, oleh karena itu undang-
undang memberikan kewenangan kepada pengadilan untuk memberi dispensasi
kawin. Dalam peraturan perundang-undangan telah secara jelas disebutkan
mengenai batas usia perkawinan di Indonesia yaitu: Menurut Undang-Undang
Nomor 16 Tahun 2019 tentang perubahan atas Undang-Undang Nomor 1 Tahun
1974 tentang perkawinan, terdapat dalam Bab II syarat-syarat perkawinan pasal 6
ayat (2) yaitu : “Untuk melangsungkan perkawinan seorang yang belum mencapai
umur 21 (dua puluh satu) tahun harus mendapat izin kedua orang tua”. Selanjutnya dalam pasal 7 ayat (1) disebutkan “perkawinan hanya diizinkan jika
pria sudah mencapai umur 19 (sembilan belas tahun) dan pihak wanita sudah
mencapai umur 19 (sembilan belas) tahun. Selanjutnya pada ayat (2) disebutkan
bahwa “Dalam hal penyimpangan terhadap ayat (1) pasal ini dapat meminta
dispensasi nikah kepada Pengadilan Agama atau pejabat lain yang ditunjuk oleh
kedua orang tua pihak pria maupun pihak wanita. Dan pada ayat (3) “Ketentuan-
ketentuan mengenai keadaan salah seorang atau kedua orang tua tersebut dalam
pasal 6 ayat (3) dan (4) undang-undang ini, beraku juga dalam hal permintaan
dispensasi tersebut ayat (2) pasal ini dengan tidak mengurangi maksud pada pasal
6 ayat (2) tersebut. Batas usia nikah menurut Kompilasi Hukum Islam pada pasal 15 ayat (1):
“Untuk kemaslahatan keluarga dan rumah tangga perkawinan hanya bioleh
dilakukan calon mempelai yang telah mencapai umur yang ditetapkan dalam
pasal 7 Undang-Undang Nomor 1 Tahun 1974 yakni calon suami berumur
sekurang-kurangnya 19 (sembilan belas) tahun dan calon isteri sekurang-
kurangnya berumur 16 (enam belas) tahun. Dan pada ayatu (2): “Bagi calon
mempelai yang belum berumur 21 tahun harus mendapat izin yang sebagaimana
yang diatur dalam pasal 6 ayat (2), (3), (4) dan (5) Undang-Undang Nomor 1 Tahun
1974 tentang perkawinan. Tinjauan Pustaka Dalam Kitab Undang-Undang Hukum Perdata batasan usia menikah
disebutkan dalam bab IV tentang perkawinan pasal 29, yang berbunyi “Laki-laki
yang belum mencapai umur 18 (delapan belas) tahun penuh, tidak diperkenankan
mengadakan perkawinan. Namun jika ada alasan-alasan penting pemerintah
berkuasa menghapuskan larangan ini dengan memberikan dispensasi”. Jurnal Kajian Hasil Penelitian Hukum Vol. 7, No. 1, Tahun 2023 2. Penerapan hukum positif dalam menyelesaikan perkara permohonan Dispensasi
Kawin Sebagai Upaya Pencegahan Perkawinan Dibawah Umur dan Untuk
Melindungi Hak-Hak Anak. Aliran positivisme atau disebut juga dengan legisme hukum mulai
berkembang sejak abad pertegahan dan mempunyai pengaruh besar di berbagai
negara. Pengaruh positivisme juga sampai ke Indonesia, negara lain yang
dipengaruhi oleh positivisme hukum adalah Jerman dan diantara pakar hukum
Jerman yang mempertahankan positivisme ini adalah Paul Laband, Jellinek,
Rudolf van Jhering, Hans Nawiasky dan Hans Kelsen".4)
John Austin sebagai seorang tokoh pelopor hukum positif menyatakan
bahwa hukum adalah perintah dari penguasa negara dan hakikat dari hukum
adalah perintah itu. Menurut Austin hukum hukum dan perintah berjalan dari
atasan (superior) dan mengikat atau mewajibkan bawahan (inferior) Pihak
superior yang menentukan apa yang harus dilakukan atau diperbolehkan dan
kekuasaan superior memaksa orang lain untuk melakukannya. Hukum yang
sebenarnya
memuat
4
unsur
yaitu
perintah,
sanksi
kewajiban
dan
kedaulatan."Esensi dari madzhab positivisme adalah melihat hukum sebagai
sistem peraturan perundang-undangan yang dibuat dan diberlakukan oleh
negara secara formal (hukum positif). Walaupun sebuah nilai mempunyai
kekuatan mengikat dan dipatuhi oleh orang banyak seperti ajaran moral atau
ajaran agama tetapi tidak bisa dikatakan sebagai sebuah hukum kalau ia tidak
dirumuskan dalam peraturan yang dibuat oleh negara".5) Menurut John Austin,
"Hukum adalah perintah yang dibebankan untuk mengatur mahluk yang berfikir,
oleh karena itu yang membuat peraturan pembebanan tersebut harus mahlik yang
berfikir dari kalangan pemegang kekuasaan. Hukum juga harus dijalankan
dengan sistem yang logis meski bersifat tertutup. Selain itu secara tegas hukum
harus terpisah dari keadilan dan hukum tidak didasarkan pada nilai-nilai baik
atau buruk".6)
Berkaitan dengan pokok-pokok ajaran dalam positivisme hukum tersebut,
hakim dalam memeriksa perkara dispensasi kawin mengalami Permasalahan
dalam menetapkan teori hukum yang akan dipakai dalam memeriksa perkara ini 2. Penerapan hukum positif dalam menyelesaikan perkara permohonan Dispensasi
Kawin Sebagai Upaya Pencegahan Perkawinan Dibawah Umur dan Untuk
Melindungi Hak-Hak Anak. Aliran positivisme atau disebut juga dengan legisme hukum mulai
berkembang sejak abad pertegahan dan mempunyai pengaruh besar di berbagai
negara. 4 Zulkarnain, Hukum Kompetensi Peradilan Agama, cet I, Kencana Pranedamedia Grup, Jakarta, 2021, hlm.23.
5 Mukti Fajar ND dan Yulianto Achmad, op.cit., hlm.12.
6 Lili Rasjidi & Ira Thania, Dasar-Dasar Filsafat Dan Teori Hukum, PT Citra Aditiya Bakri, Bandung, 2007,
hlm.58. ,
p
s
d
g
,
,
G
p, J
, 0
,
3
5 Mukti Fajar ND dan Yulianto Achmad, op.cit., hlm.12. 6 Lili Rasjidi & Ira Thania, Dasar-Dasar Filsafat Dan Teori Hukum, PT Citra Aditiya Bakri, Bandung, 2007,
hlm.58. Zulkarnain, Hukum Kompetensi Peradilan Agama, cet I, Kencana Pranedamedia Grup, Jakarta, 2021, hlm Tinjauan Pustaka Pengaruh positivisme juga sampai ke Indonesia, negara lain yang
dipengaruhi oleh positivisme hukum adalah Jerman dan diantara pakar hukum
Jerman yang mempertahankan positivisme ini adalah Paul Laband, Jellinek,
Rudolf van Jhering, Hans Nawiasky dan Hans Kelsen".4) John Austin sebagai seorang tokoh pelopor hukum positif menyatakan
bahwa hukum adalah perintah dari penguasa negara dan hakikat dari hukum
adalah perintah itu. Menurut Austin hukum hukum dan perintah berjalan dari
atasan (superior) dan mengikat atau mewajibkan bawahan (inferior) Pihak
superior yang menentukan apa yang harus dilakukan atau diperbolehkan dan
kekuasaan superior memaksa orang lain untuk melakukannya. Hukum yang
sebenarnya
memuat
4
unsur
yaitu
perintah,
sanksi
kewajiban
dan
kedaulatan."Esensi dari madzhab positivisme adalah melihat hukum sebagai
sistem peraturan perundang-undangan yang dibuat dan diberlakukan oleh
negara secara formal (hukum positif). Walaupun sebuah nilai mempunyai
kekuatan mengikat dan dipatuhi oleh orang banyak seperti ajaran moral atau
ajaran agama tetapi tidak bisa dikatakan sebagai sebuah hukum kalau ia tidak
dirumuskan dalam peraturan yang dibuat oleh negara".5) Menurut John Austin,
"Hukum adalah perintah yang dibebankan untuk mengatur mahluk yang berfikir,
oleh karena itu yang membuat peraturan pembebanan tersebut harus mahlik yang
berfikir dari kalangan pemegang kekuasaan. Hukum juga harus dijalankan
dengan sistem yang logis meski bersifat tertutup. Selain itu secara tegas hukum
harus terpisah dari keadilan dan hukum tidak didasarkan pada nilai-nilai baik
atau buruk".6) Berkaitan dengan pokok-pokok ajaran dalam positivisme hukum tersebut,
hakim dalam memeriksa perkara dispensasi kawin mengalami Permasalahan
dalam menetapkan teori hukum yang akan dipakai dalam memeriksa perkara ini. Apakah akan menerapkan hukum positif dengan segala konsekuensinya demi
kepastian hukum atau menerapkan pendekatan hukum sosiologis untuk
mewujudkan asas kemanfaatan hukum dalam masyarakat. Drs. Asnawi, S.H., M.H. memperikan penjelasan bahwa dalam hal
menjatuhkan penetapan pada perkara dispensasi Kawin, hakim tunggal memiliki Available Online http://e-journal.janabadra.ac.id/index.php/JMIH/issue/archive (dua) pertimbangan-pertimbangan
yang
diklasifikasikan
menjadi
2
(dua)
pertimbangan, yaitu: 1) Pertimbangan Hukum 1) Pertimbangan Hukum Pertimbangan hukum di sini berarti ketika hakim menjatuhkan
penetapannya harus sesuai dengan dalil-dalil dan bukti-bukti hukum yang
diajukan oleh para pihak (Pemohon). Bukti-bukti yang biasa disyaratkan
menurut undang-undang adalah bukti surat-surat dan sakai.: Bukti saksi yang
biasa dihadirkan oleh Pemohon dalam persidangan adalah 2 (dua) orang atau
lebih. Sebagaimana dalam pertimbangannya, hakim juga berdasarkan hukum
Islam yakni menolak bahaya didahulukan atas mendatangkan kebaikan dan
kemadharatan harus dihilangkan yang pada dasarnya setiap insan tidak
diizinkan mengadakan suatu kemadharatan, baik berat maupun ringan
terhadap dirinya atau terhadap orang lain. Pada prinsipnya kemadharatan
harus dihilangkan, tetapi dalam menghilangkan kemadharatan itu tidak boleh
sampai menimbulkan kemadharatan lain baik ringan apalagi lebih berat. Namun, bila kemadharatan itu tidak menimbulkan kemadharatan yang lain
maka haruslah memilih kemadharatan yang relatif lebih ringan dari yang telah
terjadi. Madharat yang dimaksudkan dalam pertimbangan majelis hakim
adalah bilamana anak Pemohon tidak dinikahkan dikhawatirkan akan
menambah perbuatan dosa dan juga berpotensi terjadi pernikahan di bawah
tangan (nikah sirri) yang jutstru akan mengacaukan prosesproses hukum yang
akan terjadi berikutnya atau mengacaukan hak-hak hukum anak yang
dilahirkan. Berdasarkan Undang-undang Nomor 48 Tahun 2009 tentang Kekuasaan
Kehakiman, hakim dalam menetapkan sebuah permohonan harus sesuai
dengan nilai-nilai hukum dan rasa keadilan yang hidup di dalam masyarakat. Hal tersebut diatur dalam Pasal 5 ayat (1) yang menyatakan: “Hakim dan
hakim konstitusi wajib menggali, mengikuti, dan memahami nilai-nilai hukum
dan rasa keadilan yang hidup dalam masyarakat”. Bagi sebagian besar
masyarakat, pernikahan seringkali dianggap sebagai solusi alternatif dalam
menyelesaikan Permasalahan sosial yang terjadi di tengah-tengah masyarakat,
seperti menikahkan anak yang telah hamil di luar nikah. Hal tersebut
dilakukan untuk menutupi aib atau rasa malu pihak keluarga. Dari hasil
penelitian menunjukkan di Pengadilan Agama Purwokerto, hakim seringkali
mengabulkan permohonan dispensasi kawin karena hubungan di luar nikah
(marriage by accident), dengan pertimbangan perempuan yang hamil tanpa
suami akan dihina dan dikucilkan oleh masyarakat. Sehingga akan
mengakibatkan perempuan tersebut menghindar dan menjauh dari pergaulan
sosial. Hal tersebut juga berpotensi terjadi pada anak yang akan dilahirkan. Jurnal Kajian Hasil Penelitian Hukum Vol. 7, No. 7 Soerjono Soekanto, Pokok-Pokok Sosiologi Hukum , Rajawali Pers, Depok, 1988, hlm.43. 1) Pertimbangan Hukum 1, Tahun 2023 Menurut Roscoe Pound, tokoh dari aliran sociological jurisprudence
menjelaskan bahwa “Hukum harus dilihat atau dipandang sebagai suatu
lembaga kemasyarakatan yang berfungsi untuk memenuhi kebutuhan-
kebutuhan sosial dan tugas dari ilmu hukum untuk mengembangkan suatu
kerangka yang mena kebutuhan-kebutuhan sosial dapat terpenuhi secara
maksimal”.7) Keadaan sosial masyarakat dan adat untuk menikahkan anak di usia
muda serta asas kemanfaatan hukum merupakan salah satu pertimbangan
hakim dalam mengabukan permohonan dispensasi kawin, pertimbangan ini
mengacu kepada aliran sociological jurisprudence, dimana hukum yang
digunakan dalam tataran law in action harus dibedakan dengan hukum dalam
tataran law in book. Perbedaan ini dapat diterapkan dalam seluruh bidang
hukum. Pelaksanaan law in book ke dalam law in action harus memperhatikan
kepentingan masyarakat dan memenuhi kebutuhan hukum yang diperlukan
masyarakat. Hukum merupakan salah satu instrumen pengendalian sosial
(social control), hukum selalu menghadapi pertentangan dari kepentingan-
kepentingan. Dalam tataran law in action hakim Pengadilan Agama pada
belum dapat menerapkan hukum positif dalam permohonan dispensasi
kawin. Efek yang ditimbulkan akan besar jika hukum positif dipaksakan untuk
diterapkan. Apabila
hukum
positif
benar-benar
diterapkan
dalam
permohonan dispensasi kawin maka kemungkinan sebagian besar perkara
dispensasi kawin akan ditolak oleh hakim karena Undang-Undang sudah
mengatur mengenai batasan umur pernikahan dan dispensasi kawin adalah
menyimpangi dari aturan tersebut. Padahal sejak diundangkannya Undang-
Undang Nomor 16 Tahun 2019 tentang perubahan atas Undang-Undang
Nomor 1 Tahun 1974 tentang perkawinan, perkara permohonan dispensasi
kawin meningkat secara signifikan di seluruh Pengadilan Agama di Indonesia
dan untuk saat ini hakim Pengadilan Agama sebagian besar masih
menerapkan pendekatan hukum sosiologis. Available Online http://e-journal.janabadra.ac.id/index.php/JMIH/issue/archive Metode Penelitian ini adalah penelitian normatif yaitu penelitian dari penerapan
terhadap keberlakuan hukum normatif. Metode pendekatan yang digunakan
adalah metode pendekatan yuridis yang berbasis pada hukum normatif
Pendekatan yuridis normatif adalah memandang atau menelaah obyek yang diteliti
dari aspek hukum. Sumber data dalam penelitian ini berupa bahan-bahan hukum
yang terdiri dari bahan hukum primer, sekunder dan tersier. Bahan-bahan hukum
tersebut meliputi buku-buku dan artikel ilmiah mengenai hukum perdata dan
pidana, hukum perkawinan, perlindungan dan hak-hak anak, dan teori hukum. Data yang diperoleh dari bahan-bahan hukum diperkuat dengan data yang Available Online http://e-journal.janabadra.ac.id/index.php/JMIH/issue/archive diperoleh dari wawancara kepada narasumber. Narasumber adalah seseorang yang
memberikan pendapat atas obyek yang kita teliti. Narasumber dalam penelitian ini
adalah hakim Pengadilan Agama Purwokerto selaku praktisi dalam menyelesaikan
perkara permohonan Dispensasi Kawin. Data yang diperoleh dianalisis secara
kualitatif. 1. Proses Pemeriksaan Perkara Dispensasi Kawin. 1. Proses Pemeriksaan Perkara Dispensasi Kawin. Perkara dispensasi kawin merupakan jenis perkara perdata voluntair
(permohonan). Perkara voluntair mempunyai karakteristik yang menjadikannya
berbeda dengan perkara contentiosa (gugatan), dimana perkara permohonan
merupakan perkara yang hanya diajukan oleh pihak pemohon saja dan tidak ada
lawan. Prosedur penerimaan perkara Dispensasi Kawin di Pengadilan Agama
mengacu pada Pedoman Pelaksanaan Tugas dan Administrasi Peradilan Agama,
atau yang lazim disebut sebagai Buku II, dimana sistem pelayanan perkara di
pengadilan agama/mahkamah syar’iyah menggunakan sistem meja, yaitu sistem
kelompok kerja terdiri dari: Meja I (termasuk Kasir), Meja II, dan Meja III. Proses pendaftaran pekara Dispensasi Kawin dimulai dari pihak Pemohon
Dispensasi Kawin mengajukan permohonan kepada Ketua Pengadilan Agama
secara tertulis yang ditandatangani oleh Pemohon atau Kuasanya yang sah
bilamana Pemohon berhalangan (Pasal 6 ayat (5) Undang-Undang Nomor 1
Tahun 1974). Namun bagi Pemohon yang tidak dapat membaca dan menulis
dapat mengajukan permohonannya secara lisan di hadapan Ketua Pengadilan
Agama dalam prakteknya dilakukan dihadapan Hakim Pengadilan, permohonan
tersebut dicatat oleh hakim yang ditunjuk (Pasal 120 HIR/Pasal 144 RBg). Tahap
berikutnya permohonan didaftarkan dalam buku register dan diberi nomor
perkara setelah Pemohon membayar biaya panjar perkara yang besarannya sudah
ditentukan oleh Pengadilan Agama (Pasal 121 ayat (4) HIR/Pasal 145 ayat (4)
RBg). Dalama perndaftaran tersebut untuk kelancaran proses persidangan
selanjutnya
Pihak
Pemohon
disyaratkan
pula
membawa
kelengkapan
administrasi lainnya seperti Fotokopi Kartu Tanda Penduduk (KTP), Kartu
Keluarga, Akta Nikah, Akta Kelahiran Anak dan Ijazah Terakhir; Hakim setelah menerima berkas perkara kemudian mempelajari dan
memeriksa berkas perkara. Selanjutnya, membuat penetapan hari sidang (PHS)
yang isinya menetapkan hari dan tanggal serta jam pelaksaan sidang perkara serta
memerintahkan kepada Jurusita/Jurusita Pengganti untuk memanggil para pihak
agar datang menghadap pada hari, tanggal, dan jam yang telah ditentukan. Kepada para pihak diberitahukan kepadanya bahwa dapat mempersiapkan bukti-
bukti yang diajukan dalam persidangan. Namun dalam implementasinya bukti-
bukti surat sudah diserahkan kepada petugas ketika pendaftaran perkara. Setelah Jurnal Kajian Hasil Penelitian Hukum Vol. 7, No. 1, Tahun 2023 persidangan dibuka dan dinyatakan terbuka untuk umum oleh Ketua Majelis,
maka para pihak berperkara dipanggil ke ruang persidangan. Lalu Ketua Majelis
memeriksa identitas para Pemohon. Selanjutnya, Majelis Hakim berusaha
menasehati pemohon, anak pemohon dan calon anak pemohon dengan
memberikan penjelasan tentang akibat hukum bilaman pernikahan dilakukan
belum cukup umur dan agar menunda pernikahannya. Apabila penasehatan oleh
majelis hakim tidak berhasil, selanjutnya Hakim membacakan surat permohonan
pemohon. Available Online http://e-journal.janabadra.ac.id/index.php/JMIH/issue/archive 1. Proses Pemeriksaan Perkara Dispensasi Kawin. Hakim memulai pemeriksaan dengan pertanyaan-pertanyaan yang
diajukan kepada pemohon, anak pemohon dan calon anak pemohon secara
bergantian. Hakim melanjutkan pemeriksaan bukti surat (tertulis), dan pemohon
menyerahkan bukti tertulisnya. Setelah Majelis Hakim memeriksa bukti surat
(tertulis) yang diajukan oleh Pemohon. Selanjutnya, Majelis Hakim melanjutkan
dengan pemeriksaan saksi-saksi. Setelah Hakim memeriksa saksi-saksi yang dihadirkan oleh Pemohon di
persidangan dan dianggap pemeriksaan sudah cukup, selanjutnya Pemohon
dipersilahkan
untuk
mengajukan
kesimpulan,
dan
setelah
Pemohon
menyampaikan
kesimpulan
Hakim
menyatakan
sidang
diskors
untuk
musyawarah. Pemohon, anak pemohon dan calon anak pemohon diperintahkan
ke luar dari ruang persidangan. Setelah musyawarah selesai, skors dicabut dan
pemohon dipanggil kembali masuk ke ruang persidangan, kemudian dibacakan
penetapan, isi penetapan dapat berupa permohonan dikabulkan, ditolak atau
tidak dapat diterima sesuai dengan realita pemeriksaan persidangan. Keharusan dihadirkan di persidangan dimaksudkan untuk mendapatkan
penasihatan dari hakim pemeriksa perkara dispensasi kawin. Penasihatan yang
dilakukan oleh hakim pemeriksa perkara dispensasi kawin adalah dengan tujuan
untuk memberikan gambaran kepada pihak-pihak mengenari resiko dari
perkawinan di bawah umur, terutama yang berkaitan dengan 1) Kemungkinan berhentinya pendidikan anak 2) Keberlanjutan anak dalam menempuh wajib belajar 12 tahun; 3) Belum siapnya organ reproduksi anak; 4) Dampak sosial ekonomi dan psikologis bagi anak; 5) Potensi perselisihan dan kekerasan dalam rumah tangga. 6) Dispensasi Kawin Sebelum dan Sesudah lahirnya Perturan Mahkamah Agung
nomor 5 tahun 2019 6) Dispensasi Kawin Sebelum dan Sesudah lahirnya Perturan Mahkamah Agung
nomor 5 tahun 2019 Sebelum lahirnya Perturan Mahkamah Agung nomor 5 tahun 2019 tentang
pedoman mengadili perkara permohonan Dispensasi Kawin, Pengadilan belum
mempunyai kesamaan secara keseluruhan dalam penyelesaian permohonan
dispensasi kawin karena tidak ada aturan khusus yang mengatur secara jelas
mengenai apa saja ketentuan yang dapat dijadikan alasan agar dispensasi kawin Available Online http://e-journal.janabadra.ac.id/index.php/JMIH/issue/archive dikabulkan. Prosedur penyelesaian permohonan dispensasi kawin, tidak berbeda
dengan prosedur perkara permohonan lainnya. Prosedur tersebut tercantum
dalam Buku II tentang Pedoman Pelaksanaan Tugas dan Administrasi Peradilan
Agama, Calon suami istri yang belum mencapai usia 19 dan 16 tahun yang ingin
melangsungkan perkawinan, harus mengajukan permohonan dispensasi kawin
oleh orang tua calon suami/ istri di Pengadilan Agama atau Mahkamah syariah,
prosedur lainnya sama dengan permohonan di pengadilan lainnya. 1. Proses Pemeriksaan Perkara Dispensasi Kawin. Setelah lahirnya Peraturan Mahkamah Agung Nomor 5 Tahun 2019
keberadaan Perturan Mahkamah Agung tersebut hakim dalam memeriksa
perkara dispensasi kawin mendapatkan payung hukum yang kuat dalam
mengadili dan memutus perkara dispensasi kawin, Peraturan Mahkamah Agung
Nomor 5 Tahun 2019 tentang pedoman mengadili perkara permohonan
dispensasi kawin hadir yang pada intinya bertujuan untuk menjamin pelaksanaan
sistem peradilan yang melindungi hak anak, mengidentifikasi apakah ada atau
tidaknya paksaan yang melatarbelakangi permohonan dispenasasi perkawinan
anak, mewujudkan standarisasi proses mengadili dispensasi kawin, dan
meningkatkan tanggung jawab orang tua untuk mencegah perkawinan anak . 2. 2. Faktor-Faktor penyebab permohonan Dispensasi Kawin, perlindungan hak-hak
anak dan kondisi kejiwaan anak 2. Faktor-Faktor penyebab permohonan Dispensasi Kawin, perlindungan hak-hak
anak dan kondisi kejiwaan anak Dari hasil wawancara penulis dengan hakim Pengadilan Agama
Purwokerto yang bernama Dra.Teti Himati, diperoleh data bahwa ada 3 masalah
yang menjadi faktor penyebab diajukannya permohonan dispensasi kawin di
Pengadilan Agama Purwokerto yaitu: Kondisi ekonomi yang lemah mempengaruhi pola pikir orang tua untuk
menikahkan anak perempuannya di usia muda dengan harapan dapat
memperoleh suami yang cukup mapan sehingga beban orang tua menjadi lebih
ringan. Orang tua lebih memilih anaknya untuk dinikahkan dengan orang yang
mampu secara finansial dalam usia yang belum cukup lalu dimintakan
dispensasi ke Pengadilan Agama. Kesulitan ekonomi orang tua memicu banyak kasus dalam persoalan
kemiskinan yang menyebabkan perkawinan dibawah umur, sebagai imbasnya
anak tidak melanjutkan sekolah karena sudah menjadi ibu rumah tangga muda. Dalam kenyataannya tidak semua harapan tersebut dapat terwujud karena
setelah menikah ternyata suami ja tidak mempunyai kemampuan yang cukup
dalam hal ekonomi. Bagi masyarakat kalangan ekonomi lemah, menikahkan anak di usia
muda merupakan sebuah pilihan, bukan untuk kebahagiaan si anak namun
demi meringankan beban pribadi orang tua. Semakin cepat anak menikah maka
semakin ringan beban mereka. Perbuatan orang tua yang menikahkan anaknya Jurnal Kajian Hasil Penelitian Hukum Vol. 7, No. 1, Tahun 2023 karena untuk meringankan beban ekonomi keluarga telah melanggar pasal 11
Undang-undang Nomor 23 Tahun 2022 tentang Perlindungan anak, yaitu Hak
atas perlindungan dari perlakuan diskriminasi, eksploitasi baik ekonomi
maupun seksual, penerlantaran, kekejaman, kekerasan dan penganiayaan,
ketidakadilan dan perlakuan salah lainnya karena untuk meringankan beban ekonomi keluarga telah melanggar pasal 11
Undang-undang Nomor 23 Tahun 2022 tentang Perlindungan anak, yaitu Hak
atas perlindungan dari perlakuan diskriminasi, eksploitasi baik ekonomi
maupun seksual, penerlantaran, kekejaman, kekerasan dan penganiayaan,
ketidakadilan dan perlakuan salah lainnya 2) Faktor Pendidikan Yang Rendah )
g
Selanjutnya Dra. Available Online http://e-journal.janabadra.ac.id/index.php/JMIH/issue/archive 1. Proses Pemeriksaan Perkara Dispensasi Kawin. Teti Himati, hakim Pengadilan Agama Purwokerto
menjelaskan bahwa faktor rendahnya pendidikan juga mempengaruhi pola
pikir orang tua dan sudah menjadi mindset di daerah pedesaan bahwa urusan
perempuan adalah dapur, sumur dan kasur. Tidak ada gunanya perempuan
sekolah tinggi karena nantinya hanya akan menjadi seorang ibu yang melayani
suami dan mengurusi anak-anak di rumah. Tidak ada keinginan untuk
menyekolahkan anak ke jenjang yang lebih tinggi meskipun orang tua mampu
untuk membiayai. Pendidikan sangat mempunyai andil yang besar. Jika seorang anak
putus sekolah pada usia wajib sekolah kemudian mengisi waktu dengan
bekerja. Saat bekerja anak tersebut sudah merasa bisa mandiri untuk
menghidupi diri sendiri. Atas dasar tersebut seorang anak akan melakukan
pernikahan dibawah umur. Sedangkan orang tua yang juga berpendidikan
rendah belum memahami arti penting dari pendikan tentu akan mendukung
keinginan anak untuk menikah usia muda. Perbuatan orang tua yang menikahkan anaknya karena alasan
pendidikan yang rendah dan tidak mau menyekolahkan anaknya ke jenjang
yang lebih tinggi telah melanggar pasal 1 angka 2 Undang-Undang Nomor 23
Tahun 2002 tentang Perlindungan anak, dijelaskan bahwa Perlindungan anak
adalah segala kegiatan untuk menjamin dan melindungi anak dan hak-haknya
agar dapat hidup,timbuh,berkembang dan berpartisipasi secara optimal sesuai
dengan harkat dan martabat kemanusiaan serta mendapat perlindungan dari
kekerasan dan diskriminasi; 3) Faktor Hamil Di Luar Nikah. Faktor hamil di luar nikah juga merupakan salah satu penyebab
diajukannya permohonan dispensasi kawin di Pengadilan Agama. Alasan ini
termasuk kategori dalam alasan mendesak seperti disebutkan dalam pasal 7
ayat (2) Undang-Undang Nomor 16 Tahun 2019. Alasan Hamil di luar nikah
menjadi faktor yang cukup dominan di Pengadilan Agama. Adanya pergaulan
bebas yang terjadi saat ini mempengaruhi hakim dalam memutus perkara
dispensasi kawin, orang tua yang khawatir anaknya terjerumus dalam
perbuatan zina dan meminimalisir dampak yang akan timbul Dalam kasus ini
hakim biasanya lebih mempertimbangkan asas kemaslahatan dan kemanfaatan
serta untuk meminimalisir dampak yang akan timbul jika dispensasi kawin Available Online http://e-journal.janabadra.ac.id/index.php/JMIH/issue/archive tidak dikabulkan. Jangan sampai timbul persepsi masyarakat kalau
mengajukan permohonan dispensasi kawin harus hamil terlebih dahulu maka
akan dikabulkan Konvensi hak anak Pasal 13 sampai dengan Pasal 15 yang didalamnya
terkandung arti yang cukup luas antara lain, setiap anak berhak
mengemukakan pendapatnya dan mengekspresikan apa isi hati nuraninya dan
memiliki kebebasan untuk memilih dan menentukan apa pilihan agamanya. 1. Proses Pemeriksaan Perkara Dispensasi Kawin. Secara tidak langsung pasal ini telah melindungi hak asasi manusia yang
dimiliki oleh seorang anak, sehingga orang tua tidak dapat memaksakan
kehendaknya agar si anak ini merasa dikorbankan demi adat atau reputasi dari
orang tuanya. Orang tua yang mempunyai wawasan luas semestinya mau
mendengarkan keinginan dari anaknya , namun dalam kaitannya dengan
faktor penyebab perkara dispenasi kawin diantaranya adalah rendahnya
tingkat pendidikan orang tua, sehingga tidak semua orang tua memiliki pikiran
yang terbuka mungkin secara normalnya boleh-boleh saja orang tua
menyampaikan keinginannya kepada sang anak tetapi orang tua juga harus
menyadari bahwa sang anak juga memiliki cita-cita dan memiliki gambaran
untuk hidupnya dimasa depan. Namun demikian di persidangan hakim tidak
mudah untuk mendeteksi apakahperkara dispensasi kawin yang diajukan
adalah kehendak anak atau kehendak orangtuanya. Meskipun anak sudah
diminta keterangan di persidangan tanpa didapningi orang tuanya namun di
luar persidangan hakim tidak mengetahui apakah anak sudah di doktrin untuk
menjawab pertanyaan hakim atau bahkan anak diancam atau paling tidak
ditekan oleh orang tuanya agar menyatakan bahwa perkawinan ini adalah
keinginan dirinya bukan paksaan orang tuanya. Dampak paksaan untuk melakukan pernikahan dini pada anak ada
banyak hal, contohnya nya depresi, gangguan psikologi, minder, dan lain
sebagainya. Hal ini dapat menyebabkan depresi bagi sang anak yang belum
menganggap dirinya cakap sebagai orang dewasa. Beban berat bagi para hakim
untuk memutus perkara dispensasi kawin. Selain harus memperhatikan hak-
hak anak yang harus dilindungi juga harus memperhatikan segi kejiwaan anak
ketika anak melangsungkan perkawinan dibawah umur.Perlindungan
terhadap kejiwaan anak juga diatur oleh konvensi hak anak, selain
memperjuangkan hak-hak yang diinginkan oleh anak atau pendapat pendapat
yang ingin dikemukakan oleh anak, negara juga melindungi keamanan dan dan
kesehatan dari anak tersebut sehingga megingat anak adalah harapan masa
depan bangsa, harus dilindungi dari segala aspek agar dapat tercipta bibit
unggul sumber daya manusia untuk menjadi calon penerus masa depan bangsa
dan Negara. Penerapan hukum positif dalam perkara dispensasi kawin 3. Penerapan hukum positif dalam perkara dispensasi kawin Jurnal Kajian Hasil Penelitian Hukum Vol. 7, No. 1, Tahun 2023 Penerapan hukum positif dalam perkara dispensasi kawin Jika dikaitkan
dengan foktor penyebab diajukannya perkara dispensasi kawin diatas penulis
dalam melakukan penelitian ke Pengadilan Agama Purwokerto mendapat
informasi dari Hakim Pengadilan Agama Purwokerto yang bernama Drs. 8 Abdul Manan, Aneka Masalah Hukum Perdata Islam di Indonesia, Kencana, Jakarta, 2008, hlm.11. Available Online http://e-journal.janabadra.ac.id/index.php/JMIH/issue/archive 1. Proses Pemeriksaan Perkara Dispensasi Kawin. Asnawi, SH., MH, yang menerangkan bahwa pelaksanaan hukum positif
terhadap perkara dispensasi kawin di Pengadilan Agama Purwokerto sulit
untuk diterapkan karena adanya frase "alasan sangat mendesak disertai bukti-
bukti pendukung yang cukup” dalam pasal 7 ayat (2) Undang-Undang Nomor
16 Tahun 2019 tentang perubahan atas Undang-Undang Nomor 1 Tahun 1974
tentang perkawinan merupakan celah untuk mengabulkan perkara dispensasi
kawin. Padahal menurut Abdul Manan Pembatasan usia perkawinan bagi anak
bertujuan untuk menekan semaksimal mungkin agas dapat mencegah
perkawinan anak.8) Alasan sangat mendesak dengan disertai bukti-bukti yang cukup
menurut Drs. Asnawi, S.H., M.H. menjelaskan bahwa hal itu merupakan
pembaharuan hukum yang termuat dalam Undang-Undang Nomor 16 Tahun
2019 yang tidak terdapat dalam Undang-Undang Nomor 1 Tahun 1974. Dengan
tujuan untuk menekan angka pernikahan di bawah umur. Namun karena
masyarakat kita belum siap dengan pembaharuan tersebut maka terjadilan
kenaikan permohonan dispensasi Kawin di Pengadilan Agama Purwokerto. Keadaan sosial masyarakat dan adat untuk menikahkan anak di usia muda
serta asas kemanfaatan hukum merupakan salahsatu pertimbangan hakim
dalam mengadili dispensasi kawin di persidangan dengan presepsi masing-
masing. Dari keadaan yang telah diuraikan di atas, maka dapat di pahami bahwa
pelaksanaan hukum positif dalam perkara dispensasi kawin masih menemui
kendala dari sisi masyarakat Indonesia sendiri. Hakim dalam hal ini memahami
bahwa penerapan hukum positif belum bisa dipaksakan. Sebagian besar hakim
di Pengadilan Agama dalam memeriksa dispensasi kawin menggunakan
pendekatan hukum sosiologis, dimana faktor kultur dan budaya masyarakat
kita lebih bisa menerima. Data yang diperoleh di Pengadilan Agama Purwokerto jumlah perkara
Dispensasi Kawin pada tahun 2017 sebanyak 71 perkara dan pada tahun 2018
sebanyak 76 pekara. Setelah berlaku Undang-Undang Nomor 16 Tahun 2019
tentang perubahan atas Undang-Undang Nomor 1 Tahun 1974 tentang
perkawinan dimana terdapat perubahan batasan umur menikah meniadi 19
tahun baik untuk pria maupun wanita maka jumlah perkara permohonan
dispensasi kawin meningkat tajam. Perkara Dispensasi Kawin tahun 2020 Available Online http://e-journal.janabadra.ac.id/index.php/JMIH/issue/archive sebanyak 301 perkara dan tahun 2021 sebanyak 294 perkara. Ini berarti
peningkatan jumlah perkara mencapai lebih dari 400%. Hakim dalam memutuskan suatu permohonan dispensasi kawin tidak
jarang menemui masalah yang dilematis. Di satu sisi hakim merupakan
lembaga yudikatif yang harus menegakkan keadilan berdasarkan peraturan
perundang-undangan yang berlaku, namun di sisi lain hakim memandang
bahwa pada kenyataannya sebagian besar orang tua atau wali yang
mengajukan permohonan dispensasi karena sesuatu hal yang menurut norma
sosial dan hukum yang berlaku di masyarakat anak tersebut harus dinikahkan. 1. Proses Pemeriksaan Perkara Dispensasi Kawin. Apalagi ketika anak tersebut sudah hamil, ketika menemui keadaan seperti itu
maka hakim tentu memilih untuk mengabulkan permohonan dispensasi yang
diajukan sebab Hakim memandang bahwa anak yang di dalam kandungan
tersebut harus memiliki perlindungan hukum dan memiliki status yang jelas,
jelas nasabnya, jelas siapa orang tuanya. Jadi kedepannya anak yang di dalam
kandungan tersebut ketika telah lahir ke dunia bukan hanya memiliki
hubungan hukum dengan ibunya saja melainkan memiliki hubungan hukum
juga dengan ayahnya. 4. Kebebasan Hakim Dalam Memutus Perkara Dispensasi Kawin
Meningkatnya perkara dispensasi kawin di Penghadilan Agama telah
mengalami peningkatan yang signifikan. Salah satu penyebab dari peningkatan
tersebut adalah dari faktor hukumnya itu sendiri. Hukum yang berlaku di
Indonesia memberikan peluang besar terjadinya perkawinan anak. Di satu sisi
Indonesia telah memberlakukan Undang-Undang Nomor 23 Tahun 2002
tentang perlindungan anak yang mendefinisikan anak adalah seseorang yang
berlum berusia 18 (delapan belas) tahun, namun disasi yang lain Undang-
Undang Nomor 16 Tahun 2019 tentang perubahan atas Undang-undang
Nommor 1 Tahun 1974 tentang perkawinan memberi celah kepada anak untuk
melakukan perkawinan. Dari penelitian yang penulis lakukan di Pengadilan Agama Purwokerto
pada tahun 2021 maka penulis mendapatkan data mengenai perkara
permohonan dispensasi kawin selama tahun 2017, 2018, 2020 dan 2021, jumlah
perkara permohonan dispensasi kawin sebanyak 742 perkara. Dari perkara
yang masuk tersebut dikabulkan sebanyak 712 perkara, dicabut sebanyak 20
perkara, gugur sebanyak 8 perkara dan ditolak sebanyak 1 perkara. Data
tersebut menunjukkan bahwa tingkat dikabulkannya perkara dispensasi kawin
di Pengadilan Agama Purwokerto sangat tinggi yaitu 95,9%, sedangkan sisanya
sebanyak 4,1% dicabut, gugur dan ditolak. Penerapan hukum positif di Pengadilan Agama Purwokerto tergambar
dari perkara yang dicabut, gugur dan ditolak yaitu sebesar 4,1%. Keberanian
hakim untuk memeriksa dispensasi kawin secara positivis masih sangat minim, Jurnal Kajian Hasil Penelitian Hukum Vol. 7, No. 1, Tahun 2023 padahal bila ini diterapkan merupakan bentuk dari pencegahan permasalahan
yang dikhawatirkan akan lebih besar dampaknya. Tidak begitu signifikan
membicarakan masalah yang urgen dan mendesak untuk dilaksanakan
pernikahan dibawah umur karena menjadi kontra produktif terhadap
masyarakat yang tinggi tingkat ketaatan hukumnya namun rendah kesadaran
hukumnya. “Dalam perspektif sosiologi hukum kesadaran hukum berbeda
dengan ketaatan hukum. Kesadaran hukum sangat tinggi levelnya karena
bersumber dari hati nurani, dalam konteks dispensasi kawin kelompok yang
sadar hukum akan menginterpretasikan bahwa lebih baik tidak melakukan
perkawinan anak. 9 Mardi Candra,op.cit., hlm.127.
10 Romli Atmasasmita, Teori Hukum Integtratif, CV Mandar Maju, Bandung, 2019, hlm.34-35. 9 Mardi Candra,op.cit., hlm.127. 10 Romli Atmasasmita, Teori Hukum Integtratif, CV Mandar Maju, Bandung, 2019, hlm.34-35. 9 Mardi Candra,op.cit., hlm.127.
10 Romli Atmasasmita, Teori Hukum Integtratif, CV Mandar Maju, Bandung, 2019, hlm.34-35. 1. Proses Pemeriksaan Perkara Dispensasi Kawin. Adapun kelompok ketaatan hukum menginterpretasikan
dengan peluang yang perlu dimanfaatkan dan menjadikannya sebagai payung
hukum untuk melakukan pekawinan anak”.9) Merujuk pada pendapat Roscou Pound yang menyatakan bahwa “Law
as social enginering” dapat dipahami bahwa itu adalah konsep hukum masa
depan memuliki visi yang jelas tentang bagaimana menyelesaikan berbagai
konflik kepentingan dalam kehidupan masyarakat dan hubungan negara
dengan individu. Pernyataan Pound ini ditafsirkan Mochtar Kusuma Atmadja
dalam konteks pembangunan hukum nasional di Indonesia yang intinya
bagaimana memerankan hukum sebagai sarana pembaruan masyarakat.10) Sistem hukum di Indonesia yang menganut civil law, memberikan
kebebasan pada hakim untuk mengadili perkara sesuai dengan hati nuraninya
masing-masing, hal ini yang membuat para hakim kesulitan menerapkan
positivisme hukun pada dispensasi kawin karena adanya klausul “alasan
sangat mendesak”. Padahal klausul "alasan sangat mendesak” tersebut sangat
sulit dibuktikan kecuali calon mempelai perempuan sudah dalam keadaan
hamil. Pernikahan dibawah umur juga mempunyai resiko yang sangat tinggi
khususnya bagi anak perempuan. Dengan adanya perkawinan maka akan
muncul hak dan kewajiban dari masing-masing suami dan isteri sehingga
kemungkinan besar sebuah pernikahan dini juga akan diikuti dengan
kehamilan. Resiko pereraian pada pernikahan dibawah umur juga tinggi
karena belum matangnya pola pikir dan tingkat emosi yang tinggi, dan secara
sosial anak perempuan cenderung akan mengalami kesulitan yang lebih jika
terjadi perceraian. Ketika terjadi perceraian soerang anak perempuan akan
kehilangan haknya sebagai anak, dia sudah dianggap dewasa, tidak lagi
menjadi tanggung jawab orang tuanya dan akan menghalanginya untuk
melanjutkan pendidikan formal. Mencari pekerjaan adalah salah satu solusinya Available Online http://e-journal.janabadra.ac.id/index.php/JMIH/issue/archive namun karena tingkat pendidikan formal yang rendah juga akan menyulitkan
anak perempuan untuk mendapatkan pekerjaan yang layak. 5. Hak-Hak Anak Yang Terlindungi Dengan Penerapan Hukum Positif Dalam
Dispensasi Kawin. Harus diakui ketika hukum positif diterapkan dalam perkara dispensasi
kawin paling tidak akan dapat mengurangi terjadinya perkawinan dibawah
umur di Indonesia dan ada banyak hak-hak anak yang terlindungi, seperti
terlindungi hak tumbuh kembangnya,baik secara fisik maupun psikis,
terlindungi
masa
depannya
dengan
tidak
dibebani
dengan
ikatan
perkawinan,terlindungi
hak
untuk
menentukan
masa
depannya
sendiri,terlindungi dari hak untuk mengambil keputusan dan secara medis
terlindungi dari ancaman penyakit yang rentan menyerang perkawinan
dibawah umur seperti kanker serviks, kegagalan kehamilan yang dapat
mengakibatkan kematian bagi ibu. Ketika sudah terjadi perkawinan, maka merupakan suatu kewajiban bagi
masing-masing pasangan untuk memberikan nafkah batin. Dalam keadaan
isteri masih dibawah umur maka ada kemungkinan terjadi pemaksaan
hubungan suami isteri dimana isteri belum siap untuk melakukan itu. 11 Nurul Qomar, Hak Asasi Manusia Dalam Negara Hukum Demokrasi, Sinar Gafika, Jakarta, 2013, hlm.141.
12 Muladi, Hak Asasi Manusia-Hakekat, Konsep Dan Implikasinya Dalam Perspektif Hukum Dan Masyarakat,
Rafika Aditama, Bandung, 2009, hlm.160. 1. Proses Pemeriksaan Perkara Dispensasi Kawin. Akibatnya akan muncul tindak kekerasan dalam rumah tangga (KDRT) yang
berkaitan dengan hubungan seksual dan ini sangat merugikan pihak
perempuan yang melakukan pernikahan dibawah umur. Dalam teori hukum pidana dikenal adanya pertanggungjawaban pidana
dengan menganut asas kesalahan, artinya ketika sudah terpenuhi untus-unsur
pidana dan kesalahan maka seseorang dapat dijatuhi hukuman pidana. Hal ini
menjadi dilema ketika seorang suami ingin melakukan hubungan suami isteri
dengan isterinya yang sah namun karena ketidaksiapan isteri yang masih
dibawah umur sehingga timbul pemaksaan hubungan yang seharusnya bukan
merupakan sebuah tindak pidana namun dalam keadaan isteri yang belum siap
organ reproduksinya akan berubah menjadi sebuah tindak pidana dengan
terjadinya pemaksaan hubungan seksual. Dasar dari pertanggungjawaban pidana adalah adanya kesalahan,
sedangkan kesalahan ada dua macam yaitu sengaja (dolus) dan kelalaian (culpa). Yang menjadi pertanyaan adalah apakah seorang suami yang ingin
berhubungan dengan isterinya merupakan sebuah kesalahan atau kelalaian
atau tidak kedua-duanya. Hal tersebut bisa dihindari ketika pesangan suami
isteri sudah dalam keadaan masak jiwa dan raganya dan sudah dapat
memenuhi hak dan kewajibannya masing-masing, tertu saja salah satu syarat
untuk dapat terpenuhi hal tersebut adalah umur yang cukup. Perbuatan pidana dalam keluarga yang mungkin terjadi juga tidak mudah
dibuktikan bila dikaitkan dengan syarat pemidanaan yaitu Mens Rea dan Actus Jurnal Kajian Hasil Penelitian Hukum Vol. 7, No. 1, Tahun 2023 Reus. Adalah hal yang janggal dalam keadaan normal, bila dalam sebuah
keluarga seorang suami mempunyai niat untuk melakukan tindak pidana
kekerasan kepada isteri namun yang ada adalah tindakan suami yang ingin
berhubungan dengan isterinya dimana isteri dalam keadaan belum siap baik
fisik maupun psikis. Niat dari suami adalah bukan untuk menyakiti isterinya
namun untuk memberikan nafkah batin dan memenuhi kebutuhan biologis baik
suami maupun isteri, sedangkan isteri belum siap untuk itu, akibatnya isteri
merasakan penderitaan atau luka akibat dari perbuatan suami tersebut. Dalam
hal ini unsur mens rea tidak terpenuhi karena tidak ada niat suami untuk
menyakiti isterinya maskipun ada kemungkinan unsur actus reus terpenuhi,
sehingga kembali isteri yang menjadi korban. Oleh karena itulah lembaga Dispensasi Kawin berupaya untuk membantu
meminimalisir kemungkinan terjadinya tindak pidana dalam keluarga dan
untuk melindungi hak-hak perempuan. Selain itu jika dilihat dari sudut
pandang hukum positif di Indonesia (hukum nasional) setidak-tidaknya ada
beberapa aturan yang dilanggar dengan pelaksanaan pernikahan dibawah
umur, diataranya adalah: 1) Undang-Undang Nomor 16 Tahun 2019 tentang perubahan atas Undang-
Undang Nomor 1 Tahun 1974 tentang perkawinan. 1. Proses Pemeriksaan Perkara Dispensasi Kawin. 1) Undang-Undang Nomor 16 Tahun 2019 tentang perubahan atas Undang-
Undang Nomor 1 Tahun 1974 tentang perkawinan. 2) Ki
b U d
U d
H k
Pid
(
l 288 KUHP) 2) Kitab Undang-Undang Hukum Pidana, (pasal 288 KUHP); 3) Undang-Undang Nomor 23 Tahun 2002 tentang perlindungan anak. 4) Undang-Undang Nomor 12 Tahun 2022 tentang tindak pidana kekerasan
seksual. Namun dalam keadaan perempuan yang sudah hamil, harus diperhatikan
pula hak-hak anak yang masih dalam kandungan. Sehubungan dengan janin
yang merupakan anugerah yang diberikan Tuhan Yang Maha Esa dalam
Undang-undang No. 39 Tahun 1999 Tentang Hak Asasi Manusia yang juga
memuat terkait perlindungan hak janin.11) Dalam pasal 53 dinyatakan bahwa
“Setiap anak sejak dalam kandungan, berhak untuk hidup, mempertahankan
hidup, dan meningkatkan taraf kehidupannya”. Dalam hukum memang tidak dijelaskan secara detail mengenai hak janin,
lebih menjelaskan tentang anak, akan tetapi janin merupakan cikal bakal anak
yang nantinya menjadi subyek hukum atau pelaku hukum. Menurut batasan
usia, untuk hukum tertulis yang terdapat didalam hukum perdata berbeda-
beda
tergantung
dari
perundang-undangannya.12)
Sehingga
ketika
permohonan dispensasi kawin keadaan calon mempelai perempuan sudah
dalam keadaan hamil, maka harus dipertimbangkan hak-hak anak yang ada
dalam kandungan tersebut untuk mengabulkan atau menolak permohonan Available Online http://e-journal.janabadra.ac.id/index.php/JMIH/issue/archive dispensasi kawin. Hakim juga harus mempertimbangkan bahwa anak
khususnya anak perempuan merupakan manusia yang paling lemah, pada
umumnya anak sangat tergantung pada orang dewasa dalam segala aspek
kehidupannya, anak sangat rentan terhadap tindak kekerasan yang dilakukan
orang dewasa dan secara psikoligis masih labil. Memberikan perlindungan
kepada anak bukan semata-mata tanggungjawab orang tua namun sebagai
seorang hakim juga harus bertanggungjawab untuk melindungi anak sesuai
dengan bidangnya dan sesuai dengan kebutuhan anak pada saat itu. Daftar Pustaka Abdul Manan. (2008). Aneka Masalah Hukum Perdata Islam di Indonesia. CST. Kansil. (1989). Pengertian Ilmu Hukum dan Tata Hukum Indonesia. Jakarta: Balai
Pustaka. CST. Kansil. (1989). Pengertian Ilmu Hukum dan Tata Hukum Indonesia. Jakarta: Balai
Pustaka. Hadi Sutrisno. (1993). Metodologi Resarch. Jilid I cet. Ke 24. Yogyakarta: Andi Ofset. K. Wantjik Saleh. (2000). Hukum Perkawinan Indonesia. Jakarta: Ghalia indonesia. Mardi Candra. (2021). Pembaharuan Hukum Dispensasi Kawin Dalam Sistem Hukum
Indonesia. Jakarta: Kencana Prenadamedia Group. Muladi. (2009). Hak Asasi Manusia-Hakekat, Konsep Dan Implikasinya Dalam Perspektif
Hukum Dan Masyarakat. Bandung: Rafika. Nurul Qomar. (2013). Hak Asasi Manusia Dalam Negara Hukum Demokrasi. Jakarta: Sinar
Gafika. Peter Mahmud Marzuki. (2005). Penelitian Hukum. Cet.6. Jakarta: Kencana Prenada
Media Grup. omli Atmasasmita. (2019). Teori Hukum Integtratif. Bandung: CV Mandar Maju. Soerjono Soekanto. (1988). Pokok-Pokok Sosiologi Hukum. Jakarta: Rajawali Pers. Sudarsono. (2005). Hukum Perkawinan Nasional. Cetakan Ketiga. Jakarta: Rineka Cipta. Zulkarnain. (2021). Hukum Kompetensi Peradilan Agama. Cetakan Pertama. Jakarta:
Kencana Pranedamedia Group. Lili Rasjidi & Ira Thania. (2007). Dasar-Dasar Filsafat Dan Teori Hukum. Bandung: PT
Citra Aditiya Bakri. Mukti Fajar ND & Yulianto Achmad. (2019). Dualisme Penelitian Hukum Normatif dan
Empiris. Yogyakarta: Pustaka Pelajar. Kesimpulan Masalah yang dihadapi hakim Pengadilan Agama Purwokerto dalam menyelesaikan
perkara permohonan Dispensasi Kawin adalah akan menerapkan hukum positif atau
menerapkan menggunakan pendekata hukum sosiologis. Antara memenuhi
kebutuhan hukum masyarakat atau turut mendukung semangat dari diterbitkannya
Undang-Undang Nomor 16 Tahun 2019 tentang perubahan atas Undang-Undang
Nomor 1 Tahun 1974 tentang perkawinan yang diapresiasi dengan terbitnya Perturan
Mahkamah Agung Nomor 5 Tahun 2019 tentang pedoman mengadili perkara
permohonan dispensasi kawin yang bertujuan untuk mengurangi pernikahan
dibawah umur dan melindungi hak-hak anak, karena masyarakat Indonesia sebagian
masih dalam tataran taat hukum belum sadar hukum. Disamping itu sejak terbitnya
Peraturan Mahkamah Agung nomor 5 Tahun 2019 tentang pedoman mengadili
permohonan dispensasi kawin telah terjadi perubahan norma yang cukup signifikan
dalam mengadili permohonan dispensasi kawin. Konsep tentang kepentingan terbaik
bagi anak (best interets of child) secara bertahap menjadi paradigma utama dalam
dispensasi kawin. Kalau Dispensasi Kawin diputus dengan hukum positif maka
akan banyak perkara yang ditolak oleh hakim karena Undang-Undang melarang
perkawinan dibawah umur 19 tahun, namun hal ini bertentangan dengan rasa
keadilan masyarakat yang menginginkan kepastian hukum. Sedangkan bila diputus
dengan mempertimbangkan sosiologi hukum, maka banyak perkara yang dikabulkan
dan ini menyalahi semangat Undang-Undang Nomor 16 Tahun 2019 tentang
perubahan atas Undang-Undang Nomor 1 Tahun 1974 tentang perkawinan adalah
untuk mengurangi pernikahan dibawah umur dan melindungi hak-hak anak
Sebagian besar hakim di Pengadian Agama dalam memeriksa permohonan dispensasi
kawin masih menerapkan pendekatan hukum sosiologis karena pertimbangan-
pertimbangan tertentu yang berkaitan dengan adat, budaya, kebutuhan masyarakat
dan keadaan calon pengantin yang mengharuskan hakim untuk mengabulkan
permohonan dispensasi kawin, Diperlukan keberanian para hakim untuk
menyelesaikan perkara permohonan dispensasi kawin dengan menerapkan hukum
positif karena bila hukum positif benar-benar diterapkan dalam perkara permohonan
dispensasi kawin dapat menekan laju perkawinan dibawah umur dan penerapan
hukum positif dapat membantu melindungi hak-hak anak dan akan mencegah Jurnal Kajian Hasil Penelitian Hukum Vol. 7, No. 1, Tahun 2023 kemungkinan terjadinya tindak pidana pada anak andaikata terjadi pernikahan
dibawah umur. Juga untuk memberikan pembelajaran kepada masyarakat bahwa
tidak semua permohonan Dispensasi Kawin yang diajukan akan dikabulkan oleh
Pengadilan Agama. Peraturan Perundang-Undangan Undang-Undang Dasar Negara Republik Indonesia Tahun 1945. Undang-Undang Dasar Negara Republik Indonesia Tahun 1945. Kitab Undang-Undang Hukum Pidana
Kitab Undang-Undang Hukum Perdata
Undang-Undang Nomor 1 Tahun 1974 tentang Perkawinan. Available Online http://e-journal.janabadra.ac.id/index.php/JMIH/issue/archive
Undang-Undang Dasar Negara Republik Indonesia Tahun 1945. Kitab Undang-Undang Hukum Pidana
Kitab Undang-Undang Hukum Perdata
Undang-Undang Nomor 1 Tahun 1974 tentang Perkawinan. Undang-Undang Nomor 39 Tahun 1999 tentang Hak Asasi manusia Undang-Undang Dasar Negara Republik Indonesia Tahun 1945. Kitab Undang-Undang Hukum Pidana
Kitab Undang-Undang Hukum Perdata
Undang-Undang Nomor 1 Tahun 1974 tentang Perkawinan. Undang-Undang Nomor 39 Tahun 1999 tentang Hak Asasi manusia Kitab Undang-Undang Hukum Pidana Kitab Undang-Undang Hukum Perdata Undang-Undang Nomor 39 Tahun 1999 tentang Hak Asasi manusia Available Online http://e-journal.janabadra.ac.id/index.php/JMIH/issue/archive Undang-Undang Nomor 48 Tahun 2009 tentang Kekuaaan Kehakiman. Undang-Undang Nomor 48 Tahun 2009 tentang Kekuaaan Kehakiman. Undang-Undang Nomor 11 Tahun 2012 tentang Sistem Peradilan Pidana Anak; Undang-Undang Nomor 11 Tahun 2012 tentang Sistem Peradilan Pidana Anak; Undang-Undang Nomor 35 Tahun 2014 tentang perubahan atas Undang-Undang
Nomor 23 Tahun 2002 tentang Perlindungan Anak. Undang-Undang Nomor 35 Tahun 2014 tentang perubahan atas Undang-Undang
Nomor 23 Tahun 2002 tentang Perlindungan Anak. Undang-Undang Nomor 16 Tahun 2019 tentang perubahan atas Undang-Undang
Nomor 1 Tahun 1974 tentang Perkawinan. Undang-Undang Nomor 16 Tahun 2019 tentang perubahan atas Undang-Undang
Nomor 1 Tahun 1974 tentang Perkawinan. Undang-Undang Nomor 16 Tahun 2019 tentang perubahan atas Undang-Undang
Nomor 1 Tahun 1974 tentang Perkawinan. Peraturan Mahkamah Agung Nomor 5 Tahun 2019 tentang Pedoman Mengadili
Perkara Permohonan Dispensasi Kawin. Peraturan Mahkamah Agung Nomor 5 Tahun 2019 tentang Pedoman Mengadili
Perkara Permohonan Dispensasi Kawin. Peraturan Mahkamah Agung Nomor 5 Tahun 2019 tentang Pedoman Mengadili
Perkara Permohonan Dispensasi Kawin.
|
https://openalex.org/W3181109755
|
https://www.sciengine.com/doi/pdf/8EEB1241C9224F3A8FEEF962BA3F5E9C
|
Chinese
| null |
Determination of hydrazine in prednisolone by derivatization-gas chromatography-triple quadrupole mass spectrometry
|
Sepu/Chinese journal of chromatography
| 2,021
|
cc-by
| 815
|
∗收稿日期：２０２１⁃０３⁃０１
∗通讯联系人．Ｔｅｌ：（０１０）５８７１７２７９，Ｅ⁃ｍａｉｌ：ｇｕａｎｇｈｕｉ＿９＠１６３．ｃｏｍ．
基金项目：北京市改革与发展专项（２０２１ＺＬ０１２２）．
Ｆｏｕｎｄａｔｉｏｎｉｔｅｍ：ＢｅｉｊｉｎｇＭｕｎｉｃｉｐａｌＲｅｆｏｒｍａｎｄＤｅｖｅｌｏｐｍｅｎｔａｎｄＰｒｏｊｅｃｔ（Ｎｏ．２０２１ＺＬ０１２２）． 衍生化⁃气相色谱⁃三重四极杆质谱法测定泼尼松龙中联氨
钱 冲１， 张 梅１，２， 刘珊珊１， 勾新磊１， 王 尉１， 胡光辉１，２∗
（１．北京市理化分析测试中心，有机材料检测技术与质量评价北京市重点实验室
北京１０００８９；２．北京市食品安全分析测试工程技术研究中心，北京１０００８９） 摘要：泼尼松龙是一种广泛用于临床治疗的肾上腺糖皮质激素药物，其中联氨的残留会直接影响用药安全，但目前
国内外还没有出台相应的法律法规和标准来管控药物中联氨的残留限值。联氨具有强极性和强还原性，理化性质
很不稳定，易被氧化，又因缺少发色团，相对分子质量太小，检测起来难度很大，需引入一种衍生化试剂，降低其极
性，生成相对分子质量较大且理化性质稳定的衍生产物。该研究通过优化衍生化试剂、色谱⁃质谱条件、溶剂体系和
衍生化条件，建立了衍生化⁃气相色谱⁃三重四极杆质谱法（ＧＣ⁃ＭＳ／ＭＳ）测定泼尼松龙中联氨残留的方法，并进行了
方法学验证，结果满意。称取１ｇ泼尼松龙样品置于１０ｍＬ具塞离心管中，加入稀释溶剂（甲醇⁃二氯甲烷（１４∶２３，
ｖ／ｖ））至刻度线，涡旋振荡至样品完全溶解后，吸取１００μＬ置于进样小瓶中，再加入丙酮９００μＬ，涡旋振荡混匀，样
品在丙酮⁃稀释溶剂（９∶１，ｖ／ｖ）中同时完成稀释和衍生化反应后，再经ＧＣ⁃ＭＳ／ＭＳ检测分析。该研究的衍生化反
应无需在添加冰乙酸和超声条件下进行，也无需再添加其他试剂进行萃取操作，联氨与丙酮可瞬间发生衍生化反
应，直接实现泼尼松龙中联氨的快速测定。结果表明，联氨在１～１２μｇ／Ｌ范围内线性关系良好，线性相关系数（ｒ２）
为０９９９９；检出限、定量限分别为００３和０１０ｍｇ／ｋｇ；进样精密度（ｒｅｌａｔｉｖｅｓｔａｎｄａｒｄｄｅｖｉａｔｉｏｎ，ＲＳＤ）为１１０％。
加标回收率和重复性良好，加标水平分别为１、６、１２μｇ／Ｌ时的回收率为９６１５％～９６４６％，对应的ＲＳＤ值为１７７％
～２１２％。中间精密度良好，不同时间、不同人员在同一台仪器上测定结果的ＲＳＤ值为１７７％。方法耐用性良好，
通过改变色谱条件来研究检测结果受影响程度大小，在原条件、初始柱温±５℃、升温速率±２℃／ｍｉｎ、柱流量±０１
ｍＬ／ｍｉｎ的条件下分别对加标６μｇ／Ｌ的样品溶液中的联氨含量进行检测，检测结果的ＲＳＤ值为２５８％。应用建
立的方法测定泼尼松龙市售标准品和某药企提供的９个不同批次的泼尼松龙样品，均未检出联氨。该方法操作简
便、准确可靠、灵敏度高、选择性好，可用于泼尼松龙中联氨的检测。
关键词：气相色谱⁃三重四极杆质谱；联氨；泼尼松龙
中图分类号：Ｏ６５８ 文献标识码：Ａ 文章编号：１０００⁃８７１３（２０２１）０７⁃０７５０⁃０８ 文章编号：１０００⁃８７１３（２０２１）０７⁃０７５０⁃０８ 文献标识码：Ａ 中图分类号：Ｏ６５８ 文献标识码：Ａ 文章编号：１０００⁃８７１３（２０２１）０７⁃０７５０⁃０８ ２０２１年７月
Ｊｕｌｙ２０２１ Ｖｏｌ．３９Ｎｏ．７
７５０～７５７ ＣｈｉｎｅｓｅＪｏｕｒｎａｌｏｆＣｈｒｏｍａｔｏｇｒａｐｈｙ 技术与应用 ＤＯＩ：１０．３７２４／ＳＰ．Ｊ．１１２３．２０２１．０３００２ 引用本文：钱冲，张梅，刘珊珊，勾新磊，王尉，胡光辉．衍生化⁃气相色谱⁃三重四极杆质谱法测定泼尼松龙中联氨．色谱，２０２１，３９（７）：
７５０－７５７．
ＱＩＡＮＣｈｏｎｇ，ＺＨＡＮＧＭｅｉ，ＬＩＵＳｈａｎｓｈａｎ，ＧＯＵＸｉｎｌｅｉ，ＷＡＮＧＷｅｉ，ＨＵＧｕａｎｇｈｕｉ．Ｄｅｔｅｒｍｉｎａｔｉｏｎｏｆｈｙｄｒａｚｉｎｅｉｎｐｒｅｄｎｉｓｏｌｏｎｅｂｙ
ｄｅｒｉｖａｔｉｚａｔｉｏｎ⁃ｇａｓｃｈｒｏｍａｔｏｇｒａｐｈｙ⁃ｔｒｉｐｌｅｑｕａｄｒｕｐｏｌｅｍａｓｓｓｐｅｃｔｒｏｍｅｔｒｙ．ＣｈｉｎｅｓｅＪｏｕｒｎａｌｏｆＣｈｒｏｍａｔｏｇｒａｐｈｙ，２０２１，３９（７）：７５０－
７５７． １ 实验部分 联氨具有强极性和强还原性，其理化性质不稳
定，很容易被氧化，在空气中易吸湿冒烟，又因缺少
发色团，相对分子质量太小，检测起来难度很大。目
前联氨常见的检测方法有分光光度法［７］、气相色谱
法（ｇａｓｃｈｒｏｍａｔｏｇｒａｐｈｙ，ＧＣ）［８，９］、气相色谱⁃质谱
法（ｇａｓｃｈｒｏｍａｔｏｇｒａｐｈｙ⁃ｍａｓｓｓｐｅｃｔｒｏｍｅｔｒｙ，ＧＣ⁃
ＭＳ）［１０，１１］、液相色谱法（ｌｉｑｕｉｄｃｈｒｏｍａｔｏｇｒａｐｈｙ，
ＬＣ）［１２，１３］、液相色谱⁃串联质谱法（ｌｉｑｕｉｄｃｈｒｏｍａ⁃
ｔｏｇｒａｐｈｙ⁃ｔａｎｄｅｍ
ｍａｓｓ
ｓｐｅｃｔｒｏｍｅｔｒｙ，
ＬＣ⁃ＭＳ／
ＭＳ）［１４］、离子色谱法（ｉｏｎ
ｃｈｒｏｍａｔｏｇｒａｐｈｙ， Ｄｅｔｅｒｍｉｎａｔｉｏｎｏｆｈｙｄｒａｚｉｎｅｉｎｐｒｅｄｎｉｓｏｌｏｎｅｂｙｄｅｒｉｖａｔｉｚａｔｉｏｎ⁃
ｇａｓｃｈｒｏｍａｔｏｇｒａｐｈｙ⁃ｔｒｉｐｌｅｑｕａｄｒｕｐｏｌｅｍａｓｓｓｐｅｃｔｒｏｍｅｔｒｙ ＱＩＡＮＣｈｏｎｇ１，ＺＨＡＮＧＭｅｉ１，２，ＬＩＵＳｈａｎｓｈａｎ１，ＧＯＵＸｉｎｌｅｉ１，ＷＡＮＧＷｅｉ１，ＨＵＧｕａｎｇｈｕｉ１，２∗
（１．ＢｅｉｊｉｎｇＫｅｙＬａｂｏｒａｔｏｒｙｏｆＯｒｇａｎｉｃＭａｔｅｒｉａｌｓＴｅｓｔｉｎｇＴｅｃｈｎｏｌｏｇｙ＆ＱｕａｌｉｔｙＥｖａｌｕａｔｉｏｎＢｅｉｊｉｎｇＣｅｎｔｒｅ
ｆｏｒＰｈｙｓｉｃａｌａｎｄＣｈｅｍｉｃａｌＡｎａｌｙｓｉｓ，Ｂｅｉｊｉｎｇ１０００８９，Ｃｈｉｎａ；２．ＢｅｉｊｉｎｇＥｎｇｉｎｅｅｒｉｎｇＲｅｓｅａｒｃｈ
ＣｅｎｔｅｒｏｆＦｏｏｄＳａｆｅｔｙＡｎａｌｙｓｉｓ，Ｂｅｉｊｉｎｇ１００８９，Ｃｈｉｎａ） Ａｂｓｔｒａｃｔ：Ｐｒｅｄｎｉｓｏｌｏｎｅｉｓａｎａｄｒｅｎａｌｇｌｕｃｏｃｏｒｔｉｃｏｉｄｄｒｕｇｗｉｔｈｉｍｍｕｎｏｓｕｐｐｒｅｓｓｉｖｅ，ａｎｔｉ⁃
ｉｎｆｌａｍｍａｔｏｒｙ，ａｎｔｉ⁃ａｌｌｅｒｇｉｃ，ａｎｄａｎｔｉｖｉｒａｌｅｆｆｅｃｔｓｔｈａｔａｒｅｗｉｄｅｌｙｅｘｐｌｏｉｔｅｄｉｎｃｌｉｎｉｃａｌｔｒｅａｔｍｅｎｔ．
Ｔｈｅｈｙｄｒａｚｉｎｅｒｅｓｉｄｕｅｔｏｐｒｅｄｎｉｓｏｌｏｎｅｄｉｒｅｃｔｌｙａｆｆｅｃｔｓｍｅｄｉｃａｔｉｏｎｓａｆｅｔｙａｎｄｔｈｒｅａｔｅｎｓｔｈｅ
ｐａｔｉｅｎｔ’ｓｈｅａｌｔｈ．Ａｔｐｒｅｓｅｎｔ，ｔｈｅｒｅａｒｅｎｏｒｅｌｅｖａｎｔｌａｗｓ，ｒｅｇｕｌａｔｉｏｎｓ，ａｎｄｓｔａｎｄａｒｄｓｔｏｃｏｎｔｒｏｌ
ｔｈｅｒｅｓｉｄｕａｌｌｉｍｉｔｏｆｈｙｄｒａｚｉｎｅｉｎｄｒｕｇｓａｔｈｏｍｅｏｒａｂｒｏａｄ．Ｔｈｅｒｅｆｏｒｅ，ａｓｉｍｐｌｅ，ｒａｐｉｄ，ａｃｃｕ⁃
ｒａｔｅ，ｒｅｌｉａｂｌｅ，ｓｅｎｓｉｔｉｖｅ，ａｎｄｓｅｌｅｃｔｉｖｅｍｅｔｈｏｄｉｓｕｒｇｅｎｔｌｙｎｅｅｄｅｄｆｏｒｔｈｅｄｅｔｅｒｍｉｎａｔｉｏｎｏｆｔｒａｃｅ
ｈｙｄｒａｚｉｎｅｉｎｐｒｅｄｎｉｓｏｌｏｎｅ．Ｈｙｄｒａｚｉｎｅｈａｓｓｔｒｏｎｇｐｏｌａｒｉｔｙａｎｄｒｅｄｕｃｔｉｖｉｔｙ，ｗｉｔｈｕｎｓｔａｂｌｅｐｈｙｓｉｃａｌ
ａｎｄｃｈｅｍｉｃａｌｐｒｏｐｅｒｔｉｅｓ，ｔｈｕｓｂｅｉｎｇｅａｓｉｌｙｏｘｉｄｉｚｅｄ．Ｉｎａｄｄｉｔｉｏｎ，ｂｅｃａｕｓｅｏｆｔｈｅｌａｃｋｏｆ 钱 冲，等：衍生化⁃气相色谱⁃三重四极杆质谱法测定泼尼松龙中联氨 第７期 第７期 ·
１
５
７
· ｃｈｒｏｍｏｐｈｏｒｅｓａｎｄｌｏｗｍｏｌｅｃｕｌａｒｗｅｉｇｈｔ，ｔｈｅｄｅｔｅｃｔｉｏｎｏｆｈｙｄｒａｚｉｎｅｉｓｖｅｒｙｄｉｆｆｉｃｕｌｔ．Ｔｈｅｒｅｆｏｒｅ，
ａｄｅｒｉｖａｔｉｖｅｒｅａｇｅｎｔｓｈｏｕｌｄｂｅｉｎｔｒｏｄｕｃｅｄｔｏｒｅｄｕｃｅｉｔｓｐｏｌａｒｉｔｙａｎｄｇｅｎｅｒａｔｅａｄｅｒｉｖａｔｉｖｅｐｒｏｄ⁃
ｕｃｔｗｉｔｈａｈｉｇｈｍｏｌｅｃｕｌａｒｗｅｉｇｈｔａｓｗｅｌｌａｓｓｔａｂｌｅｐｈｙｓｉｃａｌａｎｄｃｈｅｍｉｃａｌｐｒｏｐｅｒｔｉｅｓ．Ａｃｅｔｏｎｅ，
ａｓａｃｏｍｍｏｎｌａｂｏｒａｔｏｒｙｒｅａｇｅｎｔ，ｉｓｉｎｅｘｐｅｎｓｉｖｅａｎｄｃａｎｒａｐｉｄｌｙｒｅａｃｔｗｉｔｈｈｙｄｒａｚｉｎｅ；ｔｈｅｒｅ⁃
ｆｏｒｅ，ｉｔｉｓａｎｉｄｅａｌｄｅｒｉｖａｔｉｖｅｒｅａｇｅｎｔｆｏｒｔｈｅｄｅｔｅｒｍｉｎａｔｉｏｎｏｆｈｙｄｒａｚｉｎｅ．Ｉｎｔｈｉｓｓｔｕｄｙ，ａｍｅｔｈｏｄ
ｂａｓｅｄｏｎｐｒｅｃｏｌｕｍｎｄｅｒｉｖａｔｉｚａｔｉｏｎｗｉｔｈｇａｓｃｈｒｏｍａｔｏｇｒａｐｈｙ⁃ｔｒｉｐｌｅｑｕａｄｒｕｐｏｌｅｍａｓｓｓｐｅｃｔｒｏｍｅ⁃
ｔｒｙ（ＧＣ⁃ＭＳ／ＭＳ）ｗａｓｄｅｖｅｌｏｐｅｄｆｏｒｔｈｅｄｅｔｅｒｍｉｎａｔｉｏｎｏｆｈｙｄｒａｚｉｎｅｉｎｐｒｅｄｎｉｓｏｌｏｎｅｂｙｏｐｔｉｍｉ⁃
ｚｉｎｇｔｈｅｄｅｒｉｖａｔｉｚａｔｉｏｎｒｅａｇｅｎｔ，ＧＣａｎｄＭＳｃｏｎｄｉｔｉｏｎｓ，ｓｏｌｖｅｎｔｓｙｓｔｅｍ，ａｎｄｄｅｒｉｖａｔｉｚａｔｉｏｎｃｏｎ⁃
ｄｉｔｉｏｎｓ．Ｍｅｔｈｏｄｖａｌｉｄａｔｉｏｎｗａｓｔｈｅｎｃａｒｒｉｅｄｏｕｔｕｓｉｎｇｔｈｅｅｓｔａｂｌｉｓｈｅｄｍｅｔｈｏｄ，ａｎｄｔｈｅｒｅｓｕｌｔｓ
ｗｅｒｅｓａｔｉｓｆａｃｔｏｒｙ．Ｉｎｔｈｉｓｓｔｕｄｙ，１ｇｏｆｐｒｅｄｎｉｓｏｌｏｎｅｓａｍｐｌｅｗａｓｗｅｉｇｈｅｄａｎｄｐｌａｃｅｄｉｎａ１０ｍＬ
ｃｅｎｔｒｉｆｕｇｅｔｕｂｅｗｉｔｈａｐｌｕｇ；ｔｈｅｎ，ａｍｅｔｈａｎｏｌ⁃ｄｉｃｈｌｏｒｏｍｅｔｈａｎｅｄｉｌｕｔｉｏｎｓｏｌｖｅｎｔ（１４∶２３，ｖ／ｖ）
ｗａｓａｄｄｅｄｔｏｔｈｅｓｃａｌｅｌｉｎｅ，ａｎｄｔｈｅｓａｍｐｌｅｗａｓｖｏｒｔｅｘｅｄｕｎｔｉｌｃｏｍｐｌｅｔｅｌｙｄｉｓｓｏｌｖｅｄ．Ａｂｏｕｔ１００
μＬｏｆｔｈｅｔｅｓｔｓｏｌｕｔｉｏｎｐｒｅｐａｒｅｄａｂｏｖｅｗａｓｐｉｐｅｔｔｅｄｉｎｔｏｔｈｅｓａｍｐｌｅｖｉａｌ，ｆｏｌｌｏｗｅｄｂｙｔｈｅａｄｄｉ⁃
ｔｉｏｎｏｆ９００μＬａｃｅｔｏｎｅ．Ｔｈｅｒｅｓｕｌｔｉｎｇｓｏｌｕｔｉｏｎｗａｓｖｏｒｔｅｘｅｄａｎｄｍｉｘｅｄｗｅｌｌ．Ｔｈｅｓａｍｐｌｅｗａｓ
ｄｉｌｕｔｅｄａｎｄｄｅｒｉｖａｔｉｚｅｄｓｉｍｕｌｔａｎｅｏｕｓｌｙｉｎａｃｅｔｏｎｅｓｏｌｕｔｉｏｎ，ａｃｅｔｏｎｅ／ｍｅｔｈａｎｏｌ⁃ｄｉｃｈｌｏｒｏｍｅｔｈａｎｅ
ｄｉｌｕｔｉｏｎｓｏｌｖｅｎｔ（９∶１，ｖ／ｖ），ａｎｄｔｈｅｎｄｅｔｅｃｔｅｄａｎｄａｎａｌｙｚｅｄｂｙＧＣ⁃ＭＳ／ＭＳ．Ｉｎｔｈｉｓｓｔｕｄｙ，ｔｈｅ
ｄｅｒｉｖａｔｉｚａｔｉｏｎｒｅａｃｔｉｏｎｂｅｔｗｅｅｎｈｙｄｒａｚｉｎｅａｎｄａｃｅｔｏｎｅｄｉｄｎｏｔｒｅｑｕｉｒｅｔｈｅａｄｄｉｔｉｏｎｏｆａｃｅｔｉｃａｃｉｄ
ａｎｄｕｌｔｒａｓｏｕｎｄｃｏｎｄｉｔｉｏｎｓ，ｏｒｔｈｅｕｓｅｏｆｏｔｈｅｒｒｅａｇｅｎｔｓｆｏｒｔｈｅｅｘｔｒａｃｔｉｏｎｏｐｅｒａｔｉｏｎ．Ｔｈｅｒｅａｃ⁃
ｔｉｏｎｗａｓｉｎｓｔａｎｔａｎｅｏｕｓ，ａｎｄｒａｐｉｄｄｅｔｅｒｍｉｎａｔｉｏｎｏｆｈｙｄｒａｚｉｎｅｉｎｐｒｅｄｎｉｓｏｌｏｎｅｃｏｕｌｄｂｅ
ａｃｈｉｅｖｅｄ．Ｔｈｅｓｔａｎｄａｒｄｃｕｒｖｅｗａｓｏｂｔａｉｎｅｄｗｉｔｈａｇｏｏｄｃｏｒｒｅｌａｔｉｏｎｃｏｅｆｆｉｃｉｅｎｔ（ｒ２＝０９９９９）ｉｎ
ｔｈｅｒａｎｇｅ１－１２μｇ／Ｌ．Ｔｈｅｌｉｍｉｔｓｏｆｄｅｔｅｃｔｉｏｎａｎｄｑｕａｎｔｉｔａｔｉｏｎｗｅｒｅ００３ｍｇ／ｋｇａｎｄ０１０
ｍｇ／ｋｇ，ｒｅｓｐｅｃｔｉｖｅｌｙ．Ｔｈｅｒｅｌａｔｉｖｅｓｔａｎｄａｒｄｄｅｖｉａｔｉｏｎ（ＲＳＤ）ｏｆｉｎｊｅｃｔｉｏｎｐｒｅｃｉｓｉｏｎｗａｓ１１０％．
Ｔｈｅｒｅｃｏｖｅｒｉｅｓａｎｄｒｅｐｅａｔａｂｉｌｉｔｙｗｅｒｅｇｏｏｄ；ｔｈｅｒｅｃｏｖｅｒｉｅｓｏｆｌｏｗ⁃，ｍｅｄｉｕｍ⁃，ａｎｄｈｉｇｈ⁃
ｃｏｎｃｅｎｔｒａｔｉｏｎｓｐｉｋｅｄｓａｍｐｌｅｓｗｅｒｅ９６１５％－９６４６％ａｔｓｐｉｋｅｄｃｏｎｃｅｎｔｒａｔｉｏｎｓｏｆ１，６，ａｎｄ１２
μｇ／Ｌ，ｒｅｓｐｅｃｔｉｖｅｌｙ，ａｎｄｔｈｅｃｏｒｒｅｓｐｏｎｄｉｎｇＲＳＤｓｗｅｒｅ１７７％－２１２％．Ｔｈｅｉｎｔｅｒｍｅｄｉａｔｅｐｒｅｃｉ⁃
ｓｉｏｎｗａｓｇｏｏｄ，ａｎｄｔｈｅＲＳＤｏｆｔｈｅｄｅｔｅｒｍｉｎａｔｉｏｎｒｅｓｕｌｔｓｏｂｔａｉｎｅｄｏｎｔｈｅｓａｍｅｉｎｓｔｒｕｍｅｎｔｂｙ
ｄｉｆｆｅｒｅｎｔｌａｂｏｒａｔｏｒｙｔｅｃｈｎｉｃｉａｎｓａｔｄｉｆｆｅｒｅｎｔｔｉｍｅｓｗａｓ１７７％．Ｔｈｅｄｕｒａｂｉｌｉｔｙｗａｓｇｏｏｄ，ａｎｄｔｈｅ
ｄｅｇｒｅｅｏｆｉｎｆｌｕｅｎｃｅｏｆｔｈｅｄｅｔｅｃｔｉｏｎｒｅｓｕｌｔｓｗａｓｓｔｕｄｉｅｄｂｙｃｈａｎｇｉｎｇｔｈｅｃｈｒｏｍａｔｏｇｒａｐｈｉｃｃｏｎｄｉ⁃
ｔｉｏｎｓ．Ｕｎｄｅｒｔｈｅｏｒｉｇｉｎａｌｃｏｎｄｉｔｉｏｎｏｒｃｏｎｄｉｔｉｏｎｓｗｉｔｈｉｎｉｔｉａｌｃｏｌｕｍｎｔｅｍｐｅｒａｔｕｒｅ±５℃，ｈｅａｔｉｎｇ
ｒａｔｅ±２℃／ｍｉｎ，ｏｒｃｏｌｕｍｎｆｌｏｗｒａｔｅ±０１ｍＬ／ｍｉｎ，ｔｈｅｈｙｄｒａｚｉｎｅｃｏｎｔｅｎｔｉｎｔｈｅｓａｍｐｌｅｓｏｌｕ⁃
ｔｉｏｎａｔａｓｐｉｋｅｄｃｏｎｃｅｎｔｒａｔｉｏｎｏｆ６μｇ／Ｌｗａｓｄｅｔｅｃｔｅｄ，ａｎｄｔｈｅＲＳＤｏｆｔｈｅｄｅｔｅｃｔｉｏｎｒｅｓｕｌｔｓｗａｓ
２５８％．Ｔｈｅｅｓｔａｂｌｉｓｈｅｄｍｅｔｈｏｄｗａｓａｐｐｌｉｅｄｔｏｄｅｔｅｃｔｈｙｄｒａｚｉｎｅｉｎａｐｒｅｄｎｉｓｏｌｏｎｅｓｔａｎｄａｒｄｓｕｂ⁃
ｓｔａｎｃｅｐｒｏｃｕｒｅｄｆｒｏｍｔｈｅｍａｒｋｅｔａｎｄｎｉｎｅｂａｔｃｈｅｓｏｆｐｒｅｄｎｉｓｏｌｏｎｅｓａｍｐｌｅｓｐｒｏｖｉｄｅｄｂｙａｐｈａｒ⁃
ｍａｃｅｕｔｉｃａｌｃｏｍｐａｎｙ．Ｎｏｈｙｄｒａｚｉｎｅｗａｓｄｅｔｅｃｔｅｄｉｎａｎｙｏｆｔｈｅｓｅｓａｍｐｌｅｓ．Ｔｈｅｅｓｔａｂｌｉｓｈｅｄｍｅｔｈ⁃
ｏｄｉｓｓｉｍｐｌｅ，ｒｅｌｉａｂｌｅ，ｈｉｇｈｌｙｓｅｎｓｉｔｉｖｅ，ａｎｄｈｉｇｈｌｙｓｅｌｅｃｔｉｖｅ，ａｎｄｉｔｃａｎｂｅａｐｐｌｉｅｄｆｏｒｔｈｅ
ｄｅｔｅｃｔｉｏｎｏｆｈｙｄｒａｚｉｎｅｉｎｐｒｅｄｎｉｓｏｌｏｎｅ．
Ｋｅｙｗｏｒｄｓ：ｇａｓｃｈｒｏｍａｔｏｇｒａｐｈｙ⁃ｔｒｉｐｌｅｑｕａｄｒｕｐｏｌｅｍａｓｓｓｐｅｃｔｒｏｍｅｔｒｙ（ＧＣ⁃ＭＳ／ＭＳ）；ｈｙｄｒａ⁃
ｚｉｎｅ
ｐｒｅｄｎｉｓｏｌｏｎｅ Ｋｅｙｗｏｒｄｓ：ｇａｓｃｈｒｏｍａｔｏｇｒａｐｈｙ⁃ｔｒｉｐｌｅｑｕａｄｒｕｐｏｌｅｍａｓｓｓｐｅｃｔｒｏｍｅｔｒｙ（ＧＣ⁃ＭＳ／ＭＳ）；ｈｙｄｒａ⁃
ｚｉｎｅ；ｐｒｅｄｎｉｓｏｌｏｎｅ 第３９卷 色 ·
２
５
７
· 谱 ＩＣ）［１５］、荧光分析法［１６］等。上述方法中，分光光度
法灵敏度太低，不能满足低检出限的要求；ＧＣ、ＬＣ、
ＩＣ的基质干扰大，专属性差，复杂基质样品中的其
他组分易干扰其检测分析；ＧＣ⁃ＭＳ的基质效应较
大，重复性不好，累积进样后检测结果偏差较大；
ＬＣ⁃ＭＳ／ＭＳ对实验室条件要求较高，仪器昂贵，检
测成本高，不利于检测方法的普及；荧光分析法需要
构建特定的荧光探针，操作繁琐。相比较而言，气相
色谱⁃三重四极杆质谱法（ｇａｓｃｈｒｏｍａｔｏｇｒａｐｈｙ⁃
ｔｒｉｐｌｅｑｕａｄｒｕｐｏｌｅｍａｓｓｓｐｅｃｔｒｏｍｅｔｒｙ，ＧＣ⁃ＭＳ／
ＭＳ）具有检测成本低、操作简便、灵敏度高、专属性
好、基质干扰小、基质效应低等优点［１７］，目前关于药
物中联氨的ＧＣ⁃ＭＳ／ＭＳ检测方法报道较少，缺乏相
关研究。周延生等［１８］、侯勤勤等［１９］采用丙酮溶液
（每毫升丙酮中加入１μＬ冰乙酸）作为萃取剂和衍
生化试剂，分别对药物和土壤中的联氨进行衍生和
超声提取，再使用ＧＣ氮磷检测器（ｎｉｔｒｏｇｅｎｐｈｏｓ⁃
ｐｈｏｒｕｓｄｅｔｅｃｔｏｒ，ＮＰＤ）对联氨进行测定。该方法
需在添加冰乙酸和超声条件下进行，样品、试剂用量
大，实验操作和ＮＰＤ检测器维护起来繁琐费时。此
外，由于样品处于未溶解状态，会存在联氨提取和衍
生不完全的可能。因为泼尼松龙不能溶于丙酮，本
研究先采用稀释溶剂甲醇⁃二氯甲烷（１４∶２３，ｖ／ｖ）
溶解泼尼松龙，再在丙酮⁃稀释溶剂（９∶１，ｖ／ｖ）中进
行稀释、衍生化，联氨与丙酮的衍生化反应无需在添
加冰乙酸和超声条件下进行，也无需再添加其他试
剂进行萃取操作，联氨与丙酮可瞬间发生衍生化反
应，从而直接实现泼尼松龙中联氨的快速测定。本
文建立了衍生化⁃ＧＣ⁃ＭＳ／ＭＳ检测泼尼松龙中联氨
的方法，并进行了相关的方法学验证，取得了满意的
结果，为泼尼松龙及其他药物中联氨的检测和监控
提供了科学依据和技术支持。 联氨（Ｎ２Ｈ４）是一种无色、油状、易燃的液体，通
常以水合联氨（Ｎ２Ｈ４·Ｈ２Ｏ）的形态存在，是一种重
要的化学试剂，被广泛应用于工业、农业和军事等领
域，在医药、农药、塑料、染料和火箭燃料等行业发挥
着重要作用［１］。联氨具有一种类似氨的刺鼻性气
味，是腐蚀性极强的强碱，对人体危害较大，是一种
刺激性强烈的皮肤致敏物，会损害人体的肝脏、肺、
肾脏、血液和中枢神经系统，国际化学安全项目及综
合风险信息系统根据该系统的证据特征权重给予联
氨Ｂ２的分级（可能的人类致癌物质）［２］。泼尼松龙
（ｐｒｅｄｎｉｓｏｌｏｎｅ）化学名称为１１β，１７α，２１⁃三羟基孕
甾⁃１，４⁃二烯⁃３，２０⁃二酮，是一种广泛用于临床治疗
的肾上腺糖皮质激素药物，具有免疫抑制、抗炎、抗
过敏和抗病毒等功效［３］。盐酸氨基脲是一种重要
的化工中间体，作为医药原料可用于制取硝基呋喃
类等药物，是泼尼松龙合成的重要原料，而水合联氨
又是盐酸氨基脲合成的原料，所以在泼尼松龙的合
成过程中，盐酸氨基脲的使用很有可能会引入联氨
的残留［４］。根据欧洲药事管理局和美国食品药品
管理局的法规要求，基因毒性杂质的毒理学关注阈
值（ｔｈｒｅｓｈｏｌｄｏｆｔｏｘｉｃｏｌｏｇｉｃａｌｃｏｎｃｅｒｎ，ＴＴＣ）限度
为１５μｇ／ｄ，泼尼松龙每天最高服用剂量为２５０
ｍｇ，因此泼尼松龙中联氨的可接受限度为６μｇ／ｇ。
一般药企在生产泼尼松龙时，为了安全起见，当联氨
的残留量低于可接受限度的１０％时，才可以确定泼
尼松龙产品是安全的，所以本研究将泼尼松龙中联
氨的限度定为０６μｇ／ｇ。泼尼松龙中联氨的残留
会直接影响用药安全，威胁人们的身体健康。目前，
国内外还没有出台相应的法律法规和标准来管控药
物中联氨的残留限值［５］。所以，建立一种简便快
捷、准确可靠、灵敏度高、选择性好的泼尼松龙中痕
量联氨的测定方法，对保障临床用药安全具有重要
意义［６］。 １．６ ＧＣ⁃ＭＳ／ＭＳ分析条件 ＧＣ条件：ＶＦ⁃５ＭＳ毛细管色谱柱（３０ｍ×０２５
ｍｍ×０２５μｍ）；进样口温度２５０℃；载气为高纯
Ｈｅ（纯度＞９９９９９％）；分流进样，分流比５∶１；进样量
１０μＬ；恒线速度控制模式；柱流量１０ｍＬ／ｍｉｎ；升
温程序为，初始柱温５０℃保持１ｍｉｎ，２０℃／ｍｉｎ升
至１５０℃，３０℃／ｍｉｎ升至２８０℃，保持５ｍｉｎ。 ＧＣ条件：ＶＦ⁃５ＭＳ毛细管色谱柱（３０ｍ×０２５
ｍｍ×０２５μｍ）；进样口温度２５０℃；载气为高纯
Ｈｅ（纯度＞９９９９９％）；分流进样，分流比５∶１；进样量
１０μＬ；恒线速度控制模式；柱流量１０ｍＬ／ｍｉｎ；升
温程序为，初始柱温５０℃保持１ｍｉｎ，２０℃／ｍｉｎ升
至１５０℃，３０℃／ｍｉｎ升至２８０℃，保持５ｍｉｎ。 ＧＣ条件：ＶＦ⁃５ＭＳ毛细管色谱柱（３０ｍ×０２５
ｍｍ×０２５μｍ）；进样口温度２５０℃；载气为高纯
Ｈｅ（纯度＞９９９９９％）；分流进样，分流比５∶１；进样量
１０μＬ；恒线速度控制模式；柱流量１０ｍＬ／ｍｉｎ；升
温程序为，初始柱温５０℃保持１ｍｉｎ，２０℃／ｍｉｎ升
至１５０℃，３０℃／ｍｉｎ升至２８０℃，保持５ｍｉｎ。 试液，作为供试品溶液。 试液，作为供试品溶液。 １．１ 仪器与试剂 ＧＣＭＳ⁃ＴＱ８０４０气相色谱⁃三重四极杆质谱仪，
ＧＣＭＳｓｏｌｕｔｉｏｎ４３０工作站（日本Ｓｈｉｍａｄｚｕ公司）；
ＸＰＥ１０５电子天平（瑞士ＭｅｔｔｌｅｒＴｏｌｅｄｏ公司）；
Ｖｏｒｔｅｘ⁃Ｇｅｎｉｅ２涡旋振荡器（美国ＳｃｉｅｎｔｉｆｉｃＩｎｄｕｓ⁃
ｔｒｉｅｓ公司）。 甲醇、二氯甲烷、丙酮（色谱纯，美国Ｔｈｅｒｍｏ
ＦｉｓｈｅｒＳｃｉｅｎｔｉｆｉｃ公司）；冰乙酸（分析纯，北京化工
厂）；联氨标准溶液（１０００ｍｇ／Ｌ，国家有色金属及 钱 冲，等：衍生化⁃气相色谱⁃三重四极杆质谱法测定泼尼松龙中联氨 第７期 第７期 ·
３
５
７
· 电子材料分析测试中心国标（北京）检验认证有限
公司）；泼尼松龙标准品（纯度为９９７４％，德国Ｄｒ．
Ｅｈｒｅｎｓｔｏｒｆｅｒ公司）；泼尼松龙样品（某制药公司提
供，用于方法学验证试验）。 ２．１ 衍生试剂的选择 联氨强极性的氨基基团会与毛细管色谱柱的固
定相发生作用，导致色谱峰展宽严重，同时还会引起
严重的柱流失，干扰检测。联氨的相对分子质量小，
如果直接使用ＧＣ⁃ＭＳ／ＭＳ进行检测，易被基质和系
统中的其他碎片离子干扰，同时还会因为生成的母
离子相对分子质量太小，无法二次打碎生成相应的
子离子，无法进行ＧＣ⁃ＭＳ／ＭＳ分析。此外，联氨还
具有强还原性，理化性质不稳定，在检测过程中很容
易发生氧化反应，使检测结果失真，甚至还会出现假
阴性的结果。所以，在联氨的ＧＣ⁃ＭＳ／ＭＳ检测过程
中需引入一种衍生试剂，降低其极性，生成相对分子
质量较大且理化性质稳定的衍生产物。目前文献报
道的衍生化试剂有丙酮［１８，１９］、苯甲醛［２］、对二甲氨
基苯甲醛［７］、氯甲酸乙酯［８］、糠醛［９］、邻苯二甲
醛［１０］、２⁃硝基苯甲醛［１１］、三氟乙酰基丙酮［１７］等，其
中，由于丙酮为实验室常用试剂，且价格低廉，有利
于本方法的普及和推广，因此本研究选择丙酮作为
联氨衍生化试剂，联氨与丙酮的衍生反应方程式如
图１所示。 １．２ 稀释溶剂的配制 １．２ 稀释溶剂的配制
准确量取甲醇１４０ｍＬ、二氯甲烷２３０ｍＬ置于
同一试剂瓶中，摇匀，配制得甲醇⁃二氯甲烷（１４∶２３，
ｖ／ｖ）混合溶液，作为稀释溶剂，在４℃下保存。 １．３ 空白溶液的配制
吸取９００μＬ丙酮置于进样小瓶中，再加入１００
μＬ稀释溶剂，涡旋振荡混匀，作为空白溶剂。 １．４ 标准工作溶液的配制
用丙酮将１０００ｍｇ／Ｌ的联氨标准溶液原液逐
级梯度稀释至１０、２０、４０、６０、１００、１２０μｇ／Ｌ，分别吸
取上述联氨标准溶液各１００μＬ置于不同的进样小
瓶中，再分别加入稀释溶剂１００μＬ和丙酮８００μＬ，
涡旋振荡混匀，即配制得质量浓度分别为１、２、４、６、
１０、１２μｇ／Ｌ的标准工作溶液。所有标准溶液在４
℃下保存。 １．５ 样品处理 称取泼尼松龙样品１ｇ置于１０ｍＬ具塞离心管
中，加入稀释溶剂至刻度线，涡旋振荡至样品完全溶
解，配制得１００ｇ／Ｌ的样品测试液。吸取上述配制
的样品测试液１００μＬ置于进样小瓶中，再加入丙酮
９００μＬ，涡旋振荡混匀，即配制得１０ｇ／Ｌ的样品测
试液作为供试品溶液 图１ 联氨与丙酮的衍生化反应
Ｆｉｇ．１ Ｄｅｒｉｖａｔｉｚａｔｉｏｎｒｅａｃｔｉｏｎｂｅｔｗｅｅｎ
ｈｙｄｒａｚｉｎｅａｎｄａｃｅｔｏｎｅ 图１ 联氨与丙酮的衍生化反应
Ｆｉｇ．１ Ｄｅｒｉｖａｔｉｚａｔｉｏｎｒｅａｃｔｉｏｎｂｅｔｗｅｅｎ
ｈｙｄｒａｚｉｎｅａｎｄａｃｅｔｏｎｅ ２．２ 色谱⁃质谱条件的优化 用丙酮将质量浓度为１０００ｍｇ／Ｌ的联氨标准
溶液原液稀释至１０ｍｇ／Ｌ，然后在ｍ／ｚ６０～１１５范
围内进行全扫描（ｓｃａｎ）分析，依次对丙酮和联氨标
准溶液进行检测。由于丙酮和联氨衍生物（１，２⁃二
（丙烷⁃２⁃亚烷基）肼）的相对分子质量分别为５８和
１１２，所以在该条件下，不仅可以完全消除丙酮以及
基质中ｍ／ｚ大于１１５的碎片离子的干扰，同时还能
满足联氨衍生物的ｓｃａｎ分析。图２为丙酮中干扰
杂质和联氨衍生物总离子流图的叠加图，丙酮中有
干扰杂质与联氨衍生物的保留时间一致，干扰联氨
的检测分析。图３为丙酮杂质和联氨衍生物的质谱
图，比较分析发现ｍ／ｚ１０１的碎片离子同时存在于
丙酮杂质和联氨衍生物的质谱图中，且强度相近， ＭＳ／ＭＳ条件：电子轰击离子源（ＥＩ）；电子能量
７０ｅＶ；离子源温度２３０℃；接口温度２８０℃；溶剂延
迟时间２ｍｉｎ；多重反应监测模式（ｍｕｌｔｉｐｌｅｒｅａｃ⁃
ｔｉｏｎｍｏｎｉｔｏｒｉｎｇ，ＭＲＭ）；联氨衍生物的保留时间为
３５７３ｍｉｎ；定量离子对为ｍ／ｚ１１２００＞９７１０，对应
的碰撞能为６Ｖ；定性离子对为ｍ／ｚ１１２００＞
５６１０、ｍ／ｚ１１２００＞７０１０，对应的碰撞能为１８、 ＭＳ／ＭＳ条件：电子轰击离子源（ＥＩ）；电子能量
７０ｅＶ；离子源温度２３０℃；接口温度２８０℃；溶剂延
迟时间２ｍｉｎ；多重反应监测模式（ｍｕｌｔｉｐｌｅｒｅａｃ⁃
ｔｉｏｎｍｏｎｉｔｏｒｉｎｇ，ＭＲＭ）；联氨衍生物的保留时间为
３５７３ｍｉｎ；定量离子对为ｍ／ｚ１１２００＞９７１０，对应
的碰撞能为６Ｖ；定性离子对为ｍ／ｚ１１２００＞
５６１０、ｍ／ｚ１１２００＞７０１０，对应的碰撞能为１８、 第３９卷 第３９卷 色 ·
４
５
７
· 谱 选取丰度高、质荷比大的特征碎片离子ｍ／ｚ
９７、ｍ／ｚ１１２作为母离子（一级碎片离子），进行二
级质谱分析，对其进行碰撞能优化，对子离子（二级
碎片离子）进行优化选择，选择丰度较高、质荷比较
大、基质干扰较小的离子对进行定性、定量分析。表
１为联氨衍生物的离子对优化结果，由于丙酮中存
在干扰联氨分析的离子对ｍ／ｚ９７００＞５６１０（见图
５），所以本方法舍弃该离子对，选取ｍ／ｚ１１２００＞
９７１０（定量离子对），以及ｍ／ｚ１１２００＞５６１０、
ｍ／ｚ１１２００＞７０１０（定性离子对）对联氨进行检测
分析。图６为该条件下空白溶液和供试品溶液的选
择离子流图，表明空白溶液、供试品溶液中其他组分
不会干扰联氨的检测分析，该方法专属性良好。再
对分流比、升温程序、柱流量、进样口温度、接口温度
等色谱参数进行优化调整，使联氨衍生物的响应和
峰形均达到最佳，缩短方法检测程序的时间，提高检
测效率。联氨衍生物的二级裂解规律如图７所示。
２．３ 溶剂体系的优化 而ｍ／ｚ９７、ｍ／ｚ１１２的碎片离子只存在于联氨衍生
物质谱图中，表明ｍ／ｚ１０１为丙酮基质中干扰杂质
的碎片离子，而ｍ／ｚ９７、ｍ／ｚ１１２为联氨衍生物的
特征碎片离子。联氨衍生物的一级裂解规律如图４
所示。 物质谱图中，表明ｍ／ｚ１０１为丙酮基质中干扰杂质
的碎片离子，而ｍ／ｚ９７、ｍ／ｚ１１２为联氨衍生物的
特征碎片离子。联氨衍生物的一级裂解规律如图４
所示。
图２ 丙酮、联氨衍生物总离子流图的叠加图
Ｆｉｇ．２ Ｏｖｅｒｌａｐｏｆｔｈｅｔｏｔａｌｉｏｎｃｈｒｏｍａｔｏｇｒａｍｓｏｆ
ａｃｅｔｏｎｅａｎｄｈｙｄｒａｚｉｎｅｄｅｒｉｖａｔｉｖｅ
图３ （ａ）丙酮中干扰杂质和（ｂ）联氨衍生物的质谱图
Ｆｉｇ．３ Ｍａｓｓｓｐｅｃｔｒａｏｆ（ａ）ｉｍｐｕｒｉｔｙｉｎｔｅｒｆｅｒｅｎｃｅｉｎ
ａｃｅｔｏｎｅａｎｄ（ｂ）ｈｙｄｒａｚｉｎｅｄｅｒｉｖａｔｉｖｅ 图２ 丙酮、联氨衍生物总离子流图的叠加图
Ｆｉｇ．２ Ｏｖｅｒｌａｐｏｆｔｈｅｔｏｔａｌｉｏｎｃｈｒｏｍａｔｏｇｒａｍｓｏｆ
ａｃｅｔｏｎｅａｎｄｈｙｄｒａｚｉｎｅｄｅｒｉｖａｔｉｖｅ
图３ （ａ）丙酮中干扰杂质和（ｂ）联氨衍生物的质谱图
Ｆｉｇ．３ Ｍａｓｓｓｐｅｃｔｒａｏｆ（ａ）ｉｍｐｕｒｉｔｙｉｎｔｅｒｆｅｒｅｎｃｅｉｎ
ａｃｅｔｏｎｅａｎｄ（ｂ）ｈｙｄｒａｚｉｎｅｄｅｒｉｖａｔｉｖｅ
图
联氨衍生物的
级裂解规律 图２ 丙酮、联氨衍生物总离子流图的叠加图 Ｆｉｇ．２ Ｏｖｅｒｌａｐｏｆｔｈｅｔｏｔａｌｉｏｎｃｈｒｏｍａｔｏｇｒａｍｓｏｆ
ａｃｅｔｏｎｅａｎｄｈｙｄｒａｚｉｎｅｄｅｒｉｖａｔｉｖｅ 平行称取１０份泼尼松龙样品１００ｍｇ于离心管
中，分别加入水、甲醇、乙腈、二氯甲烷、丙酮、Ｎ，Ｎ⁃
二甲基甲酰胺、二甲基亚砜、正己烷、乙酸乙酯、甲苯
各１ｍＬ，充分涡旋振荡，考察样品的溶解情况。发
现除Ｎ，Ｎ⁃二甲基甲酰胺、二甲基亚砜能完全溶解，
乙腈能部分溶解外，其他溶剂的溶解效果均不理想。
同时对联氨衍生物在Ｎ，Ｎ⁃二甲基甲酰胺、二甲基亚 表１ 联氨衍生物的离子对 表１ 联氨衍生物的离子对
Ｔａｂｌｅ１ Ｉｏｎｐａｉｒｓｏｆｈｙｄｒａｚｉｎｅｄｅｒｉｖａｔｉｖｅ
Ｎｏ．
Ｉｏｎｐａｉｒ（ｍ／ｚ）
Ｃｏｌｌｉｓｉｏｎｅｎｅｒｇｙ／Ｖ
Ａｂｕｎｄａｎｃｅ／％
１
１１２．００＞９７．１０
６
１００．００
２
１１２．００＞５６．１０
１８
２７．６８
３
９７．００＞５６．１０
９
２５．８３
４
１１２．００＞７０．１０
９
８．０３
图５ 丙酮的选择离子流图
Ｆｉｇ．５ Ｓｅｌｅｃｔｅｄｉｏｎｃｈｒｏｍａｔｏｇｒａｍｏｆａｃｅｔｏｎｅ 表１ 联氨衍生物的离子对
Ｔａｂｌｅ１ Ｉｏｎｐａｉｒｓｏｆｈｙｄｒａｚｉｎｅｄｅｒｉｖａｔｉｖｅ 表１ 联氨衍生物的离子对
Ｔａｂｌｅ１ Ｉｏｎｐａｉｒｓｏｆｈｙｄｒａｚｉｎｅｄｅｒｉｖａｔｉｖｅ
Ｎｏ．
Ｉｏｎｐａｉｒ（ｍ／ｚ）
Ｃｏｌｌｉｓｉｏｎｅｎｅｒｇｙ／Ｖ
Ａｂｕｎｄａｎｃｅ／％
１
１１２．００＞９７．１０
６
１００．００
２
１１２．００＞５６．１０
１８
２７．６８
３
９７．００＞５６．１０
９
２５．８３
４
１１２．００＞７０．１０
９
８．０３
图５ 丙酮的选择离子流图
Ｆｉｇ．５ Ｓｅｌｅｃｔｅｄｉｏｎｃｈｒｏｍａｔｏｇｒａｍｏｆａｃｅｔｏｎｅ 图３ （ａ）丙酮中干扰杂质和（ｂ）联氨衍生物的质谱图
Ｆｉｇ．３ Ｍａｓｓｓｐｅｃｔｒａｏｆ（ａ）ｉｍｐｕｒｉｔｙｉｎｔｅｒｆｅｒｅｎｃｅｉｎ
ａｃｅｔｏｎｅａｎｄ（ｂ）ｈｙｄｒａｚｉｎｅｄｅｒｉｖａｔｉｖｅ 图３ （ａ）丙酮中干扰杂质和（ｂ）联氨衍生物的质谱图
Ｆｉｇ．３ Ｍａｓｓｓｐｅｃｔｒａｏｆ（ａ）ｉｍｐｕｒｉｔｙｉｎｔｅｒｆｅｒｅｎｃｅｉｎ
ａｃｅｔｏｎｅａｎｄ（ｂ）ｈｙｄｒａｚｉｎｅｄｅｒｉｖａｔｉｖｅ 图３ （ａ）丙酮中干扰杂质和（ｂ）联氨衍生物的质谱图 Ｆｉｇ．３ Ｍａｓｓｓｐｅｃｔｒａｏｆ（ａ）ｉｍｐｕｒｉｔｙｉｎｔｅｒｆｅｒｅｎｃｅｉｎ
ａｃｅｔｏｎｅａｎｄ（ｂ）ｈｙｄｒａｚｉｎｅｄｅｒｉｖａｔｉｖｅ 图４ 联氨衍生物的一级裂解规律 图４ 联氨衍生物的一级裂解规律
Ｆｉｇ．４ Ｆｉｒｓｔｌｅｖｅｌｆｒａｇｍｅｎｔａｔｉｏｎｒｅｇｕｌａｒｉｔｙｏｆ
ｈｙｄｒａｚｉｎｅｄｅｒｉｖａｔｉｖｅ Ｆｉｇ．４ Ｆｉｒｓｔｌｅｖｅｌｆｒａｇｍｅｎｔａｔｉｏｎｒｅｇｕｌａｒｉｔｙｏｆ
ｈｙｄｒａｚｉｎｅｄｅｒｉｖａｔｉｖｅ 图５ 丙酮的选择离子流图 Ｆｉｇ．５ Ｓｅｌｅｃｔｅｄｉｏｎｃｈｒｏｍａｔｏｇｒａｍｏｆａｃｅｔｏｎｅ Ｆｉｇ．５ Ｓｅｌｅｃｔｅｄｉｏｎｃｈｒｏｍａｔｏｇｒａｍｏｆａｃｅｔｏｎｅ Ｆｉｇ．５ Ｓｅｌｅｃｔｅｄｉｏｎｃｈｒｏｍａｔｏｇｒａｍｏｆａｃｅｔｏｎｅ 钱 冲，等：衍生化⁃气相色谱⁃三重四极杆质谱法测定泼尼松龙中联氨 第７期 第７期 ·
５
５
７
· 图６ （ａ）空白溶液和（ｂ）供试品溶液的选择离子流图
Ｆｉｇ．６ Ｓｅｌｅｃｔｅｄｉｏｎｃｈｒｏｍａｔｏｇｒａｍｓｏｆ（ａ）ｔｈｅｂｌａｎｋ
ｓｏｌｕｔｉｏｎａｎｄ（ｂ）ｔｈｅｓａｍｐｌｅｓｏｌｕｔｉｏｎ 醇、二氯甲烷体积比为１４∶２３时，泼尼松龙的溶解度
最好。综上，本研究先采用甲醇⁃二氯甲烷（１４∶２３，
ｖ／ｖ）作为稀释溶剂来溶解泼尼松龙样品，再用丙酮
将其稀释１０倍，即最终的溶剂体系为丙酮⁃稀释溶
剂（９∶１，ｖ／ｖ）。在该溶剂体系下，联氨标准溶液
（６００μｇ／Ｌ）的选择离子流图如图８所示，联氨衍
生物色谱峰响应、峰形均良好。 醇、二氯甲烷体积比为１４∶２３时，泼尼松龙的溶解度
最好。综上，本研究先采用甲醇⁃二氯甲烷（１４∶２３，
ｖ／ｖ）作为稀释溶剂来溶解泼尼松龙样品，再用丙酮
将其稀释１０倍，即最终的溶剂体系为丙酮⁃稀释溶
剂（９∶１，ｖ／ｖ）。在该溶剂体系下，联氨标准溶液
（６００μｇ／Ｌ）的选择离子流图如图８所示，联氨衍
生物色谱峰响应、峰形均良好。 图８ ６００μｇ／Ｌ联氨标准溶液的选择离子流图
Ｆｉｇ．８ Ｓｅｌｅｃｔｅｄｉｏｎｃｈｒｏｍａｔｏｇｒａｍｓｏｆ６００μｇ／Ｌ
ｈｙｄｒａｚｉｎｅｓｔａｎｄａｒｄｓｏｌｕｔｉｏｎ 图８ ６００μｇ／Ｌ联氨标准溶液的选择离子流图 图８ ６００μｇ／Ｌ联氨标准溶液的选择离子流图
Ｆｉｇ．８ Ｓｅｌｅｃｔｅｄｉｏｎｃｈｒｏｍａｔｏｇｒａｍｓｏｆ６００μｇ／Ｌ
ｈｙｄｒａｚｉｎｅｓｔａｎｄａｒｄｓｏｌｕｔｉｏｎ 图６ （ａ）空白溶液和（ｂ）供试品溶液的选择离子流图 图６ （ａ）空白溶液和（ｂ）供试品溶液的选择离子流图
Ｆｉｇ．６ Ｓｅｌｅｃｔｅｄｉｏｎｃｈｒｏｍａｔｏｇｒａｍｓｏｆ（ａ）ｔｈｅｂｌａｎｋ
ｓｏｌｕｔｉｏｎａｎｄ（ｂ）ｔｈｅｓａｍｐｌｅｓｏｌｕｔｉｏｎ Ｆｉｇ．６ Ｓｅｌｅｃｔｅｄｉｏｎｃｈｒｏｍａｔｏｇｒａｍｓｏｆ（ａ）ｔｈｅｂｌａｎｋ
ｓｏｌｕｔｉｏｎａｎｄ（ｂ）ｔｈｅｓａｍｐｌｅｓｏｌｕｔｉｏｎ ２．８ 中间精密度 不同时间、不同人员分别平行配制６份加标水
平为６μｇ／Ｌ的空白加标样品溶液，在同一台仪器上
重复测定，１２次测定的平均加标回收率为９６２３％，
ＲＳＤ值为１７７％，表明本方法的中间精密度良好。
２．９ 方法的耐用性 ２．４ 衍生化条件的优化 文献［１８，１９］报道，当采用丙酮作为衍生化试剂检
测联氨时，需添加冰乙酸和在超声条件下进行衍生
化反应。本研究从超声时间和冰乙酸添加量两方面
对衍生化条件进行优化。首先配制冰乙酸含量为１
ｍＬ／Ｌ的丙酮溶液，再用该溶液按１４节方法配制
质量浓度为６００μｇ／Ｌ的联氨标准溶液，将该溶液
平行分装成８份置于进样小瓶中，分别超声处理０、
１、２、５、１０、１５、３０、６０ｍｉｎ后，分析联氨标准溶液定
量离子对ｍ／ｚ１１２００＞９７１０峰面积的变化趋势
（见图９ａ）。超声时间在０～６０ｍｉｎ范围内时，联氨
标准溶液的峰面积稳定，联氨的衍生反应与超声时
间的长短没有关系。分别配制冰乙酸质量浓度为
０、１、２、５、１０、２５ｍＬ／Ｌ的丙酮溶液，再按１４节方法
分别配制６００μｇ／Ｌ的联氨标准溶液，不同冰乙酸
浓度体系下，联氨标准溶液定量离子对ｍ／ｚ１１２００
＞９７１０峰面积的变化趋势如图９ｂ所示。当冰乙酸
质量浓度为０～５ｍＬ／Ｌ时，联氨标准溶液峰面积稳
定；当冰乙酸浓度为５～２５ｍＬ／Ｌ时，随着冰乙酸浓
度的增加，联氨标准溶液峰面积有逐渐减小的趋势，
表明联氨的衍生化反应不需要在添加冰乙酸的条件
下进行。上述结果表明，联氨与丙酮混合后可直接
发生衍生化反应，该反应不需要添加冰乙酸，也不需
要在超声条件下进行。 图７ 联氨衍生物的二级裂解规律
Ｆｉｇ．７ Ｓｅｃｏｎｄｌｅｖｅｌｆｒａｇｍｅｎｔａｔｉｏｎｒｅｇｕｌａｒｉｔｙｏｆ
ｈｙｄｒａｚｉｎｅｄｅｒｉｖａｔｉｖｅ 图７ 联氨衍生物的二级裂解规律 Ｆｉｇ．７ Ｓｅｃｏｎｄｌｅｖｅｌｆｒａｇｍｅｎｔａｔｉｏｎｒｅｇｕｌａｒｉｔｙｏｆ
ｈｙｄｒａｚｉｎｅｄｅｒｉｖａｔｉｖｅ 砜、乙腈体系中的响应情况进行考察，发现联氨衍生
物在乙腈体系中的色谱峰严重前延，在Ｎ，Ｎ⁃二甲基
甲酰胺、二甲基亚砜体系中响应很低，且峰形不佳。 上述实验结果表明，使用单一溶剂难以同时实
现样品溶解和联氨衍生物响应良好，因此，本研究尝
试使用混合溶剂体系来解决样品的溶解和目标物质
响应的问题。研究发现，采用甲醇⁃二氯甲烷混合溶
剂时，泼尼松龙的溶解有很大改善。甲醇、二氯甲烷
体积比在１１∶２９至２２∶１８范围内时，１００ｍｇ泼尼松
龙能完全溶于１ｍＬ甲醇⁃二氯甲烷混合溶剂；当甲 第３９卷 第３９卷 ·
６
５
７
· 色 谱 续测定６次，以联氨衍生物定量离子对ｍ／ｚ１１２００
＞９７１０峰面积的ＲＳＤ值考察方法的进样精密度。
６次峰面积的ＲＳＤ值为１１０％，表明本方法进样精
密度良好。 图９ （ａ）不同超声时间和（ｂ）乙酸浓度下的联氨标准溶液的
峰面积（ｍ／ｚ１１２．００＞９７．１０）
Ｆｉｇ．９ Ｐｅａｋａｒｅａｓｏｆｈｙｄｒａｚｉｎｅ（ｍ／ｚ１１２．００＞９７．１０）ｉｎ
ｓｔａｎｄａｒｄｓｏｌｕｔｉｏｎｓｗｉｔｈ（ａ）ｄｉｆｆｅｒｅｎｔｕｌｔｒａ⁃
ｓｏｎｉｃｔｉｍｅａｎｄ（ｂ）ａｃｅｔｉｃａｃｉｄｃｏｎｃｅｎｔｒａｔｉｏｎｓ ２．７ 加标回收率和重复性 对泼尼松龙市售标准品和某药企提供的９个不
同批次的泼尼松龙样品进行检测分析，均未检出联
氨残留。配制加标质量浓度分别为１、６、１２μｇ／Ｌ的
低、中、高水平空白加标样品溶液，每个加标水平平
行制备６份样品进行检测。结果如表２所示，不同
加标水平下，联氨的平均回收率为９６１５％～
９６４６％，ＲＳＤ值为１７７％～２１２％，能满足联氨的
检测要求。 表２ 联氨的加标回收率和重复性（ｎ＝６）
Ｔａｂｌｅ２ Ｓｐｉｋｅｄｒｅｃｏｖｅｒｉｅｓａｎｄｒｅｐｅａｔａｂｉｌｉｔｙｏｆ
ｈｙｄｒａｚｉｎｅ（ｎ＝６）
Ａｎａｌｙｔｅ
１μｇ／Ｌ
Ｒｅｃｏｖｅｒｙ／
％
ＲＳＤ／
％
６μｇ／Ｌ
Ｒｅｃｏｖｅｒｙ／
％
ＲＳＤ／
％
１２μｇ／Ｌ
Ｒｅｃｏｖｅｒｙ／
％
ＲＳＤ／
％
Ｈｙｄｒａｚｉｎｅ
９６．２８
２．１２
９６．１５
１．８４
９６．４６
１．７７
２．８ 中间精密度 图９ （ａ）不同超声时间和（ｂ）乙酸浓度下的联氨标准溶液的
峰面积（ｍ／ｚ１１２．００＞９７．１０） Ｆｉｇ．９ Ｐｅａｋａｒｅａｓｏｆｈｙｄｒａｚｉｎｅ（ｍ／ｚ１１２．００＞９７．１０）ｉｎ
ｓｔａｎｄａｒｄｓｏｌｕｔｉｏｎｓｗｉｔｈ（ａ）ｄｉｆｆｅｒｅｎｔｕｌｔｒａ⁃
ｓｏｎｉｃｔｉｍｅａｎｄ（ｂ）ａｃｅｔｉｃａｃｉｄｃｏｎｃｅｎｔｒａｔｉｏｎｓ 钱 冲，等：衍生化⁃气相色谱⁃三重四极杆质谱法测定泼尼松龙中联氨 ·
７
５
７
· ［１１］ ＳｕｂｒａｍａｎｉａｎＳ，ＮａｒａｙａｎａｓａｓｔｒｉＳ，ＲｅｄｄｙＡＲＫ．Ａｎａｌｙｓｔ，
２０１５，１４０（１）：３３０ ２．５ 方法检出限、定量限和线性范围 用优化好的方法依次分析按照１４节配制的标
准工作溶液，以联氨衍生物定量离子对ｍ／ｚ１１２００
＞９７１０的峰面积（ｙ）和联氨质量浓度（ｘ，μｇ／Ｌ）进
行线性回归，联氨在１～１２μｇ／Ｌ范围内线性关系良
好，得到线性回归方程ｙ＝８１１４１ｘ＋５６４３和线性相
关系数ｒ２＝０９９９９。分别以１０倍信噪比（Ｓ／Ｎ）和
３倍Ｓ／Ｎ确定联氨的方法定量限和检出限为０１０
和００３ｍｇ／ｋｇ。结果表明本方法适用于泼尼松龙
中痕量联氨的检测分析。 本方法通过改变色谱条件来研究检测结果受影
响的程度。分别在原条件、初始柱温±５℃、升温速
率±２℃／ｍｉｎ、柱流量±０１ｍＬ的条件下检测６００
μｇ／Ｌ的联氨标准溶液和６μｇ／Ｌ的空白加标样品溶
液，外标法单点计算加标回收样品溶液中联氨的含
量，结果如表３所示。联氨检测结果的ＲＳＤ值为
２５８％，表明本方法耐用性良好，色谱条件的适当改
变不会对检测结果造成太大影响。 表３ 联氨检测方法的耐用性
Ｔａｂｌｅ３ Ｄｕｒａｂｉｌｉｔｙｏｆｈｙｄｒａｚｉｎｅｄｅｔｅｒｍｉｎａｔｉｏｎ
Ａｎａｌｙｔｅ
Ｃ／（μｇ／Ｌ）
Ｏｒｉｇｉｎａｌ
ｃｏｎｄｉｔｉｏｎ
Δ（ｉｎｉｔｉａｌｃｏｌｕｍｎｔｅｍｐｅｒａｔｕｒｅ）／℃
＋５
－５
Δ（ｈｅａｔｉｎｇｒａｔｅ）／（℃／ｍｉｎ）
＋２
－２
Δ（ｃｏｌｕｍｎｆｌｏｗｒａｔｅ）／（ｍＬ／ｍｉｎ）
＋０．１
－０．１
ＲＳＤ／
％
Ｈｙｄｒａｚｉｎｅ
５．８４
５．６３
５．８９
５．９１
５．７２
５．７８
５．５０
２．５８
３ 结论
本研究建立了衍生化⁃ＧＣ⁃ＭＳ／ＭＳ测定泼尼松
龙中联氨的分析方法。样品先经甲醇⁃二氯甲烷（１４
∶２３，ｖ／ｖ）溶解，再经丙酮稀释、衍生后，直接上机检
测分析。该方法操作简便、专属性强、检出限低、准
确度高、基质干扰小、基质效应低、耐用性好，能够满
足泼尼松龙中联氨的检测要求，为泼尼松龙及其他
药物中联氨的检测及监控提供了科学依据和技术
支持。 第７期 第７期 钱 冲，等：衍生化⁃气相色谱⁃三重四极杆质谱法测定泼尼松龙中联氨 参考文献： 参考文献： ［１２］ ＫｏｓｙａｋｏｖＤＳ，ＰｉｋｏｖｓｋｏｉＩＩ，Ｕｌ’ｙａｎｏｖｓｋｉｉＮＶ，ｅｔａｌ．ＩｎｔＪ
ＥｎｖｉｒｏｎＡｎＣｈ，２０１７，９７（４）：３１３ ［１］ ＳｍｏｌｅｎｋｏｖＡＤ，ＳｈｐｉｇｕｎＯＡ．Ｔａｌａｎｔａ，２０１２，１０２：９３ ［２］ ＷａｎｇＲＪ，ＺｈｕＪＤ，ＺｈａｎｇＬＦ，ｅｔａｌ．ＣｈｉｎｅｓｅＪｏｕｒｎａｌｏｆ
ＶｅｔｅｒｉｎａｒｙＤｒｕｇ，２０１７，５１（１１）：５９
王汝金，朱建德，张丽芳，等．中国兽药杂质，２０１７，５１（１１）：
５９ ［１３］ ＳｍｏｌｅｎｋｏｖＡＤ，ＣｈｅｒｎｏｂｒｏｖｋｉｎａＡＶ，ＳｍｉｒｎｏｖＲＳ，ｅｔａｌ．
ＩｎｔＪＥｎｖｉｒｏｎＡｎＣｈ，２０１３，９３（１２）：１２８６ ＩｎｔＪＥｎｖｉｒｏｎＡｎＣｈ，２０１３，９３（１２）：１２８６ ［１４］ ＡｎＺ，ＬｉＰ，ＺｈａｎｇＸ，ｅｔａｌ．ＪＬｉｑＣｈｒｏｍａｔｏｇｒＲＴ，２０１４，３７
（９）：１２１２ ［３］ ＺｈａｎｇＸ．［ＭＳＤｉｓｓｅｒｔａｔｉｏｎ］．Ｔｉａｎｊｉｎ：ＴｉａｎｊｉｎＵｎｉｖｅｒｓｉｔｙｏｆ
ＳｃｉｅｎｃｅａｎｄＴｅｃｈｎｏｌｏｇｙ，２０１０
张旭．［硕士学位论文］．天津：天津科技大学，２０１０ ［３］ ＺｈａｎｇＸ．［ＭＳＤｉｓｓｅｒｔａｔｉｏｎ］．Ｔｉａｎｊｉｎ：ＴｉａｎｊｉｎＵｎｉｖｅｒｓｉｔｙｏｆ
ＳｃｉｅｎｃｅａｎｄＴｅｃｈｎｏｌｏｇｙ，２０１０ ［１５］ ＷａｎｇＺ，ＪｉａｎｇＺＢ，ＬｉＲＹ．ＣｈｉｎｅｓｅＪｏｕｒｎａｌｏｆＣｈｒｏｍａｔｏｇ⁃
ｒａｐｈｙ，２０１６，３４（１０）：９７２
王真，姜振邦，李仁用．色谱，２０１６，３４（１０）：９７２ 张旭．［硕士学位论文］．天津：天津科技大学，２０１０ ［４］ ＫｈａｎＭ，ＫｕｍａｒＳ，ＪａｙａｓｒｅｅＫ，ｅｔａｌ．Ｃｈｒｏｍａｔｏｇｒａｐｈｉａ，
２０１３，７６（１３／１４）：８０１ ［１６］ ＬｉＸＹ．［ＭＳＤｉｓｓｅｒｔａｔｉｏｎ］．Ｂａｏｄｉｎｇ：ＨｅｉｂｅｉＵｎｉｖｅｒｓｉｔｙ，
２０１９
李雪艳．［硕士学位论文］．保定：河北大学，２０１９ ［５］ ＧｉｏｎｆｒｉｄｄｏＥ，ＮａｃｃａｒａｔｏＡ，ＳｉｎｄｏｎａＧ，ｅｔａｌ．ＡｎａｌＣｈｉｍ
Ａｃｔａ，２０１４，８３５：３７ ［１７］ ＯｈＪＡ，ＳｈｉｎＨＳ．ＡｎａｌＣｈｉｍＡｃｔａ，２０１７，９５０：５７ ［６］ ＳｍｏｌｅｎｋｏｖＡＤ．ＲｅｖＪＣｈｅｍ，２０１２，２（４）：３２９ ［１８］ ＺｈｏｕＹＳ，ＬｉＳＹ，ＨａｎＤＳ，ｅｔａｌ．ＣｈｉｎｅｓｅＪｏｕｒｎａｌｏｆＡｎａｌ⁃
ｙｓｉｓＬａｂｏｒａｔｏｒｙ，２００８，２７（３）：８４
周延生，李赛钰，韩东升，等．分析试验室，２００８，２７（３）：８４ ［７］ ＸｉｎｇＳＣ，ＹａｎｇＹ，ＹｕｅＹＰ．ＡｐｐｌｉｅｄＣｈｅｍｉｃａｌＩｎｄｕｓｔｒｙ，
２０１９，４８（１０）：２５２１
邢书才，杨永，岳亚萍．应用化工，２０１９，４８（１０）：２５２１ ［１９］ ＨｏｕＱＱ，ＬｉｕＺＪ，ＸｕＦ，ｅｔａｌ．ＣｈｅｍｉｃａｌＰｒｏｐｅｌｌａｎｔｓａｎｄ
ＰｏｌｙｍｅｒｉｃＭａｔｅｒｉａｌｓ，２０１７，１５（６）：９０
侯倩倩，刘志娟，许峰，等．化学推进剂与高分子材料，
２０１７，１５（６）：９０ ［８］ ＫｈｕｈａｗａｒＭＹ，ＺａｒｄａｒｉＬＡ．ＡｎａｌＳｃｉ，２００８，２４（１１）：１４９３ ［９］ ＨｕＰ，ＺｈａｏＳＭ．ＥａｒｔｈａｎｄＥｎｖｉｒｏｎｍｅｎｔ，２０１７，４５（２）：２４２
胡平，赵世民．地球与环境，２０１７，４５（２）：２４２ ［１０］ ＯｈＪＡ，ＰａｒｋＪＨ，ＳｈｉｎＨＳ．ＡｎａｌＣｈｉｍＡｃｔａ，２０１３，７６９：
７９ ７９
|
https://openalex.org/W4281964614
|
https://zenodo.org/records/6591286/files/%D0%A4.%D0%9C.%D0%A2%D0%B0%D0%B6%D0%B8%D0%BC%D0%B0%D1%82%D0%BE%D0%B2%D0%B0%20.pdf
|
Russian
| null |
ЕР УЧАСТКАСИНИ ОЛДИ-СОТДИ ҚИЛИШНИНГ ЎЗИГА ХОС ХУСУСИЯТЛАРИ
|
Zenodo (CERN European Organization for Nuclear Research)
| 2,022
|
cc-by
| 2,238
|
28 Гражданское право (Особенная часть). Учебник. (автор этой главы Х.Р.Рахманкулов). –Ташкент: ТГЮИ, 2009
-158 с. 27 Раҳмонқулов Ҳ.Р. Олди-сотди шартномаси. –Тошкент: Адолат, 2000. -128 б. Ҳ
р
,
право (Особенная часть). Учебник. (автор этой главы Х.Р.Рахманкулов). –Ташкент: ТГЮИ, 2009 ЕР УЧАСТКАСИНИ ОЛДИ-СОТДИ ҚИЛИШНИНГ ЎЗИГА ХОС
ХУСУСИЯТЛАРИ https://doi.org/10.5281/zenodo.6591286 Ф.М.Тажиматова
ТДЮУ Ихтисослаштирилган филиали
1-босқич талабаси
У.И.Шоназаров
Давлат-ҳуқуқий фанлар
кафедраси мудири в.в.б. раҳбарлигида Аннотация: мақолада муаллиф ер участкасини олди-сотди қилишнинг
ўзига хос хусусиятларини, бу борадаги қонунчилик ва назарий манбаларни таҳлил
қилган. Шунингдек, айрим хорижий сивилист олимларнинг асарларига ҳам
мурожаатлар қилгани ҳолда қиёсий таҳлил қилган. Каит
сўзлар:
ер
участкаси,
олди-сотди,
шартнома,
олди-сотди
шартномаси, кўчмас мулк, мулкдор. Ер инсон, ҳайвонот ва ўсимлик дунёси яшаши учун асос ҳисобланади. Шу
сабабдан, ер бойликларига оқилона муносабатда бўлиш, ундан табиат ва жамият
қонуниятлари талабларига риоя қилган ҳолда фойдаланиш, уни муҳофаза
қилиш ҳар қандай жамиятда асосий масалалардан бири бўлиб келган. Бунда ер
участкаларини фуқаролар ва юридик шахсларга уй-жой қуриш, томорқа
хўжалигини юритиш ёки тадбиркорлик фаолиятини амалга ошириш мақсадида
сотиш алоҳида долзарблик касб этади. Айниқса, Ўзбекистон Республикаси
Президентининг 2017 йил 7 февралдаги ПФ-4947-сон Фармони билан
тасдиқланган
“2017
–
2021
йилларда
Ўзбекистон
Республикасини
ривожлантиришнинг
бешта
устувор
йўналиши
бўйича
Ҳаракатлар
стратегияси”да
ер
участкаларини
тадбиркорлик
субъектларига
бериш
тартибини соддалаштириш масалаларининг белгиланиши ҳамда Ўзбекистон
Республикаси Президентининг 2018 йил 11 октябрдаги “Давлат мулки
объектларини
ва
ер
участкаларига
бўлган
ҳуқуқларни
тадбиркорлик
субъектларига сотиш тартибини соддалаштириш бўйича қўшимча чора-
тадбирлар тўғрисида”ги ПФ-5552-сон Фармонининг қабул қилиниши ер
участкалари ва уларга бўлган ҳуқуқларни сотиш муносабатларини тартибга
солишда янги босқични бошлаб берди. ~ 213 ~ www.interonconf.com www.interonconf.com 27 Раҳмонқулов Ҳ.Р. Олди-сотди шартномаси. –Тошкент: Адолат, 2000. -128 б.
28 Ҳ.Р. Олди-сотди шартномаси. –Тошкент: Адолат, 2000. -128 б.
право (Особенная часть). Учебник. (автор этой главы Х.Р.Рахманкулов). –Ташкент: ТГЮИ, 2009 Фуқаролик ҳуқуқи. Иккинчи қисм. (ушбу боб муаллифи О.Оқюлов). -Тошкент: Илм-Зиѐ, 2008. -6 б Фуқаролик ҳуқуқи. Иккинчи қисм. (ушбу боб муаллифи О.Оқюлов). Тошкент: Илм Зиѐ, 200
30 Брагинский М.И., Витрянский В.В. Договорное право. Ч.2. -М.: Статут, 2007. -196 с. THEORY AND ANALYTICAL ASPECTS OF RECENT RESEARCH
International scientific-online conference
Part 1, Issue 5: MAY 31st 2022 THEORY AND ANALYTICAL ASPECTS OF RECENT RESEARCH
International scientific-online conference
Part 1, Issue 5: MAY 31st 2022 Кўчмас мулкни сотиш шартномасида мазкур шартнома бўйича сотиб
олувчига берилаётган кўчмас мулкни аниқ белгилашга имкон берувчи
маълумотлар,
шу
жумладан,
кўчмас
мулкнинг
муайян
ер
майдонида
жойлашганлигини ёки бошқа бир кўчмас мулкнинг таркибида эканлигини
аниқловчи маълумотлар кўрсатилган бўлиши керак. Ушбу маълумотлар мавжуд
бўлмаса,
кўчмас
мулкни
сотиш
шартномасидаги
тегишли
шартлари
келишилмаган, шартноманинг ўзи эса тузилмаган ҳисобланади. Кўчмас мулкни
сотиш шартномасига кўра, сотиб олувчига шу кўчмас мулкка нисбатан мулк
ҳуқуқи билан бирга, мазкур кўчмас мулк жойлашган ва ундан фойдаланиш учун
зарур бўлган ер майдонининг муайян қисмига бўлган ҳуқуқлар ҳам
топширилади. Сотувчи сотилаётган кўчмас мулк жойлашган ер майдонининг
мулкдори бўлган тақдирда, сотиб олувчига ер майдонининг тегишли қисмига
нисбатан мулк ҳуқуқи, ёки ижара ҳуқуқи, ёки кўчмас мулкни сотиш
шартномасида назарда тутилган бошқа ҳуқуқ берилади. Кўчмас мулкка эгалик ҳуқуқининг бошқа шахсга ўтганлиги давлат
рўйхатидан ўтмагунча сотиб олувчи, ушбу мулк унинг эгалигида бўлишидан ёки
ундан фойдаланишидан қатъий назар ушбу мулкни тасарруф этиш ҳуқуқига эга
бўлмайди.27 Бундай ҳаракатларга йўл қўйса, иккинчи тараф олди-сотди
шартномасини ҳақиқий эмас деб топиш учун судда даъво қўзғатиш ҳуқуқига эга. Кўчмас мулкнинг тўлиқ қиймати тўлаб бўлинганидан ҳамда Давлат мулки
қўмитаси, унинг ҳудудий бошқармаси томонидан давлат варақаси берилгандан
кейин ҳаридор мулк эгаси ҳуқуқига эга бўлади. Ер участкаси кўчмас мулк сифатида фуқаролик муомаласида бўлиш
лаёқатига эга. “Ер участкасини фуқаролик муомаласида бўлиши деганда,
универсал ҳуқуқий ворислик тартибида (мерос қилиб қолдириш, юридик
шахсни қайта ташкил этиш), ёҳуд бошқа йўллар билан бир шахсдан бошқасига
эркин олиб бериш ёки ўтказиш имконияти тушунилади”. Юридик адабиётларда ер участкаси олди-сотди шартномаси тушунчаси
берилган бўлиб, унга кўра “олди-сотди шартномаси бўйича бир томон (сотувчи)
бошқа томонга (сотиб олувчига) ер участкасини мулк қилиб бериш
мажбуриятини олади, сотиб олувчи эса ер участкасини олиш ва маълум суммани
тўлаш мажбуриятини олади”. Ҳ.Р.Раҳмонқуловнинг фикрича, кўчмас мулкни сотиш шартномаси олди-
сотди шартномасининг мустақил тури ҳисобланади. Шу сабабли унга олди-
сотди шартномаси тушунчасини аниқловчи барча белгилар хосдир, яъни
мулкнинг ўтиши, икки томонлилик, ҳақ эвазигалик ва консенсуаллик28. ~ 214 ~ www.interonconf.com ~ 215 ~
р
,
р
Д
р
р
у , 31 Мирзаабдуллаева М.Р. Ерга бўлган ҳуқуқни вужудга келишида танловнинг аҳамияти // Ер қонунчилигини
такомиллаштириш муаммолари: республика илмий-амалий конференция материаллари. -Тошкент: ТДЮИ,
2005. –Б.82-84. www.interonconf.com THEORY AND ANALYTICAL ASPECTS OF RECENT RESEARCH
International scientific-online conference
Part 1, Issue 5: MAY 31st 2022 О.Оқюлов ҳам кўчмас мулкни сотиш шартномасини олди-сотди шартномасини
предмети бўйича алоҳида тури эканлигини таъкидлайди29. М.И.Брагинский
ва
В.В.Витрянскийлар
ҳам
кўчмас
мулкни
сотиш
шартномаси олди-сотди шартномасининг предметига кўра алоҳида тури
эканлигини эътироф этишади30. Демак, ердан фойдаланиш ҳуқуқини сотиш шартномаси қўйидаги
белгилари билан ифодаланади: Демак, ердан фойдаланиш ҳуқуқини сотиш шартномаси қўйидаги
белгилари билан ифодаланади: 1. Ердан фойдаланишга оид олди-сотди шартномаси икки томонлама
шартномалар туркумига киради. Шартнома юзасидан тарафларнинг ҳар қайсида
муайян ҳуқуқ ва тегишли мажбурият пайдо бўлади. 2. Ердан фойдаланишга оид олди-сотди шартномаси консенсуал хусусиятга
эга, чунки тузилган шартнома билан уни ижро этиш муддатлари ҳар доим ҳам
бир-бирига мос келавермайди. 3. Ердан фойдаланишга оид олди-сотди шартномаси ҳақ эвазига
тузиладиган шартномалардандир. Сотиб олувчи ер участкасини олиш ва маълум
суммани тўлаш мажбуриятини олади. 4. Ердан фойдаланишга оид олди-сотди шартномасини тузишдан
кўзланган
асосий
мақсад-сотувчи
ер
участкасини
мулк
қилиб
бериш
мажбуриятини олади. 4. Ердан фойдаланишга оид олди-сотди шартномасини тузишдан
кўзланган
асосий
мақсад-сотувчи
ер
участкасини
мулк
қилиб
бериш
мажбуриятини олади. Ер участкасини сотиш жараёнида аукционни ўтказиш тартиби муҳим
босқич бўлиб, аукцион ташкилотчи томонидан таклиф этилган аукционист
ёрдамида ўтказилади. Аввалига объект номи, муайян белгилари, аукцион
босқичлари эълон қилинади. Бу босқич объект баҳосининг 5.15 % атрофида
бўлади. Савдо жараёнида аукционист муайян босқич суммасини эълон қилади,
сотиб олишга рози бўлган иштирокчилар эса ўз билетини кўтаради. Савдо
бошланғич
нарх
эълон
қилинганидан
сўнг
бошланади. Нарх
эълон
қилинганидан кейин билет кўтарган шахс эълон қилинади. Шундан сўнг
аукционист навбатдаги нархни эълон қилади. Савдо якунида эса аукционист
объектнинг сотилганлиги, нархини, харидорнинг билет рақамини эълон
қилади. Савдо якунланганидан сўнг баённома тузилади. Баённомада ер
участкаси жойлашган жой, туман, шаҳар, охирги баҳо, харидор номи қайд
этилади. Баённома харидор (аукцион ғолиби) ва аукцион ташкилотчиси
томонидан имзоланиб, унинг бир нусхаси харидорга берилади. Бу баённома ер
участкасига умрбод эгалик қилиш ҳужжати ҳисобланади. Агар ғолиб чиққан
харидор ўн кун муддат ичида ер участкасига бўлган ҳуқуқни ўтказиш
тўғрисидаги шартномага имзо чекмаса, аукцион баённомаси бекор қилинади. ~ 215 ~ www.interonconf.com www.interonconf.com THEORY AND ANALYTICAL ASPECTS OF RECENT RESEARCH
International scientific-online conference
Part 1, Issue 5: MAY 31st 2022 мажбуриятлар белгилаган холда ер участкаларини бериш ва бўш турган давлат
кўчмас мулк объектларини сотиш тўғрисида қарорлар қабул қилиш. Бунда,
Қорақалпоғистон Республикаси Вазирлар Кенгаши раиси ва вилоятлар
ҳокимларининг мазкур масаладаги қарорлари дастлаб Қорақалпоғистон
Республикаси Жўқорғи Кенгеси ва халқ депутатлари вилоятлар кенгашларида
кўриб чиқилиши ҳамда тасдиқланиши лозим; в) «E-IJRO AUKSION» электрон савдо майдончасида аукционга чиқариш
учун ер участкаларининг жойлашган ҳудудидан келиб чиқиб, Қорақалпоғистон
Республикаси Молия вазирлиги ва вилоят ҳокимликларининг молия бош
бошқармалари, Қорақалпоғистон Республикаси ва вилоятлар давлат солиқ
бошқармалари, Қорақалпоғистон Республикаси ва вилоятлар ер ресурслари ва
давлат кадастри бошқармаларининг тавсияларига асосан, ҳар бир туман бўйича
бўш ер участкаларидан фойдаланиш ҳуқуқининг (0,01 га майдони учун)
бошланғич баҳосини ҳар чоракда тасдиқлаш; в) «E-IJRO AUKSION» электрон савдо майдончасида аукционга чиқариш
учун ер участкаларининг жойлашган ҳудудидан келиб чиқиб, Қорақалпоғистон
Республикаси Молия вазирлиги ва вилоят ҳокимликларининг молия бош
бошқармалари, Қорақалпоғистон Республикаси ва вилоятлар давлат солиқ
бошқармалари, Қорақалпоғистон Республикаси ва вилоятлар ер ресурслари ва
давлат кадастри бошқармаларининг тавсияларига асосан, ҳар бир туман бўйича
бўш ер участкаларидан фойдаланиш ҳуқуқининг (0,01 га майдони учун)
бошланғич баҳосини ҳар чоракда тасдиқлаш; г) уч йилдан ортиқ муддат давомида тупроқ бонитировкаси ишлари олиб
борилмаган уч гектаргача бўлган майдонга эга ер участкаларида тегишли
маҳаллий бюджетлар маблағлари ҳисобига тупроқ бонитировкаси ишлари
буюртмачиси бўлиш; д) «E-IJRO AUKSION» электрон савдо майдончасида аукцион орқали
суғорилмайдиган ерлар тоифасига кирадиган ер участкаларидан вақтинча
фойдаланиш
ҳуқуқини
тадбиркорлик
ва
шаҳарсозлик
фаолияти
учун
инвестициявий (камида 1 млн. АҚШ доллари эквивалентида) ва ижтимоий
мажбуриятлар белгилаган ҳолда 50 йил муддатга сотиш; д) «E-IJRO AUKSION» электрон савдо майдончасида аукцион орқали
суғорилмайдиган ерлар тоифасига кирадиган ер участкаларидан вақтинча
фойдаланиш
ҳуқуқини
тадбиркорлик
ва
шаҳарсозлик
фаолияти
учун
инвестициявий (камида 1 млн. АҚШ доллари эквивалентида) ва ижтимоий
мажбуриятлар белгилаган ҳолда 50 йил муддатга сотиш; е) «E-IJRO AUKSION» электрон савдо майдончасида аукцион орқали
халқаро ва давлат аҳамиятидаги автомобиль йўлларига туташ ҳудудлардаги ер
участкаларидан доимий фойдаланиш ҳуқуқини уларнинг тоифаларидан қатъий
назар Қорақалпоғистон Республикаси Вазирлар Кенгаши ва вилоятлар
ҳокимликлари томонидан тасдиқланган намунавий лойиҳалар бўйича йўлбўйи
ва туристик инфратузилма объектларини ташкил этиш учун сотиш. Бунда,
суғориладиган қишлоқ хўжалиги ерлари тоифасига кирувчи ер участкалари
кўрсатилган мақсадлар учун фақат Ўзбекистон Республикаси Президентининг
2018 йил 11 сентябрдаги ПҚ-3939-сон қарорига биноан ташкил этилган
Тадбиркорликни қўллаб-қувватлаш ва ривожлантириш бўйича комиссия билан
келишилган ҳолда берилади; е) «E-IJRO AUKSION» электрон савдо майдончасида аукцион орқали
халқаро ва давлат аҳамиятидаги автомобиль йўлларига туташ ҳудудлардаги ер
участкаларидан доимий фойдаланиш ҳуқуқини уларнинг тоифаларидан қатъий
назар Қорақалпоғистон Республикаси Вазирлар Кенгаши ва вилоятлар
ҳокимликлари томонидан тасдиқланган намунавий лойиҳалар бўйича йўлбўйи
ва туристик инфратузилма объектларини ташкил этиш учун сотиш. THEORY AND ANALYTICAL ASPECTS OF RECENT RESEARCH
International scientific-online conference
Part 1, Issue 5: MAY 31st 2022 Аукцион якуни тўғрисидаги маълумот 30 календар куни ичида матбуот ёки
махсус информацион бюллетенда эълон қилинади31. Аукцион якуни тўғрисидаги маълумот 30 календар куни ичида матбуот ёки
махсус информацион бюллетенда эълон қилинади31. Аукцион натижаларини расмийлаштириш босқичида биринчи навбатда,
аукцион якуни ҳақидаги баённома имзолангандан сўнг ўн кун ичида туман ёки
шаҳар ҳокимияти ва ер участкасини сотиб олган шахс ўртасида ер участкасига
мерос қилиб қолдириладиган умрбод эгалик қилиш ҳуқуқини берувчи
шартнома имзоланади. Шундан сўнг белгиланган муддат ҳамда тўлов шаклига
риоя қилган ҳолда ер участкасини сотиб олган шахс ҳокимият ҳисобига
келишилган пул миқдорини тўлиқ ҳолда ўтказиши лозим. Аукцион якуни
тўғрисидаги баённомага имзо қўйилганидан кейин 30 календар куни ичида
аукционнинг бошқа иштирокчиларига гаров пуллари қайтариб берилади. Туман
ёки шаҳар ҳокимияти томонидан ер участкасига мерос қилиб қолдириладиган
умрбод эгалик қилиш ҳуқуқини берувчи давлат ҳужжати берилганидан сўнг
ерга эгалик ҳуқуқи давлатдан ерни аукционда сотиб олган шахсга ўтади. У ер
участкасини сотиб юбориш (тасарруф этиш)дан бошқа барча ҳуқуқларга эга
бўлади. Ер участкасига нисбатан мулк ҳуқуқи давлатда сақланиб қолади. Ер
участкаларини фуқароларга умрбод эгалик қилишга сотишдан тушган
маблағларнинг 5 фоизи аукцион ташкилотчисига, 60 фоизи унинг филиалига, 40
фоизи Республика кўчмас мулк биржасига ўтказилади. Ер участкасини сотишдан
тушган пулнинг 20 фоизи Республика бюджетига, 75 фоизи Қорақалпоғистон
Республикаси, вилоятлар ва Тошкент шаҳар бюджетига, ушбу сумманинг 45
фоизи вилоятларнинг шаҳар, туман бюджетларига ўтказилади. Республика ҳудудларида инвестиция муҳитини тубдан яхшилаш,
тадбиркорлик фаолиятини амалга ошириш учун фойдаланилмаётган давлат
мулки объектлари ва ер участкаларига бўлган ҳуқуқларни тадбиркорлик
субъектларига сотишни жадаллаштириш мақсадида: а) ўз ваколати доирасида (жумладан, Қорақалпоғистон Республикаси
Жўқорғи Кенгеси ва вилоятлар халқ депутатлари кенгаши қарорларига асосан),
бўш турган давлат кўчмас мулк объектларини (мазкур Фармоннинг 4-
бандидакўрсатилган объектлар бундан мустасно) уларни баҳоламасдан, 1 сўмга
тенг қийматдаги бошланғич нархда «E-IJRO AUKSION» электрон савдо
майдончасида аукционга чиқариш орқали инвестициявий ва ижтимоий
мажбуриятларни белгилаган ҳолда ўрнатилган тартибда сотиш; б) ўз ваколатлари доирасида инвестиявий лойиҳаларни амалга ошириш
учун аукцион ўтказмасдан, инвестор билан тўғридан-тўғри музокаралар орқали
инвестициявий (камида 1 млн. АҚШ доллари эквивалентида) ва ижтимоий ~ 216 ~ www.interonconf.com THEORY AND ANALYTICAL ASPECTS OF RECENT RESEARCH
International scientific-online conference
Part 1, Issue 5: MAY 31st 2022 Бунда,
суғориладиган қишлоқ хўжалиги ерлари тоифасига кирувчи ер участкалари
кўрсатилган мақсадлар учун фақат Ўзбекистон Республикаси Президентининг
2018 йил 11 сентябрдаги ПҚ-3939-сон қарорига биноан ташкил этилган
Тадбиркорликни қўллаб-қувватлаш ва ривожлантириш бўйича комиссия билан
келишилган ҳолда берилади; ж) ички ишлар органлари ва Миллий гвардиянинг жамоат тартибини
сақлаш кучларининг уч сменали режимдаги патрулини таъминлаш орқали,
шунингдек, мазкур зоналарда сайёҳлар учун дам олишни ташкил этиш
дастурларини тасдиқлаган ҳолда Нукус шаҳри, вилоятлар марказлари ва
сайёҳлар ташриф буюрадиган бошқа жойларда савдо-кўнгилочар ва умумий ~ 217 ~ овқатланиш
объектлари
фаолиятини
тунги
вақтда
чеклашга
қўйилмайдиган зоналарни аниқлаш. овқатланиш
объектлари
фаолиятини
тунги
вақтда
чеклашга
й
қўйилмайдиган зоналарни аниқлаш. Ў йўл Ўзбекистон Республикаси Президентининг 2019 йил 10 январдаги
“Урбанизация
жараёнларини
тубдан
такомиллаштириш
чора-тадбирлари
тўғрисида”ги ПФ-5623-сон Фармонида белгиланишича, 2019 йил 1 июлдан
бошлаб шундай тартиб ўрнатилсинки, унга мувофиқ: юридик шахслар — Ўзбекистон Республикаси резидентлари уларга мулк
ҳуқуқи асосида тегишли бўлган ёки улар томонидан хусусийлаштирилаётган
бино ва иншоотлар, саноат инфратузилмаси объектлари жойлашган ер
участкаларини, шунингдек уларга туташ ҳудудлардаги ишлаб чиқариш
фаолиятини амалга ошириш учун зарур миқдордаги ер участкаларини
хусусийлаштириш ҳуқуқига эга; жисмоний шахслар — Ўзбекистон Республикаси фуқаролари уларга якка
тартибда уй-жой қуриш ва турар жойга хизмат кўрсатиш учун ажратилган ер
участкаларини хусусийлаштириш ҳуқуқига эга; жисмоний ва юридик шахслар — Ўзбекистон Республикаси резидентлари
томонидан хусусийлаштирилган ер участкалари хусусий мулк (фуқаролик
муомаласи объектлари) ҳисобланади ва дахлсиздир, шунингдек Ўзбекистон
Республикасининг
«Хусусий
мулкни
ҳимоя
қилиш
ва
мулкдорлар
ҳуқуқларининг
кафолатлари
тўғрисида»ги Қонунига
мувофиқ
давлат
томонидан муҳофаза қилинади; ер участкаларини хусусийлаштириш фақатгина кадастр ҳужжатлари
мавжуд бўлган ҳолда ҳамда Ўзбекистон Республикаси Вазирлар Маҳкамаси
томонидан белгиланадиган миқдорларда пуллик асосда амалга оширилади. Хулоса
сифатида
айтганда,
ер
участкасини
олди-сотди
қилиш
шартномасини фуқаролик-ҳуқуқий тартибга солиш асослари бугунги кунда
тубдан ўзгариш арафасида эканлигини таъкидлаб ўтиш лозим. Чунки, ер
участкаларининг хусусийлаштирилиши натижасида ер участкалари фуқаролик
муомаласининг мустақил объекти сифатида муомалага киритилиши мумкин
бўлади. Бу эса ўз навбатида ФК ва ЕКнинг янги таҳрири қабул қилинишига олиб
келиши билан бирга, ер участкаларини мулк ҳуқуқи билан бирга сотишнинг
ҳуқуқий механизмларини ишлаб чиқиш заруратини кўрсатади. ФОЙДАЛАНИЛГАН АДАБИЁТЛАР РЎЙХАТИ 1. Раҳмонқулов Ҳ.Р. Олди-сотди шартномаси. –Тошкент: Адолат, 2000. -
128 б. Раҳмонқулов Ҳ.Р. Олди-сотди шартномаси. –Тошкент: Адолат, 2000. 2. Гражданское право (Особенная часть). Учебник. (автор этой главы
Х.Р.Рахманкулов). –Ташкент: ТГЮИ, 2009 -158 с. 2. Гражданское право (Особенная часть). Учебник. (автор этой главы
Х.Р.Рахманкулов). –Ташкент: ТГЮИ, 2009 -158 с. ~ 218 ~ THEORY AND ANALYTICAL ASPECTS OF RECENT RESEARCH 3. Фуқаролик ҳуқуқи. Иккинчи қисм. (ушбу боб муаллифи О.Оқюлов). -
Тошкент: Илм-Зиё, 2008. -6 б. 4. Брагинский М.И., Витрянский В.В. Договорное право. Ч.2. -М.: Статут,
2007. -196 с. 5. Мирзаабдуллаева М.Р. Ерга бўлган ҳуқуқни вужудга келишида
танловнинг аҳамияти // Ер қонунчилигини такомиллаштириш муаммолари:
республика илмий-амалий конференция материаллари. -Тошкент: ТДЮИ, 2005. –Б.82-84. 5. Мирзаабдуллаева М.Р. Ерга бўлган ҳуқуқни вужудга келишида
танловнинг аҳамияти // Ер қонунчилигини такомиллаштириш муаммолари:
республика илмий-амалий конференция материаллари. -Тошкент: ТДЮИ, 2005. –Б.82-84. ~ 219 ~ www.interonconf.com www.interonconf.com
|
https://openalex.org/W2175293256
|
https://www.econstor.eu/bitstream/10419/146840/1/841192774.pdf
|
English
| null |
Cultural insights of CSI: how do Italian and Iranian firms differ?
|
Journal of innovation and entrepreneurship
| 2,015
|
cc-by
| 6,425
|
Journal of Innovation and Entrepreneurship Provided in Cooperation with:
Springer Nature Springer Nature Suggested Citation: Canestrino, Rossella; Bonfanti, Angelo; Oliaee, Leila (2015) : Cultural insights
of CSI: How do Italian and Iranian firms differ?, Journal of Innovation and Entrepreneurship, ISSN
2192-5372, Springer, Heidelberg, Vol. 4, pp. 1-9,
https://doi.org/10.1186/s13731-015-0026-4 Terms of use: Die Dokumente auf EconStor dürfen zu eigenen wissenschaftlichen
Zwecken und zum Privatgebrauch gespeichert und kopiert werden. Documents in EconStor may be saved and copied for your personal
and scholarly purposes. Sie dürfen die Dokumente nicht für öffentliche oder kommerzielle
Zwecke vervielfältigen, öffentlich ausstellen, öffentlich zugänglich
machen, vertreiben oder anderweitig nutzen. You are not to copy documents for public or commercial purposes, to
exhibit the documents publicly, to make them publicly available on the
internet, or to distribute or otherwise use the documents in public. If the documents have been made available under an Open Content
Licence (especially Creative Commons Licences), you may exercise
further usage rights as specified in the indicated licence. Sofern die Verfasser die Dokumente unter Open-Content-Lizenzen
(insbesondere CC-Lizenzen) zur Verfügung gestellt haben sollten,
gelten abweichend von diesen Nutzungsbedingungen die in der dort
genannten Lizenz gewährten Nutzungsrechte. http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ http://creativecommon Canestrino, Rossella; Bonfanti, Angelo; Oliaee, Leila
Article
Cultural insights of CSI: How do Italian and Iranian firms
differ?
Journal of Innovation and Entrepreneurship
Provided in Cooperation with:
Springer Nature
Suggested Citation: Canestrino, Rossella; Bonfanti, Angelo; Oliaee, Leila (2015) : Cultural insights
of CSI: How do Italian and Iranian firms differ?, Journal of Innovation and Entrepreneurship, ISSN
2192-5372, Springer, Heidelberg, Vol. 4, pp. 1-9,
https://doi.org/10.1186/s13731-015-0026-4 Canestrino, Rossella; Bonfanti, Angelo; Oliaee, Leila
Article
Cultural insights of CSI: How do Italian and Iranian firms
differ? Journal of Innovation and Entrepreneurship
Provided in Cooperation with:
Springer Nature
Suggested Citation: Canestrino, Rossella; Bonfanti, Angelo; Oliaee, Leila (2015) : Cultural insights
of CSI: How do Italian and Iranian firms differ?, Journal of Innovation and Entrepreneurship, ISSN
2192-5372, Springer, Heidelberg, Vol. 4, pp. 1-9,
https://doi.org/10.1186/s13731-015-0026-4 * Correspondence:
rossella.canestrino@uniparthenope.it
1Department of Managerial Studies
and Quantitative Methods,
Parthenope University, Via Generale
Parisi 13, Naples 80132, Italy
Full list of author information is
available at the end of the article RESEARCH Open Access © 2015 Canestrino et al. Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in
any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons
license, and indicate if changes were made. Rossella Canestrino1*, Angelo Bonfanti2 and Leila Oliaee3 Rossella Canestrino1*, Angelo Bonfanti2 and Leila Oliaee3 Rossella Canestrino1*, Angelo Bonfanti2 and Leila Oliaee3 * Correspondence:
rossella.canestrino@uniparthenope.it
1Department of Managerial Studies
and Quantitative Methods,
Parthenope University, Via Generale
Parisi 13, Naples 80132, Italy
Full list of author information is
available at the end of the article Abstract Both scholars and practitioners have paid little attention to the social innovation
issue at for-profit firm’s level. This is particularly true with reference to the role that
culture and cultural diversities may have in shaping firms’ orientation toward Corporate
Social Innovation (CSI). According to the underlined observation, this paper aims to fill
the existing research gap about the topic. Through an inductive approach, an extensive
literary review about the cultural drivers of innovation and Corporate Social Responsibility
(CSR) has been carried to gain a wide understanding of CSI. Hofstede’s cultural model
has been, then, used to interpret the cultural insights of CSI in two geographical and
psychological distant countries, namely Italy and Iran. This cross-cultural comparison will
allow both academics and practitioners to recognize culture as key driver of firms’
propensity to adopt social innovative practices. Keywords: Innovation, Corporate social responsibility, Corporate social innovation,
Culture, Cultural diversity, Italian and Iranian firms, Hofstede’s model Canestrino et al. Journal of Innovation and Entrepreneurship (2015) 4:12
DOI 10.1186/s13731-015-0026-4 Background Adopting a socially responsible behavior means
that firms’ innovative actions are oriented toward socially valuable goals, thus CSI be-
longs to both SI and CSR framework. In other words, managing CSI demands to evalu-
ate both firms’ innovation activities and their social responsibility in an integrated
manner. Additionally, if firms’ innovation may be considered the output of a knowledge
management process (Nonaka and Takeuchi 1995; Galunic and Rodan, 1998; Apanaso-
vich 2014), CSI may be interpreted as a way to manage firms’ knowledge in order to
get not only profits but also social goals. Its effectiveness depends on the firms’ ability
to manage both knowledge for innovation and their propensity to achieve social
benefits. As widely highlighted in management literature, culture affects both firms’ innova-
tiveness and CSR (e.g., Kaasa and Vadi 2010). Since values and beliefs differently shape
firms’ innovative behavior, culture may enable the emergence of CSI. However, firms’
involvement in social innovative practices may differ according to the cultural values
that prevail in a given context. Taking into account these considerations, we aim to
investigate the role of culture and cultural diversities, in defining CSI. The paper proceeds as follows. First, we highlight the linkage between culture,
innovation, and CSR by discussing that culture plays in defining firms’ innovative be-
havior as well as firms’ propensity to devote its actions to social aims. Next, we discuss
the concept of CSI and the use Hofstede’s cultural model to interpret the cultural in-
sights of CSI in two geographical and psychological different countries, namely Italy
and Iran. Conclusions , research limits and the undertaken research method are pro-
posed at the end. Firm’s innovation attitude and social responsible orientation: how does culture matter? Culture plays a very important role in both the innovation field and the ground of so-
cial responsibility. Focusing on innovation, many authors examined the linkage between
national culture (Hofstede 1980; Kaasa and Vadi 2010; Petrakis and Pantelis 2014) and
firms’ propensity to innovate, by mainly concentrating on the initiation stage of the
process that means on the creativity stage (e.g., Parjanen 2012). More frequently,
scholars investigate the role of Hofstede’s cultural dimensions (power distance, indi-
vidualism versus collectivism, masculinity versus femininity, uncertainty avoidance, and
long-term orientation as the fifth dimension added to the original model) on national
innovation rates (e.g., Kaasa and Vadi 2010; Taylor and Wilson 2012). Background The concept of social innovation (SI) has been one of the most discussed in the field
of innovation for the last years (e.g., Ashta et al. 2014) and even getting stronger in the
discussion of social development (Rüede and Lurtz 2012). According to Mulgan
(2006), SI refers to innovative activities and services that are motivated by the goal of
meeting a social need and that are predominantly developed through organizations
whose primary purpose is social. SI refers to ideas, concepts, processes, and outcomes
that address social needs in new ways and, if truly successful, fundamentally change so-
cial, economic, or other relations (Hansson et al. 2014). For example, the introduction
of innovative approaches to help disable people, the emergence of online self-help
health groups, the use of distance learning systems to develop knowledge (Maggioni
and Del Giudice 2011; Del Giudice et al. 2013), as well the adoption of technology to
manage the food problems (Bonfanti et al. 2013) in underdeveloped countries and
environmental pollution are all examples of SI practices. In addition to governments, educational institutions, associations (i.e., fair trade),
and user movements (i.e., Wikipedia, Linux), individual entrepreneurs and commercial
corporations (Krull 2008; Baccarani and Golinelli, 2013) may also develop SI. Despite the increasing amount of literature about SI, scholars have paid little atten-
tion to the adoption of its practices at corporate level that means to the development
of Corporate Social Innovation (CSI) (Tham 2010). The term has been articulated by Canestrino et al. Journal of Innovation and Entrepreneurship (2015) 4:12 Page 2 of 9 Page 2 of 9 Page 2 of 9 Rosabeth Moss Kanter (1999) for the first time. The author argued that firms should
use social issues as a learning laboratory for identifying unmet needs and developing
solutions. According to the given perspective, CSI may be defined as a way of finding
new products and services that meet not only the functional needs of consumers but
also their wider aspirations as citizens (Webb 2007). CSI is hitherto under-explored research area because of problems about both its un-
derstanding and its management. On the one hand, understanding CSI requires exam-
ining the existing overlapping between the theoretical backgrounds usually used to
explain these concepts: firms’ innovation theories, on one side, and corporate social re-
sponsibility (CSR), on the other one. Background The analysis of
these studies allows us to point out the two following considerations: (a) each cultural
dimension differently affects the different stages of an innovation process (innovation Canestrino et al. Journal of Innovation and Entrepreneurship (2015) 4:12 Page 3 of 9 Page 3 of 9 creation and innovation transfer and diffusion), by shaping the way new knowledge is
created and then diffused, and (b) a general agreement seems to prevail in literature
about the positive relationship between individualism, masculinity, power distance,
long-term orientation, and firms’ propensity to create innovation. The linkage between culture and CSR is widely discussed in the business ethics field
(e.g. Husted 2001; Robertson 2002). Within the referred research arena, many scholars
recognize that what is ethically right or wrong is culturally affected. But, what does
culture mean? Culture may be considered as the set of driving rules and core beliefs that develop
in a society over time (Schein 1985) and these drive both individuals and firms’
choices. In line with this notion, and taking into account the mainstream literature
about the topic, culture and cultural diversities may affect the following: (a) the as-
sumption about what may be considered moral or not in different countries (Husted
and Allen 2008; Moores 2003); (b) firms’ propensity to adopt a CSR behavior; and (c)
firms’ practicing CSR (explicit vs implicit). Research in CSR has identified remarkable
differences between companies from different countries (Canestrino et al. 2012). For
example, Matten and Moon (2008) pointed out that US corporations usually explicitly
commit to CSR, whereas European business responsibility tends to be more implicit. In coherence with our literary examination, we agree with the following three aspects:
(a) individualism positively relates to firms’ propensity to adhere and to adopt “explicit”
social responsible practices. The higher the individualism index is, the more corpo-
rates’ policies, programs, and strategy could devote to the stakeholders interests. The
lower individualism is, the higher is the adoption of “implicit” responsible practices;
(b) masculinity and power distance negatively relate to firms’ propensity to adhere
and to adopt social responsible practices. The higher masculinity (or power distance)
index is, the less corporates’ policies, programs, and strategy devote to the stake-
holders interests; and (c) uncertainty avoidance positively relates to firms’ propensity
to adhere and to adopt social responsible practices. Background The higher the uncertainty avoid-
ance is, the more corporates’ policies, programs, and strategy could devote to the
stakeholders interests. Crossing the main findings of our theoretical investigation, we examine CSI in Italy
and Iran and highlight cultural differences for CSI between these two countries in the
next sections. Understanding CSI: a comparison between Italy and Iran Putting together the examined literary contributions, we define CSI as the introduction
of a new way, method, or system (innovation) to meet collective needs in a social respon-
sible manner. Our definition enlarges the previous notions of CSI by introducing the
need for a socially responsible orientation not only referring to the aimed goals but also
with respect to the way the goals (societal needs) are satisfied. Therefore, CSI requires
both a high firm’s propensity to innovate and a high CSR orientation. With reference to
both these aspects, Italy and Iran seem to contrast. In the field of innovative activities, the Global Innovation Index (GII) provides rele-
vant data to compare the countries’ innovation dynamics at the global level. It ranks
the innovation performance of 143 countries and economies around the world, based
on 81 indicators. Canestrino et al. Journal of Innovation and Entrepreneurship (2015) 4:12 Canestrino et al. Journal of Innovation and Entrepreneurship (2015) 4:12 Page 4 of 9 Page 4 of 9 Page 4 of 9 According the Global Innovation Index (2014), Italy scores for 45.65 and ranks for 31
within the global countries’ classification. Conversely, Iran locates at 120th position at
global level (Table 1). Italy and Iran significantly differ in their innovative performance at global level, con-
firming an innovative divide between the two countries. The innovation divide between
the two countries reduces with reference to the income group classification. This sug-
gests us to consider the national income as a moderating factor in the influence that
cultural diversities have on local innovative behavior. A national index does not exist for the evaluation of CSR. The most suitable tool to com-
pare firms’ social orientation in a wider context is, therefore, the Global Entrepreneurship
Monitor (GEM) that documents the prevalence of social entrepreneurship (SE) in a given
population. In this regard, the management literature does not provide uniform definition
or understanding of SE (e.g., Bacq and Janssen 2011). For example, SE includes activities of
individuals and groups (social entrepreneurs) who identify gaps in the social system as an
opportunity to serve groups who are marginalized in different ways and aims to address
these needs in entrepreneurial ways (Hansson et al. 2014). In this sense, SE and SI are
interrelated but not interchangeable concepts. Understanding CSI: a comparison between Italy and Iran GEM uses a broad definition of SE, the
last one interpreted as entrepreneurial activity with a social orientation and that in-
volves an organization of individual (Mair and Martí 2006; Zahra et al. 2009). Social
activity includes the following categories of players: (1) traditional NGO (explicit so-
cial purpose—not self-sustaining—not innovative); (2) not for profit SE (explicit social
purpose—not self-sustaining—innovative); (3) hybrid SE (overlapping social and eco-
nomic purposes—self-sustaining); and (4) for profit SE (explicit economic purpose—-
socially committed self-sustaining). As showed in Table 2, both Italy and Iran rates for strictly defined SE are below the
average for—respectively—Western Europe and Middle and North Africa (MENA). But
while Italy is about 30 % far from its average, Iran diverges only for about 8 %. This
suggests that Iran performs better than Italy within its own geographical area. The
underlined gap reduces when we refer to the broadly defined SE (that includes NGOs
and for profit SE). In the Table 2, while “strictly defined” includes only not-for-profit
SE, and hybrid SE (both economic and socially oriented) as parts of the spectrum,
“broadly defined” includes all the five categories. Obviously, the data need to be justified within each national context. This is because
social enterprises can take numerous forms, are engaged in multiple spheres of activity,
and because legal structures vary from country to country. For example, in Italy, social
enterprises are constrained by a non-distribution clause—that is, all income has to be
reinvested in the enterprise. In Iran, the Government’s absolute monopoly in the econ-
omy is one of the most important factors that prevent the provision of necessary re-
quirements for the implementation of social entrepreneurship. Legally, most of the Table 1 Global innovation index—Italy versus Iran
Country
Score (0–100)
Rank
Income
Rank
Region
Rank
Italy
45.65
31
High Income
30
Europe
20
Iran
26.14
120
Upper middle income
35
Central and Southern Asia
6
Source: our adaptation by Global Innovation Report 2014 Source: our adaptation by Global Innovation Report 2014 Page 5 of 9 Page 5 of 9 Canestrino et al. Understanding CSI: a comparison between Italy and Iran Journal of Innovation and Entrepreneurship (2015) 4:12 Table 2 Rates of social entrepreneurship by types and countries
Country
Traditional
NGO
Not-for-
profit SE
Economically
oriented hybrid SE
Socially
oriented
hybrid SE
For-profit
SE
Strictly
defined SE
Broadly
defined SE
Italy
0.27
0.55
1,32
0.45
0.69
2.32
3.28
Average for
Western Europe
0.27
1.11
1,01
1.19
1.01
3.31
4.58
Percentage on
the average
100
49.5
130
37.8
68
70
71.6
Iran
0.06
0.45
1,25
0.17
0.62
1.87
2.55
Average for MENA
0.14
0.76
0,76
0.51
0.80
2.03
2.97
Percentage on
the average
43
59.2
164
33.3
77.5
92.1
85.8
Source: our adaptation by Global Entrepreneurship Monitor (2009) Adult Population Survey
Bold data symbolizes the average for Western Europe and Iran Table 2 Rates of social entrepreneurship by types and countries Source: our adaptation by Global Entrepreneurship Monitor (2009) Adult Population Survey
Bold data symbolizes the average for Western Europe and Iran rules on social efforts are obsolete or not up-to-date. Politically, social issues are ma-
jorly viewed from the political and security points of view. This could hamper the effect
of non-governmental organizations and pose problems for the engagement of SE
(Razavi et al. 2014). SE may also cover a wide range of organizations from cooperatives to public service
providers, from community/voluntary associations to “work insertion” organizations,
and from companies limited by guarantee, thus posing huge attention in the interpret-
ation of the available data. According to this, using the cultural lens to interpret SE al-
lows us to contextualize data, as well as to give a punctual picture of SE that prevail in
a given country. Interpreting the diversities in CSI between Italy and Iran: a cultural perspective A first cultural comparison between Italy and Iran bases on the diversities in Hofstede’s
rankings, as it is showed in the Table 3. The scale for each dimension runs from 0 to
100 with 50 as a midlevel. The rule of thumb is that if a score is under 50, the culture
scores relatively low on that scale, and if any score is over 50, the culture scores high
on that scale. In the case of IDV—the low side (under 50), is considered “Collectivist”
and above 50 considered “Individualist”. A country with a score of 43 would be collect-
ivist but less collectivist than someone with 28 who is moving down toward the 0 mark
(Hofstede et al. 2010). The cross-cultural comparison between Italy and Iran shows that significant diver-
sities exist between the two countries with reference to individualism, masculinity,
and long-term orientation. In particular, at a score of 76, Italy is an individualistic Table 3 Hofstede’s model: a comparison between Italy and Iran
Cultural dimensions
Italy
Iran
Individualism (IND)
76
41
Power distance (PD)
50
58
Masculinity (MAS)
70
43
Uncertainty avoidance (UA)
75
59
Long-term orientation (LO)
61
14
Source: our adaptation by Hofstede et al. (2010) Table 3 Hofstede’s model: a comparison between Italy and Iran
Cultural dimensions
Italy
Iran
Individualism (IND)
76
41
Power distance (PD)
50
58
Masculinity (MAS)
70
43
Uncertainty avoidance (UA)
75
59
Long-term orientation (LO)
61
14
Source: our adaptation by Hofstede et al. (2010) Source: our adaptation by Hofstede et al. (2010) Canestrino et al. Journal of Innovation and Entrepreneurship (2015) 4:12 Page 6 of 9 Page 6 of 9 Page 6 of 9 and “me”-centered culture. Conversely, Iran, with a score of 41, is considered a col-
lectivistic society. This is manifested in a close long-term commitment to the member
“group,” be that a family, extended family, or extended relationships. At 70, Italy is a
masculine society—highly successful—oriented, and driven. Children are taught from
an early age that competition is good and to be a winner is important in one’s life. As
the working environment is the place where every Italian can reach his/her success,
competition among colleagues for making a career can be very strong. Iran scores 43
on this dimension and is thus considered a relatively feminine society. Interpreting the diversities in CSI between Italy and Iran: a cultural perspective In feminine
countries, the focus is on “working in order to live,” managers strive for consensus;
people value equality, solidarity, and quality in their working lives. Conflicts are re-
solved by compromise and negotiation. Incentives such as free time and flexibility are
favored. Femininity culturally justifies the arising of a more social entrepreneurial
orientation, as it has been discussed above. The mentioned differences could justify the diversities in both the propensity to
innovate and the orientation to social responsible practices that characterize the se-
lected countries. It does not mean that knowing the cultural dimensions that
characterize a given context allows us to predict local CSR, firms’ innovative behavior,
and lastly, CSI. There is no quick fix to understand CSI. But the cultural dimensions let
us to predict a little better what is likely to happen. And they become more useful as
we go from the specific case to the trend, average, or expectation. Since IND positively
relates to firms’ propensity to innovate, as well as to their attitude to adopt “explicit”
social responsible practices, one, for example, would expect that CSI arises in individu-
alistic context more easily than in collectivistic ones or that feminine societies (like
Iran) devote to social aims more than masculine ones (like Italy), thus affecting the way
firms develop their own initiatives. From a cultural perspective, for example, Iran shows high levels of power distance,
collectivism, femininity and uncertainty avoidance, and low levels of long-term orienta-
tion. As data display, Iran characterizes also for low levels of innovation and high levels
of social entrepreneurship. In addition, empirical evidences reveal the involvement of
both firms and NGO in social-oriented activities, which means in SE. Because of the
high collectivism, the group’s interest prevails over the individual one at societal level;
femininity plays, therefore, a very important role in fostering local solidarity. Central to the understanding of Iranian SE is also the strong influence of Islam on all
aspects of socio-economic life. In Arabic countries, Islam is much more than a religion;
it is a way of life, and in many countries, it still has a greater importance than the offi-
cial law. That is the Islamic moral law, even if not applied by government, still influ-
ences people’s way of life (Cone 2003). Interpreting the diversities in CSI between Italy and Iran: a cultural perspective According to the taqwā paradigm (Dusuki 2008), each individual is claimed to make
any deliberate effort to achieve the objectives of Shari’ah in the ways prescribed by
Shari’ah itself. As a consequence, people become voluntarily committed to reach the
central goals of human welfare or falah. Furthermore, it plays a unifying role, binding
the community together and building the ground for equality, solidarity, and freedom. Unfortunately, in spite of the underlined considerations, we cannot say anything else
about the existing linkage between culture and CSR mainly because of the established
relationship between and among the different cultural dimensions as well as between
each of them and firms’ behavior. Page 7 of 9 Page 7 of 9 Canestrino et al. Journal of Innovation and Entrepreneurship (2015) 4:12 Results and Discussion As already underlined, we have supported the idea that CSI is affected by culture. Since no CSI rankings exist at global level, the GII and GEM ranks have been used to
picture the differences between Italy and Iran, as regards to their innovative and social
responsible orientation. Italy performs better than Iran in innovation, but compared to Italy, Iran shows a
higher level of SE, within the MENA. Without neglecting the relevance that many other
dimensions—economical,
political,
and
cultural
environments
(Razavi
et
al. 2014)—have on the development of firms’ social and innovative attitude, the mentioned
diversities have been examined in the field of cultural studies. According to Hofstede’s
model, Iran and Italy significantly differ in the following cultural dimensions: (a) indi-
vidualism: relatively high for Italy and low for Iran and (b) masculinity: relatively high
for Italy and low for Iran. The underlined cultural divide may be—even partially—responsible for firms’ attitude
toward innovation, SE, and the adoption of CSI, at least. Obviously, there is no quick
fix to understand CSI. But the cultural dimensions let us to predict a little better what
is likely to happen. Since individualism positively relates to firms’ propensity to
innovate, as well as to their attitude to adopt “explicit” social responsible practices, one,
for example, could expect that CSI arises in individualistic context more easily than in
collectivistic ones. Similar considerations could be developed referring to femininity,
since feminine societies (like Iran) devote to social aims more than masculine ones (like
Italy). Depending on the above, individualistic and feminine contexts would provide the
best background for the emergence of CSI. Despite this, we cannot say anything else, mainly because the relationship that estab-
lish between and among the different cultural dimensions, as well as between each of
them and firms’ propensity to innovate and to devote to social practices. Conclusions This paper provides new insights in the field of CSI by adopting a cultural perspective. The concept of CSI has been examined taking into account the two literary fields to
which it belongs to, namely innovation and CSR. Within the reviewed literature,
scholars agree on the existing linkage between firms' innovative behaviour and culture
on one hand, as well as between the adoption of social responsible practices and the
values that prevail in a given context, on the other hand. In spite of our research re-
sults, however, many challenges also derive from the effect that cultural norms have on
the definition of SE and CSI. Cross-country comparisons of SE and CSI often become
difficult “because everyone speaks from their own regionally defined version of the con-
cepts” (Kerlin 2009); in developing countries, every economic activity seems to have a
social component, while in other countries like the USA, social and business enter-
prises are generally seen as different kinds of organizations. Additionally, the driving forces behind social innovations are manifold. Companies
and organizations from all industries (Palacios-Marqués et al. 2015; Trequattrini et al. 2015; Soto-Acosta et al. 2010, 2015; Meroño-Cerdan and Soto-Acosta 2005) and from
different cultural contexts may engage social innovative practices for several reasons. Their behavior may be affected by many factors as well as culture—notably national Canestrino et al. Journal of Innovation and Entrepreneurship (2015) 4:12 Page 8 of 9 Page 8 of 9 Page 8 of 9 wealth, history, personalities, and even coincidences. The above considerations, there-
fore, mirror the main limits of our paper. Our research represents only a little step in the exploration of a wide and often mis-
understood concept. We recognize that many questions are still unanswered. What
does it happen, for example, when high levels of individualism and masculinity cross? How does power distance—or even uncertainty avoidance—influence CSI? Despite its
inner limits, our paper contributes to the literature improving about CSI by developing
a more comprehensive knowledge and culturally based perspective. Competing interests Competing interests
The authors declare that they have no competing interests. Authors’ contributions CR; BA; and OL, carried an investigation about the cultural drivers that lie upon the adoption of CSI, comparing firms'
propensity to adopt innovation and to adhere to social responsible practices in two geographical and cultural
distant countries, namely Italy and Iran. BA explored the insights of firms' innovation attitude and social responsible
orientation; OL provided the picture to understand Iranian firms' behavior; CR developed the cultural interpretation for
the emerging differences between Italian and Iranian firms. All the three authors conceived the study, partecipated in
its design and drafted the manuscript. All the authors read and approved the final manuscript. Received: 14 October 2015 Accepted: 10 November 2015 Received: 14 October 2015 Accepted: 10 November 2015 Received: 14 October 2015 Accepted: 10 November 2015 Methods Our paper is based on an inductive approach. An extensive literary review about the
cultural drivers that lie under the adoption of innovation and corporate social responsi-
bility has been carried to gain a wide understanding of CSI. The examined literary con-
tributions have been then crossed with the available data about the country innovation
dynamics, on one side, and the rates of social entrepreneurship by countires, on the
other side, in order to compare firms'attitude toward SI in two different countires,
namely, Italy and Iran. Hofstede'cultural model has been, finally, used to interpet the
cultural insights of CSI in the two selected countries. References Apanasovich, N. (2014). Modes of innovation: a grounded meta-analysis. J Knowl Econ, 12, 1–18 Apanasovich, N. (2014). Modes of innovation: a grounded meta-analysis. J Knowl Econ, 12, 1–18. microcredit to microfinance. J Innov Entrep, 3(4), 1–23. Baccarani C., Golinelli G.M. (2013) "Per una rivisitazione delle relazioni tra impresa e territorio", Sinergie, n. 84, pp. VII-XIII. Bacq, S., & Janssen, F. (2011). The multiple faces of social entrepreneurship: a review of definitional issues based on
geographical and thematic criteria. Entrep Reg Dev, 23(5–6), 373–403. Baccarani C., Golinelli G.M. (2013) "Per una rivisitazione delle relazioni tra impresa e territorio", Sinergie, n. 84, pp. VII-XIII
Bacq, S., & Janssen, F. (2011). The multiple faces of social entrepreneurship: a review of definitional issues based on
geographical and thematic criteria. Entrep Reg Dev, 23(5–6), 373–403. Bonfanti, A., Brunetti, F., & Castellani, P. (2013). The last minute market model: an i
assortment management in a sustainability perspective. Sinergie, 91, 173–192. Bonfanti, A., Brunetti, F., & Castellani, P. (2013). The last minute market model: an in
assortment management in a sustainability perspective. Sinergie, 91, 173–192. Canestrino, R., Magliocca, P., & Nocilla, G. (2012). Cross-border knowledge transfer in MNCs: the role of international
executives in managing cultural diversities. In AAVV political change, cultural dynamics and competitiveness of firms
(pp. 86–114). Enzo Albano Editore: Napoli. Canestrino, R., Magliocca, P., & Nocilla, G. (2012). Cross-border knowledge transfer in MNCs: the role of international
executives in managing cultural diversities. In AAVV political change, cultural dynamics and competitiveness of firms
(pp. 86–114). Enzo Albano Editore: Napoli. pp
p
Cone, M. H. (2003). Corporate citizenship. The role of commercial organizations in an Islamic society. J Corp Citiz,
9, 49–66. Cone, M. H. (2003). Corporate citizenship. The role of commercial organizations in an Islamic society. J Corp Citiz,
9, 49–66. Del Giudice, M., Della Peruta, M. R., & Maggioni, V. (2013). Collective knowledge and organizational routines within
academic communities of practice: an empirical research on science–entrepreneurs. J Knowl Econ, 4(3), 260–278. Dusuki, A. W. (2008). Why does Islam say about corporate social responsibility. Rev Islam Econ, 12(1), 5–28. Galunic, C., & Rodan, S. (1998). Resource re-combinations in the firm: knowledge structures and the potential for
Schumpeterian innovation. Strateg Manag J, 19(12), 1193–201. Global Innovation Index. (2014). The human factor in innovation, second printing. Fontainebleau, Ithaca, and Geneva:
Cornell University, INSEAD, and WIPO. Hansson, F., Norn, M. T., & Vad, T. B. (2014). Author details
1D
f 1Department of Managerial Studies and Quantitative Methods, Parthenope University, Via Generale Parisi 13, Naples
80132, Italy. 2Department of Business Administration, University of Verona, Via dell’Artigliere 19, Verona 37129, Italy. 3Department of Human Resource, Azad University - Mashad branch, Faculty of Management, 75, Bozorgmehr 5h st.,
Sajjad blv., Mashad 9186753717, Iran. References Modernize the public sector through innovation? A challenge for the role of
applied social science and evaluation. Evaluation, 20(2), 244–260. Del Giudice, M., Della Peruta, M. R., & Maggioni, V. (2013). Collective knowledge and organizational routines within
academic communities of practice: an empirical research on science–entrepreneurs. J Knowl Econ, 4(3), 260–278. Dusuki, A. W. (2008). Why does Islam say about corporate social responsibility. Rev Islam Econ, 12(1), 5–28. Galunic, C., & Rodan, S. (1998). Resource re-combinations in the firm: knowledge structures and the potential for
Schumpeterian innovation. Strateg Manag J, 19(12), 1193–201. Global Innovation Index (2014) The human factor in innovation second printing Fontainebleau Ithaca and Geneva: Schumpeterian innovation. Strateg Manag J, 19(12), 1193–201. Global Innovation Index. (2014). The human factor in innovation, second printing. Fontainebleau, Ithaca, and Geneva:
Cornell University, INSEAD, and WIPO. Global Innovation Index. (2014). The human factor in innovation, second printing. Fontainebleau, Ithaca, and Geneva:
Cornell University, INSEAD, and WIPO. Hansson, F., Norn, M. T., & Vad, T. B. (2014). Modernize the public sector through innovation? A challenge for the role of
applied social science and evaluation. Evaluation, 20(2), 244–260. Hansson, F., Norn, M. T., & Vad, T. B. (2014). Modernize the public sector through innovation? A challenge for the role of
applied social science and evaluation. Evaluation, 20(2), 244–260. Page 9 of 9 Canestrino et al. Journal of Innovation and Entrepreneurship (2015) 4:12 Hofstede, G. (1980). Culture’s consequences: international differences in work-related values. Los Angeles: Sage Publication. Hofstede, G., Hofstede, G. J., & Minkov, M. (2010). Cultures and organizations: software of the mind. USA: McGraw-Hill. g
Husted, B. W., & Allen, D. B. (2008). Toward a model of cross-cultural business ethics: the impact of individualism and Husted, B. W., & Allen, D. B. (2008). Toward a model of cross-cultural business ethics: the impact of individualism an
ll
i i
h
hi
l d
i i
ki
J B
E hi
82(2) 293 305 Husted, B. W., & Allen, D. B. (2008). Toward a model of cross-cultural business ethics: the impact of individua
collectivism on the ethical decision-making process. J Bus Ethics, 82(2), 293–305. p
collectivism on the ethical decision-making process. J Bus Ethics, 82(2), 293–305. collectivism on the ethical decision-making process. J Bus Ethics, 82(2), 293–305. Husted, B. W. (2001). The impact of individualism and collectivism on the ethical decision-making by individuals in
organizations. References Barriers to social entrepreneurship in Iran: an
li
ti
f
d d th
J E t
O
M
3(2) 2 5 Razavi, S. M., Asadi, M., Esfandabadi, H. M., & Ekbatani, H. (2014). Barriers to social entrepreneurship in Iran: an
application of grounded theory. J Entrep Organ Manag, 3(2), 2–5. Robertson, D. C. (2002). Business ethics across cultures. In M. J. Gannon & K. L. Newman (Eds.), The handbook of cross-
cultural management. Oxford: Blakwell. Rüede, D., & Lurtz, K. (2012). Mapping the various meanings of social innovation: towards a differentiated understanding of an emerging concept. EBS Bus Sch Res Pap Ser, 12(3), 1–51. Schein, E. H. (1985). Defining organizational culture. Class Organ Theory, 3, 490–502. Schein, E. H. (1985). Defining organizational culture. Class Organ Theory, 3, 490–502. ,
g
g
g
y, ,
Soto-Acosta, P., Casado-Lumbreras, C., & Cabezas-Isla, F. (2010). Shaping human capital in software development teams:
h
f
bl d b
f oto-Acosta, P., Casado-Lumbreras, C., & Cabezas-Isla, F. (2010). Shaping Soto-Acosta, P., Casado-Lumbreras, C., & Cabezas-Isla, F. (2010). Shaping human capital in software development teams
the case of mentoring enabled by semantics IET Softw 4(6) 445–452 Soto-Acosta, P., Casado-Lumbreras, C., & Cabezas-Isla, F. (2010). Shaping human capital in software development teams:
the case of mentoring enabled by semantics. IET Softw, 4(6), 445–452. ,
,
,
,
,
(
)
p g
the case of mentoring enabled by semantics. IET Softw, 4(6), 445–452. Soto-Acosta, P, Popa, S & Palacios-Marqués, D (2015). E-business, organizational innovation and firm performance
in manufacturing SMEs: an empirical study in Spain. Technological and Economic Development of Economy,
http://dx.doi.org/10.3846/20294913.2015.1074126 Taylor, M. Z., & Wilson, S. (2012). Does culture still matter? The effects of individualism on national innovation rates. J Bus Venturing, 27(2), 234–247. Tham, J. (2010). Corporate social innovation. Soc Space, 1, 48–56. Trequattrini, R., Lombardi, R., & Battista, M. (2015). Network analysis and football team performance: a first application. Team Perfor Manag, 21(1/2), 85–110. Webb, T (2007). Strategy & management: Unilever’s CEO: social innovation and sustainability the only game in town. Ethical Corporation, 5. Zahra, S., Gedajlovic, E., Neubaum, D., & Schulman, J. (2009). A typology of social entrepreneurs: motives, search
processes and ethical challenges. J Bus Venturing, 24(5), 519–532. Canestrino et al. Journal of Innovation and Entrepreneurship (2015) 4:12 References Mexico: Instituto Tecnologico y de Estudios Superiores de Monterrey and Instituto de Empresa. Kaasa, A., & Vadi, M. (2010). How does culture contribute to innovation? Evidence from European countries. Econ Innov
N Technol, 19(7), 583–604. Kanter, R. M. (1999). From spare change to real change: the social sector as beta site for business innovation. Harv Bus
Rev, 77, 122–133. ,
,
Kerlin, J. A. (Ed.). (2009). Social enterprise: a global comparison. New England: University Press of New England. erlin, J. A. (Ed.). (2009). Social enterprise: a global comparison. New Engl Krull, P. (2008). The emerging field of social innovation through corporate social responsibility—four tendencies, 4th EABIS
conference. UK: Cranfield University. Krull, P. (2008). The emerging field of soci
conference. UK: Cranfield University. Maggioni, V., & Del Giudice, M. (2011). Relazioni sistemiche tra imprenditorialità interna e gemmazione d’impresa: una
ricerca empirica Sulla natura cognitiva Delle nuove imprese. Sinergie, 71, 171–197. Mair, J., & Martí, I. (2006). Social entrepreneurship research: a source of explanation, prediction, and delight. J World Bus,
41(1), 36–44. Matten, D., & Moon, J. (2008). “Implicit” and “explicit” CSR: a conceptual framework for a comparative understanding of
corporate social responsibility. Acad Manage Rev, 33(2), 404–424. Meroño-Cerdan, A. L., & Soto-Acosta, P. (2005). Examining e-business impact on firm performance through website
analysis. Int J Electron Bus, 3(6), 583–598. Moores, T. T. (2003). The effect of national culture and economic wealth on global software piracy rates. Commun ACM,
46(9), 207–215. Mulgan, G. (2006). The process of social innovation, technology, governance, globalization. MIT Press, 1(2), 145–162. Nonaka, I., & Takeuchi, H. (1995). The knowledge-creating company. Oxford: Oxford University Press. Palacios-Marqués, D., Soto-Acosta, P., & Merigó, J. M. (2015). Analyzing the effects of technologica
competition factors on web knowledge exchange in SMEs. Telematics Inform, 32(1), 23–32. Palacios Marqués, D., Soto Acosta, P., & Merigó, J. M. (2015). Analyzing the effects of technologi
competition factors on web knowledge exchange in SMEs. Telematics Inform, 32(1), 23–32. competition factors on web knowledge exchange in SMEs. Telematics Inform, 32(1), 23–32. Parjanen, S. (2012). Experiencing creativity in the organization: from individual creativity to collective creativity. Interdisciplinary J Inf Knowl Manag, 7, 109–128. Petrakis, P. E., & Pantelis, C. K. (2014). Medium term effects of culture, transactions and institutions on opportunity
entrepreneurship. J Innov Entrep, 3(11), 1–22. Razavi, S. M., Asadi, M., Esfandabadi, H. M., & Ekbatani, H. (2014). References Submit your manuscript to a
journal and benefi t from:
7 Convenient online submission
7 Rigorous peer review
7 Immediate publication on acceptance
7 Open access: articles freely available online
7 High visibility within the fi eld
7 Retaining the copyright to your article
Submit your next manuscript at 7 springeropen.com Submit your manuscript to a
journal and benefi t from:
7 Convenient online submission
7 Rigorous peer review
7 Immediate publication on acceptance
7 Open access: articles freely available online
7 High visibility within the fi eld
7 Retaining the copyright to your article
Submit your next manuscript at 7 springeropen.com Submit your manuscript to a
journal and benefi t from:
7 Convenient online submission
7 Rigorous peer review
7 Immediate publication on acceptance
7 Open access: articles freely available online
7 High visibility within the fi eld
7 Retaining the copyright to your article
Submit your next manuscript at 7 springeropen.com
|
https://openalex.org/W4240433802
|
https://europepmc.org/articles/pmc5492161?pdf=render
|
English
| null |
The Impact of Aerobic Exercise on Fronto-Parietal Network Connectivity and Its Relation to Mobility: An Exploratory Analysis of a 6-Month Randomized Controlled Trial
|
Frontiers in human neuroscience
| 2,017
|
cc-by
| 11,332
|
ORIGINAL RESEARCH
published: 30 June 2017
doi: 10.3389/fnhum.2017.00344 Chun L. Hsu1,2,3,4, John R. Best1,2,3,4, Shirley Wang1,2,3,4, Michelle W. Voss5,6,
Robin G. Y. Hsiung7, Michelle Munkacsy1,2,3,4, Winnie Cheung1,2,3,4, Todd C. Handy8 and
Teresa Liu-Ambrose1,2,3,4* 1 Aging, Mobility, and Cognitive Neuroscience Lab, University of British Columbia, Vancouver, BC, Canada, 2 Department of
Physical Therapy, University of British Columbia, Vancouver, BC, Canada, 3 Djavad Mowafaghian Center for Brain Health,
University of British Columbia, Vancouver, BC, Canada, 4 Center for Hip Health and Mobility, Vancouver, BC, Canada,
5 Health, Brain, and Cognition Lab, University of Iowa, Iowa City, IA, United States, 6 Department of Psychology, University of
Iowa, Iowa City, IA, United States, 7 Department of Medicine, University of British Columbia, Vancouver, BC, Canada,
8 Department of Psychology, University of British Columbia, Vancouver, BC, Canada Keywords:
aging,
impaired
mobility,
vascular
cognitive
impairment,
fronto-parietal
network,
functional
connectivity, fMRI Edited by: Impaired mobility is a major concern for older adults and has significant consequences. While the widely accepted belief is that improved physical function underlies the
effectiveness of targeted exercise training in improving mobility and reducing falls, recent
evidence suggests cognitive and neural benefits gained through exercise may also play
an important role in promoting mobility. However, the underlying neural mechanisms of
this relationship are currently unclear. Thus, we hypothesize that 6 months of progressive
aerobic exercise training would alter frontoparietal network (FPN) connectivity during
a motor task among older adults with mild subcortical ischemic vascular cognitive
impairment (SIVCI)—and exercise-induced changes in FPN connectivity would correlate
with changes in mobility. We focused on the FPN as it is involved in top-down attentional
control as well as motor planning and motor execution. Participants were randomized
either to usual-care (CON), which included monthly educational materials about VCI
and healthy diet; or thrice-weekly aerobic training (AT), which was walking outdoors
with progressive intensity. Functional magnetic resonance imaging was acquired at
baseline and trial completion, where the participants were instructed to perform bilateral
finger tapping task. At trial completion, compared with AT, CON showed significantly
increased FPN connectivity strength during right finger tapping (p < 0.05). Across
the participants, reduced FPN connectivity was associated with greater cardiovascular
capacity (p = 0.05). In the AT group, reduced FPN connectivity was significantly
associated with improved mobility performance, as measured by the Timed-Up-and-Go
test (r = 0.67, p = 0.02). These results suggest progressive AT may improve mobility in
older adults with SIVCI via maintaining intra-network connectivity of the FPN. Impaired mobility is a major concern for older adults and has significant consequences. While the widely accepted belief is that improved physical function underlies the
effectiveness of targeted exercise training in improving mobility and reducing falls, recent
evidence suggests cognitive and neural benefits gained through exercise may also play
an important role in promoting mobility. However, the underlying neural mechanisms of
this relationship are currently unclear. Thus, we hypothesize that 6 months of progressive
aerobic exercise training would alter frontoparietal network (FPN) connectivity during
a motor task among older adults with mild subcortical ischemic vascular cognitive
impairment (SIVCI)—and exercise-induced changes in FPN connectivity would correlate
with changes in mobility. We focused on the FPN as it is involved in top-down attentional
control as well as motor planning and motor execution. Edited by: Participants were randomized
either to usual-care (CON), which included monthly educational materials about VCI
and healthy diet; or thrice-weekly aerobic training (AT), which was walking outdoors
with progressive intensity. Functional magnetic resonance imaging was acquired at
baseline and trial completion, where the participants were instructed to perform bilateral
finger tapping task. At trial completion, compared with AT, CON showed significantly
increased FPN connectivity strength during right finger tapping (p < 0.05). Across
the participants, reduced FPN connectivity was associated with greater cardiovascular
capacity (p = 0.05). In the AT group, reduced FPN connectivity was significantly
associated with improved mobility performance, as measured by the Timed-Up-and-Go
test (r = 0.67, p = 0.02). These results suggest progressive AT may improve mobility in
older adults with SIVCI via maintaining intra-network connectivity of the FPN. Edited by:
Srikantan S. Nagarajan,
University of California,
San Francisco, United States Reviewed by:
C. J. Boraxbekk,
Umeå University, Sweden
Gary Abrams,
University of California,
San Francisco, United States
Karl Meisel,
University of California,
San Francisco, United States *Correspondence:
Teresa Liu-Ambrose
teresa.ambrose@ubc.ca *Correspondence:
Teresa Liu-Ambrose
teresa.ambrose@ubc.ca Received: 14 November 2016
Accepted: 14 June 2017
Published: 30 June 2017 The Impact of Aerobic Exercise on
Fronto-Parietal Network Connectivity
and Its Relation to Mobility: An
Exploratory Analysis of a 6-Month
Randomized Controlled Trial Chun L. Hsu1,2,3,4, John R. Best1,2,3,4, Shirley Wang1,2,3,4, Michelle W. Voss5,6,
Robin G. Y. Hsiung7, Michelle Munkacsy1,2,3,4, Winnie Cheung1,2,3,4, Todd C. Handy8 and
Teresa Liu-Ambrose1,2,3,4* INTRODUCTION underlying white matter lesions (WMLs) or lacunar infarcts
(Roman et al., 2002; Vermeer et al., 2007). VCI is the second
most common cause of dementia after Alzheimer’s disease (AD)
(Desmond et al., 1999; Erkinjuntti et al., 1999; Pantoni et al.,
1999; Rockwood et al., 2000). The clinical consequences of
covert ischemic strokes are substantial (Roman et al., 2002;
Vermeer et al., 2007). These WMLs and lacunar infarcts typically
manifest in brain regions such as caudate, pallidum, thalamus,
frontal and prefrontal white-matter (Roman et al., 2002). As
a result, they may disrupt the integrity of functional neural
networks and negatively impact cognitive function, particulary
executive functions, and mobility (Kuo and Lipsitz, 2004;
O’Brien, 2006). Impaired mobility is a major concern for older adults and is
associated with increased risk for disability, institutionalization,
and death (Rosano et al., 2008). The prevalence of impaired
mobility is 14% at age 75 years and involves half of the population
over 84 years (Odenheimer et al., 1994). Falls are a significant
consequence of impaired mobility. Impaired mobility is a major concern for older adults and is
associated with increased risk for disability, institutionalization,
and death (Rosano et al., 2008). The prevalence of impaired
mobility is 14% at age 75 years and involves half of the population
over 84 years (Odenheimer et al., 1994). Falls are a significant
consequence of impaired mobility. Current evidence supports the recommendation of targeted
exercise training to improve mobility, prevent major mobility
disability, and reduce the risk of future falls (Campbell et al.,
1999; Pahor et al., 2014). The widely accepted view is that
improved physical function, such as improved balance and
increased muscle strength, primarily underlies the effectiveness
of the exercise in improving mobility and reducing falls (Layne
et al., 2017). However, in a meta-analysis of four randomized
trials of exercise, falls were significantly reduced by 35% while
postural sway significantly improved by only 9% and there was no
significant improvement in knee extension strength (Robertson
et al., 2002). Moreover, in a proof-of-concept randomized
controlled trial, we demonstrated that a home-based exercise
signficantly reduced falls by 47% in older adults – in the
absence of significant improvement in physical function (i.e.,
balance and muscle strength) (Liu-Ambrose et al., 2008). Rather,
significant improvement in executive functions were observed in
the exercise group as compared with the usual care (i.e., control)
group. INTRODUCTION Among the relevant neural networks, most notably, is the
frontoparietal network (FPN). The FPN is involved in top-
down attentional control and allocation of available neural
resources that contribute to executive processes, such as response
anticipation and conflict processing (Fogassi and Luppino, 2005;
Seeley et al., 2007; Sridharan et al., 2008), as well as motor
planning and motor execution (Wise et al., 1997; Wu et al.,
2009; Wymbs et al., 2012). Of particular relevance, previous
studies have shown that key regions within the FPN were actively
recruited during actual as well as imagined completion of the
walking while talking (WWT) test (Holtzer et al., 2011; Blumen
et al., 2014). Specifically, neural activity within these FPN regions
were positively associated with both task difficulty and cognitive
performance of the WWT test (Holtzer et al., 2011; Blumen
et al., 2014). Although previous studies have linked aspects of
mobility to FPN connectivity, its potential role understanding
exercise-induced effects on mobility is unkown. These data suggest that exercise may reduce falls in older
adults via several mechanisms, not just via improved physical
function. y
Thus, we propose FPN connectivity as one of the neural
mechanisms by which exercise promotes mobility in older adults
with mild SIVCI. Using functional magnetic resonance imaging
(fMRI) data from a 6-month single-blind randomized controlled
trial (clinicaltrials.gov Identifier: NCT01027858), we conducted
a planned secondary analysis to assess the impact of moderate-
intensity aerobic exercise training on functional connectivity of
FPN among older adults with mild SIVCI. We hypothesized that
aerobic exercise-induced increases in FPN connectivity would
correlate with improved mobility. The primary results from the
parent study have been published (Liu-Ambrose et al., 2016),
which provided preliminary evidence that 6 months of thrice-
weekly progressive aerobic training (AT) promotes cognitive
performance in community-dwelling adults with mild SIVCI,
relative to usual care plus education. We previously proposed that cognitive and neural plasticity
may be an important, yet under-appreciated mechanism by
which exercise promotes mobility and reduce falls (Liu-Ambrose
et al., 2013). INTRODUCTION This hypothesis stems from the growing evidence
that suggest: (1) cognitive impairment and impaired mobility are
associated (Atkinson et al., 2007; Buracchio et al., 2010; Montero-
Odasso et al., 2012); (2) reduced executive function, is associated
with impaired mobility and increased falls risk (Delbaere et al.,
2012; Hsu et al., 2012); (3) aberrant neural network functional
connectivity is associated with impaired mobility (Hsu et al.,
2014); and (4) targeted exercise training, particulary aerobic-
based, promotes cognitive and cortical plasticity, including
executive function and its neural correlates, in older adults
(Erickson and Kramer, 2009; Voss et al., 2010b). Despite the growing recognition that targeted exercise training
may promote mobility outcomes in older adults via central
mechanisms (Liu-Ambrose et al., 2013), few intervention studies
of exercise to date have provided direct evidence for this
theory (Bolandzadeh et al., 2015). A better understanding of the
neural mechanisms underlying exercise-induced improvements
in mobility may facilitate the development and refinement
of preventative/intervention strategies, as well as identify the
populations for whom these effects apply. Citation: Hsu CL, Best JR, Wang S,
Voss MW, Hsiung RGY,
Munkacsy M, Cheung W, Handy TC
and Liu-Ambrose T (2017)
The Impact of Aerobic Exercise on
Fronto-Parietal Network Connectivity
and Its Relation to Mobility: An
Exploratory Analysis of a 6-Month
Randomized Controlled Trial. Front. Hum. Neurosci. 11:344. doi: 10.3389/fnhum.2017.00344 Hsu CL, Best JR, Wang S,
Voss MW, Hsiung RGY,
Munkacsy M, Cheung W, Handy TC
and Liu-Ambrose T (2017)
The Impact of Aerobic Exercise on
Fronto-Parietal Network Connectivity
and Its Relation to Mobility: An
Exploratory Analysis of a 6-Month
Randomized Controlled Trial. Front. Hum. Neurosci. 11:344. doi: 10.3389/fnhum.2017.00344 June 2017 | Volume 11 | Article 344 Frontiers in Human Neuroscience | www.frontiersin.org 1 Understanding the Neural Underpinnings of Exercise Hsu et al. Frontiers in Human Neuroscience | www.frontiersin.org Participants p
As the current study was a secondary analysis, we sought to
recruit as many eligible and consenting individuals from the
parent study as possible. To briefly describe the recruitment
process, we recruited from the University of British Columbia
Hospital Clinic for Alzheimer’s Disease and Related Disorders,
the Vancouver General Hospital Stroke Prevention Clinic, and
specialized geriatric clinics in Metro Vancouver, BC. Recruitment
occurred
between
December
2009
and
April
2014
with
randomization occurring on an ongoing basis. Study participants
were clinically diagnosed with mild SIVCI as determined by
the presence of cognitive syndrome and small vessel ischaemic
disease (Erkinjuntti et al., 2000). Small vessel ischemic disease
was defined as evidence of relevant cerebrovascular disease by
brain computed tomography or MRI defined as the presence of
both: (1) Periventricular and deep WMLs; (2) Absence of cortical
and/or cortico-subcortical non-lacunar territorial infarcts and
watershed infarcts, hemorrhages indicating large vessel disease,
signs of normal pressure hydrocephalus, or other specific causes
of WMLs (i.e., multiple sclerosis, leukodystrophies, sarcoidosis,
brain irradiation). In addition to the neuroimaging evidence, the
presence or a history of neurological signs such as Babinski sign,
sensory deficit, gait disorder, or extrapyramidal signs consistent
with sub-cortical brain lesion(s) was required and confirmed The Consolidated Standards of Reporting Trial flowchart
shows the number and distribution of participants included in
this secondary analysis (Figure 1). Of the 38 participants (54%
of parent sample) that completed baseline MRI scanning, 7 (18%
of the sample) dropped out from the study and 10 (26% of the
sample) failed to correctly perform the motor finger tapping
task (e.g., finger tapped during resting blocks). Consequently, 21
participants who completed MRI at baseline and trial completion
were included in this secondary analysis (30% of parent sample). Ethical approval was provided by the University of British
Columbia’s Clinical Research Ethics Board (H07-01160). All
participants provided written informed consent. Study Design This is a secondary analysis of neuroimaging data acquired
from a 6-month proof-of-concept RCT (NCT01027858) of
aerobic exercise in older adults with mild SIVCI (Liu-Ambrose
et al., 2010, 2016). Trained study assessors were blinded to
group allocation of participants. Functional MRI (fMRI) data
were acquired at baseline prior to randomization and at trial
completion (i.e., 6 months) in a subset of eligible participants. Older adults with subcortical ischemic vascular cognitive
impairment (SIVCI), the most common form of vascular
cognitive impairment (VCI) (Bowler, 2005), are at particular
risk for both impaired mobility and dementia secondary to June 2017 | Volume 11 | Article 344 Frontiers in Human Neuroscience | www.frontiersin.org 2 Understanding the Neural Underpinnings of Exercise Hsu et al. by study physicians (G-YRH and PL). Cognitive syndrome was
defined as a baseline Montreal Cognitive Assessment (MoCA)
score less than 26/30. However, participants were free of frank
dementia (i.e., clinically diagnosis of dementia) as determined by
a Mini-Mental State Examination (MMSE) score ≥20 and the
absence of diagnosed dementia. Progressive cognitive decline was
confirmed through medical records or caregiver/family member
interviews. Descriptive Variables or usual care (CON). A research team member not involved
with the study held this sequence at a remote location. After
the completion of consent and baseline testing, the research
coordinator contacted the team member holding the list to
determine the next allocation. At baseline, participants underwent a clinical assessment with
study physicians (GYRH and PL) to confirm current health
status and study eligibility. Age in years and education level
were assessed by self-report. Standing height was measured as
stretch stature to the 0.1 cm per standard protocol. Weight was
measured twice to the 0.1 kg on a calibrated digital scale. Waist-
to-hip ratio was determined by measuring the widest part of
the hip circumference and the waist just above the navel in
centimeters. The Functional Comorbidity Index (Groll et al.,
2005) assessed the number of comorbid conditions related to
physical functioning. Usual Care Participants in the CON group received usual care, in which
they were provided with monthly educational materials about
VCI and healthy diet. However, no specific information regarding
physical activity was provided. In addition, research staffphoned
the CON participants on a monthly basis to maintain contact and
to acquire research data. Functional MRI Acquisition q
All MRI was conducted at the University of British Columbia
(UBC) MRI Research Center located at the UBC Hospital
on a 3.0 Tesla Intera Achieva MRI Scanner (Philips Medical
Systems, Markham, ON, Canada) using an 8-channel SENSE
neurovascular coil. The fMRI consisted of two successive runs
with 165 dynamic images of 36 slices (3 mm thick) with the
following parameters: repetition time (TR) of 2000 ms, echo
time (TE) of 30 ms, flip angle (FA) of 90 degrees, field of view
(FoV) of 240 mm, acquisition matrix 80 × 80, voxel size of
3 mm × 3 mm × 3 mm. High resolution anatomical MRI T1
images were acquired using the following parameters: 170 slices
(1 mm thick), TR of 7.7 ms, TE of 3.6 ms, FA of 8 degrees, FoV of
256 mm, acquisition matrix of 256 × 200. The AT group was also given a pedometer to serve as both an
incentive and monitoring tool. Participants recorded the number
of steps each day taken outside the AT classes on standard logs
provided by the research team. During each scanning session, the study participants were
asked to perform a finger tapping motor task that had been
previously administered and described (Hsu et al., 2014). Briefly,
the task consisted of three conditions: right finger tapping, rest,
and left finger tapping. The specific instructions given required
the participants to finger tap in a particular sequence regardless
of condition: start with the index finger and progress toward the
little (pinky) finger continuously until a different condition is
presented. For the rest condition, participants were asked to rest
with their eyes open. The exact order of motor task blocks was
not disclosed to the participants and was counter balanced over
two runs as follow: Randomization The randomization sequence was generated using the web
application www.randomization.com with a ratio of 1:1 to AT FIGURE 1 | Overview of the flow of study participants through the 6-month study. FIGURE 1 | Overview of the flow of study participants through the 6-month study. June 2017 | Volume 11 | Article 344 3 Understanding the Neural Underpinnings of Exercise Hsu et al. Aerobic Training and Compliance For the AT group, AT consisted of supervised thrice-weekly
60-min classes of walking for the 6-month intervention period. All AT group classes were led by instructors certified to
instruct seniors and were delivered in a group setting. Each
60-min class included a 10-min warm-up, 40-min of walking,
and a 10-min cool down. Both the warm-up and cool-down
included passive and active stretches, as well as range of motion
exercise. Global cognition was assessed using the MMSE (Cockrell
and Folstein, 1988) and the MoCA. The MMSE and MoCA are
30-point tests that encompass several cognitive domains. The
MoCA has been found to have good internal consistency and
test–retest reliability and was able to correctly identify 90% of a
large sample of individuals with mild cognitive impairment from
two different clinics with a cut-offscores of ≤26/30 (Nasreddine
et al., 2005). Walking occurred outdoors and followed predetermined
routes around local areas. The intensity of the AT program was
monitored and progressed using three approaches: (1) heart rate
monitoring with an initial intensity of 40% of age specific target
heart rate (i.e., heart rate reserve; HRR). HRR was calculated by
subtracting resting heart rate from maximum heart rate [using
the formula: 206.9 – 0.67 × Age (Gellish et al., 2007)] and
recalculation each month. Participants progressed over the first
12 weeks to the range of 60–70% of HRR, after which this
was sustained for the remainder of the intervention period;
(2) subjective monitoring using the Borg’s Rating of Perceived
Exertion (RPE) (Borg, 1982) with a target RPE of 14 to 15;
and (3) the “talk” test (Persinger et al., 2004), starting at a
walking pace allowing comfortable conversation and progressing
to a walking pace where conversation was difficult. Individual
training logs (i.e., target heart rate, heart rate achieved, and rate of
perceived exertion) were maintained throughout the intervention
period. Frontiers in Human Neuroscience | www.frontiersin.org Mobility, Cardiovascular Capacity, and
Physical Activity From each ROI, preprocessed time-series data were extracted
with 14 mm spherical regions of interest drawn around their
respective MNI coordinates in standard space. The different
conditions (i.e., left, right, and rest) within each block of
the motor task were extracted and compiled together. To
concatenate the time-series data, the stimulus onset time for
each task condition was acquired from the task program. Each volume of the data was then sorted according to their
respective condition. Once the data were properly categorized,
the task-specific volumes (e.g., all the “left” volumes) were
merged using a script provided in the FSL program. The first
three volumes of any condition were discarded to account for
delay of the hemodynamic response. Evidence in the literature
has demonstrated that functional connectivity derived from
temporally spliced/merged resting-state data from blocked fMRI
design is not significantly different from connectivity derived
from continuous data (Fair et al., 2007). Recent study using
motor task fMRI also showed that quantifying functional
connectivity via similar seed-based approach using concatenated
data is comparable to results from continuous data (Zhu et al.,
2017). Mobility was assessed with the Timed-Up-and-Go test (TUG)
and the Short Physical Performance Battery (SPPB). The TUG
required participants to rise from a standard chair, walk a
distance of three meters, turn, walk back to the chair and sit
down (Shumway-Cook et al., 2000). We recorded the time (s) to
complete the TUG, based on the average of two separate trials. p
g
p
For the SPPB, participants were assessed on performances
of standing balance, walking, and sit-to-stand. Each component
is rated out of four points, for a maximum of 12 points; a
score < 9/12 predicts subsequent disability (Guralnik et al., 1995). Participant’s cardiovascular capacity was assessed using the
6-Minute Walk Test (Enright, 2003). The total distance walked
(meters) within the span of 6 min was recorded. For the SPPB, participants were assessed on performances
of standing balance, walking, and sit-to-stand. Each component
is rated out of four points, for a maximum of 12 points; a
score < 9/12 predicts subsequent disability (Guralnik et al., 1995). Participant’s cardiovascular capacity was assessed using the
6-Minute Walk Test (Enright, 2003). The total distance walked
(meters) within the span of 6 min was recorded. Monthly total physical activity level was determined by
the Physical Activities Scale for the Elderly (PASE) self-report
questionnaire (Washburn et al., 1999). Functional Connectivity Analysis Previous studies guided our choice of seeds in the whole brain
analysis of the FPN (Voss et al., 2010b). The FPN included
the inferior parietal sulcus (IPS), ventral visual cortex (VV),
supramarginal gyrus (SMG), superior lateral occipital cortex
(SLOC), frontal eye field (FEF), as well as overlapping areas
in the temporal-parietal junction. The respective MNI space
coordinates for each region of interest (ROI) are provided in
Table 1. Adverse Effects Briefly, WMLs were
identified and digitally marked (i.e., placing seed points) by
a radiologist on T2 and PD weighted images. Marked WMLs
were automatically segmented by a customized Parzen windows
classifier that estimated the intensity distribution of the lesions –
which also included heuristics that optimized the accuracy of the
estimated distributions (Parzen, 1962; McAusland et al., 2010;
Bolandzadeh et al., 2015). WML segmentation was reviewed by
a trained technician to ensure accuracy. Noise
generated
from
both
physiological
and
non-
physiological sources were removed through regression of
the cerebral-spinal fluid (CSF) signal, white matter signal, and
global brain signal. Global signal regression had been reported
as both valid and useful step in functional connectivity analyses
(Fox et al., 2009) that may improve specificity (Murphy and Fox,
2016). Adverse Effects All participants were instructed to report any adverse effects due
to the AT exercises to our research coordinator, such as falls or
musculoskeletal pain persisting longer than 48 h. Participants
were also questioned about the presence of any adverse effects,
such as musculoskeletal pain or discomfort, at each exercise
session. All instructors also monitored participants for symptoms
of angina and shortness of breath during the exercise classes. External experts from our safety monitoring committee reviewed
all adverse events reported on a monthly basis. Run A: Rest, Left Tap, Rest, Right Tap, Rest, Right Tap, Rest,
Left Tap, Rest
Run B: Rest, Right Tap, Rest, Left Tap, Rest, Left Tap, Rest,
Right Tap, Rest Total WML volume (in mm3) at baseline was quantified
with structural MRI data acquired on the same MRI scanner
(3T Achieva, Philips Medical Systems, Markham, ON, Canada)
at the UBC MRI Research Centre. A T2-weighted scan and June 2017 | Volume 11 | Article 344 Frontiers in Human Neuroscience | www.frontiersin.org 4 Understanding the Neural Underpinnings of Exercise Hsu et al. a proton-density-weighted (PD-weighted) scan were acquired
for each subject. For the T2-weighted images, the repetition
time (TR) was 5,431 ms and the echo time (TE) was 90 ms,
and for the PD-weighted images, the TR was 2,000 ms,
and the TE was 8 ms. T2- and PD-weighted scans had
dimensions of 256 × 256 × 60 voxels and a voxel size of
0.937 mm × 0.937 mm × 3.000 mm. Briefly, WMLs were
identified and digitally marked (i.e., placing seed points) by
a radiologist on T2 and PD weighted images. Marked WMLs
were automatically segmented by a customized Parzen windows
classifier that estimated the intensity distribution of the lesions –
which also included heuristics that optimized the accuracy of the
estimated distributions (Parzen, 1962; McAusland et al., 2010;
Bolandzadeh et al., 2015). WML segmentation was reviewed by
a trained technician to ensure accuracy. a proton-density-weighted (PD-weighted) scan were acquired
for each subject. For the T2-weighted images, the repetition
time (TR) was 5,431 ms and the echo time (TE) was 90 ms,
and for the PD-weighted images, the TR was 2,000 ms,
and the TE was 8 ms. T2- and PD-weighted scans had
dimensions of 256 × 256 × 60 voxels and a voxel size of
0.937 mm × 0.937 mm × 3.000 mm. Frontiers in Human Neuroscience | www.frontiersin.org Statistical Analyses Statistical analysis was conducted using the IBM SPSS Statistic 23
for Windows (SPSS Inc., Chicago, IL, United States). Statistical
significance was set at p ≤0.05 for all analyses. Change
in network connectivity strength was computed in SPSS as
6-month FPN connectivity minus baseline FPN connectivity. Linear mixed models with random intercepts and time-varying
outcome measures were constructed to statistically test for
significant between-group differences in change in network
connectivity while adjusting for baseline total WML and age. A group by time interaction indicated group differences in
changes in FPN connectivity from baseline to post-intervention. Similar analyses were conducted to determine whether there
were group differences in changes in TUG, SPPB, 6MWT, and
PASE scores. The primary analyses included the 21 participants
with valid baseline and post-intervention fMRI data. Secondary
analyses followed the intention-to-treat principle by including
nine additional individuals with valid baseline fMRI but were
lost to follow-up; maximum likelihood estimation allowed for
these individuals to inform the treatment effects, despite having
missing follow-up data and to determine whether loss to follow-
up might bias the treatment effects estimated with only treatment
completers. MMSE, Mini-Mental Status Examination; MoCA, Montreal Cognitive Assessment;
FCI, Functional Comorbidity Index. All measures were not statistically significant at
p < 0.05. TABLE 3 | AT group pedometer information over 6-month intervention period. TABLE 3 | AT group pedometer information over 6 month intervention period. Average pedometer count
Mean (SD)
Baseline
7002.1 (5496.4)
Month 1
9009.2 (6337.9)
Month 2
9786.3 (6944.2)
Month 3
10713.8 (7209.0)
Month 4
10922.1 (7645.7)
Month 5
11484.8 (8074.6)
Month 6
11552.7 (7411.9) age of all participants included in this secondary analysis was
71.1 years (SD = 8.7 years), which is not significantly different
from the mean age of the parent cohort at 74.3 years (SD = 8.3
years). Compared to the nine individuals with valid baseline data
only, the study sample did not differ on baseline FPN connectivity
(all p > 0.19) but did have higher average baseline MoCA scores
(23.2 versus 20.4; p = 0.02). Neither of the mobility measures
nor self-reported physical activity differed significantly between
groups across the 6 months. However, we observed a trend-
level group difference in the change in 6-Minute Walk Test
performance (p = 0.08; Table 4), in which the AT group showed
greater improvement (48.6 m) compared with the CON group
(−0.3 m). Data Analysis
Functional MRI Preprocessing Image preprocessing was carried out using tools from FSL
(FMRIB’s Software Library) [78], MATLAB (Matrix Laboratory),
and toolboxes from SPM (Statistical Parametric Mapping). Excess
unwanted structures (i.e., bones, skull, etc.) in high resolution
T1 images were removed via Brain Extraction Tool (BET);
rigid body motion correction was completed using MCFLIRT
(absolute and relative mean displacement were subsequently
extracted and included in the statistical analysis as covariates);
spatial smoothing was carried out using Gaussian kernel of Full-
Width-Half-Maximum (FWHM) 6.0 mm; temporal filtering was
applied with high pass frequency cut-offof 120 s. In addition,
a low pass temporal filtering was also included to ensure the
fMRI signal fluctuated between 0.008 < f < 0.080 Hz, the ideal
bandwidth to examine functional connectivity. Furthermore, the
application of a low pass filter eliminated high frequency signals
that could be confounds. Participants’ low-resolution functional
data were registered to personal high resolution T1 anatomical
images, which were subsequently registered to standardized 152
T1 Montreal Neurological Institute (MNI) space. Region of interest time-series data were subsequently cross-
correlated with every voxel within the brain to establish
functional connectivity maps of their associated neural networks, TABLE 1 | Frontoparietal network regions of interest and relative MNI coordinates. FPN
ROI
X
Y
Z
RIPS
25
−62
53
RVV
36
−62
0
LVV
−44
−60
−6
RSMG
32
−38
38
RSLOC
26
−64
54
LSLOC
−26
−60
52
RFEF
28
−4
58
LFEF
−26
−8
54
RIPS, right inferior parietal sulcus; RVV, right ventral visual; LVV, left ventral visual;
RSMG, right supramarginal gyrus; RSLOC, right lateral occipital cortex; LSLOC, left
lateral occipital cortex; RFEF, right frontal eye field; LFEF, left frontal eye field. TABLE 1 | Frontoparietal network regions of interest and relative MNI coordinates. Frontiers in Human Neuroscience | www.frontiersin.org Frontiers in Human Neuroscience | www.frontiersin.org June 2017 | Volume 11 | Article 344 5 Understanding the Neural Underpinnings of Exercise Hsu et al. TABLE 2 | Participant characteristics at baseline (N = 21). Statistical Analyses Bivariate correlation analyses were performed to determine
whether
any
significant
changes
in
intra-network
FPN
connectivity (during rest, left tap, and right tap) in the AT
group (n = 12) correlated with change in mobility, as measured
by TUG and SPPB, or change in 6MWT across the 6-month
study duration. AT Compliance and Adverse Effects Participants and Treatment Fidelity
Among the 70 randomized individuals in the parent study, we
observed a significant effect of AT on 6-min walk performance,
a well-established tool that accurately evaluates cardiovascular
fitness (Cataneo et al., 2010) (B = 30.34, p = 0.02), indicating that
AT had a positive effect on cardiovascular capacity (Liu-Ambrose
et al., 2016). Twenty-one participants who completed fMRI scans
at both baseline and 6 months were included in the primary
analysis (Figure 1). Study demographics are reported in Table 2,
pedometer information over the intervention period is reported
in Table 3, mobility and cardiovascular capacity measures are
reported in Table 4; these measures do not differ between groups
at baseline nor differ significantly from the 70 eligible participants
enrolled in the parent study (Liu-Ambrose et al., 2016). The mean The average compliance observed in the AT group was 76% for
the walking classes and 65% for the nutrition education classes;
whereas the average compliance observed in the CON group was
74%. Two study-related adverse events were reported in the AT
group and one in the CON group. All three were non-syncopal
falls. One of the falls in the AT group resulted in a broken
tooth and required assessment in the Emergency Department; the
remaining two did not result in injury. Data Analysis
Functional MRI Preprocessing CON group, n = 9
AT Group, n = 12
Variables
Mean (SD) or n
Mean (SD) or n
Age (year)
69.9 (9.2)
72.0 (8.6)
Height (cm)
165.4 (11.4)
169.0 (15.9)
Weight (kg)
73.6 (13.1)
73.6 (14.8)
Sex (M/F)
5/4
8/4
MMSE (30 points max)
27.2 (1.9)
26.4 (2.8)
MOCA (30 points max)
24.2 (2.3)
22.5 (2.0)
FCI
3.0 (1.9)
3.1 (1.7)
White matter lesion (mm3)
1389.5 (2023.4)
3492.8 (3882.1)
Relative head motion (mm)
0.17 (0.09)
0.14 (0.07)
MMSE, Mini-Mental Status Examination; MoCA, Montreal Cognitive Assessment;
FCI, Functional Comorbidity Index. All measures were not statistically significant at
p < 0.05. TABLE 2 | Participant characteristics at baseline (N = 21). in which pairwise correlation between time-series extracted
from ROI listed above was calculated. Individual-level within-
subject results were generated via ordinary least-squares (OLS)
regression using FSL’s flameo (Beckmann et al., 2003) in
FSL by congregating the voxel-wise functional connectivity
maps from each condition. Similarly, for group results, a
mixed-level OLS analysis was conducted. The statistical map
thresholding was set at Z = 2.33, with cluster correction of
p < 0.05. Correlation Results Correlation Results group differences in the mean network connectivity strength
at baseline, regardless of task conditions (Table 5). At trial
completion, compared with AT, CON exhibited significantly
greater intra-network coupling of the FPN during right finger
tapping (p < 0.02) after adjusting for baseline WML and age. No AT effects were observed for FPN connectivity during left
finger tapping (p = 0.26) or during rest (p = 0.50). We conducted
a secondary, intention-to-treat analysis using all 30 participants
with usable baseline data, regardless of loss to follow-up, and
observed similar, though weaker, between-group differences in
FPN coupling during right finger tapping (p = 0.08). As with
the primary analyses, CON showed an increase in intra-network
coupling of the FPN (mean = 0.18, SE = 0.09), whereas
AT showed no significant change over time (mean = −0.04,
SE = 0.08). group differences in the mean network connectivity strength
at baseline, regardless of task conditions (Table 5). At trial
completion, compared with AT, CON exhibited significantly
greater intra-network coupling of the FPN during right finger
tapping (p < 0.02) after adjusting for baseline WML and age. No AT effects were observed for FPN connectivity during left
finger tapping (p = 0.26) or during rest (p = 0.50). We conducted
a secondary, intention-to-treat analysis using all 30 participants
with usable baseline data, regardless of loss to follow-up, and
observed similar, though weaker, between-group differences in
FPN coupling during right finger tapping (p = 0.08). As with
the primary analyses, CON showed an increase in intra-network
coupling of the FPN (mean = 0.18, SE = 0.09), whereas
AT showed no significant change over time (mean = −0.04,
SE = 0.08). Bivariate correlation across the study sample showed that the
change in FPN connectivity during right tapping was significantly
associated with change in 6-Minute Walk Test performance
(r = −0.43, p = 0.05; Table 6). Within the AT group (N = 12), the
change in FPN connectivity during right tapping was significantly
associated with change in TUG performance (r = 0.67, p = 0.02;
Table 6). Specifically, reduced FPN connectivity from baseline to
post-intervention correlated with improved TUG performance
over the same period of time (Figure 3). fMRI Results Baseline
Change from baseline to month 6∗(end of intervention)
Mean (SD)
Adjusted within-group change (SE)
Task condition
CON
AT
CON
AT
Between-group p-value
Right Tapping
0.25 (0.28)
0.34 (0.22)
0.25 (0.09)
−0.06 (0.07)
0.02
Rest
0.29 (0.24)
0.26 (0.24)
0.06 (0.09)
−0.02 (0.07)
0.50
Left Tapping
0.35 (0.24)
0.27 (0.20)
−0.05 (0.08)
0.08 (0.07)
0.26
∗Adjusted for baseline WML and age. TABLE 5 | Frontoparietal network connectivity during task (N = 21). fMRI Results Results from the seed-based functional connectivity analysis on
the FPN (Figure 2) showed there were no significant between Frontiers in Human Neuroscience | www.frontiersin.org Frontiers in Human Neuroscience | www.frontiersin.org June 2017 | Volume 11 | Article 344 6 Understanding the Neural Underpinnings of Exercise Hsu et al. TABLE 4 | Mobility and cardiovascular capacity measures (N = 21). Baseline
Change from baseline to month 6∗(end of intervention)
Mean (SD)
Adjusted within-group change (SE)
Variables
CON
AT
CON
AT
Between-group p-value
Timed Up and Go (s)
8.2 (1.3)
7.6 (1.8)
0.16 (0.46)
−0.06 (0.41)
0.73
Short Physical Performance Battery (max 12 points)
10.4 (0.9)
11.2 (1.3)
0.48 (0.34)
−0.23 (0.31)
0.15
Six-Minute Walk Test (meters)
531.2 (56.9)
555.6 (104.3)
−0.30 (19.29)
48.64 (16.57)
0.08
PASE Score
132.2 (61.3)
134.6 (94.0)
−6.05 (39.62)
−4.99 (34.05)
0.99
∗Adjusted for baseline WML and age; PASE, Physical Activities Scale for the Elderly. FIGURE 2 | Image of the fronto-parietal network. TABLE 5 | Frontoparietal network connectivity during task (N = 21). Baseline
Change from baseline to month 6∗(end of intervention)
Mean (SD)
Adjusted within-group change (SE)
Task condition
CON
AT
CON
AT
Between-group p-value
Right Tapping
0.25 (0.28)
0.34 (0.22)
0.25 (0.09)
−0.06 (0.07)
0.02
Rest
0.29 (0.24)
0.26 (0.24)
0.06 (0.09)
−0.02 (0.07)
0.50
Left Tapping
0.35 (0.24)
0.27 (0.20)
−0.05 (0.08)
0.08 (0.07)
0.26
∗Adjusted for baseline WML and age. group differences in the mean network connectivity strength
Correlation Results TABLE 4 | Mobility and cardiovascular capacity measures (N = 21). Baseline
Change from baseline to month 6∗(end of intervention)
Mean (SD)
Adjusted within-group change (SE)
Variables
CON
AT
CON
AT
Between-group p-value
Timed Up and Go (s)
8.2 (1.3)
7.6 (1.8)
0.16 (0.46)
−0.06 (0.41)
0.73
Short Physical Performance Battery (max 12 points)
10.4 (0.9)
11.2 (1.3)
0.48 (0.34)
−0.23 (0.31)
0.15
Six-Minute Walk Test (meters)
531.2 (56.9)
555.6 (104.3)
−0.30 (19.29)
48.64 (16.57)
0.08
PASE Score
132.2 (61.3)
134.6 (94.0)
−6.05 (39.62)
−4.99 (34.05)
0.99
∗Adjusted for baseline WML and age; PASE, Physical Activities Scale for the Elderly. TABLE 5 | Frontoparietal network connectivity during task (N = 21). DISCUSSION Contrary to our initial hypothesis, we found that a 6-
month AT intervention significantly alters FPN connectivity June 2017 | Volume 11 | Article 344 Frontiers in Human Neuroscience | www.frontiersin.org Frontiers in Human Neuroscience | www.frontiersin.org 7 Understanding the Neural Underpinnings of Exercise Hsu et al. TABLE 6 | Changes in mobility and changes in FPN connectivity correlation. 1Timed
1Short physical
1Six-minute
up and go
performance battery
walk test
Group (N = 21)
1FPN Connectivity
during right tapping
0.14
0.34
−0.43∗
AT group (n = 12)
1FPN Connectivity
during right tapping
0.67∗
0.14
−0.37
1 = 6-Months – Baseline. ∗p < 0.05. TABLE 6 | Changes in mobility and changes in FPN connectivity correlation. compared with age-matched healthy controls, individuals who
suffered an acute stroke showed significantly less left–right
posterior intraparietal sulcus connectivity while performing
a spatial-orientation task. Lower connectivity between the
left and right posterior intraparietal sulcus were significantly
associated with poorer task accuracy and slower task reaction
time. Moreover, current evidence suggests that SIVCI is generally
associated with less functional connectivity of neural networks. For example, among those with SIVCI, Yi et al. (2012) found
lower functional connectivity in the medial prefrontal cortex
and the middle temporal gyrus. Yu et al. (2015) demonstrated
that compared with healthy controls, individuals with SIVCI
had less network efficiency in the fronto-temporal and parietal
regions. Nonetheless, aberrant functional connectivity has been
repeatedly observed among those with SIVCI (Yi et al., 2012;
Ding et al., 2015; Yu et al., 2015; Zhou et al., 2016), and
the pattern (i.e., increased or decreased connectively) is not
consistent across studies. For instance, Ding et al. (2015) found
increased functional connectivity in the left middle temporal
lobe, right inferior temporal lobe, and left superior frontal
gyrus among patients with SIVCI as compared with healthy
controls. 1 = 6-Months – Baseline. ∗p < 0.05. during right finger tapping among older adults with mild
SIVCI. The observed effect of aerobic exercise on the FPN
during right tapping was significantly associated with improved
mobility and cardiovascular capacity. While these results are
preliminary, our data suggest aerobic exercise may promote
mobility among older adults with mild SIVCI via altering FPN
connectivity. y
Our findings are in contrast to previous findings that show
altered FPN connectivity is associated with aging (Andrews-
Hanna et al., 2007) and with cognitive deficits (He et al.,
2007; Geerligs et al., 2012; Poppe et al., 2015). DISCUSSION In addition, our data is limited
by the fact that only the connectivity during right hand tapping
was statistically significant while left hand was not. Differences in
social interactions experienced by the experimental groups may
present addition confounding factors to our data. Specifically,
active attention provided by trainers within the AT group
may potentially influence our findings. Lastly, the relationship
between connectivity and SIVCI status is equivocal with much
of the evidence generated from cross-sectional studies. Thus,
the inclusion of fMRI data from a healthy-aged matched cohort
might have facilitated interpretation of our results. Nevertheless,
we highlight the key strengths of our currents study design – a
randomized controlled trial – which are: (1) provides evidence
of causation; and (2) increased internal validity. Thus, our
study provides preliminary evidence to suggest that aerobic
exercise may impact functional connectivity in older adults Our observation that AT impacted FPN connectivity only
during right hand finger tapping concurs with the literature
that suggests the FPN connectivity is lateralized (Smith et al.,
2009; Pool et al., 2014; Gao et al., 2015). Specifically, using
independent component analysis, Smith et al. (2009) revealed
that among the neural networks identified, only the FPN exhibit
distinct left-right lateralized components. Also, Jancke et al. (2000) found contra-lateralized FPN activation during right
index finger tapping task (without visual cue), including the left
dorsal lateral premotor cortex and the left inferior parietal lobule. Moreover, Yuan et al. (2015) recently demonstrated that gait
velocity among cognitively normal older adults was significantly
associated with connectivity of the left-FPN. Therefore, given
that our study participants were all right-hand dominant, our
results are supported by the literature. We also extend the
current state of knowledge by using data generated from
a randomized controlled trial to demonstrate the potential
impact of aerobic exercise on FPN connectivity during right
hand finger tapping and the significant association between
FPN connectivity and mobility and cardiovascular capacity. An
alternative interpretation of these results may be that aerobic
exercise may help maintain mobility and cardiovascular capacity
among older adults with SIVCI via reducing cognitive load
(i.e., less FPN connectivity) required to perform less attention-
demanding motor task (i.e., dominant hand finger tapping). Our own previous work supports this latter concept (Hsu
et al., 2017). DISCUSSION Nevertheless,
we highlight the key strengths of our currents study design – a
randomized controlled trial – which are: (1) provides evidence
of causation; and (2) increased internal validity. Thus, our
study provides preliminary evidence to suggest that aerobic
exercise may impact functional connectivity in older adults A few limitations should be taken into consideration. First,
our study participants are likely healthier and to have superior
physical functioning than average older adults with mild SIVCI. This potential sample bias is somewhat unavoidable given the
requirement that participants be able to engage in progressive
AT safely. However, it also limits the generalizability of our
findings to the population of older adults with mild SIVCI
as a whole. Secondly, due to the small sample size of the
current study, the current dataset may not possess enough
power to detect small differences between the two groups. Provided that the study population is generally frail and older, the
occurrence of drop-out from potentially strenuous fMRI session
is to be expected. Future studies designed with larger sample
sizes are necessary to validate the notion of functional network
efficiency/inefficiency by providing sufficient power despite the
expectation of drop-out. Thirdly, it is possible that subsets of
pairwise connectivity between ROI within the FPN may have
driven the effects we observed; however, this was not further
investigated due to potential issue with type II error with the
current sample size. Moreover, there is much controversy in
regards to global signal regression and potential observation of
artificial anti-correlations. This may be particularly influential
when examining functional connectivity between networks
deemed anti-correlated in nature (e.g., default mode network
and FPN). In assessing within-network connectivity, it may be
that the effects of induced anti-correlation are less significant. However, as stated by Murphy and Fox (Murphy and Fox,
2016) ‘there is not a single “right” way to process resting state
data that reveals the “true” nature of the brain.’ They also
summarized the several advantages of global signal regression
including removal of motion, cardiac and respiratory signals. In addition, despite evidence supporting its use (Fair et al.,
2007; Zhu et al., 2017), we recognize temporally splicing and
concatenating data is not recommended and can potentially lead
to increase in signal noise. Nevertheless, studies demonstrated
that connectivity derived from concatenation does not differ
significantly from those acquired from continuous data (Fair
et al., 2007; Zhu et al., 2017). Frontiers in Human Neuroscience | www.frontiersin.org DISCUSSION Specifically,
Poppe et al. (2015) demonstrated that compared with healthy
controls, patients with schizophrenia had significantly less
connectivity in the FPN during goal-oriented task performance. Compared with controls, task performance was also significantly
worse among patients. Similarly, He et al. (2007) found that However, our results do concur with and extend emerging
evidence that show less functional connectivity of large-scale
networks may be advantageous (Chuang et al., 2014), especially
within the context of mobility. In one cross-sectional study,
Rosenberg-Katz et al. (2015) demonstrated that compared with FIGURE 3 | Correlation between change in TUG performance and change in FPN connectivity during right finger tapping within the AT group. orrelation between change in TUG performance and change in FPN connectivity during right finger tapping within the AT group. Frontiers in Human Neuroscience | www.frontiersin.org June 2017 | Volume 11 | Article 344 8 Understanding the Neural Underpinnings of Exercise Hsu et al. healthy older adults and individuals with Parkinson’s disease
who were non-fallers, those with Parkinson’s disease who were
fallers showed significantly greater connectivity between the
posterior parietal lobule and the inferior parietal lobule. This
data suggest increased connectivity between parietal regions
may be associated with more severe motor impairments and
more generally, heightened neural activity (e.g., activation
or connectivity) may reflect the inability of networks to
actively suppress irrelevant neural events, causing regions to
compete unnecessarily for available neural resources. In contrast,
diminished connectivity may represent greater efficiency as the
networks can effectively allocate resources to areas of immediate
importance. Certainly, emerging evidence suggests that lifestyle
interventions can improve neural efficiency (Smith et al., 2013;
Nishiguchi et al., 2015). network examined (default mode network vs. FPN); and (3) study
participant (healthy older adults vs. older adults with SIVCI). The
combination of these variations could have resulted in disparities
in the reported findings. in the reported findings. A few limitations should be taken into consideration. First,
our study participants are likely healthier and to have superior
physical functioning than average older adults with mild SIVCI. This potential sample bias is somewhat unavoidable given the
requirement that participants be able to engage in progressive
AT safely. However, it also limits the generalizability of our
findings to the population of older adults with mild SIVCI
as a whole. DISCUSSION Secondly, due to the small sample size of the
current study, the current dataset may not possess enough
power to detect small differences between the two groups. Provided that the study population is generally frail and older, the
occurrence of drop-out from potentially strenuous fMRI session
is to be expected. Future studies designed with larger sample
sizes are necessary to validate the notion of functional network
efficiency/inefficiency by providing sufficient power despite the
expectation of drop-out. Thirdly, it is possible that subsets of
pairwise connectivity between ROI within the FPN may have
driven the effects we observed; however, this was not further
investigated due to potential issue with type II error with the
current sample size. Moreover, there is much controversy in
regards to global signal regression and potential observation of
artificial anti-correlations. This may be particularly influential
when examining functional connectivity between networks
deemed anti-correlated in nature (e.g., default mode network
and FPN). In assessing within-network connectivity, it may be
that the effects of induced anti-correlation are less significant. However, as stated by Murphy and Fox (Murphy and Fox,
2016) ‘there is not a single “right” way to process resting state
data that reveals the “true” nature of the brain.’ They also
summarized the several advantages of global signal regression
including removal of motion, cardiac and respiratory signals. In addition, despite evidence supporting its use (Fair et al.,
2007; Zhu et al., 2017), we recognize temporally splicing and
concatenating data is not recommended and can potentially lead
to increase in signal noise. Nevertheless, studies demonstrated
that connectivity derived from concatenation does not differ
significantly from those acquired from continuous data (Fair
et al., 2007; Zhu et al., 2017). In addition, our data is limited
by the fact that only the connectivity during right hand tapping
was statistically significant while left hand was not. Differences in
social interactions experienced by the experimental groups may
present addition confounding factors to our data. Specifically,
active attention provided by trainers within the AT group
may potentially influence our findings. Lastly, the relationship
between connectivity and SIVCI status is equivocal with much
of the evidence generated from cross-sectional studies. Thus,
the inclusion of fMRI data from a healthy-aged matched cohort
might have facilitated interpretation of our results. REFERENCES Cataneo, D. C., Kobayasi, S., Carvalho, L. R., Paccanaro, R. C., and Cataneo,
A. J. (2010). Accuracy of six minute walk test, stair test and spirometry
using maximal oxygen uptake as gold standard. Acta Cir. Bras. 25, 194–200. doi: 10.1590/S0102-86502010000200013 Andrews-Hanna, J. R., Snyder, A. Z., Vincent, J. L., Lustig, C., Head, D., Raichle,
M. E., et al. (2007). Disruption of large-scale brain systems in advanced aging. Neuron 56, 924–935. doi: 10.1016/j.neuron.2007.10.038 Chuang, Y. F., Eldreth, D., Erickson, K. I., Varma, V., Harris, G., Fried, L. P.,
et al. (2014). Cardiovascular risks and brain function: a functional magnetic
resonance imaging study of executive function in older adults. Neurobiol. Aging Atkinson, H. H., Rosano, C., Simonsick, E. M., Williamson, J. D., Davis, C.,
Ambrosius, W. T., et al. (2007). Cognitive function, gait speed decline, and
comorbidities: the health, aging and body composition study. J. Gerontol. A Biol. Sci. Med. Sci. 62, 844–850. doi: 10.1093/gerona/62.8.844 35, 1396–1403. doi: 10.1016/j.neurobiolaging.2013.12.008 Sci. Med. Sci. 62, 844–850. doi: 10.1093/gerona/62.8.844 Cockrell, J. R., and Folstein, M. F. (1988). Mini-mental state examination (MMSE). Psychophar. Bull. 24, 689–692. Beckmann, C. F., Jenkinson, M., and Smith, S. M. (2003). General multilevel
linear modeling for group analysis in FMRI. Neuroimage 20, 1052–1063. doi: 10.1016/S1053-8119(03)00435-X Delbaere, K., Kochan, N. A., Close, J. C., Menant, J. C., Sturnieks, D. L., Brodaty, H.,
et al. (2012). Mild cognitive impairment as a predictor of falls in community-
dwelling older people. Am. J. Geriatr. Psychiatry 20, 845–853. doi: 10.1097/JGP. 0b013e31824afbc4 Blumen, H. M., Holtzer, R., Brown, L. L., Gazes, Y., and Verghese, J. (2014). Behavioral and neural correlates of imagined walking and walking-while-
talking in the elderly. Hum. Brain Mapp. 35, 4090–4104. doi: 10.1002/hbm. 22461 Desmond, D. W., Erkinjuntti, T., Sano, M., Cummings, J. L., Bowler, J. V.,
Pasquier, F., et al. (1999). The cognitive syndrome of vascular dementia:
implications for clinical trials. Alzheimer Dis. Assoc. Disord. 13(Suppl. 3),
S21–S29. doi: 10.1097/00002093-199912001-00005 Bolandzadeh, N., Tam, R., Handy, T. C., Nagamatsu, L. S., Hsu, C. L., Davis, J. C.,
et al. (2015). Resistance training and white matter lesion progression in older
women: exploratory analysis of a 12-month randomized controlled trial. J. Am. Geriatr. Soc. 63, 2052–2060. doi: 10.1111/jgs.13644 Ding, W., Cao, W., Wang, Y., Sun, Y., Chen, X., Zhou, Y., et al. (2015). Altered
functional
connectivity
in
patients
with
subcortical
vascular
cognitive
impairment–a
resting-state
functional
magnetic
resonance
imaging
study. PLoS
ONE
10:e0138180. FUNDING This work was supported by Canadian Stroke Network and the
Heart and Stroke Foundation of Canada to TL-A and the Jack
Brown and Family Alzheimer Research Foundation Society to
TL-A. DISCUSSION Critically, in this separate sub-analysis of the same
6-month RCT, we found that after AT, older adults confirmed
with SIVCI performed significantly better at the Eriksen flanker
task compared with the no-exercise controls. The observed
improvement in task performance was associated with overall
reduction in activation in the lateral occipital cortex and superior
temporal gyrus. It should be noted that we are aware of only one study in the
relevant field in the literature that investigated the association
between functional connectivity and cardiorespiratory fitness
among older adults (Voss et al., 2010a). While our findings
deviate from evidence presented, in which the authors reported
greater connectivity is associated with higher fitness among
healthy older adults (Voss et al., 2010a), several distinctions
from the current study should be considered. Specifically, the
differences were: (1) the fMRI task (visual vs. motor); (2) the June 2017 | Volume 11 | Article 344 9 Understanding the Neural Underpinnings of Exercise Hsu et al. AUTHOR CONTRIBUTIONS with SIVCI, and this is associated with the maintenance of
mobility. TL-A and RH were involved in the study concept, design,
acquisition of data, preparation and critical review of the
manuscript. CH, MM, WC, and TL-A were involved in data
collection. CH, TL-A, and JB were involved in writing of the
manuscript. CH, TL-A, and JB were involved in statistical
analyses. CH, TL-A, and JB were involved in interpretation of
data. CH and SW were involved in fMRI data analyses. MV and
TH were involved in critical review of the manuscript. All authors
had full access to all of the data (including statistical reports and
tables) in the study and can take responsibility for the integrity of
the data and the accuracy of the data analysis. CONCLUSION Our
results
demonstrate
that
neural
network
functional
connectivity may contribute to the effects of aerobic exercise
on mobility among older adults with SIVCI. We observed that
6 months of AT maintains motor task-based connectivity within
the FPN of older adults with SIVCI, and the degree of decoupling
within this region correlates with improvements in mobility. As such, our current findings support emerging results from
others that altered functional connectivity within certain neural
networks might represent a beneficial change in older adults with
mild SIVCI, especially vis-à-vis their mobility. More broadly,
these results bring further support to the burgeoning notion
that functional neural changes contribute to exercised-induced
improvements to mobility among older adults. As extension of
these findings, future studies should explore potential interaction
between mobility and cognitive outcomes among this population. ACKNOWLEDGMENT TL-A had full access to all the data in the study and takes
responsibility for the integrity of the data and the accuracy of the
data analysis. CH is an Alzheimer Society Research Program Doctoral trainee. TL-A is a Canada Research Chair (Tier II) in Physical Activity,
Mobility, and Cognitive Neuroscience. REFERENCES L., and
Corbetta, M. (2007). Breakdown of functional connectivity in frontoparietal
networks underlies behavioral deficits in spatial neglect. Neuron 53, 905–918. doi: 10.1016/j.neuron.2007.02.013 Odenheimer, G., Funkenstein, H. H., Beckett, L., Chown, M., Pilgrim, D., Evans, D.,
et al. (1994). Comparison of neurologic changes in ‘successfully aging’ persons
vs the total aging population. Arch. Neurol. 51, 573–580. doi: 10.1001/archneur. 1994.00540180051013 Holtzer, R., Mahoney, J. R., Izzetoglu, M., Izzetoglu, K., Onaral, B., and Verghese, J. (2011). fNIRS study of walking and walking while talking in young and old
individuals. J. Gerontol. A Biol. Sci. Med. Sci. 66, 879–887. doi: 10.1093/gerona/
glr068 Pahor, M., Guralnik, J. M., Ambrosius, W. T., Blair, S., Bonds, D. E., Church,
T. S., et al. (2014). Effect of structured physical activity on prevention of major
mobility disability in older adults: the LIFE study randomized clinical trial. JAMA 311, 2387–2396. doi: 10.1001/jama.2014.5616 Hsu, C. L., Best, J. R., Davis, J. C., Nagamatsu, L. S., Wang, S., Boyd, L. A., et al. (2017). Aerobic exercise promotes executive functions and impacts functional
neural activity among older adults with vascular cognitive impairment. Br J
Sports Med doi: 10.1136/bjsports-2016-096846 Pantoni, L., Leys, D., Fazekas, F., Longstreth, WT Jr, Inzitari, D., Wallin, A., et al. (1999). Role of white matter lesions in cognitive impairment of vascular origin. Alzheimer Dis. Assoc. Disord. 13(Suppl. 3), S49–S54. Parzen, E. (1962). On estimation of a probability density function and mode. Annal. Math. Stat. 33, 1065–1076. doi: 10.1214/aoms/1177704472 Hsu, C. L., Nagamatsu, L. S., Davis, J. C., and Liu-Ambrose, T. (2012). Examining
the relationship between specific cognitive processes and falls risk in older
adults: a systematic review. Osteoporos Int. 23, 2409–2424. doi: 10.1007/s00198-
012-1992-z Persinger, R., Foster, C., Gibson, M., Fater, D. C., and Porcari, J. P. (2004). Consistency of the talk test for exercise prescription. Med. Sci. Sports Exerc. 36,
1632–1636. doi: 10.1249/01.MSS.0000074670.03001.98 Hsu, C. L., Voss, M. W., Handy, T. C., Davis, J. C., Nagamatsu, L. S., Chan, A.,
et al. (2014). Disruptions in brain networks of older fallers are associated with
subsequent cognitive decline: a 12-month prospective exploratory study. PLoS
ONE 9:e93673. doi: 10.1371/journal.pone.0093673 Pool, E. M., Rehme, A. K., Fink, G. R., Eickhoff, S. B., and Grefkes, C. (2014). Handedness and effective connectivity of the motor system. Neuroimage 99,
451–460. doi: 10.1016/j.neuroimage.2014.05.048 Jancke, L., Loose, R., Lutz, K., Specht, K., and Shah, N. J. (2000). REFERENCES doi:
10.1371/journal.pone.013
8180 Borg, G. (1982). Ratings of perceived exertion and heart rates during short-term
cycle exercise and their use in a new cycling strength test. Int. J. Sports Med. 3,
153–158. doi: 10.1055/s-2008-1026080 Bowler, J. V. (2005). Vascular cognitive impairment. J. Neurol. Neurosurg. Psychiatry 76(Suppl. 5), v35–v44. doi: 10.1136/jnnp.2005.082313 Enright, P. L. (2003). The 6-min walk test: a quick measure of functional status in
elderly adults. Chest 123, 387–398. doi: 10.1378/chest.123.2.387 Buracchio, T., Dodge, H. H., Howieson, D., Wasserman, D., and Kaye, J. (2010). THe trajectory of gait speed preceding mild cognitive impairment. Arch. Neurol. 67, 980–986. doi: 10.1001/archneurol.2010.159 Erickson, K. I., and Kramer, A. F. (2009). Aerobic exercise effects on cognitive and
neural plasticity in older adults. Br. J. Sports Med. 43, 22–24. doi: 10.1136/bjsm. 2008.052498 Erkinjuntti, T., Bowler, J. V., Decarli, C. S., Fazekas, F., Inzitari, D., O’brien, J. T.,
et al. (1999). Imaging of static brain lesions in vascular dementia: implications
for clinical trials. Alzheimer Dis. Assoc. Disord. 13(Suppl. 3), S81–S90. doi: 10.1097/00002093-199912001-00013 Campbell, A. J., Robertson, M. C., Gardner, M. M., Norton, R. N., and Buchner,
D. M. (1999). Falls prevention over 2 years: a randomized controlled trial in
women 80 years and older. Age Ageing 28, 513–518. doi: 10.1093/ageing/28. 6.513 June 2017 | Volume 11 | Article 344 Frontiers in Human Neuroscience | www.frontiersin.org 10 Understanding the Neural Underpinnings of Exercise Hsu et al. improves executive functioning in older fallers: a randomized controlled
trial. J. Am. Geriatr. Soc. 56, 1821–1830. doi: 10.1111/j.1532-5415.2008. 01931.x Erkinjuntti, T., Inzitari, D., Pantoni, L., Wallin, A., Scheltens, P., Rockwood, K.,
et al. (2000). Research criteria for subcortical vascular dementia in clinical trials. J. Neural Transm. Suppl. 59, 23–30. doi: 10.1007/978-3-7091-6781-6_4 Liu-Ambrose, T., Eng, J. J., Boyd, L. A., Jacova, C., Davis, J. C., Bryan, S., et al. (2010). Promotion of the mind through exercise (PROMoTE): a proof-of-
concept randomized controlled trial of aerobic exercise training in older adults
with vascular cognitive impairment. BMC Neurol. 10:14. doi: 10.1186/1471-
2377-10-14 Fair, D. A., Schlaggar, B. L., Cohen, A. L., Miezin, F. M., Dosenbach, N. U.,
Wenger, K. K., et al. (2007). A method for using blocked and event-related fMRI
data to study “resting state” functional connectivity. Neuroimage 35, 396–405. doi: 10.1016/j.neuroimage.2006.11.051 Fogassi, L., and Luppino, G. (2005). Motor functions of the parietal lobe. Curr. Opin. Neurobiol. 15, 626–631. doi: 10.1016/j.conb.2005.10.015 McAusland, J., Tam, R., Wong, E., Riddehough, A., and Li, D. REFERENCES (2010). Optimizing
the use of radiologist seed points for improved multiple sclerosis lesion
segmentation. IEEE Trans. Biomed. Eng. 57, 2689–2698. doi: 10.1109/TBME. 2010.2055865 Fox, M. D., Zhang, D., Snyder, A. Z., and Raichle, M. E. (2009). The global signal
and observed anticorrelated resting state brain networks. J. Neurophysiol. 101,
3270–3283. doi: 10.1152/jn.90777.2008 Gao, Q., Wang, J., Yu, C., and Chen, H. (2015). Effect of handedness on brain
activity patterns and effective connectivity network during the semantic task
of Chinese characters. Sci. Rep. 5:18262. doi: 10.1038/srep18262 Montero-Odasso, M., Verghese, J., Beauchet, O., and Hausdorff, J. M. (2012). Gait
and cognition: a complementary approach to understanding brain function and
the risk of falling. J. Am. Geriatr. Soc. 60, 2127–2136. doi: 10.1111/j.1532-5415. 2012.04209.x Geerligs, L., Maurits, N. M., Renken, R. J., and Lorist, M. M. (2012). Reduced specificity of functional connectivity in the aging brain during task
performance. Hum. Brain Mapp. 35, 319–330. doi: 10.1002/hbm.22175 Murphy, K., and Fox, M. D. (2016). Towards a consensus regarding global signal
regression for resting state functional connectivity MRI. Neuroimage doi: 10. 1016/j.neuroimage.2016.11.052 Gellish, R. L., Goslin, B. R., Olson, R. E., Mcdonald, A., Russi, G. D., and Moudgil,
V. K. (2007). Longitudinal modeling of the relationship between age and
maximal heart rate. Med. Sci. Sports Exerc. 39, 822–829. doi: 10.1097/mss. 0b013e31803349c6 Nasreddine, Z. S., Phillips, N. A., Bedirian, V., Charbonneau, S., Whitehead, V.,
Collin, I., et al. (2005). The montreal cognitive assessment, MoCA: a brief
screening tool for mild cognitive impairment. J. Am. Geriatr. Soc. 53, 695–699. doi: 10.1111/j.1532-5415.2005.53221.x Groll, D. L., To, T., Bombardier, C., and Wright, J. G. (2005). The development of
a comorbidity index with physical function as the outcome. J. Clin. Epidemiol. 58, 595–602. doi: 10.1016/j.jclinepi.2004.10.018 Nishiguchi, S., Yamada, M., Tanigawa, T., Sekiyama, K., Kawagoe, T., Suzuki, M.,
et al. (2015). A 12-Week physical and cognitive exercise program can improve
cognitive function and neural efficiency in community-dwelling older adults: a
randomized controlled trial. J. Am. Geriatr. Soc. 63, 1355–1363. doi: 10.1111/
jgs.13481 Guralnik, J. M., Ferrucci, L., Simonsick, E. M., Salive, M. E., and Wallace, R. B. (1995). Lower-extremity function in persons over the age of 70 years as a
predictor of subsequent disability 10.1056/NEJM199503023320902. N. Engl. J. Med. 332, 556–562. doi: 10.1056/NEJM199503023320902 O’Brien, J. T. (2006). Vascular cognitive impairment. Am. J. Geriatr. Psychiatry 14,
724–733. doi: 10.1097/01.JGP.0000231780.44684.7e He, B. J., Snyder, A. Z., Vincent, J. L., Epstein, A., Shulman, G. REFERENCES 05.046 Rosenberg-Katz, K., Herman, T., Jacob, Y., Mirelman, A., Giladi, N., Hendler, T.,
et al. (2015). Fall risk is associated with amplified functional connectivity of the
central executive network in patients with Parkinson’s disease. J. Neurol. 262,
2448–2456. doi: 10.1007/s00415-015-7865-6 Wymbs, N. F., Bassett, D. S., Mucha, P. J., Porter, M. A., and Grafton, S. T. (2012). Differential recruitment of the sensorimotor putamen and frontoparietal cortex
during motor chunking in humans. Neuron 74, 936–946. doi: 10.1016/j.neuron. 2012.03.038 Seeley, W. W., Menon, V., Schatzberg, A. F., Keller, J., Glover, G. H., Kenna, H.,
et al. (2007). Dissociable intrinsic connectivity networks for salience processing
and executive control. J. Neurosci. 27, 2349–2356. doi: 10.1523/JNEUROSCI. 5587-06.2007 Yi, L., Wang, J., Jia, L., Zhao, Z., Lu, J., Li, K., et al. (2012). Structural and functional
changes in subcortical vascular mild cognitive impairment: a combined
voxel-based morphometry and resting-state fMRI study. PLoS ONE 7:e44758. doi: 10.1371/journal.pone.0044758 Shumway-Cook, A., Brauer, S., and Woollacott, M. (2000). Predicting the
probability for falls in community-dwelling older adults using the Timed Up
& Go Test. Phys. Ther. 80, 896–903. Yu, Y., Zhou, X., Wang, H., Hu, X., Zhu, X., Xu, L., et al. (2015). Small-world
brain network and dynamic functional distribution in patients with subcortical
vascular cognitive impairment. PLoS ONE 10:e0131893. doi: 10.1371/journal. pone.0131893 Smith, J. C., Nielson, K. A., Antuono, P., Lyons, J. A., Hanson, R. J., Butts, A. M.,
et al. (2013). Semantic memory functional MRI and cognitive function after
exercise intervention in mild cognitive impairment. J. Alzheimers. Dis. 37,
197–215. doi: 10.3233/JAD-130467 Yuan, J., Blumen, H. M., Verghese, J., and Holtzer, R. (2015). Functional
connectivity associated with gait velocity during walking and walking-while-
talking in aging: a resting-state fMRI study. Hum. Brain Mapp. 36, 1484–1493. doi: 10.1002/hbm.22717 Smith, S. M., Fox, P. T., Miller, K. L., Glahn, D. C., Fox, P. M., Mackay, C. E., et al. (2009). Correspondence of the brain’s functional architecture during activation
and rest. Proc. Natl. Acad. Sci. U.S.A. 106, 13040–13045. doi: 10.1073/pnas. 0905267106 Zhou, X., Hu, X., Zhang, C., Wang, H., Zhu, X., Xu, L., et al. (2016). Aberrant
functional connectivity and structural atrophy in subcortical vascular cognitive
impairment: relationship with cognitive impairments. Front. Aging Neurosci. 8:14. doi: 10.3389/fnagi.2016.00014 Sridharan, D., Levitin, D. J., and Menon, V. (2008). A critical role for the right
fronto-insular cortex in switching between central-executive and default-mode
networks. Proc. Natl. Acad. Sci. U.S.A. 105, 12569–12574. doi: 10.1073/pnas. REFERENCES Cortical
activations during paced finger-tapping applying visual and auditory pacing
stimuli. Brain Res. Cogn. Brain Res. 10, 51–66. doi: 10.1016/S0926-6410(00)00
022-7 Poppe, A. B., Carter, C. S., Minzenberg, M. J., and Macdonald, A. W. III. (2015). Task-based functional connectivity as an indicator of genetic liability to
schizophrenia. Schizophr. Res. 162, 118–123. doi: 10.1016/j.schres.2014.11.022 Robertson, M. C., Campbell, A. J., Gardner, M. M., and Devlin, N. (2002). preventing injuries in older people by preventing falls: a meta-analysis of
individual-level data. J. Am. Geriatr. Soc. 50, 905–911. doi: 10.1046/j.1532-5415. 2002.50218.x Kuo, H.-K., and Lipsitz, L. A. (2004). Cerebral white matter changes and geriatric
syndromes: is there a link? J. Gerontol. A Biol. Sci. Med. Sci. 59, M818–M826. doi: 10.1093/gerona/59.8.m818 Rockwood, K., Wentzel, C., Hachinski, V., Hogan, D. B., Macknight, C.,
and Mcdowell, I. (2000). Prevalence and outcomes of vascular cognitive
impairment. Vascular cognitive impairment investigators of the canadian study
of health and aging. Neurology 54, 447–451. doi: 10.1212/WNL.54.2.447 Layne, A. S., Hsu, F. C., Blair, S. N., Chen, S. H., Dungan, J., Fielding, R. A.,
et al. (2017). Predictors of change in physical function among older adults in
response to long-term, structured physical activity: the LIFE Study. Arch. Phys. Med. Rehabil. 98, 11.e3–24.e3. doi: 10.1016/j.apmr.2016.07.019 Liu-Ambrose, T., Nagamatsu, L. S., Hsu, C. L., and Bolandzadeh, N. (2013). Emerging concept: ’central benefit model’ of exercise in falls prevention. Br. J. Sports Med. 47, 115–117. doi: 10.1136/bjsports-2011-090725 Roman, G. C., Erkinjuntti, T., Wallin, A., Pantoni, L., and Chui, H. C. (2002). Subcortical ischaemic vascular dementia. Lancet Neurol. 1, 426–436. doi: 10. 1016/S1474-4422(02)00190-4 Liu-Ambrose, T., Best, J. R., Davis, J. C., Eng, J. J., Lee, P. E., Jacova, C., et al. (2016). Aerobic exercise and vascular cognitive impairment: a randomized controlled
trial. Neurology 87, 2082–2090. doi: 10.1212/WNL.0000000000003332 Rosano, C., Newman, A. B., Katz, R., Hirsch, C. H., and Kuller, L. H. (2008). Association between lower digit symbol substitution test score and slower gait
and greater risk of mortality and of developing incident disability in well-
functioning older adults. J. Am. Geriatr. Soc. 56, 1618–1625. doi: 10.1111/j. 1532-5415.2008.01856.x Liu-Ambrose, T., Donaldson, M. G., Ahamed, Y., Graf, P., Cook, W. L.,
Close, J., et al. (2008). Otago home-based strength and balance retraining June 2017 | Volume 11 | Article 344 Frontiers in Human Neuroscience | www.frontiersin.org 11 Understanding the Neural Underpinnings of Exercise Hsu et al. Parkinson’s disease. Neurosci. Lett. 460, 6–10. doi: 10.1016/j.neulet.2009. June 2017 | Volume 11 | Article 344 Frontiers in Human Neuroscience | www.frontiersin.org REFERENCES 0800005105 Zhu, Y., Cheng, L., He, N., Yang, Y., Ling, H., Ayaz, H., et al. (2017). Comparison
of functional connectivity estimated from concatenated task-state data from
block-design paradigm with that of continuous task. Comput. Math. Methods
Med. 2017:4198430. doi: 10.1155/2017/4198430 Vermeer, S. E., Longstreth, W. T. Jr., and Koudstaal, P. J. (2007). Silent brain
infarcts: a systematic review. Lancet Neurol. 6, 611–619. doi: 10.1016/S1474-
4422(07)70170-9 Voss, M. W., Erickson, K. I., Prakash, R. S., Chaddock, L., Malkowski, E., Alves, H.,
et al. (2010a). Functional connectivity: a source of variance in the association
between
cardiorespiratory
fitness
and
cognition? Neuropsychologia
48,
1394–1406. doi: 10.1016/j.neuropsychologia.2010.01.005 Conflict of Interest Statement: The authors declare that the research was
conducted in the absence of any commercial or financial relationships that could
be construed as a potential conflict of interest. Voss, M. W., Prakash, R. S., Erickson, K. I., Basak, C., Chaddock, L., Kim, J. S.,
et al. (2010b). Plasticity of brain networks in a randomized intervention trial of
exercise training in older adults. Front. Aging Neurosci. 2:32. doi: 10.3389/fnagi. 2010.00032 The reviewers KM, GA and handling Editor declared their shared affiliation, and
the handling Editor states that the process nevertheless met the standards of a fair
and objective review. Washburn, R. A., Mcauley, E., Katula, J., Mihalko, S. L., and Boileau, R. A. (1999). The physical activity scale for the elderly (PASE): evidence for validity. J. Clin. Epidemiol. 52, 643–651. doi: 10.1016/S0895-4356(99)00049-9 Copyright © 2017 Hsu, Best, Wang, Voss, Hsiung, Munkacsy, Cheung, Handy
and Liu-Ambrose. This is an open-access article distributed under the terms of
the Creative Commons Attribution License (CC BY). The use, distribution or
reproduction in other forums is permitted, provided the original author(s) or licensor
are credited and that the original publication in this journal is cited, in accordance
with accepted academic practice. No use, distribution or reproduction is permitted
which does not comply with these terms. Wise, S. P., Boussaoud, D., Johnson, P. B., and Caminiti, R. (1997). Premotor and
parietal cortex: corticocortical connectivity and combinatorial computations. Annu. Rev. Neurosci. 20, 25–42. doi: 10.1146/annurev.neuro.20.1.25 Wu, T., Wang, L., Chen, Y., Zhao, C., Li, K., and Chan, P. (2009). Changes
of functional connectivity of the motor network in the resting state in June 2017 | Volume 11 | Article 344 Frontiers in Human Neuroscience | www.frontiersin.org 12
|
https://openalex.org/W3172108632
|
https://www.nature.com/articles/s42003-021-02170-6.pdf
|
English
| null |
An Oligocene giant rhino provides insights into Paraceratherium evolution
|
Communications biology
| 2,021
|
cc-by
| 8,599
|
1 Key Laboratory of Vertebrate Evolution and Human Origins, Institute of Vertebrate Paleontology and Paleoanthropology, Chinese Academy of Sciences,
Beijing, China. 2 CAS Center for Excellence in Life and Paleoenvironment, Beijing, China. 3 University of Chinese Academy of Sciences, Beijing, China. 4 Henan
University of Chinese Medicine, Zhengzhou, Henan, China. 5 Department of Human Evolutionary Biology, Harvard University, Cambridge, MA, USA.
6 Hezheng Paleozoological Museum, Hezheng, Gansu, China. ✉email: dengtao@ivpp.ac.cn COMMUNICATIONS BIOLOGY | (2021) 4:639 | https://doi.org/10.1038/s42003-021-02170-6 | www.nature.com/commsbio An Oligocene giant rhino provides insights into
Paraceratherium evolution T p
y
Here we report a completely preserved skull with an articulated
mandible and atlas, representing a new species of Para-
ceratherium from the upper Oligocene Jiaozigou Formation of the
Linxia Basin in Gansu Province, China, located at the north-
eastern border of the Tibetan Plateau11,12. The Jiaozigou fauna of
the Linxia Basin also includes the giant rhinos Turpanotherium
and Dzungariotherium, the rodent Tsaganomys, the creodont
Megalopterodon, the chalicothere Schizotherium, the hyracodont
Ardynia, the rhinocerotid Aprotodon, and the entelodont Para-
entelodon (Supplementary Table 1), making it similar to the
Nanpoping fauna of the Lanzhou Basin and other Tabenbulukian
faunas from Inner Mongolia and Ningxia. This suggests the
widespread occurrence of open woodland during the late Oligo-
cene in northwestern China, with a mix of woodland and
grassland7. Comparative description. The new species differs from other
species of Paraceratherium (P. grangeri, P. huangheense, P. asiaticum, P. bugtiense, and P. lepidum) in having a deeper nasal
notch whose bottom is located above the middle of M2, pro-
portionally larger height of the condyle (43.9%) compared to
height of the occipital surface (Supplementary Table 2), short
muzzle bones and diastema anterior to cheek teeth, highly raised
occiput, and high zygomatic arch with a prominent posterior end
(Fig. 1). The nasals of P. linxiaense are flat and straight, and the nasal
notch is very deep above the middle of M2 with a short distance
from the orbit (15.3% of the basal cranial length) (Supplementary
Table
2),
much
deeper
than
those
of
other
species
of
Paraceratherium except P. lepidum, indicating a short prehensile
nose trunk. The dorsal surface of the skull is shallowly depressed,
different from the domed skull of P. grangeri13 or the flat one of
P. lepidum7. The distance between the parietal crests is narrow
and smaller than that of P. lepidum. The infraorbital foramen is
situated above the P4/M1 boundary, and the anterior margin of
the orbit is located above the M2/M3 boundary. Both characters
are similar to those of P. lepidum and more posteriorly positioned
than those of P. grangeri above the P3/P4 boundary and the
middle of M2 respectively. The position of the zygomatic arch is
high, posteriorly flush with the upper margin of the orbit like
those of P. grangeri and P. lepidum, and much higher than that of
P. bugtiense14. The postorbital process is absent. An Oligocene giant rhino provides insights into
Paraceratherium evolution Paraceratherium linxiaense possesses features that
characterize the genus, such as a giant body size, long premaxillae
with anterior ends extending downward, separated parietal crests,
high condyle compared to the height of nuchal surface, lower
inferior border of the posttympanic process than the condyle,
roughly horizontal anterior part of symphysis, and downward
turning cone-shaped I1. It is more derived than other species
within this genus in having a larger body size, deeper nasal notch
above M2, much higher occipital part and posterior end of
zygomatic arch, and smaller upper incisor I1. The lower margin
of the horizontal mandibular ramus is concave under the dia-
stema, and small i1 extends anteriorly and horizontally. The
dental formula is 1.0.3.3/1.0.3.3. P2 is semimolarized, while P3
and P4 are submolarized. The metaconule connects with the
ectoloph and the anterior point of the hypocone in moderate
wear; the antecrochet is moderate; the lingual border of the
protocone is rounded on molars; and the ecto-posterior corner of
the protolophid is angular on p3 and p4. The atlas has an
expanded
transverse
foramen
and
a
dumb-bell
shaped
vertebral fossa. T
he giant rhino (derived genera of Paraceratheriidae, Rhi-
nocerotoidea, Perissodactyla, such as Paraceratherium,
Dzungariotherium, Aralotherium, and Turpanotherium)
has been considered as one of the largest land mammals that ever
lived1. Its skull and legs are longer than all reported land mam-
mals, but the metapodials are not massive in outline. Its body size
was suitable for open woodlands under humid or arid climatic
conditions2. Except for some remains found in Eastern Europe3,
Anatolia4,5, and Caucasus6, giant rhinos lived mainly in Asia,
especially in China, Mongolia, Kazakhstan, and Pakistan7. All
forms of the giant rhino, including six genera, have been recorded
from northwest to southwest China through the middle Eocene to
the late Oligocene. Among them, Juxia from the middle Eocene is
considered as the unequivocal ancestor of all giant rhinos because
of a set of the primitive and primarily specialized features7. The
genus Paraceratherium was the most widely distributed form of
the giant rhino, but aside from East and Central Asia, many
records from East Europe3 and West Asia4–6 comprise frag-
mentary specimens, and only Paraceratherium bugtiense, known
from the southwestern corner of the Tibetan Plateau8, has ample
records and undoubtable taxa identity and is key to the origin and
dispersal history of Paraceratherium9,10. An Oligocene giant rhino provides insights into
Paraceratherium evolution The space
between the posttympanic and postglenoid processes is moderate
as in P. lepidum, wider than in P. grangeri and narrower than in
P. bugtiense. The posttympanic process has no transverse
expansion, and its lateral margin is almost flush with the
postglenoid process. The posttympanic and paraoccipital pro-
cesses fuse to become a wide and thick plate. An Oligocene giant rhino provides insights into
Paraceratherium evolution Tao Deng
1,2,3✉, Xiaokang Lu4, Shiqi Wang1,2, Lawrence J. Flynn5, Danhui Sun1,3, Wen He6 & Shanqin Chen6 Tao Deng
1,2,3✉, Xiaokang Lu4, Shiqi Wang1,2, Lawrence J. Flynn5, Danhui Sun1,3, Wen He6 & Shanqin Chen6 As one of the largest land mammals, the origin and evolution of the giant rhino Para-
ceratherium bugtiense in Pakistan have been unclear. We report a new species Paraceratherium
linxiaense sp. nov. from northwestern China with an age of 26.5 Ma. Morphology and phy-
logeny reveal that P. linxiaense is the highly derived species of the genus Paraceratherium, and
its clade with P. lepidum has a tight relationship to P. bugtiense. Based on the paleogeo-
graphical literature, P. bugtiense represents a range expansion of Paraceratherium from Central
Asia via the Tibetan region. By the late Oligocene, P. lepidum and P. linxiaense were found in
the north side of the Tibetan Plateau. The Tibetan region likely hosted some areas with low
elevation, possibly under 2000 m during Oligocene, and the lineage of giant rhinos could
have dispersed freely along the eastern coast of the Tethys Ocean and perhaps through some
lowlands of this region. 1 ARTICLE COMMUNICATIONS BIOLOGY | https://doi.org/10.1038/s42003-021-02170-6 Diagnosis. Paraceratherium linxiaense possesses features that
characterize the genus, such as a giant body size, long premaxillae
with anterior ends extending downward, separated parietal crests,
high condyle compared to the height of nuchal surface, lower
inferior border of the posttympanic process than the condyle,
roughly horizontal anterior part of symphysis, and downward
turning cone-shaped I1. It is more derived than other species
within this genus in having a larger body size, deeper nasal notch
above M2, much higher occipital part and posterior end of
zygomatic arch, and smaller upper incisor I1. The lower margin
of the horizontal mandibular ramus is concave under the dia-
stema, and small i1 extends anteriorly and horizontally. The
dental formula is 1.0.3.3/1.0.3.3. P2 is semimolarized, while P3
and P4 are submolarized. The metaconule connects with the
ectoloph and the anterior point of the hypocone in moderate
wear; the antecrochet is moderate; the lingual border of the
protocone is rounded on molars; and the ecto-posterior corner of
the protolophid is angular on p3 and p4. The atlas has an
expanded
transverse
foramen
and
a
dumb-bell
shaped
vertebral fossa. Diagnosis. COMMUNICATIONS BIOLOGY | https://doi.org/10.1038/s42003-021-02170-6 Fig. 1 Holotype (HMV 2006) of Paraceratherium linxiaense sp. nov. Skull: a lateral view; b ventral view; c dorsal view; d anterior view; e occipital view
Mandible: f, h lateral view and medial view of left ramus, respectively; g occlusal view. Skull and mandible share the scale bar, but both anterior and nucha
views have an independent scale bar. Fig. 1 Holotype (HMV 2006) of Paraceratherium linxiaense sp. nov. Skull: a lateral view; b ventral view; c dorsal view; d anterior view; e occipital view. Mandible: f, h lateral view and medial view of left ramus, respectively; g occlusal view. Skull and mandible share the scale bar, but both anterior and nuchal
views have an independent scale bar. The distance between the anterior margins of I1 and P2 is 164
mm. DP1 is absent like in most species of Paraceratherium, but
present in P. grangeri. The paracone rib is absent from P2 to M1,
weak in M2, and marked in M3, which is the common character
of the genus Paraceratherium, much different from the strong
paracone rib of the primitive giant rhino Forstercooperia15. The
occlusal surface of P2 is triangular in P. linxiaense, different from
the trapezium outline in P. grangeri and P. asiaticum. There is an
obvious separating groove between the protocone and hypocone
of P3 and P4. The hypocone of P4 is situated behind the
metaconule as in P. grangeri and P. lepidum, and it is expanded
and rounded, while that of P. lepidum is square. The antecrochet
on upper molars is larger than those of P. grangeri and P. asiaticum. M3 is triangular in occlusal outline, similar to those of
P. bugtiense and P. lepidum, but different from the rectangular
outline in P. grangeri and P. asiaticum. Eocene to early Oligocene Aralotherium and the late Oligocene
Dzungariotherium and Turpanotherium, and among the species
within the genus Paraceratherium has not been revealed through
cladistics analysis. y
Our phylogenetic analysis of 155 characters, including 73
newly added cranial, dental, and postcranial characters, coded
from 11 giant rhinos and 16 other Rhinocerotoidea taxa
(including 2 extant), yield a single most parsimonious tree with
length 327, consistency index 0.60, and retention index 0.79
(Fig. 6), which places P. linxiaense as a derived giant rhino,
nested
within
the
monophyletic
clade
of
the
Oligocene
Asian Paraceratherium. In the most parsimonious trees (Sup-
plementary Figs. COMMUNICATIONS BIOLOGY | https://doi.org/10.1038/s42003-021-02170-6 1, 2), Paraceratherium has a relationship closer
to late Eocene Aralotherium than to Dzungariotherium and
Turpanotherium7. The latter two late Oligocene genera formed a
clade. Like Paraceratherium, Aralotherium continues the ten-
dency to enlarge the lower incisor i1 of the giant rhino to the
maximum level, but its incisor is implanted in an anteriorly
downward bent symphysis, which is derived from the straight
stage seen in Urtinotherium and Paraceratherium. The most
outstanding morphology of the late Eocene Aralotherium, as well
the largest giant rhino Dzungariotherium lies in having a nasal
notch extending posteriorly to over and above the anterior rim of
the orbit. This extremely specialized nasal notch is more specific
to the giant rhino. Within Rhinocerotoidea, evidently, Para-
ceratherium is at a moderate level for this specialization, which is
further demonstrated by the less derived cheek teeth and con-
servative postcranial bones. Nevertheless, it remains uncertain
whether the relatively smaller body size and incisors in Ara-
lotherium and Turpanotherium are retained from the primitive
ancestor or reduced from larger paraceratheres. g
g
P. linxiaense has more reduced i1. The p2 is small and single-
rooted, but large and double-rooted in P. grangeri. The p3 and p4
have
a
rudimentary
entolophid,
while
the
entoconid
of
P. asiaticum is an isolated cusp. The entolophid of m3 is nearly
transverse. Additional descriptions, measurements and comparisons, such
as information of the postcranial bones, are provided in
Supplementary Note 2 and Supplementary Tables 1–6. Results Results
Systematic paleontology. Perissodactyla Owen, 1848
Paraceratheriidae Osborn, 1923
Paraceratherium Forster-Cooper, 1911
Paraceratherium linxiaense sp. nov. Type specimens. A complete skull and mandible with the asso-
ciated atlas (holotype, HMV 2006, Fig. 1), and an axis and two
thoracic vertebrae of another individual (paratype, HMV 2007,
Fig. 2), which are preserved at the Hezheng Paleozoological
Museum in Hezheng County, Gansu Province, China. HMV 2006
represents a full adult individual. The specific name, linxiaense,
refers to the geographical location of the discovery in the Linxia
Basin (Fig. 3). For P. linxiaense, the posterior border of the mandibular
symphysis is situated at the p4/m1 boundary, and the posterior
margin of the ascending ramus is anteriorly inclined, different
from vertical situation in P. bugtiense and P. lepidum. The
vascular notch of lower margin of the mandible is deep but
more anterior than that of P. lepidum. The mandibular
diastema has a straight and slowly declining upper margin like
in P. grangeri, while strongly declined in P. asiaticum and
P. lepidum, and convex in P. huangheense and P. bugtiense. The
mental foramen is situated under the p3/p4 boundary, more
posteriorly than in P. asiaticum, P. bugtiense, and P. lepidum
where is under p2. Type locality and horizon. IVPP locality LX1808 (N35°35’05.16”,
E103°18’51.02”; 1983 m above sea level, Fig. 4) is near the village
of Wangjiachuan, 10.8 km southwest of the town of Dongxiang
County, Linxia Hui Autonomous Prefecture, Gansu Province,
China (Fig. 3). HMV 2006 and 2007 are from the sandstones in
the lower part of the Jiaozigou Formation (Fig. 5, Supplementary
Note 1). Age. IVPP locality LX1808 is faunally and paleomagnetically
dated to the middle of chron C8r with an estimated age of 26.5
million years ago (Ma) in the late Oligocene (Fig. 5). COMMUNICATIONS BIOLOGY | (2021) 4:639 | https://doi.org/10.1038/s42003-021-02170-6 | www.nature.com/commsbio 2 ARTICLE COMMUNICATIONS BIOLOGY | (2021) 4:639 | https://doi.org/10.1038/s42003-021-02170-6 | www.nature.com/commsbio Discussion In addition,
the latter two species have a much larger body, for example, the
mandibular length of P. bugtiense is 4/5 of P. lepidum7 or P. lin-
xiaense. P. lepidum and P. linxiaense have different directions of
specialization. Within Paraceratherium, upturning of the lower
margin of the symphysis has a tendency of gradually backward
reduction. It is at the level of p2 in P. grangeri and P. huangheense, at
p3
in
P. bugtiense
and
P. linxiaense,
but
at
p4
in
P. lepidum7,17–19. Adaptation of the atlas and axis to the large body
and long neck of the giant rhino already characterized P. grangeri
and P. bugtiense, and was further developed in P. linxiaense, whose
atlas is elongated, indicative of a long neck and higher axis with a
nearly horizontal position of its posterior articular face. There
correlate to a more flexible neck17,19–21. Paraceratherium lepidum
has an atlas resembling that of P. bugtiense, and an axis resembling
that of P. linxiaense. In terms of limb bones, P. lepidum has the
most massive third metacarpal within the genus. P. grangeri, through P. huangheense, P. asiaticum, and P. bugtiense,
and terminating in P. lepidum and P. linxiaense (Fig. 6) (see
Methods and Supplementary Data 1–3 for all details of the ana-
lysis). Paraceratherium linxiaense exhibits a high level of speciali-
zation, similar to P. lepidum, and both the concerned clade and P. bugtiense derive from a common ancestor (P. linxiaense has slender
skull and cervical vertebrae). This hypothesis is strongly supported
by the morphology of mandible. A notch on the lower margin of
the mandible in front of the mandibular angle occurred firstly in P. bugtiense, where it is shallow and concave, and then evolved as a
deeper and wider notch in P. linxiaense and P. lepidum. In addition,
the latter two species have a much larger body, for example, the
mandibular length of P. bugtiense is 4/5 of P. lepidum7 or P. lin-
xiaense. P. lepidum and P. linxiaense have different directions of
specialization. Within Paraceratherium, upturning of the lower
margin of the symphysis has a tendency of gradually backward
reduction. It is at the level of p2 in P. grangeri and P. huangheense, The giant rhino of western Pakistan is from the Oligocene pre-
Siwalik Chitarwata Formation. Discussion With the previous studies7,16 as a basis, the systematics of para-
ceratheres has been partially resolved at the genus level. These
works suggest that Juxia diverged from the Forstercooperia-Pap-
pacera clade at about 40 Ma in the middle Eocene, and its stock
evolved into Urtinotherium in the late Eocene and the derived
genus Paraceratherium in the Oligocene. However, the phyloge-
netic relationships among other derived genera, including the late Within the Paraceratherium clade, our phylogenetic analysis
produces a series of progressively more-derived species from 3 MMUNICATIONS BIOLOGY | (2021) 4:639 | https://doi.org/10.1038/s42003-021-02170-6 | www.nature.com/commsbio ARTICLE COMMUNICATIONS BIOLOGY | https://doi.org/10.1038/s42003-021-02170-6 Fig. 2 Vertebrae of Paraceratherium linxiaense. a atlas (HMV 2006); b axis (HMV 2007); c 4th–5th articulated thoracic vertebrae (HMV 2007). 1, right
view; 2, posterior view; 3, left view; 4, dorsal view; 5, anterior view; 6, ventral view; 7, postero-ventral view. The articular facets on 4th-5th thoracic
vertebrae are marked with the red semitransparent outline whatever it is intact or has been damaged. Fig. 2 Vertebrae of Paraceratherium linxiaense. a atlas (HMV 2006); b axis (HMV 2007); c 4th–5th articulated t
view; 2, posterior view; 3, left view; 4, dorsal view; 5, anterior view; 6, ventral view; 7, postero-ventral view. The
vertebrae are marked with the red semitransparent outline whatever it is intact or has been damaged. Fig. 2 Vertebrae of Paraceratherium linxiaense. a atlas (HMV 2006); b axis (HMV 2007); c 4th–5th articulated thoracic vertebrae (HMV 2007). 1, right
view; 2, posterior view; 3, left view; 4, dorsal view; 5, anterior view; 6, ventral view; 7, postero-ventral view. The articular facets on 4th-5th thoracic
vertebrae are marked with the red semitransparent outline whatever it is intact or has been damaged. P. grangeri, through P. huangheense, P. asiaticum, and P. bugtiense,
and terminating in P. lepidum and P. linxiaense (Fig. 6) (see
Methods and Supplementary Data 1–3 for all details of the ana-
lysis). Paraceratherium linxiaense exhibits a high level of speciali-
zation, similar to P. lepidum, and both the concerned clade and P. bugtiense derive from a common ancestor (P. linxiaense has slender
skull and cervical vertebrae). This hypothesis is strongly supported
by the morphology of mandible. A notch on the lower margin of
the mandible in front of the mandibular angle occurred firstly in P. bugtiense, where it is shallow and concave, and then evolved as a
deeper and wider notch in P. linxiaense and P. lepidum. Discussion Specimens represent a single
species, Paraceratherium bugtiense, while the genus distributed
across
the
Mongolian
Plateau,
northwestern
China,
and
Kazakhstan north to the Tibetan Plateau is highly diversified. Our phylogenetic analysis indicates that all six species of Para-
ceratherium are sister to Aralotherium and form a clade in which COMMUNICATIONS BIOLOGY | (2021) 4:639 | https://doi.org/10.1038/s42003-021-02170-6 | www.nature.com/commsbio 4 ARTICLE COMMUNICATIONS BIOLOGY | https://doi.org/10.1038/s42003-021-02170-6 Fig. 3 Type locality of Paraceratherium linxiaense. Map showing the giant rhino fossil locality of the Linxia Basin in Wangjiachuan Village, Dongxiang
County, Gansu Province, China. Fig. 3 Type locality of Paraceratherium linxiaense. Map showing the giant rhino fossil locality of the Linxia Basin in Wangjiachuan Village, Dongxiang
County, Gansu Province, China. County, Gansu Province, China. Fig. 4 Type horizon of Paraceratherium linxiaense. Exposures of fluvial and lacustrine sediments of the Linxia Basin (LX 1808) at the type locality of this
new species (HMV 2006, yellow square) and where the axis and thoracic vertebrae were also found in Wangjiachuan Village, Dongxiang County, Gansu
Province, China. Fig. 4 Type horizon of Paraceratherium linxiaense. Exposures of fluvial and lacustrine sediments of the Linxia Basin (LX 1808) at the type locality of this
new species (HMV 2006, yellow square) and where the axis and thoracic vertebrae were also found in Wangjiachuan Village, Dongxiang County, Gansu
Province, China. still not established as a high-elevation plateau. The largest land
mammal might have been able to migrate freely, passing along
eastern coast of the Tethys in the western part of Tibet, and even
through some lowlands within what is now the plateau. P. grangeri is the most primitive, succeeded by P. huangheense
and P. asiaticum. In the early Oligocene, P. asiaticum dispersed
westward to Kazakhstan from the hypothesized ancestral area of
the genus Paraceratherium in Mongolia where P. grangeri origi-
nated, and its descendant lineage might have expanded to South
Asia as P. bugtiense. In the late Oligocene, as the sister group of P. bugtiense, P. lepidum was found in Ningxia, Xinjiang and
Kazakhstan, and P. linxiaense in Linxia. Our phylogenetic ana-
lysis places P. bugtiense of South Asia as a derivative of Central
Asian P. huangheense via P. asiaticum, indicating dispersal
southward through the Tibetan region. We note early Oligocene
aridity in Central Asia at a time when South Asia was relatively
moist, with a mosaic of forested and open landscapes22,23. COMMUNICATIONS BIOLOGY | (2021) 4:639 | https://doi.org/10.1038/s42003-021-02170-6 | www.nature.com/commsbio Discussion Late
Oligocene tropical conditions allowed the giant rhino to return
northward to Central Asia, implying that the Tibetan region was P. grangeri is the most primitive, succeeded by P. huangheense
and P. asiaticum. In the early Oligocene, P. asiaticum dispersed
westward to Kazakhstan from the hypothesized ancestral area of
the genus Paraceratherium in Mongolia where P. grangeri origi-
nated, and its descendant lineage might have expanded to South
Asia as P. bugtiense. In the late Oligocene, as the sister group of P. bugtiense, P. lepidum was found in Ningxia, Xinjiang and
Kazakhstan, and P. linxiaense in Linxia. Our phylogenetic ana-
lysis places P. bugtiense of South Asia as a derivative of Central
Asian P. huangheense via P. asiaticum, indicating dispersal
southward through the Tibetan region. We note early Oligocene
aridity in Central Asia at a time when South Asia was relatively
moist, with a mosaic of forested and open landscapes22,23. Late
Oligocene tropical conditions allowed the giant rhino to return
northward to Central Asia, implying that the Tibetan region was The paracerathere evolutionary trend can be analyzed against
geological time. P. grangeri appeared in the Mongolian Plateau
during the late early Oligocene with an age of about 31–28 Ma at
Hsanda Gol, Mongolia24, and it produced westward derivatives P. huangheense and P. asiaticum, reaching the Lanzhou Basin and
the Turgai region in Kazakhstan, respectively19,25. P. bugtiense
comes from the Oligocene Bugti Member of the lower Chitarwata
Formation in South Asia9,10,26. Our cladistic analysis places
the Oligocene P. bugtiense as sister taxon to the P. lepidum and
P. linxiaense dichotomy. Late Oligocene northward dispersal
of these two new forms is recorded as widespread fossils of 5 MMUNICATIONS BIOLOGY | (2021) 4:639 | https://doi.org/10.1038/s42003-021-02170-6 | www.nature.com/commsbio ARTICLE COMMUNICATIONS BIOLOGY | https://doi.org/10.1038/s42003-021-02170-6 COMMUNICATIONS BIOLOGY | https://doi.org/10.1038/s42003-021-02170-6 m distributed in the vast region between the Mongolian
7
The southwest depression of the Tarim Basin ex
zoic section of the Linxia Basin. Deposits with correlation to ATNTS 201249 (left) for the magnetostratigraphic results50 (m
an section bearing the fossils of Paraceratherium linxiaense sp. nov. (right). Fig. 5 Cenozoic section of the Linxia Basin. Deposits with correlation to ATNTS 201249 (left) for the magnetostratigraphic results50 (middle) and the
Wangjiachuan section bearing the fossils of Paraceratherium linxiaense sp. nov. (right). P. lepidum distributed in the vast region between the Mongolian
Plateau and Kazakhstan7, and by P. linxiaense currently known
only in the Linxia Basin. Discussion The southwest depression of the Tarim Basin extended east-
ward to reach the Altyn Tagh Mountain where the late Eocene
to the early Miocene marine deposits are distributed and bear
abundant marine foraminifers, ostracods, and bivalves28–30. Marine fish fossils, among which cartilaginous fishes strongly
resemble the fish fauna of the Paris Basin31, indicate an east-
west sea in this area (Supplementary Note 3 and Supplemen-
tary Fig. 3). South Asia was separated from Kazakhstan by the
sea, blocking giant rhinos from direct exchange in this direc-
tion (Fig. 7). The southwest depression of the Tarim Basin extended east-
ward to reach the Altyn Tagh Mountain where the late Eocene
to the early Miocene marine deposits are distributed and bear
abundant marine foraminifers, ostracods, and bivalves28–30. Marine fish fossils, among which cartilaginous fishes strongly
resemble the fish fauna of the Paris Basin31, indicate an east-
west sea in this area (Supplementary Note 3 and Supplemen-
tary Fig. 3). South Asia was separated from Kazakhstan by the
sea, blocking giant rhinos from direct exchange in this direc-
tion (Fig. 7). y
Since the giant rhinos of Kazakhstan and Pakistan were
contemporaries in the Oligocene, could they disperse directly
between the two regions? Paleogeographically, it was impos-
sible. In the Eocene, Asia was separated from Europe because
the Middle East and Western Siberia were still occupied by the
sea27. The Turgai Strait was still submerged but transitional
terrestrial deposits were already developing in the Oligocene25. COMMUNICATIONS BIOLOGY | (2021) 4:639 | https://doi.org/10.1038/s42003-021-02170-6 | www.nature.com/commsbio 6 ARTICLE COMMUNICATIONS BIOLOGY | https://doi.org/10.1038/s42003-021-02170-6 Fig. 6 Phylogenetic relationship of giant rhinos. Correlated with the geographical and geochronological distribution, based on the single most-
parsimonious tree (length 327, consistency index 0.60, and retention index 0.79, with Bremer support values nearby the node). Fig. 6 Phylogenetic relationship of giant rhinos. Correlated with the geographical and geochronological distribution, based on the single most-
parsimonious tree (length 327, consistency index 0.60, and retention index 0.79, with Bremer support values nearby the node). Fig. 7 Distribution and dispersal of Paraceratherium. Localities of the early Oligocene species were marked by the yellow color, and the red indicates the
late Oligocene species. Dispersals of Paraceratherium between South Asia and other localities have to pass the Tibetan region, because most part of Centra
Asia, including southeastern Kazakhstan, Turpan Basin, and Tarim Basin was covered by the Tethys Ocean during the Oligocene. COMMUNICATIONS BIOLOGY | https://doi.org/10.1038/s42003-021-02170-6 During the Oligocene, dispersal for the giant rhino from the
Mongolian Plateau to South Asia could have been along the
eastern coast of the Tethys like other mammals, such as
anthracotheres32 and ruminants33 as early as in the late Eocene or
around the Eocene-Oligocene transition, or through some low-
altitude areas in the hinterland of Tibet, demonstrating that the
Tibetan region was not yet an elevated plateau as it is today,
without height great enough to obstruct the dispersal of large
mammals like the giant rhino. The topographical possibility that
the giant rhino may have passed through the Tibetan region to
reach the Indian-Pakistani subcontinent in the Oligocene can be
supported by other evidence. The fish and plant fossils discovered
from the Oligocene lacustrine deposits in the central Tibetan
region display tropical characteristics, indicating an elevation of
lower than 2000 m for this region34–36. Through to the late
Oligocene, the evolution and dispersal from P. bugtiense to P. linxiaense and P. lepidum demonstrate that Tibet, as a plateau,
did not yet exist and was not yet a barrier to exchange of largest
land mammals. Phylogeny of Hyracodontidae has long been debated but remains hardly
resolved. Many authors hypothesized a closer relationship of Hyracodontidae and
Paraceratheriidae7,15,37,45, and several assigned it as a stem group basal to other
rhinocerotids16,42,43,46. In this analysis, however, Triplopus was presented as a stem
group sister to a “hyracodontid clade” of three genera Epitriplopus, Hyracodon, and
Triplopides, which is not supported by postcranial features but is by four dental
features (node B in Fig. 6): 63(1), lower canine is reduced to incisor size; 68(2),
hypocone and protocone on P2 isolated and not connected with each other; 71(1),
hypocone and protocone on P4 isolated but connected; 76(1), posterior part of the
ectoloph on M1–2 is straight. The failure of this study to find a monophyletic
hyracodontid group is not too surprising, due to its unresolved relationship with
Uintaceras, Pappaceras and Forstercooperia (the latter two taxa were united with
the paraceratheriids), as well as the limited taxa and postcranial bones of
hyracodontid here for this family. The relationship of Hyracodontidae is therefore
tentative and should be reassessed in the future studies. Methods
N
l Nomenclature Acts. This published work and the nomenclatural acts have been
registered in ZooBank, the proposed online registration system for the Interna-
tional Code of Zoological Nomenclature (ICZN). The ZooBank LSIDs (Life Science
Identifiers) can be resolved and the associated information viewed through any
standard web browser by appending the LSID to the prefix “http://zoobank.org/”. The LSID for this publication is: urn:lsid:zoobank.org:pub:04E1C419-F1C1-4876-
ACE9-3DC199F03696. The similarities of cheek teeth and skulls between Forstercooperia and Juxia
have been discussed several times7,14,48. Our result did not show a monophyly of
Forstercooperiinae, but supported a close relationship of Pappaceras and
Forstercooperia, and Juxia with derived giant rhinos (node D in Fig. 6), these
genera share four synapomorphies (9(1), zygomatic arch with a lobe-shaped blade;
69(0), widest position of P3-P4 crown at middle part; 97(1) and 98(1), trigonid of
lower molar with a nearly flat or round out wall, and nearly U-shaped outline in
occlusal view). Among Paleogene rhinocerotoids, the lobe-shaped blade of
zygomatic arch is only present on groups in this clade. The reversal of P3-P4
outline should affect the phylogeny of less specialized groups of rhinocerotoid, such
as Forstercooperia, Triplopus, and Uintaceras, but discussion of this issue is beyond
the scope of this paper. Statistics and Reproducibility. Based on the many studies on giant rhinos,
especially the detailed discussion and recommendations7, we consider Dzungar-
iotherium and Aralotherium as valid genera, but Indricotherium and Baluchither-
ium as junior synonyms of Paraceratherium. There were several revisions at species
level7,14,21. Benaratherium is not taken into account in this study because its
limited materials are not accessible. In this first phylogeny analysis of Para-
ceratheriidae, we selected all 6 species of Paraceratherium and the type species of
all other genera of this family7, with the exception of Turpanotherium, which is
represented by T. yagouense skull material instead of the less well represented type
species T. elegans. Other Paleogene rhinocerotoid sister taxa, including two genera
of Amynodontidae, four genera of Hyracodontidae and four of Rhinocerotidae,
were added to test their relationship with Paraceratheriidae. In particular, with
respect to Rhinocerotidae the extant species, plus 3 genera from Neogene and
Quaternary were used to further reveal trends. Hyrachyus was used as outgroup. Whenever possible, we selected taxa with well-preserved samples and distinct
diagnoses and morphology, especially taxa previously used in phylogenetic analyses
to make well-tested comparisons. Methods
N
l The samples of each taxon used in this study to
code the characters were listed in Supplementary Data 1. The true giant rhino clade (node E in Fig. 6) was supported by thirteen
synapomorphies (1(2), nasal notch retracted; 4(1), premaxilla and nasal contact
absent; 12(1), infraorbital foramen below nasal notch; 32(1), mental foramen at
level of p2–p4; 39(1), upper incisor I1 with large size; 44(1), upper incisors
sagittally arranged; 50(1), upraised extension of lower incisor i1 absent; 51(1),
downward extension of lower incisor i1; 52(1), lower incisor i1 enlarged; 63(1),
lower canine reduced in size; 68(2), hypocone and protocone isolated and not
connected with each other in P2; 69(2), widest position of P3-P4 at posterior part
of crown; 70(1), hypocone and protocone isolated but connected with each other in
P3), with strong Bremer support value of 5 steps. Among these synapomorphies,
the shape of nose, premaxilla, and lower and upper first incisors, make the giant
rhino differ from all other Paleogene rhinocerotoids as well as Forstercooperiinae
(Char. 1, 4, 12, 50, 51, 52), and the posteriorly situated widest position on P3-P4
(Char. 69) has never been observed on either Paleogene or later rhinocerotoids. On
node F (Fig. 6), our analysis put a feature that represents a typical evolutionary
tendency of Paraceratheriidae, namely the highly specialized chisel-liked lower
incisor i1 (54(2), 61(1)), which united Urtinotherium and derived giant rhinos,
together with another synapomorphies (28 (1)). Large body size is the most outstanding feature of Paraceratheriidae, especially
in the derived genera. In this study, we described and defined 73 new characters
based on careful investigations and comparisons of giant rhinos, particularly the
postcranial bones of derived paraceratheriids, including 11 of cranium and
mandible, 22 of teeth, and 40 of postcranial bones. Additionally, we used the
previously built 82 characters to cover the morphology of other Paleogene
rhinoceroses and derived rhinoceros list groups, but with new data set and
descriptions16,37–40. All characters used in this study appear in Supplementary
Data 2. In total, 155 characters were scored for the 27 taxa (Supplementary Data 3), all
equally weighted and non-additive. The phylogenetic analysis was performed using
TNT 1.5 in traditional search with the strategy of WAG and TBR, 100 random
seed41 (Supplementary Fig. 1). In order to obtain all the most parsimonious trees,
the analysis was performed with 1000 replications, each of which got the best score. COMMUNICATIONS BIOLOGY | https://doi.org/10.1038/s42003-021-02170-6 Contrary to the recent previous analysis44, our analysis of Rhinocerotidae found
a closer relationship with Paraceratheriidae, sharing four synapomorphies (14(1),
postglenoid foramen absent; 32(1), mental foramen at level of p2–p4; 110(1),
robust humerus; 128(1), proximal articular facet in McIII high and evident). The
significance of the absence of postglenoid foramen in the phylogeny of both
families has been highlighted14. The posteriorly retracted mental foramen in the
synapomorphies must be correlated with the enlarged lower incisor and
mandibular symphysis, although each has different incisor specialization. This
relationship of families partly reflects the ample postcranial samples of the taxa. The enlarged incisors of Rhinocerotidae had not yet appeared in Uintaceras, which
therefore was not referred to this family although often has been considered as the
closest primitive sister group39,47. It shares four synapomorphies with
Rhinocerotidae clade (node C in Fig. 6) (24(1), occipital condyle with a median
ridge; 83(1), M1-M2 with antecrochet; 114(0), shorter radius and ulnar relative to
humerus; 155(1), MtIV with higher position relative to MtIII). The latter two
characters listed in this node illustrate a unique evolution pattern to robust legs, not
yet observed in other rhinocerotoids, shedding new light on the phylogeny of
Rhinocerotidae. Discussion Paleogeography map is
modified from Deep Time Maps (https://deeptimemaps.com) with license and a recent study51. Fig. 7 Distribution and dispersal of Paraceratherium. Localities of the early Oligocene species were marked by the yellow color, and the red indicates the
late Oligocene species. Dispersals of Paraceratherium between South Asia and other localities have to pass the Tibetan region, because most part of Central
Asia, including southeastern Kazakhstan, Turpan Basin, and Tarim Basin was covered by the Tethys Ocean during the Oligocene. Paleogeography map is
modified from Deep Time Maps (https://deeptimemaps.com) with license and a recent study51. 7 7 COMMUNICATIONS BIOLOGY | (2021) 4:639 | https://doi.org/10.1038/s42003-021-02170-6 | www.nature.com/commsbio MMUNICATIONS BIOLOGY | (2021) 4:639 | https://doi.org/10.1038/s42003-021-02170-6 | www.nature.com/commsbio ARTICLE ARTICLE COMMUNICATIONS BIOLOGY | https://doi.org/10.1038/s42003-021-02170-6 7. Qiu, Z.-X. & Wang, B.-Y. Paracerathere fossils of China. Pal. Sin. N. Ser. 29,
1–396 (2007). alternative interpretation for the morphological differences among these genera is
that the loss of incisors and p1-p2 derived from the further reduction of the rostral
region, that is to say, Paraceratherium is probably a sister group of the ancestor of
Aralotherium, Dzungariotherium, and Turpanotherium. Dzungariotherium has the
largest body size, most reduced rostral teeth, shortened diastema and symphysis. In
Aralotherium, the most outstanding feature is the downward extension of lower
incisor i1, and its body size is smaller than Paraceratherium and 8. Antoine, P.-O. et al. New remains of the baluchithere Paraceratherium
bugtiense (Pilgrim, 1910) from the late/latest Oligocene of the Bugti Hills,
Balochistan, Pakistan. J. Asian Earth Sci. 24, 71–77 (2004). g
g
g
Balochistan, Pakistan. J. Asian Earth Sci. 24, 71–77 (2004). 9. Antoine, P.-O. et al. Early rhinocerotids (Mammalia, Perissodactyla) from
South Asia and a review of the Holarctic Paleogene rhinocerotid record. Can. J. Earth Sci. 40, 365–374 (2003). y
Dzungariotherium. But probably the smallest genus is Turpanotherium whose
rostral area is also reduced but with less reduced i1. It should be noted that this pair
of sister clades were united based on another hypothesis. The reduced i1 and
smaller body size is not a reversal, but probably derived from a less specialized or
enlarged lineage, particularly for a family whose distinct tendency is gigantism. 10. Métais, G. et al. Lithofacies, depositional environments, regional
biostratigraphy and age of the Chitarwata Formation in the Bugti H
Balochistan, Pakistan. J. Asian Earth Sci. 34, 154–167 (2009). 10. Métais, G. et al. Lithofacies, depositional environments, regional
biostratigraphy and age of the Chitarwata Formation in the Bugti Balochistan, Pakistan. J. Asian Earth Sci. 34, 154–167 (2009) 11. Deng, T., Qiu, Z.-X., Wang, B.-Y., Wang, X.-M. & Hou, S.-K. In Fossil
Mammals of Asia: Neogene Biostratigraphy and Chronology (eds. Wang, X.-
M., Flynn, L. J. & Fortelius, M.) 243–273 (Columbia Uni. Press, New York,
2013). ARTICLE & Sobus, J. In The Evolution of Perissodac 5. Radinsky, L. B. A review of the rhinocerotoid family Hyraco 15. Radinsky, L. B. A review of the rhinocerotoid family Hyracod
(Perissodactyla). Bull. Am. Mus. Nat. Hist. 136, 1–45 (1967). (Perissodactyla). Bull. Am. Mus. Nat. Hist. 136, 1–45 (19 16. Wang, H.-B., Bai, B., Meng, J. & Wang, Y.-Q. Earliest known unequivocal
rhinocerotoid sheds new light on the origin of giant rhinos and phylogeny of
early rhinocerotoids. Sci. Rep. 6, 39607 (2016). 17. Forster-Cooper, C. On the skull and dentition of Paraceratherium bugtiense, a
genus of aberrant rhinoceros from the lower Miocene deposits of Dera Bugti. Philos. Trans. Roy. Soc. Lond. Ser. B 212, 369–394 (1924). y
18. Pavlow, M. Indricotherium transouralicum, n. sp. provenant du district de
Tourgay. Bull. Soc. Nat. Mosc. N. Ser. 31, 95–116 (1922). Tourgay. Bull. Soc. Nat. Mosc. N. Ser. 31, 95–116 (1922). 19. Li, Y.-X., Zhang, Y.-X., Li, J., Li, Z.-C. & Xie, K. New fossils of paraceratheres
(Perissodactyla, Mammalia) from the early Oligocene of the Lanzhou Basin,
Gansu Province, China. Vert. PalAsiat. 56, 367–381 (2017). 20. Granger, W. & Gregory, W. K. Further notes on the gigantic extinct
rhinoceros Baluchitherium from the Oligocene of Mongolia. Bull. Am. Mus. Nat. Hist. 72, 1–73 (1936). Based on the same matrix (27 taxa and 155 characters in Supplementary
Data 3), we also run the likelihood estimation of relationship of the giant rhino,
using Mrbayes 3.2.7 (lset nst = 6, rates = invgamma, ngen = 10000000). Within
clade of Paraceratherium, six species display a relationship similar to the result of
parsimonious analysis, but the relationships among four late Oligocene genera of
the giant rhino are not resolved. Relationship of the Paleogene rhinocerotoids is
also ambiguous in the likelihood estimation (Supplementary Fig. 2). 21. Gromova, V. Giant rhinoceroses (in Russian). Trav. Pal. Inst. Acad. Sci. USSR
71, 1–164 (1959). 22. De Franceschi, D. et al. Floral data from the mid-Cenozoic of central Pakistan. Rev. Palaeobot. Palyno. 150, 115–129 (2008). 23. Martin, C., Bentaleb, I. & Antoine, P.-O. Pakistan mammal tooth stable
isotopes show paleoclimatic and paleoenvironmental changes since the early
Oligocene. Palaeogeogr. Palaeoclimatol. Palaeoecol. 311, 19–29 (2011). Reporting summary. Further information on research design is available in the Nature
Research Reporting Summary linked to this article. 24. Höck, V. et al. Oligocene–Miocene sediments, fossils and basalts from the
Valley of Lakes (Central Mongolia): an integrated study. Mitt. Geol. Ges. ARTICLE 90,
83–125 (1999). 25. Lucas, S. G., Kordikova, E. G. & Emry, R. J. Oligocene stratigraphy, sequence
stratigraphy, and mammalian biochronology north of the Aral Sea, western
Kazakhstan. Bull. Carnegie Mus. Nat. Hist. 34, 313–348 (1998). Data availability All specimens (HMV 2006 and HMV 2007) are deposited at the Hezheng
Paleozoological Museum in Hezheng County, Gansu Province, China. Supporting data
(character list and data matrix) for phylogenetic analyses in this study are provided in
Supplementary Information. 26. Welcomme, J.-L. et al. Himalayan forelands: palaeontological evidence for
Oligocene detrital deposits in the Bugti Hills (Balochistan, Pakistan). Geol. Mag. 138, 397–405 (2001). 27. Wang, P.-X. Cenozoic deformation and the history of sea-land interactions in
Asia. AGU Geophys. Monogr. Ser. 149, 1–22 (2004). Received: 21 February 2020; Accepted: 29 April 2021; Received: 21 February 2020; Accepted: 29 April 2021; 28. Ritts, B. D. et al. From sea level to high elevation in 15 million years: uplift
history of the northern Tibetan Plateau margin in the Altun Shan. Am. J. Sci. 308, 657–678 (2008). 29. Wei, H.-H., Meng, Q.-R., Ding, L. & Li, Z.-Y. Tertiary evolution of the western
Tarim Basin, northwest China: a tectono-sedimentary response to northward
indentation of the Pamir salient. Tectonics 32, 558–575 (2013). Methods
N
l A single most parsimonious tree was attained with length 327, CI 0.60, RI 0.79
(Fig. 6). Bremer support values were calculated also by TNT with the TBR in
10 steps. Other than basal Juxia and Urtinotherium, the similarities shared by all derived
Paraceratheriidae are the fully reduced of anterior teeth even the first lower
premolars, these features are listed in node G (Fig. 6) (58(1), i2 absent; 60(1), i3
absent; 62(1), canine absent; 88(1), p1 absent). Following these changes are two
clades with more diverse morphology. One consists of Dzungariotherium and
Turpanotherium, and is supported by six synapomorphies (node H in Fig. 6) (13
(2), skull dorsal profile concave; 76(1), posterior part of ectoloph of M1–2 straight;
78(1), lingual cingulum of M1-M2 absent; 80(1), protocone constriction of M1-M2
remarkable; 81(1), hypocone constriction of M1-M2 present; 83(1), antecrochet of
M1-M2 present). Another clade (node I) (Paraceratherium and Aralotherium) is
united by two synapomorphies (53(2), highly enlarged i1; 57(0), enlarged i1 on
both sides distant). Based on known materials, these differences between the four
genera were enumerated7, with which our results are partially consistent in terms
of tendency of specialization: complicated occlusal pattern of cheek teeth of node
H, and presence of enlarged incisors of node I (Fig. 6). There are other features not
listed in the node but attributed to shape the different evolution patterns, which
separated these two clades, such as the reduced first upper or lower incisors in
Dzungariotherium and Turpanotherium, and absence of p2 in the latter. The Relationships of the family Paraceratheriidae. Amynodontids were once con-
sidered as a sister group to rhinocerotids or paraceratheriids in previous
work16,42,43. In our analysis, the amynodontid clade (Amynodon and Ros-
triamynodon) (node A in Fig. 6) is basal, following the outgroup Hyrachyus, with
high Bremer support values (4) and four synapomorphies (1(1), shallow nasal
notch anterior to P2; 47(2) and 63(2), highly enlarged upper and lower canines; 88
(1), absence of the first lower premolar p1), the latter three at greatly derived level
make it differ from all other Paleogene rhinocerotoids. This result corresponds to
the suggested Amynodontidae phylogeny37,44. COMMUNICATIONS BIOLOGY | (2021) 4:639 | https://doi.org/10.1038/s42003-021-02170-6 | www.nature.com/commsbio 8 ARTICLE g
g
p
y
y
y
g g
Paceratherium clade is supported by eleven synapomorphies (node J) (2(1), nasal
notch retracted to P2-P3; 6(1), length of diastema longer than upper premolar; 13
(1), arched skull dorsal profile; 29(1), robust crest along diastema of symphysis; 55
(1), very strong cingulum in i1; 95(1), entoconid connected with hypoconid in p4;
130(1), posterior McII facet in McIII present; 131(1), posterior McIV facet in McIII
absent; 139(1), astragalus trochlea extended downward; 145(1), outline of second
calcaneus facet in astragalus rounded; 155(1), higher position of MtIV relative to
MtIII. This result highlighted many unique features of Paraceratherium relative to
other derived giant rhinos, such as rostral area (Char. 6, 29), proximal articular facet
in McIII (Char. 130, 131), outline of astragalus (Char. 139, 145), but further
supported reversals in several species and their phylogenetic relationship within
Paraceratherium suggested in the above section of this study. P. grangeri appeared
as the ancestral morphotype succeeded by the second node (K) including P. huangheense whose lower cheek teeth were less specialized and four other species,
with two synapomorphies (83(1), antecrochet in M1-M2 present; 90(1), single root
in lower premolar p2). The remaining species were placed in the third node (L) with
three synapomorphies (67(1), much smaller size of P2 relative to P3-P4; 76(1),
straight posterior part of ectoloph in M1-M2; 80(1), remarkable protocone
constriction in M1-M2). P. asiaticum is more derived than P. grangeri in having a
gradually isolated hypocone in P3-P4, but less derived than P. bugtiense because of
the presence of upper premolar P1. P. bugtiense, P. lepidum and P. linxiaense
comprised node M, with two synapomorphies (64(1), upper premolar P1 absent; 82
(1), remarkable hypocone constriction in M1-M2). The node N is supported by two
synapomorphies (34(1), coronoid process of ramus little developed; 85(1), crochet
in M1-M2 present. These last two species represent the most specialized lineage
with large body size and complicated occlusal pattern of the upper cheek teeth. 12. Fang, X.-M. et al. Tectonosedimentary evolution model of an intracontinental
flexural (foreland) basin for paleoclimatic research. Glob. Planet. Change 145,
78–97 (2016). 13. Osborn, H. F. Baluchitherium grangeri, a giant hornless rhinoceros from
Mongolia. Am. Mus. Novit. 78, 1–15 (1923). g
14. Lucas, S. G. & Sobus, J. In The Evolution of Perissodactyls (eds. Prothero, D. R. & Schoch, R. M.) 358–378 (Oxford Univ. Press, New York, 1989). g
14. Lucas, S. G. COMMUNICATIONS BIOLOGY | (2021) 4:639 | https://doi.org/10.1038/s42003-021-02170-6 | www.nature.com/commsbio Additional information Supplementary information The online version contains supplementary material
available at https://doi.org/10.1038/s42003-021-02170-6. g
44. Heissig, K. In The Evolution of Perissodactyls (eds. Prothero, D. R. & Schoch,
R. M.) 399–417 (Oxford Univ. Press, New York, 1989). 45. Prothero, D. R. & Schoch, R. M. In The Evolution of Perissodactyls (eds. Prothero, D. R. & Schoch, R. M.) 504–529 (Oxford Univ. Press, New York,
1989). Correspondence and requests for materials should be addressed to T.D. Correspondence and requests for materials should be addressed to T.D. Reprints and permission information is available at http://www.nature.com/reprints Reprints and permission information is available at http://www.nature.com/reprints 46. Wang, H.-B., Bai, B., Meng, J. & Wang, Y.-Q. A new species of Forstercooperia
(Perissodactyla: Paraceratheriidae) from northern China with a systematic
revision of forstercooperiines. Am. Mus. Novit. 3897, 1–42 (2018). Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in
published maps and institutional affiliations. Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in
published maps and institutional affiliations. 47. Prothero, D. R. The Evolution of North American Rhinoceroses (Cambridge
Univ. Press, Cambridge, 2005). 48. Chow, M.-C., Chang, Y.-P. & Ting, S.-Y. Some Early Tertiary Perissodactyla
from Lunan Basin, E. Yunnan. Vert. PalAsiat. 12, 262–278 (1974). Open Access This article is licensed under a Creative Commons
Attribution 4.0 International License, which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give
appropriate credit to the original author(s) and the source, provide a link to the Creative
Commons license, and indicate if changes were made. The images or other third party
material in this article are included in the article’s Creative Commons license, unless
indicated otherwise in a credit line to the material. If material is not included in the
article’s Creative Commons license and your intended use is not permitted by statutory
regulation or exceeds the permitted use, you will need to obtain permission directly from
the copyright holder. To view a copy of this license, visit http://creativecommons.org/
licenses/by/4.0/. 49. Hilgen, F. J., Lourens, L. J. & Van Dam, J. A. In The Geologic Time Scale 2012
(eds. Gradstein, F. M., Ogg, J. G., Schmitz, M. K. & Ogg, G. M.) 923–978
(Elsevier, Amsterdam, 2012). 50. Fang, X.-M., Garzione, C., Van der Voo, R., Li, J.-J. & Fan, M.-J. Flexural
subsidence by 29 Ma on the NE edge of Tibet from the magnetostratigraphy of
Linxia Basin, China. Additional information Earth Planet. Sci. Lett. 210, 545–560 (2003). 51. Spicer, R. A. et al. Why ‘the uplift of the Tibetan Plateau’ is a myth. Natl Sci. Rev. https://doi.org/10.1093/nsr/nwaa091 (2020). ARTICLE COMMUNICATIONS BIOLOGY | https://doi.org/10.1038/s42003-021-02170-6 W. Zhang for taking surface scanning. We thank R. Blakey for providing the Oligocene
paleogeographical map, which was modified in Fig. 7 and Supplementary Fig. 3. Prof. Robert Spicer, Prof. Pierre-Olivier Antoine, and an anonymous reviewer are kindly
acknowledged for their comments on the manuscript. This research was supported by the
Chinese Academy of Sciences (XDB26000000, XDA20070203, QYZDY-SSW-DQC022,
GJHZ1885), the National Natural Science Foundation of China (41430102), and the
Second Comprehensive Scientific Expedition on the Tibetan Plateau (2019QZKK0705). 36. Botsyun, S. et al. Revised paleoaltimetry data show low Tibetan Plateau
elevation during the Eocene. Science 363, 946 (2019). 37. Prothero, D. R., Manning, E. & Hanson, C. B. The phylogeny of the
Rhinocerotoidea (Mammalia, Perissodactyla). Zool. J. Linn. Soc. 87, 341–366
(1986). 37. Prothero, D. R., Manning, E. & Hanson, C. B. The phylogeny of the Rhinocerotoidea (Mammalia, Perissodactyla). Zool. J. Linn. Soc. 87, 341–366
(1986). 38. Cerdeño, E. Cladistic analysis of the Family Rhinocerotidae (Perissodactyla). Am. Mus. Novit. 3143, 1–25 (1995). 39. Holbrook, L. T. & Lucas, S. G. A new genus of rhinocerotoid from the Eocene
of Utah and the status of North American “Forstercooperia”. J. Vert. Paleont. 17, 384–396 (1997). Author contributions T.D. designed the research project; T.D., S.W., W.H., and S.C. conducted the fieldwork;
T.D., X.L., and D.S. performed the phylogenetic analyses; T.D., X.L., and L.J.F. recon-
structed paleogeography; and T.D., X.L. and L.J.F. wrote the manuscript. 40. Antoine, P.-O. Phylogénie et évolution des Elasmotheriina (Mammalia
Rhinocerotidae). Mém. Mus. Natl Hist. Nat. 188, 1–359 (2002). oine, P.-O. Phylogénie et évolution des Elasmotheriina (Mammal 41. Goloboff, P. A., Farris, J. S. & Nixon, K. C. TNT, a free program for
phylogenetic analysis. Cladistics 24, 774–786 (2008). phylogenetic analysis. Cladistics 24, 774–786 (2008). Competing interests 42. Holbrook, L. T. The phylogeny and classification of tapiromorph
perissodactyls (Mammalia). Cladistics 15, 331–350 (1999). The authors declare no competing interests. 43. Tissier, J. et al. New data on Amynodontidae (Mammalia, Perissodactyla)
from Eastern Europe: phylogenetic and palaeobiogeographic implications
around the Eocene-Oligocene transition. Plos ONE 13, e0193774 (2018). References 30. Bosboom, R. et al. Timing, cause and impact of the late Eocene stepwise sea
retreat from the Tarim Basin (west China). Palaeogeogr. Palaeoclimatol. Palaeoecol. 403, 101–118 (2014). 1. Fortelius, M. & Kappelman, J. The largest land mammal ever imagined. Zool. J. Linn. Soc. 107, 85–101 (1993). 1. Fortelius, M. & Kappelman, J. The largest land mammal ever imagined. Zool. J. Linn. Soc. 107, 85–101 (1993). 2. Prothero, D. R. Rhinoceros Giants, the Paleobiology of Indricotheres (Indiana
Univ. Press, Bloomington, 2013). 2. Prothero, D. R. Rhinoceros Giants, the Paleobiology of Indricotheres (Indiana
Univ. Press, Bloomington, 2013). 31. Li, G.-Q. Elasmobranchs from the Lower Tertiary of western Tarim Basin,
China, and their biostratigraphic significance. Palaeoworld 7, 107–136 (1997). 3. Spassov, N. B. On the taxonomic status of Indricotherium Borissiak and giant
rhinocerotoids: indricotheres (Perissodactyla). C. R. Acad. Bulg. Sci. 42, 61–64
(1989). 32. Böhme, M. et al. Na Duong (northern Vietnam): an exceptional window into
Eocene ecosystems from Southeast Asia. Zitteliana 53, 120–167 (2013). 33. Métais, G., Mennecart, B. & Roohi, G. A new assemblage of stem pecoran
ruminants from the Oligocene Chitarwata Formation, Bugti Hills, Baluchistan,
Pakistan: Paleoenvironmental and paleobiogeographic implications. J. Asian
Earth Sci. 136, 40–49 (2017). 4. Antoine, P.-O., Karadenizli, L., Saraç, G. & Sen, S. A giant rhinocerotoid
(Mammalia, Perissodactyla) from the late Oligocene of north-central Anatolia
(Turkey). Zool. J. Linn. Soc. 152, 581–592 (2008). (Turkey). Zool. J. Linn. Soc. 152, 581–592 (2008). 5. Sen, S., Antoine, P.-O., Varol, B., Ayyildiz, T. & Sözeri, K. Giant rhinoceros
Paraceratherium and other vertebrates from Oligocene and middle Miocene
deposits of the Kağızman-Tuzluca Basin, Eastern Turkey. Naturwissenschaften
98, 407–423 (2011). 34. Wu, F.-X., Miao, D.-S., Chang, M.-M., Shi, G.-L. & Wang, N. Fossil climbing
perch and associated plant megafossils indicate a warm and wet central Tibet
during the late Oligocene. Sci. Rep. 7, 878 (2017). g
g
p
35. Su, T. et al. No high Tibetan Plateau until the Neogene. Sci. Adv. 5, eaav2189
(2019). 6. Gabunia, L. K. La Faune de Mammifères de l’Oligocène du Benara (Mezniereba
Press, Tbilisi, 1964). 9 9 COMMUNICATIONS BIOLOGY | (2021) 4:639 | https://doi.org/10.1038/s42003-021-02170-6 | www.nature.com/commsbio © The Author(s) 2021 Acknowledgements We thank Z.-X. Qiu, B.-Y. Wang, Z.-D. Qiu, X.-M. Wang, and S.-K. Hou for discussion,
X.-M. Fang for help in the field, Y. Chen for drawing, W. Gao for photographing, and © The Author(s) 2021 10 COMMUNICATIONS BIOLOGY | (2021) 4:639 | https://doi.org/10.1038/s42003-021-02170-6 | www.nature.com/commsbio
|
https://openalex.org/W2008555978
|
https://europepmc.org/articles/pmc4174445?pdf=render
|
English
| null |
Determination of in vivo RNA kinetics using RATE-seq
|
RNA
| 2,014
|
cc-by
| 6,980
|
INTRODUCTION cally result in a stress response or cellular death (Nonet et al. 1987) resulting in the estimation of mRNA decay rates that
may have little physiological relevance. Remodeling of gene expression is critical for a broad range of
biological processes from the cell division cycle and embryo
development (Schier 2007) to cellular responses to extracel-
lular signals (Gasch et al. 2000). Regulation of transcript
abundance is controlled by the combined action of transcript
synthesis and transcript degradation. Although the regulation
of transcript synthesis has historically been the primary focus
of investigation, there is accumulating evidence that RNA
degradation plays an important role in dynamic biological
processes (Elkon et al. 2010). A comprehensive understand-
ing of the regulation of gene expression programs, and the
development of mathematical models that explain the dy-
namics of gene expression, requires the accurate estimation
of absolute rates of both RNA synthesis and RNA degradation
in vivo. Recently, methods using in vivo metabolic labeling of
mRNAs (Cleary et al. 2005; Dölken et al. 2008) have been in-
troduced using either the nucleobase 4-thiouracil (4tU) or
nucleoside 4-thiouridine (4sU), which introduce a reactive
thiol group into RNAs. Following RNA purification, the pres-
ence of a thiol group in RNAs enables conjugation to N-
[6-(Biotinamido)hexyl]-3′-(2′-pyridyldithio)-propionamide
(biotin-HPDP) and subsequent fractionation using streptavi-
din-coated magnetic beads. Genome-wide estimation of in
vivo kinetic parameters using metabolic labeling of RNA
with 4tU has been reported using different experimental de-
signs. Pulse-chase labeling with 4tU (Munchel et al. 2011)
represents a promising approach to estimating mRNA degra-
dation rates. However, internal recycling of labeled nucleo-
tides (Puckett et al. 1975; Nikolov and Dabeva 1985) may
result in an incomplete chase thereby confounding the esti-
mation of mRNA degradation rates. Alternatively, com-
parative Dynamic Transcriptome Analysis (cDTA) (Sun
et al. 2012) (an updated version of Dynamic Transcriptome
Analysis [DTA]) (Miller et al. 2011) estimates rates of mRNA
degradation by determining the ratio of labeled to total
RNA using hybridization to a DNA microarray at a single
time point following addition of 4tU. However, cDTA re-
quires the manufacture of customized dual species DNA mi-
croarrays to normalize hybridization signals, and relies on a
single time-point after labeling, which may not accurately A variety of high-throughput methods have been intro-
duced with the goal of estimating in vivo rates of either
RNA synthesis or degradation. Genomic run on assays
(García-Martínez et al. © 2014 Neymotin et al.
This article, published in RNA, is available under a
Creative Commons License (Attribution 4.0 International), as described at
http://creativecommons.org/licenses/by/4.0/. METHOD METHOD ABSTRACT The abundance of a transcript is determined by its rate of synthesis and its rate of degradation; however, global methods for
quantifying RNA abundance cannot distinguish variation in these two processes. Here, we introduce RNA approach to
equilibrium sequencing (RATE-seq), which uses in vivo metabolic labeling of RNA and approach to equilibrium kinetics, to
determine absolute RNA degradation and synthesis rates. RATE-seq does not disturb cellular physiology, uses straightfor-
ward normalization with exogenous spike-ins, and can be readily adapted for studies in most organisms. We demonstrate
the use of RATE-seq to estimate genome-wide kinetic parameters for coding and noncoding transcripts in Saccharomyces
cerevisiae. Keywords: RATE-seq; RNA degradation; RNA synthesis; thiouracil; metabolic labeling Corresponding author: dgresham@nyu.edu
Article published online ahead of print. Article and publication date are at
http://www.rnajournal.org/cgi/doi/10.1261/rna.045104.114. Freely available
online through the RNA Open Access option. Determination of in vivo RNA kinetics using RATE-seq AMIN NEYMOTIN, RODONIKI ATHANASIADOU, and DAVID GRESHAM BENJAMIN NEYMOTIN, RODONIKI ATHANASIADOU, and DAVID GRESHAM BENJAMIN NEYMOTIN, RODONIKI ATHANASIADOU, and DAVID GRESHAM
Center for Genomics and Systems Biology, Department of Biology, New York University, New York, New York 10003, USA J
,
,
Center for Genomics and Systems Biology, Department of Biology, New York University, New York, New York 10003, USA Center for Genomics and Systems Biology, Department of Biology, New York University, New York, New York 10 Thiouracil labeling follows approach to equilibrium
kinetics The rate of change in RNA abundance (d[RNA]/dt) can
be modeled as a function of a constant rate of synthesis (k)
and a degradation rate proportional to RNA abundance
(α[RNA]) using the relationship d[RNA]/dt = k-α[RNA]. If
a labeled nucleotide is added to the culture the concentration
of labeled transcript will increase with time to an equilibrium
value at a rate solely determined by the transcript’s degrada-
tion rate constant (αRNA) and the cells’ division rate constant
(αgrowth) (i.e., α = αRNA + αgrowth). Approach to equilibrium
labeling, using radiolabeling, was developed over 40 yr ago
to estimate the rate of total mRNA turnover (Greenberg
1972) and was subsequently used to study individual tran-
scripts using transcript-specific probes (Harpold et al. 1981;
Kim and Warner 1983). To apply approach to equilibrium la-
beling on a genome-wide scale we developed a method using
4tU-labeling and RNA-seq (Fig. 1A). Our method relies on
the presence of an endogenous copy of uracil phosphoribosyl-
transferase (UPRT) in Saccharomyces cerevisiae (encoded by
FUR1), which converts 4tU into 4-thiouridine monophos-
phate allowing its incorporation in RNA. For the purpose
of normalizing RNA-seq libraries from different time points
following labeling, we added a constant quantity of three dif-
ferent in vitro-transcribed thiolated transcripts (Supplemen-
tal Table S1) to isolated RNA prior to fractionation. As Here, we report a general method for accurate measure-
ment of absolute RNA kinetic parameters in vivo. We use ap-
proach to equilibrium labeling (Greenberg 1972), which
minimizes exposure of cells to 4tU and is not affected by nu-
cleotide recycling. We undertook a series of rigorous controls
to optimize each step of the RATE-seq method. By using
strand-specific sequencing (Parkhomchuk et al. 2009) in
combination with ribosomal depletion, we measured rates
of decay for a variety of different types of RNA, including
noncoding RNA and snRNA. We developed a normalization
method using multiple spike-in RNAs that also enables iden-
tification and correction for technical artifacts. To account
for the nature of the data (i.e., overdispersed count data in
which the variance is greater than the mean) in model fitting
we used a weighted nonlinear regression to estimate param-
eters. Thiouracil labeling follows approach to equilibrium
kinetics We used RATE-seq to define the regulatory landscape
of steady-state transcript levels, defined as a function of the
underlying kinetic parameters genome-wide, and find that
many transcripts in budding yeast have similar steady-state
levels but differ greatly in their rates of production and
degradation. RATE-seq can be readily implemented in any
organism, making it a generally applicable method for char-
acterizing the steady-state in vivo kinetics of RNA with un-
precedented resolution. 1. RNA isolation
2. Addition of RNA spike-ins
3. HPDP biotinylation
4. Streptavidin pull-down
5. rRNA depletion
6. directional RNA-seq
A
time
RNA
4tU
td
Yeq
0
5
10
15
20
25
30
0
0.5e6
1.0e6
1.5e6
2.0e6
2.5e6
3.0e6
time (minutes)
signal (a.u.)
B
t1
t2
t3
t4
t5
t6
t1
t2
t3
t4
t5
t6
labeled RNA
unlabeled RNA
labeled RNA
t1
t2
t3
t4
t5
t6
FIGURE 1. RATE-seq enables in vivo measurement of RNA kinetics. (A) Overview of approach to equilibrium labeling and analysis using RATE-seq. The increase in labeled transcript with time Y(t) modeled using the relationship Y(t) = Yeq(1 −e−(aRNA+agrowth)(t−td)), where Yeq is the abundance of
labeled transcript at steady state, αRNA is the transcript’s degradation rate constant, αgrowth is the growth rate constant of the culture, t is the time after
addition of label, and td is a time delay between the addition of label and the time at which labeled transcripts can be detected. Red arrows indicate
points at which the RNA samples are recovered following addition of 4tU. (B) Incorporation of 4tU conforms to approach to equilbrium kinetics. An
equivalent quantity of biotinylated polyadenylated RNA from timepoints following addition of 4tU was bound to a membrane and visualized using
streptavidin alkaline phosphatase and chemifluorescence. Values are shown along with the model fit. B 1. RNA isolation
2. Addition of RNA spike-ins
3. HPDP biotinylation
4. Streptavidin pull-down
5. rRNA depletion
6. directional RNA-seq
A
time
RNA
4tU
td
Yeq
t1
t2
t3
t4
t5
t6
t1
t2
t3
t4
t5
t6
labeled RNA
unlabeled RNA
labeled RNA
t1
t2
t3
t4
t5
t6 0
5
10
15
20
25
30
0
0.5e6
1.0e6
1.5e6
2.0e6
2.5e6
3.0e6
time (minutes)
signal (a.u.)
B 5. rRNA depletion
6. directional RNA-seq FIGURE 1. RATE-seq enables in vivo measurement of RNA kinetics. (A) Overview of approach to equilibrium labeling and analysis using RATE-seq. RESULTS capture kinetic parameters. Indeed, the use of different indi-
vidual time points has a significant effect on the estimated
degradation rates for at least a subset of transcripts (Dölken
et al. 2008), which is likely the case for similar approaches us-
ing RNA-seq (e.g., Schwanhäusser et al. 2011). INTRODUCTION 2004) provide a means of estimating
mRNA synthesis rates; however, these methods require isola-
tion of nuclei or permeabilization of cells, which likely com-
promises the physiology of cells. Until recently, mRNA decay
rates have been estimated using transcriptional inhibition
(Wang et al. 2002; Grigull et al. 2004; Shalem et al. 2008) using
either temperature-sensitive alleles of RNA polymerase II or
chemical inhibition of transcript production. While these
methods succeed in inhibiting transcript synthesis, they typi- 1645 RNA 20:1645–1652; Published by Cold Spring Harbor Laboratory Press for the RNA Society Neymotin et al. Measurement of RNA degradation rates
transcriptome-wide deadenylated transcripts can persist in the cytoplasm or be
readenylated in some species (Wilt 1973), we used rRNA
depletion rather than poly(A) fractionation. The lack of
poly(A) selection step also enables the analysis of both coding
and noncoding transcripts. We performed RATE-seq using replicate yeast populations
growing in a defined rich medium during log phase. Fol-
lowing RNA-seq analysis, the relative counts (Supplemental
Tables S2, S3) of spike-ins are observed to decrease with
time and concomitantly, the proportion of counts mapping
to the transcriptome increases (Fig. 2A). We found that the
use of multiple spike-ins facilitated identification of technical
biases potentially introduced during library preparation
(Supplemental Fig. S8). The correlation of per transcript
counts between replicates at the same time point is high
(Spearman ρ = 0.98; Supplemental Fig. S9). To normalize
transcript counts (Supplemental Table S4) we first deter-
mined the ratio of counts for each transcript to each spike-
in, scaled each ratio, and then multiplied by the mean count
of all spike-ins across all experiments to preserve the scale of
the data (Materials and Methods). We studied the mean-var-
iance relationship at each time point and found that the data
are overdispersed (Supplemental Fig. S10). Therefore, to es-
timate the degradation rate constant for individual transcripts
we performed a nonlinear weighted regression using normal-
ized counts from the combined data set (Fig. 2B) (Materials
and Methods). Wedetermined confidence intervalsfor thees-
timated decay constant for each transcript using bootstrapped
values from each time-point (Materials and Methods). g
p
We first tested the efficiency of 4tU incorporation in S. cerevisiae and its physiological consequences. Consistent
with previous reports (Munchel et al. 2011) we find that yeast
cells take up 4tU provided in the growth medium and incor-
porate it into RNA (Supplemental Fig. S1). However, we find
that cells lacking a functional uridine monophosphate bio-
synthetic pathway (i.e., ura3−) cannot grow when supple-
mented with 4tU alone (Supplemental Fig. S2), suggesting
that highly thiolated transcripts are not tolerated by the cell. As we found comparable 4tU incorporation in a ura3−strain
and prototrophic strain (Supplemental Fig. S1) we performed
all subsequent experiments in a prototrophic strain. Over
the timescale and concentrations of 4tU used for RATE-seq
we detect no effect on cell growth (Supplemental Fig. S3),
although prolonged exposure and higher concentrations ap-
pear to have slight effects (Supplemental Fig. S4). Thiouracil labeling follows approach to equilibrium
kinetics The increase in labeled transcript with time Y(t) modeled using the relationship Y(t) = Yeq(1 −e−(aRNA+agrowth)(t−td)), where Yeq is the abundance of
labeled transcript at steady state, αRNA is the transcript’s degradation rate constant, αgrowth is the growth rate constant of the culture, t is the time after
addition of label, and td is a time delay between the addition of label and the time at which labeled transcripts can be detected. Red arrows indicate
points at which the RNA samples are recovered following addition of 4tU. (B) Incorporation of 4tU conforms to approach to equilbrium kinetics. An
equivalent quantity of biotinylated polyadenylated RNA from timepoints following addition of 4tU was bound to a membrane and visualized using
streptavidin alkaline phosphatase and chemifluorescence. Values are shown along with the model fit. 1646 RNA, Vol. 20, No. 10 RATE-seq Measurement of RNA degradation rates
transcriptome-wide We con-
firmed that the concentration of 4tU used for RATE-seq
does not affect global gene expression (Supplemental Fig. S5). Using a dot blot and colorimetric assay (Materials and
Methods), we find that the pool of newly synthesized
mRNA approaches equilibrium consistent with a model of
constant synthesis and exponential degradation (Fig. 1B). Consistent with expectation, the equilibrium value of labeled
RNA differs with different concentrations of 4tU, but the
kinetics of the approach to equilibrium is unaffected (Sup-
plemental Fig. S6). As with radiolabeling experiments in
mammalian cells (Greenberg 1972), the mRNA fraction ap-
proaches equilibrium faster than total RNA (Supplemental
Fig. S7), which reflects the relative stabilityof rRNA compared
with mRNA. Using RATE-seq we determined degradation rate con-
stants, and corresponding half-lives, with 95% confidence in-
tervals (CI) for 5308 mRNAs (Supplemental Table S5). Most
transcripts are rapidly degraded, with a mean and median
half-life of 15 and 10 min, respectively (Fig. 2C). Thus,
RATE-seq analysis estimates RNA half-lives that are shorter
than most previous global estimates (Wang et al. 2002;
Grigull et al. 2004; Miller et al. 2011; Munchel et al. 2011). 0.85
0.90
0.95
1.00
0.00
0.05
0.10
0.15
Proportionspike−in
0
5
10
15
20
25
30
time (minutes)
A
Proportiontranscriptome
0
20
40
60
80
100
0
500
1000
1500
time (minutes)
Counts
B
0
10
20
30
40
50
60
0
200
400
600
800
1000
1200
half−life (minutes)
Frequency
C
FIGURE 2. Global RNA kinetics determined using RATE-seq. (A) The relative fraction of reads mapping to the transcriptome increases with time,
whereas the fraction of reads mapping to spike-ins decreases. (B) Representative example of RATE-seq data for a rapidly degraded gene (CTK1, purple)
and a slowly degraded gene (GIM4, red). 95% CI for the estimated degradation rate constant are indicated by dashed lines. (C) The distribution of
half-lives for all coding transcripts with the mean (red line) and median (blue line) half-life indicated. 0.85
0.90
0.95
1.00
0.00
0.05
0.10
0.15
Proportionspike−in
0
5
10
15
20
25
30
time (minutes)
A
Proportiontranscriptome 0
20
40
60
80
100
0
500
1000
1500
time (minutes)
Counts
B 0
10
20
30
40
50
60
0
200
400
600
800
1000
1200
half−life (minutes)
Frequency
C A B C FIGURE 2. Global RNA kinetics determined using RATE-seq. The landscape of regulated transcript abundance At steady state, transcript abundance levels are constant (i.e.,
d[RNA]/dt = 0) and transcript synthesis and degradation are
related by the expression k = α[RNA]. Therefore, the rate of
transcript production can be estimated using the degradation
rate constant and the steady-state abundance of the tran-
script. As only a fraction of transcripts are labeled with 4tU
(Fig. 1A), RATE-seq does not quantify RNA abundance. Therefore, we used published estimates of absolute transcript
abundance from quantitative sequencing data (Lipson et al. 2009) to estimate rates of mRNA synthesis in steady-state
conditions (Supplemental Table S5). Our estimates of
mRNA synthesis rates are in good agreement with previous
estimates using Genomic-Run-On (GRO) assays (Supple-
mental Fig. S14; Pelechano et al. 2010) with a linear correla-
tion coefficient of r = 0.5. A source of discrepancy between
the two data sets may be that our study estimates the rate
of production of mature transcripts, whereas GRO estimates
nascent transcription rates. We compared mRNA half-lives estimated using RATE-seq
to previously reported estimates in Saccharomyces cerevisiae
(Supplemental Fig. S11). As noted in previous reports (Miller
et al. 2011; Munchel et al. 2011), the agreement among
mRNA half-lives using different methods is poor. Surpris-
ingly, RATE-seq estimates correlate poorly with those report-
ed using pulse-chase labeling with 4tU (Munchel et al. 2011). Our method has a number of differences that may account
for this including the absence of poly(A) selection, the use
of multiple spike-ins for normalization, and the use of un-
transformed data for nonlinear model fitting, which avoids
errors introduced by linear transformation of data. In addi-
tion, mathematical modeling suggests that nucleotide recy-
cling may slow the observed chase resulting in a systematic
underestimation of mRNA decay rates (Supplemental Fig. The combinatorial effect of variation in synthesis and deg-
radation rates defines the landscape of regulated transcript A
0
20
40
60
80
100
0
1000
2000
3000
4000
time (minutes)
scaled counts
snRNA
half-life (minutes)
B
transcriptome
0
20
40
60
80
0.05
0.10
0.15
0.20
0.0
0.2
0.4
0.6
0.8
1.0
degradation (min-1)
synthesis (molecules/min)
C
0.05
0.10
0.15
exponential decay constant (α) (min-1)
helicase
cell cycle
cytoskeleton
transcriptome
mitochondrion
ribosome
translation
FIGURE 3. Coordinated regulation of mRNA abundance levels. (A) Decay rates for sets of functionally related genes defined by GO term categories
are nonrandomly distributed. Measurement of RNA degradation rates
transcriptome-wide (A) The relative fraction of reads mapping to the transcriptome increases with time,
whereas the fraction of reads mapping to spike-ins decreases. (B) Representative example of RATE-seq data for a rapidly degraded gene (CTK1, purple)
and a slowly degraded gene (GIM4, red). 95% CI for the estimated degradation rate constant are indicated by dashed lines. (C) The distribution of
half-lives for all coding transcripts with the mean (red line) and median (blue line) half-life indicated. www.rnajournal.org
1647 www.rnajournal.org
1647 Neymotin et al. Using bootstrapped values we find that for the majority of
transcripts the estimated degradation rates have confidence
intervals of ±20% (Supplemental Table S5). A previous study
(Wang et al. 2002) showed that transcripts encoding func-
tionally related gene products have similar decay rates. We
find that genes within the same Gene Ontology (GO) terms
also have similar decay rates (Supplemental Table S6) al-
though the agreement between the estimated rates from the
two studies is poor. Functional categories representing the
most rapidly degraded transcripts include “Helicase activity”
and “Regulation of cell cycle” whereas categories representing
the most stable transcripts include “Cytoplasmic translation”
and “Ribosome” (Fig. 3A). S12). Our estimates are most similar to results obtained using
DTA (Miller et al. 2011), which may reflect the fact that both
methods isolate newly synthesized transcripts following label
addition. Importantly, consistent with both existing in vivo
labeling methods in budding yeast (Miller et al. 2011; Mun-
chel et al. 2011), we find that the half-lives for ribosomal pro-
tein-coding genes is greater than the median half-life of all
mRNAs (Supplemental Fig. S13). In contrast, in all studies
using transcriptional inhibition ribosomal protein-coding
transcripts are found to degrade as rapidly as the transcrip-
tome average, which may reflect a stress response to the pro-
found impact on cell physiology caused by these methods. In addition to variation in mRNA degradation rates we
find evidence for variation in rates of noncoding transcripts
including small nuclear RNA (snRNA) and long noncoding
RNAs (lncRNA) (Fig. 3B). The population of snRNAs appear
to be more stable than coding transcripts (Fig. 3B) and have
similar half-lives, suggesting that the post-synthesis fate of
snRNAs is coordinately regulated. Strains and growth conditions Experiments were performed using either FY4 (MATa) or FY3
(MATa ura3-52), which are isogenic to S288C. All RATE-seq anal-
yses were performed using the prototrophic strain FY4 in which a
single colony was inoculated into an overnight culture in synthetic
complete medium containing 500 µM uracil. The saturated over-
night culture was back-diluted 1:50 into fresh medium of the
same composition. Log phase cells were treated with 4tU to a final
concentration of 500 µM. Cells were collected at multiple time
points over the course of 100 min by vacuum filtration onto nitro-
cellulose filters and immediately frozen in liquid nitrogen. Dot blot analysis Forisolationofpoly(A)RNAfromtotalRNA,Oligod(T)25magnetic
beads (New England Biosciences) were used in combination with a
12-tube magnetic rack. Beads were washed once in a binding buff-
er/wash buffer (20 mM Tris-HCl at pH 7.5, 500 mM LiCl, 1 mM
EDTA) similar to manufacturer recommendations except that DTT
was left out of the buffer, as this would cleave the RNA conjugated
to biotin-HPDP. At least 40 µg of total RNA was added to 200 µL of
beads. Samples were washed in 1x binding buffer, then 1x low-salt
buffer, and eluted from beads in TE buffer following incubation for
3 min at 50°C. DISCUSSION The abundance of a transcript is determined by both its rate
of synthesis and its rate of degradation. To fully characterize
the regulation of mRNA levels these rates must be uncoupled. Moreover, studying transcripts under their native control
is critical as transcript stability may depend on cis-acting fac-
tors that associate with promoter regions (Bregman et al. 2011; Trcek et al. 2011). RATE-seq is an efficient and general means of estimating
transcriptome-wide absolute rates of RNA synthesis and deg-
radation in steady-state conditions. In contrast to existing
methods, it does not interfere with the cell’s physiology, pro-
vides enhanced accuracy, obviates the potential impact of nu-
cleotide recycling, and can be applied to a variety of types of
transcripts on a genome-wide basis. In principle, incorpora-
tion of 4tU is feasible in all organisms using either endoge-
nous or heterologous expression of UPRT (Cleary et al. 2005). Alternatively, 4sU can be used in organisms without
endogenous nucleotide salvage pathways (Dölken et al. 2008). Therefore, we expect that RATE-seq will be of great
utility for investigating the relationship between RNA synthe-
sis and degradation in a variety of genotypes and organisms. RNA biotinylation and streptavidin pull down For biotinylation reactions 100 µg of total RNA was added to a sol-
ution of 10 mM Tris-HCl (pH 7.4), and 1 mM EDTA. Biotin-HPDP
(1 mg/mL) was added to a final concentration of 2 µg for each 1 µg
of RNA (Supplemental Fig. S15A). In addition, the three spike-in
RNAs were pooled and 12 ng of the mixture added to the reaction
mixture containing 100 µg of RNA sample. The reaction was al-
lowed to proceed for 3 h in the dark, after which reactants were re-
moved using chloroform extraction. RNA pellets were precipitated
with 1 volume of isopropanol and 1/10 volume of 5 M NaCl. RNA pellets were washed once with 75% ethanol and resuspended
in RNase-free water. The biotinylated RNA was fractionated from unlabeled RNA us-
ing streptavidin magnetic beads (NEB) (Supplemental Fig. S15B). Pull downs were performed essentially as previously described
(Zeiner et al. 2008). Beads were washed four times and then tran-
scripts were cleaved from magnetic beads using β-mercaptoethanol
(5%). RNA was precipitated with 1 volume isopropanol, 1/10 vol-
ume NaCl, and 3 µg of glycogen (Supplemental Fig. S15C). The landscape of regulated transcript abundance (B) RATE-seq can be used to estimate the kinetics of snRNAs (e.g., snR48 [pink circles]) and lncRNAs (e.g., ICR1 [blue
triangles]). The boxplot (inset) displays the half-lives for snRNA and all other transcripts. (C) The global relationship between transcript synthesis and
degradation in S. cerevisiae. The abundance of each mRNA is categorized into quartiles (first quartile: 0.001–1.58 mRNAs/cell [cyan], second quartile:
1.6–2.8 mRNAs/cell [blue], third quartile: 2.8–4.8 mRNAs/cell [green], fourth quartile: 4.8–66 mRNAs/cell [red]). A
0.05
0.10
0.15
exponential decay constant (α) (min-1)
helicase
cell cycle
cytoskeleton
transcriptome
mitochondrion
ribosome
translation 0
20
40
60
80
100
0
1000
2000
3000
4000
time (minutes)
scaled counts
snRNA
half-life (minutes)
B
transcriptome
0
20
40
60
80 A 0.05
0.10
0.15
0.20
0.0
0.2
0.4
0.6
0.8
1.0
degradation (min-1)
synthesis (molecules/min)
C C B exponential decay constant (α) (min-1) FIGURE 3. Coordinated regulation of mRNA abundance levels. (A) Decay rates for sets of functionally related genes defined by GO term categories
are nonrandomly distributed. (B) RATE-seq can be used to estimate the kinetics of snRNAs (e.g., snR48 [pink circles]) and lncRNAs (e.g., ICR1 [blue
triangles]). The boxplot (inset) displays the half-lives for snRNA and all other transcripts. (C) The global relationship between transcript synthesis and
degradation in S. cerevisiae. The abundance of each mRNA is categorized into quartiles (first quartile: 0.001–1.58 mRNAs/cell [cyan], second quartile:
1.6–2.8 mRNAs/cell [blue], third quartile: 2.8–4.8 mRNAs/cell [green], fourth quartile: 4.8–66 mRNAs/cell [red]). 1648 RNA, Vol. 20, No. 10 RATE-seq abundance (Fig. 3C). Withinthis landscape it is clear thatclas-
ses of transcripts defined by rapid synthesis and degradation
have equivalent steady-state levels to classes of transcripts
that are comparatively slowly synthesized and degraded. Understanding the sources and consequences of these differ-
ent kinetics is central to understanding gene expression
regulation. EcoRI restriction and run off transcription performed as recom-
mended by the manufacturer with the addition of thiolated UTP:
UTP at a ratio of 2:1 in the reaction. Spike-in RNA was subsequently
treated with DNAse and purified. RNA extraction RNA was purified from cells using a hot acid phenol/chloroform ex-
traction. Briefly, 750 μL of lysis buffer (10 mM EDTA, 10 mM Tris,
0.5%SDS) was added to each sample and vortexed. An equal volume
of acid phenol was then added to the sample and vortexed. Samples
were incubated for 1 h at 65°C with occasional vortexing. Filters
were removed and samples were placed on ice for 10 min. After cen-
trifugation, the aqueous phase was transferred to Phase Lock Gel
(PLG) tubes and an equal volume of chloroform added. The aque-
ous phase was collected and RNA was precipitated using two vol-
umes of 95% ethanol and 0.1 volume of 3 M Sodium Acetate. RNA pellets were washed with 70% ethanol twice and dried at
room temperature for half an hour and resuspended in RNAse
free water. Depletion of ribosomal RNA Following fractionation of thiolated transcripts, 100 ng was depleted
of 18S and 25S ribosomal transcripts. Two rounds of ribosomal
depletion were performed using LNA probes provided in the
Ribominus kit (Invitrogen). RNA was then precipitated using 2 vol-
umes ethanol, 1/10 volume 3 M sodium acetate, and glycogen. Pellets were resuspended in 6 µL of RNAse free water. Cinjk = Ainjk · bnj 4) To return the data to the original scale we then multiplied the
normalized ratio for each gene by the average spike-in count
across all K time points from both replicates: Ninjk = Cinjk ·
j=2
j=1 Snjk
j=2
j=1 Kj
Library preparation for Illumina sequencing We excluded all data from a time-point if a spike-in was deviant in
its expected behavior. Thus, each time point has between two and
six values depending on whether the time point was replicated
and whether any data were removed. First strand synthesis of rRNA-depleted RNA was performed using
the Super Script III kit (Life Technologies) and random priming us-
ing random hexamers. Second-strand synthesis was performed with
dUTP in place of dTTP to enable strand-specific sequencing
(Parkhomchuk et al. 2009). Samples were end repaired, A-tailed,
and ligated to NEXTflex DNA Barcodes for multiplex sequencing. Adapter dimers were removed using AMPure beads (Agencourt). Samples were then treated with UNG and amplified using 10 cycles
of PCR prior to sequencing. Samples were sequenced using an
Illumina 2000 single-end 50-bp run. Sequence alignment 2. Yss = Yunlabeled + Ylabeled, Ilumina sequencing reads were first filtered for rRNA sequences
by aligning to the ribosomal DNA of the yeast genome using
Bowtie with default settings. All remaining reads were then aligned
to the rest of the yeast genome and the three spike-in sequences using
Bowtie2 (Langmead et al. 2009). After converting SAM files to BAM
files, reads were filtered based on quality scores of 20 or higher. The
resulting BAM files were then used to calculate total counts per tran-
script using the featureCounts function of the Subread package in R,
using the argument for strand specific counting. Each library had be-
tween 5 and 13 million reads mapping to non-rRNA transcripts. where Yunlabeled is the abundance of unlabeled transcript, Ylabeled is
the abundance of labeled transcript, Yss is the total abundance of a
transcript, and α is the transcript’s degradation rate constant. The
approach to equilibrium equation is then obtained by substitution
of equation 1 for the value of Yunlabeled in equation 2 and solving
for Ylabeled leading to Ylabeled = Yss(1 −e−at). Based on this equation, we modeled the abundance of labeled
transcript for each mRNA as Y(t) = Yeq(1 −e−(aRNA+agrowth)(t−td)),
where Y(t) is the amount of the labeled transcript at time t, Yeq is
the abundance of labeled transcript at steady state, αRNA is the tran-
script’s degradation rate constant, αgrowth is the growth rate constant
of the culture, t is the time after addition of label, and td is a time
delay between the addition of label and the time at which labeled
transcripts can be detected. Synthesis of polyadenylated thiolated spike-in RNAs To generate three RNA spike-ins with similar GC content to S. cerevisiae mRNAs, we cloned three different regions of the Bacillus
subtilis genome. The three spike-ins (spike-in700, spike-in900,
spike-in1200) have a GC content of 0.42 and lengths of 700, 900,
and 1200 bases, respectively. Three regions of the B. subtilis genome
were PCR amplified and cloned into the pSP64 poly(A) in vitro
transcription vector (Promega). Plasmids were linearized using For each sample, 200 ng of mRNA was blotted onto a Zeta-Probe
nylon membrane (BioRad) using a BioRad DotBlot. The RNA was
cross-linked using a UV cross-linker. The blot was blocked using
blocking buffer (PBS, 10% SDS, 1 mM EDTA) for 20 min. Samples
were then probed with Streptavidin Alkaline phosphatase in 1649 www.rnajournal.org Neymotin et al. 2) We then computed a scaling factor, β, for each spike-in by calcu-
lating the average ratio between each spike-in and a reference
spike-in across all K time points within a replicate j: blocking solution (1:1000). The membrane was washed in PBS atde-
creasing concentrations of SDS (10%, 1%, 0.1%) for 10 min each. Spots were visualized using ECF substrate (GE Healthcare), visual-
ized on a Typhoon FLA 9500, and analyzed using ImageQuant
software. blocking solution (1:1000). The membrane was washed in PBS atde-
creasing concentrations of SDS (10%, 1%, 0.1%) for 10 min each. Spots were visualized using ECF substrate (GE Healthcare), visual-
ized on a Typhoon FLA 9500, and analyzed using ImageQuant
software. bnj = (k=Kj
k=1 Snjk/Sn=1jk)
Kj 3) To normalize the data within a replicate, j we multiplied the ra-
tio, A for each gene by the scaling factor: Model fitting The approach to equilibrium method assumes transcript decay fol-
lows first order kinetics and transcript synthesis follows zeroth order
kinetics. This leads to the following two equations: 1. Yunlabeled = Ysse−at,
2. Yss = Yunlabeled + Ylabeled, 1. Yunlabeled = Ysse−at, 1. Yunlabeled = Ysse
,
2. Yss = Yunlabeled + Ylabeled, Received March 4, 2014; accepted July 7, 2014. ACKNOWLEDGMENTS We estimated the amount of 4tU labeling using a colorimetric Dot
Blot analysis of labeled RNA and a synthesized oligonucleotide con-
taining a 5′ biotin label. A standard curve was generated by diluting
known quantities of the labeled oligonucleotide and used to estimate
the number of labels in an RNA sample of known mass. Assuming
an average transcript length of 1200 nucleotides we estimate that ap-
proximately one out of 500 uracil is labeled after 35 min of labeling
under our conditions. We thank Christine Vogel, Daniel Tranchina, and members of the
Gresham lab for helpful comments. This work was supported by
the National Institutes of Health (GM107466 to D.G.); the National
Science Foundation (MCB-1244219 to D.G.); and a Dupont Young
Professor award (to D.G.). Author contributions: B.N. developed and optimized the RATE-
seq method and performed the experiments. R.A. developed the
RNA-seq library preparation protocol. B.N. performed computa-
tional analyses with help from R.A., and B.N. and D.G. conceived
of the method and wrote the paper with support from R.A. To test whether 4tU is preferentially incorporated, we performed
a DNA microarray analysis of 4tU labeled RNA compared with the
unfractionated sample (Supplemental Fig. S16). Consistent with
previous observations (Miller et al. 2009), there is a slight depend-
ency of label incorporation on length. Thus, RNA-seq analysis of
4tU labeled transcripts is expected to result in increased counts
for longer transcripts as a result of both increased labeling efficiency
and the larger target size of longer transcripts (Supplemental Fig. S17). This labeling bias affects the steady-state equilibrium value
for each transcript (Supplemental Fig. S18). However, this bias
does not affect the estimate of the decay rate as there is no relation-
ship between the counts of labeled transcripts at any time point and
our estimate of the decay rate constant (Supplemental Fig. S19). Accession codes We calculated 95% confidence intervals of the estimated deg-
radation rate constant by randomly sampling with replacement
the equivalent number of points from normalized data for each
mRNA 1000 times. When resampling fails to sample timepoints to-
ward the latter part of the curve we found that the nonlinear regres-
sion frequently failed to converge. Therefore, we used bootstrapped
values to estimate only α keeping Yeq the same for all iterations. Sequencing data are available through the Sequence Read Archive
under BioProject ID PRJNA236614. Gene enrichment analysis Gene enrichment analysis was performed as in (Gresham et al. 2010). Nonrandom distribution of decay rates for each GO SLIM
category as compared with the genes not in the category were iden-
tified using the Wilcoxon-Mann-Whitney test in R. Greenberg JR. 1972. High stability of messenger RNA in growing cul-
tured cells. Nature 240: 102–104. Gresham D, Boer V, Caudy A, Ziv N, Brandt NJ, Storey JD, Botstein D. 2010. System-level analysis of genes and functions affecting survival
during nutrient starvation in Saccharomyces cerevisiae. Genetics 187:
299–317. REFERENCES Bregman A, Avraham-Kelbert M, Barkai O, Duek L, Guterman A,
Choder M. 2011. Promoter elements regulate cytoplasmic mRNA
decay. Cell 147: 1473–1483. Cleary MD, Meiering CD, Jan E, Guymon R, Boothroyd JC. 2005. Biosynthetic labeling of RNA with uracil phosphoribosyltransferase
allows cell-specific microarray analysis of mRNA synthesis and de-
cay. Nat Biotechnol 23: 232–237. Dölken L, Ruzsics Z, Rädle B, Friedel CC, Zimmer R, Mages J,
Hoffmann R, Dickinson P, Forster T, Ghazal P, et al. 2008. High-res-
olution gene expression profiling for simultaneous kinetic parameter
analysis of RNA synthesis and decay. RNA 14: 1959–1972. Estimation of mRNA synthesis rates To estimate the synthesis rate for each transcript we assumed that
the rate of change in mRNA (dRNA/dt) at steady-state is equal to
zero and therefore used the relationship k = α[mRNA]. We also as-
sumed 60,000 mRNA/cell (Zenklusen et al. 2008). Confidence inter-
vals for mRNA synthesis rates were calculated using the 95% CI
values determined for αRNA. analysis of RNA synthesis and decay. RNA 14: 1959–1972 Elkon R, Zlotorynski E, Zeller KI, Agami R. 2010. Major role for mRNA
stability in shaping the kinetics of gene induction. BMC Genomics
11: 259. García-Martínez J, Aranda A, Pérez-Ortín JE. 2004. Genomic run-on
evaluates transcription rates for all yeast genes and identifies gene
regulatory mechanisms. Mol Cell 15: 303–313. Gasch AP, Spellman PT, Kao CM, Carmel-Harel O, Eisen MB, Storz G,
Botstein D, Brown PO. 2000. Genomic expression programs in the
response of yeast cells to environmental changes. Mol Biol Cell 11:
4241–4257. Data normalization We performed RATE-seq using two biological replicates, with time
points k = 3, 5, 7, 11, 13, and 25 min following label addition for rep-
licate 1, and time points k = 5, 7, 9, 13, 20, 25, 30, and 100 min fol-
lowing label addition for replicate 2. To normalize data within each
time series we used the following normalization scheme: To calculate the degradation rate constant for each transcript,
we performed nonlinear regression, estimating both α (the sum-
mation of αRNA and αgrowth) and Yeq. To account for biological
variation, we combined the data from both replicates to generate a
single parameter estimate for αRNA and Yeq. Because the variance
of the RATE-seq data increases with increasing time we used weight-
ed least squares regression with weights of 1/Y, which avoids un-
due influence of later time points on the model fit, using the gnls
function in the nlme package in R. To minimize the parameters
that we needed to estimate we set the time delay parameter, td equal
to 2 min, since labeled transcripts were pulled down as early as 3 min
after addition of 4tU. As the doubling time of the culture is 100 min,
αgrowth is equal to 0.0069. Transcript half-lives were calculated as
ln(2)/(α-0.0069). 1) We first computed a ratio, A, between the read count M, for each
gene i in replicate j at time point k and the read count S for each
spike-in n in replicate j at time point k: Ainjk = Mijk
Snjk where n = 1,2,3 for each of the three spike-ins and j = 1 or 2
depending on the replicate 1650 RNA, Vol. 20, No. 10 RATE-seq SUPPLEMENTAL MATERIAL The data and model fit for each gene can be visualized using the
available R script rateSeqFit.R using the function curve.generator(). Supplemental material is available for this article. R functions and packages Grigull J, Mnaimneh S, Pootoolal J, Robinson MD, Hughes TR. 2004. Genome-wide analysis of mRNA stability using transcription inhib-
itors and microarrays reveals posttranscriptional control of ribo-
some biogenesis factors. Mol Cell Biol 24: 5534–5547. All analyses were performed using R (R Core Team 2013) and sev-
eral open source packages. The functions featureCounts of the
Subread package and gnls of the nlme package were used for data
analysis of nonlinear regression. The following functions and pack-
ages, in addition to base functions in R and custom written func-
tions were used for presentation of figures: subplot of the
TeachingDemos package, axis.break of Plotrix, and heatscatter of LSD. Harpold MM, Wilson MC, Darnell JE. 1981. Chinese hamster polyade-
nylated messenger ribonucleic acid: relationship to non-polyadeny-
lated sequences and relative conservation during messenger
ribonucleic acid processing. Mol Cell Biol 1: 188–198. Kim CH, Warner JR. 1983. Messenger RNA for ribosomal proteins in
yeast. J Mol Biol 165: 79–89. 1651 www.rnajournal.org Neymotin et al. Langmead B, Trapnell C, Pop M, Salzberg SL. 2009. Ultrafast and mem-
ory-efficient alignment of short DNA sequences to the human ge-
nome. Genome Biol 10: R25. R Core Team. 2013. R: a language and environment for statistical
computing. R Foundation for Statistical Computing, Vienna,
Austria. http://www.R-project.org/. Lipson D, Raz T, Kieu A, Jones DR, Giladi E, Thayer E, Thompson JF,
Letovsky S, Milos P, Causey M. 2009. Quantification of the yeast tran-
i
b
i
l
l
l
i
N
Bi
h
l 27 Lipson D, Raz T, Kieu A, Jones DR, Giladi E, Thayer E, Thompson JF,
Letovsky S, Milos P, Causey M. 2009. Quantification of the yeast tran-
scriptome by single-molecule sequencing. Nat Biotechnol 27:
652–658. Schier AF. 2007. The maternal-zygotic transition: death and birth of
RNAs. Science 316: 406–407. scriptome by single-molecule sequencing. Nat Biotechnol 27:
652–658. Schwanhäusser B, Busse D, Li N, Dittmar G, Schuchhardt J, Wolf J,
Chen W, Selbach M. 2011. Global quantification of mammalian
gene expression control. Nature 473: 337–342. Miller MR, Robinson KJ, Cleary MD, Doe CQ. 2009. TU-tagging: cell
type–specific RNA isolation from intact complex tissues. Nat
Methods 6: 439–441. g
p
Shalem O, Dahan O, Levo M, Martinez MR, Furman I, Segal E, Pilpel Y. 2008. Transient transcriptional responses to stress are generated by
opposing effects of mRNA production and degradation. Mol Syst
Biol 4: 223. 1652
RNA, Vol. 20, No. 10 R functions and packages Miller C, Schwalb B, Maier K, Schulz D, Dümcke S, Zacher B, Mayer A,
Sydow J, Marcinowski L, Dölkin L, et al. 2011. Dynamic transcrip-
tome analysis measures rates of mRNA synthesis and decay in yeast. Mol Syst Biol 7: 458. Sun M, Schwalb B, Schulz D, Pirkl N, Etzold S, Larivière L, Maier KC,
Seizl M, Tresch A, Cramer P. 2012. Comparative dynamic transcrip-
tome analysis (cDTA) reveals mutual feedback between mRNA syn-
thesis and degradation. Genome Res 22: 1350–1359. y
Munchel SE, Shultzaberger RK, Takizawa N, Weis K. 2011. Dynamic
profiling of mRNA turnover reveals gene-specific and system-wide
regulation of mRNA decay. Mol Biol Cell 22: 2787–2795. y
thesis and degradation. Genome Res 22: 1350–1359. g
Trcek T, Larson DR, Moldón A, Query CC, Singer RH. 2011. Single-
molecule mRNA decay measurements reveal promoter- regulated
mRNA stability in yeast. Cell 147: 1484–1497. Nikolov EN, Dabeva MD. 1985. Re-utilization of pyrimidine nucleo-
tides during rat liver regeneration. Biochem J 228: 27–33. Wang Y, Liu CL, Storey JD, Tibshirani RJ, Herschlag D, Brown PO. 2002. Precision and functional specificity in mRNA decay. Proc
Natl Acad Sci 99: 5860–5865. Nonet M, Scafe C, Sexton J, Young R. 1987. Eucaryotic RNA polymerase
conditional mutant that rapidly ceases mRNA synthesis. Mol Cell
Biol 7: 1602–1611. Wilt FH. 1973. Polyadenylation of maternal RNA of sea urchin eggs after
fertilization. Proc Natl Acad Sci 70: 2345–2349. Parkhomchuk D, Borodina T, Amstislavskiy V, Banaru M, Hallen L,
Krobitsch S, Lehrach H, Soldatov A. 2009. Transcriptome analysis
by strand-specific sequencing of complementary DNA. Nucleic
Acids Res 37: e123. Zeiner GM, Cleary MD, Fouts AE, Meiring CD, Mocarski ES, Booth-
royd JC. 2008. RNA analysis by biosynthetic tagging using 4-thioura-
cil and uracil phosphoribosyltransferase. Methods Mol Biol 419:
135–146. Pelechano V, Chávez S, Pérez-Ortín JE. 2010. A complete set of nascent
transcription rates for yeast genes. PLoS One 5: e15442. Zenklusen D, Larson DR, Singer RH. 2008. Single-RNA counting reveals
alternative modes of gene expression in yeast. Nat Struct Mol Biol 15:
1263–1271. p
y
g
Puckett L, Chambers S, Darnell JE. 1975. Short-lived messenger RNA in
HeLa cells and its impact on the kinetics of accumulation of cyto-
plasmic polyadenylate. Proc Natl Acad Sci 72: 389–393. 1652
RNA, Vol. 20, No. 10
|
W4224308900.txt
|
https://link.springer.com/content/pdf/10.1007/s11615-022-00397-4.pdf
|
de
|
August, Vincent (2021): Technologisches Regieren. Der Aufstieg des Netzwerk-Denkens in der Krise der Moderne. Foucault, Luhmann und die Kybernetik
|
Politische Vierteljahresschrift
| 2,022
|
cc-by
| 1,012
|
Polit Vierteljahresschr (2022) 63:571–573
https://doi.org/10.1007/s11615-022-00397-4
REZENSION POLITISCHE THEORIE UND IDEENGESCHICHTE
August, Vincent (2021): Technologisches Regieren. Der
Aufstieg des Netzwerk-Denkens in der Krise der Moderne.
Foucault, Luhmann und die Kybernetik
Bielefeld: transcript. 480 Seiten. 38,00 C
Daniel Schulz
Angenommen: 30. März 2022 / Online publiziert: 25. April 2022
© Der/die Autor(en) 2022
Die vorliegende Arbeit von Vincent August rekonstruiert die Entstehung der Kybernetik in der zweiten Hälfte des zwanzigsten Jahrhunderts. Damit soll ein Diskurs
des Regierens sichtbar werden, der bislang, so die These, von Beschreibungskategorien wie Technokratie und Neoliberalismus nur unzureichend bezeichnet wurde.
Auch wenn sich die Untersuchung dafür zunächst in die Grenzbereiche der Politischen Theorie begeben muss, so liegt doch eine Pointe von August darin zu zeigen,
wie groß der Einfluss des kybernetischen Paradigmas auf die Frage des Regierens
im Allgemeinen und auf zwei der Großtheorien des vergangenen Jahrhunderts im
Besonderen gewesen ist. So erkennt er im Werk von Michel Foucault und Niklas
Luhmann zentrale Motive dieses theoriegeschichtlichen Umbruchs wieder.
Die als Dissertationsschrift bei Herfried Münkler und Hartmut Rosa entstandene umfangreiche Abhandlung siedelt dabei zwischen Politischer Theorie, Ideengeschichte und Sozialtheorie. August geht in drei großen Schritten vor: Im ersten Schritt zeichnet er zunächst die Renaissance des Souveränitätsdenkens in der
Zeit unmittelbar nach dem Zweiten Weltkrieg nach. Dieser Schritt dient ihm dazu, die nachfolgenden Krisen dieser Souveränitätsidee und der damit verbundenen
Vorstellungen von staatlicher Handlungsfähigkeit, Autonomie und Steuerung umso deutlicher hervorheben zu können. Hier greift August zu einer innovativen und
durchaus unerwarteten Gliederung in unterschiedliche Souveränitätsnarrative, die
sowohl klassisch-nationalstaatliche Ordnungsvorstellungen wie aber auch die postkoloniale Widerstandstheorie eines Frantz Fanon umfassen. Der zweite Schritt bildet
dann aber erst den eigentlichen Kern der Untersuchung. Hier arbeitet August den
Technokratiediskurs der Hochmoderne auf und deutet ihn als eine Reaktion auf die
beginnende Steuerungskrise des souveränen Nationalstaates. Der Clou von Augusts
Daniel Schulz ()
Freie Universität Berlin, Berlin, Deutschland
E-Mail: daniel.schulz@fu-berlin.de
K
572
D. Schulz
Argumentation liegt nun darin, die Entstehung der Kybernetik nicht etwa naheliegend der technokratischen Entwicklung zuzuschlagen. Die Technokratie wird von
ihm vielmehr als ein letztes Aufbäumen des modernen Autonomiegedankens interpretiert, der im Moment seiner Krise zu einer gesteigerten mechanisch-technischen
Zweck-Mittel-Rationalität greift, um der zunehmenden Komplexität gesellschaftlicher Steuerungsaufgaben gerecht zu werden. In diesem Krisenmoment der hochmodernen Steuerungsansprüche entwickelt sich nun die Kybernetik laut August als
eine radikale Alternative zum technokratischen Modell des Regierens. Die von ihm
sogenannte „technologische Gesellschaftskritik“ kritisiert die „veraltete Rationalität
der Moderne“ (S. 200), weil sie die neuen Herausforderungen von Differenz, Kontingenz und Autonomie der Subsysteme nicht angemessen darstellen kann. Damit
korrigiert August eine zeitdiagnostische Fehlwahrnehmung: Wenn die Entwicklung
der westlichen Demokratien in den vergangenen Jahrzehnten als Siegeszug des Neoliberalismus auf der Grundlage einer rationalen Handlungstheorie dargestellt wird,
so unterschlagen solche Deutungen nach August, dass es neben dieser Vorstellung
auch noch ein kybernetisch geprägtes technologisches Narrativ gegeben hat, in dem
Fragen der gesellschaftlichen Autonomisierung eine ungleich größere Rolle spielten. Im dritten Schritt widmet sich August schließlich der umfangreichen Diskussion
von Foucault und Luhmann und zeigt, an welchen Kategorien sich der Einfluss des
kybernetischen Denkens nachweisen lässt.
Nun ließen sich Einwände erheben, ob diese Entwicklungsskizze nicht zu großflächig erscheint, zumal dann, wenn man das neoliberale Modell – ob in der affirmativen Selbst- oder kritischen Fremdbeschreibung – ohnehin schon für einen
Scheinriesen gehalten hat. Die große Stärke der Arbeit besteht aber gerade darin,
dass August seine Abgrenzung der Diskurse immer wieder kenntnisreich einordnet
und präzise justiert, dabei auch gegenläufige Tendenzen und Positionen nicht unterschlägt. So wird der kybernetische Diskurs nicht einfach als die Negation der
staatlichen Steuerungstechniken oder des Neoliberalismus präsentiert, sondern als
ambivalente Transition von Government zur Governance eingeordnet.
Gerade weil die Rekonstruktion so filigran gearbeitet ist, fallen aber auch einige Leerstellen auf: So hätte im Diskurs der Technokratiekritik sicherlich Günther
Anders einen Platz verdient, auch Hans Blumenberg oder Lewis Mumford fehlen.
Hinzu kommt, dass die Abkehr vom handlungsfähigen Subjekt nicht nur in der
Netzwerktheorie und der Kybernetik stattfand. Auch die bis heute einflussreiche
Verhaltenslehre des Behaviorismus, der Sozialpsychologie und der kognitionswissenschaftlich basierten Ökonomie haben die politische Ordnungsvorstellung vom
handlungsfähigen Individual- und Kollektivsubjekt nachhaltig unterminiert. Zudem
hätten Verbindungen zum genuin politikwissenschaftlichen Steuerungsdenken beispielsweise eines Harold Lasswell nahegelegen. Auch ließe sich fragen, ob Regierung, Regulierung und Steuerung nicht trennschärfer unterschieden werden müssten.
Dies ändert jedoch nichts an der Tatsache, dass August ein bislang vernachlässigtes Forschungsgebiet für die sozialwissenschaftliche Diskussion auf beeindruckende
Weise neu erschlossen hat. Die Komplexität seiner Rekonstruktion ermöglicht zudem eine ganze Reihe von produktiven Anschlussfragen. So ließe sich die theoriegeschichtliche Überlegung mit der Frage erweitern, ob das Zeitalter der Digitalisierung
die Renaissance eines technischen Steuerungs- und Verfügbarkeitsdenkens darstellt,
K
August, Vincent (2021): Technologisches Regieren. Der Aufstieg des Netzwerk-Denkens in der...
573
oder ob die kybernetische Diagnose sich gegenseitig abschließender, für staatliche
oder politische Zugriffe unverfügbare autonome Systeme die „Grenzen der Verfügbarkeit“ (S. 163) aufzeigt. Auch wäre von Interesse, inwieweit man in den von
August gezeichneten Entwicklungen auch ein normatives Potenzial erblicken kann:
Liegt in der Ausdifferenzierung gesellschaftlicher Subsysteme nicht selbst ein Autonomiegarant gegen die übergreifenden Steuerungsansprüche nicht mehr staatlicher,
sondern zunehmend privater Akteure? Läge dann die Bedrohung demokratischer Gesellschaftsordnungen nicht zuerst in einer diffusen Kolonialisierung der Lebenswelt
(Habermas), sondern in den potenziellen Entdifferenzierungseffekten digitaler Technologien? Die Arbeit von August macht einen wichtigen Schritt in die Richtung einer politischen Theorie des digitalen Zeitalters. Sie ist nicht nur ideengeschichtliche
Rekonstruktion, sondern auch kritische Zeitdiagnose der Transformation politischer
Möglichkeitsräume.
Funding Open Access funding enabled and organized by Projekt DEAL.
Open Access Dieser Artikel wird unter der Creative Commons Namensnennung 4.0 International Lizenz veröffentlicht, welche die Nutzung, Vervielfältigung, Bearbeitung, Verbreitung und Wiedergabe in
jeglichem Medium und Format erlaubt, sofern Sie den/die ursprünglichen Autor(en) und die Quelle ordnungsgemäß nennen, einen Link zur Creative Commons Lizenz beifügen und angeben, ob Änderungen
vorgenommen wurden.
Die in diesem Artikel enthaltenen Bilder und sonstiges Drittmaterial unterliegen ebenfalls der genannten
Creative Commons Lizenz, sofern sich aus der Abbildungslegende nichts anderes ergibt. Sofern das betreffende Material nicht unter der genannten Creative Commons Lizenz steht und die betreffende Handlung
nicht nach gesetzlichen Vorschriften erlaubt ist, ist für die oben aufgeführten Weiterverwendungen des
Materials die Einwilligung des jeweiligen Rechteinhabers einzuholen.
Weitere Details zur Lizenz entnehmen Sie bitte der Lizenzinformation auf http://creativecommons.org/
licenses/by/4.0/deed.de.
K
|
|
https://openalex.org/W4302385959
|
https://zenodo.org/records/6799719/files/Keletn%C3%A9met%20f%C3%B6ldekre%20vad%C3%A1sznak%20a%20nagybefektet%C5%91k.pdf
|
Hungarian
| null |
Keletnémet földekre vadásznak a nagybefektetők
|
Zenodo (CERN European Organization for Nuclear Research)
| 2,015
|
cc-by
| 1,874
|
émet földekre vadásznak a nagybefektetők Kategória: 2015. június
Írta: Rachel Knaebel Kategória: 2015. június
Írta: Rachel Knaebel A porosz nemesi nagybirtok után az erőszakos kollektivizálás és
az
óriás
termelőszövetkezetek
működtetése,
majd
a
posztszocialista mezőgazdasági átmenet után a nagybefektetők
kártékony nyomulása… Dióhéjban ennyi a keletnémet agrárium
története. Ma a keletnémet mezőgazdaság iránt újra felébredt az
érdeklődés, de gyakran az ágazattól idegen és külföldi
nagybefektetők vásárolnak fel komplett gazdaságokat − és ezzel
együtt több ezer hektár termőföldet. Ennek a helyzetnek paradox
módon kedvez a kommunista hatalomtól örökölt birtokszerkezet. Stefan Palme egy bio-gabonatermelő üzemet vezet Berlintől nyolcvan kilométerre. Az ötvenes
éveiben járó férfi a képernyőn egy mezőgazdasági földeket megjelenítő színes térképet mutat. A birtok
központi épületét a XVII. századi junkerek, a régi porosz nemesi nagybirtokosok építették. Ma ezen a
birtokon ezerszáz hektáron termelnek búzát, rozst, tönkölyt, árpát és zabot. A férfi a térképen színes
sokszögeket, parcellákat mutat: „a vörös színűek Brandenburg tartományé, a rózsaszínűek az
egyházé” − magyarázza. Azután a legnagyobb felületekre mutat: „Ezek a földek itt a Steinhoffé, azok
pedig ott a Thomas Philippsé.” A Steinhoff egy nagy német bútorgyártó cég, része egy nemzetközi holdingnak, amelynek
székhelye Dél-Afrikában van. A Thomas Philipps pedig egy outlet-üzletlánc. Ilyen versenytársakkal
szemben − ha földet akar vásárolni vagy bérelni − még az olyan jelentős birtokosoknak sincs komoly
súlya, mint Palme úr. „E pillanatban minden földdarabért harc folyik. Ez valóságos vadnyugat!”−
panaszkodik a bajor férfi, aki 1996 óta telepedett meg a régióban. Kevéssel az 1989-es újraegyesítés után a keletnémet mezőgazdaságba gazdálkodói múlt
nélküli, de pénzes befektetők érkeztek. Egy hulladékgazdálkodással foglalkozó ipari birodalom
(Remondis) tulajdonosa így vásárolt fel 1994-ben többet is a négyszázhatvanöt állami gazdaságból,
amelyek a Német Demokratikus Köztársaság (NDK) idején közvetlen állami kezelésben voltak. Ezek
az „össznépi tulajdonban levő” farmok a megművelt területeknek kevesebb, mint egytizedét tették ki. Az NDK mezőgazdasága inkább a mezőgazdasági termelőszövetkezeteken alapult, ahol a föld és a
gépek közös tulajdonban voltak. 1945-ben a szovjetek kisajátították és államosították a száz hektárnál
nagyobb birtokokat, 1960-ra befejeződött erőszakos kollektivizálás is, és úgy háromezer-nyolcszáz
szövetkezet alakult meg. Ezek több mint háromnegyede 1990 után új jogi formában, de fennmaradt és
folytatta tevékenységét[1]. Ezek állnak most a figyelem középpontjában… Keletnémet földekre vadásznak a nagybefektetők Keletnémet földekre vadásznak a nagybefektetők 2022. 07. 05. 20:56 Új befektetők méregetik az ország keleti részét https://www.magyardiplo.hu/archivum/2015-januar/195-2015-junius/1891-keletnemet-foldekre-vadasznak-a-nagybefektetok.html?tmpl=componen… um/2015-januar/195-2015-junius/1891-keletnemet-foldekre-vadasznak-a-nagybefektetok.html?tmpl=componen…
1/ 1/5 Keletnémet földekre vadásznak a nagybefektetők 2022. 07. 05. 20:56 A földek felvásárlásának módja és mértéke 2007-től kezdve, a gabonaárak emelkedése és a Wall
Street összeomlása óta, megváltozott. Wolfgang Krüger, a legnagyobb német mezőgazdasági
termelőket képviselő csoport, a Deutscher Bauernverband (DBV) osztályvezetője szerint: „A pénzügyi
válság új szereplőket vonzott ide. Azt mondják, hogy a mezőgazdasági területekbe való beruházás
biztos üzlet, még ha a hozama mérsékelt is”. A nyugatnémet befektetők között van a Steinhoff
bútorgyártó, és a Lindhorst is. (Utóbbi egy ingatlanokra és öregotthonokra szakosodott cégcsoport.) A
legnagyobb mezőgazdasági nagybefektető a KTG Agrar, amely 2007-ben lépett a tőzsdére. A
társasságnak ma Németország keleti részén harmincötezer hektár föld és harminc gazdaság van a
birtokában. Ez a szám folyamatosan növekszik. A KTG más befektetők nevében működik
Romániában és Litvániában, Oroszországban pedig egy német húsipari vállat számára termel
takarmányt. Ezek a társaságok csak a leggazdaságosabb termékek iránt érdeklődnek: gabonát, repcét
és biogázt termelnek, amelyeket szabályozott árak szubvencionálnak. A Rajnán túl ezeket a cégeket erősen bírálják, így roppant diszkréten viselkednek. A Steinhoff
és a Lindhorst egyáltalán nem nyilatkozik a sajtónak, a KTG pedig a public relation ügynökségéhez
irányítja az érdeklődőket. A közepes méretű Agro Energy beszédesebbnek látszik. A 2008-ban
alapított hamburgi cég úgy mutatja be részvényeseinek a keletnémet mezőgazdasági befektetéseket,
mint amelyek különösen ellenállnak az inflációs kockázatoknak, egyúttal hasznot is hajtanak. Az első
34 millió eurós alapgyűjtésnek köszönhetően a társaság megvásárolt két, összesen több mint négyezer
hektáros gazdaságot, amelyet 2011-ben átlagosan 13,5 százalékos haszonnal adott el. Az Agro Energy
most megpróbál összegyűjteni 120-150 millió euró tőkét, hogy további húszezer hektár földet
szerezzen. Eközben egész sor új befektető méregeti az ország keleti részét: egy logisztikai társaság
vezetősége, egy szemüvegeket áruló kiskereskedelmi lánc tulajdonosa, egy pénzügyi vállalkozás
korábbi menedzsere, egy volt nyomdaigazgató, de még az a cukortermelő is. Utóbbinak többezer
hektárnyi földje van Szászországban, Moldáviában, Lengyelországban és Chilében is. „A
nagyfelvásárlók nem közvetlenül a földet vásárolják meg, hanem vállalkozásokat vesznek, mellyel
ellenőrzésük alá vonhatják a földterületeket” − magyarázza M. Andreas Tietz, egy e tárgyban
megjelent munka társszerzője[2]. A német törvény ugyanis kizárólag mezőgazdasági termelőknek
engedi meg a földek eladását, ugyanakkor nem tiltja senkinek, hogy működő gazdaságokat
vásároljanak. A befektetők felismerték, hogy a gazdaságok megvétele a földek megszerzését is jelenti. A KTG és mások így szereztek óriási farmokat, melyekben a korábbi gazdákat egyszerű
alkalmazottként foglalkoztatják. A keleti tartományokban a földek háromnegyed részét haszonbérleti formában művelik[3]. https://www.magyardiplo.hu/archivum/2015-januar/195-2015-junius/1891-keletnemet-foldekre-vadasznak-a-nagybefektetok.html?tmpl=componen…
2/5 Új befektetők méregetik az ország keleti részét A
tulajdonosok többnyire magánszemélyek, korábbi szövetkezeti tagok, vagy olyan emigránsok, akik a
rezsim bukása után kárpótlásként jutottak parcellákhoz. Németország keleti régióiban a legnagyobb
földbirtokosok egyike az egyház. Az NDK idején ugyanis a hatalom formálisan nem vitatta el az
egyház földtulajdonát, a földhivatali nyilvántartásban a tulajdonuk megmaradt, ám a papság nem
művelhette, bérbe sem adhatta a földjeit. Az egyház jelenleg sem adja el a földjeit. A keletnémet mezőgazdasági befektetések iránti szenvedély több okra vezethető vissza. Egyrészt a földárakra, (2007-es adat szerint negyede a nyugat-németországi áraknak), másrészt az 2/5 chivum/2015-januar/195-2015-junius/1891-keletnemet-foldekre-vadasznak-a-nagybefektetok.html?tmpl=componen… Keletnémet földekre vadásznak a nagybefektetők 2022. 07. 05. 20:56 egykori NDK birtokszerkezetére. Keleten az átlagos birtoknagyság több mint kétszázharminc hektár,
jóval nagyobb mint Nyugaton, (átlagosan negyvenkilenc hektár) vagy mint Franciaországban (ötvenöt
hektár). Sok közülük eléri vagy meghaladja az ezer hektárt, főleg azok, amelyek közvetlen utódai a
korábbi mezőgazdasági termelőszövetkezeteknek. „A kollektivizálás idején a földeket egyesítették,
hogy nagy egységek jöjjenek létre” - magyarázza Arnd Bauerkämper történész, a Berlini Szabad
Egyetem professzora. „A német helyzet igencsak sajátos: az egyesítés egyúttal az Európai Unióba való
bekerülést is jelentette. Ez pedig kedvezett az egykori NDK birtokszerkezet fennmaradásának”. Furcsa történelmi helyzet, de a valamikori kollektivizálás most a nagy magántársaságok kezére
játszik. Wolfgang Krüger szerint ezek a nagybefektetők összesen talán százezer hektárt képviselnek. „Ez
kevesebb, mint 2 százaléka a keleti régiók öt és fél millió hektárnyi mezőgazdasági területének.”-
állítja Krüger úr, aki maga is mezőgazdasági szövetkezeti tagok gyermeke.A tulajdonosi koncentráció
azonban felgyorsult az utóbbi években, és bizonyos régiókban eléri a 10-25 százalékot is − legalábbis a
jó minőségű földek esetében. „Ez maga a neofeudalizmus” − állítja Helmut Klüter, a Greifswaldi
Egyetem régióföldrajz professzora. „A nemesi birtokrendszer idején a legnagyobb földek területe
ritkán érte el a négyszáz hektárt. Nem ezreket, mint manapság. És ezeket a gigászi vállalkozásokat
támogatja az Unió közös agrárpolitikája!” A KTG Agrar például saját bevallása szerint évente 6 millió
euró közösségi támogatáshoz jut. Ugyanez alatt az idő alatt Mecklenburg-Nyugat Pomeránia,
Szászország, Brandenburg német termelői a földárak robbanásától szenvednek. 2007 óta a keleti
régiókban kétszeresére, 1990 óta pedig a háromszorosára nőttek a földárak[4]. „A Lindhorst innen délkeletre telepedett meg, a KTG a szomszéd faluban vett egy gazdaságot”
− mondja Holger Lampe. Az 56 éves férfi egy mezőgazdasági szövetkezetet irányít a lengyel határ
közelében. Ő és a szervezet tizenkét tagja kétszáz tehenet tartanak ezernégyszáz hektáron, amelynek
több mint a fele bérelt föld. https://www.magyardiplo.hu/archivum/2015-januar/195-2015-junius/1891-keletnemet-foldekre-vadasznak-a-nagybefekt https://www.magyardiplo.hu/archivum/2015-januar/195-2015-junius/1891-keletnemet-foldekre-vadasznak-a-nagybefektetok.html?tmpl=componen…
3/5 Új befektetők méregetik az ország keleti részét „Ez itt valaha egy junker nagybirtoka volt” – mutatja a gazda, aki 1987 óta
irányítja a szövetkezetet. Mint fiatal agrármérnök érkezett ide, átélte a Fal leomlását, az egyesítést, a
piacgazdaságba való átmenetet, majd belemártózott a közös agrárpolitika „hideg vizes medencéjébe”,
és azóta sem találja a vezérfonalat. Ma az új befektetők érkezésével kell szembesülnie, és az
elérhetetlen földárakkal: „Az 1990-es években 1000 euróért vettünk egy hektárt. Ma ez 10-12 ezer
euróba kerülne, ami nekünk túl drága.” A húsz éve olcsó áron szerzett földeket a BVVG-től (Bodenverwertungs- und -verwaltungs
GmbH[5]), a gazdálkodásért és működtetéséért felelős köztestülettől vették meg. A társaság feladata
1992 óta a keletnémet állam tulajdonát képező négymillió hektár mezőgazdasági föld és erdő
privatizálása, amelyből több mint hétszázezer hektárt adott el nem kevesebb mint 6 milliárd euróért. És ez még nem minden… „A BVVG maradt Keleten a legnagyobb földtulajdonos, de az általa alkalmazott árak ma már
nem védhetők” − fogalmaz kritikusan Udo Folgart, egy mezőgazdasági csoportosulás elnöke, a
brandenburgi tartományi parlament választott képviselője. Folgart úr maga is egy keleti gazdaság
igazgatója az 1980-as évek óta. Hosszú évekig a BVVG baráti áron adott el földeket olyan cégeknek
mint az övé, vagyis a megmaradt keletnémet szövetkezeteknek. Ez a stratégia azonban 2007-ben
megváltozott: a földeket pályázatokon értékesítik, ahol a legmagasabb árakat ajánló fél nyer, ez pedig 3/5 chivum/2015-januar/195-2015-junius/1891-keletnemet-foldekre-vadasznak-a-nagybefektetok.html?tmpl=componen… Keletnémet földekre vadásznak a nagybefektetők 2022. 07. 05. 20:56 felhajtja az árakat és növeli az inflációt. A BVVG tagadja, de a saját dokumentumai is alátámasztják,
hogy a parcellákat jóval az átlagos árak felett adja el. felhajtja az árakat és növeli az inflációt. A BVVG tagadja, de a saját dokumentumai is alátámasztják,
hogy a parcellákat jóval az átlagos árak felett adja el. „Evidens, hogy a BVVG emelni fogja az árakat” − állítja Ralf Behring, aki egy százhektáros
családi farm tulajdonosa Brandenburgban. Ő és családja biotermelésben érdekelt, gabonát és almát
termel, illetve birkákat nevel, és a fővárosba szállítja eladásra. A férfi Németország délnyugati részén
nőtt fel; 1992-ben, mindössze 29 évesen költözött ide, és az újraegyesítés következtében kapta vissza
nagyapja korábbi családi gazdaságát. A nagyapa 1958-ban a szocialista hatalom elől menekült el. Ma
Behring úr került konfliktusba a hatalommal, a földeket kezelő állami társasággal, mert földjeinek egy
része a BVVG tulajdonában van, amely minél gyorsabban, és minél magasabb áron akarja azokat
eladni. „Ha a legmagasabb árat kínálóknak adják el, nem tudom megvásárolni a földeket −
panaszkodik a gazda. „Tudom, hogy el fogom veszíteni a földjeimet. Új befektetők méregetik az ország keleti részét Utána már csak a hétvégi farm
maradna meg nekünk, amit hétvégére bérbe adhatunk a városból érkező turistáknak.” A házaspár kis
családi gazdasága tehát a puszta túlélésért küzd. Till
Backhaus,
Mecklenburg-Nyugat-Pomeránia
tartományának
szociáldemokrata
mezőgazdasági minisztere egy egyszerű megoldást kínál a jelenlegi helyzetre: állítsák le a privatizációt
teljesen. „Meg akarjuk szerezni mérsékelt áron a tartomány megmaradt ötvenezer hektárnyi területét,
ami még a BVVG kezében van. Utána kiadjuk fiatal gazdáknak például biofarmok létesítésére, de
mindenképpen olyan gazdálkodás céljára, ami munkalehetőségeket teremt és értéket termel.”
Backhaus maga is szövetkezeti elnök volt egykor. Ötletének jelenleg nagyon kicsi esélye van a
megvalósításra. Fordította: Hrabák András [1] Michel Streith : Retour vers le futur. Les premières années de la transition agricole post-socialiste
dans l’ex-RDA [Vissza a jövőbe. A posztszocialista mezőgazdasági átmenet első évei az egykori NDK-
ban], Economie rurale, n° 325-326, Paris, 2011. [2] Bernhard Forstner, Andreas Tietz, Klaus Klare, Werner Kleinhanss és Peter Weingarten:
Aktivitäten von nichtlandwirtschaftlichen und uberregional ausgerichteten Investoren auf dem
landwirtschaftlichen
Bodenmarkt
in
Deutschland,
Sonderheft,
n°352,
Thünen-Institut,
Braunschweig, 2011. [3] Egész Németországban, csak a mezőgazdasági földek 38 százaléka van mezőgazdászok
tulajdonában. [3] Egész Németországban, csak a mezőgazdasági földek 38 százaléka van mezőgazdászok
tulajdonában. [3] Egész Németországban, csak a mezőgazdasági földek 38 százaléka van mezőgazdászok
tulajdonában. [4] 2013-ban, egy hektár termőföld közel kétszer drágább volt az egykori NDK-ban, mint
Franciaországban (átlagosan 10 500 euro vs. 5 750 euro). [4] 2013-ban, egy hektár termőföld közel kétszer drágább volt az egykori NDK-ban, mint
Franciaországban (átlagosan 10 500 euro vs. 5 750 euro). [5] A BVVG lépett a Treuhand helyére, amelyet 1990-ben hoztak létre az egykori NDK vállalatainak
privatizálására és 1994-ben szüntettek meg. [5] A BVVG lépett a Treuhand helyére, amelyet 1990-ben hoztak létre az egykori NDK vállalatainak
privatizálására és 1994-ben szüntettek meg. https://www.magyardiplo.hu/archivum/2015-januar/195-2015-junius/1891-keletnemet-foldekre-vadasznak-a-nagybefektetok.html?tmpl=componen… neoliberalizmus 4/5 Keletnémet földekre vadásznak a nagybefektetők Keletnémet földekre vadásznak a nagybefektetők 2022. 07. 05. 20:56 https://www.magyardiplo.hu/archivum/2015-januar/195-2015-junius/1891-keletnemet-foldekre-vadasznak-a-nagybefekt u/archivum/2015-januar/195-2015-junius/1891-keletnemet-foldekre-vadasznak-a-nagybefektetok.html?tmpl=componen… 5/5
|
https://openalex.org/W2150176294
|
https://eprints.bbk.ac.uk/id/eprint/27050/1/27050.pdf
|
English
| null |
UK Real‐Time Macro Data Characteristics
|
The economic journal/Economic journal
| 2,006
|
public-domain
| 8,450
|
▪ Birkbeck, University of London ▪ Malet Street ▪ London ▪ WC1E 7HX ▪ BIROn - Birkbeck Institutional Research Online Garratt,
Anthony and Vahey,
S.P. (2004) UK real-time macro data
characteristics. Working Paper. Birkbeck, University of London, London,
UK. Downloaded from: https://eprints.bbk.ac.uk/id/eprint/27050/ ∗We thank Alex Brazier, Dean Croushore, Colin Ellis, George Kapetanios, Simon van Nor-
den, Stephen Machin, Tony Yates and two anonymous referees for helpful comments.
We are
grateful to seminar participants at the University of Cambridge and the Reserve Bank of New
Zealand. Kateryna Rakowsky and Mutita Akusuwan provided excellent research assistance. Finan-
cial support from the Department of Applied Economics and from the ESRC (Research Grant No.
L38251021) is acknowledged gratefully. Shaun Vahey visited the University of British Columbia
during the period in which the first draft was completed. The views expressed in this paper are
those of the authors and do not necessarily represent those of the Reserve Bank of New Zealand. UK Real-time Macro Data Characteristics∗ Anthony Garratt
Birkbeck College Shaun P Vahey
Reserve Bank of New Zealand December 21, 2004 Birkbeck Working Papers in Economics & Finan
School of Economics, Mathematics and Statistics
BWPEF 0502
UK Real-Time Macro Data
Characteristics
Anthony Garratt
Shaun P Vahey
November 2004
▪ Birkbeck, University of London ▪ Malet Street ▪ London ▪ WC1E 7HX ▪ Birkbeck Working Papers i Anthony Garratt
Shaun P Vahey Bi ▪ Birkbeck, University of London ▪ Malet Street ▪ London ▪ WC1E 7HX ▪ Keywords: real-time data, structural breaks, probability event forecasts
JEL Classification: C22, C82, E00 1
Introduction In this paper, we characterise the revisions to a variety of commonly-used UK macro-
economic indicators. We find that the preliminary measurements of most real-side
macro indicators are downwards biased predictors of subsequent measurements (at
the sample means). Structural breaks affect the relationships between early and
later measurements for many variables. Previous studies, including (among others) Symons (2001), Castle and Ellis
(2002) and Mitchell (2004) have noted the predictability property for the expendi-
ture measure of output and its components. These (combined) studies characterise
a subset of the indicators considered in this paper and provide no formal analyses of
structural breaks. We use the Bai and Perron (2003a and 2003b) test for multiple
breaks of unknown timing to examine the time variation in predictability. Some of the UK indicators characterised in this study are drawn from two existing
real-time data sets, Castle and Ellis (2002) and Egginton, Pick and Vahey (2002). Only Castle and Ellis (2002) characterise the revisions processes in detail (for the
expenditure measure of output and its components). In addition, we analyse the
real-time quarterly monetary aggregates, nominal GDP and price deflator variables
neglected in the existing databases. The preliminary measurements of UK monetary
aggregates are largely unbiased. In contrast, initial nominal GDP and GDP price
deflator measurements typically understate final measurements. The revisions to
these nominal variables rarely exhibit structural breaks. The untypical behaviour of
monetary aggregate revisions reflects the very different collection processes for these
series. Macro models often perform better with revised data than with preliminary mea-
surements. Real-time data sets allow researchers to condition their ex post model
analyses on the information set actually available to forecasters and policymakers
in real time. But if researchers ignore the predictability in initial measurements,
real-time model performance can be misjudged. We illustrate this with a specific
forecasting example. We use a vector autoregression (VAR) in UK real output
growth and inflation to forecast the (one-step ahead) probability of above trend
growth–sometimes referred to as the likelihood of “positive momentum”. Ignoring
the predictability in initial measurements understates the event probability consid-
erably for our 1990Q1 to 1999Q2 evaluation period. The remainder of the paper is organised as follows. In section 2, we discuss the
sources of UK real-time data. We describe our methodology for characterising UK
real-time data in section 3 and report the main results in section 4. 1US
data
can
be
downloaded
from
the
Philadelphia
Federal
Reserve
Bank
http://www.phil.frb.org/econ/forecast. Croushore and Stark (2001) describe data set construction. 1
Introduction We analyse our
illustrative probability forecasting VAR exercise in section 5. Section 6 concludes. Abstract We characterise the relationships between preliminary and subsequent mea-
surements for 16 commonly-used UK macroeconomic indicators drawn from
two existing real-time data sets and a new nominal variable database. Most
preliminary measurements are biased predictors of subsequent measurements,
with some revision series affected by multiple structural breaks. To illustrate
how these findings facilitate real-time forecasting, we use a vector autoregresion
to generate real-time one-step-ahead probability event forecasts for 1990Q1 to
1999Q2. Ignoring the predictability in initial measurements understates con-
siderably the probability of above trend output growth. 1 2
Data sources Two on-line real-time UK data sources have appeared in the last couple of years:
Castle and Ellis (2002) and Egginton, Pick and Vahey (2002).1 Both studies adopt 2 the standard terminology used in the more recent literature to describe the data
(see, for example, Diebold and Rudebusch (1991)). Typical macro databases store each time series variable as a column (or row)
vector. In the real-time data literature, the remeasurements are recorded as succes-
sive column vectors, and the data for each variable are usually stored as a matrix. The “vintage date” refers to the release date of each vector of time series measure-
ments and the “vintage” denotes the column vector of time series data. Real-time
data comprises many vintages; each successive column vector represents a vintage
containing the data available at that vintage date. The “most recent”, “current”
and “final” labels are used interchangeably to denote the column with the latest
vintage date. These are not the “true” measurements, however, since these will be
revised subsequently too. Some researchers, including Egginton, Pick and Vahey (2002), use successive
vintages (columns) reflecting common practice by applied econometricians in real-
time policy and forecasting analyses. Others, including Howrey (1978) and Koenig
et al (2003), use measurements that have been revised the same number of times
(from the diagonals of the real-time data matrix for a particular variable). Castle and Ellis (2002) provide the most comprehensive UK real-time data set.2
The variables comprise the expenditure components measure of real GDP (known
as GDP(E)) in constant prices: private consumption, investment, government con-
sumption, changes in inventories, exports, imports and GDP(E). The quarterly sea-
sonally adjusted variables were published initially by the Office for National Sta-
tistics (ONS) in Economic Trends and its Annual Supplement. An MS-Excel file
contains separate sheets for each variable. Following the standard conventions in
the literature, the columns reflect the vintages, with time series observations in the
rows. The first vintage refers to 1961Q1 and currently the last refers to 2003Q4.3
Since a typical quarter contains multiple vintages, the frequency of the vintage dates
exceeds the frequency of the time series observations. 2Download from http://www.bankofengland.co.uk/statistics/gdpdatabase.
3Annual updates occur in Spring of each year.
4Download from http://www.econ.cam.ac.uk/dae/keepitreal. Annual updates occur in Spring of each year. 2
Data sources Egginton, Pick and Vahey (2002) provide additional real-time data for: GDP(O)
(output measure of real GDP), private consumption, retail sales, government sur-
plus, unemployment (total claimant count), M0, M3, M4, industrial production and
average earnings.4 The first two quarterly series and the remaining monthly vari-
ables came from the ONS publications Economic Trends and Financial Statistics. Variables are downloadable individually in MS-Excel and ASCII text format. With
the exception of the monetary aggregates, the sequence of vintages starts in January
1980 and ends in June 1999. For the monetary variables, M0, M3 and M4, the first
vintages are June 1981, January 1980 and June 1987 respectively, reflecting avail-
ability in the source publications. All variables are seasonally adjusted except the
budget surplus. Unfortunately, the Egginton-Pick-Vahey data set contains no “deep
history” information. The published versions of the original sources only show a
(moving) window of data at any point in time. Empty cells denote data outside of
that window–generally in excess of two years before the vintage date. One concern for researchers interested in UK monetary issues is the absence One concern for researchers interested in UK monetary issues i 3 of quarterly monetary aggregates.5
To address this omission, we collected real-
time data on quarterly seasonally adjusted M0 and M4 from the ONS’ Economic
Trends for the vintages July 1987 to August 2002. We included (from the same
sources) additional real-time information on nominal GDP(E), GDP price deflator,
M0 velocity and M4 velocity. The interest in money velocity stems from its pivotal
role in the UK’s 1980’s monetary targeting experiments.6 For the last four variables,
the vintages start in November 1981 and end in 2002. Like the Egginton-Pick-Vahey
data set, the absence of deep history results in some empty cells. The Appendix
contains more complete data descriptions.7 The causes of the UK revisions apparent in all three data sets are discussed in
detail by Castle and Ellis (2002) and Mitchell (2004). In brief, revisions occur when
the ONS receive new data, change their methodology or re-base variables. The new
data category sometimes involves the substitution of delayed survey information for
earlier judgement. The changes in methodology, associated with both the major
structural reforms, following the Pickford Report and the Chancellor’s Initiatives
(see Wroe (1993)), and other more minor reforms have unknown implementation
dates. In contrast, the re-basing dates are known, and occur approximately every
five years. 5The monthly seasonally adjusted monetary aggregates contained in Egginton, Pick and Vahey
(2002) were seasonally adjusted on a different basis from the quarterly equivalents for some of the
period.
6 6See for example Jansen (1998). Although money velocities can be constructed from the compo-
nent variables, nominal GDP and the relevant monetary aggregates, we report the official measures
for completeness. 8The “noise” model analysed by Mankiw, Runkle and Shapiro (1984) has the “final” measure-
ments as the explanatory variable.
In this case, the unbiased revisions are orthogonal to final
measurements–the data collection agency remeasures with errors in variables. 2
Data sources Unlike the other variables in our study, the monetary aggregate data were
collected by the Bank of England not the ONS. Topping and Bishop (1989) discuss
the definitions, collection of, breaks in and revisions to UK monetary aggregates. 7The data are available in MS-Excel format on request from a.garratt@bbk.ac.uk.
8 3
Methodology Our basic model for characterising UK remeasurements: Y k
t
= α + βXk
t + k
t ,
t = 1, . . . , T
(1) where Y k
t = XF
t −Xk
t defines the “revisions”, XF
t
denotes the growth rate of the
“final” measurement and Xk
t denotes the kth measurement of the growth rate of the
macro variable, k = 1, . . . , K where K < F. Notice that the preliminary measure-
ment on the right hand side predates the final measurement used to construct the
left hand side variable. The model corresponds to the “news” or “rational forecast”
specification analysed by (among others) Mankiw, Runkle and Shapiro (1984). The
null hypothesis of unbiasedness, α = 0 and β = 0, indicates unpredictable data
revisions. The orthogonality error condition of ordinary least squares ensures that
revision errors are uncorrelated with preliminary measurements.8 Since the index k = 1, . . . , K denotes the successive measurements for each time
series observation, the Xk
t variable is formed from many “vintages”: one data point 5The monthly seasonally adjusted monetary aggregates contained in Egginton, Pick and Vahey
(2002) were seasonally adjusted on a different basis from the quarterly equivalents for some of the
period. 4 is taken from each vintage. In the results that follow, we restrict attention to the
k = 1 case for brevity. Results for the k = 2, . . . , K case can be obtained from the
authors on request.9 For the vector of “final” data, XF
t , we use the vintage available
from the ONS’ Economic Trends, 6 March 2003 (electronic version). A substantial
time interval exits between the respective sample end dates and the final vintage
date to allow revisions to occur.10 Our model could be extended to allow other macro indicators from the same
information set as Xk
t as explanatory variables (see, for example, Swanson and van
Dijk (2004)). Revisions are “efficient” if, and only if, α, β and the coefficients on the
additional explanatory variables are zero. Unfortunately, theory provides no guid-
ance on what other variables might be useful for testing efficiency and unrestricted
searches for predictability undoubtedly result in a degree of data snooping. 11Bai-Perron
Gauss
code
can
be
downloaded
from
http : //econ.bu.edu/perron/code.html.
Swanson and van Dijk (2004) consider structural breaks in US revisions but restrict attention
to just one break. g
(
)
q
y (
y)
10We repeated our analysis reported below treating XK
t
as the final measurements. Although
this limits the number of revisions allowed in each case the results were qualitatively similar. Again,
the tables can be obtained from the authors on request. 9Contact a.garratt@bbk.ac.uk. We set K = 8 (16) for the quarterly (monthly) variables. 3
Methodology In the
absence of a theoretical basis for an examination of the predictability arising from
other variables, we prefer to test for bias–a sufficient (but not necessary) condition
for inefficiency–and test for multiple structural breaks. Given the unknown implementation dates of some wide-ranging reforms to the
UK data reporting processes (see Wroe (1993)), we adopt the methodology proposed
by Bai and Perron (2003a and 2003b) to search for multiple structural breaks of
unknown timing.11 We introduce some additional notations to our basic revisions
equation (1): Y k
t
= αj + βjXk
t + k
t ,
t = Tj−1 + 1, . . . , Tj
(2) for j = 1, ...., m + 1. The linear regression has m breaks (m + 1 regimes) where the
indices (T1, ...., Tm)–the break points–are unknown, with T0 = 0 and Tm+1 = T. So for the one break point case, m = 1 and j = 1, 2, and the pair of estimated
parameters [bα1, bβ1] corresponds to the sample t = 1, ...., T1 and [bα2, bβ2] corresponds
to the sample t = T1 + 1, ...., T. We define a break as a change in at least one of the
parameters α and β. The Bai and Perron (2003a and 2003b) algorithm conducts efficient automated
searches for multiple breaks based on a dynamic programming approach. The re-
searcher chooses a maximum number of candidate breaks, N, and a trimming factor,
τ. Given these inputs the algorithm splits the sample into feasible sub-samples. The
maximised value of the residual sum of squares identifies the candidate breaks for
each number of breaks, n = 1, . . . , N. The researcher tests the null hypothesis of
no structural change against the alternative of many changes by a Sup Wald test. Having identified at least one change, the number of breaks is identified by specify-
ing the null of n = L (1 ≤L < N) changes against L + 1 changes and conducting a
sequence of SupF(L + 1|L) tests. |
The Bai-Perron approach is robust to serial correlation and heteroskedasticity. Minor reforms to statistical reporting procedures could induce the latter and slow 9Contact a.garratt@bbk.ac.uk. We set K = 8 (16) for the quarterly (monthly) variables. K g
(
)
q
y (
y)
10We repeated our analysis reported below treating XK
t
as the final measurements. 4.1
Data For our characterisations of UK data remeasurements we use sixteen variables in
total. The first six variables are from the Castle-Ellis data set and comprise GDP(E),
consumption, investment, government expenditure, exports and imports, all for the
period 1961Q3-1999Q2; the second six are from the Garratt-Vahey data set and
are nominal GDP and the GDP price deflator for 1981Q1-1999Q4, M0 and M4
for 1987Q1-1999Q4, M0 velocity for 1987Q1-1999Q4 and M4 velocity for 1986Q4-
1998Q4; the final four are from the Egginton-Pick-Vahey data set and are average
earnings for 1979M11-1997M1, industrial production for 1979M11-1997M8, claimant
count unemployment for 1979M12-1997M10 and retail sales for 1986M2-1997M12. Since the indicators vary by source and time series frequency, the sample size,
the trimming factor τ (as a proportion of the sample) and the maximum number
of breaks, N, vary.12 We set N = 5 and τ = 0.15 for the 150 plus observations
for both the quarterly GDP(E) components in the Castle-Ellis data set and the
monthly indicators from Egginton-Pick-Vahey.13 For the Garratt-Vahey monetary
aggregates and velocities, where there are 52 or 53 quarterly observations, we set
N = 1 and τ = 0.25. For nominal GDP and the price deflator, there are 76 quarterly
observations and we set N = 2 and τ = 0.25. We use quarterly or monthly growth rates as appropriate throughout the empir-
ical section.14 This approach mitigates the level effects that result from base year
changes (see Patterson and Hervai (1991)). In general, conventional unit root tests
indicate that the variables in equation (2) are stationary, despite the small samples
and the likely presence of structural breaks. Table 1 reports the means and standard deviations of revisions, Y 1
t . In gen-
eral, the mean revisions are positive: preliminary measurements understate final
measurements but there is considerable variation across variables. Approximately
half of the indicators have statistically significant mean revisions at the 5% level
(denoted by ∗in Table 1). Investment has the largest (quarterly) mean revisions:
nearly twice as big as GDP(E).15 The notably small M0 and M4 mean revisions
are insignificantly different from zero at the 5% level. The mean absolute error for
the monetary aggregates is also notably lower than for the other variables. The
preliminary analysis suggest little predictability for monetary aggregate revisions. To illustrate the scale of revisions, Figure 1 plots GDP(E) from 1961Q3 to
1999Q2 for the first and final measurements. 3
Methodology Although
this limits the number of revisions allowed in each case the results were qualitatively similar. Again,
the tables can be obtained from the authors on request. 11Bai-Perron
Gauss
code
can
be
downloaded
from
http : //econ.bu.edu/perron/code.html. Swanson and van Dijk (2004) consider structural breaks in US revisions but restrict attention
to just one break. 5 adjustment by the agency would cause the former (see Barklem (2000)). Our ap-
proach tests the stability of bias allowing for badly-behaved errors. (
)
pp
p
p
p
13The Castle-Ellis data set contains (at times) more than one vintage per quarter. We used the
vintage available at the start of each quarter and treated the Garratt-Vahey variables analogously.
14 14The growth rates for Xt were defined as 100 ∗(loge Xt −loge Xt−1).
15The GDP(E) revisions are comparable in size to those documented by Faust, Rogers and Wright
(2004). 12Bai and Perron (2003b) discuss the appropriate parameter values in small samples.
13The Castle-Ellis data set contains (at times) more than one vintage per quarter. We used the
vintage available at the start of each quarter and treated the Garratt-Vahey variables analogously.
14The growth rates for Xt were defined as 100 ∗(loge Xt −loge Xt−1).
15The GDP(E) revisions are comparable in size to those documented by Faust, Rogers and Wright
(2004). 12Bai and Perron (2003b) discuss the appropriate parameter values in small samples. Bai and Perron (2003b) discuss the appropriate parameter values in small samples.
13The Castle-Ellis data set contains (at times) more than one vintage per quarter. We used the
vintage available at the start of each quarter and treated the Garratt-Vahey variables analogously.
14The growth rates for Xt were defined as 100 ∗(loge Xt −loge Xt−1). The growth rates for Xt were defined as 100
(loge Xt
loge Xt
1).
15The GDP(E) revisions are comparable in size to those documented by Faust, Rogers and Wright
(2004). 16The pre and post-break means were 0.23 (Newey-West coefficient standard error 0.091) and
0.81 (0.204) for exports and 0.02 (0.176) and 0.73 (0.219) for imports.
17 17The sample mid-points defined the break dates for the Garratt-Vahey and Egginton-Pick-Vahey
variables. 19The Newey-West truncation factor was 4; and the serial correlation test was for up to 4th (12th)
order for the quarterly (monthly) data. 18Tables for subsequent measurements (up to two years after the initial measurement) can be
obtained from the authors on request. Except for the monetary aggregates, the data reject the null
hypothesis of unbiasedness for all k at the 1% level. However, the degree but not the direction of
bias varies considerably with k. 4.1
Data The deviation between the two shows 6 the k = 1 revision. At times, these are larger in absolute size than the quarterly
economic growth rate. Figure 1 also shows that the final measurements are much
less volatile post-1989, reflecting the relative stability of the 1990s boom. -6.0
-4.0
-2.0
0.0
2.0
4.0
6.0
8.0
61q3
64q1
66q3
69q1
71q3
74q1
76q3
79q1
81q3
84q1
86q3
89q1
91q3
94q1
96q3
99q1
Quarterly % growth
First Measurement, k=1
Final Measurement
Figure 1: GDP(E) Growth, First and Final Measurements Figure 1: GDP(E) Growth, First and Final Measurements To check for structural change in the mean revision of each variable, we estimated
a restricted version of equation (2) with βj = 0. We used the Bai-Perron method-
ology to identify structural breaks of unknown timing in the intercept. There are
breaks in the means only for exports (1993Q3) and imports (1992Q1).16 (The re-
sults reported in the next section based on unrestricted estimation of equation (2)
suggest that the data reject the βj = 0 restriction and that structural breaks are
much more prevalent.) To investigate time variation in the standard deviations for each GDP(E) compo-
nent, we split the sample into two sub-samples, corresponding approximately to the
1980s and 1990s.17 The results suggest a fairly consistent pattern: lower standard
deviations for the 1990s. For 10 of the 16 variables, the data reject the null hypoth-
esis of equal variances for the two sub-samples at the 5% level using a variance ratio
test (denoted by † in Table 1). We conclude from this preliminary investigation that revisions are often pre-
dictable and typically positive, with considerable variation in size across variables
and lower 1990s’ revision volatility. 7 1961
1970
1980
1990
2000
@
@
R
? Consumption
1967Q2
¡
¡
ª
Government
expenditure
1975Q1
? Imports
1985Q4
? Exports
1971Q4
? Exports
1980Q3
¡
¡
ª
Exports
1993Q3
? M0
1993Q2
-
M0
velocity
1992Q1
6
Average
earnings
1987M11
6
Average
earnings
1992M9
¡
¡
µ
Industrial
production
1986M5
@
@
I
Unemployment
1992M10
Figure 2: Break Points ¡
Government
expenditure
1975Q1 Exports
1980Q3 Imports
1985Q4 onsumption
1967Q2
Exports
1971Q4 ¡
Exports
1993Q3 Exports
1971Q4 ¡
Industrial
production
1986M5 Figure 2: Break Points 4.2
Testing for Bias Tables 2, 3 and 4 summarise the results from our regressions based on equation
(2) using 16 macro indicators for the first measurements (k = 1).18 In each case,
we report the p-value for the Wald test of the null hypothesis for unbiasedness,
α = β = 0, Newey-West heteroskedasticity and serial correlation consistent standard
errors and an LM-test statistic for serial correlation.19 The tables show the bias for
each parameter-stable segment; if there are no structural breaks, we report the
results for the full sample. The break points are also shown on a time line in Figure
2. 20Sargent (1989) considers an agency that filters preliminary measurements of investment based
on a predictable relationship with output. The efficiently transformed data exhibit an apparent
investment accelerator even if the economic relationships do not. 4.2.3
Egginton-Pick-Vahey Variables Table 4 reports the results for the remaining four variables, all taken from the
Eggington-Pick-Vahey data set. Both unemployment and industrial production have
one break (in the early 1990s and mid-1980s, respectively); average earnings has two
breaks (one in the late 1980s, the other in the early 1990s). In contrast, retail sales
exhibits no breaks. In general, the preliminary measurements are downwards biased
predictors of subsequent measurements at their sample means–matching the pat-
tern observed for real-side quarterly indicators. The exceptions are unemployment,
average earnings and industrial production before their respective first breaks. These
sub-samples display unbiasedness at the 15% level and above. All three indicators
exhibit bias at the 1% level for subsequent sub-samples, consistent with statistical
quality degradation. 4.2.1
Castle-Ellis Variables Table 2 reports the results for GDP(E) and its components. Most of these variables
have breaks that pre-date the late 1980s’ and early 1990s’ structural reforms to
ONS practices. The exports break in 1993Q3 coincides with the rebasing of national
accounts. In general, the null hypothesis of α = β = 0 can be rejected at the 1%
level, with variation in the size of the bias across variables. Initial measurements are
unfailingly revised upwards (at the sample means). For example, the estimated α
and β values for GDP(E) (investment) are in the region of 0.4 (0.6) and -0.6 (-0.3)
respectively. This implies preliminary GDP(E) (investment) measurements around
the sample mean (quarterly) output growth of 0.4% (0.5%) would be revised to
nearly 0.6% (0.9%). Nearly all variables subject to structural breaks display bias
before and after the breaks; the absolute values of the coefficients are sometimes
larger post-break. The null hypothesis of unbiased revisions can only be rejected in
one sub-sample: for imports before the mid-1980s’ break. 8 8 4.2.2
Garratt-Vahey Variables Table 3 reports the results for the six Garratt-Vahey nominal variables. With the
exception of the early 1990s’ breaks for M0 and its velocity, these variables show
stability over the period. Although nominal GDP revisions and the GDP price
deflator both exhibit significant bias at the 1% level, the monetary aggregates do not,
with p-values above 10% and smaller coefficients (in absolute value). The narrower
measure, M0, displays bias before the early 1990s’ break. In general, the revisions
to the money velocities are biased at the 1% level–reflecting the predictability of
nominal GDP revisions–with an early 1990s’ break for the narrower measure. 5
Forecasting Case Study Strong revisions predictability gives scope for improving real-time forecast perfor-
mance. To illustrate this, we consider a probability event forecasting exercise. We compute one step ahead out-of-sample forecasts for the evaluation period
1990Q1-1999Q2 using the unrestricted VAR estimated recursively:21 Xm
t = δm +
4
X
i=1
Γm
i Xm
t−i + εm
t
(3) where Xm
t
= (ym
t , pm
t )0, m = 1, F and B. The variables y and p denote quarterly
output growth and inflation (defined using the GDP price deflator). The superscript
m = 1, F and B denotes the set of first, final and bias-adjusted measurements
respectively. We define the bias-adjusted measurements, XB
t , as: where Xm
t
= (ym
t , pm
t )0, m = 1, F and B. The variables y and p denote quarterly
output growth and inflation (defined using the GDP price deflator). The superscript
m = 1, F and B denotes the set of first, final and bias-adjusted measurements
respectively. We define the bias-adjusted measurements, XB
t , as: XB
t = α + (1 + β)X1
t
(4) where X1
t denotes the first measurement. We assume that the forecaster knows the
true values of α and β and that they are equal to the respective sample coeficients
from equation (2).22 where X1
t denotes the first measurement. We assume that the forecaster knows the
true values of α and β and that they are equal to the respective sample coeficients
from equation (2).22 To arrive at our preferred specification for the forecasting VAR, we first tested
for stationarity and then selected the lag order. We could not reject the null of
a unit root in the levels data but could reject the null in first differences at the
5% level using augmented Dickey-Fuller tests (for both first and final measurement
data). We selected the lag order by estimating a sequence of unrestricted VAR(p),
p = 0, 1, 2, ..., 6 models. For the first measurement data, m = 1, the optimal Akaike
Information Criteria selected lag length was zero; but for final data, m = F, the lag
order equalled four. Bearing in mind that unnecessary lags causes inefficiency but
not bias in the OLS estimators, we standardised the lag length at four for first, final
and the bias-adjusted data. The sample start date reflects the availability of real-time GDP price deflator data.
22Real-time (m = 1) GDP growth and GDP price deflator inflation exhibit no breaks (see section
4). The values for bα and bβ for GDP growth are 0.444 and -0.573 and for GDP price deflator inflation
are 0.696 and -0.595 respectively. 4.2.4
Discussion The predictability of revisions indicates the potential for improvements in UK sta-
tistical quality. An agency aiming to minimise revisions could exploit revision pre-
dictability. However, filtering prior to data release can create difficulties for mon-
etary and fiscal control. In the absence of transparency, transformed preliminary
measurements severely complicate inferences about the data generating process (Sar-
gent (1989)).20 A statistical agency may prefer a less direct route to efficient revisions based on
gradual reforms to the quality of surveys and in-house estimates. The UK’s well-
known statistical reforms, associated with the Pickford Report and the subsequent
Chancellor’s Initiatives (see Wroe (1993)), had minor impacts on predictability. As
shown in Figure 2, only five structural breaks occurred in the 1989-1995 period. For unemployment, predictability increased post-break. The monetary aggregates
produced by the Bank of England were unaffected by the reforms to ONS procedures. Both exports and average earnings exhibit statistically significant predictability after
their early 1990s’ breaks. Our preliminary analysis indicated that there was, however, some evidence that
the volatility of revisions fell after the Pickford Report. To check the robustness 9 of this characterisation in the presence of structural breaks, we tested for constant
variances across each break identified by the Bai-Perron approach. Using a variance
ratio test, the null of no difference in the variance can be rejected at the 5%, with re-
visions volatility lower post-break for most cases. The exceptions are unemployment
and average earnings (second break). 21The sample start date reflects the availability of real-time GDP price deflator data. 5
Forecasting Case Study For model evaluation, we consider an economic agent monitoring business cycle
turning points by calculating the probability of above trend output growth. This
is sometimes referred to as “positive momentum” or “above speed limit” growth
in the monetary policy literature (Walsh (2003)). We take the (final data) average
economic growth rate for the evaluation period, 0.52%, as the “trend”, the agent
calculates the probability Pr[ym
t
> 0.52|Ωt−1], for m = 1, F and B where Ωt−1
denotes the information set dated t −1. Confidence intervals are of limited help to 10 our agent because the concern with turning points implies little interest in whether
any particular forecast confidence interval encompass a specific value for output. Garratt et al (2003a) and Clements (2004) discuss in detail the appropriateness of
probability forecasts and their relationships to standard forecast confidence intervals. We compute the probability forecasts by stochastic simulation by the methods
described by Garratt et al (2003b, appendix).23
Figure 3 plots the probabilities
of the event for the three data types. For most of the evaluation period, final
data results in a higher probability of above mean output growth than with first
measurement data. The average difference in probabilities is 11.6 percentage points
(with a standard deviation of 22.4%). Using bias-corrected measurements rather
than first-measurement data reduces considerably the mean (absolute) difference in
forecast probabilities to 4.8 percentage points (with a standard deviation of 21.7%),
although substantial differences remain at times. 0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
90q1
91q1
92q1
93q1
94q1
95q1
96q1
97q1
98q1
99q1
Horizon: 1990q1-1999q2
Probability
First
Final
Bias Adjusted
Figure 3: One Step Ahead Probability of Above-trend Output Growth. Figure 3: One Step Ahead Probability of Above-trend Output Growth. 23To obtain probability forecasts by stochastic simulation we simulate values of Xm(s)
T +1 = bδ
m(s)+
4
X
i=1
bΓm
i Xm(s)
t−i +εm(s)
T +1 Xm(s)
T +1 = bδ
m(s)+
4
X
i=1
bΓm
i Xm(s)
t−i +εm(s)
T +1 where T runs from 1989Q4 to 1999Q1, the parameter estimates vary with each recursion, the
superscript ‘(s)’ refers to the sth replication of the simulation algorithm (s = 1, 2, ....1000) and the
εm(s)
T +1 ’s are drawn using a nonparametric method with replacement. The probability of an event is
then defined as the proportion of the ‘(s)’ simulations the event is forecast to occur. 23To obtain probability forecasts by stochastic simulation we simulate values of 24We also used the “probability integral transform” (PIT) method, due to Rosenblatt (1952) and
discussed in detail by Clements (2004). The two events considered were above-trend output growth
and above-trend inflation, giving 76 probability forecasts and their associated realisations for the 38
quarters from 1990Q1 to 1999Q2. We calculated the probability of observing values no greater than
the actual (final data) values. Under the null hypothesis that the set of density forecasts match the
actual data generating density, the PITs are uniformly distributed U[0,1]. The Kolmogorov-Smirnov
statistics indicate marginal rejection for final data but clear rejection with first measurements at
the 5% significance level. The bias-adjusted measurements indicated marginal rejection at the same
significance level. 5
Forecasting Case Study Garratt et al
(2003b) label this type of uncertainty as the effects of unobserved future shocks. 11 For more formal forecast evaluation, Table 5 reports the proportion of correctly
forecast events, P, the Kuipers score statistic, KS, and the Pesaran-Timmermann
(1992) directional market timing statistic, PT. Table 5:
Evaluation of probability event forecasts Measurements
P
KS
PT
First
55.3%
0.047
0.270
Final
73.7%
0.514
2.865
Bias adjusted
63.2%
0.368
1.428 Measurements
P
KS
PT
First
55.3%
0.047
0.270
Final
73.7%
0.514
2.865
Bias adjusted
63.2%
0.368
1.428 We consider 38 events in total; one event (above trend growth) for each time
period in the 1990Q1 to 1999Q2 evaluation period. We assume that an event can be
correctly forecast if the associated probability forecast exceeds 50 percent. Although
over 70% of events can be correctly forecast using final data, using first measurements
and bias-adjusted measurements reduces the success rate by approximately 19 and
11 percentage points respectively. The Kuipers scores also suggest that bias adjustment improves forecast perfor-
mance. This statistic measures the proportion of above mean growth rates that
were correctly forecast minus the proportion of below mean growth rates that were
incorrectly forecast. The test provides a measure of the accuracy of directional fore-
casts, with high positive numbers indicating high predictive accuracy. Using first
measurements gives a KS of approximately 0.05; bias-adjustment betters this score
by 0.32 – considerably closer to the final data score of 0.51. The PT statistic allows a formal hypothesis of directional forecasting perfor-
mance. As shown in Granger and Pesaran (2000), this hypothesis test uses the
same information as the Kuipers score. Under the null hypothesis that the forecasts
and realisations are independently distributed the PT statistic has a standard nor-
mal distribution. The first measurement data reject the null of no ability to forecast
oberved changes with a probability value of 0.78. Bias-adjustment reduces the prob-
ability value to 0.15 – indicating rejection at the 15% level. Final data give clear
rejection at the 1% level. We conclude that bias adjustment improves probability
forecasting performance for this particular forecasting example.24 12 Although this analysis indicates the scope for exploiting revision predictability,
we emphasise that the variation in predictability across variables and through time
ensures that performance improvement is case specific. Furthermore, the parameters
of equation (2) were assumed to be known by the agent (and identified as the pop-
ulation coefficients). In the presence of structural breaks, parameter learning may
limit the scope for increasing forecast accuracy. Modelling the impacts of bounded
rationality on real-time forecast and policy model performance is an interesting area
for subsequent research. 6
Conclusions By utilising both existing and new sources of real-time data, this paper has charac-
terised the revision processes for 16 UK macro indicators. The main finding–that
the preliminary measurements of UK macro variables are generally biased–confirms
a widely-held suspicion that UK macro measurements are inefficient. Where present,
the bias causes preliminary measurements to understate later measurements (at the
sample means) and structural breaks result in some variation in revisions predictabil-
ity. Monetary aggregates, MO and M4, are typically unbiased (at least, post-break). Using a forecasting probability example, we have demonstrated the potential to im-
prove real-time model performance by utilising bias-adjusted data. 13 13 References [1] Bai, J., and P. Perron (1998) “Estimating and Testing Linear Models with
Multiple Structural Changes”, Econometrica, 66, 47-78. [2] Bai, J., and P. Perron (2003a) “Computation and Analysis of Multiple Struc-
tural Change Models”, Journal of Applied Econometrics, 18, 1-22. [3] Bai, J., and P. Perron (2003b) “Critical Values for Multiple Structural Change
Tests”, Econometrics Journal, 6, 72-78. [4] Barklem, A. (2000) “Revisions Analysis of Initial Estimates of Key Economic
Indicators and GDP Components”, Economic Trends, 556, 31-52. [5] Castle, J. and C. Ellis (2002) “Building a Real-time Database for GDP(E)”,
Bank of England Quarterly Bulletin, February, 42-49. [6] Clements, M.P. (2004) “Evaluating the Bank Of England Density Forecasts of
Inflation”, Economic Journal, 114, 844-866. [7] Croushore, D. and T. Stark (2001) “A Real-time Data Set for Macroeconomists”
Journal of Econometrics, 105, 111-130. [8] Diebold, F. X., and G. D. Rudebusch (1991) “Forecasting Output with the
Composite Leading Index: A Real-Time Analysis”, Journal of the American
Statistical Association, 86, 603-610. [9] Egginton, D.M., A. Pick and S.P. Vahey (2002) “ ‘Keep It Real!’ A Real-time
UK Macro Data Set”, Economics Letters, 77, 15-20. [10] Faust, J., J.H. Rogers and J.H. Wright (2004) “News and Noise in G7 GDP
Announcements”, Journal of Money, Credit and Banking, forthcoming. [11] Garratt,
A.,
K. Lee,
M.H. Pesaran,
and Y. Shin,
Y. (2003a) “Fore-
cast Uncertainties in Macroeconometric Modelling:
An Application to
the UK Economy”, Cambridge University Discussion Paper, available at
http://www.econ.cam.ac.uk/faculty/pesaran. [12] Garratt, A., K. Lee, M.H. Pesaran, and Y. Shin, Y. (2003b) “Forecast Uncer-
tainties in Macroeconometric Modelling: An Application to the UK Economy”,
Journal of American Statistical Association, Applications and Case Studies, 98,
464, 829-838. [13] Granger, C.W.J. and M.H. Pesaran (2000), “Economic and Statistical Measures
of Forecast Accuracy,” Journal of Forecasting, 19, 537-560. [14] Howrey, E.P. (1978) “The Use of Preliminary Data in Econometric Forecast-
ing”, Review of Economics and Statistics, 60, 193-200. [15] Jansen, N. (1998), “The Demand for M0 in the United Kingdom Reconsidered:
Some Specification Issues”, Bank of England Working Paper 83. 14 [16] Koenig, E., S. Dolmas and J. Piger (2003) “The Use and Abuse of ‘Real-time’
Data in Economic Forecasting”, Review of Economics and Statistics, 85, 618-
628. [17] Mankiw, N.G., Runkle, D.E and M.D. Shapiro (1984) “Are Preliminary An-
noucements of the Money Stock Rational Forecasts”, Journal of Monetary Eco-
nomics, 14, 15-27. [18] Mitchell, J. 7
Appendix: Summary of Garratt-Vahey real-time data In this appendix, we describe the real-time data collected specifically for this study
(referred to as the Garratt-Vahey data set in the main text). The data consist
of monthly vintages of nominal macroeconomic variables. Each variable has many
different vintages–reflecting the revisions and updates that occur over time. In
the MS-Excel files, the data are stored as a matrix for each variable. Successive
column vectors of the matrix represent different (more recent) vintages of data; each
contains the most recent measurements available at that vintage date. The data were
collected by examining various issues of Economic Trends, which is published by the
ONS (formally the Central Statistical Office). The figures reported were in the public domain at the end of the month in
question. For each vintage, the observations are identical to those in the relevant
published source. The window length reported by the source publications is affected
by page layout considerations–it varies by variable and by vintage date. Missing
data are recorded as empty cells. The two excel files containing the data described
below, nomY&Pdef.xls and money.xls, are available from the authors on request. In the following section, the definition, source, code, period and relevant notes
are described for each variable. 1. Nominal GDP (Excel file: nomY&Pdef.xls, Spreadsheet: nominal mktp(sa)). Definition:- Gross domestic product at market prices, current price $ Million,
seasonally adjusted. Source:- ONS Economic Trends. Code:- FNAM (from Nov 1981 to Sept 1985), CAOB (from Oct 1985 to Sept
1998) and YBHA (from Oct 1998 onwards). Period:- Monthly vintages from Nov 1981 to August 2002, on quarterly obser-
vations 1976Q1 to 2002Q1. 1. Nominal GDP (Excel file: nomY&Pdef.xls, Spreadsheet: nominal mktp(sa)). 1. Nominal GDP (Excel file: nomY&Pdef.xls, Spreadsheet: nominal mktp(sa)). Definition:- Gross domestic product at market prices, current price $ Million,
seasonally adjusted. 1. Nominal GDP (Excel file: nomY&Pdef.xls, Spreadsheet: nominal mktp(sa)). Definition:- Gross domestic product at market prices, current price $ Million,
seasonally adjusted. Definition:- Gross domestic product at market prices, current price $ Million,
seasonally adjusted. 2. GDP price deflator (Excel file: nomY&Pdef.xls, Spreadsheet: deflator mktp). Definition:- Implied market price deflator (average estimate). 2. GDP price deflator (Excel file: nomY&Pdef.xls, Spreadsheet: deflator mktp). Definition:- Implied market price deflator (average estimate). Source:- ONS Economic Trends. Code:- DJDT (from Nov 1981 to Oct 1998) and YBGB (from Oct 1998 on-
wards). Period:- Monthly vintages from Nov 1981 to August 2002, on quarterly obser-
vations 1976Q1 to 2001Q4. References (2004) Revisions to Economic Statistics, National Institute of Eco-
nomic and Social Research, April. [19] Newey, W.K. and K.D. West (1987) “A Simple Positive Semidefinite, Het-
eroskedasticity and Autocorrelation Consistent Covariance Matrix”, Economet-
rica, 55, 703-708. [20] Patterson, K. D., and S. M. Heravi (1991) “Data Revisions and the Expenditure
Components of GDP” Economic Journal, 101, 887-901. [21] Pesaran, M.H. and A. Timmermann (1992), “A Simple Nonparametric Test
of Predictive Performance,” Journal of Business and Economic Statistics, 10,
461-465. [22] Pickford S. (1989) Government Economic Statistics: A Scrutiny Report, Cabi-
net Office, HMSO. [23] Rosenblatt, M. (1952) “Remarks on a Multivariate Transformation”, Annals of
Mathematical Statistics, 23, 470-472. [24] Sargent, T., (1989) “Two Models of Measurements and the Investment Accel-
erator”, Journal of Political Economy, 97, 251-287. [25] Swanson, N.R. and D. Van Dijk (2004) “Are Statistical Reporting Agencies Get-
ting It Right? Data Rationality and Business Cycle Asymmetry”, Econometric
Institute Report EI 2001-28, revised 2004. [26] Symons, P. (2001) “Revisions Analysis of Initial Estimates of Annual Constant
Price GDP and Its Components”, Economic Trends, 568, 48-65. [27] Topping, S.L. and S. L. Bishop (1989), “Breaks in Monetary Series” Bank of
England Discussion Paper, Technical Series, No. 23. [28] Walsh, (2003) “Speed Limit Policies: the Output Gap and Optimal Monetary
Policy”, Amercian Economic Review, 93, 265-278. [29] Wroe, D. (1993) “Improving Macro-economic Statistics”, Economic Trends,
471, 191-199. 15 15 7
Appendix: Summary of Garratt-Vahey real-time data 3. M0 money (Excel file: money.xls, spreadsheet: M0 sa). Definition:- M0, $ Million, Amount outstanding, seasonally adjusted. Source:- ONS Economic Trends. Code:- AVAE. Period:- Monthly vintages from July 1987 to August 2002, on quarterly obser-
vations 1983Q1 to 2002Q1. 16 4. M4 money (Excel file: money.xls, spreadsheet: M4 sa). Definition:- M4, $ Million, Amount outstanding, seasonally adjusted. Source:- ONS Economic Trends. Code:- AUYN. Period:- Monthly vintages from July 1987 to August 2002, on quarterly obser-
vations 1983Q1 to 2002Q1. 5. VM0 money (Excel file: money.xls, spreadsheet: V(M0)). Definition:- Velocity of circulation. Source:- ONS Economic Trends. Code:- AVAM. Period:- Monthly vintages from July 1987 to August 2002, on quarterly obser-
vations 1983Q1 to 2002Q1. Source:- ONS Economic Trends. 6. VM4 money (Excel file: money.xls, spreadsheet: V(M4)). Definition:- Velocity of circulation. Source:- ONS Economic Trends. Code:- AUYU. Period:- Monthly vintages from July 1987 to August 2002, on quarterly obser-
vations 1983Q1 to 2002Q1. 17 17 Table 1: Summary Statistics for Revisions, Y 1
t Sample
Mean
MAE
SD
SD 1990s
GDP(E):
1961Q3 1999Q2
0.24∗
0.88
1.20†
0.31
Consumption:
1961Q3 1999Q2
0.10∗
0.72
0.95†
0.48
Investment:
1961Q3 1999Q2
0.49∗
1.87
2.40†
1.71
Government expenditure:
1961Q3 1999Q2
−0.07
0.96
1.32
1.07
Exports:
1961Q3 1999Q2
0.32∗
1.45
1.80†
1.52
Imports:
1961Q3 1999Q2
0.16
1.44
1.84†
1.33
Nominal GDP:
1981Q1 1999Q4
0.29∗
0.56
0.70
0.64
GDP deflator:
1981Q1 1999Q4
0.07
0.62
0.79†
0.64
M0:
1987Q1 1999Q4
0.05
0.36
0.52†
0.29
M4:
1987Q1 1999Q4
−0.01
0.26
0.35
0.31
M0 velocity:
1987Q1 1999Q4
0.25∗
0.61
0.71†
0.40
M4 velocity:
1986Q4 1999Q4
0.39∗
0.54
0.68
0.67
Average earnings:
1979M11 1997M1
0.03
0.49
0.68
0.77
Industrial production:
1979M11 1997M8
0.05
0.68
0.93†
0.72
Unemployment:
1979M12 1997M10
0.02
0.41
0.61
0.72
Retail sales:
1986M2 1997M12
0.04
0.56
0.73†
0.60
Notes: The revisions, Y 1
t , are defined as the final measurement, XF
t , minus the first
measurement, X1
t . Each measurement, Xt, refers to the quarter-on-quarter (first
12 variables) or month-on-month (last 4 variables) growth rate in percent. MAE is
the mean absolute error; SD refers to standard deviation and SD 1990s refers to the
standard deviation for the 1990s. The symbol ∗denotes statistical significance at
the 5% level using a Newey-West corrected t-statistic based on a regression of the
revision on a constant. Notes: Revisions regression, Equation (2), Y 1
t
= αj + βjX1
t + 1
t .
Newey-West
standard errors (truncation factor equals 4) are in parentheses. We report p-values
of the Wald-test for α = β = 0 and the LM-test statistic for up to 4th-order serial
correlation. 7
Appendix: Summary of Garratt-Vahey real-time data Significantly lower variance for the 1990s at the 5% level
using a variance ratio test is denoted by † (for exact break dates see main text). Notes: The revisions, Y 1
t , are defined as the final measurement, XF
t , minus the first
measurement, X1
t . Each measurement, Xt, refers to the quarter-on-quarter (first
12 variables) or month-on-month (last 4 variables) growth rate in percent. MAE is
the mean absolute error; SD refers to standard deviation and SD 1990s refers to the
standard deviation for the 1990s. The symbol ∗denotes statistical significance at
the 5% level using a Newey-West corrected t-statistic based on a regression of the
revision on a constant. Significantly lower variance for the 1990s at the 5% level
using a variance ratio test is denoted by † (for exact break dates see main text). 18 Table 2:
Revisions regressions, Castle-Ellis Table 2:
Revisions regressions, Castle-Ellis Sample
α
β
R2
Wald-test
LM-test
GDP(E):
1961Q3 1999Q2
0.444
-0.573
0.58
0.00
0.30
(0.065)
(0.050)
Consumption:
1961Q3 1967Q2
0.478
-0.682
0.64
0.00
0.86
(0.120)
(0.087)
1967Q3 1999Q2
0.252
-0.288
0.20
0.00
0.00
(0.072)
(0.051)
Investment:
1961Q3 1999Q2
0.563
-0.320
0.17
0.00
0.01
(0.147)
(0.074)
Government
1961Q3 1975Q1
0.421
-0.591
0.24
0.01
0.17
expenditure:
(0.185)
(0.182)
1975Q2 1999Q2
0.181
-0.816
0.33
0.00
0.01
(0.083)
(0.128)
Exports:
1961Q3 1971Q4
0.630
-0.237
0.43
0.00
0.13
(0.117)
(0.032)
1972Q1 1980Q3
0.219
-0.068
0.01
0.34
0.03
(0.156)
(0.077)
1980Q4 1993Q3
0.495
-0.525
0.45
0.00
0.00
(0.139)
(0.063)
1993Q4 1999Q2
1.274
-0.458
0.25
0.00
0.43
(0.290)
(0.159)
Imports:
1961Q3 1985Q4
0.156
-0.120
0.05
0.03
0.00
(0.130)
(0.043)
1986Q1 1999Q2
0.920
-0.473
0.42
0.00
0.67
(0.290)
(0.082)
1
1
1 19 Notes: Revisions regression, Equation (2), Y 1
t
= αj + βjX1
t + 1
t .
Newey-West
standard errors (truncation factor equals 4) are in parentheses. We report p-values
of the Wald-test for α = β = 0 and the LM-test statistic for up to 4th-order serial
correlation. Table 3:
Revisions regressions, Garratt-Vahey Sample
α
β
R2
Wald-test
LM-test
Nominal GDP:
1981Q1 1999Q4
0.929
-0.431
0.27
0.00
0.22
(0.141)
(0.097)
GDP deflator:
1981Q1 1999Q4
0.696
-0.595
0.30
0.00
0.16
(0.135)
(0.078)
M0:
1987Q1 1993Q2
0.555
-0.494
0.50
0.00
0.09
(0.131)
(0.078)
1993Q3 1999Q4
0.114
-0.035
-0.03
0.46
0.08
(0.077)
(0.041)
M4:
1987Q1 1999Q4
-0.020
0.006
-0.02
0.97
0.16
(0.081)
(0.030)
M0 velocity:
1987Q1 1992Q1
0.937
-1.040
0.72
0.00
0.97
(0.049)
(0.190)
1992Q2 1999Q4
0.025
-0.538
0.48
0.00
0.26
(0.074)
(0.114)
M4 velocity:
1986Q4 1999Q4
0.254
-0.141
0.04
0.00
0.91
(0.111)
(0.80)
Notes: Revisions regression, Equation (2), Y 1
t
= αj + βjX1
t + 1
t . Newey-West
standard errors (truncation factor equals 4) are in parentheses. We report p-values
of the Wald-test for α = β = 0 and the LM-test statistic for up to 4th-order serial
correlation. 20 Table 4:
Revisions regressions, Egginton-Pick-Vahey Table 4:
Revisions regressions, Egginton-Pick-Vahey g
,
gg
y
Sample
α
β
R2
Wald-test
LM-test
Average earnings:
1979M11 1987M11
0.108
-0.138
0.05
0.23
0.00
(0.072)
(0.079)
1987M12 1992M9
0.464
-0.688
0.45
0.00
0.01
(0.091)
(0.129)
1992M10 1997M1
0.257
-0.907
0.87
0.00
0.19
(0.028)
(0.045)
Industrial
1979M11 1986M5
0.050
-0.033
-0.01
0.80
0.06
production:
(0.081)
(0.076)
1986M6 1997M8
0.103
-0.508
0.29
0.00
0.00
(0.047)
(0.092)
Unemployment:
1979M12 1992M10
0.063
-0.026
0.00
0.42
0.00
(0.056)
(0.30)
1992M11 1997M10
-0.428
-0.328
0.25
0.01
0.13
(0.154)
(0.101)
Retail sales:
1986M2 1997M12
0.120
-0.390
0.41
0.00
0.00
(0.029)
(0.042)
Notes: Revisions regression, Equation (2), Y 1
t
= αj + βjX1
t + 1
t . Newey-West
standard errors (truncation factor equals 4) are in parentheses. We report p-values
of the Wald-test for α = β = 0 and the LM-test statistic for up to 12th-order serial
correlation. Sample
α
β
R2
Wald-test
LM-test
Average earnings:
1979M11 1987M11
0.108
-0.138
0.05
0.23
0.00
(0.072)
(0.079)
1987M12 1992M9
0.464
-0.688
0.45
0.00
0.01
(0.091)
(0.129)
1992M10 1997M1
0.257
-0.907
0.87
0.00
0.19
(0.028)
(0.045)
Industrial
1979M11 1986M5
0.050
-0.033
-0.01
0.80
0.06
production:
(0.081)
(0.076)
1986M6 1997M8
0.103
-0.508
0.29
0.00
0.00
(0.047)
(0.092)
Unemployment:
1979M12 1992M10
0.063
-0.026
0.00
0.42
0.00
(0.056)
(0.30)
1992M11 1997M10
-0.428
-0.328
0.25
0.01
0.13
(0.154)
(0.101)
Retail sales:
1986M2 1997M12
0.120
-0.390
0.41
0.00
0.00
(0.029)
(0.042)
Notes: Revisions regression, Equation (2), Y 1
t
= αj + βjX1
t + 1
t . Newey-West
standard errors (truncation factor equals 4) are in parentheses. We report p-values
of the Wald-test for α = β = 0 and the LM-test statistic for up to 12th-order serial
correlation. Notes: Revisions regression, Equation (2), Y 1
t
= αj + βjX1
t + 1
t . Newey-West
standard errors (truncation factor equals 4) are in parentheses. We report p-values
of the Wald-test for α = β = 0 and the LM-test statistic for up to 12th-order serial
correlation. Notes: Revisions regression, Equation (2), Y 1
t
= αj + βjX1
t + 1
t . Newey-West
standard errors (truncation factor equals 4) are in parentheses. We report p-values
of the Wald-test for α = β = 0 and the LM-test statistic for up to 12th-order serial
correlation. Notes: Revisions regression, Equation (2), Y 1
t
= αj + βjX1
t + 1
t . Table 4:
Revisions regressions, Egginton-Pick-Vahey Newey-West
standard errors (truncation factor equals 4) are in parentheses. We report p-values
of the Wald-test for α = β = 0 and the LM-test statistic for up to 12th-order serial
correlation. 21
|
W4392400906.txt
|
https://journal.unindra.ac.id/index.php/hortatori/article/download/2022/1522
|
id
|
Bentuk Kesalahan Berbahasa pada Pidato Mahasiswa yang Berperan sebagai Kepala Desa Terpilih
|
Hortatori
| 2,023
|
cc-by-sa
| 4,207
|
Hortatori
Volume 7 Nomor 2 (2023), 133-140
ISSN 2579-7832 (Cetak) | ISSN 2579-7840 (Elektronik)
Jurnal Pendidikan Bahasa dan Sastra Indonesia
Artikel Penelitian
Bentuk Kesalahan Berbahasa pada Pidato Mahasiswa yang Berperan sebagai
Kepala Desa Terpilih
Dewi Sartika1*)
Atiqah Sabardila2
Universitas Muhammadiyah Surakarta1, 2
*) Penulis Korespondensi: Jl. A. Yani Tromol Pos 1 Pabelan Kartasura, Surakarta, Jawa Tengah, 0271, Indonesia.
Posel: s200220010@student.ums.ac.id, Atiqa.Sabardila@ums.ac.id.
Abstrak: Tujuan dari penelitian ini adalah untuk mengarakterisasi kesalahan linguistik yang dilakukan
oleh mahasiswa MPBI-UMS saat memerankan kepala desa terpilih. Penelitian ini menggunakan strategi
deskriptif kualitatif. Kata, frase, klausa, dan kalimat dari pidato penerimaan yang diberikan oleh
mahasiswa MPBI-UMS setelah terpilih menjadi kepala desa dimasukkan dalam kumpulan data untuk
dianalisis. Wacana lisan siswa berfungsi sebagai sumber data, sedangkan metodologi mendengarkan
dan mencatat berfungsi sebagai teknik pengumpulan data. Analisis padanan referensial, analisis
padanan fonetik artikulatoris, analisis perluasan dalam metode agih, dan analisis pembacaan tanda
digunakan untuk menganalisis data. Berikut adalah temuan dari investigasi ini yaitu terdapat 20 data
bentuk kesalahan berbahasa yang meliputi: (1) Kesalahan pada bidang fonologi: Ditemukan 5 data
bentuk kesalahan berbahasa, termasuk kesalahan dalam penulisan, penghilangan fonem vokal,
kesalahan dalam penulisan huruf kapital, dan kesalahan dalam penulisan cetak miring. (2) Kesalahan
pada bidang morfologi: Terdapat 1 data bentuk kesalahan berbahasa, yaitu pengulangan kata atau
reduplikasi. (3) Kesalahan pada bidang sintaksis: Terdapat 12 data bentuk kesalahan berbahasa, yang
meliputi kesalahan dalam penulisan kalimat atau kata mubadzir, penggunaan kata tidak baku, dan
kalimat tidak jelas. (4) Kesalahan ejaan: Terdapat 2 data bentuk kesalahan ejaan, yakni kesalahan dalam
penggunaan tanda baca. Kesalahan berbahasa bisa terjadi dikarenakan kurangnya pemahaman penutur
terhadap kaidah bahasa Indonesia yang baik dan benar. Melalui pembelajaran bahasa yang rutin,
kompetensi berbahasa seseorang dapat diperbaiki menjadi lebih baik.
Kata kunci: Kesalahan Berbahasa; Mahasiswa; Berpidato.
Forms of Language Errors in the Speeches of Students Who Play the Role of
Elected Village Heads
Abstract: The purpose of this research is to characterize linguistic mistakes made by MPBI-UMS
students when playing the role of the elected village head. This study uses a qualitative descriptive
strategy. Words, phrases, clauses, and sentences from the acceptance speeches given by MPBI-UMS
students after being elected as village heads were included in the data set for analysis. The students'
oral discourse serves as a data source, while the listening and note-taking methodology functions as a
data collection technique. Referential equivalent analysis, articulatory phonetic equivalent analysis,
expansion analysis in agile method, and sign reading analysis were used to analyze the data. The
following are the findings of this investigation, namely that there are 20 data on forms of language
errors which include: (1) Errors in the field of phonology: Found 5 data on forms of language errors,
including errors in writing, omission of vowel phonemes, errors in writing capital letters, and errors in
writing italic. (2) Errors in morphology: There is 1 data in the form of language errors, namely word
repetition or reduplication. (3) Errors in syntax: There are 12 data in the form of language errors,
which include errors in writing redundant sentences or words, use of non-standard words, and unclear
sentences. (4) Spelling errors: There are 2 data in the form of spelling errors, namely errors in the use
of punctuation marks. Language errors can occur due to a lack of understanding of speakers of good
and correct Indonesian language rules. Through routine language learning, one's language competence
can be improved for the better.
Keywords: Language Errors; Student; Speech.
133
134
Bentuk Kesalahan Berbahasa pada Pidato Mahasiswa yang Berperan sebagai Kepala Desa Terpilih
Proses artikel: Dikirim: 19-07-2023; Direvisi: 29-11-2023; Diterima: 29-11-2023; Diterbitkan: 31-12-2023
Gaya sitasi (MLA edisi ke-7): Sartika, Dewi, and Atiqah Sabardila. “Bentuk Kesalahan Berbahasa pada Pidato
Mahasiswa yang Berperan sebagai Kepala Desa Terpilih.” Hortatori: Jurnal Pendidikan Bahasa dan Sastra Indonesia 7.2
(2023): 133–140. Print/Online. Pemegang Hak Cipta: Dewi Sartika, Atiqah Sabardila. Publikasi Utama: Hortatori:
Jurnal Pendidikan Bahasa dan Sastra Indonesia (2023).
Proses ini berada di bawah lisensi Creative Commons Attribution-ShareAlike 4.0 International License.
Pendahuluan
Pidato adalah salah satu bentuk komunikasi yang penting, terutama bagi pemimpin seperti kepala desa.
Pemahaman yang baik tentang bahasa dan kemampuan berbicara yang baik adalah kualitas penting bagi
seorang pemimpin dalam berkomunikasi dengan masyarakat, mengekspresikan visi, dan memimpin dengan
efektif. Seorang kepala desa adalah figur publik yang memiliki peran penting dalam masyarakat. Kesalahan
berbahasa dalam pidato bisa berdampak negatif terhadap reputasi dan kredibilitas seorang kepala desa.
Penelitian ini dapat membantu mendeteksi dan mengurangi kesalahan berbahasa yang dapat merusak citra
pemimpin dan mempengaruhi persepsi masyarakat. Pidato seorang kepala desa dapat mengandung
informasi tentang kebijakan, program, dan keputusan yang akan diambil. Jika pidato tersebut tidak
disampaikan dengan baik atau mengandung kesalahan berbahasa, informasi penting bisa salah dipahami
atau merugikan masyarakat. Penelitian ini dapat membantu memastikan bahwa pesan yang disampaikan
dalam pidato benar-benar diterima dengan baik oleh masyarakat. Mahasiswa yang memerankan peran
kepala desa terpilih dalam penelitian ini dapat memanfaatkan kesempatan ini sebagai latihan untuk
mengembangkan kemampuan berbicara, pemahaman tentang bahasa yang baik, serta kemampuan
berkomunikasi yang efektif. Pengalaman ini dapat membantu mahasiswa mempersiapkan diri untuk peran
kepemimpinan di masa depan.
Pada hakikatnya, tuturan merupakan suatu pengucapan yang menghasilkan bunyi berupa kalimat
yang berisi ide pikiran seseorang yang ditujukan kepada orang lain (Nur Qoyyimah & Sabardila, 2021).
Mahasiswa memiliki kemampuan berbahasa yang termasuk dalam aktivitas berpidato impromptu, di mana
mereka berperan sebagai kepala desa terpilih dalam situasi yang tidak direncanakan sebelumnya. Pidato
merupakan keterampilan berbahasa yang dapat mengukur dan menggambarkan kemampuan seseorang
dalam menguasai bahasa tertentu yang masih kurang (Yana et al., 2021). Selanjutnya, dalam berbahasa
terdapat kekeliruan atau penyimpangan bentuk bahasa yang diucapkan seseorang dalam berdialog atau
berpidato. Fenomena kesalahan berbahasa dapat terjadi dalam situasi atau bidang-bidang tertentu, terutama
dalam penggunaan bahasa yang tidak hanya mengutamakan faktor komunikatif sebagai hasil akhir dalam
kegiatan berbahasa, tetapi juga memperhatikan kaidah berbahasa (Johan et al., 2018).
Kesalahan berbahasa dalam berpidato merupakan bentuk kesalahan bahasa dalam bidang
linguistik. Bentuk-bentuk kesalahan berbahasa dalam kategori ilmu bidang linguistik meliputi bidang
fonologi, bidang morfologi, sintaksis, dan kesalahan penulisan ejaan. Dalam penelitian yang dilakukan oleh
Sikana et al. (2021) yang mengkaji "Kesalahan Berbahasa Tataran Fonologi pada Pidato Juru Bicara
Penanganan Virus Covid-19 Achmad Yurianto", Dalam penelitian tersebut, ditemukan tiga tipe kesalahan
bahasa pada tingkat fonologi. Ketiga kesalahan tersebut termasuk penghilangan bunyi, baik vokal,
konsonan, atau keduanya, penambahan bunyi, baik vokal atau konsonan, serta perubahan bunyi yang
melibatkan perubahan bunyi vokal dan konsonan.
Beberapa penelitian terdahulu yang relevan yang telah dilakukan yaitu oleh Hasanudin (2017),
(Reistanti, 2017), (Fajarwati, 2017), (Prihantoro, 2019), (Saputri, 2019), (Agustina & Oktaviana, 2019),
(Gani & Arsyad, 2019), (Busyro, 2020), (Syafi’i et al. 2021), dan (Giawa, 2022). Berdasarkan beberapa
studi penelitian sebelumnya yang relevan, temuan penelitian dominan ditemukan khususnya kesalahan
fonetik, morfologi, sintaksis, dan ortografi. Penelitian ini menemukan kesalahan morfologi meliputi
awalan, akhiran, dan duplikasi/pembentukan ulang. Kesalahan sintaksis termasuk kalimat yang tidak jelas,
salah struktur kalimat, dan tidak efisien penggunaan kalimat. Kesalahan fonologis kapitalisasi dan
kombinasi kata. Menurut pola kesalahan dalam tuturan siswa yang bertindak sebagai Bupati terpilih di
kabupaten Blora, ditemukan kesalahan bahasa di berbagai bidang morfologi, sintaksis, ortografi, dan
Hortatori | Jurnal Pendidikan Bahasa dan Sastra Indonesia | Vol. 7 No. 2 (2023), 133-140
ISSN 2579-7832 (Cetak) | ISSN 2579-7840 (Elektronik) | Url: https://journal.unindra.ac.id/index.php/hortatori/index
Dewi Sartika, Atiqah Sabardila
135
fonologi. Adapun letak perbedaan antara penelitian ini dengan penelitian terdahulu ialah pada objek
penelitian yang digunakan.
Berdasarkan hasil penelitian sebelumnya, penulis menyimpulkan bahwa kesalahan dalam
menggunakan bahasa adalah aspek yang menarik dan layak untuk diteliti lebih lanjut. Oleh karena itu,
penelitian ini diberi judul "Analisis Bentuk Kekeliruan Bahasa pada Pidato Mahasiswa yang Berperan
Sebagai Kepala Desa Terpilih." Tujuan utama dari penelitian ini adalah untuk melakukan deskripsi
menyeluruh mengenai berbagai jenis kekeliruan bahasa yang terdapat dalam pidato, terutama pada aspek
linguistik yaitu fonologi, morfologi, sintaksis, ejaan.
Metode
Penelitian ini merupakan jenis penelitian kualitatif dengan pendekatan deskriptif. Dalam penelitian
deskriptif kualitatif, peneliti bertujuan untuk menguraikan dan menjelaskan data hasil penelitian. Fokus
utama dari penelitian ini adalah untuk menggambarkan atau menjelaskan temuan analisis data yang
diperoleh dalam penelitian, dengan pendekatan yang alami atau sesuai dengan kondisi alam (Sugiyono,
2012). Kalimat dari transkrip ucapan digunakan untuk menyusun kumpulan data ini. Satu orang mahasiswa
Magister Pendidikan Bahasa Indonesia Universitas Muhammadiyah Surakarta yang terpilih sebagai kepala
desa di Desa Manang memberikan data penelitian melalui sambutan dadakan. Tujuan utama dari penelitian
ini adalah untuk memberikan analisis komprehensif tentang berbagai kategori kesalahan linguistik yang
ada dalam tuturan, termasuk namun tidak terbatas pada pengucapan, morfologi, sintaksis, dan ejaan.
Mendengarkan dan mencatat digunakan sebagai sarana untuk mengumpulkan informasi.
Dalam penelitian ini, digunakan metode analisis data untuk mengidentifikasi permasalahan yang
akan diteliti. Metode analisis ini mencakup teknik dalam metode padan, seperti padan fonetis.artikulatoris
dan.padan.referensial. Metode padan referensial.digunakan untuk menemukan kesalahan bahasa dalam
elemen pengisi kalimat. Teknik dasar yang diterapkan dalam metode padan adalah teknik Pilah Unsur
Penentu (PUP), yang bertumpu pada padan referensial. Selain itu, metode agih juga digunakan dengan
menerapkan teknik perluasan. Teknik perluasan ini berfungsi untuk memperluas pemahaman terhadap data
yang ada. Secara ringkas, penelitian.ini.menggunakan beragam..metode analisis..data, termasuk
metode.padan.dan metode..agih, serta teknik Pilah Unsur Penentu (PUP) dan teknik perluasan. Hal ini
bertujuan untuk menggali lebih dalam dan mengungkapkan informasi mengenai permasalahan yang sedang
diteliti dengan cara kreatif dan inovatif dalam menganalisis satuan bahasa. Pendekatan pembacaan tanda
juga digunakan untuk analisis. Metode pembacaan tanda ini dilakukan dengan memeriksa data penanda
secara mendalam. Di sini dilakukan analisis sintaksis dan morfologis (Sudaryanto, 2013). Gunakan teori
triangulasi untuk memverifikasi keandalan data yang dikumpulkan untuk penyelidikan ini.
Hasil dan Diskusi
Jenis kesalahan bahasa dalam penelitian ini meliputi berbagai bidang kajian, khususnya bidang fonologi,
morfologi, sintaksis, dan ejaan. Di sini, setiap domain kesalahan bahasa dijelaskan sebagai berikut:
Tabel..1 Tabulasi data kesalahan berbahasa pada pidato impromptu mahasiswa MPBI-UMS.
No
Data Kesalahan Berbahasa pada Transkrip Tuturan
Pidato Mahasiswa MPBI-UMS
Jenis Kesalahan
Berbahasa
Jumlah
Kesalahan
1
2
Bidang Fonologi
Bidang Morfologi
5
1
3
4
Bidang Sintaksis
Bidang Ejaan
12
2
Kesalahan Fonologi
Kesalahan bahasa yang berkaitan dengan kajian fonologi dalam penelitian ini meliputi kesalahan perubahan
lambang bunyi, konstruksi fonem, pengucapan, dan kapitalisasi (Setyawati, 2013:24; Markhamah &
Sabardila, 2014:87; Kim, 2015:172; Thoyib dan.Hamidah, 2018:69). Sementara itu, data tentang kesalahan
fonologis dalam tuturan adalah sebagai berikut:
Hortatori | Jurnal Pendidikan Bahasa dan Sastra Indonesia | Vol. 7 No. 2 (2023), 133-140
ISSN 2579-7832 (Cetak) | ISSN 2579-7840 (Elektronik) | Universitas Indraprasta PGRI
136
Bentuk Kesalahan Berbahasa pada Pidato Mahasiswa yang Berperan sebagai Kepala Desa Terpilih
(1) Yang saya hormati kepala urusan, kepela seksi, serta staf pemerintahan Desa Manang,
Kecamatan Grogol, Kabupaten Sukoharjo.
(2) Organisasi kepemudaan di Desa Manang beserta hadirin sekalian yang berbahagi.
(3) Selanjutnya, saya selaku kepala desa tdi Desa Manang ini segenap jajaran yang akan
membersamai dalam memimpin pemerintah Desa Manang ini.
(4) Puji syukur marilah kita panjatkan kehadirat Allah Subhanahu Wa Ta’ala Tuhan yang maha
kuasa sehingga kita bisa berkumpul pada siang hari ini dalam keadaan sehat.
(5) Dengan pengalihan dari TV analog menuju ke TV digital ada juga di desa kita beberapa warga
mendapatkan bantuan set top box tersebut.
Jenis kesalahan bahasa pada data (1) sampai data (3) adalah kesalahan bahasa menghilangkan
fonem. Ejaan kata kepela pada data (1), seharusnya dieja atau dibaca kepala, ada vokal /a/ yang hilang.
Kata kepala pada kalimat data (1) merupakan sebagai tanda penghormatan seseorang yang memiliki jabatan
yang lebih terpandang dari pada orang lain. Penulisan kata berbahagi pada data (2) seharusnya ditulis
berbahagia, terjadi penghilangan fonem vokal /a/. Kata berbahagia pada kalimat data (2) sebagai tanda hari
yang penuh suka cita yang ditujukan kepada seluruh warga Desa. Penulisan kata tdi pada data (3)
seharusnya ditulis tadi, terjadi penghilangan fonem vokal /a/. Kata tadi pada kalimat data (3) menerangkan
waktu telah berlalu.
Selanjutnya kesalahan penggunaan huruf kapital juga merupakan kesalahan berbahasa pada bidang
fonologi. Pada penulisan maha kuasa pada kalimat data (4), seharusnya Maha Kuasa ditulis dengan di awal
kata dengan huruf kapital /M/ dan /K/. Kata Maha Kuasa pada kalimat data (4) memiliki arti bahwa kata
Maha Kuasa merupakan sebagai tanda mengagungkan Tuhan yang memiliki derajat yang maha tinggi.
Kesalahan penggunaan cetak miring pada bahasa asing juga terdapat pada kalimat data (5) yaitu pada kata
TV anallog dan sett top box, seharusnya TV analog dan set top box ditulis dengan di cetak miring karena
sebagai tanda bahasa asing. Temuan penelitian ini dapat diterapkan pada penelitian sebelumnya yang
dilakukan oleh (Ariesta & Sabardila, 2021) yang terkait erat dengan penghilangan fonem, penyertaan atau
kelebihan fonem, kesalahan penggunaan huruf besar, kesalahan penggunaan huruf miring. Sedangkan pada
penelitian terkait dengan kesalahan penggunaan huruf kapital dan kesalahan penggunaan penulisan cetak
miring.
Kesalahan Bidang Morfologi
Morfologi merupakan ilmu bahasa yang mengkaji morfem atau kata, yaitu mengidentifikasi satuan-satuan
yang membentuk kata menjadi satuan gramatikal. Proses morfologi yang disebut juga afiksasi, perulangan
reduplikasi, dan pemajemukan pada hakikatnya ialah proses pembentukan kata dari struktur dasar dengan
penambahan afiks. (Diari, 2018). Adapun, data pada kalimat yang terdapat dalam pidato yaitu:
(6) Yang terhormat Camat Kecamatan Grogol, Kabupaten Sukoharjo.
Kesalahan berbahasa pidato pada data (6) tersebut merupakan reduplikasi bentuk ulang kalimat.
Kalimat tersebut berbunyi : “Yang terhormat Camat Kecamatan Grogol, Kabupaten Sukoharjo. Kalimat
pada kutipan tersebut merupakan kesalahan berbahasa dalam bidang morfologi. Seharusnya cukup
menggunakan kaliamat: yang terhormat camat Grogol. Dengan demikian, dalam kalimat "Yang terhormat
Camat Kecamatan Grogol, Kabupaten Sukoharjo," reduplikasi kata "Camat" dan "Kecamatan" memiliki
peran penting dalam memberikan penekanan, nuansa formalitas, dan informasi yang jelas terkait jabatan
dan wilayah administratif yang dihormati. Penelitian yang setopik dengan kesalahan bentuk ulang
dilakukan oleh Sintia et. al (2019) terdapat kesalahan bentuk ulang sayur sayur. Bentuk benar dalam
penggunaan bentuk ulang tersebut yaitu sayur mayur.
Kesalahan Bidang Sintaksis
Dalam penelitian tentang kesalahan bahasa dalam ranah sintaksis, penelitiannya meliputi beberapa hal
penting, seperti urutan kata, kesesuaian kata, penyusunan frasa, kesesuaian kalimat, dan logika kalimat
(Markhamah dan Sabardila, 2014: 137). Temuan penelitian menunjukkan bahwa terdapat kesalahan bahasa
yang ditemukan dalam pidato, terutama dalam aspek sintaksis sebagai berikut.
(7) Yang terhormat Camat Kecamatan Grogol, Kabupaten Sukoharjo.
Hortatori | Jurnal Pendidikan Bahasa dan Sastra Indonesia | Vol. 7 No. 2 (2023), 133-140
ISSN 2579-7832 (Cetak) | ISSN 2579-7840 (Elektronik) | Url: https://journal.unindra.ac.id/index.php/hortatori/index
Dewi Sartika, Atiqah Sabardila
137
(8) Pada kesempatan ini saya selaku kepala desa terpilih di Desa Manang Kecamatan Grogol
Kabupaten Sukoharjo mengucapkan banyak terima kasih kepada bapak ibu bapak saudara
serta tamu undangan.
(9) Atas dukungan atas keridhoannya sehingga saya bisa terpilih menjadi kepala desa di Desa
Manang.
(10) Terima kasih yang tak terhingga, saya ingin mengucapka segera kepada panitia yang telah
menyelenggarakan pemilihan kepala desa.
(11) Pihak pemerintah Desa Manang membutuhkan masukan dan saran dari bapak, ibu, dan
tokoh masyarakat dan siapapun itu yang memiliki masukan, pendapat atau saran yang
membangun untuk kemajan desa kita.
(12) Mulai dari tingkat yang paling bawah, tingkat RT atau juga di tingkat RW, Rukun
Tetangga dan juga Rukun Warga.
(13) Akan kita tindak lanjuti kaitannya dengan masyarakat penerimaan bantuan tersebut.
(14) Salawat serta salam semoga tetap senantiasa tercurahkan kepada junjungan kita Nabiyullah
Muhammad Shallallahu Alaihi Wasallam.
(15) Mulai untuk eee pembangunan atau pemberdayaan masyarakat di Desa Manang ini.
(16) Nah, padahal kegiatan ronda tersebut bisa mewujudkan keamanan dan kenyamanan bagi
warga.
(17) Selanjutnya selain kegiatan ronda di tingkata atau bagian bapak-bapak kita akan walaupun
tahun-tahun sebelumnya sudah.
(18) Selanjutnya di tingkat atau di kegiatan PKK yang tadinya juga ibu-ibu kalau di PKK tinggkat
RW.
Kesalahan berbahasa pada data (7), (8), (9), (10), (11), (12), merupakan kesalahan mubazir kata
atau penggunaan kata yang berlebihan. Penulisan kata camat kecamatan pada data (7), seharusnya cukup
menggunakan kata camat sebagai bentuk penyebutan nama terhormat. Penulisan kata mengucapkan banyak
terima kasih pada data (8), seharusnya cukup menggunakan kata mengucapkan terima kasih sebagai bentuk
kata santun atau pujian. Penulisan kata atas dukungan atas keridhoannya pada data (9), seharusnya cukup
menggunakan kata atas dukungan dan keridhoannya, sebagai bentuk rasa terima kasih. Penulisan kata
terima kasih yang tak terhingga pada data (10), seharusnya cukup menggunakan kata terima kasih, sebagai
bentuk santun atau pujian. Penulisan kata pada kalimat masukan dan saran dari bapak, ibu, dan tokoh
masyarakat dan siapapun itu yang memiliki masukan pada data (11), seharusnya cukup menggunakan kata
“masukan dan saran dari bapak dan ibu, serta tokoh masyrakat”, sebagai bentuk memeberikan rasa
solidaritas antara warga desa. Penulisan kata pada kalimat Mulai dari tingkat yang paling bawah, tingkat
RT atau juga di tingkat RW, Rukun Tetangga dan juga Rukun Warga pada data (12), seharusnya “mulai
dari tingkat RT maupun RW”, sebagai bentuk kalimat keterangan. Penelitian yang setopik dengan
penggunaan kata mubazir dilakukan oleh Astuti et. al (2020) penggunaan kalimat perubahan perubahan
dan perbaikan-perbaikan termasuk kata yang berlebihan.
Penulisan kata tindak lanjuti pada data (13) merupaakan kata tidak baku, seharusnya menggunakan
kata baku yaitu tindak lanjut, sebagai bentuk pertanggungjawaban. Penulisan kata salawat pada data (14)
merupakan kata tidak baku, seharusnya selawat yang merupakan kata baku. Penulisan kata eee pada data
(15) dan penulisan kata nah pada data (16) merupakan kata yang tidak jelas, seharusnya kata eee tidak perlu
digunakan karena bukan kata atau ejaan yang disempurnakan dan juga tidak terdapat di dalam KBBI.
Selanjutnya pada data (10) terima kasih yang tak terhingga, saya ucapkan kepada segera panitia yang telah
menyelenggarakan pemilihan kepala desa. terdapat kalimat yang tidak jelas yang sulit dipahami,
seharusnya “saya..mengucapkan terima..kasih, kepada..panitia yang tellah..menyelenggarakan
pemilihan..kepala desa”, sebagai bentuk pujian dan apresiasi.
Penulisan kesalahan berbahasa pada data (17) dan (18) merupakan kalimat ambigu atau kalimat
yang sulit untuk dipahami dan menimbulkan makna yang sulit dipahami. Penulisan kalimat pada data (17)
seharusnya “pada tahun-tahun sebelumnya kegiatan ronda bapak-bapak sudah terlaksana”. Selanjutnya
pada data (18) seharusnya “selanjutnya kegiatan ibu-ibu PKK di tingkat RW”. Penelitian ini relevn dengan
penelitian yang dilakkan oleh Markhamah & Sabardila (2014) menginformasikan kalimat tidak jelas
merupakan kalimat yang menyebabkan pembaca kesulitan memahami maksud dalam kalimat.
Kesalahan Ejaan
Hortatori | Jurnal Pendidikan Bahasa dan Sastra Indonesia | Vol. 7 No. 2 (2023), 133-140
ISSN 2579-7832 (Cetak) | ISSN 2579-7840 (Elektronik) | Universitas Indraprasta PGRI
138
Bentuk Kesalahan Berbahasa pada Pidato Mahasiswa yang Berperan sebagai Kepala Desa Terpilih
Bentuk kesalahan ejaan juga diitemukan pada penelitiian ini, meliputi kesallahan penulisan tanda baca dan
kata atau tanda hubung. Adapun data yang terdapat dalam wacana pidato tersebut sebagai berikut:
(19) Selanjutnya selain kegiatan ronda di tingkatan atau bagian bapak-bapak kita akan walaupun
tahun-tahun sebelumnya sudah.
(20) Selanjutnya di tingkat atau di kegiatan PKK yang tadinya juga ibu-ibu kalau di PKK tingkat
RW.
Penulisan kesalahan ejaan dalam penelitian ini yaitu terdapat pada data (19) dan (20) yaitu
seharusnya menggunakan tanda koma (,) setelah kata selanjutnya. Penulisan tanda koma yang benar yaitu
pada data (19) “Selanjutnya, selain kegiatan ronda di tingkata atau bagian bapak-bapak kita akan
walaupun tahun-tahun sebelumnya sudah”. Penulisan yang benar pada data (20) yaitu “Selanjutnya, di
tingkat atau di kegiatan PKK yang tadinya juga ibu-ibu kalau di PKK tingkat RW”. Penelitian terdahulu
yang setopik dengan kesalahan penggunaan ejaan dilakukan oleh Casim et. al (2020) terdapat kesalahan
ejaan penggunaan kata depan. Penelitian Rahayu & Sudaryanto (2018) terdapat kesalahan berbahasa
Indonesia yang meliputi kesalahan ejaan. Penelitian Maulida (2021) kesalahan ejaan sering ditemukan
hingga saat ini adalah penulisan partikel.
Simpulan
Hasil dan pembahasan di atas menghasilkan empat kesimpulan pokok, yaitu: (1) Dalam bidang fonologi,
terdapat lima data kesalahan berbahasa, yaitu kesalahan penggunaan huruf atau fonem yang hilang,
kesalahan penulisan atau penggunaan huruf besar atau kapital, dan kesalahan penulisan atau penggunaan
huruf cetak miring; (2) Pada bidang morfologi terdapat satu data yaitu pengulangan kata atau reduplikasi;
(3) Pada bidang sintaksis, terdapat dua belas data kesalahan bahasa, yakni kesalahan penggunaan kata yang
mubazir, kata yang tidak baku, dan kalimat yang tidak jelas; dan (4). Kesalahan dalam bidang ejaan,
terdapat dua data kesalahan yang berkaitan dengan kesalahan tanda baca. Kesalahan berbahasa bisa terjadi
dikarenakan kurangnya pemahaman penutur terhadap kaidah bahasa Indonesia yang baik dan benar.
Kesalahan berbahasa bisa terjadi dikarenakan kurangnya pemahaman penutur terhadap kaidah bahasa
Indonesia yang baik dan benar. Melalui pembelajaran bahasa yang rutin, kompetensi berbahasa seseorang
dapat diperbaiki menjadi lebih baik.Secara keseluruhan, implikasi dari kesimpulan penelitian ini mencakup
perbaikan pendidikan bahasa, peningkatan kesadaran, pengembangan materi pembelajaran, dan upaya
individu untuk meningkatkan kompetensi berbahasa. Dengan langkah-langkah ini, diharapkan kualitas
komunikasi dalam bahasa Indonesia dapat ditingkatkan, baik dalam konteks formal maupun informal.
Ucapan Terima Kasih
Penulis mengucapkan rasa terima kasih kepada dosen pengampu mata kuliah Keterampilan Berbahasa
(Berbicara dan Menulis) di Universitas Muhammadiyah Surakarta, atas bimbingan dan dukungannya dalam
menyelesaikan artikel penelitian ini. Bimbingan dari beliau sangat berarti dan membantu penulis dalam
menghasilkan penelitian ini dengan baik.
Daftar Rujukan
Agustina, Tiya, and Wahyu Oktavia. "Analisis Kesalahan Berbahasa pada Bahan Ajar Kelas Menyimak
Program BIPA IAIN Surakarta." Disastra: Jurnal Pendidikan Bahasa Dan Sastra Indonesia 1.2
(2019): 146-156.
Ariesta, Wiwik, and Atiqa Sabardila. "Kesalahan Berbahasa Bidang Linguistik pada Pidato Mahasiswa
MPBI-UMS yang Berperan sebagai Bupati Terpilih Boyolali." Literasi: Jurnal Bahasa dan Sastra
Indonesia serta Pembelajarannya 5.2 (2021): 345-354.
Arina, Sikana, Nugroho Antoni Mana, and Tahe Agus. "Kesalahan berbahasa tataran fonologi pada pidato
juru bicara penanganan virus COVID-19 Achmad Yurianto." Disastra: Jurnal Pendidikan Bahasa
dan Sastra Indonesia (2021): 74-81.
Busyro, Busyro. "Analisis Kesalahan Kalimat Bahasa Indonesia Pada Teks Pidato Siswa Mts. Ma’arif 16
Nurul Hidayah Banyubang Solokuro Lamongan." Tasyri: Jurnal Tarbiyah-Syariah-Islamiyah 27.1
(2020): 74-83.
Hortatori | Jurnal Pendidikan Bahasa dan Sastra Indonesia | Vol. 7 No. 2 (2023), 133-140
ISSN 2579-7832 (Cetak) | ISSN 2579-7840 (Elektronik) | Url: https://journal.unindra.ac.id/index.php/hortatori/index
Dewi Sartika, Atiqah Sabardila
139
Casim, Yuliani, Y., & Nuraeni, N. S. Analisis Kesalahan Ejaan Berdasarkan Tataran Morfologi pada Media
Iklan di Kota Tasikmalaya. Jurnal Lentera 3.1(2020) 223–229.
Dari, Ulan, and Nyayu Lulu Nadya. "Pemerolehan Bahasa Anak Usia Tiga Tahun Dalam Bidang
Sintaksis." Jurnal Didactique Bahasa Indonesia 3.2 (2022): 67-75.
Diari, Komang Putri Yadnya. "Proses Morfologis Istilah-Istilah Dalam Tajen." Widyacarya: Jurnal
Pendidikan, Agama dan Budaya 2.2 (2019): 85-90.
Fajarwati, Neni Desi. "Kesalahan Siswa dalam Berpidato Bahasa Inggris." Indonesian Journal of Applied
Linguistics Review 2.1 (2017): 47-66.
Fatimah, Fauziah Nurul, et al. "Analisis Kesalahan Berbahasa pada Tuturan Pembawa Acara dan Bintang
Tamu dalam Talk Show Hitam Putih yang Berjudul “Fenomena Kanjeng Dimas”." Parole: Jurnal
Pendidikan Bahasa dan Sastra Indonesia 1.5 (2018): 775-786.
Gani, Saida. "Kajian teoritis struktur internal bahasa (fonologi, morfologi, sintaksis, dan semantik)." A
Jamiy: Jurnal Bahasa dan Sastra Arab 7.1 (2019): 1-20.
Giawa, Kasihani. "Analisis Kesalahan Berbahasa dalam Membaca Teks Pidato Oleh Siswa Kelas VIII SMP
Negeri 1 LÖLÖWA’U." FAGURU: Jurnal Ilmiah Mahasiswa Keguruan 1.2 (2022): 317-326.
Hasanudin, Cahyo. "Analisis Kesalahan Berbahasa Pada Penulisan Media Luar Ruang Di Kabupaten
bojonegoro." Jurnal Pendidikan Bahasa dan Sastra UPI 17.1 (2017): 120-129.
Johan, Gio Mohamad. "Kesalahan Fonologis dalam Proses Diskusi Siswa Sekolah Dasar." Jurnal
Metamorfosa 6.2 (2018): 123-133.
Kim, Soo-Jin. "A comparison of phonological error patterns in the single word and spontaneous speech of
children with speech sound disorders." Phonetics and Speech Sciences 7.3 (2015): 165-173.
Kridalaksana. Kamus Linguistik. Jakarta: Gramedia Pustaka Utama. 2008.
Kundharu, Saddhono, and Slamet St Y. "Pembelajaran keterampilan berbahasa Indonesia." Yogyakarta:
Graha Ilmu (2014).
Majid, Abdul. Pengertian Analisis. Bandung: Remaja Rosdakarya.2013.
Maulida, U. (2021). Kesalahan Berbahasa Tataran Ejaan, Morfologi, dan Sintaksis Skripsi Mahasiswa
Program Studi PGMI Binamadani. Dirasah Jurnal Pemikiran & Pendidikan Dasar Islam 4.1
(2021) 24–34. https://doi.org/https://doi.org/10.5 1476/dirasah.v4i1.220
Markhamah, & Sabardila, A. Analisis Kesalahan & Karakteristik Bentuk Pasif. Surakarta: Muhammadiyah
University Press. 2014.
Mustofa, Hendi, and Lina Dwi Safitri. Analisis Kesalahan Berbahasa Bidang Semantik dalam Pidato
Menteri Pendidikan dan Kebudayaan.2021.
Nisak, Khoirun, and Purwati Anggraini. “Kritik Sosial Dalam Novel ‘Anak-Anak Tukang’ Karya Baby
Ahnan.” Alinea: Jurnal Bahasa, Sastra, Dan Pengajaran 9.2 (2020): 146,
https://doi.org/10.35194/alinea.v9i2.990.
Sabardila, Atiqa. Analisis Kesalahan dan Karakteristik Bentuk Pasif. Muhammadiyah University Press,
2014.
Qoyyimah, Atika Lisamawati Nur, and Atiqa Sabardila. "Bentuk Kesalahan Berbahasa dalam Pidato
Mahasiswa Yang Memerankan Diri Sebagai Bupati Terpilih Kabupaten Blora." Literasi: Jurnal
Bahasa dan Sastra Indonesia serta Pembelajarannya 5.2 (2021): 173-186.
Prihantoro, Syukur. "Analisis Kesalahan Bahasa Pada Taksonomi Linguistik Dalam Penulisan Insya’." Al
Mahāra: Jurnal Pendidikan Bahasa Arab 5.1 (2019): 41-62.
Puspitasari, Ai Nita, et al. "Analisis Kesalahan Bahasa dalam Jurnal Auto Tech 2019 Universitas
Muhammadyah Purworejo." ALFABETA: Jurnal Bahasa, Sastra, dan Pembelajarannya 3.2
(2020): 35-40.
Putri, Haipa Novia. “Kemampuan Berpidato Siswa Kelas X SMA Negeri 3 KotabumiTahun Ajaran
2016/2017." (2017).
Rahayu, A., & Sudaryanto. Kesalahan Ejaan, Diksi, dan Morfologi dalam Karangan Deskripsi Mahasiswa
Asal Tiongkok. Diglosia: Jurnal Pendidikan, Kebahasaan, Dan Kesusastraan Indonesia 2.1
(2018). 42– 49. http://eprints.uad.ac.id/14772/
Sintia, Mila, I. Nyoman Sudiana, and I. Gede Nurjaya. "Analisis Kesalahan Morfologi Pada Tuturan
Siswasmp N 3 Banjar." Jurnal Pendidikan Bahasa Dan Sastra Indonesia Undiksha 9.2 (2019).
Saputri, Kurnia. "Analisis Kesalahan Morfologi pada Pidato Presiden Joko Widodo dalam Rangka
Pelantikan Presiden dan Wakil Presiden Terpilih Periode 2019-2024." Jurnal Skripta 5.2 (2019).
Hortatori | Jurnal Pendidikan Bahasa dan Sastra Indonesia | Vol. 7 No. 2 (2023), 133-140
ISSN 2579-7832 (Cetak) | ISSN 2579-7840 (Elektronik) | Universitas Indraprasta PGRI
140
Bentuk Kesalahan Berbahasa pada Pidato Mahasiswa yang Berperan sebagai Kepala Desa Terpilih
Setyawati, Nanik. Analisis Kesalahan Berbahasa Indonesia Teori dan Praktik. Surakarta: Yuma
Pustaka.2010.
Sudaryanto. Metode dan Aneka Teknik Analisis Bahasa: Pengantar Penelitian Wahana Kebudayaan secara
Linguistis. Yogyakarta: Sanata Dharma University Press.2013.
Sugiyono. Metode Penelitian Pendidikan: Pendekatan Kuantitatif, Kualitatif, dan R&D. Bandung:
Alfabeta.2012.
Sudjana, Nana. Pengertian Analisis. Bandung: Remaja Rosdakarya.2016.
Suhardi. Pengantar Linguistik Umum. Jokjakarta: Ar-Ruzz Media, 2013.
Syafi'i, Bahaudin Alfiansyah, and Ira Khoirun Niha. "Analisis kesalahan morfologi dalam penulisan
makalah mahasiswa Hukum Ekonomi Syariah IAIN Surakarta." Jurnal Penelitian Humaniora 22.1
(2021): 14-29.
Thoyib, Thoyib, and Hasanatul Hamidah. "Interferensi Fonologis Bahasa Arab “Analisis Kontrastif Fonem
Bahasa Arab Terhadap Fonem Bahasa Indonesia Pada Mahasiswa Universitas Al Azhar Bukan
Jurusan Sastra Arab”." Jurnal Al-Azhar Indonesia Seri Humaniora 4.2 (2018): 63-71.
Mustofa, Hendi, and Lina Dwi Safitri. Analisis Kesalahan Berbahasa Bidang Semantik dalam Pidato
Menteri Pendidikan dan Kebudayaan.2021.
Hortatori | Jurnal Pendidikan Bahasa dan Sastra Indonesia | Vol. 7 No. 2 (2023), 133-140
ISSN 2579-7832 (Cetak) | ISSN 2579-7840 (Elektronik) | Url: https://journal.unindra.ac.id/index.php/hortatori/index
|
|
https://openalex.org/W2091882320
|
https://gfzpublic.gfz-potsdam.de/pubman/item/item_247564_3/component/file_247563/21092.pdf
|
English
| null |
Numerical simulations of CO2 arrival times and reservoir pressure coincide with observations from the Ketzin pilot site, Germany
|
Environmental earth sciences
| 2,013
|
cc-by
| 6,464
|
Numerical simulations of CO2 arrival times and reservoir
pressure coincide with observations from the Ketzin pilot site,
Germany Thomas Kempka • Michael Ku¨hn Received: 15 November 2012 / Accepted: 20 June 2013
The Author(s) 2013. This article is published with open access at Springerlink.com Abstract
Numerical simulations of CO2 migration for
the period June 2008–December 2011 were performed
based on a unique data set including a recently revised
static geological 3D model of the reservoir formation of the
Ketzin pilot site in Brandenburg, Germany. We applied the
industrial standard ECLIPSE 100 and scientific TOUGH2-
MP simulators for this task and implemented a workflow to
allow for integration of complex model geometries from
the Petrel software package into TOUGH2-MP. Definition
of a near- and a far-field well area allowed us to apply
suitable permeability modifiers, and thus to successfully
match simulation results with pressure measurements and
arrival times in observation wells. Coincidence was
achieved for CO2 arrival times with deviations in the range
of 5.5–15 % and pressure values in the injection well CO2
Ktzi 201/2007 and the observation well CO2 Ktzi 202/2007
with even smaller deviations. It is shown that the integra-
tion of unique operational and observation data in the
workflow improves the setup of the geological model. Within such an iteration loop model uncertainties are
reduced and enable advanced predictions for future reser-
voir behaviour with regard to pressure development and
CO2 plume migration in the storage formation at the Ketzin
pilot site supporting the implementation of monitoring
campaigns and guiding site operation. Environ Earth Sci
DOI 10.1007/s12665-013-2614-6 Environ Earth Sci
DOI 10.1007/s12665-013-2614-6 SPECIAL ISSUE T. Kempka (&) M. Ku¨hn
Section 5.3 - Hydrogeology, GFZ German Research Centre for
Geosciences, Telegrafenberg, 14473 Potsdam, Germany
e-mail: kempka@gfz-potsdam.de
M. Ku¨hn
e-mail: mkuehn@gfz-potsdam.de Introduction The Ketzin pilot site located in the State of Brandenburg,
Germany, was the first European on-shore site for CO2
storage in a saline aquifer and is now operated for[4 years
(Martens et al. 2012; Wu¨rdemann et al. 2010). So far, three
wells (CO2 Ktzi 201/2007 for injection and observation as
well as CO2 Ktzi 200/2007 and CO2 Ktzi 202/2007 for
observation) have been drilled down to the base of the
Stuttgart Formation in 800 m depth in the Keuper. The
target formation for CO2 injection is at a depth of about
630–650 m at the injection well CO2 Ktzi 201/2007. CO2
has been injected since June 2008 at an average rate of
about 0.5 kg/s based on an injection schedule considering
different monitoring campaigns, well tests and operational
revisions. Operational data for the Ketzin pilot site were
recently published by Mo¨ller et al. (2012) covering all
relevant data for simulations from the start of injection up
to the end of the year 2011. The scientific novelty elaborated in the study presented
here is based on a wide range of geological and monitoring
data from the Ketzin pilot site. Compared to other locations
around the world, breadth and depth of the measurements
and longevity of the recorded time series are unique as
shown by Michael et al. (2010) and references therein. This
enables us to parameterise with great detail our static
geological and dynamic flow models (e.g. Norden and
Frykman 2013). Characteristic for that beside others is the
permanent bottom hole pressure measurement and moni-
toring. From that perspective the available data set is
unique because only the pilot sites like Ketzin, Frio, Na-
gaoka and Otway do have bottom hole pressure data
(Michael et al. 2010). The larger demonstration projects of
Sleipner or In Salah are lacking that. The pilot sites Frio
and Nagaoka are orders of magnitude smaller compared to Keywords
Numerical modelling CO2 storage Ketzin
pilot site ECLIPSE TOUGH2 Keywords
Numerical modelling CO2 storage Ketzin
pilot site ECLIPSE TOUGH2 M. Ku¨hn
e-mail: mkuehn@gfz-potsdam.de 12 3 Environ Earth Sci Ketzin with regard to the amount of injected CO2. Otway is
of the same size, however, it is a pilot for CO2 storage in a
depleted gas field. Introduction It is only Ketzin which can provide
bottom hole pressure data to be used for dynamic model
matching for a time-span of [4 years (Mo¨ller et al. 2012)
and a significant amount of injected CO2 into a saline
aquifer (Martens et al. 2012). Taking into account the high
complexity of the geological setting of the fluvial Stuttgart
Formation and the monitoring data density (especially
related to pressure profiling) a sufficiently good match of
bottom hole pressure in acceptable agreement with CO2
arrival times in the observation wells offers an excellent,
unique and new opportunity to validate the numerical
simulation tools currently available. Frykman (2013). The latest geological model revision
(Kempka et al. 2013b) is employed for the reservoir sim-
ulations presented in this study. We expected now a better match between observations
and simulations based on the revised static geological 3D
model independent of the application of two different
simulators. We wanted to show that integration of new
observation data improves the quality of a static geological
model significantly to better match observation data with
each iteration. The presented work is a continuation from
Kempka et al. (2010), whereas dynamic flow coupled with
geochemical simulations accounting for CO2 trapping
mechanisms are discussed in Klein et al. (2013), Kempka
et al. (2013a) and Fischer et al. (2013). Here, the industrial
standard simulator ECLIPSE 100 (Schlumberger 2009) and
the scientific simulator TOUGH2-MP/ECO2N (Zhang
et al. 2008; Pruess 2005) were applied for this unique
dynamic flow simulation study. An extended simulator
intercomparison study using the Ketzin pilot site static
geological model and monitoring data is presented in Ke-
mpka et al. (2013b). Kempka et al. (2010) discussed predictive models for
CO2 injection at the Ketzin site. These simulations were
based on the static geological 3D model available at that
time which did not include the interpretation of the 3D
seismic repeat from late October 2009. All available geo-
physical and geological information is used for recurring
updates of the static model. In 2010, Kempka et al. determined that the simulated CO2 arrival at the first
observation well (CO2 Ktzi 200/2007) can be matched
with deviations of 21–33 % in the predictive case and
8.1–17.7 % applying the actual injection regime neglecting
the pressure development in the reservoir. It was therefore
concluded that knowing the injection regime is of utmost
importance to be able to predict the CO2 migration in the
reservoir more accurately. Introduction The arrival time of the CO2 at
the second observation well (CO2 Ktzi 202/2007) could not
be matched by any of the applied codes, indicating that the
geological model did not represent the actual geological
conditions at that time. Hence, complete coincidence
between monitoring and modelling with the previous static
geological 3D model could not be gained with regard to
reservoir pressure and arrival times. Considering these
simulation results, Lengler et al. (2010) employed a Monte
Carlo analysis to assess heterogeneity in porosity and
permeability by means of representing a potential driver for
the late CO2 arrival observed at the CO2 Ktzi 202/2007
well. They demonstrated that heterogeneity is very unlikely
responsible for the deviation in CO2 arrival time at the
second observation well. Through these findings and sup-
ported by updates in monitoring data, the geological model
was revised by implementation of a facies-based hetero-
geneity in the CO2 Ktzi 202/2007 near-well area consisting
of floodplain and sand channel facies with low and high
permeabilities, respectively, as discussed in Kempka et al. (2013b). 123 Model geometry and discretisation 1 3D view of the reservoir model of the Ketzin anticline (5 9 5 km 9 about 72 m) with three different la
zones and seven major faults at the top of the anticline (vertically exaggerated by factor 5) thickness with 1 m elements and Zone C: about 36 m
thickness with 3 m elements) was up-scaled to 2 m ele-
ments in Zone B and 6 m elements in Zone C, while ver-
tical discretisation in Zone A was not modified to account
for buoyancy effects of the multi-phase system by a suf-
ficient vertical resolution of the uppermost layers (Fig. 1). The resulting reservoir model consists of 648,420 active
elements discretised by 107 elements in i-, 101 elements in
j- and 60 elements in k-direction. corner coordinates, centres and volumes determined
according to the approach of Grandy (1997). Additional
information such as, the connection distance, volume,
interface area to other lateral and horizontal neigh-
bouring
elements
are
directly
written
into
the
TOUGH2-MP output file. Further pre-processing tools
are applied to define initial and boundary conditions. Parameterisation of the ECLIPSE and TOUGH2 models
was done as discussed in Kempka et al. (2010) considering
the following revisions: •
ranges of effective porosities and initial permeabilities
as presented by Norden and Frykman (2013) were
updated based on the latest observations at the Ketzin
pilot site (Kempka et al. 2013b); Model geometry and discretisation Model geometry and discretisation The implementation of the initial static geological 3D
model of the Ketzin anticline is described in detail by
Norden and Frykman (2013). Meanwhile, observation data
available for a period until the end of 2011 has been
integrated
into
the
geological
model
(3D
seismic
(re-)interpretation, electrical resistivity tomography and
well logging) in addition to updated operational data
(pressures and injection rates). The geological model has a
lateral size of 5 9 5 km and a thickness of about 72 m also
involving seven major faults with the respective offsets at
the top of the anticline. In the first step, we up-scaled the
reservoir properties relevant for dynamic simulations
(effective porosity, permeability and facies description)
from the initially very fine geological grid of 5 9 5 m
lateral size to the grid of our reservoir model illustrated in
Fig. 1 using the Petrel 2010 software package (Schlum-
berger 2010). For that purpose, we defined three lateral
zones of different discretisation: a near-well zone of
150 9 200 m and 5 9 5 m element size (Fig. 2); a zone
considering the size of the expected CO2 plume for the
envisaged simulation time of 3.5 years of 3.5 9 1.5 km
and 50 9 50 m element size (Fig. 3); and an outer zone
(5 9 5 km) with 100 9 100 m element size (Fig. 3). The
initial vertical discretisation applied in the revised geo-
logical model consists of three zones (Zone A: about 24 m
thickness with 0.5 m elements, Zone B: about 12 m Updates of monitoring data obtained by 3D seismic
(re-)interpretation, electrical resistivity tomography and
well logging allowed for a revision of the initial static
geological model which is presented by Norden and 123 123 Environ Earth Sci Fig. 1 3D view of the reservoir model of the Ketzin anticline (5 9 5 km 9 about 72 m) with three different lateral and vertical discretisation
zones and seven major faults at the top of the anticline (vertically exaggerated by factor 5) Fig. 1 3D view of the reservoir model of the Ketzin anticline (5 9 5 km 9 about 72 m) with three different lateral and vertical discretisation
zones and seven major faults at the top of the anticline (vertically exaggerated by factor 5) Fig. Fig. 2 Planar view of near-well
discretisation applied in the
reservoir model with the CO2
Ktzi 201/2007 injection as well
as the CO2 Ktzi 200/2007 and
CO2 Ktzi 202/2007 observation
wells in about 50 and 112 m
distance from the CO2 Ktzi
201/2007 well, respectively.
Near-well lateral element size is
5 9 5 m Model parameterisation We developed a workflow allowing for a conversion of
Petrel data sets into the TOUGH2-MP/ECO2N input
data format that was applied in order to ensure identical
implementation in terms of geometry, discretisation and
properties of the reservoir model in both simulators
applied. This
workflow
comprises
multiple
steps
beginning with reading in of the initial reservoir sim-
ulation structured hexahedron grid constructed using an
arbitrary pre-processing software package (e.g. Sch-
lumberger Information Services Petrel) and its storage
in an adapted data structure representing element ids, •
initial permeability anisotropy was determined as Kv/
Kh = 1/3, while the final permeability anisotropy was
determined by matching observation and simulation
data; •
initial pressure is 62 bar at a depth of 639.5 m in the
CO2 Ktzi 201/2007 well and temperature is considered
to be constant at 34 C in the entire reservoir (unpub-
lished data); •
brine salinity is 0.22 kg NaCl equivalents per kg brine; 12 Environ Earth Sci Fig. 2 Planar view of near-well
discretisation applied in the
reservoir model with the CO2
Ktzi 201/2007 injection as well
as the CO2 Ktzi 200/2007 and
CO2 Ktzi 202/2007 observation
wells in about 50 and 112 m
distance from the CO2 Ktzi
201/2007 well, respectively. Near-well lateral element size is
5 9 5 m •
the skin factor is neglected as it does not notably
influence the simulation results; •
the skin factor is neglected as it does not notably
influence the simulation results; Consequently, we defined all faults to be impermeable in
vertical direction. Integrity of the upper (Weser Formation)
and lower caprocks (Grabfeld Formation) are considered
by no-flow top and bottom boundary conditions, while an
infinite aquifer is accounted for by a (pore) volume mul-
tiplication factor of 10,000 at the lateral boundary elements
to avoid boundary pressure effects. Consequently, we defined all faults to be impermeable in
vertical direction. Integrity of the upper (Weser Formation)
and lower caprocks (Grabfeld Formation) are considered
by no-flow top and bottom boundary conditions, while an
infinite aquifer is accounted for by a (pore) volume mul-
tiplication factor of 10,000 at the lateral boundary elements
to avoid boundary pressure effects. •
residual water saturation is Swr = 0.15 and residual gas
saturation Sgr = 0.05 as determined in laboratory
experiments (unpublished data); •
relative permeabilities as well as capillary pressure
were derived from flow-through experiments on core
samples from the Stuttgart Formation (unpublished
data). Model parameterisation The input data for both simulators are illustrated
in Fig. 4. The well implementation differs in both simulators as
TOUGH2 does not come with a wellbore module, while
ECLIPSE does. Hence, a simplified approach was chosen
in the TOUGH2 simulations by assigning a high porosity of
0.99 and permeability of 1,000 mD to the cell column
representing the well instead of the data derived from up-
scaling. ECLIPSE uses the original element porosities and
permeabilities of the element column penetrated by the
well, since the internal wellbore flow module can be
applied. Open sections of the three wells (filter screens) at
reservoir depth could be implemented identically in both
models according to operational data (unpublished data). The seven major faults at the top of the Ketzin anticline
were not implemented in the previous model (Kempka
et al. 2010). As there is no indication of low horizontal
fault permeability based on hydraulic tests undertaken
(Wiese et al. 2010) and unpublished operational data of the
gas storage operation, all seven faults implemented in the
reservoir model were assumed to be horizontally conduc-
tive for fluid flow. According to permanent pressure and
isotope monitoring in an indicator horizon located in the
Exter Formation (about 180 m above the Stuttgart For-
mation), regional communication between that indicator
horizon and aquifers in the Jurassic or the Stuttgart For-
mation may exist (Wiese et al. 2013). However, there is no
evidence of CO2 or brine migration from the Stuttgart
Formation via the faults present at the anticline top. Fig. 4 Relative permeabilities
for the wetting (krw) and non-
wetting phases (krn) as well as
capillary pressure determined
by laboratory experiments and
smoothened in order to assure
convergence behaviour in the
applied numerical simulators Pressure correction and injection rate For that purpose, a density-based approach
was chosen by discretising the vertical well section (89.5 m
to the reference depth of 639.5 m) into vertical elements of
arbitrary size and cumulating the pressure-dependent CO2
density (and N2 density during injection stops in the first
year of operation) as a function of pressure and temperature
conditions in the well by application of the equation of
state for gas mixtures developed by Kunz et al. (2007). A
vertical well discretisation with 100 elements was found to
provide sufficient accuracy based on a sensitivity analysis. Pressure loss was determined to be \0.3 bar in the CO2
Ktzi 201/2007 injection well, and thus is neglected in the
pressure correction calculations. The measured pressure at
550 m and the pressure corrected to 639.5 m depth (ref-
erence depth for reservoir simulations) as well as the
cumulative injected CO2 mass until the end of 2011 are
shown in Fig. 5. in i-, j- and k-directions as listed in Table 1 to avoid an
increase of simulated pressure not corresponding with the
observations as of March 2010. Table 2 shows the observed arrival times of gaseous
CO2 in both observation wells determined using a gas
membrane sensor (Zimmer et al. 2011) as well as the
simulated arrival times of gaseous and dissolved CO2. In
contrast to the results described by Kempka et al. (2010),
the geological model is now able to provide a significantly
better representation of the monitored CO2 behaviour. This
can be deduced from the agreement of simulated CO2
arrival times at the CO2 Ktzi 200/2007 well with devia-
tions of 6.4 % (ECLIPSE) and 11.1 % (TOUGH2) related
to the observed data, as well as the match for the CO2 Ktzi
202/2007 well with deviations of 6.4 % (ECLIPSE) and
15.2 % (TOUGH2). Arrival times of dissolved CO2 (not
measured at the observation wells) are in agreement
between both simulators for both observation wells. The matching process was undertaken considering the
pressure corrected to the reference depth in the CO2 Ktzi
201/2007 well and measured at 623.8 m in the CO2 Ktzi
202/2007 observation well (installed for about 1.5 years). The results illustrated in Fig. Pressure correction and injection rate CO2 injection rates as well as temperature and pressure
measured at 550 m depth in the CO2 Ktzi 201/2007 well
from the start of injection until the end of 2011 are derived 123 123 Environ Earth Sci Fig. 3 Planar view of the different discretisation zones accounting for the near-well area (150 9 200 m), the expected CO2 plume size at the
envisaged simulation time (3.5 9 1.5 km) and the outer zone (5 9 5 km) Fig. 3 Planar view of the different discretisation zones accounting for the near-well area (150 9 200 m), the expected CO2 plume size at the
envisaged simulation time (3.5 9 1.5 km) and the outer zone (5 9 5 km) Fig. 4 Relative permeabilities
for the wetting (krw) and non-
wetting phases (krn) as well as
capillary pressure determined
by laboratory experiments and
smoothened in order to assure
convergence behaviour in the
applied numerical simulators
123 Fig. 4 Relative permeabilities
for the wetting (krw) and non-
wetting phases (krn) as well as
capillary pressure determined
by laboratory experiments and
smoothened in order to assure
convergence behaviour in the
applied numerical simulators Fig. 4 Relative permeabilities
for the wetting (krw) and non-
wetting phases (krn) as well as
capillary pressure determined
by laboratory experiments and
smoothened in order to assure
convergence behaviour in the
applied numerical simulators 12 3 Environ Earth Sci Fig. 5 Pressure measured in the injection well CO2 Ktzi 201/2007 at 550 m and cumulative injected CO2 mass after Mo¨ller et al. (2012) as well
as pressure corrected to the reference depth of 639.5 m Fig. 5 Pressure measured in the injection well CO2 Ktzi 201/2007 at 550 m and cumulative injected CO2 mass after Mo¨ller et al. (2012) as well
as pressure corrected to the reference depth of 639.5 m n the injection well CO2 Ktzi 201/2007 at 550 m and cumulative injected CO2 mass after Mo¨ller et al. (2012) as well
reference depth of 639.5 m from Mo¨ller et al. (2012). Since the top of the reservoir is
located about 80 m below the pressure and temperature
sensor installed in the CO2 Ktzi 201/2007 well, a pressure
correction to reservoir depth had to be applied to allow us
for a comparison of simulation results and observed res-
ervoir pressure. Pressure correction and injection rate 6 show agreement at the CO2
Ktzi 201/2007 well for both simulators in the first 4 months
of injection and a slight underestimation of pressure (by
about 0.7 bars) by both simulators from month November
2008 to June 2009, whereas ECLIPSE results show a good Results The CO2 Ktzi 202/2007 pressure match is as well in
agreement with the observations for both simulator results
as well as for the comparison between both simulators
against each other where monitoring data are not available
(e.g. before May 2010). Figure 7 shows the balance of injected, gaseous and
dissolved CO2 for the entire simulation time. Both simu-
lation results are in agreement with each other. Peaks
occurring in the ECLIPSE data are not present in the
TOUGH2 data due to a limited amount of output data. However, the intermediate values coincide. Dissolution of
CO2 is slightly higher in the ECLIPSE model at the end of
the simulated time. Results Simulation of the pressure at reference depth in the CO2
Ktzi 201/2007 injection well as well as CO2 arrival times in
the observation wells CO2 Ktzi 200/2007 and CO2 Ktzi
202/2007 lead to results in good agreement with our
observations applying permeability modifiers in two lateral
areas of the reservoir model. For that purpose, a near-well
area was defined for the range of i = 30–64 and j = 49–79
(origin in the upper left corner) and a far-well area for all
elements not included in the near-well area considering the
entire model thickness. Permeability multipliers were
applied in both models to assign the revised permeabilities Table 1 Permeability multipliers applied at the near- and far-well
areas as a result of the matching process for reservoir pressure and
CO2 arrival times Table 1 Permeability multipliers applied at the near- and far-well
areas as a result of the matching process for reservoir pressure and
CO2 arrival times Table 1 Permeability multipliers applied at the near and far well
areas as a result of the matching process for reservoir pressure and
CO2 arrival times
Permeability multiplier
Near-well area
Far-well area
i-direction
0.34
0.38
j-direction
0.07
0.10
k-direction
0.07
0.10 123 Environ Earth Sci Table 2 Observed and simulated arrival times of gaseous and dissolved CO2 in the observation wells CO2 Ktzi 200/2007 and CO2 Ktzi
202/2007
Observation
well
Phase
Observed
(days)
Simulated
TOUGH2 (days)
Simulated
ECLIPSE (days)
Deviation
TOUGH2 (%)
Deviation
ECLIPSE (%)
CO2 Ktzi 200/2007
Dissolved CO2
–
10.6
11.0
–
–
Gaseous CO2
21.7
19.3
20.3
11.1
6.4
CO2 Ktzi 202/2007
Dissolved CO2
–
221.4
225.0
–
–
Gaseous CO2
271
229.8
256.0
15.2
5.5
Fig. 6 Observed and matched pressure in the injection well CO2 Ktzi 201/2007 (permanent pressure monitoring) and the second observation
well CO2 Ktzi 202/2007 (pressure sensor installed from May 2010 to October 2011) Table 2 Observed and simulated arrival times of gaseous and dissolved CO2 in the observation wells CO2 Ktzi 200/2007 and CO2 Ktzi
202/2007 val times of gaseous and dissolved CO2 in the observation wells CO2 Ktzi 200/2007 and CO2 Ktzi Fig. 6 Observed and matched pressure in the injection well CO2 Ktzi 201/2007 (permanent pressure monitoring) and the second observation
well CO2 Ktzi 202/2007 (pressure sensor installed from May 2010 to October 2011) Fig. Results 6 Observed and matched pressure in the injection well CO2 Ktzi 201/2007 (permanent pressure monitoring) and the second observation
well CO2 Ktzi 202/2007 (pressure sensor installed from May 2010 to October 2011) match after February 2009. In the following months, the
TOUGH2 results match with the observed data, while
ECLIPSE tends to overestimate the pressure peaks result-
ing from injection at maximum rates (about 2.000 kg/s) by
up to 0.8 bars. However, subsequently declining pressure is
matched by the ECLIPSE simulator. Interference tests
(alternating injection and shutdown) conducted from Sep-
tember to October 2010 are depicted by both simulators as
well as different injection stops performed for monitoring
campaigns and operational revisions during the entire time-
span. The CO2 Ktzi 202/2007 pressure match is as well in
agreement with the observations for both simulator results
as well as for the comparison between both simulators
against each other where monitoring data are not available
(e.g. before May 2010). 2011. The simulations are in agreement at the first time
steps with low deviations, e.g. the CO2 plume calculated
with TOUGH2 extends by 10–20 m earlier in northwest
direction, while TOUGH2 results show a rather concen-
trated thickness around the injection well CO2 Ktzi
201/2007 for the two later time steps, whereby the overall
CO2 plume shape is almost identical. In general, maximum
CO2 plume thickness of up to 25.5 m is simulated at the
CO2 Ktzi 201/2007 well and the lateral CO2 plume size at
the end of July 2011 is about 1,200 9 1,000 m. Figure 9
indicates that the saturation of the CO2-rich phase calcu-
lated by both simulators at 31 July 2011 is almost identical
with regard to shape and thickness. However, the well
implementation in the TOUGH2 model leads to higher CO2
saturations in the well elements compared to those calcu-
lated by ECLIPSE. match after February 2009. In the following months, the
TOUGH2 results match with the observed data, while
ECLIPSE tends to overestimate the pressure peaks result-
ing from injection at maximum rates (about 2.000 kg/s) by
up to 0.8 bars. However, subsequently declining pressure is
matched by the ECLIPSE simulator. Interference tests
(alternating injection and shutdown) conducted from Sep-
tember to October 2010 are depicted by both simulators as
well as different injection stops performed for monitoring
campaigns and operational revisions during the entire time-
span. Discussion and conclusions Within this manuscript, we outlined our most recent
matching activities for the Ketzin pilot site based on a
revised version of the initial static geological 3D model
involving permeability fitting based on field observations
of CO2 arrival and pressure monitoring with its general Figure 8 shows CO2 plume thickness isolines at 26
November 2008, 04 June 2009, 13 June 2010 and 31 July 12 3 Environ Earth Sci Fig. 7
Balance of total injected CO2, free gas phase and dissolved CO2 for both simulators applied Fig. 7
Balance of total injected CO2, free gas phase and dissolved CO2 for both simulators applied (well location in ECLIPSE) to the element interface, as two
well elements are always affected in this assessment. This
explanation appears reasonable for both arrival times when
considering, e.g. the CO2 arrival at the CO2 Ktzi 202/2007
well with 258 days calculated by ECLIPSE and 229.8 days
by TOUGH2, whereby an average flow velocity of 0.43 m/
day is achieved. This would roughly sum up to about
240 days arrival time for the TOUGH2 simulation, even
though the decrease of flow velocity due to the radial
propagation of the CO2 plume is neglected for this
estimation. implementation discussed by Norden and Frykman (2013). For the numerical simulations we applied two different
computer codes for comparison using a complex hetero-
geneous geological model taking into account all known
reservoir properties: the industrial standard simulator
ELCIPSE 100 and the scientific simulator TOUGH2-MP/
ECO2N. A workflow for the conversion of complex Petrel
models into TOUGH2 simulation models was developed
and allows us now a fast integration of complex geometries
like fault systems with offsets into the TOUGH2 simulator. Operational data of the Ketzin pilot site were integrated to
parameterise the reservoir model. The simulated pressure coincides with the reservoir
pressure observed showing few deviations only. We expect
that these are mainly caused by the different well imple-
mentations in both simulators. This assumption is under-
lined by the agreement of pressure data observed in the
CO2 Ktzi 202/2007 well and the related pressures calcu-
lated by both simulators (Fig. 6). Definition of a near- and far-field well area allowed us to
assign different permeability modifiers in i-, j-, k-directions
in order to achieve a successful match between observed
and simulated data for the geological model (Table 1). Discussion and conclusions A
factor of about four in lateral anisotropy represented by the
permeability multipliers which were determined during the
matching process is in agreement with the hydraulic tests
undertaken in advance of CO2 injection in 2008 as dis-
cussed by Wiese et al. (2010). CO2 balance in terms of gaseous and dissolved CO2 is
comparable over the entire simulation time for both sim-
ulators (Fig. 7) and spatial distribution of the CO2 plume
and its thickness are also almost identical for both simu-
lators (Fig. 8), whereby slight deviations (concentrated
CO2 plume thickness in the near-well area of CO2 Ktzi
201/2007 in the TOUGH2 results) are again expected to
result from the different well implementations in both
simulators. In the TOUGH2 model more CO2 can be
present in the CO2 Ktzi 201/2007 well elements due to the
chosen porosity of 0.99. Simulation results are in agreement with observed CO2
arrival times, whereas TOUGH2 simulation results tend to
have deviations of 11 % (CO2 Ktzi 200/2007) to 15 %
(CO2 Ktzi 202/2007) compared to those of ECLIPSE with
5.5 % (CO2 Ktzi 202/2007) to 6.4 % (CO2 Ktzi 200/2007)
as presented in Table 2. Considering the marginal varia-
tions of arrival time of dissolved CO2 (Table 2) and the
coincidence of CO2 plume thickness and spatial extent over
time (Fig. 8), we expect that the well implementation in the
TOUGH2 reservoir model is responsible for the deviations
encountered. Here,
relatively
high
permeabilities
of
1,000 mD were assigned to the well elements of 5 9 5 m
lateral size leading to a decrease in the flow path by 5 m in
total according to the double distance of the cell centre Computational
times
of
the
TOUGH2-MP/ECO2N
simulator were about 24 times higher for the current model
compared to those of the ECLIPSE 2009.2 simulator using
the same computer architecture. However, the application
of a high performance computing cluster with 256 cores
resulted in faster simulations compared to a sequential 123 Environ Earth Sci ( i
l
) ECLIPSE
h
th t t l
t ti
l
id
i
it
i
d t
bt i
d
t th
K t i
il t
Fig. 8 CO2 plume thickness in meter plots on 26 November 2008
(first row), 04 June 2009 (second row), 13 June 2010 (third row) and
31 July 2011 (fourth row), whereas ECLIPSE results are plotted left
and TOUGH2 results right. Discussion and conclusions The size of the squares in the background
is 200 9 200 m Fig. 8 CO2 plume thickness in meter plots on 26 November 2008
(first row), 04 June 2009 (second row), 13 June 2010 (third row) and
31 July 2011 (fourth row), whereas ECLIPSE results are plotted left and TOUGH2 results right. The size of the squares in the background
is 200 9 200 m (single core) ECLIPSE run, where the total computational
time was about 2 days. wide unique monitoring data obtained at the Ketzin pilot
site. We emphasize here that integration of all available
operational and monitoring data improve the static geo-
logical model within the recurring update resulting in In summary, we were able to achieve satisfying
agreement between our simulation results with the world- 12 3 Environ Earth Sci Fig. 9 Cross-section of the CO2-rich phase saturation at the CO2
Ktzi 201/2007 injection well (indicated as black line) at 31 July 2011
calculated with ECLIPSE (left) and TOUGH2/ECO2N (right). Near-
well lateral element size is 5 and 50 m thereafter, while element
thickness in the uppermost zone corresponds to 0.5 m well lateral element size is 5 and 50 m thereafter, while element
thickness in the uppermost zone corresponds to 0.5 m he CO2-rich phase saturation at the CO2
ell (indicated as black line) at 31 July 2011
(left) and TOUGH2/ECO2N (right). Near-
well lateral element size is 5 and 50 m thereafter, while element
thickness in the uppermost zone corresponds to 0.5 m well lateral element size is 5 and 50 m thereafter, while element
thickness in the uppermost zone corresponds to 0.5 m Fig. 9 Cross-section of the CO2-rich phase saturation at the CO2
Ktzi 201/2007 injection well (indicated as black line) at 31 July 2011
calculated with ECLIPSE (left) and TOUGH2/ECO2N (right). Near- Acknowledgments
The authors gratefully acknowledge the funding
support from the German Federal Ministry of Education and Research
(Grant 03G0760A-F) and the industry partners VNG, Vattenfall,
RWE, Statoil, Dillinger Hu¨ttenwerke, Saarstahl and OMV. The pres-
ent study was undertaken in the frame of the R&D program GEO-
TECHNOLOGIEN (Publication No. GEOTECH-2096). Furthermore,
we would like to express our gratitude to the CO2MAN project part-
ners for data provision and scientific discussions. Discussion and conclusions Special thanks go to
Ben Norden (GFZ Potsdam) and Peter Frykman (GEUS Denmark) for
implementation and provision of the latest static geological 3D model,
to Benjamin Nakaten (GFZ Potsdam) for the technical implementation
of our Petrel-to-TOUGH2-MP workflow as well as to Elena Tillner
(GFZ Potsdam) for her support in model gridding. In addition, we
would also like to thank the four anonymous reviewers for their
constructive comments supporting the manuscript revision. better agreement with the measured data after each iter-
ation round. This workflow provides us with a static
model of advanced accuracy for further applications, for
prediction of long-term site behaviour (e.g. pressure
development and CO2 migration) as well as guiding
monitoring campaigns and site operation. Furthermore,
we were able to demonstrate the applicability of industrial
as well as scientific numerical simulators to successfully
describe multi-phase flow processes in the Stuttgart
Formation. One of our next simulation tasks will be the prediction
of the CO2 plume shape and thickness for the upcoming
3D seismic repeat in fall 2012, whereby different mod-
elling groups will be involved with their (in-house) sim-
ulation
codes. Further
activities
will
consider
the
enhancement of our Petrel-to-TOUGH2-MP workflow by
local grid refinement (LGR, as available in Petrel and
ECLIPSE) to allow for a realistic lateral well discretisa-
tion while maintaining the identical grid as applied in the
ECLIPSE simulations. Implementation of data from core
analysis from the CO2 Ktzi 203/2012 borehole (located in
about 30 m distance of the CO2 Ktzi 201/2007 well on
the connecting line to the CO2 Ktzi 202/2007 well)
drilled in fall 2012 will allow further improvement of the
static geological model in terms of facies-based hetero-
geneity development. Open Access
This article is distributed under the terms of the
Creative Commons Attribution License which permits any use, dis-
tribution, and reproduction in any medium, provided the original
author(s) and the source are credited. Fischer S, Liebscher A, De Lucia M, Hecht L (2013) Reactivity of
sandstone and siltstone samples from the Ketzin pilot CO2
storage site—laboratory experiments and reactive geochemical
modelling. Environ Earth Sci (this issue)
Grandy J (1997) Efficient computation of volume of hexahedral cells.
DOE Scientific and Technical Information, Lawrence Livermore References Fischer S, Liebscher A, De Lucia M, Hecht L (2013) Reactivity of
sandstone and siltstone samples from the Ketzin pilot CO2
storage site—laboratory experiments and reactive geochemical
modelling. Environ Earth Sci (this issue) Grandy J (1997) Efficient computation of volume of hexahedral cells. DOE Scientific and Technical Information, Lawrence Livermore 123 123 Environ Earth Sci National Laboratory. http://www.osti.gov/bridge/purl.cover.jsp? purl=/632793-4p2OLa/webviewable/632793.pdf. Accessed 27 June
2013 Mo¨ller F, Liebscher A, Martens S, Schmidt-Hattenberger C, Ku¨hn M
(2012) Yearly operational datasets of the CO2 storage pilot site
Ketzin, Germany. In: Scientific technical report: data 12/06 (pii
0.2312/GFZ.b103-12066, online only) Kempka T, Ku¨hn M, Class H, Frykman P, Kopp A, Nielsen CM,
Probst P (2010) Modeling of CO2 arrival time at Ketzin—part I. Int J Greenh Gas Control 4(6):1007–1015. doi:10.1016/j.ijggc. 2010.07.005 Norden B, Frykman P (2013) Geological modeling of the Stuttgart
Formation at Ketzin, Germany. Int J Greenh Gas Control (in
press) doi:10.1016/j.ijggc.2013.04.019 Pruess K (2005) ECO2N: a TOUGH2 fluid property module for
mixtures of water, NaCl, and CO2. In: Report LBNL-57952,
Lawrence Berkeley National Laboratory, Berkeley Kempka T, Klein E, De Lucia M, Tillner E, Ku¨hn M (2013a)
Assessment of long-term CO2 trapping mechanisms at the Ketzin
pilot site (Germany) by coupled numerical modeling. Energy
Procedia (in press). doi:10.1016/j.egypro.2013.06.460 Schlumberger (2009) Eclipse reservoir engineering software version
2009.2 Kempka T, Class H, Go¨rke UJ, Norden B, Kolditz O, Ku¨hn M, Walter
L, Wang W, Zehner B (2013b) A dynamic flow simulation code
intercomparison based on the revised static model of the Ketzin
pilot site. Energy Procedia (EGU GA 2013, in press) Schlumberger (2010) Petrel seismic-to-evaluation software version
2010.2.2 Wiese B, Bo¨hner J, Enachescu C, Wu¨rdemann H, Zimmermann G
(2010) Hydraulic characterisation of the Stuttgart formation at
the pilot test site for CO2 storage, Ketzin, Germany. Int J Greenh
Gas Control 4(6):960–971. doi:10.1016/j.ijggc.2010.06.013 Klein E, De Lucia M, Kempka T, Ku¨hn M (2013) Evaluation of long-
term mineral trapping at the Ketzin pilot site for CO2 storage: an
integrative approach using geochemical modelling and reservoir
simulation. Int J Greenh Gas Control (in press) doi:10.1016/j. ijggc.2013.05.014 j jgg
Wiese B, Zimmer M, Nowak M, Pilz P (2013) Above-zone well-
based hydraulic and geochemical monitoring of the CO2
reservoir in Ketzin, Germany. Environ Earth Sci (this issue) Kunz O, Klimeck R, Wagner W, Jaeschke M (2007) The GERG-2004
wide-range equation of state for natural gases and other mixtures
(GERG TM15 2007). In: Progress report VDI, vol. 6, no. References 557,
VDI Verlag, Du¨sseldorf Wu¨rdemann H, Mo¨ller F, Ku¨hn M, Heidug W, Christensen NP, Borm
G, Schilling FR, the CO2SINK Group (2010) CO2SINK—from
site characterisation and risk assessment to monitoring and
verification: one year of operational experience with the field
laboratory for CO2 storage at Ketzin, Germany. Int J Greenh Gas
Control 4(6):938–951. doi:10.1016/j.ijggc.2010.08.010 Lengler U, De Lucia M, Ku¨hn M (2010) The impact of heterogeneity
on the distribution of CO2: numerical simulation of CO2 storage
at Ketzin. Int J Greenh Gas Control 4(6):1016–1025. doi:10. 1016/j.ijggc.2010.07.004 Zhang K, Wu Y-S, Pruess K (2008) User’s guide for TOUGH2-MP—
a massively parallel version of the TOUGH2 code. In: Report
LBNL-315E, Earth Sciences Division, Lawrence Berkeley
National Laboratory, Berkeley Martens S, Kempka T, Liebscher A, Lu¨th S, Mo¨ller F, Myrttinen A,
Norden B, Schmidt-Hattenberger C, Zimmer M, Ku¨hn M (2012)
Europe’s longest-operating on-shore CO2 storage site at Ketzin,
Germany: a progress report after three years of injection. Environ Earth Sci 67:323–334. doi:10.1007/s12665-012-1672-5 Zimmer M, Erzinger J, Kujawa C, CO2SINK Group (2011) The gas
membrane sensor (GMS): a new method for gas measurements
in deep boreholes applied at the CO2SINK site. Int J Greenh Gas
Control 5(4):995–1001. doi:10.1016/j.ijggc.2010.11.007 Michael K, Golab A, Shulakova V, Ennis-King J, Allinson G, Sharma
S, Aiken T (2010) Geological storage of CO2 in saline
aquifers—a review of the experience from existing storage
operations. Int J Greenh Gas Control 4(4):659–667. doi:10.1016/
j.ijggc.2009 12 123 12
|
https://openalex.org/W4236895746
|
https://bmcinfectdis.biomedcentral.com/track/pdf/10.1186/s12879-021-05920-3
|
English
| null |
Initiation of Hepatitis C treatment in two rural Rwandan districts: A mobile clinic approach
|
Research Square (Research Square)
| 2,020
|
cc-by
| 8,251
|
Kamali et al. BMC Infectious Diseases (2021) 21:220
https://doi.org/10.1186/s12879-021-05920-3 Kamali et al. BMC Infectious Diseases (2021) 21:220
https://doi.org/10.1186/s12879-021-05920-3 Open Access Initiation of hepatitis C treatment in two
rural Rwandan districts: a mobile clinic
approach Innocent Kamali1*, Dale A. Barnhart1,2, Françoise Nyirahabihirwe1, Jean de la Paix Gakuru1, Mariam Uwase1,
Esdras Nizeyumuremyi1, Stephen Walker3, Christian Mazimpaka1, Jean de Dieu Gatete1, Jean Damascene Makuza4,5,
Janvier Serumondo4, Fredrick Kateera1 and Jean d’Amour Ndahimana1 © The Author(s). 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License,
which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give
appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if
changes were made. The images or other third party material in this article are included in the article's Creative Commons
licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons
licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain
permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the
data made available in this article, unless otherwise stated in a credit line to the data. Abstract Keywords: Hepatitis C, Mobile clinic, Rural health, Rwanda * Correspondence: inocek1@gmail.com * Correspondence: inocek1@gmail.com
1Partners In Health/Inshuti Mu Buzima, Rwinkwavu, Rwanda
Full list of author information is available at the end of the article Correspondence: inocek1@gmail.com
1Partners In Health/Inshuti Mu Buzima, Rwinkwavu, Rwanda
Full list of author information is available at the end of the article Abstract Background: To eliminate hepatitis C, Rwanda is conducting national mass screenings and providing to people
with chronic hepatitis C free access to Direct Acting Antivirals (DAAs). Until 2020, prescribers trained and authorized
to initiate DAA treatment were based at district hospitals, and access to DAAs remains expensive and
geographically difficult for rural patients. We implemented a mobile clinic to provide DAA treatment initiation at
primary-level health facilities among people with chronic hepatitis C identified through mass screening campaigns
in rural Kirehe and Kayonza districts. Methods: The mobile clinic team was composed of one clinician authorized to manage hepatitis, one lab
technician, and one driver. Eligible patients received same-day clinical consultations, counselling, laboratory tests
and DAA initiation. Using clinical databases, registers, and program records, we compared the number of patients
who initiated DAA treatment before and during the mobile clinic campaign. We assessed linkage to care during the
mobile clinical campaign and assessed predictors of linkage to care. We also estimated the cost per patient of
providing mobile services and the reduction in out-of-pocket costs associated with accessing DAA treatment
through the mobile clinic rather than the standard of care. Results: Prior to the mobile clinic, only 408 patients in Kirehe and Kayonza had been initiated on DAAs over a 25-
month period. Between November 2019 and January 2020, out of 661 eligible patients with hepatitis C, 429 (64.9%)
were linked to care through the mobile clinic. Having a telephone number and complete address recorded at
screening were strongly associated with linkage to care. The cost per patient of the mobile clinic program was
29.36 USD, excluding government-provided DAAs. Providing patients with same-day laboratory tests and clinical
consultation at primary-level health facilities reduced out-of-pocket expenses by 9.88 USD. Conclusion: The mobile clinic was a feasible strategy for providing rapid treatment initiation among people
chronically infected by hepatitis C, identified through a mass screening campaign. Compared to the standard of
care, mobile clinics reached more patients in a much shorter time. This low-cost strategy also reduced out-of-
pocket expenditures among patients. However, long-term, sustainable care would require decentralization to the
primary health-centre level. Introduction care for patients with hepatitis C in Rwanda’s Kirehe
and Kayonza districts. This paper describes the mobile
clinic model and implementation experience, coverage,
and costs associated with running the hepatitis C mobile
clinic. Chronic hepatitis C infection affects 71 million people
globally [1]. Untreated chronic hepatitis C can lead to
cirrhosis of the liver, liver failure, and hepatocellular car-
cinoma, making hepatitis C one of the leading causes of
liver cirrhosis and deaths [2]. Although Direct Acting
Antiretroviral (DAA) treatment has been shown to cure
over 90% of chronic hepatitis C cases [3–6] only 14 mil-
lion people infected with hepatitis C virus know their
status and only 1.1 million have initiated treatment [1]. In this context, the World Health Organization (WHO)
has developed a campaign to eliminate hepatitis C by
2030 [7]. © The Author(s). 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License,
which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give
appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if
changes were made. The images or other third party material in this article are included in the article's Creative Commons
licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons
licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain
permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the
data made available in this article, unless otherwise stated in a credit line to the data. Page 2 of 9 Kamali et al. BMC Infectious Diseases (2021) 21:220 Page 2 of 9 Methods
Setting g
Kayonza and Kirehe are located in Rwanda’s eastern
province, and are two of the three districts supported by
Partners In Health/Inshuti Mu Buzima (PIH/IMB), a
non-government organization that has been supporting
Rwandan Ministry of Health since 2005 in health system
strengthening. PIH/IMB supports Rwinkwavu and Kir-
ehe district hospitals together with their 25 affiliated
health centres, eight from Rwinkwavu and seventeen
from Kirehe. On average, health centres are about 24 km
from the hospital, which is over 8 h round trip on foot. Kayonza and Kirehe districts have hepatitis C antibody
(Ab) positivity rates of 7.7 and 11.6%, respectively [16]. Mass screening campaigns were conducted in Kirehe
and Kayonza in September 2019. Any patients testing
positive for hepatitis C antibodies using a rapid diagnos-
tic test and capillary blood were provided with a con-
firmatory Ribonucleic Acid (RNA) test using a venous
blood sample and all patients with a detectable viral load
(> 15 IU/L) were eligible for DAA treatment initiation. In sub-Saharan Africa, seroprevalence of hepatitis C is
estimated at around 3% [8], which is approximately three
times higher than estimates from general populations in
Europe (0.54–1.5%) [9] and the United States (US)
(0.9%) [10]. Rwanda, where hepatitis C prevalence esti-
mates range between 6.8 and 9% among people over 25
years old and above, [11, 12] is the first country in sub-
Saharan Africa to launch a national hepatitis C elimin-
ation plan. This ambitious plan exceeds WHO targets
[7] and aims to screen 4 million people and treat at least
90% of identified cases by 2024 [13]. In response to this
call to action, the Rwandan government has expanded
access to rapid diagnostic testing (RDT) through na-
tional mass screening campaigns and has guaranteed
free access to DAA medication for all people with
chronic hepatitis. However, ensuring adequate linkage to
care and treatment requires the decentralization of clini-
cians who are trained in hepatitis management. When
Rwanda introduced the first national hepatitis preven-
tion and management program in 2015, only 3 pharma-
cies in Rwanda were authorized to dispense drugs for
hepatitis, all located in Kigali city [14]. Data analysis To minimize turn-around time,
we
made
efforts
to
perform
biochemistry
and
hematology tests in a single batch each day for all
patients for a maximum of one hour and half be-
tween sample collection and result availability. Cli-
nicians
performed
medical
examinations
that
included
physical
consultations;
nutritional
status
assessment through anthropometric measurements;
evaluation of hepatitis C risk factors and patient
family history; and assessment of extrahepatic mani-
festations
such
as
skin
rash,
vasculitis,
and
co-
morbidities. Clinicians used the liver function and
hematology test results to calculate an Aspartate
Aminotransferase
to
Platelet
Ratio
Index
(APRI)
score [17]. Patients with an APRI< 2 and no other
clinical signs of decompensation received immediate
treatment initiation. Patients with APRI score > 2 or
other clinical signs of decompensation were referred
to a specialist for liver ultrasound and hepatocellu-
lar carcinoma screening, per the national guidelines. Pregnant and breastfeeding women deferred treat-
ment initiation until after they completed breast-
feeding. Socio-demographic and clinical information
were recorded in a paper-based patient file printed
by the national hepatitis program. Overall, patients
spent four to five hours at the mobile clinic to ac-
cess all services. To facilitate patient follow-up, data
from the hepatitis mass screening campaign and
paper-based patient files were entered into a dedi-
cated REDCap database used by PIH/IMB to man-
age hepatitis patients [18]. This study was approved
by
the
Inshuti
Mu
Buzima
Research
Committee
(IMBRC) and Rwanda National Ethics Committee
(RNEC) 015/RNEC/2020) and all methods were per-
formed in accordance with local guidelines and reg-
ulations. Because this study used retrospective data
which was collected as part of routine clinical prac-
tice, informed consent was not obtained. We used demographic data collected during the
mass screening campaigns to compare characteristics
of eligible patients who were and who were not able
to be linked to care through the mobile clinic cam-
paign using a Pearson’s chi-squared test. We reported
risk ratios and 95% confidence intervals comparing
the probability of being reached by the mobile clinic
among patients with and without telephone number
information and among patients with and without
complete information on their address, defined as
having information listed on the district, sector, cell,
and village. This contact information was assessed for
completeness, not for accuracy. All statistical analyses
were conducted using Stata v.15.1 (Stata Corp, Col-
lege Station, TX, USA). Data analysis driver. We also provided two mobile laboratory ma-
chines, a Humalyser 3500 for biochemistry and Sysmex
XP300 for hematology. We provided all necessary re-
agents to complete the pre-treatment initiation tests on
the mobile machines, including Alanine Aminotransami-
nase (ALT), Aspartate Aminotransferase (AST), cell
pack, and Stromatolyzer-WH; as well as other necessary
medical supplies and commodities. Although the clinical
team and laboratory machines were mobile, all clinical
activities took place in the existing health centre infra-
structure. All lab tests performed at the mobile clinic
used venous blood samples. driver. We also provided two mobile laboratory ma-
chines, a Humalyser 3500 for biochemistry and Sysmex
XP300 for hematology. We provided all necessary re-
agents to complete the pre-treatment initiation tests on
the mobile machines, including Alanine Aminotransami-
nase (ALT), Aspartate Aminotransferase (AST), cell
pack, and Stromatolyzer-WH; as well as other necessary
medical supplies and commodities. Although the clinical
team and laboratory machines were mobile, all clinical
activities took place in the existing health centre infra-
structure. All lab tests performed at the mobile clinic
used venous blood samples. y
We identified the number of people with chronic hepa-
titis C who had initiated on DAAs prior to the mobile
clinic using data extracted from patient registries located
at Kirehe and Rwinkwavu district hospitals. We assumed
any patient who initiated hepatitis C treatment in Kirehe
between November 18th, 2019 and January 31st, 2020 or
in Kayonza between December 12th, 2019 and January
31st 2020, was a beneficiary of the mobile clinic cam-
paign. To estimate coverage of the mobile clinic cam-
paign, we identified patients who were eligible for
treatment initiation through the mobile clinic using data
extracted from the REDCap database. We defined pa-
tients as eligible for treatment initiation through the mo-
bile model if they were: a) living in the catchment area
of a PIH/IMB-supported health facility in Kayonza or
Kirehe district, b) either screened positive for hepatitis C
before January 31st, 2020 or had viral load test results
indicating a detectable viral load for hepatitis C dated
prior to January 31st, 2020, and c) had not started treat-
ment prior to the start of the mobile clinic campaign in
each district. At the mobile clinic, we assessed patient attend-
ance and the clinician provided pre-treatment coun-
selling in a group setting. Next, patients were sent
to the laboratory technician for liver function and
hematology tests. Description of the intervention p
We developed the mobile hepatitis clinic to provide pa-
tients with access to DAAs treatment at primary-level
health facility. Using a list of patients identified as eli-
gible for hepatitis C treatment during the previous mass
screening campaigns, we made a schedule to visit each
health centre within the target districts. Patients who
had screened positive for hepatitis B were also linked to
care during the mobile clinic, but these patients reflected
a minority of the patients linked to care were not in-
cluded in this analysis unless they were co-infected with
hepatitis C. Prior to the day of the mobile clinic visit,
health centre staff contacted patients via telephone to
schedule appointments. When patients could not be
reached via telephone, we used the existing network of
village-level community health workers to reach patients
through home visits and inform the patient of the mo-
bile clinic date. We also contacted each district phar-
macy to ensure that a twelve-week course of DAA
treatment could be reserved for all eligible patients to
avoid any possible stock-outs. Clinicians were able to
prescribe and dispense DAAs directly to the patients. On the mobile clinic day, we deployed a multidisciplin-
ary team composed of (i) one physician or a nurse au-
thorized
by
the
Ministry
of
Health
for
hepatitis
management, (ii) one laboratory technician, and (iii) a By June 2018 the Rwandan government had trained in-
fectious diseases clinical mentors, which included both
medical doctors and nurses from district hospitals, and
lab technicians to provide hepatitis management services
at all district hospitals around the country [15]. Per na-
tional guidelines, both doctors and nurses are able to
manage hepatitis B and C including clinical consulta-
tions, request for lab exams, treatment prescription, drug
distribution and follow-up. However, accessing hepatitis
C treatment can still be difficult and expensive for rural
Rwandans. In addition to expenses related to traveling to
and from the district hospitals, patients must also pay
out-of-pocket for liver function and hematology labora-
tory tests, which are needed before DAAs initiation. Typically, laboratory tests and treatment initiation con-
sultations occur on different days, increasing the trans-
portation costs for patients. In order to reduce expenses
and travel-related barriers to treatment initiation, we de-
veloped a mobile hepatitis clinic to improve access to Kamali et al. BMC Infectious Diseases (2021) 21:220 Kamali et al. BMC Infectious Diseases (2021) 21:220 Page 3 of 9 Cost of running the mobile clinics Cost of running the mobile clinics
As shown in Table 2, the total cost per patient initiated
on hepatitis C treatment, including the cost of DAAs,
was $89.36. DAAs reflected 67% of the total program
cost while mobile clinic outreach activities cost $29.36
per person, reflecting only 32.8% of the total cost of the
program. Besides DAAs, the largest costs associated with
mobile
clinic
implementation
were
overhead
and
transportation. Data analysis All costs were converted from Rwandan francs
to U.S. dollars using an exchange rate of 920 francs per
dollar. To estimate the reduction in patients’ out-of-
pocket expenses associated with initiating treatment
through the mobile clinic rather than through the stand-
ard of care, we assumed that the major differences in
out-of-pocket expenses would be the cost of transport
from the local health centre to a district hospital and the
cost of covering pre-treatment laboratory tests, which
are not currently covered by Rwanda’s Community
Based Health Insurance program (CBHI/Mutuelle). We
used the cost of lab tests under CBHI/Mutuelle in our
primary analysis because it is the most affordable and
widely subscribed to insurance program and covers
81.6% of Rwandans [21]. However, we also report the es-
timated reduction in out-of-pocket costs among the un-
insured. We did not include the cost of transport
from the home to the local health centre because
most patients travel to health centres on foot and be-
cause this cost would be the same both before and
after the initiation of the mobile clinic. We estimated
an average round trip cost of transportation from
health facilities to district hospitals in Kayonza and
Kirehe districts using standard PIH/IMB transporta-
tion reimbursement levels. We used the current prices
of medical services offered at district hospitals to esti-
mate the out-of-pocket costs for necessary laboratory
tests under CBHI/Mutuelle and among the uninsured Results participate in the campaign by the number of days of the
campaign. To estimate the cost of the driver, we allo-
cated the driver’s gross annual salary proportionally to
the number of days dedicated to the mobile clinic cam-
paign. The cost of capital, including vehicles and labora-
tory
machines,
was
estimated
by
calculating
the
annualized purchase price of these items over the antici-
pated lifetime of the item assuming an annual discount
rate of 10%. To calculate the daily cost of these items,
we divided the annual cost by 260 to reflect the number
of working days in a year. The cost of supplies and fees
were
identified
from
the
mobile
clinic’s
operating
budget. We estimated the cost of DAA medication based
on the government of Rwanda’s negotiated price with
the drug manufacturer [20]. We reported the cost-per-
patient of our program both with and without the cost
of DAAs. In Rwanda, these are provided by the govern-
ment free of charge, and providing mobile clinic services
to patients does not include the price of these drugs. However, we recognize that in other settings the mobile
clinic team may also be responsible for covering the cost
of drugs. After identifying the total cost of running the
mobile clinic, we divided the cost by the total number of
patients with hepatitis C initiated to estimate cost per
patient. All costs were converted from Rwandan francs
to U.S. dollars using an exchange rate of 920 francs per
dollar. To estimate the reduction in patients’ out-of-
pocket expenses associated with initiating treatment
through the mobile clinic rather than through the stand-
ard of care, we assumed that the major differences in
out-of-pocket expenses would be the cost of transport
from the local health centre to a district hospital and the
cost of covering pre-treatment laboratory tests, which
are not currently covered by Rwanda’s Community
Based Health Insurance program (CBHI/Mutuelle). We
used the cost of lab tests under CBHI/Mutuelle in our
primary analysis because it is the most affordable and
widely subscribed to insurance program and covers
81.6% of Rwandans [21]. However, we also report the es-
timated reduction in out-of-pocket costs among the un-
insured. Results We did not include the cost of transport
from the home to the local health centre because
most patients travel to health centres on foot and be-
cause this cost would be the same both before and
after the initiation of the mobile clinic. We estimated
an average round trip cost of transportation from
health facilities to district hospitals in Kayonza and
Kirehe districts using standard PIH/IMB transporta-
tion reimbursement levels. We used the current prices
of medical services offered at district hospitals to esti-
mate the out-of-pocket costs for necessary laboratory
tests under CBHI/Mutuelle and among the uninsured
[22]. Costing analyses were conducted using Microsoft Data analysis To estimate the cost per patient of delivering mobile
clinic services, we used an ingredients-based approach
and a healthcare provider perspective. Following recom-
mendations from the WHO Guide to Cost-Effectiveness
Analysis [19], we included the costs of items typically
covered by the PIH/ IMB operating budget in our ana-
lysis to reflect the opportunity cost of using these re-
sources for the hepatitis C mobile clinic rather than for
other activities. We categorized costs as either overhead,
mobile clinic staff, capital, supplies and fees, and medica-
tion. To estimate overhead costs, we identified PIH/IMB
permanent staff who were involved in organizing the
campaign and allocated the value of their gross annual
salary proportionally to the number of days they dedi-
cated to the campaign. To reflect the cost of mobile
clinic staff, we multiplied the daily salaries for the tem-
porary laboratory technician and clinicians contracted to Kamali et al. BMC Infectious Diseases (2021) 21:220 Kamali et al. BMC Infectious Diseases (2021) 21:220 Page 4 of 9 Page 4 of 9 participate in the campaign by the number of days of the
campaign. To estimate the cost of the driver, we allo-
cated the driver’s gross annual salary proportionally to
the number of days dedicated to the mobile clinic cam-
paign. The cost of capital, including vehicles and labora-
tory
machines,
was
estimated
by
calculating
the
annualized purchase price of these items over the antici-
pated lifetime of the item assuming an annual discount
rate of 10%. To calculate the daily cost of these items,
we divided the annual cost by 260 to reflect the number
of working days in a year. The cost of supplies and fees
were
identified
from
the
mobile
clinic’s
operating
budget. We estimated the cost of DAA medication based
on the government of Rwanda’s negotiated price with
the drug manufacturer [20]. We reported the cost-per-
patient of our program both with and without the cost
of DAAs. In Rwanda, these are provided by the govern-
ment free of charge, and providing mobile clinic services
to patients does not include the price of these drugs. However, we recognize that in other settings the mobile
clinic team may also be responsible for covering the cost
of drugs. After identifying the total cost of running the
mobile clinic, we divided the cost by the total number of
patients with hepatitis C initiated to estimate cost per
patient. Number of patients initiated before and during the
mobile clinic Number of patients initiated before and during the
mobile clinic Between the availability of hepatitis C treatment pro-
gram in Kirehe and Rwinkwavu district hospitals in Sep-
tember 2017 and the start of the mobile clinic program,
408 patients initiated treatment for hepatitis C. Two pa-
tients from Kayonza and 13 from Kirehe exhibited APRI
score > 2 or other clinical signs of decompensation were
referred to a specialist for liver ultrasound and hepato-
cellular carcinoma screening, per the national guidelines. Overall, the 429 of patients initiated during eleven-week
period of mobile clinic campaign exceeded the total
number of patients initiated during the previous 25
months of the hepatitis treatment program (Fig. 1). Coverage of mobile clinic campaign Using the REDCap database, we identified 661 patients
with chronic hepatitis C in Southern Kayonza and Kir-
ehe districts who were eligible for hepatitis C treatment
initiation during the mobile clinic campaign. The 429
patients who were linked to care reflect program cover-
age of 64.9% (95% CI: 61.1–68.5%). Mobile clinic cover-
age was 71.3% in Southern Kayonza and 62.4% in Kirehe
(Table 1). There was no difference in coverage between
males and females, age, socioeconomic status, insurance
status, marital status, or profession. Patients with their full address recorded during screen-
ing were 3.10 times more likely to be reached during the
mobile clinic (95% CI: 2.32, 4.16) while patients who had
a telephone number recorded at screening were 1.79
times more likely to be reached during the mobile clinic
(95% CI: 1.51–2.11). In general, missing data was much
less common among the 416 patients identified during
the September 2019 mass screening campaigns than
among the 245 patients identified in previous campaigns. Data from these previous campaigns exhibited 61.2%
missingness for telephone number, 40.8% missingness
for complete address, and over 50% missingness for ubu-
dehe, insurance status, marital status, and profession. Out of pocket costs for patients When estimating the expected out-of-pocket expenses
among patients seeking to initiate DAA treatment, we Kamali et al. BMC Infectious Diseases (2021) 21:220 Page 5 of 9 Fig. 1 Comparison of patients with hepatitis C initiated before and during mobile clinic elsewhere in Rwanda and to other countries seeking to
scale up access to hepatitis C treatment. found that patients covered using CBHI/Mutuelle pro-
gram could expect to pay $ 9.87 more in out-of-pocket
under the standard of care compared to under the mo-
bile clinic model. For patients without CBHI/Mutuelle,
the reduction in out-of-pocket expense was estimated at
$19.71 (Table 3). This reduction is an underestimation
of the true cost savings for the patients as it does not in-
clude the reduction in opportunity costs associated with
decreasing the number of days patients spent in the
clinic from two to one. When including the cost of the DAAs in our program,
we found that the costs of implementing the mobile
clinic system reflected only 32.8% of the total cost per
patient initiated. In Rwanda, where the national govern-
ment is committed to providing DAAs free-of-charge to
all patients with hepatitis C, this relatively small increase
in per-patient costs may be an important investment to
ensure adequate linkage to care and equitable access to
treatment for all citizens. Although we did not conduct
a formal cost-effectiveness analysis, the per-patient cost
of this program compares favorably with an antenatal
care program in Rwanda where the first visit costs $21
per woman [23]. Discussion Our
experience
demonstrates
that
mobile
hepatitis
clinics are a feasible treatment strategy to promote
same-day treatment initiation for patients with hepatitis
C in resource-constrained settings. Through our mobile
clinics, we were able to initiate 64.9% of all patients
awaiting treatment on DAAs during an eleven- week
period. The number of patients initiated on treatment
through the mobile clinic program exceeded the total
number of patients who were initiated on treatment
under the standard of care during the previous 25
months. While some of this difference may reflect ex-
panded testing capacity in Rwanda as new initiatives that
have been introduced, for example the adoption of rapid
diagnostic tests and same-day venous blood collection
for viral load testing during the mass screening cam-
paigns, our results demonstrate that mobile clinics can
be used to ensure prompt linkage to care where services
are not decentralized. This strategy is especially useful
following mass screening campaigns, when large num-
bers of patients are awaiting treatment initiation. To our
knowledge, this is the first hepatitis C treatment mobile
clinic program model implemented in Rwanda and sub-
Saharan Africa. Our program may serve as a model The cost of our program also compares favorably to
the cost of HIV care and treatment visits and antiretro-
viral therapy, which is estimated to cost an average of
$208 per patient per year in Rwanda, Ethiopia, Malawi
and Zambia [24]. Mobile clinics have successfully increased access to
care in rural settings for other programs, including pre-
natal care, HIV, and other sexually transmitted infec-
tions [25]. Mobile clinics can be used to both reduce
patient costs and improve health outcomes in under-
served and vulnerable populations [26]. Since 83% of
Rwandans live in rural areas [27] many of them have to
walk long distances or arrange costly transport to access
health services at district hospitals. We estimated that
our mobile clinic program was able to cut transport time
in half and reduce patients’ out-of-pocket expenses by
$9.87. This is a meaningful cost reduction in a country
where 43.1% of the rural population live under poverty
and 18.1% in an extreme poverty [22]. Over one third of
the reduction in out-of-pocket costs is attributable to Kamali et al. BMC Infectious Diseases (2021) 21:220 Page 6 of 9 our provision of free liver and renal function tests
through the mobile clinic program. Discussion All costs given in US dollars
Total
Administration and Overhead1
5606.45
Mobile Clinic Staff
Daily Salary
Days
Clinician
15.53
34
527.92
Lab Tech
15.53
34
527.92
Driver
13.53
34
460.06
Capital
Annualized Daily Cost
Number of Units
Vehicles2
38.48
34
1308.32
Human Biochemistry 35003
2.04
34
69.40
Sysmex XP 300 Hemotology4
6.12
34
208.19
Fees, Supplies, and Materials
Cost per Unit
Number of Units
Per diem fees for PIH staff
2.18
102
222.36
Per diem fees for HC staff
15.53
68
1056.04
Daily fuel
11.96
34
406.64
Reagents
ALAT
1.19
429
510.51
ASAT
1.19
429
510.51
Hematology
2.11
429
905.10
Other commodities
Alcohol Swabs
0.33
429
139.89
Cotton
0.11
429
46.63
Gloves
0.13
429
55.96
Needles
0.07
429
28.91
Tubes
0.01
429
4.29
Medication
Cost per Unit
Number of Units
DAAS
60.00
429
25,740.00
TOTAL COST OF PROGRAM
38,335.11
TOTAL COST PER PATIENT
89.36
TOTAL COST PER PATIENT, EXCLUDING DAAS
29.36
1Overhead was calculated by allocating annual gross of salaries of ID team members proportionally to the total number of days each permanent ID staff member
dedicated to this project
2Annualized cost of a vehicle assumes a purchase price of 76,087 an annual discount rate of 10 and a useful lifespan of 15 years
3 Annualized cost of Human Biochemistry 3500 machine assumes a purchase price of 3260 an annual discount rate of 10% and a useful lifespan of 10 years
4Annualized cost of Sysmex XP 300 Hemotology machine assumes a purchase price of 9782 an annual discount rate of 10% and a useful lifespan of 10 years 1Overhead was calculated by allocating annual gross of salaries of ID team members proportionally to the total number of days each permanent ID staff membe
dedicated to this project
2Annualized cost of a vehicle assumes a purchase price of 76,087 an annual discount rate of 10 and a useful lifespan of 15 years
3 Annualized cost of Human Biochemistry 3500 machine assumes a purchase price of 3260 an annual discount rate of 10% and a useful lifespan of 10 years
4Annualized cost of Sysmex XP 300 Hemotology machine assumes a purchase price of 9782 an annual discount rate of 10% and a useful lifespan of 10 years linked to care during the mobile clinic campaign. Discussion To strengthen sup-
Mutuelle package at health centre for patients with viral
hepatitis. Table 1 Characteristics of patients. The overall samples size was 661. Sample sizes are given for each variable to reflect missing data
Factor
Not reached by mobile clinic
Reached by mobile clinic
p-value
N = 232
N = 429
PIH Supported Site
0.030
Kayonza
54 (28.7%)
134 (71.3%)
Kirehe
178 (37.6%)
295 (62.4%)
Sex (N = 655)
0.77
Female
160 (35.5%)
291 (64.5%)
Male
70 (34.3%)
134 (65.7%)
Age (years), median IQR
60 (48–72)
62 (50.5–70)
0.46
Ubudehe (N = 474)
0.16
Category 1
25 (26.3%)
70 (73.7%)
Category 2
29 (17.2%)
140 (82.8%)
Category 3
50 (24.4%)
155 (75.6%)
Not known
0 (0.0%)
5 (100.0%)
Insurance Status (N = 501)
0.54
No insurance
0 (0.0%)
1 (100.0%)
CBHI/Mutuelle
74 (15.1%)
416 (84.9%)
RSSB
0 (0.0%)
9 (100.0%)
Other
0 (0.0%)
1 (100.0%)
Marital Status (N = 492)
0.74
Single
8 (21.1%)
30 (78.9%)
Married or Cohabitating
40 (14.6%)
234 (85.4%)
Widowed
25 (14.6%)
146 (85.4%)
Divorced
1 (11.1%)
8 (88.9%)
Profession category (N = 449)
0.22
Farmer
59 (14.3%)
353 (85.7%)
Unskilled Labor
2 (13.3%)
13 (86.7%)
Skilled Labor
0 (0.0%)
2 (100.0%)
Professional
0 (0.0%)
12 (100.0%)
Student
3 (37.5%)
5 (62.5%)
Telephone recorded at screening (N = 661)
< 0.001
No
123 (57.7%)
90 (42.3%)
Yes
109 (24.3%)
339 (75.7%)
Full address collected at screening (N = 661)
< 0.001
No
108 (75.5%)
35 (24.5%)
Yes
124 (23.9%)
394 (76.1%)
Screened during September 2019
< 0.001
Yes
59 (14.2%)
357 (85.8%)
No
173 (70.6%)
72 (29.4%) Mutuelle package at health centre for patients with viral
hepatitis. Mutuelle package at health centre for patients with viral
hepatitis. our provision of free liver and renal function tests
through the mobile clinic program. To strengthen sup-
port for vulnerable patients and to promote the goals of
the national hepatitis C elimination campaign, the gov-
ernment of Rwanda may consider including liver and
renal function tests as a covered service in CBHI/ During the implementation of the mobile clinic, not
all expected patients could be reached to schedule their
appointment. Although we worked with community
healthcare workers to seek patients in their home Kamali et al. BMC Infectious Diseases (2021) 21:220 Page 7 of 9 Table 2 Cost of hepatitis C mobile clinic from the health care provider perspective. Discussion All costs given in US dollars ost of hepatitis C mobile clinic from the health care provider perspective. All costs given in US dollars Table 2 Cost of hepatitis C mobile clinic from the health care provider perspective. Discussion To im-
prove the effectiveness of these mass-screening cam-
paigns, we recommend the adoption of high-quality
training for data collectors, investment in an electronic
data capturing system, and real-time monitoring and villages, some patients were impossible to contact, pos-
sibly because information was entered incorrectly during
the mass screening campaigns. As evidenced by our ana-
lysis, having complete data on telephone number and
address were strongly associated with being able to be Table 3 Reduction in out of pocket expenses for mobile clinic patients (USD)
Item
Cost reduction with CBHI/Mutuelle
Cost reduction without CBHI/Mutuelle
Transport from health centre to hospital
$5.86
$5.86
ALT
$1.19
$4.11
AST
$1.19
$4.11
Hematology test
$1.63
$5.62
Total
$9.87
$19.71 Table 3 Reduction in out of pocket expenses for mobile clinic patients (USD) Kamali et al. BMC Infectious Diseases (2021) 21:220 Page 8 of 9 Kamali et al. BMC Infectious Diseases (2021) 21:220 Page 8 of 9 did not include explicit information about mobile clinic
participation, we had to rely on dates of screening, viral
load testing, and treatment initiation to assess mobility
clinic eligibility and participation. However, because we
were the only health care provider offering hepatitis
treatment in our catchment area and because our results
are very similar to daily records kept by mobile clinic
staff during the campaign, we believe any misclassifica-
tion is minimal. As part of our campaign, we also initi-
ated some patients who had screened positive for
hepatitis B; however, we did not include these patients
in this analysis. This decision reflects the fact that, unlike
hepatitis C, hepatitis B does not currently have a cure,
requires life-long treatment, and is less suited to short-
term campaigns than hepatitis C. Including patients with
hepatitis B in our costing analysis would have reduced
the per-patient costs of the overall campaign. Finally, we
did not compare the quality of care provided during mo-
bile clinics to what was provided prior to the initiation
of the mobile clinic program. Despite these limitations,
we believe this analysis demonstrates the feasibility of a
mobile clinic-based model for hepatitis C treatment ini-
tiation, and hope that it can be used to inform future in-
terventions in similar contexts. feedback to ensure that data collectors are collecting in-
formation as accurately as possible. Discussion When we have been
able to implement these strategies in subsequent cam-
paigns, we have found that it has substantially improved
our ability to link patients to care, as partially evidenced
by the lower levels of missing data during the September
2019 campaign compared to previous campaigns. Im-
portantly, we did not observe differences in mobile clinic
coverage by age, gender, or socioeconomic status, sug-
gesting coverage of the mobile clinic program was rela-
tively equitable across demographic groups. We
identified
several
additional
lessons
learned
while implementing the mobile clinic campaign. First,
we were unable to provide ultrasound exams for pa-
tients with hepatitis who presented with suspected
liver decompensation. Although there were very few
cases with suspected liver decompensation, the avail-
ability of a mobile ultrasound machine could have im-
proved our ability to provide same-day initiation for
these patients. Second, clearly communicating with
the patients the starting hour for the mobile clinic
was
critical
for
enabling
collective
pre-treatment
counselling, processing biochemistry and hematology
tests in one batch, and ensuring that teams had
enough time to initiate all patient by the end of the
day. Finally, our mobile clinics were implemented fol-
lowing a mass screening campaign that identified a
large number of patients who required treatment ini-
tiation in a short period of time. However, the cost-
effectiveness of this strategy depends on having a
relatively large number of patients awaiting treatment
per health centre, as would typically be the case after
a mass screening campaign. Ultimately ensuring long-
term,
sustainable,
decentralized
access
to
hepatitis
treatment requires task shifting, where health centre-
level nurses are trained to manage hepatitis on a daily
basis. Decentralization of care has been demonstrated
model to be feasible by the HIV task-shifting model,
where nurses who have been well trained, mentored,
and given support can effectively manage HIV treat-
ment [28, 29]. Abbreviations
ALT Al
i
t ALT: Alanine transaminase; AST: Aspartate transaminase; APRI: Aspartate
Aminotransferase to Platelet Ratio Index; CBHI: Community Health Insurance;
CI: Confidence Interval; DAAs: Direct Acting Antivirals; IHDPC: Institute of HIV
Disease Prevention and Control; IMBRC: Inshuti Mu Buzima Research
Committee; IQR: Interquartile range; PIH/IMB: Partners In Health/Inshuti Mu
Buzima; MoH: Ministry of Health; RDT: Rapid Diagnostic Test; RBC: Rwanda
Biomedical Centre; REDCap: Research Electronic Data Capture;
RNA: Ribonucleic Acid; RNEC: Rwanda National Ethics Committee; USD: US
dollar; WHO: World Health Organization During our campaign, we did provide infectious dis-
ease nurses working at health centres with training on
hepatitis management, and, in collaboration with the
Ministry of Health, we have also been supporting theor-
etical and practical training sessions to allow these
nurses to become certified in hepatitis management. Our analysis has some limitations. We used clinical re-
cords to assess the number of patients initiated to treat-
ment before and during the period of mobile clinic
geographic and budgetary data to assess costs associated
with the program. These data sources were not intended
for research purposes and may suffer from missing or
incomplete data. Furthermore, because medical records Conclusion Access to hepatitis C treatment in Rwanda is improving
but is still limited in rural settings. Implementing a mo-
bile clinic program with basic laboratory services is a
feasible and potentially scalable tool to increase access
to treatment. This low-cost strategy can complement to
mass screening campaigns by linking large numbers of
patients to care in a short period of time. The model
also reduces time spent at a health facility and out-of-
pocket expenditures for patients. The model could po-
tentially be extended to other settings or to other dis-
eases requiring linkage to care for short-term curable
diseases in rural settings. ALT: Alanine transaminase; AST: Aspartate transaminase; APRI: Aspartate
Aminotransferase to Platelet Ratio Index; CBHI: Community Health Insurance;
CI: Confidence Interval; DAAs: Direct Acting Antivirals; IHDPC: Institute of HIV
Disease Prevention and Control; IMBRC: Inshuti Mu Buzima Research
Committee; IQR: Interquartile range; PIH/IMB: Partners In Health/Inshuti Mu
Buzima; MoH: Ministry of Health; RDT: Rapid Diagnostic Test; RBC: Rwanda
Biomedical Centre; REDCap: Research Electronic Data Capture;
RNA: Ribonucleic Acid; RNEC: Rwanda National Ethics Committee; USD: US
dollar; WHO: World Health Organization References Effectiveness of direct-acting antiviral therapy for hepatitis C in difficult-to-
treat patients in a safety-net health system: a retrospective cohort study. BMC Med. 2017;15(1):204. 29. Shumbusho F, van Griensven J, Lowrance D, Turate I, Weaver MA, Price J,
et al. Task Shifting for Scale-up of HIV Care: Evaluation of Nurse-centered
Antiretroviral Treatment at Rural Health Centres in Rwanda. Feeley F,. PLoS
Med. 2009;6(10):e1000163. 29. Shumbusho F, van Griensven J, Lowrance D, Turate I, Weaver MA, Price J,
et al. Task Shifting for Scale-up of HIV Care: Evaluation of Nurse-centered
Antiretroviral Treatment at Rural Health Centres in Rwanda. Feeley F,. PLoS
Med. 2009;6(10):e1000163. 6. Nsanzimana S, Penkunas MJ, Liu CY, Sebuhoro D, Ngwije A, Remera E, et al. Effectiveness of Direct-acting Antivirals for the Treatment of Chronic
Hepatitis C in Rwanda: A Retrospective Study. Clin Infect Dis. 2020;6:ciaa701. 7. Popping S, Bade D, Boucher C, van der Valk M, El-Sayed M, Sigurour O, et al. The global campaign to eliminate HBV and HCV infection: international viral
hepatitis elimination meeting and core indicators for development towards
the 2030 elimination goals. J Virus Erad. 2019;5(1):60–6. References 1. World Health Organization, World Health Organization, Global Hepatitis
Programme. Global hepatitis report, 2017 [Internet]. 2017 [cited 2020 Oct 6]. Available from: http://apps.who.int/iris/bitstream/10665/255016/1/9789241
565455-eng.pdf?ua=1 1. World Health Organization, World Health Organization, Global Hepatitis
Programme. Global hepatitis report, 2017 [Internet]. 2017 [cited 2020 Oct 6]. Available from: http://apps.who.int/iris/bitstream/10665/255016/1/9789241
565455-eng.pdf?ua=1 25. Kojima N, Krupp K, Ravi K, Gowda S, Jaykrishna P, Leonardson-Placek C, et al. Implementing and sustaining a mobile medical clinic for prenatal care and sexually
transmitted infection prevention in rural Mysore. India BMC Infect Dis. 2017;17(1):189. 2. Mokdad AA, Lopez AD, Shahraz S, Lozano R, Mokdad AH, Stanaway J, et al. Liver cirrhosis mortality in 187 countries between 1980 and 2010: a
systematic analysis. BMC Med. 2014;12(1):145. 2. Mokdad AA, Lopez AD, Shahraz S, Lozano R, Mokdad AH, Stanaway J, et al. Liver cirrhosis mortality in 187 countries between 1980 and 2010: a
systematic analysis. BMC Med. 2014;12(1):145. 26. Yu SWY, Hill C, Ricks ML, Bennet J, Oriol NE. The scope and impact of
mobile health clinics in the United States: a literature review. Int J Equity
Health. 2017;16(1):178. 26. Yu SWY, Hill C, Ricks ML, Bennet J, Oriol NE. The scope and impact of
mobile health clinics in the United States: a literature review. Int J Equity
Health. 2017;16(1):178. 3. Abozeid M, Alsebaey A, Abdelsameea E, Othman W, Elhelbawy M, Rgab A,
et al. High efficacy of generic and brand direct acting antivirals in treatment
of chronic hepatitis C. Int J Infect Dis. 2018;75:109–14. 27. National Institute of Statistics of Rwanda (NISR). Ministry of Finance and economic
planning. Rwanda: Forth Population and Housing Census; 2012. p. 2014. 27. National Institute of Statistics of Rwanda (NISR). Ministry of Finance and economic
planning. Rwanda: Forth Population and Housing Census; 2012. p. 2014. 4. Yang Y, Wu F-P, Wang W-J, Shi J-J, Li Y-P, Zhang X, et al. Real life efficacy
and safety of direct-acting antiviral therapy for treatment of patients
infected with hepatitis C virus genotypes 1, 2 and 3 in Northwest China. World J Gastroenterol. 2019;25(44):6551–60. 28. Nsanzimana S, Kirk C, Bucher H, Uwizihiwe J. Increasing Access to Hepatitis
C Treatment in Rwanda: The Promise of Rwanda’s Existing HIV Infrastructure. J Infect Dis Ther [Internet]. 2015 [cited 2019 Oct 10];03(05). Available from:
http://www.esciencecentral.org/journals/increasing-access-to-hepatitis-c-trea
tment-in-rwanda-the-promise-ofrwandas-existing-hiv-infrastructure-2090-
7214-1000245.php?aid=61773 5. Yek C, de la Flor C, Marshall J, Zoellner C, Thompson G, Quirk L, et al. Received: 9 December 2020 Accepted: 17 February 2021 Received: 9 December 2020 Accepted: 17 February 2021 24. Tagar E, Sundaram M, Condliffe K, Matatiyo B, Chimbwandira F, Chilima B,
et al. Multi-Country Analysis of Treatment Costs for HIV/AIDS: Facility-Level
ART Unit Cost Analysis in Ethiopia, Malawi, Rwanda, South Africa and
Zambia. Gantt S, PLoS ONE. 2014 ;9(11):e108304. Acknowledgements This mobile clinic would not have been implemented without the
contribution of healthcare providers from health facilities of Kirehe and
Rwinkwavu district hospitals and their affiliated health centres. We would like
to thank them. We also bring our thanks to the Ministry of Health and
Rwanda Biomedical Centre (RBC) for their special contribution in providing
drugs free of charge. Page 9 of 9 Page 9 of 9 Kamali et al. BMC Infectious Diseases (2021) 21:220 Kamali et al. BMC Infectious Diseases (2021) 21:220 Availability of data and materials
h d
f
h The datasets for this paper are not publicly available but are available from
the corresponding author on reasonable request. 14. Mbituyumuremyi A, Van Nuil JI, Umuhire J, Mugabo J, Mwumvaneza M,
Makuza JD, et al. Controlling hepatitis C in Rwanda: a framework for a
national response. Bull World Health Organ. 2018;96(1):51–8. national response. Bull World Health Organ. 2018;96(1):51–8. 15. Rwanda Biomedical Centre, Ministry of Health. Rwanda National HIV and Viral
Hepatitis Annual Report 2017-2018. Kigali: Rwanda Biomedical Centre; 2018. 16. Makuza JD. Training of health care providers on viral hepatitis prevention
d
M
R
d Bi
di
l C
2019 Funding
h
b 12. Umutesi J, Liu CY, Penkunas MJ, Makuza JD, Ntihabose CK, Umuraza S, et al. Screening a nation for hepatitis C virus elimination: a cross-sectional study
on prevalence of hepatitis C and associated risk factors in the Rwandan
general population. BMJ Open. 2019;9(7):e029743. The mobile clinic was funded by Partners in Health/Inshuti Mu Buzima, while
the government of Rwanda provided drugs. The data collection, analysis,
interpretation and writing the manuscript were conducted independently by
authors. 13. Umutesi G, Shumbusho F, Kateera F, Serumondo J, Kabahizi J, Musabeyezu
E, et al. Rwanda launches a 5-year national hepatitis C elimination plan: a
landmark in sub-Saharan Africa. J Hepatol. 2019;70(6):1043–5. Consent for publication
Not Applicable. 18. Harris PA, Taylor R, Thielke R, Payne J, Gonzalez N, Conde JG. Research
electronic data capture (REDCap)—a metadata-driven methodology and
workflow process for providing translational research informatics support. J
Biomed Inform. 2009;42(2):377–81. 18. Harris PA, Taylor R, Thielke R, Payne J, Gonzalez N, Conde JG. Research
electronic data capture (REDCap)—a metadata-driven methodology and
workflow process for providing translational research informatics support. J
Biomed Inform. 2009;42(2):377–81. Competing interests 19. Edejer TT-T, World Health Organization, editors. Making choices in health:
WHO guide to cost-effectiveness analysis. Geneva: World Health
Organization; 2003. 312 p. 19. Edejer TT-T, World Health Organization, editors. Making choices in health:
WHO guide to cost-effectiveness analysis. Geneva: World Health
Organization; 2003. 312 p. The authors declare that they have no competing interests. Authors’ contributions
h
l
h 8. Rao VB, Johari N, du Cros P, Messina J, Ford N, Cooke GS. Hepatitis C
seroprevalence and HIV co-infection in sub-Saharan Africa: a systematic
review and meta-analysis. Lancet Infect Dis. 2015;15(7):819–24. IK is the principal author of this manuscript in all steps from the conception
to the submission, data collection, analysis and manuscript writing. FN, JPG,
MU, EN, SW, CM, JDG, JDM, JS, FK and JDN contributed significantly in
design of the model and analysis presented in this paper. IK, DAB, FN, JPG,
MU carried out the data analysis and contributed to the development of the
tables. All authors participated in interpretation of the data. IK and DAB
conducted the literature review and wrote the first draft of the manuscript. JDN, JDM, SW, JS and FK reviewed and provided critique of the manuscript. The author(s) read and approved the final manuscript. 9. Han R, Zhou J, François C, Toumi M. Prevalence of hepatitis C infection
among the general population and high-risk groups in the EU/EEA: a
systematic review update. BMC Infect Dis. 2019;19(1):655. 10. Rosenberg ES, Rosenthal EM, Hall EW, Barker L, Hofmeister MG, Sullivan PS,
et al. Prevalence of hepatitis C virus infection in US states and the District of
Columbia, 2013 to 2016. JAMA Netw Open. 2018;1(8):e186371. 11. Makuza JD, Liu CY, Ntihabose CK, Dushimiyimana D, Umuraza S, Nisingizwe
MP, et al. Risk factors for viral hepatitis C infection in Rwanda: results from a
nationwide screening program. BMC Infect Dis. 2019;19(1):688. Ethics approval and consent to participate 15. Rwanda Biomedical Centre, Ministry of Health. Rwanda National HIV and Viral
Hepatitis Annual Report 2017-2018. Kigali: Rwanda Biomedical Centre; 2018. 15. Rwanda Biomedical Centre, Ministry of Health. Rwanda National HIV and Viral
Hepatitis Annual Report 2017-2018. Kigali: Rwanda Biomedical Centre; 2018. The approval of this study was obtained from Inshuti Mu Buzima Research
Committee (IMBRC) and Rwanda National Ethics Committee (RNEC) 015/
RNEC/2020). This study used retrospective data, which was collected as part
of routine clinical practice; therefore informed consent was not required by
the Rwanda National Ethics Committee. 16. Makuza JD. Training of health care providers on viral hepatitis prevention
and management. Musanze: Rwanda Biomedical Centre; 2019. 16. Makuza JD. Training of health care providers on viral hepatitis prevention
and management. Musanze: Rwanda Biomedical Centre; 2019. 17. Viana MSVB, Takei K, Collarile Yamaguti DC, Guz B, Strauss E. Use of AST
platelet ratio index (APRI score) as an alternative to liver biopsy for
treatment indication in chronic hepatitis C. Ann Hepatol. 2009;8(1):26–31. 17. Viana MSVB, Takei K, Collarile Yamaguti DC, Guz B, Strauss E. Use of AST
platelet ratio index (APRI score) as an alternative to liver biopsy for
treatment indication in chronic hepatitis C. Ann Hepatol. 2009;8(1):26–31. Author details
1 20. Ministry of Health, Rwanda, Mylan Laboratories Limited. Memorandum of
Understanding. 2019. 20. Ministry of Health, Rwanda, Mylan Laboratories Limited. Memorandum of
Understanding. 2019. 1Partners In Health/Inshuti Mu Buzima, Rwinkwavu, Rwanda. 2Department of
Global Health and Social Medicine, Harvard Medical School, Boston, USA. 3GlaxoSmithKline, Philadelphia, PA, USA. 4Rwanda Biomedical Centre, IHDPC,
Kigali, Rwanda. 5School of Population and Public Health, University of British
Columbia, Vancouver, Canada. 21. Chemouni B. The political path to universal health coverage: power, ideas and
community-based health insurance in Rwanda. World Dev. 2018;106:87–98. 21. Chemouni B. The political path to universal health coverage: power, ideas and
community-based health insurance in Rwanda. World Dev. 2018;106:87–98. 22. National Institute of Statistics of Rwanda (NISR). Rwanda Poverty Profile
Report ,2016/17. Kigali NISR; 2018. 22. National Institute of Statistics of Rwanda (NISR). Rwanda Poverty Profile
Report ,2016/17. Kigali NISR; 2018. 23. Hitimana R, Lindholm L, Krantz G, Nzayirambaho M, Pulkki-Brännström A-M. Cost of antenatal care for the health sector and for households in Rwanda. BMC Health Serv Res. 2018;18(1):262. 23. Hitimana R, Lindholm L, Krantz G, Nzayirambaho M, Pulkki-Brännström A-M. Cost of antenatal care for the health sector and for households in Rwanda. BMC Health Serv Res. 2018;18(1):262. Publisher’s Note Springer Nature remains neutral with regard to jurisdictional claims in
published maps and institutional affiliations.
|
W2742273901.txt
|
https://www.biogeosciences.net/14/3685/2017/bg-14-3685-2017.pdf
|
en
|
Reviews and syntheses: An empirical spatiotemporal description of the global surface–atmosphere carbon fluxes: opportunities and data limitations
|
Biogeosciences
| 2,017
|
cc-by
| 15,505
|
Biogeosciences, 14, 3685–3703, 2017
https://doi.org/10.5194/bg-14-3685-2017
© Author(s) 2017. This work is distributed under
the Creative Commons Attribution 3.0 License.
Reviews and syntheses: An empirical spatiotemporal description
of the global surface–atmosphere carbon fluxes:
opportunities and data limitations
Jakob Zscheischler1,2 , Miguel D. Mahecha2,3,4 , Valerio Avitabile5 , Leonardo Calle6 , Nuno Carvalhais2,7 ,
Philippe Ciais8 , Fabian Gans2 , Nicolas Gruber9 , Jens Hartmann10 , Martin Herold5 , Kazuhito Ichii11,12 , Martin Jung2 ,
Peter Landschützer9,13 , Goulven G. Laruelle14 , Ronny Lauerwald14,15 , Dario Papale16 , Philippe Peylin7 ,
Benjamin Poulter6,17 , Deepak Ray18 , Pierre Regnier14 , Christian Rödenbeck2 , Rosa M. Roman-Cuesta5 ,
Christopher Schwalm19 , Gianluca Tramontana16 , Alexandra Tyukavina20 , Riccardo Valentini21 , Guido van der
Werf22 , Tristram O. West23 , Julie E. Wolf23 , and Markus Reichstein2,3,4
1 Institute
for Atmospheric and Climate Science, ETH Zurich, Universitätstr. 16, 8092 Zurich, Switzerland
Planck Institute for Biogeochemistry, Hans-Knöll-Str. 10, 07745 Jena, Germany
3 German Centre for Integrative Biodiversity Research (iDiv), Deutscher Platz 5e, 04103 Leipzig, Germany
4 Michael Stifel Center Jena for Data-Driven and Simulation Science, 07743 Jena, Germany
5 Wageningen University & Research, Laboratory of Geo-Information Science and Remote Sensing, Droevendaalsesteeg 3,
6708 PB Wageningen, the Netherlands
6 Institute on Ecosystems and Department of Ecology, Montana State University, Bozeman, MT 59717, USA
7 CENSE, Departamento de Ciências e Engenharia do Ambiente, Faculdade de Ciências e Tecnologia,
Universidade NOVA de Lisboa, Caparica, Portugal
8 Laboratoire des Sciences du Climat et de l’Environnement, CEA-CNRS-UVSQ, 91191, Gif sur Yvette, France
9 Institute of Biogeochemistry and Pollutant Dynamics, ETH Zurich, Zurich, Switzerland
10 Institute for Geology, CEN – Center for Earth System Research and Sustainability, University of Hamburg, Hamburg,
Germany 55, 20146 Hamburg, Germany
11 Department of Environmental Geochemical Cycle Research, Agency for Marine-Earth Science and Technology,
Yokohama, Japan
12 Center for Global Environmental Research, National Institute for Environmental Studies, Tsukuba, Japan
13 Max Planck Institute for Meteorology, Bundesstr. 53, Hamburg, Germany
14 Dept. Geoscience, Environment & Society (DGES), CP160/02, Université Libre de Bruxelles, 1050 Brussels, Belgium
15 College of Engineering, Mathematics and Physical Sciences, University of Exeter, EX4 4QE Exeter, Devon, UK
16 Department for Innovation in Biological, Agro-food and Forest systems (DIBAF), University of Tuscia,
Viterbo, 01100, Italy
17 NASA Goddard Space Flight Center, Biospheric Sciences Laboratory, Greenbelt, MD 20771, USA
18 Institute on the Environment (IonE), University of Minnesota, Saint Paul, MN 55108, USA
19 Woods Hole Research Center, Falmouth MA 02540, USA
20 Department of Geographical Sciences, University of Maryland, College Park, MD, USA
21 CMCC, Via A. Imperatore, 16, 73100, Lecce, Italy
22 Faculty of Earth and Life Sciences, Vrije Universiteit Amsterdam, Amsterdam, the Netherlands
23 Joint Global Change Research Institute, Pacific Northwest National Laboratory, College Park, MD, USA
2 Max
Correspondence to: Jakob Zscheischler (jakob.zscheischler@env.ethz.ch) and Miguel D. Mahecha
(mmahecha@bgc-jena.mpg.de)
Received: 8 October 2016 – Discussion started: 20 October 2016
Revised: 6 June 2017 – Accepted: 28 June 2017 – Published: 9 August 2017
Published by Copernicus Publications on behalf of the European Geosciences Union.
3686
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
Abstract. Understanding the global carbon (C) cycle is of
crucial importance to map current and future climate dynamics relative to global environmental change. A full characterization of C cycling requires detailed information on spatiotemporal patterns of surface–atmosphere fluxes. However,
relevant C cycle observations are highly variable in their coverage and reporting standards. Especially problematic is the
lack of integration of the carbon dioxide (CO2 ) exchange of
the ocean, inland freshwaters and the land surface with the
atmosphere. Here we adopt a data-driven approach to synthesize a wide range of observation-based spatially explicit
surface–atmosphere CO2 fluxes from 2001 to 2010, to identify the state of today’s observational opportunities and data
limitations. The considered fluxes include net exchange of
open oceans, continental shelves, estuaries, rivers, and lakes,
as well as CO2 fluxes related to net ecosystem productivity, fire emissions, loss of tropical aboveground C, harvested
wood and crops, as well as fossil fuel and cement emissions.
Spatially explicit CO2 fluxes are obtained through geostatistical and/or remote-sensing-based upscaling, thereby minimizing biophysical or biogeochemical assumptions encoded
in process-based models. We estimate a bottom-up net C exchange (NCE) between the surface (land, ocean, and coastal
areas) and the atmosphere. Though we provide also global
estimates, the primary goal of this study is to identify key
uncertainties and observational shortcomings that need to be
prioritized in the expansion of in situ observatories. Uncertainties for NCE and its components are derived using resampling. In many regions, our NCE estimates agree well with
independent estimates from other sources such as processbased models and atmospheric inversions. This holds for Europe (mean ± 1 SD: 0.8 ± 0.1 PgC yr−1 , positive numbers are
sources to the atmosphere), Russia (0.1 ± 0.4 PgC yr−1 ), East
Asia (1.6 ± 0.3 PgC yr−1 ), South Asia (0.3 ± 0.1 PgC yr−1 ),
Australia (0.2 ± 0.3 PgC yr−1 ), and most of the Ocean regions. Our NCE estimates give a likely too large CO2 sink
in tropical areas such as the Amazon, Congo, and Indonesia. Overall, and because of the overestimated CO2 uptake
in tropical lands, our global bottom-up NCE amounts to a
net sink of −5.4 ± 2.0 PgC yr−1 . By contrast, the accurately
measured mean atmospheric growth rate of CO2 over 2001–
2010 indicates that the true value of NCE is a net CO2 source
of 4.3 ± 0.1 PgC yr−1 . This mismatch of nearly 10 PgC yr−1
highlights observational gaps and limitations of data-driven
models in tropical lands, but also in North America. Our
uncertainty assessment provides the basis for setting priority regions where to increase carbon observations in the future. High on the priority list are tropical land regions, which
suffer from a lack of in situ observations. Second, extensive pCO2 data are missing in the Southern Ocean. Third,
we lack observations that could enable seasonal estimates
of shelf, estuary, and inland water–atmosphere C exchange.
Our consistent derivation of data uncertainties could serve
Biogeosciences, 14, 3685–3703, 2017
as prior knowledge in multicriteria optimization such as the
Carbon Cycle Data Assimilation System (CCDAS) and atmospheric inversions, without over- or under-stating bottomup data credibility. In the future, NCE estimates of carbon
sinks could be aggregated at national scale to compare with
the official national inventories of CO2 fluxes in the land use,
land use change, and forestry sector, upon which future emission reductions are proposed.
1
Introduction
The global carbon (C) cycle is crucial for sustaining life on
Earth (Vernadsky, 1926). Humans have largely modified the
C cycle over centuries if not millennia (Ruddiman, 2003;
Pongratz et al., 2009). In the Industrial Era, the humancaused perturbation of the C cycle is largely driven by emissions of CO2 from burning fossil fuel C previously stored in
geological deposits, and changes in land use, which transfer
CO2 from C stocks in the land biosphere to the atmosphere,
but can also result into CO2 removal and increase of land
stocks. As those anthropogenic C emissions are partly taken
up by oceans and terrestrial ecosystems not affected by land
use change, the different reservoirs of the global C cycle and
the fluxes between them change over time (Houghton, 2007).
A precise knowledge of the various stocks and fluxes in the
C cycle is a prerequisite to monitor these changes and make
well-informed predictions under future climate change.
The Global Carbon Project (GCP) has made major efforts
in this direction and its annual updates of the global C budget have become a key source of information for the scientific
community and policy makers (Le Quéré et al., 2015). The
GCP annual C budget quantifies the partitioning of anthropogenic C emissions among the atmosphere, land, and ocean
components of the global C cycle, and separates the net land
flux into land use change emissions and a so-called “residual
land C sink” obtained by difference with other terms of the
budget and thus corresponding to the net land–atmosphere
CO2 flux over non-land-use affected ecosystems. The budget
of the GCP focuses on annual values integrated at the global
scale. An important point is that the GCP budget quantifies
solely the anthropogenic perturbation of CO2 fluxes, i.e. it
provides information about the fate of anthropogenic CO2
emissions in natural reservoirs (Ciais et al., 2013). According to the GCP, during the years 2000–2009 about 45 % of
the anthropogenic CO2 emissions each year stay in the atmosphere, the rest being taken up by the oceans (27 %) and land
(27 %) (Le Quéré et al., 2015).
Recently, a case has been made for a globally policyrelevant integrated C observation and analysis system (Ciais
et al., 2014). This system would go beyond the update of
global budgets, for which the CO2 growth rate accurately
measured at a single station (e.g. Mauna Loa) is sufficient
www.biogeosciences.net/14/3685/2017/
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
to constrain the global annual time–space integral of all
CO2 sources and sinks. It proposes to quantify regional CO2
fluxes with sufficient spatial details to monitor the effectiveness of CO2 mitigation and to detect and monitor trends of
CO2 losses and gains by land and terrestrial systems. This is
partly relevant for monitoring country-level intended nationally determined contributions (INDCs) to incept a CO2 emission trajectory consistent with global warming below 2 ◦ C
(UNFCCC, 2015). In such a policy-relevant C observing system, an uncertainty assessment for each data stream of CO2
fluxes at different spatial and temporal scales is important to,
for instance, identify significant regional emission hotspots
and trends in emissions and sinks (Ciais et al., 2014).
The steadily increasing number of Earth observations, in
particular since the start of the satellite era, has improved
our knowledge of the Earth system (Berger et al., 2012;
Tatem et al., 2008). Especially C cycle science has benefited from globally available satellite observations and community efforts to unify in situ observational networks such
as FLUXNET on land (Baldocchi, 2014), the Surface Ocean
CO2 Atlas (SOCAT) (Bakker et al., 2014), and more recently CO2 outgassing from lakes and rivers (Raymond et al.,
2013). Combining these available point measurements of either CO2 fluxes (e.g. from eddy covariance towers on land),
or variables that can be directly related to CO2 fluxes (e.g.
pCO2 over aquatic surfaces) with climate fields and remotely
sensed variables (e.g. vegetation greenness), provides a basis to robustly upscale surface–atmosphere CO2 exchange to
larger areas using statistical models (Jung et al., 2011; Rödenbeck et al., 2015).
In this study we aim at characterizing the ability of current C cycle observations on ground for quantifying a spatiotemporally explicit picture of the net CO2 exchange between the Earth’s surface (terrestrial and aquatic) and the atmosphere (NCE). Unlike the GCP global budget of anthropogenic CO2 , we consider here the full contemporary net
exchange of surface–atmosphere CO2 fluxes. We focus our
analysis on fluxes that can be directly derived from observations. That is, we use data-driven empirical models instead
of process-based models that are only indirectly constrained
by observations. Further, we only consider “bottom-up” estimates derived from measurements at the Earth’s surface or
from satellites. Inversions, which largely rely on atmospheric
measurements in combination with a transport model, are not
directly included but used for comparison. The goal of this
analysis is to test the up-scaling of local flux-related observations to regional and global budgets, and point out the limitations of the current observational networks and data-driven
models used to interpolate point-scale CO2 fluxes across
larger scales, for quantifying the most important CO2 fluxes
exchanged between the Earth’s surface and the atmosphere.
One of the major innovations of this study is combining
data-driven estimates of oceanic, inland waters, and terrestrial ecosystems CO2 exchange and providing spatially explicit maps of the CO2 exchange between the surface and the
www.biogeosciences.net/14/3685/2017/
3687
atmosphere at a monthly scale for the decade 2001–2010. At
the same time, by adding emissions from fossil fuels and cement production and comparing with the annual growth rate
of CO2 , we identify the limits of a C budget purely driven
by surface data. We characterize regions in which surface–
atmosphere CO2 fluxes are most uncertain based on the currently available data and the models used for upscaling, and
thus point out regions where either more observations or a
better understanding of the processes are necessary. It is not
the primary goal of this study to provide the best global CO2
flux inventory, but rather to identify the key uncertainties and
observational shortcomings that need to be prioritized in the
expansion of in situ observatories.
The paper is structured as follows. In Sect. 2 we introduce the different data streams used in the analysis, including
spatially explicit estimates of aquatic and terrestrial CO2 exchange. In Sect. 3 we present the resulting combined synthesis as global maps, regionally aggregated fluxes, absolute and
relative uncertainties, latitudinal averages and seasonal cycles. Section 4 addresses the benefits and limits of the current
observational system for constraining global net CO2 fluxes.
Section 5 summarizes the main conclusions drawn from this
synthesis.
2
Data and methods
We collected ensembles of data-driven estimates of the NCE
for the major subsystems of the Earth from 2001 to 2010
(Table 1). Each data set was aggregated to 1 × 1◦ spatial resolution. All data sets have an original spatial resolution of
at least 1 × 1◦ and were aggregated to the lower-resolution
common grid. The temporal resolution is monthly. For data
sets that were only available at yearly timescale or once
over the complete time period (Table 1), we distributed
fluxes evenly across all months. In this synthesis, we include NCE from open oceans, continental shelves, estuaries, rivers, lakes, and terrestrial ecosystems, which we combine with estimates of fossil fuel and cement emissions (FF).
The terrestrial ecosystem component accounts for fire emissions (Fire), loss of tropical above-ground biomass assumed
to be released as CO2 to the atmosphere (ELUC ), emissions
of the CO2 contained in harvested wood (Wood) and crops
(Crops), and net ecosystem productivity (NEP). We combine
fluxes from oceans, shelves, and estuaries into a homogeneous marine flux product in order to account for overlapping or missing regions from the different aquatic products
(Marine, Sect. 2.2.6). We further compare the net CO2 exchange (NCE) derived from the combination of all the above
products with the growth rate of atmospheric CO2 (CGR).
Combining all fluxes, the overall NCE between the Earth’s
surface and the atmosphere is given as
NCE = Marine + Lakes + Rivers − NEP
+ Crops + Wood + ELUC + Fire + FF.
(1)
Biogeosciences, 14, 3685–3703, 2017
3688
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
Table 1. Data sets used in this study including reference, time period and number of ensemble runs. If not specified, temporal resolution is
monthly.
Data set
Reference
Time period used
Ocean
Landschützer et al. (2014)
Rödenbeck et al. (2014)
Laruelle et al. (2014)
Laruelle et al. (2013)
2001–2010
Shelf
Estuaries
Marine
Rivers
Lakes
NEP
Crops
Wood
Fire
ELUC
FF (fossil fuels)
Atmospheric growth rate
Lauerwald et al. (2015)
Raymond et al. (2013)
Tramontana et al. (2016)
Wolf et al. (2015b)
Poulter (2015)
Giglio et al. (2013)
Tyukavina et al. (2015)
Harris et al. (2012)
CARBONES
NOAA
No. runs
5 + 5 = 10
1 estimate
1 estimate
2001–2010
1 estimate
1 estimate
2001–2010
2001–2010, annual
2000, 1 estimate
2001–2010
2000–2012, 1 estimate
2000–2005, 1 estimate
2001–2010
2001–2010
1
1
10
50
1
8
10
1
1
1+1 = 2
1
1
All units were transformed into fluxes of C per unit time.
If all CO2 fluxes were included, NCE would translate into
the CGR. By convention negative fluxes indicate an uptake by the Earth surface. Data scarcity precludes including
all known vertical CO2 fluxes in this study. Missing fluxes
include geological CO2 fluxes, erosion-related fluxes, nonCO2 fluxes, wood product pools decay, and biofuel burning.
the coefficient of variation (CV = IAV/mean) for each of the
nine flux terms in Eq. (1).
2.1
For the global open ocean flux estimate we used two complementary data-driven estimates (Table 1). Both approaches
computed maps of the sea surface partial pressure of CO2
(pCO2 ). They relied on the surface ocean CO2 observations
from the SOCATv2 database (Bakker et al., 2014) and filled
data gaps by either establishing relationships between auxiliary driver data and observations, which can then be applied to extrapolate pCO2 in regions without data coverage (SOM-FFN; Landschützer et al., 2014), or by assimilating the available observations in a mass-balance model of
the mixed layer and directly interpolating data gaps (Jena
CarboScope mixed-layer scheme oc_v1.2; Rödenbeck et al.,
2014). To test the established predictor–target relationship,
the SOM-FFN method holds back a certain fraction of the
observations proportional to the method’s degrees of freedom
for internal validation. Repeating this relationship building
process and withholding different sets of validation data has
created the five ensemble members used for this study. For
the Jena CarboScope mixed-layer scheme, we used the five
sensitivity cases with varied parameters such as changes in
correlation length as described in Rödenbeck et al. (2014).
The pCO2 fields of both methods have been validated
against independent observations (Landschützer et al., 2014,
2015; Rödenbeck et al., 2014) and were compared with other
complementary data-based interpolation methods (Rödenbeck et al., 2015), illustrating their good performance in reconstructing interannual variation.
Uncertainty estimation and propagation
For five of the nine variables contributing to the NCE of
Eq. (1), we obtained multiple spatiotemporally explicit estimates directly from the raw data products. These estimates
are expected to sample the uncertainty in each variable. We
could obtain 10 different estimates for the terms Marine and
Crops respectively, 50 estimates for Rivers, eight estimates
for NEP, two estimates for ELUC , and only one estimate
for each of the remaining four variables (Table 1). To estimate NCE including spatiotemporally explicit uncertainties,
we randomly draw ensemble members from each of these
nine terms to create an integrated ensemble for NCE (Eq. 1).
With the available estimates, we could in principle create
10×50×8×10×2 = 80 000 spatiotemporal explicit estimates
of NCE. From these 80 000 possible NCE estimates we randomly select 200 (to reduce computational expense) to construct the NCE ensemble, which is used in the remainder of
the paper. This resampling approach is illustrated in Fig. 1
and ensures a consistent propagation of spatiotemporally correlated uncertainties. For instance, by aggregating each member of NCE to the desired region and estimating uncertainty
through the 200 members, we can compute regional uncertainties. In addition, we computed mean fluxes, uncertainty
(defined as 1 standard deviation (SD) of the annual mean
across all realizations), interannual variability (IAV, defined
here as 1 SD of annual means across all available years) and
Biogeosciences, 14, 3685–3703, 2017
2.2
2.2.1
Aquatic fluxes
Oceans
www.biogeosciences.net/14/3685/2017/
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
3689
and open ocean waters using the shelf break as outer limit
(Laruelle et al., 2014).
2.2.3
Figure 1. Schematic explanation of the uncertainty propagation.
Each spatiotemporal estimate of NCE is computed as the sum of
randomly selected estimates of the nine fluxes contributing to NCE
(see Eq. 1, here denoted by Fi ). For this study we compute 200 estimates of NCE. Uncertainties can be assessed at different spatial
scales by first aggregating all NCE estimates to the desired scale
and then using the 200 members for uncertainty estimation.
The CO2 emissions from estuaries were derived from 161 annually averaged local CO2 air–water exchange rates reported
in the literature (Laruelle et al., 2013). The data were allocated to one of the 45 coastal MARCATS regions (MARgins and CATchments Segmentation) defined in Laruelle et
al. (2013) and further categorized among the four dominant estuarine types (i.e. small deltas, tidal systems, lagoons,
fjords; see Dürr et al., 2011) to calculate regionally averaged,
type-specific CO2 emission rates. In MARCATS regions devoid of estuarine data, the global average type-dependent air–
water CO2 flux was used from Laruelle et al. (2013). These
flux densities were then multiplied by the estuarine surface
areas for each type, estimated at 1◦ resolution from the length
of the coastline and a type-specific length to estuarine surface
ratio (Dürr et al., 2011).
2.2.4
Both methods calculate the air–sea flux using a bulk formulation of the air–sea CO2 transfer, driven by the air–
sea pCO2 difference (1pCO2 ) (Jähne et al., 1987) and
a quadratic dependence of the wind speed at a height of
10 m (Wanninkhof, 1992) updating the gas transfer coefficient to fit a mean transfer velocity of 16 cm per hour following Wanninkhof (2013). High-resolution wind speeds
at 10 m are calculated from the u and v wind components of the ERA-Interim wind speed analysis (Dee et
al., 2011) and atmospheric pCO2 fields, required to calculate the 1pCO2, are calculated are estimated from the
GLOBALVIEW-CO2 (2012) marine boundary layer CO2
product.
2.2.2
Estuaries
Marine
We created 10 marine estimates by combining the 10 estimates for the open oceans with shelves and estuaries to a
consistent marine product. For pixels with observations from
multiple products (e.g. estuaries and oceans) we follow a
“priority rule” whereby the shelves, estuaries, or oceans observation value only (in that order) is retained. Empty pixels
are gap-filled with a 3 × 3 mean window. This same filter is
also applied to the rest of the merged data set to smooth out
hard borders between the different estimates. This application does not significantly change the overall flux estimates,
but arguably results in a more realistic interface. Note that in
the merged Marine product, uncertainty and IAV could only
be assessed for the ocean flux.
Shelves
2.2.5
For continental shelf seas we derived the 1pCO2 from
3 × 106 surface pCO2 measurements extracted from the SOCATv2 database (Bakker et al., 2014) and observational atmospheric pCO2 data (GLOBALVIEW-CO2, 2012). The local CO2 air–sea flux values were then obtained using a winddependent quadratic formulation parameterized as in Wanninkhof et al. (2013) and wind speeds extracted from a crosscalibrated multiplatform (CCMP) high-resolution data product for ocean surface winds (Atlas et al., 2011). The resulting local fluxes were then integrated spatially over 150
coastal regions (COSCATs – COastal Segmentation and related CATchments; Laruelle et al., 2013; Meybeck et al.,
2006) using distinct integration methods depending on the
data density (Laruelle et al., 2014). In addition, a temporal
integration was also performed at the monthly, seasonal, or
yearly timescale depending on the data coverage. These temporally and regionally averaged air–sea CO2 fluxes were then
disaggregated using a 1◦ resolution map excluding land areas
www.biogeosciences.net/14/3685/2017/
Rivers
Fifty estimates of CO2 evasion from streams and rivers were
derived from a spatially explicit, empirical model of river
water pCO2 and global maps of stream surface areas and
gas exchange velocities at a resolution of 0.5◦ (Lauerwald et
al., 2015). The empirical pCO2 model was trained on 1182
river catchments from the GLORICH database (Hartmann et
al., 2014) for which averages of pCO2 could be calculated.
Steepness of terrain, terrestrial net primary production, average air temperature, as well as population density were identified as predictors (R 2 = 0.47). The global maps of stream
surface area and gas exchange velocities were obtained by
a GIS-based application of published empirical scaling laws
(Raymond et al., 2012, 2013) using topography (Lehner et
al., 2008) and runoff (Fekete et al., 2002). The CO2 evasion was calculated as product of water–air pCO2 gradient
(assuming an atmospheric pCO2 of 390 µatm), river surface
areas, and gas exchange velocities. A Monte Carlo simulaBiogeosciences, 14, 3685–3703, 2017
3690
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
tion based on standard errors of the predictors in the pCO2
model and uncertainty ranges for estimates of stream surface
area and gas exchange velocity was run to produce 50 CO2
evasion estimates.
2.2.6
Lakes
Estimates of CO2 evasion from lakes and reservoirs were
taken from Raymond et al. (2013), which reports average
lake pCO2 , total lake/reservoir surface area, and total CO2
evasion for 231 COSCAT regions (including endorheic regions). For the total lake/reservoirs surface area, data from
the Global Lakes and Wetland Database (GLWD; Lehner and
Döll, 2004) were combined with an estimate for small lakes
and reservoirs not represented in the GLWD using a scaling law. Here, we used the GLWD data to downscale the
estimates of Raymond et al. (2013) to a continuous 1◦ resolution. For this purpose, we combined a uniform air–water
CO2 flux (per unit surface area) within each COSCAT region with a spatially explicit estimate of the lakes/reservoirs
surface at this resolution. The small lakes/reservoirs not represented in the GLWD were assumed evenly distributed over
the COSCAT area.
2.3
2.3.1
Terrestrial fluxes
NEP
We used eight empirical, machine learning based products
from FLUXCOM (www.fluxcom.org) for net ecosystem productivity (NEP), derived from more than 200 FLUXNET
sites and exclusively remote-sensing-based predictor variables (“FLUXCOM-RS”; see Tramontana et al., 2016). The
eight machine learning methods used here include artificial
neural networks, four variants of model or regression tree
ensembles, kernel methods (support vector machines, kernel ridge regression), and multivariate adaptive regression
splines (Tramontana et al., 2016). All methods were trained
on 8-daily tower-based NEP estimates.
2.3.2
Crops
About 42 % of global crop biomass is harvested, transported,
and respired offsite (Wolf et al., 2015a). The impact of this
lateral C transport on fluxes can be seen at the country scale
in the form of import and exports, but even more so at subregional scales where the movement of crop biomass to feed
livestock and humans is evident (Hayes et al., 2012; West et
al., 2011). To capture the spatial distribution of CO2 fluxes
from agricultural harvest, we used livestock and human CO2
emissions estimates (Wolf et al., 2015b) that are available
from 2005 to 2011 at 0.05◦ spatial resolution. CO2 that has
previously been taken up from the atmosphere by the harvested biomass of crops is included in the NEP estimates
from FLUXCOM. We aggregated best estimates of the data
to 1◦ , added all uncertainty estimates within one 1◦ pixel and
Biogeosciences, 14, 3685–3703, 2017
used them as estimates for 1 standard deviation on the new
1◦ grid. Assuming Gaussian-distributed errors we sampled
1000 values at each pixel and used 10 maps of the 5th, 15th,
. . . , 95th quantiles as different ensemble members. Data were
then linearly extrapolated back to 2001–2004. In a final step,
and because it is not known in which months the emissions
occur, we further distributed the annual estimates equally
across all 12 months.
2.3.3
Wood
We used one estimate of globally gridded forest harvesting
data around year 2000 as described in the Supplement S1.
These data include fuelwood and roundwood harvested volumes in m3 . We translated wood volumes into units of C using a value of 0.275 MgC m−3 from FAO (http://www.fao.
org/docrep/w4095e/w4095e06.htm), assuming wood density
of 0.55 t m−3 . To avoid double counting wood harvest with
aboveground biomass loss in tropical areas exposed to land
use change, we use wood harvesting data only in locations
where the amount of harvested wood (in C) exceeds ELUC
(Sect. 2.3.4). We assume that 100 % of the harvested wood
is respired back to the atmosphere within a year, thus assuming no change in C stock of wood products and constant harvesting rates across years. However, C contained in harvested
wood is usually emitted at a different location than where the
harvest took place. We thus incorporated lateral shifts of harvested wood by redistributing wood harvest according to the
consumption of wood as explained in the Supplement (see
also Fig. S2).
2.3.4
ELUC
We used two estimates for CO2 fluxes due to tropical deforestation and degradation. It is assumed here that 100 %
of biomass loss is converted to a CO2 flux being released instantly (within a year) to the atmosphere. In reality, a fraction
of lost tropical biomass decays in ecosystems (belowground
biomass and slash) and a fraction is used in wood products
of various lifetime. However, slash is decomposed fast and
biomass from deforested areas is transformed on average to
short-lived products (≈ 5 years after Earles et al., 2012).
1. Gross tropical deforestation emissions were taken from
Harris et al. (2012). They represent total (above- and belowground) C loss from gross forest cover loss in the
tropical regions due to human or natural causes (e.g.
disturbances without forest recovery) for the period of
2000–2005.
2. More recent estimates of aboveground C loss in the
tropics from stand-replacement disturbance of forest
cover due to human or natural causes were provided
by Tyukavina et al. (2015). Sample-based estimates of
mean 2000–2012 aboveground C loss for each 30 m resolution forest C stratum were attributed to all pixels of
www.biogeosciences.net/14/3685/2017/
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
3691
Figure 2. Different components of observation-driven C exchange between the Earth’s surface and the atmosphere. Red arrows denote a
flux from the surface to the atmosphere (net source), green arrows denote a flux from the atmosphere to the surface (net sink). Units are in
PgC yr−1 .
the corresponding stratum and averaged to the 1 × 1◦
resolution.
We used ELUC only in those pixels where the average
of the two estimates (1 and 2) exceeds wood harvesting
(Sect. 2.3.3).
2.3.5
Fire
We used fire emissions from the Global Fire Emissions
Database version 4 with small fires (GFED4s, http://www.
globalfiredata.org) based on burned area from Giglio et
al. (2013) and Randerson et al. (2012) and an updated version of the biogeochemical modelling framework of van der
Werf et al. (2010) to convert burned area to C emissions.
We included all fire types except tropical deforestation and
degradation fires, which are included in ELUC and should
thus not be counted twice (Sect. 2.3.4). For an earlier version of fire emissions (GFED3) a Monte Carlo simulation
indicated an uncertainty of about 20 % (1SD) for continentalscale estimates but these estimates turned out to be not very
reliable (van der Werf et al., 2017). For example, the inclusion of small fire burned area led to an increase in burned
area exceeding the previously assumed uncertainty and the
current version therefore has no uncertainty assessment at
pixel level. Note that GFED fire emissions depend on estimates of net primary production and combustion factors as
computed by the CASA model.
2.3.6
FF
We use the IER-EDGARv4.2 product for fossil fuel and cement emissions, which was derived within the CARBONES
project by the Institute für Energiewirtschaft und Rationelle
Energieanwendung (IER). It is based on the Edgar v4.2 fossil fuel spatial distribution (with the highest spatial resolution of 0.1 × 0.1◦ ) and uses national consumption and global
www.biogeosciences.net/14/3685/2017/
production statistics. Based on the sectorial distinguished
EDGARv4.2 emissions, sector-specific and country-specific
temporal profiles were included. A detailed description of the
construction of the product is given at http://www.carbones.
eu/wcmqs/project/ccdas/#Fossil_Fuel. It is important to note
that FF emissions here are not observation based as the IEREDGARv4.2 product is partly based on national estimates
from official inventories reported by countries to the UNFCCC.
2.4
Atmospheric growth rate
We used the atmospheric rate of change of CO2 , which is
equal to the space and time integral of all emissions and
sinks at the surface, using the calculations made by the GCP
(Le Quéré et al., 2015). These calculations are based on
the global growth rate of atmospheric CO2 (CGR) provided
by the US National Oceanic and Atmospheric Administration Earth System Research Laboratory (NOAA/ESRL) and
were derived from multiple stations selected from the marine boundary layer sites with well mixed background air
(Ballantyne et al., 2012; Masarie and Tans, 1995). They
applied conversion from concentrations to carbon mass is
1 ppm = 2.12 PgC (Prather et al., 2012).
2.5
Inversions
For a comparison of yearly variability, spatial patterns, and
latitudinal bands of NCE, we used annual means of 10 atmospheric CO2 inversions collected in Peylin et al. (2013),
available at the same spatial and temporal resolution. Atmospheric CO2 inversions estimate the spatiotemporal explicit
CO2 exchange between the Earth surface and atmosphere using atmospheric CO2 measurements and a transport model.
Inversions have a closed budget by construction. The median and interquartile range for each year is taken over all
Biogeosciences, 14, 3685–3703, 2017
3692
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
Table 2. Net carbon exchange for different subsystems and variables that contribute to NCE (Eq. 1; negative numbers are surface uptake).
Uncertainty (Unc.) is SD over ensemble runs. IAV is SD over annual values (2001–2010) and CV is coefficient of variation, computed as
IAV divided by the mean.
Rivers
Lakes
−NEP
Crops
Wood
ELUC
FF
Fire
NCE
−1.60
0.15
0.36
0.22
0.65
0.08
0.32
−18.41
2.08
0.36
0.02
2.68
0.21
0.09
0.03
0.71
0.83
0.16
7.78
1.81
−5.45
1.99
0.75
0.10
0.11
0.06
45
90
(a)
0
available inversions for that year, as not all inversions were
available until 2010. Atmospheric CO2 inversions estimate
surface CO2 fluxes such that they best fit observed atmospheric constraints. They usually rely on prior information
provided by terrestrial and oceanic biogeochemical models
but are mostly independent from the bottom-up data sets included in the present synthesis. They further use FF as an
input and then provide the surface–atmosphere flux excluding FF.
gC m−2 yr−1
−700
−300
−100
100
300
500
−50 0
50
gC m−2 yr−1
700
45
90
(b)
−45
−90
gC m−2 yr−1
0
100
0
200
300
45
90
(c)
10
20
gC m−2 yr−1
0
Mean fluxes, their uncertainties, interannual variability
(IAV), and CV (the mean-normalized IAV) for all individual
fluxes contributing to NCE are presented in Table 2. Mean
fluxes are also summarized graphically in Fig. 2 (mean over
2001–2010). Our best surface-data-driven bottom-up global
estimate of NCE is −5.4 ± 2.0 PgC yr−1 . That means that the
observation-based data sets suggest a large net sink, even if
FF and ELUC are included in NCE. By contrast, the accurately measured CO2 growth rate constrains NCE to being
a net CO2 source to the atmosphere of 4.3 ± 0.1 PgC yr−1
(2001–2010, Le Quéré et al., 2015). Thus, there is a large
mismatch with our NCE of 9.7 ± 2.0 PgC yr−1 . This highlights that our observation-based NCE is biased towards a too
large sink. Potential reasons for this mismatch are discussed
in Sect. 4. For most fluxes, uncertainty estimates strongly
exceed IAV (Table 2). Interestingly, process-based models,
which are only indirectly constrained by observations, provide an NCE that matches roughly the CO2 growth rate
(Le Quéré et al., 2015). Developers of process-based models have access to CO2 growth rate data and may be in the
position to tune their models so that they give realistic NCE
values (Schwalm et al., 2015), whereas in our bottom-up approach, we conducted a blind up-scaling of ground measurements without trying to match the CO2 growth rate.
0
Global net carbon exchange
−45
3.1
Results
−500
−90
3
0.11
−45
Mean
Unc.
IAV
CV
Marine
−90
Variable
%
0
50
100
0
150
200
100
%
Figure 3. Gridded spatial patterns of NCE. (a) 2001–2010 decadal
mean. (b) Uncertainty; 1SD across the NCE ensemble. (c) Relative
uncertainty; uncertainty normalized by absolute mean. Latitudinal
plots in (b) and (c) denote median across latitudes.
3.2
Spatial patterns of net carbon exchange
The 200-member NCE ensemble and the uncertainty distribution of each flux component enables us to provide a best
estimate for a gridded average surface–atmosphere CO2 flux
map for the time period 2001–2010 (Fig. 3a). According to
these estimates, tropical land areas are a larger CO2 sink than
Biogeosciences, 14, 3685–3703, 2017
www.biogeosciences.net/14/3685/2017/
www.biogeosciences.net/14/3685/2017/
NCE
(b)
Terrestrial NCE
(c)
Aquatic NCE
0.6
−0.8
PgC yr -1 2°
−0.2 0.2 0.6
−0.8
−0.2 0.2
0.6
−0.8
−0.2 0.2
(a)
3693
(d)
−0.2 0.2
−0.8
the mid-latitudes despite the visible forest bands in North
America and Russia that function as sinks. In contrast, the
high latitudes indicate a relatively small source. In the ocean,
these patterns are reversed, with sources in the tropics and a
sink in the mid-latitudes. Clearly, there is a strong land–sea
contrast and land NCE is much higher in magnitude compared to ocean NCE. In areas with high human population
densities and active industry (Europe, eastern China, US,
South Africa), emissions from fossil fuels and cement production clearly dominate the land CO2 fluxes.
Absolute uncertainty of NCE generally scales with the
mean flux and is highest in the most productive areas over
land (Amazon basin, Congo basin, Indonesia; Fig. 3b). Due
to the small contribution of the oceans, absolute uncertainties are barely discernible there. Although gross air–sea exchange fluxes have typical uncertainties of more than 20 %,
their differences are determined from independent measurements with a much higher accuracy (Ciais et al., 2013).
Relative uncertainties, however, show very distinct patterns (Fig. 3c). These are high on land in semi-arid and arid,
and in mountainous regions (i.e. rather unproductive areas
with near-zero mean) such as Australia, the Middle East, the
Midwest US, the Sahel, South Africa, the Andes, and around
the Tibetan Plateau. Marine–atmosphere CO2 exchange is
most uncertain in relative terms in the Bay of Bengal and
in the Southern Ocean, which is known to be undersampled,
and where the two data-driven NCE fluxes show substantial
regional patterns (Landschützer et al., 2014; Rödenbeck et
al., 2014). In addition, linear features with high relative uncertainty are visible, especially in the Southern Hemisphere.
These are related to the borders of the clusters used for deriving homogeneous regions of sea–air exchange in one of
the ocean-exchange products, which result in this product
in strong spatial gradients in the sea surface pCO2 (Landschützer et al., 2014). Relative uncertainties are mostly below
100 % for the median across latitudinal bands (Fig. 3c). Only
in the Southern Ocean is the relative uncertainty substantially
higher, reflecting difficulties in reconstructing seasonal to interannual variabilities with sparse observational constraints
(Landschützer et al., 2014; Rödenbeck et al., 2014). Nevertheless, Landschützer et al. (2015) have shown that there is
a better agreement between the estimates of Landschützer et
al. (2014) and Rödenbeck et al. (2014) when low-frequency
variability, such as decadal variability, is analysed.
Averaged over latitudinal bands, the tropics are clearly
a CO2 sink (Fig. 4a), a feature of the FLUXCOM models
used for NEP, whereas mid-latitudes form a net CO2 source,
mostly due to fossil fuel and cement emissions surpassing
natural CO2 sinks. This latitudinal pattern is strongly driven
by the terrestrial fluxes (Fig. 4b). Marine and land aquatic
CO2 exchange in turn is about 4 times smaller in magnitude
and shows CO2 sources in the tropics and CO2 sinks in the
extratropics (Fig. 4c). The aquatic CO2 source in the tropics
is not only the result of the ocean air–sea exchange, but also
of the very intense river outgassing in low-latitude regions
0.6
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
NCE−FF
Inversion NCE
−50
0
Latitude
50
Figure 4. Median and interquartile range of different subsets of
NCE (2001–2010 decadal mean). (a) All fluxes. (b) Terrestrial
fluxes. (c) Aquatic fluxes. (d) NCE without fossil fuels from this
synthesis (black) and from inversions (blue).
(Lauerwald et al., 2015). NCE in the mid-latitudes is dominated by fossil fuel emissions (black line in Fig. 4d shows
NCE-FF). FF contribute little in the tropics and the high latitudes but offset land and ocean CO2 sinks in the northern
mid-latitudes so that the net CO2 balance of this latitude band
is a net CO2 source.
We use the land cover map of FLUXCOM to identify tropical forests (all pixels where broadleaved evergreen trees dominate). Tropical forest, which covers about
3.5 % of the Earth’s surface, are allocated a CO2 sink
of −5.0 ± 0.6 PgC yr−1 , which is unrealistic, if compared
to, for example, forest biomass inventories (Pan et al.,
2011). Without this large sink, global NCE would be
of −0.4 ± 1.8 PgC yr−1 . This corrected estimate (assuming neutral C exchange in tropical forests) is still a
sink more than 4 PgC yr−1 larger than the global NCE
accurately constrained by CO2 growth rate observations
(4.3 ± 0.1 PgC yr−1 ). Including missing fluxes (e.g. biogenic
fluxes and emissions from wetlands; see Sect. 4.2) for which
we do not have spatially explicit estimates (see Sect. 4.4)
could close this gap. These considerations suggest that the
CO2 sink of tropical forests from FLUXCOM is probably
strongly overestimated and responsible for at least half of
the global mismatch with the observed CO2 growth rate (see
Sect. 4.1 for further discussion).
Biogeosciences, 14, 3685–3703, 2017
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
2
3694
0
Land release
−4
Pg C yr - 1
−2
Land uptake
NCE
NCE − FF
Ciais et al. NCE
NA
SA
EU
AF
RU
EA
SAs
SEA
AU
Rest Ocean
Figure 5. NCE (2001–2010 decadal mean) in RECCAP regions
over land, including (red) and without fossil fuels (blue). Shown
are median, interquartile range (box), and 1.5× interquartile range
(whiskers). The regional estimates including uncertainties of NCE
collected in Ciais et al. (2017) are underlain in grey. NA: North
America; SA: South America; EU: Europe; AF: Africa; RU: Russia; EA: East Asia; SAs: South Asia; SEA: Southeast Asia; AU:
Australia; Rest: remaining land areas.
3.3
3.3.1
Net carbon exchange over the RECCAP regions
3.3.1 RECCAP over land
Here we compare our NCE estimates over land with largely
independent estimates of net ecosystem exchange (NEE)
over continental-scale regions collected in RECCAP (REgional Carbon Cycle Assessment and Processes). Ciais et
al. (2017) compiled regional estimates of land–atmosphere
CO2 fluxes in the RECCAP regions from the original publications, complemented by river CO2 outgassing fluxes from
Raymond et al. (2013) and Lauerwald et al. (2015). Thus,
these estimates are not fully independent of those presented
in this study because they use the same river fluxes, fire emissions, and FF emission. All other fluxes are independent. The
RECCAP budgets were based on inventories, and in some instances on process models results. For ELUC , RECCAP publications used regional data sets or bookkeeping models (a
bookkeeping model combines local or regional observationbased estimates of C stocks before land use change and trajectories of C stocks after land use change including slash,
biomass, soil carbon, and harvested wood products with
changing areas of different land use types, Houghton and
Nassikas, 2017), that are independent from estimates gathered in Sect. 2.3.4. The RECCAP regions include North
America (NA; King et al., 2015), South America (SA; Gloor
et al., 2012), Europe (EU; Luyssaert et al., 2012), Africa (AF;
Valentini et al., 2014), Russia (RU; Dolman et al., 2012),
East Asia (EA; Piao et al., 2012), South Asia (SAs; Patra
et al., 2013), and Australia (AU; Haverd et al., 2013). No
regional study is yet available for Southeast Asia (SEA).
Greenland, the Middle East, Ukraine, Kazakhstan and New
Zealand are omitted in regional RECCAP studies because of
Biogeosciences, 14, 3685–3703, 2017
the difficulty of obtaining local ground-based observations.
Ciais et al. (2017) collected the regional estimates and combined them with estimates of lateral transport to estimate C
budgets for each region. NEE in Ciais et al. (2017) minus
C export by rivers should in principle be equal to our NCE
estimates without FF over the same regions (Fig. 5). In regions without tropical forest except NA (that is, EU, RU, EA,
SAs, and AU) the estimates by Ciais et al. (2017) are within
the uncertainty range of our assessment. For NA and regions
containing the tropics, our approach shows a much stronger
C sink.
Using our methodology, the annual NCE-FF for all RECCAP regions amounts to −11.0 ± 1.9 PgC yr−1 in contrast
to −1.3 ± 0.6 PgC yr−1 in Ciais et al. (2017). If we exclude SA, AF, and SEA, the numbers are −2.8 ± 1.0 and
−1.5 ± 0.4 PgC yr−1 , respectively, bringing both estimates
in each other’s uncertainty range. For SA, AF and SEA,
the two estimates even differ in sign. While our estimates
indicate strong C sinks of −4.3 ± 0.5, −2.7 ± 0.9, and
−1.2 ± 0.3 PgC yr−1 , respectively, Ciais et al. (2017) report
0.1 ± 0.3, 0.1 ± 0.3, and 0.0 ± 0.2 PgC yr−1 .
Given that Ciais et al. (2017) rely on an independent
method, this demonstrates that a relatively good understanding and observational coverage of net C fluxes exists for EU,
RU, AU, SAs, and EA to some extent. It is somewhat surprising that both approaches largely differ over North America, where good observational coverage for instance through
eddy covariance towers exist. The comparison also reveals
the high uncertainties and biases in bottom-up estimates of
NCE over tropical forests (see Sect. 3.2, but also Gloor et al.,
2012; Valentini et al., 2014) and underlines the importance of
long-term ground-based measurement campaigns in those regions (e.g. RAINFOR, http://www.rainfor.org/: Malhi et al.,
2002; ATTO: Andreae et al., 2015; Zhou et al., 2014).
3.3.2
RECCAP over ocean
We compare our estimates of marine C exchange over the
ocean with estimates from the RECCAP initiative. The Pacific Ocean is divided into North Pacific extratropics (NP),
tropical pacific (TP), and South Pacific extratropics (SP)
(Ishii et al., 2014). The Atlantic Ocean is divided into Arctic Ocean (AR), northern subtropics (NS), equatorial (EQ),
and southern subtropics (SS) (Schuster et al., 2013). Further,
there are estimates for the northern (NI) and southern Indian
Ocean (SI) (Sarma et al., 2013) as well as for the Southern
Ocean (SO) (Lenton et al., 2013). While there is large overlap between the pCO2 data used in the RECCAP estimates
and our NCE estimates over oceans, different independent
methods have been used to obtain flux estimates of ocean
CO2 exchange (Sect. 2.2.1). Overall, the estimates from both
sources agree very well (Fig. 6) and show ocean net C release in tropical regions (TP, EQ, and NI) and net C uptake in
all other regions. In SO our estimates predict a smaller sink
compared to the RECCAP estimates, a difference probably
www.biogeosciences.net/14/3685/2017/
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
Atlantic Ocean
Indian Ocean
PgC month-1
0
Ocean release
−1
Ocean uptake
−0.6
Marine (this study)
RECCAP
NP
TP
SP
AR
NS
EQ
SS
J
NI
SI
SO
−2
Pg C year -1
0
2
4
Figure 6. Marine NCE in RECCAP regions over the ocean. Shown
are median, interquartile range (box), and 1.5× interquartile range
(whiskers) of the Marine fluxes. The RECCAP estimates including uncertainties are underlain in grey. NP: North Pacific extratropics; TP: tropical pacific; SP: South Pacific extratropics; AR: Arctic
Ocean; NS: northern subtropics; EQ: equatorial; SS: southern subtropics; NI: northern Indian Ocean; SI: southern Indian Ocean; SO:
Southern Ocean.
−4
NH NCE
NH inversions
SH NCE
SH inversions
−2
−0.4
Pg C yr - 1
−0.2 0.0
0.2
0.4
1
0.6
Pacific Ocean
3695
NCE−FF
Inversion NCE
2002
2004
2006
2008
2010
Figure 7. Comparison of NCE-FF with NCE from inversions (by
construction without FF) on interannual timescales. Both time series were zero-centred by adding an offset of 13.23 PgC yr−1 for
NCE-FF and 3.74 PgC yr−1 for NCE from inversions. Lines show
median, shading denotes interquartile range.
owing to the weak observational constraint (Landschützer
et al., 2014; Rödenbeck et al., 2014). Our estimates generally have smaller uncertainty ranges, which is because (i) the
RECCAP studies include many more approaches (including
process-based models, atmospheric and ocean inversions) in
their estimates and (ii) in our analysis we include the uncertainty from the ocean pCO2 products and their realizations
but do not account for the uncertainty in the kinetic gas transfer.
www.biogeosciences.net/14/3685/2017/
F
M
A
M
J
J
A
S
O
N
D
Figure 8. NCE median seasonal cycle and interquartile range for
Northern Hemisphere (NH, 0–90◦ N, blue) and Southern Hemisphere (SH, 90–0◦ S, green) for estimates from this study (dark
colours) and inversions (light colours).
3.4
Comparison with inversions
We compare NCE without FF (NCE-FF) with annual values
from 10 inversions estimating the surface–atmosphere CO2
flux without FF (Peylin et al., 2013). While both estimates
agree well in the mid-latitudes, they show opposite patterns
in the tropics and northern high latitudes (Fig. 4d). The estimates of NEP in our NCE-FF probably have a substantial
bias towards too much uptake over tropical land (Sect. 4.1).
The comparison suggests that CO2 fluxes are comparably
well constrained in the mid-latitudes where bottom-up and
top-down approaches agree. Similar results have been obtained in a comparison of a bottom-up upscaling approach
with a more recent inversion based on CO2 concentration data from the Greenhouse gases Observing SATellite
(GOSAT; Kondo et al., 2015). The temporal evolution between both estimates show little agreement except the trend
towards more net C uptake by the Earth’s surface (Fig. 7).
The comparison shows that NCE-FF estimated from this
study has lower interannual variability compared to inversion
estimates. Uncertainties are very high for our NCE-FF. In addition, the mean annual C uptake in our estimates is nearly
10 PgC yr−1 higher than for inversions.
3.5
Monthly variability and mean seasonal cycle
NCE in the Northern Hemisphere (NH) exhibits a much
stronger mean seasonal cycle compared to the Southern
Hemisphere (SH), ranging from a net C uptake of nearly
2 PgC (per month) in July to a net C release of about 0.9 PgC
in December and January (Fig. 8). The SH is always a net
C sink, ranging between slightly under 0.8 PgC uptake in
January to roughly 0.1 PgC in August and September. This
illustrates the “breathing of the Earth” – that is, vegetation
activity largely follows the annual cycle of the sun. NH NCE
is strongly offset by fossil fuel emissions. The uncertainties
Biogeosciences, 14, 3685–3703, 2017
3696
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
for the SH seasonal cycle are generally much lower than for
the NH fluxes due to the larger contribution of the latter to
the overall flux pattern, which is related to the distribution
of land areas. If compared to inversions, we find that both
estimates only match in the summer of the NH. In all other
months and in the SH, our NCE estimates show a consistently
much larger surface C sink. In addition, the NCE estimates
from this synthesis show a smaller amplitude of the mean
seasonal cycle compared to the inversions. The difference in
amplitude of the mean seasonal cycle is on average 0.7 PgC
for the NH and 0.4 PgC for the SH.
4
Current limitations of a bottom-up spatiotemporal
assessment of net carbon exchange
Our study shows that today’s spatiotemporally explicit
and independent bottom-up observation-driven estimates of
surface–atmosphere CO2 exchange suffer from large bias,
such that they do not match the global NCE well constrained
from the CO2 growth rate. This statement is not downgrading the advances in the area, but rather a systematic reflection of the state of current research and monitoring. In fact,
at the regional scale, those estimates are often well constrained and may be used for model–data integration studies and validation purposes. The regions where observationdriven CO2 exchange is constrained the best include Europe,
Russia, South Asia, East Asia, Australia and all oceanic regions except the Southern Ocean. The most likely candidate
for inducing the mismatch between data-driven estimates and
the atmospheric CO2 growth rate is terrestrial NEP. In particular, tropical NEP estimates suggest a too large tropical sink.
In the following sections, we discuss (i) the possible reasons
for the large bias in NEP (Sect. 4.1), (ii) which fluxes are
missing in our synthesis (Sect. 4.2), (iii) how this synthesis data set could be used for model–data fusion (Sect. 4.3),
(iv) uncertainties in fire emissions (Sect. 4.4), and (v) the
impact of missing seasonal cycles in some of the data sets
(Sect. 4.5).
4.1
Difficulties in estimating NEP over land
Correctly predicting NEP from remote sensing requires establishing universal relationships between those predictors
and respiratory processes (Jägermeyr et al., 2014; Tramontana et al., 2016). However, predicting such processes still
poses major challenges to researchers (Trumbore, 2006). The
CO2 flux related to heterotrophic decomposition processes,
for instance, relates to factors controlling biological activity
via temperature, moisture availability, and the decomposable
substrate material. The question how soil respiration or total ecosystem respiration depends on these variables is not
yet entirely understood. Advancing our knowledge on these
processes is challenging due to both a lack of theory of resBiogeosciences, 14, 3685–3703, 2017
piration and the difficulty of obtaining relevant data to test
models (Trumbore, 2006).
In addition to a good theory for respiration, information
on disturbance history (e.g. time since last fire) and forest
age would improve the upscaling of NEP from sites to regions (Ciais et al., 2014). Disturbances that cause physical
damage to vegetation properties tend to temporarily increase
respiration and reduce photosynthesis and thus alter the balance between gross C uptake and release. Disturbed ecosystems are thus initially assumed to be strong C sources until
plant production recovers. However, how these regrowth processes compensate a given disturbance regime cannot yet be
quantified at global scales, as the area covered by disturbed
ecosystems is variable and unknown (Ciais et al., 2014). For
example, regrowth of vegetation after fires and other disturbances is not well sampled neither in the FLUXNET stations
nor in the set of predictors used by the FLUXCOM models
and is assumed to be implicit in our NEP estimate. Furthermore, management can have strong effects on annual NEP of
croplands, which form large parts of the land surface (Jung
et al., 2011). However, not all of the relevant predictors (i.e.
disturbance maps, management practices, soil moisture) are
currently available to be included in empirical upscaling exercises (Tramontana et al., 2016).
In addition to the above difficulties, some regions are undersampled by eddy covariance towers and thus NEP is not
well constrained. This is the case for tropical forests and the
northern high latitudes. In the tropics, undersampling leads
to a large overestimation of net CO2 uptake in comparison
to inversion and forest inventories (Peylin et al., 2013; Pan et
al., 2011), whereas in the high latitudes it leads to a comparably large CO2 release (Fig. 4).
Given the difference between NCE and inversions in the
tropics (Fig. 4), we can assume that a bias of FLUXCOM
NEP towards a too high C sink is the main reason why the
C budget is not closed in our approach. This raises the question why upscaled NEP has such a strong systematic bias towards a sink, particularly in the tropics (see also Jung et al.,
2011). We suspect that the eddy covariance towers collected
in FLUXNET, which provide the empirical basis for the
global data-driven estimates (see Sect. 2.3.1) do not represent
the different age classes of forests very well. For instance,
young and regrowing forests with a generally higher-thanaverage NEP are possibly over-represented in FLUXNET.
However, such an age dependency (Amiro et al., 2010; Coursolle et al., 2012; Hyvönen et al., 2007; Magnani et al., 2007)
has not yet been included in global upscaling of NEP. This
hypothesis should be tested in future upscaling exercises.
4.2
Missing fluxes
Due to a focus on spatially explicit maps, not all known
fluxes between the land surface and the atmosphere are considered in our analysis. We assume that including the following fluxes may have an influence on the regional and global
www.biogeosciences.net/14/3685/2017/
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
flux estimates (estimates of the flux magnitude are given if
known):
– emissions from biogenic volatile organic compounds
(VOCs) amount to approximately 0.76 PgC yr−1 globally (Sindelarova et al., 2014);
– CO2 emissions from wetlands, estimated globally at
around 2.1 PgC yr−1 (Aufdenkampe et al., 2011);
– CH4 emissions from biogenic sources and animals;
– crop residues burning in households;
– biofuel burning;
– changes in land management, e.g. shifts in agriculture,
soil tillage, grassland ploughing and grazing;
– geological fluxes;
– Raymond et al. (2013) estimate a much higher river evasion (1.8 PgC yr−1 instead of 0.65 PgC yr−1 used in this
study).
All known missing fluxes add up to an additional C release of
about 4 PgC yr−1 . Although substantial, they do not cover the
mismatch of more than 9 PgC yr−1 by far (Sect. 3.1). However, they would suffice to close the budget if tropical forests
are assumed to be C neutral (tropical forests are responsible
for a net C sink of about 5 PgC yr−1 , Sect. 3.2). This significant amount of missing fluxes prohibits constraining FLUXCOM runs with all the remaining fluxes. In other words, we
cannot be certain of the bias in upscaled NEP as long as the
major fluxes are not quantified in a spatially explicit manner. Emissions from VOCs and wetlands should thus receive
particular attention if a consistent spatiotemporal picture of
vertical CO2 exchange is to be obtained.
4.3
Uncertainty estimates and model–data fusion
Our uncertainty estimates of ocean and land C exchange
likely underestimate the true uncertainty. In particular, Landschützer et al. (2014) estimated that the choice of sea–air
gas transfer formulation (also including other relationships
than quadratic) and the pCO2 mapping mismatch lead to an
uncertainty of 37 % for the global average over sea–air exchange between 1998 and 2011, with the majority of this uncertainty stemming from the gas transfer formulation. Furthermore, the uncertainty of NEP is likely underestimated
because all upscaling methods in FLUXCOM use the same
set of predictors (Tramontana et al., 2016). Hence, the uncertainty estimates only cover the uncertainty related to the
upscaling method but do not contain uncertainties related to
the selection of predictors or observational uncertainty of the
predictors themselves.
A comprehensive spatiotemporally explicit bottom-up estimate of NCE can be a powerful ingredient for model–data
www.biogeosciences.net/14/3685/2017/
3697
integration exercises (Rayner et al., 2005). Yet model–data
integration requires uncertainty characteristics of all data
streams used (Raupach et al., 2005). Furthermore, it is important that uncertainties can be described in terms of random errors (Ciais et al., 2014). Error estimates at the local
or regional level are difficult to use if no spatial error covariance matrix is available. The uncertainty analysis presented
in this study obtained through Monte Carlo sampling aims to
be of use for model–data integration studies. Errors are automatically propagated through different spatial resolutions
by aggregating the individual ensembles of NCE. Naturally,
efforts should be made to obtain error estimates for all integrated data sets (i.e. Wood, Fires, Shelves, Estuaries, and
Lakes). Nevertheless, this first integrated NCE estimate offers new possibilities for approaches such as the Carbon Cycle Data Assimilation System (CCDAS; Rayner et al., 2005),
by providing not only a full spatiotemporal grid of fluxes but
also a transparent and consistent error propagation scheme.
This can have also practical applications, for instance for designing new measurement campaigns in regions with high
uncertainties to reduce knowledge gaps in the global CO2
fluxes.
4.4
Uncertainties in fire emissions
Fire emission estimates combine satellite-based fire data with
ecosystems models. Uncertainties in global fire emission estimates are substantial and different fire products vary largely
by location, vegetation type, and fire weather (Ciais et al.,
2014; French et al., 2011).
While GFED4 burned-area estimates come with regional
uncertainty estimates (Giglio et al., 2013), the actual uncertainty of C emissions from fires are probably much larger,
of the order of 50 % (van der Werf et al., 2017). The uncertainties of fire emission estimates depend regionally and
temporally on the various input data sets such as burned area,
small fire burned area, biomass loadings, and combustion
completeness. Better quantifying this uncertainty requires an
assessment of the burned-area estimates as well as new field
data on fuel consumption and emission factors. In this study
we cannot propagate this uncertainty into the NCE estimates
as this would require spatiotemporal error covariance matrices.
4.5
Seasonality for coastal and inland waters, wood
and crop harvest emissions
Recently, major steps have been undertaken to resolve the
spatial variability of coastal and inland water CO2 fluxes
(Laruelle et al., 2013, 2014; Lauerwald et al., 2015; Raymond et al., 2013). Estimates of the seasonal variation in
these fluxes are necessary to better constrain seasonal variations in NCE. For inland waters, seasonality has so far only
been investigated at regional scale (Laruelle et al., 2015;
Richey et al., 2002). For shelves some seasonal estimates
Biogeosciences, 14, 3685–3703, 2017
3698
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
are currently available in temperate and high latitudes, indicating that net C uptake is highest in spring, whereas C
release is highest in summer (Laruelle et al., 2014, 2017).
These estimates indicate that seasonal differences in shelf
net C exchange are as high as the annually integrated latitudinal gradient. An analysis performed over Atlantic shelves
suggests that the seasonal variability in the air–sea CO2 exchange is most pronounced over temperate latitudes. In these
regions, shelves generally behave as strong CO2 sinks in
winter and spring, partly sustained by CO2 fixation during
the spring phytoplankton bloom, but can become mild CO2
sources to the atmosphere in summer due to the effect of
temperature-driven decrease CO2 solubility in water (Laruelle et al., 2014). Such behaviour is consistent with that of
the open ocean at similar latitudes (Takahashi et al., 2009).
In the continental shelves surrounding other oceanic basins,
however, a recent study suggests more complex seasonal patterns involving the contributions of processes other than temperature to the seasonality of coastal pCO2 (Laruelle et al.,
2017).
Biogenic C emissions related to tropical aboveground
biomass loss as well as crop and wood harvest were equally
distributed across months in this study. When exactly C emissions from humans and livestock occur is difficult to predict
and would require more detailed consumption data (Wolf et
al., 2015a).
5
Conclusions
From the presented synthesis, we draw the following main
conclusions:
i. Current estimates of surface–atmosphere CO2 exchanges that are spatiotemporally explicit and entirely
driven by surface observation are not sufficiently well
constrained to close the C budget at the global scale.
The data-driven estimates show a large bias towards
too much C uptake by the Earth surface of nearly
10 PgC yr−1 .
ii. The most likely candidate for inducing the mismatch
between data-driven surface–atmosphere CO2 exchange
and the atmospheric CO2 growth rate is land NEP. In
particular, tropical NEP estimates appear to be strongly
overestimated (too large land sink). Understanding this
bias will help designing better upscaling approaches
(e.g. by including currently missing relevant predictors)
and pinpointing variables that need to be (better) monitored in the future.
iii. Regionally, the estimates of NCE are partly well constrained and may be used for model–data integration
studies, validation of models, and evaluating claims and
potentials of net C uptake within the framework of the
Paris Agreement (UNFCCC, 2015). These regions include Europe, Russia, South Asia, East Asia, Australia,
Biogeosciences, 14, 3685–3703, 2017
and most oceanic regions. Better constraining C fluxes
in regions with currently high uncertainties should be a
priority of future research.
Data availability. All variables used in Eq. (1) (including all ensembles runs listed in Table 1) are available for download (2001–
2010, monthly, 1◦ ). For NCE we provide the 200 ensemble runs
which are used in this study. The landing page of the DOI is a
data portal. After registration any user can download the data without restriction (https://dx.doi.org/10.17871/GEOCARBON_synth_
obs_v2).
The Supplement related to this article is available online
at https://doi.org/10.5194/bg-14-3685-2017-supplement.
Competing interests. The authors declare that they have no conflict
of interest.
Acknowledgements. This study was funded by the European
Union in the context of the FP7 project GEOCARBON (grant
agreement #283080). Authors affiliated with [2] and [16] further
acknowledge the EU for support via the H2020 project BACI
(grant agreement #640176). Authors affiliated with [5] thank
the SAMPLES project as part of the CGIAR research program
CCAFS, and CIFOR from the governments of Australia (grant
agreement #46167) and Norway (grant agreement #QZA-10/0468).
Nuno Carvalhais acknowledges funding from the NOVA grant
UID/AMB/04085/2013. Goulven G. Laruelle is “Chargé de
recherches F.R.S.-FNRS” at ULB. Ronny Lauerwald received
funding from ANR (ANR-10-LABX-0018) and BRIC at ULB.
Jens Hartmann and Ronny Lauerwald received funding from the
German Science Foundation (DFG) (HA4472/6-1 and the cluster of
excellence CLISAP, DFG EXEC 177, Hamburg). Leonardo Calle
acknowledges funding from a NASA Earth and Space Science
Fellowship. We also acknowledge the European Commission, Joint
Research Centre (JRC)/Netherlands Environmental Assessment
Agency (PBL) for providing the Emission Database for Global
Atmospheric Research (EDGAR), release version EDGARv4.2.
The article processing charges for this open-access
publication were covered by the Max Planck Society.
Edited by: Trevor Keenan
Reviewed by: two anonymous referees
References
Amiro, B. D., Barr, A. G., Barr, J. G., Black, T. A., Bracho, R.,
Brown, M., Chen, J., Clark, K. L., Davis, K. J., Desai, A. R.,
Dore, S., Engel, V., Fuentes, J. D., Goldstein, A. H., Goulden,
M. L., Kolb, T. E., Lavigne, M. B., Law, B. E., Margolis, H.
www.biogeosciences.net/14/3685/2017/
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
A., Martin, T., McCaughey, J. H., Misson, L., Montes-Helu,
M., Noormets, A., Randerson, J. T., Starr, G., and Xiao, J.:
Ecosystem carbon dioxide fluxes after disturbance in forests
of North America, J. Geophys. Res.-Biogeo., 115, G00K02,
https://doi.org/10.1029/2010JG001390, 2010.
Andreae, M. O., Acevedo, O. C., Araùjo, A., Artaxo, P., Barbosa, C.
G. G., Barbosa, H. M. J., Brito, J., Carbone, S., Chi, X., Cintra,
B. B. L., da Silva, N. F., Dias, N. L., Dias-Júnior, C. Q., Ditas, F.,
Ditz, R., Godoi, A. F. L., Godoi, R. H. M., Heimann, M., Hoffmann, T., Kesselmeier, J., Könemann, T., Krüger, M. L., Lavric,
J. V., Manzi, A. O., Lopes, A. P., Martins, D. L., Mikhailov, E.
F., Moran-Zuloaga, D., Nelson, B. W., Nölscher, A. C., Santos
Nogueira, D., Piedade, M. T. F., Pöhlker, C., Pöschl, U., Quesada, C. A., Rizzo, L. V., Ro, C. U., Ruckteschler, N., Sá, L. D.
A., de Oliveira Sá, M., Sales, C. B., dos Santos, R. M. N., Saturno, J., Schöngart, J., Sörgel, M., de Souza, C. M., de Souza,
R. A. F., Su, H., Targhetta, N., Tóta, J., Trebs, I., Trumbore,
S., van Eijck, A., Walter, D., Wang, Z., Weber, B., Williams,
J., Winderlich, J., Wittmann, F., Wolff, S., and Yáñez-Serrano,
A. M.: The Amazon Tall Tower Observatory (ATTO): overview
of pilot measurements on ecosystem ecology, meteorology, trace
gases, and aerosols, Atmos. Chem. Phys., 15, 10723–10776,
https://doi.org/10.5194/acp-15-10723-2015, 2015..
Atlas, R., Hoffman, R. N., Ardizzone, J., Leidner, S. M., Jusem, J.
C., Smith, D. K., and Gombos, D.: A Cross-Calibrated Multiplatform Ocean Surface Wind Velocity Product for Meteorological and Oceanographic Applications, B. Am. Meteorol. Soc., 92,
157–174, 2011.
Aufdenkampe, A. K., Mayorga, E., Raymond, P. A., Melack, J. M.,
Doney, S. C., Alin, S. R., Aalto, R. E., and Yoo, K.: Riverine
coupling of biogeochemical cycles between land, oceans and atmosphere, Front. Ecol. Environ., 9, 53–60, 2011.
Bakker, D. C. E., Pfeil, B., Smith, K., Hankin, S., Olsen, A., Alin,
S. R., Cosca, C., Harasawa, S., Kozyr, A., Nojiri, Y., O’Brien,
K. M., Schuster, U., Telszewski, M., Tilbrook, B., Wada, C.,
Akl, J., Barbero, L., Bates, N. R., Boutin, J., Bozec, Y., Cai,
W. J., Castle, R. D., Chavez, F. P., Chen, L., Chierici, M., Currie, K., de Baar, H. J. W., Evans, W., Feely, R. A., Fransson,
A., Gao, Z., Hales, B., Hardman-Mountford, N. J., Hoppema,
M., Huang, W. J., Hunt, C. W., Huss, B., Ichikawa, T., Johannessen, T., Jones, E. M., Jones, S. D., Jutterström, S., Kitidis,
V., Körtzinger, A., Landschützer, P., Lauvset, S. K., Lefèvre, N.,
Manke, A. B., Mathis, J. T., Merlivat, L., Metzl, N., Murata,
A., Newberger, T., Omar, A. M., Ono, T., Park, G. H., Paterson, K., Pierrot, D., Ríos, A. F., Sabine, C. L., Saito, S., Salisbury, J., Sarma, V. V. S. S., Schlitzer, R., Sieger, R., Skjelvan,
I., Steinhoff, T., Sullivan, K. F., Sun, H., Sutton, A. J., Suzuki,
T., Sweeney, C., Takahashi, T., Tjiputra, J., Tsurushima, N., van
Heuven, S. M. A. C., Vandemark, D., Vlahos, P., Wallace, D. W.
R., Wanninkhof, R., and Watson, A. J.: An update to the Surface
Ocean CO2 Atlas (SOCAT version 2), Earth Syst. Sci. Data, 6,
69–90, https://doi.org/10.5194/essd-6-69-2014, 2014.
Baldocchi, D.: Measuring fluxes of trace gases and energy between
ecosystems and the atmosphere – the state and future of the eddy
covariance method, Glob. Change Biol., 20, 3600–3609, 2014.
Ballantyne, A. P., Alden, C. B., Miller, J. B., Tans, P. P., and White,
J. W. C.: Increase in observed net carbon dioxide uptake by land
and oceans during the past 50 years, Nature, 488, 70–72, 2012.
www.biogeosciences.net/14/3685/2017/
3699
Berger, M., Moreno, J., Johannessen, J. A., Levelt, P. F., and
Hanssen, R. F.: ESA’s sentinel missions in support of Earth system science, Remote Sens. Environ., 120, 84–90, 2012.
Canadell, J. G., Ciais, P., Gurney, K., Le Quéré, C., Piao, S., Raupach, M. R., and Sabine, C. L.: An International Effort to Quantify Regional Carbon Fluxes, EOS, 92, 81–82, 2011.
Ciais, P., Dolman, A., Bombelli, A., Duren, R., Peregon, A.,
Rayner, P., Miller, C., Gobron, N., Kinderman, G., and Marland, G.: Current systematic carbon-cycle observations and the
need for implementing a policy-relevant carbon observing system, Biogeosciences, 11, 3547–3602, https://doi.org/10.5194/bg11-3547-2014, 2014.
Ciais, P., Gasser, T., Lauerwald, R., Peng, S., Raymond, P. A.,
Wang, Y., Canadell, J. G., Peters, G. P., Andres, R. J., Chang,
J., Yue, C., Dolman, A. J., Haverd, V., Hartmann, J., Laruelle,
G., King, A. W., Liu, Y., Luyssaert, S., Maignan, F., Patra, P.
K., Peregon, A., Regnier, P., Piao, S., Poulter, B., Shvidenko, A.,
Valentini, R., Wang, R., van Dijk, A. I. J. M., Broquet, G., Yin, Y.,
Zscheischler, J., and Zhu, D.: Observed regional carbon budgets
imply reduced soil heterotrophic respiration, Nature, in review,
2017.
Ciais, P., Sabine, C., Bala, G., Bopp, L., Brovkin, V., Canadell, J.,
Chhabra, A., DeFries, R., Galloway, J., Heimann, M., Jones, C.,
Le Queìreì, C., Myneni, R. B., Piao, S., and Thornton, P.: Carbon
and Other Biogeochemical Cycles, in: Climate Change 2013:
The Physical Science Basis, Contribution of Working Group I to
the Fifth Assessment Report of the Intergovernmental Panel on
Climate Change, edited by: Stocker, T. F., Qin, D., Plattner, G.K., Tignor, M., Allen, S. K., Boschung, J., Nauels, A., Xia, Y.,
Bex, V., and Midgley, P. M., Cambridge University Press, Cambridge, United Kingdom and New York, NY, USA, 2013.
Coursolle, C., Margolis, H. A., Giasson, M. A., Bernier, P. Y.,
Amiro, B. D., Arain, M. A., Barr, A. G., Black, T. A., Goulden,
M. L., McCaughey, J. H., Chen, J. M., Dunn, A. L., Grant, R.
F., and Lafleur, P. M.: Influence of stand age on the magnitude
and seasonality of carbon fluxes in Canadian forests, Agr. Forest
Meteorol., 165, 136–148, 2012.
Dee, D. P., Uppala, S. M., Simmons, A. J., Berrisford, P., Poli,
P., Kobayashi, S., Andrae, U., Balmaseda, M. A., Balsamo, G.,
Bauer, P., Bechtold, P., Beljaars, A. C. M., van de Berg, L., Bidlot, J., Bormann, N., Delsol, C., Dragani, R., Fuentes, M., Geer,
A. J., Haimberger, L., Healy, S. B., Hersbach, H., Holm, E. V.,
Isaksen, L., Kallberg, P., Kohler, M., Matricardi, M., McNally,
A. P., Monge-Sanz, B. M., Morcrette, J. J., Park, B. K., Peubey,
C., de Rosnay, P., Tavolato, C., Thepaut, J. N., and Vitart, F.: The
ERA-Interim reanalysis: configuration and performance of the
data assimilation system, Q. J. Roy. Meteor. Soc., 137, 553–597,
2011.
Dolman, A. J., Shvidenko, A., Schepaschenko, D., Ciais, P.,
Tchebakova, N., Chen, T., van der Molen, M. K., Belelli Marchesini, L., Maximov, T. C., Maksyutov, S., and Schulze, E. D.:
An estimate of the terrestrial carbon budget of Russia using
inventory-based, eddy covariance and inversion methods, Biogeosciences, 9, 5323–5340, https://doi.org/10.5194/bg-9-53232012, 2012.
Dürr, H. H., Laruelle, G. G., van Kempen, C. M., Slomp, C.
P., Meybeck, M., and Middelkoop, H.: Worldwide Typology
of Nearshore Coastal Systems: Defining the Estuarine Filter of
River Inputs to the Oceans, Estuar. Coast., 34, 441–458, 2011.
Biogeosciences, 14, 3685–3703, 2017
3700
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
Earles, J. M., Yeh, S., and Skog, K. E.: Timing of carbon emissions
from global forest clearance, Nature Climate Change, 2, 682–
685, 2012.
Fekete, B. M., Vorosmarty, C. J., and Grabs, W.: High-resolution
fields of global runoff combining observed river discharge
and simulated water balances, Global Biogeochem. Cy., 16,
https://doi.org/10.1029/1999GB001254, 2002.
French, N. H. F., de Groot, W. J., Jenkins, L. K., Rogers, B. M., Alvarado, E., Amiro, B., de Jong, B., Goetz, S., Hoy, E., Hyer, E.,
Keane, R., Law, B. E., McKenzie, D., McNulty, S. G., Ottmar,
R., Perez-Salicrup, D. R., Randerson, J., Robertson, K. M., and
Turetsky, M.: Model comparisons for estimating carbon emissions from North American wildland fire, J. Geophys. Res.Biogeo., 116, G00K05, https://doi.org/10.1029/2010JG001469,
2011.
Giglio, L., Randerson, J. T., and van der Werf, G. R.: Analysis of daily, monthly, and annual burned area using the fourthgeneration global fire emissions database (GFED4), J. Geophys.
Res.-Biogeo., 118, 317–328, 2013.
GLOBALVIEW-CO2: Cooperative atmospheric data integration
project-Carbon dioxide. Multi-laboratory compilation of synchronized and gap-filled atmospheric carbon dioxide records for
the period 1979–2011, NOAA ESRL, Boulder, 2012.
Gloor, M., Gatti, L., Brienen, R., Feldpausch, T. R., Phillips, O.
L., Miller, J., Ometto, J. P., Rocha, H., Baker, T., de Jong, B.,
Houghton, R. A., Malhi, Y., Aragão, L. E. O. C., Guyot, J. L.,
Zhao, K., Jackson, R., Peylin, P., Sitch, S., Poulter, B., Lomas,
M., Zaehle, S., Huntingford, C., Levy, P., and Lloyd, J.: The carbon balance of South America: a review of the status, decadal
trends and main determinants, Biogeosciences, 9, 5407–5430,
https://doi.org/10.5194/bg-9-5407-2012, 2012.
Harris, N. L., Brown, S., Hagen, S. C., Saatchi, S. S., Petrova, S.,
Salas, W., Hansen, M. C., Potapov, P. V., and Lotsch, A.: Baseline Map of Carbon Emissions from Deforestation in Tropical
Regions, Science, 336, 1573–1576, 2012.
Hartmann, J., Lauerwald, R., and Moosdorf, N.: A brief overview
of the GLObal RIver CHemistry Database, GLORICH, Procedia
Earth and Planetary Science, 10, 23–27, 2014.
Haverd, V., Raupach, M. R., Briggs, P. R., Canadell, J. G., Davis, S.
J., Law, R. M., Meyer, C. P., Peters, G. P., Pickett-Heaps, C., and
Sherman, B.: The Australian terrestrial carbon budget, Biogeosciences, 10, 851–869, https://doi.org/10.5194/bg-10-851-2013,
2013.
Hayes, D. J., Turner, D. P., Stinson, G., McGuire, A. D., Wei, Y.,
West, T. O., Heath, L. S., de Jong, B., McConkey, B. G., Birdsey,
R. A., Kurz, W. A., Jacobson, A. R., Huntzinger, D. N., Pan, Y.,
Post, W. M., and Cook, R. B.: Reconciling estimates of the contemporary North American carbon balance among terrestrial biosphere models, atmospheric inversions, and a new approach for
estimating net ecosystem exchange from inventory-based data,
Glob. Change Biol., 18, 1282–1299, 2012.
Houghton, R. A.: Balancing the global carbon budget, Annu. Rev.
Earth Pl. Sc., 35, 313–347, 2007.
Houghton, R. A. and Nassikas, A. A.: Global and regional fluxes of
carbon from land use and land cover change 1850–2015, Global
Biogeochem. Cy., 31, 456–472, 2017.
Hyvönen, R., Agren, G. I., Linder, S., Persson, T., Cotrufo, M. F.,
Ekblad, A., Freeman, M., Grelle, A., Janssens, I. A., Jarvis, P. G.,
Kellomaki, S., Lindroth, A., Loustau, D., Lundmark, T., Norby,
Biogeosciences, 14, 3685–3703, 2017
R. J., Oren, R., Pilegaard, K., Ryan, M. G., Sigurdsson, B. D.,
Stromgren, M., van Oijen, M., and Wallin, G.: The likely impact of elevated [CO2 ], nitrogen deposition, increased temperature and management on carbon sequestration in temperate and
boreal forest ecosystems: a literature review, New Phytol., 173,
463–480, 2007.
Ishii, M., Feely, R. A., Rodgers, K. B., Park, G.-H., Wanninkhof, R.,
Sasano, D., Sugimoto, H., Cosca, C. E., Nakaoka, S., Telszewski,
M., Nojiri, Y., Mikaloff Fletcher, S. E., Niwa, Y., Patra, P. K.,
Valsala, V., Nakano, H., Lima, I., Doney, S. C., Buitenhuis, E. T.,
Aumont, O., Dunne, J. P., Lenton, A., and Takahashi, T.: Air–sea
CO2 flux in the Pacific Ocean for the period 1990–2009, Biogeosciences, 11, 709–734, https://doi.org/10.5194/bg-11-709-2014,
2014.
Jägermeyr, J., Gerten, D., Lucht, W., Hostert, P., Migliavacca,
M., and Nemani, R.: A high-resolution approach to estimating ecosystem respiration at continental scales using operational
satellite data, Glob. Change Biol., 20, 1191–1210, 2014.
Jähne, B., Munnich, K. O., Bosinger, R., Dutzi, A., Huber, W.,
and Libner, P.: On the Parameters Influencing Air-Water GasExchange, J. Geophys. Res.-Ocean., 92, 1937–1949, 1987.
Jung, M., Reichstein, M., Margolis, H. A., Cescatti, A., Richardson, A. D., Arain, M. A., Arneth, A., Bernhofer, C., Bonal,
D., Chen, J. Q., Gianelle, D., Gobron, N., Kiely, G., Kutsch,
W., Lasslop, G., Law, B. E., Lindroth, A., Merbold, L.,
Montagnani, L., Moors, E. J., Papale, D., Sottocornola, M.,
Vaccari, F., and Williams, C.: Global patterns of landatmosphere fluxes of carbon dioxide, latent heat, and sensible heat derived from eddy covariance, satellite, and meteorological observations, J. Geophys. Res.-Biogeo., 116, G00J07,
https://doi.org/10.1029/2010JG001566, 2011.
King, A. W., Andres, R. J., Davis, K. J., Hafer, M., Hayes, D. J.,
Huntzinger, D. N., de Jong, B., Kurz, W. A., McGuire, A. D., Vargas, R., Wei, Y., West, T. O., and Woodall, C. W.: North America’s net terrestrial CO2 exchange with the atmosphere 1990–
2009, Biogeosciences, 12, 399–414, https://doi.org/10.5194/bg12-399-2015, 2015..
Kondo, M., Ichii, K., Takagi, H., and Sasakawa, M.: Comparison of
the data-driven top-down and bottom-up global terrestrial CO2
exchanges: GOSAT CO2 inversion and empirical eddy flux upscaling, J. Geophys. Res.-Biogeo., 120, 1226–1245, 2015.
Landschützer, P., Gruber, N., Bakker, D. C. E., and Schuster, U.:
Recent variability of the global ocean carbon sink, Global Biogeochem. Cy., 28, 927–949, 2014.
Landschützer, P., Gruber, N., Haumann, F. A., Rödenbeck, C.,
Bakker, D. C. E., van Heuven, S., Hoppema, M., Metzl, N.,
Sweeney, C., Takahashi, T., Tilbrook, B., and Wanninkhof, R.:
The reinvigoration of the Southern Ocean carbon sink, Science,
349, 1221–1224, 2015.
Laruelle, G. G., Durr, H. H., Lauerwald, R., Hartmann, J., Slomp,
C. P., Goossens, N., and Regnier, P. A. G.: Global multi-scale
segmentation of continental and coastal waters from the watersheds to the continental margins, Hydrol. Earth Syst. Sci., 17,
2029–2051, https://doi.org/10.5194/hess-17-2029-2013, 2013.
Laruelle, G. G., Lauerwald, R., Pfeil, B., and Regnier, P.: Regionalized global budget of the CO2 exchange at the air-water interface in continental shelf seas, Global Biogeochem. Cy., 28,
1199–1214, 2014.
www.biogeosciences.net/14/3685/2017/
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
Laruelle, G. G., Lauerwald, R., Rotschi, J., Raymond, P. A., Hartmann, J., and Regnier, P.: Seasonal response of air-water CO2
exchange along the land-ocean aquatic continuum of the northeast North American coast, Biogeosciences, 12, 1447–1458,
https://doi.org/10.5194/bg-12-1447-2015, 2015.
Laruelle, G. G., Landschützer, P., Gruber, N., Tison, J.-L., Delille,
B., and Regnier, P.: Global high resolution monthly pCO2 climatology for the coastal ocean derived from neural network interpolation, Biogeosciences Discuss., https://doi.org/10.5194/bg2017-64, in review, 2017.
Lauerwald, R., Laruelle, G. G., Hartmann, J., Ciais, P., and Regnier,
P. A. G.: Spatial patterns in CO2 evasion from the global river
network, Global Biogeochem. Cy., 29, 534–554, 2015.
Le Quéré, C., Moriarty, R., Andrew, R. M., Canadell, J. G., Sitch, S.,
Korsbakken, J. I., Friedlingstein, P., Peters, G. P., Andres, R. J.,
Boden, T. A., Houghton, R. A., House, J. I., Keeling, R. F., Tans,
P., Arneth, A., Bakker, D. C. E., Barbero, L., Bopp, L., Chang,
J., Chevallier, F., Chini, L. P., Ciais, P., Fader, M., Feely, R. A.,
Gkritzalis, T., Harris, I., Hauck, J., Ilyina, T., Jain, A. K., Kato,
E., Kitidis, V., Klein Goldewijk, K., Koven, C., Landschützer,
P., Lauvset, S. K., Lefèvre, N., Lenton, A., Lima, I. D., Metzl,
N., Millero, F., Munro, D. R., Murata, A., Nabel, J. E. M. S.,
Nakaoka, S., Nojiri, Y., O’Brien, K., Olsen, A., Ono, T., Pérez,
F. F., Pfeil, B., Pierrot, D., Poulter, B., Rehder, G., Rödenbeck,
C., Saito, S., Schuster, U., Schwinger, J., Séférian, R., Steinhoff,
T., Stocker, B. D., Sutton, A. J., Takahashi, T., Tilbrook, B., van
der Laan-Luijkx, I. T., van der Werf, G. R., van Heuven, S., Vandemark, D., Viovy, N., Wiltshire, A., Zaehle, S., and Zeng, N.:
Global Carbon Budget 2015, Earth Syst. Sci. Data, 7, 349–396,
https://doi.org/10.5194/essd-7-349-2015, 2015.
Lehner, B. and Döll, P.: Development and validation of a global
database of lakes, reservoirs and wetlands, J. Hydrol., 296, 1–22,
2004.
Lehner, B., Verdin, K., and Jarvis, A.: New Global Hydrography Derived From Spaceborne Elevation Data, EOS, 89, 93–94,
2008.
Lenton, A., Tilbrook, B., Law, R. M., Bakker, D., Doney, S. C.,
Gruber, N., Ishii, M., Hoppema, M., Lovenduski, N. S., Matear,
R. J., McNeil, B. I., Metzl, N., Mikaloff Fletcher, S. E., Monteiro,
P. M. S., Rödenbeck, C., Sweeney, C., and Takahashi, T.: Sea–
air CO2 fluxes in the Southern Ocean for the period 1990–2009,
Biogeosciences, 10, 4037–4054, https://doi.org/10.5194/bg-104037-2013, 2013.
Luyssaert, S., Abril, G., Andres, R., Bastviken, D., Bellassen, V.,
Bergamaschi, P., Bousquet, P., Chevallier, F., Ciais, P., Corazza,
M., Dechow, R., Erb, K. H., Etiope, G., Fortems-Cheiney, A.,
Grassi, G., Hartmann, J., Jung, M., Lathière, J., Lohila, A., Mayorga, E., Moosdorf, N., Njakou, D. S., Otto, J., Papale, D., Peters,
W., Peylin, P., Raymond, P., Rödenbeck, C., Saarnio, S., Schulze,
E. D., Szopa, S., Thompson, R., Verkerk, P. J., Vuichard, N.,
Wang, R., Wattenbach, M., and Zaehle, S.: The European land
and inland water CO2 , CO, CH4 and N2 O balance between 2001
and 2005, Biogeosciences, 9, 3357–3380, 2012.
Magnani, F., Mencuccini, M., Borghetti, M., Berbigier, P.,
Berninger, F., Delzon, S., Grelle, A., Hari, P., Jarvis, P. G., Kolari, P., Kowalski, A. S., Lankreijer, H., Law, B. E., Lindroth, A.,
Loustau, D., Manca, G., Moncrieff, J. B., Rayment, M., Tedeschi,
V., Valentini, R., and Grace, J.: The human footprint in the car-
www.biogeosciences.net/14/3685/2017/
3701
bon cycle of temperate and boreal forests, Nature, 447, 848–850,
2007.
Malhi, Y., Phillips, O. L., Lloyd, J., Baker, T., Wright, J., Almeida,
S., Arroyo, L., Frederiksen, T., Grace, J., Higuchi, N., Killeen,
T., Laurance, W. F., Leano, C., Lewis, S., Meir, P., Monteagudo,
A., Neill, D., Vargas, P. N., Panfil, S. N., Patino, S., Pitman, N.,
Quesada, C. A., Rudas-Ll, A., Salomao, R., Saleska, S., Silva,
N., Silveira, M., Sombroek, W. G., Valencia, R., Martinez, R.
V., Vieira, I. C. G., and Vinceti, B.: An international network to
monitor the structure, composition and dynamics of Amazonian
forests (RAINFOR), J. Veg. Sci., 13, 439–450, 2002.
Masarie, K. A. and Tans, P. P.: Extension and Integration of Atmospheric Carbon-Dioxide Data into a Globally Consistent Measurement Record, J. Geophys. Res.-Atmos., 100, 11593–11610,
1995.
Meybeck, M., Durr, H. H., and Vorosmarty, C. J.: Global coastal
segmentation and its river catchment contributors: A new look
at land-ocean linkage, Global Biogeochem. Cy., 20, GB1S90,
https://doi.org/10.1029/2005GB002540, 2006.
Pan, Y., Birdsey, R. A., Fang, J., Houghton, R., Kauppi, P. E., Kurz,
W. A., Phillips, O. L., Shvidenko, A., Lewis, S. L., Canadell,
J. G., Ciais, P., Jackson, R. B., Pacala, S. W., McGuire, A. D.,
Piao, S., Rautiainen, A., Sitch, S., and Hayes, D.: A Large and
Persistent Carbon Sink in the World’s Forests, Science, 333, 988–
993, 2011.
Patra, P. K., Canadell, J. G., Houghton, R. A., Piao, S. L., Oh, N.
H., Ciais, P., Manjunath, K. R., Chhabra, A., Wang, T., Bhattacharya, T., Bousquet, P., Hartman, J., Ito, A., Mayorga, E.,
Niwa, Y., Raymond, P. A., Sarma, V. V. S. S., and Lasco, R.:
The carbon budget of South Asia, Biogeosciences, 10, 513–527,
https://doi.org/10.5194/bg-10-513-2013, 2013.
Peylin, P., Law, R. M., Gurney, K. R., Chevallier, F., Jacobson,
A. R., Maki, T., Niwa, Y., Patra, P. K., Peters, W., Rayner, P.
J., Rodenbeck, C., van der Laan-Luijkx, I. T., and Zhang, X.:
Global atmospheric carbon budget: results from an ensemble of
atmospheric CO2 inversions, Biogeosciences, 10, 6699–6720,
https://doi.org/10.5194/bg-10-6699-2013, 2013.
Piao, S. L., Ito, A., Li, S. G., Huang, Y., Ciais, P., Wang, X. H.,
Peng, S. S., Nan, H. J., Zhao, C., Ahlström, A., Andres, R. J.,
Chevallier, F., Fang, J. Y., Hartmann, J., Huntingford, C., Jeong,
S., Levis, S., Levy, P. E., Li, J. S., Lomas, M. R., Mao, J. F.,
Mayorga, E., Mohammat, A., Muraoka, H., Peng, C. H., Peylin,
P., Poulter, B., Shen, Z. H., Shi, X., Sitch, S., Tao, S., Tian, H.
Q., Wu, X. P., Xu, M., Yu, G. R., Viovy, N., Zaehle, S., Zeng, N.,
and Zhu, B.: The carbon budget of terrestrial ecosystems in East
Asia over the last two decades, Biogeosciences, 9, 3571–3586,
https://doi.org/10.5194/bg-9-3571-2012, 2012.
Pongratz, J., Reick, C. H., Raddatz, T., and Claussen, M.: Effects of anthropogenic land cover change on the carbon cycle
of the last millennium, Global Biogeochem. Cy., 23, GB4001,
https://doi.org/10.1029/2009GB003488, 2009.
Poulter, B.: Global Wood Harvest, Operational Global Carbon Observing System – GEOCARBON Project Report, 2015 (data
available upon request).
Prather, M. J., Holmes, C. D., and Hsu, J.: Reactive greenhouse
gas scenarios: Systematic exploration of uncertainties and the
role of atmospheric chemistry, Geophys. Res. Lett., 39, L09803,
https://doi.org/10.1029/2012GL051440, 2012.
Biogeosciences, 14, 3685–3703, 2017
3702
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
Randerson, J. T., Chen, Y., van der Werf, G. R., Rogers, B. M.,
and Morton, D. C.: Global burned area and biomass burning
emissions from small fires, J. Geophys. Res., 117, G04012,
https://doi.org/10.1029/2012JG002128, 2012.
Raupach, M. R., Rayner, P. J., Barrett, D. J., DeFries, R. S.,
Heimann, M., Ojima, D. S., Quegan, S., and Schmullius, C.
C.: Model-data synthesis in terrestrial carbon observation: methods, data requirements and data uncertainty specifications, Glob.
Change Biol., 11, 378–397, 2005.
Raymond, P. A., Hartmann, J., Lauerwald, R., Sobek, S., McDonald, C., Hoover, M., Butman, D., Striegl, R., Mayorga, E., Humborg, C., Kortelainen, P., Durr, H., Meybeck, M., Ciais, P., and
Guth, P.: Global carbon dioxide emissions from inland waters,
Nature, 503, 355–359, 2013.
Raymond, P. A., Zappa, C. J., Butman, D., Bott, T. L., Potter, J.,
Mulholland, P., Laursen, A. E., McDowell, W. H., and Newbold,
D.: Scaling the gas transfer velocity and hydraulic geometry in
streams and small rivers, Limnol. Oceanogr., 2, 41–53, 2012.
Rayner, P. J., Scholze, M., Knorr, W., Kaminski, T., Giering, R., and Widmann, H.: Two decades of terrestrial
carbon fluxes from a carbon cycle data assimilation system (CCDAS), Global Biogeochem. Cy., 19, GB2026,
https://doi.org/10.1029/2004GB002254, 2005.
Richey, J. E., Melack, J. M., Aufdenkampe, A. K., Ballester, V. M.,
and Hess, L. L.: Outgassing from Amazonian rivers and wetlands
as a large tropical source of atmospheric CO2 , Nature, 416, 617–
620, 2002.
Rödenbeck, C., Bakker, D. C. E., Gruber, N., Iida, Y., Jacobson, A. R., Jones, S., Landschützer, P., Metzl, N., Nakaoka,
S., Olsen, A., Park, G. H., Peylin, P., Rodgers, K. B., Sasse,
T. P., Schuster, U., Shutler, J. D., Valsala, V., Wanninkhof, R.,
and Zeng, J.: Data-based estimates of the ocean carbon sink
variability first – results of the Surface Ocean pCO2 Mapping intercomparison (SOCOM), Biogeosciences, 12, 7251–
7278, https://doi.org/10.5194/bg-12-7251-2015, 2015..
Rödenbeck, C., Bakker, D. C. E., Metzl, N., Olsen, A., Sabine,
C., Cassar, N., Reum, F., Keeling, R. F., and Heimann, M.:
Interannual sea-air CO2 flux variability from an observationdriven ocean mixed-layer scheme, Biogeosciences, 11, 4599–
4613, https://doi.org/10.5194/bg-11-4599-2014, 2014.
Ruddiman, W. F.: The anthropogenic greenhouse era began thousands of years ago, Climatic Change, 61, 261–293, 2003.
Sarma, V. V. S. S., Lenton, A., Law, R. M., Metzl, N., Patra, P. K., Doney, S., Lima, I. D., Dlugokencky, E., Ramonet, M., and Valsala, V.: Sea–air CO2 fluxes in the Indian
Ocean between 1990 and 2009, Biogeosciences, 10, 7035–7052,
https://doi.org/10.5194/bg-10-7035-2013, 2013.
Schuster, U., McKinley, G. A., Bates, N., Chevallier, F., Doney,
S. C., Fay, A. R., González-Dávila, M., Gruber, N., Jones,
S., Krijnen, J., Landschützer, P., Lefèvre, N., Manizza, M.,
Mathis, J., Metzl, N., Olsen, A., Rios, A. F., Rödenbeck,
C., Santana-Casiano, J. M., Takahashi, T., Wanninkhof, R.,
and Watson, A. J.: An assessment of the Atlantic and Arctic
sea–air CO2 fluxes, 1990–2009, Biogeosciences, 10, 607–627,
https://doi.org/10.5194/bg-10-607-2013, 2013.
Schwalm, C. R., Huntzinger, D. N., Fisher, J. B., Michalak, A. M.,
Bowman, K., Ciais, P., Cook, R., El-Masri, B., Hayes, D., Huang,
M., Ito, A., Jain, A., King, A. W., Lei, H., Liu, J., Lu, C., Mao, J.,
Peng, S., Poulter, B., Ricciuto, D., Schaefer, K., Shi, X., Tao, B.,
Biogeosciences, 14, 3685–3703, 2017
Tian, H., Wang, W., Wei, Y., Yang, J., and Zeng, N.: Toward “optimal” integration of terrestrial biosphere models, Geophys. Res.
Lett., 42, 4418–4428, https://doi.org/10.1002/2015GL064002,
2015.
Sindelarova, K., Granier, C., Bouarar, I., Guenther, A., Tilmes, S.,
Stavrakou, T., Muller, J. F., Kuhn, U., Stefani, P., and Knorr, W.:
Global data set of biogenic VOC emissions calculated by the
MEGAN model over the last 30 years, Atmos. Chem. Phys., 14,
9317–9341, https://doi.org/10.5194/acp-14-9317-2014, 2014.
Sitch, S., Friedlingstein, P., Gruber, N., Jones, S. D., MurrayTortarolo, G., Ahlström, A., Doney, S. C., Graven, H., Heinze,
C., Huntingford, C., Levis, S., Levy, P. E., Lomas, M., Poulter, B., Viovy, N., Zaehle, S., Zeng, N., Arneth, A., Bonan,
G., Bopp, L., Canadell, J. G., Chevallier, F., Ciais, P., Ellis,
R., Gloor, M., Peylin, P., Piao, S. L., Le Quéré, C., Smith, B.,
Zhu, Z., and Myneni, R.: Recent trends and drivers of regional
sources and sinks of carbon dioxide, Biogeosciences, 12, 653–
679, https://doi.org/10.5194/bg-12-653-2015, 2015.
Takahashi, T., Sutherland, S. C., Wanninkhof, R., Sweeney, C.,
Feely, R. A., Chipman, D. W., Hales, B., Friederich, G., Chavez,
F., Sabine, C., Watson, A., Bakker, D. C. E., Schuster, U., Metzl,
N., Yoshikawa-Inoue, H., Ishii, M., Midorikawa, T., Nojiri, Y.,
Kortzinger, A., Steinhoff, T., Hoppema, M., Olafsson, J., Arnarson, T. S., Tilbrook, B., Johannessen, T., Olsen, A., Bellerby, R.,
Wong, C. S., Delille, B., Bates, N. R., and de Baar, H. J. W.: Climatological mean and decadal change in surface ocean pCO2 ,
and net sea-air CO2 flux over the global oceans, Deep-Sea Res.
Pt. I, 56, 2075–2076, 2009.
Tatem, A. J., Goetz, S. J., and Hay, S. I.: Fifty years of earthobservation satellites – Views from space have led to countless
advances on the ground in both scientific knowledge and daily
life, Am. Sci., 96, 390–398, 2008.
Tramontana, G., Jung, M., Schwalm, C. R., Ichii, K., Camps-Valls,
G., Ráduly, B., Reichstein, M., Arain, M. A., Cescatti, A., Kiely,
G., Merbold, L., Serrano-Ortiz, P., Sickert, S., Wolf, S., and
Papale, D.: Predicting carbon dioxide and energy fluxes across
global FLUXNET sites with regression algorithms, Biogeosciences, 13, 4291–4313, https://doi.org/10.5194/bg-13-42912016, 2016.
Trumbore, S.: Carbon respired by terrestrial ecosystems –recent
progress and challenges, Glob. Change Biol., 12, 141–153, 2006.
Tyukavina, A., Baccini, A., Hansen, M. C., Potapov, P. V., Stehman,
S. V., Houghton, R. A., Krylov, A. M., Turubanova, S., and
Goetz, S. J.: Aboveground carbon loss in natural and managed
tropical forests from 2000 to 2012, Environ. Res. Lett., 10,
074002, https://doi.org/10.1088/1748-9326/10/7/074002, 2015.
UNFCCC:
http://www4.unfccc.int/Submissions/INDC/
SubmissionPages/submissions.aspx (last accessed: 25 July
2017), 2015.
Valentini, R., Arneth, A., Bombelli, A., Castaldi, S., Cazzolla Gatti,
R., Chevallier, F., Ciais, P., Grieco, E., Hartmann, J., Henry,
M., Houghton, R. A., Jung, M., Kutsch, W. L., Malhi, Y., Mayorga, E., Merbold, L., Murray-Tortarolo, G., Papale, D., Peylin,
P., Poulter, B., Raymond, P. A., Santini, M., Sitch, S., Vaglio
Laurin, G., van der Werf, G. R., Williams, C. A., and Scholes,
R. J.: A full greenhouse gases budget of Africa: synthesis, uncertainties, and vulnerabilities, Biogeosciences, 11, 381–407,
https://doi.org/10.5194/bg-11-381-2014, 2014.
www.biogeosciences.net/14/3685/2017/
J. Zscheischler et al.: Limits of globally data-driven CO2 fluxes between surface and atmosphere
van der Werf, G. R., Randerson, J. T., Giglio, L., Collatz, G.
J., Mu, M., Kasibhatla, P. S., Morton, D. C., DeFries, R. S.,
Jin, Y., and van Leeuwen, T. T.: Global fire emissions and the
contribution of deforestation, savanna, forest, agricultural, and
peat fires (1997–2009), Atmos. Chem. Phys., 10, 11707–11735,
https://doi.org/10.5194/acp-10-11707-2010, 2010.
van der Werf, G. R., Randerson, J. T., Giglio, L., van Leeuwen, T.
T., Chen, Y., Rogers, B. M., Mu, M., van Marle, M. J. E., Morton,
D. C., Collatz, G. J., Yokelson, R. J., and Kasibhatla, P. S.: Global
fire emissions estimates during 1997–2015, Earth Syst. Sci. Data
Discuss., https://doi.org/10.5194/essd-2016-62, accepted, 2017.
Vernadsky, V.: The Biosphere (New York: Copernicus, Springer),
1926.
Wanninkhof, R.: Relationship between wind speed and gas exchange over the ocean, J. Geophys. Res.-Ocean., 97, 7373–7382,
1992.
Wanninkhof, R., Park, G. H., Takahashi, T., Sweeney, C., Feely,
R., Nojiri, Y., Gruber, N., Doney, S. C., McKinley, G. A.,
Lenton, A., Le Quéré, C., Heinze, C., Schwinger, J., Graven,
H., and Khatiwala, S.: Global ocean carbon uptake: magnitude, variability and trends, Biogeosciences, 10, 1983–2000,
https://doi.org/10.5194/bg-10-1983-2013, 2013..
www.biogeosciences.net/14/3685/2017/
3703
West, T. O., Bandaru, V., Brandt, C. C., Schuh, A. E., and
Ogle, S. M.: Regional uptake and release of crop carbon in the United States, Biogeosciences, 8, 2037–2046,
https://doi.org/10.5194/bg-8-2037-2011, 2011.
Wolf, J., West, T. O., Le Page, Y. L., Kyle, G. P., Zhang, X., Collatz,
G. J., and Imhoff, M. L.: Biogenic carbon fluxes from global agricultural production and consumption, Global Biogeochem. Cy.,
https://doi.org/10.1002/2015GB005119, 2015a.
Wolf, J., West, T. O., Le Page, Y. L., Kyle, G. P., Zhang, X., Collatz, G. J., and Imhoff, M. L.: CMS: Carbon Fluxes from Global
Agricultural Production and Consumption, 2005–2011, ORNL
DAAC, Oak Ridge, Tennessee, USA, 2015b.
Zhou, L. M., Tian, Y. H., Myneni, R. B., Ciais, P., Saatchi, S., Liu,
Y. Y., Piao, S. L., Chen, H. S., Vermote, E. F., Song, C. H., and
Hwang, T. H.: Widespread decline of Congo rainforest greenness
in the past decade, Nature, 509, 86–90, 2014.
Biogeosciences, 14, 3685–3703, 2017
|
|
https://openalex.org/W2110261674
|
https://zenodo.org/records/1275759/files/article.pdf
|
English
| null |
Live Training Transformation (LT2) product line applied standards for reusable integrated and interoperable solutions
| null | 2,008
|
public-domain
| 8,331
|
ABSTRACT The U.S. Army Program Executive Office (PEO) Simulation Training and Instrumentation (STRI) has established a Live
Training Transformation (LT2) product line approach to developing a Family of Training Systems (FTS) that provide the ground
maneuver training range functions supporting Army live and Joint training environments. The success of the LT2 program
strategy is dependent on defining standards and initiatives derived to promote systematic reuse of software and interoperability
solutions for the LT2 products. Application of LT2 product line standards have matured reducing life cycle development and
sustainment costs through the creation of common reusable architectures providing integrated and interoperable training
solutions for the LT2-FTS deployable to Army Ranges. LT2 standard initiatives cover the following primary areas of technology
development: Tactical Engagement Simulation System (TESS), training instrumentation systems, software architecture, targetry
and battlefield effects, and Joint Live-Virtual-Constructive interoperability. Approved and established standards include
Common Training Instrumentation Architecture (CTIA), OneTESS, Family of Army Systems and Integrated Targetry (FASIT),
Integrated Player Unit (IPU), Common Player Unit Interface Control Document (ICD), Consolidated Product Line Management
(CPM), Graphical User Interface (GUI) Framework, and LT2 HCI Style guide. The LT2 standards provide industry the
appropriate development guidelines and interface definitions in order to maximize industry involvement in developing product
line solutions and providing advanced training capabilities through technology insertion. This paper provides an overview of how
government and industry worked together to establish the LT2 standard initiatives based on common LT2-FTS use cases and
details the relevant solution sets resulting in achievement of the PEO STRI mission and the LT2-FTS product line
interoperability and reuse objectives. Live Training Transformation (LT2) Product Line Applied Standards For
Reusable Integrated And Interoperable Solutions Barry Clinger
Riptide Software, Inc. Orlando, Florida Jorge Rivera, William Samper
U.S. Army PEO STRI
Orlando, Florida
Jorge.Rivera2@peostri.army.mil,
william.samper@peostri.army.mil Barry.Clinger@Riptidesoftware.com MILCOM 2008 MILCOM 2008 U.S. Government work not protected by U.S. copyright. Live Training Domain Problem Space Live training range systems provide the means to plan,
prepare, execute and provide training feedback for Force on
Force (FOF) and Force on Target (FOT) training. Live
collective training exercises are characterized by the
following: g
•
Actual
soldier/vehicle
activity
on
terrain
under
simulated combat conditions. •
FOF weapon engagement with instrumented players via
Tactical Engagement Simulation and FOT with actual
targets and live fire. The live training domain includes all of the training
requirements and capabilities deployed by PEO STRI PM
Training Devices (TRADE) modernization strategy for the
Army’s Live Ground Maneuver Training Ranges. In order to
promote efficiencies for the Army live ranges across PM
TRADE
programs,
Goals
focus
on
capitalizing
standardization and interoperability efforts that address both
Government and Industry identified technology gaps, see
figure 2. The technology gaps require common live training
solutions to achieve the LT2 strategic mission to establish a
product line set of training systems that reduce total
ownership life cycle costs for the Army. The following
technology gap categories have been identified and have or
will be addressed by standardization efforts: y
•
Indoor Tracking requires high-fidelity, high-resolution
tracking data while indoors •
Indoor Tracking requires high-fidelity, high-resolution
tracking data while indoors BACKGROUND •
Scalability
of
communications
infrastructure
performance with increased range coverage and mobility INTRODUCTION product deployments: Combat Training Center Objective
Instrumentation System (CTC-OIS) to National Training
Center at Fort Irwin and Joint Range Training Center at Fort
Polk Fy08/Fy09; Homestation Instrumentation Training
System (HITS) Fort Bliss proof of concept Fy09; and
multiple Instrumentation Ranges sites to include Fort Carson
Fy09. Future product instantiation shall maximize existing
software core assets located in the LT2 Portal derived from
product instantiations and other common LT2 solutions
discussed within this paper. The U.S. Army Program Executive Office (PEO) Simulation
Training and Instrumentation (STRI) has established a Live
Training Transformation (LT2) product line approach to
developing a Family of Training Systems (FTS) that provide
the ground maneuver training range functions supporting
Army live and Joint training environments. The success of
the LT2 program strategy is dependent on defining standards
and initiatives derived to promote systematic reuse of
software and interoperability solutions for the products. Application of product line standards have matured reducing
life cycle development and sustainment costs through the
creation of common reusable architectures providing
integrated and interoperable training solutions for the
products deployable to Army Live Ranges. Standard
initiatives cover the following primary areas of technology
development: Tactical Engagement Simulation System
(TESS),
training
instrumentation
systems,
software
architecture, targetry, battlefield effects, and Joint Live-
Virtual-Constructive interoperability solutions. Approved
and
established
standards
promoting
reuse
and
interoperability include Common Training Instrumentation
Architecture (CTIA), OneTESS, Family of Army Systems
and Integrated Targetry (FASIT), Common Integrated Player
Unit Performance Specification Common Player Unit
Interface Control Document that includes MILES standard
compliance, and Consolidated Product Line Management
(CPM) strategy. The LT2 standards provide industry the
appropriate development guidelines and interface definitions
in order to maximize industry involvement in developing
product line solutions and providing advanced training
capabilities through technology insertion. This paper
provides an overview of how government and industry
worked together to establish the standard initiatives based on
common use cases and details the relevant solution sets
resulting in achievement of the PEO STRI mission and the
product line interoperability and reuse objectives. ABOUT THE AUTHORS Jorge Rivera is the LT2 Assistant Project Manager (APM) for LT2 at PEO STRI. His 21 years experience working in DoD
acquisition includes a strong software engineering background as the lead software engineer for several programs to include the
Joint Readiness Training Center – Instrumentation System and programmatic experience as the Project Director for Instrumented
Ranges and Urban Operations efforts. He earned his B.S. in Electrical Engineering from University of Puerto Rico in 1983 and
M.S.E.E. from Fairleigh Dickinson University, NJ in 1987. William Samper is currently assigned as the PEOSTRI Lead Engineer for the Common Training Instrumentation Architecture
(CTIA) and Live Training Transformation (LT2) Training Operations Center Command and Control (TOC2) program supporting
Homestation Instrumentation Training Systems (HITS) solutions. Experience includes 21 years of DOD acquisition supporting
project engineering and project management research, training systems, and product line solution efforts. He earned his B.S. in
Electrical Engineering from University of Florida in 1986. Barry Clinger is the Chief Technical Officer and VP of Engineering for Riptide Software, Inc. He is an experienced software
solutions expert who specializes in the design and architecture of large-scale distributed systems. Barry has over 18 years of
commercial, DoD, and NASA software development experience. He earned his B.S. in Computer Science from Eckerd College
in 1988 and his M.S. in Business from Kennedy Western University in 2003. He holds multiple software design and architecture
certifications from Sun and Microsoft. U.S. Government work not protected by U.S. copyright. 2008 Paper No.483Page 1 of 8 MILCOM 2008 Live Training Transformation (LT2) •
LVC Training, Test, and Joint Range end to end
application interoperability to include Common Data
Model Schema and fair fight correlation of models LT2 is an Army initiative to develop a live training range
product line that includes capabilities centered on a common
architecture, known as the Common Training Instrumentation
Architecture
(CTIA),
and
common
plug-and-train
components. The CTIA and LT2 components are used to
instantiate a specific modernized set of Live Ground
Maneuver Training Systems deployable by PEO STRI to the
Army live Combat Training Centers and Homestation
training environments, see figure 1. The product line strategy
is required to synergize training instrumentation, targets, and
tactical engagement simulation systems to ensure the
efficiency and effectiveness of training during peacetime,
mobilization, mission rehearsal, and in-theatre during
deployed military operations. Products are constructed using
a “family of components” approach, which maximizes
software reuse, provides common functionality, interfaces
and standards. The CTIA recently released its version 2.0
architecture on March 2008 and is supporting the following S •
Dismount and Tactical Engagement System Portable
Area Network Common Communications Solution that
provides seamless, wireless communications between
Player Units, communication infrastructure, and TES •
Standardized based Common Integrated Player Unit
(IPU) interchangeable between PUs and TES variants
that supports configurable multi-spectrum radios •
Battle Realism for Non-Line Of Sight engagements to
include munitions penetration effects •
Information Assurance support effort with Cross
Domain Solution to integrate Sensitive But Unclassified
with classified system secret high networks using lower
cost encryption solutions MILCOM 2008 What is LT2? What is LT2? The Live Training Transformation goal is to:
– Maximize Commonality
– Capitalize on Standardization
– Facilitate Interoperability
– Make the most of Component-based
Product Line Approach
• Maximizes Reusable Processes and
• Software Components
LT2 is a PM TRADE Modernization Strategy
The Live Training Transformation is a PM TRADE strategy which
– Promotes efficiencies across PM TRADE programs and Army Live Training
Ranges
– Aligns with and supports Army and DoD Training Transformation (T2)
– Provides dynamic, capabilities-based Live training solutions for the Army
and DoD in support of Warfighter needs
Figure 1. Live Training Transformation (LT2) Live Training Transformation •
Support common target integration solution to include
all existing deployed targets and modernized target
deployments for all FOT and FOF exercise types •
Support common target integration solution to include
all existing deployed targets and modernized target
deployments for all FOT and FOF exercise types Standardization evolution will be predicated on resolving the
common technology gaps identified in alignment with Army
Live Training Transformation modernization strategy in order
to maximize the total available set of Army training assets
used currently or planned for future deployment to the
Army’s Live Ground Maneuver Ranges. LT2 Standards Evolution One of the main cornerstones of the LT2 strategy is to
capitalize on standardization to maximize commonality and
facilitate interoperability, and subsequently promote cost
savings across PM TRADE acquisitions. Figure 1. Live Training Transformation Technology Gaps
Technology Gaps
AAR
X
O/C
x
Live OPFOR
Targets
OPFOR
OC/TAF
Virtual/Constructive Wrap Around
TE ~ COE (JIIM)
Role
Players
Battlefield Effects
Live/Virtual/Constructive
Virtual/Constructive Wrap Around
TE ~ COE (JIIM)
BLUE C4I
x
Urban Ops Area
Indoor
Tracking
Information
Assurance
DMT / TES
PAN
Non-Contact
Hit Sensors
Live Players
Live DMT
TES
ROC / TOC2
Interoperability
Scalability / Mobility
Battlefield Realism
Common IPU
Adaptive
Behavior Tgts
Figure 2. LT2 Domain Problem Space Information
Assurance Indoor
Tracking Adaptive
Behavior Tgts Non-Contact
Hit Sensors Scalability / Mobility Common IPU Figure 2. LT2 Domain Problem Space Live Training Transformation (LT2) Technology Gaps
Addressed standards, interfaces, processes, and requirements that directly
support the LT2-FTS programs. Figure 3 shows CPM
strategy to consolidate all LT2-FTS Post Product Software
Support and Post Deployment Software Support within a
single management structure and organization to maintain the
product line integrity of the LT2-FTS programs. A definition of “standardization,” in the context related to
technologies and industries, is “the process of establishing a
technical standard among competing entities in a market,
where this will bring benefits without hurting competition.”
PM TRADE’s goal of standardization centers around a
universally agreed upon set of guidelines that support
interoperability efforts within PM TRADE, across PEO STRI
and the Army. Within this context, PM TRADE will follow a
systematic Consolidated Product Line (CPM) management
approach which uses a centralized repository, known as the
LT2 Portal, to manage, develop, and sustain its architectures, LT2 Standardization Process LT2 uses multiple approaches in working with industry and
stakeholders
to
establish
standardized
processes
for
development and interoperability. One of the methods
successfully employed is, Industry Days, where Government
comes together with industry to describe the current live MILCOM 2008 processes. In addition, approved a concept of operations that
establishes the PM TRADE management structure and
processes necessary to execute the LT2 strategy and
standardization efforts across all PM TRADE programs, and
the new live training capabilities defined in the approved Live
Training Transformation Family of Training Systems (LT2-
FTS) Initial Capabilities Document (ICD). The LT2 ConOps
provides for a Technical Advisory Group (TAG) structure to
bring stakeholders together in evolving new standards and
providing the necessary recommendations to lower and higher
level management via either the Product Advisory Group
(PAG) or Board of Directors (BOD) 2006). Documented
standards and processes are referenced within the PM
TRADE acquisition process to foster reuse and reduce total
life cycle costs for new products. Examples of standards
referenced in the acquisition process include the following: processes. In addition, approved a concept of operations that
establishes the PM TRADE management structure and
processes necessary to execute the LT2 strategy and
standardization efforts across all PM TRADE programs, and
the new live training capabilities defined in the approved Live
Training Transformation Family of Training Systems (LT2-
FTS) Initial Capabilities Document (ICD). The LT2 ConOps
provides for a Technical Advisory Group (TAG) structure to
bring stakeholders together in evolving new standards and
providing the necessary recommendations to lower and higher
level management via either the Product Advisory Group
(PAG) or Board of Directors (BOD) 2006). Documented
standards and processes are referenced within the PM
TRADE acquisition process to foster reuse and reduce total
life cycle costs for new products. Examples of standards
referenced in the acquisition process include the following: training problem domain space and state the prioritized non-
interoperability or stovepipe issues that exist at the Army live
ranges between deployed simulation systems. In addition
LT2 vision and mission strategies are briefed to industry
along with current plans to deliver future modernized training
devices in order to educate training needs to industry and
encourage adoption of LT2 and CTIA capabilities and
strategies (i.e. LT2 CPM concept). LT2 Standardization Process Fu nctional Cap ab ility Groups
Planning
SYSCON
EXCON
Data
Collection
BattleField
Realism
T AF
As of 11 Jun 0 7 (V1.4)
After Action
Review
Field Camera Ctrl
Vehicle Video Ctrl
Video System
Suite
Air Warrior
Digital Tactical
Monitoring
Air Defense
Artillery Fire
AWES
DIS/HLA Gateway
Fire finder Radar
IEWTPT Gateway
Ranges Pairing
Processor
Wrap-around
Sim Gateway
Battle Roster
DCP Editor
Range Data
Editor
Rolling
Combat Power
Alarm & Alerts
Event Generator
Pairing Processor
ADRM
STM
CTIA Explorer
Roles &
Perm issions
Obstacle Report
Ground Player
Instr. Controller
Instr. Issue &
Recovery (IIR)
Contact Report
Weather Station
Processor
Observation
Recording
EXCON
CAS Mission
Editor
2D Map
E xercise Manager
2D Map Lite
Ad Hoc
Query Tool
InBox
Handheld Target
Control
Jamm ing
Config. View
Participant
Definition
LT2 Compon en ts
System Compositions
Combat Training Centers
(CTCs)
Home Station
Deployed
Architecture
Infrastru cture Layer
C++, C# & Java
Object Model
CTIA
Services
Data Distribution
Manager
Operating
System
Wired/Wireless
Co mmunicatio ns
LT2 Datab ase
Preferences Editor
Tactical Net
Selector
Data Access
Layer
Exercise Tree
FBCB2 DB
GPS Processor
Playback
Scenario
Controller
Scoring Processor
SYSCON
Voice Tactical
Monitoring
LSF GUI
Fram ework
Infrastructure
Scenario
Development Tool
4/5
5/7
13/18
8/13
8/10
4/7
6/6
- Future Component
Aviation P layer
Instr. Controller
UO Device
Controller
Ground Player
Inst. Server
Non- Instr. Range Services
Force
Structure
Bookmark Tool
Event Log
Universal
Target Controller
Exercise Builder
LT2 GUI
Fram ework
HITS ESC
3D Map
Player Unit
- CTIA Gateway
HITS ESC
- Not Used by Objective HITS
- Future HITS Component
- Researching
Universal
Target Controller
Target Event
Processor
Player Chiclets
Embedded
Battle Roster
System Wizard
P layer User List
Functional Capability Groups
P lanning
SYSCON
EXCON
Data
Collection
BattleField
Realism
TAF
As of 06 Sep 07 (V1. 5.1)
After Action
Revi ew
Fie ld Camera C trl
Vehic le Video
Control
Video System
Suite
Ai r Wa rrior
Digital Tac tic al
Monitoring
Air Defense
Artille ry Fi re
AWES
DIS/HLA Ga tew ay
Fire finder Ra da r
IEWTPT Gatewa y
Ra nges Pairing
Proce ssor
Wrap-a round
Sim Ga tew ay
Ba ttle Roster
DCP Editor
Range Data
Editor
Rol ling
Combat Powe r
Ala rm & Alerts
Eve nt Ge ne ra tor
Pa iri ng Proce ss or
ADRM
STM
CTIA Ex plorer
Roles &
Permissi ons
O bs tac le Re port
Ground Pl aye r
Instr. LT2 Standardization Process Control ler
Instr. I ssue &
Re covery (IIR)
Contac t Re port
Wea ther Sta tion
Li te
O bs ervation
Rec ordi ng
EXCON
CAS Mis sion
Editor
2D Ma p
Exe rc ise Ma na ge r
2D Ma p Lite
Ad Hoc
Query Tool
InBox
Handheld Targe t
Control
J ammi ng
Config. Vie w
Partic ipa nt
Defi nition
LT2 C om pone nts
S ys te m C omposi ti ons
Combat Training Centers
(CTCs)
Home Station
Deployed
A rc hitec ture
I nfras truc ture La ye r
C ++, C# & J av a
Obje c t M ode l
CTIA
S erv ic es
Da ta D is tribution
Mana ger
O pera ti ng
Sys te m
W ire d/Wi re le s s
Communi ca ti ons
LT2 D ata bas e
Pre ference s Editor
Tactica l Ne t
Se lec tor
Da ta Acce ss
La yer
Exe rc ise Tree
FBCB2 D B
G PS Proc es sor
Pla yba ck
Sc enario
Controll er
Scoring Proce ssor
SYSCON
Voic e Tac tic al
Monitoring
LSF GUI
Framew ork
I nfras truc ture
Sce na ri o
De ve lopment Tool
3/5
1/ 7
8/18
3/15
0/ 10
1/7
1/ 6
- Future C omponent
Ground Pl aye r
Inst. Serve r
Non- Instr. Se rv ice s
Force
Structure
Bookmark Tool
Ev ent Log
Unive rs al
Targe t Controll er
Exe rc ise Builder
LT2 GUI
Fra me work
17/ 68
3D Ma p
IR
IR
IR
IR
IR
IR
FAS IT
FASIT
IR
IR
IR
IR
IR
IR
IR
IR
IR
IR
IR
IR
IR
FAS IT
FAS IT
IR
Ta rget Event
Proce ssor
CTC-O IS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
IR
CTC
-OI S
CTC-OIS
C TC-O IS
CTC-OIS
C TC-O IS
CTC-OIS
CTC-OI S
IR
CTC-OIS
C TC-O IS
Cause & Effe ct
Tool
Orders Ca pture
H ITS Pla yer Unit
- CTIA Gatewa y
Av Pl ayer Instr. LT2 Standardization Process Ctrl
UO Device Ctrl
CTC-OIS
CTC
-OIS
H ITS
LT2-FTS CPM Strategy
LT2-FTS CPM Strategy
ETC
CPM
Repository
Fu nctional Cap ab ility Groups
Planning
SYSCON
EXCON
Data
Collection
BattleField
Realism
T AF
As of 11 Jun 0 7 (V1.4)
After Action
Revi ew
Fi el d Camera Ctrl
Vehicle Video Ctrl
Video System
Suite
Air Warrior
Digital Tactical
Moni toring
Air Defense
Artill ery Fire
AWES
DIS/HLA Gateway
Fi re finder Radar
IEWTPT Gateway
Ranges Pairing
Processor
Wrap-around
Si m Gateway
Battl e Roster
DCP Editor
Range Data
Editor
Rol ling
Combat Power
Al arm & Alerts
Event Generator
Pairing Processor
ADRM
STM
CTI A Explorer
Roles &
Perm issions
Obstacl e Report
Ground Player
Instr. Controll er
I nstr. Issue &
Recovery (IIR)
Contact Report
Weather Station
Processor
Observation
Recording
EXCON
CAS Mi ssion
Edi tor
2D Map
E xercise Manager
2D Map Lite
Ad Hoc
Query Tool
InBox
Handhel d Target
Control
Jamm ing
Config. Vi ew
Participant
Defini tion
LT2 Compon en ts
System Compositions
Combat Training Centers
(CTCs)
Home Station
Deployed
Architecture
Infrastru cture Layer
C++, C# & Java
Object Model
CTIA
Services
Data Distribution
Manager
Operating
System
Wired/Wireless
Co mmunicatio ns
LT2 Datab ase
Preferences Editor
Tactical Net
Selector
Data Access
Layer
Exercise Tree
FBCB2 DB
GPS Processor
Pl ayback
Scenario
Control ler
Scori ng Processor
SYSCON
Voi ce Tactical
Moni toring
LSF GUI
Fram ework
Infrastructure
Scenario
Development Tool
4/5
5/7
13/ 18
8/13
8/10
4/7
6/6
- Future Component
Aviation P layer
Instr. Controll er
UO Device
Controller
Ground Player
Inst. Server
Non- Instr. LT2 Standardization Process Range Services
Force
Structure
Bookmark Tool
Event Log
Uni versal
Target Control ler
Exercise Buil der
LT2 GUI
Fram ework
HITS ESC
3D Map
Player Uni t
- CTIA Gateway
HITS ESC
- Not Used by Objective HITS
- Future HITS Component
- Researching
Uni versal
Target Control ler
Target Event
Processor
Player Chiclets
Embedded
Battl e Roster
System Wi zard
P layer User Li st
LVC
Joint Ranges
Functional Capability Groups
Planning
SYSCON
EXCON
Data
Collection
BattleField
Realism
TAF
As of 20 April 07 (V1.3 )
A fter Action
Review
Fiel d Camera Ctrl
Vehicl e Video Ctrl
Video System
Suite
Air Warri or
Digital Tactical
Monitoring
A ir Defense
Artillery Fire
AWES
DIS/H LA Gateway
Fi re finder Radar
IEW TPT Gateway
Ranges Pairing
Processor
Wrap-around
Sim Gateway
DCP Editor
R ange Data
Editor
Rolli ng
Combat Power
Alarm & Alerts
Event Generator
Pairing Processor
ADRM
STM
CTIA Expl orer
Roles &
Permi ssions
Obstacle Report
Orders Capture
B ookmark Tool
Instr. Issue &
Recovery (I IR)
Weather Stati on
Processor
Observation
Recording
EXCON
CAS Mission
Editor
U ni versal
Target Control ler
2D Map
Exercise Manager
2D Map Li te
Ad Hoc
Query Tool
InBox
Cause & Effect
Tool
Handheld Target
Control
Jamming
Config. View
Partici pant
Definition
System C ompositi ons
Combat Training Centers
(CTCs)
Home Station
Deployed
A rchitecture
I nfrastructure Layer
C ++, C# & Java
Object M odel
CTI A
Services
Data Di stribution
Manager
Operati ng
System
W ired/W ireless
C omm unicati ons
GU I
Fram ework
LT2 Database
Preferences Editor
Tactical N et
Selector
Data Access
Layer
Exercise Tree
FB CB2 DB
GPS Processor
Pl ayback
Scenari o
C ontroller
Scoring Processor
SYSCON
Voice Tactical
Monitoring
Exercise B ui der
Infrastructure
Scenario
Development Tool
3/5
6/7
13/1 8
6/1 5
8 /10
4/ 7
0/6
- Future Component
Contact R eport
LSF GUI
Framework
Non-Instrumented
Range Services
Force
Structure
Battl e R oster
3D Map
HITS Player Uni t
- CTIA Gateway
- Component Currently Available
40 /68 [59%]
LT2 C omponents
CTC-OIS : Denotes CTC-OIS is Lead Develope r [21/40]
CTC- OIS
CTC- OIS
IR
CTC- OIS
CTC- OIS
CTC- OIS
CTC- OIS
IR
IR
CTC- OIS
CTC- OIS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-O IS
CTC-OIS
FASIT
CTC-OIS
CTC-O IS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
Target Event
Processor
CTC-OIS
LT2 GUI
Framework
Ground Player
I nstr. LT2 Standardization Process C ontroller
Av Play er I nst r. Ctrl
UO Device Ctrl
Ground Player
Inst. Server
Event Log
Functional Capability Groups
Planning
SYSCON
EXCON
Data
Collection
BattleField
Realism
TAF
As of 20 April 07 (V1.3 )
After Action
R evi ew
Fi eld C amera Ctrl
Vehi cle Video C trl
Vi deo System
Suite
A ir Warrior
Di gi tal Tacti cal
Monitoring
Air D efense
Arti llery Fire
AW ES
D IS/HLA Gateway
Fire finder R adar
IEWTPT Gateway
Ranges Pairing
Processor
Wrap-around
Sim Gateway
Battl e Roster
DCP Editor
R ange D ata
Editor
Rolli ng
C ombat Power
Alarm & Al erts
Event Generator
Pairing
Processor
ADR M
STM
CTIA Explorer
Roles &
Permissions
Obstacle Report
Orders Capture
Bookmark Tool
Instr. Issue &
Recovery (I IR)
Weather Stati on
Li te
Observation
Recording
EXCON
CA S Mission
Editor
2D Map
Exercise Manager
2D Map Lite
Ad Hoc
Query Tool
I nB ox
Cause & Effect
Tool
H andheld Target
Control
Jamming
Config. Vi ew
Participant
D efiniti on
LT2 Components
System Compo sitions
Combat Training Centers (CTCs)
Home Station
Deployed
Architecture
Infrastru cture Layer
C++, C# & Java
Object Model
CTIA
Services
Data Distribution
Manager
Operating
System
Wired/Wireless
Communications
GUI
Framework
LT2 Database
Preferences
Edi tor
Tactical Net
Selector
Data A ccess
Layer
Exercise Tree
FBC B2 DB
GPS Processor
Playback
Scenario
Control ler
Scoring Processor
SYSCON
Voice Tactical
Monitoring
Infrastru cture
Scenario
Development Tool
3 /5
3/7
8/ 18
7/1 5
2/1 0
4/7
3/6
- Future Component
Contact R eport
LSF GUI
Framework
N on-Instrumented
Services
Force
Structure
Universal
Target C ontroller
3D Map
HITS Player Unit
- CTIA Gateway
- Component Cu rrently Available
30 /68 [ 44 %]
IR
IR
IR
IR
IR
IR
IR
IR
IR
IR
IR
IR
IR
Exercise Buil der
IR
IR
IR
IR
Event Log
IR
IR
Target Event
Processor
IR
IR : Denotes IR is Lead Developer [21/30]
CTC -OIS
CTC-OIS
CTC -OIS
CTC -OIS
C TC-OIS
FASIT
CTC-OIS
CTC-O IS
Ground Player
Instr. Control lerIR
Ground Pl ayer
I nst. Server IR
LT2 GU I
Framework
Av iation Play er
Instr. Controller
UO Devic e
Controll er
CPM focus is PPSS/PDSS Configuration Management and
product line integrity of LT2-FTS
Functional Capability Groups
P lanning
SYSCON
EXCON
Data
Collection
BattleField
Realism
TAF
As of 06 Sep 07 (V1. LT2 Standardization Process 5.1)
After Action
Revi ew
Field Camera C trl
Vehicle Video
Control
Video System
Suite
Ai r Warrior
Digital Tactical
Monitoring
Air Defense
Artillery Fi re
AWES
DIS/HLA Gatew ay
Fire finder Radar
IEWTPT Gateway
Ranges Pairing
Processor
Wrap-around
Sim Gatew ay
Battle Roster
DCP Editor
Range Data
Editor
Rol ling
Combat Power
Alarm & Alerts
Event Generator
Pairi ng Processor
ADRM
STM
CTIA Explorer
Roles &
Permissi ons
O bstacle Report
Ground Pl ayer
Instr. Control ler
Instr. I ssue &
Recovery (IIR)
Contact Report
Weather Station
Li te
O bservation
Recordi ng
EXCON
CAS Mission
Editor
2D Map
Exercise Manager
2D Map Lite
Ad Hoc
Query Tool
InBox
Handheld Target
Control
Jammi ng
Config. View
Participant
Defi nition
LT2 C om ponents
S ystem C omposi ti ons
Combat Training Centers
(CTCs)
Home Station
Deployed
A rchitecture
I nfrastructure Layer
C ++, C# & Java
Object M odel
CTIA
S ervices
Data D istribution
Manager
O perati ng
System
W ired/Wi reless
Communi cati ons
LT2 D atabase
Preferences Editor
Tactical Net
Selector
Data Access
Layer
Exercise Tree
FBCB2 D B
G PS Processor
Playback
Scenario
Controll er
Scoring Processor
SYSCON
Voice Tactical
Monitoring
LSF GUI
Framew ork
I nfrastructure
Scenari o
Development Tool
2/5
7
4/18
15
10
7
4/6
- Future C omponent
Ground Pl ayer
Inst. Server
Non- Instrumented
Servi ces
Force
Structure
Bookmark Tool
Event Log
Universal
Target Controll er
Exercise Builder
LT2 GUI
Framew ork
68
3D Map
IR
IR
IR
IR
IR
IR
FASIT
FASIT
IR
IR
IR
IR
IR
IR
IR
IR
IR
IR
IR
IR
IR
FASIT
FASIT
IR
Target Event
Processor
CTC-O IS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
CTC-OIS
IR
CTC
-OI S
CTC-OIS
C TC-O IS
CTC-OIS
C TC-O IS
CTC-OIS
CTC-OI S
IR
CTC-OIS
C TC-O IS
Cause & Effect
Tool
Orders Capture
H ITS Player Unit
- CTIA Gateway
Av Pl ayer Instr. Ctrl
UO Device Ctrl
CTC-OIS
CTC
-OIS
H ITS
Warfighter Focus
Field Support
LT2 Portal
& Components
Figure 3. LT2-FTS CPM Strategy renced in the acquisition process include the following: •
Component Agreement Template contains an overall
description of the component, interface definition, and
its purpose and provides context for the Industry vendor. •
Human Computer Interface Style Guide leads application
developers in the design and implementation of
compliant applications, displays, controls and visual
components. Figure 3. LT2-FTS CPM Strategy With the establishment of the web-based LT2 portal as a
CPM repository, LT2 has created collaboration working
groups with participation from Government and Industry that
support standards evolution and common solutions within the
following areas: LT2 Developer’s Guide provides the background,
procedures, and required reference material for a
software developer or organization to successfully craft
software using the existing assets of the LT2 Product
Line and to submit potential assets for future LT2
Product Line inclusion. LT2 Developer’s Guide provides the background,
procedures, and required reference material for a
software developer or organization to successfully craft
software using the existing assets of the LT2 Product
Line and to submit potential assets for future LT2
Product Line inclusion. •
Common Component Working Group. The goal of this
group is to discover and foster the evolution of
components that can be reused across the Product Line. •
Architecture Working Group facilitates the coordination
of the CTIA Architecture Change Process LT2 INTEGRATED AND INTEROPERABLE
SOLUTIONS •
Test Tools group provides available tools used to test the
CTIA architecture and LT2 components. •
Development and review of the Instrumented Player Unit
Performance Specification (IPUPS). LT2 Standardization Process A common framework for Human Control
Interface (HCI) design is provided, enabling all
applications to appear and operate in a reasonably
consistent manner. Design guidelines in the form of a
style guide provide three major benefits: higher
productivity, reduced development time, and reduced
training time Figure 3. LT2-FTS CPM Strategy Common Training Instrumentation Architecture (CTIA) The flexibility in scaling of the system is
achieved by the “plug and play” design of the architecture. Figure 4 shows the data distribution, including plug and play
design. Common Training Instrumentation Architecture (CTIA) •
LT2-FTS product working groups support analysis of
software requirements and development of software
components developed new, modified, or reused for
future reuse by specific programs (i.e. HITS). CTIA is the product line architecture that provides
commonality across training instrumentation systems and
interoperability across Live, Virtual, Constructive (LVC), and
Joint training systems. It consists of standards and protocols
to be used by systems developers and is the core software
component of the LT2-FTS. CTIA is a component-based,
domain specific, product line architecture that enables the
LT2 strategy and leverages the high degree of commonality of
requirements among the products deployed to CTCs and
home stations. This emphasis on commonality by the CTIA
will improve the quality of training while significantly
reducing development, logistics, training, and maintenance
costs. CTIA and associated processes will be the basis for
subsequent product line members to leverage the product line
architecture and common software components. •
LVC Integration and Interoperability assesses the
existing Service Oriented Architecture type architectures
for support of developing a prototype LVC strategy
Development, documentation, and maintenance of ICDs
and the architecture for a common messaging format and
protocol between PUs and CTIA services. •
LT2 Portal Content and Common Portal Engine
standards •
Future Army System of Integrated Targets (FASIT)
standards between PEO STRI, Industry, and other
interested parties. CTIA implements a set of domain-specific services to support
Army Live Training. These can be thought of as logically
centralized, but potentially, physically distributed services. The CTIA Services component provides a set of APIs LT2 has created and documented standard processes for
approving architecture changes, common component changes
and development standards using the Portal as a formal
review and sign-off mechanism for evolving standards and 2007 Paper No. 7200 Page 4 of 8 MILCOM 2008 (defined using CORBA IDL and XML) that interact with LT2
components, allow for the interaction between other
components, and provide common LT2 database schema
implemented in Structured Query Language (SQL). For ease
of use object model frameworks are provided for the primary
languages (C++, Java, C# and .NET) that wrap services and
provide easier programmatic interface. The LT2 Product Line uses communication standards to
integrate component based elements that can run distributed
across many hardware platforms for large scale exercises or
logically combined across a few hardware platforms for small
scale exercises. CTIA Standard Solutions Range File Server
Data Distribution within CTIA
Data Distribution within CTIA
JDBC/ADO.Net
over TCP/IP
CTIA Services
Exercise Independent/
Exercise Specific
CORBA RMI
over TCP/IP (IIOP)
LT2 Databases
Exercise
DB
DCP
DB
Asset
DB
DBMS (Oracle)
SQL
CORBA
Unicast
CORBA IDL defined Event Data
over UDP Unicast
CORBA
LT2 Plug And Play
Components
(Processors, Instrumentation)
CORBA
Multicast
Multicast
Application
Server
(JBOSS)
CORBA IDL defined Stream Data
over UDP Multicast
File
System
WEBDAV
WEBDAV/HTTP
over TCP/IP
XML
Data
Model
Other Ranges/
Other Services
TENA LROM
TENA TDL
(CORBA RMI + UDP Multicast)
over TCP/IP
TENA LROM
CTIA
Database
Services (EJB)
HLA/
DIS
Simulations
Tactical/C4I
Systems
TESS
Devices
Targetry
Legend
CTIA Services Components
COTS Products
LT2 Components
CTIA Architecture-Defined Interfaces
Other Applications/Systems
Data Exchange
BES
Devices
Position
Reporting
Devices
Document Server
Audio/
Video
CTIA Flexible Features
defined in XML
SLP Agent
(Open SLP)
SLP
SLP (Service Location) data
over IP/UDP Unicast and Mutlicast
Figure 4. CTIA Data Distribution Standards CTIA provides a compliance definition that specifies the
requirements for LT2 components to be assessed as CTIA-
compliant for systemic or opportunistic reuse by the Family of
Training Systems. Additionally, CTIA specifies the
requirements for a Product to be CTIA Product compliant. A
training range consists of many systems and components. There will be systems that will not be CTIA Product
compliant; however they can interoperate with LT2 FTS
through CTIA Component compliance. CTIA components
which need to operate within LT2-FTS will have differing
integration needs and different levels of compliance. A CTIA compliant component is developed to conform to
CTIA architecture object model frameworks and interface
specification requirements to support reuse and instantiation
within LT2 products using CTIA as its core architecture. The
CTIA Product Line Architecture Specification (PLAS),
Product Line Architecture Framework (PLAF), and associated
processes will be used to realize CTIA-based LT2 products
composed of plug and train common and unique LT2
components. . HLA/
DIS
i
l ti Legend The component agreement serves as an agreement between
the component developer, the users, and PEO STRI LT2
management organization. Higher levels of CTIA-compliance Figure 4. CTIA Data Distribution Standards Figure 4. CTIA Data Distribution Standards g
g
g
p
realize greater reusability across the product line because: •
Providing Component Agreements specifies the
requirements, interfaces, behaviors and dependencies of
components, which allows lead system integrators the
selection of appropriate components for reuse within
their product. Future Army Systems of Integrated Targets (FASIT) LT2 has created FASIT common standards to support
component plug and play technology insertion relative to
integration of targetry and range devices used to support live
Force on Target training objectives as follows: •
Providing
Component
Agreements
also
fosters
systematic reuse by allowing the product line to
collaborate on the development of requirements for
components during design and development phases, so
that components can be designed to be reused across
products and ranges. •
Provides Army Common Standards for Live Fire
Training Devices •
Includes Protocols for Target/Device Control in Training
Circular 25-8 that includes Urban Operations,
Instrumented Ranges, CTCs and home stations •
Adherence to the interfaces defined in the CTIA PLAS
supports reusability by ensuring that component
interfaces are defined and managed by the product line. •
Allows mixing of different vendor’s products (also
supports range non-FASIT RETS targetry that are
obsolete) on a single range under a single control system
(TRACR) •
Adherence to the LT2 GUI Style Guide facilitates
reusability by ensuring common approaches to user
interface
design
across
independently
developed
applications. FASIT has requirements to provide the common standards,
interfaces, and protocols to develop a CTIA compliant
Common Target Controller comprised of a Universal Target
Controller (UTC) and the Targetry Range Automated Control •
Use of CTIA Frameworks reduces development and
maintenance costs by encapsulating architecture changes
and providing a reusable library of common software. 2007 Paper No. 7200 Page 5 of 8 MILCOM 2008 exercise to crew/squad drills to prepare the Force to
execute its wartime mission. The OneTESS will
support the execution of proper engagement procedures;
provide simulated weapon system's accuracy and
effects; and adjudicate results of engagements. Creation
of an OneTESS Standard is necessary to establish and
maintain configuration control over functionality and
ensure hardware and software interoperability with
MILES variants and fielded equipment. Standard will
eliminate variation in results that discredit the validity
of a training exercise or test event. OneTESS will also
provide modeling, troubleshooting, and conformance
tools to assist in implementation of the standard. and Recording System (TRACR) for reuse by LT2 products. FASIT allows LT2 products to support live fire exercises
(LFXs) for individual and crew served weapon skill
qualification and sustainment, and, collective training events
at local training areas, combat training centers, and in tactical
force projection environments. One Tactical Engagement Simulation Systems (OneTESS) The OneTESS is a product that provides a common TES
architecture for integrating current and future vehicles, and
weaponry found in the heavy, infantry and Stryker Brigade
Combat Teams and OPFOR. The components of OneTESS
will be compliant with the Common Training Instrumentation
Architecture (CTIA) that is used throughout the LT2-FTS. OneTESS will use actual or simulated weapons systems
equipped with tactical engagement simulators to replicate
weapons’ effects and battle damage. OneTESS will operate
predominantly in conjunction with a Combat Training Center
(CTC) or Homestation Instrumentation Training System
(HITS) to provide the common TES capability in an exercise. OneTESS also will have a standalone mode that provides the
exercise management function as well as the TES capability
to support company-size homestation training exercises. The IPU provides all live entities networked in the training
FOF or FOT exercise the capability to process the necessary
casualty assessment information, and communicates this
information back to the Range Operating Complex. The four
main types of players in a live training exercise that require
the functions of an IPU are: (1) the Dismount, or individual
Soldier, (2) the Vehicle, to include all ground and air
vehicles, (3) Targets, and (4) Observer Controller. The IPU
consists of three main modules: (1) the Instrumentation
Processor (IP), (2) the Platform Interface Processor (PIP), and
(3) the TESS Processing Unit (TPU). OneTESS Increment 1 will enable the Real Time Casualty
Assessment of live FOF and FOT training with vehicles and
weapons found in the Brigade Combat Teams and in the
OPFOR at the Combat Training Centers. It will provide non-
line-of sight (NLOS) capability to adjudicate employment of
indirect fire/area effects weapons as well as line of sight
(LOS) capability for direct fire weapons. The focus of training
is at the Brigade Combat Team (BCT)-level and below. Increment 1 of OneTESS uses the geographic position,
weapon orientation vector and trajectory computation of
geometric-pairing for simulating indirect fire
engagements and laser generators/detectors to simulate
direct fire engagements thus providing a comprehensive
system to fairly adjudicate live training and testing
exercises. OneTESS components will be integrated
across the live training domain and support all phases in
the training cycle at all echelons from a joint level OneTESS Increment 1 will enable the Real Time Casualty
Assessment of live FOF and FOT training with vehicles and
weapons found in the Brigade Combat Teams and in the
OPFOR at the Combat Training Centers. Future Army Systems of Integrated Targets (FASIT) FASIT has developed LT2
common components to provide realistic friendly, neutral, and
threat targets and weapon effect simulation; define targetry
range (Range Data Editor); target scenario preparation and
execution (Scenario Development Tool); target status; data
collection; automated maintenance support; test suite; and
after exercise execution, FASIT supports training preparation,
presentation, and feedback. FASIT supports Interface
Control Documents for following: battle effects and targets;
battle position sensors; small range ERETS protocol adaptor
software; and LT2 Instrumented PU systems. Common Integrated Player Instrumentation (IPU) FASIT has allowed integration of multiple vendor hardware
to work seamlessly with LT2 products. In addition FASIT has
provided a solution to replacing obsolete RETS range targetry
with FASIT compliant targets at a significant cost savings to
the Army and has also supported a retrofit of small non-
instrumented ranges obsolete targets with FASIT compliant
interoperability to LT2 products. This Integrated Player Unit Performance Specification
(IPUPS) establishes the performance and validation
requirements for the Instrumented Player Unit (IPU) for
various programs. The IPUPS standard will foster future
design efforts to replace the current set of legacy IPU or Data
Communications Interface (DCI) designs. This new family of
live training IPU design will provide the same functions as
before, and will also be based on a modular, open architecture
approach that accommodates simulating new weapon systems,
stresses “train as you fight” concepts, incorporates the Future
Force requirements, provides enhanced training data
collection and After Action Review (AAR) capabilities,
power requirements, Personal Area Network (PAN)
requirements, verification methods, independence of radio
communications protocols, performance measures, common
message formats, common connector standards, improved
maintenance, and facilitates interoperability and technology
insertion. The goal is for this IPU design to incorporate
current Tactical Engagement Simulation Systems (TESS)
interoperability as well as provide a base for the development
of future TESS and IPU interoperability evolution as LT2
migrates to future OneTESS Standard. 2007 Paper No. 7200 Page 6 of 8 One Tactical Engagement Simulation Systems (OneTESS) It will provide non-
line-of sight (NLOS) capability to adjudicate employment of
indirect fire/area effects weapons as well as line of sight
(LOS) capability for direct fire weapons. The focus of training
is at the Brigade Combat Team (BCT)-level and below. Increment 1 of OneTESS uses the geographic position,
weapon orientation vector and trajectory computation of
geometric-pairing for simulating indirect fire
engagements and laser generators/detectors to simulate
direct fire engagements thus providing a comprehensive
system to fairly adjudicate live training and testing
exercises. OneTESS components will be integrated
across the live training domain and support all phases in
the training cycle at all echelons from a joint level The PU Interface Control Document (ICD) mitigates the
challenge of having stovepipe communications systems and
PU networks by defining a single interface method with
common XML protocols for passing messages to all types of
Player Units (legacy and common IPU), and for facilitating
interoperability between all types of Player Units and the
Common Training Instrumentation Architecture (CTIA)
services. Live Training products that are part of the LT2 2007 Paper No. 7200 Page 6 of 8 MILCOM 2008 today within industry, however, how to consistently utilize
these capabilities in LT2 applications was the challenge. today within industry, however, how to consistently utilize
these capabilities in LT2 applications was the challenge. Family of Training Systems (LT2-FTS) may develop or
extend software modules (PU gateways and common PU
controller) used to integrate PUs previously developed by
utilizing a “plug-and-play” approach via CTIA Services,
CTIA compliance definitions, and PU ICD as design
guidance. The PU ICD standard provided the following
benefits: A video SOA was established to allow a consistent
mechanism for accessing and manipulating video within LT2,
while
allowing
flexibility
in
the
underlying
video
technologies actually being used. By specifying how to
describe video sources and how to retrieve them without
mandating specific technology choices provides LT2
applications the flexibility to select the type of video
hardware and software that makes sense for that application. The video SOA also provides the means to exchange video
services between applications, and provide additional
incremental functionality to be added as necessary. For
instance, one defined video service may simply provide video
feeds from known geographic locations within a building. One Tactical Engagement Simulation Systems (OneTESS) • Camera Position
• Source Metadata
• Play List
• Stream Collection
• Camera Control
• Live Streams
• Recording
• Video Query
• Video Work Station • Camera Position
• Source Metadata
• Play List
• Stream Collection
• Camera Control
• Live Streams
• Recording
• Video Query
• Video Work Station • Video Work Station Player Unit Interface with CDS View
Player Unit Interface with CDS View
• A common PU standard, based on existing capabilities, provides a baseline to be used as a
starting point for transition/migration to “OneTESS” common PU solution
• Allows single CDS guard certification when introducing different PU networks
• Provides automatic compliance with use of LT2 PU ICD standard Player Unit Interface with CDS View
Player Unit Interface with CDS View Conclusions and Summary The LT2 Standardization efforts and Consolidated Product
Line Management strategy provides the following benefits
and lessons learned: •
CPM provides a single management mechanism to
consolidate all LT2-FTS sustainment and enhancement
development efforts fostering common live training
solutions while lowering total ownership life cycle costs
across programs and mitigating potential cost, schedule
and performance risks •
CPM allows for consolidated lab and test resources at a
single facility site reduces overhead and Integration and
Test costs, reduces IAVA management and IA
certification costs across LT2-FTS, reduces
configuration and inventory management costs, and
provides for help desk support • A common PU standard, based on existing capabilities, provides a baseline to be used as a
starting point for transition/migration to “OneTESS” common PU solution
• Allows single CDS guard certification when introducing different PU networks
• Provides automatic compliance with use of LT2 PU ICD standard Figure 5. Common IPU Interface Standard •
Quality of service and products are increased through
testing and resolving issues once and deployment of
common solutions to all programs and training sites Video Service-Oriented Architecture (SOA) One Tactical Engagement Simulation Systems (OneTESS) A
second video service may combine known locations from live
players as they traverse that building, with the known
locations of the video sources to provide automatic video
tracking of live players as they move throughout a building. This is one example how SOA-based technology can grow to
flexibly accommodate new requirements without permutation
to existing services. The current existing video SOA services
include: •
Allowed reuse for LT2 Player Unit Common Message
Set. Instead of creating multiple stovepipe custom
communications mechanisms for different player types,
this ICD establishes a standard for the LT2 domain. A
single LT2 component can be developed to work with all
PU types. Helper software modules can be created and
disseminated to assist with the development of new PU
gateways. •
Provided automatic compliance when using the LT2
Component. When developers utilize this ICD and the
associated components and helper software modules, the
developers can be assured of LT2 compliance. •
Provided a simple path for trusted guard certification. XML is a mature and well-known software
communications mechanism and allows for relatively
easy and fast Information Assurance (IA) accreditation
for Cross-Domain Solutions. Furthermore, even if a new
PU type is introduced to an existing system the
architecture across the Trusted Guard is unchanged and
allows for IA consistency across LT2-FTS programs. The use of XML minimizes any performance impact
because a wide array of mature tools is available for
quick XML serialization/deserialization. In fact,
translating XML is faster for Trusted Guards than
translation of other standard protocols. •
Provided a simple path for trusted guard certification. XML is a mature and well-known software
communications mechanism and allows for relatively
easy and fast Information Assurance (IA) accreditation
for Cross-Domain Solutions. Furthermore, even if a new
PU type is introduced to an existing system the
architecture across the Trusted Guard is unchanged and
allows for IA consistency across LT2-FTS programs. The use of XML minimizes any performance impact
because a wide array of mature tools is available for
quick XML serialization/deserialization. In fact,
translating XML is faster for Trusted Guards than
translation of other standard protocols. ACKNOWLEDGEMENTS Live Training Transformation (LT2) Overview, Product line
(2008). Retrieved from http://lt2portal.org The authors wish to thank COL Fred Mullins, Project
Manager Training Devices (PM TRADE), Mr. Wayne
Koenig, Training and Doctrine Command (TRADOC)
Program and Integration Office (TPIO) – Live, and Mr. Robert Parrish, Director Live Engineering Division,
PEO STRI for their vision, contributions and assistance
in the evolution of the LT2 product-line. The authors wish to thank COL Fred Mullins, Project
Manager Training Devices (PM TRADE), Mr. Wayne
Koenig, Training and Doctrine Command (TRADOC)
Program and Integration Office (TPIO) – Live, and Mr. Robert Parrish, Director Live Engineering Division,
PEO STRI for their vision, contributions and assistance
in the evolution of the LT2 product-line. Live Training Transformation (LT2) HCI Style Guide Version
2.1 (2004). Orlando FL: U.S. Army PEO-STRI. Live
Training
Transformation
(LT2)
Product
Line
Management Concept Of Operations, Version 4, (2006). Orlando FL: U.S. Army PEO-STRI. One Tactical Engagement Simulation System (OneTESS)
Capabilities Production Document (CPD). Orlando FL:
U.S. Army PEO-STRI. Video Service-Oriented Architecture (SOA) LT2 has standardized on a service-oriented architecture to
accommodate its video management, transmission, storage,
streaming, viewing and recording requirements. Video
technologies, structures, and even standards are well in place •
Funding consolidation of budget for all LT2-FTS
maximizing each program contribution for the benefits
of all programs 2007 Paper No. 7200 Page 7 of 8 MILCOM 2008 Live Training Transformation (LT2) CTIA Architecture
Change
Process
(ACP). Retrieved
2008
from
http://www.lt2portal.org •
Government and Industry work together to establish
interoperability and technology insertion standards that
foster Government product line evolution and simplify
acquisition processes while allowing industry to
maintain its competitive edge or business model. •
Government and Industry work together to establish
interoperability and technology insertion standards that
foster Government product line evolution and simplify
acquisition processes while allowing industry to
maintain its competitive edge or business model. Live Training Transformation (LT2) Common Component
Change
Process
(C3P). Retrieved
2008
from
http://www.lt2portal.org Collaboration working groups include Government and
industry stakeholders that follow LT2 standard processes
fostering standardized solutions to LT2-FTS identified
product line stovepipe issues and technology gaps. Solutions
support open source delivery and access to existing products
to extend and modernize into new products via LT2 web-
based Portal or CPM repository. Standardization provides the
abilities to create new product lines from our products for
external customers. Collaboration working groups include Government and
industry stakeholders that follow LT2 standard processes
fostering standardized solutions to LT2-FTS identified
product line stovepipe issues and technology gaps. Solutions
support open source delivery and access to existing products
to extend and modernize into new products via LT2 web-
based Portal or CPM repository. Standardization provides the
abilities to create new product lines from our products for
external customers. Live Training Transformation (LT2) Integrated Control
Documents
(ICD). Retrieved
2008
from
http://www.lt2portal.org Live Training Transformation (LT2) Developers Guide,
Version 4, (2006). Orlando FL: U.S. Army PEO-STRI. Live Training Transformation (LT2) DRAFT Consolidated
Product Line Management (CMP) Acquisition Strategy,
V0.1, (2008). Orlando FL: U.S. Army PEO-STRI. REFERENCES Samper, W., Rivera, J. (2007). Applying the Live Training
Transformation (LT2) Software Reuse Strategy to the Home
station
Instrumentation
Training
System. 2007 Samper, W., Rivera, J. (2007). Applying the Live Training
Transformation (LT2) Software Reuse Strategy to the Home
station
Instrumentation
Training
System. 2007
Interservice/Industry Training, Simulation, and Education
Conference (I/ITSEC), Orlando, Florida. Interservice/Industry Training, Simulation, and Education
Conference (I/ITSEC), Orlando, Florida. Dumanoir, P., Rivera, J. (2006). Component-Based Software
Reuse Strategy for Army Live Training Ranges. 2006
Simulation Interoperability Workshop (SIW). LT2 Player Unit Interface Control Document PU ICD version
1.1 , (2008). Orlando FL: U.S. Army PEO-STRI. LT2 Integrated Player Unit Performance Specification
(IPUPS), Version 1.1, (2008). Orlando FL: U.S. Army
PEO-STRI. Combat Training Instrumentation Architecture (CTIA) web
site. Retrieved 2008 from http://www.lt2portal.org Family of Army Systems and Integrated Targetry (FASIT)
Standard and ICDs web site. Retrieved 2008 from
http://www.lt2portal.org Dumanoir, P., Rivera, J. (2005). LT2- A Strategy for Future
Army and Joint Live Training. 2005 Interservice/Industry
Training,
Simulation,
and
Education
Conference
(I/ITSEC), Orlando Florida. 2007 Paper No. 7200 Page 8 of 8
|
https://openalex.org/W2626238502
|
https://rbej.biomedcentral.com/track/pdf/10.1186/s12958-017-0264-3
|
English
| null |
The potential regulatory mechanisms of the gonadotropin-releasing hormone in gonadotropin transcriptions identified with bioinformatics analyses
|
Reproductive biology and endocrinology
| 2,017
|
cc-by
| 7,663
|
© The Author(s). 2017 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and
reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to
the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver
(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. The potential regulatory mechanisms of the
gonadotropin-releasing hormone in
gonadotropin transcriptions identified with
bioinformatics analyses Wei Xiang1, Baoyun Zhang1, Fenglin Lv1, Guangde Feng2, Long Chen1, Fang Yang1, Ke Zhang1, Chunyu Cao1,
Pingqing Wang1* and Mingxing Chu3 * Correspondence: wang_pq@21cn.com
1College of Bioengineering, Chongqing University, Chongqing 400030, China
Full list of author information is available at the end of the article Abstract Background: The regulation of gonadotropin synthesis and release by gonadotropinreleasing hormone (GnRH)
plays an essential role in the neuroendocrine control of reproduction. However, the mechanisms underlying
gonadotropin regulation by GnRH pulse frequency and amplitude are still ambiguous. This study aimed to explore
the molecular mechanisms and biological pathways associated with gonadotropin synthesis by GnRH pulse
frequencies and amplitudes. Methods: Using GSE63251 datasets downloaded from the Gene Expression Omnibus (GEO), differentially expressed
genes (DEGs) were screened by comparing the RNA expression from the GnRH pulse group, the GnRH tonic group
and the control group. Pathway enrichment analyses of DEGs was performed, followed by protein-protein interaction
(PPI) network construction. Furthermore, sub-network modules were constructed by ClusterONE and GO function and
pathways analysed by DAVID. In addition, the relationship between the metabolic pathways and the GnRH pathway
was verified in vitro. Results: In total, 531 common DEGs were identified in GnRH groups, including 290 up-regulated and 241 down-
regulated genes. DEGs predominantly enriched in 16 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways,
including 11 up-regulated pathways (signallingsignallingmetabolic pathways, signallingand GnRH signalling pathway)
and 5 down-regulated pathways (type II diabetes mellitus). Moreover, FBJ osteosarcoma oncogene (FOS) and jun
proto-oncogene (JUN) had higher connectivity degrees in the PPI network. Three modules in the PPI were identified
with ClusterONE. The genes in module 1 were significantly enriched in five pathways, including signallingthe insulin
resistance and GnRH signalling pathway. The genes in modules 2 and 3 were mainly enriched in metabolic pathways
and steroid hormone biosynthesis, respectively. Finally, knockdown leptin receptor (LEPR) and insulin receptor (INSR)
reversed the GnRH-modulated metabolic related-gene expression. Conclusions: The present study revealed the involvement of GnRH in the regulation of gonadotropin biosynthesis and
metabolism in the maintenance of reproduction, achieved by bioinformatics analyses. This, indicates that the GnRH
signalling pathway played a central linkings role in reproductive function and metabolic balance. In addition, the
present study identified the difference response between GnRH pulse and GnRH tone, indicated that abnormal GnRH
pulse and amplitude may cause disease, which may provide an improved understanding of the GnRH pathway and a
new insight for disease diagnosis and treatment. Keywords: Molecular mechanisms, GnRH signalling pathway, Bioinformatics analyses, Metabolic status * Correspondence: wang_pq@21cn.com
1College of Bioengineering, Chongqing University, Chongqing 400030, China
Full list of author information is available at the end of the article © The Author(s). Xiang et al. Reproductive Biology and Endocrinology (2017) 15:46
DOI 10.1186/s12958-017-0264-3 Xiang et al. Reproductive Biology and Endocrinology (2017) 15:46
DOI 10.1186/s12958-017-0264-3 Open Access Background decoding of the pulsatile GnRH signal is essential to elu-
cidate the mechanisms that lead to GnRH pulse fre-
quency dependent differential stimulation of synthesis
and secretion of FSH and LH. g
The hypothalamic-pituitary-gonadal (HPG) axis governs
almost all mammalian reproduction events, from foetal
development, through puberty to sexual maturity [1]. Regulation of the reproductive system is initiated by an
array of external and internal inputs, such as photoperiod,
metabolic products and nutrients, growth factors, stress,
infection and inflammation, as well as many central and
peripheral growth factors and hormones. These inputs are
integrated in the brain and hypothalamus to regulate the
biosynthesis and secretion of gonadotropinreleasing hor-
mone GnRH [2]. GnRH, a hypothalamic decapeptide, is
well known to be4 a crucial factor in normal gonadal de-
velopment and function [3, 4]. Starting at the onset of pu-
berty, GnRH is released in pulses from axon terminals at
the median eminence into the hypothalamic-hypophyseal
portal system and then acts on gonadotrophins in the an-
terior pituitary gland, which express the GnRH receptor. Upon receptor binding, GnRH signalling regulates both
the production and release of gonadotrophins, luteinizing
hormone (LH) and follicle-stimulating hormone (FSH),
which in turn act on their respective receptors in the go-
nads to stimulate steroid hormone release into the blood-
stream [5–7]. LH and FSH then act on the ovary and
testis to stimulate the production of gametes, and steroid
and peptide hormones [8]. These hormones positively and
negatively feedback at the hypothalamus and pituitary
level, thus regulating the reproductive hormone cascade
[8]. Therefore, understanding the molecular mechanisms
involved in the response of gonadotropins to GnRH may
help discover new therapeutic targets for reproductive
disorders and hormone-dependent malignancies. The function of GnRH neurons is controlled and mod-
ulated by a wide range of neuronal systems within the
brain, which transmit feedback signals of sex steroids
and mediate the response to stress, metabolic status and
season [13–15]. Previous studies have recently provided
a better understanding of how GnRH synthesis and
secretion is controlled by modulatory neuronal systems
in the brain. To a large extent, this is due to the recogni-
tion of the roles played by kisspeptin and gonadotropin
inhibitory hormone (GnIH) [16]. A large number of
studies have documented that metabolic status, season,
and stress were able to modulate the function of GnRH
neurons [16], thereby affecting reproduction. Background However,
there is no evidence to indicate how GnRH neurons
modulate the metabolic status, season, and stress, or
how the GnRH signalling pathway interacts with other
related pathways. Therefore, an improved understanding
of the molecular mechanism of GnRH on metabolic sta-
tus is required to develop more specifically therapeutic
approaches for metabolic disease. pp
The availability of online databases and computer
algorithms makes it possible to investigate the complex
mechanisms of pathways. Recently, gene expression pro-
file data associated with the gonadotropin subunit genes
have been studied by many researchers. However, the
detailed mechanism, associated with the gonadotropin
response to GnRH, has not been studied further in a
bioinformatics framework. In the present study, the sig-
nalling mechanisms of gonadotropin responses to GnRH
were explored via a bioinformatics approach using the
microarray
data
GSE63251signalling. Differentially
expressed genes (DEGs) were firstly identified between
cells with and without GnRH, followed by pathway en-
richment analysis and construction of protein–protein
interaction
networks
(PPI). Furthermore,
the
Gene
Ontology (GO) function and Kyoto Encyclopedia of
Genes and Genomes (KEGG) pathway enrichment ana-
lyses of the most remarkable module were performed
using the Database for Annotation, Visualization and
Integrated Discovery (DAVID) software. To our know-
ledge, this is the first report in which GnRH have been
studied in the context of network biology. In addition,
on the basis of networks, we proposed the hypothesis
that GnRH was able to modulate the metabolic state, via
the related membrane receptor, which was verified in
the current study. The findings of this study may provide
a systematic perspective to understand underlying mech-
anisms
and
a
new
insight
of
the
GnRH
role
in
gonadotropin subunit gene transcriptions and metabolic
balance. g
GnRH is released intermittently pulses, and changes in
GnRH pulse frequencies and amplitudes have differential
effects on FSH and LH synthesis and release [9–11]. High GnRH pulse frequencies preferentially stimulate
LH synthesis and secretion, whereas FSH synthesis and
secretion are preferentially stimulated at lower frequen-
cies. Studies using rodent models have demonstrated
that FSHβ gene expression is optimally stimulated by
GnRH pulses every 120 min, whereas LHβ gene expres-
sion is higher at shorter pulse intervals, every 30 min
[10, 12]. The pulsatile manner by which GnRH is
released serves as a primary mechanism by which the
synthesis and secretion of FSH and LH from the gona-
dotropes are controlled. Abstract 2017 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and
reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to
the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver
(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Xiang et al. Reproductive Biology and Endocrinology (2017) 15:46 Xiang et al. Reproductive Biology and Endocrinology (2017) 15:46 Page 2 of 12 Page 2 of 12 Page 2 of 12 Cell culture LβT2 cells, a gift from Dr. Pamela Mellon (University of
California, San Diego) were maintained in high-glucose
DMEM (Mediatech, Inc), supplementedwith 10%foetal
bovine serum (FBS) (Gibico), 100 U of penicillin/mL,
and 100 μg of streptomycin sulphate/mL (Thermo Fisher
Scientific, Inc) in 5% CO2 humidified air at 37 °C [27]. Data preprocessing and differential expression analysis
Background correction and quartile data normalization
was performed on the original array data [19]. The DEGs
(DEG1) of GnRH pulse and the DEGs (DEG2) of GnRH
tonic were respectively analysed with respect to the con-
trol group. Multiple testing was corrected using the
Benjamini–Hochberg [20] procedure to obtain the ad-
justed P-value. Fold changes (FCs) in the expression of
individual genes were calculated and DEGs with P < 0.05
and |log FC| >1 were considered to be significant. DEG1
and DEG2 were then combined and the pooled dataset
was referred to as the overlapping DEGs. Verifying the mechanisms of metabolic pathways in
response to GnRH in vitro Mouse GnRH-10 was synthesized by Shanghai Apeptide
Co Ltd. The interferencethe vectors for LEPR and INSR
were constructed. The sequences that express short hair-
pin RNA (shRNA) targeted to gene mRNA sequences
were cloned in a p-SUPER vector (the Sigma-Aldrich)
(http://www.sigmaaldrich.com/china-mainland.html)
according to manufacturer’s instructions. Affymetrix microarray data The microarray data of GSE63251, contributed by
Lawson et al. [17], were downloaded from the Gene
Expression
Omnibus
(GEO)
(http://www.ncbi.nlm.-
nih.gov/geo/) database at the National Center for
Biotechnology Information (NCBI), which is currently
the largest fully public gene expression resource [18]. The microarray data GSE63251 contains 20 samples,
including two samples of un-stimulated, four samples
of tonic GnRH, and fourteen different GnRH dose
and pulse frequencies. Only six only samples of these
were analysed in the present study, including two
samples of un-stimulated, two samples of perfusion
tonic 100 nM GnRH and two samples of perfusion 8
pulses 100 nM GnRH based on the platform of
GPL81
[MG_U74Av2]
Affymetrix
Murine
Genome
U74A Version 2 Array. Background Changes in the pulse frequency
and amplitude of GnRH release result in differential
FSH and LH synthesis and secretion, contributing to
regulating the female menstrual cycle [9, 11]. However,
the mechanistic basis for the ability of gonadotropes to
distinguish pulse frequency and amplitude are still am-
biguous, and the majority of studies of GnRH signaling
are performed in static culture using tonic treatment of
GnRH. Therefore, a better understanding of the cellular Page 3 of 12 Xiang et al. Reproductive Biology and Endocrinology (2017) 15:46 Page 3 of 12 Functional and pathway enrichment analysis LβT-2 cells were cultured at a in a concentration of
1 × 105 viable cells/ml culture medium, in 12-well plates
at 37 °C and 5% CO2. After 24 h, the medium was
replaced with RPMI only, and then 1.5 μg of empty
pSUPER(with
a
negative
control),
LEPR-shRNA
or
INSR-shRNA vectors were transfected using Lipofecta-
mine 2000. Approximately 8 h after transfection, the
cells were serum starved for 8 h and later stimulated
with GnRH for 24 h, and then total RNA was extracted
using TRIzol reagent. The Kyoto Encyclopedia of Genes and Genomes (KEGG)
[21] knowledge database is a collection of online data-
bases dealing with genomes, enzymatic pathways, and
biological
chemicals. The Database for
Annotation,
Visualization and Integrated Discovery (DAVID) [22],
has been developed as a comprehensive set of functional
annotation tools for relating functional terms with gene
lists using a clustering algorithm. The Gene Ontology
database provides functional unification of large-scale
genomic or transcriptomic data, which mainly includes
three categories, including biological processes (BP), mo-
lecular functions (MF), and cellular components (CC). KEGG pathways of the overlapping DEGs were analysed
with a criterion of P < 0.05 using DAVID 6.7. Reverse transcription quantitative real-time PCR (RT-qPCR)
RNA was extracted using TRIzol reagent (TaKaRa), and
1 μg of total RNA was reverse transcribed into first-
strand cDNA with a mix of oligo-dT and random
primers using the Quantitect Reverse Transcription Kit
(TaKaRa). RT-qPCR was performed on an Applied
Biosystems 7500 Real-Time PCR System using SYBR
green. Standard curves, generated from at least three
dilution series of an abundant sample, were used for the
relative quantification of related genes and β-actin (the
same sample was used to generate standard curves for Methods confidence score of >0.4 were selected to construct the
PPI network using Cytoscape software (version 3.0;
http://cytoscape.org/) [24]. Given that most of the net-
works were scale-free, the hub genes were then selected
with a connectivity degree of >10. Then, a biological net-
work was created between the GnRH signalling pathway
and other related pathways by hub genes. In addition,
based on the degree of connectivity, the enriched func-
tional modules of the PPI network were disclosed using
the ClusterONE Cytoscape plug-in [25, 26]. The signifi-
cance threshold was P < 1 × 10−4. Furthermore, the GO
and KEGG pathway enrichment analyses of the DEGs in
the significant modules were performed using DAVID to
analyse the gene function at the molecular level. PPI network construction and module selection The Search Tool for the Retrieval of Interacting Genes
(STRING, version 9.1, http://www.string-db.org/) [23] is
a web resource and biological database that includes
comprehensively predicted and known interaction infor-
mation. In our study, the overlapping DEGs with a Xiang et al. Reproductive Biology and Endocrinology (2017) 15:46 Xiang et al. Reproductive Biology and Endocrinology (2017) 15:46 Xiang et al. Reproductive Biology and Endocrinology (2017) 15:46 Page 4 of 12 Venn diagram of differentially expressed genes is shown
in Fig. 1. these genes). The expression levels of genes were normal-
ized by dividing by β-actin mRNA expression levels
(primers are shown in Table 1). Dissociation curve analysis
was also performed for each gene at the end of PCR. Each
amplicon generated a single peak. PCR cycling conditions
were as follows: initial denaturation and enzyme activation
at 95 °C for 30 s, followed by 40 cycles of denaturation at
95 °C for 5 s, and annealing at 60 °C for 30 s. these genes). The expression levels of genes were normal-
ized by dividing by β-actin mRNA expression levels
(primers are shown in Table 1). Dissociation curve analysis
was also performed for each gene at the end of PCR. Each
amplicon generated a single peak. PCR cycling conditions
were as follows: initial denaturation and enzyme activation
at 95 °C for 30 s, followed by 40 cycles of denaturation at
95 °C for 5 s, and annealing at 60 °C for 30 s. Pathway enrichment analysis To gain further understanding of the potential functional
roles of the DEGs in GnRH treatment, this study per-
formed a KEGG pathways analysis and set P < 0.05 as
the cut-off value. The results are shown in Table 2. According to the results, up-regulated genes were pri-
marily enriched in pathways associated with MAPK sig-
nalling
pathway,
p53
signalling
pathway,
metabolic
pathways, adipocytokine signalling pathway and GnRH
signalling pathway (Table 2). Down-regulated genes were
primarily enriched in pathways associated with oocyte
meiosis, progesterone-mediated oocyte maturation and
type II diabetes mellitus (Table 2). However, the mecha-
nisms of how gonadotropes are affected by GnRH pulse
frequency and amplitude is not understood. Therefore,
this study was concerned with the differences in gene
expression in GnRH pulse and GnRH tonic, and mainly
focused on the changes that occurred inGnRH pulse, as
these were likely occurred in vivo. Then KEGG pathways
analysis showed that DEGs unique to the GnRH pulse
group were DEGs mainly enriched in dilated cardiomy-
opathy, hypertrophic cardiomyopathy, Alzheimer’s dis-
ease and porphyrin and chlorophyll metabolism, which
were associate with disease (Table 3). Statistical analysis All statistical analysis was performed using Graphpad
Prism software. Data were analysed using unpaired
Student’s t-test or one-way ANOVA and Tukey’s post-
hoc test. Data are presented as means ± SEM of at
least three independent experiments and P < 0.05 was
considered statistically significant. Identification of DEGs The neuropeptide hormone GnRH is a hypothalamic
peptide essential for normal mammalian reproductive
function. Gonadotropin
synthesis
and
release
is
dependent upon pulsatile stimulation by the hypothal-
amic neuropeptide GnRH. However, the role of GnRH
pulse frequency was not examined in depth. This study
investigated the gene profiles that responded to GnRH. Using bioinformatics approaches, a total of 1177 DEGs
between perfusion tonic 100 nM GnRH and control
group with |logFC| > 1 and P < 0.05 were identified,
including 507 up-regulated genes such as jun proto-
oncogene (JUN), and activating transcription factor 4
(ATF 4) and 670 down-regulated DEGs, such as insulin
receptor substrate 1 (IRS 1). In addition, a total of 403
up-regulated (such as activating transcription factor 3)
and 447 down-regulated (such as inositol 1,4,5-trisphos-
phate receptor 2) DEGs were identified between the
GnRH pulses group and control group. The schematic PPI network construction and hub gene identification PPI network construction and hub gene identification
Considering that proteins rarely function alone, it is
necessary to study the protein–protein interactions. Therefore, protein-protein interactions were constructed
in the present study, which helped to understand func-
tions of proteins at the molecule level and explore cell
regulatory mechanisms. Based on the STRING database,
the PPI network of overlapping DEGs contained 334
nodes and 989 edges network were gained with the com-
bined score > 0.4 using Cytoscape (Fig. 2). Proteins with Table 1 The sequences of primers for quantitative fluorescent RT-PCR
Genes
Primers
Length of target fragment, bp
β-Actin
F: 5′- ACTCCTATGTGGGTGACGAGG-3′
137
R: 5′-CACACGCAGCTCATTGTAGAAG-3′
NPY
F:5′- CCAGACAGAGATATGGCAAG-3′
110
R:5′- CATGGAAGGGTCTTCAAGCC-3′
PTPN11
F:5′- TACGGGGTCATGCGTGTTAG-3′
125
R:5′- GAAAGTGGTACTGCCAGACG-3′
GYS1
F:5′- TGGTGGGACCATACACGGA-3′
113
R:5′- TCAGCCAACGCCCAAAATAC-3′
PKLR
F:5′- GCCGCATCTACATTGACGAC-3′
127
R:5′- GCATTTGGCAAGTTCACACC-3′
F, Forward; R, reverse. Table 1 The sequences of primers for quantitative fluorescent RT-PCR Xiang et al. Reproductive Biology and Endocrinology (2017) 15:46 Page 5 of 12 Fig. 1 Schematic Venn diagram of differentially expressed genes. Schematic Venn diagram of differentially expressed genes between the GnRH group
and control group. a Up-regulated DEGs: blue and yellow represent DEGs in the pulse GnRH and tonic GnRH groups respectively, compared with
controls. b Down-regulated DEGs: green and red represent DEGs in the pulse GnRH and tonic GnRH groups, respectively, compared with controls Fig. 1 Schematic Venn diagram of differentially expressed genes. Schematic Venn diagram of differentially expressed genes between the GnRH group
and control group. a Up-regulated DEGs: blue and yellow represent DEGs in the pulse GnRH and tonic GnRH groups respectively, compared with
controls. b Down-regulated DEGs: green and red represent DEGs in the pulse GnRH and tonic GnRH groups, respectively, compared with controls (EGFR), and activating transcription factor 3 (ATF 3). In addition, the result of pathway enrichment showed
that common DEGs were mainly enriched in seven-
teen pathways, but how these signalling pathways
interplay with the GnRH signalling pathway are still
ambiguous. Since hub genes play a crucial role in
many process, we proposed the hypothesis that these
signalling pathways interplayed with GnRH signalling
pathway mainly through hub genes. Based on this, we a very high degree of connectivity are commonly
called “hubs”, and they may play a central regulatory
role in the physiological process. Then, the hub genes
or proteins in the networks with a connectivity degree
of >10 were identified (Table 4, top10). PPI network construction and hub gene identification A total of 53
hub genes were selected from the PPI network, which
included
FBJ
osteosarcoma
oncogene
(FOS),
jun
proto-oncogene (JUN), mitogen-activated protein kin-
ase 1 (MAPK 1), epidermal growth factor receptor Table 2 The enriched KEGG pathway jof DEGs
Up-Regulated
ID
Term
Counts
P-value
Pulse
Constant
Pulse
Constant
mmu04010
MAPK signalling pathway
25
34
3.03E-05
6.49E-09
mmu05412
Arrhythmogenic right ventricular cardiomyopathy (ARVC)
11
9
3.60E-04
0.011371
mmu04520
Adherens junction
11
11
4.01E-04
1.00E-03
mmu04115
p53 signalling pathway
10
10
8.34E-04
1.90E-03
mmu04510
Focal adhesion
17
15
0.002282
0.032455
mmu05200
Pathways in cancer
23
21
0.003327
0.04242
mmu01100
Metabolic pathways
83
111
0.015105
0.008450594
mmu04060
Cytokine-cytokine receptor interaction
19
11
0.033202668
0.048153254
mmu04920
Adipocytokine signalling pathway
7
8
0.034984
0.019396
mmu05020
Prion diseases
5
6
0.037602
0.013348
mmu05416
Viral myocarditis
8
9
0.045733
0.038737
mmu04912
GnRH signalling pathway
11
11
0.046921
0.045341
mmu04012
ErbB signalling pathway
7
9
0.048712
0.025918
Down-Regulated
mmu00280
Valine, leucine and isoleucine degradation
8
0.004814562
6
0.049249252
mmu04114
Oocyte meiosis
13
0.006667
12
0.03641
mmu04914
Progesterone-mediated oocyte maturation
10
0.016259
10
0.031867499
mmu04930
Type II diabetes mellitus
7
0.024595
8
0.012683147
Notes: Count: the number of DEGs
Abbreviations: DEGs differentially expressed genes Table 2 The enriched KEGG pathway jof DEGs Table 2 The enriched KEGG pathway jof DEGs
Up-Regulated Page 6 of 12 Page 6 of 12 Page 6 of 12 Table 3 The enriched KEGG pathway of DEGs which exist in
pulse GnRH group while not exist in tonic GnRH group
ID
Term
Count
P-value
mmu05410
Hypertrophic cardiomyopathy (HCM)
10
0.003372782
mmu05414
Dilated cardiomyopathy
10
0.00618965
mmu04670
Leukocyte transendothelial migration
9
0.049331716
mmu04640
Hematopoietic cell lineage
11
0.005141872
mmu05010
Alzheimer’s disease
15
0.042214081
mmu00860
Porphyrin and chlorophyll
metabolism
5
0.047944857
mmu04020
Calcium signalling pathway
15
0.048934483
Notes: Count: the number of DEGs
Abbreviations: DEGs differentially expressed genes Functional module analysis It is known thatleptin, a
peptide hormone secreted by adipocytes, is encoded by
the “fat gene” [28] which can increase body energy ex-
penditure along with decreased energy intake by sup-
pressing appetite [29], thus playing a crucial role in
metabolism and body growth mediation. Based on this,
we investigated whether GnRH modulates the energy
expenditure and energy intake via LEPR, by knockdown
of LEPR expression. The results showed that the
GnRH-induced NPY (Fig. 6a) and PTPN11 (Fig. 6b)
expression was significantly suppressed when the ex-
pression of LEPR was interfered, genes which are
responsible
for
energy
intake
and
reproduction,
respectively. In addition, the role of GnRH on the insu-
lin signalling pathway was also detected. Interfering
with
LEPR
facilitated
the
GnRH-suppressed
GYS1
expression (Fig. 6c), which promoted glycogenesis. Conversely,
interfering
with
LEPR
significantly
repressed GnRH-induced PKLR expression (Fig. 6d),
which promoted glycolysis. haemoglobin biosynthetic processes, while DEGs in mod-
ule 3 were mainly enriched in cell surface and protein
tyrosine phosphatase activity. haemoglobin biosynthetic processes, while DEGs in mod-
ule 3 were mainly enriched in cell surface and protein
tyrosine phosphatase activity. Functional module analysis y
In order to explore the functions of the networks mod-
ules, we implemented cluster analysis. As shown in Fig. 4,
a total of three significant modules with P < 1 × 10−4were
identified using the ClusterONE plug-in of Cytoscape. Module 1 had 99 nodes and 460 edges, module 2 had 24
nodes and 29 edges and module 3 had 20 nodes and 21
edges. KEGG enrichment in the sub-network is presented
in Table 5. The FoxO signalling pathway, MAPK signalling
pathway, insulin resistance and GnRH signalling pathway
were the predominant pathways enriched by the DEGs of
module 1. The pathways enriched in module 2 were the
glycolysis/gluconeogenesis, biosynthesis of antibiotics and
metabolic pathways. Module 3 was mainly enriched in
steroid hormone biosynthesis and ovarian steroidogenesis
pathways. GO enrichment analysis was also performed
and is presented in Fig. 5. The DEGs in module 1 were
significantly enriched in biological processes, such as the
regulation of transcription from RNA polymerase II pro-
moter, response to drugs. signallingDEGs in module 2
were mainly enriched in GO terms associated with re-
sponse to external stimuli, for example, regulation of eIF2
alpha
phosphorylation
and
negative
regulation
of created a biological network to analyse the interaction
between the signalling pathways through hub genes
using Cytoscape tool (Fig. 3). In fact, the biological
network showed that these signalling pathways were
directly or indirectly linked to each other through
hub genes; for example, GnRH signalling pathways
may regulate INS1 release and type II diabetes melli-
tus via the MKPA signalling pathway. Fig. 2 Protein–protein interaction networks of differentially expressed genes (DEGs). The size of each node is proportional to the degree of
nodes. DEG, differentially expressed gene Fig. 2 Protein–protein interaction networks of differentially expressed genes (DEGs). The size of each node is proportional to the degree of
nodes. DEG, differentially expressed gene Xiang et al. Reproductive Biology and Endocrinology (2017) 15:46 Page 7 of 12 Page 7 of 12 Table 4 The hub proteins in protein–protein interaction
network (top 10)
ID
Degree
ID
Degree
Fos
59
Mapk1
34
Jun
56
Mapk3
34
Myc
40
Atf3
29
Egfr
37
Egr1
27
Cdkn1a
36
Junb
26 Table 4 The hub proteins in protein–protein interaction
network (top 10) this study investigated that the regulatory function of
GnRH on metabolic status. Verifying the mechanisms of metabolic pathways in
response to GnRH in vitro To date, many studies have confirmed that metabolic
status, season, and stress were able to modulate the
function of GnRH neurons [13–15]. However, there was
no evidence that indicated how GnRH neurons modu-
late the metabolic status, season, and stress. Therefore, Fig. 3 Nodes linking the enriched KEGG pathway by hub genes. Hexagon represents each pathway. Round circle indicates hub genes. Pink
hexagons represent up-regulated signalling pathway, and green hexagons represent down-regulated signalling pathway Fig. 3 Nodes linking the enriched KEGG pathway by hub genes. Hexagon represents each pathway. Round circle indicates hub genes. Pink
hexagons represent up-regulated signalling pathway, and green hexagons represent down-regulated signalling pathway Xiang et al. Reproductive Biology and Endocrinology (2017) 15:46 Page 8 of 12 Fig. 4 The significant modules in protein-protein interaction network with ClusterONE (p<10E-5). The node stands for the protein (gene), The
edges/lines indicate interactions between these genes Fig. 4 The significant modules in protein-protein interaction network with ClusterONE (p<10E-5). The node stands for the protein (gene), The
edges/lines indicate interactions between these genes Fig. 4 The significant modules in protein-protein interaction network with ClusterONE (p<10E-5). The node stands for the protein (gene), The
edges/lines indicate interactions between these genes and influence each other. Therefore, it is necessary to in-
vestigate the links between them as well as their shared
regulatory mechanisms. In our study, the mechanism
was analysed by bioinformatics, including DEG screen-
ing, PPI network construction, and module analysis of
the PPI network. Based on these results, a workflow was
described for further exploring the interplay between
pathways involved in the gonadotropin regulatory mech-
anisms, through the construction of a biological network
and extending the GnRH signalling pathway with add-
itional
knowledge
involved
in
other
physiological
processes. Discussion GnRH plays a central role in the control of normal
reproductive function by regulating the synthesis and
release of the gonadotropins LH and FSH [30–32]. The
knownlege of signalling mechanisms involved in gonado-
tropin synthesis have been expanding. However, the
mechanisms
underlying
gonadotropin
regulation
by
GnRH remained to be fully elucidated. Bioinformatics
tools have accelerated the progression of biomedical re-
search, with high-throughput screening being especially
useful. Such tools not only reduce the need for time-
consuming and labour-intensive bench-work, but also
provide critical directions and insights for advanced
studies
[33]. In
this
study,
we
first
systematically
explored the mechanisms of gonadotropin responding to
GnRH by bioinformatics. While standard pathway ana-
lysis investigates each pathway individually, biological
processes are not independent but rather interact with In the present study, the results of the GO and KEGG
pathway
enrichment
analyses
revealed
the
primary
biological processes in which the DEGs were involved,
including focal adhesion, type II diabetes mellitus, glyco-
gen
biosynthesis
and
metabolism,
progesterone-
mediated
oocyte
maturation. Furthermore,
other Xiang et al. Reproductive Biology and Endocrinology (2017) 15:46 Page 9 of 12 Table 5 Enriched pathways of DEGs in sub-network
Category
Term
Count
P-value
Module 1
KEGG_PATHWAY
FoxO signalling pathway
12
2.21E-08
KEGG_PATHWAY
MAPK signalling pathway
15
3.56E-08
KEGG_PATHWAY
GnRH signalling pathway
6
0.04375722
KEGG_PATHWAY
Estrogen signalling pathway
5
0.046988099
KEGG_PATHWAY
Insulin resistance
5
0.045070173
Module 2
KEGG_PATHWAY
Homologous recombination
3
0.001471
KEGG_PATHWAY
DNA replication
3
0.00239
KEGG_PATHWAY
Glycolysis / Gluconeogenesis
3
0.00723
KEGG_PATHWAY
Biosynthesis of antibiotics
4
0.007747
KEGG_PATHWAY
Metabolic pathways
7
0.023793
Module 3
KEGG_PATHWAY
Steroid hormone biosynthesis
2
0.069411
KEGG_PATHWAY
Ovarian steroidogenesis
2
0.073434
Count, numbers of DEGs; GO gen ontology, KEGG Kyoto Encyclopedia of Genes and Genomes Fig. 5 The Gene Ontology (GO) functional of the significant module. The left ordinate of histogram represents the gene counts, and right
ordinate of histogram represents the P-value. BP, biological process; CC, cellular component; MF, molecule function Fig. 5 The Gene Ontology (GO) functional of the significant module. The left ordinate of histogram represents the gene counts, and right
ordinate of histogram represents the P-value. BP, biological process; CC, cellular component; MF, molecule function Fig. 5 The Gene Ontology (GO) functional of the significant module. The left ordinate of histogram represents the gen
ordinate of histogram represents the P-value. BP, biological process; CC, cellular component; MF, molecule function Fig. 5 The Gene Ontology (GO) functional of the significant module. Discussion The left ordinate of histogram represents the gene counts, and righ
ordinate of histogram represents the P-value. BP, biological process; CC, cellular component; MF, molecule function Xiang et al. Reproductive Biology and Endocrinology (2017) 15:46 Page 10 of 12 Fig. 6 Effects of GnRH on metabolic related genes expression by knockdown LEPR or INSR. The LβT2 cells transiently transfected with shRNAs
vector targeting LEPR, INSR or empty vector after GnRH treatment. Then total RNA was extracted for qRT-PCR analysis of the genes expression. A,
NPY related expression; B, PTPN11 related expression; C, GYS1 related expression; D, PKLR related expression Fig. 6 Effects of GnRH on metabolic related genes expression by knockdown LEPR or INSR. The LβT2 cells transiently transfected with shRNAs
vector targeting LEPR, INSR or empty vector after GnRH treatment. Then total RNA was extracted for qRT-PCR analysis of the genes expression. A,
NPY related expression; B, PTPN11 related expression; C, GYS1 related expression; D, PKLR related expression occur [40] eliminating a positive signal to the reproduct-
ive axis. However, there was previously no evidence to
indicate how GnRH neurons modulate the metabolic
status. Based on this, we proposed the hypothesis that
GnRH may modulate the metabolic status through regu-
lating expression of the related genes, such as the mem-
brane receptor LEPR and INSR. In fact, the current
results showed that interfering with LEPR significantly
suppressed GnRH-induced NPY and PTPN11 expres-
sion, which are responsible for energy intake and
reproduction, respectively. Similarly, Interfering with
LEPR facilitated the GnRH-suppressed GYS1 expression,
which promoted the glycogenesis. Conversely, interfer-
ing with LEPR significant repressed GnRH-induced
PKLR expression, which promoted glycolysis. These
results provide a better understanding of the regulation
of GnRH on energy intake and expenditure at the
molecular level, which constitute useful insights into the
relationship between reproduction and metabolic status. pathways associated with GnRH regulation were acti-
vated simultaneously followed by treatment with GnRH,
such as MAPK signalling pathway, cytokine-cytokine
receptor interaction, and most importantly, the GnRH
signalling pathway [34]. Based on the results, it was
revealed that the GnRH signalling pathway interplayed
with other signalling pathways to co-regulate the repro-
ductive function and other biological process. Metabolic dysfunctions are often linked to reproduct-
ive abnormalities [35]. Obesity and conditions with
hyperinsulinemia such as type 2 diabetes mellitus, meta-
bolic syndrome, and polycystic ovary syndrome (PCOS)
are often accompanied by infertility in females [36]. Acknowledgements
f l We are grateful to Dr. Pamela Mellon (University of California, San Diego) for
kindly providing LβT2 cell. Author details
1 1College of Bioengineering, Chongqing University, Chongqing 400030, China. 2Sichuan TQLS Animal Husbandry Science and Technology Co.,LTD, City,
Mianyang, Sichuan 621000, China. 3Key Laboratory of Farm Animal Genetic
Resources and Germplasm Innovation of Ministry of Agriculture, Institute of
Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193,
China. Authors’ contributions WX, BYZ, GDF, LC, FY, KZ and CYC analyzed and interpreted data, drafted or
revised the manuscript, read and approved the final manuscript, and agreed
to be accountable for all aspects of the work. FLL, MXC and PQW made
substantial contributions to conception and design. All authors read and
approved the final manuscript. Funding
h
k This work was supported by the National Natural Science Foundation of
China (Grant no. 31372287), Ministry of Agriculture Transgenic Major Projects
of China (2014ZX0800952B), the Agricultural Science and Technology
Innovation Program of China (ASTIP-IAS13), and National Biological Breeding
Capacity Building and Industrialization Projects (2014–2573). Availability of data and materials The datasets used and/or analyzed during the current study available from
the corresponding author on reasonable request. Discussion In-
sulin, like LH, is a gonadotropin, which increases ste-
roidogenesis and altered follicular maturation in animal
models [37]. In addition to insulin, leptin also served as
a critical factor in the process of fat cell metabolism,
which can convey information about nutritional state to
the reproductive axis [35]. In a normal metabolic state,
leptin increases GnRH activity/secretion (indirectly via
afferent
forebrain
interneurons)
[38]
and
enhances
reproductive function in females [39]. In cases of meta-
bolic dysfunction, like obesity, leptin resistance can Previous studies have demonstrated that different fre-
quencies of pulsatile GnRH result in altered secretion
patterns of FSH and LH. Increasing frequencies results
in preferential secretion of LH, whereas decreasing Page 11 of 12 Xiang et al. Reproductive Biology and Endocrinology (2017) 15:46 Page 11 of 12 Xiang et al. Reproductive Biology and Endocrinology (2017) 15:46 frequencies leads to greater FSH secretion. Because FSH
and LH synthesis and secretion are both modulated by
GnRH through the same G protein-coupled GnRH re-
ceptor, one possible mechanism is that distinct signalling
pathways are activated, depending on GnRH pulse fre-
quency, leading to the differential regulation of FSH and
LH production [41, 42]. However, it remains unknown
how gonadotropes decode the pulsatile GnRH signal to
preferentially synthesize FSH or LH. In the present
study, the difference between GnRH pulse and GnRH
tone were also analysed. As expected, GO analysis and
KEGG pathway enrichment analysis revealed that GnRH
pulse treatment activated the unique pathways. These
pathways are involved in hypertrophic cardiomyopathy,
dilated cardiomyopathy, alzheimer’s disease, as well as
the calcium signalling pathway. These results further in-
dicated that abnormal GnRH pulse and amplitude may
cause disease, which may provide an improved under-
standing of the GnRH pathway and a new insight for
disease diagnosis and treatment. Conclusions In summary, with this study, for the first time, we char-
acterized GnRH to prove an insight into their role in
gonadotropin regulation and metabolic balance using
PPI networks and GO and KEGG enrichment analysis,
which might provide comprehensive bioinformatics ana-
lysis of the mechanisms of GnRH. These results indi-
cated that the GnRH signalling pathway interplayed with
other signalling pathways to co-regulate the reproductive
function and other biological process. In addition, the
difference
between
GnRH
pulse
and
GnRH
tone
revealed that abnormal GnRH pulse and amplitude may
cause disease, which may provide an improved under-
standing of the GnRH pathway and a new insight for
disease
diagnosis
and
treatment. However,
further
experimentation and additional studies are needed to
validate these results. Received: 2 March 2017 Accepted: 8 June 2017 BP: Biological process; CC: Cellular component GnRH: gonadotropinreleasing
hormone; DAVID: Database for annotation, visualization and integrated
discovery; FSH: Follicle-Stimulating hormone; GEO: Gene expression omnibus;
GO: Gene ontology; HPG: Hypothalamic-pituitary-gonadal; KEGG: Kyoto
encyclopedia of genes and genomes; LH: Luteinizing hormone;
MF: Molecular function; PPI: protein-protein interaction Publisher’s Note Springer Nature remains neutral with regard to jurisdictional claims in
published maps and institutional affiliations. References 1. Thackray VG, Mellon PL, Coss D. Hormones in synergy: regulation of the
pituitary Gonadotropin genes. Mol Cell Endocrinol. 2010;314:192–203. pituitary Gonadotropin genes. Mol Cell Endocrinol. 2010;314:192–203. 2. Millar RP, Newton CL. Current and future applications of GnRH, kisspeptin
and neurokinin B analogues. Nat Rev Endocrinol. 2013;9:451–66. p
y
p
g
2. Millar RP, Newton CL. Current and future applications of GnRH, kisspeptin
and neurokinin B analogues. Nat Rev Endocrinol. 2013;9:451–66. 3. Stanislaus D, Pinter JH, Janovick JA, Conn PM. Mechanisms mediating
multiple physiological responses to gonadotropin-releasing hormone. Mol
Cell Endocrinol. 1998;144:1. 4. Alarid ET, Windle JJ, Whyte DB, Mellon PL. Immortalization of pituitary cells
at discrete stages of development by directed oncogenesis in transgenic
mice. Development. 1996;122:3319. 5. Christian CA, Moenter SM. The neurobiology of preovulatory and estradiol-
induced gonadotropin-releasing hormone surges. Endocr Rev. 2010;31:544–77. 6. Herbison AE. Estrogen positive feedback to gonadotropin-releasing hormone
(GnRH) neurons in the rodent: the case for the rostral periventricular area of
the third ventricle (RP3V). Brain Res Rev. 2008;57:277–87. 7. Petersen SL, Ottem EN, Carpenter CD. Direct and indirect regulation of
gonadotropin-releasing hormone neurons by estradiol. Biol Reprod. 2004;69:
1771–8. 8.
Millar RP, Lu ZL, Pawson AJ, Flanagan CA, Morgan K, Maudsley SR.
Gonadotropin-releasing hormone receptors. Endocr Rev. 2004;25:235.
9.
Savoy-Moore RT, Swartz KH. Several GnRH stimulation frequencies
differentially release FSH and LH from isolated, Perfused rat anterior pituitary
cells. Adv Exp Med Biol. 1987;219:641. 7.
Petersen SL, Ottem EN, Carpenter CD. Direct and indirect regulation of
gonadotropin-releasing hormone neurons by estradiol. Biol Reprod. 2004;69:
1771–8. Competing interests
h
h
d
l
h The authors declare that there are no competing interests regarding the
publication of this article. Consent for publication
Not applicable. Consent for publication
Not applicable. Consent for publication
Not applicable. However, there are some limitations in our study. First,
the small amounts of data used in the current study
were downloaded from the GEO database, not generated
by us. Because GEO is a huge data repository, a meta-
analysis of the relevant molecular mechanism of GnRH
may be performed in future studies. Second, the results
were web-based and were not all verified by biological
experiments. Thus, further experimental studies based
on our findings are still needed. Ethics approval and consent to participate Ethics approval and consent to participate
Not applicable. 8.
Millar RP, Lu ZL, Pawson AJ, Flanagan CA, Morgan K, Maudsley SR.
Gonadotropin-releasing hormone receptors. Endocr Rev. 2004;25:235. Abbreviations 8. Millar RP, Lu ZL, Pawson AJ, Flanagan CA, Morgan K, Maudsley SR. Gonadotropin-releasing hormone receptors. Endocr Rev. 2004;25:235. 9. Savoy-Moore RT, Swartz KH. Several GnRH stimulation frequencies
differentially release FSH and LH from isolated, Perfused rat anterior pituitary
cells. Adv Exp Med Biol. 1987;219:641. Page 12 of 12 Page 12 of 12 Xiang et al. Reproductive Biology and Endocrinology (2017) 15:46 10. Thompson IR, Ciccone NA, Zhou Q, Xu S, Khogeer A, Carroll RS, et al. GnRH
pulse frequency control of Fshb Gene expression is mediated via ERK1/2
regulation of ICER. Mol Endocrinol. 2016;30:348–60. 36
Michalakis K, Mintziori G, Kaprara A, Tarlatzis BC, Goulis DG. The complex
interaction between obesity, metabolic syndrome and reproductive axis: a
narrative review. Metab Clin Exp. 2013;62:457–78. 11. Burger LL, Dalkin AC, Aylor KW, Haisenleder DJ, Marshall JC. GnRH pulse
frequency modulation of Gonadotropin subunit Gene transcription in
normal Gonadotropes—assessment by primary transcript assay provides
evidence for roles of GnRH and Follistatin. Endocrinology. 2002;143:3243–9. 37
Wu S, Divall S, Nwaopara A, Radovick S, Wondisford F, Ko C, et al. Obesity-
induced infertility and hyperandrogenism are corrected by deletion of the
insulin receptor in the ovarian theca cell. Diabetes. 2014;63:1270–82. 38
Quennell JH, Mulligan AC, Tups A, Liu XH, Phipps SJ, Kemp CJ, et al. Leptin
indirectly regulates gonadotropin-releasing hormone neuronal function. Endocrinology. 2009;150:2805–12. 12. Dalkin AC, Haisenleder DJ, Ortolano GA, Ellis TR, Marshall JC. The frequency of
gonadotropin-releasing-hormone stimulation differentially regulates
gonadotropin subunit messenger ribonucleic acid expression. 1989;125:917–24. 39
Hausman GJ, Barb CR, Lents CA. Leptin and reproductive function. Biochimie. 2012;94:2075–81. 13
Clarke IJ, Caraty A. Kisspeptin and seasonality of reproduction. Adv Exp Med
Biol. 2013;784:411. 40
Ahima RS, Qi Y, Singhal NS. Adipokines that link obesity and diabetes to the
hypothalamus. Prog Brain Res. 2006;153:155–74. 14
Clarke IJ, Tilbrook AJ. Gonadotropin, neural and hormonal control. Encycl
Neurosci. 2009:959–65. 41
Liu F, Usui I, Evans LG, Austin DA, Mellon PL, Olefsky JM, et al. Involvement
of both G(q/11) and G(s) proteins in gonadotropin-releasing hormone
receptor-mediated signalling in L beta T2 cells. J Biol Chem. 2002;277:
32099–108. 15
Clarke IJ. Interface between metabolic balance and reproduction in ruminants:
focus on the hypothalamus and pituitary. Horm Behav. 2014;66:15–40. 16
Clarke IJ, Arbabi L. New concepts of the central control of reproduction,
integrating influence of stress, metabolic state, and season. Domest Anim
Endocrinol. 2016;56:S165–79. 42
Krsmanovic LZ, Mores N, Navarro CE, Arora KK, Catt KJ. Abbreviations An agonist-induced
switch in G protein coupling of the gonadotropin-releasing hormone
receptor regulates pulsatile neuropeptide secretion. Proc Natl Acad Sci U S
A. 2003;100:2969. 17
Lawson MA, Tsutsumi R, Zhang H, Talukdar I, Butler BK, Santos SJ, et al. Pulse sensitivity of the luteinizing hormone β promoter is determined by a
negative feedback loop involving early growth response-1 and Ngfi-a
binding protein 1 and 2. Mol Endocrinol. 2007;21:1175–91. 18
Barrett T. NCBI GEO: archive for functional genomics data sets—update. Nucleic Acids Res. 2011;39:1005–10. 19
Hulsegge I, Kommadath A, Smits MA. Globaltest and GOEAST: two different
approaches for Gene ontology analysis. BMC Proc. 2009;3:S10. 20
Wang Y, Huang L, Wu S, Jia Y, Yang Y, Luo L, et al. Bioinformatics analyses
of the role of vascular endothelial growth factor in patients with non-small
cell lung cancer. PLoS One. 2014;10:e0139285. 21
Du J, Yuan Z, Ma Z, Song J, Xie X, Chen Y. KEGG-PATH: Kyoto encyclopedia
of genes and genomes-based pathway analysis using a path analysis
model. Mol BioSyst. 2014;10:2441. 22
Huang DW, Sherman BT, Lempicki RA. Systematic and integrative analysis of
large gene lists using DAVID bioinformatics resources. Nat Protoc. 2009;4:44–57 23
Mering CV, Huynen M, Jaeggi D, Schmidt S, Bork P, Snel B. STRING: a
database of predicted functional associations between proteins. Nucleic
Acids Res. 2003;31:258. 24
Saito R, Smoot ME, Ono K, Ruscheinski J, Wang PL, Lotia S, et al. A travel
guide to Cytoscape plugins. Nat Methods. 2012;9:1069. 25
Deng L, Xiong P, Luo Y, Bu X, Qian S, Zhong W. Bioinformatics analysis of
the molecular mechanism of diffuse intrinsic pontine glioma. Oncol Lett. 2016; [cited 2017 Feb 22]; Available from: http://www.spandidos-
publications.com/10.3892/ol.2016.5024 26
Maraziotis IA, Konstantina D, Anastasios B. An in silico method for detecting
overlapping functional modules from composite biological networks. BMC
Syst Biol. 2008;2:93. 26
Maraziotis IA, Konstantina D, Anastasios B. An in silico method for detecting
overlapping functional modules from composite biological networks. BMC
Syst Biol. 2008;2:93. 27
Son YL, Ubuka T, Millar RP, Kanasaki H, Tsutsui K. Gonadotropin-inhibitory
hormone inhibits GnRH-induced Gonadotropin subunit Gene transcriptions
by inhibiting AC/cAMP/PKA-dependent ERK pathway in LβT2 cells. Endocrinology. 2012;153:2332–43. 28
Mann DR, Plant TM. Leptin and pubertal development. Semin Reprod Med. 2002;20:93–102. 29
Luo Q, Li W, Li M, Zhang X, Zhang H. Leptin/leptinR-kisspeptin/kiss1r-GnRH
pathway reacting to regulate puberty onset during negative energy
balance. Life Sci. 2016;153:207–12. 30
Kaiser UB, Conn PM, Chin WW. Xiang et al. Reproductive Biology and Endocrinology (2017) 15:46 Abbreviations Studies of gonadotropin-releasing hormone
(GnRH) action using GnRH receptor-expressing pituitary cell lines. Endocr
Rev. 1997;18:46–70. 30
Kaiser UB, Conn PM, Chin WW. Studies of gonadotropin-releasing hormone
(GnRH) action using GnRH receptor-expressing pituitary cell lines. Endocr
Rev. 1997;18:46–70. • We accept pre-submission inquiries
• Our selector tool helps you to find the most relevant journal
• We provide round the clock customer support
• Convenient online submission
• Thorough peer review
• Inclusion in PubMed and all major indexing services
• Maximum visibility for your research
Submit your manuscript at
www.biomedcentral.com/submit
Submit your next manuscript to BioMed Central
and we will help you at every step: • We accept pre-submission inquiries
• Our selector tool helps you to find the most relevant journal
• We provide round the clock customer support
• Convenient online submission
• Thorough peer review
• Inclusion in PubMed and all major indexing services
• Maximum visibility for your research
Submit your manuscript at
www.biomedcentral.com/submit
Submit your next manuscript to BioMed Central
and we will help you at every step: Submit your next manuscript to BioMed Central
and we will help you at every step: Submit your next manuscript to BioMed Central
and we will help you at every step: Submit your next manuscript to BioMed Central
and we will help you at every step: • We accept pre-submission inquiries
• Our selector tool helps you to find the most relevant journal
• We provide round the clock customer support
• Convenient online submission
• Thorough peer review
• Inclusion in PubMed and all major indexing services
• Maximum visibility for your research
Submit your manuscript at
www.biomedcentral.com/submit
Submit your next manuscript to BioMed Central
and we will help you at every step: 31
Stuart P, Bliss AMN. GnRH signalling, the gonadotrope and endocrine
control of fertility. Front Neuroendocrinol. 2010;31:322. 31
Stuart P, Bliss AMN. GnRH signalling, the gonadotrope and endocrine
control of fertility. Front Neuroendocrinol. 2010;31:322. 32
Ioan A, Ciuleanu T, Guttman T, Ghilezan N. Development of GnRH antagonists
for prostate cancer: new approaches to treatment. Oncologist. 2000;5:162–8. 32
Ioan A, Ciuleanu T, Guttman T, Ghilezan N. Development of GnRH antagonists
for prostate cancer: new approaches to treatment. Oncologist. 2000;5:162–8. 33
Kutmon M, Evelo CT, Coort SL. A network biology workflow to study
transcriptomics data of the diabetic liver. BMC Genomics. 2014;15:971. 34
Ye R-S, Xi Q-Y, Qi Q, Cheng X, Chen T, Li H, et al. Differentially expressed
miRNAs after GnRH treatment and their potential roles in FSH regulation in
porcine anterior pituitary cell. Kim YK, editor. PLoS One. 2013;8:e57156. 34
Ye R-S, Xi Q-Y, Qi Q, Cheng X, Chen T, Li H, et al. Differentially expressed
miRNAs after GnRH treatment and their potential roles in FSH regulation in
porcine anterior pituitary cell. Kim YK, editor. PLoS One. 2013;8:e57156. 35
Klenke U, Taylor-Burds C, Wray S. Metabolic influences on reproduction:
Adiponectin attenuates GnRH neuronal activity in female mice. Endocrinology. 2014;155:1851–63. 35
Klenke U, Taylor-Burds C, Wray S. Metabolic influences on reproduction:
Adiponectin attenuates GnRH neuronal activity in female mice. Endocrinology. 2014;155:1851–63.
|
https://openalex.org/W3206069223
|
https://europepmc.org/articles/pmc8562073?pdf=render
|
English
| null |
Roseburia Abundance Associates With Severity, Evolution and Outcome of Acute Ischemic Stroke
|
Frontiers in cellular and infection microbiology
| 2,021
|
cc-by
| 9,614
|
ORIGINAL RESEARCH
published: 19 October 2021
doi: 10.3389/fcimb.2021.669322 Roseburia Abundance Associates
With Severity, Evolution and
Outcome of Acute Ischemic Stroke
Mengmeng Gu 1, Nihong Chen 1,2, Huanhuan Sun 1, Zhongyuan Li 1, Xiangliang Chen 1*,
Junshan Zhou 1 and Yingdong Zhang 1* 1 Department of Neurology, Nanjing First Hospital, Nanjing Medical University, Nanjing, China, 2 Department of Neurology,
Nanjing Yuhua Hospital, Yuhua Branch of Nanjing First Hospital, Nanjing, China Stroke induces disorder of gut microbiota, however, whether this disorder differs
according to stroke severity and its role in the evolution and outcome of stroke is
currently unknown. Here we explored the composition and structure of fecal
microbiome based on 68 acute ischemic stroke patients presenting with minor
symptoms (admission National Institute of Health Stroke Scale (NIHSS) ≤3) and 67
patients with non-minor stroke (admission NIHSS 4-34) using high-throughput Illumina
sequencing of the 16S rRNA. There was no significant difference in a-diversity indices, but
the principal coordinate analysis of the microbiota indicated clear separation of the two
groups. The significantly enriched butyrate-producing genus Roseburia in the minor
stroke group was negatively correlated with fasting glucose, while the
Erysipelotrichaceae incertae sedis abundant in non-minor stroke patients was positively
correlated with stress hyperglycemia (i.e. fasting glucose/glycated hemoglobin ratio). Moreover, the relative abundance of genus Roseburia was also significantly associated
with the dynamic changes of NIHSS score, as well as short-term and long-term functional
outcomes. Our results suggested that stroke affects microbiota composition in a manner
differentiated by stroke severity, and the enrichment of genus Roseburia may play a
protective role in stroke evolution and outcome. Our findings strengthen the relevance of
specific taxa for stroke severity that might allow targeted therapy in acute ischemic stroke. Edited by:
Marloes Dekker Nitert,
The University of Queensland,
Australia Reviewed by:
Ruoting Xu,
First Affiliated Hospital of Wenzhou
Medical University, China
Katarzyna Winek,
Hebrew University of Jerusalem, Israel Reviewed by:
Ruoting Xu,
First Affiliated Hospital of Wenzhou
Medical University, China
Katarzyna Winek,
Hebrew University of Jerusalem, Israel
*Correspondence:
Yingdong Zhang
zhangyingdong@aliyun.com;
zhangyingdong@njmu.edu.cn
Xiangliang Chen
chenxl@njmu.edu.cn Reviewed by:
Ruoting Xu,
First Affiliated Hospital of Wenzhou
Medical University, China
Katarzyna Winek,
Hebrew University of Jerusalem, Israel *Correspondence:
Yingdong Zhang
zhangyingdong@aliyun.com;
zhangyingdong@njmu.edu.cn
Xiangliang Chen
chenxl@njmu.edu.cn *Correspondence:
Yingdong Zhang
zhangyingdong@aliyun.com;
zhangyingdong@njmu.edu.cn
Xiangliang Chen
chenxl@njmu.edu.cn Specialty section:
This article was submitted to
Microbiome in Health and Disease,
a section of the journal
Frontiers in Cellular and
Infection Microbiology
Received: 18 February 2021
Accepted: 18 May 2021
Published: 19 October 2021 Specialty section:
This article was submitted to
Microbiome in Health and Disease,
a section of the journal
Frontiers in Cellular and
Infection Microbiology Specialty section:
This article was submitted to
Microbiome in Health and Disease,
a section of the journal
Frontiers in Cellular and
Infection Microbiology Keywords: gut microbiota, Roseburia, minor stroke, fasting glucose, prognosis INTRODUCTION Received: 18 February 2021
Accepted: 18 May 2021
Published: 19 October 2021 Ischemic stroke is a major cause of disability and mortality worldwide, most prominently in
adults older than 50 years (Diseases and Injuries, 2020). The severity of neurological deficit is a
crucial predictor of clinical outcomes of stroke (Rost et al., 2016; Wouters et al., 2018). Compared
with patients experiencing a minor stroke, patients with severe stroke have a higher disability rate
(Ferro et al., 2016; Farzadfard et al., 2019) and are more likely to have recurrent vascular events
(Park and Ovbiagele, 2016; Asberg et al., 2018). Severe stroke usually means greater irreversible
neurological deficit after local brain tissue injury. On the other hand, severe stroke may lead to
greater systemic inflammatory response, thus aggravating the symptoms and prognosis of patients Citation: Gu M, Chen N, Sun H, Li Z, Chen X,
Zhou J and Zhang Y (2021)
Roseburia Abundance Associates
With Severity, Evolution and
Outcome of Acute Ischemic Stroke. Front. Cell. Infect. Microbiol. 11:669322. doi: 10.3389/fcimb.2021.669322 October 2021 | Volume 11 | Article 669322 Frontiers in Cellular and Infection Microbiology | www.frontiersin.org Roseburia Abundance and Stroke Severity Gu et al. Gu et al. consecutively recruited from May 2018 to June 2019 with the
following inclusion criteria: 1) aged 50 years or older; 2) local
residents for over 6 months; 3) Magnetic Resonance Imaging
(MRI)-confirmed ischemic stroke in the anterior circulation
within 3 days of symptom onset; and 4) signed written
informed consents. Exclusion criteria included: 1) cerebral
hemorrhagic stroke; 2) a history of chronic inflammatory or
immune diseases (e.g., rheumatoid arthritis, systemic lupus
erythematosus, or inflammatory bowel disease); 3) a history of
severe liver or kidney dysfunction, hematological diseases, and
malignancies; 4) administration of probiotics, antibiotics,
corticosteroids or immunosuppressants within the past 1
months; and 5) insufficient collection of fecal or blood samples. (Okar et al., 2020). Accordingly, a better understanding of the
factors involved after the onset of stroke in view of different
severity and their role in stroke severity trajectory is helpful to
predict stroke prognosis and formulate new prevention and
treatment strategies. In the context of post-stroke modifiable
factors, gut microbiota has emerged as a promising target for
shaping a wide variety of factors that will subsequently affect
stroke severity and disease progression (Prame Kumar and
Wong, 2020). (Okar et al., 2020). Accordingly, a better understanding of the
factors involved after the onset of stroke in view of different
severity and their role in stroke severity trajectory is helpful to
predict stroke prognosis and formulate new prevention and
treatment strategies. In the context of post-stroke modifiable
factors, gut microbiota has emerged as a promising target for
shaping a wide variety of factors that will subsequently affect
stroke severity and disease progression (Prame Kumar and
Wong, 2020). Gut microbiota interacts with the brain following an ischemic
stroke through the bidirectional communication axis known as
the microbiota-gut-brain axis (Cryan et al., 2020). Citation: Previous
studies have shown that gut microbiota not only contributes to
the risk factors of stroke such as hypertension (Silveira-Nunes
et al., 2020), obesity (Thaiss, 2018), atherosclerosis (Liu and Dai,
2020), glucose and lipid metabolism (Morrison and Preston,
2016; Luck et al., 2019; Martin et al., 2019), but might also be a
direct risk factor for stroke (Zeng et al., 2019; Tan et al., 2020). Moreover, the modulation of gut microbiota could have a
positive impact on the progression and outcome of ischemic
stroke (Spychala et al., 2018). Aside from the bottom-up
influences, stroke triggers downstream effects including
increased gastrointestinal permeability and dysbiosis of the gut
microbiota, mainly manifested by the reduction of short-chain
fatty acids (SCFAs)-producing bacteria (Meddings and Swain,
2000; Li et al., 2019; Tan et al., 2021). However, uncertainties
remained about the alterations of gut microbiota in different
stroke severity and the specific bacterial taxa involved in the
dynamic course of neurological deficits due to ischemic stroke. The study was approved by the Ethical Review Board of
Nanjing First Hospital (Nanjing, China). The patients provided
their written informed consent to participate in this study. Baseline Characteristics
and Sample Collection We collected demographic information and medical histories
from all participants by face-to-face interview. The etiology of
ischemic stroke was classified by the Trial of Org 10172 in Acute
Stroke Treatment (TOAST) criteria. Large artery atherosclerosis
refers to the stroke caused by significant (>50%) atherosclerotic
stenosis or occlusion of a major brain artery or branch cortical
artery (Adams et al., 1993). Small artery occlusion refers to a
recent infarction in the territory of one perforating arteriole,
which should be less than 20 mm in its maximum diameter in the
axial plane (Wardlaw et al., 2013). Biochemical parameters
including serum levels of total cholesterol (TC), high-density
lipoprotein cholesterol (HDL), low-density lipoprotein
cholesterol (LDL), fasting glucose, glycated hemoglobin, blood
urea nitrogen (BUN), serum creatinine (Scr) and uric acid (UA)
were collected after overnight fasting within 24 hours of
admission and measured at the hospital central laboratory with
laboratory staff blinded to clinical data. Stress hyperglycemia
(SHG) was also included as a better biomarker of critical illness
than absolute hyperglycemia (Roberts et al., 2015). It was
calculated using the following formula: fasting glucose/glycated
hemoglobin ratio. Stroke severity was assessed by experienced
neurologists (H.S and Z.L) on admission using the National
Institute of Health Stroke Scale (NIHSS) score and retested at 24
hours, 3 days and 7 days. Patients were divided into two groups:
minor stroke, who had admission NIHSS score ≤3 (Wang et al.,
2013), and non-minor stroke with admission NIHSS score > 3. The aims of this study were to (1) investigate differences in
the composition and structure of gut microbiota between
hospitalized ischemic stroke patients with different admission
severity; (2) explore the correlation between stroke-induced
alterations of gut microbiota and biochemical profiles such as
glucose and lipids; (3) evaluate the severity-based discriminative
taxa in relation to the evolution of subsequent stroke severity, the
short-term and long-term functional outcomes. Abbreviations: SCFAs, short-chain fatty acids; MRI, Magnetic Resonance
Imaging; TOAST, the Trial of Org 10172 in Acute Stroke Treatment; TC, total
cholesterol; HDL, high-density lipoprotein cholesterol; LDL, low-density
lipoprotein cholesterol; BUN, blood urea nitrogen; Scr, serum creatinine; UA,
uric acid; SHG, stress hyperglycemia; NIHSS, National Institute of Health Stroke
Scale; mRS, modified Rankin scale; PCR, polymerase chain reaction; OTUs,
operational taxonomic units; RDP, Ribosomal Database Project; PCoA,
principal coordinate analysis; PERMANOVA, Permutational multivariate
analysis of variance; LDA, linear discriminant analysis; LEfSe, linear
discriminant analysis effect size; IQR, interquartile range; SD, standard
deviation; LF, lower fence; UF, upper fence; PSM, propensity score-matched;
CHD, coronary heart disease; F/B ratio, Firmicutes to Bacteroidetes ratio; FDR,
false discovery rate; OR, odds ratio; CI, confidence interval. Study Participants This is a prospective observational cohort study conducted in
Nanjing First Hospital. Patients with acute ischemic stroke were Sterile fecal containers and instructions were distributed to
each study participant on admission. Approximately 2 g of
fresh fecal samples were collected from each participant within
24 hours after admission and immediately (within 1 hour) stored
at -80°C until analysis. Abbreviations: SCFAs, short-chain fatty acids; MRI, Magnetic Resonance
Imaging; TOAST, the Trial of Org 10172 in Acute Stroke Treatment; TC, total
cholesterol; HDL, high-density lipoprotein cholesterol; LDL, low-density
lipoprotein cholesterol; BUN, blood urea nitrogen; Scr, serum creatinine; UA,
uric acid; SHG, stress hyperglycemia; NIHSS, National Institute of Health Stroke
Scale; mRS, modified Rankin scale; PCR, polymerase chain reaction; OTUs,
operational taxonomic units; RDP, Ribosomal Database Project; PCoA,
principal coordinate analysis; PERMANOVA, Permutational multivariate
analysis of variance; LDA, linear discriminant analysis; LEfSe, linear
discriminant analysis effect size; IQR, interquartile range; SD, standard
deviation; LF, lower fence; UF, upper fence; PSM, propensity score-matched;
CHD, coronary heart disease; F/B ratio, Firmicutes to Bacteroidetes ratio; FDR,
false discovery rate; OR, odds ratio; CI, confidence interval. DNA Extraction and High
Throughput Sequencing analysis and linear discriminant analysis (LDA) effect size (LEfSe)
were used to determine the significantly discriminative taxa
between groups (Segata et al., 2011). Bacteria with significant
differences (absolute value of logarithmic LDA score > 2) between
the two groups were plotted on taxonomic bar plots. We also used
BugBase to predict potential microbiome phenotypes, including
aerobic, anaerobic, containing mobile elements, facultatively
anaerobic, biofilm forming, gram-negative, gram-positive,
potentially pathogenic, and stress tolerant (Ward et al., 2017). DNA extraction and sequencing were supported by the Shanghai
Genesky Biotechnology Company (Shanghai, China) not
knowing group assignment. According to the instructions, fecal
genomic DNA was extracted from the fecal samples using the
QIAamp® DNA Stool Mini Kit (Qiagen, Hilden, Germany). The
V3-V4 hypervariable regions of the bacterial 16S rRNA gene
were amplified by polymerase chain reaction (PCR) with the
forward primer (5’-CCTACGGGNGGCWGCAG-3’) and the
reverse primer (5’-GACTACHVGGGTATCTAATCC-3’)
(Meng et al., 2021). Each sample was independently amplified
by three repeated PCR experiments. The PCR products were
checked by agarose gel electrophoresis, and the products from
the same sample were pooled. The pooled PCR product was used
as a template, and the index PCR was performed by using index
primers for adding the Illumina index to the library. The
amplification products were checked using gel electrophoresis
and were purified using the Agencourt AMPure XP Kit
(Beckman Coulter, CA, USA). The purified products were
indexed in the 16S V3-V4 library. The library quality was
assessed on the Qubit@2.0 Fluorometer (Thermo Scientific,
USA) and Agilent Bioanalyzer 2100 systems (USA). High
throughput sequencing was performed on the Illumina Miseq
platform using the 2×250 bp paired-end read protocol. p
y p
g
All statistical analyses were performed with R version 3.4.3 (R
Development Core Team, Vienna, Austria). Continuous variables
were expressed as median [interquartile range (IQR)] or mean ±
standard deviation (SD) and compared with Wilcoxon rank sum
test or student t test when appropriate. Categorical variables were
expressed as number (percentage) and compared by Pearson’s
chi-square test. A box-plot outlier is detected as any observation
that falls below the lower fence (LF) [LF = Q1-1.5*IQR] or above
the upper fence (UF) [UF = Q3+1.5*IQR], where Q1, Q3 and IQR
are the first quartile, third quartile and interquartile range of the
data, respectively (Chervoneva et al., 2006). Functional Outcomes Functional outcomes were quantified using the modified Rankin
scale (mRS) score at 30 days and 1 year through routine
telephone interview (M.G, H.S and Z.L). Poor functional
outcome was defined as mRS score > 2. October 2021 | Volume 11 | Article 669322 Frontiers in Cellular and Infection Microbiology | www.frontiersin.org 2 Roseburia Abundance and Stroke Severity Gu et al. Bioinformatics and Statistical Analysis Bioinformatics and Statistical Analysis
The raw reads were quality filtered and merged with the following
criteria: (1) truncation of the raw reads at any site with an average
quality score < 20, removal of reads contaminated by adapter and
further removal of reads having less than 100 bp by TrimGalore;
(2) the paired end reads were merged to tags by Fast Length
Adjustment of Short reads (FLASH, v1.2.11); (3) removal of reads
with ambiguous bases (N base) and reads with more than 6 bp of
homopolymer by Mothur; (4) removal of reads with low
complexity to obtain clean reads for further bioinformatics
analysis. The remaining unique reads were clustered into
operational taxonomic units (OTUs) by UPARSE with a 97%
similarity cutoff. All OTUs were classified based on Ribosomal
Database Project (RDP) Release 9 by Mothur. Within-individual
(a) diversity (including observed species, Chao 1, ACE, Shannon,
Simpson, and Coverage index) was used to measure the richness
or evenness of taxa within each sample, and was analyzed by
Mothur. Between-individual (b) diversity was provided for
comparison of the taxonomic profiles between microbial
communities. Unweighted UniFrac is a qualitative measure of
the distance between microbial communities by calculating the
fraction of the branch length in a phylogenetic tree (Lozupone
and Knight, 2005). Weighted UniFrac further weights the
branches of a phylogenetic tree based on the abundance of
information, which is a quantitative measure (Chang et al.,
2011). Unweighted and weighted UniFrac principal coordinate
analysis (PCoA) based on OTUs were performed by R version
3.4.3 (Vegan package). Permutational multivariate analysis of
variance (PERMANOVA; Adonis function) was carried out to
examine whether there were statistical differences in bacterial
community composition (b-diversity) between groups. Metastats DNA Extraction and High
Throughput Sequencing The missing values of
TC (1.5%), HDL (1.5%), LDL (1.5%), fasting glucose (3.7%),
glycated hemoglobin ratio (3.7%), and UA (5.9%) were
interpolated with the median. Propensity score-matched (PSM)
analysis was used to obtain matched pairs of samples from the
minor stroke group and the non-minor stroke group. In the PSM
algorithm, the corresponding propensity score of the grouping
variable (minor or non-minor) was calculated for each patient
with a 1:1 nearest-neighbor matching algorithm with a caliper
width of 0.2 of the propensity score, with age, sex, and coronary
heart disease (CHD) as covariates. Spearman’s rank correlation
coefficient was used to explore the correlation of different genera
with biochemical parameters, NIHSS scores obtained at different
timepoints and functional outcomes. We used linear mixed-effects
models with random intercepts and slopes to test whether the
relative abundance of discriminative taxa (e.g., genus Roseburia)
or Firmicutes to Bacteroidetes ratio (F/B ratio) or gram-negative/
gram-positive ratio account for the evolution of NIHSS scores
through the first 7 days of hospitalization. Since the NIHSS score
was highly skewed, the natural logarithm transformation [ln
(NIHSS + 1)] was applied. Grand-mean centering for
continuous covariates with meaningless 0 values (such as age)
was performed. Multivariable logistic regression analyses were
also used to evaluate the associations between the relative
abundance of discriminative taxa and functional outcomes at 30
days and 1 year. The resulting p values were adjusted using the
Benjamini-Hochberg false discovery rate (FDR) correction. Two-
sided p value < 0.05 was considered significant. Potential Microbiome Phenotypes Using BugBase, nine potential phenotypes were predicted
(Table 3). Compared with the non-minor stroke group, the
minor stroke group exhibited significantly lower levels of the
relative abundances of the gram-positive (t test, p = 0.020) and
mobile elements containing (t test, p = 0.010) phenotypes, whilst
higher levels of the relative abundances of the gram-negative
(t test, p = 0.023) and potentially pathogenic (t test, p = 0.008)
phenotypes. There was no significant difference in the ratio of RESULTS Baseline Characteristics of Patients
A total of 732 patients were screened and 135 were recruited
according to the inclusion and exclusion criteria. The flowchart
for the selection is presented in Figure S1. A total of 46 (34.1%)
patients received intravenous thrombolysis. The NIHSS score of
non-minor stroke group ranged from 4 to 34. Patients with October 2021 | Volume 11 | Article 669322 Frontiers in Cellular and Infection Microbiology | www.frontiersin.org 3 Roseburia Abundance and Stroke Severity Gu et al. Proteobacteria. Compared with the minor stroke group, the
relative abundance of phylum Firmicutes was significantly
higher in the non-minor stroke group, while the beneficial
phylum Bacteroidetes was relatively less abundant (Figure 2B). After PSM, the relative abundance of phyla Firmicutes and
Bacteroidetes were still significantly different between the two
groups (Figure S3). Furthermore, the F/B ratio of the non-minor
stroke group was significantly higher than that of the minor
stroke group (Wilcoxon rank sum test, p = 0.001, Table 2). minor stroke (n = 68) were younger than those with non-minor
stroke (n = 67) (t test, p = 0.001). CHD was more common in the
non-minor stroke patients (c2 test, p = 0.008), whereas small
artery occlusion was more common in the minor stroke patients
(c2 test, p < 0.001; Table 1). After PSM, 48 patients in the minor
stroke group were matched to 48 patients in the non-minor
stroke group, and no significant difference was observed in
baseline characteristics between the two groups (Table 1). The Structure of the Gut
Microbiota in Patients LEfSe analysis and LDA score were used to further identify
the microbial species with significant differences between the two
groups from phylum to the genus levels. At the genus level,
Bilophila, Roseburia, and Bacteroides were significantly enriched
in the minor stroke group, while Erysipelotrichaceae incertae
sedis, Enterococcus, and Anaerovorax were more abundant in the
non-minor stroke group (Figure 2C). After PSM, the genus
Bacteroides and Roseburia still increased significantly in patients
with minor stroke (Figure S4). A total of 22,689,893 high-quality reads (mean per subject,
168,073; range, 62,541-992,110) were obtained and analyzed. Of these, 89.0% were clean reads. Reads were clustered into 2,600
OTUs at 97% identity. There was no significant difference in the
a-diversity indices between minor and non-minor stroke,
including the richness (observed species, Chao 1, and ACE)
and diversity (Shannon index, Simpson index, and Coverage
index) of microbial communities, whether before or after PSM
(Wilcoxon rank sum test, p > 0.05) (Figures 1A and S2A). The composition of bacterial communities at the genus level
is shown in Figure 3A. Metastats analysis showed significantly
more abundant SCFAs-producing bacteria (i.e., Roseburia,
Bacteroides and Phascolarctobacterium) in the minor stroke
group than those in the non-minor stroke group. While genus
Dorea, Erysipelotrichaceae incertae sedis, Enterococcus, and
Anaerovorax were more abundant in patients with non-minor
stroke (Figure 3B). The results of PCoA showed that the microbial structure of
patients with minor stroke was different from that of patients
with non-minor stroke in weighted UniFrac distances (Adonis,
p = 0.003; Figure 1B). Adonis using an unweighted UniFrac
distance, however, could not detect community composition
difference between the two groups (Adonis, p = 0.081; Figure
1C). Nevertheless, after PSM, we found significant differences
in microbial communities between groups, which was
consistent with the Adonis results of weighted UniFrac
distance (Figures S2B, C). p
g
PSM, propensity score-matched analysis; SD, standard deviation; BMI, body mass index; TOAST, Trial of Org 10172 in Acute Stroke Treatment; LAA, large artery atherosclerosis; SAO,
small artery occlusion. Bold value: p value with statistical significance.
PSM, propensity score-matched analysis; SD, standard deviation; BMI, body mass index; TOAST, Trial of Org 10172 in Acute Stroke Treatment; LAA, large artery atherosclerosis; SAO,
small artery occlusion. *For 2 patients, BMI was not available. Bold value: p value with statistical significance. Difference in the Microbial Composition
at Different Levels We evaluated the differences in the composition of gut microbial
community. Figure 2A showed that most of the gut bacteria
detected fall into 3 phyla: Firmicutes, Bacteroidetes, and TABLE 1 | Baseline characteristics of patients before PSM and after PSM. Baseline characteristics
Before PSM
After PSM
Minor stroke (n = 68)
Non-minor stroke (n = 67)
P value
Minor stroke (N = 48)
Non-minor stroke (N = 48)
P value
Age, mean ± SD, y
65.0 ± 9.2
71.0 ± 10.4
0.001
66.8 ± 9.1
67.7 ± 9.3
0.625
Male, n (%)
48 (70.6)
43 (64.2)
0.427
36 (75.0)
34 (70.8)
0.646
BMI, mean ± SD, kg/m2*
23.9 ± 3.5
24.7 ± 3.7
0.196
24.2 ± 3.6
25.2 ± 3.4
0.164
Hypertension, n (%)
55 (80.9)
56 (83.6)
0.682
39 (81.3)
39 (81.3)
1.000
Diabetes mellitus, n (%)
27 (39.7)
23 (34.3)
0.518
20 (41.7)
15 (31.3)
0.289
Dyslipidemia, n (%)
42 (61.8)
39 (58.2)
0.673
27 (56.3)
29 (60.4)
0.679
Coronary heart disease, n (%)
2 (2.9)
11 (16.4)
0.008
2 (4.2)
2 (4.2)
1.000
Smoking, n (%)
38 (55.9)
39 (58.2)
0.785
27 (56.3)
31 (64.6)
0.404
Drinking, n (%)
25 (36.8)
29 (43.3)
0.440
17 (35.4)
22 (45.8)
0.299
TOAST, n (%)
<0.001
0.054
LAA
27 (39.7)
26 (38.8)
22 (45.8)
23 (47.9)
SAO
32 (47.1)
13 (19.4)
20 (41.7)
11 (22.9)
Others
9 (13.2)
28 (41.8)
6 (12.5)
14 (29.2)
*For 2 patients, BMI was not available. B ld
l
l
i h
i i
l i
ifi TABLE 1 | Baseline characteristics of patients before PSM and after PSM. October 2021 | Volume 11 | Article 669322 Frontiers in Cellular and Infection Microbiology | www.frontiersin.org Roseburia Abundance and Stroke Severity Gu et al. Gu et al. A
B
C
FIGURE 1 | Comparison of gut microbiota diversity between minor stroke patients and non-minor stroke patients. (A) Within-individual (a) diversity including observed
species, Chao 1 and ACE, Shannon index, Simpson index and coverage index in minor stroke patients (blue) and non-minor stroke patients (orange). Boxes represent
the interquartile ranges, lines inside the boxes denote medians, and circles are outliers. Between-individual (b) diversity, including principal coordinate analysis (PCoA)
based on weighted UniFrac distances (B) and unweighted UniFrac distances (C), was tested by Adonis. The blue circles represent samples of minor stroke patients and
orange circles represent samples of non-minor stroke patients. PCoA, principle coordinate analysis. Difference in the Microbial Composition
at Different Levels A A B B C C FIGURE 1 | Comparison of gut microbiota diversity between minor stroke patients and non-minor stroke patients. (A) Within-individual (a) diversity including observed
species, Chao 1 and ACE, Shannon index, Simpson index and coverage index in minor stroke patients (blue) and non-minor stroke patients (orange). Boxes represent
the interquartile ranges, lines inside the boxes denote medians, and circles are outliers. Between-individual (b) diversity, including principal coordinate analysis (PCoA)
based on weighted UniFrac distances (B) and unweighted UniFrac distances (C), was tested by Adonis. The blue circles represent samples of minor stroke patients and
orange circles represent samples of non-minor stroke patients. PCoA, principle coordinate analysis. gram-negative/gram-positive between the two groups (t test, p =
0.059, Table 3). results showed that the enriched genus Erysipelotrichaceae
incertae sedis in the non-minor stroke group was positively
correlated with the SHG (Spearman correlation, r = 0.21, p <
0.05, Figure 4). While the enriched genus Roseburia in the minor
stroke group was negatively correlated with fasting glucose
(Spearman correlation, r = -0.19, p < 0.05, Figure 4). Likewise,
a similar significantly negative correlation was observed between
the abundance of Roseburia and fasting glucose after PSM
(Spearman correlation, r = -0.23, p < 0.05, Figure S5). Besides,
the butyrate-producing genus Dialister showed a constantly
negative correlation with fasting glucose and SHG, and
Butyricicoccus with BUN. Genus Escherichia/Shigella was Correlation Analysis Between Gut Bacteria
and Biochemical Parameters There were no statistically significant differences in TC, HDL,
LDL, fasting glucose, BUN, Scr and UA levels between the two
groups (Table 2). The levels of SHG and F/B ratio in the non-
minor stroke group were significantly higher than that in the
minor stroke group (Table 2). We performed correlation analyses between the top-50
common genera (Table S1) and biochemical parameters. The October 2021 | Volume 11 | Article 669322 Frontiers in Cellular and Infection Microbiology | www.frontiersin.org 5 Roseburia Abundance and Stroke Severity Gu et al. A
B
C
GURE 2 | Difference of microbial composition between minor stroke patients and non-minor stroke patients. (A) Taxonomic summary of the gut microbiota of minor
roke patients and non-minor stroke patients at the phylum level. (B) Bacteria with significant differences between minor stroke patients (blue) and non-minor stroke patients
range) at the phylum level (Metastats analysis). Boxes represent the interquartile ranges, lines inside the boxes denote medians, and circles are outliers. (C) Significantly
scriminative taxa between the minor stroke patients (blue) and non-minor stroke patients (orange) determined by linear discriminant analysis effect size (LDA effect size). < 0.05, **p < 0.01. A B B C C FIGURE 2 | Difference of microbial composition between minor stroke patients and non-minor stroke patients. (A) Taxonomic summary of the gut microbiota of minor
stroke patients and non-minor stroke patients at the phylum level. (B) Bacteria with significant differences between minor stroke patients (blue) and non-minor stroke patients
(orange) at the phylum level (Metastats analysis). Boxes represent the interquartile ranges, lines inside the boxes denote medians, and circles are outliers. (C) Significantly
discriminative taxa between the minor stroke patients (blue) and non-minor stroke patients (orange) determined by linear discriminant analysis effect size (LDA effect size). *p < 0.05, **p < 0.01. positively correlated with SHG and negatively correlated with
BUN, and Parasutterella was positively correlated with LDL. (Spearman correlation, r = -0.27, p < 0.01), while SCFAs-
producing bacteria Roseburia and Butyricicoccus were
negatively correlated with NIHSS at all times (Spearman
correlation, p < 0.001). Meanwhile, Erysipelotrichaceae incertae
sedis and Enterococcus were significantly positively correlated
with NIHSS scores at all times (Spearman correlation, p < 0.05). Furthermore, SCFAs-producing bacteria Faecalibacterium,
Roseburia and Bacteroides had a negative correlation with
the mRS scores at 30 days and 1 year (Spearman correlation, The Dynamic Association Between Gut
Microbiota and Stroke Severity The Dynamic Association Between Gut
Microbiota and Stroke Severity At 30 days and 1 year, 1 and 5 patients were lost to follow-up,
respectively. As shown in Figure 5, SCFAs-producing bacterium
Bacteroides was negatively correlated with 24-hour (Spearman
correlation, r = -0.26, p < 0.01) and 3-day NIHSS score October 2021 | Volume 11 | Article 669322 Frontiers in Cellular and Infection Microbiology | www.frontiersin.org 6 Roseburia Abundance and Stroke Severity Gu et al. Gu et al. TABLE 2 | Metabolic parameters and microbial composition of patients. Minor stroke (N = 68)
Non-minor stroke (N = 67)
P value
TC, median (IQR), mmol/L
4.32 (3.52-5.07)
4.37 (3.76-4.81)
0.948
HDL, median (IQR), mmol/L
1.08 (0.95-1.20)
1.08 (0.92-1.21)
0.672
LDL, median (IQR), mmol/L
2.48 (1.86-3.17)
2.60 (2.06-3.15)
0.572
Fasting glucose, median (IQR), mmol/L
5.07 (4.41-6.47)
5.39 (4.75-6.90)
0.075
SHG, median (IQR), mmol/L/%
0.87 (0.75-0.96)
0.93 (0.80-1.10)
0.017
BUN, median (IQR), mmol/L
5.62 (4.43-6.66)
5.40 (4.50-6.70)
0.610
Scr, median (IQR), mmoI/L
71.7 (58.4-84.4)
69.9 (59.0-86.0)
0.893
UA, median (IQR), mmoI/L
294.5 (241.8-360.3)
303.0 (219.0-358.5)
0.380
F/B ratio, median (IQR)
1.73 (0.76-5.38)
3.27 (1.48-14.89)
0.001
The number of missing values of TC, HDL, LDL, fasting glucose, SHG and UA were 2, 2, 2, 2, 5, 10 and 8 respectively. Bold value: p value with statistical significance. TC, total cholesterol; IQR, interquartile range; HDL, high-density lipoprotein cholesterol; LDL, low-density lipoprotein cholesterol; SHG, stress hyperglycemia (fasting glucose/glycated
hemoglobin ratio); BUN, blood urea nitrogen; Scr, serum creatinine; UA, uric acid; F/B ratio, Firmicutes to Bacteroidetes ratio. TABLE 2 | Metabolic parameters and microbial composition of patients. Bold value: p value with statistical significance. TC, total cholesterol; IQR, interquartile range; HDL, high-density lipoprotein cholesterol; LDL, low-density lipoprotein cholesterol; SHG, stress hyperglycemia (fasting glucose/glycated
hemoglobin ratio); BUN, blood urea nitrogen; Scr, serum creatinine; UA, uric acid; F/B ratio, Firmicutes to Bacteroidetes ratio. Bold value: p value with statistical significance. TC, total cholesterol; IQR, interquartile range; HDL, high-density lipoprotein cholesterol; LDL, low-density lipoprotein cholesterol; SHG, stress hyperglycemia (fasting glucose/glycated
hemoglobin ratio); BUN, blood urea nitrogen; Scr, serum creatinine; UA, uric acid; F/B ratio, Firmicutes to Bacteroidetes ratio. A
B
FIGURE 3 | Genus-level difference of microbial composition between minor stroke patients and non-minor stroke patients. (A) Taxonomic summary of the gut
microbiota of minor stroke patients and non-minor stroke patients at the genus level. (B) Bacteria with significant differences between minor stroke patients (blue)
and non-minor stroke patients (orange) at the genus level (Metastats analysis). The Dynamic Association Between Gut
Microbiota and Stroke Severity Boxes represent the interquartile ranges, lines inside the boxes denote medians, and
circles are outliers. *p < 0.05, **p < 0.01. B A B A FIGURE 3 | Genus-level difference of microbial composition between minor stroke patients and non-minor stroke patients. (A) Taxonomic summary of the gut
microbiota of minor stroke patients and non-minor stroke patients at the genus level. (B) Bacteria with significant differences between minor stroke patients (blue)
and non-minor stroke patients (orange) at the genus level (Metastats analysis). Boxes represent the interquartile ranges, lines inside the boxes denote medians, and
circles are outliers. *p < 0.05, **p < 0.01. TABLE 3 | Relative abundances of nine potential phenotypes predicted by BugBase in two groups. Phenotypes (proportion)
Minor stroke (N = 68)
Non-minor stroke (N = 67)
P value
Aerobic, mean ± SD
0.01 ± 0.02
0.01 ± 0.02
0.824
Anaerobic, mean ± SD
0.26 ± 0.03
0.24 ± 0.06
0.103
Contains mobile elements, mean ± SD
0.25 ± 0.04
0.27 ± 0.05
0.010
Facultatively anaerobic, mean ± SD
0.02 ± 0.02
0.02 ± 0.02
0.271
Forms biofilms, mean ± SD
0.03 ± 0.02
0.04 ± 0.03
0.234
Gram-negative, mean ± SD
0.09 ± 0.05
0.07 ± 0.05
0.023
Gram-positive, mean ± SD
0.21 ± 0.06
0.23 ± 0.06
0.020
Potentially pathogenic, mean ± SD
0.12 ± 0.05
0.10 ± 0.05
0.008
Stress tolerant, mean ± SD
0.01 ± 0.01
0.01 ± 0.02
0.543
Gram-negative/gram-positive ratio, mean ± SD
0.58 ± 0.63
0.40 ± 0.48
0.059
Bold value: p value with statistical significance. SD, standard deviation. TABLE 3 | Relative abundances of nine potential phenotypes predicted by BugBase in two groups. SD, standard deviation. October 2021 | Volume 11 | Article 669322 Frontiers in Cellular and Infection Microbiology | www.frontiersin.org Roseburia Abundance and Stroke Severity Gu et al. Gu et al. FIGURE 4 | Heatmap of Spearman correlation analysis between gut microbiota and biochemical parameters. *p < 0.05, **p < 0.01. TC, total cholesterol; HDL, high-
density lipoprotein cholesterol; LDL, low-density lipoprotein cholesterol; SHG, stress hyperglycemia; BUN, blood urea nitrogen; Scr, serum creatinine; UA, uric acid. FIGURE 4 | Heatmap of Spearman correlation analysis between gut microbiota and biochemical parameters. *p < 0.05, **p < 0.01. TC, total cholesterol; HDL, high-
density lipoprotein cholesterol; LDL, low-density lipoprotein cholesterol; SHG, stress hyperglycemia; BUN, blood urea nitrogen; Scr, serum creatinine; UA, uric acid. The Dynamic Association Between Gut
Microbiota and Stroke Severity is relatively small (median 0.00407, interquartile range: 0.00045-
0.01324), we multiplied it by 100 to yield a percentage. After
adjusting for sex, age, CHD, stroke etiology and intravenous
thrombolysis, the results of multivariable logistic regression
analyses showed that the relative abundance of genus
Roseburia was negatively associated with poor functional
outcome at 30 days [odds ratio (OR) 0.50, 95% confidence
interval (CI): 0.26-0.96, p = 0.036] and 1 year (OR 0.45, 95%
CI: 0.22-0.92, p = 0.030). However, the relative abundance of
genus Bacteroides was not associated with poor functional
outcomes, whether at 30 days (OR 0.14, 95%CI: 0.01-2.69, p =
0.194) or 1 year (OR 0.05, 95%CI: 0.02-1.15, p = 0.061). In
addition, neither F/B ratio nor gram-negative/gram-positive
ratio were associated with short-term and long-term
functional outcomes. p < 0.05), while a positive correlation was noted for Enterococcus
at both follow-ups (Spearman correlation, p < 0.01), and
Erysipelotrichaceae incertae sedis was correlated with mRS
score only at the 1-year follow-up (Spearman correlation,
r = 0.18, p = 0.038). We chose the significantly discriminative taxa between
groups before and after PSM (genus Bacteroides and
Roseburia) to investigate their associations with short-term and
long-term prognosis. The results of linear mixed-effects model
showed that, the relative abundance of genus Roseburia was
associated with the ln(NIHSS+1) score (estimate = -13.42, p =
0.015). After adjusting for sex, age, CHD, stroke etiology and
intravenous thrombolysis, the relationship was still statistically
significant (estimate = -10.72, p = 0.047). Besides, gram-negative/
gram-positive ratio (estimate = -0.28, p = 0.034) and the relative
abundance of genus Bacteroides (estimate = -1.13, p = 0.010)
were also significantly associated with the ln(NIHSS+1) score. However, F/B ratio (estimate = 0.002, p = 0.167) had no
correlation with the ln(NIHSS+1) score. Frontiers in Cellular and Infection Microbiology | www.frontiersin.org DISCUSSION This prospective cohort of 135 stroke patients showed that
(1) despite the similar a-diversity indices, some gut microbiota
genera, like Roseburia and Erysipelotrichaceae incertae sedis, were
distinct between patients with minor and non-minor stroke; We also used multivariable logistic regression analyses to
investigate the associations between the relative abundance of
genus Roseburia or Bacteroides and 30-day and 1-year functional
outcomes. Considering that the relative abundance of Roseburia October 2021 | Volume 11 | Article 669322 Frontiers in Cellular and Infection Microbiology | www.frontiersin.org Gu et al. Roseburia Abundance and Stroke Severity Gu et al. FIGURE 5 | Heatmap of Spearman correlation analysis between gut microbiota and NIHSS score and functional outcomes. *p < 0.05, **p < 0.01. NIHSS, the
National Institute of Health Stroke Scale; mRS, modified Rankin scale score. FIGURE 5 | Heatmap of Spearman correlation analysis between gut microbiota and NIHSS score and functional outcomes. *p < 0.05, **p < 0.01. NIHSS, the
National Institute of Health Stroke Scale; mRS, modified Rankin scale score. and norank Ruminococcaceae were increased in severe stroke
patients (NIHSS score > 4) (Li et al., 2019). However, similar
results were not confirmed in our study, this might be attributed
to the differences in the definition of minor stroke and the
sample size of the study cohort. In the study of Li et al., there
were 13 patients with severe stroke and 17 patients with mild
stroke. They defined minor stroke as NIHSS score ≤4. In
addition, the time of enrollment may also account for the
discrepancy (Jeon et al., 2020). For instance, they did not
define the onset time of stroke and they collected fecal samples
within 48 hours of admission. (2) Roseburia was negatively correlated with fasting glucose and
Erysipelotrichaceae incertae sedis was positively correlated with
SHG; (3) the relative abundance of genus Roseburia was
significantly associated with the evolution of NIHSS score and
short-term and long-term functional outcomes. g
Our study is one of the few studies thus far demonstrating
the role of gut microbiota in patients with different stroke
severity. Yin et al. found that Bacteroides were depleted
in severe stroke patients compared with mild stroke
patients (NIHSS score ≤4), whereas Escherichia/Shigella
were more abundant in severe stroke patients (NIHSS score >
4) (Yin et al., 2015). DISCUSSION Consistently, we found that patients
with non-minor stroke have lower levels of Bacteroides
than patients with minor stroke, and the genus Escherichia/
Shigella was positively correlated to SHG, a marker of
disease severity (Pan et al., 2017). In another study, Li et al. reported an increase in Enterobacter, Pyramidobacter, and
Lachnospiraceae UCG-001 in mild stroke patients (NIHSS
score ≤4). Meanwhile, genus Ruminococcaceae UCG-002,
Christensenellaceae R-7 group, Ruminococcaceae UCG-005, An important finding of this study is the robust relationship
between genus Roseburia and stroke severity, evolution and
outcome. Genus Roseburia is an important butyrate-producing
bacterium (Boesmans et al., 2018; Kasahara et al., 2018; Seo et al.,
2020). SCFAs, including acetate, propionate, and butyrate, are
important bacterial metabolites and have beneficial effects on
energy metabolism and intestinal barrier integrity (Kasubuchi
et al., 2015). In our study, other SCFAs-producing bacteria such
as Bacteroides and Phascolarctobacterium were also significantly October 2021 | Volume 11 | Article 669322 Frontiers in Cellular and Infection Microbiology | www.frontiersin.org 9 Roseburia Abundance and Stroke Severity Gu et al. more abundant in the minor stroke group than the non-minor
stroke group. Previous studies have confirmed the importance of
butyrate-producing bacteria in regulating blood glucose and
improving insulin sensitivity (Furet et al., 2010). Compared
with patients with normal carbohydrate metabolism, the
concentrations of butyrate-producing bacteria in patients with
type 2 diabetes were lower (Qin et al., 2012; Karlsson et al., 2013). Transplantation of gut microbiota from lean donors to recipients
with metabolic syndrome has been shown to increase levels of
butyrate-producing microbiota and improve insulin sensitivity
(Vrieze et al., 2012). In our study, the genus Roseburia was also
negatively correlated with fasting glucose. In addition, previous
studies have provided strong evidence for butyrate-producing
microbiota against lipid disorders, systemic inflammation, and
atherosclerosis (Gozd-Barszczewska et al., 2017; Kasahara et al.,
2018; van den Munckhof et al., 2018). SCFAs are also considered
as potential mediators of gut microbiota affecting intestinal
immune function (Vinolo et al., 2011). The transplantation of
SCFA-producers could increase the concentration in gut, brain
and plasma, and reduce the neurological deficit and
inflammation after stroke in aged mice (Lee et al., 2020). There
is also evidence that the microbiota-derived SCFAs have also
been shown to modulate poststroke recovery by affecting
systemic and brain resident immune cells (Sadler et al., 2020). Thus, transplantation of fecal bacteria rich in SCFAs and
supplementation of butyrate might be an effective intervention
for poststroke neuroinflammation. DISCUSSION controls (Demirci et al., 2020). Compared with the healthy
control group, the F/B ratio of atherosclerotic patients was
significantly higher (Emoto et al., 2016). At the same time,
studies have shown that a high-fat diet profoundly increases
the F/B ratio, resulting in dysregulation of the gut microbiota
(Marques et al., 2016). Animal experiments have shown that
reducing the F/B ratio of old mice to a level similar to that of
young mice could improve the prognosis of stroke (Spychala
et al., 2018). These might be the reasons why the F/B ratio
predicts the change of stroke severity. However, in this study,
after adjusting for covariates, F/B ratio was no longer associated
with the evolution of stroke severity. This may be attributed to
the older age of patients in the non-minor stroke group. Additional investigations are warranted to further examine the
correlations between the F/B ratio and the stroke severity and its
possible pathophysiological mechanism. The findings of our study should be interpreted with caution
due to the small sample size and study limitations inherent in any
single-center observational analysis. First of all, this is a hospital-
based study making it impractical to obtain fecal samples before
stroke onset, and the absence of healthy controls makes it
different to calibrate the stroke-induced change in the
composition of gut microbiota. As such, further studies are
needed to explore the effects of the stroke itself on individual
gut microbiota. Secondly, we collected the fecal microbiota and
measured the biochemical parameters at a single time point,
therefore, we could not observe the dynamic changes of the
microbiota to test the real-time interaction with these parameters
and stroke severity. Thirdly, although we included relatively
homogenous participants (i.e. local residents) and excluded
those who had used probiotics, antibiotics, corticosteroids, or
immunosuppressants in the past month, dietary information and
other drugs that may affect gut microbiota were not considered. Another limitation is the lack of characterization of the
metabolomics profiles and SCFAs concentrations in the fecal
and blood samples. Finally, the observed results might be
overstated without the functional correlation or intervention
experiments in human or animal models. However, it provides
the rationale for large-scale and well-designed studies to explore
the prognostic and therapeutic potentials of the genus Roseburia
in stroke patients. In our study, Erysipelotrichaceae incertae sedis was more
abundant in patients with non-minor stroke, and was
positively correlated with SHG and NIHSS scores. DISCUSSION The role of
family Erysipelotrichaceae in metabolic disorders might account
for its potential detrimental effect on stroke severity. Early
evidence suggested that species belonging to Erysipelotrichaceae
would proliferate in diet-induced obese animals (Turnbaugh
et al., 2008). The subsequent research observed an increase of
Erysipelotrichaceae in mice on high-fat or western diet (Fleissner
et al., 2010). Nutrition studies further support the effect of dietary
fat on the abundance of Erysipelotrichaceae (Kaakoush, 2015). Future investigations are needed on the metabolic interactions
with Erysipelotrichaceae under disease states such as a stroke. Considering the high disability rate of stroke, it is important
to predict the change of stroke severity. As a result of this
paper, some bacteria such as Roseburia, Enterococcus, and
Erysipelotrichaceae incertae sedis were significantly correlated
with the NIHSS score. Besides, the F/B ratio of non-minor
stroke patients was higher than that of minor stroke
patients, which was consistent with previous reports (Tan
et al., 2021). Several previous reports demonstrated that the F/
B ratio was associated with obesity, hypertension, diabetes, and
atherosclerosis, which are risk factors for stroke. Obese
individuals had a significantly higher level of Firmicutes and
lower level of Bacteroidetes compared with lean controls (Ley
et al., 2006; Koliada et al., 2017). An increased F/B ratio was
observed both in animal and human hypertension (Yang et al.,
2015). Demirci et al. reported that the F/B ratio in patients with
type 1 diabetes mellitus was significantly lower than in healthy In summary, our study suggests that acute ischemic stroke
patients with different severity have different gut microbiological
characteristics. Moreover, the levels of some gut microbiota were
related to glucose and lipid metabolism. The relative abundance
of genus Roseburia could be a predictor of evolution in stroke
severity and functional outcomes. REFERENCES Furet J. P., Kong L. C., Tap J., Poitou C., Basdevant A., Bouillot J. L., et al. (2010). Differential Adaptation of Human Gut Microbiota to Bariatric Surgery-
Induced Weight Loss: Links With Metabolic and Low-Grade Inflammation
Markers. Diabetes 59, 3049–3057. doi: 10.2337/db10-0253 Adams H. P.Jr., Bendixen B. H., Kappelle L. J., Biller J., Love B. B., Gordon D. L.,
et al. (1993). Classification of Subtype of Acute Ischemic Stroke. Definitions for
Use in a Multicenter Clinical Trial. TOAST. Trial of Org 10172 in Acute Stroke
Treatment. Stroke 24, 35–41. doi: 10.1161/01.str.24.1.35 Markers. Diabetes 59, 3049–3057. doi: 10.2337/db10-0253 Gozd-Barszczewska A., Koziol-Montewka M., Barszczewski P., Mlodzinska A.,
and Huminska K. (2017). Gut Microbiome as a Biomarker of Cardiometabolic
Di
d
A
A i
E
i
M d 24 416 422 d i 10 26444/
/75456 Asberg S., Farahmand B., Hasvold P., Johansson S., and Appelros P. (2018). Non-
Cardioembolic TIA and Ischemic Stroke: Implications of Severity. Acta Neurol. Scand. 138, 369–376. doi: 10.1111/ane.12974 Jeon J., Lourenco J., Kaiser E. E., Waters E. S., Scheulin K. M., Fang X., et al. (2020). Dynamic Changes in the Gut Microbiome At the Acute Stage of Ischemic
Stroke in a Pig Model. Front. Neurosci. 14, 587986. doi: 10.3389/
fnins.2020.587986 Jeon J., Lourenco J., Kaiser E. E., Waters E. S., Scheulin K. M., Fang X., et al. (2020). Dynamic Changes in the Gut Microbiome At the Acute Stage of Ischemic Boesmans L., Valles-Colomer M., Wang J., Eeckhaut V., Falony G., Ducatelle R.,
et al. (2018). Butyrate Producers as Potential Next-Generation Probiotics:
Safety Assessment of the Administration of Butyricicoccus Pullicaecorum to
Healthy Volunteers. mSystems 3, e00094–18. doi: 10.1128/mSystems.00094-18 Kaakoush N. O. (2015). Insights Into the Role of Erysipelotrichaceae in the Human
Host. Front. Cell Infect. Microbiol. 5, 84. doi: 10.3389/fcimb.2015.00084 Chang Q., Luan Y., and Sun F. (2011). Variance Adjusted Weighted UniFrac: A
Powerful Beta Diversity Measure for Comparing Communities Based on
Phylogeny. BMC Bioinformatics 12, 118. doi: 10.1186/1471-2105-12-118 Karlsson F. H., Tremaroli V., Nookaew I., Bergstrom G., Behre C. J., Fagerberg B.,
et al. (2013). Gut Metagenome in European Women With Normal, Impaired
and Diabetic Glucose Control. Nature 498, 99–103. doi: 10.1038/nature12198 Chervoneva I., Hyslop T., Iglewicz B., Johns L., Wolfe H. R., Schulz S., et al. (2006). Statistical Algorithm for Assuring Similar Efficiency in Standards and Samples
for Absolute Quantification by Real-Time Reverse Transcription Polymerase
Chain Reaction. Anal. Biochem. 348, 198–208. REFERENCES doi: 10.1016/j.ab.2005.10.042 Kasahara K., Krautkramer K. A., Org E., Romano K. A., Kerby R. L., Vivas E. I.,
et al. (2018). Interactions Between Roseburia Intestinalis and Diet Modulate
Atherogenesis in a Murine Model. Nat. Microbiol. 3, 1461–1471. doi: 10.1038/
s41564-018-0272-x Cryan J. F., O’Riordan K. J., Sandhu K., Peterson V., and Dinan T. G. (2020). The
Gut Microbiome in Neurological Disorders. Lancet Neurol. 19, 179–194. doi: 10.1016/S1474-4422(19)30356-4 Kasubuchi M., Hasegawa S., Hiramatsu T., Ichimura A., and Kimura I. (2015). Dietary Gut Microbial Metabolites, Short-Chain Fatty Acids, and Host
Metabolic Regulation. Nutrients 7, 2839–2849. doi: 10.3390/nu7042839 Demirci M., Bahar Tokman H., Taner Z., Keskin F. E., Cagatay P., Ozturk Bakar
Y., et al. (2020). Bacteroidetes and Firmicutes Levels in Gut Microbiota and
Effects of Hosts TLR2/TLR4 Gene Expression Levels in Adult Type 1 Diabetes
Patients in Istanbul, Turkey. J. Diabetes Complicat. 34, 107449. doi: 10.1016/
j.jdiacomp.2019.107449 Koliada A., Syzenko G., Moseiko V., Budovska L., Puchkov K., Perederiy V., et al. (2017). Association Between Body Mass Index and Firmicutes/Bacteroidetes
Ratio in an Adult Ukrainian Population. BMC Microbiol. 17, 120. doi: 10.1186/
s12866-017-1027-1 Lee J., d’Aigle J., Atadja L., Quaicoe V., Honarpisheh P., Ganesh B. P., et al. (2020). Gut Microbiota-Derived Short-Chain Fatty Acids Promote Poststroke Recovery
in Aged Mice. Circ. Res. 127, 453–465. doi: 10.1161/CIRCRESAHA.119.316448 Diseases G. B. D., and Injuries C. (2020). Global Burden of 369 Diseases and
Injuries in 204 Countries and Territories 1990-2019: A Systematic Analysis for
the Global Burden of Disease Study 2019. Lancet 396, 1204–1222. doi: 10.1016/
S0140-6736(20)30925-9 Ley R. E., Turnbaugh P. J., Klein S., and Gordon J. I. (2006). Microbial Ecology:
Human Gut Microbes Associated With Obesity. Nature 444, 1022–1023. doi: 10.1038/4441022a Emoto T., Yamashita T., Sasaki N., Hirota Y., Hayashi T., So A., et al. (2016). Analysis of Gut Microbiota in Coronary Artery Disease Patients: A Possible
Link Between Gut Microbiota and Coronary Artery Disease. J. Atheroscler. Thromb. 23, 908–921. doi: 10.5551/jat.32672 Liu Y., and Dai M. (2020). Trimethylamine N-Oxide Generated by the Gut Microbiota
Is Associated With Vascular Inflammation: New Insights Into Atherosclerosis. Mediators Inflamm. 2020, 4634172. doi: 10.1155/2020/4634172 Farzadfard M. T., Sheikh Andalibi M. S., Thrift A. G., Morovatdar N., Stranges S.,
Amiri A., et al. (2019). Long-Term Disability After Stroke in Iran: Evidence
From the Mashhad Stroke Incidence Study. Int. J. Stroke 14, 44–47. doi: 10.1177/1747493018789839 Li N., Wang X., Sun C., Wu X., Lu M., Si Y., et al. (2019). AUTHOR CONTRIBUTIONS The Supplementary Material for this article can be found online
at: https://www.frontiersin.org/articles/10.3389/fcimb.2021. 669322/full#supplementary-material MG, XC and YZ conceived the study. MG and NC analyzed the
data. MG, HS and ZL interpreted the data. MG wrote the MG, XC and YZ conceived the study. MG and NC analyzed the
data. MG, HS and ZL interpreted the data. MG wrote the FUNDING The study was approved by the Ethical Review Board of Nanjing
First Hospital (Nanjing, China). The patients/participants
provided their written informed consent to participate in
this study. This study was supported by National Natural Science
Foundation of China (81701064) and Natural Science
Foundation of Jiangsu Province (BK20201117). DATA AVAILABILITY STATEMENT The datasets presented in this study can be found in online
repositories. The names of the repository/repositories and
accession number(s) can be found below: FigShare repository
(https://figshare.com/), accession number Genome Sequence
Archive repository (DOI:10.6084/m9.figshare.13096223); October 2021 | Volume 11 | Article 669322 Frontiers in Cellular and Infection Microbiology | www.frontiersin.org 10 Roseburia Abundance and Stroke Severity Gu et al. (https://bigd.big.ac.cn/gsa/browse/CRA004134) (Wang et al.,
2017), accession number CRA004134. manuscript. XC and JZ revised the manuscript. All authors
contributed to the article and approved the submitted version. REFERENCES doi: 10.1111/ene.12837 Ward T., Larson J., Meulemans J., Hillmann B., Lynch J., Sidiropoulos D., et al. (2017) BugBase Predicts Organism Level Microbiome Phenotypes. bioRxiv
[Preprint]. doi: 10 .1101/133462. Prame Kumar K., and Wong C. H. (2020). Imbalance in the Force: The Dark Side
of the Microbiota on Stroke Risk and Progression. Curr. Opin. Neurobiol. 62,
10–16. doi: 10.1016/j.conb.2019.10.002 Wardlaw J. M., Smith E. E., Biessels G. J., Cordonnier C., Fazekas F., Frayne R.,
et al. (2013). Neuroimaging Standards for Research Into Small Vessel Disease
and Its Contribution to Ageing and Neurodegeneration. Lancet Neurol. 12,
822–838. doi: 10.1016/S1474-4422(13)70124-8 Qin J., Li Y., Cai Z., Li S., Zhu J., Zhang F., et al. (2012). A Metagenome-Wide
Association Study of Gut Microbiota in Type 2 Diabetes. Nature 490, 55–60. doi: 10.1038/nature11450 Wouters A., Nysten C., Thijs V., and Lemmens R. (2018). Prediction of Outcome in
Patients With Acute Ischemic Stroke Based on Initial Severity and Improvement
in the First 24 H. Front. Neurol. 9, 308. doi: 10.3389/fneur.2018.00308 Roberts G. W., Quinn S. J., Valentine N., Alhawassi T., O’Dea H., Stranks S. N.,
et al. (2015). Relative Hyperglycemia, a Marker of Critical Illness: Introducing
the Stress Hyperglycemia Ratio. J. Clin. Endocrinol. Metab. 100, 4490–4497. doi: 10.1210/jc.2015-2660 Yang T., Santisteban M. M., Rodriguez V., Li E., Ahmari N., Carvajal J. M., et al. (2015). Gut Dysbiosis Is Linked to Hypertension. Hypertension 65, 1331–1340. doi: 10.1161/HYPERTENSIONAHA.115.05315 Rost N. S., Bottle A., Lee J. M., Randall M., Middleton S., Shaw L., et al. (2016). Stroke Severity is a Crucial Predictor of Outcome: An International Prospective
Validation Study. J. Am. Heart Assoc. 5, e002433. doi: 10.1161/
JAHA.115.002433 Yin J., Liao S. X., He Y., Wang S., Xia G. H., Liu F. T., et al. (2015). Dysbiosis of Gut
Microbiota With Reduced Trimethylamine-N-Oxide Level in Patients With
Large-Artery Atherosclerotic Stroke or Transient Ischemic Attack. J. Am. Heart Assoc. 4, e00269. doi: 10.1161/JAHA.115.002699 Zeng X., Gao X., Peng Y., Wu Q., Zhu J., Tan C., et al. (2019). Higher Risk of
Stroke Is Correlated With Increased Opportunistic Pathogen Load and
Reduced Levels of Butyrate-Producing Bacteria in the Gut. Front. Cell Infect. Microbiol. 9, 4. doi: 10.3389/fcimb.2019.00004 Sadler R., Cramer J. V., Heindl S., Kostidis S., Betz D., Zuurbier K. R., et al. (2020). Short-Chain Fatty Acids Improve Poststroke Recovery Via Immunological
Mechanisms. J. Neurosci. 40, 1162–1173. doi: 10.1523/JNEUROSCI.1359-19.2019 Segata N., Izard J., Waldron L., Gevers D., Miropolsky L., Garrett W. REFERENCES Change of Intestinal
Microbiota in Cerebral Ischemic Stroke Patients. BMC Microbiol. 19, 191. doi: 10.1186/s12866-019-1552-1 Lozupone C., and Knight R. (2005). UniFrac: A New Phylogenetic Method for
Comparing Microbial Communities. Appl. Environ. Microbiol. 71, 8228–8235. doi: 10.1128/AEM.71.12.8228-8235.2005 Ferro J. M., Caeiro L., and Figueira M. L. (2016). Neuropsychiatric Sequelae of
Stroke. Nat. Rev. Neurol. 12, 269–280. doi: 10.1038/nrneurol.2016.46 Fleissner C. K., Huebel N., Abd El-Bary M. M., Loh G., Klaus S., and Blaut M. (2010). Absence of Intestinal Microbiota Does Not Protect Mice From Diet-
Induced Obesity. Br. J. Nutr. 104, 919–929. doi: 10.1017/S0007114510001303 Luck H., Khan S., Kim J. H., Copeland J. K., Revelo X. S., Tsai S., et al. (2019). Gut-
associated IgA(+) Immune Cells Regulate Obesity-Related Insulin Resistance. Nat. Commun. 10, 3650. doi: 10.1038/s41467-019-11370-y October 2021 | Volume 11 | Article 669322 Frontiers in Cellular and Infection Microbiology | www.frontiersin.org 11 Roseburia Abundance and Stroke Severity Gu et al. Gu et al. Marques C., Meireles M., Norberto S., Leite J., Freitas J., Pestana D., et al. (2016). High-Fat Diet-Induced Obesity Rat Model: A Comparison Between Wistar and
Sprague-Dawley Rat. Adipocyte 5, 11–21. doi: 10.1080/21623945.2015.1061723 Tan C., Wu Q., Wang H., Gao X., Xu R., Cui Z., et al. (2021). Dysbiosis of Gut
Microbiota and Short-Chain Fatty Acids in Acute Ischemic Stroke and the
Subsequent Risk for Poor Functional Outcomes. JPEN J. Parenter. Enteral Nutr
45, 518–529. doi: 10.1002/jpen.1861 Martin A. M., Yabut J. M., Choo J. M., Page A. J., Sun E. W., Jessup C. F., et al. (2019). The Gut Microbiome Regulates Host Glucose Homeostasis Via
Peripheral Serotonin. Proc. Natl. Acad. Sci. U. S. A. 116, 19802–19804. doi: 10.1073/pnas.1909311116 Thaiss C. A. (2018). Microbiome Dynamics in Obesity. Science 362, 903–904. doi: 10.1126/science.aav6870 Turnbaugh P. J., Backhed F., Fulton L., and Gordon J. I. (2008). Diet-Induced Obesity is
Linked to Marked But Reversible Alterations in the Mouse Distal Gut Microbiome. Cell Host Microbe 3 213 223 doi: 10 1016/j chom 2008 02 015 Turnbaugh P. J., Backhed F., Fulton L., and Gordon J. I. (2008). Diet-Induced Obesity is
Linked to Marked But Reversible Alterations in the Mouse Distal Gut Microbiome. g
J ,
,
,
J
(
)
y
Linked to Marked But Reversible Alterations in the Mouse Distal Gut Microbiome. Cell Host Microbe 3, 213–223. doi: 10.1016/j.chom.2008.02.015 Meddings J. B., and Swain M. G. (2000). REFERENCES Environmental Stress-Induced
Gastrointestinal Permeability Is Mediated by Endogenous Glucocorticoids in
the Rat. Gastroenterology 119, 1019–1028. doi: 10.1053/gast.2000.18152 Cell Host Microbe 3, 213–223. doi: 10.1016/j.chom.2008.02.015 van den Munckhof I. C. L., Kurilshikov A., Ter Horst R., Riksen N. P., Joosten L. A. van den Munckhof I. C. L., Kurilshikov A., Ter Horst R., Riksen N. P., Joosten L. A. B., Zhernakova A., et al. (2018). Role of Gut Microbiota in Chronic Low-Grade
Inflammation as Potential Driver for Atherosclerotic Cardiovascular Disease:
A Systematic Review of Human Studies. Obes. Rev. 19, 1719–1734. doi: 10.1111/obr.12750 Meng Q., Ma M., Zhang W., Bi Y., Cheng P., Yu X., et al. (2021). The Gut
Microbiota During the Progression of Atherosclerosis in the Perimenopausal
Period Shows Specific Compositional Changes and Significant Correlations
With Circulating Lipid Metabolites. Gut Microbes 13, 1–27. doi: 10.1080/
19490976.2021.1880220 Vinolo M. A., Rodrigues H. G., Nachbar R. T., and Curi R. (2011). Regulation of
Inflammation by Short Chain Fatty Acids. Nutrients 3, 858–876. doi: 10.3390/
nu3100858 Morrison D. J., and Preston T. (2016). Formation of Short Chain Fatty Acids by
the Gut Microbiota and Their Impact on Human Metabolism. Gut Microbes 7,
189–200. doi: 10.1080/19490976.2015.1134082 Vrieze A., Van Nood E., Holleman F., Salojarvi J., Kootte R. S., Bartelsman J. F.,
et al. (2012). Transfer of Intestinal Microbiota From Lean Donors Increases
Insulin Sensitivity in Individuals With Metabolic Syndrome. Gastroenterology
143, 913–916 e917. doi: 10.1053/j.gastro.2012.06.031 Okar S. V., Topcuoglu M. A., Yemisci M., Cakir Aktas C., Oguz K. K., and Arsava
E. M. (2020). Post-Stroke Inflammatory Response Is Linked to Volume Loss in
the Contralateral Hemisphere. J. Neuroimmunol. 344, 577247. doi: 10.1016/
j.jneuroim.2020.577247 Wang Y., Song F., Zhu J., Zhang S., Yang Y., Chen T., et al. (2017). GSA: Genome
Sequence Archive. Genomics Proteomics Bioinf. 15, 14–18. doi: 10.1016/
j.gpb.2017.01.001 Pan Y., Cai X., Jing J., Meng X., Li H., Wang Y., et al. (2017). Stress Hyperglycemia
and Prognosis of Minor Ischemic Stroke and Transient Ischemic Attack: The
CHANCE Study (Clopidogrel in High-Risk Patients With Acute Nondisabling
Cerebrovascular Events). Stroke 48, 3006–3011. doi: 10.1161/
STROKEAHA.117.019081 Wang Y., Wang Y., Zhao X., Liu L., Wang D., Wang C., et al. (2013). Clopidogrel
With Aspirin in Acute Minor Stroke or Transient Ischemic Attack. N. Engl. J. Med. 369, 11–19. doi: 10.1056/NEJMoa1215340 Park J. H., and Ovbiagele B. (2016). Relationship of Functional Disability After a
Recent Stroke With Recurrent Stroke Risk. Eur. J. Neurol. 23, 361–367. REFERENCES S., et al. (2011). Metagenomic Biomarker Discovery and Explanation. Genome Biol. 12,
R60. doi: 10.1186/gb-2011-12-6-r60 Conflict of Interest: The authors declare that the research was conducted in the
absence of any commercial or financial relationships that could be construed as a
potential conflict of interest. Seo B., Jeon K., Moon S., Lee K., Kim W. K., Jeong H., et al. (2020). Roseburia Spp. Abundance Associates With Alcohol Consumption in Humans and Its
Administration Ameliorates Alcoholic Fatty Liver in Mice. Cell Host Microbe
27, 25–40 e26. doi: 10.1016/j.chom.2019.11.001 Publisher’s Note: All claims expressed in this article are solely those of the authors
and do not necessarily represent those of their affiliated organizations, or those of
the publisher, the editors and the reviewers. Any product that may be evaluated in
this article, or claim that may be made by its manufacturer, is not guaranteed or
endorsed by the publisher. Silveira-Nunes G., Durso D. F., Alves de Oliveira L. R.Jr, Cunha E. H. M., Maioli T. U., Vieira A. T., et al. (2020). Hypertension Is Associated With Intestinal
Microbiota Dysbiosis and Inflammation in a Brazilian Population. Front. Pharmacol. 11, 258. doi: 10.3389/fphar.2020.00258 Spychala M. S., Venna V. R., Jandzinski M., Doran S. J., Durgan D. J., Ganesh B. P.,
et al. (2018). Age-Related Changes in the Gut Microbiota Influence Systemic
Inflammation and Stroke Outcome. Ann. Neurol. 84, 23–36. doi: 10.1002/
ana.25250 Copyright © 2021 Gu, Chen, Sun, Li, Chen, Zhou and Zhang. This is an open-access
article distributed under the terms of the Creative Commons Attribution License
(CC BY). The use, distribution or reproduction in other forums is permitted, provided
the original author(s) and the copyright owner(s) are credited and that the original
publication in this journal is cited, in accordance with accepted academic practice. No
use, distribution or reproduction is permitted which does not comply with these terms. Tan C., Wang H., Gao X., Xu R., Zeng X., Cui Z., et al. (2020). Dynamic Changes
and Prognostic Value of Gut Microbiota-Dependent Trimethylamine-N-Oxide
in Acute Ischemic Stroke. Front. Neurol. 11, 29. doi: 10.3389/fneur.2020.00029 October 2021 | Volume 11 | Article 669322 Frontiers in Cellular and Infection Microbiology | www.frontiersin.org 12
|
https://openalex.org/W3093391580
|
https://hal.science/hal-03185824/file/cp-17-529-2021.pdf
|
English
| null |
Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2
|
Climate of the past
| 2,021
|
cc-by
| 10,112
|
Mid-Pliocene Atlantic Meridional Overturning Circulation
simulated in PlioMIP2 Lunt16, Wing-Le Chan17, and
Ayako Abe-Ouchi17 1Department of Atmospheric Science, School of Environmental studies, China University of Geoscience,
Wuhan 430074, China 1Department of Atmospheric Science, School of Environmental studies, China University of Geoscience,
Wuhan 430074, China 2NORCE Norwegian Research Centre, Bjerknes Centre for Climate Research, 5007 Bergen, Norway
3Centre for Early Sapiens Behaviour, 5007 Bergen, Norway 2NORCE Norwegian Research Centre, Bjerknes Centre for Climate Research, 5007 Bergen, Norway
3Centre for Early Sapiens Behaviour, 5007 Bergen, Norway 4Department of Earth Science and Bjerknes Centre for Climate Research, University of Bergen, 5007 Bergen, Norway
5Key Laboratory of Cenozoic Geology and Environment, Institute of Geology and Geophysics, 4Department of Earth Science and Bjerknes Centre for Climate Research, University of Bergen, 5007 Bergen, Norway
5Key Laboratory of Cenozoic Geology and Environment, Institute of Geology and Geophysics, 6Laboratoire des Sciences du Climat et de l’Environnement, LSCE/IPSL, CEA-CNRS-UVSQ, Université Paris-Saclay,
91191 Gif-sur-Yvette, France 6Laboratoire des Sciences du Climat et de l’Environnement, LSCE/IPSL, CEA-CNRS-UVSQ, Université Paris-Saclay,
91191 Gif-sur-Yvette, France
7 91191 Gif-sur-Yvette, France 7Department of Geosciences, University of Connecticut, Storrs, USA
8 7Department of Geosciences, University of Connecticut, Storrs, USA p
,
y
,
,
8Climate and Global Dynamics Laboratory, National Center for Atmospheric Research, Boulder, USA
9D
f Ph
i
U i
i
f T
T
C
d d Global Dynamics Laboratory, National Center for Atmospheric Research, Boulder, USA 8Climate and Global Dynamics Laboratory, National Center for Atmospheric Research, Boulder, U
9 8Climate and Global Dynamics Laboratory, National Center for Atmo
9 9Department of Physics, University of Toronto, Toronto, Canada
10 10Institute for Marine and Atmospheric research Utrecht (IMAU), Department of Physics, Utrecht University, Utrecht, the Netherlands Utrecht University, Utrecht, the Netherlands 11Alfred Wegener Institute – Helmholtz Centre for Polar and Marine Research, Bremerhaven, Germany
12 g
,
,
y
12Institute for Environmental Physics, University of Bremen, Bremen, Germany
13Department of Physical Geography and Bolin Centre for Climate Research, Stockholm University, Sto Institute for Environmental Physics, University of Bremen, Bremen, Germany 12Institute for Environmental Physics, University of Bremen, Bremen, Germany
13Department of Physical Geography and Bolin Centre for Climate Research, Sto 12Institute for Environmental Physics, University of Bremen, Bremen, Germany
13Department of Physical Geography and Bolin Centre for Climate Research, Stockholm University, Stockholm, Sweden Institute for Environmental Physics, University of Bremen, Bremen, Germany
13Department of Physical Geography and Bolin Centre for Climate Research, Stockholm University, Stockholm, Swede 13Department of Physical Geography and Bolin Centre for Climate Research, Stockholm University, Stockholm, Sweden
14CCSR/GISS Col mbia Uni ersit
Ne
York USA 13Department of Physical Geography and Bolin Centre for Climate Research, Stockholm University, Stockholm, Sweden
14CCSR/GISS, Columbia University, New York, USA 14CCSR/GISS, Columbia University, New York, USA 15School of Earth and Environment, University of Leeds, Leeds, UK
6 16School of Geographical Sciences, University of Bristol, Bristol, UK
1 17Atmosphere and Ocean Research Institute (AORI), University of Tokyo, Kashiwa, Japan Correspondence: Zhongshi Zhang (zhongshi.zhang@cug.edu.cn) Correspondence: Zhongshi Zhang (zhongshi.zhang@cug.edu.cn) Received: 10 September 2020 – Discussion started: 22 September 2020
Revised: 4 December 2020 – Accepted: 14 January 2021 – Published: 25 February 2021 ever, there is no consistent response in the simulated Atlantic
ocean heat transport nor in the depth of the Atlantic over-
turning cell. Clim. Past, 17, 529–543, 2021
https://doi.org/10.5194/cp-17-529-2021
© Author(s) 2021. This work is distributed under
the Creative Commons Attribution 4.0 License. Clim. Past, 17, 529–543, 2021
https://doi.org/10.5194/cp-17-529-2021
© Author(s) 2021. This work is distributed under
the Creative Commons Attribution 4.0 License. Mid-Pliocene Atlantic Meridional Overturning Circulation
simulated in PlioMIP2 Zhongshi Zhang1, Xiangyu Li1, Chuncheng Guo2, Odd Helge Otterå2,3, Kerim H. Nisancioglu4, Ning Tan5,
Camille Contoux6, Gilles Ramstein6, Ran Feng7, Bette L. Otto-Bliesner8, Esther Brady8, Deepak Chandan9,
W. Richard Peltier9, Michiel L. J. Baatsen10, Anna S. von der Heydt10, Julia E. Weiffenbach10, Christian Stepanek11,
Gerrit Lohmann11,12, Qiong Zhang13, Qiang Li13, Mark A. Chandler14, Linda E. Sohl14, Alan M. Haywood15,
Stephen J. Hunter15, Julia C. Tindall15, Charles Williams16, Daniel J. 1
Introduction The mid-Piacenzian warm period (mPWP; 3.264–3.025 Ma)
was a recent period of sustained warmth in geological his-
tory, with the land–sea distribution, topography and levels
of greenhouse gases being comparable to today (Dowsett
et al., 2010, 2016; Haywood et al., 2010, 2016a). The esti-
mated global mean temperature during the mPWP was 2–
4 ◦C higher than the pre-industrial level (e.g. Dowsett et al.,
2010, 2016; Haywood et al., 2010, 2016a), and the atmo-
spheric CO2 level was above 400 ppmv (Badger et al., 2013). Thus, the mPWP climate is often thought of as a plausible
test case that has the potential to provide insights for our fu-
ture climate (e.g. Zubakov and Borzenkova, 1988; Haywood
et al., 2016b; Burke et al., 2018). In this study, we investigate the simulated AMOC in
PlioMIP2 in order to further address the question of whether
an intensified AMOC and enhanced Atlantic OHT can ex-
plain the reconstructed North Atlantic–Arctic sea surface
warming during the mPWP. In Sect. 2, we briefly introduce
the models that participated in PlioMIP2. In Sect. 3, we
compare the simulated AMOC and Atlantic OHT between
PlioMIP1 and PlioMIP2. In Sect. 4, we investigate the rela-
tionship between the simulated AMOC response and changes
in North Atlantic sea surface temperature (SST). Finally, the
results are discussed and summarized in Sect. 5. ,
;
,
)
To understand the mPWP climate, the Pliocene Modelling
Intercomparison Project (PlioMIP) Phase 1 was launched
in 2010 (Haywood et al., 2010). The major forcing consid-
ered in PlioMIP1 was an increase (compared with the pre-
industrial level) in the atmospheric CO2 level to 405 ppmv,
combined with a modern land–sea distribution (Haywood
et al., 2013). The PlioMIP1 simulations (e.g. Chan et al.,
2011; Bragg et al., 2012; Contoux et al., 2012; Kamae and
Ueda, 2012; Stepanek and Lohmann, 2012; Zhang et al.,
2012; Chandler et al., 2013; Rosenbloom et al., 2013)
showed that the global annual mean surface air tempera-
ture (SAT) was 1.9–3.6 ◦C warmer than the pre-industrial
level in the multi-model ensemble mean (Haywood et al.,
2013), whereas the strength of Atlantic Meridional Over-
turning Circulation (AMOC) was similar to the pre-industrial
level (Zhang et al., 2013a). However, when compared to ma-
rine (Dowsett et al., 2012, 2013) and terrestrial reconstruc-
tions (Salzmann et al. Mid-Pliocene Atlantic Meridional Overturning Circulation
simulated in PlioMIP2 The models show a large spread in the simulated
AMOC maximum, the Atlantic ocean heat transport and the
surface warming in the North Atlantic. Although a few mod-
els simulate a surface warming of ∼8–12 ◦C in the North At-
lantic, similar to the reconstruction from Pliocene Research,
Interpretation and Synoptic Mapping (PRISM) version 4, Abstract. In the Pliocene Model Intercomparison Project
Phase 2 (PlioMIP2), coupled climate models have been used
to simulate an interglacial climate during the mid-Piacenzian
warm period (mPWP; 3.264 to 3.025 Ma). Here, we compare
the Atlantic Meridional Overturning Circulation (AMOC),
poleward ocean heat transport and sea surface warming in
the Atlantic simulated with these models. In PlioMIP2, all
models simulate an intensified mid-Pliocene AMOC. How- Published by Copernicus Publications on behalf of the European Geosciences Union. Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 530 most models appear to underestimate this warming. The large
model spread and model–data discrepancies in the PlioMIP2
ensemble do not support the hypothesis that an intensifica-
tion of the AMOC, together with an increase in northward
ocean heat transport, is the dominant mechanism for the mid-
Pliocene warm climate over the North Atlantic. state-of-the-art boundary conditions from the Pliocene Re-
search, Interpretation and Synoptic Mapping (PRISM) ver-
sion 4 (Dowsett et al., 2016a) and focuses on the KM5c
interglacial period (3.205 Ma) during the mPWP (Haywood
et al., 2016a). The PRISM4 boundary conditions include re-
constructed ocean bathymetry and land–ice surface topog-
raphy, and they also incorporate Pliocene soils and lakes
(Dowsett et al., 2016; Haywood et al., 2016a). The most im-
portant change in boundary conditions in the northern high
latitudes is the closure of the Arctic gateways, including the
Canadian Archipelago and the Bering Strait (Haywood et al.,
2016a). In PlioMIP2, the simulated global annual mean SAT
increases by 1.7–5.2 ◦C relative to the pre-industrial level,
with a multi-model mean SAT increase of 3.2 ◦C (Haywood
et al., 2020). In the Arctic, the simulated annual mean SAT
increases by 3.7–11.6 ◦C compared with the pre-industrial
level, with a multi-model mean increase of 7.2 ◦C (de Nooi-
jer et al., 2020). 1
Introduction 2013), there was a large model–data
discrepancy (Haywood et al., 2013) in the North Atlantic and
the land realm of the Northern Hemisphere. The PlioMIP1-
simulated surface warming in the North Atlantic is ∼4–6 ◦C
smaller than the reconstruction. Because the PlioMIP1 sim-
ulations (Zhang et al., 2013a, b) did not support a stronger
Pliocene AMOC (compared with the pre-industrial level) and
an inferred enhancement of Atlantic northward ocean heat
transport (OHT) suggested by proxies (Dowsett et al., 1992;
Raymo et al., 1996), it was difficult to explain the recon-
structed strong surface warming in the high-latitude North
Atlantic during the mid-Pliocene. 2
Introduction of models used in PlioMIP2 In this study, we analyse simulations with the 15 models
that have participated and provided the simulated AMOC re-
sults to PlioMIP2 (Table 1). All 15 models have performed
simulations according to the PlioMIP2 experimental proto-
col (Haywood et al., 2016asuggested by proxies). They pro-
vide the pre-industrial control experiment (pi-E280) and the
mid-Pliocene experiment (midPliocene-Eoi400) as a mini-
mum. In the mid-Pliocene experiment, a land–sea mask with
the Arctic gateways closed and an atmospheric CO2 level
of 400 ppmv are used. The atmospheric CO2 level is in
line with the very latest high-resolution proxy reconstruc-
tion based on Boron isotopes for ∼3.2 Ma (Chalk et al.,
2018). More details on the individual models and experimen-
tal design are introduced in a recent synthesis study (Hay-
wood et al., 2020) and several individual modelling stud-
ies (Chandan and Peltier, 2017, 2018; Hunter et al., 2019;
Chan and Abe-Ouchi, 2020; Feng et al., 2020; Li et al.,
2020; Lurton et al., 2020; Stepanek et al., 2020; Tan et al.,
2020; Zhang et al., 2020). In addition to these 15 models,
MRI-CGCM (Kamae et al., 2016) and HadGEM3-GC31-
LL have taken part in PlioMIP2. However, MRI-CGCM and
HadGEM3-GC31-LL are not considered in detail here, be- To further understand the mPWP climate and to improve
upon the model–data discrepancy, PlioMIP Phase 2 was
initiated (Haywood et al., 2016a). PlioMIP2 employs the Clim. Past, 17, 529–543, 2021 https://doi.org/10.5194/cp-17-529-2021 Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 531 Table 1. Comparison of PlioMIP2 models. “PI” denotes pre-industrial, “MP” denotes mid-Pliocene and “OHT” denotes ocean heat transport. Model ID
Ocean resolution
Background vertical/
Integrated length/mean
Max AMOC
OHTa
OHTb
Reference
lat × long
diapycnal mixing
(years)
PI
MP
PI
MP
(%)
(%)
CCSM4
0.27–0.54◦× 1.1◦,
Default KPP schemec,
> 1000/100
1100/100
26.6
29.6
11
−7
−6
Feng et
L60 depth
k = 0.16 cm2 s−1 and
al. 2
Introduction of models used in PlioMIP2 (2020)
latitudinally varying
CCSM4-UoT
0.27–0.54◦× 1.1◦,
Modified KPP schemed,
4630/30
1250/30
22.6
23.5
4
9
9
Chandan and
L60 depth
identical k for PI and MP,
Peltier
k from 0.16 to 1 cm2 s−1
(2017, 2018)
and depth dependent
CCSM4-Utrecht
0.27–0.54◦×1.1◦,
Modified KPP schemed,
3100/100
2048/100
19.8
21.9
11
6
5
L60 depth
uniform k = 0.16 cm2 s−1
for PI, k from 0.1
to 1 cm2 s−1 depth
dependent for MP
CESM1.2
0.27–0.54◦× 1.1◦,
Default KPP scheme
> 1000/100
1200/100
26.7
27.0
1
−10
−9
Feng et
L60 depth
al. (2020)
CESM2
0.27–0.54◦× 1.1◦,
Default KPP scheme
1200/100
1500/100
23.0
27.8
21
−4
−5
Feng et
L60 depth
with Langmuir
al. (2020)
parameterization
COSMOS
∼3.0◦× 1.8◦,
k = 0.105 cm2 s−1
1950/100
1950/100
16.0
19.4
21
15
19
Stepanek et
L40 depth
al. (2020)
EC-Earth3-LR
1.0◦× 1.0◦,
k = 0.12 cm2 s−1
1500/100
1600/100
16.8
20.0
19
39
28
Zhang et
L75 depth
al. (2020)
GISS-E2-1-G
1◦× 1.25◦,
KPP with non-local fluxes,
5000/100
3100/100
28.2
35.1
24
4
−1
L32 depth
k = 0.10 cm2 s−1
HadCM3
1.25◦× 1.25◦,
k = 0.10 cm2 s−1
2999/100
2499/100
15.4
20.7
34
38
30
Hunter et
L20 depth
al. (2019)
IPSL-CM5A2-LR
0.5–2◦×2◦,
Function of turbulent
1500/100
3480/100
11.1
17.0
53
29
39
Tan et
L31 depth
kinetic energy
al. (2020)
IPSL-CM5A-LR
0.5–2◦×2◦,
Function of turbulent
> 800/100
3680/100
10.2
14.8
45
43
36
Tan et
L31 depth
kinetic energy
al. (2020)
IPSL-CM6A-LR
1.0◦× 1.0◦,
Turbulent kinetic energy
1100/100
1450/100
12.7
15.8
24
16
29
Lurton et
refined at 1/3◦
scheme and an
al. (2020)
in the tropics,
energy-constrained
L75 depth
parameterization of
mixing due to
internal tides
MIROC4m
0.56–1.4◦×1.4◦,
k from 0.10 to 3 cm2 s−1,
2220/100
3000/100
19.6
20.2
3
−10
−10
Chan and
L43 sigma/depth
latitudinally varying
Abe-Ouchi
(2020)
NorESM1-F
∼1.0◦× 1.0◦,
k = 0.10 cm2 s−1,
2000/100
500/100
24.5
28.1
15
1
4
Li et
L53 sigma
latitudinally varying
al. (2020)
NorESM-L
∼3.0◦× 3.0◦,
k = 0.10 cm2 s−1,
2200/100
1200/100
21.3
23.3
9
−13
−17
Li et
L32 sigma
latitudinally varying
al. (2020)
a North Atlantic ocean heat transport between 30 and 80◦N. b Atlantic ocean heat transport between 30◦S and 80◦N. c KPP (K-Profile Parameterization) scheme parameterizes boundary layer mixing and
internal diabatic mixing by convection, shear instability, internal waves, tides and double diffusion. a North Atlantic ocean heat transport between 30 and 80◦N. b Atlantic ocean heat transport between 30◦S and 80◦N. c KPP (K-Profile Parameterization) scheme parameterizes boundary layer mixing and
internal diabatic mixing by convection, shear instability, internal waves, tides and double diffusion. d KPP parameterization but with the overflow parameterization and the tidal mixing switched off.
CESM2, EC-Earth3-LR, GISS-E2-1-G and IPSL-CM6A-LR take part in the Coupled Model Intercomparison Project (CMIP) Phase 6. 4
Simulated North Atlantic sea surface warming In PlioMIP2, the simulated mid-Pliocene global annual mean
SST is between 1.2 and 4.0 ◦C warmer than the pre-industrial
level. Most models show that the strongest sea surface
warming appears in the mid- to high-latitude North Atlantic
(Figs. 4, 5). The median of the multi-model ensemble shows
that the SST increases by ∼2–8 ◦C in the North Atlantic be-
tween 30 and 80◦N (Fig. 6). The largest increase in the en-
semble median of 6–8 ◦C appears in the Labrador Sea south
of Cape Farewell (the southernmost point of Greenland). EC-Earth3-LR simulates the largest increase in the North
Atlantic SST above 12 ◦C in the mid-Pliocene experiment
(Fig. 4). In PlioMIP2, the models show that the maximum AMOC
is enhanced by 1 % to 53 % in the mid-Pliocene, relative to
the pre-industrial level (Table 1, Fig. 1). The median value of
the enhancement in maximum AMOC is 19 %. Seven mod-
els (CCSM-UoT, COSMOS, GISS-E2-1-G, HadCM3, IPSL-
CM5A-LR, IPSL-CM5A2-LR and IPSL-CM6A-LR) show
small changes in the mean depth of the AMOC cell (the
mean depth of positive streamfunction) in the mid-Pliocene
(with depth changes of less than 100 m), when compared
with the pre-industrial level. However, five models (CCSM4,
CESM1.2, CESM2, EC-Earth3-LR and MIROC4m) simu-
late a shoaling of the Atlantic overturning cell for the mid-
Pliocene, with a shoaling of ∼1190, ∼1330, ∼820, ∼350
and ∼440 m respectively. On the other hand, three models
(CCSM4-Utrecht, NorESM1-F and NorESM-L) simulate a
deeper mid-Pliocene Atlantic overturning cell with respec-
tive increases in the depth of ∼540, ∼1590 and ∼1330 m
(Figs. 1, 2). However, the SST increases in the North Atlantic (aver-
aged between 30 and 80◦N), in response to the changes
in the AMOC maximum and North Atlantic OHT (aver-
aged between 30 and 80◦N), are highly model dependent
(Fig. 5). Of the 15 PlioMIP2 models, 11 models simulate a
mean SST increase between 2 and 4 ◦C in the North Atlantic. The ranges of the changes in the AMOC maximum (from
1 % to 53 %) and mean North Atlantic OHT (from −13 %
to 43 %) are large. Meanwhile, EC-Earth3-LR produces an
increase of ∼8 ◦C in the mean North Atlantic SST, which
is associated with an intensification of 3.2 Sv (19 %) in the Compared with PlioMIP1 (Zhang et al., 2013a), the simu-
lated AMOC responses to Pliocene boundary conditions are
different in PlioMIP2 (Fig. 2). 2
Introduction of models used in PlioMIP2 d KPP parameterization but with the overflow parameterization and the tidal mixing switched off. Table 1. Comparison of PlioMIP2 models. “PI” denotes pre-industrial, “MP” denotes mid-Pliocene and “OHT” denotes ocean heat transport. els. “PI” denotes pre-industrial, “MP” denotes mid-Pliocene and “OHT” denotes ocean heat transport. Clim. Past, 17, 529–543, 2021 https://doi.org/10.5194/cp-17-529-2021 Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP 532 turning cell. However, in PlioMIP2, there is a consistent in-
crease in the maximum strength of the AMOC, whereas there
is no consistent change in the depth of Atlantic overturning
cell. cause MRI-CGCM did not provide the AMOC results to
the PlioMIP2 database, and HadGEM3-GC31-LL did not
use the enhanced land–sea distribution condition with the
Arctic gateways closed instead using the modern land–sea
distribution. Note that five models come from the Com-
munity Climate System Model/Community Earth System
Model (CCSM/CESM) family in the PlioMIP2 ensemble. To
avoid these models taking undue weights in the PlioMIP2 en-
semble, median instead of mean values are used in this study. cause MRI-CGCM did not provide the AMOC results to
the PlioMIP2 database, and HadGEM3-GC31-LL did not
use the enhanced land–sea distribution condition with the
Arctic gateways closed instead using the modern land–sea
distribution. Note that five models come from the Com-
munity Climate System Model/Community Earth System
Model (CCSM/CESM) family in the PlioMIP2 ensemble. To
avoid these models taking undue weights in the PlioMIP2 en-
semble, median instead of mean values are used in this study. 3.1
Simulated AMOC in PlioMIP2 The PlioMIP2 models produce reasonable simulations for
the pre-industrial AMOC. The pre-industrial modelled
AMOC maximums (the maximum of the Atlantic merid-
ional overturning streamfunction) range from ∼10 to 28 Sv
(1 Sv = 106 m3 s−1; Table 1, Fig. 1). The multi-model median
value of the AMOC maximums is 19.8 Sv, which is compa-
rable to the observational AMOC strength of 18.7 ± 2.1 Sv
at 26.5◦N (Kanzow et al. 2010). The depths of the Atlantic
overturning cell range from 2300 to 3800 m. 3.2
Simulated Atlantic OHT in PlioMIP2 As expected from the intensified AMOC, most models simu-
late an enhanced Atlantic OHT (averaged between 30◦S and
80◦N) in the mid-Pliocene experiments relative to the pre-
industrial level (Table 1, Fig. 3). The increases range from
4 % to 39 %. The largest enhancement is found in the simula-
tion with IPSL-CM5A2-LR, whereas the smallest enhance-
ment is simulated with NorESM1-F. In contrast, six mod-
els (CCSM4, CESM1.2, CESM2, GISS-E2-1-G, MIROC4m
and NorESM-L) show a decrease (ranging from −1 % to
−17 %) in Atlantic OHT. Of the 15 PlioMIP2 models used here, 6 of them also took
part in PlioMIP1. They are CCSM4, COSMOS, HadCM3,
IPSL-CM5A-LR, MIROC4m and NorESM-L. However, all
of these six models have submitted new pre-industrial control
experiments to the PlioMIP2 database. CCSM4 has also been
employed in a modified form by other modelling groups and
is referred to herein as CCSM4-UoT and CCSM4-Utrecht. Therefore, the pre-industrial AMOC maximums and depths
in PlioMIP2 are slightly different to the values in PlioMIP1. Obviously, there is no linear relationship between the in-
tensification in the AMOC and the changes in mean At-
lantic OHT in the PlioMIP2 simulations (Fig. 2b). For ex-
ample, GISS-E2-1-G and IPSL-CM6A-LR both simulate in-
creases of 24 % in the AMOC maximum; however, GISS-E2-
1-G shows a −1 % decrease in mean Atlantic OHT, whereas
IPSL-CM6A-LR shows an increase of 29 %. CCSM4 and
CCSM4-Utrecht also show the same increase of 11 % in the
AMOC maximum but inverse responses in the mean Atlantic
OHT. This large model spread in PlioMIP2 suggests that the
relationship between the AMOC strength and Atlantic north-
ward OHT are highly model dependent. 4
Simulated North Atlantic sea surface warming In PlioMIP1, there was no
consistent increase in the maximum strength of the AMOC,
whereas there was a consistent shoaling of the Atlantic over- Clim. Past, 17, 529–543, 2021 https://doi.org/10.5194/cp-17-529-2021 Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 ne Atlantic Meridional Overturning Circulation simulated in PlioMIP2 533 e simulated AMOC (unit: Sv) in PlioMIP2. “PI” denotes pre-industrial, and “MP” denotes mid-Pliocene. Figure 1. The simulated AMOC (unit: Sv) in PlioMIP2. “PI” denotes pre-industrial, and “MP” denotes mid-Pliocene. OHT changes by 0.06 PW (9 %), 0.04 PW (6 %), −0.02 PW
(−4 %) respectively. AMOC maximum and an enhancement of 0.16 PW (41 %)
in the mean North Atlantic OHT. CCSM4-UoT, CCSM4-
Utrecht and CESM2 produce a similar increase of ∼5 ◦C in
the mean North Atlantic SST, while the intensification in the
AMOC maximum shows a large range covering 0.9 Sv (4 %),
2.1 Sv (11 %) and 4.7 Sv (21 %) and the mean North Atlantic AMOC maximum and an enhancement of 0.16 PW (41 %)
in the mean North Atlantic OHT. CCSM4-UoT, CCSM4-
Utrecht and CESM2 produce a similar increase of ∼5 ◦C in
the mean North Atlantic SST, while the intensification in the
AMOC maximum shows a large range covering 0.9 Sv (4 %),
2.1 Sv (11 %) and 4.7 Sv (21 %) and the mean North Atlantic In
PlioMIP2,
the
surface
warming
simulated
with
CCSM4-UoT, CCSM4-Utrecht, CESM2 and EC-Earth3-LR
is close to or warmer than the PRISM4 reconstructions (Fo-
ley and Dowsett, 2019) in the North Atlantic between 30 and https://doi.org/10.5194/cp-17-529-2021 Clim. Past, 17, 529–543, 2021 534
Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2
Figure 2. Simulated changes in the AMOC maximum, depth and Atlantic northward OHT. (a) Changes in the AMOC maximum (unit: %)
vs. responses in the mean depth of the AMOC cell (unit: m). (b) Changes in the AMOC maximum (unit: %) vs. responses in the mean ocean
heat transport in the Atlantic between 30◦S and 80◦N (unit: %). The blue markers show the PlioMIP1 simulations, and the red markers show
the PlioMIP2 simulations. The vertical and horizontal lines show the model range, and the intersection of these lines indicates the median
value. Note that only the mean values of the AMOC maximum, depth and Atlantic northward OHT for each model are used here to calculate
the anomalies, and significance tests are not employed. . 4
Simulated North Atlantic sea surface warming Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 34
Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 534 Figure 2. Simulated changes in the AMOC maximum, depth and Atlantic northward OHT. (a) Changes in the AMOC maximum (unit: %)
vs. responses in the mean depth of the AMOC cell (unit: m). (b) Changes in the AMOC maximum (unit: %) vs. responses in the mean ocean
heat transport in the Atlantic between 30◦S and 80◦N (unit: %). The blue markers show the PlioMIP1 simulations, and the red markers show
the PlioMIP2 simulations. The vertical and horizontal lines show the model range, and the intersection of these lines indicates the median
value. Note that only the mean values of the AMOC maximum, depth and Atlantic northward OHT for each model are used here to calculate
the anomalies, and significance tests are not employed. Figure 3. Simulated Atlantic poleward oceanic heat transport in PlioMIP2 (unit: PW). Blue dashed lines show the pre-industrial, and red
solid lines show the mid-Pliocene. Figure 3. Simulated Atlantic poleward oceanic heat transport in PlioMIP2 (unit: PW). Blue dashed lines show the pre-industrial, and red
solid lines show the mid-Pliocene. Figure 3. Simulated Atlantic poleward oceanic heat transport in PlioMIP2 (unit: PW). Blue dashed lines show the pre-industrial, and red
solid lines show the mid-Pliocene. https://doi.org/10.5194/cp-17-529-2021 Clim. Past, 17, 529–543, 2021 Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 535 mulated mid-Pliocene annual SST anomalies in PlioMIP2 (unit: ◦C). µ denotes the global mean. Figure 4. Simulated mid-Pliocene annual SST anomalies in PlioMIP2 (unit: ◦C). µ denotes the global mean Clim. Past, 17, 529–543, 2021 https://doi.org/10.5194/cp-17-529-2021 536
Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2
Figure 5. Simulated changes in the AMOC maximum and the North Atlantic (nAtlantic) OHT, and responses in the high-latitude North
Atlantic SST. The North Atlantic OHT is the averaged value between 30 and 80◦N. The high-latitude North Atlantic includes the Atlantic
and Greenland–Iceland–Norwegian (GIN) seas between 30 and 80◦N. Note that only the mean values of the AMOC maximum, the North
Atlantic OHT and the SST for each model are used to calculate the anomalies, and significance tests are not employed. 36
Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP . 4
Simulated North Atlantic sea surface warming Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 536 Figure 5. Simulated changes in the AMOC maximum and the North Atlantic (nAtlantic) OHT, and responses in the high-latitude North
Atlantic SST. The North Atlantic OHT is the averaged value between 30 and 80◦N. The high-latitude North Atlantic includes the Atlantic
and Greenland–Iceland–Norwegian (GIN) seas between 30 and 80◦N. Note that only the mean values of the AMOC maximum, the North
Atlantic OHT and the SST for each model are used to calculate the anomalies, and significance tests are not employed. 80◦N, whereas the other models still appear to underestimate
the North Atlantic SST (Fig. 6). Previous studies (Brierley
and Fedorov, 2016; Otto-Bliesner et al., 2017; Song et al.,
2018) showed that the closing of the Arctic gateways led to
warmer North Atlantic SSTs in the mid-Pliocene experiment,
when compared to the pre-industrial level. However, the Arc-
tic gateways are closed in all PlioMIP2 simulations analysed
here, but not all of them simulate the warm North Atlantic
SSTs as reconstructed in the PRISM4 data set (Foley and
Dowsett, 2019). Although the Arctic gateways may lead to a
better agreement between simulated and reconstructed mid-
Pliocene North Atlantic SSTs in some models, the effect is either not present for all of the models or it is not of suffi-
cient amplitude to fully resolve the model–data discord. The
PlioMIP2 models show a larger model spread in the simu-
lated mid-Pliocene SST increases in the high-latitude North
Atlantic, as well as the responses in the AMOC and North
Atlantic OHT, relative to PlioMIP1. This reduced agreement
is not surprising, as the model spread in global average sur-
face temperatures is likewise more pronounced in PlioMIP2:
1.86–3.60 ◦C in PlioMIP1 (Haywood et al., 2013) compared
with 1.7–5.2 ◦C in PlioMIP2 (Haywood et al., 2020). https://doi.org/10.5194/cp-17-529-2021 Clim. Past, 17, 529–543, 2021 ang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2
53
Figure 6. PlioMIP2 and PRISM4 SST comparison in the Atlantic. (a) PRISM4 SST anomalies (points) at data sites in the Atlantic a
the Mediterranean plotted against the multi-model ensemble median of SST anomalies (shaded; the mid-Pliocene vs. the pre-industri
simulated in PlioMIP2 (unit: ◦C). 5
Discussion and summary With COSMOS, a sen-
sitivity experiment forced with the modern land–sea dis-
tribution (the Arctic gateways open) also shows a weaker
AMOC, when compared with the core mid-Pliocene simu-
lation (Stepanek et al., 2020). As revealed in the earlier study
(Otto-Bliesner et al., 2017), closed Arctic gateways lead to a
stronger AMOC by inhibiting Arctic freshwater export to the
North Atlantic. However, the magnitude of the intensification
in the AMOC due to the closed Arctic gateways seems highly
model dependent. Some simulations suggest that the AMOC
is enhanced by ∼2 Sv due to the closed Bering Strait (Brier-
ley and Fedorov, 2016; Otto-Bliesner et al., 2017; Song et al.,
2018), whereas some unpublished simulations in PlioMIP2
show much larger responses. Without consistent sensitivity
experiments for the Arctic gateways, it remains difficult to
reveal the range of model spread on the gateways’ impacts in
PlioMIP2. This model dependence will be addressed in more
dedicated sensitivity experiments in the future. g
y
It should be noted that observations of strong high-latitude
warming in the North Atlantic are not sufficient to constrain
the strength of AMOC or OHT (Zhang et al., 2013b). The
AMOC strength measures the contrast in water transport be-
tween the upper and lower branches of the Atlantic cells, but
the OHT is also influenced by the contrast in water temper-
ature as well as the depth of the AMOC. Moreover, OHT
can be decomposed into a (vertical) MOC component and
a (horizontal) gyre component. While the MOC component
dominates in most of the Atlantic region, the gyre component
has a comparable magnitude in the subpolar region (Williams
et al., 2015). Therefore, there is no one-to-one correspon-
dence between AMOC and OHT, especially in the subpolar
regions. Furthermore, the SST warming pattern is not en-
tirely determined by OHT, as demonstrated by the simula-
tions in both PlioMIP1 and PlioMIP2. y
p
In PlioMIP2, the large model spread does not support the
notion that an intensified mid-Pliocene AMOC is the princi-
pal mechanism responsible for the simulated warming of the
North Atlantic SSTs. Compared with CCSM4, both CCSM4-
UoT and CCSM4-Utrecht (Table 1) simulate warmer SSTs
in the North Atlantic, suggesting that the increased back-
ground ocean vertical mixing parameters likely contribute to
the strong mid-Pliocene North Atlantic warming simulated
with these two models. Each model’s climate sensitivity also
influences the simulated mid-Pliocene warming in PlioMIP2. 5
Discussion and summary the mid-Pliocene global mean SST (SAT) of 3.8 ◦C (5.1 ◦C)
relative to the pre-industrial level, which is the second largest
warming in PlioMIP2 (Fig. 4). Moreover, a new lake and soil
condition is employed in PlioMIP2 (Pound et al., 2014; Hay-
wood et al., 2016a). Methods for modifying the soil condi-
tion and their impacts on climate in the models are highly
model dependent due to the large variety of land surface
schemes included in the PlioMIP2 models, which could fur-
ther amplify the diversity of warming signals in high-latitude
regions. As not all models carried out the sensitivity exper-
iments designed in PlioMIP2, it remains difficult to distin-
guish which change in boundary conditions is more dominant
for the strong mid-Pliocene North Atlantic surface warm-
ing. Earlier studies (e.g. Feng et al., 2017) noted that the
North Atlantic warming is not a unique feature in many mid-
Pliocene simulations, as the warming in the North Pacific is
also remarkable (Fig. 4). This inter-basin symmetry suggests
a potentially important component of the zonal mean polar
amplification of the SST warming across the North Atlantic. Energy balance analyses (Hill et al., 2014; Feng et al., 2017)
show that amplified zonal mean northern high-latitude warm-
ing is dominated by regional radiative feedbacks from low-
ered surface albedo and an enhanced high-latitude green-
house effect (from changes in water vapour), even with an
enhanced AMOC due to gateway closure. Compared with the PlioMIP1 ensemble in which the Arctic
gateways were kept open, all PlioMIP2 models forced with
the PRISM4 reconstructions that consider the closed Arc-
tic gateways simulate an intensification in the mid-Pliocene
AMOC. CCSM4, COSMOS, HadCM3, IPSL-CM5A-LR,
MIROC4m and NorESM-L have all participated in both
PlioMIP1 and PlioMIP 2. These six models simulate an in-
crease (compared with the pre-industrial level) in the mid-
Pliocene AMOC maximum, which is larger in PlioMIP2
than in PlioMIP1, supporting the hypothesis that closed Arc-
tic gateways are a requirement for the intensification of the
mid-Pliocene AMOC. There are several further lines of ev-
idence that support this hypothesis. HadGEM3-GC31-LL,
which carried out the mid-Pliocene experiment forced with
the PlioMIP2 boundary conditions, except with the land–sea
distribution condition identical to the pre-industrial simula-
tion, produces a weaker mid-Pliocene AMOC (with a max-
imum of 14.3 Sv) compared with the pre-industrial simula-
tion (with a maximum of 16.1 Sv). 4
Simulated North Atlantic sea surface warming (b) Black dots show the PRISM4 SST anomalies (unit: ◦C) at each site. Vertical blue lines and dots sho
the PlioMIP1 ranges and median values of changes in SST for each site. Coloured markers show SST changes simulated by each mod
in PlioMIP2. The PRISM4 SST anomalies are calculated based on the PRISM4 mid-Pliocene reconstructions (3.19–3.22 Ma; Foley a
Dowsett, 2019) and the modern observation (1870–1899; Rayner et al., 2003). 537 Figure 6. PlioMIP2 and PRISM4 SST comparison in the Atlantic. (a) PRISM4 SST anomalies (points) at data sites in the Atlantic and
the Mediterranean plotted against the multi-model ensemble median of SST anomalies (shaded; the mid-Pliocene vs. the pre-industrial)
simulated in PlioMIP2 (unit: ◦C). (b) Black dots show the PRISM4 SST anomalies (unit: ◦C) at each site. Vertical blue lines and dots show
the PlioMIP1 ranges and median values of changes in SST for each site. Coloured markers show SST changes simulated by each model
in PlioMIP2. The PRISM4 SST anomalies are calculated based on the PRISM4 mid-Pliocene reconstructions (3.19–3.22 Ma; Foley and
Dowsett, 2019) and the modern observation (1870–1899; Rayner et al., 2003). https://doi.org/10.5194/cp-17-529-2021 Clim. Past, 17, 529–543, 2021 Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 538 5
Discussion and summary This can be ex-
plained with the increased ventilation in the Southern Ocean
and does not necessarily depend on an intensified AMOC
(Zhang et al., 2013b). However, simulations of Southern
Ocean dynamics are highly model dependent (Zhang et al.,
2013a). In addition to the Southern Ocean, the Pliocene deep-
ocean circulation in the North Pacific appears different to the
present day. In the subarctic North Pacific, high accumula-
tion rates of calcium carbonate and biogenic opal suggest a
strong deep convection there and, thus, the existence of North
Pacific deep-water formation and a Pacific Meridional Over-
turning Circulation (PMOC, Burls et al., 2017). However,
with an intensified AMOC, a PMOC remains absent in the
PlioMIP2 simulations. Although the model–data discrepancy is reduced in the
North Atlantic partly due to the intensified AMOC, the
model–data mismatch remains large in other regions in
PlioMIP2, for example sites 1081, 1082, 1084 and 1087 in
the Benguela upwelling region (Fig. 6). The PRISM4 (Fo-
ley and Dowsett, 2019) and other syntheses of Pliocene SST
(Fedorov et al., 2013, McClymont et al., 2020) reconstruct
that the SSTs are about 6–8 ◦C warmer than today in this
region. All PlioMIP2 models underestimate this warming in
PlioMIP2 (Fig. 6). Even EC-Earth3-LR, which produces the
warmest mid-Pliocene simulation in the North Atlantic, only
simulates 2–4 ◦C sea surface warming in the Benguela up-
welling region. In summary, all 15 coupled models in PlioMIP2 used in
this study simulate an intensified mid-Pliocene AMOC, rel-
ative to the pre-industrial level. The simulated AMOC max-
imum (the maximum of the Atlantic meridional overturning
streamfunction) increases by between 1 % and 53 %. How-
ever, these models do not simulate a consistent change in the
depth of the Atlantic overturning cell and the Atlantic OHT. The spread in the responses of the AMOC and Atlantic OHT
in the models becomes larger in PlioMIP2, when compared
with PlioMIP1. In the North Atlantic, EC-Earth3-LR and the
models from the CCSM/CESM family can simulate an SST
increase (∼8–12 ◦C) close to the PRISM4 reconstruction,
whereas other models appear to underestimate the sea sur-
face warming. In PlioMIP2, the model–data discrepancy is
reduced in the North Atlantic, but the discrepancy remains
large in the upwelling regions. 5
Discussion and summary For example, relative to CCSM4 and CESM1.2, CESM2 has
a greater equilibrium climate sensitivity (Feng et al., 2020;
Haywood et al., 2020) and simulates the strongest North
Atlantic warming in the mid-Pliocene experiment. With the
modern land–sea distribution conditions, HadGEM3-GC31-
LL simulates a weakened mid-Pliocene AMOC but much
warmer SSTs in the North Atlantic as well as an increase in Nevertheless, the PlioMIP2 experiments simulate a sea
surface warming that is in better agreement with the PRISM4
reconstructions (Foley and Dowsett, 2019) in the North At-
lantic, relative to the PlioMIP1 ensemble. As shown in the
synthesis paper by Haywood et al. (2020), the multi-model
means (with equal weight for each model) agree well with the
reconstructions at the North Atlantic sites 609 and 1308, and
they show only small differences from the reconstructions at
sites 982 and 642. The comparison between the PlioMIP2
simulations and the SST reconstructions in the KM5c inter-
glacial (McClymont et al., 2020) also demonstrates the re-
duced model–data discord. However, the improved model–data agreement in the
North Atlantic is primarily caused by the relatively warm https://doi.org/10.5194/cp-17-529-2021 Clim. Past, 17, 529–543, 2021 Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP 539 mid-Pliocene simulations run with EC-Earth3-LR and the
five models from the CCSM/CESM family (Fig. 6). For the
other models, the range of warming at these sites is similar
to that of PlioMIP1. This large model spread suggests that
the reconstructed strong mid-Pliocene sea surface warming
in the North Atlantic is not necessarily caused by the in-
tensified AMOC and enhanced Atlantic northward OHT as
suggested previously (Dowsett et al., 1992; Raymo et al.,
1996). Even given the intensified AMOC in PlioMIP2 due
to the closed Arctic gateways, most models produce the mid-
Pliocene North Atlantic sea surface warming that is weaker
than the PRISM4 reconstruction (Foley and Dowsett, 2019). PlioMIP2, CCSM4-UoT and CCSM4-Utrecht have consid-
ered increasing the ocean background mixing parameters, but
no model has tested the impact of a reduction in the extra-
tropical cloud albedo in the mid-Pliocene experiments. This
mechanism can be further addressed in the future to inves-
tigate whether it is a suitable candidate for improving the
simulation for upwelling regions. Furthermore, it remains problematic to use the intensified
AMOC to explain other features of the mid-Pliocene ocean
circulation. During the mPWP, the vertical and meridional
δ13C gradients are reduced in the Atlantic. Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 Initiative REKLIM and the Alfred Wegener Institute’s PACES2 re-
search programme. Author contributions. ZZ and XL analysed the data and wrote
the draft of the paper. All authors contributed to discussion of the
results and writing the paper. Qiong Zhang acknowledges financial support from the Swedish
Research Council (Vetenskapsrådet; grant nos. 2013-06476 and
2017-04232). The model simulations with EC-Earth3 and the data
analysis were performed using resources provided by ECMWF’s
computing and archive facilities and the Swedish National Infras-
tructure for Computing (SNIC) at the National Supercomputer Cen-
tre (NSC), which is partially funded by the Swedish Research Coun-
cil through grant agreement no. 2016-07213. Competing interests. The authors declare that they have no con-
flict of interest. flict of interest. Special issue statement. This article is part of the special issue
“PlioMIP Phase 2: experimental design, implementation and scien-
tific results”. It is not associated with a conference. Alan M. Haywood, Stephen J. Hunter and Julia C. Tindall ac-
knowledge the FP7 Ideas programme of the European Research
Council (grant no. PLIO-ESS, 278636), the Past Earth Network
(EPSRC grant no. EP/M008.363/1) and the University of Leeds Ad-
vanced Research Computing service. Julia C. Tindall was also sup-
ported through the Centre for Environmental Modelling And Com-
putation (CEMAC), University of Leeds. Acknowledgements. Zhongshi Zhang and Xiangyu Li were sup-
ported by the National Natural Science Foundation of China (grant
no. 41888101), the National Key Research and Development Pro-
gram of China (grant no. 2018YFA0605602), the China Scholarship
Council (grant no. 201804910023), the China Postdoctoral Science
Foundation (grant no. 2015M581154), the Norwegian Research
Council (project nos. 221712, 229819 and 262618), the NordForsk-
funded project GREENICE (project no. 61841) and computing re-
sources from Notur/NorStore (project nos. NN9133/NS9133 and
NN9486/NS9486). Wing-Le Chan and Ayako Abe-Ouchi acknowledge funding
from JSPS KAKENHI (grant 17H06104) and MEXT KAKENHI
(grant 17H06323), and they are grateful to JAMSTEC for use of the
Earth Simulator supercomputer. Financial support. This research has been supported by the Na-
tional Natural Science Foundation of China (grant no. 41888101),
the National Key Research and Development Program of China
(grant no. 2018YFA0605602), the China Scholarship Council
(grant no. 201804910023), the China Postdoctoral Science Foun-
dation (grant no. 2015M581154), the Norwegian Research Coun-
cil (project nos. 221712, 229819 and 262618) and the NordForsk-
funded project GREENICE (project no. 61841). Ran Feng, Bette L. Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 Otto-Bliesner and Esther Brady acknowledge
the CESM project, which is primarily supported by the National
Science Foundation (NSF). This material is based upon work sup-
ported by the National Center for Atmospheric Research (NCAR),
which is a major facility sponsored by the NSF under coopera-
tive agreement no. 1852977. This research was additionally spon-
sored by U.S. National Science Foundation grant nos. 1903650
and 1814029 to Ran Feng and grant no. 1418411 to Bette L. Otto-Bliesner. Computing and data storage resources, including the
Cheyenne supercomputer (https://doi.org/10.5065/D6RX99HX),
were provided by the Computational and Information Systems Lab-
oratory (CISL) at NCAR. Review statement. This paper was edited by Appy Sluijs and re-
viewed by Chris Brierley and one anonymous referee. W. Richard Peltier and Deepak Chandan were supported by a
Canadian NSERC Discovery Grant (grant no. A9627), and they
wish to acknowledge the SciNet HPC consortium for providing
computing facilities. SciNet is funded by the Canada Foundation
for Innovation under the auspices of Compute Canada, the Govern-
ment of Ontario, the Ontario Research Fund – Research Excellence
and the University of Toronto. 5
Discussion and summary The large model spread and
the remaining model–data discrepancy suggests that an in-
tensified AMOC and an enhanced Atlantic northward OHT
cannot explain the reconstructed warm climate of the mid-
Pliocene surface oceans. g
g
A major feature of the mid-Pliocene seems to be the large
increase in SST (about 2–10 ◦C) in the mid-latitude coastal
upwelling regions and the relatively smaller increases in SST
(about 2–4 ◦C) in the mid- to high latitudes (Fedorov et al.,
2013) compared with the pre-industrial level, although some
studies suggest that SST reconstructions in upwelling re-
gions are highly proxy dependent (e.g. Leduc et al., 2014). For example, in the Benguela upwelling region, the Mg/Ca-
based SST is ∼3–10 ◦C colder than the alkenone-based SST
(Leduc et al., 2014). In the California upwelling region, Fo-
ley and Dowsett (2019) show that the Pliocene SST is simi-
lar to today, whereas Fedorov et al. (2013) show the regional
SST is about 2–8 ◦C warmer than today. Despite the uncer-
tainties in reconstructions, the simulated warming in the mid-
latitude upwelling regions in PlioMIP2 can be found at the
low end of the proxy-estimated range. Realistic simulations
in upwelling regions require good model abilities with re-
spect to simulating large-scale ocean stratification and sea
surface wind stress (Miller and Tziperman, 2017; Li et al.,
2019), which are partly model-resolution dependent in both
atmosphere and ocean models (Gent et al., 2010; Small et al.,
2015). Data availability. To access the PlioMIP2 database, please send
a request to Alan M. Haywood (a.m.haywood@leeds.ac.uk). PlioMIP2 data from CESM2, EC-Earth3-LR, GISS-E2-1-G, IPSL-
CM6A-LR and NorESM1-F can be obtained from the Earth System
Grid Federation (ESGF, 2020, https://esgf-node.llnl.gov/search/
cmip6/, last access: 3 December 2020). Taken together, these model–data discrepancies make it
difficult to associate the intensified AMOC and enhanced
Atlantic northward OHT with the reconstructed high mid-
Pliocene SSTs. Fedorov et al. (2013) suggested a possible
mechanism for understanding the warm SSTs during the
mPWP. Increased mixing in the subtropical ocean and re-
duced extra-tropical cloud albedo cause a strong warming
in the mid-latitudes, including some upwelling regions. In Clim. Past, 17, 529–543, 2021 https://doi.org/10.5194/cp-17-529-2021 540 Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 R.: Regional and global climate for the
mid-Pliocene using the University of Toronto version of CCSM4
and PlioMIP2 boundary conditions, Clim. Past, 13, 919–942,
https://doi.org/10.5194/cp-13-919-2017, 2017. Feng, R., Otto-Bliesner, B. L., Fletcher, T. L., Tabor, C. R., Ballan-
tyne, A. P., and Brady, E. C.: Amplified Late Pliocene terrestrial
warmth in northern high latitudes from greater radiative forcing
and closed Arctic Ocean gateways, Earth Planet. Sc. Lett., 466,
129–138, 2017. Chandan, D. and Peltier, W. R.: On the mechanisms of warm-
ing the mid-Pliocene and the inference of a hierarchy of cli-
mate sensitivities with relevance to the understanding of climate
futures, Clim. Past, 14, 825–856, https://doi.org/10.5194/cp-14-
825-2018, 2018. Feng, R., Otto-Bliesner, B. L., Brady, E. C., and Rosen-
bloom, N.: Increased Climate Response and Earth System
Sensitivity from CCSM4 to CESM2 in mid-Pliocene sim-
ulations, J. Adv. Model. Earth Sy., 12, e2019MS002033,
https://doi.org/10.1029/2019MS002033, 2020. Chandler, M. A., Sohl, L. E., Jonas, J. A., Dowsett, H. J., and Kel-
ley, M.: Simulations of the mid-Pliocene Warm Period using
two versions of the NASA/GISS ModelE2-R Coupled Model,
Geosci. Model Dev., 6, 517–531, https://doi.org/10.5194/gmd-6-
517-2013, 2013. Foley, K. M. and Dowsett, H. J.: Community sourced mid-
Piacenzian sea surface temperature (SST) data, US Geological
Survey data release, https://doi.org/10.5066/P9YP3DTV, 2019. Contoux, C., Ramstein, G., and Jost, A.: Modelling the mid-
Pliocene Warm Period climate with the IPSL coupled model and
its atmospheric component LMDZ5A, Geosci. Model Dev., 5,
903–917, https://doi.org/10.5194/gmd-5-903-2012, 2012. Gent, P. R., Yeager, S. G., Neale, R. B., Levis, S., and Bailey, D. A.:
Improvements in a half degree atmosphere/land version of the
CCSM, Clim. Dynam., 34, 819–833, 2010. Haywood, A. M., Dowsett, H. J., Otto-Bliesner, B., Chandler, M. A.,
Dolan, A. M., Hill, D. J., Lunt, D. J., Robinson, M. M., Rosen-
bloom, N., Salzmann, U., and Sohl, L. E.: Pliocene Model Inter-
comparison Project (PlioMIP): experimental design and bound-
ary conditions (Experiment 1), Geosci. Model Dev., 3, 227–242,
https://doi.org/10.5194/gmd-3-227-2010, 2010. de Nooijer, W., Zhang, Q., Li, Q., Zhang, Q., Li, X., Zhang, Z.,
Guo, C., Nisancioglu, K. H., Haywood, A. M., Tindall, J. C.,
Hunter, S. J., Dowsett, H. J., Stepanek, C., Lohmann, G., Otto-
Bliesner, B. L., Feng, R., Sohl, L. E., Chandler, M. A., Tan, N.,
Contoux, C., Ramstein, G., Baatsen, M. L. J., von der Heydt,
A. S., Chandan, D., Peltier, W. R., Abe-Ouchi, A., Chan, W.-L.,
Kamae, Y., and Brierley, C. Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 541 turning circulation (PMOC) during the warm Pliocene, Science
Advances, 3, e1700156, https://doi.org/10.1126/sciadv.1700156,
2017. turning circulation (PMOC) during the warm Pliocene, Science
Advances, 3, e1700156, https://doi.org/10.1126/sciadv.1700156,
2017. J., Lunt, D. J., Foley, K. M., and Riesselman, C. R.: Assessing
confidence in Pliocene sea surface temperatures to evaluate pre-
dictive models, Nat. Clim. Change, 2, 365–371, 2012. Chalk, T. B., De La Vega, E., Foster, G. L., Bysani, R., and Wil-
son, P. A.: Warming in a warm world: Orbital CO2 forcing vari-
ations in the warm Pliocene, Abstract PP21C-1434 presented at
the 2018 AGU Fall Meeting, San Francisco, CA, 11 December
2018. Dowsett, H. J., Foley, K. M., Stoll, D. K., Chandler, M. A., Sohl, L. Dowsett, H. J., Foley, K. M., Stoll, D. K., Chandler, M. A., Sohl, L. E., Bentsen, M., Otto-Bliesner, B. L., Bragg, F. J., Chan, W.-L.,
Contoux, C., Dolan, A. M., Haywood, A. M., Jonas, J. A., Jost,
A., Kamae, Y., Lohmann, G., Lunt, D. J., Nisancioglu, K. H.,
Abe-Ouchi, A., Ramstein, G., Riesselman, C. R., Robinson, M. M., Rosenbloom, N. A., Salzmann, U., Stepanek, C., Strother, S. L., Ueda, H., Yan, Q., and Zhang, Z.: Sea Surface Temperature
of the mid-Piacenzian Ocean: A Data-Model Comparison, Sci. Rep.-UK, 3, 2013, https://doi.org/10.1038/srep02013, 2013. E., Bentsen, M., Otto-Bliesner, B. L., Bragg, F. J., Chan, W.-L.,
Contoux, C., Dolan, A. M., Haywood, A. M., Jonas, J. A., Jost, A., Kamae, Y., Lohmann, G., Lunt, D. J., Nisancioglu, K. H., Abe-Ouchi, A., Ramstein, G., Riesselman, C. R., Robinson, M. Chan, W.-L. and Abe-Ouchi, A.: Pliocene Model Intercomparison
Project (PlioMIP2) simulations using the Model for Interdisci-
plinary Research on Climate (MIROC4m), Clim. Past, 16, 1523–
1545, https://doi.org/10.5194/cp-16-1523-2020, 2020. Rosenbloom, N. A., Salzmann, U., Stepanek, C., Strothe L., Ueda, H., Yan, Q., and Zhang, Z.: Sea Surface Temperature
of the mid-Piacenzian Ocean: A Data-Model Comparison, Sci. Chan, W.-L., Abe-Ouchi, A., and Ohgaito, R.: Simulating the mid-
Pliocene climate with the MIROC general circulation model:
experimental design and initial results, Geosci. Model Dev., 4,
1035–1049, https://doi.org/10.5194/gmd-4-1035-2011, 2011. ESGF: Datasets from CMIP6 simulations, available at: https://
esgf-node.llnl.gov/search/cmip6/, last access: 3 December 2020. Fedorov, A. V., Brierley, C. M., Lawrence, K. T., Liu, Z., Dekens, P. S., and Ravelo, A. C.: Patterns and mechanisms of early Pliocene
warmth, Nature, 496, 43–49, 2013. Chandan, D. and Peltier, W. References Badger, M. P. S., Schmidt, D. N., Mackensen, A., and Pan-
cost,
R. D.:
High-resolution
alkenone
palaeobarome-
try indicates relatively stable pCO2
during the pliocene
(3.3–2.8 ma),
Philos. T. R. Soc. A,
371,
20130094,
https://doi.org/10.1098/rsta.2013.0094, 2013. Anna S. von der Heydt and Michiel L. J. Baatsen, acknowl-
edge the Netherlands Earth System Science Centre (NESSC) pro-
gramme, which is financially supported by the Ministry of Educa-
tion, Culture and Science (OCW; grant no. 024.002.001). Simula-
tions with CCSM4-Utrecht were performed at the SURFsara Dutch
national computing facilities and were sponsored by NWO-EW
(Netherlands Organization for Scientific Research, Exact Sciences)
under project no. 17189. Bragg, F. J., Lunt, D. J., and Haywood, A. M.: Mid-Pliocene cli-
mate modelled using the UK Hadley Centre Model: PlioMIP
Experiments 1 and 2, Geosci. Model Dev., 5, 1109–1125,
https://doi.org/10.5194/gmd-5-1109-2012, 2012. Brierley, C. M. and Fedorov, A. V.: Comparing the impacts
of
Miocene–Pliocene
changes
in
inter-ocean
gateways
on
climate:
Central
American
Seaway,
Bering
Strait,
and
Indonesia,
Earth
Planet. Sc. Lett.,
444,
116–130,
https://doi.org/10.1016/j.epsl.2016.03.010, 2016. Gerrit Lohmann and Christian Stepanek acknowledge the use of
computational resources from the Computing and Data Centre of
the Alfred Wegener Institute – Helmholtz-Centre for Polar and Ma-
rine Research towards the generation of the COSMOS PlioMIP2
simulation ensemble. Gerrit Lohmann acknowledges funding via
the Alfred Wegener Institute’s PACES2 research programme. Chris-
tian Stepanek acknowledges funding from the Helmholtz Climate Burke, K. D., Williams, J. W., Chandler, M. A., Haywood, A. M.,
Lunt, D. J., and Otto-Bliesner, B. L.: Pliocene and Eocene pro-
vide best analogs for near-future climates, P. Natl. Acad. Sci. USA, 115, 13288–13293, 2018. Burls, N. J., Fedorov, A. V., Sigman, D. M., Jaccard, S. L., Tiede-
mann, R., and Haug G. H.: Active Pacific meridional over- Clim. Past, 17, 529–543, 2021 https://doi.org/10.5194/cp-17-529-2021 Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 A., Sohl, L. E., Otto-Bliesner, B. L., Feng, R., Brady, E. C.,
von der Heydt, A. S., Baatsen, M. L. J., and Lunt, D. J.: The
Pliocene Model Intercomparison Project Phase 2: large-scale cli-
mate features and climate sensitivity, Clim. Past, 16, 2095–2123,
https://doi.org/10.5194/cp-16-2095-2020, 2020. view from ∼3 million years ago, Clim. Past, 16, 1599–1615,
https://doi.org/10.5194/cp-16-1599-2020, 2020. view from ∼3 million years ago, Clim. Past, 16, 1599–1615,
https://doi.org/10.5194/cp-16-1599-2020, 2020. Miller, M. D. and Tziperman, E.: The effect of changes in surface
winds and ocean stratification on coastal upwelling and sea sur-
face temperatures in the Pliocene, Paleoceanography, 32, 371–
383, 2017. Otto-Bliesner, B. L., Jahn, A., Feng, R., Brady, E. C., Hu, A., and
Löfverström, M.: Amplified North Atlantic warming in the late
Pliocene by changes in Arctic gateways, Geophys. Res. Lett., 44,
957–964, 2017. Hill, D. J., Haywood, A. M., Lunt, D. J., Hunter, S. J., Bragg,
F. J., Contoux, C., Stepanek, C., Sohl, L., Rosenbloom, N. A., Chan, W.-L., Kamae, Y., Zhang, Z., Abe-Ouchi, A., Chan-
dler, M. A., Jost, A., Lohmann, G., Otto-Bliesner, B. L., Ram-
stein, G., and Ueda, H.: Evaluating the dominant components of
warming in Pliocene climate simulations, Clim. Past, 10, 79–90,
https://doi.org/10.5194/cp-10-79-2014, 2014. Pound, M. J., Tindall, J., Pickering, S. J., Haywood, A. M., Dowsett,
H. J., and Salzmann, U.: Late Pliocene lakes and soils: a global
data set for the analysis of climate feedbacks in a warmer
world, Clim. Past, 10, 167–180, https://doi.org/10.5194/cp-10-
167-2014, 2014. Hunter, S. J., Haywood, A. M., Dolan, A. M., and Tindall, J. C.:
The HadCM3 contribution to PlioMIP phase 2, Clim. Past, 15,
1691–1713, https://doi.org/10.5194/cp-15-1691-2019, 2019. Raymo, M. E., Grant, B., Horowitz, M., and Rau, G. H.: Mid-
Pliocene warmth: stronger greenhouse and stronger conveyor,
Mar. Micropaleontol., 27, 313–326, 1996. Kamae, Y. and Ueda, H.: Mid-Pliocene global climate simula-
tion with MRI-CGCM2.3: set-up and initial results of PlioMIP
Experiments 1 and 2, Geosci. Model Dev., 5, 793–808,
https://doi.org/10.5194/gmd-5-793-2012, 2012. Rayner, N. A., Parker, D. E., Horton, E. B., Folland, C. K., Alexan-
der, L. V., Rowell, D. P., Kent, E. C., and Kaplan, A.: Global
analyses of sea surface temperature, sea ice, and night marine air
temperature since the late nineteenth century, J. Geophys. Res.,
108, 4407, https://doi.org/10.1029/2002JD002670, 2003 Kamae, Y., Yoshida, K., and Ueda, H.: Sensitivity of Pliocene cli-
mate simulations in MRI-CGCM2.3 to respective boundary con-
ditions, Clim. Past, 12, 1619–1634, https://doi.org/10.5194/cp-
12-1619-2016, 2016. Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 M.: Evaluation of Arctic warming
in mid-Pliocene climate simulations, Clim. Past, 16, 2325–2341,
https://doi.org/10.5194/cp-16-2325-2020, 2020. Haywood, A. M., Hill, D. J., Dolan, A. M., Otto-Bliesner, B. L.,
Bragg, F., Chan, W.-L., Chandler, M. A., Contoux, C., Dowsett,
H. J., Jost, A., Kamae, Y., Lohmann, G., Lunt, D. J., Abe-Ouchi,
A., Pickering, S. J., Ramstein, G., Rosenbloom, N. A., Salz-
mann, U., Sohl, L., Stepanek, C., Ueda, H., Yan, Q., and Zhang,
Z.: Large-scale features of Pliocene climate: results from the
Pliocene Model Intercomparison Project, Clim. Past, 9, 191–209,
https://doi.org/10.5194/cp-9-191-2013, 2013. Dowsett, H., Robinson, M., Haywood, A., Salzmann, U., Hill, D.,
Sohl, L., Chandler, M., Williams, M., Foley, K., and Stoll, D.:
The PRISM3D paleoenvironmental reconstruction, Stratigraphy,
7, 123–139, 2010. Dowsett, H., Dolan, A., Rowley, D., Moucha, R., Forte, A. M.,
Mitrovica, J. X., Pound, M., Salzmann, U., Robinson, M., Chan-
dler, M., Foley, K., and Haywood, A.: The PRISM4 (mid-
Piacenzian) paleoenvironmental reconstruction, Clim. Past, 12,
1519–1538, https://doi.org/10.5194/cp-12-1519-2016, 2016. Haywood, A. M., Dowsett, H. J., Dolan, A. M., Rowley, D.,
Abe-Ouchi, A., Otto-Bliesner, B., Chandler, M. A., Hunter, S. J., Lunt, D. J., Pound, M., and Salzmann, U.: The Pliocene
Model Intercomparison Project (PlioMIP) Phase 2: scientific
objectives and experimental design, Clim. Past, 12, 663–675,
https://doi.org/10.5194/cp-12-663-2016, 2016a. Dowsett, H. J., Cronin, T. M., Poore, R. Z., Thompson, R. S., What-
ley, R. C., and Wood, A. M.: Micropaleontological evidence for
increased meridional heat transport in the North Atlantic Ocean
during the Pliocene, Science, 258, 1133–1135, 1992. Haywood, A. M., Dowsett, H. J., and Dolan, A. M.: Inte-
grating
geological
archives
and
climate
models
for
the
mid-Pliocene
warm
period,
Nat. Commun.,
7,
10646,
https://doi.org/10.1038/ncomms10646, 2016b. Dowsett, H. J., Robinson, M. M., Haywood, A. M., Hill, D. J.,
Dolan, A. M., Stoll, D. K., Chan, W.-L., Abe-Ouchi, A., Chan-
dler, M. A., Rosenbloom, N. A., Otto-Bliesner, B. L., Bragg, F. Haywood, A. M., Tindall, J. C., Dowsett, H. J., Dolan, A. M., Fo-
ley, K. M., Hunter, S. J., Hill, D. J., Chan, W.-L., Abe-Ouchi, Clim. Past, 17, 529–543, 2021 https://doi.org/10.5194/cp-17-529-2021 Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 542 A., Stepanek, C., Lohmann, G., Chandan, D., Peltier, W. R., Tan,
N., Contoux, C., Ramstein, G., Li, X., Zhang, Z., Guo, C., Ni-
sancioglu, K. H., Zhang, Q., Li, Q., Kamae, Y., Chandler, M. Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 Rosenbloom, N. A., Otto-Bliesner, B. L., Brady, E. C., and
Lawrence, P. J.: Simulating the mid-Pliocene Warm Period
with the CCSM4 model, Geosci. Model Dev., 6, 549–561,
https://doi.org/10.5194/gmd-6-549-2013, 2013. Kanzow, T., Cunningham, S. A., Johns, W. E., Hirschi, J.J-M.,
Marotzke, J., Baringer, M. O., Meinen, C. S., Chidichimo, M. P., Atkinson, C., Beal, L. M., Bryden, H. L., and Collins, J.: Sea-
sonal Variability of the Atlantic Meridional Overturning Circula-
tion at 26.5◦N, J. Climate, 23, 5678–5698, 2010. Salzmann, U., Dolan, A. M., Haywood, A. M., Chan, W.-L., Voss,
J., Hill, D. J., Abe-Ouchi, A., Otto-Bliesner, B., Bragg, F. J.,
Chandler, M. A., Contoux, C., Dowsett, H. J., Jost, A., Kamae,
Y., Lohmann, G., Lunt, D. J., Pickering, S. J., Pound, M. J.,
Ramstein, G., Rosenbloom, N. A., Sohl, L., Stepanek, C., Ueda,
H., and Zhang, Z.: Challenges in quantifying Pliocene terrestrial
warming revealed by data–model discord, Nat. Clim. Change, 3,
969–974, 2013. Leduc, G., Garbe-Schönberg, D., Regenberg, M., Contoux, C.,
Etourneau, J., and Schneider, R.: The late Pliocene Benguela up-
welling status revisited by means of multiple temperature prox-
ies, Geochem. Geophy. Geosy., 15, 475–491, 2014. Small, R. J., Curchitser, E., Hedstrom, K., Kauffman, B., and
Large, W. G.: The Benguela Upwelling System: Quantifying
the Sensitivity to Resolution and Coastal Wind Representa-
tion in a Global Climate Model, J. Climate, 28, 9409–9432,
https://doi.org/10.1175/JCLI-D-15-0192.1, 2015. Li, X., Guo, C., Zhang, Z., Otterå, O. H., and Zhang, R.: PlioMIP2
simulations with NorESM-L and NorESM1-F, Clim. Past, 16,
183–197, https://doi.org/10.5194/cp-16-183-2020, 2020. Li, Z., Luo, Y., Arnold, N., and Tziperman, E.: Reductions in Strong
Upwelling-Favorable Wind Events in the Pliocene, Paleoceanog-
raphy and Paleoclimatology, 34, 1931–1944, 2019. Song, Z., Latif, M., Park, W., and Zhang, Y.: Influence of model
bias on simulating North Atlantic sea surface temperature during
the mid-Pliocene, Paleoceanography and Paleoclimatology, 33,
884–893, https://doi.org/10.1029/2018PA003397, 2018. Lurton, T., Balkkanski, Y., Bastrikov, V., Bekki, S., Bopp, L., Bra-
connot, P., Brockmann, P., Cadule, P., Contoux, C., Cozic, A.,
Cugnet, D., Dufresne, J.-L., Ethé, C., Foujols, M.-A., Ghattas, J.,
Hauglustaine, D., Hu, R.-M., Kageyama, M., Khodri, M., Lebas,
N., Levavasseur, G., Marchand, M., Ottlé, C., Peylin, P., Sima,
A. Szopa, S., Thiéblemont, R., Vuichard, N., and Boucher, O.:
Implementation of the CMIP6 forcing data in the IPSL-CM6A-
LR model, J. Adv. Model. Earth Sy., 12, e2019MS001940,
https://doi.org/10.1029/2019MS001940, 2020. Stepanek, C. and Lohmann, G.: Modelling mid-Pliocene cli-
mate with COSMOS, Geosci. https://doi.org/10.5194/cp-17-529-2021 Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 Model Dev., 5, 1221–1243,
https://doi.org/10.5194/gmd-5-1221-2012, 2012. Stepanek, C., Samakinwa, E., Knorr, G., and Lohmann, G.: Con-
tribution of the coupled atmosphere–ocean–sea ice–vegetation
model COSMOS to the PlioMIP2, Clim. Past, 16, 2275–2323,
https://doi.org/10.5194/cp-16-2275-2020, 2020. McClymont, E. L., Ford, H. L., Ho, S. L., Tindall, J. C., Haywood,
A. M., Alonso-Garcia, M., Bailey, I., Berke, M. A., Littler, K.,
Patterson, M. O., Petrick, B., Peterse, F., Ravelo, A. C., Rise-
brobakken, B., De Schepper, S., Swann, G. E. A., Thirumalai, K.,
Tierney, J. E., van der Weijst, C., White, S., Abe-Ouchi, A., Baat-
sen, M. L. J., Brady, E. C., Chan, W.-L., Chandan, D., Feng, R.,
Guo, C., von der Heydt, A. S., Hunter, S., Li, X., Lohmann, G.,
Nisancioglu, K. H., Otto-Bliesner, B. L., Peltier, W. R., Stepanek,
C., and Zhang, Z.: Lessons from a high-CO2 world: an ocean Tan, N., Contoux, C., Ramstein, G., Sun, Y., Dumas, C., Sepulchre,
P., and Guo, Z.: Modeling a modern-like pCO2 warm period
(Marine Isotope Stage KM5c) with two versions of an Institut
Pierre Simon Laplace atmosphere–ocean coupled general circu-
lation model, Clim. Past, 16, 1–16, https://doi.org/10.5194/cp-
16-1-2020, 2020. Williams, R. G., Roussenov, V., Lozier, M. S., and Smith, D.: Mech-
anisms of Heat Content and Thermocline Change in the Sub- Clim. Past, 17, 529–543, 2021 https://doi.org/10.5194/cp-17-529-2021 Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 Z. Zhang et al.: Mid-Pliocene Atlantic Meridional Overturning Circulation simulated in PlioMIP2 543 tropical and Subpolar North Atlantic, J. Climate, 28, 9803–9815,
2015. tropical and Subpolar North Atlantic, J. Climate, 28, 9803–9815,
2015. Zhang, Z.-S., Nisancioglu, K. H., Chandler, M. A., Haywood, A. M., Otto-Bliesner, B. L., Ramstein, G., Stepanek, C., Abe-Ouchi,
A., Chan, W.-L., Bragg, F. J., Contoux, C., Dolan, A. M., Hill, D. J., Jost, A., Kamae, Y., Lohmann, G., Lunt, D. J., Rosenbloom,
N. A., Sohl, L. E., and Ueda, H.: Mid-pliocene Atlantic Merid-
ional Overturning Circulation not unlike modern, Clim. Past, 9,
1495–1504, https://doi.org/10.5194/cp-9-1495-2013, 2013a. Zhang, Q., Li, Q., Zhang, Q., Berntell, E., Axelsson, J., Chen,
J., Han, Z., de Nooijer, W., Lu, Z., Wyser, K., and Yang,
S.: Simulating the mid-Holocene, Last Interglacial and mid-
Pliocene climate with EC-Earth3-LR, Geosci. Model Dev. Dis-
cuss. [preprint], https://doi.org/10.5194/gmd-2020-208, in re-
view, 2020. Zhang,
Z.-S.,
Nisancioglu,
K. H.,
and
Ninnemann,
U. S.:
Increased
ventilation
of
Antarctic
deep
water
dur-
ing
the
warm
mid-Pliocene,
Nat. Commun.,
4,
1499,
https://doi.org/10.1038/ncomms2521, 2013b. Zhang, Z. S., Nisancioglu, K., Bentsen, M., Tjiputra, J., Bethke,
I., Yan, Q., Risebrobakken, B., Andersson, C., and Jansen, E.:
Pre-industrial and mid-Pliocene simulations with NorESM-L,
Geosci. Model Dev., 5, 523–533, https://doi.org/10.5194/gmd-5-
523-2012, 2012. Zubakov, V. A. and Borzenkova, I. I.: Pliocene palaeoclimates: Past
climates as possible analogues of mid-twenty-first century cli-
mate, Palaeogeogr. Palaeocl., 65, 35–49, 1988. Clim. Past, 17, 529–543, 2021 https://doi.org/10.5194/cp-17-529-2021
|
https://openalex.org/W2912419107
|
https://www.journals.vu.lt/Bibliotheca-Lituana/article/download/12135/10720
|
Lithuanian
| null |
Krekenavos fotografo Tado Bajorūno darbo užrašai
|
Bibliotheca Lituana
| 2,018
|
cc-by
| 4,676
|
ISSN 2424-3477
ISBN 978-9955-33-708-9
Bibliotheca Lituana IV: privačioji raštija ir egodokumentinis
paveldas / Private Writings and Egodocumentary Heritage
2017, p. 209–222 1
LOČERIS, Petras. Atsiminimai. Užrašyti 1966–1971 m. Vilniuje. Biržų krašto muziejus „Sėla“,
BKM20860/51–75. Zita Pikelytė Panevėžio kraštotyros muziejus, Vasario 16-osios g. 23, Panevėžys. El. paštas zitapikelyte@gmail.com Straipsnyje nagrinėjami Panevėžio apskrities Krekenavos miestelyje
1920–1945 metais gyvenusio ir dirbusio fotografo Tado Bajorūno už
rašai – paprasta užrašų knygelė, kurioje surašytos jo 1934–1941 metais
gautos pajamos. Atlikę atvejo studiją – išanalizavę šiuos užrašus, galime
sužinoti, ar tuo laikotarpiu fotografija mažame miestelyje buvo pelnin
gas verslas ir kokia veikla, neskaitant fotografijos, fotografas dar užsiėmė,
kiek daugiausia pajamų gavo ir už kokią veiklą. Apibendrinę rezultatus
ir pasitelkę duomenis, atspindinčius kitų Lietuvos regionuose aptaria
muoju laikotarpiu dirbusių fotografų veiklą, galėsime susidaryti bendrą
jų ekonominės situacijos vaizdą. reikšminiai žodžiai: fotografas, Tadas Bajorūnas, pajamos, darbo
užrašai. P
rieškario fotografų darbo užrašų ar jų rašytų atsiminimų nėra labai
daug, tad tiriant to laikotarpio fotografiją kiekvienas autentiškas do
kumentas, atskleidžiantis šio amato atstovų darbo specifiką, veiklos sritis,
socialinę bei ekonominę padėtį, yra vertingas liudijimas. Bene didžiausius
atsiminimus apie savo gyvenimą ir fotografo kelią, pluoštelį sukurtų eilė
raščių yra palikęs biržietis Petras Ločeris – juos, fotografo ranka surašytus
į dvidešimt penkis mokyklinius sąsiuvinius, galime rasti ir patyrinėti Biržų
krašto muziejuje Sėla1. Surinktų atsiminimų apie vieno ar kito krašto foto
grafus saugoma ir kituose regionų muziejuose (Kretingos, Marijampolės P https://doi.org/10.15388/BibLita.2017.12135 209
Bibliotheca Lituana IV / 2017 Tadas Bajorūnas su žmona Kazimiera ir sūnumi
Petru. Apie 1926 m. T. Bajorūno nuotrauka kraštotyros, Mažeikių krašto, Že
maičių muziejuje Alka Telšiuose,
taip pat Vilniaus universiteto bi
bliotekos Rankraščių skyriuje ir
kt.), tačiau taip kruopščiai suda
rytus pajamų užrašus teko matyti
pirmą kartą. Tad šio tyrimo tiks
las – patyrinėti juos detaliau. Bibliotheca Lituana IV / 2017
210 Apie fotografą Tadas Bajorūnas gimė 1890 m. spalio 28 d. Navapolio dvare (ne
toli Sidabravo), ūkininkų Pauli
nos Pronis ir Justino Bajorūno
šeimoje. Tėvai valdė apie 80 hek
tarų žemės, užaugino 8 vaikus. Beveik visi vaikai pasekė tėvų
pėdomis: tapo žemdirbiais, tik
vienas išmoko kalvystės, o Tadas
– fotografo amato. 1919 m. Tadas vedė Kazimierą Jeleniauskaitę, Mikali
nos Riomerytės ir Petro Jeleniausko dukrą. Apie 1920 m. jauna šeima ap
sigyveno Krekenavoje, 29-ame Bažnyčios gatvės name. Jo gale fotografas
įsirengė ateljė su dideliais įstiklintais langais iš viršaus ir iš šono, kad gautų
natūralų dienos apšvietimą. Tadas Bajorūnas su žmona Kazimiera ir sūnumi
Petru. Apie 1926 m. T. Bajorūno nuotrauka Tadas Bajorūnas su žmona Kazimiera ir sūnumi
Petru. Apie 1926 m. T. Bajorūno nuotrauka Teigiama, jog fotografuoti T. Bajorūnas išmoko tarnaudamas carinės
Rusijos kariuomenėje. Buvo gabus, technikos dalykais itin besidomintis
žmogus, išmoko ne tik fotografuoti bei retušuoti negatyvus ir pozityvus,
bet ir taisyti laikrodžius, siuvimo mašinas, dviračius, radijo imtuvus, įvai
rius prietaisus. Būdamas didelis radijo mėgėjas, susikonstravo detektorinį
imtuvą ir nuolat klausėsi žinių, domėjosi įvykiais Lietuvoje ir pasaulyje. Ne vieną sukonstruotą radijo imtuvą yra ir pardavęs. Gyvenime T. Bajorūnas buvo linksmo būdo, turėjo iškalbos dovaną,
nuolat pokštaudavo, pasakodavo įvairias istorijas, mėgo žaisti šachmatais. Bibliotheca Lituana IV / 2017
210 ISSN 2424-3477, ISsBN 978-9955-33-708-9 Abu su žmona muzikavo: jis grojo mandolina, gražiai dainavo, o žmona
giedojo bažnyčios chore, skambino kanklėmis, 1924 m. net dalyvavo pir
moje dainų šventėje Kaune2. Tado Bajorūno darbo užrašai Tai nedidukė užrašų knygelė kietu viršeliu, rašyta pieštuku, laikui bėgant
apsitrynusi ir apdriskusi3. Ją iki šių dienų išsaugojo fotografo sūnus Vy
tautas, o 2012 m. kartu su krūva tėvo stiklo negatyvų patikėjo saugoti Pa
nevėžio kraštotyros muziejui. Knygelėje fotografo prirašyti 139 puslapiai. Pajamų apskaita pradėta vesti 1934 m., nuo vasario iki lapkričio mėnesio
uždarbio sumos ne visur beįskaitomos, aiškus tik bendras devynių mėnesių
(neskaitant sausio) uždarbis. Nuo 1934-ųjų lapkričio iki 1941 m. gegužės
mėnesio, išskyrus 1936 m. kovą ir maždaug nuo 1937 m. kovo vidurio iki pat
metų pabaigos, kai užrašai dėl ligos ar kitų priežasčių nebuvo pildomi, gana
smulkiai surašytos visos pajamos – už ką ir kiek pinigų gauta. Perskaičiavus
pateiktas pajamas, gautos sumos ne visur sutampa su fotografo nurodytais
skaičiais, kuriuos randame puslapio apačioje ar sudėtus mėnesio pabai
goje, vis dėlto, nors esant tam tikrai paklaidai, tyrimas leidžia pažvelgti į
anuometinę fotografo ekonominę padėtį ir veiklos sritis. Užrašai pildyti iki
1941 m. gegužės. Veikiausiai nuo to laiko T. Bajorūnas nebedirbo dėl ligos –
jis mirė 1945 m. kovo 13 d., kelerius metus prieš mirtį sunkiai sirgo4. 2
Plačiau: PIKELYTĖ, Zita. Miestelio fotografas ypač plušėjo per atlaidus. Panevėžio balsas,
2000, lapkričio 4, Nr. 255, p. 7; Tadas Bajarūnas ir jo fotografija. Sudarytoja ir teksto autorė
Zita Pikelytė. Panevėžys, 2000. 3
Užrašų knygelė saugoma Panevėžio kraštotyros muziejuje: PKM33064. 4
Iš autorės pokalbio su Vytautu Bajorūnu. ISSN 2424-3477, ISsBN 978-9955-33-708-9 3
Užrašų knygelė saugoma Panevėžio krašto
k lb Detali pajamų analizė Tado Bajorūno užrašuose pajamos smulkiai pradėtos fiksuoti nuo 1934 m. lapkričio mėnesio. Ne visus metus užrašai vesti 1936-aisiais – trūksta kovo
mėnesio duomenų, o 1937-aisiais pajamos pateiktos tik už sausio ir vasa
rio mėnesius, kovo pradžioje užrašai nutrūksta ir nepildomi iki pat metų
pabaigos. 1940 m. į valdžią atėjus sovietams, spalio 10 d. Lietuvos bankas Bibliotheca Lituana IV / 2017
211 ISSN 2424-3477, ISsBN 978-9955-33-708-9 buvo reorganizuotas į TSRS valstybinio banko Lietuvos Respublikos kon
torą, lapkričio 25 d. įvestas rublis, nuo 1941 m. kovo 25-osios uždrausta lito
apyvarta. Šalyje vykę pokyčiai atsispindi ir fotografo užrašuose. 1940 m. pabaigoje, kaip matyti, padidėjo įkainiai (nors iki pat metų pabaigos gau
tos pajamos dar nurodomos litais). Nuo 1941-ųjų apskaita jau vedama pa
jamas nurodant rubliais. Išanalizavus knygelės įrašus, pajamos išskirtos į tris pagrindines grupes
pagal veiklos rūšį, tai yra: už fotografijos paslaugas, už laikrodžių taisymą
ir už kitą veiklą gautos pajamos (už įvairius atliktus darbus ir kt.). 1 lente
lėje pateikiami duomenys atspindi, kokios buvo bendros T. Bajorūno metų
pajamos ir kokią pajamų dalį sudarė kiekviena iš šių grupių. Matyti, kad pagrindinis Tado Bajorūno pajamų šaltinis buvo fotogra
fija, kuri sudarė vidutiniškai 67 proc. visos užrašuose nurodytų pajamų
sumos. Patyrinėkime šią pajamų grupę detaliau. Įrašuose nuotraukas ga
lima suskirstyti į dvi pagrindines grupes pagal paskirtį, būtent: paso nuo
traukas ir atminčiai skirtas nuotraukas, fotografo vadintas fotonuotrau
komis, vestuvių, vaiko, pagrabo arba nabašninko nuotraukomis. Čia taip
pat priskaičiuotas ir atskirai įrašytas per Žolinės atlaidus gautas uždarbis. Užrašuose prie įrašo apie atliktą darbą dažniausia nurodomas ir kiekis,
pavyzdžiui, 2 paso nuotr. 5 Lt arba 6 fotonuotr. 30 Lt. Iš tokio įrašo galėtume
daryti išvadą, jog viena paso nuotrauka kainavo 2,5 lito, o viena nuotrauka
atminčiai – 5 litus. Kai kur prie įrašo fotonuotr. skaičius nenurodytas, o
įrašyta suma gana didelė, pavyzdžiui, 18 ar 24 litai. Kad galėtume padaryti tikslesnes išvadas dėl nuotraukų kainų, T. Bajo
rūno nurodytą gautą užmokestį palyginkime su kitų tada regionuose dir
busių fotografų taikytais įkainiais. Antai Mažeikių fotografui Hiršai Gur
vičiui už šešias 9×12 cm nuotraukas teko mokėti 3 litus, o už vieną didelę
18×24 cm nuotrauką – 4–5 litus, kitų Mažeikių fotografų teikiamos paslau
gos buvo vienu ar dviem litais pigesnės5. Biržietis Petras Ločeris už darbą
taip pat imdavo vienu ar dviem litais brangiau nei kiti Biržų fotografai: už
portretą 24×30 cm jam tekdavo mokėti 5–6 litus, už šešias 10×15 cm nuo
traukas – 6 litus. 5
Iš Mažeikių fotografijos istorijos (XIX a. pab.–XX a. I pusė): mašinraštis. Parengė Vytautas
Ramanauskas. Mažeikiai, 1992. Mažeikių muziejaus biblioteka, A-217 [b. p.]. p
7
DAUBARAITĖ-TESLENKIENĖ, Janina. Atsiminimai apie fotografą Juozą Daubarą. Iš Biržų
krašto fotografijos istorija: mašinraštis. Parengė Jonas Dagilis. Biržai, 1986. Biržų krašto mu
ziejus „Sėla“, MA29, b. 2, a. 4, p. 30–31. LOČERIS, Petras. Atsiminimai. BKM20860/73, p. 403, 405, 420–421. Detali pajamų analizė Jis dažnai buvo kviečiamas vykti į aplinkinius kaimus, kad Bibliotheca Lituana IV / 2017
212 ISSN 2424-3477, ISsBN 978-9955-33-708-9 ISSN 2424-3477, ISsBN 978-9955-33-708-9 1 lentelė. Tado Bajorūno užrašų knygelėje nurodytos pajamų rūšys
Metai
Bendros
metų
pajamos
Pajamos už
fotografijos
paslaugas
Pajamos už
laikrodžių
taisymą
Kitos
pajamos
1934 m. vasario–
gruodžio mėn. 2344,00 Lt
1934 m. lapkričio–
gruodžio mėn. 302,00 Lt
121,50 Lt
(40 %)
140,50 Lt
(47 %)
40,00 Lt
(13 %)
1935 m. 2495,50Lt
1740,00 Lt
(70 %)
589,50 Lt
(24 %)
166,00 Lt
(6 %)
1936 m. (išskyrus kovą)
2791,50 Lt
1896,00 Lt
(68 %)
590,50 Lt
(21 %)
305,00 Lt
(11 %)
1937 m. sausio–
vasario mėn. 422,50 Lt
312,00 Lt
(74 %)
82,50 Lt
(19 %)
28,00 Lt
(7 %)
1938 m. 2824,60 Lt
2166,80 Lt
(77 %)
495,50 Lt
(18 %)
162,30 Lt
(5 %)
1939 m. 2719,50 Lt
2064,50 Lt
(76 %)
480,00 Lt
(18 %)
175,00 Lt
(6 %)
1940 m. 4820,50 Lt
3748,50 Lt
(78 %)
823,00 Lt
(17 %)
249,00 Lt
(5 %)
1941 m. sausio–
balandžio mėn. 2357,00 rub. 1325,00 rub. (56 %)
820,00 rub. (35 %)
212,00 rub. (9 %) 1 lentelė. Tado Bajorūno užrašų knygelėje nurodytos pajamų rūšys įamžintų susirinkusią giminę, vestuves ar laidotuves. Atitinkamo formato
du fotografavimo seansai kainavo 50, 75 ar 100 litų. Savo atsiminimuose
P. Ločeris rašė, kad kainą nustatydavo atsižvelgdamas į socialinę kliento
padėtį (elgetas paso nuotraukoms fotografuodavo ir nemokamai (oficiali
jo nurodyta kaina buvo 2 litai, nesvarbu ar dvi, ar šešias nuotraukas kli
entas pageidavo gauti))6. Vabalninko fotografas Juozas Daubaras nurodo
tokius įkainius: stiklo negatyvo kaina – 2 litai ir kiekviena nuotrauka po
0,5 lito7. Tryškiuose fotografavęs Kazimieras Budginas už 6 nuotraukas ISSN 2424-3477, ISsBN 978-9955-33-708-9 Bibliotheca Lituana IV / 2017
213 2 lentelė. Tado Bajorūno užrašuose nurodytos fotografijų rūšys ir pajamos už jas 2 lentelė. Tado Bajorūno užrašuose nurodytos fotografijų rūšys ir pajamos už jas
Metai
Nuotraukų
atminčiai užsa
kymų skaičius /
gautos pajamos
Fotografuotų
vestuvių skai
čius / gautos
pajamos
Fotografuotų
laidotuvių
skaičius / gau
tos pajamos
Paso nuotrau
kų užsakymų
skaičius / gau
tos pajamos
1934 m. lapkričio –
gruodžio mėn. 13 / 64
1 / 5
1 / 5
23 / 47,5
1935 m. 218 / 924
9 / 42
13 / 68
331 / 692
1936 m. (išskyrus kovą)
333 / 1432,5
1 / 4
–
218 / 452,5
1937 m. sausio –
vasario mėn. 8
ŠLIVINSKAS, Almantas. Kuklūs žemaičiai, palikę pėdsaką Lietuvos fotografijos istorijoje.
Kalvotoji Žemaitija, 2004, liepos 17, Nr. 78, p. 5. ISSN 2424-3477, ISsBN 978-9955-33-708-9 Detali pajamų analizė 54 / 227
–
–
41 / 85
1938 m. 319 / 1390,5
–
–
356 / 739,8
1939 m. 305 / 1381,5
3 / 29
–
328 / 667
1940 m. 342 / 2051
11 / 108,5
5 / 27
523 / 1562
1941 m. sausio –
balandžio mėn. 47 / 576 rub. 2 / 30 rub. –
97 / 690 rub. (9×14 cm) prašydavo maždaug 3 litų8. Remdamiesi pateiktais duomeni
mis, galime daryti išvadą, jog T. Bajorūno užrašuose prie nuotraukų pa
rašyti skaičiai reiškia ne fotografijų, o užsakymų kiekį. Kiek užsakymų jis
per metus gaudavo ir kokios rūšies fotografijų turėjo padaryti, atsispindi 2
lentelėje. Čia iš nuotraukų atminčiai grupės į atskiras grafas išskirti vestu
vių ir laidotuvių nuotraukų užsakymai. Kaip matyti, daugiau pajamų gauta už atminčiai skirtas nuotraukas,
šiek tiek mažiau už paso nuotraukas. Tačiau 1940-aisiais, pasikeitus poli
tinei situacijai, pakito ir fotografijos paslaugų pobūdis – gyventojams teko
keisti asmens dokumentus, tad ypač padaugėjo užsakymų padaryti paso
nuotraukų. T. Bajorūno įkainiai buvo tokie: paso nuotraukos vidutiniškai
kainavo 2–2,5 lito, nuotraukų atminčiai kainos skyrėsi priklausomai nuo Bibliotheca Lituana IV / 2017
214 ISSN 2424-3477, ISsBN 978-9955-33-708-9 dydžio: už mažesnio formato9 nuotraukas dažniausia prašyta 4–5 litų, už
13×18 cm dydžio – 12 litų, kai kur nurodyta 17 litų, net 24 litai, vaikų nuo
traukos įkainotos 12–12,5 lito. Paprastai klientui būdavo padaromi 6 vie
netai. Kaip minėta, 1940 m. pabaigoje ėmė kilti paslaugų kainos: už paso
nuotraukas kai kur jau teko mokėti po 5–7 litus, už nuotraukas atminčiai
– 8–15 litų, o 1941-aisiais maždaug tokie pat įkainiai nusistovėjo ir rubliais. Fotografas atskirai išskyrė kelerius metus per Žolinės atlaidus gautas pa
jamas: 1935 m. atlikta 18 užsakymų už 76 litus, 1938 m. – 21 užsakymas už
84 litus, 1939 m. – 27 užsakymai už 108 litus. Daugiausia pajamų tiek už
fotografiją, tiek apskritai gauta 1938 metais. i
Kita stambi T. Bajorūno pajamų grupė – uždarbis už laikrodžių tai
symą. Detaliau šios pajamos dar išskirstytos pagal laikrodžių tipą, t. y. už
laikrodžių, žadintuvų, laikrodukų ir sieninių laikrodžių taisymą. Iš užrašų
matyti, kad laikrodžio taisymas kainavo 1–8 litus (didžiausia suma prie
žodžio laikrodis – 15 litų ir 20 litų), sieninio laikrodžio taisymas – 4–8 litus,
žadintuvo, užrašuose vadinamo budilneku, – dažniausia 1,5–2,5 ar 3 litus;
1941 m. už laikrodžio taisymą dažniausia tekdavo mokėti 7–20 rublių (di
džiausia suma – 30 rublių), už žadintuvo taisymą – 7–15 rublių. 9
T. Bajorūno užrašuose nuotraukų dydžiai dažniausia nenurodomi, Panevėžio kraštotyros
muziejuje saugomų jo nuotraukų dydis dažniausia 8,3×13,3 cm, 8,6×13,6 cm, 9×14 cm. 10 Panevėžio miesto turgaus kainų biuletenis. LCVA, f. 816, ap. 1, b. 17, l. 5, 16. Detali pajamų analizė Trečiai grupei priskirtos visos likusios pajamos, kurios sudaro mažiau
sią dalį. Tai užmokestis už įvairių prietaisų (patefonų, radijo imtuvų, au
sinių, žibalinių / benzininių viryklių (primusų), siuvimo mašinų, skėčių,
lempų ir kt.), akinių, žiedų, dviračių, motociklų taisymą. Įrašuose vartoja
ma buitinė kalba, tarmiški bei iš kitų kalbų pasiskolinti daiktų pavadini
mai, pavyzdžiui: ridekiulis, veizdeliai, broškie ir t. t. Yra kelios žymos radijas
arba pardaviau radiją, prie kurių nurodoma suma: 15, 19, 21, 25 ar 30 litų. Radijo imtuvų ir jų remonto darbų paklausa pastebima 1940 metais. Sudėję duomenis galime sužinoti, kokios maždaug buvo T. Bajorūno
mėnesio pajamos. 1934 m. (neskaitant sausio) per mėnesį jis vidutiniškai
uždirbo 213 litų, 1935 m. – 208 litus, 1936 m. (išskyrus kovą) – 254 litus,
1937 m. sausio–vasario mėnesiais – 211 litų, 1938 m. – 235 litus, 1939 m. –
227 litus, 1940 m. – 402 litus, 1941 m. sausio–balandžio mėnesiais – 589
rublius. Sudėjus visų 1935, 1938 ir 1939 metų bei 1936 metų 11-os mėnesių ISSN 2424-3477, ISsBN 978-9955-33-708-9 Bibliotheca Lituana IV / 2017
215 Tado Bajorūno užrašų 1935 metų vasario mėnesio įrašai Tado Bajorūno užrašų 1935 metų vasario mėnesio įrašai pajamas ir padaliję iš tuos metus dirbtų mėnesių skaičiaus, išeina, kad vi
dutinis T. Bajorūno mėnesio uždarbis per šiuos metus siekė 230 litų, iš jų
už fotografijos paslaugas per mėnesį vidutiniškai uždirbta po 164 litus. Ar toks uždarbis fotografui garantavo aukštą pragyvenimo lygį? Paž
velkime į 1934 metų kainas Panevėžio turguje: rugių centneris kainavo
10 litų, kviečių – 13,50 lito, avižų – 9 litus, bulvių – 3 litus; melžiama kar
vė – 100–180 litų, darbinis arklys – 100–150 litų, mėsinė avis – 8–12 litų,
gyvas kalakutas – 5 litus, gyva žąsis – 6 litus, gyva višta – 2 litus; litras pie
no – 0,25 lito, grietinės – 1,20 lito, kilogramas sviesto – 3 litus, bičių medaus
– 3 litus, sėmenų aliejaus – 1,50 lito, 10 vienetų didelių kiaušinių– 1,70 lito,
mažų – 1,50 lito; vyriškas vilnonis kostiumas – 100 litų, medvilninė sukne
lė – 10 litų10. Remiantis nuo 1924 m. gegužės 1 d. įsigaliojusiu Valstybės tar
nautojų atlyginimo įstatymo II priedu, žemiausios (I) kategorijos I laipsnio
tarnautojo mėnesio alga buvo 150 litų, aukščiausios (XVIII) kategorijos – Bibliotheca Lituana IV / 2017
216 ISSN 2424-3477, ISsBN 978-9955-33-708-9 ISSN 2424-3477, ISsBN 978-9955-33-708-9 1200 litų11. Detali pajamų analizė Amžininkų atsiminimuose rašoma, kad kraštotyrininkas ir fo
tografas Balys Buračas, 1931–1933 metais dirbdamas Šiaulių „Aušros“ mu
ziejuje, gaudavo apie 100–130 litų mėnesio atlyginimą12, taigi jo atlygis buvo
keliomis dešimtimis mažesnis, o T. Bajorūno uždarbis už fotografiją vos
keliolika litų didesnis už nustatytą minimalią tarnautojo algą. 1
Valstybės tarnautojų atlyginimo įstatymo II priedas. Vyriausybės žinios, 1924, Nr. 165, p. 2.i 12 Vinco Vaitekūno prisiminimai apie Balį Buračą. Parengė Birutė Kulnytė. Etnografija, Nr. 1,
p. 45–46.i 3 Žr. PIKELYTĖ, Zita. Lietuvos provincijos fotografija 1918–1940 metais: daktaro disertacija. Vil
nius, 2012, p. 70.i 12 Vinco Vaitekūno prisiminimai apie Balį Buračą. Parengė Birutė Kulnytė. Etnografija, Nr. 1,
p. 45–46.
13 Žr. PIKELYTĖ, Zita. Lietuvos provincijos fotografija 1918–1940 metais: daktaro disertacija. Vil
nius, 2012, p. 70.i 1927 m., saugoma Panevėžio kraštotyros muziejuje: PKM19409, F4252.
15 GIRININKIENĖ, Vida. Fotografas Jonas Sinkevičius. Iš Papilė, 1 d. Sudarytojai Vida Giri
ninkienė, Leopoldas Rozga, Rita Regina Trimonienė, Povilas Krikščiūnas, Juozas Pabrėža.
Vilnius, 2004, p. 704–705. Bibliotheca Lituana IV / 2017
217 11
Valstybės tarnautojų atlyginimo įstatymo II priedas. Vyriausybės žinios, 1924, Nr. 165, p. 2.
12 Vinco Vaitekūno prisiminimai apie Balį Buračą. Parengė Birutė Kulnytė. Etnografija, Nr. 1,
p. 45–46.
13 Žr. PIKELYTĖ, Zita. Lietuvos provincijos fotografija 1918–1940 metais: daktaro disertacija. Vil
nius, 2012, p. 70.
14 Tokiu antspaudu pažymėta M. Gudzevičiaus fotografija Vilkijos progimnazijos moksleiviai.
1927 m., saugoma Panevėžio kraštotyros muziejuje: PKM19409, F4252.
15 GIRININKIENĖ, Vida. Fotografas Jonas Sinkevičius. Iš Papilė, 1 d. Sudarytojai Vida Giri
ninkienė, Leopoldas Rozga, Rita Regina Trimonienė, Povilas Krikščiūnas, Juozas Pabrėža.
Vilnius, 2004, p. 704–705. 4 Tokiu antspaudu pažymėta M. Gudzevičiaus fotografija Vilkijos progimnazijos moksleivia
1927 m., saugoma Panevėžio kraštotyros muziejuje: PKM19409, F4252. Papildomas miestelių fotografų verslas Ne tik T. Bajorūnas, bet ir kiti mažesnių miestelių fotografai turėjo po kelis
amatus, vertėsi papildoma veikla. Daugelis iš jų buvo sumanūs eksperi
mentuotojai, konstruotojai, remontininkai, gebėjo atlikti didelio kruopš
tumo, kantrybės ir tikslumo reikalaujančius darbus. Kai kurie iš jų, tokie
kaip Romas Paliulionis iš Apasčios ar Martynas Kavolis iš Dieglių kaimo,
net pirmuosius fotoaparatus susikonstravo patys13. Vienas iš dažnesnių papildomų užsiėmimų, kaip matyti ir iš Tado Ba
jorūno pajamų analizės, buvo laikrodininko amatas. Kartais net antspau
duose, kuriuos dėdavo ant nuotraukų, buvo nurodomi abu amatai. Tokį
antspaudą turėjo Vilkijos fotografas ir laikrodininkas Martynas Gudzevi
čius14. Laikrodžius taisyti mokėjo ir Juozas Daubaras, Jonas Sinkevičius –
pastarojo tėvas laikrodininko ir juvelyro profesiją įgijo Šveicarijoje, žinias
ir įgūdžius perdavė visiems savo sūnums: dviem iš jų tai tapo kasdiene
duona, o trečiasis – Jonas – tapo dar ir žymiu Papilės fotografu15. O pas
Viekšnių fotografą Joną Kinčiną, kaip pas gerai žinomą laikrodininką,
klientų atvykdavo net iš Kauno. Prie jo namų greta viena kitos stovėjo
dvi reklaminės lentos: FOTOGRAFIJA J. KINČINO ir SUTAISO LAIKRO
DŽIUS J. KINČINAS. Teigiama, kad jis buvo ne šiaip sau laikrodininkas: Bibliotheca Lituana IV / 2017
217 ISSN 2424-3477, ISsBN 978-9955-33-708-9 turėjo frezavimo mašinėlę, kuria išfrezuodavo bet kokias ašeles ir ratukus,
pats gamino laikrodžių rodykles, kišeninių bei rankinių laikrodžių stikliu
kus, turėjo mufelinę krosnį, kurioje lydė stiklą16. J. Kinčinas išbandė ir daugiau įvairių verslo rūšių. Apie 1925–1930 m. jis
užsiėmė dviračių nuoma, bet jie dažnai lūždavo, tad nemažai laiko tekdavo
skirti remontui. Tada pradėjo gaminti ledus: žiemą pasamdydavo žmo
nių, kad jie nuo užšalusios Ventos upės pripjaustytų ledų, sukraudavo juos
daržinėje ir apipildavo pjuvenomis. Taip ledai išsilaikydavo visą vasarą. Bet ir šis verslas truko neilgai, nes teko samdyti daug darbininkų priruoš
ti ledų ir realizuoti juos. Vėliau jis lankė keramikos kursus, kad išmoktų
gaminti nuotraukas antkapiams17. Pas J. Daubarą buvo galima ne tik nu
sifotografuoti, bet ir apsikirpti, išsinuomoti dviratį; būdamas raštingas ir
mokėdamas daug kalbų (anglų, vokiečių, rusų, lenkų, jidiš, latvių), reika
lui esant, jis parašydavo reikiamą raštą, jaunimo vakarėliuose už pinigus
pagrieždavo armonika ar akordeonu18. Pirmiausia Subačiaus fotografas Povilas Šinskis išmoko siuvėjo amato. Įgudęs fotografuoti jis vertėsi abiem amatais ir tik pasistatęs namus bei įsi
rengęs fotopaviljoną ėmė daugiau užsiimti fotografija19. Ne tik fotografuo
damas, bet ir siūdamas bei karšdamas vilnas užsidirbdavo Antanas Mic
kus iš Veliuonos. Ant Mickų namo sienos kabojo reklama: A. MICKAUS
VILNŲ KARŠTUVAI IR FOTOGRAFIJA20. 1934 m. 16 PLASTININA, Bernarda. Į fotoaparato objektyvą sutilpo visi Viekšniai. Santarvė, 2004, ge
gužės 6, Nr. 51, p. 6; ŽVIRGŽDAS, Stanislovas. Mūsų miestelių fotografai. Vilnius, 2003, p. 129.
17 PLASTININA, Bernarda. Į fotoaparato objektyvą sutilpo visi Viekšniai. Santarvė, 2004, ge
gužės 27, Nr. 60, p. 2.
18 ŽVIRGŽDAS, Stanislovas. Mūsų miestelių fotografai, p. 5.
19 Žr. PIKELYTĖ, Zita. Vaizdais prakalbusi praeitis. Iš ŠINSKIS, Povilas. Fotografija. Panevėžys,
2005, p. 4–5.
20 GIRININKIENĖ, Vida. Veliuoniečių kasdienybė Antano Mickaus nuotraukose (1925–1937).
Liaudies kultūra. 2001, Nr. 1, p. 48–53.
21 ŽVIRGŽDAS, Stanislovas. Mūsų miestelių fotografai, p. 65. ISSN 2424-3477, ISsBN 978-9955-33-708-9 22 Žr. PIKELYTĖ, Zita. Lietuvos provincijos fotografija 1918–1940 metais, p. 64–72.
23 GULMANAS, Viktoras. Iš fotografijos vystymosi Sūduvoje istorijos. 1839–1941 m.: mašinraštis.
Kapsukas, 1977. Marijampolės kraštotyros muziejus, MKM15088, R3765, p. 26.
24 Iš Mažeikių fotografijos istorijos (XIX a. pab.–XX a. I pusė), [b. p.]. ISSN 2424-3477, ISsBN 978-9955-33-708-9 25 Ateljė MENAS reklaminis skelbimas. Šiaurės Lietuva. 1932, sausio 3, Nr. 1, p. 7; 1932, sausio 10,
Nr. 2, p. 7; 1932, sausio 17, Nr. 3, p. 4; 1932, sausio 24, Nr.4, p. 6; 1932, sausio 31, Nr. 5, p. 6. Išvados Fotografo Tado Bajorūno darbo užrašai – tai ne tik duomenys apie 1934–
1941 metais gautas jo pajamas (kiek, už ką ir kurį mėnesį jis uždirbo). Jie
atskleidžia nemažai informacijos ir apie fotografo darbo pobūdį, raštingu
mą, kalbos vartosenos ypatumus. Šių užrašų analizė bei straipsnyje pateikiami duomenys apie kitų tuo lai
kotarpiu regionuose dirbusių fotografų veiklą, informacija apie 1934 metų
Panevėžio turgaus kainas leidžia susidaryti bendrą vaizdą, kokia buvo eko
nominė mažo Nepriklausomos Lietuvos miestelio fotografo situacija: pra
gyvenimo lygis, fotografijos paslaugų įkainiai, papildomas verslas. 1940–1941 metų įrašai rodo, kokių pokyčių kasdieniame žmogaus gy
venime lėmė sovietinė okupacija ir artėjančio karo nuojauta: pakeista va
liuta, išaugo paslaugų kainos, atsirado didesnis paso nuotraukų poreikis,
padidėjo radijo imtuvų paklausa. Šie užrašai – tai unikalus dokumentas ir įdomus mūsų krašto praeities
pažinimo šaltinis. Papildomas miestelių fotografų verslas nagingas veliuoniškis
išradimui „Siuvamosios mašinos patobulinimas ypatingai storai me
džiagai siūti“ įsigijo patentą. Be to, jis gamino naminį vyšnių ir obuolių
vyną, tam tikslui net buvo nusipirkęs specialių prietaisų: priemonę vyno
stiprumui matuoti, sacharimetrą, keletą 60 litrų talpos indų21. Naminį
obuolių vyną gamino ir miežiškietis Vincas Firinauskas – jis labai mėgo Bibliotheca Lituana IV / 2017
218 ISSN 2424-3477, ISsBN 978-9955-33-708-9 dirbti sode, sodinti ir prižiūrėti vaismedžius, skiepyti juos, išvesti naujas
veisles. Obuolių turėjo parduoti ir vasarą, ir žiemą. Neskaitant fotografijos,
V. Firinauskas dar mokėjo dailidės amatą. Gamino jis baldus ir ūkio ra
kandus, pasižymėjo kruopštumu ir kantrumu, o jo gaminiai išsiskyrė itin
gera kokybe. Jis pats virė medžio klijus, savo gaminius, ypač baldus, mėgo
puošti įvairiomis iš knygų nužiūrėtomis ar paties sugalvotomis detalėmis,
ornamentais. V. Firinauskas, kaip ir prieniškis Jokūbas Skrinskas bei Jo
nas Kinčinas, buvo aistringi kino gerbėjai, pastarieji du net patys kurį laiką
rodė kiną. Papildomų pajamų fotografai gaudavo ir pardavinėdami savo
fotografuotų vaizdų atvirukus, už šalies laikraščiams ir žurnalams pateik
tas nuotraukas, Petras Ločeris mėgino įsitvirtinti fotoplokštelių gamybos
versle ir t. t.22. Mažuose miesteliuose dažniausia dirbo tik po vieną fotografą. Daugiau
gyventojų turinčiuose miestuose fotografijos paslaugų paklausa buvo di
desnė, tad stambesnių fotografijos įmonių savininkai į verslą įtraukdavo
šeimos narius, dažnai turėjo mokinių, samdomų darbuotojų. Pavyzdžiui,
Marijampolės fotografas Jakobas Vindsbergas turėjo padėjėją Jakobsoną,
kuriam mokėjo 250 litų mėnesio algą23. Mažeikietis Hirša Gurvičius už
septynių pozityvų (1 bandomojo ir 6 vienetų klientui) išspausdinimą įgu
dusiam mokiniui ar padėjėjui mokėdavo po vieną litą, o šis per dieną vi
dutiniškai padarydavo nuotraukų iš dešimties negatyvų24. Amato išmoku
siems mokiniams, mėginantiems įsitvirtinti fotografijos versle, tekdavo
susidurti su gana didele konkurencija, tad dalis jų taip ir likdavo dirbti pas
savo mokytojus retušuotojais, jei tik šie priimdavo. Net ir pasisekus verslui, norėdami išsilaikyti ir gauti užtektinai pelno,
fotografai turėjo siekti aukštos darbo kokybės, didinti siūlomų paslaugų
įvairovę, tinkamai jas reklamuoti. Jau ir ano meto laikraščiuose galima rasti
reklaminių skelbimų, kuriuose klientai vilioti akcijomis. Antai visą 1932 m. sausio mėnesį laikraštyje „Šiaurės Lietuva“ buvo spausdinamas ateljė Me
nas, veikusios Šiauliuose, Vilniaus g. 158, skelbimas, kad joje švenčių proga ISSN 2424-3477, ISsBN 978-9955-33-708-9 Bibliotheca Lituana IV / 2017
219 „nebuvusis fotografijų pigumas“, kur „pirmaklasinėj dail[ės] fotografijoj“
nusifotografuoti galima tiktai už vieną litą ir tiktai už tris litus galima gauti
„padidintą didelį dailės portretą“25 [paryškinta skelbime – Z. P.]. ISSN 2424-3477, ISsBN 978-9955-33-708-9 1. Apie Suvalkijos fotografus. Surinko Viktoras Gulmanas, 1977, Vilniaus universiteto bibliotekos
Rankraščių skyrius (toliau – VUB RS), F113–277.i Šaltiniai ir literatūra 1. Apie Suvalkijos fotografus. Surinko Viktoras Gulmanas, 1977, Vilniaus universiteto bibliotekos
Rankraščių skyrius (toliau – VUB RS), F113–277.i 2. Biržų krašto fotografijos istorija: mašinraštis. Parengė Jonas Dagilis. Biržai, 1986. Biržų krašto
muziejus Sėla (toliau – BKM), MA29, b. 2, a. 4. 2. Biržų krašto fotografijos istorija: mašinraštis. Parengė Jonas Dagilis. Biržai, 1986. Biržų krašto
muziejus Sėla (toliau – BKM), MA29, b. 2, a. 4. 3. DAUBARAITĖ-TESLENKIENĖ, Janina. Atsiminimai apie fotografą Juozą Daubarą. Iš Biržų kraš
to fotografijos istorija: mašinraštis. Parengė Jonas Dagilis. Biržai, 1986. BKM, MA29, p. 30–31. 3. DAUBARAITĖ-TESLENKIENĖ, Janina. Atsiminimai apie fotografą Juozą Daubarą. Iš Biržų kraš
to fotografijos istorija: mašinraštis. Parengė Jonas Dagilis. Biržai, 1986. BKM, MA29, p. 30–31. 4. GIRININKIENĖ, Vida. Fotografas Jonas Sinkevičius. Iš Papilė, 1 d. Sudarytojai Vida Giri
ninkienė, Leopoldas Rozga, Rita Regina Trimonienė, Povilas Krikščiūnas, Juozas Pabrėža. Vilnius: Versmė, 2004, p. 704–705. ISBN 9986-9236-8-9. 4. GIRININKIENĖ, Vida. Fotografas Jonas Sinkevičius. Iš Papilė, 1 d. Sudarytojai Vida Giri
ninkienė, Leopoldas Rozga, Rita Regina Trimonienė, Povilas Krikščiūnas, Juozas Pabrėža. Vilnius: Versmė, 2004, p. 704–705. ISBN 9986-9236-8-9. Bibliotheca Lituana IV / 2017
220 ISSN 2424-3477, ISsBN 978-9955-33-708-9 5. GIRININKIENĖ, Vida. Veliuoniečių kasdienybė Antano Mickaus nuotraukose (1925–1937). Liaudies kultūra, 2001, Nr. 1, p. 48–53. ISSN 0236-0551. 6. GULMANAS, Viktoras. Iš fotografijos vystymosi Sūduvoje istorijos. 1839–1941 m.: m
Kapsukas, 1977. Marijampolės kraštotyros muziejus, MKM15088, R3765, p. 26. 7. Iš Mažeikių fotografijos istorijos (XIX a. pab.–XX a. I pusė): mašinraštis. Parengė Vytautas
Ramanauskas. Mažeikiai, 1992, [b. p.]. Mažeikių muziejaus biblioteka, A-217. 8. LOČERIS, Petras. Atsiminimai. Užrašyti 1966–1971 m. Vilniuje. BKM20860/51–75;
BKM20860/73, p. 403, 405, 420–421. 9. Martyno Kavolio atsiminimai. Užrašė Virgilijus Juodakis, 1972. VUB RS, F113–189. 10. Panevėžio miesto turgaus kainų biuletenis. Lietuvos centrinis valstybės archyvas, f. 816, ap. 1, b. 17, l. 5, 16. 11. Petro Ločerio atsiminimai. Užrašė Tolvaišas. Vilnius, 1968. VUB RS, F113–184.i 12. PIKELYTĖ, Zita. Lietuvos provincijos fotografija 1918–1940 metais: daktaro disertacija. Vilnius:
VDA, 2012, p. 68–72. ISBN 978-609-447-058-5. 13. PIKELYTĖ, Zita. Miestelio fotografas ypač plušėjo per atlaidus. Panevėžio balsas, 2000,
lapkričio 4, Nr. 255, p. 7. ISSN 1392-8201.i 14. PIKELYTĖ, Zita. Vaizdais prakalbusi praeitis. Iš Povilas Šinskis. Fotografija. Panevėžys: Pane
vėžio kraštotyros muziejus, 2005, p. 4–5. ISBN 9955-9643-4-0. 15. PLASTININA, Bernarda. Į fotoaparato objektyvą sutilpo visi Viekšniai. Santarvė, 2004, gegu
žės 6, Nr. 51, p. 6. ISSN 1648-3286. 16. PLASTININA, Bernarda. Į fotoaparato objektyvą sutilpo visi Viekšniai. Santarvė, 2004, gegu
žės 27, Nr. 60, p. 2. ISSN 1648-3286. 17. Šiaurės Lietuva. ISSN 2424-3477, ISsBN 978-9955-33-708-9 Bibliotheca Lituana IV / 2017
221 Šaltiniai ir literatūra 1932, sausio 3, Nr. 1, p. 7; 1932, sausio 10, Nr. 2, p. 7; 1932, sausio 17, Nr. 3, p. 4;
1932, sausio 24, Nr.4, p. 6; 1932, sausio 31, Nr. 5, p. 6. ISSN 2029-8196. 18. ŠLIVINSKAS, Almantas. Kuklūs žemaičiai, palikę pėdsaką Lietuvos fotografijos istorijoje. Kalvotoji Žemaitija, 2004, liepos 17, Nr. 78, p. 5. ISSN 2029-1337. 19. Tadas Bajarūnas ir jo fotografija. Sudarytoja ir teksto autorė Zita Pikelytė. Panevėžys: Panevė
žio kraštotyros muziejus, 2000. 20. Tado Bajorūno darbo užrašų knygelė. Panevėžio kraštotyros muziejus, PKM33064. 21. Valstybės tarnautojų atlyginimo įstatymo II priedas. Vyriausybės žinios, 1924, Nr. 165, p. 2 22. Vinco Vaitekūno prisiminimai apie Balį Buračą. Parengė Birutė Kulnytė. Etnografija, Nr. 1,
Vilnius: Lietuvos istorijos ir etnografijos muziejus, 1991, p. 45–46. ISBN 1648-4835. 23. ŽVIRGŽDAS, Stanislovas. Mūsų miestelių fotografai. Vilnius: Lietuvos fotomenininkų sąjun
ga, 2003, p. 5, 65, 129. ISBN 9955-438-18-5. Bibliotheca Lituana IV / 2017
221 ISSN 2424-3477, ISsBN 978-9955-33-708-9 Bibliotheca Lituana IV / 2017
222 ISSN 2424-3477, ISsBN 978-9955-33-708-9 summaryh The article analyzes the records of the photographer Tadas Bajorūnas, who lived
and worked in Krekenava town (Panevėžys County) in 1920–1945. These records
are written in a simple notebook, which contains the photographer‘s generated
revenues of the year 1934–1941. After making a study of the notes, we can see if in
that period the photography taking was a profitable business in a small town. Also
we could see in what additional activities the photographer was engaged in and
what kind of activities were most profitable for him. t After the analysis of the notebook records the revenues were divided into three
main groups according to the type of activity: the revenue from photographic ser
vices (they accounted for an average of 67 per cent of all the income records),
the revenue from watch repairing and other revenue (for various jobs, etc.). Data
collected from various sources show that not just T. Bajorūnas, but also the other
photographers from smaller towns had a few crafts and were engaged in addition
al businesses. Many of them were skilled experimenters, constructers, repairers,
who were able to carry out works, which required large-diligence, patience and
precision. Tadas Bajorūnas’ records reveal not only his economic situation, but also a lot
of information about the nature of his work, literacy, language usage nuances. It is
the unique document and the interesting source of the knowledge of our country’s
past events.
|
https://openalex.org/W2767237799
|
https://europepmc.org/articles/pmc5713315?pdf=render
|
English
| null |
Modulation of Cell Death Pathways by Hepatitis C Virus Proteins in Huh7.5 Hepatoma Cells
|
International journal of molecular sciences
| 2,017
|
cc-by
| 12,973
|
Received: 20 September 2017; Accepted: 3 November 2017; Published: 6 November 2017 Received: 20 September 2017; Accepted: 3 November 2017; Published: 6 November 2017 Abstract: The hepatitis C virus (HCV) causes chronic liver disease leading to fibrosis, cirrhosis,
and hepatocellular carcinoma. HCV infection triggers various types of cell death which contribute
to hepatitis C pathogenesis. However, much is still unknown about the impact of viral proteins
on them. Here we present the results of simultaneous immunocytochemical analysis of markers
of apoptosis, autophagy, and necrosis in Huh7.5 cells expressing individual HCV proteins or their
combinations, or harboring the virus replicon. Stable replication of the full-length HCV genome
or transient expression of its core, E1/E2, NS3 and NS5B led to the death of 20–47% cells, 72 h
posttransfection, whereas the expression of the NS4A/B, NS5A or NS3-NS5B polyprotein did
not affect cell viability. HCV proteins caused different impacts on the activation of caspases-3,
-8 and -9 and on DNA fragmentation. The structural core and E1/E2 proteins promoted apoptosis,
whereas non-structural NS4A/B, NS5A, NS5B suppressed apoptosis by blocking various members
of the caspase cascade. The majority of HCV proteins also enhanced autophagy, while NS5A also
induced necrosis. As a result, the death of Huh7.5 cells expressing the HCV core was induced
via apoptosis, the cells expressing NS3 and NS5B via autophagy-associated death, and the cells
expressing E1/E2 glycoproteins or harboring HCV the replicon via both apoptosis and autophagy. Keywords: hepatitis C virus; hepatoma Huh7.5 cells; apoptosis; autophagy; necrosis; caspase; replicon Article
Modulation of Cell Death Pathways by Hepatitis C
Virus Proteins in Huh7.5 Hepatoma Cells Olga V. Masalova 1,*, Ekaterina I. Lesnova 1, Pavel N. Solyev 2
ID , Natalia F. Zakirova 2,
Vladimir S. Prassolov 2, Sergey N. Kochetkov 2, Alexander V. Ivanov 2,*
ID and Alla A. Kushch 1 1
Ivanovsky Institute of Virology, Gamaleya National Research Center of Epidemiology and Microbiology,
Ministry of Health of the Russian Federation, Moscow 123098, Russia; wolf252006@yandex.ru (E.I.L.);
vitallku@mail.ru (A.A.K.)
2 1
Ivanovsky Institute of Virology, Gamaleya National Research Center of Epidemiology and Microbiology,
Ministry of Health of the Russian Federation, Moscow 123098, Russia; wolf252006@yandex.ru (E.I.L.);
vitallku@mail.ru (A.A.K.) 2
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, Russia;
solyev@gmail.com (P.N.S.); nat_zakirova@mail.ru (N.F.Z.); prassolov45@mail.ru (V.S.P.);
kochet@eimb.ru (S.N.K.) 2
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, Russia;
solyev@gmail.com (P.N.S.); nat_zakirova@mail.ru (N.F.Z.); prassolov45@mail.ru (V.S.P.);
kochet@eimb.ru (S.N.K.) *
Correspondence: ol.mas@mail.ru (O.V.M.); aivanov@yandex.ru (A.V.I.); Tel.: +7-499-190-30-49 (O.V.M.);
+7-499-135-60-65 (A.V.I.) *
Correspondence: ol.mas@mail.ru (O.V.M.); aivanov@yandex.ru (A.V.I.); Tel.: +7-499-190-30-49 (O.V.M.);
+7-499-135-60-65 (A.V.I.) International Journal of
Molecular Sciences International Journal of
Molecular Sciences 1. Introduction The hepatitis C virus (HCV) is a wide-spread etiological agent that often leads to chronic liver
disease. Chronic hepatitis C (CHC) is strongly associated with the development of liver fibrosis,
cirrhosis, and hepatocellular carcinoma (HCC). The exact mechanisms of HCV pathogenesis are still
not completely understood. Numerous evidence suggest that pathogenic events are promoted by
hepatocyte death in various forms such as apoptosis, autophagy, and necrosis [1–3]. Apoptosis plays
a prominent role in maintaining tissue homeostasis by the removal of senescent cells which have
been damaged by various factors including viral infections, or by the removal of rapidly proliferating
transformed cells. The outer apoptotic pathway is mediated by the transmembrane death receptors
that belong to the tumor necrosis factor (TNF) family and by activation of the initiator caspase-8 [4]. The inner pathway is induced in response to DNA damage, pronounced oxidative stress, increase
of intracellular Ca2+ levels, ER stress and unfolded protein response (UPR) [4]. These events lead to
mitochondrial dysfunction that triggers activation of the initiation caspase-9 [4,5]. Both outer and Int. J. Mol. Sci. 2017, 18, 2346; doi:10.3390/ijms18112346 www.mdpi.com/journal/ijms 2 of 16 Int. J. Mol. Sci. 2017, 18, 2346 inner pathways activate the key effector caspase-3 [4]. The latter, in its turn, triggers the breakdown
of nucleus-activating DNAases that degrade DNA to nucleosome fragments. HCV infection was
earlier shown to induce both of these caspase-mediated apoptotic pathways. In particular, HCV
directly triggers unfolded protein response and calcium-dependent mitochondrial dysfunction [6]. Normally, apoptotic cell death is not accompanied by inflammation. However, persistent viral
infections and extensive production of pro-apoptotic stimulus (death receptors, inner signals such
as cytochrome c, Bax, Bad and Bak proteins) promote intensive and long-lasting inflammation that
can result in massive loss of hepatocytes, ischemic liver damage, and irreversible organ damage to
follow [7]. At the same time, several HCV proteins were shown to suppress apoptosis, thus preventing
the elimination of HCV-infected cells, and promoting virus persistence [2]. Dysregulation of
proapoptotic pathway,s such as the disruption of Bcl-2 and Bcl-xL proteins, facilitates cell proliferation,
possibly contributing to hepatocarcinogenesis [8]. Various HCV-encoded proteins play different roles
in modulating apoptosis by interacting and interfering with host factors, but literature data on the role
of individual HCV proteins in modulation of apoptosis are controversial [2,8–11]. HCV also promotes autophagy, as revealed both in vitro in various hepatoma cell lines and
in vivo by analyzing biopsies from CHC patients [1–3,12]. 1. Introduction Autophagy represents a conservative
regulated mechanism during which cell organelles are targeted for degradation in autolysosomes
in order to preserve cell homeostasis. Autophagy is generally regarded as a pro-survival event. However, intensive formation of autophagosomes, enhanced permeabilization of mitochondrial
membrane and critical decreases in ATP levels can trigger an “autophagy-associated cell death” [4]
similar to that observed during acute HCV infection [1,13]. Interplay between autophagy and
HCV infection remains controversial. On one hand, Sir et al. revealed a co-localization of the
main autophagy marker LC3 (microtubule associated protein 1 light chain 3) with HCV replicase
proteins on autophagosome membranes [14], and it was shown that inhibitors of autophagy suppress
HCV replication in cell culture [15,16]. One the other hand, Mohl et al. demonstrated that at least
NS5A, one of HCV replicase components, does not co-localize with autophagosomes during active
replication of the virus [17]. Several authors have proposed that HCV can initiate the formation of
autophagosomes, whereas this process can be blocked at later stages [18]. In support, HCV-induced
autophagy has been assumed to alter innate immune response and facilitate cell proliferation by
blocking apoptosis [19,20]. At the same time, others observed that HCV infection induced autophagy
accompanied by lysosomal activity [2]. It is assumed that a high viral load during HCV infection promotes hepatocyte death through
necrosis (or necroptosis, its regulated form), whereas low viral load can induce apoptosis [21,22]. Necrosis represents another type of cell death [4]. It is mediated by acute tissue injury, and is
characterized by the loss of cell membrane integrity with the concomitant release of metabolites
into the extracellular space. Necrosis triggers both a toxic response in the surrounding live cells and an
immune response. However, there are still only scarce data about the mechanisms of how the virus
triggers necrosis, and on the virus proteins that mediate this response. All the above-mentioned information shows that exact pathways which convey HCV-induced
metabolic and stress responses still remain questionable. There are debates about virus regulation of
the hepatocyte proliferation/death, and the role of individual proteins of the virus remains obscure. 2.1. Several HCV Proteins Hamper Huh7.5 Cell Growth First, an influence of HCV proteins on growth of Huh7.5 cells was analyzed. The cells were
transfected with the plasmids encoding individual proteins or with the NS3-NS5B polyprotein, were
stained with 4′,6-diamidino-2-phenylindole (DAPI) 72 h posttransfection, and their quantities were
estimated using a fluorescent microscope. Transfection efficacy was estimated by immunofluorescent
staining of the HCV protein in cells with monoclonal antibodies. The images are presented in
the Figure S1. In all cases, the efficacy exceeded 60% with no statistical differences between the
plasmids. Moreover, no pronounced difference was found in the intensity of fluorescence between
the cells’ expression of the individual HCV proteins, NS3-NS5B polyprotein, or cells harboring the
replicon. Analysis of the expression of NS3, NS5A, and NS5B proteins in cells expressing these proteins
and the ones with the replicon was performed by western blot analysis. It revealed that in the case of
the HCV replicon, the levels of NS5B were lower than in case of their overexpression (Figure S2). Int. J. Mol. Sci. 2017, 18, 2346
3 of 16
estimated using a fluorescent microscope. Transfection efficacy was estimated by immunofluorescent
staining of the HCV protein in cells with monoclonal antibodies. The images are presented in the
Figure S1. In all cases, the efficacy exceeded 60% with no statistical differences between the plasmids. Moreover, no pronounced difference was found in the intensity of fluorescence between the cells’
expression of the individual HCV proteins, NS3-NS5B polyprotein, or cells harboring the replicon. Analysis of the expression of NS3, NS5A, and NS5B proteins in cells expressing these proteins and The expression of NS4A/B and NS5A, as well as of NS3-NS5B polyprotein did not affect cell
growth and viability, compared to Huh7.5 cells transfected with the empty pcDNA3.1(+) vector
(Figure 1). In its turn, transfection with this vector did not alter the viability of cells, as compared to
the untransfected Huh7.5 cells. NS5B caused a minor (<20%) decrease in the number of living cells,
whereas E1/E2 glycoproteins and NS3 protein caused a more pronounced decrease in the number
of living cells (~30%). The highest decrease in the number of cells (47–56%) was observed in case of
the transient expression of the HCV core, and stable replication of the full-length genome of the virus
(the HCV replicon). Analysis of the expression of NS3, NS5A, and NS5B proteins in cells expressing these proteins and
the ones with the replicon was performed by western blot analysis. 1. Introduction The goal of this study was to estimate the impact of HCV proteins on autophagy, apoptosis,
and necrosis in the human hepatoma Huh7.5 cell line by analyzing the respective markers in cells
transiently expressing the core, E1/E2, NS3, NS4A/B, NS5A, and NS5B proteins individually or
simultaneously, as well as in cells harboring the HCV full-length replicon, and thus, expressing the
whole proteome at a higher physiological level. Int. J. Mol. Sci. 2017, 18, 2346 3 of 16 2. Results 2.1. Several HCV Proteins Hamper Huh7.5 Cell Growth Several hepatitis C virus (HCV) proteins hamper growth/viability of Huh7.5 cells. The cells,
transfected with plasmids encoding HCV proteins, were counted 72 h posttransfection by microscopy
visualization, and the values were normalized to the number of cells transfected with the empty
pcDNA3.1(+) vector. The data are presented as means ± standard error of the mean (SEM) of
eight measurements done in three independent experiments. * p < 0.05 vs. cells transfected with
pcDNA3.1(+) vector (black bar). 2.1. Several HCV Proteins Hamper Huh7.5 Cell Growth It revealed that in the case of the
HCV replicon, the levels of NS5B were lower than in case of their overexpression (Figure S2). The expression of NS4A/B and NS5A, as well as of NS3-NS5B polyprotein did not affect cell
growth and viability, compared to Huh7.5 cells transfected with the empty pcDNA3.1(+) vector
(Figure 1). In its turn, transfection with this vector did not alter the viability of cells, as compared to
the untransfected Huh7.5 cells. NS5B caused a minor (<20%) decrease in the number of living cells,
whereas E1/E2 glycoproteins and NS3 protein caused a more pronounced decrease in the number of
living cells (~30%). The highest decrease in the number of cells (47–56%) was observed in case of the
transient expression of the HCV core, and stable replication of the full-length genome of the virus
(the HCV replicon). Figure 1. Several hepatitis C virus (HCV) proteins hamper growth/viability of Huh7.5 cells. The cells,
transfected with plasmids encoding HCV proteins, were counted 72 h posttransfection by microscopy
visualization, and the values were normalized to the number of cells transfected with the empty
pcDNA3.1(+) vector. The data are presented as means ± standard error of the mean (SEM) of eight
measurements done in three independent experiments. * p < 0.05 vs. cells transfected with
pcDNA3.1(+) vector (black bar). 2.2. HCV Proteins Exhibit Different Regulatory Activity towards Apoptotic Pathways
Figure 1. Several hepatitis C virus (HCV) proteins hamper growth/viability of Huh7.5 cells. The cells,
transfected with plasmids encoding HCV proteins, were counted 72 h posttransfection by microscopy
visualization, and the values were normalized to the number of cells transfected with the empty
pcDNA3.1(+) vector. The data are presented as means ± standard error of the mean (SEM) of
eight measurements done in three independent experiments. * p < 0.05 vs. cells transfected with
pcDNA3.1(+) vector (black bar). Figure 1. Several hepatitis C virus (HCV) proteins hamper growth/viability of Huh7.5 cells. The cells,
transfected with plasmids encoding HCV proteins, were counted 72 h posttransfection by microscopy
visualization, and the values were normalized to the number of cells transfected with the empty
pcDNA3.1(+) vector. The data are presented as means ± standard error of the mean (SEM) of eight
measurements done in three independent experiments. * p < 0.05 vs. cells transfected with
pcDNA3.1(+) vector (black bar). 2.2. HCV Proteins Exhibit Different Regulatory Activity towards Apoptotic Pathways
Figure 1. 2. Results 2.1. Several HCV Proteins Hamper Huh7.5 Cell Growth Our next step was to investigate possible mechanisms of the apoptosis induction
expression of HCV proteins. The induction of apoptosis was accessed by quantifyin
2.2. HCV Proteins Exhibit Different Regulatory Activity towards Apoptotic Pathways NS5A protein reduced the number of the cells with the activated caspase-3, whereas
core, E1/E2, and NS3 proteins increased the rate of detection of the activated caspase by 1.6–2.6-fold. A similar increase (3.2-fold) was also observed in cells harboring the full-length HCV replicon. Int. J. Mol. Sci. 2017, 18, 2346
4 of 16
Activation of caspase-3 was detected in 3.9% Huh7.5 cells transfected with the empty vector
(Figure 2c). NS5A protein reduced the number of the cells with the activated caspase-3, whereas core, Activated caspase-8 was detected in 3.3% cells transfected with the empty vector (Figure 3d). Expressions of NS4A/B and NS5B proteins led to a decrease in the number of caspase-8 positive cells
by two-fold, whereas the HCV core, NS3, NS3-NS5B polyprotein and the virus replicon increased the
number of such cells by 3.1, 2.7, 1.8, and 1.8-fold, respectively, compared to the vector. (
g
)
p
p
E1/E2, and NS3 proteins increased the rate of detection of the activated caspase by 1.6–2.6-fold. A similar increase (3.2-fold) was also observed in cells harboring the full-length HCV replicon. Activated caspase-8 was detected in 3.3% cells transfected with the empty vector (Figure 3d). Expressions of NS4A/B and NS5B proteins led to a decrease in the number of caspase-8 positive cells
by two-fold, whereas the HCV core, NS3, NS3-NS5B polyprotein and the virus replicon increased the
u
be of u h ell by 3 1 2 7 1 8 a d 1 8 fold
e
e ti ely
o
a ed to the
e to number of such cells by 3.1, 2.7, 1.8, and 1.8 fold, respectively, compared to the vector. Figure 2. HCV proteins affect activation of caspases-3, -8 and -9 in Huh7.5 cells in different manners. (a) Immunofluorescent staining of the activated caspase-9 and HCV proteins in Huh7.5 cells
transiently expressing the HCV core or NS5A proteins, or harboring the full-length HCV replicon
(400 × magnification). Vertical panels left to right: staining with rabbit anti-caspase-9 primary and
anti-rabbit secondary antibodies conjugated to Cy3 (orange), merge with nuclear staining with DAPI
(blue), staining with mouse monoclonal antibodies to HCV proteins and anti-mouse secondary
antibodies conjugated to fluoresceine isothiocianate (FITC; green), combined with nuclear staining
with DAPI (blue). The arrows indicate caspase-9 positive cells. (b–d) Percentages of the cells which
tested positive for the caspases-9 (b), -3 (c), and -8 (d). Our next step was to investigate possible mechanisms of the apoptosis induction
expression of HCV proteins. The induction of apoptosis was accessed by quantifyin
2.2. HCV Proteins Exhibit Different Regulatory Activity towards Apoptotic Pathways caspases-3, -8, and -9 that mediate major apoptotic pathways. These activated caspases were detected
in the cytoplasm of the cells, using the specific antibodies, as homogenous intensive staining. Typical
images, exemplified in caspase-9, are presented in Figure 2a, and the quantification of the data for all
three caspases is given in the Figure 2b–d. Different caspases were present in the cells with different
rates of detection, depending on the HCV protein expressed. Caspase-9 was detected in 4.9% cells transfected with the empty vector control. Expression of
HCV NS5A and NS5B proteins reduced the number of the caspase-positive cells by two-fold, whereas
the core protein increased the number of cells with the activated caspase-9 by an additional 2.1-fold,
compared to the vector (Figure 2a,b). Expression of other HCV proteins, as well as of NS3-NS5B
Our next step was to investigate possible mechanisms of the apoptosis induction during the
expression of HCV proteins. The induction of apoptosis was accessed by quantifying activated
caspases-3, -8, and -9 that mediate major apoptotic pathways. These activated caspases were
detected in the cytoplasm of the cells, using the specific antibodies, as homogenous intensive staining. Typical images, exemplified in caspase-9, are presented in Figure 2a, and the quantification of the data
for all three caspases is given in the Figure 2b–d. Different caspases were present in the cells with
different rates of detection, depending on the HCV protein expressed. polyprotein, had no statistically significant effect. Finally, Huh7 cells harboring the HCV replicon
exhibited a 1.6-fold increase in the number of cells with the activated caspase, compared to the
control cells. Caspase-9 was detected in 4.9% cells transfected with the empty vector control. Expression of HCV
NS5A and NS5B proteins reduced the number of the caspase-positive cells by two-fold, whereas the Int. J. Mol. Sci. 2017, 18, 2346 4 of 16 core protein increased the number of cells with the activated caspase-9 by an additional 2.1-fold,
compared to the vector (Figure 2a,b). Expression of other HCV proteins, as well as of NS3-NS5B
polyprotein, had no statistically significant effect. Finally, Huh7 cells harboring the HCV replicon
exhibited a 1.6-fold increase in the number of cells with the activated caspase, compared to the
control cells. Activation of caspase-3 was detected in 3.9% Huh7.5 cells transfected with the empty vector
(Figure 2c). Our next step was to investigate possible mechanisms of the apoptosis induction
expression of HCV proteins. The induction of apoptosis was accessed by quantifyin
2.2. HCV Proteins Exhibit Different Regulatory Activity towards Apoptotic Pathways Values on each diagram are means ± SEM of
eight measurements done in three independent experiments, * p < 0.05 compared to the cells
transfected with the empty vector (black bar). Figure 2. HCV proteins affect activation of caspases-3, -8 and -9 in Huh7.5 cells in different
manners. (a) Immunofluorescent staining of the activated caspase-9 and HCV proteins in Huh7.5 cells
transiently expressing the HCV core or NS5A proteins, or harboring the full-length HCV replicon
(400× magnification). Vertical panels left to right: staining with rabbit anti-caspase-9 primary and
anti-rabbit secondary antibodies conjugated to Cy3 (orange), merge with nuclear staining with DAPI
(blue), staining with mouse monoclonal antibodies to HCV proteins and anti-mouse secondary
antibodies conjugated to fluoresceine isothiocianate (FITC; green), combined with nuclear staining
with DAPI (blue). The arrows indicate caspase-9 positive cells. (b–d) Percentages of the cells which
tested positive for the caspases-9 (b), -3 (c), and -8 (d). Values on each diagram are means ± SEM of
eight measurements done in three independent experiments, * p < 0.05 compared to the cells transfected
with the empty vector (black bar). Figure 2. HCV proteins affect activation of caspases-3, -8 and -9 in Huh7.5 cells in different manners. (a) Immunofluorescent staining of the activated caspase-9 and HCV proteins in Huh7.5 cells
transiently expressing the HCV core or NS5A proteins, or harboring the full-length HCV replicon
(400 × magnification). Vertical panels left to right: staining with rabbit anti-caspase-9 primary and
anti-rabbit secondary antibodies conjugated to Cy3 (orange), merge with nuclear staining with DAPI
(blue), staining with mouse monoclonal antibodies to HCV proteins and anti-mouse secondary
antibodies conjugated to fluoresceine isothiocianate (FITC; green), combined with nuclear staining
with DAPI (blue). The arrows indicate caspase-9 positive cells. (b–d) Percentages of the cells which
tested positive for the caspases-9 (b), -3 (c), and -8 (d). Values on each diagram are means ± SEM of
eight measurements done in three independent experiments, * p < 0.05 compared to the cells
transfected with the empty vector (black bar). Figure 2. HCV proteins affect activation of caspases-3, -8 and -9 in Huh7.5 cells in different
manners. (a) Immunofluorescent staining of the activated caspase-9 and HCV proteins in Huh7.5 cells
transiently expressing the HCV core or NS5A proteins, or harboring the full-length HCV replicon
(400× magnification). Our next step was to investigate possible mechanisms of the apoptosis induction
expression of HCV proteins. The induction of apoptosis was accessed by quantifyin
2.2. HCV Proteins Exhibit Different Regulatory Activity towards Apoptotic Pathways Vertical panels left to right: staining with rabbit anti-caspase-9 primary and
anti-rabbit secondary antibodies conjugated to Cy3 (orange), merge with nuclear staining with DAPI
(blue), staining with mouse monoclonal antibodies to HCV proteins and anti-mouse secondary
antibodies conjugated to fluoresceine isothiocianate (FITC; green), combined with nuclear staining
with DAPI (blue). The arrows indicate caspase-9 positive cells. (b–d) Percentages of the cells which
tested positive for the caspases-9 (b), -3 (c), and -8 (d). Values on each diagram are means ± SEM of
eight measurements done in three independent experiments, * p < 0.05 compared to the cells transfected
with the empty vector (black bar). 5 of 16
5 of 16 Int. J. Mol. Sci. 2017, 18, 2346
I
J M l S i
18 2346 DNA fragmentation, the end stage of apoptosis, was studied by the terminal deoxynucleotidyl
transferase dUTP nick end labeling (TUNEL) approach. Such staining of cells allowed the visualization
of a bright fluorescent signal in the nuclei of some cells (Figure 3a). In the Huh7.5 cells transfected
with the empty vector, 3.8% of the population exhibited signs of DNA fragmentation (Figure 3b). The increase in the number of TUNEL-positive cells was observed for the cells expressing the core
and E1/E2 proteins, or harboring the HCV replicon. In the case of other HCV proteins, no statistically
significant changes were found. DNA fragmentation, the end stage of apoptosis, was studied by the terminal deoxynucleotidyl
transferase dUTP nick end labeling (TUNEL) approach. Such staining of cells allowed the
visualization of a bright fluorescent signal in the nuclei of some cells (Figure 3a). In the Huh7.5 cells
transfected with the empty vector, 3.8% of the population exhibited signs of DNA fragmentation
(Figure 3b). The increase in the number of TUNEL-positive cells was observed for the cells expressing
the core and Е1/Е2 proteins, or harboring the HCV replicon. In the case of other HCV proteins, no
statistically significant changes were found. Figure 3. The HCV core and E1/E2 increase the number of Huh7.5 cells with nuclear DNA
fragmentation, i.e., at the end stage of apoptosis. (a) Huh7.5 cells transfected with the core- and E1/E2-
expressing plasmid or the empty pcDNA3.1 vector were stained 72 h posttransfection with the
“DeadEnd™ Fluorometric terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)
System” kit (green), with mouse monoclonal antibodies on HCV proteins and anti-mouse secondary
antibodies conjugated to Alexa Fluor 594 (AF594), and with DAPI. Our next step was to investigate possible mechanisms of the apoptosis induction
expression of HCV proteins. The induction of apoptosis was accessed by quantifyin
2.2. HCV Proteins Exhibit Different Regulatory Activity towards Apoptotic Pathways Vertical panels left to
right: TUNEL staining (green), HCV proteins (red), and overlay of TUNEL, HCV proteins and DAPI
staining; (b) Percentages of TUNEL-positive cells. Values are means ± SEM of eight measurements
done in three independent experiments, * p < 0.05 compared to the cells transfected with the empty
vector (black bar). Figure 3. The HCV core and E1/E2 increase the number of Huh7.5 cells with nuclear DNA
fragmentation, i.e., at the end stage of apoptosis. (a) Huh7.5 cells transfected with the core- and E1/E2-
expressing plasmid or the empty pcDNA3.1 vector were stained 72 h posttransfection with the
“DeadEnd™ Fluorometric terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)
System” kit (green), with mouse monoclonal antibodies on HCV proteins and anti-mouse secondary
antibodies conjugated to Alexa Fluor 594 (AF594), and with DAPI. Vertical panels left to right: TUNEL
staining (green), HCV proteins (red), and overlay of TUNEL, HCV proteins and DAPI staining;
(b) Percentages of TUNEL-positive cells. Values are means ± SEM of eight measurements done in
three independent experiments, * p < 0.05 compared to the cells transfected with the empty vector
(black bar). Figure 3. The HCV core and E1/E2 increase the number of Huh7.5 cells with nuclear DNA
fragmentation, i.e., at the end stage of apoptosis. (a) Huh7.5 cells transfected with the
core- and E1/E2-expressing plasmid or the empty pcDNA3.1 vector were stained 72 h posttransfection
with the “DeadEnd™Fluorometric terminal deoxynucleotidyl transferase dUTP nick end labeling
(TUNEL) System” kit (green), with mouse monoclonal antibodies on HCV proteins and anti-mouse
secondary antibodies conjugated to Alexa Fluor 594 (AF594), and with DAPI. Vertical panels left to
right: TUNEL staining (green), HCV proteins (red), and overlay of TUNEL, HCV proteins and DAPI
staining; (b) Percentages of TUNEL-positive cells. Values are means ± SEM of eight measurements
done in three independent experiments, * p < 0.05 compared to the cells transfected with the empty
vector (black bar). Our next step was to investigate possible mechanisms of the apoptosis induction
expression of HCV proteins. The induction of apoptosis was accessed by quantifyin
2.2. HCV Proteins Exhibit Different Regulatory Activity towards Apoptotic Pathways Vertical panels left to right: TUNEL
staining (green), HCV proteins (red), and overlay of TUNEL, HCV proteins and DAPI staining;
(b) Percentages of TUNEL-positive cells. Values are means ± SEM of eight measurements done in
three independent experiments, * p < 0.05 compared to the cells transfected with the empty vector
(black bar). Figure 3. The HCV core and E1/E2 increase the number of Huh7.5 cells with nuclear DNA
fragmentation, i.e., at the end stage of apoptosis. (a) Huh7.5 cells transfected with the
core- and E1/E2-expressing plasmid or the empty pcDNA3.1 vector were stained 72 h posttransfection
with the “DeadEnd™Fluorometric terminal deoxynucleotidyl transferase dUTP nick end labeling
(TUNEL) System” kit (green), with mouse monoclonal antibodies on HCV proteins and anti-mouse
secondary antibodies conjugated to Alexa Fluor 594 (AF594), and with DAPI. Vertical panels left to
right: TUNEL staining (green), HCV proteins (red), and overlay of TUNEL, HCV proteins and DAPI
staining; (b) Percentages of TUNEL-positive cells. Values are means ± SEM of eight measurements
done in three independent experiments, * p < 0.05 compared to the cells transfected with the empty
vector (black bar). Figure 3. The HCV core and E1/E2 increase the number of Huh7.5 cells with nuclear DNA
fragmentation, i.e., at the end stage of apoptosis. (a) Huh7.5 cells transfected with the core- and E1/E2-
expressing plasmid or the empty pcDNA3.1 vector were stained 72 h posttransfection with the
“DeadEnd™ Fluorometric terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)
System” kit (green), with mouse monoclonal antibodies on HCV proteins and anti-mouse secondary
antibodies conjugated to Alexa Fluor 594 (AF594), and with DAPI. Vertical panels left to right: TUNEL
staining (green), HCV proteins (red), and overlay of TUNEL, HCV proteins and DAPI staining;
(b) Percentages of TUNEL-positive cells. Values are means ± SEM of eight measurements done in
three independent experiments, * p < 0.05 compared to the cells transfected with the empty vector
(black bar). Figure 3. The HCV core and E1/E2 increase the number of Huh7.5 cells with nuclear DNA
fragmentation, i.e., at the end stage of apoptosis. (a) Huh7.5 cells transfected with the
core- and E1/E2-expressing plasmid or the empty pcDNA3.1 vector were stained 72 h posttransfection
with the “DeadEnd™Fluorometric terminal deoxynucleotidyl transferase dUTP nick end labeling
(TUNEL) System” kit (green), with mouse monoclonal antibodies on HCV proteins and anti-mouse
secondary antibodies conjugated to Alexa Fluor 594 (AF594), and with DAPI. 2.3. Several HCV Proteins Stimulate Autophagy in Huh7.5 Cells
2.3. Several HCV Proteins Stimulate Autophagy in Huh7.5 Cells 2017, 18, 2346 6 of 16 Additional verification of these results was performed by the staining of microtubule-associated
protein 1A/1B-light chain 3 (LC3). During the formation of autophagosomes, LC3 protein is conjugated
to phosphatidylethanolamine, leading to the accumulation of this product and an increase in the level
of fluorescence. In our experiments, we observed an increase in the number of cells with enhanced
fluorescence in punctuate structures in the cytoplasm in the case of the expression of HCV proteins
(as is exemplified in Figure 4c). Among Huh7.5 cells transfected with the empty vector, only 3.7%
tested positive for LC3. The expression of E1/E2, NS4A/B, NS5A, and NS5B proteins led to the
1.3–1.6-fold increase in LC3–positive cells (Figure 4d). In the case of cells harboring the HCV replicon,
>8% of cells demonstrated bright fluorescence. The HCV core had no effect on the number of cells
with strong LC3 staining, whereas the expression of NS3 or NS3-NS5B resulted in a tendency of the
polyprotein to increase (p = 0.15 and 0.09, respectively). Thus, a comparison of the LC3 staining and
MDC accumulation revealed similar results, with LC3 staining being less sensitive. Int. J. Mol. Sci. 2017, 18, 2346
6 of 16
enhanced fluorescence in punctuate structures in the cytoplasm in the case of the expression of HCV
proteins (as is exemplified in Figure 4c). Among Huh7.5 cells transfected with the empty vector, only
3.7% tested positive for LC3. The expression of E1/E2, NS4A/B, NS5A, and NS5B proteins led to the
1.3–1.6-fold increase in LC3–positive cells (Figure 4d). In the case of cells harboring the HCV replicon,
>8% of cells demonstrated bright fluorescence. The HCV core had no effect on the number of cells
with strong LC3 staining, whereas the expression of NS3 or NS3-NS5B resulted in a tendency of the
polyprotein to increase (p = 0.15 and 0.09, respectively). Thus, a comparison of the LC3 staining and
MDC a
u
ulatio
e ealed i
ila
e ult
ith LC3 tai i
bei
le
e
iti e Figure 4. HCV proteins activated autophagy in Huh7.5 cells, as revealed by the enhanced
incorporation of monodansylcadaverine into autophagosomes and detection of LC3. (a) Huh7.5 cells
harboring the HCV replicon, or transfected with the E1/E2-expressing plasmid or the empty
pcDNA3.1(+) vector were stained 72 h posttransfection with the monodansylcadaverine (MDC) and
with DAPI. 2.3. Several HCV Proteins Stimulate Autophagy in Huh7.5 Cells
2.3. Several HCV Proteins Stimulate Autophagy in Huh7.5 Cells Autophagy was assessed by visualizing the incorporation of monodansylcadaverine (MDC) into
the living cells using an appropriate kit from Abcam. The cells with activated autophagy were
identified as cells with a bright fluorescent signal in the cytoplasm (Figure 4a). Among the Huh7.5
cells transfected with the pcDNA3.1(+) vector, 3.1% of the cells exhibited signs of autophagy
(Figure 4b). The expression of most of the studied HCV proteins (E1/E2, NS3, NS4A/B, NS5A, and
NS5B, as well as the NS3-NS5B polyprotein) independently or in the context of the full-length
replicon led to a 1.8–9.3-fold increase of MDC incorporation. The only virus protein that did not affect
the incidence of autophagy was the HCV core. To verify this result, the cells transfected either with
the vector or with the core-expressing plasmid were incubated with 60 µM chloroquine, an
autophagy inhibitor, or with 250 nM rapamycin, a standard autophagy inducer, for 3 h. Indeed,
chloroquine suppressed MDC incorporation in both cases by 2.1–2.7-fold. In contrast, rapamycin
augmented the autophagy incidence by 1.8–2.3-fold. Thus, the HCV core neither provoked
autophagy nor inhibited it, if induced by other stimuli. Additi
l
ifi
ti
f th
lt
f
d b th
t i i
f
i
t b l
i t d
Autophagy was assessed by visualizing the incorporation of monodansylcadaverine (MDC)
into the living cells using an appropriate kit from Abcam. The cells with activated autophagy were
identified as cells with a bright fluorescent signal in the cytoplasm (Figure 4a). Among the Huh7.5 cells
transfected with the pcDNA3.1(+) vector, 3.1% of the cells exhibited signs of autophagy (Figure 4b). The expression of most of the studied HCV proteins (E1/E2, NS3, NS4A/B, NS5A, and NS5B, as well
as the NS3-NS5B polyprotein) independently or in the context of the full-length replicon led to a
1.8–9.3-fold increase of MDC incorporation. The only virus protein that did not affect the incidence
of autophagy was the HCV core. To verify this result, the cells transfected either with the vector or
with the core-expressing plasmid were incubated with 60 µM chloroquine, an autophagy inhibitor,
or with 250 nM rapamycin, a standard autophagy inducer, for 3 h. Indeed, chloroquine suppressed
MDC incorporation in both cases by 2.1–2.7-fold. In contrast, rapamycin augmented the autophagy
incidence by 1.8–2.3-fold. Thus, the HCV core neither provoked autophagy nor inhibited it, if induced
by other stimuli. Int. J. Mol. Sci. 2.3. Several HCV Proteins Stimulate Autophagy in Huh7.5 Cells
2.3. Several HCV Proteins Stimulate Autophagy in Huh7.5 Cells Vertical panels from the left to the right are: MDC staining (green), nuclear staining with
DAPI (blue), overlay of MDC and DAPI staining; (b) Percentages of MDC-positive cells. (c) immunofluorescent staining of the activated LC3 and NS5B protein in Huh7.5 cells (400×
magnification). Vertical panels left to right: staining with rabbit anti-LC3 primary and anti-rabbit
secondary antibodies conjugated to Cy3 (orange) or primary mouse anti-NS5B antibody and
secondary anti-mouse antibodies conjugated to FITC (green), merge with nuclear staining with DAPI
(blue) The arrows indicate cells with LC3 punctuate staining; (d) Percentages of LC3-positive cells. Values are means ± SEM of eight measurements done in three independent experiments, * p < 0.05
compared to the cells transfected with the empty vector (black bar). Figure 4. HCV proteins activated autophagy in Huh7.5 cells, as revealed by the enhanced incorporation
of monodansylcadaverine into autophagosomes and detection of LC3. (a) Huh7.5 cells harboring the
HCV replicon, or transfected with the E1/E2-expressing plasmid or the empty pcDNA3.1(+) vector were
stained 72 h posttransfection with the monodansylcadaverine (MDC) and with DAPI. Vertical panels
from the left to the right are: MDC staining (green), nuclear staining with DAPI (blue), overlay of
MDC and DAPI staining; (b) Percentages of MDC-positive cells. (c) immunofluorescent staining of
the activated LC3 and NS5B protein in Huh7.5 cells (400× magnification). Vertical panels left to right:
staining with rabbit anti-LC3 primary and anti-rabbit secondary antibodies conjugated to Cy3 (orange)
or primary mouse anti-NS5B antibody and secondary anti-mouse antibodies conjugated to FITC (green),
merge with nuclear staining with DAPI (blue) The arrows indicate cells with LC3 punctuate staining;
(d) Percentages of LC3-positive cells. Values are means ± SEM of eight measurements done in three
independent experiments, * p < 0.05 compared to the cells transfected with the empty vector (black bar). Figure 4. HCV proteins activated autophagy in Huh7.5 cells, as revealed by the enhanced
incorporation of monodansylcadaverine into autophagosomes and detection of LC3. (a) Huh7.5 cells
harboring the HCV replicon, or transfected with the E1/E2-expressing plasmid or the empty
pcDNA3.1(+) vector were stained 72 h posttransfection with the monodansylcadaverine (MDC) and
with DAPI. Vertical panels from the left to the right are: MDC staining (green), nuclear staining with
DAPI (blue), overlay of MDC and DAPI staining; (b) Percentages of MDC-positive cells. (c) immunofluorescent staining of the activated LC3 and NS5B protein in Huh7.5 cells (400×
magnification). 3. Discussion
3. Discussion In the current study we present an analysis of the influence of expression of various structural
and non-structural HCV proteins on the viability/proliferation of hepatoma Huh7.5 cells. It is a well-
established fact that chronic hepatitis C is accompanied by the continuous death of infected
hepatocytes. It has been shown in the context of both the liver of chronic hepatitis C patients and in
vitro in the infected primary or immortalized hepatocytes [10,21]. However, the input of individual
proteins of the virus in the modulation of cell fate still remains disputable. In order to investigate
mechanisms of cell death in response to the expression of HCV proteins, we used an array of
immunocytochemical approaches for the detection of five markers of apoptosis and autophagy, as
well as a cytological approach for necrosis detection. The results are summarized in Table 1. In the current study we present an analysis of the influence of expression of various structural
and non-structural HCV proteins on the viability/proliferation of hepatoma Huh7.5 cells. It is a
well-established fact that chronic hepatitis C is accompanied by the continuous death of infected
hepatocytes. It has been shown in the context of both the liver of chronic hepatitis C patients
and in vitro in the infected primary or immortalized hepatocytes [10,21]. However, the input of
individual proteins of the virus in the modulation of cell fate still remains disputable. In order to
investigate mechanisms of cell death in response to the expression of HCV proteins, we used an array
of immunocytochemical approaches for the detection of five markers of apoptosis and autophagy,
as well as a cytological approach for necrosis detection. The results are summarized in Table 1. Table 1. Impact of HCV proteins on promoting various types of death of Huh7.5 hepatoma cells. Table 1. Impact of HCV proteins on promoting various types of death of Huh7.5 hepatoma cells. Table 1. Impact of HCV proteins on promoting various types of death of Huh7.5 hepatoma cells. 2.4. HCV NS5A Protein Moderately Increases the Incidence Rate of Necrosis
2.4. HCV NS5A Protein Moderately Increases the Incidence Rate of Necrosis Values are means ± SEM from three independent experiments, * p < 0.05
compared to the cells transfected with the empty vector (black bar). 2.4. HCV NS5A Protein Moderately Increases the Incidence Rate of Necrosis
2.4. HCV NS5A Protein Moderately Increases the Incidence Rate of Necrosis Necrotic cell death was evaluated by determining damage to the plasma membrane. Among the
control Huh7.5 cells transfected with the pcDNA3.1(+) vector, not more that 1.6% demonstrated signs
of necrosis. A statistically significant 2.1-fold increase in the number of necrotic cells was observed only
in the case of the expression of the HCV NS5A protein: in this case 3.3% necrotic cells were detected
(Figure 5). Necrotic cell death was evaluated by determining damage to the plasma membrane. Among the
control Huh7.5 cells transfected with the pcDNA3.1(+) vector, not more that 1.6% demonstrated signs
of necrosis. A statistically significant 2.1-fold increase in the number of necrotic cells was observed
only in the case of the expression of the HCV NS5A protein: in this case 3.3% necrotic cells were
detected (Figure 5). Figure 5. HCV NS5A protein promotes necrotic cell death. Huh7.5 cells harboring the HCV replicon,
or transfected with the HCV-expressing plasmid or the empty pcDNA3.1(+) vector were stained 72 h
posttransfection with trypan blue, and the stained cells were counted. The bars represent percentages
of trypan-positive cells. Values are means ± SEM from three independent experiments, * p < 0.05
compared to the cells transfected with the empty vector (black bar). Figure 5. HCV NS5A protein promotes necrotic cell death. Huh7.5 cells harboring the HCV replicon,
or transfected with the HCV-expressing plasmid or the empty pcDNA3.1(+) vector were stained 72 h
posttransfection with trypan blue, and the stained cells were counted. The bars represent percentages
of trypan-positive cells. Values are means ± SEM from three independent experiments, * p < 0.05
compared to the cells transfected with the empty vector (black bar). Figure 5. HCV NS5A protein promotes necrotic cell death. Huh7.5 cells harboring the HCV replicon,
or transfected with the HCV-expressing plasmid or the empty pcDNA3.1(+) vector were stained 72 h
posttransfection with trypan blue, and the stained cells were counted. The bars represent percentages
of trypan-positive cells. Values are means ± SEM from three independent experiments, * p < 0.05
compared to the cells transfected with the empty vector (black bar). Figure 5. HCV NS5A protein promotes necrotic cell death. Huh7.5 cells harboring the HCV replicon,
or transfected with the HCV-expressing plasmid or the empty pcDNA3.1(+) vector were stained 72 h
posttransfection with trypan blue, and the stained cells were counted. The bars represent percentages
of trypan-positive cells. Necrosis
∼
∼
∼
∼
2.1
∼
∼
∼
1 Changes in rates of detection of the respective marker (fold). 2 Symbol “∼” denotes absence of a
1 Changes in rates of detection of the respective marker (fold). 2 Symbol “~” denotes absence of a statistically
significant effect. 2.3. Several HCV Proteins Stimulate Autophagy in Huh7.5 Cells
2.3. Several HCV Proteins Stimulate Autophagy in Huh7.5 Cells Vertical panels left to right: staining with rabbit anti-LC3 primary and anti-rabbit
secondary antibodies conjugated to Cy3 (orange) or primary mouse anti-NS5B antibody and
secondary anti-mouse antibodies conjugated to FITC (green), merge with nuclear staining with DAPI
(blue) The arrows indicate cells with LC3 punctuate staining; (d) Percentages of LC3-positive cells. Values are means ± SEM of eight measurements done in three independent experiments, * p < 0.05
compared to the cells transfected with the empty vector (black bar). Figure 4. HCV proteins activated autophagy in Huh7.5 cells, as revealed by the enhanced incorporation
of monodansylcadaverine into autophagosomes and detection of LC3. (a) Huh7.5 cells harboring the
HCV replicon, or transfected with the E1/E2-expressing plasmid or the empty pcDNA3.1(+) vector were
stained 72 h posttransfection with the monodansylcadaverine (MDC) and with DAPI. Vertical panels
from the left to the right are: MDC staining (green), nuclear staining with DAPI (blue), overlay of
MDC and DAPI staining; (b) Percentages of MDC-positive cells. (c) immunofluorescent staining of
the activated LC3 and NS5B protein in Huh7.5 cells (400× magnification). Vertical panels left to right:
staining with rabbit anti-LC3 primary and anti-rabbit secondary antibodies conjugated to Cy3 (orange)
or primary mouse anti-NS5B antibody and secondary anti-mouse antibodies conjugated to FITC (green),
merge with nuclear staining with DAPI (blue) The arrows indicate cells with LC3 punctuate staining;
(d) Percentages of LC3-positive cells. Values are means ± SEM of eight measurements done in three
independent experiments, * p < 0.05 compared to the cells transfected with the empty vector (black bar). Int. J. Mol. Sci. 2017, 18, 2346
Int J Mol Sci 2017 18 2346 7 of 16
7 of 16 2.4. HCV NS5A Protein Moderately Increases the Incidence Rate of Necrosis
2.4. HCV NS5A Protein Moderately Increases the Incidence Rate of Necrosis 3. Discussion
3. Discussion Markers of Cell Death
HCV Proteins
Core
E1/E2
NS3
NS4A/B
NS5A
NS5B
NS3-
NS5B
HCV
Replicon
Apoptosis
Caspase-8
3.1 1
∼ 2
2.7
0.5
∼
0.5
1.8
1.9
Caspase-9
2.1
∼
∼
∼
0.5
0.5
∼
1.6
Caspase-3
1.6
1.6
2.6
∼
0.6
∼
∼
3.2
TUNEL
2.0
2.2
∼
∼
∼
∼
∼
2.1
Auto-
phagy
MDC
∼
4.1
1.8
2.9
2.9
4.8
2.1
9.3
LC3-II
∼
1.3
∼
1.6
1.4
1.6
∼
2.2
Table 1. Impact of HCV proteins on promoting various types of death of Huh7.5 hepatoma cells. Markers of Cell Death
HCV Proteins
Core
E1/E2
NS3
NS4A/B
NS5A
NS5B
NS3-NS5B
HCV Replicon
Apoptosis
Caspase-8
3.1 1
~ 2
2.7
0.5
~
0.5
1.8
1.9
Caspase-9
2.1
~
~
~
0.5
0.5
~
1.6
Caspase-3
1.6
1.6
2.6
~
0.6
~
~
3.2
TUNEL
2.0
2.2
~
~
~
~
~
2.1
Auto-phagy
MDC
~
4.1
1.8
2.9
2.9
4.8
2.1
9.3
LC3-II
~
1.3
~
1.6
1.4
1.6
~
2.2
Necrosis
~
~
~
~
2.1
~
~
~
1
2 Int. J. Mol. Sci. 2017, 18, 2346 8 of 16 The results suggest that the overexpression of core and E1/E2 proteins as well as the continuous
replication of the virus genome RNA trigger apoptosis (Table 1). The core protein induced the activation
of initiator caspases-8 and -9, with subsequent activation of the effector caspase-3 that triggered DNA
fragmentation and cell death. The involvement of the HCV core in the receptor-mediated apoptosis is
in agreement with our previous findings that this protein triggers the secretion of tumor necrosis factor
α (TNF-α) when expressed in Huh7.5 cells [23]. Moreover, the data of the current study confirm several
reports from other groups about the pro-apoptotic effect of the core protein [8,24,25]. In addition to cell
systems, induction of apoptosis by the HCV core was also described in vivo in transgenic mice [26–28]. Core-mediated apoptosis can resulte from the activation of the mitochondrial signaling pathway
involving activation of the Bax (Bcl-2-like protein 4) protein via the disruption of calcium homeostasis
and concomitant mitochondria dysfunction, as well as from the TNF-dependent pathway [2,24,29]. However, the HCV core was also reported to block apoptosis by various mechanisms thus contributing
to core-driven hepatocarcinogenesis [8,30,31]. 3. Discussion
3. Discussion The discrepancy of these data could be explained
by a “competitive model” of the adaptive cell response to the stress, meaning that both lethal and
pro-survival programs took place with the overall effect depending on variability in experimental
settings in vitro [32]. It could be speculated that both events could take place in the liver: dying
hepatocytes could contribute to the development of inflammation and liver damage, whereas survival
of the stressed infected cells predisposes them to the malignant transformation. We observed that the surface E1/E2 glycoproteins increased the incidence of the activated
caspase-3 and concomitant apoptotic cell death (Table 1). At the same time, we failed to detect
any statistically significant enhancement in the activation of initiator caspases-8 and -9. Further studies
of the mechanisms of E1/E2-mediated cell death should be considered. The data on the involvement of
these proteins in apoptosis is scarce. However, most authors conclude that E1/E2 pro-apoptotic actions
during their expression in various cell systems takes place due to both outer TNF-dependent [33] and
inner mitochondrial pathways [33–35]. Since we previously observed the enhanced TNF-α production
in E1/E2-overexpressing Huh7.5 cells, apoptosis is likely to be mediated by an outer pathway [23]. Therefore, both glycoproteins may trigger apoptosis via other signaling cascades. It may be assumed
that apoptosis in such cells can be triggered by the activation of caspase-4, which does not affect the
status of caspase-3 [36], or via caspase-12, which activates both caspase-3 and -9 [37]. These caspases
can be activated by ER stress and unfolded protein response [36,37] that accompany the expression of
E1 and E2 glycoproteins [38]. However, this assumption merits further studies. The NS3 protein was shown to trigger the activation of caspase-3 via the activation of
caspase-8 through the outer apoptotic pathway (Table 1). This is in agreement with our previous
findings of the enhanced accumulation of TNF-α in the cells overexpressing NS3 protease [23]. However, this process does not lead to the accomplishment of apoptosis, since no DNA fragmentation
was observed. Activation of the caspase-8 in the presence of the NS3 protein was already reported [39]. Epithelial cells and fibroblasts, exposed to the recombinant NS3 protein, exhibited enhanced production
of pro-inflammatory cytokines, augmented reactive oxygen species (ROS) production thus leading
to apoptosis [25]. In contrast, NS3 was also shown to demonstrate anti-apoptotic activity that was
realized through interaction with tumor suppressor p53 [40]. 3. Discussion
3. Discussion Another aspect of our study was an investigation of the NS3-NS5B polyprotein effect on cell
death. Interestingly, a combination of the non-structural proteins activated caspase-8 with no effect on
DNA fragmentation, which takes place during apoptosis (Table 1). Since individual HCV proteins
exhibited different actions on apoptosis, it could be assumed that the pro-apoptotic activity of NS3 may
be balanced by the anti-apoptotic actions of NS4A/B, NS5A, and NS5B. In addition, the expression of
NS3 and NS5A proteins, along with other virus proteins, changes their localization, in contrast with
the overexpression of the individual proteins [46,47]. Finally, in similar conditions, Lim et al. reported
the activation of caspase-3 in response to the expression of NS3-NS5B polyprotein in primary human
hepatocytes (PHH) but not in hepatoma Huh7 cells, suggesting that Huh7.5, being tumor-derived
cells, are more resistant to pro-apoptotic stimuli [21]. Our study revealed that the expression of HCV proteins in the full-length replicon leads
to the activation of both the apoptotic pathways. Previously it was reported that Huh7.5 cells
harboring the subgenomic replicon or infected by recombinant variants of the virus in the HCV cell
culture (HCVcc) system activated FAS- and TRAIL-mediated outer and inner pathways of apoptosis
through the induction of the unfolded protein response, pro-apoptotic Bax protein and cytochrome
c release [2,46,48]. As the same was observed only for the core protein, this protein has the most
pronounced impact on apoptosis induction in HCV-infected cells. Autophagy in the living cells was analyzed by measuring the accumulation of fluorescent
monodansylcadaverine in autolysosomes that are formed during late stages of autophagy when
autophagosomes fuse with lysosomes. Moreover, MDC is co-localized with the main marker of
autophagosomes—the LC3 protein which can be detected during all stages of autophagosome
formation [49,50]. Enhanced autophagy in Huh7.5 cells in response to the expression of E1/E2,
NS4A/B, NS5A, NS5B proteins individually or in the context of the full-length replicon was revealed
by two independent approaches (Table 1). Differences were found in the case of NS3 protein and
NS3-NS5B polyprotein, for which only a tendency to statistically significant accumulation of LC3 was
observed. This could be due to a lower sensitivity of this approach, compared to the visualization of
MDC accumulation, and due to the fact that these autophagy markers are different in kind. 3. Discussion
3. Discussion Our data indicate that the individual non-structural proteins NS4A/B, NS5A and NS5B
suppressed various stages of activation of the caspase cascade (Table 1). NS5A prevented the activation
of the caspase-3, presumably through the inhibition of caspase-9. Notably, this is in agreement with
the literature data, as this protein was shown earlier to use multiple strategies to block inner pathways
of apoptosis by interfering with various host factors [8,9,41]. Since the inner pathway of apoptosis is
activated through mitochondrial dysfunction, the reason why NS5A did not induce it while the HCV
core did does not seem obvious; both of them triggered the massive production of ROS [42,43]. It can
be speculated that such difference is due to the different actions of these proteins on the ROS-producing
systems of the cell. We have recently demonstrated that enhanced productionof reactive oxygen species
(ROS) in NS5A-expressing cells does not result from the efflux of calcium ions from endoplasmic Int. J. Mol. Sci. 2017, 18, 2346 9 of 16 reticulum (ER) and their accumulation in mitochondria [44] that occur in the case of the HCV core [29]. However, other possible mechanisms accounting for these events cannot be excluded. reticulum (ER) and their accumulation in mitochondria [44] that occur in the case of the HCV core [29]. However, other possible mechanisms accounting for these events cannot be excluded. The NS5B protein exhibited a moderate suppressive effect towards the initiation of caspases-8
and -9 (Table 1), thus inhibiting apoptosis. There are only scarce data about the role of this protein in
the development of apoptosis. The pro-apoptotic activity of NS5B was assigned to its ability to interact
with the pro-apoptotic BIK protein [11], whereas its anti-apoptotic activity was assigned to its ability to
suppress NF-κB-dependent anti-apoptotic Bcl-xL and XIAP proteins during treatment with TNF-α [45]. Such a discrepancy could result from the usage of different cell models and expression vectors. Huh7.5 cells expressing NS4A/B showed the suppression of caspase-8 activation while other
apoptotic markers remained unaffected (Table 1). The literature reports that individual NS4A and
NS4B proteins can either stimulate or block apoptosis, depending on the cell model and the expression
status (transient or stable expression) [10,45,46]. In our experiments, NS4A was expressed together
with NS4B, and we failed to find any data on the effect of such combination on cell death. 3. Discussion
3. Discussion In general,
these data are in agreement with previous findings of the other groups that showed enhancement
of autophagy by the hepatitis C virus [1–3,31,51]. For example, Chu et al. also reported that the
induction of autophagy is achieved independently of HCV core expression, but that it is dependent
on nonstructural proteins [51]. The difference between our data and literature data is with regards
to the impact of individual proteins of the virus on this process. We show that the formation of
autophagosomes is promoted mostly by NS5B and E1/E2 proteins, whereas other groups reported
activation of autophagy by NS5A [18,52], NS4B [53], the core, NS3/4A and NS4B [54]. Though analysis of apoptosis and autophagy was performed by the quantification of biochemical
markers, necrotic cells with altered plasma membranes were visually detected. The increased number
of necrotic cells was detected only when the NS5A protein was expressed (Table 1). Notably, NS5A 10 of 16 Int. J. Mol. Sci. 2017, 18, 2346 10 of 16 exhibited no statistically significant effect on the incidence of necrosis when expressed with other
nonstructural proteins (i.e., in cells transfected with the plasmid encoding NS3-NS5B, or harboring
the HCV replicon). This could be due to several reasons including different expression levels of
NS5A, or its different localization. However, a comparison by Western blotting between accumulation
levels of NS5A in cells transfected with pcNS5A, and those harboring the HCV replicon revealed
no significant differences (Figure S2). Thus, NS5A may have a different distribution between
organelles when expressed individually or together with other HCV proteins. Nevertheless, our
results indicate that at least a part of the NS5A-expressing cells dies by necrosis. An impact
of necrosis on the death of the HCV-infected hepatocytes is still not fully understood. To our
knowledge, there are no data showing the promotion of necrosis by the hepatitis C virus and its
proteins. The only exception is the data of Lim et al., who demonstrated the occurrence of necroptosis
(i.e., a programmed necrosis) in the cells expressing HCV proteins, and its prevention by its inhibitor
necrostatin-1 [21]. Interestingly, in HCV-unrelated studies, caspase-8 was shown to inhibit necrosis [55]. Nevertheless, it is worth noting that in our study, necrosis was detected only in NS5A-expressing cells
where no activation of caspase-8 was observed (Table 1). 3. Discussion
3. Discussion We assume that different effects of HCV proteins on the induction of apoptosis, necrosis,
and autophagy do not result from different levels of their expression. For example, the activation of
caspases and DNA fragmentation was more pronounced for the core protein than for NS5A and NS5B
proteins. Previously we quantified the expression of these proteins in Huh7 cells and revealed that
HCV the core is accumulated up to 100 fg/cell, whereas NS5A and NS5B reached levels of 700 fg/cell
each [43]. The same levels of these nonstructural proteins also do not correlate with their different
effects on necrosis. Finally, the cells harboring the replicon exhibited higher rate of apoptotic cell death
than NS5B-expressing cells despite lower levels of its expression (Figure S2). The presented results show that HCV proteins expressed alone or in cells harboring the replicon
promote cell death by several pathways. At the same time, Pietschmann et al. established that the
HCV subgenomic replicon can propagate for up to 6 months without complete loss in Huh7 cells, even
in the absence of G418 [56]. Our data do not contradict that paper. First, a decrease in viral RNA levels
during cell passaging showed a moderate negative effect of the virus on the cells. Second, in our case
we used the full-length HCV replicon (which encoded the core protein) compared to the subgenomic
replicon of that study. It is accepted that the HCV core is more hazardous for Huh7 cells among all
the proteins of the virus. Third, in our hands, less than 10% of cells exhibited activation of caspases,
signs of pronounced DNA fragmentation, or of necrosis. Thus, in vitro it correlates with slower the
cell growth evidenced by many groups. In the context of infection, it also falls in line with an increased
incidence of hepatocyte death [57]. 4.1. Cells The naive Huh7.5 cell line [58] was cultured in Dulbecco’s modified minimal essential
medium
(DMEM)
(Paneco,
Moscow,
Russia)
supplemented
with
10%
fetal
calf
serum
(Gibco, Waltham, MA, USA), 2 mM glutamine, 50 µg/mL gentamycin at 37 ◦C in a humid atmosphere
with 5% CO2 · Huh7 cells harboring a bicistronic full-length I389/core-3′/5.1 HCV replicon [59]
were cultured similarly in DMEM/F12 medium (Paneco) containing fetal calf serum, glutamine and
supplemented with 300 µg/mL geneticin (G418, Invitrogen, Waltham, MA, USA). 4.5. Necrosis Necrosis was accessed using a widely used trypan blue assay that stains the cells with altered
membrane permeability [64,65]. Briefly, the cells were treated with trypan blue and the stained cells
were counted on a microscope using a Goryaev chamber. 4.2. Plasmids The plasmids encoding individual HCV cores (pcCore), E1 and E2 (pcE1E2), NS3 (pcNS3),
NS4A and NS4B (pcNS4), NS5A (pcNS5A) and NS5B (pcNS5B) were described previously [43,60–62]. In addition, a plasmid pcNS3-NS5B encoding NS3, NS4, NS5A and NS5B as a single polyprotein was
used [61]. All these plasmids were constructed using pcDNA3.1(+) vector (Invitrogen). The plasmids Int. J. Mol. Sci. 2017, 18, 2346 11 of 16 were propagated in XL-1 blue Escherichia coli strain and purified using QIAGEN® Plasmid Purification
Maxi Kit (Qiagen Inc., Hilden, Germany). were propagated in XL-1 blue Escherichia coli strain and purified using QIAGEN® Plasmid Purification
Maxi Kit (Qiagen Inc., Hilden, Germany). 4.3. Transfection Twenty four hours prior to transfection Huh7.5 cells were seeded in a 24-well plate
(SPL Lifesciences, Pocheon, Korea) with a microscope cover glass at a density of 5 × 104 cells/well. The transfections were carried out using TurboFect Transfection Reagent (Thermo Fisher Scientific,
Rockford, IL, USA) according to manufacturer’s specification. Seventy two hours posttransfection the
cells were washed with PBS and either studied directly or fixed as described below. 4.4. Immunocytochemical Analysis HCV proteins and markers of cell death in the transfected cells were detected by
immunocytochemical analysis. For detection of apoptosis and autophagy primary rabbit antibodies to
activated caspases-3, -8, -9, and LC3A/B (ab52293, ab4052, ab32539, and ab128025, respectively, Abcam,
Cambridge, UK), and secondary Cyanine 3.29-OSu (Cy3)—conjugated antibodies to anti-rabbit IgGs
(ab6939, Abcam) were used. For this analysis the cells were fixed by incubation with methanol for
20 min at –20 ◦C. Autophagy was detected also in viable cells using the “Autophagy Detection
Kit” (ab139484, Abcam) according to the manufacturer's protocol. Nuclei were stained with
4′-6-diamino-2-phenylindole dye (DAPI). DNA fragmentation was analyzed in paraformaldehyde-fixed cells by TUNEL assay using
“DeadEnd™Fluorometric TUNEL System” kit (G3250, Promega, Madison, WI, USA) according
to vendor’s instructions. HCV proteins were detected by indirect staining using monoclonal antibodies to the core (1F9),
NS3 (2H4), NS4 (a mixture of 3F12 to NS4A and 6B11 to NS4B proteins), and NS5A (3F4) proteins,
raised and described by us previously [63], as well as by commercially-available monoclonal antibodies
to E1 and E2 glycoproteins (using a mixture of ab21306 and ab20852, Abcam), or to the NS5B
protein (sc-58146, Santa Cruz Biotechnology, Dallas, TX, USA). As secondary antibodies, fluoresceine
isothiocianate-labeled anti-mouse IgGs (ab97039, Abcam) were used. In the DNA fragmentation assay
HCV proteins were stained using the primary antibodies listed above and secondary Alexa Fluor 594
(AF594)-labeled anti-mouse IgGs (R37121, Thermo Fisher Scientific). g
Simultaneous detection of HCV proteins and markers of apoptosis (caspases-3, -8, and -9)
was performed by double immunocytochemical staining with mixtures of the respective antibodies
and conjugates. Staining was visualized by the fluorescent microscope Axio Scope A1 Carl Zeiss (Germany) at
excitation/emission wavelengths of 520/560 nm (FITC), 552/565 nm (Cy3), 590/617 nm (AF594),
and 360/460 nm (DAPI). The stained cells were counted in no less than 8 different fields of view at
400× magnification by two independent observers. 4.7. Statistical Analysis Statistical analysis was performed with “Statistica 6” software (StatSoft Inc., Tulsa, OK, USA). All data on the graphs were presented as mean ratios of the number of stained cells to the total
number of cells ± standard error of means. The significance of differences between two groups were
determined using a Student’s t-test. A p-value < 0.05 was considered statistically significant. 4.6. Immunoblot Analysis The Western blot analysis was performed as described previously [43]. The primary antibodies
were used in 1% (w/v) non-fat milk in the following concentrations: anti-actin (Abcam, ab3280,
0.2 µg/mL), anti-NS3 (2H4, 1 µg/mL), anti-NS5A (3F4, 1.8 µg/mL), anti-NS5B (Santa-Cruz, sc-58146,
0.5 µg/mL). The HRP-conjugated anti-mouse (Santa-Cruz, sc-2005) secondary antibodies were used
at a concentration of 0.07 µg/mL. The signal was visualized using Pierce ECL Western Blotting Int. J. Mol. Sci. 2017, 18, 2346 12 of 16 substrate (Thermo Scientific, Waltham, MA, USA), with the exception of NS5B that was visualized
using SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Scientific). 5. Conclusions In conclusion, the data presented in this study point to different mechanisms by which HCV
proteins modulate cell death. For the first time, markers of apoptosis, necrosis, and autophagy
were analyzed simultaneously in hepatoma cells overexpressing individual virus proteins or their
combinations, or harboring the full-length replicon to ensure their more physiological expression in
the context of HCV genome replication. Most HCV proteins were shown to promote autophagy and to
exhibit different actions of apoptotic pathways. The structural core and E1/E2 proteins contributed
to apoptotic cell death, whereas the non-structural NS4A/B, NS5A, and NS5B proteins suppressed it
by blocking various members of the caspase cascade. A comparison of the viability of the transfected
cells with rates of detection of cell death markers suggests that death of NS3- and NS5B-expressing
cells occurs as a result of autophagy, and death of core-expressing cells occurs through the apoptotic
program, whereas E1/E2 glycoproteins favor both apoptotic and autophagy cell death. Finally, a stable
cell line harboring the full-length replicon exhibited strong signs of both apoptosis and autophagy. Our findings suggest possible molecular and cellular mechanisms that may present a target for the
treatment and/or prevention of virus-associated pathologies. Supplementary Materials: The following are available online at www.mdpi.com/1422-0067/18/11/2346/s1. Acknowledgments: Experiments with transient protein expression were supported by the Russian Foundation for
Basic Research (grant 17-04-01238), and studies with the HCV replicon system, by the Russian Science Foundation
(grant 14-14-01021). The authors are also grateful to Ralf Bartenschlager (University of Heidelberg) for his kind
gift of the HCV replicon system, and to Charles M. Rice (Rockefeller University) for Huh7.5 cells. Author Contributions: Olga V. Masalova, Vladimir S. Prassolov, Sergey N. Kochetkov, Alexander V. Ivanov,
and Alla A. Kushch conceived and designed the experiments; Olga V. Masalova, Ekaterina I. Lesnova,
Natalia F. Zakirova, and Alla A. Kushch performed the experiments; Olga V. Masalova, Ekaterina I. Lesnova,
Alexander V. Ivanov,
and Alla A. Kushch analyzed the data;
Vladimir S. Prassolov contributed
reagents/materials/analysis tools; Olga V. Masalova, Pavel N. Solyev, Sergey N. Kochetkov, Vladimir S. Prassolov,
Alexander V. Ivanov, and Alla A. Kushch wrote the paper. Conflicts of Interest: The authors declare no conflict of interest. of Interest: The authors declare no conflict of interes References 1. Dash, S.; Chava, S.; Aydin, Y.; Chandra, P.K.; Ferraris, P.; Chen, W.; Balart, L.A.; Wu, T.; Garry, R.F. Hepatitis c
virus infection induces autophagy as a prosurvival mechanism to alleviate hepatic er-stress response. Viruses
2016, 8. [CrossRef] [PubMed] 1. Dash, S.; Chava, S.; Aydin, Y.; Chandra, P.K.; Ferraris, P.; Chen, W.; Balart, L.A.; Wu, T.; Garry, R.F. Hepatitis c
virus infection induces autophagy as a prosurvival mechanism to alleviate hepatic er-stress response. Viruses
2016, 8. [CrossRef] [PubMed] 2. Ke, P.Y.; Chen, S.S. Hepatitis c virus and cellular stress response: Implications to molecular pathogenesis of
liver diseases. Viruses 2012, 4, 2251–2290. [CrossRef] [PubMed] 2. Ke, P.Y.; Chen, S.S. Hepatitis c virus and cellular stress response: Implications to molecular pathogenesis of
liver diseases. Viruses 2012, 4, 2251–2290. [CrossRef] [PubMed] 3. Wang, K. Autophagy and apoptosis in liver injury. Cell. Cycle 2015, 14, 1631–1642. [CrossRef] [PubM 3. Wang, K. Autophagy and apoptosis in liver injury. Cell. Cycle 2015, 14, 1631–1642. [CrossRef] [PubMed]
4. Hotchkiss, R.S.; Strasser, A.; McDunn, J.E.; Swanson, P.E. Cell death. N. Engl. J. Med. 2009, 361, 1570–1583. [CrossRef] [PubMed] 4. Hotchkiss, R.S.; Strasser, A.; McDunn, J.E.; Swanson, P.E. Cell death. N. Engl. J. Med. 2009, 361, 1570–1583. [CrossRef] [PubMed] 5. Hajnoczky, G.; Davies, E.; Madesh, M. Calcium signaling and apoptosis. Biochem. Biophys. Res. Commun. 2003, 304, 445–454. [CrossRef] 5. Hajnoczky, G.; Davies, E.; Madesh, M. Calcium signaling and apoptosis. Biochem. Biophys. Res. Commun. 2003, 304, 445–454. [CrossRef] 6. Ivanov, A.V.; Valuev-Elliston, V.T.; Tyurina, D.A.; Ivanova, O.N.; Kochetkov, S.N.; Bartosch, B.;
Isaguliants, M.G. Oxidative stress, a trigger of hepatitis c and b virus-induced liver carcinogenesis. Oncotarget
2017, 8, 3895–3932. [CrossRef] [PubMed] 6. Ivanov, A.V.; Valuev-Elliston, V.T.; Tyurina, D.A.; Ivanova, O.N.; Kochetkov, S.N.; Bartosch, B.;
Isaguliants, M.G. Oxidative stress, a trigger of hepatitis c and b virus-induced liver carcinogenesis. Oncotarget
2017, 8, 3895–3932. [CrossRef] [PubMed] Int. J. Mol. Sci. 2017, 18, 2346 13 of 16 7. Brenner, C.; Galluzzi, L.; Kepp, O.; Kroemer, G. Decoding cell death signals in liver inflammation. J. Hepatol. 2013, 59, 583–594. [CrossRef] [PubMed] 8. Alibek, K.; Irving, S.; Sautbayeva, Z.; Kakpenova, A.; Bekmurzayeva, A.; Baiken, Y.; Imangali, N.;
Shaimerdenova, M.; Mektepbayeva, D.; Balabiyev, A.; et al. Disruption of bcl-2 and bcl-xl by viral proteins
as a possible cause of cancer. Infect. Agent Cancer 2014, 9, 44. [CrossRef] [PubMed] 9. Jiang, X.; Kanda, T.; Wu, S.; Nakamoto, S.; Wakita, T.; Shirasawa, H.; Yokosuka, O. References Hepatitis c virus
nonstructural protein 5a inhibits thapsigargin-induced apoptosis. PLoS ONE 2014, 9, e113499. [CrossRef]
[PubMed] 10. Kong, L.; Li, S.; Huang, M.; Xiong, Y.; Zhang, Q.; Ye, L.; Liu, J.; Zhu, X.; Sun, R.; Guo, Y. The roles of
endoplasmic reticulum overload response induced by hcv and ns4b protein in human hepatocyte viability
and virus replication. PLoS ONE 2015, 10, e0123190. [CrossRef] [PubMed] ion. PLoS ONE 2015, 10, e0123190. [CrossRef] [PubMe 11. Aweya, J.J.; Sze, C.W.; Bayega, A.; Mohd-Ismail, N.K.; Deng, L.; Hotta, H.; Tan, Y.J. Ns5b induces
up-regulation of the bh3-only protein, bik, essential for the hepatitis c virus rna replication and viral
release. Virology 2015, 474, 41–51. [CrossRef] [PubMed] 12. Yeganeh, B.; Rezaei Moghadam, A.; Alizadeh, J.; Wiechec, E.; Alavian, S.M.; Hashemi, M.; Geramizadeh, B.;
Samali, A.; Bagheri Lankarani, K.; Post, M.; et al. Hepatitis b and c virus-induced hepatitis: Apoptosis,
autophagy, and unfolded protein response. World J. Gastroenterol. 2015, 21, 13225–13239. [CrossRef]
[PubMed] 13. Puri, P.; Chandra, A. Autophagy modulation as a potential therapeutic target for liver diseases. J. Clin. Exp. Hepatol. 2014, 4, 51–59. [CrossRef] [PubMed] 14. Sir, D.; Kuo, C.F.; Tian, Y.; Liu, H.M.; Huang, E.J.; Jung, J.U.; Machida, K.; Ou, J.H. Replication of hepatitis c
virus rna on autophagosomal membranes. J. Biol. Chem. 2012, 287, 18036–18043. [CrossRef] [PubMed] 15. Ke, P.Y.; Chen, S.S. Activation of the unfolded protein response and autophagy after hepatitis c virus infection
suppresses innate antiviral immunity in vitro. J. Clin. Investig. 2011, 121, 37–56. [CrossRef] [PubMed] 16. Shrivastava, S.; Raychoudhuri, A.; Steele, R.; Ray, R.; Ray, R.B. Knockdown of autophagy enhances the
innate immune response in hepatitis c virus-infected hepatocytes. Hepatology 2011, 53, 406–414. [CrossRef]
[PubMed] 17. Mohl, B.P.; Bartlett, C.; Mankouri, J.; Harris, M. Early events in the generation of autophagosomes are required
for the formation of membrane structures involved in hepatitis c virus genome replication. J. Gen. Virol. 2016, 97, 680–693. [CrossRef] [PubMed] 18. Shrivastava, S.; Bhanja Chowdhury, J.; Steele, R.; Ray, R.; Ray, R.B. Hepatitis c virus upregulates beclin1 for
induction of autophagy and activates mtor signaling. J. Virol. 2012, 86, 8705–8712. [CrossRef] [PubMed] 19. Medvedev, R.; Ploen, D.; Hildt, E. Hcv and oxidative stress: Implications for hcv life cycle and hcv-associated
pathogenesis. Oxid. Med. Cell. Longev. 2016, 2016, 9012580. [CrossRef] [PubMed] 20. Ploen, D.; Hildt, E. Hepatitis c virus comes for dinner: How the hepatitis c virus interferes with autophagy. World J. Gastroenterol. References 2015, 21, 8492–8507. [CrossRef] [PubMed] 21. Lim, E.J.; El Khobar, K.; Chin, R.; Earnest-Silveira, L.; Angus, P.W.; Bock, C.T.; Nachbur, U.; Silke, J.; Torresi, J. Hepatitis c virus-induced hepatocyte cell death and protection by inhibition of apoptosis. J. Gen. Virol. 2014,
95, 2204–2215. [CrossRef] [PubMed] 22. Ghosh Roy, S.; Sadigh, B.; Datan, E.; Lockshin, R.A.; Zakeri, Z. Regulation of cell survival and death during
flavivirus infections. World J. Biol. Chem. 2014, 5, 93–105. [CrossRef] [PubMed] 23. Masalova, O.V.; Lesnova, E.I.; Permyakova, K.Y.; Samokhvalov, E.I.; Ivanov, A.V.; Kochetkov, S.N.;
Kushch, A.A. Effect of hepatitis c virus proteins on the production of proinflammatory and profibrotic
cytokines in huh7.5 human hepatoma cells. Mol. Biol. (Mosk) 2016, 50, 422–430. [CrossRef] 24. Benali-Furet, N.L.; Chami, M.; Houel, L.; De Giorgi, F.; Vernejoul, F.; Lagorce, D.; Buscail, L.;
Bartenschlager, R.; Ichas, F.; Rizzuto, R.; et al. Hepatitis c virus core triggers apoptosis in liver cells
by inducing er stress and er calcium depletion. Oncogene 2005, 24, 4921–4933. [CrossRef] [PubMed] 25. Rajalakshmy, A.R.; Malathi, J.; Madhavan, H.N. Hcv core and ns3 proteins mediate toll like receptor induced
innate immune response in corneal epithelium. Exp. Eye Res. 2014, 128, 117–128. [CrossRef] [PubMed] 26. Soguero, C.; Joo, M.; Chianese-Bullock, K.A.; Nguyen, D.T.; Tung, K.; Hahn, Y.S. Hepatitis c virus core protein
leads to immune suppression and liver damage in a transgenic murine model. J. Virol. 2002, 76, 9345–9354. [CrossRef] [PubMed] Int. J. Mol. Sci. 2017, 18, 2346 14 of 16 14 of 16 27. Yang, S.H.; Lee, C.G.; Lee, C.W.; Choi, E.J.; Yoon, S.K.; Ahn, K.S.; Sung, Y.C. Hepatitis c virus core inhibits the
fas-mediated p38 mitogen activated kinase signaling pathway in hepatocytes. Mol. Cells 2002, 13, 452–462. [PubMed] 28. Chang, M.L.; Chen, J.C.; Chang, M.Y.; Yeh, C.T.; Lin, W.P.; Liang, C.K.; Huang, S.F.; Dang, K.N.; Chiu, C.T.;
Lin, D.Y. Acute expression of hepatitis c core protein in adult mouse liver: Mitochondrial stress and apoptosis. Scand. J. Gastroenterol. 2008, 43, 747–755. [CrossRef] [PubMed] 29. Ivanov, A.V.; Smirnova, O.A.; Petrushanko, I.Y.; Ivanova, O.N.; Karpenko, I.L.; Alekseeva, E.; Sominskaya, I.;
Makarov, A.A.; Bartosch, B.; Kochetkov, S.N.; et al. Hcv core protein uses multiple mechanisms to induce
oxidative stress in human hepatoma huh7 cells. Viruses 2015, 7, 2745–2770. [CrossRef] [PubMed] 30. Feng, S.; Li, M.; Zhang, J.; Liu, S.; Wang, Q.; Quan, M.; Zhang, M.; Cheng, J. References Regulation of hepg2 cell
apoptosis by hepatitis c virus (hcv) core protein via the sirt1-p53-bax pathway. Virus Genes 2015, 51, 338–346. [CrossRef] [PubMed] 31. Liu, C.; Qu, A.; Han, X.; Wang, Y. Hcv core protein represses the apoptosis and improves the autophagy of
human hepatocytes. Int. J. Clin. Exp. Med. 2015, 8, 15787–15793. [PubMed] 32. Galluzzi, L.; Bravo-San Pedro, J.M.; Vitale, I.; Aaronson, S.A.; Abrams, J.M.; Adam, D.; Alnemri, E.S.;
Altucci, L.; Andrews, D.; Annicchiarico-Petruzzelli, M.; et al. Essential versus accessory aspects of cell death:
Recommendations of the nccd 2015. Cell. Death Differ. 2015, 22, 58–73. [CrossRef] [PubMed] 33. Urbaczek, A.C.; Ribeiro, L.C.; Ximenes, V.F.; Afonso, A.; Nogueira, C.T.; Generoso, W.C.; Alberice, J.V.;
Rudnicki, M.; Ferrer, R.; Fonseca, L.M.; et al. Inflammatory response of endothelial cells to hepatitis c
virus recombinant envelope glycoprotein 2 protein exposure. Mem. Inst. Oswaldo Cruz 2014, 109, 748–756. [CrossRef] [PubMed] 34. Chiou, H.L.; Hsieh, Y.S.; Hsieh, M.R.; Chen, T.Y. Hcv e2 may induce apoptosis of huh-7 cells via a
mitochondrial-related caspase pathway. Biochem. Biophys. Res. Commun. 2006, 345, 453–458. [CrossRef]
[PubMed] 35. Zekri, A.N.; Sobhy, E.; Hussein, N.; Ahmed, O.S.; Hussein, A.; Shoman, S.; Soliman, A.H.; ElDin, H.M. E1/E2 of hepatitis c virus genotype4 and apoptosis. Asian Pac. J. Cancer Prev. 2016, 17, 3131–3138. [PubMed] 36. Hitomi, J.; Katayama, T.; Eguchi, Y.; Kudo, T.; Taniguchi, M.; Koyama, Y.; Manabe, T.; Yamagishi, S.;
Bando, Y.; Imaizumi, K.; et al. Involvement of caspase-4 in endoplasmic reticulum stress-induced apoptosis
and abeta-induced cell death. J. Cell. Biol. 2004, 165, 347–356. [CrossRef] [PubMed] 37. Szegezdi, E.; Fitzgerald, U.; Samali, A. Caspase-12 and er-stress-mediated apoptosis: The story so far. Ann. N. Y. Acad. Sci. 2003, 1010, 186–194. [CrossRef] [PubMed] 38. Chan, S.W.; Egan, P.A. Hepatitis c virus envelope proteins regulate chop via induction of the unfolded
protein response. FASEB J. 2005, 19, 1510–1512. [CrossRef] [PubMed] 39. Prikhod’ko, E.A.; Prikhod’ko, G.G.; Siegel, R.M.; Thompson, P.; Major, M.E.; Cohen, J.I. The ns3 protein of
hepatitis c virus induces caspase-8-mediated apoptosis independent of its protease or helicase activities. Virology 2004, 329, 53–67. [CrossRef] [PubMed] 40. Tanaka, M.; Nagano-Fujii, M.; Deng, L.; Ishido, S.; Sada, K.; Hotta, H. Single-point mutations of hepatitis c
virus ns3 that impair p53 interaction and anti-apoptotic activity of ns3. Biochem. Biophys. Res. Commun. 2006,
340, 792–799. [CrossRef] [PubMed] 41. Siu, G.K.; Zhou, F.; Yu, M.K.; Zhang, L.; Wang, T.; Liang, Y.; Chen, Y.; Chan, H.C.; Yu, S. References Hepatitis c virus
ns5a protein cooperates with phosphatidylinositol 4-kinase iiialpha to induce mitochondrial fragmentation. Sci. Rep. 2016, 6, 23464. [CrossRef] [PubMed] 42. Gong, G.; Waris, G.; Tanveer, R.; Siddiqui, A. Human hepatitis c virus ns5a protein alters intracellular
calcium levels, induces oxidative stress, and activates stat-3 and nf-kappa b. Proc. Natl. Acad. Sci. USA 2001,
98, 9599–9604. [CrossRef] [PubMed] 43. Ivanov, A.V.; Smirnova, O.A.; Ivanova, O.N.; Masalova, O.V.; Kochetkov, S.N.; Isaguliants, M.G. Hepatitis c
virus proteins activate nrf2/are pathway by distinct ros-dependent and independent mechanisms in huh7
cells. PLoS ONE 2011, 6, e24957. [CrossRef] [PubMed] 44. Smirnova, O.A.; Ivanova, O.N.; Bartosch, B.; Valuev-Elliston, V.T.; Mukhtarov, F.; Kochetkov, S.N.; Ivanov, A.V. Hepatitis c virus ns5a protein triggers oxidative stress by inducing nadph oxidases 1 and 4 and cytochrome
p450 2e1. Oxid. Med. Cell. Longev. 2016, 2016, 8341937. [CrossRef] [PubMed] Int. J. Mol. Sci. 2017, 18, 2346 15 of 16 15 of 16 45. Park, J.; Kang, W.; Ryu, S.W.; Kim, W.I.; Chang, D.Y.; Lee, D.H.; Park, D.Y.; Choi, Y.H.; Choi, K.; Shin, E.C.; et al. Hepatitis c virus infection enhances tnfalpha-induced cell death via suppression of nf-kappab. Hepatology
2012, 56, 831–840. [CrossRef] [PubMed] 46. Nomura-Takigawa, Y.; Nagano-Fujii, M.; Deng, L.; Kitazawa, S.; Ishido, S.; Sada, K.; Hotta, H. Non-structural protein 4a of hepatitis c virus accumulates on mitochondria and renders the cells prone to
undergoing mitochondria-mediated apoptosis. J. Gen. Virol. 2006, 87, 1935–1945. [CrossRef] [PubMed] 47. Targett-Adams, P.; Graham, E.J.; Middleton, J.; Palmer, A.; Shaw, S.M.; Lavender, H.; Brain, P.; Tran, T.D.;
Jones, L.H.; Wakenhut, F.; et al. Small molecules targeting hepatitis c virus-encoded ns5a cause subcellular
redistribution of their target: Insights into compound modes of action. J. Virol. 2011, 85, 6353–6368. [CrossRef]
[PubMed] 48. Silberstein, E.; Ulitzky, L.; Lima, L.A.; Cehan, N.; Teixeira-Carvalho, A.; Roingeard, P.; Taylor, D.R. Hcv-mediated apoptosis of hepatocytes in culture and viral pathogenesis. PLoS ONE 2016, 11, e0155708. [CrossRef] [PubMed] 49. Biederbick, A.; Kern, H.F.; Elsasser, H.P. Monodansylcadaverine (mdc) is a specific in vivo marker for
autophagic vacuoles. Eur. J. Cell. Biol. 1995, 66, 3–14. [PubMed] 50. Klionsky, D.J.; Cuervo, A.M.; Seglen, P.O. Methods for monitoring autophagy from yeast to human. Autophagy 2007, 3, 181–206. [CrossRef] [PubMed] 51. Chu, V.C.; Bhattacharya, S.; Nomoto, A.; Lin, J.; Zaidi, S.K.; Oberley, T.D.; Weinman, S.A.; Azhar, S.;
Huang, T.T. Persistent Expression of Hepatitis C Virus Non-Structural Proteins Leads to Increased Autophagy
and Mitochondrial Injury in Human Hepatoma Cells. References PLoS ONE 2011, 6, e28551. [CrossRef] [PubMed] 52. Quan, M.; Liu, S.; Li, G.; Wang, Q.; Zhang, J.; Zhang, M.; Li, M.; Gao, P.; Feng, S.; Cheng, J. A functional role
for ns5atp9 in the induction of hcv ns5a-mediated autophagy. J. Viral Hepat. 2014, 21, 405–415. [CrossRef]
[PubMed] 53. Su, W.C.; Chao, T.C.; Huang, Y.L.; Weng, S.C.; Jeng, K.S.; Lai, M.M. Rab5 and class iii phosphoinositide
3-kinase vps34 are involved in hepatitis c virus ns4b-induced autophagy. J. Virol. 2011, 85, 10561–10571. [CrossRef] [PubMed] 54. Wang, J.; Kang, R.; Huang, H.; Xi, X.; Wang, B.; Zhao, Z. Hepatitis c virus core protein activates autophagy
through eif2ak3 and atf6 upr pathway-mediated map1lc3b and atg12 expression. Autophagy 2014, 10, 766–784. [CrossRef] [PubMed] 55. Oberst, A.; Dillon, C.P.; Weinlich, R.; McCormick, L.L.; Fitzgerald, P.; Pop, C.; Hakem, R.; Salvesen, G.S.;
Green, D.R. Catalytic activity of the caspase-8-flip(l) complex inhibits ripk3-dependent necrosis. Nature 2011,
471, 363–367. [CrossRef] [PubMed] 56. Pietschmann, T.; Lohmann, V.; Rutter, G.; Kurpanek, K.; Bartenschlager, R. Characterization of Cell Lines
Carrying Self-Replicating Hepatitis C Virus RNAs. J. Virol. 2001, 75, 1252–1264. [CrossRef] [PubMed] 57. Walsh, M.J.; Vanags, D.M.; Clouston, A.D.; Richardson, M.M.; Purdie, D.M.; Jonsson, J.R.; Powell, E.E. Steatosis and Liver Cell Apoptosis in Chronic Hepatitis C: A Mechanism for Increased Liver Injury. Hepatology
2004, 39, 1230–1238. [CrossRef] [PubMed] 58. Blight, K.J.; McKeating, J.A.; Rice, C.M. Highly permissive cell lines for subgenomic and genomic hepatitis c
virus rna replication. J. Virol. 2002, 76, 13001–13014. [CrossRef] [PubMed] 59. Frese, M.; Barth, K.; Kaul, A.; Lohmann, V.; Schwarzle, V.; Bartenschlager, R. Hepatitis c virus rna replication
is resistant to tumour necrosis factor-alpha. J. Gen. Virol. 2003, 84, 1253–1259. [CrossRef] [PubMed] 60. Masalova, O.V.; Lesnova, E.I.; Grabovetskii, V.V.; Smirnova, O.A.; Ulanova, T.I.; Burkov, A.N.; Ivanov, A.V.;
Zaberezhnyi, A.D.; Ataullakhanov, R.I.; Kushch, A.A. DNA immunization with a plasmid carrying the gene
of hepatitis c virus protein 5a (ns5a) induces an effective cellular immune response. Mol. Biol. Mosk 2010, 44,
245–253. [CrossRef] 61. Masalova, O.V.; Lesnova, E.I.; Ivanov, A.V.; Pichugin, A.V.; Permiakova, K.; Smirnova, O.A.; Tynitskaia, V.L.;
Ulanova, T.I.; Burkov, A.N.; Kochetkov, S.N.; et al. Comparative analysis of the immune response to DNA
constructions encoding hepatitis c virus nonstructural proteins. Vopr. Virusol. 2013, 58, 21–28. [PubMed] 62. Permyakova, K.Y.; Lesnova, E.I.; Masalova, O.V.; Ivanov, A.V.; Ataullakhanov, R.I.; Kushch, A.A. Immunogenic properties of DNA constructions containing structural and nonstructural regions of hepatitis
c virus. Immunologiya 2015, 36, 162–167. References Int. J. Mol. Sci. 2017, 18, 2346 16 of 16 16 of 16 63. Rechkina, E.A.;
Denisova, G.F.;
Masalova, O.V.;
Lideman, L.F.;
Denisov, D.A.;
Lesnova, E.I.;
Ataullakhanov, R.I.; Gur’ianova, S.V.; Kushch, A.A. Mapping of antigenic determinants of hepatitis c
virus proteins using phage display. Mol. Biol. (Mosk) 2006, 40, 312–323. [CrossRef] 4. Perry, S.W.; Epstein, L.G.; Gelbard, H.A. Simultaneous in situ detection of apoptosis and necrosi
monolayer cultures by tunel and trypan blue staining. Biotechniques 1997, 22, 1102–1106. [PubMed] 64. Perry, S.W.; Epstein, L.G.; Gelbard, H.A. Simultaneous in situ detection of apoptosis and necrosis in
monolayer cultures by tunel and trypan blue staining. Biotechniques 1997, 22, 1102–1106. [PubMed]
65. Schlueter, N.; Lussi, A.; Ganss, C.; Gruber, R. L929 fibroblast bioassay on the in vitro toxicity of sncl2, h3po4,
clearfil se primer and combinations thereof. Swiss Dent. J. 2016, 126, 566–572. [PubMed] y
y
yp
g
q
65. Schlueter, N.; Lussi, A.; Ganss, C.; Gruber, R. L929 fibroblast bioassay on the in vitro toxicity of sncl2, h3po4,
clearfil se primer and combinations thereof. Swiss Dent. J. 2016, 126, 566–572. [PubMed] © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/).
|
Subsets and Splits
No community queries yet
The top public SQL queries from the community will appear here once available.