title
stringlengths 0
1.13k
| abstract
stringlengths 1
15.7k
| PMID
int64 22
36.5M
|
|---|---|---|
De novo adipogenesis for reconstructive surgery.
|
Autografting of lost soft tissue is an important subject of the plastic and reconstructive surgery and autograft of fat pads has been only technique for this goal. However, the results are disappointing because of absorption of the grafts with time. Adipoblasts or adipocyte precursor cells distribute widely in connective tissues and they can proliferate and mature into adipocytes even in the adult body. In experiments using mice, we found that de novo adipogenesis of endogenous precursor cells can be induced by injecting reconstituted basement membrane, Matrigel, supplemented with more than 1 ng/ml of bFGF. This adipogenesis was reproducibly induced by subcutaneous injection over the chest, lateral abdomen or head. Adipogenesis was induced even in ear cartilage or in muscle. To evaluate the possibility of future application of this de novo adipogenesis to plastic and reconstructive surgery, we have reviewed updated knowledge of the adipogenesis.
| 19,003,121
|
Effects of green, oolong and black teas and related components on the proliferation and invasion of hepatoma cells in culture.
|
The effects of teas and related components on the proliferation and invasion of cancer cells were examined by employing both in vitro proliferation and invasion assay systems. Powdered green, oolong and black tea extracts dose-dependently inhibited proliferation and invasion of a rat ascites hepatoma cell line of AH109A but did not affect the proliferation of the normal mesentery-derived mesothelial cells (M-cells) isolated from rats; higher concentrations of powdered oolong and black teas could restrain the proliferation of another tumor cell line of L929. The AH109A cells were found to penetrate underneath the monolayer of M-cells in the presence of 10% calf serum. When each rat serum obtained at 0.5, 1, 2, 3 and 5 h after oral intubation of each tea extract was added to the culture media instead of calf serum at a concentration of 10%, both the invasion and proliferation of AH109A were significantly suppressed. These ex vivo results suggest the potential bioavailability of effective tea components in rats. Furthermore, (-)-epigallocatechin gallate, (-)-epicatechin gallate and (-)-epigallocatechin from green tea as well as the mixture of theaflavin and theaflavin gallates from black tea were shown to be the most effective components against the invasion and proliferation of AH109A. These results show that the inhibitory effects of the teas and related components against AH109A cells are due to the cell-specific and higher sensitivity of the cell line to tea components.
| 19,003,122
|
Productivity enhancement of recombinant protein in CHO cells via specific promoter activation by oncogenes.
|
To construct a recombinant protein highly producing cell lines, we have previously developed the Oncogene Activated Production (OAP) system by using BHK-21 cells. Here we verified the availability of the OAP system in CHO cells. We firstly generated 'primed' ras amplified CHO cells, ras clone I, by introducing human c-Ha-ras oncogene into CHO cells. This ras clone I enables quick and easy establishment of recombinant protein hyper producing cell lines by introduction reporter gene of interest. Then we generated I13 by introducing human interleukin 6 (hIL-6) gene as a reporter gene, which showed enhanced productivity rate as compared to A7 established by conventional method. Furthermore, we found that hIL-6 production level of I13 was slightly improved by raising the CO(2) concentration from 5 to 8% possibly because of the enhanced growth rate. We further introduced the E1A oncogene, which has been shown to have a synergistic effect on the recombinant protein production of the ras-amplified BHK-21 cells, then evaluated the productivity. When culture in 5% CO(2) condition, only the slight effect can be seen. However when cultured in 8% CO(2) condition, not only cell number, but also productivity increased significantly, resulted in great augmentation of hIL-6 production, maximum production being 88.6 mug/ml/3 days. This study demonstrates that recombinant protein production level reached commercially desirable level by utilizing our OAP system in CHO cells and optimizing the culture condition.
| 19,003,130
|
A serum-free culture system for efficient in vitro production of bovine blastocysts with improved viability after freezing and thawing.
|
The aim of this study was to evaluate whether two completely serum-free media (IVMD101 and IVD101) could improve the yield and quality of bovine blastocysts from in vitro matured and fertilized oocytes. The media were evaluated in the presence (IVMD101) or absence (IVD101) of bovine cumulus/granulosa cell (BCGC) cocultures. The proportion of embryos developing to the blastocyst stage in IVMD101 medium with BCGC cocultures (36.5%) and IVD101 medium without BCGC cocultures (37.1%) was significantly higher than in serum-supplemented medium (TCM199 + 5% calf serum) with BCGC cocultures (25.1%). Furthermore, the mean cell numbers per blastocyst on Day 7 developed in IVMD101 medium (179.5 cells) and IVD101 medium (177.1 cells) were greater than in the serum-supplemented medium (145.7 cells). The survival rates of blastocysts derived in IVMD101 medium (73.3%) and IVD101 medium (60.0%) based on hatching after 72 h of post-thaw culture were superior to that of blastocysts derived in the serum-supplemented medium (48.1%). Under microscopic observation, bovine blastocysts derived in the serum-supplemented medium showed abundant lipid droplets, largely into the trophectoderm cells. This morphological difference may partly explain the sensitivity of serum-derived embryos after freezing and thawing. In conclusion, these new serum-free culture media are useful, not only to study the mechanisms of early embryogenesis, but also for mass production of good quality embryos for embryo transfer, cloning and transgenesis.
| 19,003,132
|
Co-expression of bcl-2 and bag-1, apoptosis suppressing genes, prolonged viable culture period of hybridoma and enhanced antibody production.
|
Human bcl-2 and bag-1 DNA were introduced into mouse hybridoma 2E3- O cells and expressed. The expression of bcl-2 in BCMGneo-bcl2 transfectants was confirmed by ELISA and that of bag-1 in pZeo-bag1 was confirmed by western blotting. In batch cultures, the over-expression of bcl-2 prolonged the culture period by 2 days and co-expression of bcl-2 and bag-1 prolonged the culture period by 3 days. The delayed increase in the dead cell number in culture of the bcl-2 and bag-1 cotransfectant indicated the additional antiapoptosis effect of bcl-2 and bag-1 cotransfection in comparison with the bcl-2 only transfection. The bcl-2 transfectants (2E3O-Bcl2) produced antibody twofold per batch culture in comparison with 2E3-O cells transfected with BCMGSneo (2E3O-Mock). Enhancement of this MoAb production was due to the improved survival of the cells and was not due to stimulation of antibody production rate per cell by Bcl-2 expression. And the bcl-2 and bag-1 co-transfectant (2E3O-Bcl2-BAG1) produced antibody approximately fourfold of 2E3O-Mock per batch culture. Enhancement of this MoAb production was due to the improved survival of the cells and was partly due to stimulation of MoAb production rate per cell in the non-growing phase by the cotransfection. The method to engineer hybridoma cells genetically with bcl-2 and bag-1 for increasing viability and productivity would be widely applied for improving antibody productivity of hybridoma cultures.
| 19,003,134
|
The neurite-initiating effect of microbial extracellular glycolipids in PC12 cells.
|
The effects of several kinds of microbial extracellular glycolipids on neurite initiation in PC12 cells were examined. Addition of mannosylerythritol lipid-A (MEL-A), MEL-B, and sophorose lipid (SL) to PC12 cells caused significant neurite outgrowth. Other glycolipids, such as polyol lipid (PL), rhamnose lipid (RL), succinoyl trehalose lipid-A (STL-A) and STL-B caused no neurite-initiation. MEL-A increased acetylcholine esterase (AChE) activity to an extent similar to nerve growth factor (NGF). However, MEL-A induced one or two long neurites from the cell body, while NGF induced many neurites. In addition, MEL-A-induced differentiation was transient, and after 48 h, percentage of cells with neurites started to decrease in contrast to neurons induced by NGF, which occurred in a time-dependent manner. MEL-A could induce neurite outgrowth after treatment of PC12 cells with an anti-NGF receptor antibody that obstructed NGF action. These results indicate that MEL-A and NGF induce differentiation of PC12 cells through different mechanisms.
| 19,003,137
|
Modulation by extracellular matrices of monooxygenase and CYP1A1 induction in Hep G2 cells in serum-free culture.
|
The in vitro cellular functions of differentiated cells are influenced by culture conditions. Effects of several extracellular matrices (ECMs) on cytochrome P450-dependent monooxygenases (MFOs) induction and cytochrome P4501A1 (CYP1A1) gene expression were estimated in Hep G2 cells cultured in a serum-free medium. The cells were cultured on collagen type I- and II-, fibronectin-, and matrigel-coated dishes and MFO activities were induced by the addition of 3-methylcholanthrene (MC). The induction of ethoxycoumarin O-deethylase (ECOD) and alkoxyresorufin O-dealkylase activities as well as the expression of CYP1A1 mRNA were also determined. ECOD and methoxy- and ethoxyresorufin O-dealkylase activities in Hep G2 cells were enhanced by culturing the cells using a serum-free medium on fibronectin- or matrigel-coated dishes. ECOD activity on fibronectin-coated dishes was about 3-fold higher than that using a serum-supplemented medium on untreated dishes. Furthermore, both immobilized and soluble fibronectin enhanced the induction of MFOs. The expression of CYP1A1 mRNA using fibronectin-coated dishes was about 2-fold higher than that using a serum-supplemented medium on untreated dishes. These findings suggest that the gene expression in cultured cells is greatly influenced by ECMs. By using fibronectin-coated dishes to cell culture in a serum-free medium, reproducible and highly sensitive results can be obtained in experiments using cultured cells.
| 19,003,142
|
Analysis of the use of fortified medium in continuous culture of mammalian cells.
|
Continuous culture is frequently used in the cultivation of mammalian cells for the manufacturing of recombinant protein pharmaceuticals. In such operations a large volume of medium is turned over each day, especially in the case where cell recycle, or perfusion cultivation, is practiced. In principle, the volumetric throughput of medium can be reduced by using a more concentrated feed while maintaining the same nutrient provision rate. Overall, the medium components are divided into two categories: 'consumable nutrients' and 'unconsumable inorganic bulk salts'. In such fortified medium, the concentrations of consumable nutrients, but not bulk salts, are increased. With a stoichiometrically-balanced medium, the large amount of nutrients fed into the culture is largely consumed by cells to give rise to residual concentrations of these nutrients in their optimal range. However, unless care is taken to initiate the continuous culture, overshoot of nutrients may occur during the transient period. The high nutrient concentration during overshoot may be inhibitory by itself, or the resulting high osmolality may retard the growth. Using a mathematical model that incorporates the growth inhibitory effect of high osmolality we demonstrate such a potentially catastrophic effect of nutrient and osmolality overshoot by simulation. To avoid overshoot a controlled nutrient feeding scheme should be devised at the initiation of continuous culture.
| 19,003,148
|
Development of an in vitro system for screening the ligands of a membrane glycoprotein CD36.
|
It has well been known that human and rodents exhibit a preference for fats. This suggests the existence of an orosensory system responsible for the detection of dietary fats. A plasma membrane glycoprotein CD36, besides the role in the uptake of long-chain fatty acids (LCFAs) as well as oxidized low-density lipoprotein (OxLDL) in a variety of cells, has been postulated to be a candidate fat taste receptor on the tongue. Therefore, molecules that bind with CD36 to cause intracellular signaling but have fewer calories could be substitutes for dietary fats. In the present study, we developed an in vitro system for the screening of CD36 ligands using Chinese hamster ovary-K1 cells (CHO-K1) stably transfected with human or mouse CD36. When incubated with OxLDL labeled with fluorescence dye, the fluorescence was much higher in the transfected CHO-K1 cells than in non-transfected CHO-K1 cells. Incubation of the transfected cells with OxLDL caused a rapid phosphorylation of extracellular signal regulated kinase, and the degree was significantly higher compared with that in non-transfected CHO-K1 cells. The expression system using CHO-K1 cells could be a convenient tool to screen the novel ligands of CD36.
| 19,003,159
|
Cephalic hedgehog expression is regulated directly by Sox17 in endoderm development of Xenopus laevis.
|
In early development of animals, hedgehog (Hh) genes function as morphogen in the axis determination and the organ formation. In Xenopus, three hedgehog genes, sonic (shh), banded (bhh), and cephalic (chh), were identified and might organize various tissues and organs in embryogenesis. Here, we report the spatial and temporal regulation of Xchh which is expressed in endoderm cells differentiating to digestive organs. Xchh expression in endoderm was inhibited by ectopic expression of the dominant-negative activin receptor, tAR. Moreover, a maternally inherited transcription factor VegT and its downstream regulators activated Xchh expression. These indicates that Xchh is regulated by the factor involved in the cascade originated from VegT via activin/nodal signals. Using the Sox17alpha-VP16-GR construct, we showed that Xchh expression might be induced directly by transcription factor Sox17.
| 19,003,160
|
A single plasmid transfection that offers a significant advantage associated with puromycin selection in Drosophila Schneider S2 cells expressing heterologous proteins.
|
The Drosophila Schneider S2 (S2) Expression System enables expression of recombinant proteins constitutively, as well as inductively. This system can establish both transient and stable transformants with various selection markers. The generation of stable cell lines for increased expression or large scale expression of the desired protein is currently accomplished by cotransfection of both the expression and selection vectors. The selection vectors, pCoHYGRO and pCoBLAST, are commercially available using hygromycin-B and blasticidin S, respectively. Recently, we generated a plasmid, pCoPURO, for selection of transfected S2 cells using puromycin, which allows significant acceleration of the selection time. Although co-transfection of the selection marker with the plasmid for heterologous protein expression is functional in stable expression at short culture periods, the expression levels of stable transformants are continuously decreased during long culture times. To overcome this limitation, we generated pMT-PURO, a new plasmid that contains both the expression cassette and puromycin selection marker in a single plasmid. This system allows rapid selection and maintenance of the transformed S2 lines for extended culture periods.
| 19,003,171
|
Bioethics and biotechnology.
|
Biotechnology is at the intersection of science and ethics. Technological developments are shaped by an ethical vision, which in turn is shaped by available technology. Much in biotechnology can be celebrated for how it benefits humanity. But technology can have a darker side. Biotechnology can produce unanticipated consequences that cause harm or dehumanise people. The ethical implications of proposed developments must be carefully examined. The ethical assessment of new technologies, including biotechnology, requires a different approach to ethics. Changes are necessary because new technology can have a more profound impact on the world; because of limitations with a rights-based approach to ethics; because of the importance and difficulty of predicting consequences; and because biotechnology now manipulates humans themselves. The ethical questions raised by biotechnology are of a very different nature. Given the potential to profoundly change the future course of humanity, such questions require careful consideration. Rather than focussing on rights and freedoms, wisdom is needed to articulate our responsibilities towards nature and others, including future generations. The power and potential of biotechnology demands caution to ensure ethical progress.
| 19,003,197
|
Immuno-chromatographic assay for diagnosis of feline leukemia virus infection.
|
Feline leukemia virus (FeLV) infectious disease is one of feline infection diseases spreading broadly all over the world. For bedside diagnosis of FeLV infectious disease, an immuno-chromatographic assay was investigated. Five different monoclonal antibodies were developed against the major core protein FeLV-p27. Among them, the combination of FL6 and FL12, which had little epitopic overlap each other, showed the highest sensitivity with no cross-reaction to the other feline virus antigens when they were employed to the immuno-chromatographic assay. The system had a practical detection limit of 0.5 ng of FeLV-p27 per 0.1 ml of feline sera within 15 min. In comparison with clinical standard methods, the system gave rapidly and accurately the same diagnosis with neither false negative nor false positive. Moreover, it did not need any pretreatment of blood specimen.
| 19,003,209
|
Development of monoclonal antibody against isoquinoline alkaloid coptisine and its application for the screening of medicinal plants.
|
In the development of immunoassay technique, the design of hapten containing a functional group suitable for protein conjugate is the key step for the preparation of antibodies against small molecules. Coptisine (MW 320), a bioactive constituent of Berberis and Coptis species, is small as an immunogen. In addition, coptisine has no reactive group in molecule for conjugating with a protein. To overcome this problem, 9-O-carboxymethyl-berberrubine was designed and conjugated with carrier protein. In order to confirm its immunogenicity, the ratio of hapten in the conjugate was determined by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). After immunization, hybridomas secreting antibodies against coptisine were produced by fusing splenocytes with mouse myeloma cell line, P3-X63-Ag8-653. Among hybridomas, the clone 2A1 secreting anti-coptisine monoclonal antibody (MAb) 2A1-9E-1 was obtained through the limited dilution method. The MAb-based enzyme-linked immunosorbent assay (ELISA) against coptisine was developed and characterized. The linear range of the assay in this ELISA method was extended from 1.56 to 25 mug ml(-1) possessing the detection limit of 1.56 mug ml(-1). The established ELISA using MAb 2A1-9E-1 was applied for the survey of isoquinoline alkaloids in various medicinal plants.
| 19,003,234
|
The use of hTERT-immortalized cells in tissue engineering.
|
The use of human telomerase reverse transcriptase (hTERT)-immortalized cells in tissue engineering protocols is a potentially important application of telomere biology. Several human cell types have been created that overexpress the hTERT gene with enhanced telomerase activity, extended life span and maintained or even improved functional activities. Furthermore, some studies have employed the telomerized cells in tissue engineering protocols with very good results. However, high telomerase activity allows extensive cell proliferation that may be associated with genomic instability and risk for cell transformation. Thus, safety issues should be studied carefully before using the telomerized tissues in the clinic. Alternatively, the development of conditional or intermittent telomerase activation protocols is needed.
| 19,003,242
|
Microplates with integrated oxygen sensing for medium optimization in animal cell culture.
|
A new approach using microtiter plate cultivation with on-line measurement of dissolved oxygen (DO) was applied for medium optimization of mammalian cell culture. Applying dynamic liquid phase balance, oxygen uptake rates were calculated from the DO level and used as an indicator for culture viability. The developed method was successfully applied to optimization of the concentration of glucose, glutamine and inorganic salts for cultivation of a Chinese Hamster Ovary (CHO) cell line. Using 2(3) full factorial central composite design, the optimum medium composition could be identified in one single run. The concentration of inorganic salts had a significant influence on cultivation. The developed method exhibits high potential to improve procedures of medium optimization for animal cell cultivation by allowing the investigation of large sets of potentially important variables in short time and with reduced effort.
| 19,003,253
|
Preparation of secretory vesicle-free plasma membranes by isopycnic sucrose gradient fractionation of neutrophils purified by the gelatin method.
|
Isolated human neutrophils serve as a model for the in vitro study of host defensive processes as well as the cell biology and biochemistry of primary human cells. We demonstrate that the requirements of the gelatinbased procedure for neutrophil isolation from whole blood induces the complete loss of secretory vesicles from in vitro isolated populations, whereas isolation by a dextran-based methodology results in the preservation of this organelle. Following density fractionation of cellular cavitates, examination of commonly employed plasma membrane marker activities yielded subcellular localization patterns that were indistinguishable between dextran- or gelatin-isolated populations, indicating both populations to be otherwise comparable in terms of the relative complexity and large-scale organization of plasma membranes. Given that the cell surface upregulation of secretory vesicles is implicated as an initial requirement of neutrophil activation as well as an intrinsic feature of neutrophil priming, we show that dextran and gelatin-isolated neutrophils may be considered to occupy functionally nonactivated and primed cellular states, respectively. These differences in phenotype can be exploited in specific ways. We suggest that the gelatin method has technical advantages with regard to the study of neutrophil plasma membranes. In particular, results from this study indicate the gelatin method to be a reliable and effective preparatory technique appropriate for tandem use with density fractionation procedures to achieve rapid isolation of plasma membranes that are uncontaminated by secretory organelles.
| 19,003,266
|
Primary adipocyte culture: adipocyte purification methods may lead to a new understanding of adipose tissue growth and development.
|
In the present manuscript, the methods required to generate purified cultures of mature adipocytes, as well as stromal vascular cells, from the same isolation are detailed. Also, we describe the in vitro conditions for the dedifferentiation of the isolated mature adipocytes. These two types of cells may be used to reevaluate differences between presently available cellular models for lipogenesis/lipolysis and might provide a new cellular physiological system for studies utilizing the proliferative progeny from mature adipocyte dedifferentiation. Alternative possibilities to the dedifferentiation phenomenon are proposed, as this new area of research is novel.
| 19,003,270
|
CB.Hep-1 hybridoma growth and antibody production using protein-free medium in a hollow fiber bioreactor.
|
The protein-free medium TurboDoma HP.1 (THP.1) was used to produce the CB.Hep-1 monoclonal antibody (mAb) in a CP-1000 hollow fiber bioreactor (HFB). This mAb is used for the immunopurification of recombinant hepatitis B surface antigen (rHBsAg), which is included in a vaccine preparation against the Hepatitis B Virus. By using the experimental conditions tested in this work we were able to generate more than 433 mg of IgG in 43 days. The maximum antibody concentration obtained was about 2.4 mg ml(-1)and the IgG production per day was approximately 11 mg of monoclonal antibody, which constitutes a good concentration value in comparison to the results obtained in ascitic fluid, where concentration for this hybridoma was around 3 mg ml(-1). We used different analytical methods to control the quality of mAbs, obtained from the in vitro system. They included affinity constant determination, analysis of N-glycan structures, immunoaffinity chromatography and antigen binding properties. The results obtained suggest that no significant changes occurred in the mean characteristics of the mAb harvested from the bioreactor during the 43 days of cultivation.
| 19,003,291
|
Epitope analysis of human monoclonal antibody specific for rice allergenic protein generated by in vitro immunization.
|
We previously established an in vitro immunization protocol for generating antigen specific human monoclonal antibodies (mAbs). In vitro immunization was performed against the soluble protein of rice allergenic protein (RA), resulting in the generation of three B cell clones, AC7-1/F9, CB7-1/E2 and CB7-8/F5, all of which produce a RA-specific human monoclonal IgM antibody. We attempted to map the epitope regions recognized by thesem Abs to characterize their specificities. We performed two rounds of epitope mapping, rough mapping using 10-mer peptides covering the full-length RA with 5 amino acids overlapping, and fine mapping using 8-mer peptides covering the putative epitope regions from the rough mapping with 1amino acid overlapping. As a result of the fine mapping,we identified the epitope regions of these three mAbs as(45)QVWQDCCRQ(54)L, (56)AVDDGWCRCGA(67)L and(91)FPGCRRG(98)D on the RA molecule and found to be identical. Furthermore, we determined the putative core epitope regions, which are critical for mAb binding to each region, (47)WQDCC(52)R and (60)GWC(63)R. The information about the epitope region on the RA molecule,which might trigger the allergenic response, would be useful to establish a specific immunotherapy against rice allergy.
| 19,003,321
|
Preparation of monoclonal antibody against crocin and its characterization.
|
Three crocin-carrier protein conjugates were synthesized and their hapten numbers were determined by matrix-assisted laser desorption/ionization time of flight mass spectrometry. Three monoclonal antibodies against crocin were produced by hybridomas fused with the splenocytes immunized with crocin hemisuccinate-bovine serum albumin conjugate and HAT-sensitive mouse myeloma cell line, P3-X63-Ag8-653. They were identified as IgG2a and IgG2b possessing lambda light chain, respectively. Their wide reactivities against crocetin glycosides were discussed.
| 19,003,338
|
Hybridoma cell behaviour in continuous culture under hyperosmotic stress.
|
In this paper, we propose an alternative strategy to the ones proposed before (Oh et al., 1993; Øyaas et al., 1994a) to get real increases of global final antibody titer and production at hyperosmotic stress, by reducing the detrimental effect of such a stress on cell growth, and conserving the stimulating effect on antibody production. It consists of cultivating the cells in continuous culture and increasing the osmolality stepwise. In this way, the cells could progressively adapt to the higher osmolality at each step and antibody titers could be nearly doubled at 370 and 400 mOsm kg-1, compared to the standard osmolality of 335 mOsm kg-1. Surprisingly, the stimulation of antibody production was not confirmed for higher osmolalities, 425 and 450 mOsm kg- 1, despite the minor negative effect on cell growth. Intracellular IgG analysis by flow cytometry revealed at these osmolalities a significant population of non-producing cells. However, even when taking into account this non-producing population, a stimulating effect on antibody production could not be shown at these highest osmolalities. It seems to us that osmolality has a significant effect on the appearance of these non-producing cells, since they were not observed in continuous cultures at standard osmolality, of comparable duration and at an even higher dilution rate. The appearance of the non-producing cells coincides furthermore with modifications of the synthesised antibody, as shown by electrophoretic techniques. It is however not really clear if these two observations reflect actually the same phenomenon. Hyperosmolality affects the cell behaviour in continuous culture in multiple ways, independently of the growth rate, counting all at least partially for the observed stimulation of antibody production: acceleration of the amino acid, and in particular the glutamine metabolism, increase of the cell volume, increase of the intracellular pH and accumulation of cells in the G1 cell cycle phase.
| 19,003,339
|
Construction and characterization of stably transfected BHK-21 cells with human-type sialylation characteristic.
|
The human Golgi enzyme CMP-NeuAc:Gal(beta1-4)GlcNAc-R alpha2,6-sialyltransferase (ST6N) was stably coexpressed with human erythropoietin (EPO) from a BHK-21A cell line. The cell line was characterized with respect to the expression and in vitro activity of the ST6N and the endogenous alpha2,3-sialyltransferase. Detailed structural analysis of the N-linked carbohydrates of the rhuEPO expressed from the new cell line was performed by HPAE-PAD-mapping, MALDI/TOF-MS and methylation analysis after purification of the recombinant protein by immunoaffinity chromatography. This is the first report describing that the human alpha2,6-sialyltransferase is capable of sialylating, apart from Gal(beta1-4)GlcNAc-R, also GalNAc(beta1-4)GlcNAc-R motifs in vivo, which is not the case for the endogenous BHK-cell alpha2,3-sialyltransferase.
| 19,003,352
|
Influence of bcl-2 on antibody productivity in high cell density perfusion cultures of hybridoma.
|
Apoptosis is an active, genetically determined death mechanism which can be induced by a wide range of physiological factors and by mild stress. It is the predominant form of cell death during the production of antibodies from murine hybridoma cell lines. A number of studies have now demonstrated that the suppression of this death pathway, by means of over-expression of survival genes such as bcl-2, results in improved cellular robustness and antibody productivity during batch culture. In the present study, the influence of bcl-2 expression on hybridoma productivity in two high density perfusion bioreactor systems was investigated. In the first system, a fixed-bed reactor, the DNA content in the spent medium was 25% higher in the control (TB/C3-pEF) culture than that found in the bcl-2 transfected (TB/C3-bcl2) cultures at all perfusion rates. This is indicative of a higher level of cell death in the control cell line. The average antibody concentration for the TB/C3-pEF cell line was 14.9 mg L-1 at perfusion rates of 2.6 and 5.2 d-1. However, for the TB/C3-bcl2 cell line it was 33 mg L-1 at dilution rates of 2 and 4 d-1. A substantial increase in antibody concentration was also found in the Integra Tecnomouse hollow fibre reactor. The antibody titre in the TB/C3-bcl2 cassette was nearly 100% higher than that in the TB/C3-pEF cassette during the cultivation period which lasted 6 weeks. Clearly, these results demonstrate the positive impact of bcl-2 over-expression on production of antibody in hybridoma perfusion cultures.
| 19,003,359
|
Cell engineering for muscle gene therapy: Extemporaneous production of retroviral vector packaging macrophages using defective herpes simplex virus type 1 vectors harbouring gag, pol, env genes.
|
Gene therapy as a treatment for neuromuscular diseases is an ever-developing concept based on the use of DNA as the therapeutic agent. In the search for appropriate strategies a bottleneck exists, however, concerning the targeting of vectors carrying the therapeutic gene, to all pathologic sites. These diseases are often characterised by multiple widespread lesions spread over a large area, rendering administration by local injection into tissues, clinically irrelevant. With this in mind, we have proposed that circulating cells (monocytes/macrophages), which home naturally to inflammatory lesions, characteristic of degenerating muscle, could be used as shuttles able to track down every damaged site, and deliver there a corrective gene. Our aim is to mobilise a corrective gene from these infiltrating monocyte-macrophages, into muscle cells, a process of in situ cell to cell gene transfer which could be accomplished using a retroviral vector, since the regeneration process involves the proliferation of muscle precursors before they fuse to form replacement fibres. For this, monocyte-macrophages must be engineered into 'packaging cells' containing both the replication deficient retrovirus carrying the gene of interest and an helper genome (gag-pol-env) needed for its assembly and secretion. Here, we have transduced a monocyte cell line using a defective murine Moloney leukemia retrovirus carrying the LacZ reporter gene. This provided us with a platform to investigate the possibility of gag-pol-env vector driven packaging of the defective retrovirus by macrophages. We show that an herpes simplex virus type I amplicon harbouring the Moloney gag, pol, env sequences is able to rescue the defective retrovirus vector from macrophages, allowing gene transfer into muscle precursor cells. After fusion, these cells gave rise to genetically modified myotubes in vitro.
| 19,003,367
|
Justification of continuous packed-bed reactor for retroviral vector production from amphotropic PsiCRIP murine producer cell.
|
To indentify a plausible large-scale production system forretroviral vector, three culture systems, i.e., batch culturewith medium exchange, microcarrier culture, and packed-bedreactor culture were compared. In batch cultures with mediumexchange, high cell concentrations were maintained for about amonth, and the harvested retroviral titer remained constant. Inmicrocarrier cultures, although cell growth was rapid, theretroviral titer was unexpectedly low, suggesting that the lowtiter was due either to serious damage to the retroviral vectoror to a reduction in the production rate of retroviral vector,caused by mechanical shear forces. Although the retroviral titer(maximum titer, 1.56 x 10(6)) in the packed-bedreactor was a little bit lower than that obtained in the batchculture with medium exchange (maximum titer, 1.91 x10(6)), continuous production made it possible to increasethe cumulative titer up to 16-fold of that from the batchculture with medium exchange. Moreover, as the packed-bedreactor system requires less labor and shows excellentvolumetric productivity in comparison to batch cultures withmedium exchanges, it will be an appropriate production systemfor retroviral vector in large quantities.
| 19,003,389
|
Isolation, characterization and recombinant protein expression in Veggie-CHO: A serum-free CHO host cell line.
|
The dihydrofolate reductase-deficient Chinese hamster ovary cell line, DXB11-CHO, commonly used as a host cell for the production of recombinant proteins requires 7.5% serum-supplementation for optimal growth. Regulatory issues surrounding the use of serum in clinical production processes and the direct and indirect costs of using serum in large-scale production and recovery processes have triggered efforts to derive serum-independent host cell lines. We have successfully isolated a serum-free host that we named Veggie- CHO. Veggie-CHO was generated by adapting DXB11-CHO cells to growth in serum-free media in the absence of exogenous growth factors such as Transferrin and Insulin-like growth factor, which we have previously shown to be essential for growth and viability of DXB11- CHO cells. Veggie-CHO cells have been shown to maintain an average doubling time of 22 hr in continuous growth cultures over a period of three months and have retained the dihydrofolate reductase -deficient phenotype of their parental DXB11-CHO cells. These properties and the stability of its serum-free phenotype have allowed the use of Veggie- CHO as host cells for transfection and amplified expression of recombinant proteins. We describe the derivation a serum-free recombinant cell line with an average doubling time of 20 hr and specific productivity of 2.5 Units recombinant Flt-3L protein per 10e6 cells per day.
| 19,003,404
|
Systems biology via redescription and ontologies (I): finding phase changes with applications to malaria temporal data.
|
Biological systems are complex and often composed of many subtly interacting components. Furthermore, such systems evolve through time and, as the underlying biology executes its genetic program, the relationships between components change and undergo dynamic reorganization. Characterizing these relationships precisely is a challenging task, but one that must be undertaken if we are to understand these systems in sufficient detail. One set of tools that may prove useful are the formal principles of model building and checking, which could allow the biologist to frame these inherently temporal questions in a sufficiently rigorous framework. In response to these challenges, GOALIE (Gene ontology algorithmic logic and information extractor) was developed and has been successfully employed in the analysis of high throughput biological data (e.g. time-course gene-expression microarray data and neural spike train recordings). The method has applications to a wide variety of temporal data, indeed any data for which there exist ontological descriptions. This paper describes the algorithms behind GOALIE and its use in the study of the Intraerythrocytic Developmental Cycle (IDC) of Plasmodium falciparum, the parasite responsible for a deadly form of chloroquine resistant malaria. We focus in particular on the problem of finding phase changes, times of reorganization of transcriptional control.
| 19,003,444
|
Modelling molecular interaction pathways using a two-stage identification algorithm.
|
In systems biology, molecular interactions are typically modelled using white-box methods, usually based on mass action kinetics. Unfortunately, problems with dimensionality can arise when the number of molecular species in the system is very large, which makes the system modelling and behavior simulation extremely difficult or computationally too expensive. As an alternative, this paper investigates the identification of two molecular interaction pathways using a black-box approach. This type of method creates a simple linear-in-the-parameters model using regression of data, where the output of the model at any time is a function of previous system states of interest. One of the main objectives in building black-box models is to produce an optimal sparse nonlinear one to effectively represent the system behavior. In this paper, it is achieved by applying an efficient iterative approach, where the terms in the regression model are selected and refined using a forward and backward subset selection algorithm. The method is applied to model identification for the MAPK signal transduction pathway and the Brusselator using noisy data of different sizes. Simulation results confirm the efficacy of the black-box modelling method which offers an alternative to the computationally expensive conventional approach.
| 19,003,449
|
Engineering-approach accelerates computational understanding of V1-V2 neural properties.
|
We present two computational models (i) long-range horizontal connections and the nonlinear effect in V1 and (ii) the filling-in process at the blind spot. Both models are obtained deductively from standard regularization theory to show that physiological evidence of V1 and V2 neural properties is essential for efficient image processing. We stress that the engineering approach should be imported to understand visual systems computationally, even though this approach usually ignores physiological evidence and the target is neither neurons nor the brain.
| 19,003,454
|
A retinal circuit model accounting for wide-field amacrine cells.
|
In previous experimental studies on the visual processing in vertebrates, higher-order visual functions such as the object segregation from background were found even in the retinal stage. Previously, the "linear-nonlinear" (LN) cascade models have been applied to the retinal circuit, and succeeded to describe the input-output dynamics for certain parts of the circuit, e.g., the receptive field of the outer retinal neurons. And recently, some abstract models composed of LN cascades as the circuit elements could explain the higher-order retinal functions. However, in such a model, each class of retinal neurons is mostly omitted and thus, how those neurons play roles in the visual computations cannot be explored. Here, we present a spatio-temporal computational model of the vertebrate retina, based on the response function for each class of retinal neurons and on the anatomical inter-cellular connections. This model was capable of not only reproducing the spatio-temporal filtering properties of the outer retinal neurons, but also realizing the object segregation mechanism in the inner retinal circuit involving the "wide-field" amacrine cells. Moreover, the first-order Wiener kernels calculated for the neurons in our model showed a reasonable fit to the kernels previously measured in the real retinal neuron in situ.
| 19,003,460
|
Outline of a novel architecture for cortical computation.
|
In this paper a novel architecture for cortical computation has been proposed. This architecture is composed of computing paths consisting of neurons and synapses. These paths have been decomposed into lateral, longitudinal and vertical components. Cortical computation has then been decomposed into lateral computation (LaC), longitudinal computation (LoC) and vertical computation (VeC). It has been shown that various loop structures in the cortical circuit play important roles in cortical computation as well as in memory storage and retrieval, keeping in conformity with the molecular basis of short and long term memory. A new learning scheme for the brain has also been proposed and how it is implemented within the proposed architecture has been explained. A few mathematical results about the architecture have been proposed, some of which are without proof.
| 19,003,474
|
Quantitative characterization of animal behavior following blast exposure.
|
The simplest approach to quantifying animal behavior begins by identifying a list of discrete behaviors and observing the animal's behavior at regular intervals for a specified period of time. The behavioral distribution (the fraction of observations corresponding to each behavior) is then determined. This is an incomplete characterization of behavior, and in some instances, mild injury is not reflected by statistically significant changes in the distribution even though a human observer can confidently and correctly assert that the animal is not behaving normally. In these circumstances, an examination of the sequential structure of the animal's behavior may, however, show significant alteration. This contribution describes procedures derived from symbolic dynamics for quantifying the sequential structure of animal behavior. Normalization procedures for complexity estimates are presented, and the limitations of complexity measures are discussed.
| 19,003,499
|
Dynamical aspects of behavior generation under constraints.
|
Dynamic adaptation is a key feature of brains helping to maintain the quality of their performance in the face of increasingly difficult constraints. How to achieve high-quality performance under demanding real-time conditions is an important question in the study of cognitive behaviors. Animals and humans are embedded in and constrained by their environments. Our goal is to improve the understanding of the dynamics of the interacting brain-environment system by studying human behaviors when completing constrained tasks and by modeling the observed behavior. In this article we present results of experiments with humans performing tasks on the computer under variable time and resource constraints. We compare various models of behavior generation in order to describe the observed human performance. Finally we speculate on mechanisms how chaotic neurodynamics can contribute to the generation of flexible human behaviors under constraints.
| 19,003,514
|
The neurodynamics underlying attentional control in set shifting tasks.
|
In this work we address key phenomena observed with classical set shifting tasks as the "Wisconsin Card Sorting Test" or the "Stroop" task: Different types of errors and increased response times reflecting decreased attention. A component of major importance in these tasks is referred to as the "attentional control" thought to be implemented by the prefrontal cortex which acts primarily by an amplification of task relevant information. This mode of operation is illustrated by a neurodynamical model developed for a new kind of set shifting experiment: The Wisconsin-Delayed-Match-to-Sample task combines uninstructed shifts as investigated in Wisconsin-like tasks with a Delayed-Match-to-Sample paradigm. These newly developed WDMS experiments in conjunction with the neurodynamical simulations are able to explain the reason for decreased attention in set shifting experiments as well the different consequences of decreased attention in tasks requiring bivalent yes/no responses compared to tasks requiring multivalent responses.
| 19,003,517
|
[Impact of motor and non-motor symptoms on the direct costs of Parkinson's disease].
|
We evaluate the health-related economic burden of patients with Parkinson's disease (PD) and identify the impact of motor and nonmotor symptoms (NMS) and health-related quality of life (HRQL) on the direct costs of PD in Spain. A cross-sectional, multicenter, pilot study covering consecutive PD outpatients who were included during the year 2004. Cognition (MMSE, SCOPA-Cog), autonomic dysfunction (SCOPA-AUT), motor impairment, severity and complications (SCOPA-motor), depression/anxiety (HADS), sleep (PDSS), psychosis (PPRS), pain, severity/global impairment (HY staging and CISI-PD) and health-related quality of life (EuroQoL) were evaluated. PD-related cost data were obtained using a standardized questionnaire. Association of clinical variables with direct costs was analyzed using the parametric and nonparametric tests. Seventy-eight consecutive PD outpatients (52% female and 48% male) having a mean age of 68.2 years (SD: 11), median HY stage of 2 (1-5), and mean PD duration of 8.3 years (SD: 5.4) were included in this study. Average 3 month total direct cost of PD was euro 2,631 (SD: 4507) per person (range: euro 71.5-euro 29,159) and medical treatment was the main cost driver, accounting for 34 % (mean drug cost per person euro 669; SD: 406). PD direct costs were significantly higher among younger patients, with higher HY stage, PD duration, severity, motor impairment and motor complications (p<0.05). Disease severity, motor impairment and complications had a higher impact on the direct costs of PD than NMS.
| 19,003,550
|
[Impaired fasting glucose as a risk factor for atherothrombotic or lacunar cerebral infarction. A case-control study].
|
Modifiable vascular risk factors (MVRF) for atherotrombotic or lacunar cerebral infarction have been clearly established, even though only a few population studies have been conducted and we do not know the risk of glycemic status prior to the diabetes mellitus (DM). This study has aimed to determine the relationship between impaired fasting glucose (IFG) and others MVRF with atherothrombotic or lacunar cerebral infarctions. A population based case-control study was conducted in two urban health care centers. 236 patients with atherothrombotic or lacunar CI, diagnosed by computed tomography scan magnetic resonance imaging. Unusual, indeterminate and cardioembolic cerebral infarction and cerebral hemorrhage were excluded. 441 patients without cerebral infarction, matched by age and gender, selected by means of simple random sampling. The odds ratios (OR) of IFG, DM, smoking, high blood pressure, hypercholesterolemia, hypertriglyceridemia, and obesity were measured with logistic regression. Mean age was 71.1 years old, 62 % being male. Prevalence in cases of MVRF was: IFG, 32.9%; DM, 49.6 %; smoking, 26.3%; high blood pressure, 78.8%; hypercholesterolemia, 29.3%; hypertriglyceridemia, 18.2%, and obesity, 63.5%. All MVRFs were associated in the crude analysis with increased risk of cerebral infarction. In the multivariate analysis, significant associations were found for IFG (OR: 3.2; 95 % confidence interval [CI]:1.1-7.9), DM (OR: 4.5; 95 % CI: 1.4-14.9), high blood pressure (OR: 2.6; 95 % CI: 1.5-4.5), hypercholesterolemia (OR: 2.3; 95 % CI: 1.4-3.9), and obesity (OR: 1.7; 95% CI: 1.0-2.9). Alterations of the glucose metabolism IFG and DM are important MVRF for atherothrombotic or lacunar cerebral infarction as seen by the high rate of OR and high prevalence in the population with cerebral infarction.
| 19,003,549
|
Improved synthesis of EM-1745, preparation of its C17-ketone analogue and comparison of their inhibitory potency on 17beta-hydroxysteroid dehydrogenase type 1.
|
Endocrine therapies are widely used for the treatment of estrogen-sensitive diseases. 17beta-hydroxysteroid dehydrogenase type 1 (17beta-HSD1) is involved in the last step of the biosynthesis of potent estrogen estradiol (E(2)). This enzyme catalyzes the reduction of the C17-ketosteroid estrone (E(1)) into the C17beta-hydroxy steroid E(2) using the cofactor NAD(P)H. The X-ray analysis of E(2)/adenosine bisubstrate inhibitor EM-1745 proven that this compound interacts with both the substrate- and the cofactor-binding sites. However, E(1) is a better substrate of 17beta-HSD1 than E(2). Thus, in order to improve the inhibitory potency of EM-1745, the C17-ketone analogue was prepared. During this work, a new and more efficient method for synthesizing EM-1745 was developed using an esterification and a cross-metathesis as key steps. Contrary to what was expected, the C17-ketone analogue of EM-1745 is a less potent inhibitor (IC(50) = 12 nM) than the C17-alcohol (IC(50) = 4 nM) in homogenated HEK-293 cells overexpressing 17beta-HSD1. Our results contribute to the knowledge of an unexpected observation: the C17-ketone steroidal inhibitors of 17beta-HSD1 are less potent than their corresponding C17-alcohol derivatives.
| 19,003,559
|
Substrate specificity of thymidine phosphorylase of E. coli: role of hydroxyl groups.
|
Substrate specificity of E. coli thymidine phosphorylase to pyrimidine nucleoside modified at 5'-, 3'-, and 2'-positions of sugar moiety has been studied. Equilibrium (K(eq)) and kinetics constants of phosphorolysis reaction of nucleosides were measured. The most important hydrogen bonds in enzyme-substrate complex have been determined.
| 19,003,566
|
Two new triterpenoid saponins from the flowers and buds of Lonicera japonica.
|
Two new triterpenoid saponins, loniceroside D (1) and loniceroside E (2), were isolated from the dry flowers and buds of Lonicera japonica Thunb., along with seven known compounds, chlorogenic acid (3), sweroside (4), vogeloside (5), epi-vogeloside (6), loniceroside A (7), loniceroside B (8), and loniceroside C (9). Their structures were elucidated on the basis of spectroscopic data, physicochemical properties, and acid hydrolysis.
| 19,003,609
|
Synthesis, structure, and bioevaluation of 2,5-bis(arylmethenyl)cyclopentanones.
|
Curcumin is an excellent lead compound with a variety of bioactivity. Recent articles reported that curcumin's instability and low bioavailability in vivo are mainly due to its easily decomposable beta-diketone moiety. With the aim of bioactive curcumin analogs with better pharmacokinetic property, we present here 11 bis(arylmethenyl)cyclopentanones similar to curcumin and without beta-diketone moiety by reacting relevant arylaldehydes and cyclopentanones. The analogs were structurally determined by 1HNMR and MS spectra, and the crystal structure of 6 was analyzed by X-ray diffraction. Their antibacterial activities in vitro against seven Gram-positive and Gram-negative bacteria were tested, and their inhibition of TNF-alpha and IL-6 secretion in LPS-induced mouse macrophages was investigated using enzyme-linked immunosorbent assay. It was observed that several derivatives displayed higher activity when compared with curcumin, and most of the analogs exhibited activities against the ampicillin-resistant Gram-negative Enterobacter cloacae.
| 19,003,615
|
Expression of the androgen receptor and syndecan-1 in breast tissue during different hormonal treatments in cynomolgus monkeys.
|
To analyze the expression of the androgen receptor(AR) and syndecan-1 in breast tissue during long-term hormonal treatment in cynomolgus monkeys. Sixty oophorectomized macaques were randomized to receive either tibolone, conjugated equine estrogens (CEE), CEE + medroxyprogesterone acetate (MPA) or no hormonal treatment. Breast tissue was collected at necropsy after 2 years and stained for AR and syndecan-1. Apparent differences were seen between treatment groups as compared to untreated animals. AR expression was markedly increased by tibolone and suppressed by combined CEE/MPA. Both treatments increased syndecan-1 in stromal tissue, whereas CEE alone had no significant effect. We found alternative regimens for hormonal therapy to differ in their influence on two markers of importance for the development of breast cancer. The results may be relevant for the ongoing clinical discussion on the long-term safety of different hormonal treatments.
| 19,003,632
|
Serum inhibin A level in preeclampsia and normotensive pregnancy.
|
To compare serum inhibin A levels in pregnancy complicated by preeclampsia and in normotensive pregnancy. Blood samples were taken from 60 women. Thirty women were diagnosed with preeclampsia, and 30 women had normotensive pregnancies. Both groups were matched for gestational age. Blood samples were collected in plain tubes, centrifuged, and stored at -80 degrees C until analyzed. All serum samples were measured for inhibin A level by enzyme-linked immunosorbent assay (ELISA). Inhibin A levels were greater in the preeclampsia group (1229.7 +/- 537.5 pg/mL) than in the normotensive group (839.1 +/- 370.0 pg/mL, p = 0.002). Levels of inhibin A in the preeclampsia group were greater than in the normotensive group.
| 19,003,635
|
Curable hypertension.
|
Hypertension due to a reninoma (suspected on the basis of biochemical and clinical features, and exquisite sensitivity of hypertension to angiotensin-converting enzyme [ACE] inhibition, but not visible on imaging) was cured by laparoscopic nephrectomy. Treatment was followed by an uneventful pregnancy.
| 19,003,641
|
A broader view of justice.
|
In this paper I argue that a narrow view of justice dominates the bioethics literature. I urge a broader view. As bioethicists, we often conceive of justice using a medical model. This model focuses attention at a particular point in time, namely, when someone who is already sick seeks access to scarce or expensive services. A medical model asks how we can fairly distribute those services. The broader view I endorse requires looking upstream, and asking how disease and suffering came about. In contrast to a medical model, a social model of justice considers how social determinants affect the health of a population. For example, social factors such as access to clean drinking water, education, safe workplaces, and police protection, profoundly affect risk for disease and early death. I examine one important social determinant of health, health care coverage, to show the limits of a medical model and the merits of a broader view.
| 19,003,692
|
Serum carotenoid concentrations in postmenopausal women from the United States with and without osteoporosis.
|
Antioxidant defenses may be compromised in osteoporotic women. Little is known about fruit and vegetable or carotenoid consumption among postmenopausal women. The primary carotenoids in human serum are alpha- and beta-carotene, lycopene, beta-cryptoxanthin, lutein, and zeaxanthin. This study investigated the interrelationships among serum carotenoid concentrations, fruit and vegetable intake, and osteoporosis in postmenopausal women (n = 59, 62.7 +/- 8.8 y). Bone density was assessed by dual energy x-ray absorptiometry and osteoporosis diagnosis was based upon T-scores. Serum samples (n = 53) and three-day diet records (n = 49) were analyzed. Logistic regression analyzed differences between carotenoids after adjusting for serum retinol; supplement usage; milk, yogurt, fruit, and vegetable intake; and body mass index (BMI). Pearson statistics correlated carotenoids with specific fruit or vegetable intake. Serum lycopene concentrations were lower in the osteoporosis group than controls (p = 0.03). Beta-cryptoxanthin intake was higher in the osteoporosis group (p = 0.0046). Total fruit and vegetable intakes were correlated with serum lycopene and beta-cryptoxanthin (p = 0.03, 0.006, respectively). Serum alpha-carotene concentration was associated with carrot intake, and zeaxanthin and beta-cryptoxanthin with lettuce intake. Carotenoids that may have beneficial skeletal effects are lower in women with osteoporosis. Research is needed to identify potential protective mechanisms or utilization of carotenoids during osteoporosis.
| 19,003,732
|
Effects of yogurt supplemented with fish oil on plasma lipid and glucose concentrations, and liver lipid contents in mice.
|
The aim of this study is to clarify the effects of yogurt supplemented with fish oil on plasma lipid and glucose concentrations, and hepatic lipid contents in mice. Male Crlj:CD-1 (ICR) mice were fed five experimental diets for 12 weeks. The experimental diets were as follows: without yogurt and fish oil (control diet); 10% (w/w) yogurt without fish oil [10% FO(-)]; 10% yogurt with fish oil [10% FO(+)]; 30% yogurt without fish oil [30% FO(-)]; 30% yogurt with fish oil [30% FO(+)]. Plasma triacylglycerol concentrations in the 10% FO(+) and 30% FO(-) groups were significantly lower than that in the control diet group (p < 0.05 and p < 0.05, respectively). Plasma total cholesterol and phospholipid concentrations were significantly lower in the 30% FO(+) group than in the control diet group (p < 0.005). Concentrations tended to be lower with supplementation with fish oil. Plasma glucose concentrations in the 10% FO(+) and 30% FO(+) groups were significantly lower than those in the control diet group (p < 0.005 and p < 0.01, respectively). Hepatic triacylglycerol and total cholesterol contents in the 30% FO(+) group were significantly lower than those in the control diet group (p < 0.05 and p < 0.005, respectively). These results suggest that plasma triacylglycerol and glucose concentrations are effectively decreased by supplementation of yogurt with fish oil.
| 19,003,735
|
Incidence of venous thrombosis following peripheral arterial interventions. A prospective study.
|
The objective of the study was to investigate the incidence of deep vein thrombosis (DVT) at the puncture site following peripheral interventions and to assess if there is a difference between using a vascular closure by means of vascular closure systems or compression bandages. We prospectively included 474 consecutive patients after peripheral arterial interventions. The day after peripheral arterial intervention we performed venous compression ultrasound to exclude DVT in the area of the groin. We recorded management of arterial closure and subsequent antithrombotic treatment of the patient. Four weeks after intervention follow-up was performed by phone to exclude clinical DVT, pulmonary embolism (PE), and death. We included 474 consecutive patients (mean age 69 y; 298 male / 176 female). All patients were under oral antiplatelet therapy. Vascular closure was achieved in 296 patients (62.44%) by Femostoptrade mark followed by compression bandage and in 178 (37.56 %) by using a vascular closure device alone. Sonography revealed no DVT the day after intervention, no clinical PE occurred. Four weeks follow-up showed no DVT, but there was one patient in the compression bandage group who had PE without proven deep vein thrombosis. Two patients died from other reasons than PE. The immediate and mid-term risk of DVT after peripheral arterial interventions is extremely low and is not increased if compression bandages are used for vascular closure.
| 19,003,747
|
[Analysis of nutritional status disorders in patients with chronic obstructive pulmonary disease].
|
Among the most common extrapulmonary manifestations of COPD are nutritional status disorders. The specific loss of weight, called cachexia, characterized by loss of lean body mass in some COPD patients is observed. The aim of the study was the quantitative and qualitative analysis of COPD patients' nutritional status disturbances. Fifty-five patients in different stages of COPD--43 males and 12 females (mean age 62.31 +/- 11.08) and 32 subjects from a control group (mean age 57.43 +/- 8.79) participated in the study. In both groups nutritional status was assessed using different indicators such as PIBW--percentage of ideal body weight, BMI--body mass index, FFMI--fat-free mass index and FMI--fat mass index. Malnutrition measured by PIBW, BMI, BMI percentiles, and FFMI was observed in 5.45%, 3.64%, 3.64% and 18.18% of COPD patients, respectively, and in the control group 3.12%, 0%, 3.12% and 3.12%, respectively. The BMI mean value did not differ significantly between groups. It was confirmed that cachexia assessed by FFMI occured more frequently in COPD patients than in the control group--19.05 kg/m2 vs. 20,55 kg/m2 (p = 0.04). 1. Nutritional status disorders pose a serious problem, which concerns about 1/5 of the COPD population. 2. It is necessary to perform quantitative analysis of nutritional status (assessment of lean and fat mass) because indicators of body mass (PIBW, BMI) are not sufficient for cachexia detection. 3. Having normal body mass does not exclude the possibility of nutritional status disorders in COPD patients.
| 19,003,762
|
Ephrin-A5 regulates the formation of the ascending midbrain dopaminergic pathways.
|
Dopaminergic neurons from the substantia nigra and the ventral tegmental area of the midbrain project to the caudate/putamen and nucleus accumbens, respectively, establishing the mesostriatal and the mesolimbic pathways. However, the mechanisms underlying the development of these pathways are not well understood. In the current study, the EphA5 receptor and its corresponding ligand, ephrin-A5, were shown to regulate dopaminergic axon outgrowth and influence the formation of the midbrain dopaminergic pathways. Using a strain of mutant mice in which the EphA5 cytoplasmic domain was replaced with beta-galactosidase, EphA5 protein expression was detected in both the ventral tegmental area and the substantia nigra of the midbrain. Ephrin-A5 was found in both the dorsolateral and the ventromedial regions of the striatum, suggesting a role in mediating dopaminergic axon-target interactions. In the presence of ephrin-A5, dopaminergic neurons extended longer neurites in in vitro coculture assays. Furthermore, in mice lacking ephrin-A5, retrograde tracing studies revealed that fewer neurons sent axons to the striatum. These observations indicate that the interactions between ephrin-A ligands and EphA receptors promote growth and targeting of the midbrain dopaminergic axons to the striatum.
| 19,003,794
|
Improved desorption electrospray ionization mass spectrometry performance using edge sampling and a rotational sample stage.
|
The position of the surface to be analyzed relative to the sampling orifice or capillary into the mass spectrometer has been known to dramatically affect the observed signal levels in desorption electrospray ionization mass spectrometry (DESI-MS). In analyses of sample spots on planar surfaces, DESI-MS signal intensities as much as five times greater were routinely observed when the bottom of the sampling capillary was appropriately positioned beneath the surface plane ('edge sampling') compared with when the capillary just touched the surface. To take advantage of the optimum 'edge sampling' geometry and to maximize the number of samples that could be analyzed in this configuration, a rotational sample stage was integrated into a typical DESI-MS setup. The rapid quantitative determination of caffeine in two diet sport drinks spiked with an isotopically labeled internal standard demonstrated the utility of this approach.
| 19,003,852
|
Omics-based profiling of carcinoma of the breast and matched regional lymph node metastasis.
|
Axillary lymph node (ALN) status is currently used as an important clinical indicator of breast cancer prognosis. However, the molecular mechanisms underlying lymph node metastasis are poorly understood and the relationship between ALN metastasis and the primary tumor remains unclear. In an effort to reveal structural changes in the genome and related protein responses that may drive regional metastatic progression we have analyzed matched pairs of primary breast tumors and ALN metastases both at the genomic and proteomic levels using comparative genomic hybridization (CGH) array, quantitative high-resolution 2-D PAGE in combination with MS, and immunohistochemistry (IHC). Array CGH revealed a remarkable similarity in genomic aberration profiles between the matched primary tumors and the ALN metastases. Quantitative profiling of 135 known proteins also revealed striking similarities in their overall expression patterns, although we observed distinct changes in the levels of individual proteins in some sample pairs. The remarkable similarities of the overall genomic and proteomic profiles between primary tumors and matched ALN metastases are taken to suggest that, in general, key biological characteristics of the primary breast tumor are maintained in the corresponding lymph node metastases. Given that the omics-based technologies are oblivious to changes that only occur in minor cellular subsets we validated the proteomic data using IHC, which provides protein expression information with a valuable topological component. Besides confirming the omics-derived data, the IHC analysis revealed that in two cases the ALN metastases may have been derived from a distinct minor cell subpopulation present in the primary tumor rather than from the bulk of it.
| 19,003,862
|
Identification of proteins that modify cataract of mouse eye lens.
|
The occurrence of a nuclear cataract in the eye lens due to disruption of the alpha3Cx46 connexin gene, Gja3, is dependent on strain background in a mouse model, implicating factors that modify the pathology. The differences upon cataractogenesis in the urea soluble proteins of the lens of two mouse strains, C57BL/6J and 129/SvJ, were analyzed by a comparative proteomics approach. Determination of the complete proteome of an organ offers the opportunity to characterize at a molecular level, differences in gene expression and PTMs occurring during pathology and between individuals. The abundance of 63 protein species was altered between the strains. A unique aspect of this study is the identification of chaperonin subunit 6A, mortalin, ERp29, and syntaxin-binding protein 6 in the eye lens. DNA polymorphisms resulting in nonconservative amino acid changes that led to altered physicochemical properties of the proteins were detected for mortalin, chaperonin subunit 6A, annexin A1, and possibly gamma-N crystallin. The results show HSP27/25 and/or ERp29 are the likely major modifying factors for cataractogenesis. Extension of the results suggests that small heat-shock proteins have a major role for influencing cataract formation in humans.
| 19,003,866
|
A method based on bead flows for spot detection on 2-D gel images.
|
A method has been developed to detect spots on 2-D gel images. It is based on an analogy with beads flowing uphill on the surface of the gel image and on the analysis of their paths. This method does not need image smoothing, thereby allowing for conservation of the resolution of the original image. In addition, it allows for the detection of overlapping spots, even when there is no valley between their centres.
| 19,003,871
|
Exhaled nitric oxide and respiratory symptoms in a community sample of school aged children.
|
To test the association between reported allergy and allergic diseases, respiratory symptoms, and the fractional concentration of exhaled nitric oxide (FeNO), in a community sample of school aged children. We administered a respiratory questionnaire and measured FeNO in a cross-sectional study of 1,135 children. FeNO was significantly greater in children with reported asthma (20.3 (standard deviation (SD) 21.3) parts per billion (ppb)) or allergies (18.1 (SD 18.0) ppb) than in healthy children (14.0 (SD 13.4) ppb). It was greater in children with asthma and reported allergies (22.8 (SD 23.6) ppb), than in children with asthma but no allergies (15.8 (SD 15.6) ppb) (overall P-value between disease groups = 0.002). FeNO was not related to respiratory symptoms in healthy children. Eczema was associated with an elevated FeNO concentration, even in the absence of respiratory symptoms. Some children with reported allergies but not asthma who had respiratory symptoms suggestive of asthma had elevated FeNO concentrations, and the proportion of healthy children with reported bronchitis or pneumonia in the past year who had an abnormally high FeNO concentration was significantly elevated. In a community sample of children, FeNO concentrations appear to reflect allergic conditions, including allergic asthma, reported allergies, and eczema, rather than just asthma, particularly since asthma in children may be non-allergic. FeNO is similarly elevated in school aged children with reported asthma or reported allergies. FeNO is higher in children with asthma and allergies than in children with asthma alone. However, an elevated FeNO may help alert the clinician to the possibility of undiagnosed asthma.
| 19,003,883
|
Neuroscientific approaches and applications within anthropology.
|
Many of the most distinctive attributes of our species are a product of our brains. To understand the function, development, variability, and evolution of the human brain, we must engage with the field of neuroscience. Neuroscientific methods can be used to investigate research topics that are of special interest to anthropologists, such as the neural bases of primate behavioral diversity, human brain evolution, and human brain development. Traditional neuroscience methods had to rely on investigation of postmortem brains, as well as invasive studies in living nonhuman primates. However, recent neuroimaging methods have made it possible to compare living human and nonhuman primate brains using noninvasive techniques such as structural and functional magnetic resonance imaging, positron emission tomography, and diffusion tensor imaging. These methods are providing an integrated picture of brain structure and function that was not previously available. With a combination of these traditional and modern neuroscience methods, we are beginning to explore and understand the neural bases of some of the most distinctive cognitive and behavioral attributes of the human species, including language, tool use, altruism, and mental self-projection, and we can now begin to propose plausible scenarios by which the neural substrates supporting these human specializations evolved from pre-existing neural circuitry serving related functions in common ancestors we shared with the living nonhuman primates. Consideration of the process of neurodevelopment suggests plausible mechanisms by which the highly encephalized human brain might have evolved. Neurodevelopmental studies also demonstrate that experience can shape both brain structure and function, providing a mechanism by which people of different cultures learn to act and think differently. Finally, not only can anthropologists benefit from neuroscience, neuroscience can benefit from the more sophisticated concept of evolution that anthropology offers, including an appreciation of evolutionary diversity as well as consideration of the process by which the human brain was formed during evolution.
| 19,003,891
|
Social organization of white-headed langurs (Trachypithecus leucocephalus) in the Nongguan Karst Hills, Guangxi, China.
|
The number of males per group is the most variable aspect of primate social organization and is often related to the monopolizability of females, which is mainly determined by the number of females per group and their reproductive synchrony. Colobines show both inter-specific and intra-specific variations in the number of males per group. Compared with other colobine species, little is known about the social organization of white-headed langur (Trachypithecus leucocephalus), despite its endangered status and unusual limestone habitat. As a part of a long-term study of the white-headed langurs in the Nongguan Karst Hills, Guangxi, China, we quantitatively investigated their social organization by analyzing census data from 1998 to 2003. The population censuses revealed that the predominant social organization of bisexual groups was the one-male group, similar to a previous report on this species and many other Asian colobines. In such groups, one adult male associated with 5.1 adult females, 0.1 sub-adult males, 2.6 juveniles and 2.9 infants on average, with a mean group size of 11.7 individuals. In addition, three multi-male groups were recorded, consisting of 2-3 adult males, 1-5 adult females, 0-2 sub-adult males, 0-7 juveniles and 0-2 infants. They did not contain more adult females than the one-male groups and were unstable in group membership. The langurs outside bisexual groups were organized into small nonreproductive groups or lived as solitaries. The nonreproductive groups averaged 1.3 adult males, 1.3 sub-adult males and 2.6 juveniles. Juvenile females were present in such groups on 52.4% of all occasions. As predicted by the monopolization model, the prevalence of the one-male pattern in this species may mainly be attributed to the small number of females in the group. The possible reasons for the occurrence of multi-male groups and the presence of juvenile females in nonreproductive groups are also discussed.
| 19,003,910
|
Understanding hind limb weight support in chimpanzees with implications for the evolution of primate locomotion.
|
Most quadrupedal mammals support a larger amount of body weight on their forelimbs compared with their hind limbs during locomotion, whereas most primates support more of their body weight on their hind limbs. Increased hind limb weight support is generally interpreted as an adaptation that reduces stress on primates' highly mobile forelimb joints. Thus, increased hind limb weight support was likely vital for the evolution of primate arboreality. Despite its evolutionary importance, the mechanism used by primates to achieve this important kinetic pattern remains unclear. Here, we examine weight support patterns in a sample of chimpanzees (Pan troglodytes) to test the hypothesis that limb position, combined with whole body center of mass position (COM), explains increased hind limb weight support in this taxon. Chimpanzees have a COM midway between their shoulders and hips and walk with a relatively protracted hind limb and a relatively vertical forelimb, averaged over a step. Thus, the limb kinematics of chimpanzees brings their feet closer to the COM than their hands, generating greater hind limb weight support. Comparative data suggest that these same factors likely explain weight support patterns for a broader sample of primates. It remains unclear whether primates use these limb kinematics to increase hind limb weight support, or whether they are byproducts of other gait characteristics. The latter hypothesis raises the intriguing possibility that primate weight support patterns actually evolved as byproducts of other traits, or spandrels, rather than as adaptations to increase forelimb mobility.
| 19,003,921
|
Structural elucidation of four new furostanol saponins from Tupistra chinensis by 1D and 2D NMR spectroscopy.
|
Four new furostanol saponins (1-4), two pairs of diastereoisomers, were isolated from methanolic extracts of Tupistra chinensis rhizomes and their structures were assigned from (1)H and (13)C NMR spectra, DEPT, and by 2D COSY, NOESY, HMQC, and HMBC experiments.
| 19,003,938
|
Immunological response to mistletoe (Viscum album L.) in cancer patients: a four-case series.
|
European mistletoe (Viscum album) has been used in complementary cancer treatment, but little is known concerning its effects on immunological parameters, although there is evidence that Viscum may stimulate the immune system. In this study, a trial was conducted with cancer patients to determine whether Viscum album extracts could improve the results of immune tests. These were: white blood cell count (leukocytes, neutrophils, lymphocytes), CD4+ and CD8+ T-lymphocytes, intradermal tests of delayed hypersensitivity (candidin, trichophytin, purified protein derivative-PPD), complement C3 and C4, and immunoglobulin A, G and M. Four patients received seven doses of subcutaneous Viscum album 20 mg, twice weekly. Immunological tests were carried out before and after treatment, and an increase in several parameters of humoral and cellular immunity were shown. Apart from reactions around the injection sites, treatment was well tolerated and all patients benefited from it. These results suggest that Viscum album can enhance humoral and cellular immune responses in cancer patients, but further studies attesting to the possible clinical impact of these immunological effects are necessary.
| 19,003,944
|
Mesenchymal stem cell secretion of chemokines during differentiation into osteoblasts, and their potential role in mediating interactions with breast cancer cells.
|
Over 70% of patients with advanced breast cancer will develop bone metastases for which there is no cure. Mesenchymal Stem Cells (MSCs) and their derivative osteoblasts are subpopulations of cells within the bone marrow environment, postulated as potential interacting targets for disseminating cancer cells because of their ability to secrete a range of chemokines. This study aimed to investigate chemokine secretion throughout MSC differentiation into osteoblasts and their effect on the breast cancer cells. Primary MSCs and osteoblast progenitors were cultured in appropriate conditions to induce differentiation into mature osteoblasts. Chemokines secreted throughout differentiation were detected using ChemiArray and ELISA. Migration of breast cancer cells in response to the bone-derived cells was quantified using Transwell inserts. Breast cancer cells were cocultured with MSCs, retrieved using magnetic beads, and changes in CCL2 expression were analyzed. MSCs secreted a range of factors including IL-6, TIMP-1 and CCL2, the range and level of which changed throughout differentiation. CCL2 secretion by MSCs increased significantly above control cells as they differentiated into mature osteoblasts (p<0.05). The bone-derived cells stimulated migration of breast cancer cells, and this was inhibited (21-50%) in the presence of a CCL2 antibody. CCL2 gene expression in breast cancer cells was upregulated following direct coculture with MSCs. The varying levels of chemokines secreted throughout MSC differentiation may play an important role in supporting tumor cell homing and progression. These results further highlight the distinct effect MSCs have on breast cancer cells and their potential importance in supporting development of metastases.
| 19,003,962
|
Rapid alterations in cortical protein profiles underlie spontaneous sleep and wake bouts.
|
Existing data indicate that sleep-wakefulness is an essential behavior. The biological function(s) of sleep, however, remains unknown, due, in part, to the lack of information available at the intracellular level. Preliminary microarray analyses show that changes in behavioral state influence regional mRNA profiles; however, the impact of sleep on protein signatures is virtually unexplored. In these studies, cortical protein profiles were examined after timed bouts of spontaneous sleep-wakefulness. Within minutes of each behavioral state examined, a small number of spots showing unique expression were detected. Mass spectroscopy analyses of sleep- and wake-related spots identified proteins associated with multiple functional categories. Two sleep-associated proteins were further validated using a sleep deprivation paradigm. We found preliminary evidence for two different post-transcriptional mechanisms-one (GAPDH) in which the amount of protein was increased in the recovery sleep following prolonged waking, while the other (actin) suggested that post-translational modifications may underlie sleep. The similarities between the effects of sleep on both protein and mRNA profiles indicate that dynamic intracellular changes underlie sleep-wake states and are consistent with roles for sleep in multiple biological functions.
| 19,003,977
|
A penalized likelihood approach for mixture cure models.
|
Cure models have been developed to analyze failure time data with a cured fraction. For such data, standard survival models are usually not appropriate because they do not account for the possibility of cure. Mixture cure models assume that the studied population is a mixture of susceptible individuals, who may experience the event of interest, and non-susceptible individuals that will never experience it. Important issues in mixture cure models are estimation of the baseline survival function for susceptibles and estimation of the variance of the regression parameters. The aim of this paper is to propose a penalized likelihood approach, which allows for flexible modeling of the hazard function for susceptible individuals using M-splines. This approach also permits direct computation of the variance of parameters using the inverse of the Hessian matrix. Properties and limitations of the proposed method are discussed and an illustration from a cancer study is presented.
| 19,003,981
|
Evaluation of molecular forms of prostate-specific antigen and human kallikrein 2 in predicting biochemical failure after radical prostatectomy.
|
Most pretreatment risk-assessment models to predict biochemical recurrence (BCR) after radical prostatectomy (RP) for prostate cancer rely on total prostate-specific antigen (PSA), clinical stage, and biopsy Gleason grade. We investigated whether free PSA (fPSA) and human glandular kallikrein-2 (hK2) would enhance the predictive accuracy of this standard model. Preoperative serum samples and complete clinical data were available for 1,356 patients who underwent RP for localized prostate cancer from 1993 to 2005. A case-control design was used, and conditional logistic regression models were used to evaluate the association between preoperative predictors and BCR after RP. We constructed multivariable models with fPSA and hK2 as additional preoperative predictors to the base model. Predictive accuracy was assessed with the area under the ROC curve (AUC). There were 146 BCR cases; the median follow up for patients without BCR was 3.2 years. Overall, 436 controls were matched to 146 BCR cases. The AUC of the base model was 0.786 in the entire cohort; adding fPSA and hK2 to this model enhanced the AUC to 0.798 (p=0.053), an effect largely driven by fPSA. In the subgroup of men with total PSA<or=10 ng/ml (48% of cases), adding fPSA and hK2 enhanced the AUC of the base model to a similar degree (from 0.720 to 0.726, p=0.2). fPSA is routinely measured during prostate cancer detection. We suggest that the role of fPSA in aiding preoperative prediction should be investigated in further cohorts.
| 19,003,994
|
Crystal structure of a 1.6-hexanediol bound tetrameric form of Escherichia coli lac-repressor refined to 2.1 A resolution.
|
We report the structure of a novel tetrameric form of the lactose repressor (LacI) protein from Escherichia coli refined to 2.1 A resolution. The tetramer is bound to 1.6-hexanediol present in the crystallization solution and the final R(free) for the structure is 0.201. The structure confirms previously reported structures on the monomer level. However, the tetramer is much more densely packed. This adds a new level of complexity to the interpretation of mutational effects and challenges details in the current model for LacI function. Several amino acids, previously associated with changes in function but unexplained at the structural level, appear in a new structural context in this tetramer which provides new implications for their function.
| 19,004,002
|
Genetic and epigenetic inactivation of LPL gene in human prostate cancer.
|
Lipoprotein lipase (LPL) is in chromosome 8p22, site of one of the most common somatic deletions in prostate tumors. Additionally, a CpG island (CGI) was identified in the LPL promoter region. To test the hypothesis that LPL is a tumor suppressor gene, which is inactivated by somatic deletion and hypermethylation in prostate cancer, we evaluated somatic DNA deletion and methylation status at LPL in 56 pairs of DNA samples isolated from prostate cancer tissues and matching normal controls and 11 prostate cell lines. We found that the DNA in 21 of 56 primary cancers (38%) was methylated in the LPL promoter CGI, whereas no methylation was detected in any normal samples. In addition, we found a hemizygous deletion at LPL in 38 of the 56 tumors (68%). When the results of deletion and methylation were considered together, we found LPL promoter CGI methylation occurred in 45% of LPL deleted tumors and in 22% of LPL retained tumors. Within several clinical characteristics tested, the preoperative PSA levels were found to be significantly higher in subjects with LPL promoter CGI methylation compared with subjects without LPL promoter methylation (p=0.0012). Additionally, demethylation of the LPL promoter CGI was accompanied by transcriptional reactivation of LPL in the prostate cancer cell lines DU145 and PC3. In summary, we report a novel finding that the LPL gene is commonly methylated in prostate tumors, and our results suggest that biallelic inactivation of LPL by chromosomal deletion and promoter hypermethylation may play a role in human prostate cancer.
| 19,004,026
|
Predicting patient dissatisfaction following joint replacement surgery.
|
The incidence of patient-reported dissatisfaction following total joint arthroplasty can be up to 30%. Our aim was to identify the preoperative patient-level predictors of patient dissatisfaction 1 year after surgery. We surveyed 1720 patients undergoing primary hip or knee replacement surgery. Relevant covariates including demographic data, body mass index, sex, comorbidities, and education were recorded. Joint functional status and patient quality of life were assessed at baseline and at 1-year followup with the Western Ontario McMaster University Osteoarthritis Index (WOMAC) and Medical Outcomes Study Short Form-36 (SF-36) scales, respectively. Patient satisfaction with surgery was determined with 4 survey questions at 1-year followup. There were no significant differences in demographic data between satisfied (n = 1290) and dissatisfied patients (n = 430). Logistic regression modeling showed that a lower preoperative SF-36 Mental Health score independently predicted patient dissatisfaction with surgery, adjusted for all relevant covariates (p < 0.05). We found no correlation between patient satisfaction and WOMAC change scores at 1-year followup (p = 0.31). Preoperative mental health is an important factor to consider when understanding patient satisfaction with surgery. Interventions to reduce psychological distress prior to surgery should be studied to determine if they may improve subjective outcomes of patients undergoing joint replacement surgery.
| 19,004,032
|
Serum autoantibodies and their clinical associations in patients with childhood- and adult-onset linear scleroderma. A single-center study.
|
To determine the frequency of selected serum autoantibodies and their clinical associations in patients with childhood-onset (ChO) or adult-onset (AO) linear scleroderma (LiScl) evaluated at a single institution. Seventy-two patients (ChO = 40, AO = 32), including 12 with en coup de sabre, were studied. All ChO patients had disease onset before age 16 years. Clinical features (extent of cutaneous disease, activity, and joint contractures) were recorded. Antinuclear antibodies (ANA) were identified by indirect immunofluorescence (HEp-2 cells), and anti-single-stranded DNA (anti-ssDNA), antihistone (AHA), and antichromatin (AChA) autoantibodies were detected by ELISA. There were no significant differences between groups in regard to gender, proportion with LiScl/E, or clinical features except joint contractures (ChO > AO; p = 0.04). There were no differences in the frequency of ANA or other autoantibodies between the groups except for AHA (ChO > AO). AHA was more frequently found with anti-ssDNA (p < 0.0001). LiScl patients with positive anti-ssDNA and/or AHA had more extensive cutaneous involvement and more often had joint contractures (p < 0.05). Anti-ssDNA was present more frequently in AO than in ChO patients with active lesions (p = 0.04). ANA and AChA were not associated with any clinical features. Both AHA and anti-ssDNA levels showed good correlation with disease severity. Over two-thirds of LiScl patients had ANA. Patients with ChO were similar to those with AO with regard to the frequency of selected serum autoantibodies. Anti-ssDNA and AHA were frequently found together and both were associated with more extensive skin disease with joint contractures. LiScl disease severity correlated with the serum levels of both these antibodies.
| 19,004,036
|
An investigation of ADA compliance of aquatic facilities in the North Texas area.
|
The study expands research on accessibility, comparing compliance scores of aquatic facilities in North Texas built before the 1991 Title III Americans with Disabilities Act Accessibility Guidelines (ADAAG) with facilities built after the 1991 ADAAG and the proposed 2002 supplement. A quasi-experimental design directed the selection of 52 facilities where measurements were taken to determine compliance with ADAAG and the supplement. A focus group provided insight into interpreting which features functioned as barriers or constraints to participation. Metropolitan statistical area in North Texas. A total of 52 aquatic facilities and 12 focus group participants (University of North Texas institutional review board 07-283). ADA aquatic facility compliance instrument. Frequency, ratios. No facilities were 100% ADA compliant overall, although some facilities were 100% compliant with specific structural domains. Women's restrooms rated lowest (average = 55%), and men's restrooms received the second lowest rating (average = 64%). Focus group results indicated that improperly designed restrooms and pool entries are primary barriers to participation. The findings support a need for stronger enforcement of policies that improve accessibility of facilities. Architectural reviews and construction practices need to be improved. The structural barriers and constraints identified can be limiting factors in efforts aimed at increasing physical activity among individuals with disabilities and individuals with physical limitations.
| 19,004,164
|
[Typical medical problems at the end of life: experiences in the intensive care unit].
|
Current regulations regarding living wills need to be revised in order to avoid conflicts between patients or their relatives/surrogates on one side, and physicians on the other. For wording the will the patient should seek not only legal, but also medical advice. For doctors treating the patient, it would be helpful if a written statement existed in which the patient identifies what kind of medical treatment he/she wants. This would apply to cases in which a patient's illness or injury precludes decision-making. Such a will could be stored on a still-to-be-established "health check card." By way of this card, the physician treating the patient could retrieve information about any living will in addition to the purely medical information. Despite potential gaps and uncertainties a patient's living will indicates to the physician that this patient has certain expectations regarding his/her treatment. In the event of a patient falling unconscious, the physician must talk to the relatives/surrogates in order to explore the patient's expectations about his/her treatment as nearly as closely as possible. This is the only way for the medical practitioner to meet his/her obligation towards the patient.
| 19,004,171
|
[Legal and practical aspects of euthanasia: experiences in The Netherlands].
|
Following a debate of more than 30 years the "Dutch Termination of Life on Request and Assisted Suicide Act" has come into force in 2002. Basically, the termination of life on request will in accordance with the Criminal Code remain punishable. Punishability is precluded if the physician notifies the review committee that he has performed termination of life on request and the committee is of the opinion that the physician has satisfied the due care criteria dictated in the Act. The Public Prosecution Service will not be informed, so the physician cannot be penalized. The review committee is composed of a lawyer, a physician and an expert on ethical issues.
| 19,004,180
|
Atherogenic lipoprotein subprofiling.
|
Elevated levels of low-density lipoprotein (LDL) cholesterol are a major risk factor for coronary heart disease. However, a significant number of coronary events occur in individuals with "normal" serum LDL cholesterol levels. It has been proposed that coronary heart disease (CHD) risk stratification may be improved by evaluating LDL particle size and number and lipoprotein (a) [Lp(a)] levels in addition to LDL-associated cholesterol levels. This chapter includes a brief review of lipoprotein biology and presents an overview of the association between novel lipoprotein markers and CHD risk. We also discuss methodologies currently available for atherogenic lipid subprofiling.
| 19,004,193
|
Biomarkers related to aging in human populations.
|
Biomarkers are increasingly employed in empirical studies of human populations to understand physiological processes that change with age, diseases whose onset appears linked to age, and the aging process itself. In this chapter, we describe some of the most commonly used biomarkers in population aging research, including their collection, associations with other markers, and relationships to health outcomes. We discuss biomarkers of the cardiovascular system, metabolic processes, inflammation, activity in the hypothalamic-pituitary axis (HPA) and sympathetic nervous system (SNS), and organ functioning (including kidney, lung, and heart). In addition, we note that markers of functioning of the central nervous system and genetic markers are now becoming part of population measurement. Where possible, we detail interrelationships between these markers by providing correlations between high risk levels of each marker from three population-based surveys: the National Health and Nutrition Examination Survey (NHANES) III, NHANES 1999-2002, and the MacArthur Study of Successful Aging. NHANES III is used instead of NHANES 1999-2002 when specific markers of interest are available only in NHANES III and when we examine the relationship of biomarkers to mortality which is only known for NHANES III. We also describe summary measures combining biomarkers across systems. Finally, we examine associations between individual markers and mortality and provide information about biomarkers of growing interest for future research in population aging and health.
| 19,004,190
|
Effects of the drug (BSZGC)-containing serum on proliferation of rat's osteoclasts and TRACP activity in vitro.
|
To observe effects of the drug-containing serum of Bu Shen Zhuang Gu Capsule (BSZGC Capsule for Tonifying the Kidney to Strengthen the Bones) on proliferation of the rat's osteoclasts and tartrate-resistant acid phosphatase (TRACP) activity in vitro so as to delve into the mechanisms of its preventive and therapeutic actions on osteoporosis. Forty female Sprague-Dawley rats aged three months were randomly divided into high dosage BSZGC group, medium dosage BSZGC group, low dosage BSZGC group, and the control group. BSZGC was orally administered into the rats of high, medium, and low dosage groups at different dosages for 12 days, and isometric normal saline was orally administered to the rats of the control group. The drug-containing serum and control serum were prepared. Osteoclasts isolated mechanically from the femur and tibia of Sprague-Dawley rats aged one week were cultured with medium added with different drug-containing sera and control serum. The growth of osteoclasts was observed under an inverted phase-contrast microscope, and optic density (OD) value of osteoclasts was determined by MTT colorimetric assay. TRACP activity was measured by the diazol method. OD value of osteoclasts in the high dosage drug-containing serum group, medium dosage drug-containing serum group, and low dosage drug-containing serum group was significantly lower than that in the control serum group (P < 0.05) with a dose-effect correlation. TRACP activity in high dosage drug-containing serum group, medium dosage drug-containing serum group, low dosage drug-containing serum group was significantly lower than that of the control serum group (P < 0.01), and no significant differences in TRACP activity were not found among the different dosages drug-containing serum groups. BSZGC can inhibit the proliferation of the osteoclasts and reduce TRACP activity, which may be one of the mechanisms of its preventive and therapeutic actions on osteoporosis.
| 19,004,206
|
[Haemorrhagic complications among patients on life-long acenocumarol therapy].
|
The aim of our study was to determine factors influencing occurrence of haemorrhagic events during acenocumarol therapy. A total of one hundred and forty patients were interviewed using a questionnaire. From one hundred forty (100%) of interviewed patients, forty one (29.3%) had haemorrhagic events and in eighteen (12.9%) cases the event followed the use of nonsteroid anti-inflammatory drugs (NSAID's) or paracetamol in combination with acenocumarol (AC). We found eleven cases of major bleeding in eight (5.7%) of patients Eight cases of bleeding were followed the use of NSAID's or paracetamol in combination with (AC). We found that there is a statistical relationship between the higher frequency of INR examination (at least as every four weeks) and decreased occurrence of haemorrhagic events (p<0.05). The frequency of INR examination should be done once every four weeks or more often, if there is evidence to suggest that testing more frequently. Monitoring of INR with frequency less than once in 4 weeks may increase the risk of bleeding in those patients. Concomitant use of NSAID's or paracetamol in combination with acenocumarol in patients without medical consultation can be associated with growing number of bleeding. Patients on AC therapy need to be inform about frequency of INR examination and acaenocumarol interactions with other medications, which are available without the prescription, to avoid haemorrhagic events.
| 19,004,232
|
Characterisation of actinomycetes isolated from ancient stone and their potential for deterioration.
|
Actinomycetes have been isolated from decayed and sound stone samples taken from a tomb site at Tell Basta, Zagazig City, Egypt. A total of 160 isolates have been characterised. The numbers and distribution of actinomycetes were studied during different seasons; during the winter months (18-20 degrees C), actinomycete numbers ranged from 10(3) to 10(4) cfu/g; in the summer (28-38 degrees C) lower counts were recorded. The actinomycete isolates were assigned to 4 different taxonomic groups: 54% belonged to the Streptomyces group, 26% to the Nocardia group, 14% showed the characteristics of the Micromonospora group, while the rest of the isolates analyzed (6%) were assigned to the sporangiate-type group of actinomycetes. The ability of the isolates to produce pigments as well as tolerance to high salinity were determined. It was shown that about 88% of the strains studied had the ability to produce extracellular pigments. Only 25% of the studied isolates showed tolerance to high salinity. The significance of actinomycetes to attack and degrade building stone was shown in laboratory experiments: actinomycetes recovered both from sound and decayed stones were capable of damaging stone under laboratory conditions as an up to 4% weight loss was recorded for some isolates.
| 19,004,242
|
Evaluation of antibacterial activity of synthetic aliphatic and aromatic monoacylglycerols.
|
The antibacterial activity of synthetic aliphatic and aromatic monoacylglycerols (MAGs) was studied against two human pathogens: Staphylococcus aureus and Escherichia coli. The active compounds inhibited selectively S. aureus. The most active compounds amongst them were those with medium size aliphatic chain and aromatic MAGs with electron withdrawing substituents at the aryl ring. The introduction of one or two-carbon spacer between the aryl ring and the carboxylic function did not influence antibacterial effectiveness.
| 19,004,249
|
[Selected applications of diode laser in laryngological surgery].
|
Diode laser due to emitted wavelength and achieved tissue effect appear to be universal tool for broad range of applications in laryngological surgery. The aim of the study was to present variety of applications of this type of laser in the treatment of selected pathologies of the nose, pharynx and oral cavity. 45 patients with various pathologies of the nose, pharynx and oral cavity were subjected to diode laser treatment under control of endoscopes and video routing system. In the studied group diode laser was used for hyperplastic turbinates treatment, correction of concha bullosa and septum deformations, ablation of nasal polyps and synechiae inside nasal cavity including cicatrical stenoses in the orifice of common lacrimal canalliculus, coagulation of vascular hemorrhagic diathesis, ablation of bleeding granulomas and polyps and treatment of vascular malformations. Results. Endoscopic diode laser surgery enabled for precise and save removal of the pathology accompanied only by minimal intraoperative bleeding. Healing and reepithelization process were normal and even in large postoperative defects do not exceeded 12 weeks. Postoperative complications in the form of cicatrix and synechiae were found in 3 patients. Various pathologies may be treated effectively with diode laser if appropriate laser beam parameters and surgical technique is applied.
| 19,004,255
|
[Reconstruction of the hypopharynx and cervical esophagus with gastric pull-up technique in the patient with recurrence carcinoma of the hypopharynx].
|
Patient with hypopharyngeal cancer are difficult to treat because they usually present with advanced disease, poor general health status and severe nutritional problems. Because of the poor prognosis, careful consideration has to be given regarding the choice of the correct surgical approach for respirato-digestive tract reconstruction. The authors present a case of recurrent hypopharyngeal cancer with cervical esophagus infiltration successfully treated with total laryngectomy and esophagectomy and gastric pull up reconstruction. Indications for technique, method of reconstruction and complications are, discussed. As most authors we consider the gastric transposition method as the preferred approach to restore digestive continuity after total esophagectomy. As the method is the single stage procedure it provides the best palliation of dysphagia and allows early resumption of an oral diet.
| 19,004,269
|
[CMV-induced cell death and fas gene expression in resting and proliferating human fibroblasts].
|
Cytomegalovirus (CMV) infection development and mRNA fas transcription levels (CD95) in resting (GO) and proliferating (S-phase) human lung embryo fibroblasts (HLEF-110044 line) were studied. In GO cells accumulation of infectious CMV was high and cell death was very quick, and fas gene expression was inhibited in early period of infection. In cells infected during S-phase CMV synthesis was lower and total cell death was detected only after 5 days; fas gene activity remained on high levels and increased during 6-48 hours. Death of CMV-infected fibroblasts occurred through apoptosis with cytopathic effect and detachment of cells in early stage, but without changes of cell membrane permeability and internucleosome fragmentation of DNA during later stages. In another HLEF-977 line CMV-induced apoptosis correlated with increased levels of fas gene transcription in resting cells. Positive association of activation Fas-receptor pathway and cell proliferation as well as different effect of CMV on activity of fas gene in 2 HLEF lines are discussed.
| 19,004,284
|
[Influence of antisense RNA and sequences of viral transactivators traps on RNA synthesis of HTLV-1 virus].
|
Significant number of scientific publications devoted to inhibition of viral replication by antisense RNA (asRNA) genes shows that this approach is useful for gene therapy of viral infections. To investigate the possibility of suppression of HTLV-1 virus reproduction by asRNA we constructed recombinant plasmids containing asRNA genes against U3 long terminal repeats region and X gene under the control of promoter of myeloproliferative sarcoma virus (MPSV) or without such promoter. Using stable calcium-phosphate transfection method with subsequent selection in the presence of G-418, RaHOS line-based cell clones carrying both asRNA genes and sequences able to bind HTLV-1 transactivator proteins (i.e. "traps" of viral transactivators, TVT) were obtained. Data from dot-hybridization analysis of viral RNA extracted from RaHOS cell clones showed that TVT sequences are able to suppress the viral RNA synthesis on 90% and asRNA against X gene synthesis--on 50%.
| 19,004,285
|
[Genetic identification as method of detection of pathogenic and symbiotic strains of enterococci].
|
Comparative study of 97 cultures of enterococci, including reference highly virulent strain Enterococcus faecalis ATCC 259212, 35 clinical isolates of E. faecium, 58 isolates of E. faecalis, and 3 probiotic strains of E. faecium: Linex (Lek company, Slovenia), SF68 (Bifiform, Denmark), and L3 (Avena, Russia). Thirteen of 93 cultures were isolated from patients treated with probiotic Linex, containing E. faecium (Linex) bacterium. Comparative analysis of cultures on the presence of 8 genes determining virulence (esp, asa1, efaA, cylA, cylM, gelE, sprE, and fsrB) was performed using polymerase chain reaction and testing with antibiotics. It was established that in some cases clinical cultures of enterococci used for analysis carried genes of pathogenicity and were not related to E. faecium (Linex) strain included in composition of the probiotic drug.
| 19,004,287
|
Unusual evolution of a residual dental cyst: a giant rhinolith.
|
Rhinoliths are calcareous concretions, consisting mainly of calcium and magnesium carbonates and phosphates. They can vary in size and shape, from small bodies to massive growths, and may invade adjacent structures. A 75-year-old female with a history of purulent discharge, nasal obstruction, and headache was observed in the ENT Department, University of Rome "Tor Vergata". Clinical examination revealed a giant rhinolith completely obstructing the right nasal cavity. Curiously, a dental CT scan showed a bony destruction area related to a residual dental cyst that had migrated superiorly into the maxilla. A transnasal, surgical approach was performed, and the rinolith was completely removed under local anaesthesia. The authors suggest that the giant rhinolith could have an odontogenic origin and that it could be an unusual complication of residual dental cyst.
| 19,004,311
|
[Comparison of effects of systemic (intramuscular) and intrastriatal injections of D1-dopamine receptor blocker on motor behavior and postural adjustment in dogs].
|
The study is based on the concept (Shapovalova, 2000) that activation and blockade of various types of muscarine and dopamine receptors located at various outputs of neostriatum should have different effects on motor behavior. The goal of the study was comparison of effects of systemic and intrastriatal injections of a selective blocker of D1 dopamine receptors on motor behavior. Experiments were carried out on 5 dogs on a model of instrumental defensive reflex (IDR) connected with keeping a certain flexor posture. Performance of the experiments, recording, storage, and analysis of data were accomplished with aid of original programs for PC. Systemic (intramuscular) administration of the blocker of D1 dopamine receptors SCH23390 at a dose of 0.025 mg/kg led to a sharp decrease of the IDR amplitude and in most cases--to the complete refusal of its performance. First of all, the phasic component of the instrumental response was inhibited, the diagonal pattern of the postural adjustment being preserved. Bilateral microinjections into the dog neostriatum of the same blocker of D1 receptors at a dose of 0.1 and 1.0 g did not change the percent of correct solution of the instrumental task, but at the same time produced a great number of changes in motorics: latent period of response increased statistically significantly, in some cases its amplitude rose, phasity decreased, intersignal raisings of limbs ceased completely. The obtained data allow the following conclusions to be made. 1. Difference of effects of systemic and intrastriatal injections of SCH23390 seems to be due to that at its systemic administration, D1 receptors of other structures, not only of neostriatum, can also be blocked. 2. Effect of nigrostriatal dopaminergic system on neostriatum via D1 receptors is complex - activation of motor activity (projectional spine neurons of the direct pathway) and a weak modulation of mental process (large spineless cholinergic interneurons). 3. Regulation of movement and postural adjustment is realized along different efferent neostriatum outputs.
| 19,004,318
|
[Molecular-genetic and epidemiologic examination of personnel subjected to occupational irradiation in the Russian Federal Nuclear Center (Sarov). Report 2. Genetic and non-genetic risk factors of morbidity].
|
Below is given a comparative evaluation of the genetic and the epidemiological examination of the cohort comprised by VNIIEF personnel subjected chronically to gamma-neutron ionizing radiation (74 persons), as well as nonirradiated cohort (74 persons). There are obtained data on the influence of the integral genotype structure on the health of an individual in case of occupational irradiation. The epidemiological analysis revealed statistically meaningful differences between the main and the control cohorts in the following five cases: acute respiratory diseases, diseases of upper respiratory tract, brain vessel malfunctions; illnesses connected with vision organs and cardio-vascular system and traumas. In such two cases as diseases of vision organs and cardio-vascular system the morbidity difference should be strictly attributed to the influence of the occupational factor. As a result of the genetic analysis it was shown that carriers of certain alleles of the loci ACP and Gc are credibly more susceptible to a number of diseases, when exposed to gamma-neutron irradiation. Besides, was proven that the occupational irradiation does not aggravate susceptibility to allergies and flew.
| 19,004,325
|
[Thiomorpholine transformation by the fungus Bjerkandera adusta].
|
A screening of lignin-degrading basidial fungi that can grow in the presence of thiomorpholine derivatives (the mixture of 1,4-perhydrothiazines) has been performed. Strain Bjerkandera adusta VKM F-3477 was shown to have the maximal rate of growth in the presence of these compounds, and its capacity for thiomorpholine degradation was studied. The methods of quantitative analysis of thiomorpholine and its degradation products on the basis of thin layer chromatography and high-performance liquid chromatography were developed. It was shown that the B. adusta strain did not utilize thiomorpholine as a carbon source but transformed it into thiomorpholine sulfoxide that accumulated in the medium. Mn peroxidase produced by B. adusta in the course of thiomorpholine transformation is not directly involved in its oxidation.
| 19,004,342
|
The right to access health care: health care according to European social security law instruments.
|
In this contribution we will look at how the traditional European social security conventions shape the fundamental right to health care. As to the instruments under investigation we focus upon the regional agreements that have been enacted within the framework of the Council of Europe. More specifically we will discuss how the (Revised) Social Charter and the minimum standard setting instrument (Code) give expression to the right to access to health care. This overview is then complemented by an analysis of recent case law of the European Court of Human Rights. The latter Court is indeed increasingly screening the national social security rules on their compatibility with the fundamental rights, as they are enshrined in the European Convention on Human Rights. Hence we will dwell upon the potential impact of this case law on the right to access health care. In the conclusions we will compare the three instruments regarding their legal interpretation of the right to health care.
| 19,004,379
|
Claiming access to health care in The Netherlands under international treaty law.
|
In the Netherlands, access to healthcare has been guaranteed by social health insurance legislation. But since the introduction of the health insurance system in the 1960s, it has been under frequent review, the latest reform was the introduction of regulated competition. At the same time, claiming health insurance entitlements under international (European) law has been changing, notably due to judicial activism.
| 19,004,383
|
Restaurant inspection frequency and food safety compliance.
|
Although food premises are regularly inspected, little information is available on the effect of inspections on compliance records, particularly with respect to the impact of the frequency of inspection on compliance. The following presents the outcome of a study designed to assess the impact of increased inspection frequency on compliance measures in Hamilton, Ontario, in the absence of any other changes to food handler/safety programs or legislation. High-risk food inspection premises were randomly assigned three, four, or five inspections per year. Results indicate that no statistical difference existed in outcome measures based on frequency of inspection. When premises were grouped based on the average time between inspections, premises with greater time between inspections scored better compliance measures relative to premises that were inspected more frequently. The study was also unique for the level of consultation and collaboration sought from the public health inspectors (PHIs) assigned to the Food Safety Program. Their knowledge and experience with respect to the critical variables associated with compliance were a complementary component to the literature review conducted by the research team.
| 19,004,396
|
A randomized prospective trial of the cutaneous and sensory effects of feminine hygiene wet wipes.
|
To assess the effects of feminine hygiene wet wipes on vulvar skin. An examiner-blind, randomized, prospective clinical trial of prototype wet wipes (treatment group) and dry toilet tissue (comparison group) in 120 premenopausal and 60 postmenopausal women was conducted in which the wipes or toilet tissues were for approximately 28 consecutive days. Use of wet wipes in lieu of toilet tissue caused no significant impact on erythema of the external genitalia. Mean vulvar erythema scores after 28 days of product use were lower than baseline scores for both wipes and toilet tissue. In postmenopausal women, wet wipes were associated with increased labial and perineal skin moisture. A minority of premenopausal participants found skin wetness from wipes less appealing and sometimes associated with slight sensory irritation; however, wet wipes were favored for personal cleansing by postmenopausal women. Four weeks of using wet wipes caused negligible clinical effects on vulvar skin irritation and appeared to contribute to skin moisture in postmenopausal women. This subgroup preferred using wet wipes, possibly due to the skin atrophy and dryness that accompany estrogen depletion. An opportunity exists to tailor the lotion level on the wipes m to the genital skin condition and preferences of younger and older Women.
| 19,004,402
|
Subgaleal hemorrhage after the use of a vacuum extractor during elective cesarean delivery: a case report.
|
Subgaleal hemorrhage (SGH) is a rare but possibly fatal condition in the neonate. Vacuum extractors are effective options for operative vaginal delivery. However, their use can increase the risk of intracranial hemorrhage in the neonate. A male infant delivered by cesarean section with assistance of a vacuum extractor had SGH attributed to the vacuum. The application of a vacuum extractor can increase the risk of trauma, including SGH. Good communication between health care providers and the use of the same precautions as with a vaginal delivery are advised.
| 19,004,410
|
MDMA-assisted psychotherapy using low doses in a small sample of women with chronic posttraumatic stress disorder.
|
The purpose of this study was to investigate the safety of different doses of MDMA-assisted psychotherapy administered in a psychotherapeutic setting to women with chronic PTSD secondary to a sexual assault, and also to obtain preliminary data regarding efficacy. Although this study was originally planned to include 29 subjects, political pressures led to the closing of the study before it could be finished, at which time only six subjects had been treated. Preliminary results from those six subjects are presented here. We found that low doses of MDMA (between 50 and 75 mg) were both psychologically and physiologically safe for all the subjects. Future studies in larger samples and using larger doses are needed in order to further clarify the safety and efficacy of MDMA in the clinical setting in subjects with PTSD.
| 19,004,414
|
Availability and characteristics of betel products in the U.S.
|
Betel use involves oral placement of shards of areca nut (Areca catechu palm seed containing the cholinergic agonist arecoline) wrapped with slaked lime in a betel leaf (Piper betle plant) or various chopped or powdered products containing areca nut and/or parts of the betel plant. Additives to this mixture include catechu (areca palm extract), spices/ sweeteners (e.g., saccharin, cloves), and/or tobacco. Betel use is most common in Asia and East India; however, little is known about the availability and characteristics of these products outside of this region. Thus, a representative sample of betel products and additives was purchased in the Richmond, Virginia area. Five venues were visited between March and May, 2006. Products successfully purchased were those containing betel alone (seven), betel/tobacco (three), tobacco alone (four), and additives (four). Most betel products listed ingredients on the packaging, though some did not explicitly distinguish between those with versus without tobacco. Importantly, seven of seven betel alone and one of three betel/tobacco products omitted any health-related warnings. All products were inexpensive and relatively obtainable in the groceries visited. More research is warranted in order to accurately estimate product emergence into the U.S. and other world markets, and the consequent impact on public health.
| 19,004,423
|
On-line flow injection-cloud point preconcentration of polycyclic aromatic hydrocarbons coupled with high-performance liquid chromatography.
|
Cloud point methodology has been used to develop a novel preconcentration and an analytical method for polycyclic aromatic hydrocarbons (PAHs) in soil sample. The nonionic surfactant Tergitol 15-S-7 was successfully used as the surfactant-mediated extractant in both ultrasonic and microwave-assisted extractions. Over 90% of recoveries for various PAHs in soil were obtained under the optimal experimental conditions. The extracts were then preconcentrated and analyzed by our proposed on-line coupling method "flow injection-cloud point preconcentration-high-performance liquid chromatography" (FI-CPP-HPLC) equipped with a fluorescence detector and an excitation/emission wavelength program. The preconcentration system and optimal working conditions were established. The limit of detection of the FI-CPE-HPLC system ranges from 0.101 to 0.456 microg/L for the selected PAHs, i.e., phenanthrene, pyrene, chrysene, benzo[k]fluoranthene and benzo[a]pyrene. Our proposed technique provides a reliable, simple and automatic analytical method for the determination of PAHs in environmental soil samples.
| 19,004,448
|
Geochemical characteristics and fluxes of organic carbon in a human-disturbed mountainous river (the Luodingjiang River) of the Zhujiang (Pearl River), China.
|
This study aims to investigate the state of the riverine organic carbon in the Luodingjiang River under human impacts, such as reforestation, construction of reservoirs and in-stream damming. Seasonal and spatial characteristics of total suspended sediment (TSS), dissolved organic carbon (DOC) and particulate organic carbon (POC), as well as C/N ratios and the stable carbon isotopic signatures of POC (delta(13)C(POC)) were examined based on a one-year study (2005) in the basin-wide scale. More frequent sampling was conducted in the outlet of the river basin at Guanliang hydrological station. DOC and POC concentrations showed flush effects with increasing water discharge and sediment load in the basin-wide scale. Atomic C/N ratio of POC had a positive relationship with TSS in the outlet of the basin, indicating the reduced aquatic sources and enhanced terrestrial sources during the high flood season. However, the similar relationship was not observed in the basin-wide scale mainly due to the spatial distributions of soil organic carbon and TSS. delta(13)C(POC) showed obvious seasonal variations with enriched values in the period with high TSS concentration, reflecting the increased contribution from C(4) plants with enhanced soil erosion. The specific flux of the total organic carbon (2.30 t km(-)(2) year(-1)) was smaller than the global average level. The ratio of DOC to POC was 1.17, which is higher than most rivers under Asian monsoon climate regime. The organic carbon flux was estimated to decline with decreasing sediment load as a result of reforestation, reservoir construction and in-stream damming, which demonstrates the impacts of human disturbances on the global carbon cycle.
| 19,004,473
|
Determination of phenolic endocrine disrupting chemicals and acidic pharmaceuticals in surface water of the Pearl Rivers in South China by gas chromatography-negative chemical ionization-mass spectrometry.
|
An analytical method for phenolic endocrine disrupting chemicals and acidic pharmaceuticals in river water was developed using gas chromatography mass spectrometry (GC-MS) coupled with negative chemical ionization (NCI) technique, and used for the determination of these compounds in the Pearl Rivers (Liuxi, Zhujiang and Shijing Rivers). Derivatization using pentafluorobenzoyl chloride (PFBOCl) and pentafluorobenzyl bromide (PFBBr) before GC-MS analysis were applied and optimized for phenolic compounds and acidic compounds, respectively. The target compounds were analyzed for river waters from the upstream to downstream of the Pearl Rivers. Phenolic compounds 4-tert-octylphenol (4-t-OP), 4-nonylphenol (4-NP), bisphenol-A (BPA), estrone (E1), estradiol (E2) and triclosan (TCS) were detected at trace or low levels in the water samples from Liuxi River and Zhujiang River. Diethylstilbestrol (DES) was not detected in the Pearl Rivers. The highest concentrations of the phenolic compounds were found in Shijing River, and they were 3150 ng/L for 4-t-OP, 11,300 ng/L for 4-NP, 1040 ng/L for BPA, 79 ng/L for E1, 7.7 ng/L for E2 and 355 ng/L for TCS, respectively. Only a few acidic pharmaceuticals were detected at low concentrations in water from Liuxi River and Zhujiang River, but the highest concentrations for the acidic pharmaceuticals were also found in Shijing River. The highest concentrations detected for clofibric acid, ibuprofen, gemfibrozil, naproxen, mefenamic acid and diclofenac were 17 ng/L, 685 ng/L, 19.8 ng/L, 125 ng/L, 24.6 ng/l and 150 ng/L, respectively. The results suggest Liuxi and Zhujiang Rivers are only slightly contaminated and can be used as drinking water sources, but Shijing River is heavily polluted by the wastewater from nearby towns.
| 19,004,474
|
Occurrence, properties and pollution potential of environmental minerals in acid mine drainage.
|
This paper describes the occurrences, the mineralogical assemblages and the environmental relevance of the AMD-precipitates from the abandoned mine of Valdarcas, Northern Portugal. At this mining site, these precipitates are particularly related with the chemical speciation of iron, which is in according to the abundance of mine wastes enriched in pyrrhotite and pyrite. The more relevant supergene mineralogical assemblages include the following environmental minerals: soluble metal-salts, mainly sulphates, revealing seasonal behaviour, iron-hydroxysulphates and iron-oxyhydroxides, both forming ochre precipitates of poorly and well-crystalline minerals. Pollution potential of the most highly water soluble salts was analysed in order to evaluate the environmental effect of their dissolution by rainfall. Laboratory experiments, carried out with iron and aluminium sulphates, demonstrated the facility to release metals, sulphate and acidity upon dissolution. Regarding the ochre precipitates, composed by several less soluble iron (III)-minerals, the spatial distribution on the nearby aqueous system as well as the proportion of Jarosite, Schwertmannite and Goethite in the mixtures gave information about the halo's contamination promoted by the AMD emerging from the waste-dumps.
| 19,004,477
|
Estrogen receptor modulators and estrogen receptor beta immunolabelling in human umbilical vein endothelial cells.
|
Human umbilical vein endothelial cells (HUVEC) exposed to the female sex hormone estradiol show different kinds of effects including increased elasticity, activation of plasma membrane Na+/H+ exchange, prostacyclin production, prevention of apoptosis and many others. The aim of this study was the systematic analysis of the immunolabelling of estrogen receptors (ERs), ERalpha and ERbeta, in HUVEC after stimulation with different commercially available ER modulators and ER agonists or antagonists. HUVEC response to these substances was shown to be regulated via ERbeta. ERalpha immunolabelling or up-regulation was abrogated after application of estrogen derivatives, selective estrogen receptor modulators (SERM) and ER agonists or antagonists. Immunolabelling of ERbeta was significantly increased by estradiol, estrone, ethinylestradiol and tumour necrosis factor alpha (TNFalpha). SERM, such as Tamoxifen, and pure antagonists, such as ICI 182.780, stimulated ERbeta in HUVEC at low concentrations, whereas higher concentrations inhibited ERbeta immunolabelling. The pure estrogen receptor agonist 2,3-bis (4-hydroxyphenyl) proprionitrile (DPN) exhibited its activating potential at low concentrations. In contrast, higher concentrations resulted in a down-regulation of ERbeta. Estrogenic effects in HUVEC, independent of stimulation or inhibition, are mediated via the ERbeta. SERM such as Tamoxifen and ER antagonists such as ICI 182.780 act as ER activators in low concentrations, whereas higher concentrations lead to inhibitory effects.
| 19,004,482
|
A new locus for congenital cataract, microcornea, microphthalmia, and atypical iris coloboma maps to chromosome 2.
|
To report a novel phenotype of autosomal dominant atypical congenital cataract associated with variable expression of microcornea, microphthalmia, and iris coloboma linked to chromosome 2. Molecular analysis of this phenotype may improve our understanding of anterior segment development. Observational case study, genome linkage analysis, and gene mutation screening. Three families, 1 Egyptian and 2 Belgians, with a total of 31 affected were studied. Twenty-one affected subjects and 9 first-degree relatives underwent complete ophthalmic examination. In the Egyptian family, exclusion of PAX6, CRYAA, and MAF genes was demonstrated by haplotype analysis using microsatellite markers on chromosomes 11, 16, and 21. Genome-wide linkage analysis was then performed using 385 microsatellite markers on this family. In the 2 Belgian families, the PAX6 gene was screened for mutations by direct sequencing of all exons. Phenotype description, genome-wide linkage of the phenotype, linkage to the PAX6, CRYAA, and MAF genes, and mutation detection in the PAX6 gene. Affected members of the 3 families had bilateral congenital cataracts inherited in an autosomal dominant pattern. A novel form of hexagonal nuclear cataract with cortical riders was expressed. Among affected subjects with available data, 95% had microcornea, 39% had microphthalmia, and 38% had iris coloboma. Seventy-five percent of the colobomata were atypical, showing a nasal superior location in 56%. A positive lod score of 4.86 was obtained at theta = 0 for D2S2309 on chromosome 2, a 4.9-Mb common haplotype flanked by D2S2309 and D2S2358 was obtained in the Egyptian family, and linkage to the PAX6, CRYAA, or MAF gene was excluded. In the 2 Belgian families, sequencing of the junctions and all coding exons of PAX6 did not reveal any molecular change. We describe a novel phenotype that includes the combination of a novel form of congenital hexagonal cataract, with variably expressed microcornea, microphthalmia, and atypical iris coloboma, not caused by PAX6 and mapping to chromosome 2. The authors have no proprietary or commercial interest in any materials discussed in this article.
| 19,004,499
|
Self-medication of mood disorders with alcohol and drugs in the National Epidemiologic Survey on Alcohol and Related Conditions.
|
Using alcohol or drugs to reduce emotional distress (self-medication) has been proposed as an explanation for the high comorbidity rates between anxiety and substance use disorders. Self-medication has been minimally studied in mood disorders despite equally high rates of alcohol and drug use. Data came from the National Epidemiologic Survey on Alcohol and Related Conditions (NESARC), a large (n=43,093, age 18 years and older) nationally representative survey of mental illness in community-dwelling adults. Prevalence rates of self-medication were determined for DSM-IV mood disorders: dysthymia, major depressive disorder, bipolar I disorder, and bipolar II disorder. Multiple logistic regression generated odds ratios for the association between each category of self-medication and anxiety and personality disorders. Almost one-quarter of individuals with mood disorders (24.1%) used alcohol or drugs to relieve symptoms. The highest prevalence of self-medication was seen in bipolar I disorder (41.0%). Men were more than twice as likely as women to engage in self-medication (Adjusted Odds Ratio=2.18; 95% Confidence Interval 1.90-2.49). After controlling for the effects of substance use disorders, self-medication was associated with higher odds of comorbid anxiety and personality disorders when compared to individuals who did not self-medicate. Cross-sectional design. The use of alcohol and drugs to relieve affective symptoms is common among individuals with mood disorders in the general population, yet is associated with substantial psychiatric comorbidity. These findings may help clinicians identify a subgroup of people with mood disorders who suffer from a higher mental illness burden.
| 19,004,504
|
Subsets and Splits
No community queries yet
The top public SQL queries from the community will appear here once available.