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1.
The galactose operon encoding a repressor and genes for the Leloir pathway for galactose metabolism (galactokinase, galactose-1-phosphate-uridyl transferase and UDP glucose-4-epimerase) was located adjacent to the multiple sugar metabolism (msm) operon on the chromosome of Streptococcus mutans Ingbritt (serotype c) and the complete nucleotide sequence of this 5-kilobase region was determined. The Leloir pathway was induced by the presence of galactose in the growth medium or following the release of intracellular galactose after uptake and cleavage of -galactosides by the multiple sugar metabolism system. Analysis of the mechanism of galactose transport confirmed the absence of a galactose-specific phosphotransferase system and suggested the presence of an inducible galactose permease. Evidence is presented that galactose transport is independent of the proton motive force and may be ATP-dependent. 相似文献
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M. Ristanovic V. Bunjevacki C. Tulic I. Novakovic A. Nikolic 《Russian Journal of Genetics》2007,43(6):705-708
The aim of this study was to detect frequency of microdeletions of Y chromosome in idiopathic cases of male infertility in
Serbian population. Patients were subjected to detailed clinical, endocrinological and cytogenetic examinations. Ninety patients
with normal cytogenetic findings with azoospermia and severe oligozoospermia were included in the study. In these patients
microdeletion analysis was performed by multiplex polymerase chain reaction (PCR) method on DNA extracted from peripheral
blood. In each case 6 markers in azoospermia factor (AZF) regions were tested: sY84, sY86 (AZFa); sY127, sY134 (AZFb); sY254,
sY255 (AZFc). Deletions on the Y chromosome were detected in 14 of 90 cases (15.6%), nine with azoospermia and five with severe
oligozoospermia. Of total number of 17 deletions, 11 (64.7%) were detected in AZFc region, three (17.6%) in AZFa region and
three (17.6%) in AZFb region. Microdeletions in AZF region of the Y chromosome, especially AZFc microdeletions, represent
common genetic cause of idiopathic azoospermia and severe oligozoospremia in Serbian infertile men. Therefore, testing for
Y chromosome microdeletions should be considered as an important element in diagnosis and genetic counseling of infertile
men in Serbia and decisions regarding the assisted reproduction should be made based on the presence and type of AZF microdeletions.
The text was submitted by the authors in English. 相似文献
4.
Aleksandra Topic Marina Milenkovic Snezana Uskokovic-Markovic Dragana Vucicevic 《Biological trace element research》2010,134(3):296-306
Investigations of effective, orally active, and safe antidiabetic metallopharmaceuticals have been carried out during the
last two decades. It has been reported that tungsten compounds mimic the action of insulin in intact cell systems. As insulin
mimetics, the most investigated tungsten compound was sodium tungstate (ST), rarely investigated was tungstophosphoric acid
(WPA), but never alanine complex of tungstophosphoric acid (WPA-A). In this study, the insulin mimetic activity of three different
tungsten compounds, ST, WPA, and WPA-A, was evaluated by means of in vitro measurements of the glucose uptake and inhibition
of free fatty acids release from epinephrine-treated isolated rat white adipocytes. We investigated the influence of concentration
(lower and higher, 0.1 and 1.0 mM, respectively) and solvent: isotonic salt solution—saline (0.9% w/v of NaCl) and dimethyl sulfoxide (DMSO; 2% v/v), on the biological effect of tested compounds. Our experimental data showed that all of the three investigated tungsten
compounds possess insulin mimetic activity in vitro on the isolated adipocytes. Influence of concentration and solvents on
insulin mimetic effect for the certain tungsten compounds were: WPA was shown effect independently of concentration and solvents;
higher concentration and DMSO were significant decreasing insulin mimetic effect of ST; lower concentration and saline led
to decreasing effect of WPA-A. Generally, there were no differences in insulin mimetic effect of three tungsten compounds
in lower concentration and dissolved in DMSO. When saline was used as solvent, it was needed higher concentration of investigated
compounds to accomplish the same effect. In conclusion, our results suggest that low concentration (0.1 mM) of ST, WPA, and
WPA-A dissolved in 2% DMSO could be the good candidates for in vivo investigation of their antidiabetic properties. 相似文献
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Timothy D. Wiltshire Dragana Milosevic Eapen K. Jacob Stefan K. Grebe Allan B. Dietz 《Cytotherapy》2021,23(5):452-458
Background AimsViral vectors are commonly used to introduce chimeric antigen receptor (CAR) constructs into cell therapy products for the treatment of human disease. They are efficient at gene delivery and integrate into the host genome for subsequent replication but also carry risks if replication-competent lentivirus (RCL) remains in the final product. An optimal CAR T-cell product should contain sufficient integrated viral material and no RCL. Current product testing methods include cell-based assays with slow turnaround times and rapid quantitative polymerase chain reaction (PCR)-based assays that suffer from high result variability. The authors describe the development of a droplet digital PCR (ddPCR) method for detection of the vesicular stomatitis virus G glycoprotein envelope sequence, required for viral assembly, and the replication response element to measure integration of the CAR construct.MethodsAssay validation included precision, linearity, sensitivity, specificity and reproducibility over a range of low to high concentrations.ResultsThe limit of detection was 10 copies/μL, whereas negative samples showed <1.3 copies/μL. Within and between assay imprecision coefficients of variation across the reportable range (10–10 000 copies/μL) were <25%. Accuracy and linearity were verified by comparing known copy numbers with measured copy numbers (R2 >0.9985, slope ~0.9). Finally, serial measurements demonstrated very good long-term reproducibility (>95% of replicate results within the originally established ± two standard deviations).ConclusionsDDPCR has excellent reproducibility, linearity, specificity and sensitivity for detecting RCL and assuring the safety of patient products in a rapid manner. The technique can also likely be adapted for the rapid detection of other targets during cell product manufacturing, including purity, potency and sterility assays. 相似文献
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Miloš R. Filipović Dragana Stanić Smiljana Raičević Mihajlo Spasić 《Free radical research》2013,47(1):62-72
The present study demonstrates that manganese superoxide dismutase (MnSOD) (Escherichia coli), binds nitric oxide (√NO) and stimulates its decay under both anaerobic and aerobic conditions. The results indicate that previously observed MnSOD-catalyzed √NO disproportionation (dismutation) into nitrosonium (NO+) and nitroxyl (NO? ) species under anaerobic conditions is also operative in the presence of molecular oxygen. Upon sustained aerobic exposure to √NO, MnSOD-derived NO? species initiate the formation of peroxynitrite (ONOO? ) leading to enzyme tyrosine nitration, oxidation and (partial) inactivation. The results suggest that both ONOO? decomposition and ONOO? -dependent tyrosine residue nitration and oxidation are enhanced by metal centre-mediated catalysis. We show that the generation of ONOO? is accompanied by the formation of substantial amounts of H2O2. MnSOD is a critical mitochondrial antioxidant enzyme, which has been found to undergo tyrosine nitration and inactivation in various pathologies associated with the overproduction of √NO. The results of the present study can account for the molecular specificity of MnSOD nitration in vivo. The interaction of √NO with MnSOD may represent a novel mechanism by which MnSOD protects the cell from deleterious effects associated with overproduction of √NO. 相似文献
8.
Dragana Dobrijevic Gaetana Di Liberto Kosei Tanaka Tomas de Wouters Rozenn Dervyn Samira Boudebbouze Johan Binesse Hervé M. Blottière Alexandre Jamet Emmanuelle Maguin Maarten van de Guchte 《PloS one》2013,8(6)
Complex microbial ecosystems are increasingly studied through the use of metagenomics approaches. Overwhelming amounts of DNA sequence data are generated to describe the ecosystems, and allow to search for correlations between gene occurrence and clinical (e.g. in studies of the gut microbiota), physico-chemical (e.g. in studies of soil or water environments), or other parameters. Observed correlations can then be used to formulate hypotheses concerning microbial gene functions in relation to the ecosystem studied. In this context, functional metagenomics studies aim to validate these hypotheses and to explore the mechanisms involved. One possible approach is to PCR amplify or chemically synthesize genes of interest and to express them in a suitable host in order to study their function. For bacterial genes, Escherichia coli is often used as the expression host but, depending on the origin and nature of the genes of interest and the test system used to evaluate their putative function, other expression systems may be preferable. In this study, we developed a system to evaluate the role of secreted and surface-exposed proteins from Gram-positive bacteria in the human gut microbiota in immune modulation. We chose to use a Gram-positive host bacterium, Bacillus subtilis, and modified it to provide an expression background that behaves neutral in a cell-based immune modulation assay, in vitro. We also adapted an E. coli – B. subtilis shuttle expression vector for use with the Gateway high-throughput cloning system. Finally, we demonstrate the functionality of this host-vector system through the cloning and expression of a flagellin-coding sequence, and show that the expression-clone elicits an inflammatory response in a human intestinal epithelial cell line. The expression host can easily be adapted to assure neutrality in other assay systems, allowing the use of the presented presentation system in functional metagenomics of the gut and other ecosystems. 相似文献
9.
Natasa M. Milic Goran Z. Trajkovic Zoran M. Bukumiric Andja Cirkovic Ivan M. Nikolic Jelena S. Milin Nikola V. Milic Marko D. Savic Aleksandar M. Corac Jelena M. Marinkovic Dejana M. Stanisavljevic 《PloS one》2016,11(2)
Background
Although recent studies report on the benefits of blended learning in improving medical student education, there is still no empirical evidence on the relative effectiveness of blended over traditional learning approaches in medical statistics. We implemented blended along with on-site (i.e. face-to-face) learning to further assess the potential value of web-based learning in medical statistics.Methods
This was a prospective study conducted with third year medical undergraduate students attending the Faculty of Medicine, University of Belgrade, who passed (440 of 545) the final exam of the obligatory introductory statistics course during 2013–14. Student statistics achievements were stratified based on the two methods of education delivery: blended learning and on-site learning. Blended learning included a combination of face-to-face and distance learning methodologies integrated into a single course.Results
Mean exam scores for the blended learning student group were higher than for the on-site student group for both final statistics score (89.36±6.60 vs. 86.06±8.48; p = 0.001) and knowledge test score (7.88±1.30 vs. 7.51±1.36; p = 0.023) with a medium effect size. There were no differences in sex or study duration between the groups. Current grade point average (GPA) was higher in the blended group. In a multivariable regression model, current GPA and knowledge test scores were associated with the final statistics score after adjusting for study duration and learning modality (p<0.001).Conclusion
This study provides empirical evidence to support educator decisions to implement different learning environments for teaching medical statistics to undergraduate medical students. Blended and on-site training formats led to similar knowledge acquisition; however, students with higher GPA preferred the technology assisted learning format. Implementation of blended learning approaches can be considered an attractive, cost-effective, and efficient alternative to traditional classroom training in medical statistics. 相似文献10.
Biochar,Bentonite and Zeolite Supplemented Feeding of Layer Chickens Alters Intestinal Microbiota and Reduces Campylobacter Load 总被引:1,自引:0,他引:1
Tanka P. Prasai Kerry B. Walsh Surya P. Bhattarai David J. Midmore Thi T. H. Van Robert J. Moore Dragana Stanley 《PloS one》2016,11(4)
A range of feed supplements, including antibiotics, have been commonly used in poultry production to improve health and productivity. Alternative methods are needed to suppress pathogen loads and maintain productivity. As an alternative to antibiotics use, we investigated the ability of biochar, bentonite and zeolite as separate 4% feed additives, to selectively remove pathogens without reducing microbial richness and diversity in the gut. Neither biochar, bentonite nor zeolite made any significant alterations to the overall richness and diversity of intestinal bacterial community. However, reduction of some bacterial species, including some potential pathogens was detected. The microbiota of bentonite fed animals were lacking all members of the order Campylobacterales. Specifically, the following operational taxonomic units (OTUs) were absent: an OTU 100% identical to Campylobacter jejuni; an OTU 99% identical to Helicobacter pullorum; multiple Gallibacterium anatis (>97%) related OTUs; Bacteroides dorei (99%) and Clostridium aldenense (95%) related OTUs. Biochar and zeolite treatments had similar but milder effects compared to bentonite. Zeolite amended feed was also associated with significant reduction in the phylum Proteobacteria. All three additives showed potential for the control of major poultry zoonotic pathogens. 相似文献