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In re Rodas.

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Brain Diseases↗

Control of cell shape and elongation by the rodA gene in Bacillus subtilis.

The Escherichia coli rodA and ftsW genes and the spoVE gene of Bacillus subtilis encode membrane proteins that control peptidoglycan synthesis during cellular elongation, division and sporulation respectively. While rodA and ftsW are essential genes in E. coli, the B. subtilis spoVE gene is dispensable for growth and is only required for the synthesis of the spore cortex peptidoglycan. In this work, we report on the characterization of a B. subtilis gene, designated rodA, encoding a homologue of E. coli RodA. We found that the growth of a B. subtilis strain carrying a fusion of rodA to the IPTG-inducible Pspac promoter is inducer dependent. Limiting concentrations of inducer caused the formation of spherical cells, which eventually lysed. An increase in the level of IPTG induced a sphere-to-short rod transition that re-established viability. Higher levels of inducer restored normal cell length. Staining of the septal or polar cap peptidoglycan by a fluorescent lectin was unaffected during growth of the mutant under restrictive conditions. Our results suggest that rodA functions in maintaining the rod shape of the cell and that this function is essential for viability. In addition, RodA has an irreplaceable role in the extension of the lateral walls of the cell. Electron microscopy observations support these conclusions. The ultrastructural analysis further suggests that the growth arrest that accompanies loss of the rod shape is caused by the cell's inability to construct a division septum capable of spanning the enlarged cell. RodA is similar over its entire length to members of a large protein family (SEDS, for shape, elongation, division and sporulation). Members of the SEDS family are probably present in all eubacteria that synthesize peptidoglycan as part of their cell envelope.

Artificial Gene Fusion↗

The UDCA dosage deficit: a fate shared with CDCA.

Ursodeoxycholic acid (UDCA) is used both as the treatment of choice in many cholestatic syndromes and as complementary therapy in many liver diseases. However, few dose-finding studies exist, and none has evaluated the efficacy and long-term safety of UDCA therapy in primary biliary cirrhosis (PBC). There is an open debate about UDCA's impact on the natural history of PBC, and no universal evidence of benefits on the major endpoint exists. This is perhaps due to a UDCA dosage deficit. Most clinical trials on PBC therapy have used conservative dosages of UDCA similar to those of chenodeoxycholic acid (CDCA) used for dissolution of gallstones. It may be necessary to re-evaluate the dosage of UDCA that provides the most effective treatment.

Chenodeoxycholic Acid↗

Chemiluminescent immunoperoxidase assay for the dot blot hybridization detection of Parvovirus B19 DNA using a low light imaging device.

A new synthesized stable trifluoro-substituted acridan (2',3',6'-trifluorophenyl 10-methylacridan-9-carboxylate known as Lumigen PS-3) has been applied as chemiluminescent substrate of the horseradish peroxidase (HRP) enzyme (neutral isoenzyme C) in a dot blot hybridization assay for the detection of B19 Parvovirus DNA. The dot blot hybridization assay uses digoxigenin-labeled DNA probes which are immunoenzymatically revealed by anti-digoxigenin Fab fragments conjugated with HRP. The results obtained using PS-3 reagent or the luminol-based enhanced chemiluminescence detection system (ECL Amersham and Renaissance DuPont NEN kits) were compared. A high-performance, low-intensity-light imaging luminograph apparatus to collect light emission was used. The detection systems using the different chemiluminescent substrates gave sensitive and reproducible results for calibration graphs, with high precision (relative standard deviation 5-18%). With the chemiluminescent assay it was possible to detect 0.5, 1, or 2 pg of target homologous DNA, using PS-3, ECL, or Renaissance (RE) reagents, respectively, while colorimetry had a detection limit of 5 pg. When clinical samples were analyzed the positive reference sera and the PCR-positive products gave light emissions with values higher than background at 2 sigma level, while the negative samples gave a signal comparable to the background noise for all chemiluminescent reagents. The PS-3 reagent detected one more dilution (1/256) than ECL and RE (1/128) of positive reference sera.

DNA, Viral↗

Heritability of running economy: a study made on twin brothers.

Running economy (RE), defined as the steady-state of oxygen uptake (VO2) for a given running velocity, is a factor of sports performance the genetic component of which has seldom been reported to date. We studied this component using a heritability index (HI) in a group of 32 male twins, 8 monozygotic (MZ) and 8 dizygotic (DZ) pairs, all sportsmen with similar perinatal and environmental backgrounds. Zygocity was determined by the identity of erythrocytic antigenic, protein and enzymatic polymorphism, and human leucocyte antigen serologic types between co-twins. The subjects exercised twice on a treadmill, once until exhaustion and again at submaximal intensities. Pulmonary gas exchange was measured continuously using an automatic analyser system during both tests. Blood samples were obtained during the recovery period to determine lactate concentrations. No significant differences were observed between MZ and DZ, in respect of RE at any speed or in maximal VO2 relative to body mass. Nevertheless, significant HI (P < 0.05) was found in maximal lactate concentrations (HI=0.75) and in respiratory equivalent for oxygen at two speeds, 7 km x h(-1) HI=0.71) and 8 km x h(-1) (HI=0.79), differences which probably suggest that there are differences in RE. In conclusion, we did not detect a genetic component in RE or in maximal oxygen uptake, but a genetic component for markers of anaerobic metabolism was present.

Adult↗

Galantamine and memantine produce different degrees of neuroprotection in rat hippocampal slices subjected to oxygen-glucose deprivation.

Recent clinical trials have shown that galantamine is efficacious in the treatment of mild to moderate Alzheimer's and vascular dementia, and memantine in severe stages of these diseases. Hence, the hypothesis that these two drugs might exert different degrees of neuroprotection has been tested. Rat hippocampal slices were subjected to oxygen and glucose deprivation (OGD) and to a re-oxygenation period. Neuronal damage was monitored using the lactate dehydrogenase (LDH) released into the Krebs-bicarbonate medium as an indicator. Galantamine, a mild acetylcholinesterase (AChE) blocker and nicotinic receptor modulator, given 30 min before and during OGD plus re-oxygenation (1, 2 and 3 h) significantly reduced LDH release by around 50%. Galantamine 5 microM reduced LDH release significantly during the re-oxygenation period while at 15 microM it afforded significant reduction of LDH release both during OGD and re-oxygenation. Memantine, a reversible blocker of NMDA receptors, at 10 microM only significantly reduced (40%) LDH release after 3 h re-oxygenation. The classical NMDA blocker MK-801 reduced LDH released around 40% at 1 microM at all re-oxygenation times studied. These data indicate that galantamine has a neuroprotective window against anoxia wider than memantine. Whether these differences can be clinically relevant remain to be studied in appropriate clinical trials.

Animals↗

How large is the optic disc? Systematic errors in fundus cameras and topographers.

PURPOSE: To determine whether or not there are systematic differences in the areas of optic discs as measured by different machines using different measurement algorithms and whether racial or gender differences exist in optic disc area measurements. METHODS: We examined the results of twenty-three published studies on the size of normal optic discs of various patient populations. Studies differed in the type of instrument and method used to measure optic disc area, and the number, age, race and gender of subjects examined. Noticing that different machines exhibited statistically significant systematic differences in optic disc sizes of comparable populations, we computed a "normalization" factor for each machine based on these mean differences. Applying this normalization factor to the results, we then re-examined the differences between racial and gender groups. RESULTS: By comparing the results of mean optic disc areas of different racial groups made with different machines, and normalizing results according to those of the Zeiss fundus camera, we found the normalization factors for the following machines to be, Zeiss fundus camera: 1 (by definition), Rodenstock Optic Disc Analyzer (RODA): 1.51, Topcon fundus camera: 1.04, Heidelberg Retina Tomograph (HRT): 1.15 and TopSS scanning laser ophthalmoscope: 1.29. That is, to bring the results of area measurements made with a RODA machine in line with those made with a Zeiss fundus camera, one should multiply the former by the factor 1.51. Using the normalized results, we confirmed the findings of previous authors that the optic disc areas of black subjects were statistically significantly larger than those of white subjects (n-weighted mean effect = 0.556 +/- 0.142 S.E., n = 5). Further, the meta-analysis of various racial populations from five studies shows that males have significantly larger discs than females (n-weighted mean effect = 0.151 +/- .055 S.E., n = 9). CONCLUSION: Different machines and techniques give different results when populations of similar racial composition are measured. We recommend applying the above normalizing factors when comparing studies that employ different instruments.

Adolescent↗

Demonstration of a tool for automatic learning and re-use of knowledge in the activated sludge process.

Wastewater treatment plant operators encounter complex operational problems related to the activated sludge process and usually respond to these by applying their own intuition and by taking advantage of what they have learnt from past experiences of similar problems. However, previous process experiences are not easy to integrate in numerical control, and new tools must be developed to enable re-use of plant operating experience. The aim of this paper is to investigate the usefulness of a case-based reasoning (CBR) approach to apply learning and re-use of knowledge gained during past incidents to confront actual complex problems through the IWA/COST Benchmark protocol. A case study shows that the proposed CBR system achieves a significant improvement of the benchmark plant performance when facing a high-flow event disturbance.

Artificial Intelligence↗