Helicobacter pylori VacA cytotoxin interacts with fibronectin and alters HeLa cell adhesion and cytoskeletal organization in vitro

Helicobacter pylori vacuolating cytotoxin VacA causes multiple effects on epithelial cell function and morphology, but the effects of VacA on signal transduction pathways and the cytoskeleton have not been investigated in detail. In this study, we analyzed the effects of native VacA on HeLa and AGS cell adhesion to fibronectin and laminin under serum-free conditions. Confocal microscopic examination revealed increased number of cells with rounded morphology and inhibition of actin fiber formation, in the presence of VacA. VacA binds to fibronectin in vitro in a dose-dependent manner. This interaction was partly inhibited by a peptide containing an arginine-glycine-aspartic acid motif. The adhesion of HeLa cells to fibronectin, but not to laminin, was decreased in the presence of VacA. Thus, VacA may interact with fibronectin and influence integrin receptor-induced cell signaling and cytoskeleton-dependent cell functions.     

A Study of Glutathione <Emphasis Type="Italic">S</Emphasis>-transferase pi Expression in Central Nervous System of Subjects with Amyotrophic Lateral Sclerosis Using RNA Extraction from Formalin-Fixed,Paraffin-Embedded Material

The expression of glutathione S-transferase pi (GST pi), an enzyme responsible for inactivation of a large variety of toxic compounds was studied in spinal cord, motor and sensory brain cortex obtained from patients who died in the course of amyotrophic lateral sclerosis (ALS). The studies were performed on formalin-fixed, paraffin-embedded (FFPE) and freshly frozen tissues. The method of RNA isolation from FFPE was modified. A significant decrease of GST pi-mRNA expression was found in cervical spinal cord and motor brain cortex of ALS subjects comparing to analogue control tissues (P < 0.01), as well as in motor cortex of ALS subjects comparing to their sensory cortex (P < 0.05). In spinal cords the decrease in GST pi-mRNA expression was accompanied by a decrease of GST pi protein level. Results indicated lowered GST pi expression on both mRNA and protein levels in the regions of nervous system affected by ALS. The non-properly inactivated by GST toxic electrophiles and organic peroxides may thus contribute to motor neurons damage.     

Age effect of broiler breeders on fertility and sperm penetration of the perivitelline layer of the ovum

The objective of this study was to determine the age effect of a broiler breeder flock on duration of fertility and number of spermatozoa penetrating the perivitelline layer overlying the germinal disc (SP/mm(2) GDIPVL). Moreover, in the second half of the flock's reproductive life, the effect of using ejaculates of young roosters (CA2) in artificial insemination (AI) on the above parameters of fertility was estimated. The commercial flock of broiler breeder hens (n = 100) was inseminated six times from 31 to 62 weeks of age. Additional inseminations, with ejaculates of roosters aged 31 and 36 weeks (CA2), were performed at 56 and 62 weeks of age. AI was performed during two consecutive days (D0 and D1) with an insemination dose of 125 x 10(6) spermatozoa/0.06 ml containing pooled ejaculates. The following parameters were studied: the effective and maximum duration of fertility (De and Dm), percent of fertility on different days after AI (FD10, FD15 and FD20), indices of duration of sperm penetration (DSP, SP < or = 3/GDIPVL), SP/mm(2) GDIPVL in eggs laid on successive days after insemination of hens at different age, and correlations between some fertility indices. Both for De and Dm, the highest values were noted after AI of the layers at 36 weeks of age (14.8 +/- 0.49 and 17.4 +/- 0.46 days, respectively), which were about 2 days longer than at 56 weeks. All fertility indices decreased gradually with age, starting from AI at peak egg production (31-36 weeks of age), while the use of ejaculates from CA2 did not help to increase them significantly. Correlation coefficients between SP/mm(2) GDIPVL and the other fertility indices were positive and highest for eggs laid on D3. It is concluded that high De values can be obtained from broiler breeders in adequate environmental and technological conditions of AI. It is suggested that the age-related decrease in fertility is more pronounced in females, in which the efficiency of sperm storage tubules decreases. The present fertility indices indicate the possibility of lengthening AI intervals, especially at peak egg production.     

Metabolic memory - the implications for diabetic complications

Many important biochemical mechanisms are activated in the presence of high levels of glucose, which occur in diabetes. Large randomised studies have established that early intensive glycaemic control reduces the risk of diabetic complications. This phenomenon has recently been dubbed 'metabolic memory'. It has been suggested that early glycaemia normalisation can halt the hyperglycaemia-induced pathological processes associated with enhanced oxidative stress and glycation of cellular proteins and lipids. The phenomenon of metabolic memory suggests that early aggressive treatment and strict glycaemic control could prevent chronic diabetic complications.     

Thar She Blows! A Novel Method for DNA Collection from Cetacean Blow

Background

Molecular tools are now widely used to address crucial management and conservation questions. To date, dart biopsying has been the most commonly used method for collecting genetic data from cetaceans; however, this method has some drawbacks. Dart biopsying is considered inappropriate for young animals and has recently come under scrutiny from ethical boards, conservationists, and the general public. Thus, identifying alternative genetic collection techniques for cetaceans remains a priority, especially for internationally protected species.

Methodology/Principal Findings

In this study, we investigated whether blow-sampling, which involves collecting exhalations from the blowholes of cetaceans, could be developed as a new less invasive method for DNA collection. Our current methodology was developed using six bottlenose dolphins, Tursiops truncatus, housed at the National Aquarium, Baltimore (USA), from which we were able to collect both blow and blood samples. For all six individuals, we found that their mitochondrial and microsatellite DNA profile taken from blow, matched their corresponding mitochondrial and microsatellite DNA profile collected from blood. This indicates that blow-sampling is a viable alternative method for DNA collection.

Conclusion/Significance

In this study, we show that blow-sampling provides a viable and less invasive method for collection of genetic data, even for small cetaceans. In contrast to dart biopsying, the advantage of this method is that it capitalizes on the natural breathing behaviour of dolphins and can be applied to even very young dolphins. Both biopsy and blow-sampling require close proximity of the boat, but blow-sampling can be achieved when dolphins voluntarily bow-ride and involves no harmful contact.     

Influence of Methionine upon the Activity of Antioxidative Enzymes in The Kidney of Rats Exposed to Sodium Fluoride

The intensified or uncontrolled formation of reactive oxygen species leads to disturbances of numerous biochemical processes. Among the factors inducing intensified free radical processes, fluoride ions are listed, among others. One of the organs most exposed to the toxic activity of fluorides is the kidney. In the study presented here, the influence of fluorine upon the activity of selected antioxidant enzymes in rat kidney has been examined, as well as antioxidant properties of methionine during intoxication with sodium fluoride. The experiment was carried out on Wistar FL rats (adult females) that for 35 days were administered water, NaF, NaF with methionine (doses: 10 mg NaF/kg bw/day, 10 mg Met/kg bw/day) . The influence of administered NaF and Met upon the antioxidative system in kidney was examined by analyzing the activity of the most important antioxidative enzymes (SOD, CAT, GPX, GR, GST). The studies carried out confirmed the disadvantageous effect of NaF upon the antioxidative system in rats (decrease in activity of antioxidative enzymes). Methionine increased the activity of antioxidative enzymes, most efficiently that of GPX, GR, and GST.     

Antiproliferative activity of salinomycin and its derivatives

Antiproliferative activity of seven amides and one benzotriazole ester derivative of salinomycin, a polyether ionophore antibiotic, with recently reported antibacterial activity, are herein described. Salinomycin and the majority of derivatives exhibit potent antiproliferative activity against the drug-resistant cancer cell lines. Moreover almost all derivatives show stronger activity against LoVo/DX cell line than that of unmodified salinomycin.     

Organization of Patient Management and Fungal Epidemiology in Cystic Fibrosis

The achievement of a better life for cystic fibrosis (CF) patients is mainly caused by a better management and infection control over the last three decades. Herein, we want to summarize the cornerstones for an effective management of CF patients and to give an overview of the knowledge about the fungal epidemiology in this clinical context in Europe. Data from a retrospective analysis encompassing 66,616 samples from 3235 CF patients followed-up in 9 CF centers from different European countries are shown.