Pathogenicity of Aeromonas hydrophila artificially infected to Indian major carp (Cirrhinus mrigala Ham.)

Abrasion, feeding, injection and immersion methods were used to evaluate the pathogenicity of five different strains of Aeromonas hydrophila viz. RG (rui gill), ML (mrigal lesion), SG (sharpunti gill), F1K (mrigal kidney), GFL (gold fish lesion) and Ah-19 (Aeromonas hydrophila-19, Ref. Strain) against C. mrigala H. Bacterial suspension containing viable cells of 7.5x 10⁵ per ml was found to be very effective in intramuscular injection and feeding resulting 100% mortality after 96hr of inoculation. The strain RG, ML and F1K produced scale loss with erosion of the skin surface with/without hemorrhagic lesion after 48hr of inoculation following abrasion method. The strains SG and Ah-19 resulting scale loss with erosion of the skin surface with/without hemorrhagic lesion after 72hr of inoculation following abrasion and injection methods. SG and F1K caused reddening in mouth region after 72hr of feeding inoculation, whereas RG resulted frank ulcers from eroded dermal layer exposing underlying musculature which was hemorrhagic after 96hr of inoculation by abrasion method.

Bacterial flora associated with diseased fish and their antibiogram

Bacterial flora associated with tail rot/fin rot of Carassius auratus, Xiphophorus helleri and hemorrhagic ulcers of Clarias spp were studied. Sensitivity pattern of 33 isolates comprising Aeromonas spp, Pseudomonas spp and Gram-positive rods from diseased C. auratus, X. helleri and Clarias spp were screened against six broad-spectrum antibiotics viz. ciprofloxacin, chloramphenicol, co-trimoxazole, gentamycin, nitro-furantoin and oxytetracycline. Ciprofloxacin was the most effective in inhibiting bacteria at 0.05-0.10 µg/ml level. About 44% of Pseudomonas spp. was resistant to nitrofurantoin. Resistance to oxytetracycline was seen in 27% of Aeromonas spp Gram-positive rods were comparatively more resistant to antibiotics. The multiple antibiotic resistances were seen in 21% of the bacterial isolates of diseased fish.

Gastroprotective potential of Buddleja scordioides Kunth: Effects into the modulation of antioxidant enzymes and inflammation markers in an in vivo model

Ethnopharmacological relevance: A common plant used to treat several gastric disorders is Buddleja scordioides Kunth,commonly known as salvilla. Aim of thes tudy: To detect inflammatory markers,in order to evaluate the gastroprotective potential of salvilla infusions,as this could have beneficial impact on the population exposed to gastric ulcers and colitis. Materials and methods: The present work attempted infusions were prepared with B. scordioides (1% w/w) lyophilized and stored.Total phenolic content and GC–MS analysis were performed. Wistar rats were divided into five groups a negative vehicle control,an indomethacin group,and three experimental groups,named preventive,curative,and suppressive. All rats were sacrificed under deep ether anesthesia(6h)after the last oral administration of indomethacin/infusion.The rat stomachs were promptly excised,weighed,and chilled in ice-cold and 0.9%NaCl.Histological analysis,nitrites quantification and immunodetection assays were done. Results: B.scordioides infusions markedly reduced the visible hemorrhagic lesions induced byindomethacin in rat stomachs,also showed down-regulation of COX2, IL-8 and TNFα and up-regulation of COX-1with a moderate down-regulation of NFkB and lower amount of nitrites.However,this behavior was dependent on the treatment,showing most down-regulation of COX-2,TNFα and IL-8 in the curative treatment;more down-regulation of NF-kB in the preventive treatment;and more up-regulation of COX-1 for the suppressor and preventive treatments. Conclusion: The anti-inflammatory potential of B. scordioides infusions could be related with the presence of polyphenols as quercetin in the infusion and how this one is consumed.

Helicobacter pylori: comparative genomics and structure-function analysis of the flagellum biogenesis protein HP0958

Helicobacter pylori is a gastric pathogen which infects ~50% of the global population and can lead to the development of gastritis, gastric and duodenal ulcers and carcinoma. Genome sequencing of H. pylori revealed high levels of genetic variability; this pathogen is known for its adaptability due to mechanisms including phase variation, recombination and horizontal gene transfer. Motility is essential for efficient colonisation by H. pylori. The flagellum is a complex nanomachine which has been studied in detail in E. coli and Salmonella. In H. pylori, key differences have been identified in the regulation of flagellum biogenesis, warranting further investigation. In this study, the genomes of two H. pylori strains (CCUG 17874 and P79) were sequenced and published as draft genome sequences. Comparative studies identified the potential role of restriction modification systems and the comB locus in transformation efficiency differences between these strains. Core genome analysis of 43 H. pylori strains including 17874 and P79 defined a more refined core genome for the species than previously published. Comparative analysis of the genome sequences of strains isolated from individuals suffering from H. pylori related diseases resulted in the identification of “disease-specific” genes. Structure-function analysis of the essential motility protein HP0958 was performed to elucidate its role during flagellum assembly in H. pylori. The previously reported HP0958-FliH interaction could not be substantiated in this study and appears to be a false positive. Site-directed mutagenesis confirmed that the coiled-coil domain of HP0958 is involved in the interaction with RpoN (74-284), while the Zn-finger domain is required for direct interaction with the full length flaA mRNA transcript. Complementation of a non-motile hp0958-null derivative strain of P79 with site-directed mutant alleles of hp0958 resulted in cells producing flagellar-type extrusions from non-polar positions. Thus, HP0958 may have a novel function in spatial localisation of flagella in H. pylori

Helicobacter pylori modulation of gastric and duodenal mucosal T cell cytokine secretions in children compared with adults.

BACKGROUND: In contrast to adults, ulcers are un-common in Helicobacter pylori-infected children. Since immunological determinants influence the outcome of H. pylori infection, we have investigated mucosal T cell responses in H. pylori-infected children and compared them with those of adults and negative controls. MATERIAL AND METHODS: Mucosal biopsies were obtained from 43 patients undergoing an upper GI endoscopy for dyspeptic symptoms. The concentrations of released cytokines and the density of CD3+, CD25+ and CD69+cells were evaluated by flow cytometry, and the numbers of cytokine-secreting cells were measured by ELISPOT. RESULTS: The numbers of isolated antral CD3+ lymphocytes were only significantly raised in infected adults compared with noninfected controls (p < 0.05), whereas the proportion of CD3+ cells expressing activation markers (CD25 or CD69) remained low. In the stomach, IFN-gamma concentrations increased in infected children and infected adults compared with controls (p < 0.05), but IFN-gamma concentrations were tenfold lower in children than in adults (p < 0.01). IL-2, IL-4, IL-10 and TNF-alpha concentrations were similar in infected and in uninfected children and adults. In contrast, in the duodenum, IFN-gamma, as well as IL-4 and IL-10 concentrations were only increased in infected children compared with controls (p < 0.05). The concentrations of these cytokines were similar in both groups of adults who, however, like children, displayed a higher number of duodenal IL-4-secreting cells compared to controls (p < 0.05). CONCLUSION: These results suggest that IFN-gamma secretion in the stomach of H. pylori-infected patients is lower in children than in adults. This could protect children from development of severe gastro-duodenal diseases such as ulcer disease. In addition, infected patients are characterised by a dysregulation of the mucosal cytokine secretion at distance from the infection site.

The Application of an Exogenous Linear and Radial Electrical Field to an In Vitro Chronic Diabetic Ulcer Model for Evaluation as a Potential Treatment

Chronic diabetic ulcers affect approximately 15% of patients with diabetes worldwide. Currently, applied electric fields are being investigated as a reliable and cost-effective treatment. This in vitro study aimed to determine the effects of a constant and spatially variable electric field on three factors: endothelial cell migration, proliferation, and angiogenic gene expression. Results for a constant electric field of 0.01 V demonstrated that migration at short time points increased 20-fold and proliferation at long time points increased by a factor of 1.40. Results for a spatially variable electric field did not increase directional migration, but increased proliferation by a factor of 1.39 and by a factor of 1.55 after application of 1.00 V and 0.01 V, respectively. Both constant and spatially variable applied fields increased angiogenic gene expression. Future research that explores a narrower range of intensity levels may more clearly identify the optimal design specifications of a spatially variable electric field.

Expression, purification and X-ray crystallographic analysis of the Helicobacter pylori blood group antigen-binding adhesin BabA

Helicobacter pylori is a human pathogen that colonizes about 50% of the world's population, causing chronic gastritis, duodenal ulcers and even gastric cancer. A steady emergence of multiple antibiotic resistant strains poses an important public health threat and there is an urgent requirement for alternative therapeutics. The blood group antigen-binding adhesin BabA mediates the intimate attachment to the host mucosa and forms a major candidate for novel vaccine and drug development. Here, the recombinant expression and crystallization of a soluble BabA truncation (BabA^25-460) corresponding to the predicted extracellular adhesin domain of the protein are reported. X-ray diffraction data for nanobody-stabilized BabA^25-460 were collected to 2.25Å resolution from a crystal that belonged to space group P21, with unit-cell parameters a = 50.96, b = 131.41, c = 123.40Å, α = 90.0, β = 94.8, γ = 90.0°, and which was predicted to contain two BabA^25-460-nanobody complexes per asymmetric unit.

Los diferentes tipos de láser y sus aplicaciones en podología

Los diferentes tipos de láseres, sobre todo el láser de diodo, irrumpen en la terapéutica podológica para proporcionar una alternativa más de tratamiento en muchas patologías que son el día a día de las consultas. El buen manejo y el conocimiento de sus características son requisitos imprescindibles para no tener efectos secundarios indeseados y poder llevar a cabo tratamientos poco dolorosos, minimizando el tiempo total, y muchas veces proporcionando una solución a diversas patologías.

Fermentation of heated gluten systems by gut microflora

The Maillard reaction causes changes to protein structure and occurs in foods mainly during thermal treatment. Melanoidins, the final products of the Maillard reaction, may enter the gastrointestinal tract, which is populated by different species of bacteria. In this study, melanoidins were prepared from gluten and glucose. Their effect on the growth of faecal bacteria was determined in culture with genotype and phenotype probes to identify the different species involved. Analysis of peptic and tryptic digests showed that low molecular mass products are formed from the degradation of melanoidins. Results showed a change in the growth of bacteria. This in vitro study demonstrated that melanoidins, prepared from gluten and glucose, affect the growth of the gut microflora.

MODULATORY ACTION OF HELICOBACTER-PYLORI ON HISTAMINE-RELEASE FROM MAST-CELLS AND BASOPHILS IN-VITRO

Helicobacter pylori is important in the aetiology of peptic ulceration. Despite inducing an inflammatory response in the mucosa, the organism persists, suggesting that it has efficient protective mechanisms. Some bacterial and viral products modulate histamine secretion from inflammatory cells. Therefore, this study examined the modulatory effects of H. pylori preparations on histamine release from rat peritoneal mast cells and human basophils. Eleven clinical isolates of H. pylori were prepared in different ways: as whole washed bacteria, washed sonicated bacteria, and formalin-killed bacteria, and as outer-membrane and lipopolysaccharide (LPS) extracts. Histamine release from mast cells or basophils was not elicited by any of these bacterial preparations alone. However, when mixed with various secretory stimulants, the bacterial preparations caused inhibition of histamine release from rat mast cells (calcium ionophore A23187, compound 48/80, concanavalin A, anti-rat IgE) and human basophils (A23187, N-formyl Met-Leu-Phe). The degree of inhibition ranged from 48 % to 97 %. These results indicate that H. pylori exerts an inhibitory effect on cells of the immune system that contributes to its persistence within the gastric mucosa.

Treatment of intestinal Behçet's syndrome with chimeric tumour necrosis factor alpha antibody

Few patients with Behçet's syndrome have gastrointestinal ulceration. Such patients are difficult to treat and have a higher mortality. Faced with refractory symptoms in two patients with intestinal Behçet's, we used the tumour necrosis factor alpha (TNF-alpha) monoclonal antibody infliximab to induce remission. Both women (one aged 27 years, the other 30 years) presented with orogenital ulceration, pustular rash, abdominal pain, bloody diarrhoea due to colonic ulceration, weight loss, and synovitis. One had thrombophlebitis, digital vasculitis, perianal fistula, and paracolic abscess; the other had conjunctivitis and an ulcer in the natal cleft. Treatment with prednisolone, methyl prednisolone, and thalidomide in one and prednisolone, colchicine, and cyclosporin in the other was ineffective. After full discussion, infliximab (3 mg/kg, dose reduced because of recent sepsis in one, and 5 mg/kg in the other) was administered. Within 10 days the ulcers healed, with resolution of bloody diarrhoea and all extraintestinal manifestations. A second infusion of infliximab was necessary eight weeks later in one case, followed by sustained (>15 months) remission on low dose thalidomide. Remission was initially sustained for 12 months in the other but thalidomide had to be stopped due to intolerance, and a good response to retreatment lasted only 12 weeks without immunosuppression, before a third infusion. The cause of Behçet's syndrome is unknown but peripheral blood CD45 gammadelta T cells in Behçet's produce >50-fold more TNF-alpha than controls when stimulated with phorbol myristate acetate and anti-CD3. Infliximab could have a role for inducing remission in Behçet's syndrome.

Rapid serological diagnosis of Helicobacter pylori:a need for caution and re-evaluation

Screening for Helicobacter pylori in dyspeptic patients may improve selectivity for gastroscopy. Rapid serological tests based on ELISA technique are cheap, readily available and simple to use in the clinical setting. However local evaluation is essential in order to validate these techniques. Fifty-six dyspeptic patients (aged less than 45 yr) had a rapid serological test (Helisal) performed prior to gastroscopy. At gastroscopy H. pylori status was assessed using culture and histology. The Helisal sensitivity was 80 per cent, specificity 82 per cent. Screening patients with the Helisal test would have missed 6 patients with peptic ulcer disease and 2 with oesophagitis. The Helisal test did not perform satisfactorily as a screening test in selection of patients for gastroscopy.

Open access gastroscopy--3 year experience of a new service

We have audited the first 3 yr of a new open access gastroscopy service in the Royal Victoria Hospital, Belfast to assess service demands, patient demography and diagnostic trends. Over 3 yr there were 1872 referrals (800 from fundholding general practitioners), 8.8 per cent were non attenders and 5.4 per cent cancelled appointments. Endoscopic diagnostic categories showed no significant change over the 3 yr, 39 per cent non ulcer dyspepsia, 35 per cent gastro-oesophageal reflux disease (GORD), 17 per cent peptic ulcer disease (PUD), 6 per cent GORD and PUD, 1 per cent gastric erosions and 0.8 per cent carcinoma.

An audit of hospital admissions for acute upper gastrointestinal haemorrhage

A retrospective survey was made of all 189 patients admitted with acute upper gastrointestinal haemorrhage to the Belfast City Hospital in one year. The commonest single reason for admission was peptic ulcer disease, but this was lower than in other published series from the United Kingdom. Overall mortality was 4.8%. The majority of patients did not require either blood transfusion or surgery. There may be potential benefits of endoscopic haemostatic techniques to deal with this condition.

Prophylactic ranitidine treatment in critically ill children - a population pharmacokinetic study

Aims: To characterize the population pharmacokinetics of ranitidine in critically ill children and to determine the influence of various clinical and demographic factors on its disposition. Methods: Data were collected prospectively from 78 paediatric patients (n = 248 plasma samples) who received oral or intravenous ranitidine for prophylaxis against stress ulcers, gastrointestinal bleeding or the treatment of gastro-oesophageal reflux. Plasma samples were analysed using high-performance liquid chromatography, and the data were subjected to population pharmacokinetic analysis using nonlinear mixed-effects modelling. Results: A one-compartment model best described the plasma concentration profile, with an exponential structure for interindividual errors and a proportional structure for intra-individual error. After backward stepwise elimination, the final model showed a significant decrease in objective function value (-12.618; P <0.001) compared with the weight-corrected base model. Final parameter estimates for the population were 32.1lh for total clearance and 285l for volume of distribution, both allometrically modelled for a 70kg adult. Final estimates for absorption rate constant and bioavailability were 1.31h and 27.5%, respectively. No significant relationship was found between age and weight-corrected ranitidine pharmacokinetic parameters in the final model, with the covariate for cardiac failure or surgery being shown to reduce clearance significantly by a factor of 0.46. Conclusions: Currently, ranitidine dose recommendations are based on children's weights. However, our findings suggest that a dosing scheme that takes into consideration both weight and cardiac failure/surgery would be more appropriate in order to avoid administration of higher or more frequent doses than necessary.