Isolation identification and quantitative culture of Helicobacter pylori from gastric mucosa

The biopsy specimens were homogenized and diluted ten-fold serially in Hank's solution. Each dilution samples were streaked on Skirrow's selective medium. The plates were incubated at 37 degrees C 5 to 7 days under micro-aerobic condition and counting the viable cell of H. pylori. The positive rate of specimens with H. pylori in active gastric ulcer tissue (62.0%) was higher than that in scarred tissue (51.9%). Number of H. pylori viable cells in active ulcer tissue was significantly larger than in scarred tissue. The positive correlation between Raw's gastritis score of specimens from patients with erosive gastritis and number of H. pylori viable cells was clearly noticed.     

Acute effects of N-methyl-N'-nitro-N-nitrosoguanidine on canine gastric mucosa

The electrophysiological effects of the chemical gastric carcinogen N-methyl-N'-nitro-N-nitrosoquanidine (MNNG) were determined in an in vivo chambered canine stomach and in an in vitro canine gastric mucosal preparation. In the in vivo stomach, the topical application of 2.5 mg MNNG/ml decreased the transmural electrical potential difference, and the systemic blood pressure was essentially unchanged. In the in vitro preparation, exposure of the mucosal side of the isolated canine gastric mucosa to 0.25 and 2.5 mg MNNG/ml for 1 hour sequentially or exposure of the serosal side to 2.5 mg MNNG/ml for 2 hours inhibited net Na+ and Cl- fluxes. With longer duration, the undirectional fluxes of Na+ and Cl- increased, indicating an increase in permeability. These findings suggested that inhibition of active transport in the gastric mucosa may have an important function in the gastric carcinogenicity of MNNG.     

Reactions from rat gastric mucosa during one year of Helicobacter pylori infection

The aim of the present study was to investigate responses from the gastric mucosa of rats during long-term H. pylori infection. Twenty-four Sprague-Dawley rats were inoculated with a mouse-adapted strain of human H. pylori (vacA+, cagA+), 16 uninfected rats served as controls. Three to six rats from each group were killed two weeks or two, six, or 12 months later. At sacrifice, blood was sampled and the gastric mucosa was taken for bacterial culture, histology, immunocytochemistry and in situ hybridization. H. pylori colonized the antrum in 23/24 inoculated rats; with time the density of bacteria increased. The inflammation in the antral mucosa was mild to moderate and was dominated by infiltration of lymphocytes and macrophages. Serum H. pylori-specific IgG2a was significantly increased in the infected rats. The frequency of epithelial cell apoptosis was significantly increased in the early months of infection. The mucosal expression of trefoil peptide mRNA remained unchanged. We conclude that after one year of H. pylori infection in rats, the mucosal responses were rather mild, indicating that the animals may adapt to the infection by mechanisms which remain to be identified.     

Analysis of cell damage and proliferation in Helicobacter pylori-infected human gastric mucosa from patients with gastric adenocarcinoma

Helicobacter pylori (HP) infection, a cause of multifocal atrophic gastritis, is considered an important factor related to the evolution of the human gastric mucosa from normal to intestinal-type adenocarcinoma. We examined cell proliferation and both double and single strand DNA damage in situ in 35 patients undergoing gastrectomy for adenocarcinoma with HP-infected gastric mucosa by immunolocalization of Ki-67, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling, and in situ nick translation. We also studied the distribution of intraepithelial neutrophils by elastase immunolocalization. HP infection was confirmed in all cases by serum anti-HP antibodies, ureas testing, and histopathological examination. HP-infected gastric mucosa was classified according to the degree of inflammation and intestinal metaplasia. Ki-67, terminal deoxynucleotidyl transferase-mediated labeling, in situ nick translation, and intraepithelial neutrophil indices all increased with the progression of gastritis and were highest in glands with incomplete intestinal metaplasia. All indices were lowest in gastric glands with complete intestinal metaplasia. Significant positive correlations were observed among these markers. Increased proliferative activity in HP-associated chronic gastritis in response to cell damage or injury was clearly demonstrated, suggesting that both HP-associated toxins and intraepithelial neutrophils are important in HP-related gastric epithelial injury. Increased cell turnover associated with incomplete intestinal metaplasia may result in DNA instability and subsequent development of intestinal-type gastric adenocarcinoma in HP-infected mucosa.     

Polyamine profile in human gastric mucosa infected by Helicobacter pylori

Septicemia is a frequent cause of death in HIV-infected adults in developing countries. Additional prospective studies are needed to determine the etiology of bloodstream infections (BSI) in febrile HIV-infected adults and guide initial evaluation and treatment in this setting. We assessed the prevalence and etiology of community-acquired BSI among 299 consecutive febrile adult medical admissions to Mulago Hospital, Kampala, Uganda, over a 4-month period in 1997. The median age of our patients was 30 years, 159 (53%) were male, and 227 (76%) HIV-1-seropositive. Overall, prevalence of bacteremia or fungemia (1 patient) was 24%. Bacteremia was more frequent in HIV-infected than in uninfected patients (27% versus 15%, respectively; p = .04). Mycobacterium tuberculosis (n = 28), Streptococcus pneumoniae (n = 15) and Salmonella species (n = 13) were the most frequent isolates. All Salmonella and mycobacterial isolates were recovered from HIV-infected patients. Pneumococcal bacteremia was not associated with HIV seropositivity. M. avium complex and M. simiae were isolated from two HIV-infected patients. The rate of mycobacteremia among febrile HIV-infected adults presenting for hospitalization was 13%. Bacteremia and disseminated tuberculosis are frequent causes of morbidity in febrile HIV-infected Ugandan adults. Initial empiric antibiotic coverage in this setting should be targeted toward the pneumococcus and gram-negative enteric bacilli, especially nontyphi Salmonella species. All patients presenting with chronic cough should be evaluated for tuberculosis.     

Changes in gastric mucosa and Helicobacter pylori infection in young health volunteers

The effect of microinjection of a GABAB receptor agonist, baclofen, into the ventral tegmental area on the rewarding effect of morphine was investigated using the conditioned place preference paradigm in rats. Morphine (1-8 mg/kg, s.c.) caused a dose-related place preference for the drug-associated place. In contrast, microinjection of baclofen (0.1-1 nmol/side) into the ventral tegmental area did not produce a significant preference for either compartment of the test box. Pretreatment with baclofen (0.1-1 nmol/side) into the ventral tegmental area dose dependently suppressed the morphine (8 mg/kg, s.c)-induced place preference. This suppression of the morphine (8 mg/kg, s.c.)-induced place preference by baclofen (1 nmol/side), but not with the GABAA receptor antagonist bicuculline (1 nmol/side). The present results suggest that a decrease in GABAB neurotransmission in the ventral tegmental area, which may be produced via inhibition of a tonic GABAergic input by morphine, may be involved in the expression of the rewarding effect of morphine.     

Isolation rate of Enterococcus spp. from surgical infections and their susceptibilities

Enterococcus spp. isolated from surgical infections during the period from July 1982 to June 1995 were investigated in a multicenter study involving 19 hospitals in Japan, and the following results were obtained. 1. Though the isolation rate of Enterococcus faecalis and other Enterococcus spp. were not high from primary infections, and from postoperative infections the isolation rate of other Enterococcus spp. was also low, the isolation rate of E. faecalis was highest from postoperative infections after 1993. 2. Vancomycin (VCM) showed strongest activity against E. faecalis, and followed by those of ampicillin (ABPC), imipenem. levofloxacin (LVFX) and meropenem in this order. Against other Enterococcus spp., VCM showed strongest activity, and followed by those of ABPC and LVFX. There were no resistant strains against VCM.     

Hemorrhage from acute ulcers of gastric mucosa

The work analyses the results of management of 429 patients with acute erosive-ulcerative gastritis in a specialized department. The most frequent causes are duodeno- or enterogastric reflux (24.7%), abuse of alcohol (23.6%), medication with mucosa irritating drugs (22.5%). Particular attention is drawn to gastritis of a reflux character which is dangerous not only because of recurrent bleeding but also because of the possibility of structural reorganization of the gastric mucosa leading to development of carcinoma. Attention is focused on the errors in nonoperative and surgical treatment and the need for taking into consideration the etiopathogenesis of acute ulcers. In ulcerations of a reflux character it is recommended that after bleeding is arrested by conservative measures, an operation for removal of the reflux is performed in a planned order. In the other cases, in emergency situations, deep stitching of the erosions with vagotomy and pyloroplasty must be resorted to. The author considers acute ulcers of the stomach to be a precancer marker and believes it necessary to register all patients for regular medical control.     

Effect of bile on lysosomal stability in the mucosa of the canine gastric antrum

The bound and free acid hydrolases from bile and bile- then acid-treated canine antral mucosa were compared with control tissue, using explants held in a series of Lucite chambers. Normal levels of activity of bound mucosal enzymes decreased as a result of bile acid insult, but "free" activity in the cytosol increased markedly. Organelle membrane stability was "tested" by submitting isolated particulate matter containing lysosomes, to hypoosmotic shock and to mechanical stress. Release of cathepsin D, only, was observed after bile treatment in vivo. Bile, followed by acid, however, damaged organelle integrity causing release or "leakage" of seven acid hydrolases.     

The morphological diagnosis of Helicobacter pylori in the gastric mucosa in chronic gastritis]

JC virus (JCV), a human polyomavirus, is the agent of the demyelinating disease progressive multifocal leukoencephalopathy (PML). JCV exists in four main genotypes in the USA. Type 1, including subtypes Type 1A and Type 1B, makes up about 64% of strains in the USA and is thought to be of European origin. Type 2 is found in Asia, and Type 3 in Africa. A fourth type is found only in the USA. In general, these genotypes differ in 1-2.5% of their DNA sequence. Thirty MS patients and 30 paired controls from Budapest were studied. The clinical course of MS was mainly secondary progressive, and patients were stable at the time of testing. Most of the controls were relatives of the probands: a spouse, parent, or child. Overall, 25 of 60 (42%) of the urines tested positive for JCV by PCR. These included 13 of 30 MS patients, and 12 of 30 controls. Genotyping in the VPI gene showed all 25 JCV strains to be Type 1. Among the MS patients, seven were Type 1A and six were Type 1B. Among the controls, nine were Type 1A and three were Type 1B. In five pairs of MS patients and controls, both were positive for JCV by PCR. Two of these were husband/wife pairs of which one pair was matched for subtype (both Type 1A), and the other was not. Two of them were mother/daughter pairs, and both were matched for subtype (both Type 1B). These findings demonstrate that JCV Type 1 predominates among Hungarians, and suggest that parent/child pairs can be used to trace JCV transmission within the MS family.     

Effect of prolonged staining with hematoxylin on detecting Helicobacter pylori in hematoxylin-eosin-stained gastric mucosa

Rab proteins are small GTP-ases localized to distinct membrane compartments in eukaryotic cells and regulating specific steps of intracellular vesicular membrane traffic. The Rab7 protein is localized to the late endosomal compartment and controls late steps of endocytosis. We have isolated, by library screening, the 5' region, including the promoter, of the mouse Rab7 gene and a Rab7 pseudogene. We have mapped, by genetic linkage analysis, the mouse Rab7 gene on Chromosome (Chr) 6 and the Rab7-ps1 pseudogene on Chr 9, where the Rab7 gene has been previously reported to map. By radiation hybrid mapping, we have located the human RAB7 gene on Chr 3, in a region homologous to the mouse Chr 6, where the Rab7 gene maps.     

Isolation of Campylobacter spp. from slaughtered cattle and swine on blood-free selective medium

The charcoal-cefazolin-sodium deoxycholate agar (CCDA) was compared with Butzler agar for selectivity of campylobacters from fecal samples. A total of 279 samples of cecal contents from 176 cattle and 103 pigs slaughtered in Saitama were examined. Fifty-five (31.3%) cattle were found to contain campylobacters by the direct plating culture on CCDA plates, compared with 29 (16.5%) on Butzler agar plates. The positive rates of pig samples on CCDA and Butzler agar were very high, being 96 (93.2%) and 86 (83.5%), respectively. After enrichment culture of 107 cattle specimens in Preston, CEM or Bu 10 broth, the isolation rate on CCDA was also higher (p < 0.01) than that on Butzler agar after enrichment in any medium. These results indicate that CCDA medium is easy to handle without blood and, as to this study shows, is superior to Butzler agar for isolation of Campylobacter species from fecal samples.     

Isolation and characterization of anti morphine analgesic peptide from canine brain

The anti-morphine peptide from canine brain, which is homologous to alpha subunit of hemoglobin of canine, has been isolated and characterized. The canine brain was homogenized and extracted in 0.1 M acetic acid. The supernatant was collected, and purified by chromatography of Sephadex G-50 and ion-exchange and RP-HPLC. A polypeptide was then obtained, which is termed as AMP5. It shows prominent antimorphine analgesic activity. AMP5 has an apparent molecular mass of 10.23kD. The isoelectric point was estimated to be around pH 6.7 by IEF. The amino acid composition of AMP5 and sequence of the N-terminal 50 amino acid residues was determined.     

Local immune response in Helicobacter pylori-infected cats and identification of H. pylori in saliva, gastric fluid and faeces

Helicobacter pylori-infected cats were screened by culture and polymerase chain reaction (PCR) for the presence of H. pylori in salivary secretions, gastric juice, gastric tissue and faeces. H. pylori was cultured from salivary secretions in six of 12 (50%) cats and from gastric fluid samples in 11 of 12 (91%) cats. A 298 base pair polymerase chain reactions (PCR) product specific for an H. pylori 26000 MW surface protein was amplified from dental plaque samples from five of 12 (42%) cats and from the faeces of four of five (80%) cats studied. Analyses of serum and mucosal secretions by enzyme-linked immunosorbent assay (ELISA) revealed an H. pylori-specific immunoglobulin G (IgG) response, and elevated IgA anti-H. pylori antibody levels in salivary and local gastric secretions. Immunohistochemical analyses of gastric tissue revealed the presence of IgM+ B cells assembled into multiple lymphoid follicles surrounded by clusters of CD4+ and CD8+ T cells. The lamina propria also contained single cells or aggregates of IgA+ and IgM+ B cells. These observations show that H. pylori can be identified in feline mucosal secretions, and that a localized IgA immune response develops in gastric tissue of H. pylori-infected cats. The findings suggest a zoonotic risk from exposure to personnel handling H. pylori-infected cats in vivaria.     

Isolation and identification of bromfenac glucoside from rat bile

Bromfenac (Duract(R)), a drug approved for pain, was expected to be metabolized by the rat to an acyl glucuronide, a metabolite formed with most compounds of similar structure. During the investigation of metabolite profiles in rat bile following administration of 1 mg/kg iv doses of 14C-bromfenac, an acid-labile metabolite was found that degraded to form 14C-bromfenac. Isolation and characterization of this metabolite indicated that it is an unusual conjugate, bromfenac N-glucoside.     

Isolation and identification of Aeromonas species from human stools

One thousand and three diarrhoeal stool samples were processed in our laboratory during the period 1996/1997 for the presence of enteric pathogens especially Aeromonas spp., which has emerged as a new agent causing diarrhoea. Ampicillin sheep blood agar was found to be the best medium for the isolation of Aeromonas spp. from stool specimens. Enteric pathogens were found in 200 (20%) stools, of which Aeromonas spp. was the second commonest pathogen isolated amounting to 21% of isolates. This study clearly indicates that Aeromonas spp. must be looked for in every diarrhoeal stool samples, specially in children below 10 years of age. Isolation and identification is cost effective and easy, if the given protocol is observed.