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[Pathogenesis of Helicobacter pylori infection--bacterium and host relationship].

Helicobacter pylori (H. pylori) colonizes the gastric mucosa of a half of the mankind. Duodenal ulcer is found in 15-25%, gastric ulcer in 13%, while gastric adenocarcinoma develops in 1% of all infected individuals. Pathogenesis of H. pylori infection is related to the virulence factors of the bacterium, environmental (dietary habits, hygiene, stress) and host factors (age, sex, blood type). Colonization of the gastric mucosa is related to the motility of the bacterium, presence of lipopolysaccharide (LPS) and various bacterial enzymes. Gastric mucosal injury is the result of H. pylori LPS, vacuolization cytotoxin (vacA), cytotoxin associated protein (cagA), heat shock proteins and factors responsible for neutrophil chemotaxis and activity. H. pylori colonizes the gastric mucosa and zones of ectopic gastric epithelium. H. pylori infection is transmitted via oral-oral, fecal-oral and iatrogenic way (during endoscopy). Higher prevalence of the infection is associated with lower socioeconomic level, lack of drinking water, and living in a community. Acute H. pylori gastritis is superficial pangastritis progressing into the chronic phase after 7-10 days. Gastric mucosal atrophy and intestinal metaplasia can develop during the course of H. pylori infection. Clearly defined factors that influence the outcome of H. pylori infection include bacterial strain, distribution of gastritis, acid secretion and gastric mucosal atrophy.

Duodenum↗

Diaphorobacter nitroreducens gen nov, sp nov, a poly(3-hydroxybutyrate)-degrading denitrifying bacterium isolated from activated sludge.

Three denitrifying strains of bacteria capable of degrading poly(3-hydroxybutyrate) (PHB) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) were isolated from activated sludge and characterized. All of the isolates had almost identical phenotypic characteristics. They were motile gram-negative rods with single polar flagella and grew well with simple organic compounds, as well as with PHB and PHBV, as carbon and energy sources under both aerobic and anaerobic denitrifying conditions. However, none of the sugars tested supported their growth. The cellular fatty acid profiles showed the presence of C16:1omega7cis and C16:0 as the major components and of 3-OH-C10:0 as the sole component of hydroxy fatty acids. Ubiquinone-8 was detected as the major respiratory quinone. A 16S rDNA sequence-based phylogenetic analysis showed that all the isolates belonged to the family Comamonadaceae, a major group of beta-Proteobacteria, but formed no monophyletic cluster with any previously known species of this family. The closest relative to our strains was an unidentified bacterium strain LW1 (=DSM 13225) (99.9% similarity), reported previously as a 1-chloro-4-nitrobenzene degrading bacterium. DNA-DNA hybridization levels among the new isolates were more than 60%, whereas those between our isolates and strain DSM 13225 were less than 50%. The G+C content of genomic DNA of the new strains was 64 to 65 mol%. Based on these results, we concluded that the PHBV-degrading denitrifying isolates should be classified as a new genus and a new species, for which we propose the name Diaphorobacter nitroreducens. The type strain is strain NA10B (=JCM 11421=CIP 107294). We also propose to classify strain DSM 13225 as a genospecies of Diaphorobacter.

Bacterial Typing Techniques↗

Comamonas badia sp. nov., a floc-forming bacterium isolated from activated sludge.

Strain IAM 14839, isolated from activated sludge in Japan, forms a visible floc and grows in the flocculated state. This bacterium is Gram-negative, rod-shaped, strictly aerobic and highly motile with a single polar flagellum. Both oxidase and catalase activities are positive. No growth was observed on sugars. The strain can grow at 20 degrees C, but does not grow at 37 degrees C. The G+C content of DNA is 66.3 mol% and Q-8 is the major quinone. The major cellular fatty acids are 16:1omega7c, 16:0, 18:1omega7c, 2OH 16:0, 3OH 10:0. The 16S rDNA sequence analysis indicated that the bacterium clustered within the genus Comamonas. On the basis of the phylogenetic analysis and phenotypic properties, it is proposed that the strain IAM 14839T be classified in a novel species of the genus Comamonas, Comamonas badia sp. nov. The type strain is IAM 14839T (=KCTC 12244T ).

Base Composition↗

Growth, survival, and resistance of the Legionnaires' disease bacterium.

The Legionnaires' disease (LD) bacterium can now be readily cultured on artificial media. Studies were done to define the growth and survival of the LD bacterium in these media and ascertain its susceptibility to disinfecting agents. Growth-curve studies of the Philadelphia 1 strain using Mueller-Hinton broth with ferric pyrophosphate and L-cysteine (Feeley-Gorman broth) showed a lag phase of less than 24 h, a generation time of 3.8 h during the logarithmic phase, a plateau of 2 x 10(7) organisms per millilitre, and continued viability for as long as 110 d. Viability on chocolate agar with 1% hemoglobin and 2% IsoVitaleX added reached 150 d. This strain was susceptible to a variety of commonly recommended hospital and laboratory disinfectants, often in low concentrations. These investigations suggest that prolonged survival may occur in natural as well as artificial milieus and that low concentrations of phenolics, quaternary ammonium compounds, glutaraldehyde, formaldehyde, and hypochlorite could eradicate potential reservoirs for human infection.

Bacteria↗

Fine structure of the Legionnaires' disease bacterium. In-vitro and in-vivo studies of four isolates.

We obtained four bacterial isolates from patients with Legionnaires' disease and examined them for in-vitro and in-vivo fine-structure characteristics. All isolates had an outer membrane, cytoplasmic membrane, and intracellular membrane structure. Numerous intracellular inclusions were seen, particularly from in-vivo specimens, and appeared membrane-limited. Fine-structure analysis did not reveal the presence of a definitive peptidoglycan structure. Isolation, purification, and chemical analysis of Legionnaires' disease bacterium pepdoglycan established molar ratios of alanine-glutamic acid and muramic acid-glucosamine. Diaminopimelic acid was absent in the Legionnaires' disease bacterium peptidoglycan. The Kellenberger procedure for fixation appears to be the best method for the fine-structure determination of Legionnaires' disease bacteria.

Alanine↗

Isolation of the Legionnaires' disease bacterium from environmental samples.

We analyzed 24 environmental samples collected in or near the Indiana Memorial Union, where an epidemic of Legionnaires' disease occurred in early 1978. We conducted fluorescent antibody analyses and culture on F-G and charcoal yeast extract agars of each sample directly; splenic tissue of guinea pigs inoculated with the sample; and yolk sacs from embryonated eggs inoculated with splenic tissue of guinea pigs injected with the sample. Legionnaires' disease (LD) bacterium was isolated from seven of the 24 samples: one water sample from the air-conditioner cooling tower of the Union; three water samples from a stream near the Union; and three mud samples from the same stream. The LD bacterium strains were of three different serotypes. These findings indicate that LD bacteria may be widespread in nature.

Air Conditioning↗

Pathologic findings in guinea pigs inoculated intraperitoneally with the Legionnaires' disease bacterium.

Tissues from guinea pigs inoculated intraperitoneally with the Legionnaires' disease (LD) bacterium were studied with light, immunofluorescent, and electron microscopy. The principal gross lesion was diffuse peritonitis of varying severity. Microscopically, the peritonitis of covered by a mixed inflammatory infiltrate of macrophages, neutrophils, fibrin, and cellular debris. Foci of inflammation and necrosis were consistently observed in the splenic parenchyma, and similar lesions were often found in the lungs, liver, lymph nodes, pancreas, heart, and other organs. Numerous LD bacteria were seen in the peritoneal exudate; fewer were found in disseminated lesions. In electron micrographs, the highest concentrations were seen in macrophages, with fewer organisms present in neutrophils or extracellular spaces. Although the lung is the primary organ konwn to be affected by Legionnaires' disease in humans, our findings indicate that the LD bacterium is capable of dissemination.

Animals↗

A Legionella-like bacterium related to WIGA in a fatal case of pneumonia.

An unusual bacterium serologically related to a "rickettsia-like agent," designated previously as WIGA, was seen in lung tissue from a patient who died of pneumonia of unknown cause. A fluorescent antibody conjugate prepared with the WIGA organism, isolated in 1959, was used to stain the lung tissue. Enormous numbers of fluorescent bacteria in the lungs of this patient confirm the pathogenicity of this unusual bacterium.

Bacteria↗

The rickettsia-like organisms TATLOCK (1943) and HEBA (1959): bacteria phenotypically similar to but genetically distinct from Legionella pneumophila and the WIGA bacterium.

Two "rickettsia-like organisms," TATLOCK and HEBA, isolated from human blood via guinea pigs and embryonated eggs in 1943 and 1959, respectively, have been cultured on artificial media (charcoal yeast extract agar) for the first time and characterized. TATLOCK and HEBA have identical cultural, biochemical, and antigenic characteristics, as well as identical cellular fatty-acid composition and antimicrobial susceptibilities. These two bacteria have most of the cultural and biochemical characteristics of Legionella pneumophilia, and their gas-liquid chromatography cellular fatty-acid profile is similar to that of WIGA, another bacterium similar to L. pneumophila. Direct fluorescent-antibody reagents prepared for HEBA and TATLOCK gave equal high-titered reciprocal staining and were negative on 220 other bacteria, including L. pneumophila. Deoxyribonucleic acid relatedness studies, however, showed that these bacteria are not genetically related to either L. pneumophila or the WIGA bacterium.

Anti-Bacterial Agents↗

[A new, sporulating, denitrifying, mesophilic bacterium: Bacillus azotoformans N. SP. (author's transl)].

The described bacterium was isolated by enrichment culture in peptone broth inoculated with garden soil, pasteurized and then put to incubate under N2O at 32 degrees. It is a Gram-negative rod, motile with peritrichous flagella, and producing oval spores without exosporium in swollen sporangia. However, cells have the thick walls, mesosomes and persistant septa characteristic of Gram-positive bacteria. It lacks fermentative activity, does not attack carbohydrates, has complex growth requirements, and will grow anaerobically only if one of the following electron acceptors is present: NO3, NO2, N2O, S4O6, and fumarate. Nitrate, nitrite, and nitrous oxide are denitrified with production of N2. The microorganism is mesophilic, gives a positive oxidase reaction, synthesizes a type of c cytochrome, and does not hydrolyse gelatin, starch nor "Tween 80". The following enzymes are present: nitrate reductase A, respiratory nitrite reductase, tetrathionate and fumarate reductases, L-glutamate dehydrogenase, and superoxide dismutase. The following enzymes are absent: thiosulfate reductase, urease, lecithinase, arginine dihydrolase, L-alanine dehydrogenase, phenylalanine desaminase, and catalase. The GC% of its DNA is 39. The bacterium described can be considered to be a new species. We propose the name Bacillus azotoformans n. sp.

Bacillus↗

[Clinical evaluation of metallo-beta-lactamase producing bacterium].

The metallo-beta-lactamase producing bacterium are increasing in recent years. We characterized cases infected or colonized with blaIMP, which is metallo-beta-lactamase IMP-1 gene, positive gram-negative rods(GNRs). About half of the cases had malignant diseases as underliningg disease. When blaIMP positive GNRs were isolated, most of cases shows clinical signs of infection such as fever and elevation of C-reactive protein in serum. Most of strains were isolated from urinary samples and all patients from whom blaIMP positive bacterium were isolated from urinary samples were inserted urinary tract catheter. It was suggested that the insertion of catheters related to infection of blaIMP positive GNRs. No antibiotics were administered to several patients before the isolation of blaIMP positive GNRs although broad spectrum antibiotics such as carbapenems were administered to more than half of the patients. It is very important to prevent their dissemination in hospitals because most of blaIMP positive GNRs were isolated in same departments and showed identical pattern by pulsed field gel electrophoresis. And it is also important for treatment of the infectious disease due to blaIMP positive GNR to remove the catheter and improve general conditions of patients.

Bacterial Proteins↗

Regulation of nitrogenase in the photosynthetic bacterium Rhodobacter sphaeroides containing draTG and nifHDK genes from Rhodobacter capsulatus.

The photosynthetic bacteria Rhodobacter capsulatus and Rhodospirillum rubrum regulate their nitrogenase activity by the reversible ADP-ribosylation of nitrogenase Fe-protein in response to ammonium addition or darkness. This regulation is mediated by two enzymes, dinitrogenase reductase ADP-ribosyl transferase (DRAT) and dinitrogenase reductase activating glycohydrolase (DRAG). Recently, we demonstrated that another photosynthetic bacterium, Rhodobacter sphaeroides, appears to have no draTG genes, and no evidence of Fe-protein ADP-ribosylation was found in this bacterium under a variety of growth and incubation conditions. Here we show that four different strains of Rba. sphaeroides are incapable of modifying Fe-protein, whereas four out of five Rba. capsulatus strains possess this ability. Introduction of Rba. capsulatus draTG and nifHDK (structural genes for nitrogenase proteins) into Rba. sphaeroides had no effect on in vivo nitrogenase activity and on nitrogenase switch-off by ammonium. However, transfer of draTG from Rba. capsulatus was sufficient to confer on Rba. sphaeroides the ability to reversibly modify the nitrogenase Fe-protein in response to either ammonium addition or darkness. These data suggest that Rba. sphaeroides, which lacks DRAT and DRAG, possesses all the elements necessary for the transduction of signals generated by ammonium or darkness to these proteins.

ADP Ribose Transferases↗

Mechanism of removal of aflatoxin B1 by a resistant bacterium.

A bacterium resistant to aflatoxin B1 isolated from the soil was shown to remove a maximum 26% of toxin at 3 microg ml(-1). A comparison of spectrophotometric and radioisotope methods showed that a maximum of 48 hr was sufficient to remove the toxin. Radioisotope analysis showed that the radioactivity decreased in the chloroform phase while it increased in the aqueous phase during the time course of the experiment. An analysis of the supernatant of culture medium showed that the bacterium had converted aflatoxin B1 to water soluble compounds with lambda(max) of 338 and 374.

Aflatoxin B1↗

[Isolation and identification of a killing maggots bacterium].

A notably killing maggots bacterium was isolated from natural dead maggots in the manure pits in the countryside of Yancheng. Its pathogenicity was confirmed by the law of KOCK. The results of preliminary bioassay show that the pathogen can infect the larvas of greenbottle flies and other larvas of flies in a certain extent, but can't infect animals and fowls. The G + C content of its DNA is 62.46%. The hybridization ratio of its DNA and the Pseudomonas pseudoalcaligenes' (AS1.1806) is 81.2%. According to Bergey's Manual of Systematic Bacteriology Ninth edition, the strain of the bacterium was primarily identified as Pseudomonas pseudoalcaligenes.

Animals↗

[Detection of Oxalobacter formigenes in human feces and study of related genes in a new oxalate-degrading bacterium].

The first objective of the present study was to examine the presence of Oxalobacter formigenes (an oxalate-degrading bacterium in the human intestine) according to sex in a large number of Japanese. The second objective was to study the presence of three related genes in Bifidobacterium breve, which is considered to be a new oxalate-degrading bacterim. Fecal samples were collected from 55 male and 37 female healthy volunteers. O. formigenes was detected by a polymerase chain reaction (PCR) and a culture-based method. DNA was amplified by the PCR method including the site of important base sequences of each gene in order to detect oxc, frc, and OxlT. O. formigenes was present in 80% of 54 male and 62% of 34 female subjects in the PCR-based assay, while it was present in 62% of 40 male and 50% of 24 female subjects in the culture-based assay. The partial base sequences of the three related genes in B. breve were determined. The RNA polymerase-binding site in promoters and the rho-independent termination sequence were preserved in oxc and frc. In conclusion, the rate of the presence of O. formigenes was the same as in previous reports. Female subjects showed a 15% lower rate than males. B. breve is considered to be an oxalate-degrading bacterium since it was found to have oxalic acid-degrading ability and three genes involved in oxalate degradation.

Adult↗

High pressure nmr study of dihydrofolate reductase from a deep-sea bacterium Moritella profunda.

We have investigated the effect of pressure and temperature on the structural and thermodynamic stability of a protein dihydrofolate reductase from a deep-sea bacterium Moritella profunda in its folate-bound form in the pressure range between 3 and 375 MPa and the temperature range between -5 and 30 degrees C. The on-line cell variable pressure 1H NMR spectroscopy has been used to analyze the chemical shift and signal intensity in one-dimensional 1H NMR spectra. Thermodynamic analysis based on signal intensities from protons in the core part indicates that the thermodynamic stability of Moritella profunda DHFR is relatively low over the temperature range between -5 and 30 degrees C (deltaG0=15.8 +/- 4.1 kJ/mol at 15 degrees C), but is well adapted to the living environment of the bacterium (2 degrees C and 28 MPa), with the maximum stability around 5 degrees C (at 0.1 MPa) and a relatively small volume change upon unfolding (deltaV= 66 +/- 19 ml/mol). Despite the relatively low overall stability, the conformation in the core part of the folded protein remains intact up to approximately 200 MPa, showing marked stability of the core of this protein.

Adaptation, Physiological↗

[The functional role of reduced inorganic sulfur compounds in the metabolism of the microaerophilic bacterium Spirillum winogradskii].

Oxidation of reduced sulfur compounds by microaerophilic sulfur bacterium Spirillum winogradskii was found to occur only concomitantly with consumption of an organic substrate and was not linked to their utilization as electron donors in energy metabolism. No enzymes of dissimilatory sulfur metabolism were found in the cells of the sulfur bacterium oxidizing thiosulfate to tetrathionate; oxidation of thiosulfate and sulfide was caused by their reaction with reactive oxygen species (ROS), mostly H2O2 produced in the course of aerobic growth. Decreased lytic effect of ROS in the presence of thiosulfate resulted in a twofold increase in the cell yield under aerobic conditions and more efficient substrate utilization. The latter effect was caused by decreased expense of energy for the biosynthesis of oxygen-protecting polysaccharides. The stimulatory effect of thiosulfate on the growth processes was due to the activation of a number of TCA cycle enzymes producing the intermediates for constructive metabolism, especially of the NADP-dependent malic enzyme. As a result of thiosulfate-induced synthesis of SH-containing cell components, the integral antioxidative activity increased 1.5-fold.

Adenosine Triphosphate↗

[The synthesis of hydroxybutyrate and hydroxyvalerate copolymers by the bacterium Ralstonia eutropha].

The paper deals with the study of the synthesis of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) copolymers by the bacterium Ralstonia eutropha B-5786 grown under different carbon nutrition conditions (growth on carbon dioxide, fructose, and CO2-valerate and fructose-valerate mixtures). The parameters to be analyzed included the yield of biomass, the yield, synthesis rate, and composition of copolymers, the activity of the key enzymes of polyhydroxyalkanoate (PHA) synthesis (beta-ketothiolase, acetoacetyl-CoA reductase, and PHA synthase), the maximum tolerable concentration of valerate to the bacterium, and the conditions that govern the incorporation of hydroxyvalerate to copolymers. This allowed the relationship between cultivation conditions and the proportion of monomers in the copolymers to be deduced. We were able to synthesize a range of 3HB/3HV copolymers and found that the thermal characteristics and the degree of crystallinity of these copolymers depend on the molar fraction of 3HV.

Acetyl-CoA C-Acyltransferase↗