Kazuyoshi Maejima

Effect of oral antibiotics and bacterial overgrowth on the translocation of the GI tract microflora in burned rats.

Infections in burned patients have generally been considered to arise from exogenous organisms. Consequently, the therapy of burned patients has emphasized the use of infection control policies and topical antimicrobial agents to reduce bacterial colonization. Even though enteric bacteria are frequently found in the burn wound little attention has been paid to the patients own GI tract microflora as a potential source of organisms colonizing the burn wound. The current experiments were carried out to determine if the bacteria present in the GI tract of healthy animals would penetrate (translocate) through the GI mucosa and spread to visceral organs after a moderate or major thermal injury. The results of these experiments indicated that bacteria can translocate across the wall of the GI tract and survive in the mesenteric lymph nodes in healthy rats. Furthermore, when the GI tract microflora is altered, either due to bacterial overgrowth or under the influence of oral antibiotic therapy, not only will bacteria translocate to the mesenteric lymph nodes but bacteria will also spread to other visceral organs. The results of these experiments support the hypothesis that the GI tract can serve as a reservoir for nosocomial infections in the burned patient, since bacteria can translocate across the mucosal barrier of the GI tract after thermal injury and survive in visceral organs before colonization of the burn wound occurs.

Expression of the rat calmodulin gene II in the central nervous system: a 294-base promoter and 68-base leader segment mediates neuron-specific gene expression in transgenic mice

Deletion analysis of the rat CaMII promoter demonstrated that the segment from -294 to +68 bases of CaMII was efficient as a promoter in NIH3T3 by transient assay. We developed transgenic mice carrying a fusion gene of this promoter segment and a beta-galactosidase reporter gene. This short CaMII promoter mediated the transgene expression in pyramidal cells of the cerebral neocortex, the pyriformcortex and the hippocampal regions CA1 to CA3, in granule cells of the dentate gyrus, in Purkinje cells of the cerebellum, and in neurons of the lateral vestibular nucleus of pons and the spinal cord of adult transgenic mice. The expression of endogenous CaMII was precisely analyzed by in situ hybridization in the nervous tissues. The localization of transgene expression was consistent with those of the endogenous CaMII in the adult transgenic mice. In the embryos at 13.5-15.5 days of gestation, the transgene was expressed in various neurons similarly to the endogenous CaMII but certain subtle differences were observed in the localization of expression. This short promoter of rat CaMII carried two sequence stretches highly conserved in the mouse, dog, chicken and Xenopus CaMII promoters. These conserved stretches may be involved in the observed neuron-specific expression of rat CaMII gene.

Translocation of bacteria from the gastrointestinal tract in immunodeficient mice

Host defence mechanisms associated with the inhibition of translocation of bacteria from the gastrointestinal (GI) tract were investigated in SCID and beige mice after decontamination with oral antibiotics and colonization with Escherichia coli C25. SCID mice, which have impaired T and B cell function, tended to have a greater incidence of bacterial translocation from the GI tract up to 7 days after inoculation compared with controls. However, after 7 days both SCID and controls cleared the E. coli C25 from the liver, spleen, blood and peritoneal cavity. Beige mice, with impaired NK cell and polymorphonuclear leukocyte function, were not able to clear the inoculated bacteria from their liver by 14 days after inoculation although the controls were cleared by 7 days. Numbers of bacteria in the mesenteric lymph nodes (MLN) of beige mice did not decrease significantly by 14 days after inoculation, whereas numbers in SCID mice decreased markedly within 7 days. These results suggest that defence mechanisms other than T and B cell function are important in the inhibition of systemic infection from the GI tract.

Stability of pathogenic bacteria from laboratory animals in various transport media

The stability of pathogenic bacteria from laboratory animals was investigated in various transport media at different temperatures. Bordetella bronchiseptica and Salmonella typhimurium survived for 8 days in phosphate-buffered saline (PBS, pH 7·0) at 37, 24, 4 and -20°C; Brucella canis at 24, 4 and -20°C; Corynebacterium kutscheri at 4 and -20°C; and Pseudomonas aeruginosa at all but -20°C. A marked decrease in numbers of Pasteurella multocida and Past. pneumotropica was observed in PBS at all temperatures. Skimmed milk in PBS improved the survival of Pasteurella spp. and Ps. aeruginosa at -20°C. Neither glycerin, ascorbic acid nor sodium thioglycollate improved survival. The numbers of viable B. canis, Ps. aeruginosa and S. typhimurium were maintained in blood or faecal specimens held for 8 days at 4°C. These results indicated that transport in PBS at 4°C was the only method satisfactory for all species of pathogenic organisms tested, but Pasteurella spp. were the most difficult to maintain.

Bacterial Translocation from the Gastrointestinal Tract in Various Mouse Models for Human Diabetes

Bacterial translocation from the gastrointestinal (GI) tract to other internal organs was examined in multiple low‐dose streptozotocin‐injected (M‐STZ), single large‐dose streptozotocin‐injected (S‐STZ), alloxan‐injected (Alloxan), and non‐obese diabetic (NOD) mice. The incidence of bacterial translocation from the GI tract to the tested organs among diabetic mice was in the order of M‐STZ mice>S‐STZ mice>NOD, Alloxan, and control mice. The injections of insulin to M‐STZ mice did not decrease the incidence of translocation. These results suggest that bacterial translocation from the GI tract in diabetic mice is not induced by diabetes.