HeLa cell invasiveness and O antigen of Shigella flexneri as separate and prerequisite attributes of virulence to evoke keratoconjunctivitis in guinea pigs.

Many rough mutants selected from isogenic smooth virulent and avirulent strains of Shigella flexneri were examined for virulence, using tissue culture infection and Sereny tests. Many of the rough mutants isolated from a virulent smooth strain were capable of penetrating tissue culture cells but incapable of producing a positive Sereny test. In contrast, we could not obtain from smooth avirulent strains any rough mutants capable of penetrating HeLa cells. Chemical analysis of lipopolysaccharide of some representative rough strains showed several patterns of sugar composition with a range of from Ra to Re chemotypes. There was no correlation between HeLa cell invasiveness and chemotypes of lipopolysaccharides, thus indicating little significance of oligosaccharides of the rough core as well as O antigens in the ability of S. flexneri to penetrate HeLa cells. When these invasive rough strains were given O antigen genes from a smooth avirulent Shigella Hfr strain, most of the transconjugants that expressed O antigens regained the ability to evoke keratoconjunctivitis in guinea pigs. We also examined the chromosomal loci of HeLa cell invasion by transferring carbohydrate fermentation genes of Escherichia coli K-12 Hfr and found two chromosomal loci, the rha and lac-gal regions, which control the ability to penetrate HeLa cells. These results suggested that O antigens and ability to penetrate tissue culture cells are independent and prerequisite attributes of virulence in Shigella flexneri to evoke keratoconjunctivitis in guinea pigs.     

IL-12 p40 prevents the development of chronic enterocolitis in IL-10-deficient mice

T-helper-1 (Th1) cytokines play an important role in Crohn's disease, and interleukin-12 (IL-12), which is composed of two subunits, p40 and p35, drives Th1 differentiation. In previous reports, IL-12 p40 was shown to prevent IL-12 from binding to the receptor. We demonstrate here the effect of IL-12 p40 overexpression in intestinal epithelia on enterocolitis mediated by Th1 cells in IL-10-deficient (IL-10(-/-)) mice on a C57BL/6J background. IL-10 deficient (IL-10(-/-))/T3b-IL-12 p40+ (IL-12 p40+) mice and IL-10(-/-)/T3b-IL-12 p40- (IL-12 p40-) mice were generated by crossing T3b-IL-12 p40 transgenic mice and IL-10(-/-) mice. At 8 weeks of age, IL-12 p40+ mice did not show any clinical manifestations of colitis. The colon length of IL-12 p40- mice became shorter than that of IL-12 p40+ mice. The histological score of IL-12 p40+ mice was lower. Interferon-gamma (IFN-gamma) production was suppressed in both the mesenteric lymph node cell culture and colon tissue culture of IL-12 p40+ mice. There was no significant difference in IL-4 production and tumor necrosis factor-alpha (TNF-alpha) production between the two groups. These results show that overexpression of IL-12 p40 in intestinal epithelia prevents enterocolitis in IL-10(-/-) mice by suppressing IFN-gamma production, and suggest a potential clinical application of IL-12 p40 for Crohn's disease. Furthermore, these results also suggest that local gene transduction in the intestinal epithelium may be a potent therapeutic approach for Crohn's disease.     

Ras homolog gene family H (RhoH) deficiency induces psoriasis-like chronic dermatitis by promoting TH17 cell polarization

1. Download : Download high-res image (104KB)
2. Download : Download full-size image

     

Primary structure,functional characterization and developmental expression of the ascidian Kv4-class potassium channel

Ascidians belong to the primitive chordates and their larvae show symmetrical beating of the tail, which is reminiscent of the swimming pattern in primitive vertebrates. Since ascidian larva contains only a small number of neurons in their entire larval nervous system, they will potentially provide a simple model for the study of animal locomotion. In a step towards the goal of establishing the molecular basis underlying ascidian larval neurophysiology, we describe here a Kv4 class of voltage-gated potassium channel, TuKv4, from Halocynthia roretzi. Whole mount in situ hybridization indicates that TuKv4 is expressed in most of larval neurons including motor neurons. TuKv4-currents reconstituted in Xenopus oocytes show currents with similar properties to the lower-threshold A-type currents from cleavage-arrested ascidian blastomeres of neural lineage. However, the voltage-dependency of the steady-state inactivation and activation was shifted leftward by 20 mV, as compared with native A-type currents, suggesting that other components may be required to restore full function of the Kv4 channel. Unexpectedly, another isoform lacking C-terminal cytoplasmic region was also isolated. This truncated isoform did not lead to a functional current in Xenopus oocytes. RT-PCR analysis showed that the truncated form is transiently expressed during larval development, suggesting some developmental role for potassium channel expression.     

Reproduction of menstrual changes in transplanted human endometrial tissue in immunodeficient mice

BACKGROUND: Cultures of human endometrial tissue are useful for analysing the mechanisms underlying the menstrual cycle. However, long-term culture of endometrial tissue is difficult in vitro. Xenotransplantation of normal human endometrial tissue into immunodeficient mice could allow prolonged survival of the transplanted tissues. METHODS: Proliferative-phase endometrial tissue samples from three women were transplanted into the subcutaneous space of ovariectomized, immunodeficient, non-obese diabetic (NOD)/severe combined immunodeficiency (SCID)/gammaC(null) (NOG) mice. The mice were treated with 17beta-estradiol (E2) for the first 14 days after transplantation, followed by E2 plus progesterone for the next 14 days. The transplants were investigated morphologically and immunohistochemically at various times after implantation. RESULTS: The transplanted tissues contained large numbers of small glands, pseudostratification of the nuclei and dense stroma after treatment with E2 alone. After treatment with E2 plus progesterone, subnuclear vacuolation, luminal secretion and decidualization of the stroma were observed. When the hormone treatment ceased, tissue destruction occurred and the transplants returned to the proliferative phase. Lymphocytes were identified immunohistochemically: the numbers of CD56-positive and CD16-negative cells increased significantly in the stroma during the late secretory phase (day 28). CONCLUSIONS: Human endometrial tissue transplanted into NOG mice showed similar histological changes to eutopic endometrial tissue during treatment with sex steroid hormones for 1 month. Moreover, lymphocytes were produced in the transplanted human endometrial tissue. This system represents a new experimental model of the human endometrium in vivo.     

Mapping nondominant voices into understanding stress-coping mechanisms

This study reports key findings from a research project, which examined the stress and coping mechanisms of several nondominant groups of individuals. The groups were based in Winnipeg, Manitoba, Canada and included (a) Aboriginal individuals with diabetes, (b) individuals with disabilities, and (c) gays and lesbians. Our analyses of personal narratives and life stories have led to develop an interpretive map of findings that depicts mechanisms of how stress and coping operate. Specifically, the interpretive map consists of personal and structural stressors, meanings of stress, and personal and structural resources, as well as of two constructs termed intersectionality and social exclusion. Not only are nondominant voices and lived experiences recognized and incorporated into an emergent interpretive map, but this map also articulates the complex ways in which multiple identities intersect (i.e., intersectionality) and the realities of being excluded socioeconomically, culturally, and politically among nondominant groups (i.e., social exclusion). © 2008 Wiley Periodicals, Inc.     

Avoidance learning under delayed shock termination in prenatally x-irradiated rats

Groups of MP1 male albino rats were exposed to x-irradiation of 200 rad on Day 17 of gestation. Irradiated and control rats were trained in a shuttlebox with either immediate or .5-sec delayed termination of both the electric shock (US) and the conditioned stimulus (CS) on escape trials. An avoidance response always produced prompt CS termination. Irradiated rats showed a higher degree of avoidance behavior than controls and, under the delay condition, tended to run rapidly in the CS-US interval.