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121.
Detergent-resistant membrane microdomains facilitate Ib oligomer formation and biological activity of Clostridium perfringens iota-toxin 总被引:1,自引:0,他引:1 下载免费PDF全文
Clostridium perfringens iota-toxin consists of two separate proteins identified as a cell binding protein, iota b (Ib), which forms high-molecular-weight complexes on cells generating Na(+)/K(+)-permeable pores through which iota a (Ia), an ADP-ribosyltransferase, presumably enters the cytosol. Identity of the cell receptor and membrane domains involved in Ib binding, oligomer formation, and internalization is currently unknown. In this study, Vero (toxin-sensitive) and MRC-5 (toxin-resistant) cells were incubated with Ib, after which detergent-resistant membrane microdomains (DRMs) were extracted with cold Triton X-100. Western blotting revealed that Ib oligomers localized in DRMs extracted from Vero, but not MRC-5, cells while monomeric Ib was detected in the detergent-soluble fractions of both cell types. The Ib protoxin, previously shown to bind Vero cells but not form oligomers or induce cytotoxicity, was detected only in the soluble fractions. Vero cells pretreated with phosphatidylinositol-specific phospholipase C before addition of Ib indicated that glycosylphosphatidyl inositol-anchored proteins were minimally involved in Ib binding or oligomer formation. While pretreatment of Vero cells with filipin (which sequesters cholesterol) had no effect, methyl-beta-cyclodextrin (which extracts cholesterol) reduced Ib binding and oligomer formation and delayed iota-toxin cytotoxicity. These studies showed that iota-toxin exploits DRMs for oligomer formation to intoxicate cells. 相似文献
122.
McCaustland KA Krawczynski K Ebert JW Balayan MS Andjaparidze AG Spelbring JE Cook EH Humphrey C Yarbough PO Favorov MO Carson D Bradley DW Robertson BH 《Archives of virology》2000,145(9):1909-1918
Summary. Different patterns of disease were observed among 11 chimpanzees who were inoculated intravenously with hepatitis E virus
(HEV) positive fecal specimens from four different outbreaks (Nepal 1981, Uzbekistan 1981, Pakistan 1985, and Mexico 1986).
Five chimpanzees had marginal or no liver enzyme elevations within 70 days of inoculation. Two of the chimpanzees had limited
viremia, but did not produce detectable antibody. The four remaining chimpanzees had liver enzyme elevations, viral shedding,
viremia, seroconversion to anti-HEV, and detectable HEV antigen in liver biopsy specimens. These results may reflect the range
of infection patterns that develop in humans after natural exposure to the HEV.
Received December 29, 1999/Accepted February 21, 2000 相似文献
123.
We discuss the advantages and problems associated with fitting geometric data of the human torso obtained from magnetic resonance
imaging, with high-order (bicubic Hermite) surface elements. These elements preserve derivative (C
1) continuity across element boundaries and permit smooth anatomically accurate surfaces to be obtained with relatively few
elements. These elements are fitted to the data with a new nonlinear fitting procedure that minimizes the error in the fit
while maintainingC
1 continuity with nonlinear constraints. Nonlinear Sobelov smoothing is also incorporated into this fitting scheme. The structures
fitted along with their corresponding root meansquared error, number of elements used, and number of degrees of freedom (df)
per variable are: epicardium (0.91 mm, 40 elements, 142 df), left lung (1.66 mm, 80 elements, 309 df), right lung (1.69 mm,
80 elements, 309 df), skeletal muscle surface (1.67 mm, 264 elements, 1,010 df), fat layer (1.79 mm, 264 elements, 1,010 df),
and the skin layer (1.43 mm, 264 elements, 1,010 df). The fitted surfaces are assembled into a combined finite element/boundary
element model of the torso in which the exterior surfaces of the heart and lungs are modeled with two-dimensional boundary
elements and the layers of the skeletal muscle, fat, and skin are modeled with finite elements. The skeletal muscle and fat
layers are modeled with bicubic Hermite linear elements and are obtained by joining the adjacent surface elements for each
layer. Applications for the torso model include the forward and inverse problems of electrocardiography, defibrillation studies,
radiation dosage studies, and heat transfer studies. 相似文献
124.
Joachim Saloga Gideon Lack Kathrine Bradley Harald Renz Gary Larsen Donald Y. M. Leung Erwin W. Gelfand 《European journal of immunology》1994,24(12):3140-3147
We investigated the ability of staphylococcal enterotoxin B (SEB) to modify the immediate hypersensitivity response induced in BALB/c mice following sensitization to ovalbumin (OVA), a response mediated by OVA-reactive Vβ8 T cells. Mice were sensitized by skin painting with OVA every second day over a period of 2 weeks. SEB, a potent activator of Vβ8+ T cells, was administered at the same site where OVA was applied (skin of the lower abdomen) following two different protocols. In protocol (A) SEB was injected intradermally 1 day before painting with OVA and on day 7; in protocol B, SEB was injected each time OVA was applied to the skin (eight times). SEB (but not SEA) altered the development of immediate hypersensitivity to OVA, as demonstrated by the reduction in allergen-specific IgE, decreased OVA-specific immediate skin test responsiveness, and prevented the development of increased airways responsiveness after bronchial challenge with OVA. Injections of SEB did not alter the proliferative responses of local draining lymph node cells or spleen mononuclear cells to OVA, indicating that administration of SEB did not inhibit the sensitization to OVA, but shifted the immune response away from an immediate type response (IgE/IgG1) to IgG2a, IgG2b and IgG3. Although both protocols of SEB treatment did not lead to a major deletion of the Vβ8 T cell population, they did reduce the proliferative response of Vβ8+ T cells to OVA. These data indicate that the bacterial toxin SEB is capable of modifying the immediate hypersensitivity response induced by OVA by altering the functional capacity of antigen-reactive Vβ8 T cells. 相似文献
125.
Sinusitis is one of the most common reasons patients visit their primary care physician. The etiology of sinusitis is multifactorial
in most cases. However, the final common pathway of disruption is usually made with a thorough history. The physical examination
is often unremarkable. Complaints of nasal obstruction, mucopuluent nasal drainage, and facial pain/pressure are most diagnostic
chronic sinusitis. Isolated headache is an uncommon presenting symptom. Computed tomography scans are the gold standard for
diagnostic imaging. They can be used both for diagnosis and surgical treatment. All chronic sinusitis patients, being considered
for endoscopic sinus surgery, should have failed a trial of maximal medical therapy. This includes a 4–6 wk course of oral
antibiotics, nasal steroids, topical nasal decongestants, and oral prednisone if possible. Patients who fail maximal medical
therapy have persistent symptoms that significantly effect their daily activities, have chronic abnormalities on computet
tomography scan, and are candidates for endoscopic sinus surgery. Appropriate patient selection and preoperative counseling
are key factors in patient satisfaction. Most patients with symptoms that significantly impact their daily activities will
receive marked improvement in symptoms after sinus surgery. Endoscopic sinus surgery has undergone radical changes in the
last 15 yr. Minimally invasive techniques, combined with advances in instrumentation and computers have reduced postoperative
discomfort and improved patient satisfaction. 相似文献
126.
The nucleotide sequence of the VP1 region of marmoset-attenuated hepatitis A virus (HAV), MS-1, was determined by incorporative dideoxynucleotide sequencing of the RNA obtained from purified, liver-derived virus. Comparison of this nucleotide sequence to those of four previously published isolates revealed that one of the isolates, HM-175, which was obtained from Australia and passed three times in marmosets, had a 8.5-11% nucleotide variability compared to the remaining four isolates which were isolated from North American sources. This nucleotide variability does not result in amino acid differences with the exception of two of the North American isolates, which were derived from tissue culture passage. These isolates have been shown to contain regions of variability generated by nucleotide insertions and/or deletions, while the remaining three isolates, including the Australian isolate, demonstrate limited amino acid differences within the VP1 molecule. 相似文献
127.
Neurons in trigeminal and geniculate ganglia extend neurites that share contiguous target tissue fields in the fungiform papillae and taste buds of the mammalian tongue and thereby have principal roles in lingual somatosensation and gustation. Although functional differentiation of these neurons is central to formation of lingual sensory circuits, there is little known about electrophysiological properties of developing trigeminal and geniculate ganglia or the extrinsic factors that might regulate neural development. We used whole cell recordings from embryonic day 16 rat ganglia, maintained in culture as explants for 3-10 days with neurotrophin support to characterize basic properties of trigeminal and geniculate neurons over time in vitro and in comparison to each other. Each ganglion was cultured with the neurotrophin that supports maximal neuron survival and that would be encountered by growing neurites at highest concentration in target fields. Resting membrane potential and time constant did not alter over days in culture, whereas membrane resistance decreased and capacitance increased in association with small increases in trigeminal and geniculate soma size. Small gradual differences in action potential properties were observed for both ganglion types, including an increase in threshold current to elicit an action potential and a decrease in duration and increase in rise and fall slopes so that action potentials became shorter and sharper with time in culture. Using a period of 5-8 days in culture when neural properties are generally stable, we compared trigeminal and geniculate ganglia and revealed major differences between these embryonic ganglia in passive membrane and action potential characteristics. Geniculate neurons had lower resting membrane potential and higher input resistance and smaller, shorter, and sharper action potentials with lower thresholds than trigeminal neurons. Whereas all trigeminal neurons produced a single action potential at threshold depolarization, 35% of geniculate neurons fired repetitively. Furthermore, all trigeminal neurons produced TTX-resistant action potentials, but geniculate action potentials were abolished in the presence of low concentrations of TTX. Both trigeminal and geniculate neurons had inflections on the falling phase of the action potential that were reduced in the presence of various pharmacological blockers of calcium channel activation. Use of nifedipine, omega-conotoxin-MVIIA and GVIA, and omega-agatoxin-TK indicated that currents through L-, N-, and P/Q- type calcium channels participate in the action potential inflection in embryonic trigeminal and geniculate neurons. The data on passive membrane, action potential, and ion channel characteristics demonstrate clear differences between trigeminal and geniculate ganglion neurons at an embryonic stage when target tissues are innervated but receptor organs have not developed or are still immature. Therefore these electrophysiological distinctions between embryonic ganglia are present before neural activity from differentiated receptive fields can influence functional phenotype. 相似文献
128.
129.
Serodiagnosis of viral hepatitis A by a modified competitive binding radioimmunoassay for immunoglobulin M anti-hepatitis A virus. 总被引:3,自引:0,他引:3 下载免费PDF全文
D W Bradley H A Fields K A McCaustland J E Maynard R H Decker R Whittington L R Overby 《Journal of clinical microbiology》1979,9(1):120-127
A competitive binding radioimmunoassay (CBA) for antibody to hepatitis A virus (HAV) was evaluated and compared with a standard solid-phase radioimmunoassay for anti-HAV, CBA was found to be sensitive and specific for the detection of anti-HAV, as demonstrated by the 98% concordance of CBA and solid-phase radioimmunoassay test results. The standard CBA test was modified for the differential detection of acute (immunoglobulin M) and convalescent (immunoglobulin G) anti-HAV by incorporation of a step in which immunoglobulin G anti-HAV was preferentially absorbed with S. aureus cells (protein A). The modified CBA test was shown to be capable of differentiating between acute- and convalescent-phase sera. The modified CBAM test was able to detect immunoglobulin M anti-HAV up to approximately 4 weeks after the onset of illness. 相似文献
130.
A discrete stimulus (flashing light) was paired with cocaine (20 mg/kg) to induce conditioned locomotion. To identify brain regions activated during this response, Fos was measured with immunohistochemistry. Although paired subjects displayed robust conditioned locomotion, Fos was not increased in any limbic brain regions analyzed. In contrast, pairing of cocaine with generalized contextual cues (whole room) produced conditioned locomotion and Fos activation in the prelimbic portion of prefrontal cortex and the nucleus accumbens core. These results suggest that the pattern of neuronal activation during cocaine-conditioned activity differs depending on whether a discrete or contextual stimulus is used as a conditioned stimulus. The possibility that expectancy and frustrative nonreward contribute to Fos expression in rats conditioned to contextual cues is discussed. 相似文献