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41.
Development of a radioimmunoassay for Escherichia coli heat-stable enterotoxin: comparison with the suckling mouse bioassay. 总被引:11,自引:9,他引:11
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Escherichia coli strains which produce heat-stable enterotoxin (ST) are usually identified by demonstrating the production of ST. At present, ST can be detected only by bioassay methods. Recently, we purified E. coli ST, which enabled us to develop a radioimmunoassay for ST. Radioiodination of ST was performed by the lactoperoxidase method, which resulted in a high specific activity and retained the biological activity of St. Anti-ST antisera were raised in goats by injecting the goats with pure ST coupled to bovine immunoglobin G. Antibody titers ranged from 1:8,000 to 1:40,000. Using these reagents, we examined assay conditions thoroughly and found that a 14- to 18-h incubation at 4 degrees C in sodium acetate buffer with an ionic strength of 120 mM (pH 6.2) gave maximal sensitivity and reproducibility. Free ST was separated from antibody-bound ST by dextran-coated charcoal. This radioimmunoassay accurately and reproducibly measured ST in the range from 50 to 500 pg of ST per tube and could quantitate ST accurately in complex bacteriological media. This assay was specific for STa, measured human and porcine STa equally well, and did not cross-react with STb, with several other enterotoxins, or with various gastrointestinal peptides. Intact disulfide bridges in the ST molecule were required for immunoreactive activity. 相似文献
42.
Inhibition of Bacteroides fragilis on blood agar plates and reversal of inhibition by added hemin. 总被引:2,自引:0,他引:2
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T D Wilkins S L Chalgren F Jimenez-Ulate C R Drake Jr J L Johnson 《Journal of clinical microbiology》1976,3(3):359-363
Bacteroides fragilis strains formed much smaller colonies on most types of blood agar plates than they did on the same media without blood. Blood inhibited strains of B. fragilis subsp. distasonis the most and B. fragilis subsp. fragilis the least. The inhibition could be eliminated by adding hemin to the blood agar. The inhibitory component of the blood was inside the erythrocytes and appeared to be the hemin-free globin of hemoglobin. 相似文献
43.
Comparison of chlorhexidine and tincture of iodine for skin antisepsis in preparation for blood sample collection
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Rates of contamination of blood cultures obtained when skin was prepared with iodine tincture versus chlorhexidine were compared. For iodine tincture, the contamination rate was 2.7%; for chlorhexidine, it was 3.1%. The 0.41% difference is not statistically significant. Chlorhexidine has comparable effectiveness and is safer, cheaper, and preferred by staff, so it is an alternative to iodine tincture. 相似文献
44.
Highly immunogenic and protective recombinant vaccine candidate expressed in transgenic plants 总被引:5,自引:0,他引:5
Chargelegue D Drake PM Obregon P Prada A Fairweather N Ma JK 《Infection and immunity》2005,73(9):5915-5922
Vaccine development has been hampered by difficulties in developing new and safe adjuvants, so alternative technologies that offer new avenues forward are urgently needed. The goal of this study was to express a monoclonal recombinant immune complex in a transgenic plant. A recombinant protein consisting of a tetanus toxin C fragment-specific monoclonal antibody fused with the tetanus toxin C fragment was designed and expressed. Immune complex formation occurred between individual fusion proteins to form immune complex-like aggregates that bound C1q and FcgammaRIIa receptor and could be targeted to antigen-presenting cells. Unlike antigen alone, the recombinant immune fusion complexes were highly immunogenic in mice and did not require coadministration of an adjuvant (when injected subcutaneously). Indeed, these complexes elicited antibody titers that were more than 10,000 times higher than those observed in animals immunized with the antigen alone. Furthermore, animals immunized with only 1 mug of recombinant immune complex without adjuvant were fully protected against lethal challenge. This the first report on the use of a genetic fusion between antigen and antibody to ensure an optimal expression ratio between the two moieties and to obtain fully functional recombinant immune complexes as a new vaccine model. 相似文献
45.
Evidence that amyloid beta-peptide-induced lipid peroxidation and its sequelae in Alzheimer's disease brain contribute to neuronal death 总被引:22,自引:0,他引:22
Amyloid beta-peptide [Abeta(1-42)] is central to the pathogenesis of Alzheimer's disease (AD), and the AD brain is under intense oxidative stress, including membrane lipid peroxidation. Abeta(1-42) causes oxidative stress in and neurotoxicity to neurons in mechanisms that are inhibited by Vitamin E and involve the single methionine residue of this peptide. In particular, Abeta induces lipid peroxidation in ways that are inhibited by free radical antioxidants. Two reactive products of lipid peroxidation are the alkenals, 4-hydroxynonenal (HNE) and 2-propenal (acrolein). These alkenals covalently bind to synaptosomal protein cysteine, histidine, and lysine residues by Michael addition to change protein conformation and function. HNE or acrolein binding to proteins introduces a carbonyl to the protein, making the protein oxidatively modified as a consequence of lipid peroxidation. Immunoprecipitation of proteins from AD and control brain, obtained no longer than 4h PMI, showed selective proteins are oxidatively modified in the AD brain. Creatine kinase (CK) and beta-actin have increased carbonyl groups, and Glt-1, a glutamate transporter, has increased binding of HNE in AD. Abeta(1-42) addition to synaptosomes also results in HNE binding to Glt-1, thereby coupling increased Abeta(1-42) in AD brain to increased lipid peroxidation and its sequelae and possibly explaining the mechanism of glutamate transport inhibition known in AD brain. Abeta also inhibits CK. Implications of these findings relate to decreased energy utilization, altered assembly of cytoskeletal proteins, and increased excitotoxicity to neurons by glutamate, all reported for AD. The epsilon-4 allele of the lipid carrier protein apolipoprotein E (APOE) allele is a risk factor for AD. Synaptosomes from APOE knock-out mice are more vulnerable to Abeta-induced oxidative stress (protein oxidation, lipid peroxidation, and ROS generation) than are those from wild-type mice. Further, synaptosomes from allele-specific APOE knock-in mice have tiered vulnerability to Abeta(1-42)-induced oxidative stress, with APOE4 more vulnerable to Abeta(1-42) than are those from APOE2 or APOE3 mice. These results are consistent with the notion of a coupling of the oxidative environment in AD brain and increased risk of developing this disorder. Taken together, the findings from in-vitro studies of lipid peroxidation induced by Abeta(1-42) and postmortem studies of lipid peroxidation (and its sequelae) in AD brain may help explain the APOE allele-related risk for AD, some of the functional and structural alterations in AD brain, and strongly support a causative role of Abeta(1-42)-induced oxidative stress in AD neurodegeneration. 相似文献
46.
47.
P S Bass A F Drake Y Wang J H Thomas D R Davies 《Laboratory investigation; a journal of technical methods and pathology》1990,62(2):185-188
The molecular charge on bovine serum albumin (BSA) was modified by substituting carboxyl groups on the protein with ethylenediamine, thereby producing a highly cationic derivative with a pI of 9.3 to 9.5. Gel-filtration studies showed that the molecular weight of BSA was not significantly altered after cationization. When the cationized BSA was administered to rabbits using a chronic serum sickness schedule of injections, the animals developed a membranous glomerulopathy similar to the human disease, except that approximately one-third of the animals also showed focal and segmental endocapillary proliferation. Comparison of the circular dichroism spectra of native and cationized BSA showed that the substitution of the carboxyl groups resulted in a 50% reduction in the alpha-helical content of the native molecule. This conformational change should be considered as a possible determinant of the different immune response and immunopathology associated with the cationized molecule compared with native BSA. 相似文献
48.
R L Drake 《The Anatomical record》1988,220(3):300-304
Insulin treatment of diabetic rats fed a high-carbohydrate, fat-free diet produces a dramatic accumulation of hepatic lipids. However, this increase in hepatic lipids may only be a response to injections of exceptionally high doses of insulin. This study addresses this possibility. Alloxan-diabetic rats, fed a high-carbohydrate, fat-free diet, were given insulin every 12 h for 60 h at the following dosages: 1/2 unit each, 1 unit each, 2 units each, and 4 units each of regular and NPH insulins. At the end of the treatment period, liver samples were collected and used for morphological and biochemical analyses. Histologic examination revealed hepatic lipid accumulations at all insulin doses; the amount of lipid increased until maximal levels were reached at an insulin dosage of 1 + 1, which was maintained at doses of 2 + 2 and 4 + 4. Thus, hepatic lipid accumulation occurs regardless of the dosage of insulin administered to the diabetic animal. It is not simply an abnormal cellular response to excessive hormone levels. Similarly, the activity of the hepatic lipogenic enzyme, malic enzyme, increased at initial insulin dosages and reached maximal levels at 2 + 2. However, in contrast to lipid accumulation, enzyme activity decreased at the final insulin dosage of 4 + 4. Thus, there appears to be a direct relationship between increasing insulin levels and malic enzyme activity until an optimal insulin concentration is reached. After this point, excessive insulin levels do inhibit malic enzyme activity. 相似文献
49.
From 1990 through 1994, we fortuitously isolated Histoplasma capsulatum from six patients with AIDS whose specimens of blood were processed by the BACTEC system using Middlebrook broth selective for acid-fast bacilli (13A medium). Growth indices became positive after an average of 17 days of incubation (range, 11 to 20 days). No acid-fast bacilli were seen, but small budding yeasts characteristic of H. capsulatum were present. 相似文献
50.