We isolated porcine cytomegaloviruses (PCMVs) from lung samples of fattening pigs collected in the slaughter houses of 4 prefectures of Japan. Seven isolates were obtained and used for a comparison ...of serological characteristics by ELISA. J1, the first field isolate in Japan, and B6 which was isolated in the UK were also used in the study. The serological relationships between the isolates were analysed by the method of Archetti and Horsfall. OF1 showed serological differences with Chiba2 and Hiroshima. Differences were also observed between Chiba2 and ChibaC, ChibaC and Kagawa. B6 showed differences with OF2, Chiba3, ChibaC and Hiroshima.
A monoclonal antibody (B9) was generated by using a rat malignant fibrous histiocytoma (MFH)-derived cloned cell line (MT-8) as the immunogen. Immunohistochemically, B9 reacted specifically with a ...cytoplasmic antigen of MT-8 cells. Furthermore, B9 immunolabeled another MFH-derived cloned cells (MT-9) and histiocytic sarcoma cells, as well as macrophages/histiocytes in normal and diseased tissues of rats. These findings suggest the presence of a common antigen recognized by B9 between MFH cells and macrophages/histiocytes. This suggests that MFH cells may express histiocytic nature.
Sandwich enzyme-liked immunosorbent assay (ELISA) was developed for the quantitative estimation of Clostridium perfringens enterotoxin (CPE) with monoclonal and polyclonal antibodies as capturing and ...detecting antibodies, respectively. The dose-dependent relationship between absorbance 405 nm and concentration of purified CPE was obtained over the range of 0.64-400 ng/ml. The sandwich ELISA was found to detect crude CPE in culture and CPE in 10% fecal extracts. This method is convenient, rapi
To develop a rapid and specific method to detect and/or identify enterohemorrhagic Escherichia coli O157:H7, two mouse monoclonal antibodies (MAbs) were prepared. Specificities of these two MAbs (1D9 ...and 3E8) were determined by flow cytometry method (FCM). MAbs 3E8 and 1D9 were found to react with E. coli O157:H7, Citrobacter freundii and Salmonella group N (O:30), but not with Escherichia hermannii. With a mixture containing strains of E. coli O157:H7 and E. coli O6:H1, two different peaks appeared in FCM with MAbs, whereas a single peak appeared with polyclonal rabbit antiserum. From these findings, FCM with MAb is suggested to be a rapid, specific, and useful method to detect and identify strains(s) of E. coli O157:H7 in food ingredients
A monoclonal antibody (MAb) reactive with 36 field isolates and 2 laboratory strains of feline calicivirus (FCV) was produced by immunizing mice with the mixture of FCVs. The MAb (4D7) reacted with ...FCVs in an enzyme-linked immunosorbent assay (ELISA), but had no neutralizing activity against the F4 strain of FCV. MAb 8G1, previously produced against the FCV F4 strain, also reacted in ELISA with all FCVs used in the present study. However, the epitopes recognized by 4D7 and 8G1 were different. Using these two MAbs and a polyclonal rabbit antibody, we attempted to develop a sandwich ELISA for detection of FCV antigen. The combination of 4D7 and the polyclonal rabbit IgG was most sensitive. Using this system, all the field isolates of FCV cultured in vitro were detected. However, among the 36 swab samples, from which FCV was isolated, 4 were negative.
We have previously undertaken preliminary characterization of twoRhipicephalus appendiculutusserine protease inhibitors (RAS-1 and -2) as anti-tick vaccine candidates. In this study, to clarify this ...hypothesis, we generated and further characterized recombinant RAS-1 and -2 (rRAS-1 and -2) and tested their potency as a cocktail anti-tick vaccine in cattle. RT-PCR analysis showed that RAS-1 and -2mRNA transcripts are expressed during all life cycle stages of ticks, independent of sex. As judged by SDS-PAGE rRAS-1 and -2 migrated as a molecular weight of around 64 and 60kDa protein, respectively, considering that the expression vector produced a recombinant protein fused with 18-22kDa TRX protein. RAS-1 and -2 were found not to be secreted into the bite site as determined by the reactivity of anti-tick saliva sera to rRAS-1 and -2, suggesting that both proteins are concealed antigens. Vaccination of cattle with a combination of rRAS-1 and -2 conferred significant protective immunity against ticks, resulting in 61.4% reduction in nymph engorgement rate, and in 28 and 43% increased mortality rate in adult female and male ticks, respectively. This is the first report on an anti-tick vaccine trial using a combination of two different serpins derived fromR. appendiculatus, and using cattle as a natural host.
The intercalation reaction of diclofenac sodium (DFS) with layered inorganic compounds, γ-titanium phosphate (γ-TiP), proton type titanium oxide (H-TiO2) and sodium type synthetic mica (Na-TSM), was ...examined on. The direct reaction of DFS in ethanol-water mixed solvent resulted in the large amount accommodation of DFS. The amount of intercalated DFS was the order of γ-TiP>H-TiO2>Na-TSM corresponding to the order of acidity. The intercalation using phospholiopids was also examined to assist the intercalation reaction. However, the amount of intercalated DFS was rather small in comparison with those in the direct reaction. DFS accommodated in γ-TiP dissolved into neutral and basic buffer solution stoichiometry while scarcely dissolved in the acidic solution. The mechanism of the intercalation and reverse dissolution was successfully accounted according to the ion-exchange mechanism between Na+ in DFS and H+ in γ-TiP. The dissolution from tablet of DFS/γ-TiP intercalation compound was examined by using a disintegrator. It was found that the dissolution rate appropriately controlled by mixing the disintegrator. The present results suggested the different possibilities in the clinical field to use layered inorganic compounds such as drug delivery system (DDS).
In bitches, the onset of pyometra, an infection of the uterus, characteristically occurs in the first half of the diestrous stage in the estrous cycle, in which the blood concentration of ...progesterone peaks and that of estradiol-17
β is lowest. To investigate the immunological mechanisms governing stage-specific onset of pyometra, peripheral blood mononuclear cells (PBMNCs) were collected from beagle bitches during different stages of the estrous cycle and examined using various immunological assays. When we examined the proliferative response of PBMNCs to PYO-252, that is a clone of
Escherichia coli isolated from the uterus of a dog afflicted with pyometra, the response of PBMNCs significantly decreased in the first half (day 10) of diestrus, but increased in proestrus/estrus. No significant differences were observed in the responses to concanavaline A between stages of the cycle. Throughout the estrous cycle, canine PBMNCs did not respond to lipopolysaccharide derived from
E. coli. The response of PBMNCs collected in anestrus to PYO-252 was significantly enhanced upon the addition of estradiol-17
β to the culture. In contrast, these responses were significantly suppressed in the presence of progesterone. Progesterone progenitor or metabolite molecules, which have a low affinity for the progesterone receptor, did not affect proliferative responses. Expression of gamma interferon (IFN
γ) in response to PYO-252 was also significantly enhanced by estradiol-17
β, but suppressed by progesterone. This evidence suggests that in the first half of the diestrous stage, suppressed activity of cellular immunity results from increasing progesterone concentration and minimal estrogen release. This marked decrease of immune resistance allows the expansion of
E. coli, which enter the uterine cavity through the loosened cervical canal during estrus, leading to pyometra onset.
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