Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P30536 (PBS)
 9,886 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

An enzyme-linked immunosorbent assay (ELISA) for detecting antibody to Mycoplasma hyopneumoniae in porcine serum is described. The results are presented as an ELISA ratio, calculated by dividing the absorbance of the test sample by the mean absorbance of control negative sera. In known infected pigs, the ELISA ratio was highest when the serum concentration applied to the ELISA plate was diluted 1 in 20 in PBS - Tween. Mean ELISA ratios ranged from 1.2 +/- 0.3 for pigs without porcine enzootic pneumonia (PEP) lesions to 5.5 +/- 1.5 for pigs observed with a PEP lesion reacting positively with immunofluorescent histopathology. Pigs observed with typical PEP lesions at slaughter, but not confirmed by immunofluorescent histopathology had a mean ELISA ratio of 4.9 +/- 1.7. The ELISA was highly sensitive (95.6%) and specific (98.8%) when pig sera from commercial piggeries of known M hyopneumoniae infection status were assessed. No cross-reactivity with serum from a pig hyperimmunised with killed M flocculare was detected, and reactivity with serum from another pig hyperimmunised with killed M hyorhinis showed only weak cross-reactivity, which failed to reach the ELISA positive threshold (ELISA ratio 3) for M hyopneumoniae.
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PMID:Evaluation of an enzyme-linked immunosorbent assay for the detection of Mycoplasma hyopneumoniae antibody in porcine serum. 144 72

The DNA repair enzyme uracil-DNA glycosylase from Mycoplasma lactucae (831-C4) was purified 1,657-fold by using affinity chromatography and chromatofocusing techniques. The only substrate for the enzyme was DNA that contained uracil residues, and the Km of the enzyme was 1.05 +/- 0.12 microM for dUMP containing DNA. The product of the reaction was uracil, and it acted as a noncompetitive inhibitor of the uracil-DNA glycosylase with a Ki of 5.2 mM. The activity of the enzyme was insensitive to Mg2+, Mn2+, Zn2+, Ca2+, and Co2+ over the concentration range tested, and the activity was not inhibited by EDTA. The enzyme activity exhibited a biphasic response to monovalent cations and to polyamines. The enzyme had a pI of 6.4 and existed as a nonspherical monomeric protein with a molecular weight of 28,500 +/- 1,200. The uracil-DNA glycosylase from M. lactucae was inhibited by the uracil-DNA glycosylase inhibitor from bacteriophage PBS-2, but the amount of inhibitor required for 50% inhibition of the mycoplasmal enzyme was 2.2 and 8 times greater than that required to cause 50% inhibition of the uracil-DNA glycosylases from Escherichia coli and Bacillus subtilis, respectively. Previous studies have reported that some mollicutes lack uracil-DNA glycosylase activity, and the results of this study demonstrate that the uracil-DNA glycosylase from M. lactucae has a higher Km for uracil-containing DNA than those of the glycosylases of other procaryotic organisms. Thus, the low G + C content of the DNA from some mollicutes and the A.T-biased mutation pressure observed in these organisms may be related to their decreased capacity to remove uracil residues from DNA.
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PMID:A mollicute (mycoplasma) DNA repair enzyme: purification and characterization of uracil-DNA glycosylase. 234 31

A study was carried out to determine the efficacy of different adjuvants in enhancing antibody response to sonicated F-38 antigens. Goats were immunised against CCPP using antigens incorporated in Freund's incomplete adjuvant (IFA), saponin, aluminium hydroxide gel and buffered saline (PBS) respectively. Antibody responses were determined. The goats were challenged four months after immunisation to assess their immune status. Two of eight goats given antigen in PBS, six of 10 goats given antigen in aluminium hydroxide, seven of eight goats given antigen in IFA and all 10 goats given antigen in saponin withstood the challenge. Saponin and IFA were similar in their immune potentiation ability and were superior to aluminium hydroxide. As IFA has been considered unsuitable for use in food animals saponin may prove valuable in vaccination of goats against CCPP caused by mycoplasma strain F-38.
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PMID:Efficacy of different adjuvants to potentiate the immune response to mycoplasma strain F-38. 335 56

The effect of sonication on the viability of Mycoplasma salivarium and Mycoplasma orale was examined as a preliminary investigation on the quantitative studies on mycoplasmas in the dental plaques. Mycoplasma cell aggregations suspended in PBS or liquid medium were sonicated and the samples were removed at 5-second intervals for 20 seconds to check the viable counts. Inactivation of the mycoplasmas was almost directly proportional to the sonication time. M. orale was more sensitive to sonication than M. salivarium. Mycoplasmas were damaged more readily in PBS than in the liquid medium. Dental plaques suspended in PBS were also sonicated exactly as mentioned above. The number of mycoplasmas in the dental plaques (1 mg) ranged from 1.09 x 10(2) to 1.75 x 10(5) cfu. The desirable sonication time to prepare the homogeneous suspensions of dental plaques was concluded to be 5 seconds.
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PMID:A preliminary study on isolation and enumeration of mycoplasmas in dental plaques: the effect of sonication on viability of oral mycoplasmas. 694 63

The application of three photosensitive agents for disinfection of bovine semen was investigated. Bovine microbial pathogens suspended in tissue culture medium and/or PBS and also added to bovine semen were exposed to the photosensitive agents followed by irradiation. Hematoporphyrin, hematoporphyrin derivative and thiopyronine were effective against bovine herpes virus-1, bovine viral diarrhoea virus, Mycoplasma bovigenitalium, Mycoplasma canadense, and Ureaplasma diversum in culture media. In addition, thiopyronine was effective against Leptospira pomona. Similar treatments were not effective against Leptospira hardjo, Mycoplasma bovis, or Campylobacter fetus subsp. venerealis. When microorganisms were added to bovine semen, only bovine herpes virus-1 was controlled by the photosensitive agents used at concentrations which did not appear harmful to sperm cells.
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PMID:Studies on inactivation of pathogenic microorganisms in culture media and in bovine semen by photosensitive agents. 801 31

Mycoplasma salivarium cells were demonstrated to bind human IgG Fc fragments. The binding capacity was 78.8% enhanced by incubation of the cell suspension in PBS with 0.25% trypsin at 37 degrees C for 1 h, but tended to fall after incubation with higher concentrations of the enzyme, and was 95.5% lower after incubation of the suspension without trypsin. Fc fragments of IgG from rat, swine, sheep, rabbit, goat, cow and mouse also bound to the organism cells with increasing affinity in this order. The affinity of human IgG Fc fragments was almost comparable with those of sheep and rabbit. Antigen specific IgG from goat (specific for gamma-chain of human IgG, and mu-chain of human IgM) and rabbit (specific for whole molecules of goat IgG) bound to the cells. Binding of goat IgG Fc fragment was inhibited in the presence of antigen specific goat IgG (specific for gamma-chain of human IgG). These results suggest that M. salivarium cells bind IgG from a variety of animal species via the Fc fragment.
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PMID:Binding of Fc fragments of IgG from human and seven animal species to Mycoplasma salivarium cells. 829 54

Mycoplasma gallisepticum- or M. synoviae-challenged chickens were monitored with serological assays (serum plate agglutination, hemagglutination inhibition, and enzyme-linked immunosorbent assay) and polymerase chain reaction (PCR). The tracheal swabs from M. gallisepticum-challenged chickens received three different treatments (phosphate-buffered saline [PBS], Frey's broth, or 10 mM Tris-HCl/250 mM ethylenediaminetetraacetic acid/ 2.5% sodium dodecyl sulfate [STE]) prior to DNA purification. A nonphenolic method for DNA extraction was utilized. The best PCR results were obtained with PBS swab treatment. The nonphenolic method for DNA extraction was compared with a phenolic method in an experiment with tracheal swabs from M. synoviae-challenged chickens and commercial flocks. Both methods gave comparable results.
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PMID:Polymerase chain reaction optimization for Mycoplasma gallisepticum and M. synoviae diagnosis. 871 37

Actinobacillus pleuropneumoniae is the causative agent of porcine pleuropneumonia. The major adhesin of A. pleuropneumoniae has previously been identified as a lipopolysaccharide (LPS), and more recently, we demonstrated that high molecular mass LPS were involved in A. pleuropneumoniae adherence to porcine respiratory tract cells. We postulated that immunization with a LPS-based vaccine may confer a protective immunity. The high molecular mass O-polysaccharides obtained after acid hydrolysis and chromatographic separation were conjugated to bovine serum albumin (BSA) as a protein carrier. Groups of mice were injected twice with the following antigen preparations: whole-cell preparation, outer membrane preparation, O-polysaccharide-BSA conjugate, hydrolyzed LPS and phenol/water extracted LPS. A combination of different adjuvants was also used during these immunization procedures to induce a stronger immunological response to the polysaccharide antigen. Two weeks after the second injection, the mice were challenged intranasally with either homologous A. pleuropneumoniae serotype 1 strain or a serotype 5 strain. The highest survival rate, up to 80%, compared to the control groups (P < 0.05), was recorded when the mice were injected twice with 15 micrograms of carbohydrates of O-polysaccharide-BSA conjugate mixed with the saponin-derived adjuvant Quil A. Survival rates of between 60 and 70%, twice those observed in the control groups immunized with PBS, were recorded in mice injected with the O-polysaccharide-BSA conjugate mixed with other adjuvant preparations such as alhydrogel, peanut oil and Freund's incomplete adjuvant. However, the protection induced by the conjugate antigen preparation was serotype specific, because mice challenged with a serotype 5 strain were killed. Taken together, these results confirm the important role of A. pleuropneumoniae LPS in pathogenesis.
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PMID:Evaluation of protective efficacy of an Actinobacillus pleuropneumoniae serotype 1 lipopolysaccharide-protein conjugate in mice. 902 43

Indirect evidence suggests that innate immune mechanisms involving alveolar macrophages (AMs) are of major importance in antimycoplasmal defense. We compared the effects of AM depletion on intrapulmonary killing of Mycoplasma pulmonis during the early phase of infection in mycoplasma-resistant C57BL/6NCr (C57BL) and mycoplasma-susceptible C3H/HeNCr (C3H) mice. More than 80% of AMs were depleted in both strains of mice by intratracheal insufflation of liposome-encapsulated dichloromethylene bisphosphonate (L-Cl2MBP), compared to no significant AM depletion in either strain following insufflation of liposome-encapsulated phosphate-buffered saline (L-PBS), PBS alone, or no treatment. AM-depleted (L-Cl2MBP) and control (L-PBS) mice were infected intranasally with 10(5) CFU of M. pulmonis UAB CT, and their lungs were quantitatively cultured to assess intrapulmonary killing at 0, 8, 12, and 48 h postinfection. AM depletion exacerbated the infection in C57BL mice by reducing killing of the organism to a level comparable to that in C3H mice without AM depletion. In contrast, AM depletion did not alter killing in C3H mice. These results directly identify the AM as the main effector cell in early pulmonary antimycoplasmal defense and suggest that differences in mycoplasmal killing by AMs may explain the resistance of C57BL mice and the susceptibility of C3H mice to mycoplasmal infection.
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PMID:Depletion of alveolar macrophages exacerbates respiratory mycoplasmosis in mycoplasma-resistant C57BL mice but not mycoplasma-susceptible C3H mice. 916 64

The major adhesin of Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia, has been previously identified as the lipopolysaccharide (LPS). Experiments in our laboratory have shown that mice immunised with different A pleuropneumoniae serotype 1 LPS preparations were protected against a challenge with a virulent A pleuropneumoniae serotype 1 isolate. The purpose of the present study was to evaluate the protection of pigs against experimental A pleuropneumoniae infection following immunisation with two of these LPS preparations. Groups of five specific pathogen free (SPF) pigs were injected twice with one of the following antigen preparations: detoxified LPS, O-polysaccharide-BSA conjugate, a commercial bacterin, or PBS. Two weeks after the second injection, pigs were challenged intranasally with a virulent A pleuropneumoniae serotype 1 strain. Upon macroscopic examination, fibrino-haemorrhagic pleuropneumonia, compatible with A pleuropneumoniae infection, was observed in one to four pigs in each group. The more extensive lesions were present in control, unimmunised pigs and in animals vaccinated with the O-polysaccharide-BSA conjugate. The highest survival rate was recorded when the pigs had been immunised with detoxified LPS or the commercial bacterin. Taken together, our results suggest that a protection comparable with the one obtained with a commercial bacterin was observed when pigs were immunised with a single class of molecules, detoxified LPS. Most importantly, these results confirm the important role of A pleuropneumoniae LPS in protection against porcine pleuropneumonia. Finally, our results also support the idea that mice are not an appropriate model for the evaluation of porcine pleuropneumonia vaccines.
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PMID:Evaluation of the protective efficacy of Actinobacillus pleuropneumoniae serotype 1 detoxified lipopolysaccharides or O-polysaccharide-protein conjugate in pigs. 983 97


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