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)

Two-hundred sputum specimens from tuberculosis patients were examined for viable or non-viable mycobacteria by a combination of fluorescein diacetate ethidium bromide (FDA/EB) staining, Ziehl-Neelsen staining, and the use of cultures in 3% Ogawa egg medium. The sputum specimens were treated with 3% NaOH for 10 min and washed in PBS. The bacteria was then harvested by centrifugation at 6,000 rpm for 5 min. Each sample was subjected to FDA/EB staining, Ziehl-Neelsen staining and cultures to compare the staining -method results and the results of colony formation. Ziehl-Neelsen staining method revealed acid-fast bacteria in the specimens, distributed from Gaffky 1 to Gaffky 8. The number of FDA-positive specimens and culture-positive specimens were identical in all Gaffky grades, suggesting that the FDA staining method well reflected the presence of viable mycobacteria in the specimens. We concluded that FDA staining is a valuable method to detect viable mycobacteria in sputum specimens on the first day of examination. It is therefore advantageous for doctors and patients to be immediately informed of culture results rather than waiting for several weeks.
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PMID:[Application of FDA/EB staining for the detection of viable or non-viable mycobacteria in clinical specimens]. 137 66

In this report we are able to show that intravenous (i.v.) application (day 0) of a nonapeptide (residues 26-34) from the human C1q A-chain (designated peptide A-C1q) prior to intradermal (i.d.) administration of chicken type II collagen (CII) in arthritis-susceptible DBA/1 mice (H2q), leads to abrogation of polymorphonuclear neutrophil (PMN) invasion into the joints. This nonapeptide exhibits epitope characteristics and high homology to residues 137-147 of CB11 (a cyanogen bromide fragment of chicken CII, known to contain both arthritis inducing and suppressing determinants). Arthritis index was lowest in animals pretreated i.v. with CII (as internal control), though animals pretreated i.v. with peptide K (residues 137-147 with an additional glycine residue from CB11) or peptide A-C1q exhibited comparative arthritic indices. Only in the arthritis-positive control group (day 0: PBS i.v.) did i.d. application of CII lead to invasion of PMN into the synovial layer and the joint space. Analysis of antibody (Ab) responses at day 48 after i.v. immunization (day 0) and CII challenge (day 7) revealed IgE-Abs to native CII and also to native C1q. IgG titers to CII were highest in animals pretreated with peptide A-C1q. Abs from this group, exhibiting activity to peptide A-C1q (immunizing antigen), were of mainly IgG1 and IgG3 isotypes. Evaluation of the immune response following i.v. application of peptide A-C1q or CII, prior to i.d. CII administration, in DBA/1 mice, revealed IgM responses to peptide A-C1q and peptide K, but not to CII. Intravenous application of peptide A-C1q led to generation of IgG3-Abs reacting only with peptide A-C1q and peptide K, but not with native CII. Additionally, i.v. application of peptide A-C1q elicited IgG responses to a pentapeptide, resembling amino acid residues 26-30 (K-G-E-Q-G) of the C1q A-chain. This five residue antigenic determinant is present in peptide K, in chicken and human CII as well as in human C1q. No specific IgE response to any of the antigens tested could be detected. Since a peptide from the C1q A-chain is both capable of eliciting immune responses and modulating CII-induced arthritis in mice, we postulate that the collagen-like complement component C1q is involved in the development of CII-induced inflammatory arthritic lesions, and may represent, in vivo, the early antigen responsible for inducing anticollagen antibodies prior to CII in hyaline cartilage becoming available as antigen.
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PMID:Modulation of type II collagen-induced arthritis in DBA/1 mice by intravenous application of a peptide from the C1q-A chain. 139 37

Acid-fast staining is broadly used for the detection of mycobacteria in smears or pathological preparations, but it does not distinguish viable and non-viable bacteria. Enzymatic hydrolysis of fluorescein diacetate (FDA) was reported as a tool to detect viable mammalian cells or protozoa. We have applied this method to mycobacteria samples to detect viable or non-viable bacteria in the smear. Mycobacterium bovis BCG (Tokyo), 20 mg/ml suspension of standard vaccine, was used as the sample of viable bacteria. A part of the same suspension was heated at 100 degrees C for 30 min and used as the sample of non-viable bacteria. Three samples, viable, non-viable, and 1:1 mixture of the two, were stained with acid-fast staining and FDA-EB (ethidium bromide) mixture, respectively. For FDA/EB staining, stock solution of FDA (5 mg/ml acetone) was diluted 1:50 in PBS and 25 microliters of FDA and 25 microliters of EB (20 micrograms/ml PBS) were mixed with 50 microliters of bacterial suspension. Preparations were observed with either light or fluorescein microscope. Living bacteria were stained in yellow green in FDA/EB staining while non-viable bacteria were red. Mixed sample of live and dead bacilli showed differential staining with green or red in FDA/EB staining, but no difference was shown in acid-fast staining between viable and non-viable bacteria.
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PMID:[Detection of viable or non-viable mycobacteria by hydrolysis of fluorogenic substrate]. 169 50

The asynchronous development of Eimeria tenella in orally infected chickens makes it possible to purify second generation merozoites (meros) and shizonts from a single mucosal homogenate. After centrifugation in 30% Percoll in phosphate-buffered saline (Percoll-PBS), debris, villi, and schizonts float, whereas meros and erythrocytes are pelleted. Erythrocytes are lysed by a mild hypotonic shock; meros are filtered through a cotton wool plug and collected by centrifugation. The 30% Percoll-PBS supernatant fraction is diluted to 25% Percoll-PBS and centrifuged to sediment mature schizonts. By repeated slow-speed centrifugation, schizonts are separated from nuclei and small-sized debris. In less than 3 hr, 8.8 +/- 2.3 x 10(8) meros and 7.2 +/- 3.9 x 10(6) schizonts are collected from 10 infected chickens. Contamination with host material is 2% for meros but variable for schizonts. For the assessment of cell viability, ethidium bromide (EB) and acridine orange (AO) have been used as markers for dead and living cells, respectively, in a single step method. More than 95% of the schizonts and meros accumulate AO and no EB, whereas lysed erythrocytes and all cells hosting a schizont are permeable to EB. After incubation of meros and schizonts in synthetic media with [5,6- 3H]uracil, label accumulates in the perchloric acid-soluble and -insoluble fractions, indicating transport, salvage, and incorporation of the pyrimidine precursor in nucleic acids. If stored on ice, meros and schizonts retain metabolic activity for at least 5 hr, but metabolism declines rapidly during incubation at 41 C.
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PMID:Simultaneous purification of merozoites and schizonts of Eimeria tenella (Apicomplexa) by Percoll flotation and assessment of cell viability with a double fluorescent dye assay. 177 4

Three experiments were performed to evaluate the inflammatory response, the antibody response and protection from experimental challenge of various Actinobacillus pleuropneumoniae serotype 5 (Ap5) vaccines in swine. In the first experiment, subcutaneous injections of either a water-in-oil (W/O) emulsion or Freund's complete adjuvant (FCA) caused lesions at the site of injection, while intraperitoneal injection of the W/O emulsion caused no lesions. In the second experiment, intraperitoneal (IP) injection of a W/O emulsion containing unwashed Ap5 cells (6-h culture) and/or supernates from a 24-h culture resulted in severe peritoneal lesions, while W/O emulsion containing PBS-washed Ap5 cells resulted in minimal peritoneal lesions. Ap5 alone or W/O alone failed to cause peritoneal lesions. The third experiment compared the antibody response and protection from challenge of pigs immunized with either 6-h PBS-washed Ap5 cells emulsified in oil - IP, 6-hour Ap5 cells adjuvanted with dimethyl diodacyl ammonium bromide - IP, Ap5 antigen alone - IP, a commercial vaccine - subcutaneously or saline - IP. All groups, except the saline-treated group, responded with high antibody titers to Ap5 2 weeks following vaccination; however, titers from the W/O plus antigen group were significantly higher than the three other groups (P less than 0.05). Following intranasal challenge with Ap5, all animals responded with increased antibody titers. All pigs were euthanized 10 days after challenge and evaluated for pneumonia and the lungs cultured for bacteria. The lungs of all pigs, excepting the W/O plus antigen group, contained pneumonic lesions and A. pleuropneumoniae was cultured from these lesions. These results, along with results from other groups, suggest that intraperitoneal immunization using oil-adjuvanted vaccine may be an effective method for protecting pigs from pneumonia due to A. pleuropneumoniae. Its efficacy may be due to stimulation of local respiratory mucosal immunity.
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PMID:Comparison of protective immunity and inflammatory responses of pigs following immunization with different Actinobacillus pleuropneumoniae preparations with and without adjuvants. 281 78

An ELISA was developed to measure human IgG antibody to the native polysaccharide antigen of GBS serotype Ia. Because the polysaccharide binds poorly to polyvinyl chloride, its adherence was enhanced by activation with cyanogen bromide and coupling to HSA in a molar ratio of polysaccharide to albumin of 1:4.5. There was minimal loss of sialic acid during coupling, and the coupled antigen showed identity with uncoupled native antigen by Ouchterlony analysis. OD values obtained by ELISA showed a log-linear relation to concentration of specific antibody in whole and affinity-chromatographed human sera measured by quantitative precipitation over a range of 0.25 to 3.5 microgram/ml. In replicate ELISA experiments using serially diluted human serum, dilutions with antibody content as low as 0.016 microgram/ml could be reliably differentiated from PBS or agammaglobulinemic serum. The concentration of antibody in 98 selected human sera measured by ELISA correlated well (r = 0.89, p less than 0.001) with results obtained by indirect IF assay. This quantitative ELISA for GBS antibody is rapid, convenient, economical, and suitable for routine use.
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PMID:An enzyme-linked immunosorbent assay (ELISA) for human IgG antibody to the type Ia polysaccharide of group B streptococcus. 705 Feb 70

Treatment of human lymphocytes with hydrogen peroxide (10 microM, 30 min, 37 degrees C in PBS) or with 1 cGy X-rays evoked about a 30% decrease in the frequency of micronuclei upon subsequent X-irradiation (1.5 Gy). In addition to a lower micronuclei frequency, we also found an increase in the sedimentation distance of the nucleoids, when measured 90 min (duration of the isolation procedure carried out at 4 degrees C) after the adaptive dose (hydrogen peroxide or X-rays) and preceding the challenge dose. To test whether Ca2+ is involved in the induction of the adaptive response pathway, we treated cells with the calcium chelator, EGTA. When EGTA was given at the same time as the adaptive dose, it prevented the development of the adaptive response. In addition, the calcium antagonist, TMB-8, also prevented the development of the adaptive response as it prevented the reduction of both micronuclei and increased nucleoid sedimentation. Cellular treatment with TMB-8 increased the free [Ca2+] by 40%, when given together with hydrogen peroxide. The faster sedimenting nucleoids from adapted cells were also examined by ethidium bromide titration; there was no indication of any change in supercoil density or loop size. Psi-tectorigenin, an inhibitor of phosphatidylinositol turnover, did not modify the adaptive response, indicating that inositol (1,4,5)-trisphosphate is not involved in the induction of the adaptive response, but free Ca2+ ions are.
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PMID:Calcium antagonist, TMB-8, prevents the induction of adaptive response by hydrogen peroxide or X-rays in human lymphocytes. 802 16

The nucleus of the solitary tract (NTS) was systematically explored in the alpha-chloralose-anesthetized rat for sites that elicited changes in mean arterial pressure (MAP) and heart rate (HR) during microinjections (20 nl) of phosphate-buffered saline (PBS; pH 7.2-7.4) or NaCl solutions containing various concentrations of NaCl (104-326 mM). Decreases in MAP (range 7-83 mmHg) and HR (range 10-70 bpm) were consistently elicited from sites in the caudal medial and commissural subnuclei of NTS. Microinjection of PBS or NaCl into other NTS subnuclei or area postrema did not elicit cardiovascular responses. Microinjection of LiCl in PBS elicited cardiovascular responses that were significantly smaller than those elicited by microinjection of NaCl in PBS at the same NTS site. Injections of either a hyperosmotic (400 mOsm/kg) or a hyposmotic (204 mOsm/kg) solution of mannitol into NaCl-sensitive sites did not elicit cardiovascular responses. Finally, most of the sites in NTS that elicited cardiovascular responses during microinjection of glutamate (1 M) did not respond to microinjections of PBS. Administration of atropine methyl bromide had no effect on the magnitude of the depressor response to injections of PBS into NTS, but significantly attenuated (32%) the HR response. Subsequent administration of the ganglionic blockers hexamethonium bromide or arfonad abolished both the depressor and bradycardic responses. These data suggest that within a restricted region of the caudal NTS there exists a pool of neurons sensitive to changes in extracellular Na+ concentrations that, when activated by the sodium, elicit vasodepressor responses as a result of sympathoinhibition and bradycardia as a result of vagal excitation and sympathoinhibition.
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PMID:Cardiovascular effects of NaCl microinjections into the nucleus of the solitary tract. 805 35

We have recently demonstrated the presence of phospholipase A2 (PLA2) activity in cells from bovine retinal pigment epithelium (RPE) [Jacob et al. (1996) J. Biol. Chem. 271, 19209-19218]. We report here our results on the characterization of this RPE-PLA2 activity. We show that RPE probably contains two types of PLA2 enzyme, as indicated by the results obtained with different PLA2-active fractions eluted from cation-exchange columns and treated with Ca2+/EGTA, dithiothreitol, p-bromophenacyl bromide or heat. These results, in addition to those from PLA2 assays using different substrates, also suggest that RPE-PLA2 enzymes are different from the well-known secretory, cytoplasmic and Ca2+-independent forms. Sequential extraction of RPE with (1) isotonic, (2) hypertonic and (3) detergent-containing PBS argues for the presence of weakly membrane-associated enzymes. Control experiments using 'back and forth' TLC allowed us to discriminate between PLA2 and phospholipase C/diacylglycerol lipase activity and confirmed that, in our assay conditions, the release of fatty acids was indeed due to PLA2 enzymes. These results, together with those obtained by treating RPE homogenates with H2SO4, guanosine 5'-[gamma-thio]triphosphate, ATP and different protease inhibitors, permitted us to make the first characterization of these RPE-PLA2 enzymes. We conclude that RPE contains novel types of PLA2 that are different from the secretory, cytoplasmic and Ca2+-independent forms.
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PMID:Bovine retinal pigment epithelium contains novel types of phospholipase A2. 935 16

A study has been conducted in which HeLa cells are incubated with hematoporphyrin derivative (HpD) for 1 h (1 microgram/ml of HpD in PBS) to compare the use of continuous wave (CW) and pulsed laser (10 Hz repetition rate and 7-9 ns pulse width) light for photodynamic therapy. Cytotoxic effects on the cells are evaluated by the 3-(4,5-dimethyl-2-thiazolyl)-2-5-diphenyl-2H-tetrazolium bromide (MTT) assay and the fluorescein diacetate (FDA)/propidium iodide (PI) stain method using a flow cytometer. The type of cell death is estimated by analysis of the DNA content and observation of the nuclear morphology. The cytotoxicity ratio of cells irradiated by pulsed laser light is estimated to be lower than that for CW laser light. The viability of cells that received pulsed laser light gradually decreases, whereas no significant changes are found in the cells irradiated with CW laser light with the elapse of post-irradiation time. The type of cell death differs between the pulsed and CW laser light irradiations. These findings suggest that the cytotoxic efficacy of the excitation light source is displayed by the difference in the type of cell death, namely apoptosis or necrosis.
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PMID:Comparison of phototoxicity mechanism between pulsed and continuous wave irradiation in photodynamic therapy. 1067 29


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