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Query: UNIPROT:P30536 (PBS)
 9,886 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The accuracy with which areas of bronchial mucous gland on histological sections may be determined has been investigated with respect to inherent statistical variability in the point counting procedure used, and with respect to variations between the histological stains and observers employed. Very good argeement was obtained between areas determined by point counting and by planimetry. It was shown empirically that for sections of well-defined gland-like structures the accuracy (coefficient of area) of area determination was inversely proportional to the 3/4 power of the mean point count. The constant of proportionality depended on the structure's shape and on the point geometry of the grid used. Using the relationship the count needed to achieve a required accuracy of area determination could be established. It was shown that in general a smaller count was required for a given accuracy than would be the case using randomly dispersed tissue. On histological sections the accuracy of area determination is also dependent upon gland boundary definition, and our experiments showed that a recticulin Alcian Blue stain best defined mucous gland acini, and that a PBS stain gave the most accurate results for whole gland. These experiments also showed that there were small but significant differences in mean areas determined by different observers. The observers were shown to be equally consistent in their judgment of what constituted whole mucous gland and acini, but since they differed over mean counts it is recommended that studies should be designed to use either a single observer or to assign observers randomly among groups. With these considerations point counting can be made an accurate method of area determination for these or similar tissues.
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PMID:A comparative study of the accuracy of area assessment by point counting for bronchial mucous glands. 72 8

Purified and desialylated glycoprotein M from human O erythrocytes precipitates with B and H specific lectin from Evonymous europaeus seeds, both in PBS and 0.2% Triton X-100. Desialylated, N-terminal fragment (MT-1) obtained by trypsin digestion of M glycoprotein does not precipitate with Evonymous lectin but inhibits precipitation.
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PMID:Localization and immunochemical characterization of the lectin Evonymous europaeus receptor site on the glycoprotein from human O erythrocytes. 74 64

Some basic problems in immunofluorescence were reexamined, limiting the object to kidneys. As to thickness of sections, 4 mum was demonstrated to be the most reasonable also to kidney specimens. Treatment of PBS for a short time before staining was recommended not only for cleanup effect but also for the possibility to disclose covered antigenic determinants. It was also established that fluorescent antibody with adequate F/P molar ratio, as well as with strict specificity, should be used, in order to prevent "false positives".
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PMID:The application of immunofluorescence to the study of renal disease. I. Influences of various conditions of specimens and conjugates on the fluorescent patterns of kidney. 77 46

A new model system for quantitation of immunofluorescence on single cells is described using poly-L-lysine (PLL) coated polyacrylic plastic beads of approximately cell sizes as carriers for protein antigen. By increasing PLL concentration on the beads increased amounts of 125I labeled antigen were attached to the particles. Excess binding sites of PLL could be completely blocked by unrelated proteins. After staining with FITC-conjugated antibodies and quantitative fluorescence measurements of individual beads using a microspectrofluorimeter, strong correlations were found between antibody and antigen concentration on the beads. Neither repeated washings with PBS nor storage of the beads for two months caused detectable shedding of antigen-antibody complexes. There was a strong linear correlation between fluorescence intensity and the volume of beads, but the correlation between surface area and fluorescence was nonlinear. The described procedure was shown to be a simple method for quantitative and stable coating of particles with proteins. It can be applied as a useful model system for quantitative immunofluorescence studies on intracellular antigens.
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PMID:Insolubilization of protein antigens on polyacrylic plastic beads using poly-L-lysine. 79 53

After mutagenesis with nitrosoguanidine, germination mutants of Bacillus subtilis 168 were selected by killing, with heat, spores that germinated at 42 C and collecting survivors at 30 C. The germination properties of nine mutants variously affected in amino acid biosynthesis and sugar utilization were studied in detail. They were divided into two groups: (i) Ger-ALA mutants, failed to germinate in 10 mM L-alanine but germinated in complex media (some of these mutants were temperature sensitive); (ii) Ger-PAB mutants, germinated poorly, even in complex media, suggesting that they were blocked in important germination functions. All the mutants failed to germinate in L-alpha-amino-n-butyrate or L-valine (including temperature-sensitive mutants only at the restrictive temperature) showing that there is a step necessary for germination affected by all three acids. The mutants had normal growth rates, indicating that the defective gene products were specific for germination functions. These defects were not identified. Eight of the mutants were mapped by transduction with phage PBS-1. The recombinants were scored either by observations, by microscopy of phase darkening of the spores, or by a plate test involving the reduction of tetrazolium by heated colonies of spores. Five of the mutations, of at least three phenotypes, were between thr-5 and cysB3 away from all the sporulation markers that have been previously mapped. A linked ald (alanine dehydrogenase) locus was on the other side of thr-5. The other Ger markers were located in at least two additional positions. Auxotrophic strains that were used for mapping germinated normally, but germination of the Ger mutants differed slightly in different genetic backgrounds.
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PMID:Isolation, characterization, and mapping of Bacillus subtilis 168 germination mutants. 80 83

Sheep red cells, sensitized with 19S fraction of antiserum and subsequently treated with mouse serum as the source of complement (EAC), interact with human diploid fibroblasts (WI-38 cells) and form "Rosettes". Under a scanning electron microscope, EAC have not attached directly to the cell surface of fibroblasts, but to the fine processes or microvilli of the latter, as if there were fine bridges between EAC and the surface of fibroblasts. On the other hand, the attachment of sheep red cells washed in PBS (E) or sensitized with 19S fraction of antiserum (EA) to WI-38 cells was not observed. The pretreatment of WI-38 cells with mouse serum did not inhibit the interaction of WI-38 cells and EAC. No phagocytosis of EAC by WI-38 cells was observed in the 2 hrs incubation of both cells. From these results it is suspected that the interaction of WI-38 cells and EAC is immune adherence, and that WI-38 cells have the receptor site for complement, especially for C3, on the surface of cell membrane.
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PMID:The recetor sites for complement (C3) on human diploid fibroblasts. 80 27

The replication order of markers was studied in Bacillus subtilis strains bearing the trpE26 mutation by the use of the density transfer technique. An important difference in this order was observed in comparison with that of strain 168 T-. All markers tested of a chromosome segment extending from trpD to ilvA replicated early, after purB6 and before thr-5. Two markers flanking this region, trpE8 and citK7, replicated late as usual. The results suggested that this segment was shifted in trpE26 strains to a region closer to the origin of replication. PBS-1-mediated transduction crosses corroborated this hypothesis and revealed the position of the translocated segment. (i) Linkage was demonstrated for markers in the segment (hisH2, tryA1, met B3, ilvA2) to thr-5 and ald; (ii) aroB2 and citK7 were found to be linked; and (iii) linkage of cysB3 to thr-5 was lost in trpE26 strains. These findings made it possible to account for the characteristics of the trpE26 mutation and to propose a model explaining the fact that all trpE26+ transformants or transductants are merodiploid. The model calls for fusion of two genetic elements: two independent chromosomes, or two arms of a replicating structure. The resulting chromosome bears a long tandem duplication. Most of the features of this system of merodiploid formation can be interpreted by use of this model: the segregation pattern of the diploids, the stabilization of the unstable clones, and the length of the duplicated region. A relatively stable diploid strain was also studied by the density transfer technique. The data show that it remained diploid for the region corresponding to the translocated segment and are in agreement with the structure predicted by the model.
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PMID:Evidence for the Translocation of a Chromosome Sement in Bacillus subtilis Strains Carrying the trpE26 Mutation. 80 60

We have isolated a new mutant of Bacillus subtilis temperature sensitive in DNA replication; its properties are those of an initiation mutant. When liquid cultures are shifted to 48 degrees DNA replication is the first macromolecular synthesis that stops, but only after synthesis of the amount of DNA predicted for the completion of one replication round. When spores of the mutant are germinated and shifted to 48 degrees at subsequent times, one round of DNA replication is observed only when the shift occurs between 60 and 100 min; earlier shifts do not allow replication to start, later shifts allow more than one replication. The DNA replicated after a shift to high temperature is enriched in markers close to the terminus. The reinitiation of DNA replication stopped by the high temperature, takes place following a shift to a permissive temperature only if protein synthesis is allowed. Examination of DNA replication following toluene treatment shows that the elongation of DNA chains is not affected at the non-permissive temperature. This mutant is shown by PBS-1 mapping to correspond to a new gene denominated dna P, which is located between the thy A and fur A genes and is distinct from all the mapped dna and rec genes of Bacillus subtilis. The mutation confers to the cells also a deficiency in the ability to be transformed, to be transfected with SPP1 phage DNA, and to survive treatment with methyl-methane sulfonate. These deficiencies, observed at the permissive temperature, are no more temperature dependent than in the parental strain. The ability to perform homologous and heterologous transduction with PBS-1 phage and the sensitivity to ultraviolet radiation or mitomycin C are normal.
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PMID:A new mutant of Bacillus subtilis altered in the initiation of chromosome replication. 81 Jun 58

Two inhibitors of replicative deoxyribonucleic acid (DNA) synthesis, nalidixic acid (NAL) and 6-(p-hydroxyphenylazo)-uracil (HPUra), showed different effects on genetic recombination and DNA repair in Bacillus subtilis. Previous work (Pedrini et al., 1972) showed that NAL does not interfere with the transformation process of B. subtilis. The results reported in this work demonstrated that the drug was also without effect on the transfection by SPP1 or SPO-1 phage DNA (a process that requires a recombination event). The drug was also ineffective on the host cell reactivation of ultraviolet-irradiated SPP1 phage, as well as on transfection with ultraviolet-irradiated DNA of the same phage. HPUra instead markedly reduced the transformation process, as well as transfection, by SPO-1 DNA, but it did not affect the host cell reactivation of SPO-1 phage. In conclusion, whereas the NAL target seems to be specific for replicative DNA synthesis, the HPUra target (i.e., the DNA polymerase III of B. subtilis) seems to be involved also in recombination, but not in the excision repair process. The mutations conferring NAL and HPUra resistance used in this work were mapped by PBS-1 transduction.
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PMID:Effect of deoxyribonucleic acid replication inhibitors on bacterial recombination. 81 70

The amino acid metabolism of 23 different Brucella strains was investigated for differentiation purposes. The results were evaluated by thin layer chromatography, after enzymatic incubation. The organisms (Tab. 1) were grown on Tryptose blood agar at 37 degree C for 24 or 48h. Two mg wet weight of bacteria in 0.2 ml PBS, 0.01 M, were incubated with 12.5 microng (0.025 ml) amino acid in small tubes for 16h at 37 degree C, and centrifuged for 15 min at 7500 X g. For controls, bacterial suspensions were heated for 15 min at 100 degree C to destroy enzymatic activity, and also contrifuged for 15 min at 7500 X g. Usually 4 micronl of the supernatant fluids (6 micronl for L-asparagine, and 10 micronl for L-proline) were pipetted on the thin layer plate. The tests were run in n-butanol acetic acid water, 20:5:5, with a distance of 8 cm. Amino acids were stained with ninhydrine. The tests were repeated 3-5 times with identical results. Amino acid metabolism was indicated by different staining intesities (+ to ++) in comparison to control preparations. All species could be exactly differentiated from each other, with the exception of B. suis, biotype 2, and B canis, which could not be differentiated by their amino acid metabolism. Biotyps of the same species were mostly identical. The results of these investigations could be reproduced qualitatively as well as quantitatively. The method described is recommended for routine investigations.
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PMID:[Investigations of amino acid metabolism by thin layer chromatography for differentiation of Brucella (author's transl)]. 86 72


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