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

Several mucolytic agents were evaluated on sputum for testing their viscolytic activity and the bacterial tollerance to each of them. Proteolytic enzymes (trypsin, pepsin, papain, pancreatin), KJ, and dithiothreitol (or its derivatives) were better tollerated by common respiratory pathogens (H. influenzae, D. pneumoniae, Klebsiella, etc.) than other mucolytic agents, as acetil-cysteine, cisteamine-HCl, tension active substances, mercaptoethanol, and others. The dithiothreitol showed also one of the strongest viscolytic effect and therefore it was selected for the routinary sputum digestion at the concentration 0.1% in PBS pH 7.2. Such a solution was added to sputum specimen in different proportions according to the macroscopic "apparent" viscosity of each specimen. However researches on the comparative viscolytic activity of all the agents hereinafter considered are still in progress.
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PMID:[Study on the viscolytic activity of the sputum (author's transl)]. 1 42

A method is described for producing banding patterns with methyl green-pyronin (MGP) stain in chromosomes of fibrosarcoma cells. 1) The stain was made by mixing equal volumes of 2% aqueous pyronin G, 2% aqueous methyl green, distilled water, and 0.1 M acetate buffer (pH 5.7). 2) Treatment with colcemide and hypotonic KCl (0.075 M) was performed as usual. 3) Metaphase chromosomes were prepared using the flame-drying technique and treated with 0.25% trypsin at 37 C for 45 to 90 seconds. Before staining, the slides were rinsed in PBS, in distilled water, and then were dipped in 0.05 M acetate buffer. 4) Chromosomes were stained for more than 20 minutes, rinsed in distilled water, and hot-air dried. Satisfactory results were obtained in uncontracted metaphase chromosomes. MGP stain has the advantage of permitting much longer trypsin treatment and staining time than the trypsin-Giemsa method while providing satisfactory banding patterns.
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PMID:Chromosomal banding patterns produced by methyl green-pyronin staining after trypsin treatment. 49 34

A simple method for the preparation of a potent group-specific antigen on HeLa-229 cells infected with MRC-1 (LB) (TRIC/GB/MRC-1 Gf) strain of Chlamydia trachomatis is outlined. HeLa-229 cells are infected (MRC-1 strain, 102 inclusion-forming units per cell) with centrifugation at 4,000 g for 1 h in flat-bottomed vials. The cells are removed by brief trypsinization with 0.25% trypsin and put into 75 cm2 culture flasks (12 x 106 cells by flask) in BHK-21 medium supplemented with foetal bovine serum. The flasks are incubated for 5 days at 37 degrees C. The destroyed cell monolayer and the supernatant are centrifuged at 100,000 g for 1 h. The pellet is collected and resuspended in PBS and subjected to ultrasonic vibration for 30 min. This antigen may be used for detection of complement-fixing antibodies in lymphogranuloma venereum and ornithosis.
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PMID:[A method for the preparation of a Chlamydiae group-specific antigen on hela-229 cells infected with a strain of "Chlamydia trachomatis" for use in the complement fixation test (author's transl)]. 53 72

A method of separating lymphoid cells from solid mouse mammary tumors was developed and evaluated. In this method the tumors are digested with 0.01% collagenase, 0.01% DNAase, and 0.025% trypsin in Dulbecco's PBS into suspensions of cells with a viability of 90%. The suspensions are fractionated on a continuous gradient of Ficoll in tissue culture medium. In model experiments this gradient was found to separate, cleanly, admixed cells of an established mammary tumor cell line and dissociated thymus glands. Recovery rates were 50% for the tumor cells and 80% for the thymocytes. The preparation of the cell suspensions and the gradient separation procedure are not harmful to the cells as indicated by trypan blue exclusion and the ability to grow in cell culture.
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PMID:In situ lymphoid cells of mouse mammary tumors. I. Development and evaluation of a method for the separation of lymphoid cells from mouse mammary tumors. 65 81

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

Sciatic nerves from mice were removed and soaked in either PBS (phosphate buffered saline) or PBS plus I% trypsin (Sigma Type III) for various periods of time. Specimens were soaked at either room temperature or 37-degrees C at pH's ranging from 7.5 to 8.0. The epineural and perineural sheaths were split to allow the trypsin to penetrate the nerve. Tissue was prepared for electron microscopy by fixation in cacodylate buffered formaldehyde-glutaraldehyde solutions, post-fixed in OSO4 and embedded in Epon 812 or in glutaraldehyde-urea resin without osmication. After four h incubation at 37-degrees C or eight h at room temperature, the basement membranes of the Schwann cells became fragmented and detached and the myelin intraperiod band lost some density. After 18 h, myelin with swollen intraperiod bands displaying a loss of electron density and split main period bands was noted adjacent to normal myelin. Other areas had been transformed into vesicles indicating that the membranes of these vesicles appeared to have been derived from the detachment of both the intraperiod and main period bands within the myelin. Evidence is presented for the presence of trypsin digestable proteins in both the main period and intraperiod bands of peripheral nervouse system myelin.
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PMID:Electron microscopy of trypsin-digested peripheral nerve myelin. 111 38

Immunohistochemical analyses have shown the presence of T lymphocytes (T-cells) in atherosclerotic places in addition to macrophages and smooth muscle cells. To elucidate the role of T-cells in the formation of atherosclerotic lesions, we studied whether T-cells can stimulate the scavenger pathway and promote esterified cholesterol (EC) synthesis by [14C]oleate incorporation in macrophages. Macrophages and T-cells were co-cultured in two ways. In one culture, macrophages were in direct contact with T-cells (direct contact form). In the other, macrophages and T-cells were separated by Transwell membrane, but shared the same culture medium via the membrane (indirect contact form). Based on the incorporation of [14C]oleate into EC, macrophages strikingly increased EC synthesis in both forms of co-culture. This increase was proportional to the number of T-cells present and was inhibited by cyclosporin A. When macrophages were co-cultured indirectly in contact with T-cells in the presence of AcLDL for 24 h, and the T-cells were subsequently removed, EC synthesis in macrophages increased. However, this increase was not observed in macrophages that were rinsed twice with PBS. When macrophages, previously incubated with AcLDL for 24 h, were co-cultured indirectly in contact with T-cells for 24 h, the medium were prepared as activated T-cell-conditioned medium (aTCM). EC synthesis in macrophages cultured with aTCM increased. The ability of aTCM to increase EC synthesis disappeared upon repeated freezing/thawing, boiling and trypsin treatment. T-cells (indirect contact form) and aTCM similarly increased AcLDL-binding and -degradation in macrophages. These results indicated that T-cells secreted an active substance(s), protein in nature, which could activate the scavenger pathway and increase EC synthesis in macrophages. These observations suggest that T-cells can promote the uptake of modified lipoproteins by macrophages to induce foam cell-formation.
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PMID:T lymphocytes increase the synthesis of esterified cholesterol in human monocyte-derived macrophages by activation of the scavenger pathway. 156 18

The investigations were carried out on 59 cows from Holstein half-breed, establishing that 8 cows suffered salpinx obstruction (5 cases with unilateral obstruction and 3 cases with bilateral obstruction). The authors are using an apparatus made by themselves, for insufflation of air in the obstructed uterus, and which is useful in desobstruction treatment, too. For diagnosis, CO2 was introduced inside of uterus. The authors used for treatment PBS (saline phosphate buffer) in addition with penicillin G, hydrocortisone and trypsin. Before air insufflation in uterus there will be infused 10-20 ml 2% Lidocaton. The cows must be examined in oestrus period, or 2 days after PGF2 alfa administration. The gas must be introduced under rectal palpation, and pressure must not be higher than 500 mm H2O column. If there is a permeable oviduct, after 15-20 sec. from gas introduction, ist is possible to palpate the filled oviduct. From ovary we can perceive a rustle produced by gas crossing in abdominal cavity. In case of salpinx obstruction, the treatment must be start as soon as possible. The utilized liquid for treatment will be introduce by gas pressure, inside of uterus and oviducts. Using this method, it managed the repermeability of oviducts at 3 from 8 treated cows. In each case, there were used 3 treatments at 48 h interval. After the second insemination (I.A.) 2 cows remained pregnant.
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PMID:[A method for the treatment of oviduct obstruction in cattle using CO2 insufflation into the uterus]. 175 12

P. gingivalis adheres to A. viscosus on mineral surfaces mimicking teeth. To study whether P. gingivalis proteases contribute to its binding, mutants of P. gingivalis deficient in proteases were compared with their parent strain and a P. gingivalis-type strain for their adherence to A. viscosus on saliva-coated hydroxyapatite by manipulating a radio-isotope binding assay. Adherence of P. gingivalis 2561 to A. viscosus was studied by tests of the effects of incubation temperature and known inhibitors or promoters of proteases. Controls were handled by the assay run in PBS buffer at 22 degrees C. Two mutants deficient in trypsin-like protease were found to be deficient in adherence (% attachment relative to control: 3.2 +/- 0.1% and 11.2 +/- 0.4%), while a collagenase-deficient mutant had an adherence score (51.6 +/- 8.4) closer to that of the parent strain (75.6 +/- 7.2%). Heating P. gingivalis at 70 degrees C decreased its subsequent adherence at 22 degrees C by 80%. Adherence decreased by 60% when the assay was run at 4 degrees C, but increased by 70% at 37 degrees C. Reducing agents (dithiothreitol, cysteine, and mercaptoethanol) enhanced P. gingivalis adherence by 50 to 60%. Protease inhibitors (BZMD, SBTI, TPCK, TLCK, CMPS, PMSF) decreased adherence by 10 to 50%. Also, Hg2+ and Zn2+ decreased adherence by 30 to 50%, and arginine decreased it by 50%. Most of these effects on P. gingivalis adherence were statistically significant (p less than 0.05). Analysis of these data suggests that P. gingivalis proteases may contribute to the cohesion of P. gingivalis and A. viscosus.
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PMID:Association of proteases of Porphyromonas (Bacteroides) gingivalis with its adhesion to Actinomyces viscosus. 184 87

Five murine monoclonal antibodies (Mab's) have been produced which are reactive with the surface of the third-stage larvae (L3) of Onchocerca volvulus. These were produced from a fusion performed after intrasplenic injection of 10 live O. volvulus L3. Hybridomas were first screened by Elisa using a PBS extract of O. volvulus female adult worms. Elisa negative wells were screened by IFA on whole formalin-fixed L3. Five Mab's were isolated which were reactive with the surface of L3, all were found to be of the IgM isotype. All five Mab's were cross-reactive with the surface of O. lienalis L3, but not with the L3 of Brugia malayi, B. pahangi, Dirofilaria immitis, or Acanthocheilonema viteae. One of the Mab's, OV3-9, reacted with an antigen common to many species of helminths. The other four were genus specific reacting only with O. volvulus and O. lienalis. All 5 Mab's reacted with L2 of O. volvulus, but not with the L4 by IFA on whole fixed larvae. Only Mab's OV3-8 and OV3-9 reacted with cryosections of adult O. volvulus by IFA indicating that the other 3 Mab's are specific for the surface of L2/L3 of the genus Onchocerca. None of the Mab's reacted with detergent dissociated L3 proteins in Western Blot analysis. The ability of L3 to bind the Mab's was abolished by incubating larvae in either proteinase K or trypsin but was unaffected by incubation of larvae in detergents, or by treatment with periodate, indicating the proteinaceous nature of the Mab specific epitopes. Anti-phosphorylcholine Mab (Gib-13) did not react with the surface of L3.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Onchocerca volvulus: characterization of monoclonal antibodies reactive with surface components of third-stage larvae. 189 79


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