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)

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

Alloxan participation in extracellular redox processes results in the formation of the reactive oxygen species (ROS) superoxide anions (O2-), hydroxyl radical (OH.) and hydrogen peroxide (H2O2), causing cell damage through a number of complex interactions probably involving several different cellular structures. These involve the plasma membrane, and we have recently presented evidence for lysosomal interference. The present study elucidates the early (within 15 min) events in a model system of macrophage-like cells (J-774) in culture. Addition of 2 mM alloxan and 1 mM cysteine to the medium surrounding the cells (phosphate-buffered saline, PBS, 37 degrees C, pH 7.4) resulted in rapid lysosomal membrane damage with disappearance of the proton gradient as visualized by acridine orange relocalization, as well as plasma membrane alterations leading to increased leakage of fluorescein after fluorescein diacetate staining. These events were later (greater than 30 min) followed by cellular degeneration in the form of blebbing. Mitochondrial damage (rhodamine 123 relocalization) was a late event. Cells pretreated with desferrioxamine (Des) and superoxide dismutase (SOD) or Des, SOD and catalase (CAT) to induce partial (H2O2 formation only) or almost full protection (no ROS formation) showed about the same reactions as when cells were exposed to alloxan and cysteine without scavengers (O2-, H2O2 and OH. formation) or with PBS only, respectively. The results are interpreted as indicating that the cytotoxicity is a consequence mainly of H2O2 involvement and probably of lysosomal influx of H2O2 with ensuing OH.formation within secondary lysosomes containing trace amounts of reactive iron. It is suggested that the resultant lysosomal membrane damage is followed by leakage of lysosomal hydrolases and ensuing cellular degeneration.
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PMID:Extracellular reduction of alloxan results in oxygen radical-mediated attack on plasma and lysosomal membranes. 158 Oct 40

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

Estradiol (E2) facilitates the lordosis reflex that occurs in response to flank stimulation in female rats. Lordosis appears to be regulated in part by the synthesis of proteins in the ventral medial hypothalamus (VMH) that are transported to the midbrain central gray (MCG). We developed a strategy involving microinfusion of radioactive amino acids, followed by 2-dimensional gel electrophoresis, to identify proteins that may be regulated by E2 in the VMH and transported to the MCG. A mixture of 35S-methionine and 35S-cysteine (2:1, total 500-1000 microCi), suspended in 1 microliter PBS, was infused bilaterally into the VMH over a period of 2 hr into matched pairs of ovariectomized female rats, one of which was given a Silastic implant containing E2 at the beginning of infusion or 1 week earlier. The rats were sacrificed 12 hr after the end of infusion, and several brain regions were obtained by microdissection. Samples were analyzed by 2-dimensional gel electrophoresis, entailing isoelectric focusing in the first dimension and SDS-PAGE (molecular-weight separation) in the second dimension, followed by fluorography. We could routinely separate at least 250 spots. We consistently found a protein spot with an apparent molecular weight of 70 kDa, pI of about 5.9, which almost always appeared in the VMH and MCG of rats given E2 replacement but very rarely in samples from ovariectomized rats given no E2 replacement. A spot immediately acidic to this protein (70 kDa, pl about 5.8) appeared to vary inversely with this E2-induced protein.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:An estradiol-induced protein synthesized in the ventral medial hypothalamus and transported to the midbrain central gray. 282 60

A dose-dependent and transiently elevated expression of a cytoplasmic 32 kDa protein was observed in Swiss albino 3T3 fibroblasts exposed to mainstream cigarette smoke (CS) trapped in phosphate-buffered saline solutions (smoke-bubbled PBS). The protein was identified as heme oxygenase (HO) (heme, hydrogen donor:oxygen oxidoreductase, EC 1.14.99.3) by Western blotting using an anti-rodent HO-specific antibody. Kinetic investigations revealed that HO protein and its mRNA were detectable in smoke-bubbled PBS-treated cells between 1 and 24 h after exposure to 0.03 puffs (approximately 1 cm3) CS per ml medium. As a result of transcriptional activation, a nearly 50-fold increase in the amount of HO mRNA was determined after 8 h exposure compared to control levels. Since literature data indicate that there is a link between glutathione depletion and HO expression, the same was assumed for cells exposed to smoke-bubbled PBS, as a decrease of more than 60% in glutathione levels was observed after the exposure. This was further supported by the observation that no elevated amounts of HO mRNA appeared in smoke-bubbled PBS-treated cells when cysteine was exogenously added. However, although these effects may be attributable to the formation of hydroxyl radicals (which have been shown to induce HO and to deplete glutathione levels and which appear in aqueous smoke-containing solutions via the iron-catalysed Fenton reaction) neither catalase nor the iron cation chelating agent o-phenanthroline were able to suppress or even to reduce HO expression in smoke-bubbled PBS-treated cells. On the contrary, at comparable concentrations both compounds were found to be potent inhibitors of smoke-dependent DNA strand breaks. Hence, reactive species other than Fenton reaction-derived hydroxyl radicals are responsible for the effects observed in the present study.
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PMID:Heme oxygenase expression in Swiss 3T3 cells following exposure to aqueous cigarette smoke fractions. 829 50

Helicobacter pylori cells cultured on solid medium were quantitatively tested for haemolytic activity against erythrocytes of man, sheep, the guinea pig and rabbit. Using 4-day and 8-day cultures of two standard strains (ATCC 43504, IMMi 676), human erythrocytes were not lysed by 10% bacterial suspensions. Rabbit erythrocytes were the most sensitive to 8-day cultures. Hot-cold incubation yielded the highest haemolysis titres. The extent of haemolysis strongly correlated with the number of bacterial cells. Supplementation of the test medium (PBS, pH 7.4) with L-cysteine, dithiothreitol, MgCl2, EDTA, cholesterol, lecithin or sphingomyelin did not influence the haemolysis titres. They were significantly reduced in the presence of pronase E, human serum, bovine serum albumin or CaCl2, and by heat treatment of the bacteria. Supplementation of the test medium with cardiolipin strongly increased the haemolysis titres. Comparing the cell-associated haemolytic activity of 18 strains, the titres ranged from < 2 to 64, with a median titre of 16. No correlation was found between the haemolytic activity and phospholipase C activity of the cell suspensions. It was concluded that the formation of lysophosphatides and non-enzymatic factors rather than a sulphydryl-activated cytolysin or phospholipase C are responsible for the cell-associated haemolytic activity. This property may be involved in the pathogenicity and virulence of Helicobacter pylori.
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PMID:Cell-associated haemolytic activity of Helicobacter pylori. 850 Apr 89

To determine optimal conditions for allergen preservation, we investigated the influence of different stabilizing additives and of storage temperature on the allergen activity of apple protein preparations, obtained by extraction in phosphate buffer or by precipitation in diacetone alcohol and resolubilization in phosphate buffer in the presence or absence of enzyme inhibitors. For this purpose, the extracts were stored for 6 months either in frozen state at -20 degrees C or in lyophilized state at -20 degrees C, 4 degrees C, or room temperature and were characterized by SDS-PAGE, immunoblot, ELISA inhibition, and prick test. The highest stability revealed the extracts that were prepared by precipitation in the organic solvent in the presence of enzyme inhibitors, lyophilized, and stored at -20 degrees C. For storage of extract solutions at 4 degrees C, PBS/glycerol and cysteine/sodium citrate/glycerol were found to be the most effective stabilizing additives.
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PMID:Investigation of the stability of apple allergen extracts. 858 90

There were two related objectives in this study. The first was to determine the influence of endogenous relaxin on ovulation in rats. The second was to investigate the effect of relaxin on the secretion of gelatinases involved in extracellular matrix remodeling from rat ovarian cells. Immature rats were primed s.c. with 10 IU eCG; 51 to 52 h later, a monoclonal antibody specific for rat relaxin (MCAR), a control antibody against fluorescein (MCAF), or PBS vehicle was administered via intraovarian bursal injection under anesthesia, and 15 IU hCG was injected i.p. immediately thereafter. Rats were killed 26 h later, and oviducts were isolated and examined under the microscope to determine the number of ovulated oocytes. MCAR (0.25 and 2.5 micrograms/ovary) partially suppressed gonadotropin-induced ovulation as compared to the value for PBS controls. There was no significant difference in the number of ovulated oocytes between animals treated with MCAF and PBS controls. Also, porcine relaxin, given s.c. immediately after MCAR treatment, could reverse the inhibitory effect of MCAR on ovulation. To examine a possible mechanism for the effect of relaxin on ovulation, granulosa cells and theca-interstitial cells were obtained from ovaries of eCG-primed immature rats. The gelatinases secreted from cultured cells were analyzed using gelatin zymography and scanning densitometry. In the granulosa cell culture, relaxin increased the secretion of two major gelatinases of about 92 and 63 kDa in a dose-and time-dependent manner within 24 h of treatment. In the theca-interstitial cell culture, relaxin induced dose- and time-dependent increases in the secretion of two other major gelatinases of about 76 and 71 kDa. These gelatinases were characterized as metalloproteinases but not serine/cysteine proteinases. Furthermore, an immunoblot study demonstrated that relaxin stimulated the secretion of a 72-kDa type IV collagenase-like substance from cultured theca-interstitial cells but not from granulosa cells. This study demonstrates several original findings. First, endogenous relaxin may facilitate the ovulatory process in rats. Second, exogenous relaxin exhibits a biological effect on cultured rat theca-interstitial cells in addition to granulosa cells. Third, exogenous relaxin regulates the secretion of different major forms of gelantinases from cultured rat granulosa cells and theca-interstitial cells. The study supports the idea that relaxin may play an autocrine/paracrine role that is involved in modulating ovarian function.
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PMID:Relaxin modulates the ovulatory process and increases secretion of different gelatinases from granulosa and theca-interstitial cells in rats. 894 84

The effects of methylthio-cysteine disulfide (MT-Cy) and cystamine (CAM) on the thiol production and glutathione content of a human T cell line (CEM-SS) have been investigated. MT-Cy per se and CAM in the presence of cystine greatly enhanced thiol production and glutathione content of cells while cystine alone exerted no or slight influence in the first hours. The MT-Cy- or CAM-induced extracellular SH-generation was observed both in a complete nutrient medium and even more in SH-free D-PBS. The acid-soluble thiol level and glutathione content of cells elevated markedly (up to 5-6 fold in two hours) when incubating cells in complete medium. Inhibition of glutathione synthesis by DL-buthionine (S,R)-sulfoximine did not alter the MT-Cy- or CAM-induced extracellular thiol production indicating that glutathione synthesis is not involved in this effect. The results suggest that MT-Cy easily enters the cells thus accelerating the thiol cycle in SH-poor medium while CAM promotes cystine uptake into the cells. Phenylalanine and leucine inhibited both MT-Cy- and CAM-dependent thiol production in D-PBS most effectively suggesting the involvement of the L membrane transport system in these effects.
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PMID:S-methylthio-cysteine and cystamine are potent stimulators of thiol production and glutathione synthesis. 948 18

Mainstream cigarette smoke (CS) trapped in phosphate-buffered saline solutions (smoke-bubbled PBS) has been shown to induce a strong stress response in cultured cells. This is reflected, for example, by the expression of stress genes such as c-fos and haem oxygenase, a transient decrease in the translation efficiency and the induction of cell cycle arrest. In these studies, peroxynitrite, the reaction product of nitric oxide (NO) and superoxide (O2-.), was identified as an active principle formed by CS in aqueous solutions. In the present study, we show that the CS-induced stress response is critically dependent on the intracellular glutathione (GSH) content which itself becomes diminished in cells exposed to smoke-bubbled PBS. Investigations using c-fos expression as a measure for cellular stress revealed a direct correlation between the smoke-bubbled PBS concentration necessary for stress-dependent c-fos expression and the intracellular GSH concentration observed in different cell lines. Correspondingly, 3T3 fibroblasts artificially depleted of GSH by pretreatment with buthionine-sulphoximine (BSO), an inhibitor of GSH synthesis, require significantly lower amounts of smoke-bubbled PBS to obtain a detectable c-fos expression, whereas, supplementation of the medium with N-acetyl-cysteine is an efficient treatment for the inhibition of a CS-induced c-fos response. We also show that the smoke-bubbled PBS-dependent loss of intracellular GSH is mainly attributable to the aldehyde fraction of CS, although these aldehydes by themselves cannot induce c-fos in these cells. The smoke-bubbled PBS-dependent c-fos response can, however, be mimicked when peroxynitrite and CS-related aldehydes, at the concentrations calculated to appear in smoke-bubbled PBS, are used in combination for cell exposure. Taken together, these results suggest that in cells exposed to aqueous extracts of CS, smoke-related aldehydes decrease the intracellular GSH content significantly, allowing peroxynitrite to interfere with specific target molecules resulting in the stress-specific expression of c-fos.
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PMID:The cellular stress response induced by aqueous extracts of cigarette smoke is critically dependent on the intracellular glutathione concentration. 963 65


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