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)

Alcohol abuse increases the incidence and severity of opportunistic lung infections and pneumonias. Inducible nitric oxide (NO) synthase (iNOS II) and NO may be a pivotal system in the intracellular bactericidal activity of macrophages. We tested the hypothesis that acute administration of ethanol (ETOH) suppressed Escherichia coli endotoxin lipopolysaccharide (LPS) mediated upregulation of the iNOS II system in the lung of the rat, in vivo. We also tested the effect of ETOH on alveolar macrophage (AM) production of free NO using microelectrodes. Male Sprague-Dawley rats were given ETOH (5.5 g/kg, IP) 30 min. before giving intratracheal sterile phosphate buffered saline solution (PBS, 0.5 ml) or LPS (1 mg/kg in a total volume of 0.5 ml PBS). The isolated lungs were subjected to bronchoalveolar lavage (BAL) 3.5 hr. later. Aliquots of the BAL fluid were assayed for tumor necrosis factor alpha TNF alpha and reactive nitrogen intermediates (nitrate and nitrite) (RNI) with chemiluminescence. Aliquots of AM were incubated 1 hr ex vivo for spontaneous production of RNI or frozen and assayed for iNOS II mRNA with competitor exchange reverse transcriptase polymerase chain reaction (cERT-PCR). The lung was homogenized and assayed for RNI. LPS increased BAL fluid TNF alpha and RNI, lung RNI, and the spontaneous production of RNI by AM, ex vivo. These effects were inhibited by in vivo administration of inhibitors of iNOS II. LPS increased iNOS mRNA in AM. This was unaffected by iNOS inhibitors. ETOH suppressed LPS-induced BAL fluid TNF, iNOS mRNA and RNI production by AM and the lung.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Ethanol suppresses LPS-induced mRNA for nitric oxide synthase II in alveolar macrophages in vivo and in vitro. 753 15

Tumor necrosis factor-alpha (TNF alpha) and nitric oxide (NO) mediate in part the microbicidal response of murine and rodent alveolar macrophages (AM) and recruited neutrophils (PMN) to airborne infections. Ethanol (ETOH) suppresses intrapulmonary TNF alpha and NO release and impairs pulmonary host defense mechanisms. We tested the concept that ETOH down-regulates NO by inhibiting production of TNF alpha. Male rats were given intratracheal (i.t.) saline (PBS), a polyclonal anti-TNF alpha antibody (TNFab) or nonimmune IgG (22 mg/kg, i.m.) 2 h before giving i.t. Escherichia coli endotoxin (LPS) to normal rats or rats pretreated with ETOM (5.5 g/kg, i.p.) 30 min before experimentation. AM and PMN were obtained from the bronchoalveolar lavage fluid (BAL) fluid of rats killed 2 and 4 h after administration of LPS. mRNA for inducible NO synthase (iNOS) and TNF alpha were measured in AM and PMN with competitor equalized RT-PCR techniques. The BAL fluid, AM, and PMN were assayed for TNF alpha and NO2-, and NO3- (RNI) with the L929 bioassay and chemiluminescence, respectively. TNFab abolished LPS-induced increases in TNF alpha but did not suppress the NO content of the BAL fluid or gene expression for iNOS by AM or PMN. ETOH suppressed LPS-induced increases in mRNA for iNOS, production of RNI, and BAL fluid TNF alpha but did not affect LPS-induced increases in mRNA for TNF alpha. ETOH-induced attenuation of LPS-induced up-regulation of the iNOS system did not differ in rats pretreated with TNFab or IgG. Thus, ETOH down-regulates iNOS gene expression and RNI production independent of its effects on TNF alpha. Acute ETOH administration suppresses iNOS at the level of transcription and TNF alpha at the level of translation or release of the peptide.
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PMID:Independent suppression of nitric oxide and TNF alpha in the lung of conscious rats by ethanol. 754 Jan 57

The cobalt atom of hydroxocobalamin (OHC) binds cyanide and nitric oxide (NO) and OHC attenuates vascular responses to NO in vitro. NO mediates the hypotension of endotoxemia. Thus, we tested the postulate that OHC may attenuate the acute phase hypotension and toxicity associated with administration of Escherichia coli endotoxin (LPS). Rats were given OHC (20 mg/kg i.v.) or phosphate-buffered saline (PBS, 1 ml/kg i.v.) 30 min before or 15 min after giving LPS (0.8 mg/kg i.v.). Administration of OHC to PBS-treated control rats did not affect mean arterial pressure (MAP), heart rate or the plasma or urine content of the reactive nitrogen intermediates nitrate and nitrite (RNI). LPS decreased MAP by 50 mm Hg in PBS-treated rats and increased the plasma and urinary content of RNI. Administration of OHC to PBS-treated rats did not affect MAP or RNI. However, treatment with OHC before or after giving LPS attenuated LPS-induced hypotension and increases in plasma RNI and enhanced LPS-induced urinary excretion of RNI. OHC (20 mg/kg i.p.) or cyanocobalamin (10 mg/kg i.p.) given to Swiss-Webster mice 30 min before giving LPS (16 mg/kg i.p.) decreased the 24-hr mortality of LPS from 80 to 50% and the 36- and 96-hr mortality from 100 to 60% (OHC) or 70% (cyanocobalamin). Urine obtained from conscious rats given LPS (5 mg/kg i.p.) and OHC (20 mg/kg i.p.) exhibited a UV-visible absorbance spectrum with absorbance peaks characteristic of that formed after coincubation of NO and OHC.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Hydroxocobalamin (vitamin B12a) prevents and reverses endotoxin-induced hypotension and mortality in rodents: role of nitric oxide. 771 73

Recent food listeriosis outbreaks confirm that more faithful isolation and identification methods for Listeria monocytogenes or other potentially pathogen microorganisms are required. Furthermore, the human and animal reservoir role in the ecology of this disease must be established. Listeria spp. in the vizcacha intestinal content was determined by two isolation procedures, starting from 10 g of homogenized samples in 40 ml of PBS. I)0.1 ml was stripped on phenylethanol agar, selective agar for Listeria and acryflavin ceftazidin agar, then incubated at 37 degrees C for 48 h, suspected colonies were identified by preliminary tests (Gram, hemolysis, catalase, esculin hydrolisis and motility at 22 degrees C) and confirmatory tests (indol, methyl red, Voges Proskauer, nitrate and carbohydrate fermentation) (Table 1). Antibiotic susceptibility, protein profile by PAGE and pathogenic power in mice were determined. II) The remaining homogenate was incubated at 4 degrees C in 100 ml of Donnelly and Baigent enrichment broth, weekly or monthly with subcultures until 30 days or 6-8 months, respectively. The subcultures were followed up as in I). A L. seeligeri strain, susceptible to antibiotics suggested for L. monocytogenes and exhibiting resistance to some second and third generation cephalosporins, was isolated (Table 2). The protein profile of both species was coincident, but L. seeligeri was not virulent for mice. The finding of L. seeligeri in an animal (4.0%) used as human feeding source is of interest due to its potential pathogen power.
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PMID:[Isolation of Listeria seeligery from cecum of vizcacha (Lagostomus maximus maximus)]. 776 3

There is controversy regarding the evidence for the production of nitric oxide (NO) by neutrophils (PMNs). The present study investigates NO production, as assessed by the biosynthesis of the end products, nitrite and nitrate, in the pellets and supernatants of rat and mouse peripheral blood neutrophils obtained during endotoxemia and of peritoneal carrageenin-elicited PMNs stimulated in vitro with E. coli lipopolysaccharide (LPS). We also investigated the induction of NO synthase by rat and mouse peritoneal cells. The intraperitoneal (ip) administration of LPS to mice (10 mg/kg) and rats (5 mg/kg) significantly increased plasma nitrate concentration by six and 23-fold, respectively. In vivo pretreatment with L-NGmonomethyl arginine (L-NMMA) significantly inhibited this production. Compared to animals injected with PBS, the cell pellets of blood PMNs obtained from mice, but not rats, 2 or 6 h after LPS administration produced significant amounts of nitrite (14 +/- 3 and 18 +/- 2 nmol/mg protein, respectively). Little or no nitrite was found in the incubating medium. In contrast, 6 h after a carrageenin challenge (700 micrograms) peritoneal neutrophils obtained from rats, but not mice, released high concentrations of nitrite into the supernatant during a 24-h period of incubation (34 +/- 0.8 microM). The nitrite concentration of the pellet of these cells was negligible. In contrast to the lack of increase in the amount of nitrite released into the supernatants, the in vitro stimulation of rat PMNs with LPS (10 micrograms/ml) for 24 h did increase intracellular nitrite concentration (from 0.8 +/- 0.07 to 8 +/- 0.3 nmol/mg protein). In mouse PMNs, LPS treatment caused only a small release of nitrite into the incubation medium (14 +/- 1 microM). There was no significant change in nitrite concentration in the cell pellets. These data suggest that rat and mouse neutrophils differ in their ability to produce nitric oxide following stimulation with endotoxin.
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PMID:Differential production of nitric oxide by endotoxin-stimulated rat and mouse neutrophils. 873 34

Carvedilol (Kredex, Coreg) is a multiple action antihypertensive drug that has been shown to protect cell membranes from lipid peroxidative damages. In this study the physical and structural effects of carvedilol on lipid bilayers are investigated by fluorescence techniques, differential scanning calorimetry and other physical methods. Carvedilol binds to liposomal membranes (9:1 DMPC:DMPG) strongly with an apparent binding constant on the order of 10(4) M-1 in PBS (pH 7.4). The characteristic changes in its intrinsic fluorescence properties when bound to liposomes suggest that this compound is situated in a non-polar environment. The Stern-Volmer and bimolecular quenching constants, determined using nitrate as the fluorescence quencher, for the free and bound carvedilol indicate that the carbazole moiety is at a depth of > 11 A in the lipid bilayer. Fluorescence anisotropy measurements show that, unlike the membrane probes DPH and TMA-DPH, carvedilol is relatively mobile, and does not have a rigidly-defined molecular orientation in the bilayers. Differential scanning calorimetry results indicate that carvedilol is an effective membrane "fluidizer' as it dose-dependently lowers the gel to liquid crystalline transition temperature and broadens the endothermic transition. Comparative studies of interactions of carbazole, 4-OH carbazole and carvedilol with the model liposomal membranes reveal a possible role of membrane-partitioning in their antioxidant efficacy. These findings are discussed in perspective with the membrane biophysical properties of different classes of therapeutic significant lipid antioxidants in mind.
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PMID:Carvedilol-liposome interaction: evidence for strong association with the hydrophobic region of the lipid bilayers. 886 10

Ethanol (ETOH) inhibits the immune response to endotoxemia. The early stage of endotoxin (LPS)-induced shock is associated with an acute phase cardiovascular depression (APCD). Release of platelet activating factor (PAF) and tumor necrosis factor alpha (TNF alpha) with upregulation of nitric oxide (NO) production may initiate the APCD. Since ETOH inhibits induction of NO synthase (iNOS) mNRA by LPS, we postulate that ETOH may mask the APCD associated with endotoxemia. To test this, Sprague-Dawley rats (280-320 g, n = 5-6/group) were given LPS [0.75 mg/kg, intravenously (i.v.)] or PAF (10 to 150 micrograms/kg, i.v.) 30 min after administration of sterile saline (PBS), BN-5073 a mixed PAF antagonist (0.50 microgram/kg, i.v.), or ETOH [2.2-5.5 g/kg, intraperitoneally (i.p.)]. Cardiovascular parameters and plasma concentrations of nitrate and nitrite (RNI), ETOH, TNF alpha, and neutrophil (PMN) generation of RNI were measured. LPS and PAF both produced APCD. LPS-induced APCD was associated with tachycardia, elevated plasma TNF alpha and RNI, and ex vivo generation of RNI by PMNs. ETOH and BN-50730 prevented LPS-induced APCD and increases in RNI and TNF alpha. ETOH, however, increased the mortality associated with APCD. PAF produced only hypotension, bradycardia and elevated plasma levels of TNF alpha. ETOH and LNMMA did not affect PAF-induced APCD. BN-50730 inhibited PAF-induced APCD and plasma TNF alpha. We conclude that 1) ETOH inhibits the APCD and induction of NO characteristic of endotoxemia and 2) ETOH-induced suppression of LPS-mediated APCD may be mediated in part by suppression of release of intracellular PAF. Ethanol may increase the morbidity and mortality of endotoxemia by masking the hypotension and humoral changes characteristic of early endotoxemia thereby delaying appropriate therapy and by diminution of the protective effects of endogenous NO.
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PMID:Ethanol suppresses endotoxin but not platelet activating factor-induced hypotension and nitric oxide. 890 80

Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen that can be found in individuals in which the immune system has been suppressed by HIV/AIDS or chronic alcoholism. We evaluated the role of inducible nitric oxide synthase (NOS II) as a modulator of lung concentrations of P. aeruginosa in normal rats and rats given a single dose of ethanol (ETOH). Rats were pretreated with either sterile saline (PBS, 0.1 ml/kg, i.v.) or the NOS II inhibitor L-N6-iminoethyl lysine (LNIL, 10 mg/kg, i.v.) 15 min before intraperitoneal administration of either PBS (4.5 ml/kg) or ETOH (4.5 g/kg). Thirty min after administration of PBS or ETOH the rats were placed in inhalation chambers and exposed to 45 min of an aerosol containing P. aeruginosa (5 x 10(4) colony forming units, CFU). A group of rats (n = 5-6/treatment/time period) were killed immediately (0 hr) or 4 hr after inhalation of P. aeruginosa. The lungs were homogenized and the P. aeruginosa were grown in nutrient broth to determine the number of viable CFU remaining in the lung. The NOS II and TNFalpha mRNA and protein content lung alveolar macrophages (AM) and neutrophils (PMN) were measured with RT-PCR and Western blot. The concentration of nitrate and nitrite anion in the bronchoalveolar lavage fluid (BALf) and ex vivo incubates of PMN were also measured. The CFU of P. aeruginosa present in the lungs of the four groups of rats at 0 hr did not differ. The CFU of P. aeruginosa in the lung increased (p < 0.05) in rats pretreated with ETOH when compared with that obtained from rats pretreated with PBS. However, pretreatment of rats with LNIL decreased (p < 0.05) the 4 hr lung content of P. aeruginosa. Coadministration of LNIL and ETOH to rats augmented the CFU of P. aeruginosa in lungs to amounts which did not differ from that of rats pretreated with ETOH. Inhalation of P. aeruginosa increased NOS II mRNA and protein in rat AM and PMN. Pretreatment of rats with ETOH alone, or in combination with LNIL, inhibited P. aeruginosa-induced NOS II transcription and translation and AM and PMN nitrate and nitrite generation whereas pretreatment with LNIL alone only inhibited nitrate and nitrite generation. Pretreatment of rats with ETOH suppressed P. aeruginosa stimulated PMN recruitment into the lung whereas LNIL enhanced (p < 0.05) P. aeruginosa-stimulated PMN recruitment into the lung. ETOH-induced increases of the lung content of P. aeruginosa were associated with increased PKC delta isozyme in the membrane of the PMN but could not be explained by altered plasma concentrations of hydrocortisone or ETOH. The data demonstrate that selective inhibition of NOS II-derived NO by LNIL decreases the lung content of P. aeruginosa whereas ETOH inhibits the lung clearance of P. aeruginosa. Speculatively, the difference between these effects of LNIL and ETOH may result from differences in drug-induced changes in lung recruitment of PMN.
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PMID:Ethanol inhibits lung clearance of Pseudomonas aeruginosa by a neutrophil and nitric oxide-dependent mechanism, in vivo. 1023 11

The characterisation and selection of membranes by means of an immunofiltration assay is described. The chemical composition of the membranes was: nitro-cellulose, polyamide, polyvinylidene difluoride, polyethersulfone, cellulose acetate, regenerated cellulose, cellulose nitrate, and glass fibre. In order to characterise the membranes according to their binding capacity, immobilisation stability, sensitivity and hydrodynamic properties, two basic immunofiltration formats were performed. In both formats, enzyme label (horseradish peroxidase, HRP) and colorimetric detection were used. In the immobilised antibody format, three monoclonal antibodies (mAb) against the insecticide carbaryl were immobilised on the membranes by passive adsorption. In the immobilised hapten format, two haptens conjugated to bovine serum albumin (BSA) were immobilised. Immobilon-P was the best membrane with regard to the characterisation criteria and permitted the filtration of large volume (5.0 ml) through the membrane without release of the receptor. The immobilisation of the receptor (antibody or haptenic conjugate) was pH dependent. Good results with regard to mAb-antigen recognition, were obtained using 50 mM carbonate/bicarbonate buffer, pH 9.6. However, the most sensitive assays were achieved using, 10 mM phosphate buffer, 137 mM NaCl, 2.7 mM KCI (PBS), pH 7.4 as immobilisation buffer. Furthermore, all these results permit the choice of the best membrane for the rapid and sensitive determination of carbaryl. This study will assist the development of dipsticks, immunoelectrodes, membrane-based immunoreactors or immunoconcentration devices that are based on the use of membranes as immunosupports.
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PMID:Selection and characterisation of membranes by means of an immunofiltration assay. Application to the rapid and sensitive determination of the insecticide carbaryl. 1035 11

The direct electrochemistry of hemoglobin (Hb) immobilized on a hexagonal mesoporous silica (HMS)-modified glassy carbon electrode was described. The interaction between Hb and the HMS was investigated using UV-Vis spectroscopy, FT-IR, and electrochemical methods. The direct electron transfer of the immobilized Hb exhibited two couples of redox peaks with the formal potentials of -0.037 and -0.232 V in 0.1 M (pH 7.0) PBS, respectively, which corresponded to its two immobilized states. The electrode reactions showed a surface-controlled process with a single proton transfer at the scan rate range from 20 to 200 mV/s. The immobilized Hb retained its biological activity well and displayed an excellent response to the reduction of both hydrogen peroxide (H2O2) and nitrate (NO2-). Its apparent Michaelis-Menten constants for H2O2 and NO2- were 12.3 and 49.3 microM, respectively, showing a good affinity. Based on the immobilization of Hb on the HMS and its direct electrochemistry, two novel biosensors for H2O2 and NO2- were presented. Under optimal conditions, the sensors could be used for the determination of H2O2 ranging from 0.4 to 6.0 microM and NO2- ranging from 0.2 to 3.8 microM. The detection limits were 1.86 x 10(-9) M and 6.11 x 10(-7) M at 3sigma, respectively. HMS provided a good matrix for protein immobilization and biosensor preparation.
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PMID:Direct electron transfer and enzymatic activity of hemoglobin in a hexagonal mesoporous silica matrix. 1512 5


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