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)

The regeneration of antibodies covalently immobilized to an optical fibre surface was investigated by dissociation of the antibody-antigen complex with three different solvents: (a) an acidic solution (0.1 M glycine hydrochloride in 50% (v/v) ethylene glycol, pH 1.75), (b) a basic solution (0.05 M tetraethylamine in 50% (v/v) ethylene glycol, pH 11.0) and (c) 50% (v/v) ethanol in PBS. The fibres coated with polyclonal rabbit anti-goat antibody against a large protein retained 70% and 65% of the original signal after five consecutive regenerations with acidic and basic solvent systems, respectively. The fibres coated with monoclonal mouse anti-trinitrobenzene antibody specific for a small organic molecule, retained over 90% of the original signal when regenerated with basic and ethanol solutions. This study evaluated regeneration and reuse of antibody-coated fibre optic biosensors as a means of reducing routine laboratory analysis costs and time.
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PMID:Regeneration of immobilized antibodies on fiber optic probes. 782 82

The purpose of these experiments was to determine the effect of electroporation of IP3 into the cytosol of murine secondary oocytes and evaluate any alterations in [Ca2+]i resulting from Ca2+ release from intracellular stores. In addition, we evaluated the effect of ethanol (ETOH) on the release of Ca2+ from intracellular stores. Oocytes were loaded with the Ca2+ indicator fluo-3 by incubation in 100 microliters drops of medium containing 2 microM fluo-3/AM for 60 min at 37 degrees C. Changes in fluorescence were monitored by use of an inverted microscope which had been connected to a spectrofluorometer. Fluorescent intensity measurements were acquired for a minimum of 416 sec time span or up to 1,248 sec, with integration readings of 1 sec duration obtained every 2 sec throughout the measurement period. The experimental design consisted of comparing the rise in [Ca2+]i of fluo-3 loaded secondary oocytes subjected to electroporation in PBS and Ca(2+)-free PBS, each containing 25 microM IP3, to that elicited by PBS and Ca(2+)-free PBS containing a final concentration of 7% ETOH. Non-pulsed control secondary oocytes were placed in PBS + 25 microM IP3 during monitoring of [Ca2+]i fluorescence. Pulsed control secondary oocytes were placed in Ca(2+)-free PBS, subjected to electroporation pulse, and monitored for [Ca2+]i fluorescence. Electroporation of IP3 was accomplished by placing fluo-3 loaded secondary oocytes between the electrodes of a glass slide fusion chamber which had been overlaid with 300 microliters of PBS + 25 microM IP3 or Ca(2+)-free PBS + 25 microM IP3.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Electroporation of inositol 1,4,5-triphosphate induces repetitive calcium oscillations in murine oocytes. 842 41

The specific objective of this investigation is to study the effect of tricalcium phosphate delivery system (TCPL) particle sizes on the final density as well as the delivery profiles of various organic compounds in three different buffer environments. Each TCPL matrices were fabricated using three different particle sizes ranges between 1-38, 45-63 and 63-75 microns. The sintered microcrystal material was impregnated with either progesterone (P, 100 mg each) or bovine serum albumin (BSA, 100 mg each). In phase I of the study, each device was suspended in a serum bottle containing 100 mls of ethanol solution (50% wt/vol.) for P release or 100 mls of PBS (pH 7.4) for BSA release. In phase II, similar capsules were suspended in human plasma instead of standard buffers. The vials were agitated at 100 cycle per minute in a water bath set at 37 degrees C. The amount of P or BSA released from the devices into the buffered medium was measured spectrophotometrically. The results of this investigation revealed that a significant difference in the densities of the devices made from the range of individual particle sizes. The rate of steroid hormone and protein released from the devices made from 1-38 micron particle sizes was slower (p < 0.05) than the rate of delivery of P and BSA released from devices fabricated from either 45-63 or 63-75 micron particles. Regardless of the particle sizes effect the results show that the delivery profiles of BSA was higher than the rate of P. This observation could be attributed to the molecular structure as well as the physiochemical characteristics of the drug. In conclusion the data obtained from this study suggest that: (1) Particle sizes variations influence the density of the TCPL delivery system, (2) the rate of release of organic compounds from the ceramic devices is considerably affected by the physiochemical characteristics of medium or buffer system, and (3) the delivery rate of drugs from TCPL devices is directly proportional to the size of the device initial particles and macropores, and inversely proportional to the number of micropores within each device.
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PMID:Interrelationship between buffer systems and particle sizes of TCPL drug delivery system. 867 89

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.
Alcohol Clin Exp Res 1996 Oct
PMID:Ethanol suppresses endotoxin but not platelet activating factor-induced hypotension and nitric oxide. 890 80

The cysteinyl leukotriene LTE4 has been shown to induce airway eosinophilia in asthmatics in vivo. This phenomenon has not yet been reported for LTD4. Hence, we examined the effect of inhaled LTD4 and a control bronchoconstrictor agent, methacholine, on cell differentials in hypertonic saline-induced whole sputum samples of 12 nonsmoking atopic asthmatic subjects (three women, nine men; 21 to 29 yr of age; FEV1, 74 to 120% pred; PC20FEV1 methacholine < 9.6 mg/ml). The study had a cross-over, placebo-controlled design consisting of 4 d separated by > or = 1 wk. On each randomized study day, the subjects inhaled five serial doses of either LTD4 (mean cumulative concentration: 95.7 microM) or methacholine (mean cumulative concentration: 542 mM) or five doses of their respective diluents (PBS/ethanol or PBS). The airway response was measured by FEV1, followed by sputum induction with 4.5% NaCl, 4 h postchallenge. Inflammatory cells (> or = 250) were counted twice on coded cytospins and expressed as percentages of nonsquamous cells. There was no significant difference in the maximal percent fall in FEV1 from baseline between LTD4 (mean +/- SEM, 49.5 +/- 4.4% fall) and methacholine (mean +/- SEM, 55.9 +/- 3.4% fall) (p = 0.11). LTD4 induced a significant increase in the percentage of sputum eosinophils as compared with its diluent (mean +/- SD, 26.6 +/- 21.3% and 10.2 +/- 8.8%, respectively; p = 0.025), whereas a similar trend for methacholine failed to reach significance (mean +/- SD, 19.1 +/- 22.9% and 7.8 +/- 5.8%, respectively; p = 0.11). There was no significant difference in the changes in the percentage of sputum eosinophils between LTD4 and methacholine (mean difference +/- SD, 7.5 +/- 12.5% eosinophils; p = 0.09). We conclude that LTD4 induces eosinophilia in sputum of asthmatic subjects 4 h after inhalation. Our data suggest that LTD4 recruits eosinophils into the airways of asthmatics in vivo, possibly by virtue of direct or indirect chemotactic properties, whereas an additional effect of vigourous airway narrowing per se cannot be excluded.
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PMID:The effect of inhaled leukotriene D4 and methacholine on sputum cell differentials in asthma. 910 62

The bioactivity of feverfew (Tanacetum parthenium) leaf extracts has been analysed, by use of a human polymorphonuclear leukocyte (PMNL) bioassay, to assess the relative contributions of solvent extraction and parthenolide content to the biological potency of the extract. Extracts prepared in acetone-ethanol (system 1) contained significantly more parthenolide (mean +/- s.d. 1.3 +/- 0.2% dry leaf weight) than extracts in chloroform-PBS (phosphate-buffered saline; system 2; 0.1 +/- 0.04% dry leaf weight) or PBS alone (system 3; 0.5 +/- 0.1% dry leaf weight). Extract bioactivity, measured as inhibition of phorbol 12-myristate 13-acetate-induced, 5-amino-2,3-dihydro-1,4-phthalazinedione (luminol)-enhanced PMNL, chemiluminescence, followed a similar trend. Extracts inhibited phorbol 12-myristate 13-acetate-induced oxidative burst by amounts which, if solely attributable to parthenolide, indicated parthenolide concentrations for the respective solvent systems of 2.2 +/- 0.6%, 0.2 +/- 0.1% and 0.9 +/- 0.1% dry leaf weight. The mean ratio of parthenolide concentration to the parthenolide equivalent/PMNL-bioactivity value, for acetone-ethanol and PBS extracts were both 1:1.7. Parthenolide, although a key determinant of biological activity for T. parthenium leaf extracts based on the PMNL-bioassay, seems not to be the sole pharmacologically-active constituent. The identical and elevated bioactivity-parthenolide ratios for both organic and aqueons-phase leaf extracts suggest that a proportion of the other bioactive compounds have solubilities similar to that of parthenolide.
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PMID:Pharmacological activity of feverfew (Tanacetum parthenium (L.) Schultz-Bip.): assessment by inhibition of human polymorphonuclear leukocyte chemiluminescence in-vitro. 917 94

The study was performed to investigate the effect of penetration enhancers on the stratum corneum barrier. Epidermal membranes were prepared from freeze-stored (-70 degrees C) Caucasian breast skin and mounted in a flow-through diffusion cell. The validity of the freeze storage procedure was verified by measurement of [3H]-water penetration. The effect of the cyclic terpene, carveol, on the transdermal penetration of water and ethanol was studied in vitro. Control ethanol and water penetration measured with a donor solution of 50% ethanol/PBS (w/w) was 1.9+/-0.2 and 3.6+/-0.5 x 10(-3) cm/h. The addition of 3% carveol to the donor solution increased the permeation of ethanol and water after 4 h to 8.3+/-1.1 and 12.5+/-1.9 x 10(-3) cm/h, respectively. In a separate experiment, terpinen-4-ol and alpha-terpineol were also tested, in addition to carveol, for effect on tritium flux. No significant difference in maximum tritium flux was obtained between the three terpenes studied. The maximum increase in permeability coefficients of carveol, terpinen-4-ol and alpha-terpineol was 10.6, 8.7 and 10.9, respectively.
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PMID:Terpene-enhanced transdermal permeation of water and ethanol in human epidermis. 922 15

Mammalian oocytes can be induced to resume meiosis without fertilization, and the resulting parthenogenetic embryos carry only maternal chromosomes. Human oocytes can be activated by many chemical and physical stimuli, but postimplantation studies of human parthenogenetic embryos are not ethically acceptable. The common marmoset monkey (Callithrix jacchus) is a good model for studying primate parthenogenetic development postimplantation, since follicular aspiration, embryo transfer, and early postimplantation development of biparental embryos have already been described. Marmoset oocytes were either subjected to two series of six electrical pulses (DC; 2 kV/cm and 70 microsec) or were incubated in 7% ethanol in PBS. Ninety-two percent (68 of 74) and 20% (8 of 40) of marmoset oocytes were activated by electrical stimulus or ethanol, respectively. Parthenogenetic (n = 3) or in vitro-fertilized (n = 2) embryos were transferred at the 4-cell stage to synchronized recipient female marmosets (n = 5). Progesterone, chorionic gonadotropin, and inhibin in the peripheral plasma of recipient animals were measured. After 33 days of gestation, recipient animals were perfused and the uteri were collected. The 2 females that had received biparental embryos and 2 of the 3 females that had received parthenogenetic embryos displayed biochemical and histological evidence of implantation. This is the first report that a primate embryo comprising only parthenogenetic cells is capable of implantation. This highlights the need to scrutinize levels of parthenogenesis associated with human assisted reproductive technologies. Marmoset parthenogenones also provide a unique model for elucidating the roles of parental genomes in primate development.
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PMID:Parthenogenetic activation of marmoset (Callithrix jacchus) oocytes and the development of marmoset parthenogenones in vitro and in vivo. 982 97

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.
Alcohol Clin Exp Res 1999 Apr
PMID:Ethanol inhibits lung clearance of Pseudomonas aeruginosa by a neutrophil and nitric oxide-dependent mechanism, in vivo. 1023 11

An ascorbic acid decalcifying solution was applied to immuno- and affinohistochemical studies on the inner ear. Rat inner ears fixed in 4% paraformaldehyde in PBS or in 2% acetic acid in ethanol solutions were adequately decalcified in an ascorbic acid solution, at a temperature of 4 degrees C. The decalcifying solution was prepared with 1% ascorbic acid and 0.84% sodium chloride in distilled water (pH 2.5-2.6). The decalcification time was in a direct relationship to the specimen calcification. In this study, two neuroactive substances (gamma-aminobutyric acid and calcitonin gene-related peptide), neurofilaments, and the galectine endogenous lectin were successfully detected immunohistochemically.
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PMID:Decalcification by ascorbic acid for immuno- and affinohistochemical techniques on the inner ear. 1046 Apr 65


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