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J A Mitchell

Publications and source records attributed to J A Mitchell.

At least 163 records · Page 9Linked to original sources
PubMed1992

Nitric oxide synthase in ferret brain: localization and characterization.

1. In the present study, we have investigated the distribution of nitric oxide synthase in the ferret brain. Nitric oxide synthase was determined biochemically and immunochemically. 2. In the rat brain, the highest nitric oxide synthase activity has been detected in the cerebellum. However, in the ferret brain, the highest activity was found in the striatum and the lowest in the cerebellum and cerebral cortex. The enzymatic activity was localized predominantly in the cytosolic fractions, it was dependent on NADPH and Ca2+, and inhibited by NG-nitro-L-arginine or NG-methyl-L-arginine. 3. Western blot analysis revealed that all regions of the ferret brain contained a 160 kD protein crossreacting with an antibody to nitric oxide synthase purified from the rat cerebellum, and the levels of relative intensity of staining by the antibody correlated with the distribution of nitric oxide synthase activity. 4. These results indicate that the ferret brain contains a nitric oxide synthase similar to the rat brain, but the distribution of enzymatic activity in the ferret brain differs markedly from the rat brain.

Amino Acid Oxidoreductases↗
PubMed1992

Effects of alcohol on intrauterine oxygen tension in the rat.

The effects of alcohol on the availability of oxygen within the uterine lumen of rats were determined on Day 4 of pseudopregnancy. Intraluminal oxygen tension (pO2) was measured in vivo in anesthetized rats using a 22-gauge polarographic oxygen sensor. Intrauterine pO2 was measured for 15 min before and after intravenous injection of alcohol (1.0 g/kg: 100% ethanol/saline, 1:2v/v) or vehicle (physiological saline). Alcohol administration increased mean intrauterine pO2 (mm Hg) from a pretreatment level of 28.3 +/- 2.8 to 38.7 +/- 3.8 mm Hg (p less than 0.05, n = 10) at 15 min postinjection. The rapid rise in oxygen tension was accompanied by increased frequency of fluctuation (peaks/hr) in intraluminal pO2 (prealcohol: 64.2 +/- 8.8 vs. postalcohol 96.0 +/- 7.7 peaks/hr; (p less than 0.05, n = 10). Injection of saline did not alter any aspect of intrauterine pO2. During the period of measurement of luminal pO2, blood alcohol levels increased from 0 during pretreatment to 106 mg% within 10 min of injection. These results indicate that alcohol increases the availability of oxygen within the uterus during the time of endometrial sensitivity to deciduogenic stimuli and blastocyst implantation.

Animals↗
PubMed1992

Comparison of the release of vasoactive factors from venous and arterial bovine cultured endothelial cells.

The release of endothelium-derived relaxing factor (EDRF), prostacyclin (PGI2), and endothelin-1 (ET-1) was measured from endothelial cells (EC) cultured from either bovine vena cava (BVCEC) or bovine aorta (BAEC). EDRF release was determined by using the superfusion bioassay technique, whereas ET-1 and PGI2 were measured by specific radioimmunoassays. Bradykinin (BK) (0.05-30 pmol) given through columns of venous or arterial EC induced a dose-dependent release of EDRF. BK (0.05 pmol) evoked a release of EDRF from venous EC that was similar to the effect of a dose of 1 pmol from arterial EC. As with BAEC, infusions of NG-monomethyl-L-arginine (30 microM) caused an inhibition of EDRF release from BVCEC that was partially reversed by coinfusions of L-arginine (L-Arg; 100 microM). BK also induced a dose-dependent release of PGI2 from BVCEC. BVCEC and BAEC produced PGI2 in equivalent amounts when arachidonic acid (9.2 and 32 pmol) was added to the Krebs' solution perfusing the cells. BVCEC and BAEC released detectable amounts of ET-1 (0.4 +/- 0.1 and 0.9 +/- 0.3 ng/mL, respectively), over a 4-h period, and the release of ET-1 was increased approximately twofold by coincubations with thrombin (0.05-1 U/mL). These findings demonstrate that venous EC have a similar capacity to arterial EC to release vasoactive factors, thus supporting the hypothesis that veins have a functional endothelium that may modulate venous tone and platelet function.

Amino Acids↗
PubMed1992

Differential ability of occupational chemical contact and respiratory allergens to cause immediate and delayed dermal hypersensitivity reactions in mice.

Trimellitic anhydride (TMA) is known to cause occupational respiratory allergy associated with the presence of specific IgE antibody. Other chemicals, such as 2,4-dinitrochlorobenzene (DNCB), while exhibiting a clear potential for contact sensitization, apparently lack the ability to induce respiratory allergy in man. It has been shown previously that although both chemicals are immunogenic in mice, each provoking contact sensitization, exposure only to TMA results in an IgE antibody response. In the present study, to examine further the characteristics of human allergens, we have compared the ability of TMA and DNCB to elicit immediate and delayed cutaneous hypersensitivity reactions in mice. Topical exposure to both chemicals resulted in delayed (24 h) hypersensitivity. However, only TMA induced, in addition, an immediate (1 h) dermal reaction following local challenge. Serum from TMA-immune mice, but not from untreated mice or mice sensitized with DNCB, was able to transfer immediate hypersensitivity to naive recipients. The kinetics of passive sensitization with TMA-immune serum, together with the fact that immediate hypersensitivity to DNCB could be induced with monoclonal IgE anti-dinitrophenol (DNP) antibody, suggests that the immediate dermal responses caused by TMA are effected by hapten-specific IgE. These data demonstrate that different classes of occupational chemical allergen exhibit a variable potential to elicit immediate and delayed dermal hypersensitivity reactions in mice, and provide a novel approach to the classification and characterization of human allergens.

Administration, Topical↗
PubMed1992

Effects of ethanol on the decidual cell reaction in rats.

The effects of ethanol on uterine sensitivity to induction of decidualization and deciduoma growth were determined. Rats were ovariectomized, given an oestrogen-progesterone regimen to optimize induction and growth of deciduoma and randomly assigned to one of three ethanol treatment groups: (i) days 1-4 (pre-induction/period of sensitivity), (ii) days 5-9 (post-induction/period of growth), (iii) days 1-9 (periods of sensitivity and growth); or to a control group not treated with ethanol (pair-fed to treated groups). Ethanol (0, 1, 2, or 4 g kg-1) diluted in water was administered by stomach tube on the days prescribed. Decidualization was induced in one uterine horn by intraluminal injection of sodium phosphate buffer. Uterine sensitivity and decidual growth were assessed as cornu weight. Blood alcohol concentrations were measured by gas chromatography. Alcohol treatment reduced uterine sensitivity, but increased deciduoma growth. Blood alcohol concentrations rose to 133 mg% at 30 min, remained high for 90 min and declined to 82 mg% at 120 min. Thus, blood alcohol concentrations sufficient to induce mild intoxication in humans suppressed uterine sensitivity to decidualization and enhanced deciduoma growth in rats. As all ovarian steroid hormone support was exogenous, the effects of ethanol on deciduoma induction and growth were not due to alterations in the hypothalamic-pituitary-ovarian axis.

Animals↗
PubMed1992

Characterization of endothelin-converting enzyme from endothelial cells and rat brain: detection of the formation of biologically active endothelin-1 by rapid bioassay.

Using the endothelin-1 (ET-1)-stimulated elevation in cGMP in LLC-PK1 cells as a biological detector system for the conversion of big ET-1 (bET-1) to ET-1, we detected bET-1-converting activities in subcellular fractions from bovine aortic cultured endothelial cells (BAE) and rat brain. Within the particulate fraction of BAE, we detected two activities, at pH 3.4 and pH 5.4-7.4. The latter but not the former activity was inhibited in a concentration-dependent manner by phosphoramidon (approximate IC50, 1 microM) and converted bET-1 to ET-1 at a rate of 0.6 nmol/hr/mg of protein. It could be solubilized from the particulate fraction by detergent treatment. Phosphoramidon-inhibitable converting activity was also detected in the cytosolic fraction of BAE. Within the rat brain, phosphoramidon-inhibitable conversion of bET-1 to ET-1 was detected principally in the cytoskeletal fraction, i.e., that fraction from the membrane that was not solubilized by detergent treatment. These results show the presence of at least two different endothelin-converting enzyme activities in endothelial cells and a third within the rat brain. They also demonstrate the use of LLC-PK1 cells as a rapid assay that permits the sensitive detection and measurement of the formation of biologically active ET-1 from its precursor bET-1.

Animals↗
PubMed1992

Induction of NADPH-dependent diaphorase and nitric oxide synthase activity in aortic smooth muscle and cultured macrophages.

Lipopolysaccharide (LPS), either alone or in combination with cytokines, induces nitric oxide (NO) synthase activity in cells that normally release little or no NO. In arterial smooth muscle cells and various macrophage cell lines, NO synthase activity is induced after several hours of incubation with LPS. In brain, NADPH-dependent diaphorase activity has been associated with constitutive NO synthase. Here we show that incubation of rat aorta or cultured macrophages with LPS causes a time-dependent induction of NO synthase. The NO synthase activity in both rat aorta and macrophages was calcium independent and inhibited by NG-monomethyl-L-arginine and NG-nitro-L-arginine. We also found that LPS caused a time-dependent induction in NADPH-dependent diaphorase activity in both rat aorta and cultured macrophages. The diaphorase activity was mainly NADPH dependent and NADH independent. NO synthase activity and NADPH-diaphorase activity in crude cytosol from LPS-treated macrophages were found to co-purify, using 2',5'-ADP-Sepharose followed by Superose-6 gel permeation chromatography.

Amino Acid Oxidoreductases↗
PubMed1992

Preliminary evaluation of learning via the AI/LEARN/Rheumatology interactive videodisc system.

AI/LEARN/Rheumatology is a level three videodisc system to teach clinical observational skills in three important diseases: rheumatoid arthritis, osteoarthritis, and ankylosing spondylitis. The AI/LEARN software was developed on an independent authoring system called GALE designed for MS-DOS based computers. The purpose of this paper is to present preliminary data about the efficacy of teaching by the use of an interactive videodisc system as evaluated by examinations centered upon disease-oriented learning objectives and by attitude questionnaires. We tested the efficacy of the AI/LEARN/Rheumatology system using both medical students and residents taking the rheumatology elective. Data collected were on learning, attitudes, and ranking of curricular elements of the rotation. We kept records on the student time and search path through the interactive videodisc system. Control data were collected during 1990, before the AI/LEARN/Rheumatology program was available. Data for the treatment groups were collected during 1991 and 1992, while the trainees used the AI/LEARN/Rheumatology system. The basic difference between the control year and the treatment year curricula was the substitution of AI/LEARN/Rheumatology for three hours of lecture covering the three target diseases. AI/LEARN/Rheumatology was as effective as traditional methods of instruction as measured by scores on a multiple choice test. Student and resident learning was related to the time spent on the system. Students and residents ranked the AI/LEARN/Rheumatology system as the single most helpful learning tool in their 8 week rheumatology block, ranking it above the examination of patients.

Arthritis, Rheumatoid↗
PubMed1992

The Columbia Registry of Controlled Clinical Computer Trials.

Numerous reports on randomized controlled clinical trials of computer-based interventions have been published. These trials provide useful evaluations of the impact of information technology on patient care. Unfortunately, several obstacles make access to the trial reports difficult. Barriers include the large variety of publications in which reports may appear, non-standard descriptors, and incomplete indexing. Some analyzers indicate inadequate testing of computer methods. The purpose of establishing a registry of randomized controlled clinical computer trials was to assist the identification of computer services with demonstrated ability to improve the process or outcome of patient care. A report collection, selection, information extraction, and registration method was developed and implemented. One hundred and six reports on computer trials have been collected. A large variety of computer-assisted interventions have been tested in the registered trials (40% reminder, 15% feedback, 14% dose planning, 14% patient education, 12% medical record). 76% of the registered reports were published in the United States and most of the remainder in various European countries. In reporting computer trial results, 77% of the authors did not use both the "computer" and "trial" keywords in the title or abstract of their papers. We conclude that a major obstacle to adequate computer technology assessment is inadequate access to the published results.

Computer-Assisted Instruction↗
PubMed1991

Purification and characterization of particulate endothelium-derived relaxing factor synthase from cultured and native bovine aortic endothelial cells.

The particulate enzyme responsible for the synthesis of endothelium-derived relaxing factor has been purified from cultured and native (noncultured) bovine aortic endothelial cells. Purification of the solubilized particulate enzyme preparation by affinity chromatography on adenosine 2',5'-bisphosphate coupled to Sepharose followed by Superose 6 gel filtration chromatography resulted in a single protein band after denaturing polyacrylamide gel electrophoresis that corresponded to approximately 135 kDa. The enzyme activity in the various fractions was assayed by its stimulatory effect on soluble guanylyl cyclase of rat fetal lung fibroblasts (RFL-6 cells), by the formation of L-citrulline from L-arginine, by measuring nitrite/nitrate formation, and by bioassay on endothelium-denuded vascular strips. Endothelium-derived relaxing factor synthase was purified 3419-fold from the crude particulate fraction of cultured bovine aortic endothelial cells with a 12% recovery (RFL-6 assay). Purified endothelium-derived relaxing factor synthase required L-arginine, NADPH, Ca2+, calmodulin, and 5,6,7,8-tetrahydrobiopterin for full activity.

Amino Acid Oxidoreductases↗
PubMed1991

Endothelial cells have a particulate enzyme system responsible for EDRF formation: measurement by vascular relaxation.

Endothelium-derived relaxing factor (EDRF) released from endothelial cells (EC) has been shown to be nitric oxide (NO) or a closely related molecule. In cultured EC, the enzyme responsible for the formation of EDRF, EDRF-synthase, was initially described as being cytosolic, but more recently we have found it to be predominantly particulate. In view of this discrepancy we have investigated the EDRF synthesizing activity of cytosolic and particulate fractions isolated from native bovine aortic EC. EDRF was measured by cGMP formation in rat fetal lung cultured fibroblasts (RFL-6) and by the ability of cell fractions to relax endothelium-denuded, preconstricted rabbit aortic strips. Cytosolic fractions from native EC (100 micrograms) had no effect on the tone of rabbit aortic strips and little effect on cGMP levels in RFL-6 cells in the presence of L-arginine and NADPH (100 microM). However, under the same conditions the 100,000 x g pellet fractions relaxed rabbit aortic strips and increased cGMP levels in RFL-6 cells. Thus EDRF synthase from native EC, like those grown in culture, is located mainly in the particulate fraction.

Amino Acid Oxidoreductases↗
PubMed1991

A retinoic acid response element is part of a pleiotropic domain in the phosphoenolpyruvate carboxykinase gene.

Several hormones, including insulin, glucagon, and glucocorticoids, regulate the expression of the rate-limiting gluconeogenic enzyme, phosphoenolpyruvate carboxykinase [GTP: oxaloacetate carboxy-lyase (transphosphorylating); EC 4.1.1.32; PEPCK] in liver. In this report we demonstrate that retinoic acid (RA) also regulates PEPCK expression by inducing a 3-fold increase in the rate of transcription of the PEPCK gene. A RA response element located between -468 and -431 in the PEPCK promoter mediates a 7-fold increase in expression of a chimeric construct containing the basal PEPCK promoter ligated to the chloramphenicol acetyltransferase reporter gene. This element confers RA responsiveness through the heterologous thymidine kinase promoter and functions relatively independent of position and orientation. An 18-base-pair core sequence (-451 to -434) (i) mediates an effect of RA on PEPCK gene expression and contains motifs found in two other RA response elements; (ii) corresponds to AF1, an accessory factor element that is an integral component of the complex glucocorticoid response unit in the PEPCK gene promoter; (iii) is in a region involved in the developmental expression of the PEPCK gene; and (iv) shows homology to elements involved in the tissue-specific regulation of genes, including the hepatic apolipoprotein genes and the alpha 1-antitrypsin gene.

Animals↗
PubMed1991

The Medline/full-text research project.

This project was designed to test the relative efficacy of index terms and full-text for the retrieval of documents in those MEDLINE journals for which full-text searching was also available. The full-text files used were MEDIS from Mead Data Central and CCML from BRS Information Technologies. One hundred clinical medical topics were searched in these two files as well as the MEDLINE file to accumulate the necessary data. It was found that full-text identified significantly more relevant articles than did the indexed file, MEDLINE. The full-text searches, however, lacked the precision of searches done in the indexed file. Most relevant items missed in the full-text files, but identified in MEDLINE, were missed because the searcher failed to account for some aspect of natural language, used a logical or positional operator that was too restrictive, or included a concept which was implied, but not expressed in the natural language. Very few of the unique relevant full-text citations would have been retrieved by title or abstract alone. Finally, as of July, 1990 the more current issue of a journal was just as likely to appear in MEDLINE as in one of the full-text files.

Abstracting and Indexing↗
PubMed1991

Endometrial arylamidase activity in the guinea pig: changes during the oestrous cycle, decidualization and ovarian steroid hormone treatment.

1. As suggested by comparative studies done in various species, amino acid arylamidases (amino peptidases) may play a role in blastocyst implantation. 2. Histochemical studies of the guinea pig endometrium indicate that arylamidase increases in the stroma during pregnancy but is depleted in the vicinity of the blastocyst during implantation. 3. To further explore the possible significance of arylamidases in uterine function, endometrial arylamidase activity was measured in guinea pigs during the reproductive cycle, decidualization and after ovariectomy with and without estrogen (E) and/or progesterone (P) treatment. Arylamidase activity was maximal during pro-oestrus-oestrus (40.0 +/- 10.0 mu/mg protein). 4. Enzyme activity was markedly depleted in decidualized endometrial stroma (12.3 +/- 1.6, P less than 0.01); reduced by ovariectomy (20.5 +/- 2.7); and stimulated by E (29.2 +/- 1.2); P had little effect (21.9 +/- 3.5). 5. The physiological significance of modulation of endometrial arylamidase activity by steroid hormones is discussed.

Aminopeptidases↗
PubMed1991

Alkaline buffers release EDRF from bovine cultured aortic endothelial cells.

1. Release of endothelium-derived relaxing factor (EDRF) and prostacyclin (PGI2) from bovine cultured aortic endothelial cells (EC) was measured by bioassay and radioimmunoassay, respectively. 2. Bradykinin (BK, 3-30 pmol), adenosine diphosphate (ADP, 2-6 nmol) or the sodium ionophore monensin (40-100 nmol) injected through a column of EC released EDRF. L-Arginine free base (FB; 10-20 mumol) or D-arginine FB (10-20 mumol) injected through the column of EC released similar amounts of EDRF and also caused an increase in pH of the Krebs solution perfusing the EC from 7.5-8.0 to 8.6-9.5. Sodium carbonate (Na2CO3) an alkaline buffer which caused the same changes in the pH of the Krebs solution also induced the same release of EDRF. The hydrochloride salts of L- or D-arginine did not cause either release of EDRF when injected through the column of EC or increases in the pH of the Krebs solution. 3. Inhibitors of either diacylglycerol lipase (RHC 80267) or kinase (R59022) inhibited the release of EDRF induced by BK or ADP but potentiated the release induced by L-arginine FB, monensin (40-100 nmol) or alkaline buffer (Na2CO3). R59022 and RHC 80267 infused through the EC increased the basal release of EDRF. 4. When calcium chloride was omitted from the Krebs solution the release of EDRF induced by alkaline buffer (Na2CO3; pH 8.6-9.5) or L-arginine FB (10-20 mumol) was selectively inhibited when compared to that induced by BK (3-30 pmol) or ADP (2-6 nmol). This inhibition was reversed when calcium (2.5 mM) was restored. 5. NG-monomethyl-L-arginine (NMMA; 30 microM) inhibited release of EDRF induced by BK (10-30 pmol) or alkaline buffers (Na2CO3 or D-arginine FB; pH 8.6-9.5). This inhibition was partially reversed by L- but not D-arginine FB or HCl (30-100 microM). 6. Prostacyclin was released when BK (10 pmol), ADP (2 nmol) or arachidonic acid (30 nmol) were injected through the column of EC. However, monensin (40 nmol) or alkaline buffers (pH 8.6-9.5) did not release detectable amounts of PGI2 as measured by radioimmunoassay for 6-oxo-prostaglandin F1 alpha. 7. Thus alkalinisation of the external bathing solution can release EDRF from cultured EC by a mechanism which does not involve receptor activation and which depends on the presence of extracellular calcium.

Animals↗
PubMed1991

Inhibition of the release of endothelium-derived relaxing factor in vitro and in vivo by dipeptides containing NG-nitro-L-arginine.

1. We have shown that dipeptides containing NG-nitro-L-arginine (NO2Arg) inhibit the biosynthesis of endothelium-derived relaxing factor (EDRF) in vitro and in vivo. 2. In anaesthetized rats, intravenous administration at 1-30 mg kg-1 of the methyl ester of NO2Arg, NO2-Arg-L-phenylalanine (NO2Arg-Phe), L-alanyl-NO2Arg (Ala-NO2Arg) or NO2Arg-L-arginine (NO2Arg-Arg) produced dose-related increases in mean arterial blood pressure (MABP) which were unaffected by D-arginine (D-Arg; 20 mg kg-1 min-1 for 15 min), but prevented by co-infusions of L-arginine (L-Arg; 20 mg kg-1 min-1 for 15 min) or by their parent dipeptides. 3. NO2Arg methyl ester, NO2Arg-Phe methyl ester or Ala-NO2Arg methyl ester (10 mg kg-1, i.v.) also inhibited the reduction in MABP caused by the endothelium-dependent vasodilator, acetylcholine (30 micrograms kg-1 min-1 for 3 min), but not those induced by glycerly trinitrate (20 micrograms kg-1 min-1 for 3 min) or iloprost (6 micrograms kg-1 min-1 for 3 min) which act directly on the vascular smooth muscle. 4. Moreover, NO2Arg methyl ester, NO2Arg-Phe methyl ester or NO2Arg-Arg methyl ester (100 microM) inhibited the acetylcholine-induced relaxation of rabbit aortic strips, and NO2Arg-Phe methyl ester (30 microM) blocked the stimulated (bradykinin, 30 pmol) release of EDRF from bovine aortic endothelial cells grown on microcarrier beads. 5. In endothelial cells grown in L-Arg-deficient medium, L-Arg-containing dipeptides such as L-Arg-LPhe, L-Ala-L-Arg or L-Arg-L-Arg increased both the basal and stimulated release of EDRF. Moreover, the L-Arg containing dipeptides, but not their NO2Arg analogues, were rapidly cleaved by these cells. 6. Thus, dipeptides containing NO2Arg can directly interfere with the biosynthesis of EDRF in vitro and in vivo. Moreover, the potentiation of EDRF release from endothelial cells deprived of L-Arg by dipeptides containing L-Arg suggests that such peptides may serve as an additional or alternative substrate for the biosynthesis of EDRF.

Amino Acid Sequence↗