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Biomedical subjects

W Burgess

Publications and source records attributed to W Burgess.

At least 37 records · Page 2Linked to original sources
PubMed1988

Regulation of plasminogen activator inhibitor-1 mRNA in human endothelial cells.

The plasminogen activator inhibitor (PAI-1) from endothelial cells is a potentially important regulator of plasminogen activator activity. Cultured human endothelial cells increase their PAI-1 production upon stimulation with LPS and TNF, agents that are known to cause an increase in PAI-1 levels in vivo. We isolated a PAI-1 cDNA probe, and by RNA hybridization analysis studied the regulation of PAI-1 mRNA synthesis in human endothelial artery cells. Freshly isolated endothelial cells do not contain detectable amounts of PAI-1 mRNA, but after adherence and incubation for 18 h in growth medium produce considerable amounts of PAI-1 activity and contain PAI-1 mRNA levels comparable to those found in subcultured cells. When subcultured endothelial cells are incubated for 6 h with LPS or TNF, both species of PAI-1 mRNA increase 10 to 20 fold, while PAI-1 activity in the growth medium increases only 1.5 to 2 fold. Stimulation of endothelial cells in the presence of cycloheximide (CHX) results in superinduction of mainly the 3.0 kb PAI-1 mRNA. The 3' end of this mRNA contains a 60 bp AT-rich sequence, that resembles 3' sequences present in a number of other genes superinducible with CHX.

Base Sequence↗
PubMed1988

Specificity of rabbit cytochrome P-450 isozymes involved in the metabolic activation of the food derived mutagen 2-amino-3-methylimidazo[4,5-f] quinoline.

The involvement of rabbit liver cytochromes P-450 in the activation of the food derived heterocyclic amine mutagen, 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), was assessed using the Ames/Salmonella test. The number of revertants induced by IQ per microgram of control rabbit liver microsomes was 1872 +/- 50 (SD, n = 3), and this increased to 3690 +/- 239 when microsomes from 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) animals were used as the metabolic activation source. Microsomes from phenobarbital, rifampicin, and acetone pretreated rabbits were less efficient the controls at activating IQ to a mutagen. Cytochrome P-450 Forms 4 and 6, which are induced by TCDD, were found to be efficient activators of IQ to a mutagen in reconstitution experiments. Form 4 was 7.7-fold more active than Form 6 and produced 1702 revertants/0.125 pmol with a 20-min preincubation step in the Ames test. Anti-Form 4 IgG inhibited the activation of IQ in control and TCDD induced microsomes by 78 and 79%, respectively. The contents of cytochrome P-450 Form 4, determined by Western blot analysis, in control and phenobarbital, acetone, rifampicin, and TCDD pretreated microsomes were 0.55 +/- 0.19, 0.63 +/- 0.34, 0.5 +/- 0.27, 0.28 +/- 0.16, and 2.19 +/- 0.43 (n = 3) nmol/mg protein, respectively. A highly significant statistical correlation existed between the capacity of the above microsomes to activate IQ to a mutagen and their cytochrome P-450 Form 4 content (r = 0.96; r2 = 0.92). The content of cytochrome P-450 Form 6 in the above microsomes was also highly correlated with their capacity to activate IQ (r = 0.92; r2 = 0.85). Based on these results and the tissue distribution of cytochrome P-450 Forms 4 and 6, the former obviously contributes most toward the activation of IQ in the liver, whereas Form 6 would be expected to be primarily involved in this process in extrahepatic tissues.

Animals↗
PubMed1988

Activation of the food-derived mutagen 2-amino-3-methylimidazo[4, 5-f]quinoline by human-liver microsomes.

The ability of human-liver microsomes to metabolically activate the food-derived heterocyclic amine, 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), and the model mutagen, 2-aminofluorene (AF), has been investigated using Salmonella typhimurium TA98. In 6 subjects tested the number of revertants produced by 0.1 micrograms IQ per mg microsomal protein varied from 11, 830 +/- 320 to 42, 830 +/- 290 (mean +/- SD). With the same livers and a dose of 10 micrograms AF per plate the number of revertants varied from 15,770 +/- 1600 to 29,380 +/- 810 per mg microsomal protein. Metyrapone and alpha-naphthoflavone caused differential inhibition of the mutagenesis of both IQ and AF indicating the involvement of different forms of cytochrome P450 in the metabolic activation of these amines in human-liver microsomes. In presence of human-liver microsomes IQ produced no detectable increase in mutations at the hypoxanthine phosphoribosyl transferase locus in lymphocytes and caused no increase in micronuclei formation at realistic exposure levels.

Benzoflavones↗
PubMed1988

Foot impact force and intravascular hemolysis during distance running.

This study was designed to examine the relationship between foot impact force and the magnitudes of the changes in markers of intravascular hemolysis during distance running. Fourteen male distance runners (VO2 max = 66.1 +/- 5.0 ml.kg-1.min-1, mean +/- SD) completed two treadmill runs and a resting control procedure. The two treadmill tests involved running at 215 m.min-1 for 10,000 footstrikes at elevations of either +6% (uphill) or -6% (downhill). Mean foot impact force was 11% greater with the downhill than the uphill running. The three procedures were ordered randomly and separated by 7 days. Hemoglobin concentration (Hb), hematocrit (Hct), plasma free hemoglobin (PFHb), and haptoglobin (Hp) concentrations were assayed in blood samples collected via venipuncture 15 min and immediately before exercise, and immediately, 1 h, and 2 h after the exercise. Repeated measures ANOVA revealed that Hp was significantly decreased and PFHb was significantly increased after treadmill running (P less than 0.05) and that these changes were significantly greater with downhill than uphill running (P less than 0.05). These findings support the theory that mechanical trauma to red blood cells occurring at footstrike is a major cause of hemolysis during running.

Adult↗
PubMed1988

Identification and quantitation in human liver of cytochromes P-450 analogous to rabbit cytochromes P-450 forms 4 and 6.

1. Polyclonal antibodies raised against rabbit liver cytochrome P-450 isozymes form 4 and 6 have been used to probe human liver microsomes for analogous proteins using the Western blot technique. 2. Anti-Form 4 IgG recognized a protein in human liver microsomes from six subjects of identical molecular weight to purified rabbit liver cytochrome P-450 Form 4. 3. The equivalent content of cytochrome P-450 Form 4 in the same microsomes ranged from 1.1 to 9.1 pmol per mg protein. 4. Anti-Form 6 IgG recognized a protein in human liver microsomes from the same six subjects of slightly higher molecular weight than purified rabbit cytochrome P-450 Form 6. 5. The equivalent content of cytochrome P-450 Form 6 in the above microsomes ranged from 1.6 to 3.8 pmol per mg protein. 6. No significant correlations were observed between equivalent cytochrome P-450 Forms 4 and 6 content and 2-acetylaminofluorene N-hydroxylase, aminopyrine N-demethylase, benzyprene and aniline hydroxylase activities in liver microsomes from the six subjects tested.

Adolescent↗
PubMed1987

Effect of dexamethasone on cytochrome P-450 mediated metabolism of 2-acetylaminofluorene in cultured rat hepatocytes.

The metabolism of 2-acetylaminofluorene (AAF) to its six oxidative metabolites has been used to investigate the effect of dexamethasone on cytochrome P-450 activity in cultured rat hepatocytes. In control hepatocytes the metabolism of AAF to its 1-, 5-, 7-, 9- and N-hydroxylated metabolites rapidly declined in culture over the first 24 hr while 3-hydroxylation remained relatively constant. These activities either remained unchanged or increased slightly during the next 48 hr in culture. The addition of dexamethasone (100 nM) to the culture medium had little effect in arresting the initial decline but by 72 hr the 7-, 5- and 3-hydroxylations increased to values 2.5, 16 and 21 times the respective 24-hr values. The inductive effect of dexamethasone on the 3- and 5-hydroxylations of AAF was maximal at 100 nM whereas the 7-hydroxylation increased linearly as a function of the dexamethasone concentration up to 1 microM. Cortisol and corticosterone and the non-glucocorticoids fluoxymesterone and methyltestosterone induced a pattern of AAF metabolism resembling that in dexamethasone-treated cultures, suggesting that a range of steroids not restricted to glucocorticoids may induce multiple cytochrome P-450 isozymes via related mechanisms. Pregnenolone 16 alpha-carbonitrile induced only the 7-hydroxylation of AAF probably reflecting induction of cytochrome P-450p. While dexamethasone was a strong inducer of the 3- and 5-hydroxylations of AAF in hepatocyte culture, assay of these activities in freshly isolated cells after in vivo treatment with dexamethasone showed a strong induction of 7-hydroxylation but only small effects on 3- and 5-hydroxylations. Indeed the profile of AAF metabolism induced in culture by dexamethasone resembles more closely the profile induced by 3-methylcholanthrene in vivo. These data suggest that factors yet to be identified strongly influence the steroid-induced pattern of cytochrome P-450 gene expression.

2-Acetylaminofluorene↗
PubMed1987

Immunohistochemical localization and quantitation of NADPH-cytochrome P-450 reductase in human liver.

An antibody raised in a goat against the human liver NADPH-cytochrome P-450 reductase (EC 1.6.2.4.) enzyme has been used to: 1) immunoquantify the level of this enzyme in human liver microsomes, and 2) study the distribution of the reductase across the human liver acinus. Employing the Western blot procedure, anti-human reductase IgG recognized a single band in human liver microsomes which corresponded in molecular weight to the purified reductase. The content of the NADPH-cytochrome P-450 reductase in six normal human livers varied from 87 to 121 pmol/mg of microsomal protein. NADPH-cytochrome P-450 reductase activity of the same microsomes ranged from 107 to 222 nmol of cytochrome c reduced per min per mg of protein. The correlation between reductase content and activity (r = 0.54) was not statistically significant (p greater than 0.1). The total cytochrome P-450 content (cytochrome P-450 and P-420) of the same microsomes varied from 423 to 1413 pmol/mg of microsomal protein. The average ratio of cytochrome P-450 to NADPH-cytochrome P-450 reductase was 7.1:1 +/- 3.1 (mean +/- SD) in the human liver microsomal preparations studied. The reductase was found to be nonuniformly distributed across the human liver acinus. Although all hepatocytes stained positively for NADPH-cytochrome P-450 reductase, the staining intensity was highest in zone 3 and in some cases also in zone 1 hepatocytes. These results show that human liver contains a gross excess of cytochrome P-450 molecules to NADPH-cytochrome P-450 reductase molecules. Furthermore, the differential distribution of the reductase within the human liver acinus may lead to a better understanding of the mechanism underlining site-specific drug hepatotoxicity.

Cytochrome P-450 Enzyme System↗
PubMed1987

Metabolism of 2-acetylaminofluorene in cultured human lymphocytes.

The capacity of lymphocytes from 23 human subjects to metabolize the model carcinogen 2-acetylaminofluorene (AAF) was assessed. These cells metabolized AAF to its 1-, 7- and N-hydroxylated metabolites. Alpha-naphthoflavone and metyrapone exhibited IC50's for all pathways of 0.02 microM and 1 mM, respectively. These data indicate that the same form of cytochrome P450 or forms with similar IC50's are mediating these reactions. No significant difference was observed between lymphocytes from smokers versus non-smokers in the N-hydroxylation of AAF, the initial step in the metabolic activation of this carcinogen. However, lymphocytes from smokers were faster detoxifiers of AAF as seen in their increased capacity to 1- and 7-hydroxylate AAF compared to lymphocytes from non-smokers.

2-Acetylaminofluorene↗
PubMed1986

Human endothelial cell growth factor: cloning, nucleotide sequence, and chromosome localization.

Several of the endothelial cell polypeptide mitogens that have been described probably play a role in blood vessel homeostasis. Two overlapping complementary DNA clones encoding human endothelial cell growth factor (ECGF) were isolated from a human brain stem complementary DNA library. Southern blot analysis suggested that there is a single copy of the ECGF gene and that it maps to human chromosome 5 at bands 5q31.3 to 33.2 A 4.8-kilobase messenger RNA was present in human brain stem messenger RNA. The complete amino acid sequence of human ECGF was deduced from the nucleic acid sequence of these clones; it encompasses all the well-characterized acidic endothelial cell polypeptide mitogens described by several laboratories. The ECGF-encoding open reading frame is flanked by translation stop codons and provides no signal peptide or internal hydrophobic domain for the secretion of ECGF. This property is shared by human interleukin-1, which is approximately 30 percent homologous to ECGF.

Base Sequence↗
PubMed1977

Rape: power, anger, and sexuality.

Accounts from both offenders and victims of what occurs during a rape suggest that issues of power, anger, and sexuality are important in understanding the rapist's behavior. All three issues seem to operate in every rape, but the proportion varies and one issue seems to dominate in each instance. The authors ranked accounts from 133 offenders and 92 victims for the dominant issue and found that the offenses could be categorized as power rape (sexuality used primarily to express power) or anger rape (use of sexuality to express anger). There were no rapes in which sex was the dominant issue; sexuality was always in the service of other, nonsexual needs.

Adult↗
PubMed1974

Outbreak of meningococcal disease in Devon.

In a recent outbreak of 31 cases of meningococcal disease in Devon there were six deaths. Several patients had an unusual rash as the presenting feature and there was an unusually high incidence of complications, affecting the central nervous system, joints, and the heart among other sites.

Adolescent↗