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Biomedical subjects

M Mushtaq

Publications and source records attributed to M Mushtaq.

At least 19 recordsLinked to original sources

Steatosis and chronic hepatitis C virus infection.

OBJECTIVE: To asses the presence of steatosis and other histological changes in patients with chronic hepatitis C. METHODS: Liver biopsy samples were reviewed for presence of steatosis, its degree and other histological changes of hepatitis C including necro-inflammatory score, fibrosis grade and these changes were correlated with liver function tests. RESULTS: A total of 109 liver biopsy samples were reviewed. Mean age of the patients was 44.46 +/- 13.93 years and 62 (56.8%) were male. Mean necro-inflammatory score was 2.32 +/- 0.95. Mean fibrosis grade was 1.69 +/- 1.12. No Steatosis was found in 42 (38.5%) samples and mild to severe degree of steatosis was found in 67 (61.5%) samples. CONCLUSION: In this study, nearly 62% of liver biopsy samples had some degree of steatosis. Whether this steatosis increases occurrence of severe form of fibrosis or is an associated phenomena with chronic inflammation needs to be further clarified.

Adult↗

Early switch from intravenous to oral antibiotics and early hospital discharge: a prospective observational study of 200 consecutive patients with community-acquired pneumonia.

UNLABELLED: To determine the proportion of patients who can be treated with early switch to oral antibiotics and early discharge, to evaluate clinical outcome and patient satisfaction for patients treated with early switch and early discharge, and to define the factors that interfere with early discharge for some of the patients who underwent early switch to oral antibiotic therapy. DESIGN: Prospective study. PARTICIPANTS: Two hundred consecutive hospitalized patients with community-acquired pneumonia. MAIN OUTCOME MEASURES: Number of days needed to switch to oral therapy and length of hospital stay. Clinical outcome and satisfaction with care were evaluated for those patients treated with early switch and early discharge. RESULTS: Early switch to oral antibiotics (within the first 3 days of hospitalization) was performed in 133 patients (67%). Clinical failure was documented in 1 patient. Early switch and early discharge was performed in 88 patients (44%). The mean length of hospital stay for this group was 3.4 days. The most common reason for prolonged hospitalization after the switch to oral antibiotics was the need for diagnostic workup. More than 95% of patients were satisfied with the care they had received. CONCLUSIONS: Using simple clinical and laboratory criteria, a significant proportion of hospitalized patients with community-acquired pneumonia (44%) can be treated with early switch and early discharge. This model did not affect patient outcome, decreased the length of hospitalization, and was associated with a high level of patient satisfaction.

Administration, Oral↗

A re-audit of pulmonary function laboratories in the West Midlands.

In 1991 the West Midlands Pulmonary Function Audit Group examined the consistency between pulmonary function laboratories in the West Midlands. Three healthy subjects visited 22 centres and performed a standard set of pulmonary function tests. Demographic data on nine hypothetical subjects was also supplied for the laboratories to produce predicted values. Equipment was checked for accuracy using standard methods. The 1991 audit revealed significant inter-laboratory variability. Sources of error were identified and after consultation, recommendations were made to improve consistency. In addition, national and regional training workshops were organized for laboratory staff. In 1995 the audit was repeated using the same three subjects. Significant differences continued for all predicted results except for residual volume (RV) and forced vital capacity (FVC) and for all measured results except for functional residual capacity (FRC). However, improvements in the coefficient of variation were seen compared with 1991 for predicted forced expiratory volume (FEV1), total lung capacity (TLC), gas transfer (TLCO), FVC, FRC and RV. Similar improvements were seen in measured results for FEV1 and FVC. Increased variation was seen for predicted corrected transfer factor (KCO) and actual RV. The majority of variables in the 1995 audit had a coefficient of variation of less than 5% between laboratories. Analysis of the predicted results from the hypothetical subjects continued to show unacceptable variation reflecting continuing computer algorithm inconsistency. The improvements seen are encouraging and suggest that a regular audit programme is worthwhile.

Adult↗

Metabolism of 3H/14C-labeled 4''-deoxy-4''-epimethylaminoavermectin B1a benzoate in chickens. Identification of novel fatty acid conjugates of '4'-deoxy-4''-epimethylaminoavermectin B1a.

The metabolism of 3H/14C-labeled 4"-deoxy-4"-epimethylaminoavermectin B1a (MAB1a) benzoate, the major homologue (>/=90%) of the avermectin insecticide emamectin benzoate, was studied in laying chickens. Ten Leghorn hens (Gallus domesticus) were orally dosed once daily for 7 days (1 mg/kg of body weight/day). Eggs and excreta were collected daily, and eggs were subsequently separated into whites and yolks. Chickens were euthanized within 20 hr after the last dose, and liver, kidney, heart, muscle, fat, ovaries, gizzard, gastrointestinal tract and contents, and carcass were collected. Approximately 70 and 6% of the total administered dose were recovered in the excreta plus gastrointestinal tract and contents and in the tissues plus eggs, respectively. Two novel metabolites, i.e. the 24-hydroxymethyl derivative of the parent compound (24-hydroxymethyl-4"-deoxy-4"-epimethylaminoavermectin B1a) and the N-demethylated derivative of 24-hydroxymethyl-4"-deoxy-4"-epimethylaminoavermectin B1a (24-hydroxymethyl-4"-deoxy-4"-epiaminoavermectin B1a), were identified. In addition, eight fatty acid conjugates of each of these two metabolites, comprising 8-75% of total radioactive residues in tissues and eggs, were isolated and identified. Although this represents some of the most extensive in vivo fatty acid conjugation to a xenobiotic reported to date, potential human exposure to MAB1a residues from consumption of chicken would be extremely low, because the dosage level in this study was approximately 1000-fold greater than the MAB1a residue levels seen in crops and because the majority of the applied dose was recovered in the excreta. Based on these findings, the avian biotransformation of MAB1a differs substantially from the mammalian biotransformation.

Animals↗

Gossypol inhibits human chorionic gonadotropin-stimulated testosterone production by cultured canine testicular interstitial cells.

Gossypol (GP) is a natural polyphenolic compound that possesses antifertility and antisteroidogenic activities in both males and females. The dog is highly sensitive to GP toxicity, yet GP's effect on canine testicular steroidogenesis has never been reported. Thus, the present study examines GP's effects on human chorionic gonadotropin (hCG)-induced testosterone (T) production by primary cultured canine testicular interstitial cells. After decapsulation and enzymatic dissociation of canine testes in Dulbecco's Modified Eagle Medium with Ham's Nutrient Mixture F-12 (1:1; DME/F-12) containing 0.1% collagenase, 0.1% BSA, and 10 micrograms/ml DNase 1 (37 degrees C, 20 min), interstitial cells were isolated by sedimentation and filtration (140 microns) and then cultured in supplemented DME/F-12 medium (5 micrograms/ml insulin, 5 micrograms/ml transferrin, 5 ng/ml sodium selenite; DME/F-12/S) containing 0.1% fetal bovine serum (FBS). FBS was used to enhance cell attachment during the first 24 hours of culture. After 24 hours, the medium was replaced with serum-free DME/F-12/S and the cells were cultured for an additional 24 hours. Thereafter, cells were treated with hCG (0.1 IU/ml) alone and in combination with GP (0.05, 0.5, 2.5 and 5.0 microM). Media were collected for T radioimmunoassay and cells for protein estimation after 8, 16 and 24 hours of treatment. Treatment with hCG significantly (p < 0.05) stimulated T production over that of controls at all treatment times examined. At 8, 16 and 24 hours, T secretion was elevated from 0.91 +/- 0.25, 1.32 +/- 0.42, and 1.41 +/- 0.40 pg/microgram protein to 2.36 +/- 0.50, 2.84 +/- 0.60, and 2.82 +/- 0.43 pg/microgram protein, respectively. At 0.5, 2.5 and 5.0 microM, GP significantly (p < 0.05) reduced hCG-induced T secretion at 16 and 24 hours of treatment to 1.79 +/- 0.50, 1.62 +/- 0.12, 1.34 +/- 0.16 (16 hr), and 1.53 +/- 0.38, 1.43 +/- 0.11, 1.42 +/- 0.32 (24 hr) pg/microgram protein, respectively. At 8 hours, T production was reduced by 2.5 and 5.0 microM GP to 1.08 +/- 0.55 and 0.93 +/- 0.61 pg/microgram protein, respectively. GP, however, did not reduce T production to below basal levels. These results demonstrate the inhibition of hCG-induced T production by GP in cultured canine testicular interstitial cells.

Animals↗

An audit of pulmonary function laboratories in the West Midlands.

BACKGROUND: Pulmonary function testing has become an integral part of the assessment and follow-up of patients with pulmonary disease. Many factors can influence the results produced by a laboratory. This audit was performed to examine the extent of variation in the pulmonary function test results amongst all laboratories in the West Midlands. This was followed by an attempt to determine the cause of this variation. METHODS: Phase 1. Three normal healthy subjects each underwent a set of pulmonary function tests in all 22 laboratories in the West Midlands. Information regarding technicians' qualifications, training and seniority, protocol and equipment used were obtained in the form of a questionnaire. Phase 2. All 22 laboratories were asked to calculate the predicted values on the same nine sets of demographic data. These data included both sexes, ethnic minorities and range of ages. In addition technical aspects of each laboratory were investigated including the assessment of volume and gas analysers with standard gases containing known concentrations of helium and carbon monoxide. RESULTS: Phase 1. Significant variations (P < 0.05) were observed in all measured values of pulmonary function tests of the three subjects. Significant variations (P < 0.05) were also observed in all predicted values except total lung capacity. Phase 2. There were significant variations (P < 0.05) amongst laboratories in calculating the predicted values of all components of pulmonary function tests. No significant differences were observed in the measurement of volume or concentration of carbon monoxide or helium.

Adult↗

Multiple cystic pulmonary hamartomas.

A patient with multiple cystic hamartomas presented with a pneumothorax and later developed a cystic myxomatous vaginal polyp. This and three of the cysts were resected. She remains well 13 years later. Multiple cystic hamartomas are uncommon and may be misdiagnosed as pulmonary metastases.

Cysts↗

Absolute configuration of cis-5,6-dihydrodiol enantiomers derived from helical conformers of 1,12-dimethylbenz[a]anthracene.

1,12-Dimethylbenz[a]anthracene (1,12-DMBA) cis-5,6-dihydrodiol was synthesized by oxidation of 1,12-DMBA with osmium tetroxide in pyridine in low yield (less than or equal to 3%) and was purified by sequential use of reversed-phase and normal-phase HPLC. Two pairs of 1,12-DMBA cis-5,6-dihydrodiol enantiomers, derived from P (right-handed helix) and M (left-handed helix) conformers, were eluted as a single chromatographic peak on both reversed-phase and normal-phase HPLC. However, these four enantiomers were resolved by sequential use of two chiral stationary phase (CSP) HPLC columns. CSP (Pirkle type I) columns were packed with either (R)-N-(3,5-dinitrobenzoyl)phenylglycine or (S)-N-(3,5-dinitrobenzoyl)leucine, which is ionically bonded to gamma-aminopropylsilanized silica. Absolute configurations of enantiomers were determined by comparing their circular dichroism spectra with those of conformationally similar cis-5,6-dihydrodiol enantiomers of 4-methylbenz[a]anthracene and 7,12-dimethylbenz[a]anthracene with known absolute stereochemistry.

9,10-Dimethyl-1,2-benzanthracene↗

Improved enantiomeric separation of dihydrodiols of polycyclic aromatic hydrocarbons on chiral stationary phases by derivatization to O-methyl ethers.

K-region trans-dihydrodiol derivatives of phenanthrene, 1-methylphenanthrene, 4,5-methylenephenanthrene, pyrene, 1-bromopyrene, chrysene, benzo[c]phenanthrene, benz[a]anthracene, 1-, 4-, 6-, 7-, 11- and 12-methylbenz[a]anthracenes, 7,12-dimethylbenz[a]anthracene, 3-methylcholanthrene, and benzo[a]pyrene, and non-K-region trans-3,4-dihydrodiols of benz[a]anthracene, chrysene, and 7,12-dimethylbenz[a]anthracene are converted to O-methyl ethers. Enantiomers of these O-methyl ethers are generally more efficiently separated on Pirkle's chiral stationary phases than the enantiomers of underivatized dihydrodiols. O-Methyl ethers are substantially less polar than dihydrodiols, and O-methyl ethers are eluted with shorter retention times. Eluents of lower polarity can hence be used. This enhances chiral interactions between chiral stationary phase and solutes, allowing improved separation of enantiomers.

Chemical Phenomena↗

Stereoselective formation and hydration of 12-methylbenz[a]anthracene 5,6-epoxide enantiomers by rat liver microsomal enzymes.

The K-region trans-5,6-dihydrodiols formed in the metabolism of 12-methylbenz[a]anthracene (12-MBA) by liver microsomal preparations from untreated, phenobarbital-treated and 3-methylcholanthrene-treated male Sprague-Dawley rats were found by chiral stationary-phase h.p.l.c. (c.s.p.-h.p.l.c.) analyses to contain (5S,6S)/(5R,6R) enantiomer ratios of 93:7, 88:12 and 97:3 respectively. The absolute stereochemistry of a 12-MBA trans-5,6-dihydrodiol enantiomer was elucidated by the exciton-chirality c.d. method. The 5,6-epoxides formed in the metabolism of 12-MBA by liver microsomal preparations from untreated, phenobarbital-treated and 3-methylcholanthrene-treated male Sprague-Dawley rats in the presence of the epoxide hydrolase inhibitor 3,3,3-trichloropropylene 1,2-oxide were isolated from a mixture of metabolites by normal-phase h.p.l.c., and their (5S,6R)/(5R,6S) enantiomer ratios were found by c.s.p.-h.p.l.c. analyses to be 73:27, 78:22 and 99:1 respectively. The absolute configurations of 12-MBA 5,6-epoxide enantiomers, resolved by c.s.p.-h.p.l.c., were determined via high-resolution (500 MHz) proton-n.m.r. and c.d. spectral analyses of the two isomeric methoxylation products derived from each of the 12-MBA 5,6-epoxide enantiomers. Enantiomeric pairs of the two methoxylation products were resolved by c.s.p.-h.p.l.c. The results indicate that enantiomeric 5S,6R-epoxide and 5S,6S-dihydrodiol were the major enantiomers preferentially formed in the metabolism at the K-region 5,6-double bond of 12-MBA by all three rat liver microsomal preparations. Optically pure 12-MBA 5S,6R-epoxide was hydrated predominantly at the C(6) position (R centre) to form 12-MBA trans-5,6-dihydrodiol with a (5S,6S)/(5R,6R) enantiomer ratio of 97:3. However, optically pure 12-MBA 5R,6S-epoxide was hydrated nearly equally at both C(5) and C(6) positions to form 12-MBA trans-5,6-dihydrodiol with a (5S,6S)/(5R,6R) enantiomer ratio of 57:43.

Animals↗

Stereoselective formation and hydration of benzo[c]phenanthrene 3,4- and 5,6-epoxide enantiomers by rat liver microsomal enzymes.

The K-region 5,6-epoxides, formed in the metabolism of benzo[c]phenanthrene (BcPh) in the presence of an epoxide hydrolase inhibitor 3,3,3-trichloropropylene 1,2-oxide (TCPO) by liver microsomes from untreated, phenobarbital-treated, 3-methylcholanthrene-treated, and polychlorinated biphenyls (Aroclor 1254)-treated rats of the Sprague-Dawley and the Long-Evans strains, were found by chiral stationary phase high-performance liquid chromatography analyses to be enriched (58-72%) in the 5S, 6R enantiomer. In the absence of TCPO, the metabolically formed BcPh trans-5,6-dihydrodiol was enriched (78-86%) in the 5S,6S enantiomer. The major enantiomer of the BcPh 3,4-epoxide metabolite was found to be enriched in the 3S,4R enantiomer which undergoes racemization under the experimental conditions. The major enantiomer of the 5,6-dihydrodiol metabolite was elucidated by the exciton chirality circular dichroism (CD) method to have a 5S,6S absolute stereochemistry. Absolute configurations of enantiomeric methoxylation products derived from each of the two BcPh 5,6-epoxide enantiomers. Optically pure BcPh 5S,6R-epoxide was enzymatically hydrated exclusively at the C6 position to form an optically pure BcPh 5S,6S-dihydrodiol. However, optically pure BcPh 5R,6S-epoxide was hydrated at both C5 and C6 positions to form a BcPh trans-5,6-dihydrodiol with a (5S,6S):(5R,6R) enantiomer ratio of 32:68.

Animals↗

Reversed-phase high-performance liquid chromatographic separation of phenolic derivatives of benzo[a]pyrene, benz[a]anthracene, and chrysene with monomeric and polymeric C18 columns.

The separation of monohydroxylated derivatives (phenols) of benzo[a]pyrene, benz[a]anthracene, and chrysene was studied by reversed-phase high-performance liquid chromatography using a monomeric Zorbax ODS column and a polymeric Vydac C18 column. The Vydac C18 column resolved the phenols of each hydrocarbon with a wide range of retention times than the Zorbax ODS column. Four K-region phenols of benzo[a]pyrene are either not separated or marginally separated on both monomeric and polymeric columns. Other K-region and non-K-region phenols of all three hydrocarbons can be separated by using the monomeric and polymeric columns in combination.

Animals↗

Stereoselective metabolism of benzo[c]phenanthrene to the procarcinogenic trans-3,4-dihydrodiol.

The enantiomeric compositions of benzo[c]phenanthrene trans-3,4-dihydrodiol, a metabolic precursor of the ultimate carcinogenic bay-region 3,4-dihydrodiol-1,2-epoxides, formed in the metabolism of BcPh by liver microsomes from untreated, phenobarbital (PB)-treated, 3-methylcholanthrene (MC)-treated and polychlorinated biphenyls (PCB; Aroclor 1254)-treated rats of the Sprague-Dawley and Long-Evans strain were each determined by chiral stationary-phase h.p.l.c. and circular dichroism spectral analyses. The percentage of 3R,4R enantiomer in the trans-3,4-dihydrodiol metabolite of BcPh varies, depending on the concentration of liver microsomal enzymes used and the time of incubation: 29-30% (control), 46-54% (PB), 94-100% (MC) and 90-99% (PCB).

Animals↗

Elution order-absolute configuration relationship of K-region dihydrodiol enantiomers of benz[a]anthracene derivatives in chiral stationary phase high-performance liquid chromatography.

The direct resolution of K-region cis- and trans-dihydrodiol enantiomers of 14 unsubstituted and methyl- and bromo-substituted benz[a]anthracene (BA) derivatives was investigated by high-performance liquid chromatography with commercially available columns, packed with gamma-aminopropylsilanized silica to which either (R)-N-(3,5-dinitrobenzoyl)phenylglycine (R-DNBPG) or (S)-N-(3,5-dinitrobenzoyl)leucine (S-DNBL) is either ionically or covalently bonded. BA derivatives used in this study include: BA, 1-methyl-BA, 4-methyl-BA, 7-methyl-BA, 8-methyl-BA, 10-methyl-BA, 11-methyl-BA, 12-methyl-BA, 7,12-dimethyl-BA, 7-bromo-BA, 7-bromo-1-methyl-BA, 7-bromo-11-methyl-BA, 7-bromo-12-methyl-BA, and 3-methylcholanthrene. The enantiomers of BA trans-5,6-dihydrodiol were the only compounds not resolved by any of the four chiral stationary phases (CSPs) tested. The results indicate that conformational preference of the hydroxyl group is one of the most important factor in determining the elution order of dihydrodiol enantiomers. The presence and the location of a substituent and the molecular size and shape of the dihydrodiols can significantly affect the efficiency of enantiomeric resolution. In general, the ionically bonded R-DNBPG provides the best resolution of enantiomeric quasidiequatorial trans-dihydrodiols and the R,R enantiomers are consistently more strongly retained. In contrast, the enantiomeric pairs of quasidiaxial trans-dihydrodiols are generally better resolved by the covalently bonded R-DNBPG, and the S,S enantiomers are more strongly retained. The enantiomers of cis-dihydrodiols having hydroxyl groups that adopt quasiequatorial-quasiaxial and/or quasiaxial-quasiequatorial conformations are more consistently resolved by the ionically bonded S-DNBL and in all cases the S,R enantiomers are more strongly retained. Thus, it is possible to choose a CSP which resolves the K-region dihydrodiol enantiomers with a predictable elution order.

Benz(a)Anthracenes↗

Direct separation of non-K-region mono-ol and diol enantiomers of phenanthrene, benz[a]anthracene, and chrysene by high-performance liquid chromatography with chiral stationary phases.

The direct separation of 26 bay region and non-bay region mono-ol and diol enantiomers of phenanthrene, benz[a]anthracene, and chrysene was compared by high-performance liquid chromatography on commercially available columns, packed with gamma-aminopropylsilanized silica to which either (R)-N-(3,5-dinitrobenzoyl)phenylglycine(R-DNBPG) or (S)-N-(3,5-dinitrobenzoyl)leucine(S-DNBL) was either ionically or covalently bonded. In general, enantiomers of bay region mono-ols and diols are more efficiently resolved than those of non-bay region derivatives. Elution orders of enantiomers on either chiral stationary phase are the same, regardless of whether the chiral stationary phase is ionically or covalently bonded. Except for the enantiomers of 4-hydroxy-4-methyl-1,2,3,4-tetrahydrobenz[a]anthracene, 1,2,3,4-tetrahydrobenz[a]anthracene trans-1,2-diol, and benz[a]anthracene trans-1,2-dihydrodiol, elution orders of resolved enantiomers on R-DNBPG are reversed on S-DNBL. The enantiomers are generally more efficiently resolved on R-DNBPG than on S-DNBL. With the exception of the elution order of the enantiomeric 4-hydroxy-1,2,3,4-tetrahydrochrysene, the results of this study are consistent with the chiral recognition mechanisms proposed by Pirkle and co-workers, who developed the chiral stationary phases used in this study.

Benz(a)Anthracenes↗

Stereoselectivity of rat liver cytochrome P-450 isozymes: direct determination of enantiomeric composition of K-region epoxides formed in the metabolism of benz[a]anthracene and 7,12-dimethylbenz[a]anthracene.

The K-region 5,6-epoxides of benz[a]anthracene (BA) and 7,12-dimethylbenz[a]anthracene (DMBA) were isolated by normal-phase HPLC from metabolites formed by incubation of the respective parent compound with liver microsomes from untreated (control), phenobarbital (PB)-treated, and 3-methylcholanthrene (MC)-treated rats in the presence of an epoxide hydrolase inhibitor, 3,3,3-trichloropropylene 1,2-oxide. The enantiomeric contents of the metabolically formed K-region 5,6-epoxides of BA and DMBA were directly determined by chiral stationary phase HPLC. The K-region 5,6-epoxides formed in the metabolism of BA have (5R,6S): (5S,6R) enantiomer ratios of 25:75 (control), 21:79 (PB), and 4:96 (MC), respectively. In contrast, the (5R,6S):(5S,6R) enantiomeric ratios of the K-region 5,6-epoxides formed in the metabolism of DMBA are 76:24 (control), 80:20 (PB), and 97:3 (MC), respectively. These and earlier results on the stereoselective K-region metabolism studies of 7-methylbenz[a]anthracene and 12-methylbenz[a]anthracene indicate that cytochrome P-450 isozymes exhibit different stereoselectivities in the K-region epoxidations of BA and DMBA and a methyl substituent at the C12 position of BA alters the stereoheterotopic interactions between cytochrome P-450 isozymes and the BA molecule.

9,10-Dimethyl-1,2-benzanthracene↗

Metabolic and stereoselective formations of non-K-region benz(a)anthracene 8,9- and 10,11-epoxides.

The non-K-region benz[a]anthracene (BA) 8,9- and 10,11-epoxides were isolated by normal-phase high-performance liquid chromatography as rat liver microsomal metabolites of BA. The identities of these epoxides were established by ultraviolet and mass spectral analyses and were further validated by the microsomal epoxide hydrolase catalyzed conversion to BA trans-8,9-dihydrodiol and trans-10,11-dihydrodiol, respectively. Circular dichroism spectral analyses of the metabolically formed non-K-region epoxides and dihydrodiols and mass spectral analyses of metabolically formed 18O-labeled non-K-region dihydrodiols and their acid-catalyzed dehydration products indicated that BA (8R,9S)-epoxide and (10S,11R)-epoxide were the predominant enantiomers formed in the metabolism at the 8,9- and 10,11- aromatic double bonds of BA, respectively, by rat liver microsomes. This is the first example demonstrating the direct detection and stereoselective metabolic formation of non-K-region epoxides of a polycyclic aromatic hydrocarbon.

Animals↗