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Biomedical subjects

E Jeffery

Publications and source records attributed to E Jeffery.

12 recordsLinked to original sources
PubMed1999

Monitoring gene expression profile changes in ovarian carcinomas using cDNA microarray.

The development of cancer is the result of a series of molecular changes occurring in the cell. These events lead to changes in the expression level of numerous genes that result in different phenotypic characteristics of tumors. In this report we describe the assembly and utilization of a 5766 member cDNA microarray to study the differences in gene expression between normal and neoplastic human ovarian tissues. Several genes that may have biological relevance in the process of ovarian carcinogenesis have been identified through this approach. Analyzing the results of microarray hybridizations may provides new leads for tumor diagnosis and intervention.

Cloning, Molecular↗
PubMed1998

Alcohol-mediated increases in acetaminophen hepatotoxicity: role of CYP2E and CYP3A.

This commentary focuses on the roles of CYP3A and CYP2E in alcohol-mediated increases in acetaminophen hepatotoxicity. CYP2E has been considered to be the main form of P450 responsible for such toxicity in animals and humans. However, CYP3A, which is also induced by alcohol, has been shown to have a greater affinity for acetaminophen than CYP2E. Previous experiments implicating CYP2E in alcohol-mediated increases in acetaminophen hepatotoxicity have used inhibitors of this form of P450 that are now proving to be non-specific. Triacetyloleandomycin (TAO) is a potent inhibitor of CYP3A that maintains specificity in vitro over a large concentration range. In rats treated with ethanol or the combination of ethanol and isopentanol, the major higher chain alcohol in alcoholic beverages, TAO protects animals from increases in acetaminophen hepatotoxicity, suggesting a major role of CYP3A. CYP2E may not have a major role due to the rapid loss of induced levels in the absence of continued exposure to ethanol. Knockout mice, which are being used to define the role of particular proteins in biological responses, have been developed for CYP2E1 and CYP1A2 but not CYP3A. Cyp2e1(-/-) and Cyp1a2(-/-) mice are more resistant to acetaminophen hepatotoxicity than wild-type strains, even though the amounts of the other forms of P450s are unaltered in the liver. These findings suggest that the relative amounts of P450s and not just kinetic characteristics determine their role in acetaminophen hepatotoxicity. The clinical implications of the findings that CYP3A can have a major role in acetaminophen-mediated hepatotoxicity are discussed.

Acetaminophen↗
PubMed1996

Studies on the toxic effects of crambe meal and two of its constituents, 1-cyano-2-hydroxy-3-butene (CHB) and epi-progoitrin, in broiler chick diets.

1. Studies were undertaken to determine a safe inclusion rate for crambe (Crambe abyssinica) meal in broiler chick diets, and to determine the mechanism for adverse effects by investigating its constituents; 1-cyano-2-hydroxy-3-butene (CHB) and 3-butenyl glucosinolate (epi-progoitrin, E-PG). 2. Crambe meals were prepared to differ in E-PG (19, 36 and 40 g/kg) and CHB contents (0.1, 0.7 and 1.9 g/kg), and with either active or inactive thioglucosidase. 3. Meals were fed to 7-d-old broiler chicks at 50 or 100 g/kg of the diet for 12 or 13 d. In separate studies, isolated E-PG or CHB were mixed into the diet or administered by gavage to 7-d-old broiler chicks in amounts equivalent to 50 or 100 g/kg crambe meal diets for 10 and 12 d, respectively. 4. Weight gain decreased (P < 0.05) in chicks fed on the high glucosinolate crambe diets or isolated E-PG. Food consumption decreased (P < 0.05) in chicks fed on the diet containing the high E-PG meal with active enzyme. 5. Mild liver lesions and increased serum aspartate aminotransferase were found in chicks fed on the diet containing the high glucosinolate meal with active enzyme. Other organs, including thyroids, were normal. 6. Commercially-processed crambe meal appeared safe at an inclusion rate of 50 or 100 g/kg diet, but could not be recommended at this point for long term feeding.

Alkenes↗
PubMed1993

Close proximity of the genes for leukemia inhibitory factor and oncostatin M.

Leukemia inhibitory factor (LIF) and oncostatin M (OSM) are structurally related cytokines that share a common receptor complex. The genes for LIF and OSM have previously been reported to be on human chromosome 22. We report here the isolation of a single 73 kbp human genomic yeast artificial chromosome clone that contains the LIF and OSM genes. The two genes lie within 19 kbp of each other and are transcribed in opposite directions.

Base Sequence↗
PubMed1991

Enhanced hematopoietic activity of a human granulocyte/macrophage colony-stimulating factor-interleukin 3 fusion protein.

Granulocyte/macrophage colony-stimulating factor-interleukin 3 (GM-CSF-IL-3) fusion proteins were generated by construction of a plasmid in which the coding regions of human GM-CSF and IL-3 cDNAs were connected by a synthetic linker sequence followed by subsequent expression in yeast. Both GM-CSF-IL-3 and IL-3-GM-CSF fusion proteins were purified to homogeneity and shown to bind to cell-surface receptors through either their GM-CSF or IL-3 domains. The fusion proteins exhibited enhanced receptor affinity, proliferative activity, and hematopoietic colony-stimulating activity compared with either IL-3 and/or GM-CSF alone. This suggests that GM-CSF-IL-3 fusion proteins may hold future promise as therapeutic agents.

Base Sequence↗
PubMed1991

Experimental and theoretical studies of the three-dimensional structure of human interleukin-4.

The structure of human interleukin 4 (IL-4) was predicted utilizing a series of experimental and theoretical techniques. Circular Dichroism (CD) spectroscopy indicated that IL-4 belonged to the all alpha-helix class of protein structures. Secondary structure prediction, site-directed mutagenesis, and CD spectroscopy suggested a predominantly alpha-helical structure, consistent with a four-helix bundle structural motif. A human/mouse IL-4 chimera was constructed to qualitatively evaluate alternative secondary structure predictions. The four predicted helices were assembled into tertiary structures using established algorithms. The mapping of three disulfide bridges in IL-4 provided additional constraints on possible tertiary structures. Using accessible surface contact area as a criterion, the most suitable structures were right handed all antiparallel four-helix bundles with two overhand loop connections. Successful loop closure and incorporation of the three disulfide constraints were possible while maintaining the expected shape, solvent accessibility, and steric interactions between loops and helices. Lastly, energy minimization was used to regularize the chain.

Amino Acid Sequence↗
PubMed1990

Effect of fibronectin on the Crithidia luciliae test for anti-double-stranded DNA antibodies.

The various tests for anti-double-stranded DNA antibodies do not always agree. Plasma fibronectin specifically binds DNA, is a component of immune complexes, and shows variations in concentration with disease activity in systemic lupus erythematosus. It may therefore interfere with the detection of DNA autoantibodies. This possibility was examined in a series of studies using the Crithidia luciliae test. Studies were based on serum samples received during one year (250 samples). Serum samples from 50 patients which were positive or weakly positive in the C luciliae test were used. In blocking experiments fibronectin was added either to the wells or to the serum. In a second series of experiments fibronectin was depleted by affinity chromatography from six serum samples with weak anti-DNA staining. Preincubation of wells with fibronectin or addition of fibronectin to serum invariably blocked the interaction of anti-DNA antibodies with the C luciliae kinetoplast. When fibronectin was removed from serum the intensity of staining was increased. These results indicate that fibronectin influences the detection of anti-double-stranded DNA antibodies using C luciliae and may explain the disparity between the results of different tests for DNA antibodies. Furthermore, the unmasking of positive reactivity when fibronectin is removed from serum has implications for the diagnosis and treatment of systemic lupus erythematosus.

Animals↗
PubMed1987

Lack of hepatic microsomal metabolism of deoxynivalenol and its metabolite, DOM-1.

Rat hepatic microsomal preparations were used to study the metabolism of deoxynivalenol (DON) and its metabolite 3 alpha,7 alpha,15-trihydroxytrichothec-9,12-dien-8-one (DOM-1). The N-demethylation of ethylmorphine was monitored to assess the viability of the mixed-function oxidase. DON was incubated with microsomes and an NADPH-generating system. Samples were removed from the incubation system and analysed for DON using an HPLC equipped with a UV detector. After incubation for 30 min, there was no evidence of disappearance of DON or of the presence of new metabolites; neither was microsomal NADPH oxidation altered by the addition of DON. Rat and pig hepatic microsomal preparations were used to assess DON glucuronidation, using p-nitrophenol disappearance to check the viability of the microsomal glucuronidating system. When DON was incubated with microsomes and 14C-labelled uridine 5'-diphosphoglucuronic acid, no radioactivity was detected in the TLC zone where the glucuronide was expected. Three rats and one pig were dosed orally with 2 mg DON/kg and samples of their urine and faeces were extracted and incubated with beta-glucuronidase or with buffer only. No differences in DON or DOM-1 concentrations were detected between samples incubated with or without beta-glucuronidase. These results suggest that DON was neither bioactivated to a more toxic product nor oxidized to a less toxic compound by the rat hepatic mixed-function oxidase system. Likewise, DOM-1 was not reactivated or metabolized by this system. Neither DON nor DOM-1 glucuronides were formed either in in vitro liver systems or in vivo.

Animals↗
PubMedDate not supplied

Comparison of hepatic microsomal drug-metabolizing systems from rats fed crude and purified diets.

Hepatic microsomes from rats fed a crude or a purified diet were compared by measureing their contents of protein, cytochrome P-450, and cytochrome b5, their rates of activity of NADPH- and NADH-cytochrome c reductases, NADPH-cytochrome P-450 reductase, NADPH oxidase, lipid peroxidase, ethylmorphine N-demethylase, aniline hydroxylase, benzpyrene hydroxylase, and their substrate-binding spectra (ethylmorphine, hexobarbital, aniline, and ethyl isoyanide). With the exception of lipid peroxidase activity, which was much higher in microsomes from animals fed the crude diet, little or no consistent diet-related differences in these measurements were observed over a 4-week experimental period, nor were results significantly less variable with one or the other diet. No consistent significant differences were observed with two strains of rats. The lower lipid peroxidase activity seen with the purified diet appeared to be due to the high vitamin E intake when that diet was employed; rats fed the crude diet and an oral supplement of alpha-tocopherol yielded microsomes with low lipid peroxidase activities similar to those seen in microsomes from rats fed the purified diet. A gradual temporal increase in benzpyrene hydroxylase activity was observed with both diets. This was interpreted to be due to environment inducing agents other than those present in the diet.

Analysis of Variance↗