The oxidation of benzo[a]pyrene (B[a]P) was examined using reconstituted systems prepared with recombinant human cytochrome P450 (P450) enzymes 1A1, 1A2, 2C8, 2C10, 2E1, and 3A4 and with microsomes prepared from Saccharomyces cerevisiae expressing recombinant human P450s 2C8, 2C9, and 2C18. Products measured by HPLC included the 3- and 9-phenols, the 4,5-, 7,8-, and 9,10-dihydrodiols (detected in the presence of epoxide hydrolase), and products in the polar fraction eluting immediately after the void volume. The most active enzyme in all reactions was P450 1A1. P450 3A4 and P450 1A2 formed appreciable amounts of several of the products, including the 3-phenol. P450 2C enzymes and P450 2E1 formed relatively low amounts of all B[a]P products. Consideration of these patterns along with knowledge of levels of expression of the P450s in human tissues and previous results with microsomes leads to the conclusion that P450 1A1 should dominate the oxidation of B[a]P in tissues where it is present and inducible. In human liver the level of P450 1A1 is low and P450 3A4, P450 2C subfamily enzymes, and P450 1A2 probably all contribute. Of the human P450s considered here, P450 1A2 was the most active hepatic enzyme forming the 7,8-dihydrodiol. 7,8-Benzoflavone stimulated the oxidation of B[a]P by P450 3A4 and inhibited the oxidations catalyzed by P450 1A2. The extent of inhibition of P450 1A1 was less (than with P450 1A2), probably due to the rapid oxidation of 7,8-benzoflavone by P450 1A1. The major 7,8-benzoflavone product appears to be the 5,6-oxide.

Benzo(a)pyrene↗

PubMedDate not supplied

The sulfoxidation of the hexachlorobutadiene metabolite N-acetyl-S-(1,2,3,4,4-pentachlorobutadienyl)-L-cysteine is catalyzed by human cytochrome P450 3A enzymes.

The sulfoxidation of the mercapturic acid N-acetyl-S-(1,2,3,4,4-pentachlorobuta-1,3-dienyl)-L-cysteine (N-Ac-PCBC), a urinary metabolite of the renal toxin hexachlorobutadiene (HCBD), was studied in human liver microsomes and with purified cDNA expressed human liver cytochrome P450 (P450) enzymes. N-Acetyl-S-(1,2,3,4,4-pentachlorobuta-1,3-dienyl)-L-cysteine sulfoxide (N-Ac-PCBC SO) is a major urinary metabolite of HCBD in male rats; only liver microsomes from male rats catalyze the sulfoxidation of N-Ac-PCBC. Our results presented here show that human liver microsomes from both male and female donors are capable of oxidizing N-Ac-PCBC to the corresponding sulfoxide diastereomers. The correlation of N-Ac-PCBC sulfoxidation with the rates of oxidation of P450 enzyme specific substrates suggests that only P450 3A enzymes oxidize N-Ac-PCBC. Moreover, only gestodene and troleandomycin, two selective inhibitors for P450 of the 3A family, significantly reduced the rates of N-Ac-PCBC sulfoxidation. No reduction in sulfoxidation rates was observed with inhibitors for other P450 enzymes, i.e., diethyldithiocarbamate, 4-methylpyrazole, 7,8-benzoflavone, or sulfaphenazole. Incubations of N-Ac-PCBC with purified and reconstituted recombinant P450s 1A2, 2E1, 3A4, and 3A5 resulted in sulfoxide formation only with P450s 3A4 and 3A5. In summary, these results indicate that P450 from the 3A family may sulfoxidize N-Ac-PCBC. Since these P450 enzymes account for a major fraction of the P450 in human liver and are also present in human kidney, the sulfoxidation reaction may also be expected to occur in humans exposed to HCBD.

Acetylcysteine↗

PubMedDate not supplied

Active site topology of human cytochrome P450 2E1.

Cytochrome P450 2E1 (CYP2E1), an enzyme that is induced by ethanol, plays an important role in the metabolism of various toxic and carcinogenic substances, including dialkylnitrosamines and small halocarbons. Little information is available on the active site of the enzyme. We report here the formation of aryl-iron complexes (Fe-Ar) in the reactions of human CYP2E1 with phenyldiazene, (2-naphthyl)hydrazine, and p-biphenylhydrazine. Migration of the aryl group from the iron to the porphyrin nitrogens within the intact active site produces the four possible N-arylprotoporphyrin IX regioisomers in the following ratios (NB:NA:NC:ND, where the subscript indicates the pyrrole ring): phenyl, 03:34:02:61; 2-naphthyl, 02:18:03:77; p-biphenyl, 02:52:04:42. The results indicate that the active site of human CYP2E1 is sterically unhindered directly above the iron for a distance of 10 A. It appears, furthermore, that the active site cavity is relatively open above pyrrole rings A and D but is closed above pyrrole rings B and C, a topology similar to that deduced for the rat enzyme.

Animals↗

PubMedDate not supplied

Relation between commuting, leisure time physical activity and serum lipids in a Chinese urban population.

PRIMARY OBJECTIVE: To analyse the association between both commuting and leisure time physical activity and serum lipids in a Chinese population. POPULATION: A total of 1786 males and 1922 females aged 20-49 years. METHODS: A cross-sectional survey was carried out in 1989 in urban area of the city of Tianjin, China. Commuting and leisure time physical activity, total cholesterol, high density lipoprotein cholesterol, low density lipoprotein cholesterol and triglycerides were determined. RESULTS: Daily walking or cycling to and from work was inversely associated with serum total cholesterol, low-density lipoprotein cholesterol and triglyceride concentrations among men and positively associated with high-density lipoprotein cholesterol concentrations among women as compared to travelling to and from work by bus. CONCLUSIONS: The present study showed a significant association between serum lipids and commuting physical activity, but no relation with leisure time sports.

Adult↗

PubMedDate not supplied

Synthesis of 2',3'-dideoxy-2',3'-didehydro nucleosides via a serendipitous route.

This paper describes a "green" synthesis of 2',3'-unsaturated 2',3'-dideoxynucleosides via an electrochemical reaction. Using this approach d4T, d4U, ddA and ddI can be synthesized in high yields.

Dideoxynucleosides↗

PubMedDate not supplied

Delayed radiation encephalopathy after radiotherapy for nasopharyngeal cancer: a CT study of 45 cases.

The CT features of 45 cases of delayed radiation encephalopathy (including radiation necrosis) following radiotherapy for nasopharyngeal carcinoma are reported. The brain lesions were uni- or bilateral and involved mainly the white matter and subsequently the gray matter of the lower portion of the brain included within the portals of irradiation and its vicinity. The lesions were edematous and hypodense on CT and showed postcontrast enhancement in 50% of the cases. Within the period of follow-up (1-5 years), the lesions showed remissions and exacerbations and in some cases stabilized. In addition, there was progressive cerebral atrophy, manifesting itself mainly as dilatation of the temporal horns, the neighboring cisterns, and sylvian fissures. In some cases that were followed for a long time, the cerebral lesions showed either foci of calcification or encephalomalacia and/or porencephaly.

Adrenal Cortex Hormones↗

PubMedDate not supplied

Efficient gene transfer and expression in islets by an adenoviral vector that lacks all viral genes.

Although adenoviral vector-mediated gene transfer has significant potential for gene therapy, host immune responses to virally expressed proteins and small insert capacity may limit its clinical application. In order to overcome these disadvantages, a new adenoviral vector that lacks all viral genes has been developed. Using the green fluorescent (GFP) gene as a reporter gene, we investigated the efficiency of gene transfer by this all-viral-genes-deleted and minimal cis-element remaining adenoviral vector (miniAd-GFP) in islets in vitro and ex vivo, and compared it with the E1-deleted adenoviral vector (E1-GFP). One day after in vitro infection, GFP was expressed in both miniAd-GFP- and E1-GFP-infected islets. The percentage of GFP-positive single cells was not significantly different between miniAd-GFP-infected islets and E1-GFP-infected islets. When these islets were transplanted into syngeneic diabetic mice, both miniAd-GFP- and E1-GFP-infected islet grafts reversed diabetes, and normal blood glucose levels were maintained for over 20 weeks posttransplantation. Mild lymphocyte infiltration was found in all E1-GFP-infected islet grafts at all time points. However, this was not seen in most miniAd-GFP-infected islet grafts. Our results indicate that gene transfer by an adenoviral vector that lacks all viral genes is as efficient as E1-deleted adenoviral vector-mediated gene transfer in islets. Furthermore, this adenoviral vector might be less immunogeneic than the E1-deleted adenoviral vector.

Adenoviridae↗

PubMedDate not supplied

Immunohistochemical and DNA sequencing analysis on human mismatch repair gene MLH1 in cervical squamous cell carcinoma with LOH of this gene.

BACKGROUND: The human MLH1 gene (hMLH1) is one of the DNA mismatch repair genes. Defects in these genes are believed to be the underlying cause of microsatellite instability (MSI). MSI has been demonstrated in many human cancers such as colon cancer and some female-specific tumors. The hMLH1 gene can be inactivated by genetic mutation or by hypermethylation of its promoter region, which often causes cessation of hMLH1 protein production. We were prompted by our previous finding of 43% of cervical cancers and their precursors with a loss of heterozygosity (LOH) of the hMLH1 gene to investigate whether it is inactivated during the carcinogenesis of cervical cancer. MATERIALS AND METHODS: hMLH1 protein production was assessed by immunohistochemistry, and nucleotide sequence analysis were performed on all exons of the hMLH1 gene. RESULTS: In 11 cases of invasive cervical cancer, among which 7 had LOH of the hMLH1 gene, immunohistochemistry provided no evidence for cessation of hMLH1 protein production. In addition, no mutations were found in any of the 19 hMLH1 gene exons. CONCLUSIONS: The activity of hMLH1 gene might not be disturbed in the development of cervical cancer although a proportion of the cases had lost one of its hMLH1 gene copies.

Adaptor Proteins, Signal Transducing↗

PubMedDate not supplied

[Cervical carcinoma progression-associated genetic alterations on chromosome 6].

To identify the loci associated with progression of cervical carcinoma, chromosome 6 regions were tested for loss of heterozygosity. Detailed analysis with 28 microsatellite markers revealed a high frequency of allelic deletions for several loci of the short (6p25, 6p22, 6p21.3) and long (6q14, 6q16-21, 6q23-24, 6q25, 6q27) arms of chromosome 6. Examination of 37 microdissected carcinoma and 22 cervical dysplasia specimens revealed allelic deletions from the HLA class I-III genes (6p22-21.3) and subtelomeric locus 6p25 were found in more than 40% dysplasia specimens. With multiple microdissection of cryosections, genetic heterogeneity of squamous cervical carcinoma was analyzed, and clonal and subclonal allelic deletions from chromosome 6 were identified. Half of the tumors had clonal allelic deletion of D6S273 (6p21.3), which is in a Ly6G6D (MEGT1) intron in the HLA class III gene locus. The frequency of allelic deletions from the chromosome 6 long arm was no more than 20% in dysplasias. Allelic deletions from two loci, 6q14 and 6q16-21, were for the first time associated with invasion and metastasis in cervical carcinoma.

Chromosomes, Human, Pair 6↗

PubMedDate not supplied

The composition of Pneumocystis carinii phospholipids.

Explore the source record for details and available documents.

Animals↗

PubMedDate not supplied

Binding characteristics of glucocorticoid receptor in synaptic plasma membrane from rat brain.

The study was carried out to identify the specific glucocorticoid membrane binding site (GCMB) on synaptic plasma membrane (SPM), and to investigate the ligand binding properties and physico-chemical characteristics of GCMB. The radioligand binding assay showed that tritiated corticosterone ([3H]B) could bind to GCMB specifically with positive cooperativity. For SPM, the binding capacity (R(zero)) was 46.67 +/- 4.62 fmol/mg protein; the dissociation constant (Kd), 97.48 +/- 18.85 nmol/L, and the Hill coefficient, 1.633 +/- 0.082. Various steroids bound with GCMB in the following rank order: RU26752, aldosterone > corticosterone, cortisol > RU26988, progesterone, R5020, triamcinolone acetonide > testosterone, estradiol > RU38486 > dexamethasone. GCMB, which could be solubilized by detergent Triton X-100, showed thermal stability, although it was susceptible to hydrolysis by trypsin and to the reducing agent, mercaptoethanol. The addition of divalent cations (Ca2+, Mg2+) was beneficial to the ligand binding. The results strongly indicate that GCMB, fulfilling the basic criteria for a receptor, constitutes a novel type of glucocorticoid receptor on neuronal membrane which is significantly different from the cytosolic glucocorticoid receptors.

Animals↗