Structural Requirements for the Direct and Cytochrome P450-dependent Reaction of Cyclic α,β-Unsaturated Carbonyl Compounds with Glutathione: a Study with Coumarin and Related Compounds

Abstract— The interaction of glutathione (GSH) with coumarin, or one of a series of compounds related to coumarin, was assessed in the absence and presence of liver microsomes (direct reaction and indirect reaction, respectively) to determine the structural requirements for direct and mono-oxygenase-mediated reaction of cyclic α,β-unsaturated carbonyls with GSH. Acrolein was used as a positive control for the direct reaction, and produced complete or nearly complete depletion of GSH under all assay conditions. 5,6-Dihydro-2H-pyran-2-one and 2-cyclohexen-1-one also produced substantial depletion of GSH in the direct reaction, which was not increased by the addition of liver microsomes. Coumarin, 2H-pyran-2-one and precocene I (a substituted pyran lacking the 2-one structure) were not substrates for the direct reaction but did cause depletion of GSH when incubated in the presence of rat or human liver microsomes. These depletions were dependent on a functioning mono-oxygenase system as judged by the effects of omission of cofactors, addition of competitive or inactivating inhibitors of cytochrome P450, and induction. Dihydrocoumarin, ?-valerolactone, cyclohexanone and 4H-pyran-4-one were not substrates for either the direct or indirect reaction. These findings are rationalized on the basis of a direct nucleophilic attack of GSH on the α,β-centre of the α,β-unsaturated carbonyl compounds, which is hindered by benzenoid resonance in coumarin and 2H-pyran-2-one, for which enzyme-mediated reaction with GSH, probably via a 3,4-epoxide, is the favoured mechanism.     

A survey of forensic psychologists’ work with sex offenders in England and Wales

This paper describes the results of a survey of the extent and variety of work with sex offenders by psychologists working in different forensic settings in England and Wales. A postal questionnaire was sent out to all 199 psychologists in the special hospitals, regional forensic psychiatry services, prisons and youth treatment service requesting details about their work with sex offenders during the year June 1991 to July 1992. The results are described under the headings of assessment and treatment; supervision, consultation, teaching and training; research and evaluation. The discussion highlights some of the patterns and trends of the work of forensic psychologists with sex offenders, together with the range of psychological interventions. There are also patterns within services. Results of the survey highlight the links psychologists have with other disciplines in working with sex offenders and raise questions about the most appropriate model of psychology practice, given the limited resources.     

Evaluation of an Australian intervention to encourage breast feeding in primiparous women

Breast feeding has nutritional, immunologic and antiallergicadvantages for the infant. Although it has been widely recommendedthat infants be exclusively breast fed until 4–6 monthsof age, only about half of all Australian babies currently receiveextended breast feeding. The present study evaluated an intensiveprogramme designed to increase the proportion ofprimi-parousmothers who breast fed for 4 months or longer. Women who registeredwith the hospital at least 20 weeks before delivery and whointended to breastfeed were eligible for the study. Two hundredand thirty-five women were allocated to receive either usualcare or an intensive breast feeding programme when they registeredwith the hospital. The intensive programme consisted of writtenmaterials, and group and individual sessions with a lactationcounsellor. It also included a visit from a breastfeeding consultantwhile in hospital after the birth and contact on return home.Women were followed up 6 weeks and 4 months after delivery.There were no significant differences in breastfeeding ratesbetween the control and intervention groups at either follow-uppoint. Breast feeding until 4 months was more likely among womenwhose baby did not receive a bottle feed while still in hospitaland who did not smoke, use the combined oral contraceptive pillor introduce solid food before 4 months. Those mothers who enjoyedand felt satisfied with breastfeeding were more likely to continueto 4 months. It seems likely that programmes designed to increasebreastfeeding will need to address underlying factors such ashospital policy rather than simply providing more health education.     

A sensitive micro-immunoassay using beta-galactosidase/anti-beta-galactosidase complexes

This paper describes a new sensitive microELISA based on enzyme/anti-enzyme complexes following an unlabelled antibody bridging step. beta-Galactosidase/anti-beta-galactosidase complexes were made using a monoclonal antibody raised against bacterial (E. coli) beta-galactosidase and enzyme activity was quantified with a fluorogenic substrate. Because of its high sensitivity the assay is particularly suitable for the detection of limited amounts of antigen. One application illustrated is the analysis of Class I and Class II histocompatibility antigens on peripheral blood lymphocytes using 5000 cells/well in 60-well Terasaki or 96-well microtitre plates.     

HLA Polymorphism in Israel

Phenotype and gene frequencies of antigens at the A and B HLA loci were determined in a sample of 89 random Libyan Jews now settled in Israel. Most antigens at the A locus are within the range found in European populations while, at the B locus, the frequencies differ from Caucasoid populations: B7 is usually rare (1%), Bw35 is frequent (14%) and Bw40 very common with more than 10%. Thirteen percent of Libyan women whose sera were screened for the presence of HLA antibodies gave positive results. Some of them are elderly women, about 15 years or more beyond their last birth.     

Disease associations and altered immune function in CD45 138G variant carriers

The CD45 antigen is a haemopoietic cell specific tyrosine phosphatase essential for antigen receptor mediated signalling in lymphocytes. Expression of different patterns of alternatively spliced CD45 isoforms is associated with distinct functions. We recently identified a polymorphism in exon 6 (A138G) of the gene encoding CD45 (PTPRC) that results in altered CD45 splicing. The 138G allele is present at a high frequency among Japanese (23.7%), with 5.1% individuals homozygous for the G allele. In this study we show that the A138G polymorphism is the cause of altered CD45 isoform expression, promoting splicing towards low molecular weight CD45 isoforms. We further report that the frequency of A138G heterozygotes is significantly reduced in number in cohorts of patients with autoimmune Graves' disease or hepatitis B infection, whereas G138G homozygotes are absent from a cohort of Hashimoto's thyroiditis patients. We also show that 138G individuals exhibit altered cytokine production in vitro and an increased proportion of memory T cells. These data suggest that the 138G variant allele strongly influences these diseases by modulation of immune mechanisms and may have achieved its high frequency as a result of a natural selection probably related to pathogen resistance.