Mapping of a novel SH3 domain protein and two proteins of unknown function to human chromosome 21.

Down syndrome, caused by trisomy of human chromosome 21 (HSA21), is the most common autosomal form of mental retardation. To understand the aetiology of the syndrome we need to identify the genes involved. We have utilised the information generated by the various EST sequencing projects to enrich the transcription map of chromosome 21.

Management of chronic hepatitis C: clinical audit of biopsy based management algorithm.

Objective: To assess the attendance, outcome, compliance with treatment, and response to interferon alfa in patients with chronic hepatitis C who attended during 1995 and were treated according to a biopsy based algorithm.

Design: Retrospective audit of all patients with chronic hepatitis C attending outpatient clinics over one year.

Setting: The liver unit at a London teaching hospital.

Gene transfer using replication-defective human foamy virus vectors.

Replication-defective vectors based on an infectious molecular clone of human foamy virus (HFV) were constructed by deletion and replacement of the accessory genes with expression cassettes for puromycin-resistance and beta-glucouronidase. Cell lines which produced in excess of 10(5) helper virus-free transducing units/ml were generated by trans-complementation of the replication defect using a BHK-21-derived cell line expressing the Bel-1 transactivator. Vectors based on the HFV genome may provide useful alternatives to existing retroviral vectors.

Contiguous arrays of satellites 1, 3, and beta form a 1.5-Mb domain on chromosome 22p.

The centromeric heterochromatin of all the human chromosomes is composed of megabases of tandemly repeated satellite DNA. Some of these sequences have been implicated in centromere formation and/or segregation but the arrangement of most of them on a large scale remains largely uncharacterized because of the difficulties in analyzing repetitive DNA. The alpha satellite is the best studied and is present in large tandem arrays at all centromeres, but satellites 1, 3, and beta have also been detected on a number of chromosomes.

Genital mycoplasma infections.

Since 1937, 13 Mycoplasma species, two Acholeplasma species, and one Ureaplasma species have been isolated from humans. Six of these have the urogenital tract as the primary site of colonisation but others, which have the oropharynx and respiratory tract as the primary site, are found occasionally in the urogenital tract because of orogenital contact. Mycoplasma hominis was the first to be isolated and is most strongly associated with bacterial vaginosis (BV), together with a variety of other bacteria.

Bacterial copper- and zinc-cofactored superoxide dismutase contributes to the pathogenesis of systemic salmonellosis.

Copper/zinc-cofactored superoxide dismutase ([Cu,Zn]-SOD) has been found in the periplasm of many bacterial species but its biological function is unknown. Here we report the cloning and characterization of sodC, encoding [Cu,Zn]-SOD, from Salmonella typhimurium. The predicted protein sequence shows only 58% identity to Escherichia coil SodC, and from this its chromosomal location and its immediate proximity to a phage gene, sodC, in Salmonella is speculated to have been acquired by bacteriophage-mediated horizontal transfer from an unknown donor.

RT-PCR method specific for the detection of transgenic CFTR mRNA in the presence of transgene plasmid DNA and endogenous CFTR mRNA.

A major problem with experiments involving the correction of cystic fibrosis by gene transfer has been reliably detecting the transgenic message and distinguishing this from an endogenous message and from vector DNA. We have exploited the SV40 small T antigen intron present in an expression vector containing the CFTR cDNA (pCFAS) using a primer directed to the spliced sequence to allow specific and precise detection of the transgeneic message.

Oestradiol feedback stimulation of androgen biosynthesis by human theca cells.

This study analysed the effect of oestradiol on basal and LH-stimulated production of androstenedione and progesterone by human theca cells in monolayer culture. Incubations were carried out for either 2 days (seven experiments) or 4 days (four experiments), in the presence or absence of luteinizing hormone (LH), oestradiol (10(-9)-10(-6) M) or inhibin. Medium collected at 48 and 96 h was stored until radioimmunoassay for steroid content.

Spatiotemporal patterns of gamma frequency oscillations tetanically induced in the rat hippocampal slice.

1. We used transverse and longitudinal rat hippocampal slices to study the synchronization of gamma frequency (> 20 Hz) oscillations, across distances of up to 4.5 mm.

Reduced interferon-alpha production by Epstein-Barr virus transformed B-lymphoblastoid cell lines and lectin-stimulated lymphocytes in congenital dyserythropoietic anaemia type I.

The concentrations of interferon-alpha (IFN-alpha) in supernatants from cultures of Epstein-Barr virus (EBV) transformed B-lymphoblastoid cell lines derived from seven patients with congenital dyserythropoietic anaemia (CDA) type I were below the 95% confidence limits for those derived from six healthy subjects. In contrast, the concentrations of IFN-alpha in supernatants from cultures of EBV-transformed lymphoblastoid cell lines derived from four patients with other types of CDA and four patients with hereditary sideroblastic anaemia were normal. Supernatants from cultures of peripheral blood lymphocytes stimulated with phytohaemagglutinin or pokeweed mitogen contained less IFN-alpha when the cells were derived from patients with CDA type I than when derived from healthy subjects.

Measurement of glucose turnover--implications for the study of inborn errors of metabolism.

Aspects of glucose metabolism have been investigated quantitatively employing stable isotopes for 20 year. Use of non-recycling [6,6-2H2] or [U-13C] glucose labels provides a value for total hepatic glucose production (glycogenolysis plus gluconeogenesis). Quantitation of gluconeogenesis with isotopic tracers has itself recently been revisited employing protocols and analytical options that purport to overcome the isotope exchanges (dilution) experienced at the level of oxaloacetate when the rate of incorporation of label into glucose from infused alanine, lactate or pyruvate is monitored.

Antiphospholipid antibodies, infertility and recurrent miscarriage.

Antiphospholipid antibodies are found in 15% of women with recurrent miscarriage. These women have only a 10% live birth rate in subsequent pregnancies in which no pharmacological treatment is given. Pregnancy loss is often attributable to uteroplacental insufficiency subsequent to placental thrombosis.

A longitudinal study of the vaginal flora over a menstrual cycle.

Healthy female volunteers participated in an anonymous study to monitor vaginal flora by taking daily vaginal samples and making a smear for later Gram-staining, as well as recording information on genital symptoms, sexual activity, contraceptive and bathing practices. A modification of Spiegel's criteria was used to categorize the Gram-stained smears, an intermediate category between normal flora and bacterial vaginosis (BV) being recognized. Of the 22 volunteers who completed the study, one was excluded because of pregnancy.

Complementation of null CF mice with a human CFTR YAC transgene.

We have made transgenic mice carrying a 320 kb YAC with the intact human cystic fibrosis transmembrane regulator (CFTR) gene. Mice that only express the human transgene were obtained by breeding with Cambridge null CF mice. One line has approximately two copies of the intact YAC.

E2F1-induced apoptosis requires DNA binding but not transactivation and is inhibited by the retinoblastoma protein through direct interaction.

E2F1 overexpression has been shown to induce apoptosis in cooperation with p53. Using Saos-2 cells, which are null for p53 and lack functional Rb, we have demonstrated that E2F1 overexpression can also induce apoptosis in the absence of p53 and retinoblastoma protein (Rb). E2F1-induced apoptosis can be specifically inhibited by Rb but not mdm2, which is known for its ability to inhibit p53-induced apoptosis.

Modern use of clomiphene citrate in induction of ovulation.

Clomiphene citrate is the treatment of first choice in the management of infertility in normally oestrogenized, anovulatory women (WHO group II). The majority of women with 'pure' anovulatory infertility respond to treatment with clomiphene citrate. The rates of pregnancy and miscarriage are close to those expected in a normal fertile population.

Evidence for a primary abnormality of thecal cell steroidogenesis in the polycystic ovary syndrome.

Objective: Polycystic ovary syndrome (PCOS) is a common endocrinopathy of unknown aetiology. The aims of this study were to identify whether ovarian thecal cell steroidogenesis is abnormally regulated in PCOS by measuring steroid responses to a single dose of hCG before, and during, suppression of endogenous LH levels by GnRH analogue (GnRHa).

Design: Serum levels of LH, FSH, 17 alpha hydroxyprogesterone (17OHP), androstenedione, testosterone and dehydroepiandrosterone sulphate were measured before, and 48 hours after, a single intramuscular injection of 10,000 IU hCG.

Expression of serotonin transporter mRNA in rat brain: presence in neuronal and non-neuronal cells and effect of paroxetine.

The expression of serotonin transporter mRNA in rat brain was-examined by in situ hybridisation. Hybridisation was observed in cells of the known serotonergic nuclei and no other neuronal populations. It was also associated with ependymal cells of the aqueduct which may indicate a specialisation of this part of the ventricular system in anatomical and neurophysiological terms.

Host genetic factors influencing the outcome of hepatitis.

A strong genetic component determining the outcome of hepatitis B virus (HBV) infection has been established through twin studies. The immunopathogenesis of HBV infection is well described and it has therefore been possible to predict some gene loci, exhibiting polymorphism, may influence the outcome of HBV infection. As expected, the immune response genes in the major histocompatibility complex (MHC) on the short arm of chromosome 6 have provided confirmed susceptibility genes.

Relationship between hydrogen peroxide-producing strains of lactobacilli and vaginosis-associated bacterial species in pregnant women.

This study was conducted to determine the relationship between lactobacilli and bacterial species associated with bacterial vaginosis in pregnancy and the prevalence of H2O2-producing and non-producing strains of lactobacilli in pregnant women whose vaginal flora had already been analysed. Information was available for 174 pregnant women whose vaginal flora had been evaluated previously by examining gram-stained vaginal smears: 50 had grade III flora (bacterial vaginosis). 50 grade II flora, 41 flora graded as abnormal which then reverted to grade I (revertants) and 33 normal flora (controls).