Effects of two antiandrogen treatments on hirsutism and insulin sensitivity in women with polycystic ovary syndrome

Thirty-two women with polycystic ovary syndrome (PCOS) wereallocated to two antiandrogen treatment regimens; 28 women completedthe trial. Twenty women were treated with ethinyloestradioland cyproterone acetate (EO-CA) cyclically for 6 months andeight women were treated with the gonadotrophin releasing hormone(GnRH) analogue, goserelin for 6 months. Effects on hirsutism,insulin sensitivity (estimated by glucose clamp technique),blood lipids and hormones were measured. Women treated withEO-CA showed a reduction in hirsutism (P <0.05), and decreasedserum androgen concentrations (P <0.001) as well as reducedinsulin sensitivity (P <0.05). In women treated with goserelin,serum androgen concentrations also decreased (P <0.001),but there was no significant reduction of hirsutism. This group,however, showed an improved insulin sensitivity (P <0.05)despite an unchanged body mass index. Bone mineral density wasunaltered in both treatment groups. The reduction in androgenconcentrations caused by EO-CA was not paralleled by increasedinsulin sensitivity, most probably due to the effect of ethinyloestradiolper se. In contrast, the reduction in androgen concentrationsby goserelin was accompanied by an improved insulin sensitivity.     

No germline TP53 mutations detected in familial and bilateral testicular cancer

Mutations in the TP53 gene are considered to be among the most common genetic alterations in human cancers. Both somatic and germline mutations have been found. Using potymerase chain reaction (PCR), constant denaturant gel electrophoresis (CDGE), and denaturing gradient gel electrophoresis (DGGE), we have examined 32 patients with bilateral and familial germ cell tumors (GCT) and two patients with sporadic GCT for germline mutations within the conserved regions of the gene. In addition, 15 tumors were screened for somatic mutations and analyzed for loss of heterozygocity (LOH) at the TP53 locus. Twelve tumors were analyzed for expression of TP53 via immunohistochemistry. Neither germline nor somatic TP53 mutations were deteeted. LOH was observed in one of five informative cases. No tumors showed increased expression of TP53 protein. These results indicate that alterations in the TP53 gene are not important for the predisposition to and development of GCT. © 1993 Wiley-Liss, Inc.     

Linkage and association of adrenergic and dopamine receptor genes in the distal portion of the long arm of chromosome 5 with systolic blood pressure variation

Elevated blood pressure is an important risk factor for renal-, cerebro- and cardiovascular diseases. We used an efficient discordant sib-pair ascertainment scheme to investigate the impact of the distal end of the long arm of human chromosome 5 (chromosomal region 5q31.1-qter) containing genes for the alpha1B and beta2 adrenergic receptors and the dopamine receptor type 1A on variation of systolic blood pressure in young Caucasians. We measured eight highly polymorphic markers spanning this positional candidate gene-rich region in 427 individuals from 55 three-generation pedigrees containing 69 discordant sibling pairs, and calculated multipoint identity by descent (MIBD) probabilities. The results of genetic linkage and association tests indicate that the region between markers D5S2093 and D5S462 is significantly linked to one or more polymorphic genes influencing interindividual variation in systolic blood pressure levels. Since the alpha1B adrenergic receptor and dopamine receptor type 1A genes are located close to these markers, these data suggest that genetic variation in one or both of these G protein-coupled receptors, which participate in the control of vascular tone, plays an important role in influencing interindividual variation in systolic blood pressure levels.      

Cytomegalovirus (CMV) antigenemia assay is more sensitive than shell vial cultures for rapid detection of CMV in polymorphonuclear blood leukocytes.

We compared the cytomegalovirus (CMV) antigenemia assay with shell vial cultures of polymorphonuclear leukocyte (PMNL)-enriched blood fractions for rapid diagnosis of CMV viremia. PMNL fractions of 280 blood specimens from 171 patients (170 solid-organ transplant recipients and 1 patient undergoing pretransplant evaluation) were inoculated in shell vial and conventional CMV cultures. A commercially available kit (CMV-vue kit; INCSTAR Corp.) was used for the CMV antigenemia assay, in which PMNL preparations were stained with monoclonal antibodies directed against the CMV protein pp65. Mixed-leukocyte blood fractions from the same blood specimens were inoculated in parallel shell vial and conventional cultures. CMV viremia (defined by the isolation of CMV in conventional cultures) was detected in 32 (13%) of 245 PMNL fractions included in the final analysis. Twenty-eight (87.5%) were also positive in the CMV antigenemia assay, whereas 22 (69%) were positive in shell vial cultures. Ten (4%) additional PMNL fractions positive only in the CMV antigenemia assay were from eight patients with active CMV infections (six patients), who had previous or subsequent episodes of CMV viremia (seven patients), or in whom CMV was isolated in cultures of simultaneously obtained mixed-leukocyte fractions (three patients). Overall, the CMV antigenemia assay was significantly more sensitive than shell vial cultures for detection of CMV in the PMNL fraction of blood leukocytes (P < 0.01, McNemar's test), and we recommend it as the method of choice for rapid diagnosis of CMV viremia.     

Density profile of group B streptococci, type III, and its possible relation to enhanced virulence.

The buoyant densities of virulent and colonizing group B streptococci, type III, were determined by centrifugation of bacteria on a linear, hypotonic density gradient. A total of 28 strains were investigated. Eleven strains were obtained from blood cultures of babies with early-onset disease, and eight strains were isolated from the cerebrospinal fluid of babies with late-onset septicemia and meningitis. Nine colonizing strains were genital isolates from pregnant women subsequently giving birth to healthy children. In each strain the buoyant density was determined before and after neuraminidase treatment. All strains showed an increase in the buoyant density after enzymatic removal of sialic acid, and the density differences before and after desialylation were calculated. The mean values of these differences for blood, cerebrospinal fluid, and colonizing isolates were 23.4, 25.3, and 10.6 mg/ml, respectively. The mean value for the colonizing strains differed significantly from the mean value for each group of virulent strains. All colonizing strains banded singly in the gradient, whereas five of the virulent strains divided into two density populations. Extracts of the low-density cells produced markedly more dense immunoprecipitates with type antiserum than did extracts of the high-density bacteria. One double-banding strain was positive for R protein. After separation of the two density populations, this antigen was detected only in the low-density population. The results indicate that bacterial buoyant density is inversely related to the amount of capsular polysaccharide enveloping the cell and that a determination of the density profile of the bacteria may be used for discriminating strains with an increased pathogenic potential.     

Impaired phytohaemagglutinin-induced cytotoxicity in vitro of lymphocytes from patients with Hodgkin''s disease or chronic lymphatic leukaemia

Lymphocytes from healthy, unsensitized donors damage allogeneic tissue culture target cells in the presence of phytohaemagglutinin (PHA). In this investigation, blood lymphocytes from eleven patients with clinically active Hodgkin's disease and from eight patients with chronic lymphatic leukaemia (CLL) were tested in vitro for PHA-induced cytotoxicity to Chang cells. Blastoid transformation and DNA-synthesis in the presence of PHA were studied in parallel experiments. The cytotoxicity of the patients' lymphocytes was significantly lower than that of lymphocytes from healthy donors. Synthesis of DNA and blastoid transformation was subnormal in all cases of CLL and in most cases with Hodgkin's disease. A good correlation between cytotoxicity and stimulation by PHA was observed in most cases.

The incidence of positive skin reactions to tuberculin was high in the CLL-group and low in the Hodgkin group. If it is assumed that the pool of immunologically competent cells was normal in the CLL-patients, this would account for their intact cellular reactivity in vivo. In contrast, the high proportion of leukaemic cells could be expected to mask the reactivity of the normal CLL-lymphocytes in vitro. The weak in vitro reactivity of the lymphocytes in Hodgkin's disease is in line with the view that deficient lymphocytes are present in this disease and suggests that the patients' pool of immunologically competent cells is reduced. This could also contribute to the anergic state seen in these patients.

     

The Dominant Epitope of Borrelia garinii Outer Surface Protein C Recognized by Sera from Patients with Neuroborreliosis Has a Surface-Exposed Conserved Structural Motif

Epitope mapping of outer surface protein C (OspC) by using sera from patients with neuroborreliosis led to the identification of one single major immunodominant epitope within the C-terminal 10 amino acid residues. Peptide binding studies and alanine replacement scanning of the C-terminal decapeptide, PVVAESPKKP, revealed a critical role for the PKKP sequence and its terminal carboxyl group for the binding of immunoglobulin M (IgM) antibodies from patients with Lyme borreliosis. Electron microscopy of antibody-labeled spirochetes indicated that the C-terminal region is exposed on the surface of the spirochete. Based on homology to proteins of known function, this region most probably adopts a polyproline II-like helix, which is found in surface-exposed structures involved in protein-protein interactions. This structural motif is highly conserved in Borrelia species causing Lyme borreliosis and subjected to purifying selection. We suggest that the abundance of the C-terminal region of OspC on the surface of B. burgdorferi allows a multimeric high-avidity interaction between the spirochete and surface Igs on B cells. The resulting cross-linking of surface Igs on B cells may induce a T-cell-independent B-cell activation without IgM-to-IgG switching, thus explaining the lack of IgG antibodies to OspC in neuroborreliosis.