Chicken vitellogenin gene-binding protein, a leucine zipper transcription factor that binds to an important control element in the chicken vitellogenin II promoter, is related to rat DBP.

We screened a chicken liver cDNA expression library with a probe spanning the distal region of the chicken vitellogenin II (VTGII) gene promoter and isolated clones for a transcription factor that we have named VBP (for vitellogenin gene-binding protein). VBP binds to one of the most important positive elements in the VTGII promoter and appears to play a pivotal role in the estrogen-dependent regulation of this gene. The protein sequence of VBP was deduced from a nearly full length cDNA copy and was found to contain a basic/zipper (bZIP) motif. As expected for a bZIP factor, VBP binds to its target DNA site as a dimer. Moreover, VBP is a stable dimer free in solution. A data base search revealed that VBP is related to rat DBP. However, despite the fact that the basic/hinge regions of VBP and DBP differ at only three amino acid positions, the DBP binding site in the rat albumin promoter is a relatively poor binding site for VBP. Thus, the optimal binding sites for VBP and DBP may be distinct. Similarities between the VBP and DBP leucine zippers are largely confined to only four of the seven helical spokes. Nevertheless, these leucine zippers are functionally compatible and appear to define a novel subfamily. In contrast to the bZIP regions, other portions of VBP and DBP are markedly different, as are the expression profiles for these two genes. In particular, expression of the VBP gene commences early in liver ontogeny and is not subject to circadian control.

Genome-wide gene-based analysis suggests an association between Neuroligin 1 (NLGN1) and post-traumatic stress disorder.

Post-traumatic stress disorder (PTSD) develops in only some people following trauma exposure, but the mechanisms differentially explaining risk versus resilience remain largely unknown. PTSD is heritable but candidate gene studies and genome-wide association studies (GWAS) have identified only a modest number of genes that reliably contribute to PTSD. New gene-based methods may help identify additional genes that increase risk for PTSD development or severity. We applied gene-based testing to GWAS data from the Grady Trauma Project (GTP), a primarily African American cohort, and identified two genes (NLGN1 and ZNRD1-AS1) that associate with PTSD after multiple test correction. Although the top SNP from NLGN1 did not replicate, we observed gene-based replication of NLGN1 with PTSD in the Drakenstein Child Health Study (DCHS) cohort from Cape Town. NLGN1 has previously been associated with autism, and it encodes neuroligin 1, a protein involved in synaptogenesis, learning, and memory. Within the GTP dataset, a single nucleotide polymorphism (SNP), rs6779753, underlying the gene-based association, associated with the intermediate phenotypes of higher startle response and greater functional magnetic resonance imaging activation of the amygdala, orbitofrontal cortex, right thalamus and right fusiform gyrus in response to fearful faces. These findings support a contribution of the NLGN1 gene pathway to the neurobiological underpinnings of PTSD.

Microbiological evaluation of female patients in STD clinics.

A total of 215 women patients attending the STD clinic were evaluated in an attempt to isolate the different microorganisms in sexually transmitted diseases (STD). Mycoplasmas (30.22%), Candida species (20.00%), Trichomonas vaginalis (wet mount study; 15.81%), beta haemolytic streptococci (13.48%), Neisseria gonorrhoeae (9.30%), Staphylococcus aureus (13.95%), inclusion bodies of Chlamydia trachomatis (11.60%) and Gram negative organisms (9.30%) were isolated from these patients. Sera of all patients screened for HBsAg by ELISA showed a carrier rate of 12.5 per cent; 29.8 per cent sera were reactive in the VDRL test at the dilutions varying from 1:4 to 1:64.

PIP: The incidence of 10 sexually transmitted diseases (STDs) in 215 prostitutes attending a STD clinic was determined by culture and other standard techniques. A control group of 100 women attending an outpatient family planning clinic in L.T.M. Medical College, Bombay, matched for age and socioeconomic level was the reference. The organisms assayed, the percent of study group found positive, and the percent of controls found positive were: M. hominis, 11.2 and 0.0%; U. urealyticum, 19.1 and 12.0%; Candida to species level, 20.0 and 70.%; beta-hemolytic streptococci, 13.5 and 3.0%; group B streptococci, 6.7 and 0.0%; T. vaginalis, 15.8 and 3.0%; C. trachomatis (by inclusion bodies), 11.6 and 1.0%; coagulase-positive S. aureus, 13.95 and 5.0%; N. gonorrhoeae, 9.3 and 0.0%; and Gram negative bacteria, 9.3 and 1.0%. The VDRL (Venereal Disease Reference Laboratory) seropositivity was 29.5% in STD patients. HBsAG (hepatitis B surface antigen) were present in 12.5%. Most of the women had vaginitis, cervicitis, vulvitis or warts with secondary infections. These incidence rates were discussed relative to several other Indian surveys.

MTBP suppresses cell migration and filopodia formation by inhibiting ACTN4.

Murine double minute (MDM2) binding protein (MTBP) has been implicated in cancer progression. Here, we demonstrate one mechanism by which MTBP inhibits cancer metastasis. Overexpression of MTBP in human osteosarcoma cell lines lacking wild-type p53 did not alter primary tumor growth in mice, but significantly inhibited metastases. MTBP downregulation increased the migratory potential of MDM2(-/-)p53(-/-) mouse embryonic fibroblasts, suggesting that MTBP inhibited cell migration independently of the Mdm2-p53 pathway. Co-immunoprecipitation and mass spectrometric analysis identified alpha-actinin-4 (ACTN4) as an MTBP-interacting protein. Endogenous MTBP interacted with and partially colocalized with ACTN4. MTBP overexpression inhibited cell migration and filopodia formation mediated by ACTN4. Increased cell migration by MTBP downregulation was inhibited by concomitant downregulation of ACTN4. MTBP also inhibited ACTN4-mediated F-actin bundling. We furthermore demonstrated that nuclear localization of MTBP was dispensable for inhibiting ACTN4-mediated cell migration and filopodia formation. Thus, MTBP suppresses cell migration, at least partially, by inhibiting ACTN4 function. Our study not only provides a mechanism of metastasis suppression by MTBP, but also suggests MTBP as a potential biomarker for cancer progression.

Pharmacovigilance using clinical notes.

With increasing adoption of electronic health records (EHRs), there is an opportunity to use the free-text portion of EHRs for pharmacovigilance. We present novel methods that annotate the unstructured clinical notes and transform them into a deidentified patient-feature matrix encoded using medical terminologies. We demonstrate the use of the resulting high-throughput data for detecting drug-adverse event associations and adverse events associated with drug-drug interactions. We show that these methods flag adverse events early (in most cases before an official alert), allow filtering of spurious signals by adjusting for potential confounding, and compile prevalence information. We argue that analyzing large volumes of free-text clinical notes enables drug safety surveillance using a yet untapped data source. Such data mining can be used for hypothesis generation and for rapid analysis of suspected adverse event risk.

Production of H-Y antibody in the ascites fluid of mouse and localization of the antigen on cells and tissues.

A procedure is described for the production of large amounts of ascites fluid containing specific H-Y antibody. The distribution of H-Y antigen on mouse epididymal spermatozoa, thymocytes, and splenocytes was carried out using this specific antibody in the microcytotoxicity test and ELISA. Employing the indirect immunofluorescent technique, the H-Y antigen was localized on the acrosomal membrane of mouse epididymal and washed ejaculated human spermatozoa and on the entire membrane of mouse splenocytes and thymocytes. Immunohistochemical localization of the antigen in the testicular section indicated its presence in the cytoplasm of Leydig cells and on the membrane of Sertoli cells and sperm heads.

Antibodies to N-acetylglucosamine and heparin in acute and remission phases of rheumatic fever.

The etiology of acute rheumatic fever (ARF) is believed to involve an immunological response to group A streptococcal antigens. Antibodies to group A carbohydrate (A-CHO) have been reported in ARF and rheumatic heart disease patients. As N-acetylglucosamine (GlcNAc) units form the major immunodominant regions in A-CHO antigens, we investigated levels of antibodies to GlcNAc and heparin (with repeated sequences of GlcNAC) in rheumatic fever (RF) patients. Serum samples from 26 acute cases (ARF), 18 remission cases and 17 normal healthy subjects were analyzed for IgG and IgM levels of antibodies to GlcNAc and heparin. High titres of IgG antibodies to heparin and GlcNAc were observed in the ARF group in comparison with controls (p less than 0.0025 and 0.0125 respectively). There was no difference in the levels of IgM antibodies. Remission group demonstrated low titres of IgG to heparin and GlcNAc (p less than 0.01 and 0.0125 respectively) in comparison with ARF group. Heparin antibodies of IgM class was comparatively lower in remission group (p less than 0.005). While the role of these antibodies in different phases of RF needs to be investigated, we conclude that GlcNAc antibodies do not play any role in the pathogenesis of RF or rheumatic heart disease.