Characterization of GCY1 in Saccharomyces cerevisiae by metabolic profiling.

AIMS: The analytical study of intracellular (IC) metabolites has developed with advances in chromatography-linked mass spectrometry and fast sampling procedures. We applied the IC metabolite analysis to characterize the role of GCY1 in the glycerol (GLY) catabolic pathway in Saccharomyces cerevisiae. METHODS AND RESULTS: Strains with disrupted or overexpressing GLY catabolic genes such as GCY1, DAK1 and DAK2 were constructed. The strains were cultivated under different aeration conditions and quickly quenched using a novel rapid sampling port. IC concentrations of GLY, dihydroxyacetone (DHA), glycerol 3-phosphate and dihydroxyacetone phosphate were analysed in the strains by gas chromatography/mass spectrometry. DHA was not detected in the gcy1 gene-disrupted strain but accumulated 225.91 µmol g DCW(-1) in a DHA kinase gene-deficient strain under micro-aerobic conditions. Additionally, a 16.1% increase in DHA occurred by overexpressing GCY1 in the DHA kinase-deficient strain. CONCLUSIONS: Metabolic profiling showed that the GCY1 gene product functions as a GLY dehydrogenase in S. cerevisiae, particularly under micro-aerobic conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Metabolic profiling of the GLY dissimilation pathway was successfully demonstrated in S. cerevisiae, and the function of GCY1 was explained by the results.

Monitoring std prevalence and reproductive health care among high-risk adolescent women

Background: Urban adolescent women, particularly those in socially disadvantaged situations are at high risk for infections with Chlamydia trachomatis (CT), and Neisseria gonorrhoeae (GC) and their sequelae. Non-invasive screening technologies, including ligase chain reaction (LCR) and polymerase chain reaction (PCR) urine tests for CT and GC allow for provision of screening services in settings frequented by high-risk adolescents where such services have not been traditionally provided. In 1999, the CDC lead a multi-site project monitoring STD prevalence and reproductive health service, collecting a standard set of variables in women <20 yrs in juvenile justice system, drug treatment centers, and school-based clinics. This report provides preliminary data from Alabama and Colorado. Methods: Adolescent women who present for intake/services at selected facilities are enrolled in the project. In collaboration with CDC, sites developed a set of standard variables to be collected, including demographics, risk behaviors, symptoms, test performed, and test results. Dependent on services already performed, sites added CT and/or GC urine PCR/LCR and pregnancy testing to existing screening protocols.Results: In first 3-months, 533 adolescent women were enrolled in this on-going study; 379 in juvenile institutions, 38 in drug treatment centers, and 116 in school-based clinics. Mean age was 16 yrs, over 90% were sexually experienced, nearly 50% did not use condom with the last sex, less than 40% reported "current" birth control use, over 2/3 had sexual intercourse in past 3 months, many with multiple partners. Prevalence rates for CT and GC are summarized in the table. Conclusions: Non-invasive (urine) LCR/PCR tests allow for easy monitoring of selected STDs among adolescent women in high-risk settings. These findings may contribute to a sentinel surveillance system among high-risk populations and, in turn, may provide the rationale for the development of health care/intervention priorities at policy level.

Population-level intervention to promote chlamydia screening. Moving toward implementation of chlamydia hedis 2000 measure

Background: HEDIS 2000 measure includes chlamydia screening in women which is designed to assess the percentage of sexually active women 15 to 25 years who have received at least one screening test for chlamydia during the reporting year. This study is being undertaken to determine feasibility of implementing a population-level intervention within HMOs to promote chlamydia screening. This abstract presents preliminary findings from the Birmingham project of this multicenter study.Methods: In partnerships with two HMOs, series of outreach methods were used in a stepwise fashion to determine potential barriers and enabling factors for the implementation of chlamydia HEDIS measure in a conservative social environment. Mail outreach was sequentially combined with newspaper, TV, radio advertisements and poster displays. Both qualitative and quantitative impact of the outreach efforts were measured across the timeline. The measures included reporting for chlamydia screening (urine LCR) and infection rate, monitoring chlamydia hotline and staffed phoneline use, and assessment of untoward effects and cost-analysis of the chlamydia outreach campaign.Results: The key findings are: the benefit of chlamydia screening is not understood by general public, letters send by Health Plans to their members are not read by many subscribers, and there are wide gaps between adolescents and their parents, in knowledge, attitudes, beliefs in regard to obtaining information and accessing the screening services (teens prefer hotline, brochure in an envelop addressed to teens, incentives for reporting to the clinic for screening, vs. parents prefer staffed phone consults, "exposed" brochure addressed to parents, and no incentives). A month of sustained and repeat multi-media campaign resulted in 330 hotline calls, 83 phone calls and only 17 subjects being tested (3 were positive) though many more intended to come. Cumulative effects and cost of various outreach efforts are being monitored. Informational chlamydia brochures and free test cards mailed to the homes generated no negative response from the subscribers, contrary to the concerns of the HMOs. Conclusion: To be effective, investment in public education campaign and social marketing strategies must be integrated in population-level intervention for the implementation of the chlamydia HEDIS 2000 measure.