TSPY4 is a novel sperm-specific biomarker of semen exposure in human cervicovaginal fluids; potential use in HIV prevention and contraception studies

Background

Developing an objective, reliable method to determine semen exposure in cervicovaginal fluids is important for accurately studying the efficacy of vaginal microbicides and contraceptives. Y-chromosome biomarkers offer better stability, sensitivity, and specificity than protein biomarkers. TSPY4 belongs to the TSPY (testis-specific protein Y-encoded) family of homologous genes on the Y-chromosome. Using a multiplex PCR amplifying TSPY4, amelogenin, and Sex-determining region in the Y chromosome (SRY), our objective was to determine whether a gene in the TSPY family was a more sensitive marker of semen exposure in cervicovaginal fluids than SRY.

Study Design

The multiplex polymerase chain reaction (PCR) was developed using sperm and vaginal epithelial (female) DNA. Diluted sperm DNA and mixed male/female DNA was used to determine the sensitivity of the multiplex PCR. Potential interference of TSPY4 amplification by components in cervicovaginal and seminal fluids was determined. TSPY4 and SRY amplification was also investigated in women participating in a separate IRB-approved clinical study in which cervicovaginal swab DNA was collected before semen exposure and at various time points after exposure.

Results

TSPY4, SRY, and amelogenin were amplified in sperm DNA, but only amelogenin in female DNA. The limit of sperm DNA from which TSPY4 could be amplified was lower than SRY (4 pg vs 80 pg). TSPY4 could also be amplified from mixed male/female DNA. Amplification was not affected by cervicovaginal and seminal components. Using cervicovaginal swab DNA from three women before and after semen exposure, TSPY4 was detected up to 72 h post exposure while SRY detection was observed up to 24–48 h. TSPY4 was detected up to 7 days post exposure in one out of three women.

Conclusions

We have demonstrated that TSPY4 is a new sensitive, and sperm-specific biomarker. The multiplex PCR incorporating this new biomarker has potential to be an objective measure for determining semen exposure in clinical trials of vaginal products such as contraceptives and HIV pre/post-exposure prophylaxis agents.     

Pharmacokinetics and adhesion of the Agile transdermal contraceptive patch (AG200-15) during daily exposure to external conditions of heat,humidity and exercise

BackgroundThis study compares the pharmacokinetic profile, adhesion and safety of the AG200-15 Agile Patch (AP), a novel contraceptive patch releasing low-dose ethinyl estradiol (EE) and levonorgestrel (LNG), during wear under external conditions of heat, humidity and exercise versus normal activities.Study DesignThis open-label, three-period, five-treatment, crossover study randomized 24 healthy women to one of six external condition sequences. Each sequence included one normal wear and two external conditions periods. Participants wore the AP for 7 days under normal conditions or conditions of daily sauna, treadmill, whirlpool or cool water immersion, with a 7-day washout between treatments. Blood samples were collected for pharmacokinetic evaluations.ResultsTwenty-four subjects completed the study. For EE, the mean maximum concentration level (C_max), area under the plasma concentration–time curve from time 0 to 168 h (AUC_0–168) and area under the plasma concentration–time curve from time 0 to infinity (AUC_0–inf) were higher during normal conditions compared with all external conditions (geometric means ratio range: 80%–93%), except cool water. Mean steady-state concentrations (C_ss) of EE were highest under normal conditions, followed by cool water, sauna, whirlpool and treadmill. The LNG mean C_max, AUC_0–168, AUC_0–inf and C_ss were higher under normal wear versus all other conditions (geometric means ratios: 75%–82%), with the exception of AUC_0–168, AUC_0–inf and C_ss for cold water. Median times to maximum concentration (T_max) for EE and LNG were comparable across conditions. Patch adhesion was excellent under all conditions. Adverse events were mild, with none serious or leading to discontinuation.ConclusionsAlthough slightly lower mean drug concentration levels were observed for whirlpool, treadmill and sauna, drug concentrations under all conditions were well within therapeutic ranges established for the AP during normal wear and within ranges reported for low-dose combination oral contraceptives. Patch adhesion was excellent; the AP was safe and well tolerated under all conditions.     

Actions of platelet activating factor (PAF) homologues and their combinations on neutrophil chemokinesis and cutaneous inflammatory responses in man

The inflammatory actions of synthetic C16:0 and C18:0 platelet activating factor (PAF) homologues, both alone and in combination, have been compared in an in vitro human neutrophil chemokinesis assay and by intradermal injection in human skin. In the chemokinesis assay, the maximum distance moved by neutrophils in the presence of C18:0 PAF was significantly greater than that seen with the C16:0 compound. A mixture of C16:0 and C18:0 PAFs in a ratio of 1:9 appeared to be more active than in a ratio of 3:1. Intradermal injection of the C16:0 and C18:0 PAF homologues induced dose-dependent increases in weal volume and flare area responses which were not significantly different. Combination of these phospholipids in a ratio of 3:1 or 1:9 of C16:0:C18:0 did not significantly alter the dose response curves. Thus, changes in the chain length of the alkyl substituent of synthetic PAF homologues and combination of these homologues, in ratios found in vivo or formed by leukocytes in vitro, did not alter the cutaneous inflammatory responses to PAF in man. The C18:0 homologue was, however, more active as a human neutrophil chemoattractant in vitro.     

An Internet snapshot study to compare the international availability of the novel psychoactive substance methiopropamine

Context. With the increased use of novel psychoactive substances, there is an increasing availability of these substances from Internet-based suppliers. Methiopropamine, first reported in 2011, is a recreational drug available over the Internet. The aim of this study was to investigate availability and cost of methiopropamine in three different countries: the UK, France, and Canada. Methods. Using the European Monitoring Centre for Drugs and Drug Addiction Internet snapshot methodology, this study, conducted in June 2013, was undertaken in two different languages: in English (the UK and Canada) and in French (France and Canada), using three Internet searching engines: “google.co.uk”, “google.fr” and “google.ca”. Results. A total of 62 sites were found, most of them were found from the English searches. 45% of the suppliers seemed to originate from the UK. The prices of methiopropamine were comparable between suppliers, no matter which search engine or language was used. The cost of a unit of methiopropamine was inversely related to the purchased quantity, going from 19.49 ± 0.15 GBP per gram for a purchase amount of 500 mg to 3.54 ± 0.13 GBP per gram for a purchase amount of 1 kg. Discussion. The results of the present study demonstrate that the sale of methiopropamine has the potential to reach users across the world. It also appears to support that snapshot studies could be used for toxicovigilance across different countries, by studying the Internet market of novel psychoactive substances. Conclusion. To date, snapshot studies, used to monitor the Internet novel psychoactive substances market, have only been undertaken in Europe. We have shown that the flexibility of this methodology enables comparison of the online activity of drug sellers between different countries and continents and that, at least for methiopropamine, the UK is the predominant source for Internet supply.     

Ethinyl estradiol and levonorgestrel pharmacokinetics with a low-dose transdermal contraceptive delivery system, AG200-15: a randomized controlled trial

BackgroundThis study evaluated the ethinyl estradiol (EE) and levonorgestrel (LNG) pharmacokinetic profiles of AG200-15, a transdermal contraceptive delivery system, compared with a combination oral contraceptive (COC) containing EE 35 mcg and norgestimate 250 mcg.Study designA Phase 1, open-label, single-center study in 36 healthy women was conducted over three cycles with a randomized crossover design. After a run-in cycle of 21 days on and 7 days off with AG200-15, participants were randomized to receive one of two treatments: a 21/7-day cycle of AG200-15 either followed or preceded by one cycle of the COC. This trial is registered on ClinicalTrials.gov under the identifier NCT01243580.ResultsDuring the third week of AG200-15 use, mean (±standard deviation) maximum serum concentration (C_max), area under the curve_{0–168 h} and steady-state concentration (C_{ss48–168 h}) for EE were 51.3±17.3 pg/mL, 6.26±2.46 ng h/mL and 35.7±14.5 pg/mL, respectively; for LNG, the corresponding values were 2400±1140 pg/mL, 317±159 ng h/mL and 1847±930 pg/mL, respectively. The AG200-15 EE C_max was approximately 60% lower and the EE C_ss was 15%–20% lower than those obtained with the COC. The calculated daily dose of AG200-15 was equivalent to a 30-mcg EE COC. The most common adverse events (AEs; >10%) in the AG200-15 group were headache, nausea and application-site irritation. All drug-related AEs were mild, and no serious AEs were reported.ConclusionsEE and LNG daily exposure during AG200-15 treatment was within the range reported for a low-dose COC. The daily EE dose with AG 200-15 was equivalent to a 30-mcg COC and was safe and well tolerated.     

Utility of Population Attributable Fraction Assessment in Guiding Interventions to Reduce Low Birthweight in the High-Altitude State of Colorado

Objectives We evaluated the ability of population attributable fraction (PAF) assessments to alter significant modifiable risks for low birthweight (LBW) and the impact of high altitude as a risk for LBW in Colorado. Methods Logistic regression analysis of birth certificate parameters in 1995–1997 identified risk factors for PAF assessment. PAF for birth at high altitude, multiple births, and LBW in singleton births were determined. Subsequent analysis of singleton LBW risks, using number needed to treat (NNT) analysis, estimated how elimination of major modifiable risk factors could reduce LBW in the study population. Public health interventions were initiated and PAF analysis conducted 12 years afterward to determine the effect of interventions. Results PAF in singleton births revealed low maternal weight gain in pregnancy and maternal smoking as the greatest modifiable attributable risk factors for LBW (12.7/12.5 %, respectively, in 1995–1997 and 12.9/7.1 % in 2007–2009). Significant interaction between these variables resulted in PAF of 34.4 % when the two occurred together in 1995–1997, decreasing to 19.4 % in 2007–2009. NNT analysis of singleton births in 1995–1997 revealed that eliminating low maternal weight gain, smoking, late prenatal care in all women and interpregnancy intervals <1 year in multiparous women reduced LBW by 46.5 %. The respective proportional reductions in PAF of 40.3 and 46.3 % for maternal smoking and weight gain/smoking interaction were associated with a 1.4 % LBW reduction in singleton births between the two study periods. Conclusions for Practice PAF and NNT analyses are valuable tools to predict intervention targets to lower LBW.     

A Thematic Analysis of Mothers’ Motivations for Blogging

Introduction Various forms of social media are used by many mothers to maintain social ties and manage the stress associated with their parenting roles and responsibilities. ‘Mommy blogging’ as a specific type of social media usage is a common and growing phenomenon, but little is known about mothers’ blogging-related experiences and how these may contribute to their wellbeing. This exploratory study investigated the blogging-related motivations and goals of Australian mothers. Methods An online survey was emailed to members of an Australian online parenting community. The survey included open-ended questions that invited respondents to discuss their motivations and goals for blogging. A thematic analysis using a grounded approach was used to analyze the qualitative data obtained from 235 mothers. Results Five primary motivations for blogging were identified: developing connections with others, experiencing heightened levels of mental stimulation, achieving self-validation, contributing to the welfare of others, and extending skills and abilities. Discussion These motivations are discussed in terms of their various properties and dimensions to illustrate how these mothers appear to use blogging to enhance their psychological wellbeing.