Uterine fibroids and longitudinal profiles of the vaginal microbiota in a cohort presenting for transvaginal ultrasound.

Bacterial vaginosis, characterized in part by low levels of vaginal Lactobacillus species, has been associated with pro-inflammatory cytokines which could fuel uterine fibroid development. However, prior work on the associations between uterine fibroids and vaginal bacteria is sparse. Most studies have focused on assessment of individual taxa in a single sample. To address research gaps, we sought to compare short, longitudinal profiles of the vaginal microbiota in uterine fibroid cases versus controls with assessment for hormonal contraceptives (HCs), a possible confounder associated with both protection from fibroid development and increases in Lactobacillus-dominated vaginal microbiota. This is a secondary analysis of 83 reproductive-age cisgender women who presented for transvaginal ultrasound (TVUS) and self-collected mid-vaginal swabs daily for 1-2 weeks before TVUS (Range: 5-16 days, n = 697 samples). Sonography reports detailed uterine fibroid characteristics (N = 21 cases). Vaginal microbiota was assessed by 16S rRNA gene amplicon sequencing and longitudinal microbiota profiles were categorized by hierarchical clustering. We compared longitudinal profiles of the vaginal microbiota among fibroid cases and controls with exact logistic regression. Common indications for TVUS included pelvic mass (34%) and pelvic pain (39%). Fibroid cases tended to be older and report Black race. Cases less often reported HCs versus controls (32% vs. 58%). A larger proportion of cases had low-Lactobacillus longitudinal profiles (48%) than controls (34%). In unadjusted analysis, L. iners-dominated and low-Lactobacillus profiles had higher odds of fibroid case status compared to other Lactobacillus-dominated profiles, however these results were not statistically significant. No association between vaginal microbiota and fibroids was observed after adjusting for race, HC and menstruation. Results were consistent when number of fibroids were considered. There was not a statistically significant association between longitudinal profiles of vaginal microbiota and uterine fibroids after adjustment for common confounders; however, the study was limited by small sample size.

Comparison of Computer-Assisted Self-Interview Versus Clinician Interview for Self-Reported Vulvovaginal Symptoms.

ABSTRACT: This secondary analysis (N = 43) compared computer-assisted self-interview (CASI) to clinician interview for self-report of 8 vulvovaginal symptoms. Concordance was moderate between interview modes (range, 70-86%) with itching and odor having highest agreement. Although prior reports suggest more responses on CASI, we found CASI did not significantly increase self-report of symptoms over clinician interview.

Lactobacillus-dominance and rapid stabilization of vaginal microbiota in combined oral contraceptive pill users examined through a longitudinal cohort study with frequent vaginal sampling over two years.

BACKGROUND: Bacterial vaginosis (BV), a condition in which vaginal Lactobacillus spp. are in low abundance, is associated with vulvovaginal symptoms, obstetric outcomes and urogenital infections. Recurrent BV is difficult to manage, and emerging data indicate a reduced risk of BV with the use of hormonal contraception (HC). Despite widespread use, little longitudinal data is available on whether, and in what timeframe, combined oral contraceptive pills (COCs) may act to affect vaginal microbiota stability and Lactobacillus dominance. METHODS: We compared the vaginal microbiota of reproductive-age cisgender women during intervals on combined estrogen and progestin COCs with non-use intervals in a 2-year observational study. Vaginal microbiota were characterized by 16S rRNA gene amplicon sequencing. FINDINGS: COC users were more likely to have Lactobacillus-dominated microbiota and more stable microbiota over time. Stability increased and then plateaued four weeks after COC initiation. The associations between COCs and Lactobacillus spp. dominance, and microbiota stability, were statistically significant for White, but not African American women; however sample size was limited for African American participants. Findings were similar for other forms of HC and when excluding samples collected during menses. INTERPRETATION: Our study provides a methodologic framework to evaluate observational longitudinal microbiota data with exposure crossovers. We found COCs are associated with vaginal microbiota stability and a Lactobacillus-dominated state. COCs appear to impact stability within a month of initiation. Our findings have clinical implications for how soon benefits can be expected in (at least White) patients initiating COCs, and support the need for larger prospective trials to verify our results in ethnically diverse populations. FUNDING: R01-AI089878.

Vaginal cytokine profile and microbiota before and after lubricant use compared with condomless vaginal sex: a preliminary observational study.

BACKGROUND: Limited data suggest that personal lubricants may damage the vaginal mucosal epithelium, alter the vaginal microbiota, and increase inflammation. We compared vaginal cytokine profiles and microbiota before and after vaginal lubricant use and condomless vaginal sex. METHODS: Reproductive-age women were recruited to a 10-week observational cohort study and were asked to self-collect vaginal samples and behavioral diaries daily. This nested case-control analysis utilized samples collected before and after self-reported condomless sexual activity with lubricants (22 case participants) and without lubricants (22 control participants). Controls were matched to cases on race/ethnicity. Microbiota composition was characterized by sequencing amplicons of the 16S rRNA gene V3-V4 regions. Cytokine concentrations were quantified using a magnetic bead 41-plex panel assay and read using a Bio-Plex 200 array reader. Wilcoxon signed-rank tests were used to assess baseline differences in vaginal cytokines between cases and controls as well as differences pre- and post-exposure. Linear mixed effects models were used to examine differences in relative post-to-pre change in each individual cytokine between matched cases and controls. Similar analyses were conducted for the microbiota data. RESULTS: Mean age was 29.8 years (SD 6.8), and 63.6% were African American. There were few statistically significant changes in cytokines or microbiota before and after exposure in cases or controls. In mixed-effects modeling, the mean relative post-to-pre change of cytokines was higher in cases vs. controls for macrophage derived chemokine (MDC) (p = 0.03). The microbiota data revealed no significant changes when measured by similarity scores, diversity indexes and descriptive community state types (CST) transition analyses. However, post sexual activity, the mean relative abundance of L. crispatus decreased for those who used lubricants (particularly those who were L. iners-dominated prior to exposure). CONCLUSIONS: Although there were overall few differences in the vaginal microbiota and cytokine profiles of lubricant users and controls before and after condomless vaginal sex, there was a trend toward decreases in relative abundance of L. crispatus following use of lubricant. Future larger studies that take into account osmolarity and composition of lubricants may provide additional insights.

Observational cohort study of the effect of a single lubricant exposure during transvaginal ultrasound on cell-shedding from the vaginal epithelium.

The outer layers of the vaginal epithelium (VE) are important because they accumulate glycogen which, under optimal conditions, Lactobacillus spp. consume to grow and acidify the vaginal microenvironment with lactic acid. We hypothesized that exposure to lubricant, for example in the conduct of a transvaginal ultrasound (TVUS), may contribute to the shedding of mature epithelial cells, exposing immature cells. Cervicovaginal fluid (CVF) was sampled at four time points by menstrual cup (Softdisc™) from 50 women referred for TVUS, during which a controlled volume of lubricant was applied to the TVUS wand. Samples were collected (1) immediately before TVUS and (2) 6-12 hours, (3) within one week, and (4) two weeks after TVUS. Clinical vaginal lubricants are similar to commercial lubricants, and often have a high osmolality or pH, and contain bactericides such as methylparaben and propylparaben. The number and maturity of epithelial cells in each CVF sample were measured by quantitative and differential fluorimetry (maturity index, MI). Comparisons of cell-counts and maturity were made by paired Wilcoxon signed-rank tests. Among women with a high pre-TVUS MI (> 3), there was a decrease in median cell-count and mean MI in the sample collected 6-12 hours after TVUS (p<0.001, n = 26 and p < 0.001, n = 26, respectively). For these women, cell-count and MI remained lower in the sample collected within the subsequent week (p<0.001, n = 29 and p<0.01, n = 29, respectively), and MI remained lower in the sample collected within two weeks of TVUS (p<0.01, n = 25), compared to the pre-TVUS sample. Among participants with a low pre-TVUS MI (< 3), cell-count was higher in the sample collected within two weeks of TVUS compared to the pre-TVUS sample (p = 0.03, n = 15), but no significant changes in MI were observed. Results were similar when restricted to reproductive-age women. This preliminary data indicates hypertonic vaginal lubricants may increase vaginal epithelial cell shedding.

Clustering of hypertension, diabetes and overweight/obesity according to socioeconomic status among Bangladeshi adults.

The prevalences of hypertension, diabetes and overweight/obesity are increasing in most developing countries, including Bangladesh. Although earlier studies have investigated the factors associated with these three conditions, little is known about whether socioeconomic status is associated with their co-existence. This cross-sectional study analysed data from the 2011 Bangladesh Demographic and Health Survey. An individual was considered hypertensive, diabetic and overweight/obese if their systolic/diastolic blood pressure, fasting plasma glucose concentration, and body mass index were ≥130/80 mmHg, ≥7 mmol/l and ≥23 kg/m2, respectively. Furthermore, individuals who reported taking anti-hypertensive and anti-diabetic drugs were also considered as hypertensive and diabetic, respectively. Two socioeconomic variables were investigated: education level and household wealth quintile. Descriptive analyses and multilevel logistic regression were conducted. Among the 7932 respondents (50.5% female) aged ≥35 years, the prevalences of hypertension, diabetes, overweight/obesity, any one condition and the co-existence of the three conditions were 48.0%, 11.0%, 25.3%, 60.9% and 3.6%, respectively. In adjusted analysis, individuals with secondary (adjusted odds ratio [AOR]: 1.8, 95% confidence interval [CI]: 1.2-2.8) and college or above (AOR: 3.6; 95% CI: 2.2-5.7) education levels had higher odds of the co-existence of all three conditions compared with those with no formal education. Similarly, compared with the poorest wealth quintile, the richer (AOR: 4.6; 95% CI: 2.2-9.4) and richest (AOR: 11.8; 95% CI: 5.8-24.1) wealth quintiles had higher odds of co-existence of these three conditions. Education and wealth quintile also showed significant relationships with each of the three conditions separately. In conclusion, in Bangladesh, hypertension, diabetes and overweight/obesity are associated with indicators of higher socioeconomic status. These findings highlight the importance of developing healthy lifestyle interventions (e.g. physical exercise and dietary modification) targeting individuals of higher socioeconomic status to minimize the burden of these non-communicable diseases.

Establishment of a non-human primate model for menopausal hot flushes.

Menopause affects the quality of life of millions of women. With modern lifespan the postmenopausal attenuation of circulating estrogen levels can negatively impact a women's life for 30-40 years. The major hypoestrogenic consequence is hot flushes but decline in cognitive function, sleep disorders, depression/anxiety, cardiovascular disease, and osteoporosis are also characteristic for the menopause. Current treatments of hot flushes include estrogen therapy alone or in combination with progestins, soy products, and serotonin and norepinephrine reuptake inhibitors. However, with the exception of estrogens, none of these have satisfactory efficacy. But estrogens come with the unwanted side effects in the periphery, including stimulation of the uterus and breast leading to elevated cancer risk. Therefore, a tremendous effort has been devoted to developing safer therapies and the research has utilized classic rodent models of hot flush with considerable limitations. As hot flushes are primate-specific symptoms, the development of a non-invasive primate hot flush model would have a tremendous impact on drug development. Therefore, our aim was to develop such a non-human primate (NHP) model a hot flush that both recapitulates flushes women experience and is minimally invasive. We investigated if recent developments in thermal imaging have made it possible to accurately monitor skin temperature via camera imaging. In this study, the skin temperature of an ovariectomized rhesus monkey was measured continuously with an infrared camera in a freely moving animal over long time period. Following mapping skin temperatures of several areas of the neck and face we found that the nose of the monkeys showed that largest changes in skin temperature. In the ovariectomized monkey the temperature of the skin on the nose shows up to 9 °C elevations representing hot flushes. In the untreated monkey, hot flushes occurred more frequently in late afternoon/early evening hours than in the morning and last for several minutes. We observed 58 flushes in the 64 evenings of observation. The average number of hot flushes was 0.51 per evening. Oral administration of biotin (niacin) for seven days exaggerated the number of hot flushes to 2.43 per evening. Oral treatment with estradiol benzoate prevented hot flushes and only 2 flushes were detected in the 12 evenings after treatment, averaging 0.17 per evening. The development of this NHP model of hot flush provides great hope for utilizing it for future drug development and mechanistic studied.

A cross-sectional pilot study of birth mode and vaginal microbiota in reproductive-age women.

Recent studies suggest that birth mode (Cesarean section [C-section] or vaginal delivery) is an important event in the initial colonization of the human microbiome and may be associated with long-term health outcomes. We sought to determine the association between a woman's birth mode and her vaginal microbiota in adulthood. We re-contacted 144 adult women from two U.S. studies and administered a brief survey. Vaginal microbiota was characterized on a single sample by amplicon sequencing of the V3-V4 hypervariable regions of the 16S rRNA gene and clustered into community state types (CSTs). We evaluated the association between birth mode and a CST with low relative abundance of Lactobacillus spp. ("molecular bacterial vaginosis" [Molecular-BV]) compared to Lactobacillus-dominated CSTs in logistic regression modeling which adjusted for body mass index, a confounder in this analysis. Twenty-seven women (19%) reported C-section. Overall, C-section showed a non-significant trend towards increased odds of Molecular-BV (aOR = 1.22, 95% CI: 0.45, 3.32), and Prevotella bivia was the strongest single taxa associated with C-section. However, because the two archived studies had different inclusion criteria (interaction p = 0.048), we stratified the analysis by study site. In the study with a larger sample size (n = 88), women born by C-section had 3-fold higher odds of Molecular-BV compared to vaginally-delivered women (aOR = 3.55, p = 0.06, 95% CI: 0.97-13.02). No association was found in the smaller study (n = 56, aOR = 0.19, p = 0.14, 95% CI: 0.02-1.71). This pilot cross-sectional study suggests a possible association between C-section and Molecular-BV in adulthood. However, the analysis is limited by small sample size and lack of comparability in participant age and other characteristics between the study sites. Future longitudinal studies could recruit larger samples of women, address the temporal dynamics of vaginal microbiota, and explore other confounders, including maternal factors, breastfeeding history, and socioeconomic status, which may affect the relationship between birth mode and vaginal microbiota.

Bacterial vaginosis diagnosis and treatment in postmenopausal women: a survey of clinician practices.

OBJECTIVE: Some diagnostic features of the genitourinary syndrome of menopause (GSM) and bacterial vaginosis (BV) overlap, such as low levels of vaginal Lactobacillus and pH > 5. We sought to determine clinicians' diagnostic and treatment practices for postmenopausal women presenting with BV and GSM scenarios and how commercial molecular screening tests are utilized. METHODS: Anonymous surveys were sent to practicing women's health clinicians to evaluate assessment and treatment strategies for postmenopausal women presenting with BV and GSM scenarios. RESULTS: When given a scenario of a postmenopausal woman with symptoms overtly positive for BV, a majority of providers (73%) would conduct a wet mount, though only 35% would evaluate full Amsel's criteria. A majority (89%) recommended treatment with antibiotics, 28.2% recommended vaginal estrogen in addition to antibiotics, and 11.8% recommended vaginal estrogen alone. Of providers who would use a molecular swab, 30% would wait for results before treating the patient's symptoms. When given a scenario of a postmenopausal woman presenting with GSM, a majority (80%) recommended vaginal estrogen, and only 4.6% recommended antibiotics. Few (16%) responders would evaluate with a molecular swab, half of whom would wait for results before prescribing treatment. Clinicians in practice for less than 10 years were more likely to rely on molecular swabs than those who had been practicing longer (P < 0.0003). CONCLUSIONS: Methods used to evaluate postmenopausal women with vaginal symptoms vary. Future studies of postmenopausal women that differentiate diagnostic criteria between BV and GSM, and validate commercial molecular testing for BV in women over age 50 are needed.

Effect of Extended Prophylactic Antibiotic Duration in the Treatment of Open Fracture Wounds Differs by Level of Contamination.

OBJECTIVE: To determine the association between prophylactic antibiotic duration after the definitive wound closure of an open fracture and deep surgical site infection (SSI). DESIGN: Retrospective cohort study. SETTING: 41 clinical sites in the United States, Canada, Australia, Norway, and India. PARTICIPANTS: Patients (N = 2400) with open fractures of the extremities who participated in the Fluid Lavage of Open Wounds (FLOW) trial. INTERVENTION: Extended antibiotic prophylaxis, defined as more than 72 hours of continuous antibiotic use after definitive wound closure. MAIN OUTCOME MEASUREMENT: Deep SSI diagnosed within 1 year of enrollment. RESULTS: Forty-two percent of participants received extended antibiotic prophylaxis. Deep SSI prevalence was 5%, 8%, and 23% for wounds with mild, moderate, and severe contamination, respectively. In open fractures with mild contamination, extended antibiotic use showed a trend toward increased odds [adjusted odds ratio (aOR) = 1.39; 95% confidence interval (CI), 0.92-2.11] of deep SSI compared with shorter use. No association was found among patients with moderate contamination (aOR = 1.09; 95% CI, 0.53-2.27). By contrast, extended antibiotic prophylaxis was strongly protective (aOR = 0.20; 95% CI, 0.07-0.60) against deep SSI in patients with severe contamination. Propensity score sensitivity analysis results were consistent with these findings. CONCLUSIONS: The evidence suggests differential effects of extended postclosure antibiotic duration on SSI odds contingent on the degree of contamination in open fracture wounds. Although extended antibiotic duration resulted in lower odds of SSI among patients with severely contaminated wounds, we observed a trend toward higher odds of SSI in mildly contaminated wounds. LEVEL OF EVIDENCE: Therapeutic Level III. See Instructions for Authors for a complete description of levels of evidence.

Alcohol consumption and serum metabolite concentrations in young women.

PURPOSE: Alcohol consumption is an established breast cancer risk factor, though further research is needed to advance our understanding of the mechanism underlying the association. We used global metabolomics profiling to identify serum metabolites and metabolic pathways that could potentially mediate the alcohol-breast cancer association. METHODS: A cross-sectional analysis of reported alcohol consumption and serum metabolite concentrations was conducted among 211 healthy women 25-29 years old who participated in the Dietary Intervention Study in Children 2006 Follow-Up Study (DISC06). Alcohol-metabolite associations were evaluated using multivariable linear mixed-effects regression. RESULTS: Alcohol was significantly (FDR p < 0.05) associated with several serum metabolites after adjustment for diet composition and other potential confounders. The amino acid sarcosine, the omega-3 fatty acid eicosapentaenoate, and the steroid 4-androsten-3beta,17beta-diol monosulfate were positively associated with alcohol intake, while the gamma-tocopherol metabolite gamma-carboxyethyl hydroxychroman (CEHC) was inversely associated. Positive associations of alcohol with 2-methylcitrate and 4-androsten-3beta,17beta-diol disulfate were borderline significant (FDR p < 0.10). Metabolite set enrichment analysis identified steroids and the glycine pathway as having more members associated with alcohol consumption than expected by chance. CONCLUSIONS: Most of the metabolites associated with alcohol in the current analysis participate in pathways hypothesized to mediate the alcohol-breast cancer association including hormonal, one-carbon metabolism, and oxidative stress pathways, but they could also affect risk via alternative pathways. Independent replication of alcohol-metabolite associations and prospective evaluation of confirmed associations with breast cancer risk are needed.

Patterns in On-time, Daily Submission of a Short Web-Based Personal Behavior Survey in a Longitudinal Women's Health Study.

We evaluated compliance with submitting a short Web-based personal behavior survey daily during a 10-week study (n = 52 women/3419 diaries). Time-stamped forms revealed that 50% of diaries were submitted within 24 hours of the email prompt, and 19% were missing or submitted more than 3 days late. Late submissions may affect data quality.