Dorothy L. Patton

Influence of the Normal Menstrual Cycle on Vaginal Tissue, Discharge, and Microflora

The objective of this study was to examine genital tissue, vaginal fluid, and vaginal microbial flora at 3 phases of the menstrual cycle in asymptomatic women. Vaginal examinations were performed 3 times in 74 women: at the menstrual phase (days 1-5), the preovulatory phase (days 7-12), and the postovulatory phase (days 19-24). Flora of 50 women without bacterial vaginosis (BV) was analyzed separately from flora of 24 women with BV. The volume of vaginal discharge increased and the amount of cervical mucus decreased over the menstrual cycle. Among subjects without BV, the rate of recovery of any Lactobacillus changed little (range, 82% to 98%; P = .2); however, a small increase occurred in the rate of recovery of heavy (3+ to 4+ semiquantitative) growth of Lactobacillus over the menstrual cycle (P = .04). A linear decrease occurred in the rate of recovery of heavy growth of any non-Lactobacillus species, from 72% at days 1-5 to 40% at days 19-24 (P = .002). A linear decrease also occurred in the rate of recovery of Prevotella species, from 56% on days 1-5 to 28% on days 19-24 (P =. 007), while a small linear increase occurred in the rate of recovery of Bacteroides fragilis (P=.05). Among subjects with BV, the only significant change was an increase in the rate of recovery of Lactobacillus, from 33% at days 1-5 to 54% at days 19-24 (P = .008). Among all subjects, the rate of recovery of heavy growth of Lactobacillus increased over the menstrual cycle and, in contrast, the concentration of non-Lactobacillus species tended to be higher at menses, which is evidence that the vaginal flora becomes less stable at this time.

Depomedroxyprogesterone-induced hypoestrogenism and changes in vaginal flora and epithelium ☆

Objective To identify the effects of depomedroxyprogesterone acetate (DMPA) on vaginal microbial flora and epithelium. Methods Women who desired DMPA for contraception were evaluated before and at 3 and 6 months after initiation of 150-mg DMPA injections every 3 months. At each visit, we assessed genital symptoms, vaginal signs, vaginal micro-flora, and histopathology by vaginal biopsies. Results Among 38 women observed for 6 months, there was significant reduction in mean serum estradiol level (99.9 ± 9.3 pg/mL to 26.6 ± 1.6 pg/mL, P < .001). The number of subjects with any Lactobacillus did not change, but the number with hydrogen peroxide (H2O2)–positive Lactobacillus decreased from 20% before to 12% after 6 months of DMPA (P = .005). The log concentration in colony-forming units per milliliter of vaginal fluid of H2O2–positive Lactobacillus decreased in a linear manner from 4.0 ± 0.6 at baseline to 2.5 ± 0.6 after 6 months of DMPA use (P = .006). The mean number of cell layers in the epithelium was reduced slightly from 28.1 ± 0.7 to 25.9 ± 0.9 (P = .05), epithelial thickness decreased from 1.02 ± 0.04 mm to 0.89 ± 0.05 mm (P = .005), and the glycogen-positive thickness decreased from 0.81 ± 0.04 mm at baseline to 0.66 ± 0.05 after 6 months of DMPA use (P = .005). Conclusion Depomedroxyprogesterone acetate produced a systemic hypoestrogenic state associated with decreased H2O2–positive Lactobacillus colonization and slight thinning of the glycogen vaginal epithelial layer. Such changes possibly compromise the vaginal barrier to infection.

Detection of Chlamydia trachomatis in fallopian tube tissue in women with postinfectious tubal infertility

OBJECTIVE Biopsy tissues from women with postinfectious tubal infertility were studied for the presence of Chlamydia trachomatis. STUDY DESIGN Tubal biopsy specimens from 25 women with postinfectious tubal infertility undergoing laparoscopy for repair of fallopian tubes were evaluated by culture, in situ hybridization. Immunocytochemistry, and transmission electron microscopy for the presence of Chlamydia trachomatis. Serum was also tested for Chlamydia trachomatis antibodies. RESULTS Chlamydia trachomatis was detected in postinfectious tubal biopsy specimens in three of 25 patients by culture, 12 of 24 by in situ hybridization, 15 of 22 by immunoperoxidase stain, and two of 10 by transmission electron microscopy. Serum antibody against Chlamydia trachomatis was detected in 15 of 21 patients. CONCLUSION Chlamydia trachomatis deoxyribonucleic acid or antigens were detected at a high percentage (19/24 women) in the biopsy tissues of the fimbrial and peritubal adhesions by in situ hybridization or immunoperoxidase stain, suggesting a persistent infection in these women even after antibiotic treatment.

Amniotic fluid tumor necrosis factor-α and the risk of respiratory distress syndrome among preterm infants

OBJECTIVE We examined the effect of exposure to amniotic fluid infection and cytokines on the pulmonary outcome of preterm infants. STUDY DESIGN A cohort of 136 preterm infants born to women in preterm labor had amniotic fluid cultures and tumor necrosis factor-alpha assays performed. Amniotic fluid was collected by transabdominal amniocentesis. Outcome measures included respiratory distress syndrome and length of oxygen and ventilator support. Logistic regression and Cox proportional hazards regression adjusted for birth weight and other confounders. RESULTS Respiratory distress syndrome developed in 67 (49%) of 136 infants. Elevated amniotic fluid tumor necrosis factor-alpha levels and amniotic fluid infection were significantly associated with respiratory distress syndrome. This association persisted after adjustment for birth weight. Infants exposed to tumor necrosis factor-alpha remained on supplemental oxygen and assisted ventilation longer and had longer hospital stays compared with nonexposed infants. CONCLUSION Prenatal exposure to tumor necrosis factor-alpha may be a risk factor for respiratory distress syndrome and its complications.

Infectious correlates of HIV-1 shedding in the female upper and lower genital tracts.

Objectives: To determine the effects of vaginal, cervical, and endometrial infections on shedding of HIV-1 RNA in the female genital tract. Design: Cross-sectional. Methods: Antiretroviral-naive women from Nairobi, Kenya with CD4 cell counts ≥ 350 cells/μl had plasma and endocervical wick samples collected for HIV quantification by real-time RNA reverse transcriptase-polymerase chain reaction. Vaginal and cervical Gram stains and endometrial biopsies were obtained. Vaginal Gram stain was used to diagnose bacterial vaginosis and to quantify Lactobacillus levels. Results: Twenty-six of 50 (52%) women had detectable endocervical HIV-1 RNA with a median endocervical viral load of 1760 copies/ml (range: undetectable to 1 030 000 copies/ml). Women with decreased Lactobacillus had 15.8-fold [95% confidenceinterval (CI), 2.0–123] greater endocervical HIV-1 RNA than women with normal Lactobacillus levels. Women with plasma cell (PC) endometritis [≥ 1 PC/high-power field (hpf)] had a 15.8-fold (95% CI, 2.0–120) higher endocervical HIV RNA level than women without PC endometritis. Both these associations remained after controlling for plasma viral load. Cervicitis (≥ 30 polymorphonuclear leukocytes/hpf), however, was not associated with endocervical HIV-1 RNA shedding (P = 0.81). Conclusions: In HIV-1-infected, antiretroviral-naive women without symptoms of pelvic inflammatory disease infection, abnormal vaginal flora and inflammatory cells in the endometrium affected HIV-1 shedding from the lower genital tract. These data suggest that both the upper and lower genital tracts contribute to female HIV-1 genital shedding.

Detection of Chlamydia trachomatis deoxyribonucleic acid in women with tubal infertility

Objective To determine the presence of Chlamydia trachomatis by in situ hybridization in fallopian tube tissues of women with distal tubal occlusion. Patients Subjects were selected from a Seattle-based study of infertility in women with distally occluded fallopian tubes undergoing reconstructive surgery. For comparison, six specimens were obtained from women undergoing surgery for noninfertility-related conditions who had normal appearing tubes. Methods Tissue specimens from 16 of these patients were selected for analysis by in situ hybridization and immunoperoxidase (IP) staining. Results C . trachomatis was detected in 9 of 16 women by either in situ hybridization or IP. Six of the nine were seropositive with titers ranging from 1:16 to 1:64. Tissue from 6 women with normal appearing fallopian tubes were negative by in situ hybridization. Conclusions The demonstration of C . trachomatis deoxyribonucleic acid and/or antigens in fallopian tube tissue from infertile women with distal tubal disease suggests that C. trachomatis can persist in these tissues in an uncultivable state.

Preclinical safety and efficacy assessments of dendrimer-based (SPL7013) microbicide gel formulations in a nonhuman primate model

ABSTRACT Three gel formulations (1%, 3%, and 5% [wt/wt]) of SPL7013, a dendrimer known to have antiviral (anti-human immunodeficiency virus and anti-herpes simplex virus) activities, completed a range of preclinical tests in the pigtailed macaque models for vaginally and rectally applied topical microbicide safety assessments. The vaginal safety profile of the 3% SPL7013 gel formulation was equal to that of the 1% formulation but was superior to that of the 5% formulation. The 3% SPL7013 gel was further evaluated for rectal safety and for antichlamydial efficacy with cervical challenge with Chlamydia trachomatis. This first-generation dendrimer-based product was shown to be safe to the vaginal and rectal microenvironments with repeated daily use. However, a single intravaginal application of the 3% (wt/wt) SPL7013 gel did not provide protection from the acquisition of cervical chlamydial infection.

Preclinical Safety Assessments of UC781 Anti-Human Immunodeficiency Virus Topical Microbicide Formulations

ABSTRACT The nonnucleoside reverse transcriptase inhibitor UC781 is under development as a potential microbicide to prevent sexual transmission of human immunodeficiency virus type 1 (HIV-1). Two gel formulations of UC781 (0.1% and 1.0%) were evaluated in a range of preclinical safety assessments, including systemic absorption analysis following topical application in the pig-tailed macaque models for vaginally and rectally applied topical microbicides. High-sensitivity high-performance liquid chromatography analysis of serum samples showed that no systemic absorption of UC781 was detected after repeated vaginal or rectal application of either product. However, high levels of UC781 were detectable in the cervicovaginal lavage samples up to 6 h after product exposure. Both formulations were safe to the vaginal microenvironment, even with repeated daily use, as evidenced by colposcopy, cytokine analysis, and lack of impact on vaginal microflora. By contrast, rectal application of the 1.0% UC781 formulation caused an increased expression of numerous cytokines not observed after rectal application of the 0.1% UC781 formulation. These results provide additional support for the continued development of UC781 formulations as anti-HIV microbicides.

Relationship of vaginal bacteria and inflammation with conception and early pregnancy loss following in-vitro fertilization

Objective: The aim of this study was investigate the impact of vaginal flora and vaginal inflammation on conception and early pregnancy loss following in-vitro fertilization (IVF). Methods: We enrolled 91 women who were undergoing IVF. At embryo transfer (ET), all of the women had quantitative vaginal culture, ET catheter-tip culture, and vaginal Gram stain scored for bacterial vaginosis and quantitated for polymorphonuclear leukocytes (PMNs). Conception and early pregnancy loss were compared with culture and Gram stain results. Statistical analyses included the Chi-square test, Fishers exact test and the Mann–Whitney U-test. Results: The overall live birth rate (LBR) was 30% (27/91), and the rate of early pregnancy loss was 34% (14/41). In women with bacterial vaginosis, intermediate flora and normal flora, the conception rates were 30% (3/10), 39% (12/31) and 52% (26/50), respectively (p = 0.06 for trend). Early pregnancy loss occurred in 33% (1/3), 42% (5/12) and 31% (8/26) of women, respectively (p = 0.06, comparing intermediate and normal flora). The vaginal log concentration of hydrogen peroxide-producing lactobacilli was 7.3 ± 1.7 in women with a live birth (n = 27) and 4.9 ± 2.5 in those with early pregnancy loss (n = 14) (p = 0.1). Conclusions: IVF patients with bacterial vaginosis and with a decreased vaginal log concentration of hydrogen peroxide-producing lactobacilli may have decreased conception rates and increased rates of early pregnancy loss. A larger prospective treatment trial designed to evaluate the impact on IVF outcomes of optimizing the vaginal flora prior to IVF may be warranted.

The vaginal microflora of pig-tailed macaques and the effects of chlorhexidine and benzalkonium on this ecosystem.

Background and Objectives: To characterize normal vaginal microflora of pig‐tailed macaques and to evaluate two commonly used intravaginal compounds, chlorhexidine (CHG), a vaginal antiseptic (Surgilube, E. Fougera, Melville, NY), and benzalkonium chloride (BZK) (spermicidal contraceptive) in this monkey model to assess effects on the vaginal microflora. Study Design: Vaginal swabs were collected for microbiologic analysis to characterize normal flora. Subsequently, the vagina was exposed to either CHG or BZK twice at 24‐hour intervals. Results: The vaginal microflora of 26 pig‐tailed macaques was found to be remarkably similar to the vaginal flora of the human with respect to frequency of vaginal colonization by H2O2‐producing lactobacilli, Prevotella species, and several other microorganisms. After two vaginal applications at 24‐hour intervals, CHG had only small effects on the vaginal microflora of five animals. By contrast, BZK applied by the same protocol had profound adverse effects on the lactobacilli and Peptostreptococcus and more transient effects on vaginal Prevotella and viridans streptococci of six animals. Conclusions: These observations demonstrate that the vaginal microflora of the pig‐tailed macaque is a useful model in which to further evaluate newly developed intravaginal contraceptives that may be microbicidal and/or virucidal before widespread intravaginal use in women.