Zika Virus Infection, Reproductive Organ Targeting, and Semen Transmission in the Male Olive Baboon.

Zika virus (ZIKV) infection in pregnant women is a serious threat to the development and viability of the fetus. The primary mode of ZIKV transmission to humans is through mosquito bites, but sexual transmission has also been well documented in humans. However, little is known of the short- and long-term effects of ZIKV infection on the human male reproductive system. This study examines the effects of ZIKV infection on the male reproductive organs and semen and the immune response of the olive baboon (Papio anubis). Nine mature male baboons were infected with ZIKV (French Polynesian strain) subcutaneously. Six animals were euthanized at 41 days, while three animals were euthanized at 10 or 11 days postinfection (dpi). Viremia and clinical evidence of infection were present in all nine baboons. ZIKV RNA was present in the semen of five of nine baboons. ZIKV was present in the testes of two of three males euthanized at 10 or 11 dpi, but in none of six males at 41 dpi. Immunofluorescence of testes suggested the presence of ZIKV in sperm progenitor cells, macrophage penetration of seminiferous tubules, and increased tumor necrosis factor alpha (TNF-α), particularly in vascular walls. These data demonstrate that male olive baboons approximate the male human ZIKV response, including viremia, the adaptive immune response, and persistent ZIKV in semen. Although gross testicular pathology was not seen, the demonstrated breach of the testes-blood barrier and targeting of spermatogenic precursors suggest possible long-term implications in ZIKV-infected primates.IMPORTANCE Zika virus (ZIKV) is an emerging flavivirus spread through mosquitoes and sexual contact. ZIKV infection during pregnancy can lead to severe fetal outcomes, including miscarriage, fetal death, preterm birth, intrauterine growth restriction, and fetal microcephaly, collectively known as congenital Zika syndrome. Therefore, it is important to understand how this virus spreads, as well as the resulting pathogenesis in translational animal models that faithfully mimic ZIKV infection in humans. Such models will contribute to the future development of efficient therapeutics and prevention mechanisms. Through our previous work in olive baboons, we developed a nonhuman primate model that is permissive to ZIKV infection and transfers the virus vertically from mother to fetus, modeling human observations. The present study contributes to understanding of ZIKV infection in male baboon reproductive tissues and begins to elucidate how this may affect fertility, reproductive capacity, and sexual transmission of the virus.

Decreased very long chain polyunsaturated fatty acids in sperm correlates with sperm quantity and quality.

PURPOSE: To determine if levels of very long chain polyunsaturated fatty acids (VLC-PUFA; ≥ 28 carbons;4-6 double bonds) in human sperm correlate with sperm quantity and quality as determined by a complete semen analysis. METHODS: Ejaculates from 70 men underwent a complete semen analysis, which included volume, count, motility, progression, agglutination, viscosity, morphology, and pH. For lipid analysis, sperm were pelleted to remove the semen. Lipids were extracted from the cell pellet and methyl esters of total lipids analyzed by gas chromatography. The sphingolipids were enriched and sphingomyelin (SM) species measured using tandem mass spectrometry. Pair-wise Pearson correlation and linear regression analysis compared percent VLC-PUFA-SM and percent docosahexaenoic acid (DHA) to results from the semen analysis. RESULTS: VLC-PUFA-SM species having 28-34 carbon fatty acids were detected in sperm samples, with 28 and 30 carbon VLC-PUFA as most the abundant. The sum of all VLC-PUFA-SM species comprised 0 to 6.1% of the overall SM pool (mean 2.1%). Pair-wise Pearson analyses showed that lower levels of VLC-PUFA-SM positively correlated with lower total motile count (0.68) and lower total count (0.67). Total VLC-PUFA-SM and mole % DHA (22:6n3) were not strongly correlated (- 0.24). Linear regression analysis confirmed these findings. CONCLUSION: This study revealed a positive correlation between the levels of VLC-PUFA with sperm count and total motile count and suggests that both sperm quality and quantity may depend on the presence of VLC-PUFA. The lack of correlation between VLC-PUFA and DHA suggests that low VLC-PUFA levels do not result from inadequate PUFA precursors.

In high responding patients undergoing an initial IVF cycle, elevated estradiol on the day of hCG has no effect on live birth rate.

BACKGROUND: The impact of elevated estradiol on the day of human chorionic gonadotropin (hCG) administration on in vitro fertilization (IVF) outcomes has been debated for over 25 years. Some investigators have shown a positive effect, others a negative effect; while most have shown no effect. Few studies have expressed their findings based on live birth. This study examined the relationship between estradiol level and other IVF cycle response parameters in relation to pregnancy, with a focus on live births after controlling for embryo quality. METHODS: We performed a retrospective cohort study on 489 patients <40 years old that underwent an initial IVF cycle. Estradiol concentration on the day of hCG was categorized as; low <2000 pg/ml), mid (2001-4000 pg/ml) and high (>4000 pg/ml) to determine how estradiol level on the day of hCG affected response variables during the IVF cycle. We performed a subgroup analysis restricted to patients with good/fair quality embryos transferred (n=428), to control for embryo quality and assessed pregnancy outcome. The association between estradiol and live birth (LB) was then evaluated after identifying and controlling for confounding factors. Multivariate analysis was used to identify significant main effects and interactions in the model. Estradiol levels were also compared in patients having a LB or not (NLB) in both populations. RESULTS: We found that estradiol was significantly related to + hCG, clinical pregnancy rate, age, and most other IVF cycle response variables. After performing the subgroup analysis controlling for embryo quality, we found that LB rates were not different. Only the main effects of average embryo quality at transfer (AEQS), age and transferring two embryos influenced LB. Estradiol levels were also compared in patients having a LB or NLB in both populations and was found to be higher/not different in LB patients. LB rates and AEQS were also not different in a subgroup of patients having an elevated level of estradiol (>4200 pg/ml) on the day of hCG in patients having embryo transfer on day 3 or day 5. CONCLUSIONS: After controlling for embryo quality, elevated estradiol on the day of hCG had no effect on LB.

Validation of the power model of ovarian nongrowing follicle depletion associated with aging in women.

OBJECTIVE: To validate recently proposed models of ovarian nongrowing follicle (NGF) decay associated with aging within the context of an independent data set. DESIGN: Prospective investigation. SETTING: Academic medical center. PATIENT(S): Normal appearing ovaries collected from 52 women (age 28-51 years) undergoing oophorectomy for benign gynecologic indications. INTERVENTION(S): Determining ovarian NGF counts with systematic random sampling rules and a validated fractionator/optical disector technique. The goodness-of-fit of predicted NGF counts based on the power and double Gaussian models and those observed in the validation set was assessed with the calculation of the Akaike information criterion and R(2) values. MAIN OUTCOME MEASURE(S): The goodness-of-fit between observed and expected ovarian NGF counts. RESULT(S): The power model was an excellent fit to the observed data. The average difference between the observed and expected NGF count was 0.161 (95% CI, -0.058, 0.327). In the present study population, the power model was a superior fit to the observed data compared with the double Gaussian model. CONCLUSION(S): This prospective investigation with an independent set of ovarian NGF counts validates the power model as an excellent characterization of the ovarian NGF decline associated with aging.

Ovarian primordial and nongrowing follicle counts according to the Stages of Reproductive Aging Workshop (STRAW) staging system.

OBJECTIVE: The aim of this study was to characterize the ovarian primordial and nongrowing follicle number according to the Stages of Reproductive Aging Workshop (STRAW) staging system as defined by menstrual cycle characteristics. METHODS: Normal ovaries were collected from 63 women (age 26-52 y) undergoing oophorectomy for benign indications. Before surgical operation, each participant completed a detailed questionnaire collecting information regarding menstrual cycle characteristics and was classified by bleeding patterns into STRAW stages -4, -3, -2, and -1. A single ovary was selected for the determination of ovarian primordial and total nongrowing follicle number using a validated fractionator/optical disector method. A subset of the participants (n = 43) underwent transvaginal ultrasound examination for the determination of the ovarian antral follicle count and serum measurements of follicle-stimulating hormone, estradiol, antimüllerian hormone, and inhibin B. All measurements were obtained within 2 weeks of surgical operation, irrespective of cycle day. RESULTS: Significant differences were identified in ovarian primordial (P < 0.0001) and nongrowing follicle (P < 0.0001) counts across the STRAW stages. In post hoc testing, the differences in primordial follicle counts were significant between each of the STRAW stages. Significant differences were also identified in serum levels of antimüllerian hormone, follicle-stimulating hormone, and ovarian antral follicle count across the STRAW stages. CONCLUSIONS: Progression through the STRAW stages as defined by menstrual cycle characteristics is associated with progressive and significant decreases in the ovarian primordial follicle number.