Human reproductive system microbiomes exhibited significantly different heterogeneity scaling with gut microbiome,but the intra-system scaling is invariant

Maintaining sexual reproduction in a highly competitive world is still one of the major mysteries of biology given the apparently high efficiency of asexual reproduction. Co-evolutionary theories such as the Red Queen hypothesis would suggest that the microbiomes in human reproductive systems, specifically the microbiomes contained in semen and vaginal fluids, should reach some level of homogeneity thanks to arguably the most conspicuous microbiome transmission between two sexes. The long-term sexual coevolution should favor the dynamic homogeneity or stability, which should also be beneficial for sexual reproduction such as sperm survival or fertilization on physiological/ecological time scale. We present a piece of quantitative evidence in the form of microbial community spatial heterogeneity to support the stability notion by analyzing three big datasets of the human vaginal, semen and gut microbiome. Methodologically, we applied a recent community-level extension to the classic Taylor's power law, which reached the rare status of ecological law and has found applications beyond biology. Both ecological and evolutionary theories, such as hologenome/holobiont and Red Queen, as well as consideration of first principles, would predict that microbiome transmissions between two sexes should have homogenizing effects on the composition and stability of the microbiomes in human reproductive systems, and therefore have similar variance structures. This is supported by the finding that the power law analysis revealed human vaginal and semen microbiomes exhibited the same scaling parameter size in their community spatial (inter-individual) heterogeneities, while the both exhibited significantly different heterogeneity scaling parameter size with the human gut microbiome.     

A simplified touch tape preparation from tube cultures for microscopic examination of filamentous fungi

Cellophane touch tape preparation provides reproducible results in minimal time when compared to tease mount and slide culture techniques for the identification of fungi from culture plates, but it is difficult to perform from tube cultures. Here, we describe an easy to perform touch tape preparation method that provided a better result from fungal tube culture.     

Vaginal vault cytology tests: analysis of a decade of data from a UK tertiary centre

H. Stokes‐Lampard, S. Wilson, C. Waddell and L. Bentley Vaginal vault cytology tests: analysis of a decade of data from a UK tertiary centre Objectives: To examine temporal trends in the use of vault cytology tests in primary and secondary care and the demographics of those women tested. Methods: Retrospective analysis of routinely collected data concerning women who had a vault cytology test processed during a 10‐year period (1 April 1995 to 31 March 2005) at Birmingham Women’s NHS Foundation Trust. Results: A total of 8457 vault cytology tests from 3164 women (range 1–17 tests, median = 2) were processed, representing approximately 2% of the cervical cytology workload of the Department of Cytopathology at Birmingham Women's Hospital. There was a significant reduction in annual numbers processed (Pearson correlation ?0.958, P < 0.001). Significant abnormalities (mild dyskaryosis or worse) were detected in 4.5%, with malignancy being detected in <0.1%. The unsatisfactory cytology test rate was 10.7% overall. There was a reduction in the numbers of vault cytology tests coming from the community, hospital outpatient clinics and operating theatres over time (χ² for linear trend = 139.53, 9 d.f., P < 0.0001). Tests originating from community settings had the lowest disease detection rates: no malignancies and only two severe abnormalities were detected from almost 4000 primary care samples; abnormal results represented 2.8% (n = 113), of which the majority (n = 73) were borderline results. All cancers (n = 8) were detected in samples taken in gynaecology and colposcopy clinics. Conclusions: Vault cytology test usage appears to be reducing, particularly from outpatient clinics and primary care. Community detection rates are very low. Further research is required to establish the true costs and benefits of vaginal vault cytology.     

Reproductive tract defense and disease in postpartum dairy cows

This paper briefly reviews recent data and concepts on the development and mitigation of infection and inflammation in the reproductive tract of dairy cows during the first 2 mo after calving. The incidence of metritis is typically between 10 and 20%, of clinical endometritis or purulent vaginal discharge (PVD) approximately 15%, and of subclinical or cytological endometritis a further 15%. Worse postpartum negative energy balance is associated with more severe or prolonged uterine inflammation. Changes in feed intake, expression of genes for pro-inflammatory cytokines, notably interleukin (IL) 1, IL6 and IL8, circulating concentrations of beta-hydroxybutyrate (BHBA) or nonesterified fatty acids (NEFA), and innate immune function precede both metritis and endometritis by several weeks. Infections with Escherichia coli and Arcanobacterium pyogenes are associated with both metritis and PVD. There are new data to suggest that specific virulence factors in E. coli associated with adherence may be important in metritis and PVD. Cytological endometritis and PVD are overlapping but largely distinct conditions, and there are emerging data that cervicitis exists both concurrent with and separate from endometritis. Much remains to be learned about what initiates and sustains harmful inflammation of the reproductive tract. Such information is necessary to develop effective treatments for the various forms of disease and, more importantly, to develop means to prevent endometritis and cervicitis. In particular, vaccination against specific uterine pathogens and interventions to modulate innate immune response appear to be important avenues for investigation. Presently, commonly recommended best management practices for cows in the transition period are likely to be helpful to mitigate the risk of reproductive disease.     

Vaginal extracellular vesicles impair fertility in endometriosis by favoring Th17/Treg imbalance and inhibiting sperm activity

Extracellular vesicles (EVs) play a key role in various diseases. However, their effect on endometriosis (EMs)-associated infertility is poorly understood. We co-cultured EVs from the female vaginal secretions with human sperm and also generated a mouse model of EMs by allogenic transplant to explore the effect of EVs on fertility. EVs from individuals with EMs-associated infertility (E-EVs) significantly inhibited the total motility (26.46% vs. 47.1%), progressive motility (18.78% vs. 41.06%), linear velocity (21.98 vs. 41.91 µm/s) and the acrosome reaction (AR) rate (5% vs. 22.3%) of human sperm in contrast to the control group (PBS). Furthermore, E-EVs dose-dependently decreased the intracellular Ca²⁺ ([Ca²⁺]_i), a pivotal regulator of sperm function. Conversely, healthy women (H-EVs) increased human sperm motion parameters, the AR rate, and sperm [Ca²⁺]_i. Importantly, the mouse model of EMs confirmed that E-EVs further decreased the conception rate and the mean number of embryo implantations (7.6 ± 3.06 vs. 4.5 ± 3.21) compared with the control mice by inducing the production of inflammatory cytokines leading to a Th17/Treg imbalance. H-EVs could restore impaired fertility by restoring the Th17/Treg balance. We determined the impact of EVs derived from the female genital tract on human sperm function and studied the possible mechanisms by which it affects fertility. Our findings provide a novel rationale to ameliorate EMs-associated infertility.     

Novel vaginal drug delivery system: deformable propylene glycol liposomes-in-hydrogel

Deformable propylene glycol-containing liposomes (DPGLs) incorporating metronidazole or clotrimazole were prepared and evaluated as an efficient drug delivery system to improve the treatment of vaginal microbial infections. The liposome formulations were optimized based on sufficient trapping efficiencies for both drugs and membrane elasticity as a prerequisite for successful permeability and therapy. An appropriate viscosity for vaginal administration was achieved by incorporating the liposomes into Carbopol hydrogel. DPGLs were able to penetrate through the hydrogel network more rapidly than conventional liposomes. In vitro studies of drug release from the liposomal hydrogel under conditions simulating human treatment confirmed sustained and diffusion-based drug release. Characterization of the rheological and textural properties of the DPGL-containing liposomal hydrogels demonstrated that the incorporation of DPGLs alone had no significant influence on mechanical properties of hydrogels compared to controls. These results support the great potential of DPGL-in-hydrogel as an efficient delivery system for the controlled and sustained release of antimicrobial drugs in the vagina.     

A microprobe study of element distribution in vaginal epithelial cells of the rat

Microprobe analysis of vaginal epithelial cells shed during the estrous cycle of the rat was done to determine cellular elements present in successive stages: pro-estrus, estrus, and post-estrus. Smears of vaginal contents were placed on carbon planchettes, fixed by freeze-drying, and examined in a scanning microscope with an energy dispersive spectrometer. Concentrations of Na, Mg, P, S, Cl, K, and Ca were calculated (mmol/kg dry weight) and analyzed statistically. For phosphorus a significant fall at estrus correlates with a loss of nuclear and cytoplasmic nucleic acids and nucleoproteins. An increase in sulfur at estrus is consistent with an accumulation of keratins over pro-estrus and a greater increase over the post-estrus epithelial cells. The epithelial cells of pro-estrus are highest in Mg and Ca. By post-estrus, the cells have recovered their Mg, not Ca. Potassium concentrations exhibited no significant change between the successive stages.     

Discovery of Antimicrobial Peptides in Cervical‐Vaginal Fluid from Healthy Nonpregnant Women via an Integrated Proteome and Peptidome Analysis

Cervical‐vaginal fluid (CVF) covers the lower part of the female reproductive system and functions in the homeostasis and immunity of the surrounding tissues. In contrast to the CVF proteome of both nonpregnant and pregnant women, the CVF peptidome has not been reported to date. In the current study, we identified 1087 proteins in CVF, of which 801 proteins were not previously identified in CVF proteomes. The presence of the tissue‐specific proteins oviductal glycoprotein 1 and tubulin polymerization–promoting protein family member 3 strongly suggests that the tissues of the upper female reproductive tract contribute to the protein composition of CVF. The tremendous catalytic potential of CVF was highlighted by the identification of 85 proteases and the detection of pH‐dependent trypsin‐like proteolytic activity. Over 1000 endogenous peptides were detected in the CVF peptidome, and 39 peptides are predicted to have antimicrobial activity. The detailed proteomic and peptidomic analysis of CVF will further aid in the delineation of pathways related to reproduction, immunity and host defense, and assist in developing new biomarkers for malignant and other diseases of the female reproductive tract. Data are available via ProteomeXchange with identifiers PXD004450 (CVF peptidome) and PDX004363 (CVF proteome).     

A simple method for studying whole sections of rice grain

We developed a new and simple method to collect sections of a whole brown rice kernel for investigation of histological properties. A single kernel of rice was dehydrated through a graded ethanol series, transferred to xylene, and embedded in paraffin. During sectioning of the blocks using a rotary microtome, we used a special adhesive tape to collect and place the sections on slides so they remained flat. The use of the adhesive tape technique combined with autofluorescence characteristics allowed us to visualize cell walls throughout an entire longitudinal or transverse section of a whole rice kernel. We obtained scanning electron microscopy images of the sections to determine section quality.