Sperm Localization in the Oviducts of Artificially Inseminated Dairy Cattle

Eight animals, 3 heifers and 5 primiparous cows, were artificially inseminated by intrauterine deposition of frozen-thawed semen. The insemination dose comprised 20×106 or 200 × 106 spermatozoa, frozen in French mini straws. Four animals were inseminated at fixed time interval (72 or 84 h) after cloprostenol injection. The remaining 4 animals were inseminated in spontaneous oestrus. Slaughter took place 2 or 12 h after insemination. After fixation the oviducts were cut into segments, which were serial-sectioned and stained. Six sections per segment were examined under the microscope for sperm recovery. The number of spermatozoa recovered from the oviducts varied considerably among animals. Recovery was poor (less than 50 spermatozoa) in 4 animals. Recovery was low when insemination took place in induced oestrus and with the lower sperm number (20×106). In animals in which more than 50 spermatozoa were found the distribution varied both between animals and between oviducts within the same animal. Overall, more spermatozoa were found in the lower (UTJ, isthmus and AIJ) than in the upper (ampulla) parts of the oviducts. In 3 out of 4 animals more spermatozoa were recovered from the left than from the right oviduct. Only in 1 animal were the majority of spermatozoa found in the oviduct ipsilateral to the follicle-bearing ovary.     

R5 Macrophage-Tropic HIV-1 in the Male Genital Tract

The entry tropism of HIV-1 Env proteins from virus isolated from the blood and genital tract of five men with compartmentalized lineages was determined. The Env proteins isolated from the genital tract of subject C018 were macrophage-tropic proteins, while the remaining cloned env genes encoded R5 T cell-tropic proteins. The detection of a macrophage-tropic lineage of HIV-1 within the male genital tract strongly suggests that evolution of macrophage-tropic viruses can occur in anatomically isolated sites outside the central nervous system.     

女性生殖道炎症支原体多重耐药性分析

了解本地区在采用常规抗生素治疗支原体后多重耐药株的产生情况,为临床针对性用药提供科学依据。采用培养法对女性生殖道炎患者分泌物进行支原体培养、体外药敏试验及分析。结果可见,感染数量≥10~4 cfu/mL的支原体阳性率为52.9%(82/155);13种抗生素中,耐药种数构成比为6R>2R、4R、5R>3R>1R>8R>7R、10R>0R>9R、12R、13R;耐药率为林可霉素>壮观霉素>环丙沙星>诺氟沙星>司帕沙星>罗红霉素>氧氟沙星>克拉霉素>左氧氟沙星>阿齐霉素>强力霉素,美满霉素>交沙霉素。体外药敏试验对临床治疗支原体感染及减少多重耐药株的产生具有重要指导意义。     

p16 Expression in Squamous Lesions of the Female Genital Tract

The aim of this study was to examine the role of p16 in the pathogenesis of squamous carcinoma of the gynecologic tract. Squamous carcinoma and carcinoma in situ from the female genital tract were examined for the expression of p16 by paraffin immunohistochemistry. About 74% (40/54) of cases showed p16 expression. By primary site, 77% (23/30) of cervical, 67% (6/9) of vaginal and 85% (11/13) of vulvar primaries expressed p16, but two endometrial primary squamous carcinomas were negative (0/2). In addition, p16 was not identified in non-dysplastic tissue and low grade dysplasia. In cases where there were matched vaginal or vulvar and cervical primaries in a given patient, there was concordant positive p16 expression. It is concluded that p16 is frequently expressed in squamous carcinoma of the cervix, vagina and vulva, but not seen in cases of benign and low grade lesions. It may be a marker of transformation from a low to a high grade lesion. More cases of endometrial primaries need to be studied to see if these evolve by a p16-independent pathway.     

Isolation of Lymphocytes from Mouse Genital Tract Mucosa

Mucosal surfaces, including in the gastrointestinal, urogenital, and respiratory tracts, provide portals of entry for pathogens, such as viruses and bacteria ¹. Mucosae are also inductive sites in the host to generate immunity against pathogens, such as the Peyers patches in the intestinal tract and the nasal-associated lymphoreticular tissue in the respiratory tract. This unique feature brings mucosal immunity as a crucial player of the host defense system. Many studies have been focused on gastrointestinal and respiratory mucosal sites. However, there has been little investigation of reproductive mucosal sites. The genital tract mucosa is the primary infection site for sexually transmitted diseases (STD), including bacterial and viral infections. STDs are one of the most critical health challenges facing the world today. Centers for Disease Control and Prevention estimates that there are 19 million new infectious every year in the United States. STDs cost the U.S. health care system $17 billion every year ², and cost individuals even more in immediate and life-long health consequences. In order to confront this challenge, a greater understanding of reproductive mucosal immunity is needed and isolating lymphocytes is an essential component of these studies. Here, we present a method to reproducibly isolate lymphocytes from murine female genital tracts for immunological studies that can be modified for adaption to other species. The method described below is based on one mouse.      

女性激素状态与生殖道支原体寄居的关系

本文调查了959名不同性激素状态妇女生殖道溶脲支原体(Uu)和人型支原体(Mh)的寄居情况。结果表明新生女婴、产后妇女和绝经妇女Uu和Mh寄居率较低,妊娠妇女Uu和Mh寄居率则相当高;已婚非孕妇女Uu和Mh寄居率高于未婚者。提示了女性激素水平与生殖道支原体寄居状态有密切联系。     

New Candidate Biomarkers in the Female Genital Tract to Evaluate Microbicide Toxicity

Vaginal microbicides hold great promise for the prevention of viral diseases like HIV, but the failure of several microbicide candidates in clinical trials has raised important questions regarding the parameters to be evaluated to determine in vivo efficacy in humans. Clinical trials of the candidate microbicides nonoxynol-9 (N9) and cellulose sulfate revealed an increase in HIV infection, vaginal inflammation, and recruitment of HIV susceptible lymphocytes, highlighting the need to identify biomarkers that can accurately predict microbicide toxicity early in preclinical development and in human trials. We used quantitative proteomics and RT-PCR approaches in mice and rabbits to identify protein changes in vaginal fluid and tissue in response to treatment with N9 or benzalkonium chloride (BZK). We compared changes generated with N9 and BZK treatment to the changes generated in response to tenofovir gel, a candidate microbicide that holds promise as a safe and effective microbicide. Both compounds down regulated mucin 5 subtype B, and peptidoglycan recognition protein 1 in vaginal tissue; however, mucosal brush samples also showed upregulation of plasma proteins fibrinogen, plasminogen, apolipoprotein A-1, and apolipoprotein C-1, which may be a response to the erosive nature of N9 and BZK. Additional proteins down-regulated in vaginal tissue by N9 or BZK treatment include CD166 antigen, olfactomedin-4, and anterior gradient protein 2 homolog. We also observed increases in the expression of C-C chemokines CCL3, CCL5, and CCL7 in response to treatment. There was concordance in expression level changes for several of these proteins using both the mouse and rabbit models. Using a human vaginal epithelial cell line, the expression of mucin 5 subtype B and olfactomedin-4 were down-regulated in response to N9, suggesting these markers could apply to humans. These data identifies new proteins that after further validation could become part of a panel of biomarkers to effectively evaluate microbicide toxicity.     

Microbiome Composition and Function Drives Wound-Healing Impairment in the Female Genital Tract

The mechanism(s) by which bacterial communities impact susceptibility to infectious diseases, such as HIV, and maintain female genital tract (FGT) health are poorly understood. Evaluation of FGT bacteria has predominantly been limited to studies of species abundance, but not bacterial function. We therefore sought to examine the relationship of bacterial community composition and function with mucosal epithelial barrier health in the context of bacterial vaginosis (BV) using metaproteomic, metagenomic, and in vitro approaches. We found highly diverse bacterial communities dominated by Gardnerella vaginalis associated with host epithelial barrier disruption and enhanced immune activation, and low diversity communities dominated by Lactobacillus species that associated with lower Nugent scores, reduced pH, and expression of host mucosal proteins important for maintaining epithelial integrity. Importantly, proteomic signatures of disrupted epithelial integrity associated with G. vaginalis-dominated communities in the absence of clinical BV diagnosis. Because traditional clinical assessments did not capture this, it likely represents a larger underrepresented phenomenon in populations with high prevalence of G. vaginalis. We finally demonstrated that soluble products derived from G. vaginalis inhibited wound healing, while those derived from L. iners did not, providing insight into functional mechanisms by which FGT bacterial communities affect epithelial barrier integrity.     

Mycoplasmosis: Experimental and Spontaneous Infections of the Genital Tract of Bulls

A strain of M. bovigenitalium was isolated from semen of a bull (K) with chronic seminal vesiculitis. Using this strain a vesiculitis of the same type as found in bull K, characterized by simultaneous acute and chronic lesions, was induced experimentally in 2 bulls by direct inoculation into the vesicular glands. In the acute phase a marked infiltration of eosinophils was found in the interstitial tissues and alveoli whereas in the chronic phase fibrosis, lymphoid and epithelial hyperplasia were seen. Degeneration of vascular walls and connective tissue was common. On inoculation into the testis of 3 bulls a chronic epididymitis and ampullitis were produced. The histological changes were of the same type as found in the vesicular glands of the 3 bulls with vesiculitis. By indirect hemagglutination a specific and significant increase in serum antibody could be demonstrated. As the titers were low and the maximum titers were reached early, it will probably not often be possible to make an etiological diagnosis on the basis of serological evidence. A comparative experiment employing the type strain (PG 11) of M. bovigenitalium was performed. A rise in antibody titer was seen, but neither clinical nor histological changes could be demonstrated.     

Ultrastructure of the Z-organ and Parts of the Female Genital Tract in Xiphinema coxi coxi

Ultrastructure of the Z-organ and associated apophyses in Xiphinema coxi coxi was studied by transmission electron microscopy to determine their structural origin and relationship with other parts of the genital tract. The Z-organ of X. coxi coxi is oval-shaped, ca. 30 µm long and 16 µm wide. It is clearly distinguished from the other parts of the female genital tract by its thick muscular outer wall, epithelium-lined lumen, and 4-5 centrally located apophyses. Each apophysis is continuous with the epithelial lining of the Z-organ, suggesting that it originated from epithelium. The apophyses appear as thickened and densely folded masses forming numerous interlaced pores and (or) chambers containing mucous-like materials and electron-dense crystals. These apophyses are characteristic of a typical Z-organ; no globular structures characteristics of the pseudo-Z-organ were observed. The thickness of the muscular layer of the oviduct and uterus varied with position. The overall Z-organ ultrastructure of this study, including body wall and internal apophyses, was comparable to the typical Z-organ of X. ifacolum. This suggests that X. coxi coxi should be classified as a Xiphinema species that contains the typical Z-organ.     

Viruses in the Mammalian Male Genital Tract and Their Effects on the Reproductive System

This review describes the various viruses identified in the semen and reproductive tracts of mammals (including humans), their distribution in tissues and fluids, their possible cell targets, and the functional consequences of their infectivity on the reproductive and endocrine systems. The consequences of these viral infections on the reproductive tract and semen can be extremely serious in terms of organ integrity, development of pathological and cancerous processes, and transmission of diseases. Furthermore, of essential importance is the fact that viral infection of the testicular cells may result not only in changes in testicular function, a serious risk for the fertility and general health of the individual (such as a fall in testosteronemia leading to cachexia), but also in the possible transmission of virus-induced mutations to subsequent generations. In addition to providing an exhaustive account of the data available in these domains, this review focuses attention on the fact that the interface between endocrinology and virology has so far been poorly explored, particularly when major health, social and economical problems are posed. Our conclusions highlight the research strategies that need to be developed. Progress in all these domains is essential for the development of new treatment strategies to eradicate viruses and to correct the virus-induced dysfunction of the endocrine system.     

Folate Receptors in Malignant and Benign Tissues of Human Female Genital Tract

We have characterized the folate receptor in malignant and benign tissues of human female genital tract (Fallopian tube and benign and malignant tissues of uterus). Radioligand binding displayed characteristics similar to those of other folate binding proteins. Those include a high-affinity type of binding (K=10¹⁰M^–1), apparent positive cooperativity, a slow dissociation at pH 7.4 becoming rapid at pH 3.5, and inhibition of binding by folate analogues. The gel filtration profile of Triton X-100 solubilized tissue contained two large peaks of ³H-folate labelled protein (>=130 and 100kDa) as well as a 25 kDa peak. Only a single band of 70 kDa was seen on SDS-PAGE immunoblotting. The large molecular size forms on gel filtration appear to represent folate receptors having a hydrophobic membrane anchor inserted into Triton X-100 micelles. The folate receptor of female genital tract showed cross-reactivity in ELISA and positive immunostaining with rabbit antibodies against human milk folate binding protein. Variations in the ratio of immunoresponse to total high affinity folic acid binding suggests the presence of multiple isoforms of the receptor in different types of malignant and benign tissues.