Intravaginal treatment with Marantodes pumilum (Kacip Fatimah) ameliorates vaginal atrophy in rats with post-menopausal condition.

ETHNOPHARMACOLOGICAL RELEVANCE: Marantodes pumilum (MP) (Kacip Fatimah) is used to maintain the well-being of post-menopausal women. However, its role in ameliorating post menopause-related vaginal atrophy (VA) is unknown. AIMS: To investigate the ability of intravaginal MP gel treatment to ameliorate VA in sex-steroid deficient condition, mimicking post-menopause. METHODS: Ovariectomized female Sprague-Dawley rats received MP (100 µg/ml, 250 µg/ml and 500 µg/ml) and estriol (E) gels intravaginally for seven consecutive days. Rats were then euthanized and vagina was harvested and subjected for histological and protein expression and distribution analyses. Vaginal ultrastructure was observed by transmission electron microscopy (TEM). RESULTS: Thickness of vaginal epithelium increased with increasing intravaginal MP doses. Additionally, increased in expression and distribution of proliferative protein i.e. PCNA, tight junction protein i.e. occludin, water channel proteins i.e. AQP-1 and AQP-2 and proton extruder protein i.e. V-ATPase A1 were observed in the vagina following intravaginal MP and E gels treatment. Intravaginal MP and E gels also induced desmosome formation and approximation of the intercellular spaces between the vaginal epithelium. CONCLUSIONS: Intravaginal MP was able to ameliorate features associated with VA; thus, it has potential to be used as an agent to treat this condition.

1.7 micron optical coherence tomography for vaginal tissue characterization in vivo.

OBJECTIVES: Optical coherence tomography (OCT) can noninvasively visualize in vivo tissue microstructure with high spatial resolution that approaches the histologic level. Currently, OCT studies in gynecology are few and limited to a conventional 1.3 µm center wavelength swept light source which provides high spatial resolution but limited penetration depth. Here, we present a novel endoscopic OCT system with improved penetration depth and high resolution. METHODS: A novel endoscopic OCT system was developed based on a 1.7 µm swept source laser, which is capable of deeper tissue penetration due to its longer wavelength. To evaluate the performance of system, we imaged the human vaginas in vivo with both conventional 1.3 and 1.7 µm endoscopic OCT systems. RESULTS: With the 1.7 µm endoscopic OCT system, imaging depth was improved by more than 25%, allowing better visualization of the lamina propria and clear contrast of the epithelial layer from the surrounding tissues. CONCLUSION: The significantly improved performance of the novel 1.7 µm OCT imaging system demonstrates its potential use as a minimally-invasive monitoring tool of vaginal health in gynecologic practice. Lasers Surg. Med. 51:120-126, 2019. © 2018 Wiley Periodicals, Inc.

The first ultrastructural observations of the egg-forming complex of Calicotyle affinis, an endoparasitic monocotylid monogenean which lacks a uterus.

This ultrastructural study of the female reproductive system of Calicotyle affinis, a monogenean without a uterus, demonstrates, for the first time in a monogenean, the presence of accessory cells located in the peripheral region of the ovary and a single large cell whose surface is penetrated by deep invaginations which restrict the passage of oocytes through the lumen of the distal extremity of the ovary. The cytoarchitecture of the epithelial lining of the two vaginae is a syncytium formed by an anucleate epithelial lining with sunken epithelial perykaria. The wall of the seminal receptacle is an enlarged prolongation of the vaginal epithelial lining. Fertilization occurs in the fertilization chamber and fertilized oocytes retain cortical granules within their cytoplasm, a characteristic known to occur in free-living flatworms. Our study also highlights concentrations of two distinct groups of Mehlis' gland cell ducts on either side of the proximal end of the ootype distinguished by ultrastructural characteristics of their secretory granules. The epithelial wall of the ootype is formed by a single layer of regular, columnar, glandular epithelial cells; these cells are closely adjacent, conjoined towards their apical region by septate junctions and produce rounded, electron-dense granules which are discharged into the ootype lumen via a merocrine or holocrine mechanism. Released granules concentrate around the eggshell and form an additional fibrous coat. The morphological diversity observed in the female reproductive system of the Monogenea is commented on it relation to clarifying patterns in monogenean evolution and for understanding the phylogeny of the Neodermata.

Nystatin enhances the immune response against Candida albicans and protects the ultrastructure of the vaginal epithelium in a rat model of vulvovaginal candidiasis.

BACKGROUND: Vulvovaginal candidiasis (VVC) is a common infectious disease of the lower genital tract. Nystatin, a polyene fungicidal antibiotic, is used as a topical antifungal agent for VVC treatment. The aim of the current study was to investigate the possible immunomodulatory effects of nystatin on the vaginal mucosal immune response during Candida albicans infection and examine its role in protection of vaginal epithelial cell (VEC) ultrastructure. RESULTS: Following infection with C. albicans, IFN-γ and IL-17 levels in VECs were significantly elevated, while the presence of IgG was markedly decreased as compared to uninfected controls (P <  0.05). No significant differences in IL4 expression were observed. After treatment with nystatin, the level of IFN-γ, IL-17 and IgG was dramatically increased in comparison to the untreated group (P <  0.05). Transmission electron microscopy revealed that C. albicans invades the vaginal epithelium by both induced endocytosis and active penetration. Nystatin treatment protects the ultrastructure of the vaginal epithelium. Compared with the untreated C. albicans-infected group, Flameng scores which measure mitochondrial damage of VECs were markedly decreased (P <  0.001) and the number of adhesive and invasive C. albicans was significantly reduced (P <  0.01) after treatment with nystatin. CONCLUSIONS: Nystatin plays a protective role in the host defense against C. albicans by up-regulating the IFN-γ-related cellular response, the IL-17 signaling pathway and possibly through enhancing VEC-derived IgG-mediated immunity. Furthermore, nystatin notably improves the ultramorphology of the vaginal mucosa, partially through the protection of mitochondria ultrastructure in VECs and inhibition of adhesion and invasion by C. albicans. Together, these effects enhance the immune response of the vaginal mucosa against C. albicans and protect the ultrastructure of vaginal epithelium in VVC rats.

Merkel-like cell distribution in the epithelium of the human vagina. An immunohistochemical and TEM study.

Human Merkel cells (MCs) were first described by Friedrich S. Merkel in 1875 and named "Tastzellen" (touch cells). Merkel cells are primarily localized in the basal layer of the epidermis and concentrated in touch-sensitive areas. In our previous work, we reported on the distribution of MCs in the human esophagus, so therefore we chose other parts of the human body to study them. We selected the human vagina, because it has a similar epithelium as the esophagus and plays very important roles in reproduction and sexual pleasure. Due to the fact that there are very few research studies focusing on the innervation of this region, we decided to investigate the occurrence of MCs in the anterior wall of the vagina. The aim of our research was to identify MCs in the stratified squamous non-keratinized epithelium of the human vagina in 20 patients. For the identification of Merkel cells by light microscopy, we used antibodies against simple-epithelial cytokeratins (especially anti-cytokeratin 20). We also tried to identify them using transmission electron microscopy. Our investigation confirmed that 10 (50 %) of 20 patients had increased number of predominantly intraepithelial CK20 positive "Merkel-like" cells (MLCs) in the human vaginal epithelium. Subepithelial CK20 positive MLCs were observed in only one patient (5%). We tried to identify them also using transmission electron microscopy. Our investigation detected some unique cells that may be MCs. The purpose of vaginal innervation is still unclear. There are no data available concerning the distribution of MCs in the human vagina, so it would be interesting to study the role of MCs in the vaginal epithelium, in the context of innervation and epithelial biology.

Structural Reorganization of the Vaginal Mucosa in Stress Urinary Incontinence under Conditions of Er:YAG Laser Treatment.

Structural characteristics of the vaginal mucosa in stress incontinence and its correction by IncontiLase technology were studied. Studies of vaginal biopsy specimens before the exposure showed degenerative and atrophic changes in the stratified squamous epithelium, disorganization of fibrillar structures of the intercellular matrix, and microcirculatory disorders. Studies after Er:YAG laser exposure showed signs of neocollagenogenesis and elastogenesis, foci of neoangiogenesis, reduction of epithelial degeneration and atrophy, and an increase of the fibroblast population. Morphometry showed that the volume density of blood capillaries and the thickness of the epithelial layer increased by 61.1 and 64.5%, respectively. The use of IncontiLase technology in stress incontinence led to structural reorganization of the vaginal mucosa, improving its morphology and function and alleviating the symptoms of incontinence.

Cellular Evidence of Exosomes in the Reproductive Tract of Chinese Soft-Shelled Turtle Pelodiscus sinensis.

The oviduct is a location of egg production, fertilization, and sperm storage. While its secretions have broadly attributes toward different physiological functions. We examined the ultrastructure of oviduct epithelium and glands in relation to the secretions, particularly with exosomes origin in Chinese soft-shelled turtle Pelodiscus sinensis using immunohistochemistry and transmission electron microscopy. The ciliated epithelial and gland cells were involved in the release of exosomes and secretions into lumen throughout the year. The exosomes were either released directly from epithelium or in relation with multivesicular body (MVB). The average size of the particles varies between 50 and 130 nm. These exosomes were also widely distributed in the epithelial ciliated cells and pericytoplasm of glands lumen. Intracellular MVB was characterized by membrane-bounded exosomes of different sizes. Exosomes were also found in close contact with the cilia and sperm membrane in the lumen, which is suggestive of their fusogenic properties. Immunohistochemistry results showed strong to moderate positive expression of exosomes, in ciliated and gland cells, during January, September, and December, as it is the time of sperm storage in this turtle, whereas they showed moderate to weak level of expression during breeding season (May). This is first study about identification of the exosomes in female turtles. Epithelial and glandular exosomes, intracellular MVB, secretions, and secretory vesicles give this turtle specie a unique secretory morphology and a potential model for investigating the secretory nature of the oviduct.

Novel three-dimensional autologous tissue-engineered vaginal tissues using the self-assembly technique.

Many diseases necessitate the substitution of vaginal tissues. Current replacement therapies are associated with many complications. In this study, we aimed to create bioengineered neovaginas with the self-assembly technique using autologous vaginal epithelial (VE) and vaginal stromal (VS) cells without the use of exogenous materials and to document the survival and incorporation of these grafts into the tissues of nude female mice. Epithelial and stromal cells were isolated from vaginal biopsies. Stromal cells were driven to form collagen sheets, 3 of which were superimposed to form vaginal stromas. VE cells were seeded on top of these stromas and allowed to mature in an air-liquid interface. The vaginal equivalents were implanted subcutaneously in female nude mice, which were sacrificed after 1 and 2 weeks after surgery. The in vitro and animal-retrieved equivalents were assessed using histologic, functional, and mechanical evaluations. Vaginal equivalents could be handled easily. VE cells formed a well-differentiated epithelial layer with a continuous basement membrane. The equivalent matrix was composed of collagen I and III and elastin. The epithelium, basement membrane, and stroma were comparable to those of native vaginal tissues. The implanted equivalents formed mature vaginal epithelium and matrix that were integrated into the mice tissues. Using the self-assembly technique, in vitro vaginal tissues were created with many functional and biological similarities to native vagina without any foreign material. They formed functional vaginal tissues after in vivo animal implantation. It is appropriate for vaginal substitution and disease modeling for infectious studies, vaginal applicants, and drug testing.

Characterization of the ultrastructure in the uterovaginal junction of the hen.

In poultry, the infundibulum is the place of fertilization, eggshell production, and sperm storage, while its uterovaginal junction (UVJ) is regarded as the most important site, which has abundant sperm storage tubules (SST). We examined the ultrastructure of the epithelium with relation to its unique secretory cytology in the UVJ of hens using transmission electron microscopy. The epithelium of the UVJ is lined with ciliated and secretory cells. Ciliated cells are characterized with light and dense secretory granules in supernuclear cytoplasm. Dense secretory granules in ciliated cells are larger in diameter (one µm), surrounded with a transparent rim and concentric layers, whereas the dense granules in the ciliated cells of SST are smaller (0.52 µm) in size and not surrounded by any transparent rim or layer. Ciliated cells also are involved in the shedding of exosomes and secretory vesicles in the lumen. Secretory exosomes are in close contact with cilia and directly release from the apical border into the lumen. Cell junctions are widely distributed between these cells. The secretory cells are associated with the release of secretions via apocrine blebs from microvilli and secretory vesicles, which protrude out from the surface of the epithelium. The dense secretory granules in these cells are smaller in size (0.6 µm), absent of a transparent rim or layers, and are released into the lumen through secretory vesicles. The intracellular multivesicular body (MVB) also is observed in the supernuclear cytoplasm of secretory cells, which are related to the production of exosomes. In general, the apical protrusion of the epithelium in the form of apocrine secretions, the releasing of exosomes, the identification of intracellular MVB, and the release of dense granules give the epithelium a distinct morphology in the UVJ of the hen oviduct.

Propylene glycol-embodying deformable liposomes as a novel drug delivery carrier for vaginal fibrauretine delivery applications.

The purpose of this work was to develop and characterize the fibrauretine (FN) loaded propylene glycol-embodying deformable liposomes (FDL), and evaluate the pharmacokinetic behavior and safety of FDL for vaginal drug delivery applications. FDL was characterized for structure, particle size, zeta potential, deformability and encapsulation efficiency; the ability of FDL to deliver FN across vagina tissue in vitro and the distribution behavior of FN in rat by vaginal drug delivery were investigated, the safety of FDL to the vagina of rabbits and rats as well as human vaginal epithelial cells (VK2/E6E7) were also evaluated. Results revealed that: (i) the FDL have a closed spherical shape and lamellar structure with a homogeneous size of 185±19nm, and exhibited a negative charge of -53±2.7mV, FDL also have a good flexibility with a deformability of 92±5.6 (%phospholipids/min); (ii) the dissolving capacity of inner water phase and hydrophilicity of phospholipid bilayers of deformable liposomes were increased by the presence of propylene glycol, this may be elucidated by the fluorescent probes both lipophilic Nile red and hydrophilic calcein that were filled up the entire volume of the FDL uniformly, so the FDL with a high entrapment capacity (were calculated as percentages of total drug) for FN was 78±2.14%; (iii) the permeability of FN through vaginal mucosa was obviously improved by propylene glycol-embodying deformable liposomes, no matter whether the FN loaded in liposomes or not, although FN loaded in liposomes caused the highest permeability and drug reservoir in vagina; (iv) the FN mainly aggregated in the vagina and uterus, then the blood, spleen, liver, kidney, heart and lungs for vaginal drug delivery, this indicating vaginal delivery of FDL have a better 'vaginal local targeting effect'; and (v) the results of safety evaluation illustrate that the FDL is non-irritant and well tolerated in vivo, thereby establishing its vaginal drug delivery potential. These results indicate that the propylene glycol-embodying deformable liposomes may be a promising drug delivery carrier for vaginal delivery of fibrauretine.

Novel cellular evidence of oviduct secretions in the Chinese soft-shelled turtle Pelodiscus sinensis.

The oviduct is the location of fertilization and sperm storage. We examined the ultrastructure of the oviduct epithelium and its glandular secretions in the isthmus, uterus and vagina of Chinese soft-shelled turtle Pelodiscus sinensis using light and transmission electron microscopy. The epithelium in these segments is lined with ciliated, secretory and other cells; the first two cell types span the entire epithelium, with secretory cells being predominant. The ciliated cells are characterized by the presence of a secretory vacuole that releases apocrine secretions into the lumen, whereas the secretory cells contain typical biphasic granules with both dark and light aspects. The third type of cells observed have wider proximal portion, abundant mitochondria, vacuoles, and narrow nuclei. The storage of spermatozoa is restricted to the isthmus, uterus, and vagina. In addition, the gland cells show prominent features, including the presence of granules of different shapes, sizes, and electron densities. The synthesis of these granules is described for the first time in this study. Mitochondria appear to play an important role in the formation of dense granules, the rough endoplasmic reticulum and microfilaments may also play a role in the maturation of these dense granules. After completing the maturation process, these granules are released into the lumen of the gland cells.

Apical blebs on sperm storage tubule epithelial cell microvilli: their release and interaction with resident sperm in the turkey hen oviduct.

Located at the anterior end of the turkey hen's vagina are numerous discrete tubular invaginations of the surface epithelium, collectively referred to as the sperm storage tubules (SSTs). After mating or artificial insemination, sperm ascend the vagina, enter the SSTs, and over the ensuing days and weeks, gradually exit the SSTs and are transported to the anterior end of the oviduct to fertilize a daily succession of ova. Little is known regarding the cellular and molecular mechanisms responsible for sperm subsistence in the lumen of the SST. In this study, the origin of microvillus blebs (MvBs) on the apical tips of SST epithelial cells was examined, and their possible role in sperm survival was discussed. Regardless, if sperm are present or not, transmission electron microscopy revealed two types of microvilli differentiated by the presence or absence of pleomorphic unilaminar MvBs localized to their apical tips. Although some MvBs appeared to be discharging their contents into the SST lumen, others appeared to have pinched off the microvillus stem. When SSTs contained clusters of densely packed sperm, the sperm heads of those sperm adjacent to the SST epithelial cell surface were surrounded by the microvilli. Associated with the plasmalemma of sperm throughout the SST lumina were membrane fragments and small vesicles (30-130 nm in diameter), some of which appeared to have fused with sperm. It is concluded that the MvBs are a form of shedding vesicle released from the SST epithelial cell microvilli by apocrine secretion. On the basis of observations described herein and those of other authors, it is suggested that the MvBs contribute to sustained sperm storage in the SSTs by (1) supplying metabolic substrates used by resident sperm, (2) serving as fusogenic vehicles providing exogenous macromolecules that reversibly suppress sperm functions associated with fertilization (decapacitation?) and stabilize the sperm plasmalemma, and (3) acting as transport vesicles actively transporting fluid from the SST epithelial cells to the SST lumen.

A new species of diplectanid (Monogenoidea) from Paranthias colonus (Perciformes, Serranidae) off Peru.

Pseudorhabdosynochus jeanloui n. sp. (Monogenoidea, Diplectanidae) is described from specimens collected from the gills of the Pacific creolefish, Paranthias colonus (Perciformes, Serranidae) from a fish market in Chorrillos, Lima, Peru. The new species is differentiated from other members of the genus by the structure of its sclerotized vagina, which has two spherical chambers of similar diameter. This is the first Pseudorhabdosynochus species described from the Pacific coast of America, the third species of the genus reported from South America and the first described from a member of Paranthias.

Pseudorhabdosynochus regius n. sp. (Monogenea, Diplectanidae) from the mottled grouper Mycteroperca rubra (Teleostei) in the Mediterranean Sea and Eastern Atlantic.

Pseudorhabdosynochus regius n. sp. is described from the gills of the mottled grouper Mycteroperca rubra caught off Senegal, Tunisia and Libya (type-locality: off Dakar, Senegal). The species is distinguished from its congeners by the structure of its sclerotised vagina (length 26-35 µm), which exhibits a trumpet in continuity with the primary canal, a straight primary canal, and primary and secondary chambers included in a common sclerotised mass along the primary canal. The species is also characterised by small squamodiscs (length 20-40 µm) with 10-11 rows of rodlets. Its closest relatives (based on the structure of the sclerotised vagina) are species mostly found in the Mediterranean Sea and parasites on species of Mycteroperca. A second species of Pseudorhabdosynochus Yamaguti, 1958 is reported from the same host and localities but not described. A list of diplectanids from groupers in the Mediterranean Sea is provided. We point out that a recent article was not compliant with the new Article 8.5.3 of the International Code of Zoological Nomenclature; for this reason, three species, P. nhatrangensis Dang, Bristow, Schander & Berland, 2013, P. vietnamensis Dang et al., 2013 and P. brunei Dang et al., 2013, are invalid.

Microscopic and ultrastructural modifications of postmenopausal atrophic vaginal mucosa after fractional carbon dioxide laser treatment.

Vaginal atrophy occurring during menopause is closely related to the dramatic decrease in ovarian estrogens due to the loss of follicular activity. Particularly, significant changes occur in the structure of the vaginal mucosa, with consequent impairment of many physiological functions. In this study, carried out on bioptic vaginal mucosa samples from postmenopausal, nonestrogenized women, we present microscopic and ultrastructural modifications of vaginal mucosa following fractional carbon dioxide (CO2) laser treatment. We observed the restoration of the vaginal thick squamous stratified epithelium with a significant storage of glycogen in the epithelial cells and a high degree of glycogen-rich shedding cells at the epithelial surface. Moreover, in the connective tissue constituting the lamina propria, active fibroblasts synthesized new components of the extracellular matrix including collagen and ground substance (extrafibrillar matrix) molecules. Differently from atrophic mucosa, newly-formed papillae of connective tissue indented in the epithelium and typical blood capillaries penetrating inside the papillae, were also observed. Our morphological findings support the effectiveness of fractional CO2 laser application for the restoration of vaginal mucosa structure and related physiological trophism. These findings clearly coupled with striking clinical relief from symptoms suffered by the patients before treatment.

Morphological studies on the vagina and vestibule of dromedary camels (Camelus dromedarius) / Estudios morfológicos de la vagina y vestíbulo del camello (Camelus dromedarius)

The camel (Camelus dromedarius) is an important multipurpose livestock species and its meat represents about 10% of the red meat consumption in Egypt. The reproductive efficiency of camel under natural condition is generally considered to be low. Sound knowledge about the tubular genital organs of this species might facilitate the application of new reproductive methodology. Our study was therefore conducted to investigate the morphology of mucosal surface of vagina and vestibule on camels using light, scanning and transmission electron microscopy. The mucosal surface of vagina consisted of stratified columnar epithelium with mucous secreting cells (goblet-like cells). SEM of vagina revealed the presence of longitudinal primary and secondary folds and small transverse folds. The columnar epithelium showed marked cell boundary and its apical surface was studded by a lot of microvilli. TEM confirmed the presence of microvilli at apical surfaces and showed some secretory granules in the supranuclear region of the columnar epithelium. The vestibule of dromedary camel was lined by stratified squamous keratinized epithelium. Basal lamina and stratum granulosum of this epithelium showed strong PAS positive reaction. SEM of vestibule revealed the presence of small longitudinal and fine transverse folds with a lot of mucous debris. However TEM of vestibule showed the stratified squamous keratinized epithelium with basal layer of cuboidal cells and superficial layers of squamous cells.

El camello (Camelus dromedarius) es una importante especie de ganado de usos múltiples y el consumo de su carne corresponde al 10% aproximadamente del consumo de carne roja en Egipto. La eficiencia reproductiva del camello, bajo condiciones naturales, se considera generalmente baja. El conocimiento adecuado sobre los órganos genitales tubulares de esta especie podría facilitar la aplicación de una nueva metodología de reproducción. Por lo tanto, se llevó a cabo este estudio para investigar la morfología de la superficie de la mucosa de la vagina y el vestíbulo en camellos, utilizando luz, escaneado y microscopía electrónica de transmisión. La superficie de la mucosa de la vagina está formado por epitelio columnar estratificado con células secretoras mucosas (células en copa). La microscopía electrónica de barrido (SEM) de la vagina reveló la presencia de pliegues primarios y secundarios longitudinales y pequeños pliegues transversales. El epitelio columnar mostró un límite celular marcado y su superficie apical se evidenció salpicado por una gran cantidad de microvellosidades. La microscopía electrónica por transmisión (TEM) confirmó la presencia de microvellosidades en las superficies apicales y mostró algunos gránulos secretores en la región supranuclear del epitelio columnar. El vestíbulo del dromedario está revestido por epitelio estratificado queratinizado, de tipo escamoso. La lámina basal y el estrato granuloso de este epitelio mostraron una fuerte reacción PAS positiva. La microscopía electrónica de barrido (SEM) del vestíbulo reveló la presencia de pequeños pliegues transversales longitudinales y finos, con gran cantidad de restos de mucosidad. Sin embargo, la microscopía electrónica por transmisión (TEM) del vestíbuloreveló un epitelio queratinizado escamoso estratificado, con una capa basal de células cúbicas y capas superficiales de células escamosas.

Ultrastructural features of vagina at different phases of the oestrous cycle in the female African giant rat (Cricetomys gambianus Waterhouse).

The ultrastructures of the vagina at various stages of the oestrous cycle in female African giant rats (Cricetomys gambianus Waterhouse) were described in the present study. At mid-proestrus, late proestrus (LP)/early estrus (EE) and mid-estrus (ME) as well as late metestrus (LM)/early diestrus (ED) and mid-diestrus (MD), complex interface of epithelium and lamina propria were observed. Cells of the stratum basale formed finger-like extensions into the underlying lamina propria and tips of the extensions displayed hemidesmosome while basal lamina followed the contour of the extensions. At mid-metestrus (MM) and late diestrus/early proestrus, well developed, relatively straight basal lamina interfaced between the stratum basale and the lamina propria without finger-like projections. Polygonal cells with indented nuclei and, cytoplasm containing ribosomes, polysomes, intermediate filaments, and mitochondria were observed in stratum spinosum at all the phases of the oestrus cycle. At MM, LM/ED, and MD, the stratum spinosum had numerous desmosomes with tonofilaments, large microvilli that intermingled at the intercellular spaces and evidence of trapped/migrating neutrophils and lymphocytes. The superficial layer displayed short microvilli at mid-proestrus, cornification at LP/EE and desquamation at ME while it showed condensation of intermediate filaments; projections of large microvilli into the luminal surface at MM, and embeddement of neutrophils at LM/ED as well as MD. This study looked into the reproductive biology of female African giant rats to produce baseline information on its reproductive organs and represented the first comprehensive description of the vagina at the ultrastructural level during oestrous cycle.

The case of a prosthetic limb used to cause lethal intravaginal injuries: forensic medical aspects in a case of intimate partner violence.

A common form of violence against women is sexual coercion on the part of their husbands/partners, the uncontrollable effects of which can lead to extreme consequences, as in the case of uxoricide examined in this report. It involved a 59-year-old female alcoholic, under observation on the part of social services as the possible victim of abuse by her husband, an amputee with a transtibial prosthesis. The woman had never admitted to her social workers that her husband was abusing her. One night, she was admitted to hospital in a state of hemorrhagic shock due to massive vaginal bleeding, but despite treatment, she died 20 min after arrival. The anatomical-pathological examination conducted by the hospital revealed serious genital lesions which warranted reporting the case to the Judicial Authorities, who arranged for a forensic autopsy. The cause of death was identified as acute meta-hemorrhagic anemia in a cirrhotic woman, secondary to a large, irregular vaginal lesion involving both the vaginal wall and the soft perivaginal tissues as well as the medium and small urogenital vascular branches. To identify the foreign body used to inflict this injury, a scanning electron microscope and energy dispersive X-ray spectrometer (SEM-EDS) were used. This revealed tiny splinters of wood in the vaginal tissues examined. In addition to the genetic-forensic techniques used, this finding allowed the investigators to identify the husband's prosthetic limb as the instrument of sexual coercion. The report describes a particular case of marital rape that resulted in uxoricide, in which the overall concordance of the investigations carried out played a fundamental role in identifying the offending body and, consequently, the murderer.

Expression and secretion of plasma membrane Ca2+-ATPase 4a (PMCA4a) during murine estrus: association with oviductal exosomes and uptake in sperm.

PMCA4, a membrane protein, is the major Ca(2+) efflux pump in murine sperm where its deletion leads to a severe loss of hyperactivated motility and to male infertility. We have previously shown that the PMCA4b splice variant interacts with CASK (Ca(2+/)CaM-dependent serine kinase) in regulating sperm Ca(2+). More recently we detected that PMCA4a isoform, in addition to its presence in testis, is secreted in the epididymal luminal fluid and transferred to sperm. Here we show that Pmca4 mRNA is expressed in both the 4a and 4b variants in the vagina, uterus, and oviduct. Immunofluorescence reveals that PMCA4a is similarly expressed and is elevated during estrus, appearing in the glandular and luminal epithelia. Western analysis detected PMCA4a in all tissues and in the luminal fluids (LF) of the vagina (VLF), uterus (ULF), and the oviduct (OLF) collected during estrus. It was ~9- and 4-fold higher in OLF than in VLF and ULF, and only marginally present in LF collected at metestrus/diestrus. Fractionation of the LF collected at estrus, via ultracentrifugation, revealed that 100% of the PMCA4a resides in the vesicular fraction of the ULF and OLF. Transmission electron microscopy (TEM) revealed that OLF vesicles have an exosomal orientation (with the cytoplasmic-side inward), a size range of 25-100 nm, with the characteristic CD9 biomarker. Thus, we dubbed these vesicles "oviductosomes", to which PMCA4a was immunolocalized. Incubation of caudal sperm in the combined LF or exosomes resulted in up to a ~3-fold increase of sperm PMCA4a, as detected by flow cytometry, indicating in vitro uptake. Our results are consistent with the increased requirement of Ca(2+) efflux in the oviduct. They show for the first time the presence of oviductal exosomes and highlight their role, along with uterosomes and vaginal exosomes, in post-testicular sperm acquisition of PMCA4a which is essential for hyperactivated motility and fertility.

Ultrastructural characteristics of the vaginae of the basal monogenean Chimaericola leptogaster (Leuckart, 1830).

In the first study of the vaginal ultrastructure of any monogenean, the paired vaginae of Chimaericola leptogaster, a basal polyopisthocotylean from the gills of a holocephalan, are described. Each vagina opens laterally. A unique feature of this relict parasite is a short vagino-intestinal connecting duct, the lining of which is separated by septate junctions from the linings of the vagina and the intestine. After giving rise to this vagino-intestinal duct, each vagina travels in a posterior direction and opens into the vitelline collecting duct. The lining of each vagina close to the vaginal pore resembles the body tegument, the syncytial lining of which measures about 2.5 µm in thickness and has a dense surface layer 0.2 µm in thickness and different apical projections. Proximal to this and before the entrance to the vaginal-intestinal duct, the vaginal lining is characterised by: uniformly distributed electron-dense, lanceolate, spine-like surface structures of about 1.7 µm in length; three kinds of surrounding glands with three kinds of secretory granules (abundant oval or rounded electron-dense granules 0.7-2.5 µm in length, much less numerous spheroidal, vesicular granules 0.3-0.4 µm in diameter, and tightly packed, spheroidal granules 0.8 µm in diameter and containing fine particulate material of low density); and four kinds of sensory receptors (three uniciliate and one aciliate). These features are absent from the rest of the vagina. The likely roles of the vagino-intestinal connection, the different gland cells, sensory structures, and the armament of the distal vaginal regions are discussed. Considering the suggested polyopisthocotylean ancestor for the Neodermata, the relict parasite fauna of holocephalans and the unique vagino-intestinal connection in C. leptogaster, which is also known in turbellarians, an ancient origin for chimaericolids is supported.