Interactions Between Spermatozoa and the Crypts, Cilia, and Mucus of the Cervix in the Ewe

In ruminants, minor cervical folds, commonly called crypts, temporarily store spermatozoa for a short interval of time between insemination and fertilization. However, the mechanism by which spermatozoa are transported to these crypts and subsequently to the uterus is not known. To study this problem, cervical tissue, which was removed from ewes that were naturally inseminated by rams at estrus, was examined with the scanning electron microscope to determine the physical associations that occur between the spermatozoa and the structural features of the cervix. The study indicates that the spermatozoa generally are not oriented parallel to the longitudinal axis of the cervix, exhibit no consistent association with the cervical cilia, and do not lie in any well defined channels formed by the cervical secretions. Alternatively, the majority of spermatozoa occur as isolated aggregations that lie in or near the shallow folds or crypts of the cervix. The vast numbers of spermatozoa in these aggregations and the lack of any common orientation suggest that some form of external stimulus, such as cervical contractions, might be responsible for the initial mass movement and distribution of spermatozoa in the cervix of the ewe

Increasing Resolution and Versatility in Low Temperature Conventional and Field Emission Scanning Electron Microscopy

Studies were undertaken to expand the versatility and the resolution of low temperature conventional and field emission scanning electron microscopy (SEM). The results indicated that simple modified specimen holders, which could be used in conjunction with the commercial cryosystems, allowed one to store specimens for several weeks in liquid nitrogen, either before or after observation in a conventional SEM, without incurring degradation of the surface features. Other modified holders permitted one to move the specimen closer to the final lens or to use the upper secondary electron detector, which is available with some SEMs. Both of these procedures increased the resolution that was attainable with the standard holders. In conventional SEM (CSEM) and field emission SEM (FESEM), holders were also modified to allow one to obtain complementary images of fractured specimens. When a conventional vacuum evaporator equipped with a freeze-etch module was used in conjunction with these holders, specimens could be fractured, etched, shadowed with platinum and coated with carbon before the sample was transferred to the cryostage in the SEM. This procedure increased resolution beyond that obtained with the sputter units in two commercial cryosystems that were used on a CSEM and a FESEM, provided membrane particle resolution in the FESEM and produced a coating or replica that could be recovered and examined in a TEM. These results, which demonstrated how resolution of cryospecimens can be enhanced in CSEM and FESEM, indicated that coating specimens in a high vacuum evaporator provided an alternative procedure that could be used to obtain high resolution images in a FESEM

Oncostatin M promotes STAT3 activation, VEGF production, and invasion in osteosarcoma cell lines

<p>Abstract</p> <p>Background</p> <p>We have previously demonstrated that both canine and human OSA cell lines, as well as 8 fresh canine OSA tumor samples, exhibit constitutive phosphorylation of STAT3, and that this correlates with enhanced expression of matrix metalloproteinase-2 (MMP2). While multiple signal transduction pathways can result in phosphorylation of STAT3, stimulation of the cytokine receptor gp130 through either IL-6 or Oncostatin M (OSM) is the most common mechanism through which STAT3 is activated. The purpose of this study was to evaluate the role of IL-6 and OSM stimulation on both canine and human OSA cell lines to begin to determine the role of these cytokines in the biology of OSA.</p> <p>Methods</p> <p>RT-PCR and Western blotting were used to interrogate the consequences of OSM and IL-6 stimulation of OSA cell lines. OSA cells were stimulated with OSM and/or hepatocyte growth factor (HGF) and the effects on MMP2 activity (gel zymography), proliferation (CyQUANT), invasion (Matrigel transwell assay), and VEGF production (Western blotting, ELISA) were assessed. The small molecule STAT3 inhibitor LLL3 was used to investigate the impact of STAT3 inhibition following OSM stimulation of OSA cells.</p> <p>Results</p> <p>Our data demonstrate that the OSM receptor (OSMR), but not IL-6 or its receptor, is expressed by all human and canine OSA cell lines and canine OSA tumor samples; additionally, OSM expression was noted in all tumor samples. Treatment of OSA cell lines with OSM induced phosphorylation of STAT3, Src, and JAK2. OSM stimulation also resulted in a dose dependent increase in MMP2 activity and VEGF expression that was markedly reduced following treatment with the small molecule STAT3 inhibitor LLL3. Lastly, OSM stimulation of OSA cell lines enhanced invasion through Matrigel, particularly in the presence of rhHGF. In contrast, both OSM and HGF stimulation of OSA cell lines did not alter their proliferative capacity.</p> <p>Conclusions</p> <p>These data indicate OSM stimulation of human and canine OSA cells induces STAT3 activation, thereby enhancing the expression/activation of MMP2 and VEGF, ultimately promoting invasive behavior and tumor angiogenesis. As such, OSM and its receptor may represent a novel target for therapeutic intervention in OSA.</p

Evaluation of Some Vulval Appendages in Nematode Taxonomy

A survey of the nature and phylogenetic distribution of nematode vulval appendages revealed 3 major classes based on composition, position, and orientation that included membranes, flaps, and epiptygmata. Minor classes included cuticular inflations, protruding vulvar appendages of extruded gonadal tissues, vulval ridges, and peri-vulval pits. Vulval membranes were found in Mermithida, Triplonchida, Chromadorida, Rhabditidae, Panagrolaimidae, Tylenchida, and Trichostrongylidae. Vulval flaps were found in Desmodoroidea, Mermithida, Oxyuroidea, Tylenchida, Rhabditida, and Trichostrongyloidea. Epiptygmata were present within Aphelenchida, Tylenchida, Rhabditida, including the diverged Steinernematidae, and Enoplida. Within the Rhabditida, vulval ridges occurred in Cervidellus, peri-vulval pits in Strongyloides, cuticular inflations in Trichostrongylidae, and vulval cuticular sacs in Myolaimus and Deleyia. Vulval membranes have been confused with persistent copulatory sacs deposited by males, and some putative appendages may be artifactual. Vulval appendages occurred almost exclusively in commensal or parasitic nematode taxa. Appendages were discussed based on their relative taxonomic reliability, ecological associations, and distribution in the context of recent 18S ribosomal DNA molecular phylogenetic trees for the nematodes. Characters were found to be distributed across subsets of terminal and phylogenetically distant taxa, demonstrating considerable homoplasy. Accurate definitions, terminology, and documentation of the taxonomic distribution of vulval appendages are important in evaluations of hypotheses for either parallelism and developmental constraint or convergence and adaptation