No abstract

Sperm DNA fragmentation may play a role in unexplained RPL despite normal semen analysis parameters.

Objective-We previously demonstrated that adherence of endometrial epithelial (EECs) and stromal cells (ESCs) to peritoneal mesothelial cells (PMCs) is partly regulated by ESC/EEC CD44 interactions with PMC associated hyaluronan. CD44, a transmembrane glycoprotein and major ligand for hyaluronan, has numerous splice variants which may impact hyaluronan binding. Here, we assessed whether ESCs and EECs from women with endometriosis demonstrate increased adherence to PMCs and examined CD44 splice variants' potential role in this process. Design-In vitro study. Setting-Academic medical Center Patient(s)-Fertility patients with and without endometriosisIntervention(s)-Menstrual endometrium was collected from women with and without endometriosis confirmed surgically. The adherence of ESC/EECs to PMCs was measured. ESC/EEC CD44 splice variants were assessed using dot blot analysis. Results-ESCs and EECs from women with endometriosis demonstrated increased adherence toPMCs. The predominant CD44 splice variants expressed by ESCs and EECs from women with and without endometriosis were v3, v6, v7, v8, v9, and v10. ESCs and EECs from women with endometriosis were more likely to express v6, v7, v8 or v9.Conclusions-Increased eutopic endometrial-PMC adherence and CD44 splice variant expression may contribute to the histogenesis of endometriotic lesions. Elucidation of factors controlling this expression may lead to novel endometriosis therapies.

Beta 1 and beta 2 are the designations given to two forms of beta-tubulin that have different electrophoretic mobilities on discontinuous polyacrylamide gels in the presence of sodium dodecyl sulfate [Little, M. (1979) FEBS Lett. 108, 283-286]. Beta 1 and beta 2 constitute respectively 75% and 25% of the total beta-tubulin in bovine brain. Although beta 1 appears to be ubiquitous in animals, beta 2 has so far only been found in the brains of cows, pigs, deer, rats, chicks, and dogfish but not in squid brain. Beta 2 is not found in bovine kidneys, in porcine lungs, or in any nonchordate tubulin that has been examined. When tubulin is reacted with the sulfhydryl-directed reagent N,-N'-ethylenebis(iodoacetamide) (EBI), beta 1, but not beta 2, is converted to a faster moving form, beta. The yield of beta 2 in this reaction is not altered by the presence of drugs. When [14C]EBI is used as a probe, most of the label is incorporated into beta 1 rather than beta 2. Tubulin molecules that have reacted with EBI to form beta are much less likely to polymerize into microtubules than are molecules that have not formed beta. In view of the observation that only beta 1, and not beta 2, can form beta, it is possible that beta 1 represents a form of tubulin whose assembly may be regulated by a mechanism involving sulfhydryls. In contrast, beta 2 may represent a form of tubulin whose assembly is regulated by some other mechanism.

Transforming growth factor beta 1 (TGF-beta1) levels are increased in the peritoneal fluid of endometriosis patients, and endometrial cells express TGF-beta signaling components; however, little is known regarding the role of TGF-beta in endometriosis. Our objective was to examine the effects of TGF-beta1 on (i) the expression of macrophage colony-stimulating factor receptor encoded by the c-fms gene, (ii) transmesothelial invasiveness of endometrial cells, (iii) cellular proliferation and (iv) attachment to peritoneal mesothelial cells (PMCs). Effects of TGF-beta1 on c-fms mRNA expression were determined by real-time RT-PCR and c-fms cell-surface expression by flow cytometry. Effects of TGF-beta1 on the invasiveness of the immortalized endometrial epithelial cell (EEC) line EM42 and primary EECs were examined using a three-dimensional in vitro system modeling the peritoneum. Cellular proliferation and attachment to PMCs were also examined using established techniques. TGF-beta1 had little or no effect on cellular proliferation and endometrial cell attachment to PMCs. TGF-beta1 significantly induced the expression of c-fms mRNA and c-fms cell-surface expression. TGF-beta1 enhanced transmesothelial invasion by EM42 cells and EECs. Antagonists of TGF-beta1 signaling significantly inhibited both the induction of c-fms expression and cellular invasiveness, suggesting that additional studies are warranted to assess the therapeutic potential of TGF-beta antagonists in endometriosis.