Reversible Disassembly of the Actin Cytoskeleton Improves the Survival Rate and Developmental Competence of Cryopreserved Mouse Oocytes

Effective cryopreservation of oocytes is critically needed in many areas of human reproductive medicine and basic science, such as stem cell research. Currently, oocyte cryopreservation has a low success rate. The goal of this study was to understand the mechanisms associated with oocyte cryopreservation through biophysical means using a mouse model. Specifically, we experimentally investigated the biomechanical properties of the ooplasm prior and after cryopreservation as well as the consequences of reversible dismantling of the F-actin network in mouse oocytes prior to freezing. The study was complemented with the evaluation of post-thaw developmental competence of oocytes after in vitro fertilization. Our results show that the freezing-thawing process markedly alters the physiological viscoelastic properties of the actin cytoskeleton. The reversible depolymerization of the F-actin network prior to freezing preserves normal ooplasm viscoelastic properties, results in high post-thaw survival and significantly improves developmental competence. These findings provide new information on the biophysical characteristics of mammalian oocytes, identify a pathophysiological mechanism underlying cryodamage and suggest a novel cryopreservation method

Haptoglobin concentration in peritoneal fluid of women with endometriosis

ABSTRACT: Objective: To determine the concentration of haptoglobin (Hp) in peritoneal fluid (PF) of women with and without endometriosis and its correlation with the phase of the menstrual cycle. The expression of Hp\u3b2e (determined by two-dimensional polyacrylamide gel electrophoresis) was correlated to total PF Hp concentration. Materials and methods: PF samples were obtained from 240 women with or without endometriosis. PF Hp concentration was measured by a sandwich enzyme-linked immunosorbent assay. Hp\u3b2e expression in PF was estimated by two-dimensional polyacrylamide gel electrophoresis. Results: PF Hp concentration was similar in women with and without endometriosis; it was significantly higher during the luteal phase of the menstrual cycle than in the follicular phase in control subjects, but not in women with endometriosis. There was a positive correlation between serum progesterone level and PF Hp concentration in control subjects during the luteal phase of the menstrual cycle. There was a mild positive correlation between PF Hp concentration and Hp\u3b2e expression in controls but not in wom- en with endometriosis. PF Hp concentration was not correlated with the ASRM stage of endometriosis and the type of endometriotic lesions. Conclusions: Women with and without endometriosis have similar PF Hp concentration. There is a mild positive correlation between PF Hp concentration and Hp\u3b2e expression in control subjects but not in women with endometriosis. Future investigations should aim to produce antibodies against specific epitopes of Hp\u3b2e that might allow its measurement by ELISA

First trimester maternal serum concentrations of fetal antigen 2 in normal pregnancies and those affected by trisomy 21

Serum concentrations of fetal antigen 2 (FA-2), the amino-propeptide of the al chain of collagen type I, were measured in peripheral blood from women with normal (n = 234) and trisomy 21 affected (n = 14) pregnancies between 9 and 11 weeks gestation. Serum FA-2 concentrations were seen to be stable throughout this period, and though raised FA-2 concentrations were seen at the 10th week of gestation, a statistically significant difference between normal and trisomy 21 affected pregnancies was not found overall. Therefore it seems unlikely that FA-2 has a role in first trimester screening for trisomy 21, despite the fact that significantly higher FA-2 concentrations in trisomy 21 and significantly lower concentrations in trisomy 18 had been previously demonstrated in amniotic fluid in the second trimester

GnRH analogue remarkably down-regulates inflammatory proteins in peritoneal fluid proteome of women with endometriosis.

OBJECTIVE: To determine the changes in the peritoneal fluid proteome of women with endometriosis determined by the administration of gonadotropin-releasing hormone analogues (GnRH-a). STUDY DESIGN: Peritoneal fluid samples were collected during laparoscopy from patients under GnRH-a and from women who did not receive any type of hormonal treatment in the 6 months before surgery. Samples were subjected to 2-D gel electrophoresis and compared by computerized analysis. Protein spots differentially expressed between the study groups were identified by liquid chromatography tandem mass spectrometry. RESULTS: More than 470 protein spots were analyzed. Several proteins with significant alterations were found. The down-regulated molecules were isoforms of alpha 2-HS glycoprotein, alpha 1-antitrypsin, S100-A8, haptoglobin alpha chain and vitamin D-binding protein. No protein spot had significantly higher expression in peritoneal fluid of women under GnRH-a than in untreated patients. CONCLUSION: Several inflammatory molecules present in peritoneal fluid are down-regulated during treatment with GnRH-a; administration of this drug reduces the inflammation in the peritoneal cavity