Expression of Cu/Zn and Mn superoxide dismutases during bovine embryo development: influence of in vitro culture.

peer reviewedTemporal pattern of expression of Cu/Zn and Mn superoxide dismutases (SODs) was investigated in bovine oocytes and embryos produced in vitro in two different culture conditions and in vivo after superovulation. SODs were examined at a transcriptional level in single oocytes and embryos by reverse transcriptase-polymerase chain reaction (RT-PCR) and, at a protein level, by Western blotting on pools of embryos. mRNA encoding Cu/Zn SOD were detected in in vitro bovine embryos throughout preattachment development as well as in in vivo derived morulae and blastocysts. Transcripts for Mn SOD gene were detected in most immature and in vitro matured oocytes as well as in some zygotes and 5- to 8-cell embryos while no transcript was found at the 9-to 16-cell stage in both culture conditions. In vitro embryonic expression of Mn SOD was detected earlier in the presence of serum. Half of the morulae showed the transcript if cultured with 5% serum while none without serum. At the blastocyst stage Mn SOD could be detected independently of culture conditions. For in vivo-derived embryos Mn SOD transcripts were detected both in morulae and blastocysts. Immunoblotting analyses revealed that Cu/Zn SOD and Mn SOD were also present at a protein level in in vitro-derived zygotes and blastocysts. Together these data demonstrate, for the first time, that Mn SOD is transcribed and that Cu/Zn and Mn SOD proteins are expressed in preimplantation bovine embryos. Finally, they suggest that Mn SOD transcription is altered by in vitro culture conditions

Caractérisation d'un système NADPH/thioredoxine non chloroplastique et de thioredoxines chloroplastiques de type f et m dans l'Acetabularia mediterranea: Epreuve du rôle possible des thiols dans la morphogenèse de l'algue

Doctorat en Sciencesinfo:eu-repo/semantics/nonPublishe

Typical and subtle atypical presentations of endometriosis.

PURPOSE OF REVIEW: To review the concept of typical, subtle and invisible endometriosis and analyze the evolution and progression of the disease, as well as some factors possibly involved in the pathogenesis. RECENT FINDINGS: Although rejected by Redwine, the concept of non-visible endometriosis has been proven to exist. Some new ideas on the implication of reactive oxygen species in the development of endometriosis and its early stages are described here. SUMMARY: The increased diagnosis of endometriosis can be explained not only by the increased experience and ability of the surgeon to detect typical and non-typical endometriotic lesions, but also by a better understanding of the pathogenesis

Oxidative stress and peritoneal endometriosis.

OBJECTIVE: To review the literature associating pelvic endometriosis with oxidative stress and to discuss the potential causes and consequences of a pro-oxidant environment in the peritoneal cavity. DESIGN: Literature survey. RESULT(S): Several studies suggest that oxidative stress is a component of the inflammatory reaction associated with endometriosis. Evidence includes the prevention of endometriosis induction in rabbits by the addition of antioxidants, an increase in reactive oxygen species release by macrophages, increased peritoneal levels of oxidized low-density lipoproteins and their by-products, altered expression of endometrial pro-oxidant and antioxidant enzymes, and consumption of peritoneal fluid vitamin E. Retrograde menstruation is likely to carry highly pro-oxidant factors, such as heme and iron, into the peritoneal cavity, as well as apoptotic endometrial cells, which are well-known inducers of oxidative stress. Reactive oxygen species may be involved in endometriosis-associated infertility and may play a role in the regulation of the expression of genes encoding immunoregulators, cytokines and cell adhesion molecules implicated in the pathogenesis of endometriosis. CONCLUSION(S): Better understanding of the mechanisms of reactive oxygen species production and detoxification and further investigation of their effect on the peritoneal environment are essential to obtain new insight into this disease and eventually develop new diagnostic and therapeutic strategies

[Calves Born After Transfer of Unfrozen and Frozen-thawed In-vitro Produced Embryos]

A total of 139 embryos produced in vitro either in coculture with oviduct cells or in medium conditioned by these cells in the presence or absence of serum were transferred either after culture (63) or after culture and freezing (76) resulting in 29 calves born (18 males, 11 females) weighting between 40 and 60 kg, after a mean gestation length of 290 days. The bull used for fertilization was of the Belgian Blue breed and his gestation length recorded in vivo is longer than the mean. The final calving yield was 21 %. The best calving rates were obtained from embryos cultured in 'serum-free' oviduct-conditioned medium 5/16 (31 %) for embryos transferred individually after culture and 4/17 (24 %) for embryos transferred after culture and freezing. Too more pregnancies are confirmed in the last group after transfer of 2 day 6 blastocysts stored for 1 year in liquid nitrogen. Serum PAG (Pregnancy Associated Glycoprotein) level was lower in the recipients receiving frozen embryos than in those receiving non frozen embryos. A field trial consisting in the transfer of 105 frozen-thawed embryos by Al veterinarians resulted in 12 pregnancies (11 %)

Iron overload in the peritoneal cavity of women with pelvic endometriosis.

OBJECTIVE: To examine the possible involvement of iron in the physiopathology of endometriosis. DESIGN: Prospective study. SETTING: Department of gynecology in a university hospital. PATIENT(S): Seventy patients undergoing laparoscopy. INTERVENTION(S): Collection of peritoneal fluid (n = 57), blood samples, and biopsy samples from endometrium (n = 62) and from endometriotic (n = 33) and normal-appearing peritoneum (n = 53). MAIN OUTCOME MEASURE(S): Measurement of iron and ferritin in serum and peritoneal fluid and staining of iron deposits with Prussian blue in tissues. RESULT(S): Iron and ferritin concentrations were significantly higher in the peritoneal fluid of patients with endometriosis compared with controls during the secretory phase. Higher rates of ferritin and hemosiderin deposits were observed in the peritoneum adjacent to red (100%), black (57%), and white (62%) lesions compared with normal-appearing peritoneum (25%). Deposits were more frequent during the secretory phase than the proliferative phase in healthy peritoneum from controls, whereas they were found throughout the cycle in the vicinity of lesions in patients with endometriosis. Similar rates of iron deposition were observed in the stroma of black and white lesions and in eutopic endometrium from patients with endometriosis. CONCLUSION(S): Iron overload was observed in the cellular and peritoneal fluid compartments of the peritoneal cavity of women with endometriosis. Iron deposits seem to be related to the presence of lesions, suggesting that iron may be involved in the pathogenesis of endometriosis

Factors Affecting Bovine Embryo Development in Synthetic Oviduct Fluid Following Oocyte Maturation and Fertilization In-vitro

Employing a total of 3465 bovine oocytes this study was aimed at improving the efficiency of bovine embryo production under defined and undefined conditions. Following in vitro maturation (IVM) and in vitro fertilization (IVF), oocytes were allocated to various culture treatments using synthetic oviduct fluid (SOF). In our 3 experiments we showed that : 1) the addition of fetal calf serum (FCS 10% v/v) to SOF droplets after 20 to 24 h significantly improved blastocyst yields on Day 6 (21 vs 12%; P<0.01), but not at later stages, and resulted in significantly higher Day-8 blastocyst cell numbers (148+/-61 vs 92+/-35 P<0.05); 2) the removal of bovine serum albumin (BSA) from the standard SOF medium resulted in significantly reduced blastocyst yields on Days 6, 7 and 8, respectively (17 vs 8%; 28 vs 18%; 31 vs 21%; P<0.05); 3) the presence or absence of cumulus cells surrounding the presumptive zygote in culture in SOF had no effect on cleavage rate, percentage of 5-8 cell embryos or blastocyst yields (Day 6, 7 or 8); 4) the culture of presumptive zygotes in SOF in an atmosphere of 5% CO2 in air (20% O-2) resulted in significantly reduced development compared with culture in 5% CO2, 5%O-2, 90% N-2 in terms of blastocyst yield on Days 6, 7 and 8 and on Day 8 hatching rate, respectively (5 vs 22%; 9 vs 33%; 13 vs 48%; 50 vs 8%; P<0.001) and 5) embryo density (1 embryo per 1 or 3 mu l SOF) or replacing the culture medium every 48 h had no effect when SOF was supplemented with serum; however, under serum-free conditions, changing of the media resulted in a slightly improved Day-6 blastocyst yield such that renewal of serum-free medium mimicked the effect of serum addition

Effect of estrus-associated glycoprotein and tissue inhibitor of metalloproteinase-1 secreted by oviduct cells on in vitro bovine embryo development.

The effect of two glycoproteins (estrus-associated glycoprotein [EGP] and tissue inhibitor of metalloproteinase [TIMP-1]) secreted by bovine oviduct cells on in vitro bovine embryo development was assessed. A first set of experiments was conducted to determine whether the embryotrophic activity of the bovine oviduct-conditioned medium (BOCM) was correlated with the presence of EGP or TIMP-1. EGP and TIMP-1 were detected in BOCM, supporting the development of 22% zygotes to the blastocyst stage, as well as in BOCM yielding a low blastocyst rate (3-4% blastocysts). These glycoproteins do not seem to be necessary for bovine embryo development up to the blastocyst stage in our BOCM. In a second experiment, zygotes were cultured in modified synthetic oviduct fluid (mSOF) supplemented with different concentrations (0.5, 5, 50, and 500 micrograms/ml) of purified bovine EGP. In the third experiment, since purified bovine TIMP-1 was not available, zygotes were cultured in BOCM depleted of TIMP-1 by immunoprecipitation treatment. Adding EGP to mSOF, or removing TIMP-1 from BOCM, did not affect bovine embryo development up to the blastocyst stage, or mean number of cells per blastocyst after 8 days of culture. The results indicate that, under our culture conditions, EGP and TIMP-1 do not play an important role in sustaining bovine embryo development, and do not influence blastocyst quality, assessed in terms of total number of cells per embryo