Role of EGF on in situ culture of equine preantral follicles and metabolomics profile.

The effects of epidermal growth factor (EGF) concentrations (0, 10, 50, and 100ng/ml) on in vitro culture (IVC) of equine preantral follicles were evaluated using histology, estradiol and reactive oxygen species (ROS) production and metabolomics. After IVC, the percentage of normal follicles was lower (P<0.05) for all treatments when compared to non-cultured control. EGF 50ng/ml treatment had more (P<0.05) normal follicles at Day 7 of culture when compared with EGF 0 and 100ng/ml. EGF 50ng/ml had more (P<0.05) developing follicles than the 0ng/ml and 10ng/ml EGF treatments. Follicular and oocyte diameters were greater (P<0.05) with EGF 50ng/ml than the other cultured treatments, but similar (P>0.05) to the non-cultured control. From Day 1 to Day 7 estradiol production increased (P<0.05) in all EGF treatments. EGF 50ng/ml was the only treatment that maintained ROS production through IVC. Metabolomics profiles of the spent media indicated that eleven ions from variable influence in the projection (VIP) scores were higher represented in the EGF 50ng/ml treatment. In conclusion, EGF 50ng/ml treatment maintained follicle survival and ROS production, and promoted activation of cultured equine preantral follicles enclosed in ovarian tissue.

In vitro growth and maturation of isolated caprine preantral follicles: Influence of insulin and FSH concentration, culture dish, coculture, and oocyte size on meiotic resumption.

The aims of this study were: (1) to evaluate the effect of different insulin concentrations, alone or in combination with either a fixed FSH concentration or increasing FSH concentrations on the in vitro culture of isolated caprine preantral follicles and (2) to analyze the efficiency of two IVM media and maturation culture systems (with or without coculture with in vivo grown oocytes) on the meiosis resumption. Secondary follicles were cultured for 18 days in a basic medium supplemented with low- or high-insulin concentration alone or with a fixed FSH concentration or with increasing FSH concentrations. Oocytes grown in vivo or in vitro were matured alone or cocultured. The high-insulin concentration associated with fixed FSH treatment had higher meiotic resumption rate (P < 0.05) and was the only treatment capable of producing oocytes in metaphase II. The rates of germinal vesicle, germinal vesicle breakdown, metaphase I, metaphase II (MII), meiotic resumption, and oocyte diameter were similar between the maturation media. In conclusion, a basic medium supplemented with 10-µg/mL insulin and 100-µg/mL FSH throughout the culture period improved meiotic resumption rate and produced MII oocytes from caprine preantral follicles cultured in vitro. The MII rate was similar between in vivo and in vitro grown oocytes ≥110 µm.

Chemical composition of lipids present in cat and dog oocyte by matrix-assisted desorption ionization mass spectrometry (MALDI- MS).

The aim of the present study was to investigate the level of information on the chemical structures and relative abundances of lipids present in cat and dog oocytes by matrix-assisted laser desorption mass spectrometry (MALDI-MS). The MALDI-MS approach requires a simple analysis workflow (no lipid extraction) and few samples (two or three oocytes per analysis in this work) providing concomitant profiles of both intact phospholipids such as sphingomyelins (SM) and phosphatidylcholines (PC) as well as triacylglycerols (TAG). The lipids were detected in oocytes by MALDI using dihydroxybenzoic acid (DHB) as the matrix. The most abundant lipid present in the MS profiles of bitch and queen oocytes was a PC containing 34 carbons and one unsaturation [PC (34:1)]. Oocytes of these two species are characterized by differences in PC and TAG profiles detected qualitatively as well as by means of principal component analysis (PCA). Cat oocytes were mainly discriminated by more intense C52 and C54 TAG species and a higher number of unsaturations, indicating predominantly linoleic and oleic fatty acyl residues. Comparison of the lipid profile of bitch and queen oocytes with that of bovine oocytes revealed some similarities and also some species specificity: TAG species present in bovine oocytes were also present in bitches and queens; however, a more pronounced contribution of palmitic, stearic and oleic fatty acid residues was noticed in the lipid profile of bovine oocytes. MALDI-MS provides novel information on chemical lipid composition in canine and feline oocytes, offering a suitable tool to concomitantly monitor, in a nearly direct and simple fashion the composition of phospholipids and TAG. This detailed information is highly needed to the development of improved protocols for in vitro culture and cryopreservation of cat and dog oocytes.

PP098. Lipidic fingerprinting in women with early-onset preeclampsia: A first look.

INTRODUCTION: preeclampsia is characterized by intense inflammatory response and an anti-angiogenic state. Maternal obesity has been considered to have important impact on the genesis of preeclampsia as lipotoxicity leads to maternal endothelial dysfunction and chronic inflammation. Here we investigate the plasma lipid profile of preeclamptic women. OBJECTIVES: identify possible lipid biomarkers for preeclampsia. METHODS: this study included 8 pregnant women with early-onset preeclampsia (before 34weeks gestation) and 8 normal pregnant women. Each patient in the preeclampsia group was matched to a patient in the control group according to gestational age at the time of sample collection. All patients in the control group were followed until term and had normal outcomes. To investigate the lipid profile, lipids were extracted from plasma samples using the Bligh-Dyer protocol and the extracts were subjected to MALDI-TOF Mass Spectrometry. Data matrix was exported for partial least squares discriminant analysis. All the variables analysed were sorted by a score number named Variable Importance in the Projection. The major discriminant variables were selected and underwent to Mann-Whitney U test. RESULTS: a total of 1290 ions were initially identified during lipidomic assessment. Twelve m/z signals were highlighted as the most important lipids for the discrimination of patients with preeclampsia. The identification of these differential lipids was carried out through Lipid Database Search. The main classes identified were Glycerophosphocholines [GP01], Glycerophosphoserines [GP03], Glycerophosphoglycerols [GP04], Glycosyldiradylglycerols [GL05] and Glycerophosphates [GP10]. CONCLUSION: Our results suggest that some lipid species may be potential biomarkers for early-onset preeclampsia.

Interrupcion de la vena cava inferior en la prevencion del embolismo pulmonar. / The inferior vena cava interruption in the prevention of pulmonary embolism. Comments on 59 cases

Se presenta una experiencia de 8 anos en la interrupcion de la vena cava inferior para la prevencion del embolismo pulmonar.Se cotejan los resultados de 24 procedimientos quirurgicos y 38 protesis de Mobin Uddin. La mortalidad operatoria fue superior en el procedimiento quirurgico, 25%, contra el filtro. El edema residual y el embolismo recurrente, no presentaron diferencias significativas en los dos metodos.Se hacen consideraciones sobre los procedimientos de obliteracion total y parcial lo mismo que con los procedimientos quirurgicos y por cateterismo

Interrupcion de la vena cava inferior en la prevencion del embolismo pulmonar. / The inferior vena cava interruption in the prevention of pulmonary embolism. Comments on 59 cases

Se presenta una experiencia de 8 anos en la interrupcion de la vena cava inferior para la prevencion del embolismo pulmonar.Se cotejan los resultados de 24 procedimientos quirurgicos y 38 protesis de Mobin Uddin. La mortalidad operatoria fue superior en el procedimiento quirurgico, 25%, contra el filtro. El edema residual y el embolismo recurrente, no presentaron diferencias significativas en los dos metodos.Se hacen consideraciones sobre los procedimientos de obliteracion total y parcial lo mismo que con los procedimientos quirurgicos y por cateterismo