Oxidative dtress status and placental implications in diabetic rats undergoing swimming exercise after embryonic implantation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of FGF10 on bovine oocyte meiosis progression, apoptosis, embryo development and relative abundance of developmentally important genes in vitro

Contents Fibroblast growth factor (FGF10) acts at the cumulus oocyte complex, increasing the expression of cumulus cell expansion-related genes and oocyte competency genes. We tested the hypothesis that addition of FGF10 to the maturation medium improves oocyte maturation, decreases the percentage of apoptotic oocytes and increases development to the blastocyst stage while increasing the relative abundance of developmentally important genes (COX2, CDX2 and PLAC8). In all experiments, oocytes were matured for 22h in TCM-199 supplemented with 0, 2.5, 10 or 50ng/ml FGF10. In Experiment 1, after maturation, oocytes were stained with Hoechst to evaluate meiosis progression (metaphase I, intermediary phases and extrusion of the first polar body) and submitted to the TUNEL assay to evaluate apoptosis. In Experiment 2, oocytes were fertilized and cultured to the blastocyst stage. Blastocysts were frozen for analysis of COX2, CDX2 and PLAC8 relative abundance. In Experiment 1, 2.5ng/ml FGF10 increased (p<0.05) the percentage of oocytes with extrusion of the first polar body (35%) compared to 0, 10 and 50ng/ml FGF10 (21, 14 and 12%, respectively) and FGF10 decreased the percentage of oocytes that were TUNEL positive in all doses studied. In Experiment 2, there was no difference in the percentage of oocytes becoming blastocysts between treatments and control. Real-time RT-PCR showed a tendency of 50ng/ml FGF10 to increase the relative abundance of COX2 and PLAC8 and of 10ng/ml FGF10 to increase CDX2. In conclusion, the addition of FGF10 to the oocyte maturation medium improves oocyte maturation in vitro, decreases the percentage of apoptotic oocytes and tends to increase the relative abundance of developmentally important genes.

The interval between the emergence of pharmacologically synchronized ovarian follicular waves and ovum pickup does not significantly affect in vitro embryo production in Bos indicus, Bos taurus, and Bubalus bubalis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Paradoxical effects of bovine somatotropin treatment on the ovarian follicular population and in vitro embryo production of lactating buffalo donors submitted to ovum pick-up

The aim of the present study was to evaluate the effect of bovine somatotropin (bST; 500 mg) administration on lactating buffalo donors submitted to two different ovum pick-up (OPU) and in vitro embryo production schemes with a 7 or 14 d intersession OPU interval. A total of 16 lactating buffalo cows were randomly assigned into one of four experimental groups according to the bST treatment (bST or No-bST) and the OPU intersession interval (7 or 14 d) in a 2 x 2 factorial design (16 weeks of OPU sessions). The females submitted to OPU every 14d had a larger (P < 0.001) number of ovarian follicles suitable for puncture (15.6 +/- 0.7 vs. 12.8 +/- 0.4) and an increased (P = 0.004) number of cumulus-oocyte complexes (COCs) recovered (10.0 +/- 0.5 vs. 8.5 +/- 0.3) compared to the 7 d interval group. However, a 7 or 14 d interval between OPU sessions had no effect (P = 0.34) on the number of blastocysts produced per OPU (1.0 +/- 0.1 vs. 13 +/- 0.2, respectively). In addition, bST treatment increased (P < 0.001) the number of ovarian follicles suitable for puncture (15.3 +/- 0.5 vs. 12.1 +/- 0.4) but reduced the percentage (18.9% vs. 10.9%; P = 0.009) and the number (1.4 +/- 0.2 vs. 0.8 +/- 0.1; P = 0.003) of blastocysts produced per OPU session compared with the non-bST-treated buffaloes. In conclusion, the 14d interval between OPU sessions and bST treatment efficiently increased the number of ovarian follicles suitable for puncture. However, the OPU session interval had no effect on embryo production, and bST treatment reduced the in vitro blastocyst outcomes in lactating buffalo donors.

Expression studies in the embryo and in the micropylar endosperm of germinating coffee (Coffea arabica cv. Rubi) seeds

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Manifestation of estrous behavior and subsequent progesterone concentration at timed-embryo transfer in cattle are positively associated with pregnancy success of recipients

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effect of cell cycle synchronization on the accuracy of murine and bovine embryo sex determination

Different cell cycle synchronization methods were used to increase the mitotic index and accuracy of sex determination in murine and bovine embryos. For sexing purposes, colchicine treatment for 2, 4, 6 and 8 h and the FdU-thymidine-colchicine combination were tested in murine embryos. The best results were obtained with colchicine treatment for 8 h (96.88% accuracy) and with FdU-thymidine-colchicine (97.22% accuracy). Mitotic indexes differed significantly between the 2 treatments (21.71% for colchicine and 32.95% for FdU-thymidine-colchicine). For sex identification of murine and bovine demi-embryos, both treatments were demonstrated to be equally effective (nearly 90%). The mitotic index for the FdU-treated murine demi-embryos (19.04%) was higher than the one obtained for the 8-h colchicine treatment (15.62%).

Predicting embryo presence and viability

Pregnancy establishment, followed by birth of live offspring, is essential to all mammals. The biological processes leading up to pregnancy establishment, maintenance, and birth are complex and dependent on the coordinated timing of a series of events at the molecular, cellular, and physiological level. The ability to ovulate a competent oocyte, which is capable of undergoing fertilization, is only the initial step in achieving a successful pregnancy. Once fertilization has occurred and early embryonic development is initiated, early pregnancy detection is critical to provide proper prenatal care (humans) or appropriate management (domestic livestock). However, the simple presence of an embryo, early in gestation, does not guarantee the birth of a live offspring. Pregnancy loss (embryonic mortality, spontaneous abortions, etc.) has been well documented in all mammals, especially in humans and domestic livestock species, and is a major cause of reproductive loss. It has been estimated that only about 25-30 % of all fertilized oocytes in humans result in birth of a live offspring; however, identifying the embryos that will not survive to parturition has not been an easy task. Therefore, investigators have focused the identification of products in maternal circulation that permit the detection of an embryo and assessment of its well-being. This review will focus on the advances in predicting embryonic presence and viability, in vivo.

Single-cell expression profiling reveals a dynamic state of cardiac precursor cells in the early mouse embryo

In the early vertebrate embryo, cardiac progenitor/precursor cells (CPs) give rise to cardiac structures. Better understanding their biological character is critical to understand the heart development and to apply CPs for the clinical arena. However, our knowledge remains incomplete. With the use of single-cell expression profiling, we have now revealed rapid and dynamic changes in gene expression profiles of the embryonic CPs during the early phase after their segregation from the cardiac mesoderm. Progressively, the nascent mesodermal gene Mesp1 terminated, and Nkx2-5+/Tbx5+ population rapidly replaced the Tbx5low+ population as the expression of the cardiac genes Tbx5 and Nkx2-5 increased. At the Early Headfold stage, Tbx5-expressing CPs gradually showed a unique molecular signature with signs of cardiomyocyte differentiation. Lineage-tracing revealed a developmentally distinct characteristic of this population. They underwent progressive differentiation only towards the cardiomyocyte lineage corresponding to the first heart field rather than being maintained as a progenitor pool. More importantly, Tbx5 likely plays an important role in a transcriptional network to regulate the distinct character of the FHF via a positive feedback loop to activate the robust expression of Tbx5 in CPs. These data expands our knowledge on the behavior of CPs during the early phase of cardiac development, subsequently providing a platform for further study.

Lipid structural information from a single equine embryo by MALDI-MS

O objetivo deste trabalho foi relatar o potencial da técnica de MALDI-MS para caracterizar espécies de lipídios presentes em um único embrião equino e estudar algumas estruturas lipídicas detectadas por dissociação induzida por colisão (CID). No espectro de modo íon positivo, pudemos observar espécies, principalmente, protonadas e sodiadas de esfingomielinas (SM), fosfatidileolinas (PC) e triacilgliceróis (TAG). No modo negativo, observamos fosfatidiletanolaminas (PE) e fosfatidilinositos (PI). Espectros de íons de lípidos com maior intensidade foram utilizados para demonstrar o potencial da informação estrutural por MALDI-MS/MS. O espectro no modo positivo de m/z (massa sobre carga) 760,6 (atribuída como PC34:1) apresentou características de fragmentos PC de m/z 184,1 (denominada cabeça polar de colina), além de perda neutral (NL) de m/z 183 (fosforilcolina). Para o íon de m/z 766,6 (atribuída como PE38:5), observou-se a NL de 140, característica do PE. Para o íon de m/z 808,7 (38,5 atribuído como PC), além do fragmento m/z 184,1 na NL de 183, foi possível observar a perda de trimetilamina (íon de m/z 749,6) e o ciclofosfano (íon de m/z 147,0). Finalmente, para o modo de íon negativo, foram isolados e fragmentados o íon de m/z 863,6 que foi atribuído como PI36:1, devido à presença de m/z 153 (fosfato de glicerol – H2O-H ), 223 (inositol fosfo - 2H2O-H) , 241 (fosfoinositol – H2O-H), 281 (ácido oleico) e 581,3 (lisofosfoinositol – H2O+H). Concluímos que a MALDI - MS permite a detecção de uma ampla gama de espécies de PC, SM, PE, PI e TAG lipídicas, bem como a caracterização rápida e confiante de estruturas lipídicas a partir de um único embrião equino.

Effect of antioxidants during bovine in vitro fertilization procedures on spermatozoa and embryo development

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Visual outcomes and pacient satisfaction follwing implantation of a supplementary multifocal IOL in patients undergoing cataract surgery

Purpose: To assess visual outcomes and patient satisfaction following implantation of the Sulcoflex® multifocal intraocular lens (IOL; Rayner Intraocular Lenses Ltd., Hove, UK) in a procedure combining capsular bag lens implantation with sulcus placement of the Sulcoflex® IOL. Setting: Instituto de Oftalmologia de Assis, Assis, SP, Brazil. Methods: Cataract patients > 45 years, with hyperopia ≥ 1.50 D and potential acuity measurement ≥ 20/30 undergoing Sulcoflex® multifocal IOL implantation were included. Monocular and binocular uncorrected near and distance visual acuity (VA) were evaluated at five days, one month, and three months postoperatively. Contrast sensitivity and refraction were measured in a subset of patients three months postoperatively. Patient satisfaction was assessed one month postoperative. Results: This non-consecutive case series comprised 25 eyes of 13 patients. Eleven eyes (52%) had pre-existing retinal pathologies. Monocular distance VA improved significantly at all follow-up visits. At final follow-up, 88% of eyes had monocular uncorrected distance VA (UDVA) of at least 20/25 and 24% had monocular UDVA of 20/20. All eyes had binocular UDVA of at least 20/25, and 58% had binocular UDVA of 20/20. Monocular uncorrected near vision (UNVA) was J1 in 68% of eyes and all patients had binocular UNVA of J1. Of all eyes studied, 92% and 58% achieved a spherical equivalent within 1 D and −0.5 D, respectively. The majority of patients reported satisfaction with visual outcomes. Complications included a postoperative intraocular pressure spike in four eyes. Conclusion: The Sulcoflex® multifocal IOL improves near and distance VA in cataract patients with retinal abnormalities and good VA potential.

Implantation of Subcutaneous Radio Transmitters in the Harbor Seal (Phoca vitulina)

Radio telemetry has become a standard tool for studying the behavior, physiology, life history traits, and population dynamics of marine mammals. Radio transmitters typically are attached to the hind flippers of pinnipeds or glued to the fur using marine epoxy or other cyanocrylare adhesives (Fedak et al. 1983, Bengtson 1993, Jeffries et al. 1993). Longterm data acquisition is difficult, however, because radio-flipper transmitters commonly tear from the webbing of the flipper and instruments that are glued to the fur are shed during the seasonal molt.

Modulation transfer function and optical quality after bilateral implantation of a+3.00 D versus a+4.00 D multifocal intraocular lens

PURPOSE: To determine whether the improvement in intermediate vision after bilateral implantation of an aspheric multifocal intraocular lens (IOL) with a +3.00 diopter (D) addition (add) occurs at the expense of optical quality compared with the previous model with a +4.00 D add. SETTING: Department of Ophthalmology, University of Sao Paulo, Sao Paulo, Brazil. DESIGN: Prospective randomized double-masked comparative clinical trial. METHODS: One year after bilateral implantation of Acrysof Restor SN6AD1 +3.00 D IOLs or Acrysof Restor SN6AD3 +4.00 D IOLs, optical quality was evaluated by analyzing the in vivo modulation transfer function (MTF) and point-spread function (expressed as Strehl ratio). The Strehl ratio and MTF curve with a 4.0 pupil and a 6.0 mm pupil were measured by dynamic retinoscopy aberrometry. The uncorrected and corrected distance visual acuities at 4 m, uncorrected and distance-corrected near visual acuities at 40 cm, and uncorrected and distance-corrected intermediate visual acuities at 50 cm, 60 cm, and 70 cm were measured. RESULTS: Both IOL groups comprised 40 eyes of 20 patients. One year postoperatively, there were no statistically significant between-group differences in the MTF or Strehl ratio with either pupil size. There were no statistically significant between-group differences in distance or near visual acuity. Intermediate visual acuity was significantly better in the +3.00 D IOL group. CONCLUSION: Results indicate that the improvement in intermediate vision in eyes with the aspheric multifocal +3.00 D add IOL occurred without decreasing optical quality over that with the previous version IOL with a +4.00 D add.

Transcatheter aortic valve implantation with a self-expanding nitinol bioprosthesis: Prediction of the Need for Permanent Pacemaker Using Simple Baseline and Procedural Characteristics

Objective: To ascertain incidence and predictors of new permanent pacemaker (PPM) following transcatheter aortic valve implantation (TAVI) with the self-expanding aortic bioprosthesis. Background: TAVI with the Medtronic Corevalve (MCV) Revalving System (Medtronic, Minneapolis, MN) has been associated with important post-procedural conduction abnormalities and frequent need for PPM. Methods: Overall, 73 consecutive patients with severe symptomatic AS underwent TAVI with the MCV at two institutions; 10 patients with previous pacemaker and 3 patients with previous aortic valve replacement were excluded for this analysis. Clinical, echocardiographic, and procedural data were collected prospectively in a dedicated database. A standard 12-lead ECG was recorded in all patients at baseline, after the procedure and predischarge. Decision to implant PPM was taken according to current guidelines. Logistic multivariable modeling was applied to identify independent predictors of PPM at discharge. Results: Patients exhibited high-risk features as evidenced by advanced age (mean = 82.1 +/- 6.2 years) and high surgical scores (logistic EuroSCORE 23.0 +/- 12.8%, STS score 9.4 +/- 6.9%). The incidence of new PPM was 28.3%. Interventricular septum thickness and logistic Euroscore were the baseline independent predictors of PPM. When procedural variables were included, the independent predictors of PPM were interventricular septum thickness (OR 0.52; 95% CI 0.320.85) and the distance between noncoronary cusp and the distal edge of the prosthesis (OR 1.37; 95% CI 1.031.83). Conclusions: Conduction abnormalities are frequently observed after TAVI with self-expandable bioprosthesis and definitive pacing is required in about a third of the patients, with a clear association with depth of implant and small interventricular septum thickness. (c) 2011 Wiley Periodicals, Inc.