Promising System for Selecting Healthy In Vitro–Fertilized Embryos in Cattle

Conventionally, in vitro–fertilized (IVF) bovine embryos are morphologically evaluated at the time of embryo transfer to select those that are likely to establish a pregnancy. This method is, however, subjective and results in unreliable selection. Here we describe a novel selection system for IVF bovine blastocysts for transfer that traces the development of individual embryos with time-lapse cinematography in our developed microwell culture dish and analyzes embryonic metabolism. The system can noninvasively identify prognostic factors that reflect not only blastocyst qualities detected with histological, cytogenetic, and molecular analysis but also viability after transfer. By assessing a combination of identified prognostic factors—(i) timing of the first cleavage; (ii) number of blastomeres at the end of the first cleavage; (iii) presence or absence of multiple fragments at the end of the first cleavage; (iv) number of blastomeres at the onset of lag-phase, which results in temporary developmental arrest during the fourth or fifth cell cycle; and (v) oxygen consumption at the blastocyst stage—pregnancy success could be accurately predicted (78.9%). The conventional method or individual prognostic factors could not accurately predict pregnancy. No newborn calves showed neonatal overgrowth or death. Our results demonstrate that these five predictors and our system could provide objective and reliable selection of healthy IVF bovine embryos

Low energy synchronous transcatheter cardioversion of atrial flutter/fibrillation in the dog

AbstractThe feasibility and effectiveness of low energy synchronous transcatheter cardioversion of atrial flutter and fibrillation were examined in dogs with talc-induced pericarditis, A conventional electrode catheter was positioned transvenously in the right atrial appendage. Atrial flutter/ fibrillation was induced by using the train pulse method, and the tachyarrhythmia-inducing threshold was determined. The minimal effective cardioversion energy levels were compared in three different cardioversion methods: method A = delivery of shock between the proximal electrode (cathode) and the backplate (anode), method B = delivery between the proximal electrode (cathode) and the distal electrode (anode) and method C = conventional external cardioversion.In both methods A and B, all 149 cardioversion attempts were successful with shocks of ≤5 J. Shocks of ≤1 J resulted in successful cardioversion in 57 (70%) of 81 attempts, 50 (74%) of 68 attempts and 5 (12%) of 41 attempts with methods A, B and C, respectively. The mean minimal effective cardioversion energy levels were not significantly different between methods A and B (0.62 ± 0.67 versus 0.58 ± 0.71 J). Transcatheter cardioversion decreased the defibrillation threshold 3- to 75-fold (men 6-to 7-fold) from that of transthoracic cardioversion. The defibrillation threshold was not influenced by the inducibility of atrial flutter/fibrillation. There were no complications of heart block, ventricular fibrillation or pathologic evidence of severe shock-induced atrial injury.Thus, low energy synchronous transcatheter cardioversion of atrial flutter/fibrillation is considered feasible and effective. This technique may also be useful in managing the atrial flutter/fibrillation that can occur during electrophysiologic studies

Effect of pre-in vitro maturation with cAMP modulators on the acquisition of oocyte developmental competence in cattle

The administration of follicle-stimulating hormone (FSH) prior to oocyte retrieval improves oocyte developmental competence. During bovine embryo production in vitro, however, oocytes are typically derived from FSH-unprimed animals. In the current study, we examined the effect of pre-in vitro maturation (IVM) with cAMP modulators, also known as the second messengers of FSH, on the developmental competence of oocytes derived from small antral follicles (2–4 mm) of FSH-unprimed animals. Pre-IVM with N6,2ʹ-O-dibutyryladenosine 3′,5′-cyclicmonophosphate (dbcAMP) and 3-isobutyl-1-methylxanthine (IBMX) for 2 h improved the blastocyst formation in oocytes stimulated by FSH or amphiregulin (AREG). Furthermore, pre-IVM enhanced the expression of the FSH- or AREG-stimulated extracellular matrix-related genes HAS2, TNFAIP6, and PTGS2, and epidermal growth factor (EGF)-like peptide-related genes AREG and EREG. Additionally, pre-IVM with dbcAMP and IBMX enhanced the expression of EGFR, and also increased and prolonged cumulus cell-oocyte gap junctional communication. The improved oocyte development observed using the pre-IVM protocol was ablated by an EGF receptor phosphorylation inhibitor. These results indicate that pre-IVM with cAMP modulators could contribute to the acquisition of developmental competence by bovine oocytes from small antral follicles through the modulation of EGF receptor signaling and oocyte-cumulus/cumulus-cumulus gap junctional communication

Transcriptomic signature of the follicular somatic compartment surrounding an oocyte with high developmental competence

During antral folliculogenesis, developmental competence of prospective oocytes is regulated in large part by the follicular somatic component to prepare the oocyte for the final stage of maturation and subsequent embryo development. The underlying molecular mechanisms are poorly understood. Oocytes reaching the advanced stage of follicular growth by administration of exogenous follicle-stimulating hormone (FSH) possess higher developmental competence than oocytes in FSH-untreated smaller follicles. In this study, the transcriptomic profile of the cumulus cells from cows receiving FSH administration (FSH-priming) was compared, as a model of high oocyte competence, with that from untreated donor cows (control). Ingenuity Pathway Analysis showed that cumulus cells receiving FSH-priming were rich in down-regulated transcripts associated with cell movement and migration, including the extracellular matrix-related transcripts, probably preventing the disruption of cell-to-cell contacts. Interestingly, the transcriptomic profile of up-regulated genes in the control group was similar to that of granulosa cells from atretic follicles. Interferon regulatory factor 7 was activated as the key upstream regulator of FSH-priming. Thus, acquisition of developmental competence by oocytes can be ensured by the integrity of cumulus cells involved in cell-to-cell communication and cell survival, which may help achieve enhanced oocyte-somatic cell coupling

The effect of the number of blastomeres at the onset of the lag-phase on cell numbers, ICM ratio, and apoptosis incidence.

<p>The number of total, ICM, and TE cells, the ICM ratio, and the apoptotic incidence in blastocysts with different numbers of blastomeres at the onset of the lag-phase are shown in <b>A</b> and <b>D</b>, respectively. Embryos were separated into groups based on high (<b>B</b> and <b>E</b>) or low oxygen consumption (<b>C</b> and <b>D</b>) with a cut-off of 0.84×10⁻¹⁴ mol s⁻¹. Data are presented as the mean ± SD. Different letters indicate significant differences between groups based on one-way ANOVA followed by Fisher's protected least signicicant difference (PLSD) (^A,B<i>P</i><0.01; ^a,b<i>P</i><0.05).</p