Gene expression profile in heat-shocked Holstein and Nelore oocytes and cumulus cells.

The present study determined the transcriptome profile in Nelore and Holstein oocytes subjected to heat shock during IVM and the mRNA abundance of selected candidate genes in Nelore and Holstein heat-shocked oocytes and cumulus cells (CC). Holstein and Nelore cows were subjected to in vivo follicle aspiration. Cumulus-oocyte complexes were assigned to control (38.5°C, 22h) or heat shock (41°C for 12h, followed by 38.5°C for 10h) treatment during IVM. Denuded oocytes were subjected to bovine microarray analysis. Transcriptome analysis demonstrated 127, nine and six genes were differentially expressed between breed, temperature and the breed×temperature interaction respectively. Selected differentially expressed genes were evaluated by real-time polymerase chain reaction in oocytes and respective CC. The molecular motor kinesin family member 3A (KIF3A) was upregulated in Holstein oocytes, whereas the pro-apoptotic gene death-associated protein (DAP) and the membrane trafficking gene DENN/MADD domain containing 3 (DENND3) were downregulated in Holstein oocytes. Nelore CC showed increased transcript abundance for tight junction claudin 11 (CLDN11), whereas Holstein CC showed increased transcript abundance for antioxidant metallothionein 1E (MT1E) . Moreover, heat shock downregulated antioxidant MT1E mRNA expression in CC. In conclusion, oocyte transcriptome analysis indicated a strong difference between breeds involving organisation and cell death. In CC, both breed and temperature affected mRNA abundance, involving cellular organisation and oxidative stress.

Machine Perfusion of Donor Livers for Transplantation: A Proposal for Standardized Nomenclature and Reporting Guidelines.

With increasing demand for donor organs for transplantation, machine perfusion (MP) promises to be a beneficial alternative preservation method for donor livers, particularly those considered to be of suboptimal quality, also known as extended criteria donor livers. Over the last decade, numerous studies researching MP of donor livers have been published and incredible advances have been made in both experimental and clinical research in this area. With numerous research groups working on MP, various techniques are being explored, often applying different nomenclature. The objective of this review is to catalog the differences observed in the nomenclature used in the current literature to denote various MP techniques and the manner in which methodology is reported. From this analysis, we propose a standardization of nomenclature on liver MP to maximize consistency and to enable reliable comparison and meta-analyses of studies. In addition, we propose a standardized set of guidelines for reporting the methodology of future studies on liver MP that will facilitate comparison as well as clinical implementation of liver MP procedures.

2016 Comprehensive Update of the Banff Working Group on Liver Allograft Pathology: Introduction of Antibody-Mediated Rejection.

The Banff Working Group on Liver Allograft Pathology reviewed and discussed literature evidence regarding antibody-mediated liver allograft rejection at the 11th (Paris, France, June 5-10, 2011), 12th (Comandatuba, Brazil, August 19-23, 2013), and 13th (Vancouver, British Columbia, Canada, October 5-10, 2015) meetings of the Banff Conference on Allograft Pathology. Discussion continued online. The primary goal was to introduce guidelines and consensus criteria for the diagnosis of liver allograft antibody-mediated rejection and provide a comprehensive update of all Banff Schema recommendations. Included are new recommendations for complement component 4d tissue staining and interpretation, staging liver allograft fibrosis, and findings related to immunosuppression minimization. In an effort to create a single reference document, previous unchanged criteria are also included.

Liver preservation with machine perfusion and a newly developed cell-free oxygen carrier solution under subnormothermic conditions.

We describe a new preservation modality combining machine perfusion (MP) at subnormothermic conditions(21 °C) with a new hemoglobin-based oxygen carrier (HBOC) solution. MP (n=6) was compared to cold static preservation (CSP; n=6) in porcine orthotopic liver transplants after 9 h of cold ischemia and 5-day follow-up. Recipients' peripheral blood, serial liver biopsies, preservation solutions and bile specimens were collected before, during and after liver preservation. Clinical laboratorial and histological analyses were performed in addition to mitochondrial functional assays, transcriptomic, metabolomic and inflammatory inflammatory mediator analyses. Compared with CSP, MP animals had: (1) significantly higher survival (100%vs. 33%; p<0.05); (2) superior graft function (p<0.05);(3) eight times higher hepatic O2 delivery than O2 consumption (0.78 mL O2/g/h vs. 0.096 mL O2/g/h) during MP; and (4) significantly greater bile production (MP=378.5 ± 179.7; CS=151.6 ± 116.85). MP downregulated interferon (IFN)-α and IFN-γ in liver tissue. MP allografts cleared lactate, produced urea, sustained gluconeogenesis and produced hydrophilic bile after reperfusion. Enhanced oxygenation under subnormothermic conditions triggers regenerative and cell protective responses resulting in improved allograft function. MP at 21 °C with the HBOC solution significantly improves liver preservation compared to CSP.

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

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 22 h in TCM-199 supplemented with 0, 2.5, 10 or 50 ng/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, 2.5 ng/ml FGF10 increased (p < 0.05) the percentage of oocytes with extrusion of the first polar body (35%) compared to 0, 10 and 50 ng/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 50 ng/ml FGF10 to increase the relative abundance of COX2 and PLAC8 and of 10 ng/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.

Mechanical properties of native and decellularized reproductive tissues: insights for tissue engineering strategies.

Understanding the mechanical properties and porosity of reproductive tissues is vital for regenerative medicine and tissue engineering. This study investigated the changes in Young's modulus (YM), storage modulus (E'), loss modulus (E"), and porosity of native and decellularized bovine reproductive tissues during the estrous cycle. Testis tunica albuginea had significantly higher YM, E', and E" than the inner testis, indicating greater stiffness and viscoelasticity. Endometrium showed no distinct differences in YM, E', or E" across the estrous cycle or between horns. Ovaries exhibited significant variations in YM, E', E", and porosity, with higher YM and E' in the ipsilateral cortex and medulla during the luteal phase. Decellularized ovarian tissues displayed increased porosity. The oviduct displayed no significant differences in YM or E' in the isthmus, but the contralateral ampulla had reduced YM and E' in the luteal phase. These findings offer valuable insights into the dynamic mechanical properties and porosity of reproductive tissues, facilitating the development of biomimetic scaffolds for tissue engineering applications.

Early treatment of depressive symptoms and long-term survival after liver transplantation.

While depression after liver transplantation (LTX) is associated with decreased survival, the effects of treating depression remain unknown. We assessed a previously described, prospective cohort of 167 patients transplanted for alcohol-related liver disease from 1998 to 2003. Depressive symptoms were measured with the Beck Depression Inventory serially throughout the first posttransplant year. Adequacy of antidepressant treatment was measured with the Antidepressant Treatment History Form. Using Cox-proportional Hazards modeling, survival times were assessed for recipients with no depression versus depression with adequate medications versus depression with inadequate medications. Seventy-two recipients had depressive symptoms in the first posttransplant year. Of these, 43% (n=31) received adequate pharmacotherapy and 57% (n=41) received inadequate (n=7) or no pharmacotherapy (n=34). After a median follow-up time of 9.5 years, 32% of the inadequately treated depressed group survived versus 52% of the adequately treated group and 56% of the nondepressed group (p=0.006). Compared to the nondepressed group, those with adequately treated depression had no significant difference in survival. However, recipients with depression and inadequate pharmacotherapy had decreased survival times compared to nondepressed recipients (HR for death=2.44, 95% CI=1.45, 4.11), controlling for other known confounders. The factor most strongly linked to long-term mortality after liver transplantation in this cohort was untreated depression.

Supplementing Ca salts of soybean oil via low-moisture molasses-based blocks to improve reproductive performance and overall productivity of beef cows.

This experiment evaluated reproductive and productive responses of beef cows receiving self-fed low-moisture blocks (LMB) enriched or not with Ca salts of soybean oil (CSSO) throughout the breeding season. Non-pregnant, suckled multiparous Angus-influenced cows were assigned to a fixed-time artificial insemination (AI) protocol (day -10 to 0) followed by natural service (day 15-70). Cows were managed in 12 groups (46 ± 4 cows/group) maintained in individual pastures, and groups received LMB enriched with 25 % (as-fed basis) of CSSO or ground corn (CON) from day - 10 to 100. Both treatments were designed to yield a daily LMB intake of 0.454 kg/cow (as-fed basis). Cows receiving CSSO had greater (P < 0.01) mean concentrations of ω-6 fatty acids in plasma samples collected on days 0 and 55. Cows receiving CSSO had greater (P = 0.05) pregnancy rate to fixed-time AI (67.2 vs. 59.3 %), whereas final pregnancy rate did not differ (P = 0.92) between treatments. Pregnancy loss was less (P = 0.03) in CSSO cows (4.50 vs. 9.04 %), which also calved earlier during the calving season (treatment × week; P = 0.04). Weaning rate tended to be greater (P = 0.09) in CSSO (84.8 vs. 79.4 %), although calf weaning age and weight did not differ (P ≥ 0.72) between treatments. Kilos of calf weaned/cow exposed was greater (P = 0.04) in CSSO cows (234 vs. 215 kg). Therefore, supplementing CSSO to beef cows via LMB during the breeding season improved their reproductive and overall productivity during a cow-calf cycle.

Re-examination of the lymphocytotoxic crossmatch in liver transplantation: can C4d stains help in monitoring?

C4d-assisted recognition of antibody-mediated rejection (AMR) in formalin-fixed paraffin-embedded tissues (FFPE) from donor-specific antibody-positive (DSA+) renal allograft recipients prompted study of DSA+ liver allograft recipients as measured by lymphocytotoxic crossmatch (XM) and/or Luminex. XM results did not influence patient or allograft survival, or cellular rejection rates, but XM+ recipients received significantly more prophylactic steroids. Endothelial C4d staining strongly correlates with XM+ (<3 weeks posttransplantation) and DSA+ status and cellular rejection, but not with worse Banff grading or treatment response. Diffuse C4d staining, XM+, DSA+ and ABO- incompatibility status, histopathology and clinical-serologic profile helped establish an isolated AMR diagnosis in 5 of 100 (5%) XM+ and one ABO-incompatible, recipients. C4d staining later after transplantation was associated with rejection and nonrejection-related causes of allograft dysfunction in DSA- and DSA+ recipients, some of whom had good outcomes without additional therapy. Liver allograft FFPE C4d staining: (a) can help classify liver allograft dysfunction; (b) substantiates antibody contribution to rejection; (c) probably represents nonalloantibody insults and/or complete absorption in DSA- recipients and (d) alone, is an imperfect AMR marker needing correlation with routine histopathology, clinical and serologic profiles. Further study in late biopsies and other tissue markers of liver AMR with simultaneous DSA measurements are needed.

Expression of bta-miR-222 and LHCGR in bovine cultured granulosa cells: Impact of follicle deviation and regulation by FSH/insulin in vitro.

Final antral follicle development and future ovulation are mediated by gonadotropin-induced changes with spatio-temporally regulated expression of genes. Here, we aimed to quantify the relative mRNA abundance of bta-miR-222 and its predicted target, LHCGR, in granulosa cells (GCs) from follicles, after follicle deviation, as well as from GCs cultured in vitro with follicle stimulating hormone (FSH) and/or insulin. Thus, to study the impact of follicle deviation, Nelore heifers (n = 10; Bos taurus indicus) were hormonally synchronized and slaughtered 3 days after ovulation. Then, GCs from the dominant follicle (DF) and its respective subordinate follicle (SF) were recovered for RT-qPCR. For in vitro analysis, small follicles (2-5 mm) were dissected from bovine ovaries collected from a local abattoir. The GCs were isolated and cultured in serum-free medium, or treated with insulin (1 ng/mL or 10 ng/mL) alone or in combination with human recombinant FSH (1 ng/mL), for 6 days. Our findings showed that the relative mRNA abundance of LHCGR in GCs was higher in the DF compared to the SF (p = 0.01). Inversely, bta-miR-222 expression was lower in the DF compared to the SF (p = 0.01). Furthermore, GCs cultured with FSH and insulin together resulted in a higher abundance of LHCGR and a lower abundance of bta-miR-222 (p ≤ 0.05) when compared to GCs cultured with insulin alone. In conclusion, we found that the LHCGR upregulation in GCs from the DF is inversely related to bta-miR-222 expression. We also suggest the involvement of FSH in bta-miR-222 suppression in healthy bovine GCs.

Early trajectories of depressive symptoms after liver transplantation for alcoholic liver disease predicts long-term survival.

Although it is well known that depression is associated with poorer medical outcomes, the association between depression- and liver transplant (LTX)-specific outcomes has not been investigated. We identified three trajectories of depressive symptoms evolving within the first post-LTX year in a cohort of 167 patients transplanted for alcoholic cirrhosis: a group with consistently low depression levels at all time points (group 1, n = 95), a group with initially low depression levels that rose over time (group 2, n = 41), and a group with consistently high depression levels (group 3, n = 31). Controlling for medical factors associated with poorer survival, recipients with increasing depression or persisting depression were more than twice as likely to die (all cause mortality) within the subsequent years. At 10 years post-LTX the survival rate was 66% for the low depression group, but only 46% and 43%, respectively, for the increasing depression and high depression groups. Except for a paradoxically higher percentage of malignancies in the low depression group, the causes of death and other specific LTX outcomes were not different between groups. Whether treatment of depression will improve survival rates is an area for research.

Effects of bismuth subsalicylate and calcium-ammonium nitrate on ruminal in vitro fermentation of bahiagrass hay with supplemental molasses.

There is a need to increase efficiency of beef production. Decreasing losses of CH4 and improving byproduct utilization are popular strategies. Two feed additives were tested to find potential solutions. Three randomized complete block design experiments were performed using batch culture systems to evaluate the effects of bismuth subsalicylate (BSS) and calcium-ammonium nitrate (CAN) on in vitro ruminal fermentation of bahiagrass hay and supplemental molasses. The first experiment contained four treatments: (1) basal substrate; (2) basal substrate with 0.75% urea (DM basis); (3) basal substrate with 1.2% CAN and 0.38% urea (DM basis); and (4) basal substrate with 2.4% CAN (DM basis). Treatments 2, 3, and 4 were isonitrogenous. The second experiment had a 4 × 3 factorial arrangement of treatments with 4 concentrations of BSS (0.00, 0.33, 0.66, and 1.00%; DM basis) and 3 concentrations of CAN (0.0, 1.2, and 2.4%; DM basis). The third experiment had the following treatments: (1) basal substrate; (2) basal substrate with 0.05% BSS (DM basis); (3) basal substrate with 0.10% BSS (DM basis); and (4) basal substrate with 0.33% BSS (DM basis). For all experiments, basal substrate consisted of Pensacola bahiagrass hay (Paspalum notatum Flüggé; 80% substrate DM) and molasses (20% substrate DM). All data were analyzed using the MIXED procedure of SAS. In Exp. 1, in vitro organic matter (OM) digestibility (IVOMD) was linearly reduced (P < 0.001) with the inclusion of CAN, and CH4, in mmol/g OM fermented, was decreased linearly (P < 0.001). The volatile fatty acid (VFA) profile was not impacted by the inclusion of nonprotein nitrogen (NPN) or CAN (P > 0.05). In Exp. 2, except for CH4 production (P < 0.05), there were no BSS × CAN interactions. Linear reductions in total gas production (P < 0.001), IVOMD (P < 0.001), and total concentration of VFA (P = 0.007) were observed with the inclusion of BSS up to 1%. The inclusion of BSS decreased H2S production in a quadratic manner (P = 0.024). In Exp. 3, IVOMD was not impacted by the inclusion of BSS (P > 0.05); however, production of H2S was linearly decreased (P = 0.004) with the inclusion of BSS up to 0.33%. In conclusion, in vitro fermentation was negatively impacted by the inclusions of BSS, up to 1%, and CAN, up to 2.4%; however, BSS decreased production of H2S when included up to 0.33% without impeding fermentation, while CAN decreased CH4 production.

Effects of nutrient restriction on the metabolic profile of Bos indicus-influenced and B. taurus suckled beef cows.

Recent research from our group demonstrated that Bos indicus-influenced suckled beef cows had greater resilience to withstand nutrient restriction and establish pregnancy compared with B. taurus cows exposed to the same conditions. To further understand these findings, differences in metabolic profile between these same B. indicus-influenced and B. taurus females were explored. Suckled beef cows (n = 134) were enrolled in a completely randomized design with a 2 × 2 factorial arrangement of treatments. On day -21, Angus (AN; Bos taurus) and Brangus (BN; B. indicus-influenced) cows were randomly assigned to 1) a diet that met daily energy maintenance requirements (MAINT), or 2) a diet that restricted intake to 70% of the daily energy maintenance requirements (RESTR). Cows were exposed to an estrus synchronization protocol and received an embryo 7 d after ovulation was pharmacologically induced on day 0. Blood samples were collected on days -21 and 19 to determine circulating concentrations of non-esterified fatty acids (NEFA), ß-hydroxybutyrate (BHB), insulin, glucose, and IGF-1. Pregnancy status after embryo transfer was determined on day 28. As a consequence of the proposed diets, cows in the RESTR diet had less body condition score (BCS) on day 19 (P = 0.008) across breed types. Moreover, BCS change from day -21 to 19 was included as independent covariate into subsequent analyses, allowing for the comparison of breed types under an equivalent level of body reserve mobilization. A breed × diet interaction was observed for plasma insulin (P = 0.03) and IGF-1 (P = 0.04) on day 19, where AN-RESTR cows had less plasma concentrations on day 19 compared with AN-MAINT cows. Diets did not impact (P > 0.10) plasma insulin and IGF-1 concentrations in BN cows. No diet or breed effects were observed in circulating concentrations of NEFA, BHB, and glucose (P > 0.10). Across breed types and nutritional treatment, there was positive linear effect (P ≤ 0.04) of plasma concentrations of insulin and IGF-1 on the probability of pregnancy to fixed-time embryo transfer. In summary, the negative impacts of nutrient restriction on the somatotropic axis, independently of body tissue mobilization, were heightened in Bos taurus females compared with B. indicus-influenced cohorts, which corroborate with the differences observed in fertility between these subspecies.

Pre-synchronization of ovulation timing and delayed fixed-time artificial insemination increases pregnancy rates when sex-sorted semen is used for insemination of heifers.

This study was conducted to determine effects of pre-synchronization of ovulation timing among heifers and delayed fixed-time artificial insemination (TAI) with sex-sorted semen on proportion of heifers pregnant after TAI (PR/AI). Heifers were assigned to one of eight treatments: 1 and 2), 7-d CO-Synch + CIDR treatment regimen with administration of gonadotropin-releasing hormone and a CIDR insert on Day 0, prostaglandin F2α (PGF) at CIDR removal on Day 7, and TAI occurring 54 h later with conventionally processed (CTRL54-CNV) or sex-sorted semen (CTRL54-SEX); 3 and 4), same as CTRL54 but TAI delayed to 72 h with conventionally processed (CTRL72-CNV) or sex-sorted semen (CTRL72-SEX); 5 and 6), same as CTRL54 but additional administration of PGF on Day -7 and TAI with conventionally processed (PRE54-CNV) or sex-sorted semen (PRE54-SEX); 7 and 8), same as PRE54 treatments but TAI delayed to 72 h with conventionally processed (PRE72-CNV) or sex-sorted semen (PRE72-SEX). Proportion of heifers pregnant after TAI was greater (P ≤  0.02) with conventionally processed semen compared with sex-sorted semen, yet PR/AI did not differ (P =  0.14) between heifers in PRE72-CNV and PRE72-SEX groups. There were greater PR/AI in the PRE72-SEX (P =  0.03) than CTRL54-SEX group (46.1 % and 36.9 %) and there was no difference (P =  0.31) in PR/AI between CTRL54-CNV and PRE72-SEX groups (50.4 % and 46.1 %). In conclusion, pre-synchronization of ovulation timing among heifers combined with delayed TAI resulted in increased PR/AI with sex-sorted semen compared with the 7-d CO-Synch+CIDR treatment regimen.

Trajectories of alcohol consumption following liver transplantation.

Any use of alcohol in the years following liver transplantation (LTX) approaches 50% of patients transplanted for alcoholic liver disease (ALD). We collected detailed prospective data on alcohol consumption following LTX for ALD to investigate ongoing patterns of use. Using trajectory modeling we identified four distinct alcohol use trajectories. One group had minimal use over time. Two other groups developed early onset moderate-to-heavy consumption and one group developed late onset moderate use. These trajectories demonstrate that alcohol use varies based on timing of onset, quantity and duration. Using discriminant function analysis, we examine characteristics of recipient's pre-LTX alcohol histories and early post-LTX psychological stressors to identify the profile of those at risk for these specific trajectories. We discuss the relevance of these findings to clinical care and preliminarily to outcomes.

Effects of ambient temperature and soybean meal supplementation on intake and digestion of two sheep breeds differing in mature size.

The aim of this study was to compare the intake and digestive physiology of mature ewes of two breeds--Ile-de-France (mature weight: 75-80 kg) and Churra-da-Terra-Quente (CTQ; mature weight: 45-50 kg)--and evaluate the effects of ambient temperature and protein supplementation in the comparison. The temperature (25 °C vs. 11 °C) and soybean meal supplementation (150 g/kg of ingested hay on dry matter basis vs. unsupplemented control) were evaluated in 48 adult ewes of two breeds fed hay ad libitum and at a restricted level of intake. The intake, digestibility, rumen pH and NH(3)-N, rumen outflow rates, faeces particle size and thyroid hormones levels were measured. These hormones can be related with gastrointestinal motility, thus explaining rumen outflow rate patterns. Dry matter intake per kg of body weight was higher in CTQ ewes (p < 0.05). This breed also exhibited lower organic matter and neutral detergent fibre digestibility (p < 0.001) and higher solid (p < 0.001) and liquid (p < 0.01) rumen outflow rates irrespective of intake level, supplementation or temperature. Rumen pH remained above 6.6 in all treatments. NH(3)-N rumen content was similar (p > 0.05) when breeds were fed only hay. There was no breed effect (p > 0.05) on faeces particle size. Triiodothyronine was not affected (p > 0.05) by breed and thyroxine was higher (p < 0.10) in the CTQ breed but only at the lower temperatures (breed × temperature, p < 0.05). Ile-de-France sheep showed a lack of adaptation to lower temperatures. This study suggests that the native CTQ breed fulfils its metabolic needs by having a higher intake and inherits faster flow through the gastrointestinal tract, as a result, its digestive ability is diminished.

Presynchronization with prostaglandin F2α and prolonged exposure to exogenous progesterone impacts estrus expression and fertility in beef heifers.

To determine the effects of two presynchronization strategies in conjunction with delayed fixed-time artificial insemination (TAI) on pregnancy rates to TAI (PR/AI), 1700 Angus beef heifers at three locations in South Dakota were enrolled in a completely randomized design with a 2 by 2 factorial arrangement of treatments. Within location, all heifers were randomly assigned to one of four treatments: 1) PG54 (n = 434), heifers were administered prostaglandin F2α (PGF; 25 mg im) 7 d prior [Day -14] to the initiation of the 7-d CO-Synch + controlled internal drug releasing (CIDR) protocol wherein they received gonadotropin-releasing hormone (GnRH; 100 µg im) and a CIDR insert on Day -7, PGF at CIDR removal on Day 0, and a second injection of GnRH concurrently with TAI 54 ± 2 h later; 2) PG72 (n = 426), heifers were exposed to the same treatment as PG54, however, TAI was performed 72 ± 2 h after CIDR removal; 3) PG-CIDR54 (n = 422), same as PG54 but heifers received a CIDR insert on Day -14 rather than Day -7, in addition to PGF administration; 4) PG-CIDR72 (n = 418), same as PG-CIDR54, however, TAI was performed 72 ± 2 h after CIDR removal. Estrus detection patches were applied to all heifers on Day 0 and were evaluated for activation at TAI. Pregnancy was diagnosed via transrectal ultrasonography between 30 and 47 d after TAI. The percentage of heifers exhibiting estrus between Day 0 and TAI was greater (P < 0.01) in the PG72, PG-CIDR54, and PG-CIDR72 treatments compared to the PG54 treatment (78.11, 86.59, and 91.09 vs. 31.05%, respectively). Furthermore, estrus response was greater (P < 0.01) in PG-CIDR72 heifers when compared to PG72. Pregnancy rates to TAI differed among treatments and were greater (P < 0.05) in the PG72 and PG-CIDR54 treatments when compared to PG-CIDR72 (48.8 and 50.4 vs. 38.4%, respectively), and were greater (P = 0.03) in PG-CIDR54 vs. PG54 (43.1%). Moreover, a tendency (P = 0.10) was determined on PR/AI between PG54 and PG72. In conclusion, presynchronization strategies and prolonged exposure to exogenous progesterone have the potential to alter estrus expression and improve fertility in replacement beef heifers.

Molecular and cellular effects of insulin-like growth factor-1 and LongR3-IGF-1 on in vitro maturation of bovine oocytes: comparative study.

Addition effects of insulin-like growth factor-1 (IGF-1) and its synthetic analogue insulin-like growth factor-1 recombinant-3 (LongR3-IGF-1) after in vitro maturation (IVM) of cattle cumulus-oocyte complexes (COCs) were compared and evaluated on meiotic progression, apoptosis and profile genes of oocyte competence (GDF9, BMP15, BAX, BCL2, OOSP1, IGFBP2, IGBFP4 and IGFBP5), and their respective cumulus cells (AREG, EGFR, FSHR, COX2, BAX, BCL2, IGFBP2, IGFBP4 and IGFBP5). The 739 COCs (n = 10 pools) of bovine ovaries were collected, selected and matured with IGF-1 (100 ng/mL), LongR3-IGF-1 (100 ng/mL), and in two control groups with 0.1% polyvinyl alcohol (PVA) or 10% fetal bovine serum (FBS), for 22-24 h. The statistical analysis was performed by a linear mixed effects model, ANOVA and Tukey tests. There was no statistical difference between experimental groups taken into account the meiotic progression and apoptosis (P > 0.05). Nevertheless, there were statistical differences (P ≤ 0.05) among FBS, IGF-1 and LongR3-IGF-1 groups for IGFBP4 gene expression, and among PVA, IGF-1 and LongR3-IGF-1 for COX2 gene expression in cumulus cells. Moreover, statistical difference was found for BCL2 gene expression between IGF-1, FBS and PVA groups and for IGFBP4 gene expression between LongR3-IGF-1, PVA and FBS in oocytes. There was no statistical difference between experimental groups for other genes evaluated. These results showed a good performance of IVM of bovine oocytes in the presence of LongR3-IGF-1 and the possibility of replacement of IGF-1 and FBS.