Inferences of body energy reserves on conception rate of suckled Zebu beef cows subjected to timed artificial insemination followed by natural mating

The influence of body condition score (BCS), rump fat thickness (RFAT), and live weight (LW), and the changes in these parameters during the interval from 165 of prepartum (i.e., 125 days of prior gestation) to 112 postpartum on first service conception and pregnancy rates were investigated in suckled Zebu (Bos indicus) beef cows (n = 266) subjected to timed artificial insemination (TAI) followed by natural mating. The aforementioned parameters were recorded at 165 ± 14 days (mean ± standard error) prepartum (concurrent with the weaning of previous calf), at parturition, and at 42 ± 7 days (at the onset of the synchronization of ovulation protocol), 82 ± 7 days (30 days after TAI), and 112 ± 7 days (60 days after TAI) postpartum. At the start of the breeding season (BS), cows were subjected to a synchronization of ovulation program for TAI. Bulls were placed with cows 10 days after TAI and remained until the end of the study (112 days postpartum). Cows with the highest BCS at parturition had an increased probability of first service conception rate at 60 days after TAI (P = 0.02) and a reduced probability of occurrence of pregnancy loss (P = 0.05). Also, cows had a greater likelihood of conceiving postpartum if they had greater RFAT and BCS at 165 ± 14 days prepartum (P = 0.01 and P = 0.03, respectively) and at parturition (P = 0.0007 and P = 0.003, respectively). Cows that had an increase in RFAT and BCS during the dry period (i.e., interval from weaning of the previous calf to parturition) also had a greater likelihood of conceiving (P = 0.03 and P = 0.06, respectively) during the BS. Among the different time points, RFAT and BCS at parturition had the largest impact on risk of conception during the BS. The LW was a poor predictor of conception during the BS (P = 0.11–0.68) except for LW at 165 ± 14 days prepartum (P = 0.01). Collectively, the findings indicated that the likelihood of conception during the BS was highest in cows that had an improvement in RFAT and BCS during the dry period. Therefore, assuring a good nutritional status in the dry period (BCS ≥ 3.0 at 165 ± 14 days prepartum and ≥3.25 at parturition) is an important aim to optimize the postpartum conception rate of suckled Zebu beef cows subjected to TAI followed by natural mating.     

Effect of interval from induction of puberty to initiation of a timed AI protocol on pregnancy rate in Nellore heifers

Prepubertal Bos indicus heifers (n = 774) were submitted to an E₂/P₄-based timed artificial insemination (TAI) protocol at three different intervals after induction of their pubertal ovulation by insertion of an intravaginal progesterone (P₄) device for 12 days. Heifers were randomly assigned to start the TAI protocol at 10 (group 10; n = 253), 12 (group 12; n = 265), or 14 (group 14; n = 256) days after the P₄ device was removed. The TAI protocol consisted of the following: insertion of intravaginal device containing P₄ (Controlled internal drug release [CIDR]; previously used twice for 9 days each) + estradiol benzoate (2 mg) on Day 0, CIDR withdrawal + estradiol cypionate (0.5 mg) and PGF2α (12.5 mg) on Day 9, and TAI on Day 11. A subgroup of heifers (n = 472) was evaluated by ultrasound on Days 9 and 11 to evaluate the ovaries and to determine P₄ concentrations on Day 9. On Day 9, more (P < 0.05) CLs were present, and follicular diameter was smaller (P < 0.05) for group 10 than for groups 12 and 14 (38.4%, 29.3%, and 23.3% with CL and 9.4 ± 0.1, 9.9 ± 0.1, and 9.8 ± 0.1 mm diameter, respectively), but P₄ concentrations did not differ (P > 0.1) between treatments (2.4 ± 0.06 ng/mL). Follicular diameter at TAI (11.08 ± 0.09 mm) and ovulation rate (88.4%) did not differ between treatments (P > 0.1). However, conception and pregnancy rates for all heifers were greater (P < 0.05) in group 12 (50.4% and 45.5%, respectively) than in group 10 (38.2% and 33.7%, respectively), with group 14 intermediate to other treatments (45.6% and 40.6%, respectively). The final pregnancy rate did not differ between treatments (80.9%). In conclusion, a 12-day interval from the end of the puberty induction protocol to the start of the TAI protocol resulted in greater conception and pregnancy rates in prepubertal Nellore heifers.     

Effects of rumen-protected methionine and choline supplementation on the preimplantation embryo in Holstein cows

Our objective was to determine the effects of supplementing methionine and choline during the prepartum and postpartum periods on preimplantation embryos of Holstein cows. Multiparous cows were assigned in a randomized complete-block design into four treatments from 21 days before calving to 30 days in milk (DIM). Treatments (TRT) were MET (n = 9, fed the basal diet + rumen-protected methionine at a rate of 0.08% [w:w] of the dry matter [DM], Smartamine M), CHO (n = 8, fed the basal diet + choline 60 g/d, Reashure), MIX (n = 11, fed the basal diet + Smartamine M and 60 g/d Reashure), and CON (n = 8, no supplementation, fed the close-up and fresh cow diets). Cows were randomly reassigned to two new groups (GRP) to receive the following diets from 31 to 72 DIM; control (CNT, n = 16, fed a basal diet) and SMT (n = 20, fed the basal diet + 0.08% [w:w] of the dry matter intake as methionine). An progesterone intravaginal insert (CIDR) device was inserted in all cows after follicular aspiration (60 DIM) and superovulation began at Day 61.5 using FSH in eight decreasing doses at 12-hour intervals over a 4-day period. On Days 63 and 64, all cows received two injections of PGF2α, and CIDR was removed on Day 65. Twenty-four hours after CIDR removal, ovulation was induced with GnRH. Cows received artificial insemination at 12 hours and 24 hours after GnRH. Embryos were flushed 6.5 days after artificial insemination. Global methylation of the embryos was assessed by immunofluorescent labeling of 5-methylcytosine, whereas lipid content was assessed by staining with Nile red. Nuclear staining was used to count the total number of cells per embryo. There was no difference between TRT, GRP, or their interaction (P > 0.05) for embryo recovery, embryos recovered, embryo quality, embryo stage, or cells per embryo. Methylation of the DNA had a TRT by GRP interaction (P = 0.01). Embryos from cows in CON-CNT had greater (P = 0.04) methylation (0.87 ± 0.09 arbitrary units [AU]) than embryos from cows in MET-CNT (0.44 ± 0.07 AU). The cytoplasmic lipid content was not affected (P > 0.05) by TRT or their interaction, but lipid content was greater (P = 0.04) for SMT (7.02 ± 1.03 AU) than that in CNT (3.61 ± 1.20 AU). In conclusion, cows in MET-CNT had embryos with lower methylation, and SMT cows had a higher lipid content than CNT. Methionine supplementation seems to impact the preimplantation embryo in a way that enhances its capacity for survival because there is strong evidence that endogenous lipid reserves serve as an energy substrate.     

Effects of Vascular Endothelial Growth Factor 165 on Bone Tissue Engineering

To study the relationship between vascular endothelial growth factor (VEGF) and formation and repair of engineering bone, second-generation bone marrow stromal cells (BMSCs) of New Zealand white rabbits that were separated in vitro were transfected with VEGF 165 gene vectors by adenovirus to detect gene expressions. Transfected BMSCs and β-tricalcium phosphate material were complexed and implanted at the femoral injury sites of the study group (n = 12), and the control group (n = 12) were implanted with engineering bones that were not transfected with VEGF. Femoral recoveries of the two groups were observed on the 15th, 30th, 45th and 60th days, and their vascularization and ossification statuses were observed by immunohistochemical methods. The BMSCs transfected with VEGF highly expressed VEGF genes and excreted VEGF. The two groups both experienced increased vascularization and bone volume after implantation (t = 7.92, P<0.05), and the increases of the study group were significantly higher than those of the control group (t = 6.92, P<0.05). VEGF is clinically applicable because it can accelerate the formation and repair of engineering bone by promoting vascularization and ossification.     

Circulating progesterone dynamics after intravaginal instillation of prostaglandin-F2α to lactating dairy cows

Our objectives were to evaluate circulating progesterone (P4) concentration dynamics and test the feasibility of inducing luteal regression after intravaginal (IVG) instillation of the PGF2α analogue dinoprost (PGF) in lactating dairy cows. In two experiments, cows were synchronized using the Ovsynch protocol to induce the formation of a corpus luteum (CL). Cows with at least one functional (P4 ≥1 ng/mL) CL ≥15 mm 7.5 days after Ovsynch remained in the studies. In experiment 1, cows (n = 31) were stratified by parity group and received 5 mL of saline IVG (SAL-IVG, n = 6), 25 mg of PGF intramuscular (IM) (PGF25-IM, n = 7), 25 mg of PGF IVG (PGF25-IVG, n = 6), 50 mg of PGF IVG (PGF50-IVG, n = 6), and 125 mg of PGF IVG (PGF125-IVG, n = 6). Experiment 2 was conducted to test the hypothesis that IVG instillation of two 25 mg doses of PGF 12 hours apart would be more effective than a 25- or 50-mg dose in a single application. Cows (n = 32) were stratified by parity and received SAL-IVG (n = 7), PGF25-IM (n = 7), PGF25-IVG (n = 6), and PGF50-IVG (n = 6) as in experiment 1, whereas another group received two IVG instillations of 25 mg of PGF 12 hours apart (PGF25-2X-IVG, n = 6). Blood was collected at −1 hour, every 6 hours from 0 hour to 24 hours, and every 12 hours up to 96 hours after treatment (trt). In experiment 1, there was an effect of trt (P < 0.01), time (P < 0.001), and an interaction between trt and time on P4 concentrations (P < 0.001). All PGF-treated groups had lower (P < 0.05) concentrations of P4 than cows in the SAL-IVG group from 12 to 96 hours after trt. Although an initial decline in P4 concentrations was induced in all PGF-treated cows, some cows in the IVG-treated groups presented a rebound in plasma P4, indicating CL recovery. More cows in the PGF25-IVG and PGF125-IVG groups than in the PGF50-IVG and PGF25-IM groups presented CL recovery, suggesting that greater doses of PGF may not necessarily improve CL regression. In experiment 2, there was an effect of trt (P < 0.001), time (P < 0.001), and an interaction between trt and time on P4 concentrations (P < 0.001). All PGF-treated groups had lower (P < 0.05) P4 than the SAL-IVG group from 12 to 96 hours after trt. Cows in the PGF25-2X-IVG group had a P4 profile that was similar to that of cows in the PGF25-IM group and the lowest P4 concentrations after treatment among the IVG-treated groups, and all cows presented complete CL regression (defined as P4 <0.4 ng/mL). We conclude that CL regression can be induced through IVG instillation of PGF in lactating dairy cows and that instillation of two IVG doses of 25 mg of PGF 12 hours apart was the most effective strategy.     

Pluripotent stem cell-derived mesenchymal stromal cells improve cardiac function and vascularity after myocardial infarction

Background aimsMesenchymal stromal cells (MSCs) have been shown to improve cardiac function after injury and are the subject of ongoing clinical trials. In this study, the authors tested the cardiac regenerative potential of an induced pluripotent stem cell-derived MSC (iPSC-MSC) population (Cymerus MSCs) in a rat model of myocardial ischemia-reperfusion (I/R). Furthermore, the authors compared this efficacy with bone marrow-derived MSCs (BM-MSCs), which are the predominant cell type in clinical trials.MethodsFour days after myocardial I/R injury, rats were randomly assigned to (i) a Cymerus MSC group (n = 15), (ii) a BM-MSC group (n = 15) or (iii) a vehicle control group (n = 14). For cell-treated animals, a total of 5 × 10⁶ cells were injected at three sites within the infarcted left ventricular (LV) wall.ResultsOne month after cell transplantation, Cymerus MSCs improved LV function (assessed by echocardiography) compared with vehicle and BM-MSCs. Interestingly, Cymerus MSCs enhanced angiogenesis without sustained engraftment or significant impact on infarct scar size. Suggesting safety, Cymerus MSCs had no effect on inducible tachycardia or the ventricular scar heterogeneity that provides a substrate for cardiac re-entrant circuits.ConclusionsThe authors here demonstrate that intra-myocardial administration of iPSC-MSCs (Cymerus MSCs) provide better therapeutic effects compared with conventional BM-MSCs in a rodent model of myocardial I/R. Because of its manufacturing scalability, iPSC-MSC therapy offers an exciting opportunity for an “off-the-shelf” stem cell therapy for cardiac repair.     

Effect of PGF2α and GnRH on the reproductive performance of postpartum dairy cows subjected to synchronization of ovulation and timed artificial insemination during the warm or cold periods of the year

This study was designed to evaluate the reproductive performance of lactating dairy cows (Holstein Friesian) after the injection of PGF_2α analogue on Day 15 postpartum, and GnRH analogue on Day 23 after artificial insemination (AI) with Presynch (two injections of PGF_2α, administered 14 days apart starting at 30–35 days postpartum) + Ovsynch-based (GnRH–7 days–PGF_2α–2 days–GnRH–16–20 hours–timed artificial insemination) treatments, during the warm and cold periods of the year. All the cows (n = 313) were assigned to one of the four groups including: M₁ (n = 72) in which the cows were treated with PGF_2α on Day 15 postpartum + Presynch-Ovsynch + GnRH on Day 23 post-AI; M₂ (n = 41) in which the cows received PGF_2α on Day 15 postpartum + Presynch-Ovsynch; M₃ (n = 100) including the cows that got Presynch-Ovsynch; and control group (n = 100) including the cows that were not treated and were inseminated at natural estrus. Pregnancy diagnosis was performed 28 to 35 days post-insemination by means of ultrasound. The results showed that treatment with PGF_2α on Day 15 postpartum significantly decreased the days to conception and the number of services per conception (P < 0.01) and it also improved the first service conception rate (P < 0.1) only in cows that were treated with M₂ protocol. Whereas, the days to first service was not influenced by the treatment of PGF_2α on Day 15 postpartum (P > 0.05). In contrast, administration of GnRH on Day 23 post-AI increased the days to conception and the number of service per conception (P < 0.01) and tended to decrease the first service conception rate (P < 0.1) in cows that were treated with M₁ compared with M₂ protocol. Therefore, it was concluded that Presynch-Ovsynch protocol could be more reproductive and beneficial when a single treatment with PGF_2α was administered at 15 days postpartum (15 days after the PGF_2α, Presynch-Ovsynch protocol was initiated). Interestingly, the administration of a GnRH agonist on Day 23 post-AI not only did not improve the reproductive performance of the cows receiving first postpartum timed artificial insemination after Presynch-Ovsynch protocol but also reduced that.     

Effects of dietary supplementation of pioglitazone on metabolism,milk yield,and reproductive performance in transition dairy cows

The objective of this study was to investigate the effect of dietary supplementation of pioglitazone (PGT), a specific ligand for PPARγ, on metabolic dynamics, milk production, and reproductive performance of transition dairy cows. Eighty multiparous Holstein cows in their second or more lactations were blocked by the calving date and parity and assigned randomly to four dietary groups (n = 20 cow/treatment) including control (no PGT−/−), supplemented with PGT (6-mg PGT/kg body weight) from Day −14 to +21 relative to parturition (PGT+/+) or only during prepartum (PGT+/−) or postpartum periods (PGT−/+). Postpartum body condition score and body weight loss decreased (P < 0.05) in all PGT-supplemented groups. Milk yield was not affected by PGT supplementation (P > 0.05). Percentage of milk fat decreased (P < 0.05) in all PGT-treated groups; however, milk fat yield was lower (P < 0.05) in PGT (+/+) and PGT (+/−) groups compared with PGT (−/−). Peripartum (Day −7 to +7) concentrations of plasma nonesterified fatty acids and β-Hydroxybutyrate decreased in PGT (+/+) but not in the PGT (−/−) group (P < 0.05). During the postpartum period, PGT reduced (P > 0.05) plasma concentrations of nonesterified fatty acids in all PGT-treated groups but did not affect β-Hydroxybutyrate level. Plasma concentrations of triglycerides decreased in all PGT-supplemented groups. Supplementation of PGT increased the peripartum concentrations of plasma glucose in PGT (+/+) and PGT (+/−) groups compared with control. Plasma concentrations of insulin-like growth factor 1 were higher in PGT (+/+) compared with the control group during both the peripartum and postpartum periods. Plasma concentrations of growth hormone and insulin were not affected by PGT treatment (P > 0.05). Mean days to ovulation were lower in PGT (+/+) and PGT (-/+), and the proportion of cows ovulating by Day 14 postpartum was higher in PGT (+/+) compared with control. Days open were shorter in PGT (+/+), PGT (+/−), and PGT (−/+) groups compared with control. However, the proportion of pregnant cows at 120 days in milk were higher in all PGT-supplemented groups. The results showed positive effects of dietary supplementation of PGT, especially supplementation during both the prepartum and postpartum periods, on metabolic dynamics, ovarian function, and reproductive performance in transition dairy cows.     

Ovarian follicular growth suppression by long-term treatment with a GnRH agonist and impact on small follicle number,oocyte yield,and in vitro embryo production in Zebu beef cows

The aim of the present study was to evaluate small follicle number, oocyte yield, and in vitro embryo production (IVEP) in Zebu beef cows treated long term with a GnRH agonist to suppress ovarian follicular growth. Nelore (Bos indicus) cows (n = 20) showing regular estrous cycles were randomly assigned to one of two groups: control (n = 10, placebo ear implant without a GnRH agonist); GnRH agonist (n = 10, GnRH agonist ear implant containing 9.4-mg deslorelin). All cows underwent an ovum pick-up (OPU) session 14 days (Day 14) before the start of treatments (Day 0) followed by seven OPU–IVEP procedures at 30-day intervals (Days 0, 30, 60, 90, 120, 150, and 180). Semen from a single batch of a previously tested bull was used for all the IVEP. Cows treated with agonist reported a decrease over time in the proportion of animals with a (CL; P ≤ 0.05) and large follicles (>10 mm, P ≤ 0.05). These cows had a lesser number of medium + large follicles (>5 mm; 1.74 ± 0.5 vs. 4.13 ± 0.5; P ≤ 0.05), greater number of small follicles (2–5 mm; 44.3 ± 2.8 vs. 30.8 ± 1.8; P ≤ 0.05), greater yield of cumulus-oocyte complexes (COCs; 21.0 ± 2.3 vs. 15.6 ± 1.9; P ≤ 0.05), greater proportion of COCs cultured (79.2 vs. 73.9%; P ≤ 0.05), COCs cleaved (10.6 ± 1.5 vs. 6.8 ± 1.1, P ≤ 0.05), and cleaved rate (52.8 vs. 44.3%; P ≤ 0.05) compared with control cows. The number (3.4 ± 0.7 vs. 3.0 ± 0.6; P > 0.05) and proportion (16.5 vs. 19.1%; P > 0.05) of blastocysts produced were similar between agonist and control cows, respectively. The study has shown that Zebu beef cows treated long term with a GnRH agonist had follicular growth restricted to small follicles. This did not compromise the ability of oocytes to undergo IVF and embryonic development.     

Effects of exogenous progesterone and cloprostenol on ovarian follicular development and first ovulation in prepubertal heifers

The objective of this study was to determine the effects of progesterone and cloprostenol (a PGF_2α analogue) on ovarian follicular development and ovulation in prepubertal heifers. In Experiment 1, crossbred Hereford heifers (Bos taurus; 10 to 12 mo old, 255 to 320 kg) were assigned randomly to three groups and given (1) an intravaginal progesterone-releasing insert (CIDR; P group, n = 13); (2) a CIDR plus 500 μg cloprostenol im (PGF_2α analogue) at CIDR removal (PPG group, n = 11); or (3) no treatment (control group, n = 14). The CIDR inserts were removed 5 d after follicular wave emergence. Progesterone-treated heifers (P and PPG groups) had a larger dominant follicle than that of the control group (P = 0.01). The percentage ovulating was highest in the PPG group (8 of 11, 73%), intermediate in the P group (4 of 13, 31%), and lowest in the control group (1 of 14, 7%; P < 0.02). In Experiment 2, 16 heifers (14 to 16 mo old, 300 to 330 kg) were designated to have follicular wave emergence synchronized with either a CIDR and 1 mg estradiol benzoate im (EP group, n = 8) on Day 0 (beginning of experiment) or by transvaginal ultrasound-guided ablation of all follicles ≥5 mm on Day 3 (FA group, n = 8). On Day 7, CIDRs were removed in the EP group, and all heifers received 500 μg cloprostenol im. Ovulation was detected in 6 of 8 heifers (75%) in both groups. In summary, the use of PGF_2α with or without exogenous progesterone treatment increased the percentage ovulating in heifers close to spontaneous puberty.     

Therapeutic effect of autologous bone marrow stem cell mobilization combined with anti-infective therapy on moyamoya disease

ObjectiveThe purpose of this study is to explore the therapeutic effect of autologous bone marrow stem cell (ABMSC) mobilization combined anti-infection therapy on patients with moyamoya disease (MMD), and to provide reference for the clinical treatment of MMD and cerebrovascular disease.Methods54 adult patients with MMD diagnosed in Henan Provincial People’s Hospital from March 2017 to March 2019 were chosen as research objects. All patients were randomly divided into study group (SG) and control group (CG), with 27 patients in each group. Patients in both groups received conventional drug treatment after diagnosis of MMD, and received dura turnover of brain - temporal muscle - superficial temporal artery application surgery during indirect vascular reconstruction. On the basis of surgical treatment, patients in the SG were given ABMSC mobilization combined with low-dose dexamethasone for anti-inflammatory and anti-infection treatment. ABMSCs were mobilized by recombinant human granulocyte colony stimulating factor (rhG-csF) and recombinant human granulocyte - macrophage colony stimulating factor (rhoM-esF). The therapeutic effects of the patients were evaluated BF, one month after treatment (AF), three months AF, and six months AF. The number of hematopoietic stem cells (HpCs) and inflammatory indicators were compared between the two groups before and 4 weeks AF.ResultsFirstly, the Barthcl index of patients in the two groups showed a gradual increase trend at the 3rd and 6th months AF, and the ascensional range in the research group was higher than that in the CG (P < 0.05). Secondly, at the 3rd and 6th month AF, national institute of heath stroke scale (NIHSS) scores of patients in the CG were lower than those before treatment (BF), and there was an important change in NIHSS scores between the two groups at the same period (P < 0.05). Thirdly, after 1 month of treatment and 3 months of treatment, Chinese stroke scale (CSS) scores of patients in both groups decreased obviously compared with those BF, and the SG was lower than the CG, with statistical changes (P < 0.05). Fourthly, after 4 weeks of treatment, the hematopoietic stem cell counts in both groups were higher than those BF, and the hematopoietic stem cell counts in the SG were obviously higher than those in the CG (P < 0.05). All three inflammatory indicators were improved compared with those BF, and the SG was better than the CG (P < 0.05).ConclusionAutogenous bone marrow stem cell mobilization combined with dexamethasone anti-inflammation and anti-infection treatment after revascularization in patients with MMD can accelerate the recovery of nerve function and promote the formation of new blood vessels. At the same time, it can reduce inflammation and improve patients' quality of life, which is worthy of clinical reference.     

Ameliorative effects of L-glutamine on haematological parameters in heat-stressed Red Sokoto goats

The aim of the study was to evaluate haematological responses in Red Sokoto goats (RSGs) administered with L-glutamine during the hot-dry season. Experimental animals included 28 clinically healthy RSGs divided into treated group (n = 14); each administered L-glutamine at 0.2 g/kg body weight, dissolved in 10 mL distilled water, and control group (n = 14); each administered 10 mL distilled water, per os once daily for 21 days. The ambient temperature and relative humidity recorded daily for 4 weeks were used to calculate the temperature-humidity index. Three millilitres of blood sample was collected from each goat by jugular venipuncture for haematology, while rectal temperature (RT), heart rate (HR) and respiratory rate (RR) were also measured once weekly at weeks 0 (before), 1, 2, 3 (during) and 4 (after L-glutamine administration). The haematological, RT, HR and RR data obtained weekly were analysed using repeated-measures one-way ANOVA, followed by Tukey's post-hoc test to evaluate differences between periods, and between treated and control groups. The PCV, haemoglobin concentration and RBC count were higher (P < 0.05) in the treated group compared to the control group during the period of L-glutamine administration. These differences were sustained till week 4. Beginning from week 1 of the study, the total leucocyte count in treated group (10.10 ± 0.25 × 10³/μL) was higher (P < 0.05) than the count in control group (7.23 ± 0.41 × 10³/μL), this trend was also maintained throughout the study. The neutrophil:lymphocyte ratio during weeks 3 and 4 of the experiment was lower (P < 0.05) in the treated compared to the control group. RT was lower (P < 0.05) in treated group than the control group. In conclusion, L-glutamine administration ameliorated the adverse effects of heat stress on the haematological parameters in RSGs during the hot-dry season.     

Effect of enzyme extracts isolated from white-rot fungi on chemical composition and in vitro digestibility of wheat straw

A series of in vitro experiments were completed to evaluate the potential of enzyme extracts, obtained from the white-rot fungi Trametes versicolor (TV1, TV2), Bjerkandera adusta (BA) and Fomes fomentarius (FF), to increase degradation of cell wall components of wheat straw. The studies were conducted as a completely randomized design and analysed using one-way ANOVA. Enzyme activities of the extracts, previously obtained from a liquid culture medium, were characterized in terms of laccase and peroxidase for ligninolytic activity. Carboxymethyl cellulase (CMCase) and avicell digesting cellulase (Avicelase) were used for cellulolytic enzyme assays. Wheat straw samples were incubated with enzyme extracts in a citrate buffer (pH 5.0) in a forced air oven at 25 °C for 6 days. In vitro NDF digestibility (IVNDFD), and the rate and extent of NDF fermentation, without and after incubation with the white-rot enzyme extracts, were determined using a gravimetric microbiological method and a gas production technique, respectively. Results from cell wall chemical composition showed that TV2 and BA enzyme extracts decreased NDF concentration (P<0.05) and that TV1 had higher activity (P<0.05) towards cellulose. There was an increase in IVNDFD (P<0.05), resulting from treatment of wheat straw with enzyme extracts from BA, TV1 and TV2, reaching a difference of 13% for TV2 (P<0.05), versus the non-treated straw control. Treatment with enzyme extract from TV2 caused increased gas production (P<0.05) after the first 20 h of incubation, and also increased the maximum rate of gas production, thus enhancing fermentation kinetics. This study indicates that enzyme extracts from white-rot fungi can be used to develop new approaches to overcome low digestibility of some plant cell walls. Utilization of different substrates to produce enzyme extracts can lead to production of viable ligninolytic complexes which could improve the nutritive value of fibrous feeds.     

Amounts of tissue magnesium and some trace elements in cats with mammary tumors related to various clinicopathological parameters

BackgroundMammary tumors are one of the major malignancies seen in cats. Researchers have indicated the similarity between the epidemiological and clinicopathological patterns of feline mammary tumors and human breast cancer (HBC). In recent years, the investigation of trace elements in cancer tissues becomes prevalent in HBC due to the role of these elements in biochemical and physiological processes. This study, it is aimed to evaluate some trace elements in feline mammary tumors according to clinical and pathological findings.MethodsA total of 60 tumoral masses from 16 female cats with mammary tumors were included in the study. The study groups were formed according to histopathology as malignant epithelial tumor (MET; n = 39) and hyperplasia and dysplasia (H&D; n = 21). Copper (Cu), Iron (Fe), Magnesium (Mg), Manganese (Mn), Selenium (Se) and Zinc (Zn) trace elements in mammary tissues were analyzed by using an inductively coupled plasma-optical emission spectrophotometer.ResultsThe mean age and weight of the cats were 11.75 ± 0.75 years and 3.35 ± 0.21 kg; respectively. Eleven of 16 cats were intact whereas the rest of them had been spayed. Metastases were observed in 10 cats. Tissue Mg level in group MET was significantly higher than in group H&D (P < 0.01) while the other elements had not significant differences between the groups. In group MET, analyzed elements were not statistically significant related to the inflammation, ulceration and invasion to the peripheral muscle (P > 0.05). However, tissue Fe level was significantly higher in T2 than in T3 (P < 0.05). The mean levels of tissue Fe, Mg and Mn had significant differences related to histological grading as P < 0.01, P < 0.05 and P < 0.001; respectively. A mild to severe correlation was found between tissue Zn and Se, Cu, Fe, Mg, and Mn levels.ConclusionTissue Mg and some trace elements were evaluated in feline mammary tumours in regard to various clinicopathological parameters. Tissue Mg level was sufficient to differentiate the malignant epithelial tumors from hyperplasia and dysplasia. However, Mn and Se tended to distinguish different tumor types. Tissue Fe, Mg and Mn had significant differences related to histological grading. Also, the Fe level was significantly higher in T2 than in T3 and Zn level tended to be higher in T3 than in T1. It was concluded that Mg, Se, Mn, Fe, Cu and Zn provided useful information on the pathogenesis of feline mammary tumors. Further research is needed on the tissue and serum concentrations of trace elements which may provide valuable information for the disease prognosis.     

Regulation of transporter expression in mouse liver, kidney, and intestine during extrahepatic cholestasis

It is hypothesized that during cholestasis, the liver, kidney, and intestine alter gene expression to prevent BA accumulation; enhance urinary excretion of BA; and decrease BA absorption, respectively. To test this hypothesis, mice were subjected to either sham or bile-duct ligation (BDL) surgery and liver, kidney, duodenum, ileum, and serum samples were collected at 1, 3, 7, and 14 days after surgery. Serum total BA concentrations were 1-5 μmol/l in sham-operated mice and were elevated at 1, 3, 7, and 14 days after BDL, respectively. BDL decreased liver Ntcp, Oatp1a1, 1a5, and 1b2 mRNA expression and increased Bsep, Oatp1a4, and Mrp1-5 mRNA levels. In kidney, BDL decreased Oatp1a1 and increased Mrp1-5 mRNA levels. In intestine, BDL increased Mrp3 and Ibat mRNA levels in ileum. BDL increased Mrp1, 3, 4, and 5 protein expression in mouse liver. These data indicate that the compensatory regulation of transporters in liver, kidney, and intestine is unable to fully compensate for the loss of hepatic BA excretion because serum BA concentration remained elevated after 14 days of BDL. Additionally, hepatic and renal Oatp and Mrp genes are regulated similarly during extrahepatic cholestasis, and may suggest that transporter expression is regulated not to remove bile constituents from the body, but instead to remove bile constituents from tissues.     

Effect of exposure to high temperatures in the excretion of cadmium and lead

ObjectiveThis study aims to observe the effect on urine and sweat excretion levels of cadmium (Cd) and lead (Pb) in healthy men in a maximum incremental test until exhaustion and repeated exposure to heat.Methodstwenty-nine adult men divided into control group (CG; n = 14) and experimental group (EG; n = 15) performing two maximum tests until exhaustion in normothermia (22 °C) and hyperthermia (42 °C). EG experienced 9 sessions of heat exposure at high temperatures (100 °C) (HEHT). After the nine sessions, the initial tests were repeated in both groups. Urine samples were collected before and after each test. After the hyperthermia tests, sweat samples were gathered.ResultsUrinary Cd increased after initial tests in GC and in hyperthermia in EG (p < 0.05). Urinary excretion of Pb rose after HEHT (p < 0.05). Pb in sweat was higher in EG than in CG after HEHT (p < 0.05).ConclusionHeat exercise and constant exposure to heat can be a valid method to increase the excretion of toxic metals.     

Bioactivity of ovulation inducing factor (or nerve growth factor) in bovine seminal plasma and its effects on ovarian function in cattle

To understand the role of ovulation-inducing factor (or nerve growth factor) (OIF [NGF]) in bovine seminal plasma, we (1) used an in vivo llama bioassay to test the hypothesis that bovine seminal plasma induces ovulation and CL development in llamas similar to that of llama seminal plasma when the dose of seminal plasma is adjusted to ovulation-inducing factor content (experiment 1) and (2) determined the effect of bovine seminal plasma on the interval to ovulation and luteal development in heifers (experiment 2). Within species, seminal plasma was pooled (n = 160 bulls, n = 4 llamas), and the volume of seminal plasma used for treatment was adjusted to a total dose of 250 μg of ovulation-inducing factor. In experiment 1, mature female llamas were assigned randomly to four groups and treated intramuscularly with either 10 mL of PBS (negative control, n = 5), 50-μg GnRH (positive control, n = 5), 6-mL of llama seminal plasma (n = 6), or 12 mL of bull seminal plasma (n = 6). Ovulation and CL development were monitored by transrectal ultrasonography. In experiment 2, beef heifers were given a luteolytic dose of prostaglandin followed by 25-mg porcine LH (pLH) 12 hours later to induce ovulation. Heifers were assigned randomly to three groups and given 12 mL bovine seminal plasma intramuscularly 12 hours after pLH treatment (n = 10), within 4 hours after ovulation (n = 9), or no treatment (control, n = 10). Ovulation was monitored by ultrasonography every 4 hours, and the CL development was monitored daily until the next ovulation. In experiment 1, ovulation was detected in 0/5, 4/5, 4/6, 4/6 llamas in the PBS, GnRH, llama seminal plasma, and bovine seminal plasma groups, respectively (P < 0.05). Luteal development was not different among groups. In experiment 2, the interval to ovulation was more synchronous (range: 4 vs. 22 hours; P < 0.0001) in heifers treated with seminal plasma before ovulation compared with the other groups. Luteal development was not different among groups; however, plasma progesterone concentrations tended to be greater in the postovulation treatment group compared with other groups. In summary, results confirmed the presence of bioactive ovulation-inducing factor in bull seminal plasma and supported the hypothesis that bovine and llama seminal plasma have similar ovulatory effects, using a llama bioassay. Treatment with bovine seminal plasma resulted in greater synchrony of ovulation in heifers pretreated with pLH. Plasma progesterone concentration tended to be higher in heifers given bovine seminal plasma within 4 hours after ovulation, suggesting that bovine ovulation-inducing factor is luteotrophic.     

Importance of intense male sexual behavior for inducing the preovulatory LH surge and ovulation in seasonally anovulatory female goats

The present study was carried out to determine whether the presence of photostimulated sedated male goats could stimulate the LH preovulatory surge and ovulation in seasonal anestrous goats. Sexually experienced male goats were treated with artificial long days (16 hours light per day) from 1 November to 15 January to stimulate their sexual activity in March and April, corresponding to the natural sexual rest. A female group of goats (n = 20) was exposed to non-sedated males who displayed an intense sexual behavior and provided strong odor (non-sedated group). Another female group of goats (n = 20) was exposed to the photo-stimulated male goats, but these males were sedated with Xylazine 2% to prevent the expression of sexual behavior (sedated group). The sedated males also provided a strong odor. Females of both groups had full physical and visual contact with non-sedated or sedated males. In both groups, the males remained with females during 4 days. The LH preovulatory surge of 10 female goats per group was measured by determination of LH plasma concentrations in samples taken every 3 hours. In addition, in all goats, (n = 20 by group), ovulation was determined by measuring plasma concentrations of progesterone. The proportion of female goats showing a preovulatory LH surge was higher in goats exposed to non-sedated (10/10) than in those exposed to sedated bucks (0/10; P < 0.0001). Similarly, most of does in contact with non-sedated males ovulated (19/20), but none of those in contact with sedated males did so (0/20; P < 0.0001). We conclude that the expression of an intense sexual behavior by male goats is necessary to induce LH preovulatory surge and ovulation in seasonally anovulatory goats.     

Enhancement of tendon-bone healing with the use of bone morphogenetic protein-2 inserted into the suture anchor hole in a rabbit patellar tendon model

Background aimsSuture anchor fixation failure has been reported as a result of anchor loosening and migration during the tendon-bone repair. The aim of this study was to evaluate the effects of bone morphogenetic protein-2 (BMP-2) inserted into the suture anchor hole on bone formation and the tendon-bone healing.MethodsBoth back legs of 24 New Zealand White rabbits (n = 48) were used in this study. A metal suture anchor was then placed 5 mm below the cortex. In the control group, the space over the eyelet of the anchor (suture anchor hole) was not filled. In the experimental group, the suture anchor hole was filled with 0.1 mL of fibrin glue (group 2) or collagen gel (group 3) with 1 μg BMP-2. Histologic analysis, real-time-polymerase chain reaction, bone density and failure load measurement were performed, and differences were analyzed at 4 and 8 weeks.ResultsHistologic analysis revealed more abundant new bone, mature bone and organized fibrocartilage at the tendon-bone interface at 4 and 8 weeks in groups in which BMP-2 was applied. At 8 weeks, the failure load of groups 1, 2 and 3 was significantly different among the three groups (P = 0.01). After post hoc Tukey test, the failure load of group 2 was significantly higher than that of group 1 (P = 0.01).ConclusionsBMP-2, administrated as described in this study, improved tendon-bone healing and bone formation, resulting in improved biomechanical strength of the tendon-bone junction.     

Determination of experimental domain factors of polyphenols,phenolic acids and flavonoids of lemon (Citrus limon) peel using two-level factorial design

This study aimed to evaluate the significant extraction factors in achieving higher recovery yield of total polyphenols, phenolic acids and flavonoids content from Citrus limon peel using two-level factorial design. The effect of five independent factors including drying temperature (40–60 °C), methanol concentration (20–60%), extraction temperature (28–60 °C), extraction time (30–60 min) and storage duration (0–14 days) were evaluated. Among all the examined factors, results showed that drying temperature, storage duration and extraction temperature were the most significant and contributing factors affecting the total polyphenols, phenolic acids and flavonoids content of lemon peel at P < 0.05. On the contrary, methanol concentration and extraction time exhibited the least significant and contribution at P greater than 0.05. In conclusion, the experimental domain factors were successfully obtained from this experiment, Therefore, further study on optimization of the obtained factors will be conducted in the future study using response surface methodology.