Comparison of in vitro and in vivo fertilizing potential of buffalo bull semen frozen in egg yolk-, soya bean lecithin- and liposome-based extenders.

The objective of this study was to compare different extenders for post-thaw in vitro sperm function and in vivo fertility of buffalo semen. Accordingly, sperm of 30 ejaculates extended in egg yolk (TRIS with 20% egg yolk; EY), two soya lecithin-based (SL-1; AndroMed® and SL-2; Bioxcell® ) and a liposome-based extender (LS; OptiXcell® ) were tested. The post-thaw semen was evaluated for computer-assisted sperm analysis (CASA), sperm viability, membrane and acrosome integrity, DNA integrity and acrosome reaction and first service pregnancy rate (FSPR) in a fixed-time artificial insemination programme. Total motility and VCL were the only CASA-based parameters that exhibited significantly higher (p < .05) percentage in LS among these extenders. Post-thaw percentage of acrosome integrity (55.9 ± 1.4, 58.1 ± 2.0, 55.8 ± 2.0, 56.6 ± 2.3) and DNA integrity (68.8 ± 2.0, 69.2 ± 2.3, 71.3 ± 2.1, 69.1 ± 2.1) did not differ (p > .05) in EY, SL-1, SL-2 and LS extender, respectively. However, a variable response in terms of efficacy of different extenders for sperm viability and plasma membrane integrity was observed. Assessment of inducibility of acrosome reaction showed significant differences between extenders (51.9 ± 2.1, 44.3 ± 2.4, 46.1 ± 2.3 and 58.1 ± 3.1%, respectively, for EY, SL-1, SL-2 and LS). Furthermore, field trials revealed significantly higher (p < .05) FSPR of LS-extended semen as compared to that for EY, SL-1 and SL-2 extender (46.3%, 41.2%, 31.2% and 29.7%, respectively). It is concluded that the liposome-based extender is more effective than egg yolk- and soya lecithin-based extenders and may be used for cryopreservation of buffalo semen in the future.

Birth weight, early life weight gain and age at menarche: a systematic review of longitudinal studies.

BACKGROUND AND OBJECTIVE: Adiposity in pre- and postnatal life may influence menarcheal age. Existing evidence is primarily cross-sectional, failing to address temporality, for which the role of adiposity in early life remains unclear. The current study sought to systematically review longitudinal studies evaluating the associations between birth weight and infant/childhood weight status/weight gain in relation to menarcheal age. METHODS: PubMed, EMBASE, Web of Science, Global Health (Ovid) and CINAHL were systematically searched. Selected studies were limited to English-language articles presenting multi-variable analyses. Seventeen studies reporting risk estimates for birth weight (n = 3), infant/childhood weight gain/weight status (n = 4) or both (n = 10), in relation to menarcheal age were included. RESULTS: Lower vs. higher birth weight was associated with earlier menarche in nine studies and later menarche in one study, while three studies reported a null association. Greater BMI or weight gain over time and greater childhood weight were significantly associated with earlier menarche in nine of nine and six of seven studies, respectively. CONCLUSIONS: Studies suggested that lower birth weight and higher body weight and weight gain in infancy and childhood may increase the risk of early menarche. The pre- and postnatal period may thus be an opportune time for weight control interventions to prevent early menarche, and its subsequent consequences.

Review on mechanisms of dairy summer infertility and implications for hormonal intervention.

In dairy cows and buffaloes, summer heat stress (HS) reduces milk yield and delays return to pregnancy leading to financial loss. Clues for effective interventions against summer infertility (SI) lie in understanding the underlying mechanisms. This article reviews current knowledge on the mechanisms of bovine SI and their implication for hormonal management. Under HS dairy animals encounter anestrous, silent cycles and repeat breeding which extend their open period. These effects are attributed mainly to HS induced disturbances in luteinizing hormone (LH) secretion, follicular dominance and estrogen secretion, ovulation and oocyte competence, luteal development and progesterone secretion, utero-placental function and embryo-fetal development. Hormonal timed artificial insemination protocols and LH support around estrous improved summer pregnancy rates by avoiding need for estrus detection, assisting follicular development and ovulation, enhancing quality oocytes and stimulating luteal function. Progesterone supplementation to enhance embryonic development did not produce significant improvement in summer pregnancy rates. There is need for evaluating integrated approaches combining hormones, metabolic modifier and cyto-protective agents.

Comparison of follicular dynamics, superovulatory response, and embryo recovery between estradiol based and conventional superstimulation protocol in buffaloes (Bubalus bubalis).

AIM: To evaluate the follicular dynamics, superovulatory response, and embryo recovery following superstimulatory treatment initiated at estradiol-17ß induced follicular wave emergence and its comparison with conventional superstimulatory protocol in buffaloes. MATERIALS AND METHODS: Six normal cycling pluriparous buffaloes, lactating, 90-180 days post-partum, and weighing between 500 and 660 kg were superstimulated twice with a withdrawal period of 35 days in between two treatments. In superstimulation protocol-1 (estradiol group) buffaloes were administered estradiol-17ß (2 mg, i.m.) and eazibreed controlled internal drug release (CIDR) was inserted intravaginally (day=0) at the random stage of the estrous cycle. On the day 4, buffaloes were superstimulated using follicle stimulating hormone (FSH) 400 mg, divided into 10 tapering doses given at 12 hourly intervals. Prostaglandin F2α analogs (PGF2α) was administered at day 7.5 and day 8, and CIDR was removed with the second PGF2α injection. In superstimulation protocol - 2 (conventional group) buffaloes were superstimulated on the 10(th) day of the estrous cycle with same FSH dose regimen and similar timings for PGF2α injections. In both groups, half of the buffaloes were treated with luteinizing hormone (LH) 25 mg and other half with 100 ug buserelin; gonadotrophin releasing hormone (GnRH) analog at 12 h after the end of FSH treatment. All buffaloes in both protocols were inseminated twice at 12 and 24 h of LH/GnRH treatment. Daily ultrasonography was performed to record the size and number of follicles and superovulatory response. RESULTS: Significantly higher number of small follicles (<8 mm) was present at the time of initiation of superstimulatory treatment in the estradiol group compared to the conventional group (12.5±0.80 vs. 7.3±1.21, respectively, p=0.019), however, the number of ovulatory size follicles (≥8 mm) did not differ significantly between the respective groups (15.5±1.24 vs. 12.2±1.30; p=0.054). Total embryos and transferable embryos recovered were non-significantly higher in the estradiol group compared to the conventional group (5.83±0.86 vs. 4.67±1.16, p=0.328, and 3.67±0.93 vs. 2.67±0.68, p=0.437, respectively). The significant higher proportion of transferable embryos were recovered in buffaloes treated with LH compared to GnRH (73.3% vs. 48.5%; p=0.044). CONCLUSION: The average number of ovulatory size follicles (>8 mm), corpora lutea, and transferable embryos was higher in buffaloes superstimulated at estradiol-induced follicular wave compared to the conventional protocol: Further the percentage of transferable embryos was significantly higher in buffaloes administered with LH compared to GnRH.

Correlation of blood flow of the preovulatory follicle to its diameter and endocrine profile in dairy buffalo.

Blood flow of the preovulatory follicle (POF) wall can be used as a predictor of the quality of POF. Our aim was to determine the correlation of blood flow of POF with the POF diameter, and intra-follicular and plasma concentrations of Insulin-like Growth Factor-I (IGF-1) and oestradiol in dairy buffalo. Nine Murrah buffalo subjected to an ovulation synchronization protocol (Ovsynch) were assessed on day 10 of the protocol for diameter and blood flow of POF, followed by the aspiration of follicle fluid. Prior to follicular aspiration, blood samples were obtained from jugular vein for estimation of IGF-1 and oestradiol. The vascularity of POF was determined (Range: 250-967 pixel(2) ) along with intra-follicular and plasma concentration of IGF-1 (Range: 9.3-31.8 ng/ml and 14.7-29.7 ng/ml respectively) and oestradiol (Range: 124.2-447.9 ng/ml and 0.25-1.05 ng/ml respectively). Diameter of the POF was weakly correlated (r = 0.21, p < 0.01) with blood flow to it. As compared to POF diameter, the blood flow of POF had greater positive correlation with intra-follicular and plasma concentrations of hormones (IGF-1 and oestradiol). A strong positive correlation was recorded between intra-follicular IGF-1 and oestradiol. Also, plasma concentrations of oestradiol and progesterone were negatively correlated In brief, assessment of the blood flow of the POF is a non-invasive and reliable indicator of its functional competence as compared to the POF diameter.

Uterine blood flow during various stages of pregnancy in dairy buffaloes using transrectal Doppler ultrasonography.

Reference data to assess function and perfusion of uteroplacental tissues by assessment of uterine blood flow parameters is not available in buffaloes. The objective of the present study was to assess and quantify blood flow parameters at various stages of gestation. A total of 54 Murrah buffaloes, six animals at every month of gestation, from two months to ten months, were subjected to transrectal Doppler ultrasonography to assess uterine blood flow in both the middle uterine arteries. Resistance and pulsatility indices, velocity and volume of blood flow, diameter of the artery and notch signal were measured for different waveforms. The resistance index value was positively correlated with pulsatility index (r=0.94, P<0.05) but negatively correlated with all other parameters for both arteries, ipsilateral and contralateral to the fetus. With a significant advancement of pregnancy, there was reduction in resistance index with a significant increase in velocity, volume of blood flow and diameter in both arteries. There was a significant increase in the volume and velocity of blood flow (P<0.05) during the last trimester with increased growth demands of the fetus. The notch signal disappeared by 20-24 weeks. Thus, transrectal Doppler ultrasongraphy proved to be a useful non-invasive method to assess the uterine blood flow during pregnancy in buffaloes. This could be a valuable tool to ascertain hemodynamic changes in complicated pregnancy (e.g. abnormalities in uterine blood flow/placenta/fetus) and its extent of deviation from normal.

Attenuation of luteolytic response following fish meal supplementation in dairy buffaloes (Bubalus bubalis).

Luteolysis of corpus luteum, due to un-inhibited PGF(2α) secretion, has been reported to be a cause of early embryonic mortality in dairy animals. The objective of this study was to determine the effects of fish meal (FM) supplementation on the uterine secretion of PGF(2α) and hence establish its supplementation as an antiluteolytic strategy in dairy buffaloes. Five cycling Murrah buffaloes were supplemented with 250g FM daily for 55 days in addition to their routine feed and seven buffaloes were kept as non-supplemented control. After 30 days of FM supplementation, the oestrus was synchronized in all the buffaloes using Ovsynch protocol. On day 15 of synchronized cycle, animals were challenged with oxytocin (OT; 100IU) intravenously and blood samples were collected at 15min interval, 1h before to 4h after OT challenge. The PGF(2α) response was measured as the venous concentration of 13,14-dihydro-15-keto PGF(2α) (PGFM). The mean hourly concentration of PGFM in FM supplemented buffaloes was lower than in the control buffaloes at all the occasions. During peak response (1h post-OT challenge), PGFM concentration was significantly lower (P<0.05) in FM supplemented buffaloes than in the control (197.4±41.7pg/ml versus 326.3±33.5pg/ml, respectively). Also the percent rise in PGFM after OT-challenge in FM supplemented buffaloes was less than the control (11.73% versus 22.47%). The dietary supplementation did not affect the size of corpus luteum (CL) and plasma progesterone concentration. Plasma glucose and total protein concentrations remained within the normal physiological limits during FM supplementation. The present study indicated that supplementing FM decreased the concentrations of PGF(2α) without alterations in the size of CL and plasma progesterone concentrations in dairy buffaloes.

Postpartum ovarian activity in South Asian zebu cattle.

Timely onset of postpartum ovarian activity is vital for optimal reproductive performance of dairy cows. Much depends upon genetic constitution of an animal although several factors interplay to govern the onset of postpartum ovarian activity. South Asian zebu cattle have much longer service period when compared with other exotic or crossbred cattle reared in the same Asian environment, which suggests differences in their genetic makeup. However, the cows with same genetic configuration expressed better reproductive potential when reared under different environment, such as in Brazil and Mexico, which suggests the role of extrinsic factors such as management, nutrition, environment and disease conditions. Better management of animals (provision of proper shade, water and housing, efficient oestrous detection and timely insemination), good quality nutrition supplemented with appropriate minerals and vitamins, prevention of diseases (vaccination, deworming, suitable therapeutic interventions) and application of biotechnology have helped in improving postpartum ovarian activity and, therefore, reproductive performance of zebu cattle in Asia. No comprehensive study appears to have been carried out on the various aspects of reproduction in zebu cattle reared under South Asian socio-agro-climatic conditions. This paper is a modest effort to collect what ever information available and to critically review the postpartum ovarian activity in zebu cattle with special reference to the effect of the various managemental practices and pharmacological interventions.

Lack of correlation of degree of villous atrophy with severity of clinical presentation of coeliac disease.

BACKGROUND AND AIM: Both the clinical presentation and the degree of mucosal damage in coeliac disease vary greatly. In view of conflicting information as to whether the mode of presentation correlates with the degree of villous atrophy, we reviewed a large cohort of patients with coeliac disease. PATIENTS AND METHODS: We correlated mode of presentation (classical, diarrhoea predominant or atypical/silent) with histology of duodenal biopsies and examined their trends over time. RESULTS: The cohort consisted of 499 adults, mean age 44.1 years, 68% females. The majority had silent coeliac disease (56%) and total villous atrophy (65%). There was no correlation of mode of presentation with the degree of villous atrophy (p=0.25). Sixty-eight percent of females and 58% of males had a severe villous atrophy (p=0.052). There was a significant trend over time for a greater proportion of patients presenting as atypical/silent coeliac disease and having partial villous atrophy, though the majority still had total villous atrophy. CONCLUSIONS: Among our patients the degree of villous atrophy in duodenal biopsies did not correlate with the mode of presentation, indicating that factors other than the degree of villous atrophy must account for diarrhoea in coeliac disease.

Enhancing reproductive performance in dairy buffalo: major constraints and achievements.

Buffalo are of high economic importance for farmers in several developing countries but reproductive performance is poor. A large proportion of heifers attain puberty at 3-5 years of age. A good quality diet supplemented with extra nutrients reduces the age of puberty, whereas the effects of administration of exogenous GnRH or equine chorionic gonadotrophin (eCG) are equivocal. The incidence of anoestrus in buffalo ranges from 20 to 80% depending on season. Most buffalo cease ovarian cyclicity during hot summers probably due to the combined effects of nutrition, environment and management. Keeping buffalo cool by wallowing, water sprinklers or shade improves fertility. Supplementary feeding with Urea Molasses Multi-nutrient Blocks (UMMB) for 60 days before calving enhances the early onset of postpartum oestrus. Regular UMMB supplementation also improves pregnancy rates in anoestrous non-pregnant buffalo. Prepartum vaginal prolapse is hereditary and eradication can be achieved by genetic selective breeding programmes. Treatment with calcium, phosphorus and progesterone gives only transient relief to clinical cases. Uterine torsion is the most common cause of dystocia (70%). Deployment of Sharma's detorsion method and anti-stress measures increase survival rates in cases presented within 36 h. In conclusion, greater understanding about the effects of better year-round nutrition, improved management and markers for logical breeding programmes are essential to curtail the incidence of the reproductive disorders that reduce buffalo fertility.

Aggression and schizophrenia: efficacy of risperidone.

The advent of novel antipsychotic medications has raised treatment expectations for patients with severe mental illness. In this regard, clozapine has been particularly effective in reducing aggressive behavior in patients with schizophrenia. This study compared the efficacy of risperidone and conventional antipsychotic medications in the management of hostile patients. Improvements in the level of aggression were evident over time in both treatment groups, and a similar response between risperidone and typical antipsychotics was observed. Future studies should address the relative role of typical antipsychotics, adjunctive agents, and novel antipsychotic medications in the pharmacological management of persistent aggression in patients with schizophrenia.

Improved therapeutic window for treatment of histotoxic hypoxia with a free radical spin trap.

The therapeutic time window for N-methyl-D-aspartate (NMDA) antagonists, non-NMDA antagonists, and glutamate release inhibitors in focal models of ischemia appears to be about 1-2 h. In contrast, a free radical spin trap was found to have an improved therapeutic window. We compared the therapeutic time windows of the NMDA antagonist dizolcilpine maleate (MK-801), the glutamate release inhibitor lamotrigine, and the free radical spin trap n-tert-butyl-alpha-(2-sulfophenyl)-nitrone (S-PBN) against striatal lesions produced by the mitochondrial toxin malonate, which produces histotoxic hypoxia. Lamotrigine exerted neuroprotective effects when administered at 1 h before malonate injections. MK-801 protected at 1 h before and 1 h after malonate injections, whereas S-PBN showed efficacy when administered up to 6 h after malonate injections. Striatal injections of malonate produced a rapid increase in lactate production and early changes in diffusion-weighted imaging as assessed by magnetic resonance imaging. Therefore, the time course to evolve a lesion in our model of histotoxic hypoxia is comparable with that of other models of focal ischemia. These findings provide direct evidence that a free radical spin trap has an improved therapeutic window compared to an NMDA antagonist and a glutamate release inhibitor. This could be a therapeutic advantage in the treatment of clinical stroke patients.

Further characterization of the N-terminal copper(II)- and nickel(II)-binding motif of proteins. Studies of metal binding to chicken serum albumin and the native sequence peptide.

We have investigated the Cu(II)- and Ni(II)-binding properties of chicken serum albumin (CSA) and of the native sequence tripeptide derived from the N-terminus of this protein. Spectrophotometric and equilibrium dialysis experiments demonstrate that Cu(II) and Ni(II) bind non-specifically at the N-terminus of CSA. Proton displacement studies show that the histidine residue in the fourth position of the protein does not appear to participate in the binding of the two metals. Consistent results were obtained with the native sequence tripeptide L-aspartyl-L-alanyl-L-glutamic acid N-methylamide. The results presented here demonstrate that neither the glutamic acid residue in the third position nor the histidine in the fourth position participate in the binding of Cu(II) and Ni(II) to CSA. It is known, however, that a number of other albumins with a histidine residue in the third position possess high-affinity Cu(II)- and Ni(II)-binding sites. Our results provide further evidence that the N-terminal Cu(II)/Ni(II)-binding motif requires a histidine at the third position in order to bind Cu(II) and Ni(II) specifically.

Inhibition of cell-cell adhesion and morphogenesis of Dictyostelium by carnitine.

Carnitine (gamma-trimethylammonium beta-hydroxy-butyric acid) possesses the novel property of preventing cell aggregation elicited by clusterin or by fibrinogen (I.B. Fritz and K. Burdzy, J. Cell. Physiol., 140:18-28 [1989]). In investigations reported here, we show that carnitine also affects cell-cell adhesion in Dictyostelium discoideum, a cellular slime mold whose cells interact in specific and complex manners during discrete stages of development. Two types of cell adhesion systems sequentially appear on the surface of developing Dictyostelium cells, involving the surface glycoprotein gp24 which mediates EDTA-sensitive binding sites, and the surface glycoprotein gp80 which mediates the EDTA-resistant binding sites. Addition of increasing concentrations of D(+)-carnitine and L(-)-carnitine resulted in a progressive inhibition of both the EDTA-sensitive binding sites and the EDTA-resistant binding sites of Dictyostelium cells at different stages of development. In contrast, comparable or higher concentrations of choline, acetyl-beta-methylcholine, or deoxycarnitine had no detectable effects on cell aggregation. Concentrations of carnitine required for 50% inhibition of EDTA-resistant adhesion sites were found to be dependent upon levels of gp80 expressed by Dictyostelium, with greatest inhibition by carnitine of reassociation of cells containing the lowest levels of gp80. Removal of carnitine from cells by washing resulted in the rapid restoration of the ability of Dictyostelium to form aggregates and to resume normal development. We discuss possible mechanisms by which carnitine inhibits the aggregation of cells.

Repeated dosing of Uniphyl tablets under fed and fasting conditions: comparison of serum theophylline levels, pulmonary function, and asthma symptoms.

In order to determine the effects of repeated administration under fed and fasting conditions on the bioavailability and clinical efficacy of Uniphyl tablets, 22 adult asthmatics took the drug immediately following their evening meal for seven consecutive days and under fasting conditions for an additional seven consecutive days. For each patient, the daily theophylline dose remained constant throughout the study. Peak and trough serum theophylline concentrations (STC), spirometry, asthma symptoms, side effects and use of beta-agonist inhalers were recorded daily at 0730 and 1900 hours. The mean daily theophylline dose was 818.2 +/- 213.0 mg. The mean peak STC when Uniphyl was taken with food was 14.4 +/- 4.5 mg/L and was 13.1 +/- 3.6 mg/L when taken under fasting conditions (P less than .05). The trough STC was 7.4 +/- 2.8 mg/L with food and 6.9 +/- 2.1 mg/L while fasting (NS). There were no significant differences between the two dosing conditions in terms of spirometry, asthma symptom scores, side effects or use of beta-agonist inhalers. There was no significant difference between the patients' morning and evening FEV1 under either dosing condition. Since the differences in STC between fed and fasting conditions were not clinically significant, we conclude that there is no need to restrict patients to a rigid relationship between Uniphyl dosing and meal conditions. On the basis of patient preference and compatibility with a normal lifestyle, we recommend that patients should generally be instructed to take the drug with or shortly following their evening meal.