Antisperm antibodies in repeat-breeding cows: Frequency, detection and validation of threshold levels employing sperm immobilization, sperm agglutination and immunoperoxidase assay.

Antisperm antibodies have been found in repeat-breeding(RB) cows, and those causing agglutination and/or immobilization of sperm are considered to be closely related to unexplained infertility. However, a standard protocol for identifying antisperm antibodies (ASA) in cattle is not validated. Therefore, an investigation was undertaken to evaluate sperm immobilization (SIT), sperm agglutination (SAT) and immunoperoxidase (IPT)assays for detection of ASA in serum and their respective threshold levels for confirmation. Animals (heifers, normally breeding, repeat-breeding and pregnant animals) that were free from IBR, brucellosis and uterine infections (screened by clinical examination) were included in the study. Sperm agglutinating, sperm immobilizing and antisperm antibodies evaluated by respective assay were significantly higher (p < .05) in RB cows compared to other groups. The SIT assay was able to identify 61% of RB caused by ASA, more than those employing SAT and IPT. Furthermore, a dilution rate of 1:5 and 1:80 (confirms 59.0 and 57.0% RB+ve)were sufficient to diagnose ASA by SAT and IPT, respectively. Results indicate the presence of __12.6% clumped spermatozoa and __ 2.6%(cut-off value) peroxidase-positive spermatozoa at 1:5 and 1:80 dilutions diagnosed with SAT and IPT, respectively, may be considered as repeaters arising out of ASA. Furthermore, study also showed the presence of lower incidence of ASA positivity in other groups of animals (heifer

Supplementation of Slow-Release Melatonin Improves Recovery of Ovarian Cyclicity and Conception in Summer Anoestrous Buffaloes (Bubalus bubalis).

The role of melatonin as a protective neurohormone against restoring cyclicity in summer anoestrous animals in photoperiod species has gained wider acceptance. This study was designed to uncover the evidence the slow-release melatonin (MLT) has on initiation of ovarian cyclicity and conception rate (CR) in summer anoestrous buffaloes. Thus, buffaloes diagnosed as summer anoestrous (absence of overt signs of oestrus, concurrent rectal examination and radioimmunoassay for serum progesterone at 10 days interval) were grouped as untreated (Group I, sterilized corn oil, n = 8) and treated (Group II, single subcutaneous injection of MLT @18 mg/50 kg bwt in sterilized corn oil, n = 20). Animals treated and detected in oestrus were artificially inseminated (AI) followed by division into Group III (second dose of MLT on 5th day post-AI, n = 8) and Group IV (no melatonin administration, n = 10). Blood samples were collected at 4 days interval for estimation of serum MLT, progesterone and oestrogen using radioimmunoassay kit. Mean oestrous induction rate (OIR), oestrous induction interval (OII), interoestrous interval (IOI) and CR were estimated. Compared to control, concentration of melatonin was significantly (p < 0.05) higher in treated group ranging from 14.34 ± 1.72 to 412.31 ± 14.47 pg/ml whereas other two hormones did not show any concentration difference. Melatonin-administered buffaloes showed significantly (p < 0.05) higher (90%) OIR with OII of 18.06 ± 1.57 days. Results showed improvement in conception rate in buffaloes administered with post-insemination melatonin. It can be concluded from the study that slow-release melatonin supplementation restored cyclicity in summer anoestrous animals resulting in improvement in conception rate in buffaloes.

A prospective double blind study comparing the effects of oxytocin and human chorionic gonadotrophin as trigger for ovulation.

This study was undertaken to compare the effect of oxytocin and human chorionic gonadotrophin (hCG) on ovulation and conception. A total of 100 women with anovulatory infertility were allocated into two equal groups to take clomiphene citrate plus hCG or clomiphene citrate plus oxytocin. The size and number of follicles by transvaginal sonography determined the administration of hCG or oxytocin. The serum progesterone concentration was measured to provide evidence for ovulation. The ovulation and pregnancy rate of the two groups were compared. There was no significant difference between the groups with respect to mean number of follicles, mean follicular size and rate of ovulation in three cycles. The rate of pregnancy was higher in the oxytocin group compared with the hCG group (12% vs 4%) but it failed to reach statistical difference (p = 0.140). Mild pain was the only side-effect observed in the two groups. Oxytocin was found to be a viable alternative to hCG for triggering ovulation in infertile patients.

Sequestration of PDC-109 protein by specific antibodies and egg yolk cryoprotects bull spermatozoa.

PDC-109, one of the most abundant proteins in bovine seminal plasma, has detrimental effect on spermatozoa in a time- and concentration-dependent manner. Therefore, we hypothesized that sequestration of detrimental protein from ejaculates would be beneficial following cryopreservation of sperm cells. To this aim, we evaluated the effect of sequestration of PDC-109 either by anti-PDC-109 antibodies (Ab) or egg yolk (EY) alone or by the synergistic action of EY + Ab in minimizing cryoinjury to bull spermatozoa. PDC-109 protein was purified by applying two-step chromatography procedures. The purified protein was injected in rabbits to raise antibodies which were isolated using ion-exchange chromatography. After checking the Ab cross-reactivity, they were quantitated and added to ejaculates, either alone or in addition to EY in Tris-glycerol (TG) extender. Thus, ejaculates were processed in extender containing EY + TG (group I), Ab + TG (group II) or EY + Ab + TG (group III). Semen quality parameters (SQPs) viz. viability and acrosome integrity (FITC-PSA), cryoinjury to spermatozoa (chlortetracycline, CTC assay) and in vitro fertility of protein-sequestered-semen (zona-penetration assay) were evaluated. A significant (p < 0.05) improvement in post-thaw SQPs as well as in non-capacitated spermatozoa observed at pre-freeze and post-thaw stages of cryopreservation in group III compared with other groups indicated reduction in protein-mediated cryoinjury. From this study, it can be concluded that sequestration of PDC-109 by synergistic action of EY+Ab as compared to either of them alone significantly improve sperm quality and minimize cryoinjury to bull spermatozoa upon storage at ultra-low temperatures.

Impact of geographic range on genetic and chemical diversity of Indian valerian (Valeriana jatamansi) from northwestern Himalaya.

An effort was made to determine the impact of geographic range on genetic richness and chemical constituents of Valeriana jatamansi Jones, an herb indigenous to the northwestern Himalaya. The genetic structure of 16 accessions from two major divisions of Uttarakhand state (Kumaon and Garhwal) was analyzed by ISSR markers. Overall genetic diversity among the populations was 45 %, with a cumulative range of 35-92 % similarity for most of the high-altitude plants and a comparatively narrow range, 50-88 %, for the population below the altitude of 1,800 m. Likewise, a remarkable predictability was evident from the chemical constituents on an individual basis. In principal component analysis, most of the accessions fall into two major groups and are classified as chemotypes based on the percentage of similar chemical constituents; these are mostly correlated to altitude. Geographic distance seems to influence the genetic and chemical variability, indicating the genetic inbreeding within the population.

Atypical mucocutaneous involvement with Leishmania donovani.

Mucocutaneous leishmaniasis has rarely been reported from India. The usual causative organisms of this infection are Leishmania braziliensis and L. tropica. Another species, L. donovani, which usually causes visceral leishmaniasis, has recently been reported to cause mucocutaneous disease in a few patients from Sri Lanka. We report two patients who had undiagnosed chronic skin lesions for several years. Skin biopsies revealed Leishmania and the species was characterized as L. donovani in both patients. There was considerable improvement in the skin lesions following treatment with liposomal amphotericin B.

A prospective double blind trial investigating impact of vaginal pH on efficacy of prostaglandin gel for cervical ripening and course of labour.

This prospective, double blind investigation was carried out to see the effect of vaginal pH on the efficacy of prostaglandin gel (PGE2) for cervical ripening and course of labour. A total of 45 pregnant women with indications for induction of labour were allocated to two groups: a low vaginal pH (≤5.5, n = 20) and high vaginal pH (>5.5, n = 25) group. All women received prostaglandin E2 gel (0.5 mg in 2.5 ml) intravaginally with repeated dosing if needed, 6 h apart, maximum of three doses. Bishop's score change over 18 h differed significantly between the two groups (p = 0.037). There was no significant difference between the groups with respect to time to onset of labour (9.65±6.29 vs 6.76±3.94, p = 0.066), time to active labour (15.38±9.49 vs 14.30±5.85, p = 0.664), time to complete cervical dilation (18.27±13.85 vs 18.34±6.45; p = 0.984), and time to overall delivery (21.52±9.66 vs 19.39±6.45, p = 0.381).

Polymorphism of glutathione S-transferase M1 and T1 gene loci in COPD.

Chronic obstructive pulmonary disease (COPD) is a complex combination of signs and symptoms in patients with chronic bronchitis and emphysema, diseases that largely result from cigarette smoking. A little information is available for the underlying molecular mechanisms that are responsible for its occurrence. Polymorphisms in genes of xenobiotics metabolizing enzymes are expected to modulate individual responses to genotoxic carcinogens. Present study was a case-control study of COPD patients and healthy controls. Genetic polymorphisms of GSTM1 and GSTT1 genes in 50 COPD patients and 50 healthy controls were investigated using multiplex polymerase chain reaction-restriction fragment length polymorphism techniques to determine whether polymorphisms of these genes are linked to genetic susceptibility to COPD. All subjects were males and smokers. The frequency of GSTM1 homozygous null genotype was 28.0% in COPD cases when compared with controls (32.0%). The difference was not significant showing that risk of COPD was not associated with the GSTM1 null genotypes. The frequencies of homozygous null genotypes of GSTT1 were significantly higher in COPD cases as compared with controls (40% versus 14.0%) suggesting that the theta-glutathione S-transferases null genotype may be associated with the susceptibility to COPD. No significant differences were observed when comparisons were performed according to severity of disease and smoking for GSTM1 and GSTT1. It was also observed that COPD developed in the early age and with a shorter pack-year history in Indian population.

A prospective double blind study using oral versus vaginal misoprostol for labour induction.

This prospective double blind study was undertaken to compare the safety and efficacy of oral vs vaginal misoprostol in equivalent doses (50 microg) for induction of labour. A total of 128 term pregnancies with indication for induction of labour were allocated to two groups to receive 50 microg misoprostol orally or vaginally, every 4 h until adequate contractions were achieved or a maximum of 200 microg dose. Induction to delivery interval was significantly shorter in the vaginal group compared with the oral group (14.6 h vs 22.5 h; p < 0.001). There was no significant difference between the groups with respect to mode of delivery, neonatal outcome and maternal side-effects. However, the incidence of abnormal contractility pattern was more common in the vaginal group (10/68, 14.6%) as compared with the oral group (4/60, 6.6%) (p = 0.146).

In vitro propagation of Rauwolfia serpentina using liquid medium, assessment of genetic fidelity of micropropagated plants, and simultaneous quantitation of reserpine, ajmaline, and ajmalicine.

Rauwolfia serpentina holds an important position in the pharmaceutical world because of its immense anti-hypertensive properties resulting from the presence of reserpine in the oleoresin fraction of the roots. Poor seed viability, low seed germination rate, and enormous genetic variability are the major constraints for the commercial cultivation of R. serpentina through conventional mode. The present optimized protocol offers an impeccable end to end method from the establishment of aseptic cultures to in-vitro plantlet production employing semisolid as well liquid nutrient culture medium and assessment of their genetic fidelity using polymerase chain reaction based rapid amplification of polymorphic DNA analysis. In vitro shoots multiplied on Murashige and Skoog basal liquid nutrients supplemented with benzo[a]pyrene (1.0 mg/L) and NAA (0.1 mg/L) and in-vitro rhizogenesis was observed in modified MS basal nutrient containing NAA (1.0 mg/L) and 2% sucrose. In-vitro raised plants exhibited 90-95% survival under glass house/field condition and 85% similarity in the plants regenerated through this protocol. Field established plants were harvested and extraction of indole alkaloid particularly reserpine, ajmaline and ajmalicine and their simultaneous quantitation was performed using monolithic reverse phase high-performance liquid chromatography (HPLC).

Temporal analysis of meiotic DNA double-strand break formation and repair in Drosophila females.

Using an antibody against the phosphorylated form of His2Av (gamma-His2Av), we have described the time course for the series of events leading from the formation of a double-strand break (DSB) to a crossover in Drosophila female meiotic prophase. MEI-P22 is required for DSB formation and localizes to chromosomes prior to gamma-His2Av foci. Drosophila females, however, are among the group of organisms where synaptonemal complex (SC) formation is not dependent on DSBs. In the absence of two SC proteins, C(3)G and C(2)M, the number of DSBs in oocytes is significantly reduced. This is consistent with the appearance of SC protein staining prior to gamma-His2Av foci. However, SC formation is incomplete or absent in the neighboring nurse cells, and gamma-His2Av foci appear with the same kinetics as in oocytes and do not depend on SC proteins. Thus, competence for DSB formation in nurse cells occurs with a specific timing that is independent of the SC, whereas in the oocytes, some SC proteins may have a regulatory role to counteract the effects of a negative regulator of DSB formation. The SC is not sufficient for DSB formation, however, since DSBs were absent from the heterochromatin even though SC formation occurs in these regions. All gamma-His2Av foci disappear before the end of prophase, presumably as repair is completed and crossovers are formed. However, oocytes in early prophase exhibit a slower response to X-ray-induced DSBs compared to those in the late pachytene stage. Assuming all DSBs appear as gamma-His2Av foci, there is at least a 3:1 ratio of noncrossover to crossover products. From a comparison of the frequency of gamma-His2Av foci and crossovers, it appears that Drosophila females have only a weak mechanism to ensure a crossover in the presence of a low number of DSBs.

Implications of free radicals and antioxidant levels in carcinoma of the breast: a never-ending battle for survival.

BACKGROUND: Under normal circumstances, there is a steady balance between the production of oxygen derived free radicals and their destruction by the cellular antioxidant system inside the human body. However, any imbalance between the levels of these oxidants and antioxidants might cause DNA damage and may lead to cancer development. The aim of this study was to evaluate the level of antioxidants and free radicals in blood and tissue of cancer patients and compare these levels at different TNM stages to derive the possible role of free radicals and antioxidant enzymes in the etiology of breast cancer. MATERIALS AND METHODS: This study includes 30 patients suffering from cancer breast and 20 patients as controls who had benign breast diseases. Circulating lipid peroxide (Malonyldialdehyde [MDA]) levels and activities of the defensive enzymes (Superoxide Dismutase [SOD] and Catalase [CAT]) were estimated in the blood and breast tissue of these patients. RESULTS: Increased levels of free radicals and low levels of antioxidants were observed in malignant tissue. An elevated lipid peroxide concentration was found in the tissue of all the cancer breast patients as evidenced by an increase in the mean MDA level seen with increasing TNM stage of carcinoma breast. Levels of antioxidants SOD and CAT were decreased in cancer patients. CONCLUSION: The results of our study suggest that free radical activity is enhanced in cancer breast patients while the antioxidant defense mechanism is weakened. This activity is enhanced with the increasing severity of cancer as depicted in different TNM stages of breast cancer.

Beyond Beauty : Botulinum Toxin Use in Anal Fissure.

BACKGROUND: Chronic fissure in ano is a common distressing problem with high recurrence rates. Any treatment modality should be simple, effective and reversible with no permanent sequelae. Long term medical management with poor compliance or surgical therapy with risk of incontinence are both less than ideal. In this scenario chemical sphincterotomy using Botulinum toxin offers an alternative modality. METHODS: A total of 30 patients of chronic fissure in ano were treated with Botulinum toxin injection in the internal sphincter. RESULT: All patients had significant symptomatic relief with high rates of fissure healing. The simplicity of administration, lack of complications and cost effectiveness make it a useful alternative to the currently practiced approaches. CONCLUSION: Botulinum toxin offers a simple outpatient treatment of chronic fissure in ano, which is safe, cost effective and reversible without significant complication. It has the potential of being used as a first line treatment in chronic fissure in ano.

The human imperative of stabilizing global climate change at 1.5°C.

Increased concentrations of atmospheric greenhouse gases have led to a global mean surface temperature 1.0°C higher than during the pre-industrial period. We expand on the recent IPCC Special Report on global warming of 1.5°C and review the additional risks associated with higher levels of warming, each having major implications for multiple geographies, climates, and ecosystems. Limiting warming to 1.5°C rather than 2.0°C would be required to maintain substantial proportions of ecosystems and would have clear benefits for human health and economies. These conclusions are relevant for people everywhere, particularly in low- and middle-income countries, where the escalation of climate-related risks may prevent the achievement of the United Nations Sustainable Development Goals.

Effect of insulin administration on ovarian function and estrus induction in acyclic goats.

The present experiment was conducted to study the effect of insulin on follicular development, ovarian steroid profiles and estrus induction in 14 non-descript acyclic goats. Experimental animals were randomly divided into two groups, i.e. group A (control, n=7) and group B (insulin-treated, n=7). The animals of treatment group were injected with long acting bovine insulin subcutaneously at 0.2IU/kg body weight once daily for five consecutive days. However animals of control group were injected with normal saline only as a placebo. Animals were subjected to detection of estrus twice daily (morning and evening) using an apronized buck and visual observation of estrus signs. Simultaneously animals of both groups were subjected to B-mode transrectal ovarian ultrasonsography for follicular development and blood collection on days 0, 4, 8, 12, 16, 20, 24 and 28 for the estimation of serum estradiol-17beta, progesterone and nitric oxide (NO) profiles, respectively. The results revealed significantly greater total number of follicles in insulin-treated goats as compared to control. The diameter of corpus luteum (CL) in insulin-administered goat was ranging between 5.14 and 6.14mm. Significantly higher concentrations of estradiol-17beta on days 4 and 8 and progesterone on days 12, 16, 20 and 24 were observed in insulin-treated goats as compared to control. Significantly higher concentration of NO was observed in insulin-treated group as compared to control. In the treatment group estrus was recorded in 5/7 goats (71%), with estrus duration of 19+/-2.6h at a mean interval of 189+/-17.7h from the initiation of treatment. Ovulation occurred in 4/5 (80%) of responding animals, as confirmed by detecting CL ultrasonographically. However in control none of the animal displayed signs of estrus during the observation period. The results indicate favorable effect of insulin on ovarian function.

Fluid and Medication Considerations in the Traumatized Patient.

This article reviews fluid therapy and medications in pediatric trauma. For resuscitation in the setting of hemorrhagic shock, isotonic crystalloid solution is the first-line agent of choice. Colloid solutions offer no additional benefit, introduce possible increased risks and cost more than crystalloids. Blood products, starting with pRBCs, should be introduced after 20-40 ml/kg of crystalloid has been administered if there is ongoing need for volume replacement. The use of a massive transfusion protocol of 1:1:1 (if >30 kg) or 30:20:20 (if <30 kg) of pRBCs:FFP:platelets is suggested after an initial 30 ml/kg of pRBcs has been administered. Cryoprecipitate should be given for documented low fibrinogen or ongoing bleeding after administration of 1 round of all 3 blood components. For patients at risk of massive hemorrhage, early administration of tranexamic acid with an initial loading dose of 15 mg/kg (maximum 1 g) is recommended. Choice of medication for intubation of the patient with Traumatic Brain Injury (TBI) may best be guided by physiology: in the TBI patient with a high mean arterial pressure, premedication with lidocaine, fentanyl and use of etomidate may be most appropriate, whereas in the hemodynamically compromised patient, use of ketamine alone may be considered. If needed, norepinephrine has been recommended as a temporizing agent for vasopressor support in the setting of fluid-refractory shock. Although controversial, in the setting of significant spinal cord injury, the potential benefits of administering 24-48 hours of steroids (initial 30 mg/kg of methylprednisolone within 8 hours of injury) may outweigh the risks especially in previously healthy pediatric patients.

Imaging during percutaneous coronary intervention for optimizing outcomes.

Angiography is the current gold standard for imaging during percutaneous coronary interventions but has significant limitations. Catheter-based intravascular imaging techniques such as intravascular ultrasound and the more recent optical coherence tomography have the potential to overcome these limitations and thus optimize clinical outcomes. In this update, we discussed the current applications of the available imaging techniques, existing evidence, continuing unmet needs, and potential areas for further research.

Ostial left main coronary artery chronic total occlusion presenting as chronic stable angina.

Significant left main coronary artery (LMCA) disease is found in 5-6% of all patients undergoing coronary angiography. It usually presents as acute coronary syndrome and is commonly associated with multi-vessel coronary artery disease (CAD). Complete occlusion of LMCA is a much rarer finding, since these patients usually present as unstable angina, myocardial infarction and cardiogenic shock. We report a case of a young female, who presented with chronic stable angina and had an isolated chronic total occlusion (CTO) of LMCA with no lesions in the other coronary arteries. Aortogram failed to demonstrate the stump of occluded LMCA and demonstrated the filling of the left coronary system from the right coronary artery. Apart from dyslipidemia, she had no other risk factors for CAD. She was extensively evaluated for non-atherosclerotic causes of LMCA CTO including vasculitis. She underwent coronary artery bypass graft successfully without any peri-procedural complications.

Minimizing Hepatic Artery Thrombosis and Establishing Safety of Grafts With Dual Arteries in Living Donor Liver Transplantation.

BACKGROUND: Hepatic artery thrombosis (HAT) is a dreaded complication following living donor liver transplantation (LDLT) and can lead to graft failure and biliary complications. We evaluated the results of our arterial anastomotic technique and outcomes in grafts with dual arterial supply. PATIENTS AND METHODS: Between July 2010 and June 2015, 225 patients underwent LDLT. The hepatic artery anastomosis was done using our "W technique". In grafts with a dual arterial supply, two anastomoses were performed unless there was significant pulsatile back-bleeding in the smaller artery after the larger anastomosis. RESULTS: The mean age of the recipients was 43±15.2 years (6 months to 66 years). There were 184 right liver, 30 left liver, 10 left lateral segment, and 1 dual lobe (right liver and left lateral segment) grafts. Twenty-three (10.2%) patients had 2 graft arteries, 10 of which required 2 separate anastomoses, and an interposition saphenous vein conduit was used in one. HAT occurred in 3 (1.3%) patients. The median intensive care unit and postoperative hospital stays were 5 and 14 days, respectively. Post-transplant operative mortality was 12.4%. There was no difference in mortality (8.7% vs 12.4%, P = >.99) and biliary complications (11.9% vs 21.7%, P = .19) between recipients of grafts with single or dual graft arteries, respectively. CONCLUSIONS: A careful surgical "W technique" and intraoperative confirmation of a good arterial flow helps in reducing the incidence of early HAT. The presence of two arteries in the graft was not associated with increased incidence of HAT, mortality, or biliary complications.