Bayesian approach for assessment of co-variances and genetic parameters of production and reproduction traits in Sahiwal cattle.

Precise as well as unbiased estimates of genetic parameters of economically important traits is important for framing breeding programmes meant for genetic progress of dairy animals. Present work was carried out to assess the co-variances along with genetic parameters of production and reproduction traits in Sahiwal cattle with Bayesian approach. Records of Sahiwal cattle kept at ICAR-National Dairy Research Institute, Karnal, Haryana for the period 1988-2016 were used. Heritability estimates ± posterior standard deviation for 305 days' milk yield (305DMY), lactation milk yield (LMY), lactation length (LL), peak yield (PY), days to attain peak yield (DPY), dry period (DP), gestation period (GP), calving to first insemination interval (CFI), days open (DO) and calving interval (CI) as obtained were 0.23 ± 0.03, 0.26 ± 0.03, 0.18 ± 0.03, 0.18 ± 0.03, 0.07 ± 0.02, 0.08 ± 0.02, 0.08 ± 0.02, 0.07 ± 0.02, 0.09 ± 0.02 and 0.02 ± 0.02, respectively and estimates of repeatability for the corresponding traits were 0.23 ± 0.03, 0.26 ± 0.03, 0.21 ± 0.03, 0.21 ± 0.03, 0.12 ± 0.02, 0.14 ± 0.03, 0.16 ± 0.03, 0.16 ± 0.03, 0.17 ± 0.03 and 0.17 ± 0.03. Moderate heritability values for 305DMY and LMY implies that sufficient additive genetic variability is available to obtain selection response. Repeatability estimates for 305DMY, LMY and PY were moderate, pointing towards the possibility of early appraisal of Sahiwal cattle for selection. Antagonistic genetic associations were observed between reproduction and production traits suggesting that reproduction traits need due attention while selecting high producing Sahiwal animals.

Development of sustainable performance index based on production, reproduction, health and longevity for genetic improvement of Sahiwal cattle.

An attempt was made for the first time in India to develop the performance indices for quantifying the relative emphasis to be given to production, reproduction, health and longevity traits for selection and genetic improvement of Sahiwal cattle in a sustainable manner. The study was conducted using information related to various production, reproduction, health and longevity traits spread over a period of 29 years in Sahiwal cattle. Using income and expenditure method, the relative economic values for 305 days milk yield (305DMY), average daily milk yield (ADMY), calving to first insemination interval (CFI), days open (DO) and longevity (LNG) were estimated as 1, 1, -6.62, -6.62 and 5.96 in Sahiwal cattle. A total of three performance indices were constructed using three-trait combination, that is production, reproduction and longevity before and after excluding the days suffered by the animals. Correlation between the aggregate genotype and index (RIH ) was computed to determine the accuracy of each performance index, and based on highest RIH value, best performance index was identified for selection of Sahiwal cattle. Developed indices were further assessed for robustness by increasing the relative economic values of the traits by 25% and 50%. The performance index (305DMY, DO and LNG) was found to be the best index before as well as after excluding days suffered by the animals, but in later case emphasis to be given to different traits was found to be more balanced. The best index indicated that about 47, 42 and 11% relative emphasis to be assigned to 305 days milk yield, days open and longevity for selection of Sahiwal animals for sustainable genetic improvement.

Point prevalence & risk factor assessment for hospital-acquired infections in a tertiary care hospital in Pune, India.

BACKGROUND & OBJECTIVES: Hospital-acquired infections (HAIs) are a major challenge to patient safety and have serious public health implications by changing the quality of life of patients and sometimes causing disability or even death. The true burden of HAI remains unknown, particularly in developing countries. The objective of this study was to estimate point prevalence of HAI and study the associated risk factors in a tertiary care hospital in Pune, India. METHODS: A series of four cross-sectional point prevalence surveys were carried out between March and August 2014. Data of each patient admitted were collected using a structured data entry form. Centers for Disease Control and Prevention guidelines were used to identify and diagnose patients with HAI. RESULTS: Overall prevalence of HAI was 3.76 per cent. Surgical Intensive Care Unit (ICU) (25%), medical ICU (20%), burns ward (20%) and paediatric ward (12.17%) were identified to have significant association with HAI. Prolonged hospital stay [odds ratio (OR=2.81), mechanical ventilation (OR=18.57), use of urinary catheter (OR=7.89) and exposure to central air-conditioning (OR=8.59) had higher odds of acquiring HAI (P<0.05). INTERPRETATION & CONCLUSIONS: HAI prevalence showed a progressive reduction over successive rounds of survey. Conscious effort needs to be taken by all concerned to reduce the duration of hospital stay. Use of medical devices should be minimized and used judiciously. Healthcare infection control should be a priority of every healthcare provider. Such surveys should be done in different healthcare settings to plan a response to reducing HAI.

Influence of Temperature and Humidity on Pregnancy Rate of Murrah Buffaloes under Subtropical Climate.

Heat stress has adverse effects on fertility of dairy animals. Decline in fertility is linearly associated with an increase in combination of both temperature and humidity. The purpose of this study was to investigate the relationship between temperature humidity index (THI) and the pregnancy rate of Murrah buffaloes in a subtropical climate. The effects of genetic and non-genetic factors viz., sire, parity, period of calving and age group at first calving were found non-significant on pregnancy rate. The effect of THI was found significant (p<0.001) on pregnancy rate of Murrah buffaloes calved for first time and overall pregnancy rate. The threshold THI affecting the pregnancy rate was identified as THI 75. The months from October to March showed THI<75 and considered as non heat stress zone (NHSZ), while months from April to September were determined as heat stress zone (HSZ) with THI≥75. The lowest overall pregnancy rate (0.25) was obtained in July with THI 80.9, while the highest overall pregnancy rate (0.59) was found in November with THI 66.1. May and June were identified as critical heat stress zone (CHSZ) within the HSZ with maximum decline (-7%) in pregnancy rate with per unit increase in THI. The highest overall pregnancy rate was estimated as 0.45 in NHSZ with THI value 56.7 to 73.2. The pregnancy rate was found to have declined to 0.28 in HSZ with THI 73.5 to 83.7. However, the lowest pregnancy rate was estimated as 0.27 in CHSZ with THI value 80.3 to 81.6.

Lactoferrin gene promoter variants and their association with clinical and subclinical mastitis in indigenous and crossbred cattle.

Lactoferrin (Lf) gene promoter was screened for the presence of single nucleotide polymphism in indigenous and crossbred cattle from North India and to evaluate its association with Mastitis. Study revealed the presence of genetic variation in regulatory region of bovine Lactoferrin gene using PCR-RFLP technique. Three genotypes namely GG, GH and HH were identified. A single nucleotide change, from guanine to adenine at 25th position was found to be significantly associated (p<0.05) with clinical mastitis in indigenous Sahiwal and crossbred Karan Fries cattle maintained at organised herd of National Dairy Research Institute, Karnal. A non-significant association was observed between subclinical mastitis, somatic cell score (SCS), and GG genotype in Karan Fries cattle, however, a lower SCS was observed in animals having GG genotype. Overall a lower incidence of clinical mastitis was recorded in those animals having GG genotype of Lf in Sahiwal and Karan Fries (KF) cattle. The SNP identified in the promoter region may effect expression lactoferrin protein, which may lead to different levels of antibacterial and anti-inflammatory activity of Lf gene. Results from this study indicated the probable role played by Lactoferrin promoter to serve as candidate gene for mastitis susceptibility among indigenous and crossbred milch cattle.

ABC-VED analysis of expendable medical stores at a tertiary care hospital.

BACKGROUND: The modern system of medicine has evolved into a complex, sophisticated and expensive treatment modality in terms of cost of medicines and consumables. In any hospital, approximately 33% of total annual budget is spent on buying materials and supplies including medicines. ABC (Always, Better Control)-VED (Vital, Essential, Desirable) analysis of medical stores of a large teaching, tertiary care hospital of the Armed Forces was carried out to identify the categories of drugs needing focused managerial control. METHODS: Annual consumption and expenditure data of expendable medical stores for one year was extracted from the drug expense book, followed by classification on its annual usage value. Subsequently, the factor of criticality was applied to arrive at a decision matrix for understanding the need for selective managerial control. RESULTS: The study revealed that out of 1536 items considered for the study, 6.77% (104), 19.27% (296) and 73.95% (1136) items were found to be A, B and C category items respectively. VED analysis revealed that vital items (V) accounted for 13.14% (201), essential items (E) for 56.37% (866) and desirable accounted for 30.49% items (469). ABC-VED matrix analysis of the inventory reveals that only 322 (21%) items out of an inventory of 1536 drugs belonging to category I will require maximum attention. CONCLUSION: Scientific inventory management tools need to be applied routinely for efficient management of medical stores, as it contributes to judicious use of limited resources and resultant improvement in patient care.

Applications of pathology-assisted image analysis of immunohistochemistry-based biomarkers in oncology.

Immunohistochemistry-based biomarkers are commonly used to understand target inhibition in key cancer pathways in preclinical models and clinical studies. Automated slide-scanning and advanced high-throughput image analysis software technologies have evolved into a routine methodology for quantitative analysis of immunohistochemistry-based biomarkers. Alongside the traditional pathology H-score based on physical slides, the pathology world is welcoming digital pathology and advanced quantitative image analysis, which have enabled tissue- and cellular-level analysis. An automated workflow was implemented that includes automated staining, slide-scanning, and image analysis methodologies to explore biomarkers involved in 2 cancer targets: Aurora A and NEDD8-activating enzyme (NAE). The 2 workflows highlight the evolution of our immunohistochemistry laboratory and the different needs and requirements of each biological assay. Skin biopsies obtained from MLN8237 (Aurora A inhibitor) phase 1 clinical trials were evaluated for mitotic and apoptotic index, while mitotic index and defects in chromosome alignment and spindles were assessed in tumor biopsies to demonstrate Aurora A inhibition. Additionally, in both preclinical xenograft models and an acute myeloid leukemia phase 1 trial of the NAE inhibitor MLN4924, development of a novel image algorithm enabled measurement of downstream pathway modulation upon NAE inhibition. In the highlighted studies, developing a biomarker strategy based on automated image analysis solutions enabled project teams to confirm target and pathway inhibition and understand downstream outcomes of target inhibition with increased throughput and quantitative accuracy. These case studies demonstrate a strategy that combines a pathologist's expertise with automated image analysis to support oncology drug discovery and development programs.

Novel high throughput screen reports that benzo(a)pyrene overrides mouse trophoblast stem cell multipotency, inducing SAPK activity, HAND1 and differentiated trophoblast giant cells.

INTRODUCTION: Cultured mouse trophoblast stem cells (mTSC) maintain proliferation/normal stemness (NS) under FGF4, which when removed, causes normal differentiation (ND). Hypoxic, or hyperosmotic stress forces trophoblast giant cells (TGC) differentiate. Hypoxic, hyperosmotic, and genotoxic benzo(a)pyrene (BaP), which is found in tobacco smoke, force down-regulation of inhibitor of differentiation (Id)2, enabling TGC differentiation. Hypoxic and hyperosmotic stress induce TGC by SAPK-dependent HAND1 increase. Here we test whether BaP forces mTSC-to-TGC while inducing SAPK and HAND1. METHODS: Hand1 and SAPK activity were assayed by immunoblot, mTSC-to-TGC growth and differentiation were assayed at Tfinal after 72hr exposure of BaP, NS, ND, Retinoic acid (RA), or sorbitol. Nuclear-stained cells were micrographed automatically by a live imager, and assayed by ImageJ/FIJI, Biotek Gen 5, AIVIA proprietary artificial intelligence (AI) software or open source, CellPose artificial intelligence/AI software. RESULTS: BaP (0.05-1µM) activated SAPK and HAND1 without diminishing growth. TSC-to-TGC differentiation was assayed with increasingly accuracy for 2-4 N cycling nuclei and >4 N differentiating TGC nuclei, using ImageJ/FIJI, Gen 5, AIVIA, or CellPose AI software. The AIVIA and Cellpose AI software matches human accuracy. The lowest BaP effects on SAPK activation/HAND1 increase are >10-fold more sensitive than similar effects for mESC. RA induces 44-47% 1st lineage TGC differentiation, but the same RA dose induces only 1% 1st lineage mESC differentiation. DISCUSSION: First, these pilot data suggest that mTSC can be used in high throughput screens (HTS) to predict toxicant exposures that force TGC differentiation. Second, mTSC differentiated more cells than mESC for similar stress exposures, Third, open source AI can replace human micrograph quantitation and enable a miscarriage-predicting HTS.

Lectin-driven maturation of cytotoxic effector cells: the nature of effector memory.

In an attempt to define further the activation of effector T-cell function with concanavalin A (Con A), we examined the ability of Con A to generate effector memory in mouse lymph node cells in vitro. In the course of these studies, it became necessary to define parameters by which memory could be defined. These parameters include length of time of exposure to signal required to generate full cytotoxic function; time of onset and kinetics of blast cell formation; requirement for DNA synthesis; sensitivity to the drug 5'-bromodeoxyuridine; and kinetics of the cytotoxic response to alloantigen. By these criteria, mouse lymph node cells exposed 12 days previously to Con A behave qualitatively differently from unprimed mouse lymphocytes. We found that the time of onset and kinetics of DNA synthesis could not be used to distinguish primary and secondary cytotoxic responses. We propose that the parameters defined in these stidues can be applied generally in determining whether a given cytotoxic response involves primed (memory) or unprimed cells.

Sporadic hemiplegic migraine in children: a report of two new cases.

Two cases of sporadic hemiplegic migraine, which fulfilled the diagnostic criteria as laid down in International Classification of Headache Disorders (ICHD)-2, are reported in children. In the first case, two unusual features were noted, namely, the occurrence of dysphsia in association with a left hemiparesis and the spread of sensory symptoms to the contralateral side during attacks. The second case is perhaps the youngest patient reported with this disorder.

Migraine, epileptic seizures and psychogenic non-epileptic seizures: observations in Indian patients in a clinic-based study.

The present study focuses on the incidence of epileptic seizures in 1000 patients (200 children and 800 adults) with migraine. Very few patients with migraine had history of epileptic seizures. No patient had migraine-induced seizures and none had seizure-induced migraine like headaches. Occurrence of psychogenic non-epileptic attacks during migraine headaches had not been highlighted in the past. In the present study, about a quarter of patients with migraine, especially adolescent and adult females, had history of psychogenic non-epileptic seizures during attacks of acute migraine. This appears to be the first report on these features in Indian subjects with migraine.

Mutations in growth hormone gene affect stability of protein structure leading to reduced growth, reproduction, and milk production in crossbred cattle-an insight.

The GH has a multifaceted role in growth, reproduction, and milk production. Nonsynonymous mutations identified as L153V were observed from GH1 (L) to GH2 (V) with higher genotypic frequency for GH1 being 0.87. GH2 (V) allele was identified as a rare allele and the population followed Hardy-Weinberg equilibrium. LL homozygote variant had significantly better growth, reproduction, and expected milk production at different ages in crossbred (CB) males. Reports are scanty explaining the molecular mechanism of how individuals with LV genotype were phenotypically inferior to that of wild-type LL. In the present study, it was explored that GH peptide with LV heterozygotes of GH gene, were observed to have reduced structural stability thermodynamically and thus functionally leads to reduced economic traits in CB animals. The fact was first time reported and confirmed through genomic analysis, bioinformatics, and later confirmed through immunohistochemistry. Differential expression analysis of the GH gene with respect to other genes in the hypothalamus-pituitary growth axis of CB cattle was also studied to have a complete insight of the GH gene.

Cost-effectiveness Analysis for Technology Acquisition.

BACKGROUND: In a developing country with limited resources, it is important to utilize the total cost visibility approach over the entire life-cycle of the technology and then analyse alternative options for acquiring technology. METHODS: The present study analysed cost-effectiveness of an "In-house" magnetic resonance imaging (MRI) scan facility of a large service hospital against outsourcing possibilities. Cost per unit scan was calculated by operating costing method and break-even volume was calculated. Then life-cycle cost analysis was performed to enable total cost visibility of the MRI scan in both "In-house" and "outsourcing of facility" configuration. Finally, cost-effectiveness analysis was performed to identify the more acceptable decision option. RESULT: Total cost for performing unit MRI scan was found to be Rs 3,875 for scans without contrast and Rs 4,129 with contrast. On life-cycle cost analysis, net present value (NPV) of the "In-house" configuration was found to be Rs-(4,09,06,265) while that of "outsourcing of facility" configuration was Rs-(5,70,23,315). Subsequently, cost-effectiveness analysis across eight Figures of Merit showed the "In-house" facility to be the more acceptable option for the system. CONCLUSION: Every decision for acquiring high-end technology must be subjected to life-cycle cost analysis.

Observations on juvenile myoclonic epilepsy amongst ethnic Bengalees in West Bengal--an Eastern Indian State.

BACKGROUND: Juvenile myoclonic epilepsy (JME) is not too uncommonly encountered in Indian neurological practice. A number of reports from different parts of India have documented the clinical phenomenology and EEG characteristics of this genetically determined epileptic syndrome. However, no study has yet been reported from the Eastern part of India and none done so far in patients in a specific ethnic group. Furthermore therapy response and follow up data are not available in detail in the Indian studies. OBJECTIVE: To study disease expression, EEG characteristics and therapy response of JME patients in ethnic Bengalees in West Bengal, an Eastern Indian State, in a clinic based study. MATERIAL AND METHODS: 200 patients with JME attending the Neurology Department of the Institute have been followed up for 5 years and different parameters of disease expression as outlined above have been studied. RESULTS: Overall clinical disease expression has been found to be similar in this clinic based study in ethnic Bengalees as compared to other reports from India and elsewhere. About 16% of patients showed a relative resistance to Valproate therapy. Hundred percent of patients in whom therapy withdrawal was attempted, relapsed within<1-2 years. Amongst female patients (132), 16 developed features of polycystic ovarian syndrome while on Valproate therapy. In over half of them, the symptoms regressed after successful switch over from Valproate to Clobazam. 12/132 female patients became pregnant during follow up and while on Valproate; teratogenic effect was evident in only one such patient. CONCLUSIONS: Phenotypic variations in disease expression including therapy response have been noted within a single ethnic group of patients attending the clinic and might account for genetic heterogeneity noted in molecular genetic studies. JME cannot really be called a very 'benign' epileptic syndrome; recurrence after therapy withdrawal almost invariably occurs.

CNS and anticonvulsant activity of a non-protein toxin (KC-MMTx) isolated from King Cobra (Ophiophagus hannah) venom.

In the present study, King Cobra (Ophiophagus hannah) venom was subjected to TLC followed by column chromatography/HPLC to isolate and purify a non-protein toxin designated as KC-MMTx. (1)H NMR, IR and EIMS studies showed KC-MMTx likely to be a 282 D unsaturated aliphatic acid having molecular formula C18H34O2. The minimum lethal dose of KC-MMTx was 200 microg/kg (i.v.) and 350 microg/kg (i.p.) in Swiss albino male mice. It significantly increased pentobarbitone induced sleeping time and significantly decreased the body temperature of male albino mice. It provided protection against amphetamine aggregate toxicity in mice but failed to protect amphetamine stereotypy in male albino rats. KC-MMTx provided significant protection against drug (strychnine, pentylenetetrazole, yohimbine) induced convulsions in male albino mice. It increased serum Na+ and decreased serum Ca2+ significantly in male mice. MAO activity and brain neurotransmitter levels in male mice were altered significantly. Further detailed study is warranted on the CNS, anticonvulsant potential of KC-MMTx, which may lead to the development of newer therapeutic tools in the near future.

Daboia russellii and Naja kaouthia venom neutralization by lupeol acetate isolated from the root extract of Indian sarsaparilla Hemidesmus indicus R.Br.

The present study reports the isolation and purification of lupeol acetate from the methanolic root extract of Indian medicinal plant Hemidesmus indicus (L.) R.Br. (family: Asclepiadaceae) which could neutralize venom induced action of Daboia russellii and Naja kaouthia on experimental animals. Lupeol acetate could significantly neutralize lethality, haemorrhage, defibrinogenation, edema, PLA(2) activity induced by Daboia russellii venom. It also neutralized Naja kaouthia venom induced lethality, cardiotoxicity, neurotoxicity and respiratory changes in experimental animals. Lupeol acetate potentiated the protection by snake venom antiserum action against Daboia russellii venom induced lethality in male albino mice. Venom induced changes in lipid peroxidation and super oxide dismutase activity was antagonized by lupeol acetate. Snake venom neutralization by lupeol acetate and its possible mechanism of action has been discussed.

Occurrence of non-protein low molecular weight cardiotoxin in Indian King Cobra (Ophiophagus hannah) Cantor 1836, venom.

Pathophysiology due to snakebite is a combined effect of various actions of the complex venom constituents. Importance of protein toxins in snake envenomation is well known. The present investigation reports the existence of nonprotein/nonpetide low molecular weight toxin in Indian King Cobra venom, which plays an important role in envenomation consequences in experimental animal models. A group of non-peptidic toxins (OH-NPT1) was isolated from Indian King Cobra Ophiophagus hannah by thin layer chromatography and silica gel column chromatography. UV, IR, NMR and (ESI) TOF-MS studies characterized the OH-NPT1 as a mixture of aliphatic acids having molecular weights 256, 326 and 340Da. The minimum lethal dose of OH-NPT1 was found to be 2.5 microg/20g (iv) and 4microg/20g (ip) in male albino mice. The cardiotoxic property of OH-NPT1 was established through studies on isolated guinea pig heart and auricle preparations, ECG studies in albino rat and estimation of LDH1/LDH and CPK-MB/CPK ratio in Swiss albino mice. Commercial antiserum failed to neutralize the lethality and cardiotoxicity of the toxin. However, calcium and magnesium effectively neutralized the lethal action.

Evaluation of fertility in relation to milk production and productivity of Murrah buffaloes.

Intense selection of buffaloes for milk production at organized herds of the country without giving due attention to fertility traits has lead to deterioration in their performances. The study was initiated to assess the relationship between milk production and productivity with fertility in Murrah buffaloes. In the present study, fertility was defined in terms of pregnancy rate of Murrah buffaloes. Pregnancy rate measures the percentage of non-pregnant animals that become pregnant during each oestrous cycle. Data pertaining to 1224 lactation records of Murrah buffaloes spread over a period 22 years from 1993 to 2014 were analyzed in the study. It was observed that pregnancy rate is negatively associated with 305days or less milk yield (-0.08±0.04) and wet average (-0.12±0.02) and positively associated with life time (0.15±0.03) in Murrah buffaloes. Wet average is defined as average daily milk yield per lactation of Murrah buffaloes. To achieve around 2000kg 305days or less milk yield and 7.5kg wet average, the level of pregnancy rate in Murrah buffaloes should vary between 30 and 50%. The per unit change in pregnancy rate with respect to milk yield in Murrah buffaloes, were studied using regression analysis. The results suggest that increasing hundred kilogram in 305days or one kilogram milk yield per day in 305days, pregnancy rate reduced by 0.9% in overall lactations of Murrah buffaloes. By increasing hundred kilogram life time 305days or less milk yield and one kilogram life time wet average, pregnancy rate of Murrah buffaloes reduced by about 0.2% and 0.34%, respectively. The study quantifies the decline of pregnancy rate with increase of lactation milk yield and wet average in Murrah buffaloes and emphasis the importance of fertility i.e. pregnancy rate in the evaluation and breeding programmes of Murrah buffaloes.

Distribution of Class II integrons and their contribution to antibiotic resistance within Enterobacteriaceae family in India.

BACKGROUND: Integrons are the main contributors to the development of multidrug resistance (MDR) among Gram-negative bacilli. There is a lack of knowledge about the molecular relation between gene cassettes and antibiotic resistance in India. OBJECTIVE: In this study, we have investigated the occurrence of Class II integron and their cassette array among Enterobacteriaceae. MATERIALS AND METHODS: A total of 268 MDR non-duplicate strains of Enterobacteriaceae were collected from Silchar Medical College and Hospital, Silchar, Assam, India, during June 2012 to May 2013. Polymerase chain reaction was performed for detection of the integrase genes and gene cassettes within the Class II integron which were further analysed by sequencing. RESULTS: Class II integron was observed in 47 isolates. Four different gene cassette arrangements were detected: dfrA1-sat2-aadA1; dfrA1-sat2-aadA1-orfX-ybeA-ybfA-ybfB-ybgA; dfrA12-sat2-aadA1; and dfrA1-linF-aadA1. The most prevalent cassette combination was dfrA1-sat2-aadA1. This study has also identified a set of gene cassette associated with linF gene instead of sat2 gene. CONCLUSION: Further investigation is required to determine the current situation and important reservoir of Class II integron for the transmission of drug resistance among Enterobacteriaceae and their contribution to antimicrobial resistance in hospital environment .