Influence of Bacterial Contamination and Antibiotic Sensitivity on Cryopreserved Sperm Quality of Indian Red Jungle Fowl.

Aims: Bacterial contamination may occur in feces during collection and processing of semen. Bacteria not only compete for nutrients with spermatozoa but also produce toxic metabolites and endotoxins and affect sperm quality. The aim of the present study was to investigate the effect of antibiotic supplementation on the sperm quality of Indian red jungle fowl, estimation and isolation of bacterial species and their antibiotic sensitivity. Materials and Methods: Semen was collected and initially evaluated, diluted, and divided into six experimental extenders containing gentamicin (2.5 µg/mL), kanamycin (31.2 µg/mL), neomycin (62.5 mg/mL), penicillin (200 U/mL), and streptomycin (250 µg/mL), and a control having no antibiotics were cryopreserved and semen quality was evaluated at post-dilution, post-cooling, post-equilibration, and post-thawing stages (Experiment 1). A total aerobic bacterial count was carried out after culturing bacteria (Experiment 2) and subcultured for antibiotic sensitivity (Experiment 3). Results: It was shown that penicillin-containing extender improved semen quality (sperm motility, plasma membrane integrity, viability, and acrosomal integrity) compared with the control and other extenders having antibiotics. The bacteria isolated from semen were Escherichia coli, Staphylococcus spp., and Bacillus spp. Antibiotic sensitivity results revealed that E. coli shows high sensitivity toward neomycin, kanamycin, and penicillin. Staphylococcus spp. shows high sensitivity toward streptomycin, neomycin, and penicillin. Bacillus spp. shows high sensitivity toward kanamycin and penicillin. Conclusions: It was concluded that antibiotics added to semen extender did not cause any toxicity and maintained semen quality as that of untreated control samples, and penicillin was identified as most effective antibiotic. It is recommended that penicillin can be added to the semen extender for control of bacterial contamination without affecting the semen quality of Indian red jungle fowl.

Association of Visfatin gene polymorphism with obesity related metabolic disorders among Pakistani population: a case control study.

In recent years, the global prevalence of obesity and its associated metabolic disorders has reached alarming levels, presenting a significant challenge to public health worldwide. Visfatin, also known as pre-B cell colony-enhancing factor (PBEF) or nicotinamide phosphoribosyltransferase (NAMPT), is an adipokine that has been implicated in various physiological processes, including glucose homeostasis, lipid metabolism, and inflammation. The main objective of this proposed study is to find out the association between visfatin genetic variants and metabolic syndrome. The sample size of the study consisted of 300 blood samples (150 control and 150 cases). This study found that the genotypic frequency of visfatin SNPs, including rs2302559 (OD: 18.222; 95% CI 10.228-32.466; p-value < 0.001) and rs1215113036 (OD: 129.40; 95% CI 44.576-375.693; p-value < 0.001) were significantly associated with metabolic syndrome. Moreover, the frequency of the mutant alleles of both visfatin SNPs was found to be higher in patients with metabolic syndrome as compared to controls. Results of the current study indicate that people with any genetic variation of Visfatin, such as rs2302559 and rs1215113036, are more likely to develop metabolic syndrome. Visfatin genetic variants are linked to an increased risk of metabolic syndrome, implying it's role in disease pathophysiology.

Dietary Patterns and Physical Activity Levels Among People With Type 2 Diabetes.

Objective: The aim of our study was to assess the association of eating habits with the dietary patterns of people with diabetes. Methods: This cross-sectional study was conducted at National Institute of Diabetes and Endocrinology (NIDE), Dow University Hospital in Karachi, Pakistan. A total 301 patients aged >18 with type 2 diabetes came to Outpatient department were recruited. Structured questionnaire was used to assess general characteristics, Anthropometric measurements, biochemical markers, and dietary intake. Results: A total of 301 patients with type 2 diabetes mellitus were included in this study. The average age of patients was 51.6 (SD ± 11.1) years which ranged from 21 to 80 years whereas the average BMI was 27.2 kg / m2 (SD ± 5.6). Overall, 42% of patients were found to often have less than 1 serving of fruit, and 45% had less than 3 servings of vegetables daily. Of all, 77 (26%) patients often distributed carbohydrates all over the day. Results revealed that HbA1c was higher in those patients who took more than 3 roti (approximately each roti weight 60-80 g) (P-value < .001) and full plate rice approximately 300 to 355 g cooked weight in a whole day (P-value < .001) as compared to those patients whose intake of roti was 3 or less than 3 and rice was a half plate. Moreover, out of 301 patients, 102 were found physically active (52% females and 48% males), while 199 were not active. Pain in legs and lack of motivation were common barrier to physical activity. Conclusion: Our study revealed that patients' fruits and vegetable intake was not optimum, diet was not balanced and the quantity of starchy carbohydrates was not controlled which may affect their HBA1C levels. Proper counseling and awareness about the importance of a balanced diet and portion control in diabetes are needed.

Prevalence and Distribution of Hard Ticks and Their Associated Risk Factors in Sheep and Goats from Four Agro-Climatic Zones of Khyber Pakhtunkhwa (KPK), Pakistan.

Hard ticks are blood-sucking ectoparasites infesting all types of domestic ruminants throughout the world and serve as vectors in the dissemination of a wide variety of pathogens. Sheep and goat farming is a vital economic source for resource-poor farming communities in the Khyber Pakhtunkhwa (KPK) province of Pakistan. Aim: The aim of present study is to study the epidemiological profile of ticks in various agro-climatic zones of KPK. Materials and Methods: A total of 1500 (882 sheep and 618 goats) of all age groups and sexes were examined for the presence of ticks belonging to six districts in four agro-climatic zones of KPK province, Pakistan. Non-probability sampling was undertaken based on animal hosts' selection, collection of hard ticks, and epidemiological parameters. Ticks collected from sheep and goats were identified phenotypically using standard keys. Results: The results revealed that Rhipicephalus microplus (36.2%) was the predominant tick species followed by Hyalomma anatolicum (25.2%), Hyalomma trancatum (18.1%), Hyalomma aegyptium (11.3%), Hyalomma asiaticum (6.9%), and Haemephysalis bispinosa (2.4%). Different epidemiological parameters showed that weather, humidity, and host age negatively affect (p < 0.05) tick load while temperatures (Minimum and Maximum), sunshine (hrs), and precipitation positively affected (p < 0.05) tick load. Host sex only showed a positive association with tick load (p > 0.05). The highest value of tick diversity index (H) 0.36748 was noted for Hyalomma anatolicum as this tick species was commonly distributed in three agro-climatic zones except in the Suleiman Piedmont zone, while Haemaphysalis bispinosa has the lowest diversity index (0.0334) as it was only found in the Central Valley Plains zone of KPK. The Shannon diversity index of tick diversity was highest at Barikot while the lowest index value was at Kabal (2.407). Similarly, a higher Simpson's diversity index was at Barikot (0.9554) and the lowest hard index was noted at Takht bai (2.874). The dominance index showed that the tick species were more dominant at Takht bai (0.1251), while the lowest dominance was at Barikot (0.04465). Conclusions: It has been concluded that tick diversity and distribution, abundance, reproduction, development, and survival depend on prevailing climatic conditions. The present study would not only help to identify the tick species but also facilitate in devising effective control measures to minimize the transmission pathogens in sheep and goats being reared in the various agro-climatic zones of KPK province, Pakistan.

Seasonal distribution and habitat use preference of Barking deer (Muntiacus vaginalis) in Murree-Kotli Sattian-Kahuta National Park, Punjab Pakistan / Distribuição sazonal e preferência de uso de habitat de veados-latidos (Muntiacus vaginalis) no Parque Nacional Murree-Kotli Sattian-Kahuta, Punjab Paquistão

Microhabitat factors associated with the habitat of barking deer (Muntiacus vaginalis) were examined and compared between summer and winter seasons. Habitat characteristics and preferred habitat were measured by locating direct and indirect signs. To quantify the habitat utilization of barking deer, each selected study site was sampled for floral diversity from 2015 to 2017. Quadrats were deployed along transect lines to determine seasonal distribution. Barking deer were not evenly distributed across vegetation types in the study area; they occurred more often in the broad-leaved forest than in Chir pine forest, at an elevational range of 550-850 m, in thick vegetation on steep slopes. The most preferred habitat included trees and shrubs with 30% and 69% cover, respectively. Barking deer avoided thicker tree cover, possibly as it hinders movement and escape from predators. No significant difference (χ2 = 6.37, df = 3, p = 0.19) in seasonal vegetation cover was recorded.

Fatores de micro-hábitat associados ao hábitat do veado (Muntiacus vaginalis) foram examinados e comparados entre as estações de verão e inverno. As características do hábitat e o hábitat preferido foram medidos, localizando sinais diretos e indiretos. Para quantificar a utilização do hábitat de cervos-latidos, cada local de estudo selecionado foi amostrado para a diversidade floral de 2015 a 2017. Quadrats foram implantados ao longo de linhas de transecto para determinar a distribuição sazonal., Veados-latidos não foram distribuídos uniformemente pelos tipos de vegetação na área de estudo; ocorreram com mais frequência na floresta de folhas largas do que na floresta de pinheiros Chir, em uma faixa de elevação de 550-850 m, em vegetação densa, em encostas íngremes. O hábitat mais preferido incluía árvores e arbustos com 30% e 69% de cobertura, respectivamente. O veado-latido evitou uma cobertura de árvores mais espessa, possivelmente porque impede o movimento e a fuga de predadores. Nenhuma diferença significativa (χ2 = 6,37; df = 3; p = 0,19) na cobertura vegetal sazonal foi registrada.

Seasonal distribution and habitat use preference of Barking deer (Muntiacus vaginalis) in Murree-Kotli Sattian-Kahuta National Park, Punjab Pakistan / Distribuição sazonal e preferência de uso de habitat de veados-latidos (Muntiacus vaginalis) no Parque Nacional Murree-Kotli Sattian-Kahuta, Punjab Paquistão

Microhabitat factors associated with the habitat of barking deer (Muntiacus vaginalis) were examined and compared between summer and winter seasons. Habitat characteristics and preferred habitat were measured by locating direct and indirect signs. To quantify the habitat utilization of barking deer, each selected study site was sampled for floral diversity from 2015 to 2017. Quadrats were deployed along transect lines to determine seasonal distribution. Barking deer were not evenly distributed across vegetation types in the study area; they occurred more often in the broad-leaved forest than in Chir pine forest, at an elevational range of 550-850 m, in thick vegetation on steep slopes. The most preferred habitat included trees and shrubs with 30% and 69% cover, respectively. Barking deer avoided thicker tree cover, possibly as it hinders movement and escape from predators. No significant difference (χ2 = 6.37, df = 3, p = 0.19) in seasonal vegetation cover was recorded.

Fatores de micro-hábitat associados ao hábitat do veado (Muntiacus vaginalis) foram examinados e comparados entre as estações de verão e inverno. As características do hábitat e o hábitat preferido foram medidos, localizando sinais diretos e indiretos. Para quantificar a utilização do hábitat de cervos-latidos, cada local de estudo selecionado foi amostrado para a diversidade floral de 2015 a 2017. Quadrats foram implantados ao longo de linhas de transecto para determinar a distribuição sazonal., Veados-latidos não foram distribuídos uniformemente pelos tipos de vegetação na área de estudo; ocorreram com mais frequência na floresta de folhas largas do que na floresta de pinheiros Chir, em uma faixa de elevação de 550-850 m, em vegetação densa, em encostas íngremes. O hábitat mais preferido incluía árvores e arbustos com 30% e 69% de cobertura, respectivamente. O veado-latido evitou uma cobertura de árvores mais espessa, possivelmente porque impede o movimento e a fuga de predadores. Nenhuma diferença significativa (χ2 = 6,37; df = 3; p = 0,19) na cobertura vegetal sazonal foi registrada.

Seasonal distribution and habitat use preference of Barking deer (Muntiacus vaginalis) in Murree-Kotli Sattian-Kahuta National Park, Punjab Pakistan

ABSTRACT Microhabitat factors associated with the habitat of barking deer (Muntiacus vaginalis) were examined and compared between summer and winter seasons. Habitat characteristics and preferred habitat were measured by locating direct and indirect signs. To quantify the habitat utilization of barking deer, each selected study site was sampled for floral diversity from 2015 to 2017. Quadrats were deployed along transect lines to determine seasonal distribution. Barking deer were not evenly distributed across vegetation types in the study area; they occurred more often in the broad-leaved forest than in Chir pine forest, at an elevational range of 550-850 m, in thick vegetation on steep slopes. The most preferred habitat included trees and shrubs with 30% and 69% cover, respectively. Barking deer avoided thicker tree cover, possibly as it hinders movement and escape from predators. No significant difference (2 = 6.37, df = 3, p = 0.19) in seasonal vegetation cover was recorded.

RESUMO Fatores de micro-hábitat associados ao hábitat do veado (Muntiacus vaginalis) foram examinados e comparados entre as estações de verão e inverno. As características do hábitat e o hábitat preferido foram medidos, localizando sinais diretos e indiretos. Para quantificar a utilização do hábitat de cervos-latidos, cada local de estudo selecionado foi amostrado para a diversidade floral de 2015 a 2017. Quadrats foram implantados ao longo de linhas de transecto para determinar a distribuição sazonal., Veados-latidos não foram distribuídos uniformemente pelos tipos de vegetação na área de estudo; ocorreram com mais frequência na floresta de folhas largas do que na floresta de pinheiros Chir, em uma faixa de elevação de 550-850 m, em vegetação densa, em encostas íngremes. O hábitat mais preferido incluía árvores e arbustos com 30% e 69% de cobertura, respectivamente. O veado-latido evitou uma cobertura de árvores mais espessa, possivelmente porque impede o movimento e a fuga de predadores. Nenhuma diferença significativa (2 = 6,37; df = 3; p = 0,19) na cobertura vegetal sazonal foi registrada.

Seasonal distribution and habitat use preference of Barking deer (Muntiacus vaginalis) in Murree-Kotli Sattian-Kahuta National Park, Punjab Pakistan.

Microhabitat factors associated with the habitat of barking deer (Muntiacus vaginalis) were examined and compared between summer and winter seasons. Habitat characteristics and preferred habitat were measured by locating direct and indirect signs. To quantify the habitat utilization of barking deer, each selected study site was sampled for floral diversity from 2015 to 2017. Quadrats were deployed along transect lines to determine seasonal distribution. Barking deer were not evenly distributed across vegetation types in the study area; they occurred more often in the broad-leaved forest than in Chir pine forest, at an elevational range of 550-850 m, in thick vegetation on steep slopes. The most preferred habitat included trees and shrubs with 30% and 69% cover, respectively. Barking deer avoided thicker tree cover, possibly as it hinders movement and escape from predators. No significant difference (χ2 = 6.37, df = 3, p = 0.19) in seasonal vegetation cover was recorded.

Role of nebulized epinephrine in moderate bronchiolitis: a quasi-randomized trial.

BACKGROUND/AIMS: Bronchiolitis is the most common lower respiratory illness that characteristically affects the children below 2 years of age accounting about 2-3% of patients admitted to hospital each year [1-4]. We compared the effect of racemic epinephrine (RE) and 3% hypertonic saline (HS) nebulization on the length of stay (LOS) in the hospital. METHODS: We looked at the infants with moderate bronchiolitis, from October 2013 to March 2014. Out of eighty cases, 16 in HS and 18 in RE groups were enrolled. At the time of admission, 0.2 ml of RE added to 1.8 ml of distilled water was nebulized to RE group, as compared with 2 ml of 3% HS in nebulized form. RE was re-administered if needed on 6 h in comparison with 3% HS at the frequency of 1 to 4 h. RESULTS: One infant from RE group and three infants from HS group were excluded due to progression towards severe bronchiolitis. The LOS in RE group ranged between 18 and 160 h (mean 45 h), while in HS group, LOS was 18.50-206 h (mean 74.3 h). The LOS was significantly short in RE group (p value 0.015) which was statistically significant. CONCLUSION: Racemic epinephrine nebulization as first-line medication may significantly reduce the length of hospital stay in infants with moderate bronchiolitis in comparison with nebulized HS.

Effect of Quercetin on Oxidative Stress, Mitochondrial Activity, and Quality of Indian Red Jungle Fowl (Gallus gallus murghi) Sperm.

Aim: The study was designed to elucidate the effects of quercetin in an extender on oxidative stress, mitochondrial activity and quality of Indian red jungle fowl (Gallus gallus murghi) sperm during cryopreservation. Materials and Methods: Semen was collected from seven adult males through abdominal massage and evaluated for semen volume, concentration, and motility. The qualifying semen ejaculates having >80% motility were diluted in red fowl extenders with 0 (control), 5, 10, 15, and 20 mM quercetin. Diluted semen was frozen following a glycerol-based protocol. Semen quality (motility, plasma membrane integrity, viability, acrosome integrity, and chromatin condensation status) and biochemical parameters (mitochondrial activity, ferric reducing antioxidant power, and malondialdehyde [MDA]) were determined at various stages of cryopreservation. Results: Sperm motility, plasma membrane integrity, viability, acrosome integrity, and chromatin condensation were recorded highest (p < 0.05) with 15 mM quercetin compared with 5, 10, and 20 mM quercetin and control at post-dilution, cooling, equilibration, and freeze-thawing. Nevertheless, mitochondrial activity and antioxidant potential were recorded highest with 15 mM quercetin compared with all experimental extenders at post-equilibration and freeze-thawing. MDA concentration in sperm and seminal plasma were recorded lowest (p < 0.05) in the extender having 15 mM quercetin at post-equilibration and freeze-thawing. Cryopreservation stages showed negative effects (p < 0.05) on semen quality parameters, irrespective of experimental extenders. Conclusions: It is concluded that quercetin (15 mM) supplementation in red fowl extender improves sperm motility, plasma membrane integrity, viability, acrosome integrity, chromatin condensation, and mitochondrial activity by elevating the total antioxidant potential and ameliorating lipid peroxidation during cryopreservation.

Pharmacogenetics of sulfonylurea: Presence of CYP2C9*2, CYP2C9*3 and a novel allele, CYP2C9*61, in Type 2 diabetes patients under sulfonylurea therapy.

CYP2C9 is an important member of the cytochrome P450 gene family involved in the metabolism of 15% of the drugs including an oral antidiabetic agent sulfonylurea. This study aims to investigate the frequency of CYP2C9*2 and CYP2C9*3 alleles of the gene in the sulfonylurea treated diabetic subjects in Pakistan. Briefly, total 105 patients were included in the study and segregated as control (24) and test (81) based on the clinical manifestations after taking sulfonylurea. Genomic DNA was extracted from blood of the subjects and amplified using CYP2C9 specific primers for exon 3 and exon 7 and then subjected to DNA sequencing. Alignment of the sequences with the reference sequence shows presence of CYP2C9*3/*3, CYP2C9*1/*3 and CYP2C9*1/*2 genotypes in the test cases but only the latter two were found in the control cases. In addition a novel allele, CYP2C9*61 in the heterozygous state, was also identified frequently in the test cases. Molecular structure comparison also showed variations in the structural features of protein encoded by the allelic variants. To the best of our knowledge, the present data is the first report for CYP2C9 allelic variations in the indigenous diabetic subjects and also report the existence of novel allelic variant of CYP2C9, CYP2C9*61.

Analysis of Inflammatory Gene Expression Profile of Peripheral Blood Leukocytes in Type 2 Diabetes.

The pathophysiology of type 2 diabetes (T2DM) is associated with perturbation of innate immune response. Several studies indicated alteration of pro-inflammatory and anti-inflammatory cytokines, chemokines and other mediators of innate immune response in T2DM. This study was designed to perform quantitative PCR-based expression profiling of genes involved in inflammation (i.e. CASP1, CASP5, CCL5, CXC11, CCR5, NF-Κb, IL-4, PPARG and PGC1α) in peripheral blood leukocytes of T2DM patients. The T2DM patients are often prescribed with metformin and insulin while metformin has also been reported to possess anti-inflammatory activity. To address the question whether metformin exerts any effect on inflammatory mediators in bloodstream, human subjects in this study were divided into four groups on the basis of medication they were taking during last 6 month. These groups included NT-T2DM (T2DM patients not taking medication, n = 34), Met-T2DM (T2DM patients taking metformin, n = 33), INS-T2DM (T2DM patients taking insulin, n = 15) and NGT (normoglycemic subjects, n = 34) groups. Differential expression of gene transcripts at a cutoff of fourfold was considered significant. In the NT-T2DM group, transcripts of inflammation-related genes (i.e. CASP1, CASP5, CCL5, CCR5 and NF-kB) were up-regulated while transcripts of PPARG and PGC1α genes were down-regulated compared to NGT group. On the other hand, down-regulation of CASP1, CASP5, CCL5, CCR5 and NF-kB transcripts was evident in Met-T2DM and INS-T2DM groups when compared to the NT-T2DM group. The Met-T2DM group and INS-T2DM group showed a significant difference in the transcript level of CASP1 and CCL5 which are more down-regulated in the Met-T2DM group compared to INS-T2DM group. These findings indicated that (a) in T2DM, expression of inflammation-related genes is up-regulated and (b) anti-inflammatory activity of metformin appears to be independent of its anti-hyperglycemic activity.

Immunohistochemical Expression of CD10, BCL6 and MUM1 in Differentiating Diffuse Large B Cell Lymphoma Subtypes.

OBJECTIVE: To determine the immunohistochemical expression of CD10, BCL6 and MUM1 in differentiating diffuse large B cell lymphoma subtypes. STUDY DESIGN: Descriptive, cross-sectional study. PLACE AND DURATION OF STUDY: Department of Histopathology, Armed Forces Institute of Pathology, Rawalpindi, from November 2014 to May 2015. METHODOLOGY: Newly diagnosed cases of DLBCLon H&E stain as well as IHC markers, according to WHO blue book 2008, were included in the study. Patients' gender, age and site of lymphoma were noted. DLBCLsubtypes (GCB and activated type or non-GCB) were assessed based on IHC expression of CD10, BCL6 and MUM1 and the results were recorded. The data were analyzed by using computer software program SPSS version 20. Descriptive statistics, frequencies and percentages were calculated. RESULTS: Out of 96 patients, 79 (82%) were male and 17 (18%) were female. Mean age was 54.66 ±16.73 years. Thirty-six (37.5%) cases showed positivity for CD10 and BCL6 both (GCB type), whereas MUM1 was positive in 60 (62.5%) cases (non-GCB type or activated type). Asignificant statistical association was seen between expression of IHC markers (CD10, BCL6 and MUM1) and DLBCLsubtypes (GCB and non-GCB type, p<0.001). CONCLUSION: In Pakistani population, the frequency of non-GCB type expressing MUM1 is 62.5%, which is quite high as compared to western countries. It needs to be further explored, because it represents high-risk subsets in which alternative strategies for diagnosis and management should be planned.

Hemodialysis patients profile at Dow University of Health Sciences, Karachi. Pakistan.

OBJECTIVES: To determine the frequency of diseases contributing to End Stage Renal Failure (ESRF) and to determine the frequency of seropositivity for hepatitis B and hepatitis C in our patients. METHODS: This is an observational study of two years duration from January 2012 till December 2013, done at Dow university of Health Sciences. Sample size is 189 by convenient method. Data collection is retrospective. Inclusion criteria includes all patients ever hemodialysed at DIMC with age 14 or above. Exclusion criteria is age below 14. Data maintained and analyzed on SPSS version 16. All categorical data in percentages and numeric data is given in frequencies and mean with Standard deviation. RESULT: Total number of patients included in study were 189, Males were 94/189 (49.7%), females were 95/189 (50.3%), Male to female ratio was 0.98: 1.0. Mean age was 51.88+15.2, range was14-86 years. Patients started on Hemodialysis were found to have hypertension in 40.2%, both diabetes and hypertension was present in 42.8%, diabetes alone in 3.1% of patients as likely etiology of renal failure. Seropositivity for HBV was found 4/189(2.1%) and HCV in 31/189(16.4%) at initiation of Hemodialysis. CONCLUSION: Hypertension alone is an important disease found in patients with renal failure as likely cause followed by diabetes. Hepatitis C positivity at start of hemodialysis is 16%.