Detection of quantitative trait loci for mungbean yellow mosaic India virus (MYMIV) resistance in mungbean (Vigna radiata (L.) Wilczek) in India and Pakistan.

Yellow mosaic disease (YMD) is one of the major diseases affecting mungbean (Vigna radiata (L.) Wilczek). In this study, we report the mapping of the quantitative trait locus (QTL) for mungbean yellow mosaic India virus (MYMIV) resistance in mungbean. An F8 recombinant inbred line (RIL) mapping population was generated in Thailand from a cross between NM10-12-1 (MYMIV resistance) and KPS2 (MYMIV susceptible). One hundred and twenty-two RILs and their parents were evaluated for MYMIV resistance in infested fields in India and Pakistan. A genetic linkage map was developed for the RIL population using simple sequence repeat (SSR) markers. Composite interval mapping identified five QTLs for MYMIV resistance: three QTLs for India (qYMIV1, qYMIV2 and qYMIV3) and two QTLs for Pakistan (qYMIV4 and qYMIV5). qYMIV1, qYMIV2, qYMIV3, qYMIV4 and qYMIV5 explained 9.33%, 10.61%, 12.55%, 21.93% and 6.24% of variation in disease responses, respectively. qYMIV1 and qYMIV4 appeared to be the same locus and were common to a major QTL for MYMIV resistance in India identified previously using a different resistant mungbean.

Ambulatory Blood Pressure Monitoring versus Office Blood Pressure Monitoring to Identify the True Hypertension Status of Living Kidney Donors.

Ambulatory blood pressure monitoring (ABPM) is a reliable modality and is preferred over office blood pressure monitoring (OBPM) for detecting hypertension. However, despite its advantages, the utilization of 24-h ABPM in evaluating living kidney donors has not been universally adopted by transplant centers, partly because of the lack of data about the utility of ABPM. This study aimed to identify patients with masked and white-coat hypertension, thereby ensuring appropriate identification of their true hypertension status and assessments of the risk to donors. This study included 73 potential living kidney donors. BP was measured in the office using a standardized protocol as well as by ABPM. Detailed clinical and biochemical parameters were assessed. Target organ damage was assessed in all the donors by assessing proteinuria, hypertensive retinopathy, and echocardiography. Out of the 73 donors, 64.4% were females and 35.6% were males. The average age of individuals in our donor population was 42.0 ± 11.28 years. In total, 31.5% were detected to be hypertensive by OBPM. With ABPM, only 21.9% of donors were hypertensive. The overall prevalence of white-coat hypertension was 30.4%; that of masked hypertension was 6.0%. In donors diagnosed as hypertensive by OBPM, three individuals were identified as having target organ damage. However, two additional donors who were initially missed as hypertensive using OBPM had target organ damage. OBPM overestimated the prevalence of hypertension compared with ABPM. ABPM is the better modality in terms of diagnosing white coats and masked hypertension. ABPM also more reliably correlates with target organ damage than OBPM.

Application of the piecewise rational quadratic interpolant to the AUC calculation in the bioavailability study.

This study presents an application of the piecewise rational quadratic interpolant to the AUC calculation in the bioavailability study. The objective of this work is to find an area under the plasma concentration-time curve (AUC) for multiple doses of salbutamol sulfate sustained release tablets (Ventolin oral tablets SR 8 mg, GSK, Pakistan) in the group of 24 healthy adults by using computational mathematics techniques. Following the administration of 4 doses of Ventolin tablets 12 hourly to 24 healthy human subjects and bioanalysis of obtained plasma samples, plasma drug concentration-time profile was constructed. The approximated AUC was computed by using computational mathematics techniques such as extended rectangular, extended trapezium and extended Simpson's rule and compared with exact value of AUC calculated by using software - Kinetica to find best computational mathematics method that gives AUC values closest to exact. The exact values of AUC for four consecutive doses of Ventolin oral tablets were 150.58, 157.81, 164.41 and 162.78 ngxh/mL while the closest approximated AUC values were 149.24, 157.33, 164.25 and 162.28 ngxh/mL, respectively, as found by extended rectangular rule. The errors in the approximated values of AUC were negligible. It is concluded that all computational tools approximated values of AUC accurately but the extended rectangular rule gives slightly better approximated values of AUC as compared to extended trapezium and extended Simpson's rules.

Protective role of ginseng against gentamicin induced changes in kidney of albino mice.

BACKGROUND: Use of gentamicin is now limited due to its toxic effects, mainly on kidney and vestibular system. Herbal products including ginseng has been reported to possess protective effects against drugs induced nephrotoxicity in experimental animals. The current investigation was designed to evaluate the effects of ginseng on gentamicin induced nephrotoxicity. METHODS: Eighteen male albino mice of 6-8 weeks age, were divided into 3 groups. Group-A served as control and was given normal mouse diet; Group-B was given 80 mg/Kg/day of gentamicin intraperitoneally dissolved in 1 ml of distilled water for fifteen days. Group-C was given 80 mg/Kg/day of gentamicin intraperitoneally dissolved in 1 ml of distilled water along with 100 mg/Kg/day of ginseng orally dissolved in 1 ml of distilled water, also for fifteen days. At the end of the experiment, blood was drawn from each animal by cardiac puncture for renal function tests. Each animal was then sacrificed and kidneys removed for routine histological studies. RESULTS: In group B, weight of the animals and kidneys decreased and there was significant increase in mean serum urea, creatinine and intraluminal diameter (p < 0.001) of proximal convoluted tubules as compared to the controls (group-A). Moderate to severe necrotic and degenerative changes in proximal convoluted tubules were seen in this group. When the Ginseng and gentamicin were given together (group-C), a statistically significant improvement in the mean body and kidney weight along with improvement in renal function tests and tubular diameter were seen (p < 0.001). CONCLUSION: It appears that Ginseng has some protective role against gentamicin induced nephrotoxicity.

A Comparative Study of Epidural Anesthesia with Dexmedetomidine Infusion versus General Anesthesia for Percutaneous Nephrolithotomy.

Background: There has been considerable debate regarding the ideal anaesthetic technique for Percutaneous Nephrolithotomy (PCNL). PCNL is usually performed under general anaesthesia (GA) in prone position. The prone position under GA is associated with various complications. To address these complications, our study was carried out to determine whether epidural anaesthesia [EA] with dexmedetomidine sedation can be a better alternative to GA for PCNL. Aims and Objectives: To compare the efficacy and feasibility of performing PCNL under EA in combination with dexmedetomidine infusion. Methods and Material: Out of 225 patients observed in this study, 115 patients (group A) underwent PCNL under EA with dexmedetomidine infusion and 110 patients (group B) underwent PCNL under GA. Results: Mean time to the first dose of rescue analgesia was significantly increased in Epidural group (328.17 ± 63.74) compared to GA group (72.09 min, p < .0001) and the mean Visual analog scale [VAS] scores were significantly lower in epidural compared to GA group at different time intervals during the first 24 h after surgery (p<0.05). Patients in epidural group had significantly less post-operative nausea, vomiting (6.1 % vs 13.6 %), and significantly less shivering (12.2% vs 33.6%). Mean patient satisfaction score in epidural group was significantly higher (8.75 ± 1.29 vs 8.14 ± 1.39, p=0.001); however, the mean surgeon satisfaction score was comparable among the two groups (8.76 ± 1.39 in Group A and 8.61 ± 1.35 in Group B, p=0.421). Conclusions: Our study shows that EA is an equally effective alternative to GA for PCNL, with more patient satisfaction, less postoperative pain, early ambulation and postoperative recovery, less systemic analgesic requirements and less adverse effects.

Ginkgo biloba induced malformations in mice.

OBJECTIVE: To determine the gross structural malformations to the mice fetuses of the mothers given Ginkgo biloba during pregnancy. STUDY DESIGN: Experimental study. PLACE AND DURATION OF STUDY: The Experimental Research Laboratory, University of Health Sciences, Lahore, from May 2006 to December 2006. METHODOLOGY: The teratogenic effects of Ginkgo biloba extract (78 mg/kg/day and 100 mg/kg/day) dissolved in water were studied on the gross features of mice fetuses. Three groups (A, B and C) of 6 females each were mated with 2 males in two cages with 3:1 ratio of females to males. The first two groups (A and B) served as experimental and the third (C) was used as a control. Pregnancy was confirmed by a vaginal plug and gravid female mice (6) were separated from the males. Group A was treated with human therapeutic dose (78 ppm) while group B was given a high dose (100 ppm). Group C was given water only. Both experimental groups were given the drug orally throughout the gestational period. Result were compared using ANOVA with significance at p < 0.05. RESULTS: Forty-nine fetuses from B and C groups and 50 fetuses from A group were recovered. There was a significant (p < 0.05) decrease in weight and crown-rump length of fetuses in group B as compared to those from group A and C. Further, fetuses from groups A and C did not show any gross abnormalities, whereas those from group B exhibited a high frequency of malformations including round shaped eye and orbits, syndactyly, malformed pinnae, nostrils, lips and jaws. CONCLUSION: The results obtained substantiate the early finding that Ginkgo biloba can be teratogenic when given to pregnant mothers.

Bisphenol-A induced antioxidants imbalance and cytokines alteration leading to immune suppression during larval development of Labeo rohita.

Recently, the oxidative stress and immunotoxicity biomarkers have been extensively used in embryotoxicity using fish embryos as promising models especially after exposure to chemical-like environmental estrogens. Bisphenol-A (BPA) is an estrogenic endocrine disruptor and is ubiquitous in the aquatic environment. Larvae of Labeo rohita were exposed to low concentrations of BPA (10, 100, 1000 µg/l) for 21 days. Innate immune system, antioxidants parameters, and developmental alterations were used as biomarkers. Exposure to BPA caused developmental abnormalities including un-inflated swim bladder, delayed yolk sac absorption, spinal curvature, and edema of pericardium. Lipid peroxidation increased and activity of catalase (p < 0.05), superoxide dismutase (p < 0.05), and glutathione peroxidase (p < 0.01) decreased after exposure to BPA. Level of reduced glutathione also decreased (p < 0.05) in BPA-exposed group. Lower expression of tumor necrosis factor-α (p < 0.05) and interferon-γ (p < 0.001) was observed in BPA-exposed groups while expression of interleukin-10 increased (p < 0.05) in larvae exposed to 10 µg/l BPA. Moreover, exposure of BPA caused a concentration-dependent increase in expression of heat shock protein 70 (p < 0.05). The present study showed that the exposure to BPA in early life stages of Labeo rohita caused oxidative stress and suppress NF-κB signaling pathway leading to immunosuppression. The results presented here demonstrate the cross talk between heat shock protein 70 and cytokines expression.

Age and gender-related differences in mitral cells of olfactory bulb.

OBJECTIVE: To investigate the age and gender-related differences in mitral cells of the human cadaveric olfactory bulbs. STUDY DESIGN: A cross-sectional study. PLACE AND DURATION OF STUDY: The study was conducted in the Department of Anatomy, University of Health Sciences, Lahore, from August 2005 to July 2007. METHODOLOGY: Sixty olfactory bulbs, 30 each from male and female (age 20-76 years) human cadavers divided into six groups of age and gender-wise were collected from the mortuary of the King Edward Medical University, Lahore. Mitral cells were counted and their diameter was calculated from 10 microm thick cresyl violet stained histological sections. Statistical analysis was done using ANOVA for age-related differences and independent t-test for gender-related differences. RESULTS: There was significant reduction in the number of mitral cells and diameter of their nuclei with age. There was significant decrease in the number of mitral cells in males, between groups I and II (p < 0.001); II and III (p < 0.001); and I and III (p < 0.001); statistically significant decrease also occurred in females, between groups IV and V (p < 0.001); V and VI (p < 0.001); and IV and VI (p < 0.001). In most cases, the distance between individual mitral cells was seen to be much greater than in younger group. In group VI, few mitral cells were observed in the cell layer. There was also significant decrease in the diameter of mitral cell nuclei in males, between groups I and III (p < 0.001); and II and III (p < 0.010); in females, between groups IV and VI (p < 0.001); and V and VI (p < 0.001). No gender-related differences were observed. CONCLUSION: The number of mitral cells and diameter of their nuclei decreased with advancing age.

Male offspring of both diabetic parents have higher insulin resistance and serum leptin levels compared to those with one diabetic parent.

OBJECTIVE: We studied the influence of positive family history (FH) of type 2 diabetes mellitus (T2DM) in male offspring using multiple metabolic and endocrine parameters in order to assess whether one or more of these parameters can be used as indicators for T2DM development later in life. DESIGN: Fifty male subjects with one diabetic parent (ODP) and thirty with both diabetic parents (BDP) were compared with fifty, age-matched, offspring of non-diabetic parents (NDP). Body weight, height, BMI and blood pressure were determined in all subjects. Fasting blood samples were analyzed for glucose, HbA1-c, insulin, C-peptide, leptin and lipid profile. A 2h oral glucose tolerance test (2h-OGTT) was also carried out. Insulin resistance (IR) was assessed by HOMA-IR index. RESULTS: Mean serum levels of glucose (fasting and following 2h-OGTT), C-peptide and leptin in male offspring of diabetic parents were higher than in male offspring of NDP. Mean fasting serum insulin and triglycerides were higher in boys of BDP compared to those of ODP and NDP. HOMA-IR was markedly high in ODP and BDP groups when compared with the NDP group. No stastically significant difference was observed in the HbA1-c values between any of the groups studied. CONCLUSIONS: These results indicate that T2DM associated risk factors are more vigorously expressed in male offspring with a history of diabetes in both parents, thus underscoring the importance of genetic determinants in the onset of T2DM. The results of this study may provide useful indicators of potential susceptibility to T2DM at an early stage of life.

Susceptibility of cotton whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) to diverse pesticides in Pakistan.

The cotton whitefly Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a serious pest of several summer crops in hot and dry climates. Its field populations (Asia II-1 biotype) were assessed for their susceptibility to diverse pesticides by using leaf-dip bioassay. There was no or a very low resistance to amitraz, hexythiazox, and pyridaben during 1992-2015. B. tabaci also exhibited no resistance to endosulfan during 1992-1997 and a very low resistance during 1998-2010, which then rose to a low level during 2011-2015. Chlorfenapyr resistance was very low during 1997-2008 and it reached to a high level during 2009-2011 and to a very high level in 2013 and 2015. Among avermectins, abamectin showed a very low resistance up to 2013 but a high resistance in 2015. Emamectin benzoate also demonstrated a very low resistance up to 2010, but a moderate-to-high resistance during 2011-2015. It may be concluded that the diverse chemistries, having novel modes of action and showing no, very low or low levels of resistance, can be substituted in rotation in the wake of resistance development to conventional insecticides.

Metabolic syndrome: an update on diagnostic criteria, pathogenesis, and genetic links.

Metabolic syndrome (MetS), today a major global public health problem, is a cluster of clinical, metabolic, and biochemical abnormalities, such as central adiposity, hypertension, insulin resistance, and dyslipidemias. These MetS-related traits significantly increase the risk of type 2 diabetes mellitus, adverse cardiac events, stroke, and hepatic steatosis. The pathogenesis of MetS is multifactorial, with the interplay of environmental, nutritional, and genetic factors. Chronic low-grade inflammation together with visceral adipose tissue, adipocyte dysfunction, and insulin resistance plays a major role in the progression of the syndrome by impairing lipid and glucose homeostasis in insulin-sensitive tissues, such as the liver, muscle, and adipocytes. Adipose-derived inflammatory cytokines and non-esterified fatty acids establish the link between central obesity IR, inflammation, and atherogenesis. Various studies have reported an association between MetS and related traits with single-nucleotide polymorphisms of different susceptibility genes. Modulation of cytokine levels, pro-oxidants, and disturbed energy homeostasis, in relation to the genetic variations, is described in this review of the recent literature, which also provides updated data regarding the epidemiology, diagnostic criteria, and pathogenesis of MetS.

Determination of Serum Trace Elements (Zn, Cu, and Fe) in Pakistani Patients with Rheumatoid Arthritis.

Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory disease, which mainly involves the joints. RA is prevalent worldwide with increasing prevalence in elderly people. The mechanism of RA pathogenesis is still undefined, and it is interplaying between genetic susceptibility and environmental factors. Although risk factors for RA are not fully established, various studies have focused on the role of trace elements in association with RA. Trace elements act as co-factors for most of the enzymes, and their deficiency is associated with many untoward effects on human health. The homeostatic alterations in the metabolism of trace elements may partly be due to inflammatory response in RA. The objective of the present study was to determine the serum concentrations and correlation of zinc, copper, and iron in RA patients and healthy controls. The study comprised of 61 RA patients and 61 age- and sex-related healthy individuals of Pakistani population. Serum levels of Zn, Cu, and Fe were measured in all the participants by atomic absorption spectrophotometer. Serum Zn and Fe were significantly reduced in the RA patients than those in the healthy controls. Serum Cu concentrations were found elevated in the RA patients. Correlation studies of trace elements determine that there was negative correlation between Zn and Cu in the RA patients and no correlation in the control group. It is very important to explore the deficiency of essential trace metals in biological samples of the RA patients in different populations which may be helpful for diagnosis and supplementary management of rheumatoid arthritis patients.

Effect of linogliride on hormone release from perfused rat pancreas. Fuel dependence and desensitization by tolbutamide.

We examined the effect of the hypoglycemic drug linogliride on hormone release from the in vitro perfused rat pancreas. Linogliride stimulated insulin release in the absence of glucose either in the presence or absence of a physiological mixture of amino acids. In addition, linogliride inhibited amino acid-induced glucagon release. Half-maximal effects of linogliride on insulin and glucagon release were achieved at concentrations as low as 26 and 3 microM, respectively. The effects of linogliride on hormone release largely resembled those of tolbutamide. In the absence of amino acids, the stimulation of insulin release by linogliride or tolbutamide was transient. When the pancreas had been preperfused for 20 min with tolbutamide, linogliride no longer had an effect on hormone release. Likewise, tolbutamide remained without effect in pancreases preperfused with linogliride. These data suggest that linogliride and tolbutamide may have a similar mechanism of action.

Carbon nanotube-reinforced hydroxyapatite composite and their interaction with human osteoblast in vitro.

As a bone mineral component, hydroxyapatite (HA) has been an attractive bioceramic for the reconstruction of hard tissues. However, its poor mechanical properties, including low fracture toughness and tensile strength, have been a substantial challenge to the application of HA for the replacement of load-bearing and/or large bone defects. In this study, HA is reinforced with high-purity and well-functionalized multiwalled carbon nanotubes (MWCNTs; >99 wt%) having an average diameter of 15 nm and length from 10 to 20 µm. The cellular response of these functionalized CNTs and its composites were examined in human osteoblast sarcoma cell lines. Calcium nitrate tetrahydrate (Ca(NO3)2·4H2O) and diammonium hydrogen phosphate ((NH4)2HPO4) were used to synthesize HA in situ. MWCNTs were functionalized by heating at 100°C in 3:1 ratio of sulfuric acid and nitric acid for 60 min with stirring and dispersed in sodium dodecyl benzene sulfonate by sonication. HA particles were produced in MWCNTs solution by adding Ca(NO3)2·4H2O and (NH4)2HPO4 under vigorously stirring conditions. The composite was dried and washed in distilled water followed by heat treatment at 250°C to obtain CNT-HA powder. Physiochemical characterization of the composite material was carried out using Fourier transform infrared spectroscopy, field-emission scanning electron microscopy, energy-dispersive X-ray spectrometer, and X-ray diffractometer. Furthermore, this study investigates the cytotoxic effects of functionalized-MWCNTs (f-MWCNTs) and its composites with HA in human osteoblast sarcoma cell lines. Human osteoblast cells were exposed with different concentrations of f-MWCNTs and its composite with HA. The interactions of f-MWCNT and MWCNT-HA composites were analyzed by 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. The results indicate no detrimental effect on survival or mitochondrial activity of the osteoblast cells. Cell viability decreased with an increase in CNT concentration indicating that MWCNTs and its composite can be cytotoxic at higher dosages. This result provides further evidence that the bionano interface can be developed for CNT-reinforced HA composites for load-bearing bone implants, drug delivery, and tissue engineering.

A prospective, randomized, double blind study to compare the effects of equiosmolar solutions of 3% hypertonic saline and 20% mannitol on reduction of brain-bulk during elective craniotomy for supratentorial brain tumor resection.

AIMS: The aim of the study was to compare the effect of mannitol (M) and hypertonic saline (HTS) on brain relaxation and electrolyte balance. SETTINGS AND DESIGN: Prospective, randomized, double-blind study. SUBJECTS AND METHODS: A total of 114 patients with American Society of Anesthesiologists status II and III, scheduled to undergo craniotomy for supratentorial brain tumor resection were enrolled. Patients received 5 ml/kg 20% mannitol (n = 56) or 3% HTS (n = 58) at the start of scalp incision. Hemodynamics, fluid balance and electrolytes, were measured at 0, 15, 30, and 60 min and 6 h after infusion. Intensive Care Unit (ICU) stay between the two groups was also recorded. The surgeon assessed brain relaxation on a four-point scale (1 = Relaxed, 2 = Satisfactory, 3 = Firm, 4 = Bulging). Appropriate statistical tests were used for comparison; P < 0.05 was considered significant. RESULTS: Brain relaxation conditions in the HTS group (relaxed/satisfactory/firm/bulging, n = 28/20/5/3) were better than those observed in the M group (relaxed/satisfactory/firm/bulging, n = 17/21/11/9). The levels of serum sodium were higher in the HTS group (P < 0.001). The average urine output was higher in the M group (5.50 ± 0.75 L) than in the HTS group (4.38 ± 0.72 L) (P < 0.005). There was no significant difference in fluid input, ICU stay, and hospital days between the two groups. CONCLUSION: We concluded that HTS provided better brain relaxation than mannitol during elective supratentorial brain tumor surgery, without affecting ICU and hospital stay.

Effect of shilajit on the heart of Daphnia: A preliminary study.

Shilajit is a mineral-rich complex organic compound used in the traditional system of Ayurvedic medicine for treating hypertension and improving the cardiac function with many herbomineral preparations. However, very little experimental evidence is available about its effect on the cardiac function. We used Daphnia as a model organism for observing the effect of shilajit on its heart due to its myogenic properties and its response to number of cardioactive drugs that are known to affect human heart function. Genome of Daphnia shows the strongest homology with the human genome. These characteristics of Daphnia make it an ideal organism for biomedical research. Our results suggest that this complex organic compound lowers the heart beats as its concentration increases from 1.0 to 100 ppm. The beats come to near normal condition at 1000 ppm. Above 1000 ppm, the beats are very fast and impossible to count. These results indicate a negative chronotropic effect on the Daphnia heart at low concentrations and a positive chronotropic effect to arrhythmia and finally failure at increasing higher concentrations of shilajit.

Frequency distribution of PRNP polymorphisms in the Pakistani population.

Prion diseases are neurodegenerative conditions caused by misfolding of a normal host-encoded prion protein (PrPC) into pathogenic scrapie prion protein (PrPSc). In human prion diseases, the M129V prion protein polymorphism is known to confer susceptibility to the disease, determines PrPSc conformation and alters clinicopathological phenotypes. To date, all clinicopathologically confirmed cases of a variant form of Cruetzfeldt-Jacob disease (vCJD) have been 129MM homozygotes. There is also predominance of 129MM homozygotes in sporadic CJD (sCJD). No information regarding prion disorders is available from Pakistan. Although only invasive procedures like brain biopsy can confirm the diagnosis of prion disorders, testing a corresponding human population for variation in the prion protein gene (PRNP) may provide some insights into the presence of these disorders in a locality. The current study therefore aimed at exploring the genetic susceptibility of Pakistani population to CJD. A total of 909 unrelated individuals including 221 hemophiliacs representing all 4 major provinces of Pakistan were screened for M129V polymorphism and insertions or deletions of octapeptide repeats (OPRIs/OPRDs) using Polymerase Chain Reaction coupled with Restriction Fragment Length Polymorphism (PCR-RFLP). Concordance of the results of some PCR-RFLP reactions was also confirmed by dideoxy automated Sanger sequencing. The frequencies of M129V alleles (129M and 129V) and genotypes (129MM, 129MV and 129VV) were found in all 909 individuals to be 0.7101, 0.2899, 0.5270, 0.3663 and 0.1067, respectively. Deletion of 1 octapeptide repeat (1-OPRD) was detected in heterozygous state in PRNP of 10 individuals and in homozygous state in 1 individual. An insertion of 3 octapeptide repeats (3-OPRI) was found in 1 individual and an insertion of 1 octapeptide repeat (1-OPRI) in two individuals. Both 3-OPRI and 1-OPRI were present in heterozygous state and were linked to 129M allele. There were no significant χ2 differences between M129V allelic and genotypic frequencies of healthy individuals and hemophiliacs. However, M129V allelic and genotypic frequencies differed significantly between Pakistani population and East Asian and Western populations. Non-significant χ2 differences between M129V frequencies of healthy individuals and hemophiliacs suggest that individuals manifesting single gene disorders may provide naturally randomized samples for studies aiming at surveying the genetic variation. The combined excess of 129MM and 129VV homozygosity and the presence of 3-OPRI in 1 individual imply that Pakistani population is susceptible to prion disorders. Cases of prion disorders may exist in Pakistan, albeit at lower annual prevalence than other countries where life expectancy is greater than 65 years.

PRNP gene variation in Pakistani cattle and buffaloes.

Bovine spongiform encephalopathy (BSE) is a neurodegenerative prion protein misfolding disorder of cattle. BSE is of two types, classical BSE and atypical BSE which in turn is of two types, H-type BSE and L-type BSE. Both H-type BSE and L-type BSE are primarily sporadic prion disorders. However, one case of H-type BSE has recently been associated with E211K polymorphism in the prion protein gene (PRNP). Two polymorphisms in the bovine PRNP are also associated with susceptibility to classical BSE: a 23 bp insertion/deletion (indel) in the PRNP promoter region and a 12 bp indel in the first intron. No information regarding BSE susceptibility in Pakistani cattle is available. The present study aimed at achieving this information. A total of 236 cattle from 7 breeds and 281 buffaloes from 5 breeds were screened for E211K polymorphism and 23 bp and 12 bp indels employing triplex PCR. The E211K polymorphism was not detected in any of the animals studied. The 23 bp insertion allele was underrepresented in studied cattle breeds while the 12 bp insertion allele was overrepresented. Both 23 bp and 12 bp insertion alleles were overrepresented in studied buffalo breeds. Almost 90% of alleles were insertion alleles across all studied buffalo breeds. The average frequency of 23 bp and 12 bp insertion alleles across all studied cattle breeds was found to be 0.1822 and 0.9407, respectively. There were significant differences between Pakistani and worldwide cattle in terms of allele, genotype and haplotype frequencies of 23 bp and 12 bp indels. The higher observed frequency of 12 bp insertion allele suggests that Pakistani cattle are relatively more resistant to classical BSE than European cattle. However, the key risk factor for classical BSE is the dietary exposure of cattle to contaminated feedstuffs.

Ethanol induced hepatotoxicity in albino rats.

Twelve male albino rats of 6-8 weeks old, weighing 150-200 gm each were divided into two groups of 6 rats each. Group A was used as control while Group B was given ethanol at a dose of 0.6 ml (0.5 gm)/100 gm/day for 8 weeks. Serum enzymes and liver histology was determined in both groups. Statistically significant increase in the mean enzyme levels, liver weight and volume were observed in the ethanol treated group compared to the controls. Histologically, hepatocytes contained large number of cytoplasmic vacuoles, pyknotic nuclei, and lymphocytic infiltration in treated animals. Ethanol appeared to be hepatotoxic in albino rats.