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.

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.

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.

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.

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.

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.

Ginkgo biloba effects on mice fetal liver / Efectos de la ginkgo biloba en el hígado fetal de ratón

Ginkgo biloba is considered to be an alternative drug for various indications; unfortunately very few studies are available on its side effects. This present study describes the harmful effects of Ginkgo biloba on developing fetal liver. Two experimental groups of six pregnant female mice each were given Ginkgo biloba at human therapeutic dose (A) and a higher dose (B) throughout the gestation period. A third group (C) was taken as a control and given distilled water only. Fetal livers were examined and the effects of the drug observed. There were signs of congestion and fatty change along with dilatation of sinusoids in a dose dependent manner concluding that Ginkgo biloba affects fetal liver.

La Ginkgo biloba es considerada, en varias indicaciones, como un medicamento alternativo; sin embargo, existen pocos reportes disponibles sobre sus efectos secundarios. Este estudio describe los efectos nocivos de Ginkgo biloba en el desarrollo del hígado fetal. Dos grupos experimentales de 6 ratones hembras preñadas recibieron Ginkgo biloba en la dosis terapéutica humana (A) y una dosis más alta (B) por el período de gestación. Un tercer grupo control (C) recibió agua destilada. Los hígados fetales fueron examinados y observados los efectos de la droga. Hubo signos de congestión y degeneración grasa, junto con la dilatación de sinusoides en función de la dosis. Como conclusión la Ginkgo biloba afecta el hígado fetal.

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.

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.