Ameliorative Influence of Dietary Dates on Doxorubicin-Induced Cardiac Toxicity

Doxorubicin is a commonly used anticancer agent, which may cause cardiac toxicity. The present study designed to evaluate Phoenix dactylofera (dates) in doxorubicin (DXR) induced cardiac toxicity and cardiac remodeling in Wistar albino rats. The experimental rats procured, acclimatized and finally divided into five groups (n = 6). Group I served as normal controls, group II served as disease controls and groups 3, 4 & 5 served as therapeutic groups (Phoenix dactylofera 5%, 10%, and 15% respectively). Cardiac remodeling and toxicity in the rats were induced by administration of DXR (1.25 mg/kg i.p. in 16 divided doses/month). At the end of protocol, effect of Phoenix dactylofera on cardiac remodeling was evaluated by measuring parameters like haemodynamics, heart weight, anatomy, Troponin T, creatine phosphokinase (CPK), creatine phosphokinase-MB (CPK-MB), Lactate dehydrogenase (LDH), serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), calcium ion Ca2+, sodium ion Na+, potassium ion K+, intracellular enzymes like Malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT). The disease control groups showed significantly elevated (p dactylofera significantly (p 2+, Na+, K+ levels to a normal value. Further, the histological studies of the cardiac tissues demonstrated that the normal architecture of the cardiac cells was restored in the animals fed with dietary Phoenix dactylofera as compared to disease controls. The findings show that the administration of Phoenix dactylofera has the potential to prevent the toxicity induced by doxorubicin in the experimental rats.

HCV genotype distribution and possible transmission risks in Lahore, Pakistan

AIM: To investigate the prevalence of hepatitis C virus (HCV) genotypes and their association with possible transmission routes in the general population of Lahore, as the data exclusively related to this city is limited. METHODS: Complete data regarding patient's history, possible route of infection and biochemical tests was collected from the public hospital for 1364 patients. SPSS version 16 windows software was used for data analysis by univariate and multivariate techniques. RESULTS: Age range ≤ 40 years showed high prevalence of HCV infection. HCV genotype 3a was dominant (55.9%), followed by 1a (23.6%), 4a (12.5%), 3b (3.2%), untypable (2.5%), 4b (1.2%) and mixed type (1.2%). Blood transfusion, dental surgery and barber shops were the main risk factors for HCV transmission. Genotype prevalence was independent of age (P = 0.971) and gender (P = 0.122) while risk factors showed a significant association with age (P = 0.000) and genotypes (P = 0.000). We observed an independent association of risk factors and genotype 3a, while patients with genotype 1 and 4 were mostly infected due to dental surgery blood transfusion and barber shops. Risk factors of intravenous drug use and sexual exposure were exclusively found in ≤ 40 years age group. CONCLUSION: An increase in genotypes 1a and 4a suggest migration of people, possibly from Balochistan and the northern war-zone area. Government should focus on public education regarding infection routes.

In vitro inhibitory analysis of consensus siRNAs against NS3 gene of hepatitis C virus 1a genotype

Objective: To explore inhibitory effects of genome-specific, chemically synthesized siRNAs(small interference RNA) against NS3 gene of hepatitis C virus(HCV) 1a genotype in stable Huh-7(human hepatoma) cells as well as against viral replication in serum-inoculated Huh-7 cells. Methods: Stable Huh-7 cells persistently expressing NS3 gene were produced under antibiotic gentamycin(G418) selection. The cell clones resistant to 1 000 μg antibiotic concentration(G418) were picked as stable cell clones. The NS3 gene expression in stable cell clone was confirmed by RT-PCR and Western blotting. siRNA cell cytotoxicity was determined by MTT cell proliferation assay. Stable cell lines were transfected with sequence specific siRNAs and their inhibitory effects were determined by RT-PCR, real-time PCR and Western blotting. The viral replication inhibition by siRNAs in serum inoculated Huh-7 cells was determined by real-time PCR. Results: RT-PCR and Western blot analysis confirmed NS3 gene and protein expression in stable cell lines on day 10, 20 and 30 post transfection. MTT cell proliferation assay revealed that at most concentrated dose tested(50 nmol/L), siRNA had no cytotoxic effects on Huh-7 cells and cell proliferation remained unaffected. As demonstrated by the siRNA time-dependent inhibitory analysis, siRNA NS3-is44 showed maximum inhibition of NS3 gene in stable Huh-7 cell clones at 24(80%, P=0.013) and 48 h(75%, P=0.002) post transfection. The impact of siRNAs on virus replication in serum inoculated Huh-7 cells also demonstrated significant decrease in viral copy number, where siRNA NS3-is44 exhibited 70%(P<0.05) viral RNA reduction as compared to NS3-is33, which showed a 64%(P<0.05) decrease in viral copy number. siRNA synergism(NS3-is33 + NS3-is44) decreased viral load by 84%(P<0.05) as compared to individual inhibition by each siRNA(i.e., 64%–70%(P<0.05) in serum-inoculated cells. Synthetic siRNAs mixture(NS5Bis88 + NS3-is33) targeting different region of HCV genome(NS5B and NS3) also decreased HCV viral load by 85%(P< 0.05) as compared to siRNA inhibitory effects alone(70% and 64% respectively, P<0.05). Conclusions: siRNAs directed against NS3 gene significantly decreased m RNA and protein expression in stable cell clones. Viral replication was also vividly decreased in serum infected Huh-7 cells. Stable Huh-7 cells expressing NS3 gene is helpful to develop anti-hepatitis C drug screening assays. siRNA therapeutic potential along with other anti-HCV agents can be considered against hepatitis C.

Source Sector Contributions to Aerosol Levels in Pakistan

Urban air pollution in Pakistan is a serious challenge and it causes significant damage to human health and ecosystems. This paper presents a modelling study using the Weather Research and Forecasting Model coupled with Chemistry(WRF-Chem) to simulate the spatial distributions and temporal variations of aerosol concentrations over Pakistan, focusing on contributions of domestic emission sectors(transport, industry, residential, and energy) to mass concentrations of sulfate(SO2–4), nitrate(NO–3), ammonium(NH+4), black carbon(BC), and organic carbon(OC) during the months of January, April, July, and October in 2010. Sensitivity studies indicate that, averaged over January, April, July, and October of 2010, energy and industry sectors have the largest contributions to SO2–4 concentrations, each of which contributes about 10%?20% to SO2– 4over the polluted eastern Pakistan. The contributions from residential and transport sectors to NO–3 concentrations reach 40%?50% in central Pakistan. The residential sector has the highest contribution of 50%–80% to BC and OC loading in northeastern and southern Pakistan. Examination of sector contributions to aerosol levels in Lahore, the most polluted city in Pakistan, suggests that reductions in emissions in the residential sector should be an efficient measure for improving particulate matter air quality in this region.

Inhibition of Hepatitis C Virus Genotype 1a Non-Structural Proteins by Small Interference RNA in Human Hepatoma Cell Lines

Hepatitis C virus (HCV) infection and associated liver diseases are still challenging and represent a significant health care burden around the world. Although, the treatment strategies have been improved by the development of novel direct-acting antivirals, but such therapeutic options are still expensive and beyond the financial range of the most infected individuals in developing or even in resource replete countries. It demands an urgent need to search novel and improved alternate treatment strategies to treat the infection. The present study was aimed to develop an in vitro stable cell culture system, persistently expressing HCV genotype 1a non-structural genes and to characterize the inhibitory effects of synthetic siRNAs (short interference RNA) directed against the most conserved regions of nonstructural genes in an in vitro cell culture model. The continuous expression of nonstructural genes for more than 30 days post transfection was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis in stable human hepatoma cell line (Huh-7). The gene expression studies revealed significantly reduced gene expression of HCV nonstructural genes (i.e., NS2, NS4A and NS5A) both at mRNA and protein levels when treated against genome specific synthetic siRNAs in stable cell lines (51%, 47% and 54% respectively, p < 0.05). Similarly, a vivid decrease in HCV viral titer was exhibited by synthetic siRNAs in an in vitro viral replicate cell culture model (58%, 48% and 50%, respectively, p < 0.05) determined by quantitative Real-Time PCR (qPCR). Our data indicate that siRNA mediated gene silencing may be considered a promising alternate treatment strategy against HCV in combination with other effective therapeutic regimens in future.

Determination of the Potency of Extracted, Purified and Formulated Insulin from the Pancreatic Organs of the Sudanese Beef Cattle

The treatment of type 1 diabetes is mainly dependent on insulin therapy and current formulated insulin formulations are used for its control all over the world. The presented study was designed to evaluate the potency of extracted, purified and formulated insulin from the pancreatic organs of the Sudanese beef cattle. Twenty healthy rabbits were used to conduct the study following subcutaneous administration of the sample insulin, to determine the hypoglycemic effect and to analyze the potency of the testing insulin by the hypoglycemic seizure method, blood sugar method and glucose enzymatic colorimetric test (GOD-PAP) respectively. The potency of the injected insulin samples was estimated by comparing the variation in blood glucose levels produced in the treated animals with that produced by a standard insulin preparation under the suitable conditions of the blood sugar method. The results revealed that the potency of the testing beef insulin samples was slightly higher (i.e., 2.2 USP units/ml, 9%) compared to the standard and assumed potency of the prepared insulin preparations (i.e., 1-2 USP units/ml) which indicated that the solvents and diluents used to prepare the assay dilution might be of higher potency and must be diluted to such an extent that the testing insulin potency must be compatible with the standard dilutions. Furthermore, to determine the choice of an assay to analyze the potency of insulin preparations, not only the accuracy of the result but also the purpose for which the test is to be used and the time limit must be taken into consideration.

Determination of the Stability Studies of the Sudanese Camel Insulin

The main objective of the presented study was to characterize the stability of the Sudanese camel insulin after 6 months of its extraction, purification and formulation from the fresh pancreatic glands of the camel, slaughtered for local and export consumption. The stability and purity of the formulated insulin samples were compared to standard insulin sam-ples that of the leading manufacturing companies using some analytical techniques such as HPLC, gel electrophoresis and atomic absorption. In another part of the study, the direct transfer method was used to accomplish sterility test by complete immersion of the insulin samples into thioglycollate and soybean medium. The data were presented as mean ± S.E.M (standard error of means) for the comparison of zinc (mg/units) and nitrogen (in percentage) concentrations in standard and testing camel insulin samples, respectively. Similarly, the linear equation was derived and the coefficient factors for standard and testing insulin samples were compared to determine the peak area and the concentrations of the camel insulin samples (mean ± S.E.M) after HPLC elution. The P value,

Zinc Chloride Protects against Streptozotocin-Induced Diabetic Nephropathy in Rats

Diabetes mellitus (DM) is a health problem affecting millions of individuals worldwide. Diabetic nephropathy (DN), as a significant complication of DM, has become the most common cause of endstage renal failure. Oxidative stress constitutes the key and common events in the pathogenesis of DN and antioxidants may play a beneficial role in its prevention. This study was conducted to investigate the effect of Zinc Chloride on streptozotocin-induced diabetic nephropathy in rats compared to Gliclazide, a reference antidiabetic agent. Results showed that Zinc Chloride was able to control STZ-induced DN in rats as it normalized the elevated blood pressure, the increased insulin release, and the decreased blood glucose level. Zinc Chloride also improved kidney function as determined by the restoration of blood urea and creatinine level. Finally, Zinc Chloride was able to boost the antioxidant defenses of the kidney by increasing the reduced glutathione content and decreasing lipid peroxides content in addition to significantly decreasing kidney nitric oxide content compared to diabetic control rats. These results suggest that exposure to Zinc Chloride can protect from diabetic nephropathy and can be used as an adjuvant approach to treatment and prevention of renal damage.