Validation of genome-wide association studies (GWAS)-identified type 2 diabetes mellitus risk variants in Pakistani Pashtun population / Journal of the ASEAN Federation of Endocrine Societies

Objective@#Recent GWAS largely conducted in European populations have successfully identified multiple genetic risk variants associated with Type 2 Diabetes Mellitus (T2DM). However, the effects conferred by these variants in the Pakistani population have not yet been fully elucidated. The objective of this study was to examine European GWAS- identified T2DM risk variants in the Pakistani Pashtun population to better understand the shared genetic basis of T2DM in the European and Pakistani cohorts.@*Methodology@#A total of 100 T2DM patients and 100 healthy volunteers of Pashtun ethnicity were enrolled in this study. Both groups were genotyped for 8 selected single nucleotide polymorphisms (SNPs) using the Sequenom MassARRAY® platform. The association between selected SNPs and T2DM was determined by using appropriate statistical tests.@*Results@#Of the 8 studied SNPs, 5 SNPs, SLC30A8/ rs13266634 (p=0.031, OR=2.13), IGF2BP2/ rs4402960 (p=0.001, OR=3.01), KCNJ11/ rs5219 (p=0.042, OR=1.78), PPARG/ rs1801282 (p=0.042, OR=2.81) and TCF7L2/ rs7903146 (p=0.00006, 3.41) had a significant association with T2DM. SNP GLIS3/ rs7041847 (p=0.051, OR=2.01) showed no sufficient evidence of association. SNPs KCNQ1/ rs2237892 (p=0.140, OR=1.61) and HHEX/IDE/ s1111875 (p=0.112, OR=1.31) showed opposite allelic effects and were not validated for T2DM risk in the study population. Among the studied SNPs, TCF7L2/ rs7903146 showed the most significant association.@*Conclusion@#Our study finding indicates that selected genome-wide significant T2DM risk variants previously identified in European descent also increase the risk of developing T2DM in the Pakistani Pashtun population.

Comparative pharmacokinetics of omeprazole and its metabolites in poor and extensive metabolizer Pakistani healthy volunteers and a review of different studies

This study was designed to evaluate a comparative single dose [40mg] pharmacokinetics [PK] of Omeprazole [OMP] and its two metabolites, 5-hydroxy Omeprazole [5-OH-OMP] and Omeprazole sulphone [OMP-S] in poor [PM] and extensive [EM] metabolizer Pakistani healthy adult volunteers. The frequency of CYP2C19 and CYP3A4 varies widely in different populations. The present study was conducted to evaluate the PK of OMP and its two metabolites in Pakistani population and to review different studies conducted after administration of single dose of OMP. Twenty two subjects were enrolled in this study and divided into two groups. The CYP2C19 phenotyping was evaluated by the metabolic ratio of OMP to 5-OH-OMP. It was a single dose, open label study and the blood samples from subjects were collected at different time intervals until 24 hours. The PK parameters were calculated using the PK-summit software. The metabolic ratio of area under the plasma concentration-time curve AUCOMP/5-OH-OMP was 1.86 +/- 0.572 and13.84 +/- 2.504 for EM and PM, respectively; maximum plasma concentration [Cmax] of OMP was increased by two folds for PM while the AUC? was increased by 3 folds; the Cmax and AUC? of 5-OH-OMP decreased for PM by 2 folds while there was 3 fold increase observed in the Cmax and AUC? of OMP-S. The PK of OMP and its metabolites in different populations were also discussed, and issues regarding CYP2C19 and CYP3A4 genotyping were also extensively reviewed. In EM of CYP2C19 the concentration of 5-OH-OMP is higher while that of OMP-S is lower. This study as well as reported studies reveals that in PM of CYP2C19 more drugs are available for CYP3A4 to be metabolized. A correlation between CYP2C19 EM and PM activity with CYP3A4 needs to be established

Antibacterial and antifungal activities of leaf extract of Achyranthes aspera [Amaranthaceae] from Pakistan

Alcoholic extract and various fractions of Achyranthes aspera leaves, traditionally used in Pakistan for treatment of infectious diseases was screened for in vitro antibacterial and antifungal activity. The chloroform and butanol fractions were found to be the most active among the fractions, showing considerable antibacterial activity against Shigella flexneri and Escherichia coli. The highest activity was found in the ethylacetate fraction [17 mm zone of inhibition] against gram-negative [Salmonella typhi] bacteria, with MIC value as 0.29 mg/mL. In antifungal screening, moderate activity was shown by the chloroform fraction [50 % inhibition] against Microsporum cams, with MIC value as 0.25mg/mL. Considerable level of antifungal activity was depicted by crude extract, hexane and butanol fractions against Aspergillus flavus and Microsporum canis. The ability of various extracts of Achyranthes aspera to inhibit different strains of fungi and bacteria indicates its potential use for the treatment of microbial infections

Evaluation of anti-inflammatory activity of selected medicinal plants of Khyber Pakhtunkhwa, Pakistan

In present study, the anti-inflammatory potential of three medicinal plants, Xanthium strumarium, Achyranthes aspera and Duchesnea indica were evaluated, using both in vitro and in vivo assays. Carrageenan induced hind paw edema model was used to carry out the in vivo anti-inflammatory activity, while for in vitro screening lipoxygenase inhibition assay was used. Crude extract of all the selected plants depicted significant [p

Screening of selected medicinal plants for their enzyme inhibitory potential - a validation of their ethnopharmacological uses

In present study four medicinal plants namely Valeriana wallichii, Xanthium strumarium, Achyranthes aspera and Duchesnea indica belonging to different families were collected in Khyber Pakhtunkhwa province and crude extract and subsequent fractions were analyzed for their inhibitory potential against acetylcholinesterase, butyrylcholinesterase and alpha-glucosidase enzymes. Valeriana wallichii, Xanthium strumarium and Achyranthes aspera were significantly active against cholinesterases. Chloroform and ethylacetate fractions of Valeriana wallichii exhibited significant activity against acetylcholinesterase [IC50: 61microg/ml] and butyrylcholinesterase enzymes [IC50: 58microg/ml], respectively. Similarly ethylacetate fraction of Achyranthes aspera showed significant activity against acetylcholinesterase [IC50: 61 microg/ml] and butyrylcholinesterase enzymes [IC50: 61 microg/ml], respectively. In case of alpha-glucosidase enzyme, the chloroform fraction of Xanthium strumarium exhibited significant inhibitory activity [IC50: 72 microg/ml] as compared to the standard compound acarbose [IC50: 483 microg/ml]. Duchesnea indica showed no such activities.

Anti-inflammatory activity of the topical preparation of Valeriana wallichii and Achyranthes aspera leaves

In vivo and in vitro screening of anti inflammatory activity of Valeriana wallichii and Achyranthes aspera leaves crude extract was performed, using standardized procedures. Methanolic crude extract topical formulation [cream] of Valeriana wallichii and Achyranthes aspera leaves [Family Valerianaceae and Amaranthaceae respectively], were screened for their anti-inflammatory activity, through "Carrageenan induced hind paw edema" test, for their effect on the acute and chronic phase inflammation models in male Wistar rats. Methanolic extract and its fractions were also evaluated for their in vitro anti-inflammatory activity using lipoxygenase inhibition assay. Leaves of Valeriana wallichii showed significant [p<0.001], dose dependant anti inflammatory activity, comparable with that of the standard, in animal model. The ethyl acetate fraction of Valeriana wallichii also showed considerable [IC 50=73 +/- 0.36] in vitro antiinflammatory activity as compared to standard [6.11 +/- 0.02]. Similarly Achyranthes aspera leaves showed relatively weak [p>0.05] in vivo anti- inflammatory activity. However, its activity was comparable with that of standard at 10% concentration after 5 hrs of carrageenan injection. This activity was present in ethyl acetate fraction during in vitro screening [IC 50=76 +/- 0.14] as compared to that of standard [IC 50=6.11 +/- 0.02]. The combined in vitro and in vivo Antiinflammatory screening shows that the ethyl acetate fraction of the crude extract of Valeriana wallichii and Achyranthes aspera can be used for the isolation of new Anti-inflammatory lead compounds

Metal analysis, phytotoxic, insecticidal and cytotoxic activities of selected medicinal plants of Khyber Pakhtunkhwa

In the present study four medicinal plants traditionally used in Pakistan for treatment of various ailments were evaluated for their heavy metals content, insecticidal, cytotoxic and phytotoxic actions. The metals like Cr, Cu, Zn, Mn, Ni, Pb, Fe and Co were determined in crude extract and various fractions. Soil samples were also tested for heavy metals to determine assimilation of any metal by the plant. Lead, chromium, copper, nickel and cobalt exceeded the permissible limit in most of the tested samples while the concentration of zinc, manganese and iron was within the permissible limit. Chloroform fraction from Achyranthes aspera and ethyl acetate fraction from Duchesnea indica showed significant phytotoxic activities. Crude extract and chloroform fraction from Xanthium strumarium showed insecticidal activity comparable to that of permethrin and thus could be a significant source of natural insecticide. The butanol fraction from X. strumarium showed significant cytotoxicity with LC[50] 1.9306 microg/ml, having mortality rate 93% at highest dose, while the crude extract from Valeriana wallichii showed 90% mortality rate [LC[50] 4.9730 microg/ml] at highest dose. However, the extracts from other plants were not effective against the brine shrimps tested

Antimicrobial and anti-inflammatory activities of leaf extract of Valeriana wallichii DC

Valeriana wallichii DC [Valerianaceae] is one of the most widely used traditional remedies for various complications associated with nervous system and digestion. No antimicrobial and anti-inflammatory studies have so far been carried out on the aerial parts of the plant. The present work was focused to evaluate the antimicrobial [antifungal and antibacterial] and anti-inflammatory properties of Valeriana wallichii using reported methods. Chloroform fraction [VW-2] and hexane fraction [VW-3] exhibited significant activity against Staphylococcus aureus and Bacillus subtilus, respectively. The chloroform fraction [VW-2] showed significant activity against Staphylococcus aureus with 0.27 mg/ml MIC, where 0.31 mg/ml MIC was deduced for hexaune fraction [VW-3] against Bacillus subtilus. Hexaune fraction [VW-3] was also found to be the most potent inhibitor of Microsporum canis, showing 70% inhibition with an MIC value of 0.19 mg/ml. Considerable inhibitory activity was also observed for Chloroform fraction [VW-2] and water fraction [VW-6] against Microsporum canis and Aspergillus flavus. A remarkable anti-inflammatory like activity was observed for the crude extract at a dose of 200 mg/kg at all observed durations. Other doses of the sample also showed excellent activity. Looking to these results it may be concluded that Valeriana wallichii may be a potential source for activity guided isolation of natural products with antimicrobial and anti-inflammatory-like properties