Stereoselective HPLC assay of donepezil enantiomers with UV detection and its application to pharmacokinetics in rats

This investigation describes a new precise, sensitive and accurate stereoselective HPLC method for the simultaneous determination of donepezil enantiomers in tablets and plasma with enough sensitivity to follow its pharmacokinetics in rats up to 12h after single oral dosing. Enantiomeric resolution was achieved on a cellulose tris (3,5-dimethylphenyl carbamate) column known as Chiralcel OD, with UV detection at 268 nm, and the mobile phase consisted of n-hexane, isopropanol and triethylamine (87:12.9:0.1). Using the chromatographic conditions described, donepezil enantiomers were well resolved with mean retention times of 12.8 and 16.3 min, respectively. Linear response (r > 0.994) was observed over the range of 0.05-2 microg/ml of donepezil enantiomers, with detection limit of 20 ng/ml. The mean relative standard deviation (R.S.D.%) of the results of within-day precision and accuracy of the drug were < or =10%. There was no significant difference (p > 0.05) between inter- and intra-day studies for each enantiomers which confirmed the reproducibility of the assay method. The mean extraction efficiency was 92.6-93.2% of the enantiomers. The proposed method was found to be suitable and accurate for the quantitative determination of donepezil enantiomers in tablets. The assay method also shows good specificity to donepezil enantiomers, and it could be successfully applied to its pharmacokinetic studies and to therapeutic drug monitoring.     

Synthesis of Novel Bi-Heterocycles as Valuable Anti-Diabetic Agents: 2-({5-((2-Amino-1,3-Thiazol-4-yl)methyl)-1,3,4-Oxadiazol-2-yl}sulfanyl)-N-(Substituted)acetamides

Russian Journal of Bioorganic Chemistry - The synthesis of a new series of S-substituted acetamides derivatives of 5-[(2-amino-1,3-thiazol-4-yl)methyl]-1,3,4-oxadiazol-2-thiol were synthesized and...     

Overexpression of nicotinamidase 3 (NIC3) gene and the exogenous application of nicotinic acid (NA) enhance drought tolerance and increase biomass in Arabidopsis

Plant Molecular Biology - Overexpressing Nicotinamidase 3 gene, and the exogenous application of its metabolite nicotinic acid (NA), enhance drought stress tolerance and increase biomass in...     

SLC4A11 mutations causative of congenital hereditary endothelial dystrophy (CHED) progressing to Harboyan syndrome in consanguineous Pakistani families

Background

Autosomal recessive corneal hereditary endothelial dystrophy (CHED) is a rare congenital disorder of cornea. Mutations in SLC4A11 gene are associated with CHED phenotype. CHED is also an early feature of Harboyan syndrome. The aim of the present study was to identify genetic mutations in the SLC4A11 gene in CHED cases belonging to inbred Pakistani families. Furthermore, all homozygous mutation carriers were investigated for hearing deficit.

Methods and results

This study included consanguineous CHED families presented at Al-Shifa Trust Eye Hospital, Rawalpindi, Pakistan from June 2018 to September 2018. DNA was extracted from blood samples. Direct sequencing of SLC4A11 gene was performed. All identified variants were evaluated by in silico programs i.e., SIFT, PolyPhen‐2, and MutationTaster. Pathogenicity of the two identified splice site variants was analyzed by Human Splicing Finder and MaxEnt Scan. Screening of five CHED families revealed a total of three previously un reported (p.Arg128Gly, c.2241-2A?>?T and c.1898-2A?>?C in family CHED19, CHED22 and CHED26 respectively) and two already reported homozygous disease causing variants (p.Arg869Cys and p.Val824Met in family CHED24 and CHED25 respectively) as predicted by mutation taster. All of these variants segregated with disease phenotype and were not detected in controls.

Conclusion

Affected individuals of the five CHED families screened in this study had the disease due to SLC4A11 mutations and progressing to Harboyan syndrome. Identification of previously unreported mutations aid to heterogeneity of SLC4A11 and CHED pathogenesis as well as helped to provide genetic counseling to affected families.

     

Antioxidant effects of Emblica officinalis and Zingiber officinalis on arsenic and lead induced toxicity on Albino rats

It is of interest to document the effect of Emblica officinalis (E. officinalis) and Zingiber officinalae (Z. officinalae) leaf extract on reactive oxygen species, antioxidant potential changes in arsenic and lead-induced toxicity in male rats. We used 8 groups of adult male Wistar rats with 1 control group for this study. The animals were divided into Group I: Control and Group II: Lead and sodium arsenite induced rats (animals were induced for metal toxicity by the combined administration of arsenic (13.8 mg/ kg body weight) and lead (116.4 mg/kg body weight). These doses were administered by gastric intubation during 14 consecutive days using known standard procedures. Arsenic and lead induced rats treated with ethanolic extract of Emblica officinalis (60 mg/kg body weight/day, orally for 45 days) are group III rats. Group IV animals are arsenic and lead induced rats treated orally with ethanolic extracts of E. officinalis (120 mg/kg body weight/day for 45 days). Group V animals are arsenic and lead induced rats treated orally with ethanolic extracts of Z. officinalae (60 mg/kg body weight/day for 45 days). Group VI animals are arsenic and lead induced rats orally treated with ethanolic extracts of Zingiber officinalis (120 mg/kg body weight/day for 45 days). Group VII animals are arsenic and lead induced rats treated orally with ethanolic extracts of E. officinalis and Z. officinalae (60 + 60 mg/kg body weight/day for 45 days). Group VIII animals are arsenic and lead induced rats treated orally with ethanolic extracts of E. officinalis and Z. officinalae (120 + 120 mg/kg body weight/day, orally for 45 days). Normal Control animals were treated orally with ethanolic extracts of E. officinalis (120mg/kg body weight) + Z. officinalae (120mg/kg body weight) for 45 days. The control and experimental animals were then subjected to analysis for oxidative stress markers such as H2O2, *OH, and lipid peroxidation (LPO), antioxidant enzymes in addition to liver and kidney function markers. Results: Arsenic and lead induced rats showed a significant increase in the levels of reactive oxygen species (H2O2, OH* and LPO) with concomitant alterations in the renal and liver tissues. However, enzymic and non-enzymic antioxidant levels were decreased. Nevertheless, an oral effective dose of E. officinalis and Z. officinalae (120 + 120 mg/kg body weight/day increased the antioxidant enzymes and retrieved the altered levels of ROS and LPO that were induced by arsenic and lead. Thus, we show that E. officinalis and Z. officinalae leaf extract exhibits nephroprotective and hepatoprotective role through the restoration of reactive oxygen species and antioxidant enzymes in the kidney and liver tissue of Arsenic and Lead-induced nephrotoxicity and hepatotoxicity in rats. Hence, E. officinalis and Z. officinalae leaf extract are potential therapeutic options for the treatment of metal toxicity-induced kidney and liver diseases.     

Synergistic effects of zinc oxide nanoparticles and various antibiotics combination against Pseudomonas aeruginosa clinically isolated bacterial strains

P. aeruginosa causes mostly both community-acquired and nosocomial infections, which leads to serious therapeutic challenges for treatment and requirement of appropriate therapeutic agent is needed which can combat antibiotic resistance. The research work was performed to investigate the effect of Zinc Oxide nanoparticles (ZnO NPs) in combination with Meropenem, Ciprofloxacin, and Colistin against clinical isolated strains of P. aeruginosa and ATCC 27853 strain.The minimum inhibitory concentration (MIC) of ZnO NPs and the antibiotics (Meropenem, Ciprofloxacin, and Colistin), was determined by the microdilution method and the results of MIC values were ranging between 1 and 16 µg/mL was found to be shown for antibiotics and ZnO NPs found to showed highest MIC values ranging from 2000 to 4000 µg/mL. The fractional inhibitory concentration index (FICI) was calculated using checkerboard method to test the combinations of ZnO NPs and the antibiotics (Meropenem, Ciprofloxacin, and Colistin), and among all the six P. aeruginosa clinical isolated strains P. aeruginosa (MRO-16-3 and MRO-16-4), showed FICI as 0.24 and 0.39 9, whereas P. aeruginosa ATCC 27853 strain showed FICI as 0.41 which indicates synergistic effect with Colistin.The time kill growth curve showed synergistic effect for the combination of Colistin and ZnO NPs against P. aeruginosa (MRO-16-3 and MRO-16-) strains. P. aeruginosa (MRO-16-3) was found to be highly sensitive to Colistin with an MIC of 2 µg/mL, which has shown to reduced bacterial growth to zero colonies after 24 h of incubation.In conclusion, combination of Colistin and ZnO NPs at appropriate dosage intervals might be beneficial as using therapeutic agent in treatment of P. aeruginosa ailments.     

Autosomal Dominant Hypercalciuria in a Mouse Model Due to a Mutation of the Epithelial Calcium Channel,TRPV5

Hypercalciuria is a major cause of nephrolithiasis, and is a common and complex disorder involving genetic and environmental factors. Identification of genetic factors for monogenic forms of hypercalciuria is hampered by the limited availability of large families, and to facilitate such studies, we screened for hypercalciuria in mice from an N-ethyl-N-nitrosourea mutagenesis programme. We identified a mouse with autosomal dominant hypercalciuria (HCALC1). Linkage studies mapped the Hcalc1 locus to a 11.94 Mb region on chromosome 6 containing the transient receptor potential cation channel, subfamily V, members 5 (Trpv5) and 6 (Trpv6) genes. DNA sequence analysis of coding regions, intron-exon boundaries and promoters of Trpv5 and Trpv6 identified a novel T to C transition in codon 682 of TRPV5, mutating a conserved serine to a proline (S682P). Compared to wild-type littermates, heterozygous (Trpv5 ^682P/+) and homozygous (Trpv5 ^682P/682P) mutant mice had hypercalciuria, polyuria, hyperphosphaturia and a more acidic urine, and ∼10% of males developed tubulointerstitial nephritis. Trpv5 ^682P/682P mice also had normal plasma parathyroid hormone but increased 1,25-dihydroxyvitamin D₃ concentrations without increased bone resorption, consistent with a renal defect for the hypercalciuria. Expression of the S682P mutation in human embryonic kidney cells revealed that TRPV5-S682P-expressing cells had a lower baseline intracellular calcium concentration than wild-type TRPV5-expressing cells, suggesting an altered calcium permeability. Immunohistological studies revealed a selective decrease in TRPV5-expression from the renal distal convoluted tubules of Trpv5 ^682P/+ and Trpv5 ^682P/682P mice consistent with a trafficking defect. In addition, Trpv5^682P/682P mice had a reduction in renal expression of the intracellular calcium-binding protein, calbindin-D_28K, consistent with a specific defect in TRPV5-mediated renal calcium reabsorption. Thus, our findings indicate that the TRPV5 S682P mutant is functionally significant and study of HCALC1, a novel model for autosomal dominant hypercalciuria, may help further our understanding of renal calcium reabsorption and hypercalciuria.     

How aphid alarm pheromone can control aphids: a review

Aphids are the major pests of arable crops, mostly in temperate regions. They are monophagous as well as polyphagous. They inflict damage in brassica, potato, cotton, vegetable and fruit crops. They damage their host plant directly by feeding upon their phloem sap, or indirectly by transmitting pathogens to them. Their life cycle can be autoecious as well as heteroecious. Aphids use semiochemicals for various purposes, in gathering information from their environment and for communication among themselves. They protect themselves from predators and parasitoids by escape response which is arbitrated by use of alarm pheromone signalling. When alarm pheromone, (E)-ß-farnesene, is released, nearby aphids exhibit a variety of behaviours like moving away, running, dropping off the plant and even attacking the predator. Previous studies of integrated pest management strategies have been aimed at the usage of alarm pheromone. However, scientists require complete knowledge of aphid ecology as well as aphid interaction with its natural enemies to establish efficient and viable biological control. This review presents analysis of the existing aphid pest management methodologies and effectiveness of alarm pheromone on aphids and their natural enemies.     

Resistance to Chilo infuscatellus (Lepidoptera: Pyraloidea) in transgenic lines of sugarcane expressing Bacillus thuringiensis derived Vip3A protein

Molecular Biology Reports - Sustainable agriculture requires management of insect pests through resistance development. The biological potential of Cry toxins and Vip protein, derived from Bacillus...