Introduction:Alzheimer's disease (AD) is a progressive neurologic disease of the brain that affects intellectual abilities, reasoning and memory. Acetylcholine (ACh) is involved in the maintenance of cognitive process. Pathologically, ACh production is compromised in the brains of AD affected people. Presence of acetylcholinesterase (AChE) in the synaptic cleft, which hydrolyzes ACh, further decreases the ACh-levels, and thereby, additionally compromises cognition. The tribal people of North East India have been using indigenous plants as traditional medicine for brain disorders. We assayed whether the plants used in the traditional tribal knowledge for the treatment of brain disorders might contain better AChE-inhibitors. Methods: We collected 10 traditional medicinal plants from Northeast India. A total of 39 plant extracts were prepared using three solvent systems. The Acetylcholinesterase (AChE) activity was measured with Ellman method. The experiment was done in triplicate for each level of inhibitor. The activity was measured at 412 nm wavelength using Plate Reader. The standard student t-test was used to show significant difference in IC50 values between extracts. Results: The result are reported based on Km, Vmax, IC 50 (µg/µl), percentage inhibition and inhibition pattern. Two extracts had competitive inhibition, 11 extracts had mixed inhibition, 2 extracts had non-competitive inhibition, 11 extracts had uncompetitive inhibition and 4 extracts did not provide any proper pattern. The IC 50 for these plant extracts were at the range of 0.51-12.4 µg/µl. Notably, Cinnamomum camphora (leaf: chloroform), Litsea glutinosa (stem; chloroform), and Litsea glutinosa (stem; methanol) showed IC 50 values of 0.51, 0.53 & 0.81 µg/µl, respectively.

SummaryObjectiveJuvenile myoclonic epilepsy (JME) is the most common form of idiopathic generalized epilepsies (IGEs) and is genetically heterogeneous. Mutations in EFHC1 cause JME. Because about 2 million people in India are affected by JME alone, we investigated the prevalence of mutations in the EFHC1 gene in the Indian population with JME. We studied 63 patients with JME and 80 healthy controls.MethodsClinical identification of JME was evaluated using established criteria. Following clinical evaluation of the patients and confirming presence of JME, blood samples were collected from each patient and healthy individual. Subsequently, genomic DNA was extracted from the blood samples. Eleven exons of the EFHC1 gene were individually amplified by polymerase chain reaction (PCR) for each DNA sample. The PCR products were then purified and sequenced commercially. The identified DNA variants were sequenced at least twice in both the forward and reverse directions and compared with the Exome Aggregation Consortium (ExAC) database.ResultsWe found five heterozygous and one homozygous variant. We found three novel coding variants 661C→T, 779 G →A, and 730 C→T, which lead to R221C, R260Q, and R244STOP amino acid substitutions, respectively. The coding variant 475 C→T, resulting in the amino acid substitution R159W, reported earlier as polymorphism, was also identified in both patient and control populations.SignificanceDetection of these three novel variants, excluding R159W, which is considered polymorphism, expands the range of possible mutations in the EFHC1 gene. The novel variants that we are reporting herein have not been mentioned before as occurring in JME patients of other ethnic population. Therefore, these novel coding variants may be confined to the Indian JME population. Further studies on the mutational spectrum of EFHC1 in a larger number of Indian JME patients concurrent with their mode of inheritance and underlying functional assays should establish whether EFHC1 could be a panethnic gene for JME.

No abstract