A rare case of type II tri-allelic inheritance at vWA, SE33, D8S1179, and D13S317 loci demonstrated by STR analysis in paternity testing.

Short tandem repeat (STR) typing has been regularly used in paternity disputes and forensic human identification linked caseworks. Occasionally, forensic scientists come across aberrant allele patterns during STR typing because of mutations, genetic variations, and other abnormalities. The tri-allelic pattern of STR is rare, particularly, the case where this pattern exists at 4 loci. Here, we report the type II tri-allelic patterns observed at vWA, SE33, D8S1179, and D13S317 loci in the product of conception (POC) sample during the course of our regular paternity case investigation. The DNA extracted from the blood samples and tissue of POC were subjected to STR typing for autosomal and sex STR loci using the commercial QIAGEN's Investigator® IDplex Plus Kit and QIAGEN's Investigator® 24plex QS Kit. Capillary electrophoresis was carried out in 3500 and 3500xL Genetic Analyzer Applied Biosystems and genotyped using GeneMapper ID-X Software v1.5 and v1.6. In this case of paternity inclusion, the POC sample displayed type II tri-allelic patterns at vWA (16, 19, 20), SE33 (19, 28.2, 29.2), D13S317 (16, 19, 20), and D8S1179 (10, 13, 17) loci. In addition, the POC displayed an abnormal genotype with a heterozygous peak imbalance (type II-B) of (1:2) pattern at D3S1358, D21S11, and D16S539 loci, of (2:1) pattern at D1S1656, D12S391, D10S1248, D2S1338, D2S441, D18S317, FGA, CSF1PO, and D5S818 loci, and type II-C allelic pattern (one single peak with triplicate height) at D19S433 and DS7820 loci. Understanding of such anomalous genotypes improves the knowledge about tri-allelic pattern of CODIS loci and helps in the appropriate interpretation of the results in STR typing.

α-Geminal disubstituted pyrrolidine iminosugars and their C-4-fluoro analogues: Synthesis, glycosidase inhibition and molecular docking studies.

A simple strategy for the synthesis of α-geminal disubstituted pyrrolidine iminosugars 3a-c and their C-4 fluorinated derivatives 4a-c has been described from d-glucose. The methodology involves the Corey-Link and Jocic-Reeve reaction with 3-oxo-α-d-glucofuranose and nucleophilic displacement reaction to get the furanose fused pyrrolidine ring skeleton with requisite CH2OH/CO2H functionalities at C-3. The fluorine substituent in target molecules was introduced by nucleophilic displacement of -OTf in 9a/9c with CsF. Appropriate manipulation of the anomeric carbon in the furanose fused pyrrolidine ring skeleton afforded α-geminal disubstituted pyrrolidine iminosugars 3a-c and C-4 fluoro derivatives 4a-c. The pyrrolidine iminosugars 3a and 3c were found to be potent inhibitors of α-galactosidase while, the fluoro derivatives 4a and 4b showed strong inhibition of ß-glucosidase and ß-galactosidase, respectively. These results were substantiated by in silico molecular docking studies.

Iminosugars Spiro-Linked with Morpholine-Fused 1,2,3-Triazole: Synthesis, Conformational Analysis, Glycosidase Inhibitory Activity, Antifungal Assay, and Docking Studies.

Synthesis of iminosugars 1, 2, 3a, and 4a and N-alkyl (ethyl, butyl, hexyl, octyl, decyl, and dodecyl) derivatives 3b-g and 4b-g spiro-linked with morpholine-fused 1,2,3-triazole is described. Conformation of the piperidine ring in each spiro-iminosugar was evaluated by 1H NMR spectroscopy, and conformational change in N-alkylated compounds 4b-g with respect to parent spiro-iminosugar 4a is supported by density functional theory calculations. Out of 16 new spiro-iminosugars, the spiro-iminosugars 3a (IC50 = 0.075 µM) and 4a (IC50 = 0.036 µM) were found to be more potent inhibitors of α-glucosidase than the marketed drug miglitol (IC50 = 0.100 µM). In addition, 3a (minimum inhibition concentration (MIC) = 0.85 µM) and 4a (MIC = 0.025 µM) showed more potent antifungal activity against Candida albicans than antifungal drug amphotericin b (MIC = 1.25 µM). In few cases, the N-alkyl derivatives showed increase of α-glucosidase inhibition and enhancement of antifungal activity compare to the respective parent iminosugar. The biological activities were further substantiated by molecular docking studies.

Azetidine- and N-carboxylic azetidine-iminosugars as amyloglucosidase inhibitors: synthesis, glycosidase inhibitory activity and molecular docking studies.

A simple strategy for the synthesis of hitherto unknown azetidine iminosugars 2a­2c and N-carboxylic azetidine iminosugar 2d has been reported. The methodology involves the conversion of 1,2:5,6-di-O-isopropylidene-3-oxo-α-D-glucofuranose 3 to 3-azido-3-deoxy-3-C-(formyl)-1,2:5,6-di-O-isopropylidene-α-D-glucofuranose 5 using the Jocic­Reeve and Corey­Link approaches. Compound 5 was transformed to 5-OTs 10/5-OMs 19 derivatives that on intramolecular nucleophilic displacement with in situ generated 3-amino functionality afforded the key azetidine ring skeletons 11 and 20, respectively. Hydrolysis of the 1,2-acetonide group and manipulation of the anomeric carbon in 12 provided azetidine iminosugars 2a­2c. In an attempt to synthesize azetidine iminosugars with an additional 4-hydroxymethyl group from 20, we encountered an interesting observation wherein the N-Cbz group in 20 hydrolyzed to the N-COOH functionality under TFA:H2O conditions that gave access for the synthesis of N-carboxylic azetidine iminosugar 2d. The glycosidase inhibitory activity of 2a­2d and intermediates 2e­f was studied with various glycosidases and was compared with Miglitol and 1-deoxynojirimycin (DNJ). Azetidine iminosugars 2 were found to inhibit amyloglucosidase with competitive type inhibition, amongst which 2d was found to be more active than Miglitol and DNJ. These results were substantiated by in silico molecular docking studies.