Diosgenin from Dioscorea bulbifera: Novel Hit for Treatment of Type II Diabetes Mellitus with Inhibitory Activity against α-Amylase and α-Glucosidase

Diabetes mellitus is a multifactorial metabolic disease characterized by post-prandial hyperglycemia (PPHG). α-amylase and α-glucosidase inhibitors aim to explore novel therapeutic agents. Herein we report the promises of Dioscorea bulbifera and its bioactive principle, diosgenin as novel α-amylase and α-glucosidase inhibitor. Among petroleum ether, ethyl acetate, methanol and 70% ethanol (v/v) extracts of bulbs of D. bulbifera, ethyl acetate extract showed highest inhibition upto 72.06 ± 0.51% and 82.64 ± 2.32% against α-amylase and α-glucosidase respectively. GC-TOF-MS analysis of ethyl acetate extract indicated presence of high diosgenin content. Diosgenin was isolated and identified by FTIR, ¹H NMR and ¹³C NMR and confirmed by HPLC which showed an α-amylase and α-glucosidase inhibition upto 70.94 ± 1.24% and 81.71 ± 3.39%, respectively. Kinetic studies confirmed the uncompetitive mode of binding of diosgenin to α-amylase indicated by lowering of both Km and Vm. Interaction studies revealed the quenching of intrinsic fluorescence of α-amylase in presence of diosgenin. Similarly, circular dichroism spectrometry showed diminished negative humped peaks at 208 nm and 222 nm. Molecular docking indicated hydrogen bonding between carboxyl group of Asp300, while hydrophobic interactions between Tyr62, Trp58, Trp59, Val163, His305 and Gln63 residues of α-amylase. Diosgenin interacted with two catalytic residues (Asp352 and Glu411) from α-glucosidase. This is the first report of its kind that provides an intense scientific rationale for use of diosgenin as novel drug candidate for type II diabetes mellitus.     

Antioxidant,antibacterial and cytotoxic potential of silver nanoparticles synthesized using terpenes rich extract of Lantana camara L. leaves

Several attempts have been made for green synthesis of silver nanoparticles (AgNPs) using different plant extracts. Present study revealed that, antioxidant, antibacterial and cytotoxic AgNPs were synthesized using terpenes-rich extract (TRE) of environmentally notorious Lantana camara L. leaves. AgNPs were characterized by advanced techniques like UV–Visible and Infra red spectroscopy; XRD, SEM techniques as terpenes coated sphere shaped NPs with average diameter 425 nm. Further, on evaluation, AgNPs were found to exhibit dose – dependent antioxidant potential, good to moderate antibacterial activity against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa; and toxicity on Brine shrimp (A. salinanauplii) with LD₅₀ value 514.50 µg/ml.     

Protective Effect of Selenomethionine on Aflatoxin B1-Induced Oxidative Stress in MDCK Cells

AFB1 is a mycotoxin which exerts their cytotoxicity through increasing oxidative damage in target organ. Kidney is one of target organs vulnerable to damage caused by AFB1. In this study, Madin-Darby canine kidney (MDCK) cells were used to evaluate the AFB1-induced cell damage by the MTT assay. The results revealed that the toxic effect of AFB1 on MDCK cells is both dose and time dependent. Half maximal toxic concentration (IC₅₀) was noted at 0.25 μg/ml of AFB1. Further, protective effect of six different concentrations (0.2, 0.8, 1, 2, 4, and 8 μM) of selenomethionine (SeMet) was observed against 0.25 μg/ml of AFB1-induced damage. The results showed that 0.25 μg/ml of AFB1 caused significant increase in oxidative stress, which was demonstrated by significant increase of malondialdehyde (MDA) level, reduction of intracellular GSH level, as well as GPX1 activity and mRNA level in MDCK cells when compared with control. SeMet protected the cells from AFB1-induced oxidative damage in a dose-dependant manner. Good protection could be achieved between 1 and 4 μM of concentration. Amid this range, MDA level significantly decreased while intracellular GSH level and GPX1 activity in addition to mRNA level significantly increased. Moreover, cell viability was significantly improved. It could be concluded that SeMet is a potential antioxidative agent to alleviate AFB1-induced oxidative stress.     

Dietary Supplementation of Selenium-Enriched Probiotics Enhances Meat Quality of Broiler Chickens (<Emphasis Type="Italic">Gallus gallus domesticus</Emphasis>) Raised Under High Ambient Temperature

We investigated the effects of selenium-enriched probiotics (SP) on broiler meat quality under high ambient temperature and explore their underlying mechanisms. A total of 200 1-day-old male broiler chicks (Ross 308) were randomly allotted to four treatment groups, each with five replicates, in groups of ten birds. These birds were fed a corn-soybean basal diet (C), a basal diet plus probiotics supplementation (P), a basal diet plus Se supplementation in the form of sodium selenite (SS, 0.30 mg Se/kg), and a basal diet with the addition of selenium-enriched probiotics (SP, 0.30 mg Se/kg). The experiment lasted for 42 days. The birds were sacrificed by cervical dislocation, and the breast muscles were removed for further process. Our results showed that SP diet significantly increased (p < 0.05) the physical (pH, colors, water holding capacity, drip loss, shear force) and sensory characteristics of breast meat. All P, SS, and SP supplementation enhanced the antioxidant system by increasing (p < 0.05) the Se concentrations, glutathione (GSH) levels, activities of glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) whereas decreasing (p < 0.05) malondialdehyde (MDA) levels, with SP being higher than P and SS. Moreover, SP diet significantly upregulated (p < 0.05) the mRNA levels of glutathione peroxidase genes (GPx1, GPx4) while it downregulated heat stress biomarkers such as heat shock protein (HSP) 70 as compared to C, P, and SS diets. In conclusion, our findings suggest that SP may function as beneficial nutritive supplement that is capable of improving meat quality during the summer season.     

Detection of measles,mumps and rubella viruses by immuno‐colorimetric assay and its application in focus reduction neutralization tests

Measles, mumps and rubella are vaccine‐preventable diseases; however limited epidemiological data are available from low‐income or developing countries. Thus, it is important to investigate the transmission of these viruses in different geographical regions. In this context, a cell culture‐based rapid and reliable immuno‐colorimetric assay (ICA) was established and its utility studied. Twenty‐three measles, six mumps and six rubella virus isolates and three vaccine strains were studied. Detection by ICA was compared with plaque and RT‐PCR assays. In addition, ICA was used to detect viruses in throat swabs (n = 24) collected from patients with suspected measles or mumps. Similarly, ICA was used in a focus reduction neutralization test (FRNT) and the results compared with those obtained by a commercial IgG enzyme immuno assay. Measles and mumps virus were detected 2 days post‐infection in Vero or Vero‐human signaling lymphocytic activation molecule cells, whereas rubella virus was detected 3 days post‐infection in Vero cells. The blue stained viral foci were visible by the naked eye or through a magnifying glass. In conclusion, ICA was successfully used on 35 virus isolates, three vaccine strains and clinical specimens collected from suspected cases of measles and mumps. Furthermore, an application of ICA in a neutralization test (i.e., FRNT) was documented; this may be useful for sero‐epidemiological, cross‐neutralization and pre/post‐vaccine studies.     

Synthesis, characterization, X-ray structure and DNA photocleavage by cis-dichloro bis(diimine) Co(III) complexes

Complexes of the type [Co(LL)2Cl2]Cl, where LL = N,N'-ethylenediamine (en), 2,2'-bipyridine (bpy), 1,10-phenanthroline (phen), 1,10-phenanthroline-5,6-dione (phendione) and dipyrido[3,2-a:2',3'-c]phenazine (dppz) have been synthesized and characterized by elemental analyses, IR, UV-visible and NMR spectroscopy. Crystal structure of [Co(phendione)2Cl2]Cl x 0.5 HCl x 3.5 H2O has been solved and refined to R = 0.0552. The crystal is monoclinic with space group C2/c; a = 25.730(2) A, b = 12.375(1) A, c = 18.979(2) A, beta = 119.925(1) degrees and Z = 8. The DNA binding characteristics of the complexes, investigated by covalent binding assay, viscosity measurements and competitive binding fluorescence measurements show that the complexes interact with DNA covalently except the complex containing the planar dppz ligand which intercalates within the base pairs of DNA. The complexes containing en, phen and phendione cleave plasmid pBR 322 DNA upon irradiation under aerobic conditions while the complex containing the dppz ligand cleaves DNA upon irradiation under inert atmosphere. Molecular modeling studies show that the minimized structure of [Co(phendione)2Cl2]+, maintained the octahedral structure while binding to the N7 of guanines and the ligand fits into the major groove without disrupting the helical structure of the B-DNA.     

Utility of neutralization test for laboratory diagnosis of suspected mumps

Mumps is an infectious disease caused by mumps virus (MuV), which belongs to the family Paramyxoviridae and genus Rubulavirus. Typical symptoms of mumps include fever and swelling of the parotid glands; however, mumps can be asymptomatic. Mumps is diagnosed by molecular and serological methods (i.e., PCR and Enzyme Immunoassay [EIA]); however, both methods have pros and cons. This study was performed to compare the diagnostic utility of a focus reduction neutralization test (FRNT) to that of MuV‐specific commercial IgM and IgG antibody EIA in patients suspected of having mumps. One hundred‐eighty six samples collected during mumps outbreak in 2012–16 were studied. Samples (n = 80) were tested by all the three serological assays and showed 70.4%, 83% and 92.5% positivity by IgM EIA, IgG and FRNT, respectively. In all, 58.8% samples (n = 47) tested positive in all three assays. Concordance between mumps RT‐PCR and IgM EIA was highest during the first 2–5 days and decreased with increasing time post‐onset. Mumps FRNT results agreed with those of RT‐PCR/IgM EIA from the second week onwards, whereas the results of mumps IgG EIA agreed with those of RT‐PCR/IgM EIA from post‐onset days 3–10. These findings suggest the utility of a FRNT for laboratory diagnosis of mumps in countries whose populations are not immunized against this infection.