Relationship, Workload: A Study in Mickey and the Memsahib

Good relationship is an essence for happy and peaceful life as water is necessary for existence of living beings. Extensive workloads and lust for huge earnings compel many professionals, private and government employees to be stuck with their duties for extra hours in the cosmopolitan cities; aloof from their family, relatives and friends; with them they should spend some time to sustain good relationship but they have no time to do so. Contrary, confusion and doubt has penetrate in their minds that disturb many people and force human beings to stay in perturbed circumstances though proper discussion restore their relationship with family particularly. They should spare time at home to share their ups and downs with wife, mother, sisters, brothers and other relatives in the high-tech cities and towns. Therefore, an appreciable relationship has taken a resentful situation when Professor and Memsahib become incompetent and careless to maintain their life-bonding relationship in the Marathi play Mickey and the Memsahib that is written by Satish Alekar (1949-). Professor has been doing research on the mouse named Mickey; so he has a hectic life of taking care of the mouse, scrutinizing research papers that are generally submitted by his research scholars, preparing lectures in the university, maintaining official works because he has been playing role of HOD for years. Memsahib is an Associate Professor who is always busy in her works. They do not negotiate nor give sufficient time to each other that results loneliness, sadness and dilemma

Quantitative determination of alliin in dried garlic cloves and products by high-performance thin-layer chromatography

Purpose: To standardize the garlic samples and its products for alliin contents.Methods: A direct high-performance thin-layer chromatographic (HPTLC) method was developed to determine alliin in Chinese (CG) and Indian garlic (IG) and two other marketed products from USA and UK, respectively. Scanning and quantification were performed at 205 nm. Furthermore, chromatography was performed on pre-coated HPTLC plates with the solvent mixture, n – hexane:ethyl acetate (29:1 v/v), as the mobile phase. In addition, the parameters suggested by International Conference on Harmonization for analytical procedures were considered to validate the proposed method.Results: The system gave a compact spot of alliin at RF = 0.19 ± 0.01 The linear regression data for the calibration plots showed a good linearity (r2 = 0.996) in the concentration range of 200 - 1600 ng. Linear regression equation was represented by Y = 1.792x + 182.855, while LOD and LOQ values were 40.42 ngband-1 and 111.72 ng.band-1, respectively. The method showed excellent accuracy with recovery of 98.20 – 99.10 % and good precision with RSD of 1 - 2.65 %.Conclusion: The proposed method is selective, sensitive and accurate for the determination of alliin in garlic and its products. It is also evident from the results obtained that raw Indian garlic has higher alliin content than Chinese garlic.Keywords: Garlic, HPTLC, Alliin, Hypercholesteremia, Quantificatio

MicroRNA-210-mediated proliferation, survival, and angiogenesis promote cardiac repair post myocardial infarction in rodents

The final publication is available at Springer via http://dx.doi.org/10.1007/s00109-017-1591-8.An innovative approach for cardiac regeneration following injury is to induce endogenous cardiomyocyte (CM) cell cycle re-entry. In the present study, CMs from adult rat hearts were isolated and transfected with cel-miR-67 (control) and rno-miR-210. A significant increase in CM proliferation and mono-nucleation were observed in miR-210 group, in addition to a reduction in CM size, multi-nucleation, and cell death. When compared to control, β-catenin and Bcl-2 were upregulated while APC (adenomatous polyposis coli), p16, and caspase-3 were downregulated in miR-210 group. In silico analysis predicted cell cycle inhibitor, APC, as a direct target of miR-210 in rodents. Moreover, compared to control, a significant increase in CM survival and proliferation were observed with siRNA-mediated inhibition of APC. Furthermore, miR-210 overexpressing C57BL/6 mice (210-TG) were used for short-term ischemia/reperfusion study, revealing smaller cell size, increased mono-nucleation, decreased multi-nucleation, and increased CM proliferation in 210-TG hearts in contrast to wild-type (NTG). Likewise, myocardial infarction (MI) was created in adult mice, echocardiography was performed, and the hearts were harvested for immunohistochemistry and molecular studies. Compared to NTG, 210-TG hearts showed a significant increase in CM proliferation, reduced apoptosis, upregulated angiogenesis, reduced infarct size, and overall improvement in cardiac function following MI. β-catenin, Bcl-2, and VEGF (vascular endothelial growth factor) were upregulated while APC, p16, and caspase-3 were downregulated in 210-TG hearts. Overall, constitutive overexpression of miR-210 rescues heart function following cardiac injury in adult mice via promoting CM proliferation, cell survival, and angiogenesis

Attenuation of methylglyoxal-induced glycation and cellular dysfunction in wound healing by Centella cordifolia

Current pre-clinical evidences of Centella focus on its pharmacological effects on normal wound healing but there are limited studies on the bioactivity of Centella in cellular dysfunction associated with diabetic wounds. Hence we planned to examine the potential of Centella cordifolia in inhibiting methylglyoxal (MGO)-induced extracellular matrix (ECM) glycation and promoting the related cellular functions. A Cell-ECM adhesion assay examined the ECM glycation induced by MGO. Different cell types that contribute to the healing process (fibroblasts, keratinocytes and endothelial cells) were evaluated for their ability to adhere to the glycated ECM. Methanolic extract of Centella species was prepared and partitioned to yield different solvent fractions which were further analysed by high performance liquid chromatography equipped with photodiode array detector (HPLC-PDA) method. Based on the antioxidant [2,2-diphenyl-1-picrylhydrazyl (DPPH) assay] screening, anti-glycation activity and total phenolic content (TPC) of the different Centella species and fractions, the ethyl acetate fraction of C. cordifolia was selected for further investigating its ability to inhibit MGO-induced ECM glycation and promote cellular distribution and adhesion. Out of the three Centella species (C. asiatica, C. cordifolia and C. erecta), the methanolic extract of C. cordifolia showed maximum inhibition of Advanced glycation end products (AGE) fluorescence (20.20 ± 4.69 %, 25.00 ± 3.58 % and 16.18 ± 1.40 %, respectively). Its ethyl acetate fraction was enriched with phenolic compounds (3.91 ± 0.12 mg CAE/μg fraction) and showed strong antioxidant (59.95 ± 7.18 μM TE/μg fraction) and antiglycation activities. Improvement of cells spreading and adhesion of endothelial cells, fibroblasts and keratinocytes was observed for ethyl acetate treated MGO-glycated extracellular matrix. Significant reduction in attachment capacity of EA.hy926 cells seeded on MGO-glycated fibronectin (41.2%) and attachment reduction of NIH3t3 and HaCaT cells seeded on MGO-glycated collagen (33.7% and 24.1%, respectively) were observed. Our findings demonstrate that ethyl acetate fraction of C. cordifolia was effective in attenuating MGO-induced glycation and cellular dysfunction in the in-vitro wound healing models suggesting that C. cordifolia could be a potential candidate for diabetic wound healing. It could be subjected for further isolation of new phytoconstituents having potential diabetic wound healing properties

Pre-Conditioning Stem Cells in a Biomimetic Environment for Enhanced Cardiac Tissue Repair: In Vitro and In Vivo Analysis

Introduction Stem cell-based therapies represent a valid approach to restore cardiac function due to their beneficial effect in reducing scar area formation and promoting angiogenesis. However, their translation into the clinic is limited by the poor differentiation and inability to secrete sufficient therapeutic factors. To address this issue, several strategies such as genetic modification and biophysical pre-conditioning have been used to enhance the efficacy of stem cells for cardiac tissue repair. Methods In this study, a biomimetic approach was used to mimic the natural mechanical stimulation of the myocardium tissue. Specifically, human adipose-derived stem cells (hASCs) were cultured on a thin gelatin methacrylamide (GelMA) hydrogel disc and placed on top of a beating cardiomyocyte layer. qPCR studies and metatranscriptomic analysis of hASCs gene expression were investigated to confirm the correlation between mechanical stimuli and cardiomyogenic differentiation. In vivo intramyocardial delivery of pre-conditioned hASCs was carried out to evaluate their efficacy to restore cardiac function in mice hearts post-myocardial infarction. Results The cyclic strain generated by cardiomyocytes significantly upregulated the expression of both mechanotransduction and cardiomyogenic genes in hASCs as compared to the static control group. The inherent angiogenic secretion profile of hASCs was not hindered by the mechanical stimulation provided by the designed biomimetic system. Finally, in vivo analysis confirmed the regenerative potential of the pre-conditioned hASCs by displaying a significant improvement in cardiac function and enhanced angiogenesis in the peri-infarct region. Conclusion Overall, these findings indicate that cyclic strain provided by the designed biomimetic system is an essential stimulant for hASCs cardiomyogenic differentiation, and therefore can be a potential solution to improve stem-cell based efficacy for cardiovascular repair

Isolation, biological evaluation and validated HPTLC-quantification of the marker constituent of the edible Saudi plant Sisymbrium irio L.

AbstractPhytochemical investigation and chromatographic purification of the n-hexane fraction of the aerial parts of the edible Saudi plant Sisymbrium irio led to the isolation of β-sitosterol (1), stigmasterol (2) and β-sitosterol-β-d-glucoside (3). The cytotoxic effects of the n-hexane, dichloromethane, ethyl acetate and n-butanol fractions were tested against three cancer cell lines viz., MCF-7, HCT-116 and HepG2, using the crystal violet staining (CVS) method, while the antibacterial activity against a number of pathogenic bacterial strains, was also estimated using the broth microdilution assay. The n-hexane fraction showed potent cytotoxic activities against all tested human cancer cell lines (IC50: 11.7–13.4μg/mL), while the dichloromethane fraction was particularly potent against HCT-116 cells (IC50: 5.42μg/mL). On the other hand, the n-hexane and EtOAc fractions demonstrated significant inhibitory activities against the Gram positive bacteria S. pyogenes and C. perfringens; and the Gram negative bacterium S. enteritidis. Our results warrant the therapeutic potential of S. irio as nutritional supplement to reduce the risk of contemporary diseases. Additionally, a validated high performance thin-layer chromatography (HPTLC) method was developed for the quantitative analysis of biomarker β-sitosterol glucoside (isolated in high quantity) from the n-hexane fraction. The system was found to furnish a compact, sharp, symmetrical and high resolution band for β-sitosterol glucoside (Rf=0.43±0.002). The limit of detection (LOD) and limit of quantification (LOQ) for β-sitosterol glucoside was found to be 21.84 and 66.18ngband−1, respectively. β-sitosterol glucoside was found to be present only in n-hexane fraction (2.10μg/mg of dried fraction) while it was absent in the other fractions of S. irio which validated the high cytotoxic and antibacterial activity of n-hexane fraction of S. irio

A NOVEL ACYCLIC DITERPENIC ALCOHOL ISOLATED FROM ANTIOXIDANT ACTIVE ETHANOL EXTRACT OF LEAVES OF CENTAUROTHAMNUS MAXIMUS GROWN IN SAUDI ARABIA

Background: Isolation and characterization of a new compound from the antioxidant active ethanol extract of leaves of an endemic plant Centaurothamnus maximus. Methods: The air dried powdered leaves of the plant was extracted successively with n-hexane, dichloromethane, ethyl acetate and ethanol. The obtained extracts were concentrated under reduced pressure using rotary evaporator. The antioxidant activity was carried out on various concentrations (1000, 500, 100, 50 and 10 µg/ml) of all the extracts by DPPH free radical scavenging method. After screening for antioxidant potential the ethanol fraction was selected for the isolation of phytoconstituents by column chromatography using LiChroprep RP-18 as stationary phase and water, MeOH and CHCl3 in different combinations as eluent. The chemical structures of the isolated compounds were elucidated by 1D and 2D NMR spectroscopic techniques (DEPT, COSY, H M B C and HSQC) aided by EIMS mass and IR spectra. Results: The antioxidant activity of ethanol extract was highly comparable with standard ascorbic acid as compared to other extracts. A new compound along with two known compounds has been isolated from antioxidant active ethanol extract. The chemical structure of unknown compound was established as 2, 6, 10, 14-tetramethyl hexadec-12-cis-en-5α, 7α, 9α, 14α-tetraol (CM1) while the known compound isolated Luteolin-7-O-β-glucopyranosyl-6''-O-(6''→1''')-β-D-rhamno-pyranoside (CM-2) and stigmast-5,22–dien-3β –ol (CM3). Conclusion: On the basis of interpretation of different spectroscopy data we concluded that the compound CM1 is an acyclic diterpenic alcohol. The authors are reporting the isolation of CM1 from plant source for the first time but CM2 and CM3 are known compounds. Ethanol extract of leaves can be recommended as potent antioxidant for ethno-medical purposes. The antioxidant properties might be due to some well-known antioxidants like (CM-2) and other falvonoidal compounds

COMPARATIVE PROFILING OF BIOMARKER PSORALEN IN ANTIOXIDANT ACTIVE EXTRACTS OF DIFFERENT SPECIES OF GENUS FICUS BY VALIDATED HPTLC METHOD

Background: A simple but sensitive HPTLC method was developed for the comparative evaluation of psoralen in antioxidant active extracts of leaves of five different species of genus Ficus (Ficus carica, Ficus nitida, Ficus ingens, Ficus palmata and Ficus vasta). Materials and Methods: HPTLC studies were carried out using CAMAG HPTLC system on Glass-backed silica gel 60F254 HPTLC pre-coated plates using selected mobile phase toluene: methanol (9:1). The antioxidant activity was carried out, using DPPH free radical method. Results: Among all the five species of genus Ficus, F. palmata and F. carica exhibited comparatively good antioxidant activity in DPPH assay. The developed HPTLC method was found to give a compact spot for psoralen (Rf = 0.55±0.001) at 305 nm. The regression equation and r2 for psoralen was found to be Y= 4.516X+35.894 and 0.998. The quantification result revealed the presence of psoralen in only two species, F. carica (0.24%, w/w) and F. palmata (1.88%, w/w) which supported their supremacy for anti-oxidant potential over other species. The statistical analysis proved that the developed method was reproducible and selective. Conclusion: The developed method can be used as an important tool to assure the therapeutic dose of active ingredients in herbal formulations as well as for standardization and quality control of bulk drugs and in-process formulations. This method can also be employed for the further study of degradation kinetics and determination of psoralen in plasma and other biological fluids

Pharmacological Evaluation of Secondary Metabolites and Their Simultaneous Determination in the Arabian Medicinal Plant Plicosepalus curviflorus Using HPTLC Validated Method

© The Author(s) 2019. The present study aimed to identify biologically active secondary metabolites from the rare plant species, Pulsatilla patens subsp. patens and the cultivated P. vulgaris subsp. vulgaris. Chromatographic fractionation of the ethanolic extract of the roots of P. patens subsp. patens resulted in the isolation of two oleanane-type glycosides identified as hederagenin 3-O-β-d-glucopyranoside (2.7 mg) and hederagenin 3-O-β-d-galactopyranosyl-(1→2)-β-d-glucopyranoside (3.3 mg, patensin). HPLC analysis of the methanolic extract of the crude root of P. patens subsp. patens and P. vulgaris subsp. vulgaris revealed the presence of Pulsatilla saponin D (hederagenin 3-O-α-l-rhamnopyranosyl(1→2)-[β-d-glucopyranosyl(1→4)]-α-l-arabinopyranoside). Chromatographic analysis using GC-MS of the silylated methanolic extracts from the leaves and roots of these species identified the presence of carboxylic acids, such as benzoic, caffeic, malic, and succinic acids. The extracts from Pulsatilla species were tested for their antifungal, antimicrobial, and antimalarial activities, and cytotoxicity to mammalian cell lines. Both P. patens subsp. patens and P. vulgaris subsp. vulgaris were active against the fungus Candida glabrata with the half-maximal inhibitory concentration (IC 50 ) values of 9.37 µg/mL and 11 µg/mL, respectively. The IC 50 values for cytotoxicity evaluation were in the range of 32–38 μg/mL for P. patens subsp. patens and 35–57 μg/mL for P. vulgaris subsp. vulgaris for each cell line, indicating general cytotoxic activity throughout the panel of evaluated cancer and noncancer cells

Phytochemical investigation of Calotropis procera Ait roots

443-446Phytochemical investigation of the roots of Calotropis procera Ait. (Asclepiadaceae) yields two new phytoconstituents procerursenyl acetate and proceranol together with the known compounds N-dotriacont-6-ene, glyceryl mono-oleolyl-2-phosphate, methyl myrisate, methyl behenate and glyceryl-1,2-dicapriate-3-phosphate. The structures of the new compounds have been identified as urs-18α-H-12,20(30)-diene-3β-yl acetate and n-triacontan -10β-ol on the basis of spectral data analysis and chemical reactions