Hippo effector, Yorkie, is a tumor suppressor in select Drosophila squamous epithelia.

Mammalian Yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ) and Drosophila Yorkie (Yki) are transcription cofactors of the highly conserved Hippo signaling pathway. It has been long assumed that the YAP/TAZ/Yki signaling drives cell proliferation during organ growth. However, its instructive role in regulating developmentally programmed organ growth, if any, remains elusive. Out-of-context gain of YAP/TAZ/Yki signaling often turns oncogenic. Paradoxically, mechanically strained, and differentiated squamous epithelia display developmentally programmed constitutive nuclear YAP/TAZ/Yki signaling. The unknown, therefore, is how a growth-promoting YAP/TAZ/Yki signaling restricts proliferation in differentiated squamous epithelia. Here, we show that reminiscent of a tumor suppressor, Yki negatively regulates the cell growth-promoting PI3K/Akt/TOR signaling in the squamous epithelia of Drosophila tubular organs. Thus, downregulation of Yki signaling in the squamous epithelium of the adult male accessory gland (MAG) up-regulates PI3K/Akt/TOR signaling, inducing cell hypertrophy, exit from their cell cycle arrest, and, finally, culminating in squamous cell carcinoma (SCC). Thus, blocking PI3K/Akt/TOR signaling arrests Yki loss-induced MAG-SCC. Further, MAG-SCCs, like other lethal carcinomas, secrete a cachectin, Impl2-the Drosophila homolog of mammalian IGFBP7-inducing cachexia and shortening the lifespan of adult males. Moreover, in the squamous epithelium of other tubular organs, like the dorsal trunk of larval tracheal airways or adult Malpighian tubules, downregulation of Yki signaling triggers PI3K/Akt/TOR-induced cell hypertrophy. Our results reveal that Yki signaling plays an instructive, antiproliferative role in the squamous epithelia of tubular organs.

Unravelling the key steps impairing the metabolic state of Xanthomonas cells undergoing programmed cell death.

Programmed cell death (PCD) has been reported in Xanthomonas axonopodis pv. glycines (Xag) wild type earlier and was indirectly shown to be induced by metabolic stress; however, deciphering the key proteins regulating the metabolic stress remained unrevealed. In this study, transcriptomic and proteomic analyses were performed to investigate the prominent pathways, having a role in the induction of metabolic stress in Xag cells undergoing PCD. A comprehensive analysis of transcriptome and proteome data revealed the major involvement of metabolic pathways related to branched chain amino acid degradation, such as acyl-CoA dehydrogenase and energy-yielding, ubiquinol:cytochrome c oxidoreductase complex, in Xag cells undergoing PCD. Consequently, oxidative stress response genes showed major upregulation in Xag cells in PCD-inducing medium; however, no such upregulation was observed at the protein level, indicative of depleted protein levels under excessive stress conditions. Activation of stress response and DNA repair proteins was also observed in Xag cells grown in PCD-inducing medium, which is indicative of excessive cellular damage. Thus, the findings indicate that programmed cell death in Xag is an outcome of metabolic stress in nutrient condition not suitable for a plant pathogen like Xanthomonas, which is more acclimatised with altogether a different nutritional requirement predominantly having an enriched carbohydrate source.

Simulation study to assess the effectiveness of gamma radiation for inactivation of viruses on food packaging material.

The recent COVID-19 pandemic spread across the globe has raised the concern about the possible transmission of viruses through food packaging material during domestic and international trade. Therefore, mitigation strategies are needed to address these safety issues. Preliminary in-silico study showed that interactions between food packaging material and viral surface proteins were possibly hydrophobic in nature with most favourable interaction having a binding free energy of -5.24 kcal/mol. Since these interactions can cause viruses to adsorb on the food packets and get transmitted during supply chain, it is necessary to inactivate the viruses. In this context, efficacy of gamma irradiation in inactivating the viruses on the food packaging material was assessed. For this simulation study P1 (virulent) bacteriophage of E. coli was used as a model system. Gamma irradiation of food packets at an absorbed dose >8 kGy was found to completely inactivate the infectivity of P1(virulent) bacteriophage when co-cultured with E. coli host and assayed for viral plaque formation. Reduction in infectivity of P1(vir) phage was more prominent at ambient temperature (25 ± 2 °C) as compared to cold temperature (6 ± 2 °C) when assayed after storage (one week). Gamma irradiation (2 kGy) completely inactivated the virus particles on food packets when stored for 1 week at both the above temperatures. It is thus proposed that gamma irradiation (2 kGy) can possibly be integrated as a final treatment of the packaged food products to rule out the possibility of viral transmission. However, the efficacy of radiation processing against different pathogenic viruses needs to be determined prior to actual commercial deployment.

Control of post-harvest storage losses in water chestnut (Trapa bispinosa Roxburg) fruits by natural functional herbal coating and gamma radiation processing.

Water chestnut (Trapa bispinosa Roxburg) has short shelf life even after drying due to insect infestations resulting in huge economic losses to farmers. Study was performed including coating with Aloe-vera gel, air drying, LDPE packaging and gamma irradiation (≤ 1 kGy) were evaluated for their efficacy in controlling storage losses. The study was performed with complete randomized design up to 6-month storage period. Physico-chemical properties analysis indicated, weight loss (0.11-0.88%), damage (upto 6.5%) and lesser grain borer infestation (up to 16 no. per packet) during storage at different intervals. Moisture content ranged (9.25-10.10%), protein (8.82-8.89%), fat (0.90-1.00%), carbohydrate (76.83-76.89%), total ash (3.11-4.18%), total sugar (5.83-5.89%), reducing sugar (1.84-1.88%), non-reducing sugar (3.98-4.02%), amylose (66.54-66.64%), in-vitro starch digestibility (3.14-3.18%), phenolics (0.14-0.18%), acidity (0.11-0.12%) and ascorbic acid (41.66-50.00 mg/100 g). These treatments were not found to affect the quality attributes significantly however; it helps in controlling insect infestations at ambient condition. The sensory score of the product, 'Sev' was ranked as like slightly to moderately range. It could be concluded that water chestnut with combinations of 50% aloe-vera gel +0.75 kGy gamma radiation treatment could be stored up to 6 months with retention of quality attributes.

Secondary product from strawberry (Fragaria ananassa) fruit for extended preservation and value addition.

Strawberry is highly delicate and perishable fruit prone to microbial spoilage. To address these issues, secondary product(s) using strawberry pulp was prepared by adding sweetener (jaggery or sucrose) and anti-browning agent (citrate) to reduce the extent of drying induced sourness and discoloration, respectively. Subsequently, these products were LDPE packed and radiation processed (≤ 5 kGy) to ensure microbiological safety during ambient temperature storage. Physical, biochemical, functional (antioxidant and antimutagenic) and organoleptic properties (including flavor compounds) were found to be either retained or enhanced compared to dehydrated strawberry pulp. The product termed as dehydrated strawberry roll having jaggery and citrate (DSRJ) was found to be more acceptable than that having sucrose and citrate (DSRS) till 5 months. Scanning electron microscopy indicated better surface quality of DSRJ than DSRS. Electrochemical property when analyzed using cyclic voltammetry displayed strong correlation to radical scavenging antioxidant assays with marginal storage associated shift in the peak potential. GC-MS analysis indicated retention of most of the flavor compounds in DSRJ during storage. Thus, DSRJ, a value-added product could provide a solution to control post-harvest losses of the fruit by processing to a more stable form and ensuring availability beyond the season. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13197-021-05171-9.

No induced mutagenesis in human lymphoblast cell line and bacterial systems upon their prolonged sub-culturing in irradiated food blended media.

BACKGROUND: Profound apprehension towards safety of irradiated food has remained a major cause behind tardy acceptance of this technology although it has immense socio-economic potential. Generation of in-depth scientific evidence will help to refute these apprehensions. With this prospective, the present study was undertaken where safety of various irradiated (Dmin up to 25 kGy) foods was evaluated through long-term exposure studies in models including human lymphoblast TK6 cell line (100 generations) and Escherichia coli MG1655 cells (exclusive sub-culturing in irradiated food medium for 3000 generations). Additionally, the Ames test, micronucleus test, comet assay, DNA sequencing and restriction profiling of phagemid DNA from E. coli cells sub-cultured in irradiated food medium were also performed. RESULTS: No induced mutagenesis was observed in these cells during long-term sub-culturing in various irradiated food medium. Also no change was observed in profiles of comet, micronucleus, restriction digestion, random amplification of polymorphic DNA as well as DNA sequences. The latter also ruled out the possibility of any silent mutation. CONCLUSION: Findings of the current study thus provided credible molecular evidence supporting the safety of irradiated foods. This would be helpful in confidence building among consumers, entrepreneurs, and strengthening the overall food irradiation program to achieve 'food safety' and 'security'. © 2017 Society of Chemical Industry.

Food Processing by Irradiation­­An effective technology for food safety and security.

Radiation processing of food involves controlled application of energy from ionizing radiations from radioisotopes (Cobalt-60 and Caesium-137), electron beam (≤10 MeV) or X-rays (≤5 MeV) in an irradiation chamber shielded by 1.5 - 1.8 m thick concrete walls. Food, either pre-packed or in-bulk, placed in suitable containers is sent into the chamber through an automatic conveyor. Major benefits achieved by radiation processing of food are inhibition of sprouting of tubers and bulbs, delay in ripening and senescence of fruits and vegetables, disinfestations of insect pests in agricultural commodities, destruction of microbes responsible for food spoilage, and elimination of food pathogens and parasites of public health importance. Irradiation produces very little chemical changes in food, and the changes are similar to those by other preservation methods like heat. The radiolytic products and free radicals produced are identical to those present in foods subjected to treatments such as cooking and canning. None of the changes known to occur have been found to be harmful. Radiation processing of food has been approved by various international statutory bodies and organizations to ensure 'Food Security & Safety', and overcome 'Technical barrier to International Trade' and currently is being practiced in more than 60 countries worldwide.

Differential gene expression in irradiated potato tubers contributed to sprout inhibition and quality retention during a commercial scale storage.

Current study is the first ever storage cum market trial of radiation processed (28 tons) of potato conducted in India at a commercial scale. The objective was to affirm the efficacy of very low dose of gamma radiation processing of potato for extended storage with retained quality and to understand the plausible mechanism at the gene modulation level for suppression of potato sprouting. Genes pertaining to abscisic acid (ABA) biosynthesis were upregulated whereas its catabolism was downregulated in irradiated potatoes. Additionally, genes related to auxin buildup were downregulated in irradiated potatoes. The change in the endogenous phytohormone contents in irradiated potato with respect to the control were found to be correlated well with the differential expression level of certain related genes. Irradiated potatoes showed retention of processing attributes including cooking and chip-making qualities, which could be attributed to the elevated expression of invertase inhibitor in these tubers. Further, quality retention in radiation treated potatoes may also be related to inhibition in the physiological changes due to sprout inhibition. Ecological and economical analysis of national and global data showed that successful adoption of radiation processing may gradually replace sprout suppressants like isopropyl N-(3-chlorophenyl) carbamate (CIPC), known to leave residue in the commodity, stabilize the wholesale annual market price, and provide a boost to the industries involved in product manufacturing.

Alpha-cadinol as a potential ACE-inhibitory volatile compound identified from Phaseolus vulgaris L. through in vitro and in silico analysis.

Hypertension is a major risk factor of cardiovascular diseases, which is mainly caused due to over activation of renin-angiotensin system. The angiotensin converting enzyme (ACE), which is involved in formation of angiotensin II from angiotensin I, causes the blood vessels to constrict, in turn leading to hypertension. The current study was initiated to understand the role of bioactive volatile compounds from Phaseolus vulgaris L. (common bean), in ACE enzyme inhibition. Beans aqueous extract (BAE) showed maximum ACE inhibition of 88.4 ± 0.8% in comparison to other commonly consumed vegetables like spinach and garlic. The head space gas chromatography-mass spectrometry analysis showed the presence of a number of terpenes and terpenoids, which were present prominently in BAE. In silico molecular docking studies indicated that among the other volatile compounds, alpha-cadinol (-7.27 kcal/mol) and ar-tumerone (-6.44 kcal/mol) have the maximum binding affinity with the active site of ACE, as compared to that of captopril (-6.41 kcal/mol). The molecular dynamic simulation in biological environment, showed that alpha-cadinol forms a stable complex with ACE, with average binding energy of -42 kJ/mol. The ACE:alpha-cadinol complex was found to be stable mainly due to the hydrophobic interactions of alpha-cadinol with active site residues (Tyr523 and Phe457) of ACE. The in silico drug-likeness analysis showed that alpha-cadinol is appropriate for human system with no predicted hepatotoxicity or mutagenicity (AMES toxicity).Communicated by Ramaswamy H. Sarma.

Comprehensive in situ and ex situ ß-glucosidase-assisted assessment reveals Indian mangoes as reservoirs of glycosidic aroma precursors.

Mango, a valued commercial fruit in India is popular mostly because of its attractive flavour. Glycosidically bound volatiles (GBV), an underrepresented warehouse of aroma, remain completely unexplored in Indian mangoes. In this study, GBV were profiled in pulps and peels of 10 Indian mango cultivars, leading to detection of 66 GBV which were dominated by monoterpenoids and phenolics. Peels were quantitatively and qualitatively richer in GBV than pulps. Hierarchical clustering and principal component analysis indicated higher contribution of peel GBV to the distinctness of cultivars. Linalool, geraniol, and eugenol were the significant contributors based on the odour units. Direct ß-glucosidase treatment to the juice resulted in the release of lesser number of volatiles than those released from the purified GBV extracts. Apart from providing a comprehensive catalogue of GBV in mangoes, our data suggests the need of critical assessment of the usefulness of ß-glucosidases in aroma improvement of fruit juices.

Anti-proliferative effect and underlying mechanism of ethoxy-substituted phylloquinone (vitamin K1 derivative) from Spinacia oleracea leaf and enhancement of its extractability using radiation technology.

In our previous studies, a novel antimutagenic compound, 2-ethoxy-3-(3,7,11,15-tetramethylhexadec-2-ethyl) naphthaquinone-1,4-dione (ethoxy-substituted phylloquinone; ESP) from spinach was characterized and mechanism contributing to its antimutagenicity was deduced. In the current study, anti-proliferative activity of ESP was assessed in lung cancer (A549) cells using MTT [3-(4,5-dimethylthiazole-2yl)-2,5-diphenyl tetrazolium bromide], clonogenic assays and cell cycle analysis. ESP treatment showed selective cytotoxicity against lung cancer cells and no cytotoxicity in normal lung (WI38) cells. Cell cycle analysis revealed that ESP treatment arrests A549 cell population in G2-M phase. In-silico analysis indicated positive drug-likeness features of ESP. Molecular docking showed H-bonding and hydrophobic interactions between ESP and B-DNA dodecamer residues at minor groove. SWATH-MS (Sequential Window Acquisition of All Theoretical Mass Spectra) based proteomic analysis indicated down-regulation of proteins involved in EGFR signaling, NEDDylation and other metabolic pathways and up-regulation of tumor suppressor (STAT1 and NDRG1) proteins. Treatment of spinach powder with gamma radiation (5-20 kGy) from cobalt (Co-60) enhanced the extractability of ESP up to 4.4-fold at the highest dose of 20 kGy. Scanning electron microscopy of spinach powder displayed decrease in smoothness and compactness with increase in radiation dose attributing to its enhanced extractability. Increase in the extractability of ESP with increasing radiation doses as measured by fluorescence intensity and dry weight basis was strongly correlated. Nonetheless, radiation treatment did not affect the functionality of ESP in terms of anti-proliferative and antimutagenic activities. Current findings thus highlight broad spectrum bioactivity of ESP from spinach, its underlying mechanism and applicability of radiation technology in enhancing extractability. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03264-6.

Unconventional ß-Glucosidases: A Promising Biocatalyst for Industrial Biotechnology.

ß-Glucosidases primarily catalyze removal of terminal glucosyl residues from a variety of glucoconjugates and also perform transglycosylation and reverse hydrolysis. These catalytic properties can be readily exploited for degradation of lignocellulosic biomass as well as for pharmaceutical, food and flavor industries. ß-Glucosidases have been either isolated in the native form from the producer organism or recombinantly expressed and gaged for their biochemical properties and substrate specificities. Although almond and Aspergillus niger have been instantly recognizable sources of ß-glucosidases utilized for various applications, an intricate pool of novel ß-glucosidases from different sources can provide their potent replacements. Moreover, one can envisage the better efficacy of these novel candidates in biofuel and biorefinery industries facilitating efficient degradation of biomass. This article reviews properties of the novel ß-glucosidases such as glucose tolerance and activation, substrate specificity, and thermostability which can be useful for their applications in lignocellulose degradation, food industry, and pharmaceutical industry in comparison with the ß-glucosidases from the conventional sources. Such ß-glucosidases have potential for encouraging white biotechnology.

Molecular mechanism of antimutagenicity by an ethoxy-substituted phylloquinone (vitamin K1 derivative) from spinach (Spinacea oleracea L.).

In our previous study, an antimutagenic compound from spinach (Spinacea oleracea L.), ethoxy-substituted phylloquinone (ESP) was isolated and characterized. The current study deals with elucidation of the possible mechanism of antimutagenicity of ESP against ethyl methanesulfonate (EMS) deploying model systems such as human lymphoblast (TK+/- or TK6) cell line (thymidine kinase gene mutation assay) and Escherichia coli MG1655 (rifampicin resistance assay). Findings of the study ruled out the possibility of direct inactivation of EMS by ESP. DAPI competitive binding assay indicated the DNA minor groove binding activity of ESP. Interestingly, ESP did not display major groove binding or intercalating abilities. Further, proteomics study using 2-D gel electrophoresis in E. coli and subsequent studies involving single gene knockout strains revealed the possible role of tnaA (tryptophanase) and dgcP (diguanylate cyclase) genes in observed antimutagenicity. These genes have been reported to be involved in indole and cyclic-di-GMP biosynthesis, respectively, which eventually lead to cell division inhibition. In case of TK+/- cell line system, ADCY genes (adenylate cyclase), a functional analogue of dgcP gene, were found to be transcriptionally up-regulated. The generation/doubling time were significantly higher in E. coli or TK+/- cells treated with ESP than control cells. The findings indicated inhibition of cell proliferation by ESP through gene regulation as a possible mechanism of antimutagenicity across the biological system. Cell division inhibition actually provides additional time for the repair of damaged DNA leading to antimutagenicity.

GC-MS olfactometric characterization of odor active compounds in cooked red kidney beans (Phaseolus vulgaris).

Red kidney beans are a staple pulse crop well known for its unique flavor characterized by kidney bean like, smoky, sulfury and earthy aroma notes. However, nature of compounds responsible for the unique beany odor of the cooked pulse has not been established. Steam distillation extracts of red kidney beans were subjected to Gas-chromatography olfactometry (GC-O) techniques namely detection frequency and aroma extract dilution analysis (AEDA). GC-O results suggest that methional with flavor dilution (FD) factor 21 is responsible for imparting the characteristic odor of the cooked red kidney beans. Apart from this p-vinyl guaiacol (FD 13) was identified as most important contributor towards smoky odor note. Sulfury note was mainly contributed by diethyl sulfide (FD 10) while 2-ethyl-3-methyl pyrazine (FD 13) was identified to be responsible for earthy note in cooked red kidney beans. Contribution of these compounds in characteristic aroma of cooked red kidney beans is reported here for the first time.

Methylation of guar gum for improving mechanical and barrier properties of biodegradable packaging films.

Improving functional properties of biopolymers for use as environment friendly packaging is an area of current interest. Biodegradable films with improved barrier and mechanical properties were prepared from methylated guar gum. Methylation resulted in structural modification of guar gum (GG) promoting greater crystallization thereby enhancing thermal stability towards decomposition. Reduction in radius of gyration (Rg), weight average molecular weight (Mw), and an increase in polydispersity index (PDI) were also observed due to methylation. Methylated guar gum (MGG) films exhibited 40% lower water vapor transmission rate (WVTR) as compared to control purified guar gum (PGG) films. Films prepared by partial replacement of PGG with MGG (10, 25, 50, 75 and 100% w/w) showed gradual improvement in percent elongation. The study gives an insight on the role of methylation in enhancing barrier and mechanical properties of GG based biodegradable films for possible application in food packaging.

Neuroprotective Activity of Evolvulus alsinoides & Centella asiatica Ethanolic Extracts in Scopolamine-Induced Amnesia in Swiss Albino Mice.

AIM: To carry out the comparative nootropic, neuroprotective potentials of two medicinal plant species. MATERIAL AND METHODS: For neuroprotective activity; behavior models (elevated plus maze & morris water maze), in vivo antioxidant (superoxide dismutase, catalase, lipid peroxidation & reduced glutathione), inflammatory markers (IL-1ß, IL-6 & TNF-α) and acetylcholine esterase (AChE) assessment procedures followed at different dosages i.e. 250 & 500 mg/kg of Evolvulus alsinoides and Centella asiatica ethanolic extracts. At the end of the study, it was performed histopathological analysis of the following organs: brain, heart, liver, and kidney. RESULTS: In oral administration of different doses of ethanolic extracts of both medicinal plants i.e. Sco + EEA 250 = 2.49 ± 0.29 , Sco + EEA 500 = 2.67 ± 0.36, Sco + ECA 250 = 2.33 ± 0.17, Sco + ECA 500 = 2.77 ± 0.21, Sco + EEA + ECA 250 = 2.61 ± 0.32 and Sco + EEA + ECA 500 = 2.79 ± 0.16 U/mg of protein respectively against the scopolamine induced group Sco (control) = 5.51 ± 0.35 U/mg of protein extracts shows neuroprotective and nootropic activity with reducing AChE level in the brain homogenate of swiss albino mice. CONCLUSION: Since the E. alsinoides & C. asiatica are already used in traditional Indian medicine as the neuroprotective agent and also found promising effects over inflammatory diseases, wound healing, and immunomodulatory activity. The neuroprotective effect of both plants extracts attributed to inhibition of AChE activity and improve the spatial memory formation.

In Vivo Toxicity Study of Ethanolic Extracts of Evolvulus alsinoides & Centella asiatica in Swiss Albino Mice.

AIM: We aimed to investigate several parameters after the in vivo acute and sub-acute administration of ethanolic extracts from E. alsinoides & C. asiatica. METHODS: Malignant Ovarian Germ Cell Tumors for in vivo toxicity study guidelines 423 and 407 of Organization for Economic Co-operation and Development (OECD) were followed for acute and sub-acute toxicity assays respectively. For LD50 evaluation, a single dose of ethanolic extracts of Evolvulus alsinoides L. (EEA) and ethanolic extracts of Centella asiatica (ECA) was orally administered to mice at doses of 200, 400, 800, 1600 and 2000 mg/kg. Then the animals were observed for 72 hours. For acute toxicity evaluation, a single dose of both extracts was orally administered to mice at doses of 300, 600, 1200 and 2000 mg/kg and the animals were observed for 14 days. In the sub-acute study, the extracts were orally administered to mice for 28 days at doses of 300, 600, 1200 and 2000 mg/kg. To assess the toxicological effects, animals were closely observed on general behaviour, clinical signs of toxicity, body weight, food and water intake. At the end of the study, it was performed biochemical and hematological evaluations, as well as histopathological analysis from the following organs: brain, heart, liver, and kidney. RESULTS: The oral administration of E. alsinoides and C. asiatica ethanolic extracts, i.e. EEA 300, EEA 600, EEA 1200, EEA 2000, ECA 300, ECA 600, ECA 1200 & ECA 2000 mg/kg doses showed no moral toxicity effect in LD50, acute and sub-acute toxicity parameters. CONCLUSION: In this study, we had found that E. alsinoides & C. asiatica extract at different doses cause no mortality in acute and sub-acute toxicity study. Also, histopathology of kidney, liver, heart, and brain showed no alterations in tissues morphology.

Characterization and evaluation of mycosterol secreted from endophytic strain of Gymnema sylvestre for inhibition of α-glucosidase activity.

Endophytic fungi produce various types of chemicals for establishment of niche within the host plant. Due to symbiotic association, they secrete pharmaceutically important bioactive compounds and enzyme inhibitors. In this research article, we have explored the potent α-glucosidse inhibitor (AGI) produced from Fusarium equiseti recovered from the leaf of Gymnema sylvestre through bioassay-guided fraction. This study investigated the biodiversity, phylogeny, antioxidant activity and α-glucosidse inhibition of endophytic fungi isolated from Gymnema sylvestre. A total of 32 isolates obtained were grouped into 16 genera, according to their morphology of colony and spores. A high biodiversity of endophytic fungi were observed in G. sylvestre with diversity indices. Endophytic fungal strain Fusarium equiseti was identified through DNA sequencing and the sequence was deposited in GenBank database (https://ncbi.nim.nih.gov) with acession number: MF403109. The characterization of potent compound was done by FTIR, LC-ESI-MS and NMR spectroscopic analysis with IUPAC name 17-(5-ethyl-6-methylheptan-2-yl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a] phenanthren-3-ol. The isolated bioactive compound showed significant α-amylase and α-glucosidase inhibition activity with IC50 values, 4.22 ± 0.0005 µg/mL and 69.72 ± 0.001 µg/mL while IC50 values of acarbose was 5.75 ± 0.007 and 55.29 ± 0.0005 µg/mL respectively. This result is higher in comparison to other previous study. The enzyme kinetics study revealed that bioactive compound was competitive inhibitor for α-amylase and α-glucosidase. In-silico study showed that bioactive compound binds to the binding site of α-amylase, similar to that of acarbose but with higher affinity. The study highlights the importance of endophytic fungi as an alternative source of AGI (α-glucosidase inhibition) to control the diabetic condition in vitro.

Lack of induced mutagenesis in E. coli or human lymphoblast cell line upon long-term sub-culturing in medium from irradiated meat.

PURPOSE: Current study was aimed to enhance the confidence of consumers as well as entrepreneurs towards food irradiation program. MATERIALS AND METHODS: In this work, safety of high dose (25 kGy) irradiated meat samples (HDIMS) was ascertained by scoring mutation frequency through a long-term sub-culturing study in Escherichia coli MG1655 cells (ATCC 700926) up to 1500 generations (at 1%), 250 generations (at 5% and 10%) and human lymphoblast thymidine kinase heterozygote (TK6) cell line (ATCC CRL-8015) [at two gene loci, tk-/+ (thymidine kinase) and hprt+ (Hypoxanthine Phosphoribosyltransferase)] up to 156 generations using goat meat sample. Also these samples were assayed at further radiation doses of 10, 45 and 70 kGy at 2% concentration (in cell line), and 1% (in E. coli). Study was also performed with other meat samples such as chicken, fishes (pomfret and rohu) and shrimps by carrying out limited long-term sub-culturing trials in human lymphoblast cell line. Mutation analysis was also carried out using a novel DPAR (Differential loss of Plasmid Antibiotic Resistance) assay followed by sequencing of tcR (tetracycline resistance) gene of pBR322 plasmid isolated from E. coli cells grown for 1500 generations on HDIMS medium and RAPD (Random Amplified Polymorphic DNA) analysis of the genome. RESULTS AND CONCLUSION: None of the assays exhibited any induced mutation when analyzed at regular time intervals. RAPD analysis also did not indicate any change in its nucleotide sequence, ruling out the occurrence of any silent mutation. Thus, the present findings report absence of mutagenic effect of high dose irradiated meat samples.

Natural Predominance of Abscisic Acid in Pongammia pinnata ("Karanj") Honey Contributed to its Strong Antimutagenicity.

Various samples of raw (unprocessed) floral honey collected from different geographical locations of India were assayed for its antimutagenicity against ethyl methanesulfonate in E. coli MG1655 cells through rifampicin resistance assay. A monofloral honey ("Pongammia pinnata", local name "Karanj") displayed maximum antimutagenicity (78.0 ± 1.7; P ≤ 0.05). Solid phase extraction (using Amberlite XAD-2 resin) followed by HPLC resulted into different peaks displaying varying antimutagenicity. Peak at retention time (Rt) 27.9 min (henceforth called P28) displayed maximum antimutagenicity and was further characterized to be abscisic acid (ABA) using ESI-MS and NMR. Its antimutagenicity was reconfirmed through human lymphoblast cell line (TK6) mutation assay using thymidine kinase (tk+/-) cell line. Although ABA from this honey displayed strong antimutagenicity, it lacked any in vitro antioxidant capacity indicating noninvolvement of any radical scavenging in the observed antimutagenicity.