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.

Release of an encrypted, highly potent ACE-inhibitory peptide by enzymatic hydrolysis of moth bean (Vigna aconitifolia) protein.

Aim: Dietary approaches for the regulation of blood pressure are the need of the hour. Hence, identifying the foods possessing such activity is gaining importance. With this aim, moth bean (Vigna aconitifolia), an underutilized pulse, was explored for the presence of antihypertensive activity in terms of angiotensin converting enzyme (ACE)-inhibition bioactivity. Methods: Defatted moth bean protein concentrate was hydrolyzed by using different proteases including Alcalase, papain, and trypsin, to identify the enzyme producing highly potent ACE inhibitory peptides. The hydrolysate showing the highest ACE inhibitory activity was further fractionated using an ultrafiltration membrane (10, 3 and 1 kDa) based on ACE inhibitory activity. The active fraction was further subjected to the ion-exchange chromatography followed by RP-HPLC and LC-MS/MS analysis for the enrichment and identification of ACE inhibitory peptides. Finally, based on the bioinformatic analysis, few peptides were synthesized and evaluated for ACE inhibitory activity, followed by docking study and molecular dynamic simulation of a peptide with the highest ACE inhibitory activity. Results and discussion: Out of the three proteases, Alcalase-derived hydrolysate showed the highest (~59%) ACE inhibition activity. Molecular weight-based fractionation revealed that <1 kDa fraction possessed the highest ACE inhibitory activity. Activity guided separation of 1 kDa fraction using ion-exchange chromatography, RP-HPLC and LC-MS/MS showed the presence of about 45 peptides. Based on the bioinformatic analysis, 15 peptides were synthesized and evaluated for ACE inhibitory activity. Among these, a novel octapeptide FPPPKVIQ showed the highest ACE inhibitory activity (93.4%) with an IC50 of 0.24 µM. This peptide retained about 59% activity post gastrointestinal digestion simulation. A Dixon plot as well as docking studies revealed the uncompetitive inhibitory nature of this peptide with a Ki value of 0.81 µM. Molecular dynamic simulation studies till 100 ns ensured the stability of the ACE-peptide complex. Conclusion: Thus, present study identified a novel potent ACE inhibitory peptide from moth bean that can be incorporated in a functional dietary formulation for regulation of hypertension.

Gamma radiation processing for extending shelf-life and ensuring quality of minimally processed ready-to-eat onions.

Onions are always in high demand owing to various culinary as well as health protective properties and these days there is increased consumer preference for ready-to-eat or ready-to-cook onions. In this context, the current study was aimed to extend the keeping quality of minimally processed onions for an extended period while ensuring microbial safety as well as sprouting inhibition through an integrated approach. The optimized combinatorial approach included gamma radiation treatment (Dmin60 Gy), minimal processing (de-skinning and scooping) and packaging in trays wrapped with polypropylene (PP; 10 µm thick) film followed by storage at low temperature (4-6 °C, relative humidity RH 65-70%). The parameters like shelf life, physico-chemical (colorimetry, moisture), organoleptic and nutritional properties were comprehensively assessed and found to be well retained up to 30 days with moisture loss of ≤ 5% and overall acceptability rating of 7 on 9-point hedonic scale. Microbiological analyses confirmed absence of Salmonella spp in these stored onions thus ensuring microbial safety. Nutritional profiling including carbohydrate, protein, fat, energy, and ash content revealed no significant change due to the processing as well as during storage. Thus, the radiation processing of freshly harvested bulbs followed by minimal processing, packing in formulated package and storage under low temperature conditions were found acceptable up to 30 days in the ready-to-eat form. Current findings provide credible evidences ascertaining extended shelf-life as well ensuring microbial safety of processed onions for commercial utilization by the food industries.

'BhAVI-23'-A spice-herb based dietary infusion possessing in-vitro anti-viral potential.

BACKGROUND: Viruses cause many life threatening human diseases. Recently, COVID-19 pandemic has challenged the health care systems worldwide. As a disease preventive approach and to bring relief to the severity of the symptoms, a infusion termed as Bhabha Anti-Viral Infusion-23 ('BhAVI-23') was conceptualized and formulated which comprised of 23 selected spices and herbals. OBJECTIVE: The present study was conducted to assess the in vitro antiviral potential of the formulation, BhaAVI-23. MATERIAL AND METHODS: The in-vitro anti-viral potential of BhAVI-23 was assessed through inhibition of HIV1 reverse transcriptase (RT) as well as through a novel P1 (virulent) bacteriphage based screening assay system. Anti-diabetic potential was assessed by non-enzymatic glycosylation of haemoglobin and the bioactive volatile components were detected through headspace gas chromatography followed by molecular docking analysis. RESULTS: The infusion displayed prominent anti-viral activity as evident from significant (57%) inhibition of the HIV1-RT as well as through reduction in the infectivity of P1 (virulent) bacteriophage. The infusion also exerted profound protection (∼64%) to non-enzymatic glycosylation of haemoglobin. Headspace gas chromatography and mass spectrometric analysis confirmed the presence of at least 47 major compounds. Docking analysis indicated possible interaction of α-pinene and eugenol with SARS-CoV spike protein. CONCLUSION: This 'BhAVI-23' infusion displayed prominent in-vitro anti-viral and anti-diabetic potential in different model systems. These attributes have relevance as diabetic patients are more prone to COVID-19 morbidity. 'BhAVI-23' opens the avenue for its potential inclusion as a supportive health care system upon due regulatory approval during the current pandemic.

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.

Formulation of a nasogastric liquid feed and shelf-life extension using gamma radiation.

Nasogastric liquid feed formulation (NGLF) was developed for immunocompromised patients who are vulnerable targets of pathogenic assault. NGLF consisted of cereals, pulses, vegetables, and milk powder to provide balanced nutrients; however, the shelf life was only a few hours because this product was highly prone to microbial contamination and proliferation due to its high water content and rich nutrients. Postpreparation storage and distribution was very difficult, even at chilled temperatures. To overcome this problem, the NGLF was irradiated at various doses (2.5 to 10 kGy). Gamma irradiation at 10 kGy reduced the microbial load to nondetectable levels, and the product could be stored up to 1 month without any detectable increase in microbial load. The sensory evaluation did not indicate differences between the nonirradiated fresh, irradiated fresh, and stored samples. Nutritional quality in terms of total carbohydrates, dietary fiber, proteins, calories, vitamins A and C, and the micronutrients calcium, iron, and zinc was not affected by irradiation. NGLF also possessed antimutagenic potential against ethylmethanesulphonate-induced mutagenesis in Escherichia coli cells as evaluated by the rifampin resistance assay. This property of NGLF remained unchanged even after exposure to a 10-kGy dose of gamma radiation. Thus, irradiated NGLF seemed to be a safe and wholesome food for immunocompromised patients.

Induction of apoptosis in human cancer cells by a Bacillus lipopeptide bacillomycin D.

A newly isolated and characterized Bacillus amyloliquefaciens strain fiply 3A has been found to produce an extracellular cyclic lipopeptide which structurally resembled bacillomycin D, earlier reported to be produced by Bacillus subtilis. The lipopeptide showed a dose dependent killing of three different human cancer cell lines viz. A549 (alveolar adenocarcinoma), A498 (renal carcinoma) and HCT-15 (colon adenocarcinoma), while not affecting the normal cell line L-132 (pulmonary epithelial cells) when analyzed using MTT assay and FACS analysis. Staining the cells with H2-DCFDA showed an increase in reactive oxygen species (ROS) formation in the lipopeptide treated cell population. Hoechst 33342 staining of nuclei further indicated apoptosis as a major mechanism of cell death in lipopeptide treated cells and the typical symptoms of apoptosis including cell shrinkage, nuclear condensation and fragmentation of nuclei were observed. Lipopeptide treatment induced extensive DNA damage in the treated cells, which was indicated by a TUNEL assay. Flow cytometric analysis exhibited lipopeptide concentration dependent apoptosis which was further confirmed during clonogenic assay of the lipopeptide treated cells.

Effectiveness of radiation processing for elimination of Salmonella Typhimurium from minimally processed pineapple (Ananas comosus Merr.).

The microbiological quality of market samples of minimally processed (MP) pineapple was examined. The effectiveness of radiation treatment in eliminating Salmonella Typhimurium from laboratory inoculated ready-to-eat pineapple slices was also studied. Microbiological quality of minimally processed pineapple samples from Mumbai market was poor; 8.8% of the samples were positive for Salmonella. D(10) (the radiation dose required to reduce bacterial population by 90%) value for S. Typhimurium inoculated in pineapple was 0.242 kGy. Inoculated pack studies in minimally processed pineapple showed that the treatment with a 2-kGy dose of gamma radiation could eliminate 5 log CFU/g of S. Typhimurium. The pathogen was not detected from radiation-processed samples up to 12 d during storage at 4 and 10 degrees C. The processing of market samples with 1 and 2 kGy was effective in improving the microbiological quality of these products.