Unraveling Compositional Study, Chemometric Analysis, and Cell-Based Antioxidant Potential of Selective High Nutraceutical Value Amaranth Cultivars Using a GC-MS and NMR-Based Metabolomics Approach.

Amaranthus (family Amaranthaceae) is a potentially nutritious pseudocereal also known as a functional food owing to its high nutritional quality grains especially rich in essential amino acids. Emerging study, however, unambiguously indicates that apart from essential nutrients like protein, other phytochemicals present in amaranth seeds provide excellent health benefits. Squalene is one such phytonutrient found in Amaranthus seeds, which is also its largest vegetal source. In this research work, GC-MS and NMR spectroscopy-based metabolomics have been utilized for the compositional analysis of Amaranthus seeds coupled with a multivariate data set. Investigation of nonpolar and polar seed extracts of six different cultivars of amaranth identified 47 primary and secondary metabolites. One-way ANOVA showed significant quantitative metabolic variations in different cultivars of amaranth. Multivariate principal component analysis of both the GC-MS and NMR analyzed data broadly classified in two groups showed significant variations in the polar (lysine, arginine, GABA, and myoinositol) and nonpolar (squalene, tryptophan, and alkylated phenols, which are potential nutraceutical agents) metabolites. The squalene content estimated using HPLC varied significantly (1.61 to 4.72 mg g-1 seed dry weight) among six different cultivars. Positive correlations were found among the cellular antioxidant activity and squalene content. Cultivar AM-3 having the maximum squalene content showed the highest antioxidant activity evaluated on the cellular level over human embryonic kidney cells, clearly revealing potent intercellular reactive oxygen species scavenging capacity and strong membrane lipid peroxidation inhibition potential. Oxidative stress markers such as MDA, SOD, GSH, and CAT levels in cells further corroborated the research work. The study also indicated high concentrations of lysine (80.49 mg g-1 dry seeds) in AM-2, squalene (0.47% by weight) in AM-3, and 2,4-di-tert-butyl phenol (18.64% peak area) and myoinositol (79.07 mg g-1 dry seeds) in AM-5. This novel comparative metabolomic study successfully profiles the nutrient composition of amaranth cultivars and provides the opportunity for the development of nutraceuticals and natural antioxidants from this functional food.

Protein-polysaccharide nanoconjugates: Potential tools for delivery of plant-derived nutraceuticals.

Protein-polysaccharide nanoconjugates are covalently interactive networks that are currently the subject of intense research owing to their emerging applications in the food nanotechnology field. Due to their biocompatibility and biodegradability properties, they have played a significant role as wall materials for the formation of various nanostructures to encapsulate nutraceuticals. The food-grade protein-polysaccharide nanoconjugates would be employed to enhance the delivery and stability of nutraceuticals for their real use in the food industry. The most common edible polysaccharides (cellulose, chitosan, pectin, starch, carrageenan, fucoidan, mannan, glucomannan, and arabic gum) and proteins (silk fibroin, collagen, gelatin, soy protein, corn zein, and wheat gluten) have been used as potential building blocks in nano-encapsulation systems because of their excellent physicochemical properties. This article broadens the discussion of food-grade proteins and polysaccharides as nano-encapsulation biomaterials and their fabrication methods, along with a review of the applications of protein-polysaccharide nanoconjugates in the delivery of plant-derived nutraceuticals.

Bacterial glycobiotechnology: A biosynthetic route for the production of biopharmaceutical glycans.

The recent advancement in the human glycome and progress in the development of an inclusive network of glycosylation pathways allow the incorporation of suitable machinery for protein modification in non-natural hosts and explore novel opportunities for constructing next-generation tailored glycans and glycoconjugates. Fortunately, the emerging field of bacterial metabolic engineering has enabled the production of tailored biopolymers by harnessing living microbial factories (prokaryotes) as whole-cell biocatalysts. Microbial catalysts offer sophisticated means to develop a variety of valuable polysaccharides in bulk quantities for practical clinical applications. Glycans production through this technique is highly efficient and cost-effective, as it does not involve expensive initial materials. Metabolic glycoengineering primarily focuses on utilizing small metabolite molecules to alter biosynthetic pathways, optimization of cellular processes for glycan and glycoconjugate production, characteristic to a specific organism to produce interest tailored glycans in microbes, using preferably cheap and simple substrate. However, metabolic engineering faces one of the unique challenges, such as the need for an enzyme to catalyze desired substrate conversion when natural native substrates are already present. So, in metabolic engineering, such challenges are evaluated, and different strategies have been developed to overcome them. The generation of glycans and glycoconjugates via metabolic intermediate pathways can still be supported by glycol modeling achieved through metabolic engineering. It is evident that modern glycans engineering requires adoption of improved strain engineering strategies for creating competent glycoprotein expression platforms in bacterial hosts, in the future. These strategies include logically designing and introducing orthogonal glycosylation pathways, identifying metabolic engineering targets at the genome level, and strategically improving pathway performance (for example, through genetic modification of pathway enzymes). Here, we highlight current strategies, applications, and recent progress in metabolic engineering for producing high-value tailored glycans and their applications in biotherapeutics and diagnostics.

A global perspective on a new paradigm shift in bio-based meat alternatives for healthy diet.

A meat analogue is a casserole in which the primary ingredient is something other than meat. It goes by various other names, such as meat substitute, fake meat, alternative meat, and imitation meat. Consumers growing interest in improving their diets and the future of the planet have contributed to the move towards meat substitutes. This change is due to the growing popularity of low-fat and low-calorie diets, the rise of flexitarians, the spread of animal diseases, the loss of natural resources, and the need to cut down on carbon emissions, which lead to greenhouse effects. Plant-based meat, cultured meat, algal protein-based meat, and insect-based meat substitutes are available on the market with qualities like appearance and flavor similar to those of traditional meat. Novel ingredients like mycoprotein and soybean leg haemoglobin are mixed in with the more traditional soy proteins, cereals, green peas, etc. Plant-based meat is currently more popular in the West, but the growing interest in this product in Asian markets indicates the industry in this region will expand rapidly in the near future. Future growth in the food sector can be anticipated from technologies like lab-grown meat and its equivalents that do not require livestock breeding. Insect-based products also hold great potential as a new source of protein for human consumption. However, product safety and quality should be considered along with other factors such as marketability and affordability.

Biobased natural deep eutectic system as versatile solvents: Structure, interaction and advanced applications.

The increasing emphasis on the development of green replacements to traditional organic solvents and ionic liquids (ILs) can be attributed to the rising concerns over human health and detrimental impacts of conventional solvents towards the environment. A new generation of solvents inspired by nature and extracted from plant bioresources has evolved over the last few years, and are referred to as natural deep eutectic solvents (NADES). NADES are mixtures of natural constituents like sugars, polyalcohols, sugar-based alcohols, amino acids and organic acids. Interest in NADES has exponentially grown over the last eight years, which is evident from an upsurge in the number of research projects undertaken. NADES are highly biocompatible as they can be biosynthesized and metabolized by nearly all living organisms. These solvents pose several noteworthy advantages, such as easy synthesis, tuneable physico-chemical properties, low toxicity, high biodegradability, solute sustainability and stabilization and low melting point. Research on the applicability of NADES in diverse areas is gaining momentum, which includes as - media for chemical and enzymatic reactions; extraction media for essential oils; anti-inflammatory and antimicrobial agent; extraction of bioactive composites; as chromatographic media; preservatives for labile compounds and in drug synthesis. This review gives a complete overview of the properties, biodegradability and toxicity of NADES which we propose can assist in further knowledge generation on their significance in biological systems and usage in green and sustainable chemistry. Information on applications of NADES in biomedical, therapeutic and pharma-biotechnology fields is also highlighted in the current article along with the recent progress and future perspectives in novel applications of NADES.

Tetrahydrocannabinols: potential cannabimimetic agents for cancer therapy.

Tetrahydrocannabinols (THCs) antagonize the CB1 and CB2 cannabinoid receptors, whose signaling to the endocannabinoid system is essential for controlling cell survival and proliferation as well as psychoactive effects. Most tumor cells express a much higher level of CB1 and CB2; THCs have been investigated as potential cancer therapeutic due to their cannabimimetic properties. To date, THCs have been prescribed as palliative medicine to cancer patients but not as an anticancer modality. Growing evidence of preclinical research demonstrates that THCs reduce tumor progression by stimulating apoptosis and autophagy and inhibiting two significant hallmarks of cancer pathogenesis: metastasis and angiogenesis. However, the degree of their anticancer effects depends on the origin of the tumor site, the expression of cannabinoid receptors on tumor cells, and the dosages and types of THC. This review summarizes the current state of knowledge on the molecular processes that THCs target for their anticancer effects. It also emphasizes the substantial knowledge gaps that should be of concern in future studies. We also discuss the therapeutic effects of THCs and the problems that will need to be addressed in the future. Clarifying unanswered queries is a prerequisite to translating the THCs into an effective anticancer regime.

Microbe-fabricated nanoparticles as potent biomaterials for efficient food preservation.

In recent years, cutting-edge nanotechnology research has revolutionized several facets of the food business, including food processing, packaging, transportation, preservation, and functioning. Nanotechnology has beginning to loom large in the food business as the industry's demand for biogenic nanomaterial grows. The intracellular and extracellular synthesis of metal, metal oxide, and other essential NPs has recently been explored in a variety of microorganisms, including bacteria, actinomycetes, fungi, yeasts, microalgae, and viruses. These microbes produce a variety extracellular material, exopolysaccharides, enzymes, and secondary metabolites which play key roles in synthesizing as well as stabilizing the nanoparticle (NPs). Furthermore, genetic engineering techniques can help them to improve their capacity to generate NPs more efficiently. As a result, using microorganisms to manufacture NPs is unique and has a promising future. Microbial-mediated synthesis of NPs has lately been popular as a more environmentally friendly alternative to physical and chemical methods of nanomaterial synthesis, which require higher prices, more energy consumption, and more complex reaction conditions, as well as a potentially dangerous environmental impact. It is critical to consider regulatory measures implemented at all stages of the process, from production through refining, packaging, preservation, and storage, when producing bionanomaterials derived from culturable microbes for efficient food preservation. The current review discusses the synthesis, mechanism of action, and possible food preservation uses of microbial mediated NPs, which can assist to minimize food deterioration from the inside out while also ensuring that food is safe and free of contaminants. Despite the numerous benefits, there are looming debates concerning their usage in food items, particularly regarding its aggregation in human bodies and other risks to the environment. Other applications and impacts of these microbe-fabricated NPs in the context of future food preservation prospects connected with regulatory problems and potential hazards are highlighted.

Superior inhibition of virulence and biofilm formation of Pseudomonas aeruginosa PAO1 by phyto-synthesized silver nanoparticles through anti-quorum sensing activity.

Quorum sensing (QS)-regulated bacterial biofilm formation is a crucial issue in causing resistance against existing antibiotics. There is a considerable necessity to disrupt the interrelationship between bacterial QS, virulence, and biofilm formation. Disabling QS could be a novel tactic of great clinical importance. Here, we biosynthesized silver nanoparticles (Ka-AgNPs) using the aqueous leaf extract of Koelreuteria paniculata as a reducing and capping agents. The UV-Vis spectroscopy confirmed the synthesis of Ka-AgNPs as a characterization peak observed at 420 nm. TEM image revealed the spherical shape distribution of Ka-AgNPs with average particle size of 30.0 ± 5 nm. The anti-QS activity of Ka-AgNPs was tested against a bio-indicator bacterium Chromobacterium violaceum 12472 and a multi-drug resistant model strain of Pseudomonas aeruginosa (PAO1). The results demonstrated that the Ka-AgNPs superiorly inhibited QS-regulated virulence factors in PAO1 without affecting cell viability compared to chemically synthesized AgNPs (Cs-AgNPs). The Ka-AgNPs effectively suppressed the formation of biofilm of PAO1. RT-PCR results revealed that the Ka-AgNPs inhibited the expression of QS-regulated virulence genes of PAO1. These results suggest that the phyto-synthesized AgNPs could be used as promising anti-infective agents for treating drug-resistant P. aeruginosa.

Tailored enzymes as next-generation food-packaging tools.

Enzymes have the potential for biotransformation in the food industry. Engineering tools can be used to develop tailored enzymes for food-packaging systems that perform well and retain their activity under adverse conditions. Consequently, novel tailored enzymes have been produced to improve or include new and useful characteristics for intelligent food-packaging systems. This review discusses the protein-engineering tools applied to create new functionality in food-packaging enzymes. The challenges in applications and anticipated directions for future developments are also highlighted. The development and discovery of tailored enzymes for smart food packaging is a promising way to ensure safe and high-quality food products.

Cinnamomum verum-derived bioactives-functionalized gold nanoparticles for prevention of obesity through gut microbiota reshaping.

Existing drugs have limited success in managing obesity in human due to their low efficacy and severe side-effects. Surface-modified gold nanoparticles have now received considerable attention of researchers for efficient biomedical applications owing to their superior uptake by cells, biocompatibility, hydrophilicity and non-immunogenicity. Here we prepared Cinnamomum verum derived bioactives-functionalized gold nanoparticles (Au@P-NPs) and assessed their impact on obesity and related immune-metabolic complications in high-fat diet (HFD)-induced obese mice using metabolic experiments along with 16S RNA gene-based gut microbial profiling and faecal microbiota transplantation (FMT). Au@P-NPs treatment prevented weight gain, decreased fat deposition, reduced metabolic inflammation and endotoxaemia in HFD-fed mice. Au@P-NPs-treated group exhibited better glucose tolerance and insulin sensitivity than HFD-fed control mice, and got completely protected against hepatic steatosis. These impacts were related to increased energy expenditure and enhanced Ucp1 expression in the brown adipose tissues of Au@P-NPs-administered animals, which strongly linked with the mRNA expression of the membrane bile acid receptor TGR5. Treatment of HFD-fed animals with Au@P-NPs altered plasma bile acid profile, and increased Akkermansia muciniphila and decreased Lactobacillus populations in the faeces. Au@P-NPs-treated animals revealed altered plasma bile acid profile, and increased Akkermansia muciniphila and decreased Lactobacillus populations in the faeces. FMT experiments showed lesser weight gain and greater energy expenditure in the mice fed with faecal suspension from Au@P-NPs-treated animals than that from HFD-fed mice. These results clearly establish that gold nanoparticles functionalized with bioactive compounds of C. verum have high potential to be an anti-obesity drug.

Nanocurcumin Potently Inhibits SARS-CoV-2 Spike Protein-Induced Cytokine Storm by Deactivation of MAPK/NF-κB Signaling in Epithelial Cells.

Interleukin-mediated deep cytokine storm, an aggressive inflammatory response to SARS-CoV-2 virus infection in COVID-19 patients, is correlated directly with lung injury, multi-organ failure, and poor prognosis of severe COVID-19 patients. Curcumin (CUR), a phenolic antioxidant compound obtained from turmeric (Curcuma longa L.), is well-known for its strong anti-inflammatory activity. However, its in vivo efficacy is constrained due to poor bioavailability. Herein, we report that CUR-encapsulated polysaccharide nanoparticles (CUR-PS-NPs) potently inhibit the release of cytokines, chemokines, and growth factors associated with damage of SARS-CoV-2 spike protein (CoV2-SP)-stimulated liver Huh7.5 and lung A549 epithelial cells. Treatment with CUR-PS-NPs effectively attenuated the interaction of ACE2 and CoV2-SP. The effects of CUR-PS-NPs were linked to reduced NF-κB/MAPK signaling which in turn decreased CoV2-SP-mediated phosphorylation of p38 MAPK, p42/44 MAPK, and p65/NF-κB as well as nuclear p65/NF-κB expression. The findings of the study strongly indicate that organic NPs of CUR can be used to control hyper-inflammatory responses and prevent lung and liver injuries associated with CoV2-SP-mediated cytokine storm.

Microbial polysaccharides: An emerging family of natural biomaterials for cancer therapy and diagnostics.

Microbial polysaccharides (MPs) offer immense diversity in structural and functional properties. They are extensively used in advance biomedical science owing to their superior biodegradability, hemocompatibility, and capability to imitate the natural extracellular matrix microenvironment. Ease in tailoring, inherent bio-activity, distinct mucoadhesiveness, ability to absorb hydrophobic drugs, and plentiful availability of MPs make them prolific green biomaterials to overcome the significant constraints of cancer chemotherapeutics. Many studies have demonstrated their application to obstruct tumor development and extend survival through immune activation, apoptosis induction, and cell cycle arrest by MPs. Synoptic investigations of MPs are compulsory to decode applied basics in recent inclinations towards cancer regimens. The current review focuses on the anticancer properties of commercially available and newly explored MPs, and outlines their direct and indirect mode of action. The review also highlights cutting-edge MPs-based drug delivery systems to augment the specificity and efficiency of available chemotherapeutics, as well as their emerging role in theranostics.

Screening of cryptogamic secondary metabolites as putative inhibitors of SARS-CoV-2 main protease and ribosomal binding domain of spike glycoprotein by molecular docking and molecular dynamics approaches.

The unprecedented quick spreading of newly emerged SARS-CoV-2, the virus responsible for causing COVID-19 has put the whole world in vast crisis. Several prophylactic interventions are being performed to discover the effective anti-COVID-19 agent. Thus, the present study aims to identify the cryptogamic secondary metabolites (CSMs) as potent inhibitors of two major targets of SARS-Cov2, namely 3-chymotrypsin-like protease (3CLpro) and receptor-binding domain (RBD) of spike glycoprotein (SGP), by implementing a computational approach. Molecular docking was carried out on Autodock 4.2 software with the 3CLpro (PDB ID:6LU7) and RBD of SGP (PDB ID:6W41) of the virus. Lopinavir and Arbidol were taken as positive controls to compare the efficacy of randomly selected 53 CSMs. The drug-likeness and pharmacokinetics properties of all metabolites were accessed to discern the anti-COVID 19 activity acting well at the physiological conditions. The docking results predicted that Marchantin E and Zeorin would potentially block the catalytic site of 3CLpro with the interaction energy values of -8.42 kcal/mol and -9.04 kcal/mol, respectively. In addition, Usnic acid revealed its ability to combat the interaction of RBD of SGP to angiotensin-converting enzyme-2 in docking analysis. To certify the potent metabolites for both targets of SARS-CoV-2, MD analysis was performed for 100 ns. The results confirmed that Marchantin E could inhibit SARS-CoV-2 3CLpro and RBD of SGP as well as reveals excellent pharmacokinetic properties. The present study suggests that the identified CSMs could be quickly positioned for further experimental validation to propose promising inhibitors of SARS-CoV-2.

Product Development, Manufacturing, and Packaging of Solid Dosage Forms Under QbD and PAT Paradigm: DOE Case Studies for Industrial Applications.

An integrated approach based on QbD and PAT provides a systematic and innovative framework for product development, manufacturing, and quality risk management. In this context, the significance of the outcome of design of experiments (DOEs) to the selection of the product design, robust commercial manufacturing process, design space, and overall control strategy remains vital for the success of a drug product throughout its life cycle. This paper aims at discussing selected recent DOE case studies conducted during QbD-based and integrated QbD/PAT-based development of solid oral formulations and process improvement studies. The main focus of this paper is to highlight the rationales and importance of design selection during development and applications of mathematical models and statistical tools in analyzing DOE and PAT data for developing a design space, control strategy, and improved process monitoring. A total of 25 case studies (includes 9 PAT application studies) have been discussed in this paper which cover 11 manufacturing processes commonly utilized for solid dosage forms. Two case studies relevant to selection of packaging design for solid dosage forms are also briefly discussed to complete the scope. Overall, for a successful modern QbD approach, it is highly important that DOEs are conducted and analyzed in a logical sequence which involves designs that are phase-appropriate and quality-driven and facilitate both statistical and chemometric thinking at each development stage. This approach can result into higher regulatory flexibility along with lower economic burden during life cycle of a product, irrespective of regulatory path used (NDA or ANDA).

Phyllanthus emblica fruit extract stabilized biogenic silver nanoparticles as a growth promoter of wheat varieties by reducing ROS toxicity.

The present study is focused on the biogenic synthesis of AgNPs (B-AgNPs) using fruit extract of Phyllanthus emblica L. and its effect (0, 5, 10, 25, 50 mg/L concentrations) on early seedling growth of two wheat varieties (HD-2967 and DBW-17). The prepared silver nanoparticles were characterized with several techniques such as UV-Vis spectroscopy, powder X-ray diffraction as well as high-resolution transmission electron microscopy. The capping of AgNPs by phytochemicals was confirmed by Fourier transforms infrared (FT-IR) spectroscopy. B-AgNPs, chemically synthesized AgNPs, chemically synthesized AgNPs+10% fruit extract and AgNO3 salt were compared for phytotoxicity, based on growth parameters, ROS production, cytotoxicity assay and silver accumulation in two wheat varieties (HD-2967 and DBW-17). These effects were more pronounced in the variety HD-2967 over DBW-17 variety at 10 mg/L B-AgNPs exposure. Root cells viability of treated radicles was studied using Evans blue dye assay which suggest that 10 mg/L B-AgNPs was effective in promoting early seedling growth by decreasing ROS toxicity. Lower accumulation of Ag resulting in higher root cell viability than those of chemically synthesized AgNPs treated seedlings. The findings of the present study clearly indicate that phytochemicals capped AgNPs act as a growth promoter at lower concentrations by delivering a potent antioxidant during early seedling growth as compared to chemically synthesized AgNPs treated wheat seedlings.

ZnO/Curcumin Nanocomposites for Enhanced Inhibition of Pseudomonas aeruginosa Virulence via LasR-RhlR Quorum Sensing Systems.

The indiscriminate and excessive use of antibiotics has ultimately led to the emergence of bacterial resistant mutants or superbugs. These superbugs are difficult to control with conventional antibiotics. Disabling quorum sensing (QS), a population-density-dependent cell-to-cell communication process used by bacteria to coordinate the expression of virulence genes and biofilm formation, with dietary phytochemicals is emerging as a non-antibiotic strategy to inhibit bacterial pathogenicity. Although curcumin is an anti-QS agent and its delivery to cells has been a challenge due to poor bioavailability, ZnO/curcumin nanocomposites (ZnC-NCs) were fabricated with enhanced delivery of curcumin inside the bacterial superbug Pseudomonas aeruginosa PAO1 for effective inhibition of its QS and biofilm formation. Sustained release of curcumin from ZnC-NCs was observed where 51% curcumin at pH 7.2 and 83% curcumin at pH 5.5 were released within 48 h. ZnC-NCs also decreased the production of virulence factors and biofilm formation without affecting planktonic cell growth. Both LasR and RhlR QS systems were inhibited by ZnC-NCs. ZnC-NCs were also capable of protecting both mice as well as lung epithelial cells from killing by PAO1. The superoxide anions (O2·-) were also found as key players in suppressing PAO1 QS systems by ZnC-NCs. Overall, ZnC-NCs enhanced curcumin bioavailability for effective inhibition of QS signaling in P. aeruginosa via LasR-RhlR suppression and O2·- generation.