Untapped potential of non-conventional rubus species: bioactivity, nutrition, and livelihood opportunities.

Rubus species holds promise as a valuable source of polyphenols and bioactive compounds, offering significant potential as functional food ingredients with both nutraceutical and pharmaceutical benefits. However, many edible species within this genus remain under-explored and their importance is largely unrecognized. This review aims to provide an overview of the nutritional and bioactive components of both explored and under-explored Rubus species, highlighting their potential health advantages, value addition, and recent advancements. The economic exploitation of Rubus is currently limited to a few cultivated species, while numerous non-conventional and wild edible species are overlooked. Recognizing the economic and nutritional significance of exploited Rubus species, it is imperative to explore the untapped potential of these underutilized plants. By doing so, these species can be preserved from endangerment and contribute to nutritional and livelihood security for communities having access to them. This review emphasizes the importance of understanding the exceptional characteristics of Rubus species as "superfoods" and encourages the promotion and cultivation of these unexplored species. By expanding the cultivation and utilization of under-explored Rubus species, we can unlock their full potential and support sustainable nutritional and economic benefits.

Nutritional advantages of barnyard millet and opportunities for its processing as value-added foods.

Barnyard millet (Echinochloa species) has received appreciable attention for its susceptibility to biotic and abiotic stresses, multiple harvests in a year and rich in micronutrients, fibers and phytochemicals. It is believed that the consumption of barnyard millet can possess various health benefits against diabetes, cardiovascular diseases, obesity, skin problems, cancer and celiac disease. The flour of barnyard millet is gluten-free and can be incorporated into the diet of celiac and diabetic patients. Considering the nutritional value of millet, various millet-based food products like bread, snack, baby foods, millet wine, porridge, fast foods and millet nutrition powder can be prepared. Future research and developments on barnyard millet and its products may help cope with various diseases known to humans. This paper discusses barnyard millet's nutritional and health benefits as whole grain and its value-added products. The paper also provides insights into the processing of barnyard millet and its effect on the functional properties and, future uses of barnyard millet in the field food industry as ready-to-cook and ready-to-eat products as well as in industrial uses, acting as a potential future crop contributing to food and nutritional security.

Trends in functional beverages: Functional ingredients, processing technologies, stability, health benefits, and consumer perspective.

The World Health Organization's emphasis on the health benefits of functional foods and beverages that has contributed to the rise in its popularity globally. Besides these consumers have become more aware of the importance of their food composition and nutrition. Among the fastest-growing market segments within the functional food industries, the functional drinks market focuses on fortified beverages or products that are novel with improved bioavailability of bioactive compounds, and their implicated health benefits. The bioactive ingredients in functional beverages include phenolic compounds, minerals, vitamins, amino acids, peptides, unsaturated fatty acids, etc. which can be obtained from plant, animal and microorganisms. The types of functional beverages which are globally intensifying the markets are pre-/pro-biotics, beauty drinks, cognitive and immune system enhancers, energy and sports drink produced via several thermal and non-thermal processes. Researchers are focusing on improving the stability of the active compounds by encapsulation, emulsion, and high-pressure homogenization techniques to strengthen the positive consumer perspective in functional beverages. However, more research is needed in terms of bioavailability, consumer safety, and sustainability of the process. Hence, product development, storage stability, and sensory properties of these products are vital for consumer acceptance. This review focuses on the recent trends and developments in the functional beverages industry. The review provides a critical discussion on diverse functional ingredients, bioactive sources, production processes, emerging process technologies, improvement in the stability of ingredients and bioactive compounds. This review also outlines the global market and consumer perception of functional beverages with the future perspective and scope.

Trends in millet and pseudomillet proteins - Characterization, processing and food applications.

Millets are small-seeded crops which have been well adopted globally owing to their high concentration of macro and micronutrients such as protein, dietary fibre, essential fatty acids, minerals and vitamins. Considering their climate resilience and potential role in nutritional and health security, the year 2023 has been declared as 'International Year of Millets' by the United Nations. Cereals being the major nutrient vehicle for a majority population, and proteins being the second most abundant nutrient in millets, these grains can be a suitable alternative for plant-based proteins. Therefore, this review was written with an aim to succinctly provide an overview of the available literature take on the characterization, processing and applications of millet-based proteins. This information would play an important role in realizing the research gap restricting the utilization of complete potential of millet proteins. This can be further used by researchers and food industries for understanding the scope of millet proteins as an ingredient for novel food product development.

Beetroot as a novel ingredient for its versatile food applications.

Beta vulgaris, also known as Beetroot, is a member of a family of Chenopodiaceae and is widely used as a natural food colorant. It gets its distinctive color due to nitrogen-containing water-soluble pigments betalains. Beetroot is an exquisite cradle of nutrients, including proteins, sucrose, carbohydrates, vitamins (B complex and vitamin C), minerals, fiber. They also contain an appreciable amount of phenolic compounds and antioxidants such as coumarins, carotenoids, sesquiterpenoids, triterpenes, flavonoids (astragalin, tiliroside, rhamnocitrin, kaempferol, rhamnetin). Recent studies evidenced that beetroot consumption had favorable physiological benefits, leading to improved cardiovascular diseases, hypertension, diabetes, cancer, hepatic steatosis, liver damage, etc. This review gives insights into developing beetroot as a potential and novel ingredient for versatile food applications and the latest research conducted worldwide. The phytochemical diversity of beetroot makes them potential sources of nutraceutical compounds from which functional foods can be obtained. The article aimed to comprehensively collate some of the vital information published on beetroot incurred in the agri-food sector and a comprehensive review detailing the potentiality of tapping bioactive compounds in the entire agriculture-based food sector.

Kaempferol: A flavonoid with wider biological activities and its applications.

Kaempferol and its derivatives are naturally occurring phytochemicals with promising bioactivities. This flavonol can reduce the lipid oxidation in the human body, prevent the organs and cell structure from deterioration and protect their functional integrity. This review has extensively highlighted the antioxidant, antimicrobial, anticancer, neuroprotective, and hepatoprotective activity of kaempferol. However, poor water solubility and low bioavailability of kaempferol greatly limit its applications. The utilization of advanced delivery systems can improve its stability, efficacy, and bioavailability. This is the first review that aimed to comprehensively collate some of the vital information published on biosynthesis, mechanism of action, bioactivities, bioavailability, and toxicological potential of kaempferol. Besides, it provides insights into the future direction on the improvement of bioavailability of kaempferol for wide applications.

Multi-dose Formulation Development for a Quadrivalent Human Papillomavirus Virus-Like Particle-Based Vaccine: Part II- Real-time and Accelerated Stability Studies.

This work describes Part 2 of multi-dose formulation development of a Human Papillomavirus (HPV) Virus-Like Particle (VLP) based vaccine (see Part 1 in companion paper). Storage stability studies with candidate multi-dose formulations containing individual or combinations of seven different antimicrobial preservatives (APs) were performed with quadrivalent HPV VLP (6, 11, 16, 18) antigens adsorbed to aluminum-salt adjuvant (Alhydrogel®). Real-time (up to two years, 2-8°C) and accelerated (months at 25 and 40°C) stability studies identified eight lead candidates as measured by antigen stability (competitive ELISA employing conformational serotype-specific mAbs), antimicrobial effectiveness (modified European Pharmacopeia assay), total protein content (SDS-PAGE), and AP concentration (RP-UHPLC). The AH-adsorbed HPV18 VLP component was most sensitive to AP-induced destabilization. Optimal quadrivalent antigen storage stability while maintaining antimicrobial effectiveness was observed with 2-phenoxyethanol, benzyl alcohol, chlorobutanol, and 2-phenoxyethanol + benzyl alcohol combination. Interestingly, for single-AP containing multi-dose formulations, this rank-ordering of storage stability did not correlate with previously reported biophysical measurements of AP-induced antigen destabilization. Moreover, other APs (e.g., m-cresol, phenol, parabens) described by others for inclusion in multi-dose HPV VLP formulations showed suboptimal stability. These results suggest that each HPV VLP vaccine candidate (e.g., different serotypes, expression systems, processes, adjuvants) will require customized multi-dose formulation development.

Multi-Dose Formulation Development for a Quadrivalent Human Papillomavirus Virus-Like Particle-Based Vaccine: Part I - Screening of Preservative Combinations.

The development of multi-dose, subunit vaccine formulations can be challenging since antimicrobial preservatives (APs) often destabilize protein antigens. In this work, we evaluated Human Papillomavirus (HPV) Virus-Like Particles (VLPs) to determine if combining different APs used in approved parenteral products, each at lower concentrations than used alone, would maintain both antimicrobial effectiveness and antigen stability. To identify promising AP combinations, two different screening strategies were utilized: (1) empirical one-factor-at-a-time (OFAT) and (2) statistical design-of-experiments (DOE). Seven different APs were employed to screen for two- and three-AP combinations using high-throughput methods for antimicrobial effectiveness (i.e., microbial growth inhibition assay and a modified European Pharmacopeia method) and antigen stability (i.e., serotype-specific mAb binding to conformational epitopes of HPV6, 11, 16 VLPs by ELISA). The OFAT and DOE approaches were complementary, such that initial OFAT results (and associated lessons learned) were subsequently employed to optimize the combinations using DOE. Additional validation experiments confirmed the final selection of top AP-combinations predicted by DOE modeling. Overall, 20 candidate multi-dose formulations containing two- or three-AP combinations were down-selected. As described in Part 2 (companion paper), long-term storage stability profiles of aluminum-adjuvanted, quadrivalent HPV VLP formulations containing these lead candidate AP combinations are compared to single APs.

Multivariate analysis of chemometric based aroma dynamics in small cardamom (Elettaria cardamomum Maton) during drying.

The aroma compositions and essential oil contents extracted from small cardamom (Elettaria cardamomum Maton) dried at three temperatures using a hot air convective dryer was studied by chromatographic methods. The major compounds in the dried samples were 1,8-cineole (7.23-11.76%) and α-Terpinyl acetate (61.65-68.19%) that were responsible for the specific aroma. The extracted essential oil was found to be majorly composed of alkenes (33-42%), alcohols (18-30%) and esters (18-22%) with acids, alkyls and ketones. The changes in esters and alcohols with drying time were more significant (p < 0.05) than acids, alkenes and aldehydes at all three temperatures. The data were analysed using Principal Component Analysis (PCA) and Hierarchical Cluster Analysis. The analyses indicated that the compounds varied significantly (p < 0.05) with drying temperatures. The PCA revealed that there was overall 66.47, 75.96 and 78.25% cumulative data variance among the constituents of essential oils of cardamom samples dried at 40, 50 and 60 °C respectively. The samples showed a significant (p < 0.05) reduction in aroma constituents after 26, 18 and 16 h of drying at 40, 50 and 60 °C respectively. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-022-05560-8.

Aflatoxins in Cereals and Cereal-Based Products: Occurrence, Toxicity, Impact on Human Health, and Their Detoxification and Management Strategies.

Cereals and cereal-based products are primary sources of nutrition across the world. However, contamination of these foods with aflatoxins (AFs), secondary metabolites produced by several fungal species, has raised serious concerns. AF generation in innate substrates is influenced by several parameters, including the substrate type, fungus species, moisture content, minerals, humidity, temperature, and physical injury to the kernels. Consumption of AF-contaminated cereals and cereal-based products can lead to both acute and chronic health issues related to physical and mental maturity, reproduction, and the nervous system. Therefore, the precise detection methods, detoxification, and management strategies of AFs in cereal and cereal-based products are crucial for food safety as well as consumer health. Hence, this review provides a brief overview of the occurrence, chemical characteristics, biosynthetic processes, health hazards, and detection techniques of AFs, along with a focus on detoxification and management strategies that could be implemented for food safety and security.

Analytical and Preformulation Characterization Studies of Human Papillomavirus Virus-Like Particles to Enable Quadrivalent Multi-Dose Vaccine Formulation Development.

Introducing multi-dose formulations of Human Papillomavirus (HPV) vaccines will reduce costs and enable improved global vaccine coverage, especially in low- and middle-income countries. This work describes the development of key analytical methods later utilized for HPV vaccine multi-dose formulation development. First, down-selection of physicochemical methods suitable for multi-dose formulation development of four HPV (6, 11, 16, and 18) Virus-Like Particles (VLPs) adsorbed to an aluminum adjuvant (Alhydrogel®, AH) was performed. The four monovalent AH-adsorbed HPV VLPs were then characterized using these down-selected methods. Second, stability-indicating competitive ELISA assays were developed using HPV serotype-specific neutralizing mAbs, to monitor relative antibody binding profiles of the four AH-adsorbed VLPs during storage. Third, concentration-dependent preservative-induced destabilization of HPV16 VLPs was demonstrated by addition of eight preservatives found in parenterally administered pharmaceuticals and vaccines, as measured by ELISA, dynamic light scattering, and differential scanning calorimetry. Finally, preservative stability and effectiveness in the presence of vaccine components were evaluated using a combination of RP-UHPLC, a microbial growth inhibition assay, and a modified version of the European Pharmacopoeia assay (Ph. Eur. 5.1.3). Results are discussed in terms of analytical challenges encountered to identify and develop high-throughput methods that facilitate multi-dose formulation development of aluminum-adjuvanted protein-based vaccine candidates.

Bioactive potential of beetroot (Beta vulgaris).

Beetroot (Beta vulgaris) is the most well-known and commonly cultivated fruit from the Chenopodiaceae family. Beetroot is a rich source of nutrients including vitamins (B complex and C), minerals, fibre, proteins, and a variety of bioactive phenolic substances, which are chiefly composed of betalains, and other components possessing antioxidant activity, such as coumarins, carotenoids, sesquiterpenoids, triterpenes, and flavonoids (astragalin, tiliroside, rhamnocitrin, kaempferol, rhamnetin). Beetroot and its value-added products provide a variety of health advantages and may help prevent and manage various ailments and diseases due to bioactive components. Beetroot's phytochemical diversity makes them potential sources of nutraceutical chemicals that can be used to build functional foods. Pharmacologically, beetroot has the potential to be an antioxidant, antimicrobial, anticancerous, hypocholesterolemic, and anti-inflammatory agent. In a comprehensive analysis, this review first provides an overview of the bioactive compounds present in beetroot and its parts, followed by a specific description of the current evidence on this bioactive potential of beetroot and its parts, highlighting the biochemical mechanisms involved. Additionally, the factors affecting the concentration and activity of the beetroot bioactives and the best possible method to conserve its bioactivity has also been discussed in this review.

Ultrasound-assisted modification of gelation properties of proteins: A review.

Protein gels have diverse applications in the food, pharmaceutical, and cosmetic sectors due to their affordability, biodegradability, and edibility. However, the inherent properties of some native proteins have a few drawbacks that have to be tailored to meet the needs of specific functions as the food ingredients. The protein gelation properties mainly depend on the protein molecular structure, primarily the folding and unfolding of secondary structural elements (α-helix and ß-sheets) with distinctive functions. In the past, a great amount of work (thermal, chemical, and enzymatic methods) has been carried out to enhance the gelation and functional properties of proteins. Recently, the traditional methods have been replaced with non-thermal physical methods that enhance the properties for better applications. One such approach is the use of ultrasonic technology as a low-cost green technology to modify the molecular orientation attributed to the native chemistry and functionality of that proteins. Ultrasonic technology is important in food systems and can be effectively used as an alternative method to improve the protein gelling characteristics to form high-quality gels. This article is aimed to comprehensively collate some of the vital information published on the mechanism of protein gelation by ultrasonication and review the effects of ultrasound-assisted extraction and treatments on gelation properties of different proteins. The enhanced gelation properties by the ultrasound application open a new stage of technology that enables the proteins for better utilization in the food processing sector.

Citrinin Mycotoxin Contamination in Food and Feed: Impact on Agriculture, Human Health, and Detection and Management Strategies.

Citrinin (CIT) is a mycotoxin produced by different species of Aspergillus, Penicillium, and Monascus. CIT can contaminate a wide range of foods and feeds at any time during the pre-harvest, harvest, and post-harvest stages. CIT can be usually found in beans, fruits, fruit and vegetable juices, herbs and spices, and dairy products, as well as red mold rice. CIT exerts nephrotoxic and genotoxic effects in both humans and animals, thereby raising concerns regarding the consumption of CIT-contaminated food and feed. Hence, to minimize the risk of CIT contamination in food and feed, understanding the incidence of CIT occurrence, its sources, and biosynthetic pathways could assist in the effective implementation of detection and mitigation measures. Therefore, this review aims to shed light on sources of CIT, its prevalence in food and feed, biosynthetic pathways, and genes involved, with a major focus on detection and management strategies to ensure the safety and security of food and feed.

A first-in-human phase 0 clinical study of RNA interference-based spherical nucleic acids in patients with recurrent glioblastoma.

Glioblastoma (GBM) is one of the most difficult cancers to effectively treat, in part because of the lack of precision therapies and limited therapeutic access to intracranial tumor sites due to the presence of the blood-brain and blood-tumor barriers. We have developed a precision medicine approach for GBM treatment that involves the use of brain-penetrant RNA interference-based spherical nucleic acids (SNAs), which consist of gold nanoparticle cores covalently conjugated with radially oriented and densely packed small interfering RNA (siRNA) oligonucleotides. On the basis of previous preclinical evaluation, we conducted toxicology and toxicokinetic studies in nonhuman primates and a single-arm, open-label phase 0 first-in-human trial (NCT03020017) to determine safety, pharmacokinetics, intratumoral accumulation and gene-suppressive activity of systemically administered SNAs carrying siRNA specific for the GBM oncogene Bcl2Like12 (Bcl2L12). Patients with recurrent GBM were treated with intravenous administration of siBcl2L12-SNAs (drug moniker: NU-0129), at a dose corresponding to 1/50th of the no-observed-adverse-event level, followed by tumor resection. Safety assessment revealed no grade 4 or 5 treatment-related toxicities. Inductively coupled plasma mass spectrometry, x-ray fluorescence microscopy, and silver staining of resected GBM tissue demonstrated that intravenously administered SNAs reached patient tumors, with gold enrichment observed in the tumor-associated endothelium, macrophages, and tumor cells. NU-0129 uptake into glioma cells correlated with a reduction in tumor-associated Bcl2L12 protein expression, as indicated by comparison of matched primary tumor and NU-0129-treated recurrent tumor. Our results establish SNA nanoconjugates as a potential brain-penetrant precision medicine approach for the systemic treatment of GBM.

Assessment of 3D printability of composite dairy matrix by correlating with its rheological properties.

In this study, the potential of heat desiccated milk powder (HDMP) in a composite dairy matrix with semi skimmed milk powder (SSMP) was assessed for hot melt extrusion-based 3D printing. The rheological characteristics of formulations at three stages i.e., pre-printing, printing, and post-printing were investigated. The shear thinning with rapid shear recovery and thermoresponsive behavior of the formulations were analyzed to mimic the prevailing conditions of pre-processing, processing, and post-processing of formulations to understand the temperature induced variations in their rheological characteristics during each stage. The rheological properties were correlated with printability through assessment of the consistency of straight lines (1D), average area of lattice scaffolds (2D), and dimensional stability of the 3D printed constructs. Results demonstrated that an increase in the level of incorporation of SSMP and a decrease in the proportion of HDMP increased the shear thinning behavior, viscosity (ɳ), yield stress (τ0), storage modulus (G') and a decline in the shear recoverability of the formulations. The thermoresponsive behavior of the formulations was established with gelation temperature ranging from 28.1 to 29.4˚C. The formulation SSMP (35): HDMP (25) resulted in sagging of the printed constructs, whereas the formulation SSMP (55): HDMP (5.0) exhibited the highest dimensional stability and shape retention post printing, owing to its maximum τ0 (1211.8 Pa) and G' (7026.4 Pa). The results obtained could provide insight into improving the performance of an HME based 3D printing in the dairy and food industries.

Methods for in vitro modeling of glioma invasion: Choosing tools to meet the need.

Widespread tumor cell invasion is a fundamental property of diffuse gliomas and is ultimately responsible for their poor prognosis. A greater understanding of basic mechanisms underlying glioma invasion is needed to provide insights into therapies that could potentially counteract them. While none of the currently available in vitro models can fully recapitulate the complex interactions of glioma cells within the brain tumor microenvironment, if chosen and developed appropriately, these models can provide controlled experimental settings to study molecular and cellular phenomena that are challenging or impossible to model in vivo. Therefore, selecting the most appropriate in vitro model, together with its inherent advantages and limitations, for specific hypotheses and experimental questions achieves primary significance. In this review, we describe and discuss commonly used methods for modeling and studying glioma invasion in vitro, including platforms, matrices, cell culture, and visualization techniques, so that choices for experimental approach are informed and optimal.

Inhibition ELISA as a putative tool for the identification and quantification of meningococcal A and X polysaccharides at various stages of vaccine development.

The multivalent glycoconjugate vaccines against Neisseria meningitidis are extremely high-priced for the developing world. The high cost is due to the manufacturing set-up required to produce an effective vaccine and other inflators like complex production steps including the production and purification of the polysaccharide and consequently its conjugation with a protein and finally formulating the finished multivalent product. There is an urgent need for assays which are simple, precise, can be applicable at multiple steps and contribute in reducing the overall manufacturing cost, thereby making the vaccines more equitable to the developing world. WHO recommends serological tests for polysaccharide identification and quantitation at different stages of conjugate vaccine production. We report development of inhibition ELISAs for the identification and quantification of N. meningitidis serogroup A (MenA) and N. meningitidis serogroup X (MenX) polysaccharides (PSs) in samples from stage of cell banking till production of finished product. The method was qualified on various parameters such as specificity, intermediate precision, sensitivity and accuracy. Our results provide a proof of concept for the use of an inhibition ELISA as a common tool for the identification and quantification of PS at various stages of vaccine development and manufacture.