Bioactive compounds of Aegle marmelos L., medicinal values and its food applications: A critical review.

Aegle marmelos L. (bael) is a fruit tree of Rutaceae family, widely grown all over the world. This plant is gaining popularity because of its nutrient-rich fruits and immense traditional medicinal usage and pharmacological properties. The health promotive and protective effect of bael fruit is accounted by fibers, carotenoids, phenolics, terpenoids, coumarins, flavonoids, and alkaloids. The curative relevance of these compounds has been assessed by various in vivo and in vitro studies. Fruit shows numerous possible health benefits, namely, radio-protective effects, peroxidation, antibacterial, inhibition of lipid, antidiarrheal, gastroprotective, antiviral, antidiabetic, anti-ulcerative colitis, cardioprotective, free-radical scavenging (antioxidant) and hepatoprotective effects. The health benefits of bael are not only limited to edible portion (fruit), but it also extends to nonedible portion (root, trunk, bark, leaf, flower and seed) having comparable biologically active compounds. Increasing awareness about the role of diet among health-conscious consumers for human well-being has increased the interest in functional foods thereby exploration of the functional attributes of various underutilized plants is being reaffirmed and various sources are emerged out as suitable food material for processing industry. The various scientific reports collected from different bibliometric sources suggested that A. marmelos and its bioactive constituents could play a vital role in the prevention of several chronic and degenerative diseases associated with oxidation stress. This review emphasis on recent scientific evidences on nutrition and bioactive profile of A. marmelos, health benefits along with clinical and nonclinical trials of various phytoconstituents and A. marmelos potential in food processing industry for various food products. Our study suggests that this plant does indeed have pharmacological properties of interest, however, further extensive research is needed to establish a potential strategy that can balance the pharmacological and toxic effects of bael.

Functional characterization of naturally occurring genetic variations of the human guanine-rich RNA sequence binding factor 1 (GRSF1).

The guanine-rich RNA sequence binding factor 1 (GRSF1) constitutes an ubiquitously occurring RNA-binding protein (RBP), which belongs to the family of heterogeneous nuclear ribonucleoprotein F/H (hnRNP F/H). It has been implicated in nuclear, cytosolic and mitochondrial RNA metabolism. Although the crystal structures of GRSF1 orthologs have not been solved, amino acid alignments with similar RNA-binding proteins suggested the existence of three RNA-binding domains designated quasi-RNA recognition motifs (qRRMs). Here we established 3D-models for the three qRRMs of human GRSF1 on the basis of the NMR structure of hnRNP F and identified the putative RNA interacting amino acids. Next, we explored the genetic variability of the three qRRMs of human GRSF1 by searching genomic databases and tested the functional consequences of naturally occurring mutants. For this purpose the RNA-binding capacity of wild-type and mutant recombinant GRSF1 protein species was assessed by quantitative RNA electrophoretic mobility shift assays. We found that some of the naturally occurring GRSF1 mutants exhibited a strongly reduced RNA-binding activity although the general protein structure was hardly affected. These data suggested that homozygous allele carriers of these particular mutants express dysfunctional GRSF1 and thus may show defective GRSF1 signaling.

Functional characterization of isolated RNA-binding domains of the GRSF1 protein.

The Guanine-rich RNA sequence binding factor 1 (GRSF1) is a member of the heterogeneous nuclear ribonucleoprotein F/H family and has been implicated in RNA processing, RNA transport and translational regulation. Amino acid alignments and homology modeling suggested the existence of three distinct RNA-binding domains and two auxiliary domains. Unfortunately, little is known about the molecular details of GRSF1/RNA interactions. To explore the RNA-binding mechanisms we first expressed full-length human GRSF1 and several truncation mutants, which include the three separated qRRM domains in E. coli, purified the recombinant proteins and quantified their RNA-binding affinity by RNA electrophoretic mobility shift assays. The expression levels varied between 1 and 10mg purified protein per L bacterial liquid culture and for full-length human GRSF1 a binding constant (KD-value) of 0.5µM was determined. In addition, our mechanistic experiments with different truncation mutants allowed the following conclusions: i) Deletion of either of the three RNA-binding domains impaired the RNA-binding affinity suggesting that the simultaneous presence of the three domains is essential for high-affinity RNA-binding. ii) Deletion of the Ala-rich auxiliary domain did hardly affect RNA-binding. Thus, this structural subunit may not be involved in RNA interaction. iii) Deletion of the acidic auxiliary domain improved the RNA-binding suggesting a regulatory role for this structural motif. iv) The isolated RNA-binding domains did not exhibit sizeable RNA-binding affinities. Taken together these data suggest that a cooperative interaction of the three qRRMs is required for high affinity RNA-binding.

CIZ1 in Xist seeded assemblies at the inactive X chromosome.

There is growing evidence that X-chromosome inactivation is driven by phase-separated supramolecular assemblies. However, among the many proteins recruited to the inactive X chromosome by Xist long non-coding RNA, so far only a minority (CIZ1, CELF1, SPEN, TDP-43, MATR3, PTBP1, PCGF5) have been shown to form Xist-seeded protein assemblies, and of these most have not been analyzed in detail. With focus on CIZ1, here we describe 1) the contribution of intrinsically disordered regions in RNA-dependent protein assembly formation at the inactive X chromosome, and 2) enrichment, distribution, and function of proteins within Xist-seeded assemblies.

Impact of germination on structural, physicochemical, techno-functional, and digestion properties of desi chickpea (Cicer arietinum L.) flour.

Germination significantly increased the nutrient composition, total phenolic content, and antioxidant activity of the chickpea flour cultivars studied (GNG-469 and GNG-1581). Protein and starch digestibilities were significantly improved in germinated chickpea flour. Germinated chickpea flour formed gels that had a shear modulus about 60% that of the non-germinated ones. X-ray diffraction analysis indicated that both A and B type polymorphs were present in the chickpea flour cultivars. Germination significantly reduced the relative crystallinity of the chickpea flour cultivars, from around 33 to 27% for GNG 469 and around 30 to 25% for GNG 1581. Protein secondary structures showed increase in ß-sheets and random coils content in chickpea cultivars during germination. Phenolic acid profiling showed a decrease in the concentration of ellagic, p-coumaric, p-hydroxybenzoic, and caffeic acids but an increase in gallic, p-coumaric, and ferulic acids after germination.

Shifting archetype to nature's hidden gems: from sources, purification to uncover the nutritional potential of bioactive peptides.

Contemporary scientific findings revealed that our daily food stuffs are enriched by encrypted bioactive peptides (BPs), evolved by peptide linkage of amino acids or encrypted from the native protein structures. Remarkable to these BPs lies in their potential health benefiting biological activities to serve as nutraceuticals or a lead addition to the development of functional foods. The biological activities of BPs vary depending on the sequence as well as amino acid composition. Existing database records approximately 3000 peptide sequences which possess potential biological activities such as antioxidants, antihypertensive, antithrombotic, anti-adipogenics, anti-microbials, anti-inflammatory, and anti-cancerous. The growing evidences suggest that BPs have very low toxicity, higher accuracy, less tissue accretion, and are easily degraded in the disposed environment. BPs are nowadays evolved as biologically active molecules with potential scope to reduce microbial contamination as well as ward off oxidation of foods, amend diverse range of human diseases to enhance the overall quality of human life. Against the clinical and health perspectives of BPs, this review aimed to elaborate current evolution of nutritional potential of BPs, studies pertaining to overcome limitations with respect to special focus on emerging extraction, protection and delivery tools of BPs. In addition, the nano-delivery mechanism of BP and its clinical significance is detailed. The aim of current review is to augment the research in the field of BPs production, identification, characterisation and to speed up the investigation of the incredible potentials of BPs as potential nutritional and functional food ingredient.

Nutritional and bioactive characteristics of buckwheat, and its potential for developing gluten-free products: An updated overview.

In the present era, food scientists are concerned about exploiting functional crops with nutraceutical properties. Buckwheat is one of the functional pseudocereals with nutraceutical components used in the treatment of health-related diseases, malnutrition, and celiac diseases. As a preferred diet as a gluten-free product for celiac diseases, buckwheat is a good source of nutrients, bioactive components, phytochemicals, and antioxidants. The general characteristics and better nutritional profile of buckwheat than other cereal family crops were highlighted by previous investigations. In buckwheats, bioactive components like peptides, flavonoids, phenolic acids, d-fagomine, fagopyritols, and fagopyrins are posing significant health benefits. This study highlights the current knowledge about buckwheat and its characteristics, nutritional constituents, bioactive components, and their potential for developing gluten-free products to target celiac people (1.4% of the world population) and other health-related diseases.

Insight about the biochemical composition, postharvest processing, therapeutic potential of Indian gooseberry (amla), and its utilization in development of functional foods-A comprehensive review.

Indian gooseberry/Amla (Emblica officinalis Gaertn. syn. Phyllanthus emblica L) has an amazing nutritional profile and is a reservoir of biologically active compounds which have potential health benefits and are regarded as a remedy for lethal diseases. The unique features of amla, conferred by their bioactive components, have extended future prospects about their usage for useful effects on human nutrition and health globally. With the rapidly growing popularity of this unique therapeutic fruit, it is important to have comprehensive knowledge of this fruit. The current review article presents the nutritional profile, bioactive components, antioxidant and antimicrobial properties, and postharvest processing of amla fruit. Moreover, studies related to therapeutic properties of amla and its utilization in development of functional foods have been presented in this review. E. officinalis is a promising source of bioactive compounds which showed varied potential in the management of a number of human ailments which has been proven through various studies. Therefore, amla should be taken in the regular diet, thereby utilizing its potential health benefits. PRACTICAL APPLICATIONS: Amla (Indian gooseberry), as source of natural bioactive compounds, has a great potential application in improving the status of human nutrition and health. The utilization of amla extract has various biological effects, like antimicrobial, antioxidant, gastroprotective, anticancer, hepatoprotective, cardioprotective, radioprotective, anti-inflammatory, antidiabetic, neuroprotective, and immunomodulatory effect, owing to its bioactive components. The use of amla extract has recently increased in food, pharmaceutical, and cosmetic products to replace synthetic antioxidants which have inherent harmful health effects. The review report will provide information on bioactive components, therapeutic properties, utilization of amla in the development of future functional foods, and postharvest processing of amla, which will provide critical information to researchers all over the world.

Male guanine-rich RNA sequence binding factor 1 knockout mice (Grsf1-/-) gain less body weight during adolescence and adulthood.

The guanine-rich RNA sequence binding factor 1 (GRSF1) is an RNA-binding protein of the heterogenous nuclear ribonucleoprotein H/F (hnRNP H/F) family that binds to guanine-rich RNA sequences forming G-quadruplex structures. In mice and humans there are single copy GRSF1 genes, but multiple transcripts have been reported. GRSF1 has been implicated in a number of physiological processes (e.g. embryogenesis, erythropoiesis, redox homeostasis, RNA metabolism) but also in the pathogenesis of viral infections and hyperproliferative diseases. These postulated biological functions of GRSF1 originate from in vitro studies rather than complex in vivo systems. To assess the in vivo relevance of these findings, we created systemic Grsf1-/- knockout mice lacking exons 4 and 5 of the Grsf1 gene and compared the basic functional characteristics of these animals with those of wildtype controls. We found that Grsf1-deficient mice are viable, reproduce normally and have fully functional hematopoietic systems. Up to an age of 15 weeks they develop normally but when male individuals grow older, they gain significantly less body weight than wildtype controls in a gender-specific manner. Profiling Grsf1 mRNA expression in different mouse tissues we observed high concentrations in testis. Comparison of the testicular transcriptomes of Grsf1-/- mice and wildtype controls confirmed near complete knock-out of Grsf1 but otherwise subtle differences in transcript regulations. Comparative testicular proteome analyses suggested perturbed mitochondrial respiration in Grsf1-/- mice which may be related to compromised expression of complex I proteins. Here we present, for the first time, an in vivo complete Grsf1 knock-out mouse with comprehensive physiological, transcriptomic and proteomic characterization to improve our understanding of the GRSF1 beyond in vitro cell culture models.

Prion-like domains drive CIZ1 assembly formation at the inactive X chromosome.

CIZ1 forms large assemblies at the inactive X chromosome (Xi) in female fibroblasts in an Xist lncRNA-dependent manner and is required for accurate maintenance of polycomb targets genome-wide. Here we address requirements for assembly formation and show that CIZ1 undergoes two direct interactions with Xist, via independent N- and C-terminal domains. Interaction with Xist, assembly at Xi, and complexity of self-assemblies formed in vitro are modulated by two alternatively spliced glutamine-rich prion-like domains (PLD1 and 2). PLD2 is dispensable for accumulation at existing CIZ1-Xi assemblies in wild-type cells but is required in CIZ1-null cells where targeting, assembly, and enrichment for H3K27me3 and H2AK119ub occur de novo. In contrast, PLD1 is required for both de novo assembly and accumulation at preexisting assemblies and, in vitro, drives formation of a stable fibrillar network. Together they impart affinity for RNA and a complex relationship with repeat E of Xist. These data show that alternative splicing of two PLDs modulates CIZ1's ability to build large RNA-protein assemblies.

Expression Regulation, Protein Chemistry and Functional Biology of the Guanine-Rich Sequence Binding Factor 1 (GRSF1).

In eukaryotic cells RNA-binding proteins have been implicated in virtually all post-transcriptional mechanisms of gene expression regulation. Based on the structural features of their RNA binding domains these proteins have been divided into several subfamilies. The presence of at least two RNA recognition motifs defines the group of heterogenous nuclear ribonucleoproteins H/F and one of its members is the guanine-rich sequence binding factor 1 (GRSF1). GRSF1 was first described 25 years ago and is widely distributed in eukaryotic cells. It is present in the nucleus, the cytoplasm and in mitochondria and has been implicated in a variety of physiological processes (embryogenesis, erythropoiesis, redox homeostasis, RNA metabolism) but also in the pathogenesis of various diseases. This review summarizes our current understanding on GRSF1 biology, critically discusses the literature reports and gives an outlook of future developments in the field.

Application of new technologies in decontamination of mycotoxins in cereal grains: Challenges, and perspectives.

Emerging decontamination technologies have been attracted considerable attention to address the consumers' demand for high quality and safe food products. As one of the important foods in the human diet, cereals are usually stored for long periods, resulting in an increased risk of contamination by different hazards. Mycotoxins comprise one of the significant contaminants of cereals that lead to enormous economic losses to the industry and threats to human health. While prevention is the primary approach towards reducing human exposure to mycotoxins, decontamination methods have also been developed as complementary measures. However, some conventional methods (chemical treatments) do not fulfill industries' expectations due to limitations like safety, efficiency, and the destruction of food quality attributes. In this regard, novel techniques have been proposed to food to comply with the industry's demand and overcome conventional methods' limitations. Novel techniques have different efficiencies for removing or reducing mycotoxins depending on processing conditions, type of mycotoxin, and the food matrix. Therefore, this review provides an overview of novel mycotoxin decontamination technologies such as cold plasma, irradiation, and pulse light, which can be efficient for reducing mycotoxins with minimum adverse effects on the quality and nutritional properties of produce.

Identification of the COMM-domain containing protein 1 as specific binding partner for the guanine-rich RNA sequence binding factor 1.

BACKGROUND: The guanine-rich RNA sequence binding factor 1 (GRSF1) is an RNA-binding protein of the hnRNP H/F family, which has been implicated in erythropoiesis, regulation of the redox homeostasis, embryonic brain development, mitochondrial function and cellular senescence. The molecular basis for GRSF1-RNA interaction has extensively been studied in the past but for the time being GRSF1 binding proteins have not been identified. METHODS: To search for GRSF1 binding proteins we first employed the yeast two-hybrid system and screened a cDNA library of human fetal brain for potential GRSF1 binding proteins. Subsequently, we explored the protein-protein-interaction of the recombiant proteins, carried out immunoprecipitation experiments to confirm the interaction of the native proteins in living cells and performed truncation studies to identify the protein-binding motif of GRSF1. RESULTS: Using the yeast two-hybrid system we identified the COMM-domain containing protein 1 (COMMD1) as specific GRSF1 binding protein and in vitro truncation studies suggested that COMMD1 interacts with the alanine-rich domain of GRSF1. Co-immunoprecipitation strategies indicated that COMMD1-GRSF1 interaction was RNA independent and also occurred in living cells expressing the two native proteins. CONCLUSION: In mammalian cells the COMM-domain containing protein 1 (COMMD1) specifically interacts with the Ala-rich domain of GRSF1 in an RNA-independent manner. GENERAL SIGNIFICANCE: This is the first report describing a specific GRSF1 binding protein.

Grsf1-induced translation of the SNARE protein Use1 is required for expansion of the erythroid compartment.

Induction of cell proliferation requires a concomitant increase in the synthesis of glycosylated lipids and membrane proteins, which is dependent on ER-Golgi protein transport by CopII-coated vesicles. In this process, retrograde transport of ER resident proteins from the Golgi is crucial to maintain ER integrity, and allows for anterograde transport to continue. We previously showed that expression of the CopI specific SNARE protein Use1 (Unusual SNARE in the ER 1) is tightly regulated by eIF4E-dependent translation initiation of Use1 mRNA. Here we investigate the mechanism that controls Use1 mRNA translation. The 5'UTR of mouse Use1 contains a 156 nt alternatively spliced intron. The non-spliced form is the predominantly translated mRNA. The alternatively spliced sequence contains G-repeats that bind the RNA-binding protein G-rich sequence binding factor 1 (Grsf1) in RNA band shift assays. The presence of these G-repeats rendered translation of reporter constructs dependent on the Grsf1 concentration. Down regulation of either Grsf1 or Use1 abrogated expansion of erythroblasts. The 5'UTR of human Use1 lacks the splice donor site, but contains an additional upstream open reading frame in close proximity of the translation start site. Similar to mouse Use1, also the human 5'UTR contains G-repeats in front of the start codon. In conclusion, Grsf1 controls translation of the SNARE protein Use1, possibly by positioning the 40S ribosomal subunit and associated translation factors in front of the translation start site.