In vitro antioxidant and antimicrobial activities of clove extract and its effectiveness in bio-composite film on storage stability of goat meat balls.

The aim of this study was to find out clove extract's antimicrobial and antioxidant properties, as well as its efficacy as a bioactive ingredient in the development of bio-composite films to increase the storage stability of goat meat balls stored at 4 ± 1°C. The clove extracts (CLEs) were prepared in ethanol, hydroethanol (1:1), and water and evaluated for antioxidant and antimicrobial potential. In vitro assays of CLEs revealed more susceptibility for gram-positive bacteria than gram-negative bacteria. Among the different extracts, the clove ethanol extract (CLEE) had the highest antimicrobial activity against tested microorganisms as well as total phenolics (1.14 mg GAE/g), flavonoids (8.50 µg catechin/g), and DPPH assay (39.59%). Further, the concentration-dependent effect of CLEE (p < 0.05) on thickness and color values and antimicrobial properties of the bio-composite film were observed. The storage qualities of the product T1 (with film; 450 µl CLEE) such as pH (6.45 ± 0.01), TBARS (0.87 ± 0.06 mg malonaldehyde/kg) value, free fatty acid (0.193 ± 0.001% oleic acid), total mesophilic count (4.98 ± 0.05 log10 CFU/g), and sensory attributes (overall acceptability score: 5.67 on 8-point scale) were better (p < 0.05) than T0 (without film; control) on day 20 of storage. Thus, the ethanolic clove extract has a superior antioxidant and antimicrobial potential. Its inclusion in the bio-composite film prolonged the storage stability of goat meat balls by controlling lipid oxidation and microbial growth. Practical Application Today's consumers are more attracted towards meat products added with natural ingredients having preservative effects. Clove extract is a classic example of a natural preservative and has excellent antimicrobial and antioxidant potential. The present study revealed that by wrapping the ethanolic clove extract-based bio-composite film on goat meat balls extended the storage stability of the product due to controlled lipid oxidation and microbial growth. Thus, such bio-composite films can be successfully applied on goat meat balls that function as a antimicrobial packaging for providing optimum organoleptic quality and better shelf life.

Small Molecule Screening Discovers Compounds that Reduce FMRpolyG Protein Aggregates and Splicing Defect Toxicity in Fragile X-Associated Tremor/Ataxia Syndrome.

Expansion of CGG trinucleotide repeats in 5' untranslated region of the FMR1 gene is the causative mutation of neurological diseases such as fragile X syndrome (FXS), fragile X-associated tremor/ataxia syndrome (FXTAS), and ovarian disorder such as fragile X-associated primary ovarian insufficiency (FXPOI). CGG repeats containing FMR1 transcripts form the toxic ribonuclear aggregates, abrupt pre-mRNA splicing, and cause repeat-associated non-AUG translation, leading to the disease symptoms. Here, we utilized a small molecule library of ~ 250,000 members obtained from the National Cancer Institute (NCI) and implemented a shape-based screening approach to identify the candidate small molecules that mitigate toxic CGG RNA-mediated pathogenesis. The compounds obtained from screening were further assessed for their affinity and selectivity towards toxic CGG repeat RNA by employing fluorescence-binding experiment and isothermal calorimetry titration assay. Three candidate molecules B1, B4, and B11 showed high affinity and selectivity for expanded CGG repeats RNA. Further, NMR spectroscopy, gel mobility shift assay, CD spectroscopy, UV-thermal denaturation assay, and molecular docking affirmed their high affinity and selectivity for toxic CGG RNAs. Next, these lead compounds selectively improved the pre-mRNA alternative splicing defects with no perturbation in global splicing efficacy and simultaneously reduced the FMR1polyG protein aggregate formation without affecting the downstream expression of the gene. Taken together these findings, we addressed compound B1, B4, and B11 as potential lead molecules for developing promising therapeutics against FXTAS. Herein, this study, we have utilized shape similarity approach to screen the NCI library and found out the potential candidate which improves the pre-mRNA splicing defects and reduces FMR1polyG aggregations.

Patterns of Physical Activity Among University Students and Their Perceptions About the Curricular Content Concerned With Health: Cross-sectional Study.

BACKGROUND: University students are at risk of losing their focus on maintaining healthy levels of physical activity because of their engagements with curricular and cocurricular activities. In India, the physical activity levels of the adult population have been reported to be declining in the recent years. However, studies focusing on university students pertaining to their physical activity are lacking in the Indian context. Moreover, a question that has not been properly addressed is the following: "do the curricula in higher education promote physical activity?" OBJECTIVE: Our paper aims at describing the physical activity levels of the students in a large public-funded central university located in northern India. The study also aims at capturing the student perceptions about the emphasis they receive on leading a physically active lifestyle during their routine curricular activities. METHODS: This is a cross-sectional descriptive study and uses International Physical Activity Questionnaire-Long Form to record physical activity among 4586 students. Stratified sampling method was used to enroll the students from each stream (faculty). Out of 30,667 students, about 15% were included from each faculty. The study was conducted between 2016 and 2019. To capture the student perceptions, we used a newly developed 5-item scale. RESULTS: From a total of 4586 participants in the study, 2828 (61.7%) were male and 1758 (38.3%) were female students. The mean age of our sample was 22.34 (SD 3.12) years. Our results indicate that about 14.5% (n=666) of all students in the study fall under the "Inactive" category. Furthermore, the perception about the curricular content pertaining to physical activity varied widely between the students of different streams. CONCLUSIONS: Our sample reported a better physical activity pattern in comparison to the reported overall physical activity levels of the adult population of India. Our results also suggest that health-related topics are inadequately represented in many of the streams of higher education in the university.

Materialization of novel composite bio-based active edible film functionalized with essential oils on antimicrobial and antioxidative aspect of chicken nuggets during extended storage.

The present study was aimed to develop Manihot esculenta and Carrageenan bio-based composite active film functionalized with anise, caraway, and nutmeg essential oils (EOs) and to assess the shelf life of chicken nuggets wrapped with the developed film at refrigeration storage. Overall, the Minimum Inhibitory Concentration (MIC) values of the three EOs ranged from 0.4 to 0.8% v/v of which nutmeg EO was found most effective. Incorporation of EOs in the film resulted in significant (P ˂ 0.05) decrease in tensile strength and water activity while elongation at break was significantly (P ˂ 0.05) increased. No significant (P ˃ 0.05) changes in thickness and Water Vapor Transmission Rate (WVTR) were observed. Based on physicomechanical and sensory evaluation, films incorporated with 0.5, 1, and 1% concentration of anise, nutmeg, and caraway EO were selected. Chicken nuggets overwrapped with aforementioned films were stored aerobically at refrigeration temperature (4 ± 1 °C) to evaluate antimicrobial, antioxidant, and sensory characteristics. The result indicated that pH, peroxide, free fatty acid (FFA), and thiobarbituric acide (TBA) value of treatments were significantly (P ˂ 0.05) lower than controls however significantly (P < 0.05) higher DPPH activity was observed in all treatments. The total plate count, psychrophilic count and, yeast and mold count were also significantly (P ˂ 0.01) lower in treatment groups and were within the permissible limits. The treated samples were well acceptable during whole storage period of 15 days. The application of composite, active edible bio-based film was found proficient in confining product quality attributes throughout storage. PRACTICAL APPLICATION: The majority of films used for packaging of meat and meat products are derived from synthetic "plastic" materials. The demerits associated with plastics have eventually led to explore natural alternatives such as edible films. The composite-active bio-based films have a huge potential to be molded for specific film properties based on requirements of product-specific packaging conditions.

Curcumin Regulates the r(CGG)exp RNA Hairpin Structure and Ameliorate Defects in Fragile X-Associated Tremor Ataxia Syndrome.

Fragile X-associated tremor ataxia syndrome is an untreatable neurological and neuromuscular disorder caused by unstable expansion of 55-200 CGG nucleotide repeats in 5' UTR of Fragile X intellectual disability 1 (FMR1) gene. The expansion of CGG repeats in the FMR1 mRNA elicits neuronal cell toxicity through two main pathogenic mechanisms. First, mRNA with CGG expanded repeats sequester specific RNA regulatory proteins resulting in splicing alterations and formation of ribonuclear inclusions. Second, repeat-associated non-canonical translation (RANT) of the CGG expansion produces a toxic homopolymeric protein, FMRpolyG. Very few small molecules are known to modulate these pathogenic events, limiting the therapeutic possibilities for FXTAS. Here, we found that a naturally available biologically active small molecule, Curcumin, selectively binds to CGG RNA repeats. Interestingly, Curcumin improves FXTAS associated alternative splicing defects and decreases the production and accumulation of FMRpolyG protein inclusion. Furthermore, Curcumin decreases cell cytotoxicity promptly by expression of CGG RNA in FXTAS cell models. In conclusion, our data suggest that small molecules like Curcumin and its derivatives may be explored as a potential therapeutic strategy against the debilitating repeats associated neurodegenerative disorders.

Exploring the Potential of Small Molecule-Based Therapeutic Approaches for Targeting Trinucleotide Repeat Disorders.

In recent years, neurological disorders have globally become a leading cause of disability and death. Neurological disorders are very common in both high- and low-income countries, and the number of patients is predicted to increase in the coming decades. Disorders caused by the expanded trinucleotide repeats (CAG, CGG, CCG, CTG, CUG, GAA, and GCN) in the genome, also described as trinucleotide repeat expansion disorders (TREDs), comprise of the major class of neurological diseases. Various TREDs have different modes of pathogenesis, but the severity and time of onset of disease depends on the trinucleotide repeat numbers. Numerous therapeutic strategies, including symptomatic treatment, blockage of mutant protein synthesis, targeting the toxic protein aggregates and degradation of RNA transcripts have been developed for the treatment of these diseases. However, various limitations to these therapeutic strategies have been reported, and therefore, researchers are exploring different avenues of therapeutics development. One of the recent developments include targeting the expanded repeats with small molecules. Small molecule binds with the secondary/tertiary structure of RNA (like bulges, loops, and hairpins) irrespective of its sequences. Altogether, small molecule-based therapeutics may have the advantage over others to be able to overcome the hurdles of the blood-brain barrier, poor absorption, and allergic reactions. In this review, we have summarized various TREDs and envisage the potential of small molecule-based therapeutics for targeting these hitherto incurable neurological disorders.

Piperine Modulates Protein Mediated Toxicity in Fragile X-Associated Tremor/Ataxia Syndrome through Interacting Expanded CGG Repeat (r(CGG)exp) RNA.

An expansion of CGG tandem repeats in the 5' untranslated region (5'-UTR) of fragile X mental retardation 1 (FMR1) gene causes fragile X-associated tremor/ataxia syndrome (FXTAS). The transcripts of these expanded repeats r(CGG)exp either form RNA foci or undergo the repeat-associated non-ATG (RAN) translation that produces toxic homopolymeric proteins in neuronal cells. The discovery of small molecule modulators that possess a strong binding affinity and high selectivity to these toxic expanded repeats RNA could be a promising therapeutic approach to cure the expanded repeat-associated neurological diseases. Therefore, here we sought to test the therapeutic potential of a natural alkaloid, piperine, by assessing its ability to bind and neutralize the toxicity of r(CGG)exp RNA motif. To accomplish this first, we have determined the affinity of piperine to r(CGG)exp RNA using fluorescence-based binding assay and isothermal titration calorimetry assay. These assays showed that piperine forms a thermodynamically favorable interaction with r(CGG)exp RNA with high selectivity to the G-rich RNA motif. Interaction of piperine with r(CGG)exp motif was further validated using several biophysical techniques such as CD, CD melting, NMR spectroscopy, and gel retardation assay. Moreover, piperine was also found to be effective for improving the r(CGG)exp associated splicing defects and RAN translation in a FXTAS cell model system. Our results effectively provided the evidence that piperine strongly interacts with r(CGG)exp RNA and could be used as a suitable candidate for therapeutic development against FXTAS.

Myricetin Reduces Toxic Level of CAG Repeats RNA in Huntington's Disease (HD) and Spino Cerebellar Ataxia (SCAs).

Huntington's disease (HD) is a neurodegenerative disorder that is caused by abnormal expansion of CAG repeats in the HTT gene. The transcribed mutant RNA contains expanded CAG repeats that translate into a mutant huntingtin protein. This expanded CAG repeat also causes mis-splicing of pre-mRNA due to sequestration of muscle blind like-1 splicing factor (MBNL1), and thus both of these elicit the pathogenesis of HD. Targeting the onset as well as progression of HD by small molecules could be a potent therapeutic approach. We have screened a set of small molecules to target this transcript and found Myricetin, a flavonoid, as a lead molecule that interacts with the CAG motif and thus prevents the translation of mutant huntingtin protein as well as sequestration of MBNL1. Here, we report the first solution structure of the complex formed between Myricetin and RNA containing the 5'CAG/3'GAC motif. Myricetin interacts with this RNA via base stacking at the AA mismatch. Moreover, Myricetin was also found reducing the proteo-toxicity generated due to the aggregation of polyglutamine, and further, its supplementation also improves neurobehavioral deficits in the HD mouse model. Our study provides the structural and mechanistic basis of Myricetin as an effective therapeutic candidate for HD and other polyQ related disorders.

Quality characteristics of low-fat chicken nuggets: effect of common salt replacement and added bottle gourd (Lagenaria siceraria L.).

BACKGROUND: There is growing demand for the meat products having healthier characteristics. In an endeavour to develop low-salt, low-fat and high-fibre chicken nuggets an investigation was carried out to observe the effects of partial replacement (40%) of sodium chloride in pre-standardised low-fat chicken nuggets (Control, 20 g kg⁻¹ NaCl) with a salt substitute blend as well as incorporation of bottle gourd (Lagenaria siceraria L.) in the resulting low-salt, low-fat products at three different levels, i.e. 50, 75 and 100 g kg⁻¹ (Treatments, 12 g kg⁻¹ NaCl) on the various quality attributes. RESULTS: Sodium chloride replacement decreased (P < 0.01) emulsion and product pH, cooking yield, moisture, ash, yellowness, hue value and textural properties. pH values, moisture and dietary fibre increased (P < 0.01) while cooking yield, % protein, textural properties and total cholesterol were decreased with the incorporation of bottle gourd in low-salt, low-fat nuggets. Sensory attributes of the product were not affected with salt replacement; however, inclusion of bottle gourd at higher levels decreased (P < 0.05) flavour and texture scores. CONCLUSION: The results suggest that low-salt, low-fat and high-fibre chicken nuggets can be developed with the use of a salt substitute blend and bottle gourd without affecting their acceptability.

Low-sodium meat products: retaining salty taste for sweet health.

There is a positive correlation between excessive intake of sodium and incidence of hypertension. As diet is the main source of sodium, awareness among people regarding its possible role upon health has driven demand for various low sodium foods including meat products. Meat products contribute a significant amount of dietary sodium, thus maligning their own image. However, this is not an easy task as common salt affects taste and flavor, functional attributes, stability, and food safety of meat products. The various properties such as taste and flavor, binding, as well as microbiological characteristics should be given due care while developing low salt meat products and accordingly different approaches have been proposed for processing of such products. Potassium chloride has been mostly used to replace sodium; however, a number of other salts, flavor enhancers, bitter blockers and water, as well as fat binders have also been attempted either alone or in different combinations. A number of low sodium meat products have been developed but their economy and consumer acceptability are the major concerns needing proper attention. In future it is anticipated that these challenges would be overcome to provide well acceptable and cost-effective healthier meat products to the consumers.

Dietary fibre as functional ingredient in meat products: a novel approach for healthy living - a review.

There is a rapid change in our overall lifestyle due to impact of globalization. Every day hasty life has forced consumers to be dependent upon fast foods, which contain meagre amount of dietary fibre. Non-starch polysaccharides and resistant oligosaccharides, lignin, substances associated with NSP and lignin complex in plants, other analogous carbohydrates, such as resistant starch and dextrins, and synthesized carbohydrate compounds, like polydextrose are categorized as dietary fibre. They are mostly concentrated in cereals, pulses, fruits and vegetables. It has been proclaimed that daily dietary fibre intake helps in prevention of many nutritional disorders like gut related problems, cardiovascular diseases, type 2 diabetes, certain types of cancer and obesity. Meat is generally lacking this potential ingredient, which could be incorporated while products processing to make them more healthful. Various fibre rich sources have been attempted in different products attributed to their technological and health benefits and many are in the queue to be used in a variety of meat products. Selection of appropriate fibre rich ingredients and their proper incorporation can improve health image of meat products.