The attempt of this review article is to determine the impact of nuclear and mitochondrial damages on the propagation of cancer incidences. This review has advanced our understanding to altered genes and their relevant cancerous proteins. The progressive raising effects of free reactive oxygen species ROS and toxicogenic compounds contributed to significant mutation in nuclear and mitochondrial DNA where the incidence of gastric cancer is found to be linked with down regulation of some relevant genes and mutation in some important cellular proteins such as AMP-18 and CA-11. Thereby, the resulting changes in gene mutations induced the apparition of newly polymorphisms eventually leading to unusual cellular expression to mutant proteins. Reduction of these apoptotic growth factors and nuclear damages is increasingly accepted by cell reactivation effect, enhanced cellular signaling and DNA repairs. Acetylation, glycation, pegylation and phosphorylation are among the molecular techniques used in DNA repair for rectifying mutation incidences. In addition, the molecular labeling based fluorescent materials are currently used along with the bioconjugating of signal molecules in targeting disease translocation site, particularly cancers and tumors. These strategies would help in determining relevant compounds capable in overcoming problems of down regulating genes responsible for repair mechanisms. These issues of course need interplay of both proteomic and genomic studies often in combination of molecular engineering to cible the exact expressed gene relevant to these cancerous proteins.
Mitragynine is the main psychoactive compound of Mitragyna speciosa Korth. (kratom). This alkaloid could render psychotropic effects and is often misused as a substitute for commercial drugs. Nowadays, the increasing popularity of kratom has led to the development of a rapid and effective detection method. The detection of mitragynine in a biological sample such as urine requires a highly sensitive and specific method due to the complex nature of mitragynine in urine. Enzyme-linked immunosorbent assay (ELISA) is well known as a rapid screening method for biological samples. In this study, a competitive indirect ELISA was successfully developed using MG-22-OCH3 IgG as a detection antibody for mitragynine in human urine. The mitragynine immunoassay showed a limit of detection and a limit of quantification of 0.412 and 1.25 μg/mL, respectively. The measurement range was between 0.01 and 100.0 μg/mL, with a minimal inhibition (IC50) value of 0.152 μg/mL. The developed ELISA was validated using a gold method such as high-performance liquid chromatography-mass spectrometry (HPLC-MS). The percentage of recovery and the coefficient of variation (CV) for the ELISA and LCMS/MS analyses were 84.0-95.70%, 99.20-112.0%, 7.69-9.78%, and 2.86-6.62%, respectively. This indicates that the developed ELISA is a reliable method that can be used as a rapid approach for quantifying mitragynine content in biological samples.
In the ongoing pursuit of novel and efficient NLO materials, the potential of alkali metal-doped {6}cycloparaphenylene ({6}CPP) and methylene bridged {6} cycloparaphenylene (MB{6}CPP) nanohoops as excellent NLO candidates has been explored. The geometric, electronic, linear, and nonlinear optical properties of designed systems have been investigated theoretically. All the nanohoops demonstrated thermodynamic stability, with remarkable interaction energies reaching up to -1.39 eV (-0.0511 au). Notably, the introduction of alkali metals led to a significant reduction in the HOMO-LUMO energy gaps, with values as low as 2.92 eV, compared to 6.80 eV and 6.06 eV for undoped {6}CPP and MB{6}CPP, respectively. Moreover, the alkali metal-doped nanohoops exhibited exceptional NLO response, with the K@r6-{6}CPP complex achieving the highest first hyperpolarizability of 56,221.7 × 10-30 esu. Additionally, the frequency-dependent first hyperpolarizability values are also computed at two commonly used wavelengths of 1550 nm and 1907 nm, respectively. These findings highlight the potential of designed nanohoops as promising candidates for advanced NLO materials with high-tech applications.
Gamma-aminobutyric acid (γ-ABA) can be produced by various microorganisms including bacteria, fungi and yeasts using enzymatic bioconversion, microbial fermentation or chemical hydrolysis. Regenerating conjugated glycerol-amines is valid by the intervention of microbial cyclooxygenase [COX] and lipooxygenase [LOX] enzymes produced via lactobacillus bacteria (LAB) as successor enzymes to glutamate decarboxylases (GAD). Therefore, the aim of this review is to provide an overview on γ-ABA production, and microbiological achievements used in producing this signal molecule based on those fermenting enzymes. The formation of aminoglycerides based conjugated γ-ABA is considered the key substances in controlling the host defense against pathogens and is aimed in increasing the neurotransmission effects and in suppressing further cardiovascular diseases.
Rice bran protein concentrates (RBPC) were extracted using mild alkaline solvents (pH: 8, 9, 10). The physicochemical, thermal, functional, and structural aspects of freeze-drying (FD) and spray-drying (SD) were compared. FD and SD of RBPC had porous and grooved surfaces, with FD having non-collapsed plates and SD being spherical. Alkaline extraction increases FD's protein concentration and browning, whereas SD inhibits browning. According to amino acid profiling, RBPC-FD9's extraction optimizes and preserves amino acids. A tremendous particle size difference was prominent in FD, thermally stable at a minimal maximum of 92 °C. Increased pH extraction gives FD greater exposal surface hydrophobicity and positively relates to denaturation enthalpy. Mild pH extraction and drying significantly impacted solubility, improved emulsion properties, and foaming properties of RBPC as observed in acidic, neutral, and alkaline environments. RBPC-FD9 and RBPC-SD10 extracts exhibit outstanding foaming and emulsion activity in all pH conditions, respectively. Appropriate drying selection, RBPC-FD or SD potentially employed as foaming/emulsifier agent or meat analog.
There is an increasing demand for natural food preservatives due to consumers' concern on the negative effects of chemical preservatives in food products. Nisin (bacteriocin) is an effective food biopreservative that has been approved globally. However, its low yield proves to be a limiting factor and must be addressed to meet the increasingly high demand from the food industry. The present work thus investigated the effects of individual and combined fermentation factors on Lactococcus lactis ATCC 11454 growth and nisin activity using the one-factor-at-a-time (OFAT) method. The level of each factor that gave the highest nisin production was then selected and combined to further improve its activity. The best combined conditions for highest cell growth and nisin activity were 30 °C, pH 6.0, and mild agitation with the addition of 1.0% w/v glucose, 1.0% w/v skim milk, and 0.5% v/v Tween 20. This increased nisin production by 22.7% as compared to control (basic condition). The present work provided critical information on the relationship between fermentation conditions, growth, and nisin activity of L. lactis ATCC 11454 that could be explored to understand the potential and limitation of the strain. This fermentation strategy can also serve as a benchmark to further enhance the production of bacteriocin or other biopreservative compounds.
Three novel peptide sequences YGIKVGYAIP, GGIF, and GIFE from papain-generated protein hydrolysate of palm kernel cake proteins were used for stability study against ACE, ACE-inhibition kinetics, and molecular docking studies. Results showed that peptide YGIKVGYAIP was degraded, and its ACE-inhibitory activity decreased after 3 h pre-incubation with ACE, while peptides GGIF and GIFE were resistant. However, although the ACE-inhibitory activity of GIFE increased during this time, the ACE inhibitory activity of GGIF decreased after pre-incubation with ACE, indicating that peptide. YGIKVGYAIP and GGIF are substrate-type, whereas GIFE is a true-inhibitor type. Peptide YGIKVGYAIP showed the lowest Ki (0.054 mM) in the inhibition kinetics study compared to GGIF and GIFE, with Ki of 1.27 m M and 18 mM, respectively. In addition, YGIKVGYAIP revealed the lowest Km and Vmax and higher CE in different peptide concentrations, implying that the enzyme catalysis decreased, and peptides had some binding affinity to the enzyme in lower concentrations, which led to reduced catalytic ability. Furthermore, YGIKVGYAIP showed the lowest docking score of -14.733 and 21 interactions with tACE, while GGIF revealed the higher docking score of -8.006 with 15 interactions with tACE.
Glutens are potential proteins with multifunctional therapeutic effects. Their covalence network structures with and without protease inhibitors are expected to enhance or to serve further properties and further technological points such as increased bioactive surfaces, gelatinization, gelation and pasting properties. The depletion of the allergic peptide sequences of gluten proteins comprising sometimes protease inhibitors are valid via the enzymatic ingestion using proteolytic enzymes that might enhance these functional and technological processes by producing active peptides having osmoregulation and regular glass transitions, surface activity for coating and encapsulation properties. In addition to further therapeutic functions such as immunoregulatory, antithrombin and opioidal activities, particularly in eradicating most of the free radicals, suppressing diabetes Mellitus II complications and inhibiting angiotensin converting enzyme cardiovascular growth diseases.
Ergogenic property is the ability to enhance capacity for physical activities through efficient production of energy and is potentially beneficial in weight management for the obese. In this study, ergogenic property of Morinda citrifolia leaf's extract (MCL) was evaluated using AMP-activated protein kinase (AMPK) activity and high fat diet-induced obese rats. Findings from the study showed that MCL demonstrated ergogenic activity via enhancement of AMPK activity using L6 skeletal muscle cell line. Interestingly, the result also revealed that rats treated with the intermediate dosage of MCL experienced the lowest % weight gain. The rats fed the highest dose of 200 mg/kg BW MCL demonstrated the longest swimming time of approximately three times that of green tea and caffeine-fed rats. The highest dose fed rats were also found to have lower glucose and lactate levels, suggesting that energy metabolism was more effective in these rats. In addition, lactate dehydrogenase and creatinine kinase activities, the muscle injury indicators, were found to be the lowest in rats fed the highest MCL dose. The same effect was not seen in rats fed either caffeine or green tea, indicating that MCL treatment is may be protective of the rats' muscles. It was also shown that MCL consisted of various flavonoids with epicatechin, catechin, and quercetin that may be responsible for the effects measured. In conclusion, improvements were seen in rats fed MCL in terms of weight management, endurance capacity, energy metabolism, and muscle injury parameters. PRACTICAL APPLICATIONS: Results of the study revealed that Morinda citrifolia leaf has great potential to be used as functional ingredient in the development of designer food/drink as ergogenic aid for both obese and non-obese individuals. Morinda citrifolia leaf could help in the weight management of obese people and enhance endurance capacity and energy metabolism in active individuals.
The inactivation of antinutritional factors, protease inhibitors within winged bean protein was induced by two respective method treatments. The physical method based on steam vapor that was conducted using an autoclave and chemical method consisting on pH-gradients of buffer solutions prepared at respective acidic pH, neutral pH and alkaline pH ranges. The activity of remaining protease inhibitors of bowman birk inhibitor (BBI), and kunitz-trypsin inhibitor (KTI) after and before treatments was enzymatically confirmed using relevant antagonistic trypsin and combined trypsin-α-chymotrypsin digests. The resulting molecular assembly indicating an interval molecular relaxation range of °0.16 < °DA < °0.2 corresponding to reconformation in protein units with volume-mass changes of -2.17 < ∂v' < +2.17 and with denaturation/unfolding efficiency based on heat capacity ΔCp of 36.36 < DE/UF% < 54.67. These structural changes had a great benefit in determining and producing functional protein hydrolysates.
This study determined the favourable fermentation conditions for the production of antifungal peptides from kenaf seeds and their effectiveness in extending the shelf-life of tomato puree. The optimum fermentation conditions for the maximum activity of the antifungal peptides were 8.4% (w/v), 7 days and 3.7% for substrate/water ratio, fermentation time and glucose concentration, respectively. Eight cationic peptides of low molecular weight ranging from 840 to 1876 Da were identified in kenaf seed peptides mixture (KSPM). The minimum inhibitory concentration and minimum fungicidal concentration of KSPM against Fusarium sp. were 0.18 mg/mL and 0.70 mg/mL, respectively, while those for Aspergillus niger were 1.41 mg/mL and 2.81 mg/mL respectively. KSPM exhibited a fungicidal effect and a prolonged lag phase, with increased fungal membrane permeability as the concentration of KSPM increased, as evidenced by the release of intracellular constituents. The treatment of tomato puree with 1000 mg/kg KSPM delayed fungal growth for up to 14 and 23 days at 25 °C and 4 °C respectively, significantly reducing Aspergillus niger and Fusarium sp. counts. In conclusion, kenaf seed peptides prepared by lacto-fermentation possess antifungal activity, hence can be applied as natural bio preservatives to extend the shelf-life of food products such as tomato puree.
Ambient-storage-friendly, ready-to-eat (RTE) meat products are convenient in emergencies, such as earthquakes, flash floods and the current global Covid-19 lockdown. However, given the processing and long storage time of such food products, the lipid and protein components may be more susceptible to oxidation. Chicken serunding is a low-moisture, high-lipid, high-protein, RTE product that is prone to lipid oxidation and protein co-oxidation, causing product quality deterioration. The present study assessed the effects of storage temperature (25, 40, 60 °C), antioxidant (butylated hydroxyanisole, BHA), and multilayer packaging materials [metallised polyethene terephthalate (MPET) and aluminium] on the lipid oxidation and protein co-oxidation of chicken serunding during six months of storage. All lipid and protein markers elevated with increasing temperature (25 < 40 < 60 °C), indicating that storage of low-moisture meat at high temperature is not feasible. BHA was effective against lipid oxidation, as indicated by the significantly lower (p <0.05) extracted lipid content and delayed formation of malondialdehyde, a secondary lipid oxidation product. However, BHA is not effective against protein co-oxidation, as shown by the insignificant (p >0.05) effect on preventing tryptophan loss, protein carbonyl formation and Schiff base accumulation. MPET packaging with a superior light and oxygen barrier provided significant protection (p <0.05) compared to aluminium. In conclusion, low temperature (25 °C) storage of low-moisture, high-lipid, high-protein, cooked meat systems in MPET packaging is recommended for long-term storage to delay the progression of lipid oxidation and protein co-oxidation.
Fructooligosaccharides can be produced by direct enzymatic conversion from sucrose-rich sugarcane syrup (SS) consisting of 58.93% sucrose yielding 21.28 g FOS/100 g sucrose. This study evaluated the prebiotic effect of unpurified/purified SS containing FOS for the modulation of the human intestinal microbial composition and short-chain fatty acid production. The unpurified and purified FOS substrates, which were a mixture of 1-kestose, nystose and 1F-fructosylnystose, were supplemented into human faecal culture using a pH-controlled batch fermentation system and significantly increased the Bifidobacterium counts after 5 h fermentation, while Bacteroides/Prevotella counts were highest throughout 24 h fermentation. Meanwhile, Lactobacillus/Enterococcus exhibited a slight increase after 5 h fermentation before reaching a plateau afterwards. The steady Bacteroides/Prevotella growth and increased Bifidobacterium population promoted an increase in the production of short-chain fatty acids acetate (58 ± 2.70 mM), propionate (9.19 ± 5.94 mM) and butyrate (7.15 ± 2.28 mM). These results provide evidence that representative gut microbiota could utilise the enzymatically synthesised FOS to generate short-chain fatty acids as metabolites in pH-controlled conditions, thus FOS from SS are a potential prebiotic ingredient for foods and health drinks.
The present work was aimed at investigating hydroethanolic leaf extracts of Cassia fistula for their antioxidant and pancreatic lipase (PL) enzyme inhibitory properties. The most active extract was selected to profile the phytoconstituents by UHPLC-QTOF-MS/MS technique. Among the tested extracts, the 80% hydroethanolic extract exhibited the maximum levels of total phenolic and flavonoid contents (TPC and TFC) with a contribution of 201.3 ± 2.6 mg of gallic acid equivalent per gram of extract (GAE/g extract), and 116.3 ± 2.4 mg of rutin equivalent per gram of extract (RE/g extract), respectively. The same extract also showed promising 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and PL inhibitory activity with an IC50 (half maximal inhibitory concentration) of 30.5 ± 2.8 µg/mL and 17.31 ± 1.18 μg/mL, respectively. The phytochemical profiling of 80% hydroethanolic extract confirmed the presence of 23 metabolites of immense medicinal significance. Docking studies were conducted to investigate the potential interactions of compounds identified in the study. The docking study-based binding energy data and the interaction scheme both revealed the possible role of the identified compounds towards PL inhibitor. Moreover, energies of frontier molecular orbitals (FMOs), ionization potentials (IP), electron affinities (EA) and molecular electrostatic potentials (MEP) were also explored. The findings of the current work suggest that C. fistula is a promising natural source of antioxidant and antiobesity agents, which may be exploited to add pharmacological functionalities to food.
Therapeutic peptides derived proteins with alpha-reconformation states like antibody shape have shown potential effects in combating terrible diseases linked with earlier signs of angiogensis, mutagenesis and transgenesis. Alpha reconformation in material design refers to the folding of the peptide chains and their transitions under reversible chemical bonds of disulfide chemical bridges and further non-covalence lesions. Thus, the rational design of signal peptides into alpha-helix is intended in increasing the defending effects of peptides into cores like adjuvant antibiotic and/or vaccines. Thereby, the signal peptides are able in displaying multiple eradicating regions by changing crystal-depositions and deviation angles. These types of molecular structures could have multiple advantages in tracing disease syndromes and impurities by increasing the host defense against the fates of pathogens and viruses, eventually leading to the loss in signaling by increasing peptide susceptibility levels to folding and unfolding and therefore, formation of transgenic peptide models. Alpha reconformation peptides is aimed in triggering as well as other regulatory functions such as remodulating metabolic chain disorders of lipolysis and glucolysis by increasing the insulin and leptin resistance for best lipid storages and lipoprotein density distributions.
Mung bean protein isolate was texturized at different feed moisture contents (30.0, 49.3, and 60.0%) at a constant temperature (144.57 °C) to evaluate the changes in protein profile, solubility, thermal, structural (at secondary and tertiary levels) and rheological properties. SDS-PAGE, surface hydrophobicity, circular dichroism, FTIR spectroscopy, and fluorescence analyses revealed protein unfolding, aggregation, and structural rearrangement as a function of feed moisture content. Extrusion at 49.3% feed moisture produced texturized mung bean protein (TMBP) with favourable partial denaturation, the formation of small aggregates, improved solubility, and digestibility with strong gel forming behaviour, whereas 30.0 and 60.0% moisture content resulted in complete protein denaturation, the undesirable formation of large aggregates and weak gels. In conclusion, protein denaturation and formation of aggregates can be controlled by manipulating feed moisture content during extrusion, with 49.3% feed moisture prompting favourable partial denaturation to produce TMBP with desirable qualities for use as a vegetarian-based meat extender.
This study aimed to enhance natural gamma aminobutyric acid (GABA) production in yoghurt by the addition of simple sugars and commercial prebiotics without the need for pyridoxal 5'-phosphate (PLP) cofactor. The simple sugars induced more GABA production (42.83-58.56 mg/100 g) compared to the prebiotics (34.19-40.51 mg/100 g), with glucose promoting the most GABA production in yoghurt (58.56 mg/100 g) surpassing the control sample with added PLP (48.01 mg/100 g). The yoghurt prepared with glucose also had the highest probiotic count (9.31 log CFU/g). Simulated gastrointestinal digestion of this GABA-rich yoghurt showed a non-significant reduction in GABA content and probiotic viability, demonstrating the resistance towards a highly acidic environment (pH 1.2). Refrigerated storage up to 28 days improved GABA production (83.65 mg/100 g) compared to fresh GABA-rich yoghurt prepared on day 1. In conclusion, the addition of glucose successfully mitigates the over-use of glutamate and omits the use of PLP for increased production of GABA in yoghurt, offering an economical approach to produce a probiotic-rich dairy food with potential anti-hypertensive effects.
Multiple infections of several bacterial species are often observed under natural farm conditions. The infections would cause a much more significant loss compared to a single infectious agent. Vaccination is an essential strategy to prevent diseases in aquaculture, and oral vaccination has been proposed as a promising technique since it requires no handling of the fish and is easy to perform. This research attempts to develop and evaluate a potential feed-based polyvalent vaccine that can be used to treat multiple infections by Vibrios spp., Streptococcus agalactiae, and Aeromonas hydrophila, simultaneously. The oral polyvalent vaccine was prepared by mixing formalin-killed vaccine of V. harveyi, S. agalactiae, and A. hydrophila strains with commercial feed pellet, and palm oil as an adjuvant was added to improve their antigenicity. Thereafter, a vaccinated feed pellet was tested for feed quality analysis in terms of feed stability in water, proximate nutrient analysis, and palatability, safety, and growth performance using Asian seabass, Lates calcarifer as a fish host model. For immune response analysis, a total of 300 Asian seabass juveniles (15.8 ± 2.6 g) were divided into two groups in triplicate. Fish of group 1 were not vaccinated, while group 2 was vaccinated with the feed-based polyvalent vaccine. Vaccinations were carried out on days 0 and 14 with oral administration of the feed containing the bacterin at 5% body weight. Samples of serum for antibody and lysozyme study and the spleen and gut for gene expression analysis were collected at 7-day intervals for 6 weeks. Its efficacy in protecting fish was evaluated in aquarium challenge. Following vaccination by the polyvalent feed-based vaccine, IgM antibody levels showed a significant (p < 0.05) increase in serum against Vibrio harveyi, Aeromonas hydrophila, and Streptococcus agalactiae and reached the peak at week 3, 5, and 6, respectively. The high-stimulated antibody in the serum remained significantly higher than the control (p < 0.05) at the end of the 6 weeks vaccination trial. Not only that, but the serum lysozyme level was also increased significantly at week 4 (p < 0.05) as compared to the control treatment. The immune-related gene, dendritic cells, C3, Chemokine ligand 4 (CCL4), and major histocompatibility complex class I (MHC I) showed significantly higher expression (p < 0.05) after the fish were vaccinated with the oral vaccine. In the aquarium challenge, the vaccine provided a relative percentage survival of 75 ± 7.1%, 80 ± 0.0%, and 80 ± 0.0% after challenge with V. harveyi, A. hydrophila, and S. agalactiae, respectively. Combining our results demonstrate that the feed-based polyvalent vaccine could elicit significant innate and adaptive immunological responses, and this offers an opportunity for a comprehensive immunization against vibriosis, streptococcosis, and motile aeromonad septicemia in Asian seabass, Lates calcarifer. Nevertheless, this newly developed feed-based polyvalent vaccination can be a promising technique for effective and large-scale fish immunization in the aquaculture industry shortly.
The study was conducted to determine anti-tyrosinase and antioxidant activities of the extracted collagen hydrolysate (CH) derived from Malaysian jellyfish, Rhopilema hispidum. Collagen was extracted using 1:1 (w:v) 0.1 M NaOH solution at temperature 25 °C for 48 hr followed by treatment of 1:2 (w:v) distilled water for another 24 hr and freeze-dried. The extracted collagen was hydrolyzed using papain at optimum temperature, pH and enzyme/substrate ratio [E/S] of 60 °C, 7.0 and 1:50, respectively. CH was found to exhibit tyrosinase inhibitory activity, DPPH radical scavenging and metal ion-chelating assays up to 64, 28, and 83%, respectively, after 8 hr of hydrolysis process. The molecular weight of CH was found <10 kDa consisting of mainly Gly (19.219%), Glu (10.428%), and Arg (8.848%). The UV-visible spectrum analysis showed a major and minor peak at 218 and 276 nm, accordingly. The FTIR spectroscopy confirmed the amide groups in CH. The SEM images demonstrated spongy and porous structure of CH. In the cytotoxicity study, CH has no cytotoxicity against mouse embryonic 3T3 fibroblast cell line with IC50 value >500 µg/ml. Results revealed that the CH generated from this study has a potential to be developed as active ingredient in cosmeceutical application.
The current study evaluated the γ-aminobutyric acid (GABA) producing ability from three novel strains of lactic acid bacteria (L. plantarum Taj-Apis362, assigned as UPMC90, UPMC91, and UPMC1065) co-cultured with starter culture in a yogurt. A combination of UPMC90 + UPMC91 with starter culture symbiotically revealed the most prominent GABA-producing effect. Response surface methodology revealed the optimized fermentation conditions at 39.0 °C, 7.25 h, and 11.5 mM glutamate substrate concentration to produce GABA-rich yogurt (29.96 mg/100 g) with desirable pH (3.93) and water-holding capacity (63.06%). At 2% glucose to replace pyridoxal-5-phosphate (PLP), a cofactor typically needed during GABA production, GABA content was further enhanced to 59.00 mg/100 g. In vivo study using this sample revealed a blood pressure-lowering efficacy at 0.1 mg/kg GABA dosage (equivalent to 30 mg/kg GABA-rich yogurt) in spontaneously hypertensive rats. An improved method to produce GABA-rich yogurt has been established, involving shorter fermentation time and lower glutamate concentration than previous work, along with glucose induction that omits the use of costly PLP, fostering the potential of developing a GABA-rich functional dairy product through natural fermentation with desirable product quality and antihypertensive property.