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Objectives: We investigated salivary biomarkers of stress, more specifically, cortisol and alpha-amylase, to evaluate effects of individualized music listening (IML) in people with dementia. Method: Participants were N = 64 nursing home residents with dementia (meanage = 83.53 ± 7.71 years, 68.8% female). Participants were randomly assigned to either listening to their favorite music every other day for a period of six weeks (intervention), or standard care (control). Using the Saliva Children's Swab (SCS), saliva was collected before, after, and 20 min after IML sessions at the beginning and end of the intervention period for the analysis of salivary alpha-amylase and cortisol. Results: Using the SCS was feasible in people with dementia. Nevertheless, there was no effect of IML on salivary stress markers. Discussion: Although using SCS was feasible, active patient engagement is required. Future studies need to corroborate findings in larger samples. Trial registration: German Clinical Trials Register: DRKS00015641, ISRCTN registry: ISRCTN59052178. [ABSTRACT FROM AUTHOR]

Background: Diabetes mellitus is a metabolic disease characterized by prolonged elevated blood glucose levels. It is a common health problem with a high mortality and morbidity to the human race. A number of medicinal plants such as rosemary (Rosmarinus officinalis) have been used for the treatment of diabetes. Most of the anti-diabetic conventional drugs have been found to have some side effects and there is therefore need to explore new sources of anti-diabetic drugs. The aim of the current study was to investigate the possibility of getting anti-diabetic compounds from R. officinalis that can be used as leads for drug discovery. Methodology: R. officinalis leaves were macerated in 50% methanol in dichloromethane and the crude extract fractionated by column chromatography. The obtained fractions were subjected to an in-vitro alpha-amylase inhibition assay. The anti-hyperglycemic potential of the fractions was evaluated in diabetic induced Wistar rats. The most potent fractions were analyzed by gas chromatography-mass spectrophotometry (GC-MS) for identification of the compounds. Results: A total of 21 chromatographic fractions were assembled with different alpha- amylase inhibition activity. Eleven of the fractions had more than 30% alpha-amylase inhibition activity. The ethyl acetate fraction had the highest inhibition potential (LC50 of 2.8 μg/mL). The anti-diabetic assay in rats showed that fractions (F1) and (F4) had highest blood glucose reduction of 44.5 ± 0.4 and 52.8 ± 1.3%, respectively (p < 0.05). GC-MS analysis of fractions F1 and F4 showed the presence of 21 and 23 compounds in F1 and 23, respectively. Conclusion: This study has demonstrated that R. officinalis crude extract fractions obtained from 50% methanol in dichloromethane possesses alpha-amylase inhibitory and anti-hyperglycemic activities as well as secondary metabolites with varying chemical structures. The hexane and hexane/ethyl acetate (8/2) fractions showed most potent alpha-amylase inhibition with high anti-hyperglycemic activity giving hope of a possibility of obtaining lead compounds for new anti-diabetic drugs. [ABSTRACT FROM AUTHOR]

Postprandial hyperglycemia (PPG) exacerbates endothelial dysfunction and impairs vascular function in diabetes as well in healthy people. Though synthetic drugs are available to regulate PPG, the severe gastrointestinal side effects of those medications have prompted the search for alternative treatments. Recently, some phytochemicals captured the attention because of their inhibitory effects on α-amylase to control diabetes. The aim of this study was to investigate and identify potential alpha-amylase inhibitors in C. indica and W. coagulans. This study also aims to understand one of the possible mechanisms of action of plants for their anti-diabetic activity. A total of 36 phytochemical ligands were subjected for protein–ligand docking analysis. Among the phytochemicals, Taraxerol and Epoxywithanolide-I demonstrated significant binding free energy of − 10.2 kcal/mol and − 11.9 kcal/mol respectively, which was higher than the reference acarbose with − 8.6 kcal/mol. These molecules were subjected for molecular dynamics simulation (MDS) analysis with alpha-amylase protein for a duration of 150 ns. Among the three complexes, Taraxerol and Epoxywithanolide-I complexes demonstrates strong potential as inhibitors of the target protein. MDS results were analyzed via root mean square deviation (RMSD), fluctuation of residues, potential energy, radii of gyration and solvent access surface area analysis. Taraxerol demonstrated a significantly low potential energy of − 1,924,605.25 kJ/mol, and Epoxywithanolide-I demonstrated − 1,964,113.3 kJ/mol of potential energy. RMSD plot shows that Epoxywithanolide-I has much higher stability than the other MDS complexes. Drugability and toxicity studies show that the test ligands are demonstrating strong potential as drug like molecules. The results of the study conclude that, Taraxerol of C. indica and Epoxywithanolide-I of W. coagulans are strong inhibitors of alpha-amylase enzyme and that, this is one of the possible mechanisms of action of the plants for their reported anti-diabetic activities. Further in-vitro analysis is in demand to prove the observed results. [ABSTRACT FROM AUTHOR]

According to the arousal model of vigilance, the locus coeruleus‐norepinephrine (LC‐NE) system modulates sustained attention over long periods by regulating physiological arousal. Recent research has proposed that transcutaneous auricular vagus nerve stimulation (taVNS) modulates indirect physiological markers of LC‐NE activity, although its effects on vigilance have not yet been examined. Aiming to develop a safe and noninvasive procedure to prevent vigilance failures in prolonged tasks, the present study examined whether taVNS can mitigate vigilance loss while modulating indirect markers of LC‐NE activity. Following a preregistered protocol (https://osf.io/tu2xy/), 50 participants completed three repeated sessions in a randomized order, in which either active taVNS at individualized intensity set by participant, active taVNS set at 0.5 mA for all participants, or sham taVNS, was delivered while performing an attentional and vigilance task (i.e., ANTI‐Vea). Changes in salivary alpha‐amylase and cortisol concentrations were measured as markers of LC‐NE activity. Self‐reports of feelings associated with stimulation and guessing rate of active/sham conditions supported the efficacy of the single‐blind procedure. Contrary to our predictions, the observed vigilance decrement was not modulated by active taVNS. Pairwise comparisons showed a mitigation by active taVNS on cortisol reduction across time. Interestingly, Spearman's correlational analyses showed some interindividual effects of taVNS on indirect markers of LC‐NE, evidenced by positive associations between changes in salivary alpha‐amylase and cortisol in active but not sham taVNS. We highlight the relevance of replicating and extending the present outcomes, investigating further parameters of stimulation and its effects on other indirect markers of LC‐NE activity. [ABSTRACT FROM AUTHOR]

Many studies confirm that substances of natural origin have a pronounced affinity for type 2 diabetes mellitus (T2DM) therapeutic targets. At the moment, there is growing interest in bioactive peptides, phytochemicals, and drugs from other natural sources as highly effective, safe and promising antidiabetic agents. Natural sources are a promising resource for regulating several pathological pathways in T2DM. The review describes ways to mitigate insulin resistance and tissue sensitivity to glucose through PTP1β (protein tyrosine phosphatase 1β), GLP-1R (glucagon-like peptide receptor), DPP-4 (dipeptidyl peptidase-4), AMPK (adenosine monophosphate activated protein kinase), MAPK (mitogen-activated protein kinase). The review includes ways to regulate obesity and oxidative stress development through CCN3 (nephroblastoma overexpressed gene), PPAR-γ (peroxisome proliferator-activated receptor γ), Nrf2 (nuclear factor erythroid-related factor 2), FFAR (free fatty acid receptors), 11β-HSD1 (11β-hydroxysteroid dehydrogenase). Moreover, the review details the regulation of hyperglycemia through alpha-amylase inhibitors, as well as regulation of glucose metabolism through GFAT (glutamine fructose6-phosphate aminotransferase), FOXO1 (forkhead box protein O1), GLUT4 (glucose transporter type 4), PGC-1α (receptor gamma coactivator 1α activating peroxisome proliferator). The review examines the use of natural sources, from which low-molecular-weight and peptide compounds are used as T2DM targets modulators. [ABSTRACT FROM AUTHOR]

Immobilization represents one of the strategies to tackle the challenges related to the cost of production, storage stability, and property improvement of biocatalysts. In this study, the Bacillus cereus sp H1 α-amylase was immobilized using physical adsorption onto CaCO3 powder and entrapment in calcium alginate beads. Using experimental design, optimized yields of 80.77% and 97.42% were recorded for the adsorbed and encapsulated amylase, respectively. The surface-modified CaCO3 and Ca-alginate were characterized and confirmed by FTIR and SEM. After immobilization, an upgrade in pH and thermal stabilities was observed with the CaCO3-immobilized amylase compared to the free counterpart and Ca-alginate form. 65% or 52.45% of the activity was conserved after a 1-h incubation at pH 6.5–7.5 or 65°C, respectively. The Ca-alginate-immobilized amylase maintained over 60% of its activity in the presence of 5 mM Cu2+ and Mn2+, whereas the soluble enzyme's activity decreased by 70%. Both immobilized biocatalysts maintained a minimum of 60% activity opposed to surfactants and inhibitors. Nevertheless, the Ca-alginate amylase demonstrated higher resistance to the commercial detergents, preserving at least 88% of its activity. Meanwhile, it retained over than 80% of activity after 35 days of storage at 4°C. The highest stability was observed after 2 reuses, with 55.11% residual activity when using CaCO3 as support. Furthermore, the obtained immobilized amylases were shown to be highly effective in removing starch stains from cotton fabrics, making them potent candidates for developing green and eco-friendly processes, especially for the textile industry. [ABSTRACT FROM AUTHOR]

Rowing races require extreme physical and psychological effort from every athlete. This study aimed to determine the dynamics of the salivary cortisol and alpha-amylase, as well as blood lactate throughout the specific load represented by a 6 km rowing race, conducted on a rowing ergometer. The sample consisted of 11 junior and senior rowers from HVK Gusar in Split (n = 11) who actively competed at club and international levels. Variables consisted of three repeated oral samples of the hormone cortisol and the enzyme alpha-amylase determined in saliva and three repeated blood lactate samples. Potential differences in the levels of the studied variables at different time points were determined using a repeated-measures ANOVA test. The results showed different dynamics of hormonal (cortisol) and metabolic (alpha-amylase and lactates) variables. All variables experienced a significant post-race increase, while other changes were not significant. The results highlighted that high-intensity rowing causes an increase in the body's cortisol, alpha-amylase, and lactate levels. This should be implemented in rowing training to find the right balance between high and low-intensity rowing, enabling athletes' progression while reducing the risk of overtraining. [ABSTRACT FROM AUTHOR]

Alpha-amylase (AMY) plays a significant role in regulating the growth, development, and postharvest quality formation in plants. Nevertheless, little is known about the genome-wide features, expression patterns, subcellular localization, and functional regulation of AMY genes (MaAMYs) in the common starchy banana (Musa acuminata). Twelve MaAMY proteins from the banana genome database were clustered into two groups and contained a conserved catalytic domain. These MaAMYs formed collinear pairs with the AMYs of maize and rice. Three tandem gene pairs were found within the MaAMYs and are indicative of putative gene duplication events. Cis-acting elements of the MaAMY promoters were found to be involved in phytohormone, development, and stress responses. Furthermore, MaAMY02, 08, 09, and 11 were actively expressed during fruit development and ripening. Specifically, MaAMY11 showed the highest expression level at the middle and later stages of banana ripening. Subcellular localization showed that MaAMY02 and 11 were predominately found in the chloroplast, whereas MaAMY08 and 09 were primarily localized in the cytoplasm. Notably, transient attenuation of MaAMY11 expression resulted in an obvious increase in the starch content of banana fruit, while a significant decrease in starch content was confirmed through the transient overexpression of MaAMY11. Together, these results reveal new insights into the structure, evolution, and expression patterns of the MaAMY family, affirming the functional role of MaAMY11 in the starch degradation of banana fruit. [ABSTRACT FROM AUTHOR]

This study presents the phytochemical composition, antioxidant (hydrogen atom and single-atom transfer mechanisms), and digestive enzyme inhibitory (alpha-amylase, alpha-glucosidase, and pancreatic lipase) activities of ethanol-based extractions and traditional preparations (infusion) of the leaves of wild-edible Allium species (A. kharputense, A. affine, A. shirnakiense, and A. akaka) from the highlands of Eastern Anatolia. Among the eight extracts analyzed, ethanol extractions of the A. kharputense and A. akaka leaves exhibited better biotherapeutic activities and had the highest bioactive content. The dominant bioactive profile was composed of mainly allicin and phenolic compounds (chlorogenic acid, hesperidin, rutin, isoquercitrin, and quercetin) with small amounts of fatty acids. These data were similar to the biological activities and chemical composition of common Allium species and suggest the utilization of the extracts of wild-edible Allium species in the development of Allium-based biotherapeutics or nutraceuticals. [ABSTRACT FROM AUTHOR]

Background/Aims: The objectives of our study were to determine salivary α-amylase activity (stress biomarker) and its association with psychological status and quality of life (QoL), disease duration and intensity of symptoms (pain/burning) in patients with OLP. Methods: A total of 50 subjects participated in this case-control study: 30 patients with oral lichen planus (OLP); 20 control subjects. Unstimulated whole saliva (UWS) was collected between 9 and 10 am to avoid diurnal fluctuations. Psychological status was assessed using the Croatian validated version of the original Depression, Anxiety and Stress Scale (DASS-21). The impact of oral health on QoL was assessed using the Croatian version of the Oral Health Impact Profile Questionnaire (OHIP-CRO14). Results: There was no statistically significant difference in salivary α-amylase activity between patients with OLP (N=30) and control subjects (N=20) (133813.3 vs. 166815.5 U/L, p=0.314; t-test). Depression, anxiety and stress showed no statistically significant difference between patients with OLP and control subjects (p=0.076, p=0.111, p=0.209; t-test). The patients with OLP had statistically significantly poorer QoL (total) compared to control subjects (p=0.004, t-test). There was a moderate positive correlation between symptom intensity (pain/burning) and poor QoL (total) (r=0.584, p<0.001), the OHIP-CRO14 dimension "physical pain" (r=0.661, p<0.001), "psychological impossibility" (r=0.555, p<0.01), "handicap" (r=0.546, p<0.01). Conclusion: Although salivary α-amylase showed no statistically significant difference between patients with OLP and control subjects, the patients with OLP had poorer psychological status (three times higher scores for depression and two times higher scores for anxiety) and poorer QoL compared to the control subjects. Recognising and treating mental disorders in patients with OLP is important in order to break the „vicious circle” and achieve a better QoL in these patients. [ABSTRACT FROM AUTHOR]

Non-invasive transcutaneous auricular vagus nerve stimulation (taVNS) has attracted increasing interest as a neurostimulation tool with potential applications in modulating cognitive processes such as attention and memory, possibly through the modulation of the locus–coeruleus noradrenaline system. Studies examining the P300 brain-related component as a correlate of noradrenergic activity, however, have yielded inconsistent findings, possibly due to differences in stimulation parameters, thus necessitating further investigation. In this event-related potential study involving 61 participants, therefore, we examined how changes in taVNS parameters, specifically stimulation type (interval vs. continuous stimulation) and duration, influence P300 amplitudes during a visual novelty oddball task. Although no effects of stimulation were found over the whole cluster and time window of the P300, cluster-based permutation tests revealed a distinct impact of taVNS on the P300 response for a small electrode cluster, characterized by larger amplitudes observed for easy targets (i.e., stimuli that are easily discernible from standards) following taVNS compared to sham stimulation. Notably, our findings suggested that the type of stimulation significantly modulated taVNS effects on the P300, with continuous stimulation showing larger P300 differences (taVNS vs. sham) for hard targets and standards compared to interval stimulation. We observed no interaction effects of stimulation duration on the target-related P300. While our findings align with previous research, further investigation is warranted to fully elucidate the influence of taVNS on the P300 component and its potential utility as a reliable marker for neuromodulation in this field. [ABSTRACT FROM AUTHOR]

The Asian region of earth has a rich agriculture system that provides extensive opportunities to boost pharmaceutical and nutritional research to address the use of food crops for health benefits and potential clinical applications. The traditionally cropped green leaf vegetable indigenously known as Bathu and botanically known as Chenopodium album (C. album) is traditionally used as a sedative, blood purifier, hepatoprotectant, diuretic, and antiscorbutic laxative. In this study, we investigated the anti-infection potential, anti-diabetic potential, and mineral composition of indigenously cultivated C. album plant extracts. Methanol and n-hexane solvents were used to extract phytochemicals at different extraction conditions. The maximum yield of 12.72 ± 0.36 g/100 g extract was obtained in methanol with 200 rpm shaking, 200 mL solvent, and an 8 h extraction period. Under the same conditions, n-hexane gave 2.09 ± 0.29 g/100 g extract. Good alpha-amylase inhibition efficiency was shown by the n-hexane extracts, while the methanol extracts showed good urease inhibition potential. The H6 extract had the lowest IC50 (8.16 ± 0.2 ug/mL) as compared to the standard acarbose (9.27 ± 0.6 ug/mL). Similarly, the M6 extract revealed a significant urease inhibitory potential, i.e., IC50 of 18.77 ± 0.6 ug/mL, which was close to the standard thiourea (IC50: 19.09 ± 0.7 ug/mL). Regarding the antibacterial study, the M6 extract showed 16.55 ± 0.57 mm ZOI against E. coli and 15.54 ± 0.55 mm in the case of S. aureus, as compared to the standard ciprofloxacin, which showed 26.08 ± 0.73 mm, and penicillin, which showed 21.12 ± 0.81 mm ZOI. Mineral profiling was investigated by ICP-OES, which showed significant amounts of Mg and Fe in all extracts. Our findings tend to show that systematic harvesting and utilization of this vegetable crop could be recommended as an alternative nutritional therapy in the management of internal infections and diabetes. [ABSTRACT FROM AUTHOR]

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of the alpha‐amylase (Ronozyme® HiStarch CT/L) produced with a genetically modified strain of Bacillus licheniformis (DSM 34315) as a zootechnical feed additive for chickens for fattening, turkeys for fattening and minor growing poultry species. The additive is available in two forms, a coated thermotolerant granulate formulation and a liquid formulation. The production strain and its DNA were not detected in an intermediate concentrated product representative of the final formulations. The Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) concluded that the final product did not trigger safety concerns with regard to the genetic modification. The Panel concluded that Ronozyme® HiStarch CT/L is safe for chickens for fattening, turkeys for fattening and minor growing poultry species at the recommended inclusion level of 80 KNU/kg complete feed. The use of Ronozyme® HiStarch CT/L in animal nutrition under the proposed conditions of use is safe for the consumers and the environment. The additive in any form was shown to be non‐irritant to the skin and the solid form was shown to be non‐irritant to the eyes. No conclusions could be drawn on the potential of the liquid form to be irritant to the eyes or on the potential of both forms of the additive to be dermal sensitisers due to lack of data. Owing to the proteinaceous nature of the active substance, the additive in either form was considered to be a respiratory sensitiser and any exposure by inhalation is considered a risk. In the absence of data, the Panel was not in the position to conclude on the efficacy of Ronozyme® HiStarch CT/L for chickens for fattening, turkeys for fattening and minor growing poultry species. [ABSTRACT FROM AUTHOR]

In this study, propolis referred to as of Türkiye Anatolian propolis was utilized. This work was aim to prepare a 70% ethanolic extract of propolis and subsequently determine certain biochemical parameters, total phenolic content (TPC), total flavonoid content (TFC), phenolic and flavonoid composition, inhibitory effects on certain enzymes [acetylcholinesterase (AChE), urease, and alpha-amylase], along with assessments of antiviral activity against Vero cells, lung cancer cells (A549), and breast cancer cells (MDA-MB-231). The richness of phenolic content in Anatolian propolis clearly indicates a high inhibitory effect on these enzymes. Additionally, it was observed to exert significant effects on cancer cells and demonstrated effective antiviral activity. It can be contemplated that the Anatolian propolis might serve as a potential source for novel drugs, and the isolation of its compounds could offer opportunities for utilization in traditional medicine. [ABSTRACT FROM AUTHOR]

This study emphasized the alpha-amylase gene from Bacillus licheniformis as this species has been witnessed as the most prolific bio-factories for the alpha amylase production on industrial scales with particular reference to its wide commercial applicability. In the present study, a gene encoding alpha-amylase from an indigenously isolated B. licheniformis 208 strain was PCR amplified and sequenced. The obtained 1313 bp long nucleotide sequence (GenBank KM27230) exhibited the closest match with its homologous genes in B. licheniformis ATCC 9945A and ATCC 14580 strains. Gene translation retrieved 437 amino acid long enzyme sequence and exhibits a molecular weight of 50231.8 KDa. Other computed parameters include isoelectric point (5.95), aliphatic index (67.4), instability index (27.45), and GRAVY (-0.627) values. The structure of alphaamylase 208 was found to exhibit a 17 residues long active site, a conserved catalytic site with 3 residues, and a ligand-binding site comprised of 3 residues. Secondary structure configuration in the alpha-amylase 208 was predominated with a-helix and random coils followed by extended strands. The number of ß-turns was found comparatively less. ITASSER assigned E.C number to the BLA 208 sequence was 3.2.1.1 with extracellular origin. Deduced tertiary structure of BLA 208 showed the 7 residues long active site comprising of histidine, arginine, aspartic acid, glutamic acid, tryptophan, histidine, and aspartic acid at position 122, 246, 248, 278, 280, 344, 345 respectively. Na, K, and Ca were observed as the ligand-binding sites existing as a metal triad. Natural selection analysis explains that the alpha-amylase gene in B. licheniformis species is under purifying or negative selection i.e. it stabilizes itself by removing the deleterious mutation. Recombination event analysis was done using GARD (Genetic algorithm for recombination detection). Although, it rather inferred the presence of two break points which were found insignificant when validated via Kishino-Hasegawa test which demonstrates topological incongruence. [ABSTRACT FROM AUTHOR]

Barnyard millet (Echinochloa frumantacea L.) is a traditional South Indian dietary staple known for its short grain ripening period of 45 days. Various processing methods are employed to enhance its nutrient quality, aid digestion, and improve bioavailability by reducing anti-nutrients. This study aimed to compare the nutritional properties of barnyard millet processed through different methods, including whole flour, de-husked flour, and roasted flour. Samples were collected from different regions in Tamil Nadu, namely Chennai, Kovilpatti, and Virudhunagar. Subsequently, they underwent maceration extraction using methanol to evaluate their nutritional, antioxidant, and anti-nutritional properties. Phytochemical analysis revealed elevated levels of selected compounds in all samples, with processing methods resulting in a reduction of anti-nutrient content. Notably, the roasted sample from Chennai exhibited significant antioxidant activity, while de-husked flour from Chennai and Kovilpatti regions demonstrated inhibition of in vitro anti-obesity activity. RP-HPLC analysis identified major metabolites, and molecular docking against oxidoreductase proteins highlighted the efficacy of Chlorogenic acid and Gallic acid. These findings suggest the potential for developing novel therapeutic drugs for diabetes and anti-cancer health benefits associated with specific barnyard millet processing methods. [ABSTRACT FROM AUTHOR]

Juniperus oxycedrus is prevalent in the arid and semi-arid regions of the Mediterranean and Near East and, is traditionally employed for both culinary and medicinal applications. This research evaluates the potential health benefits of ethyl acetate (EAJO) and n-butanol (NBJO) fractions derived from leaves. Quantitative analysis revealed enhanced concentrations of phenolic and flavonoid compounds in the EAJO fraction at 252.56 ± 3.91 mg GAE/g DW and 47.70 ± 2.21 mg QE/g DW, respectively. UPLC-ESI-MS/MS identified a rich array of twelve phenolic acids and flavonoids, with rutin and chrysin identified as the predominant compounds in both fractions. Comprehensive antioxidant assessments through seven distinct assays demonstrated strong properties, with most activities displaying an IC50 values under 30 μg/mL. Additionally, in vitro anti-inflammatory activity showed that the EAJO and NBJO fractions effectively inhibited albumin denaturation at different concentrations, indicating a moderate effect compared to the standard drug diclofenac sodium. EAJO fraction surpassed NBJO in inhibiting key enzymes associated with Alzheimer's disease, diabetes, hyperpigmentation, dermatological disorders, and select bacterial infections, evidenced by IC50 values for acetylcholinesterase, alpha-amylase, tyrosinase, and urease with IC50 values of 14.60 ± 0.98 μg/mL, 639.48 ± 7.43 μg/mL, 206.44 ± 18.48 μg/mL, and 245.77 ± 3.75 μg/mL, respectively. Both fractions also displayed promising photoprotective properties, with SPF ratings of 31.71 ± 0.31 for NBJO and 22.62 ± 1.52 for EAJO. This study highlights the significant potential of Juniperus oxycedrus in the development of pharmaceutical, cosmetic, and nutritional products, underlining its substantial bioactive profile with antioxidant, anti-inflammatory, anti-enzymatic, and photoprotective capabilities. [ABSTRACT FROM AUTHOR]

As a rule, an experiment carried out at school or in undergraduate study courses is rather simple and not very informative. However, when the experiments are to be performed using modern methods, they are often abstract and difficult to understand. Here, we describe a quick and simple experiment, namely the enzymatic characterization of ptyalin (human salivary amylase) using a starch degradation assay. With the experimental setup presented here, enzyme parameters, such as pH optimum, temperature optimum, chloride dependence, and sensitivity to certain chemicals can be easily determined. This experiment can serve as a good model for enzyme characterization in general, as modern methods usually follow the same principle: determination of the activity of the enzyme under different conditions. As different alleles occur in humans, a random selection of test subjects will be quite different with regard to ptyalin activities. Therefore, when the students measure their own ptyalin activity, significant differences will emerge, and this will give them an idea of the genetic diversity in human populations. The evaluation has shown that the pupils have gained a solid understanding of the topic through this experiment. [ABSTRACT FROM AUTHOR]

Abstract Background Diabetes mellitus is a metabolic disease characterized by prolonged elevated blood glucose levels. It is a common health problem with a high mortality and morbidity to the human race. A number of medicinal plants such as rosemary (Rosmarinus officinalis) have been used for the treatment of diabetes. Most of the anti-diabetic conventional drugs have been found to have some side effects and there is therefore need to explore new sources of anti-diabetic drugs. The aim of the current study was to investigate the possibility of getting anti-diabetic compounds from R. officinalis that can be used as leads for drug discovery. Methodology R. officinalis leaves were macerated in 50% methanol in dichloromethane and the crude extract fractionated by column chromatography. The obtained fractions were subjected to an in-vitro alpha-amylase inhibition assay. The anti-hyperglycemic potential of the fractions was evaluated in diabetic induced Wistar rats. The most potent fractions were analyzed by gas chromatography-mass spectrophotometry (GC-MS) for identification of the compounds. Results A total of 21 chromatographic fractions were assembled with different alpha- amylase inhibition activity. Eleven of the fractions had more than 30% alpha-amylase inhibition activity. The ethyl acetate fraction had the highest inhibition potential (LC50 of 2.8 μg/mL). The anti-diabetic assay in rats showed that fractions (F1) and (F4) had highest blood glucose reduction of 44.5 ± 0.4 and 52.8 ± 1.3%, respectively (p

Abstract Aromatic compounds are known anti-amyloid aggregates. Their effect on amorphous aggregates of proteins is, however, less studied. We chose aromatic amino acids Trp, Tyr, and Phe, as well as another known stabilizer (i.e. Arg), as potential compatible solvents to be tested on Bacillus amyloliquefaciens alpha-amylase (BAA). Among these additives, Phe was the only one to be effective on the thermal inactivation and amorphous aggregation of BAA, while preserving its intrinsic activity. A concentration of 50 mM Phe was used to test its potential in counteracting the deleterious effect of BAA amorphous aggregates in vivo. After 21 days of daily subcutaneous injections of the native enzyme to mice, amorphous aggregates of BAA, as well as aggregates produced in presence of 50 mM Phe, the tissues located at the site of injection were studied histologically. Amorphous aggregates caused an increase in macrophages and lipid droplets. Serum levels of IL6 and TNF-α were also accordingly elevated and indicative of an inflammation state. Aggregates also resulted into increased levels of glucose, triglycerides and cholesterol, as well as liver enzymes SGOT and SGPT. On the other hand, the presence of Phe prevented this exacerbated inflammatory state and the subsequent impairment of biochemical parameters. In conclusion, Phe is an interesting compound for both stabilizing proteins and counteracting the pathological effect of amorphous aggregates.

Plants possess endless structural and chemical diversity, which is peerless with any synthetic library of small biomolecules, inspiring novel drug discovery. Plants are widely applied to encounter global health challenges such as antimicrobial resistance and diabetes. The objective of this work was to evaluate the antibacterial and antidiabetic potency of native plants grown in Cyprus. All plants were sequentially extracted with solvents of increasing polarity, namely hexane, acetone, methanol, and water. First, the phenolic and flavonoid contents of the extracts were assessed. Afterwards, the bacteriostatic and bactericidal potency of plant extracts were tested against a panel of six bacteria using the broth microdilution method, whereas the inhibitory effects on alpha-glucosidase and alpha-amylase enzymes were also determined with the employment of microplate assays. The results highlighted the superiority of Sarcopoterium spinosum as a potential enzyme inhibitor, while a knowledge base was also acquired for the inhibitory potential of all plants. Daucus carota, Ferula communis, and Tordylium.aegyptiacum displayed additionally outstanding bacteriostatic and bactericidal effects on Gram-positive bacteria at concentrations of 250 µg mL−1 and 500 µg mL−1. Overall, the present study describes the antibacterial and inhibitory activity against carbohydrate digestive enzymes of native plants grown in Cyprus delivering the first reports for many plant species.

Diabetes mellitus is a persistent metabolic condition marked by elevated blood glucose levels due to compromised insulin secretion or functionality. The search for natural antidiabetic agents has gained attention due to their potential effectiveness and safety profiles. Sessuvium portulacastrum, a coastal plant, has been traditionally used for various medicinal purposes. This study investigates the antidiabetic potential of Sessuvium portulacastrum aqueous extract by analyzing its inhibitory effects on key enzymes involved in carbohydrate metabolism and exploring its molecular interactions with critical target proteins. The aqueous extract of Sessuvium portulacastrum was prepared and used for in vitro analysis. The reduced activity of the extract against α-amylase and α-glucosidase enzymes, crucial in glucose absorption and postprandial hyperglycemia, was assessed. Molecular docking techniques were employed to explore the potential interactions between active compounds in the extract and diabetes-related proteins, including BAX, GSK3β, and CADH. The study revealed significant inhibition of both alpha-amylase and alpha-glucosidase enzymes by Sessuvium portulacastrum aqueous extract, indicating its potential to reduce glucose absorption and postprandial hyperglycemia. Moreover, the molecular docking analysis demonstrated strong binding interactions between active compounds in the extract and key proteins involved in diabetes-related pathways, namely apoptotic pathways, glycogen synthesis, and cell adhesion. The findings of this study highlight the promising antidiabetic potential of Sessuvium portulacastrum aqueous extract. Upcoming research should get an attention on isolating and characterizing the active compounds responsible for these effects on antidiabetic therapies from natural sources.

Aromatic compounds are known anti-amyloid aggregates. Their effect on amorphous aggregates of proteins is, however, less studied. We chose aromatic amino acids Trp, Tyr, and Phe, as well as another known stabilizer (i.e. Arg), as potential compatible solvents to be tested on Bacillus amyloliquefaciens alpha-amylase (BAA). Among these additives, Phe was the only one to be effective on the thermal inactivation and amorphous aggregation of BAA, while preserving its intrinsic activity. A concentration of 50 mM Phe was used to test its potential in counteracting the deleterious effect of BAA amorphous aggregates in vivo. After 21 days of daily subcutaneous injections of the native enzyme to mice, amorphous aggregates of BAA, as well as aggregates produced in presence of 50 mM Phe, the tissues located at the site of injection were studied histologically. Amorphous aggregates caused an increase in macrophages and lipid droplets. Serum levels of IL6 and TNF-α were also accordingly elevated and indicative of an inflammation state. Aggregates also resulted into increased levels of glucose, triglycerides and cholesterol, as well as liver enzymes SGOT and SGPT. On the other hand, the presence of Phe prevented this exacerbated inflammatory state and the subsequent impairment of biochemical parameters. In conclusion, Phe is an interesting compound for both stabilizing proteins and counteracting the pathological effect of amorphous aggregates. [ABSTRACT FROM AUTHOR]

Diabetes mellitus is one of the fastest-growing metabolic disorders. Worldwide, one out of 10 people suffer from diabetes mellitus. In this manuscript, we establish the antidiabetic potential of previously synthesized pyridazine derivatives using in silico and in vitro experiments. Molecular docking studies of pyridazine derivatives with pullulanase Amy12, isomaltase, and alpha-amylase receptors revealed that Molecules 45, 46, and 29 showed good docking interaction scores. ADMET analysis of the top three docked pyridazine derivatives (45, 46, and 29) showed lesser solubility and poor distribution behavior. Molecules 45, 46, and 29 maintained drug-like properties with good oral bioavailability. Molecules 45 and 29 showed reproductive toxicity without any sign of mutagenicity, tumorigenicity, or other irritancy. Molecule 46 emerged as a nontoxic molecule in all aspects. In vitro alpha-amylase inhibition data of molecules 45, 46, 29, and acarbose showed IC50 values of 8.67, 8.46, 8.83, and 66.65 µg/mL, respectively. The antidiabetic activity of pyridazine derivatives is increased with the presence of phenothiazine coupled with a biphenyl group, indole with a biphenyl group, and phenothiazine with an ethylphenyl group. Establishing these pyridazine derivatives using in vivo experiments could be beneficial in managing diabetes. [ABSTRACT FROM AUTHOR]

Although enchytraeids have gained popularity in scientific research, fundamental questions regarding their feeding ecology and biology remain largely unexplored. This study investigates α-amylases, major digestive enzymes responsible for hydrolyzing starch and similar polysaccharides into sugars, in Enchytraeus albidus. Genetic data related to α-amylases is currently lacking for the family Enchytraeidae but also for the entire Annelida. To detect and identify coding sequences of the expressed α-amylase genes in COI-monohaplotype culture (PL-A strain) of E. albidus , we used classical "gene fishing" and transcriptomic approaches. We also compared coding sequence variants of α-amylase retrieved from transcriptomic data related to freeze-tolerant strains. Our results reveal that E. albidus possesses two distinct α-amylase genes (Amy I and Amy II) that are homologs to earthworm Eisenia fetida Ef-Amy genes. Different strains of E. albidus possess distinctive alleles of α-amylases with unique SNP patterns specific to a particular strain. Unlike Amy II, Amy I seems to be a highly polymorphic and multicopy gene. The domain architecture of the putative Amy proteins was found the same as for classical animal α-amylases with ABC-domains. A characteristic feature of Amy II is the lack of GHGA motif in the flexible loop region, similarly to many insect amylases. We identified " Enchytraeus - Eisenia type" α-amylase homologs in other clitellates and polychaetes, indicating the ancestral origin of Amy I/II proteins in Annelida. This study provides the first insight into the endogenous non-proteolytic digestive enzyme genes in potworms, discusses the evolution of Amy α-amylases in Annelida, and explores phylogenetic implications. [Display omitted] • E.albidus has 2 α-amylase genes (Amy I and II) homologous to E. fetida Ef-Amy genes. • Different E. albidus strains have unique amylase alleles with specific SNPs. • Amy I seems to be a polymorphic and multicopy gene, unlike Amy II. • The loss of GHGA motif in the flexible loop of Amy II is ancestral in Clitellata. • " Enchytraeus - Eisenia type" α-amylase homologs are present in other annelids. [ABSTRACT FROM AUTHOR]

Acarbose is a potent glycosidase inhibitor widely used in the clinical treatment of type 2 diabetes mellitus (T2DM). Various acarbose analogs have been identified while exploring compounds with improved pharmacological properties. In this study, we found that AcbE from Actinoplanes sp. SE50/110 catalyzes the production of acarbose analogs that exhibit significantly improved inhibitory activity towards α-amylase than acarbose. Recombinant AcbE mainly catalyzed the formation of two new compounds, namely acarstatins A and B, using acarbose as substrate. Using high-resolution mass spectrometry, nuclear magnetic resonance, and glycosidase hydrolysis, we elucidated their chemical structures as O-α-D-maltosyl-(1 → 4)-acarbose and O-α-D-maltotriosyl-(1 → 4)-acarbose, respectively. Acarstatins A and B exhibited 1584- and 1478-fold greater inhibitory activity towards human salivary α-amylase than acarbose. Furthermore, both acarstatins A and B exhibited complete resistance to microbiome-derived acarbose kinase 1-mediated phosphorylation and partial resistance to acarbose-preferred glucosidase-mediated hydrolysis. Therefore, acarstatins A and B have great potential as candidate therapeutic agents for T2DM. [ABSTRACT FROM AUTHOR]

High thermostability and catalytic efficiency of α-amylase are required for the industrial applications. To improve catalytic parameters including Michaelis constant, maximum velocity, turnover number, enzyme half-life, and the catalytic efficiency of α-amylase, it was treated alone and/or simultaneously with Ca2+ ions (12.5 and 25 mM) and/or ultrasound (USN) (64.5 W, 25 + 40 kHz). The structural analysis of the enzyme (the enzyme's secondary structure and conformation) was carried out using Fourier transform infrared (FTIR) spectroscopy and circular dichroism (CD) techniques. Moreover, the thermal properties of the enzyme were evaluated using differential scanning calorimetry (DSC). USN + 12.5 mM Ca2+ and 25 mM Ca2+, changed the maximum velocity (+ 6.5% and + 37.4%), turnover number (+ 6.6% and + 38.2%), enzyme half-life (+ 51.6% and + 113.7%), and catalytic efficiency (+ 18.1% and -3.3%) of α-amylase. The energy barriers of thermo-inactivation and inactivation enthalpy of α-amylase exposed to USN + 25 mM Ca2+ were increased by 25% and 5.8%, respectively, without affecting entropy of inactivation, showing how enzyme's thermal stability was improved after the treatments. The increase in midpoint temperature (35%) and α-helix (14%) values as well as the decrease in the β-sheet structure (39%) of the α-amylase after applying USN + Ca2+ (25 mM) confirmed the thermal stability enhancement of α-amylase after the treatment compared to control. [ABSTRACT FROM AUTHOR]

Exploring the natural product as a potential alpha-amylase inhibitor with no adverse side effects is still being concerned in developing diabetes mellitus drugs. This study aims to identify the phytochemical components of porang tuber extract. We also investigated bioactivity and predicted the biological function of the porang tubers bioactive in inhibiting human pancreatic alpha-amylase. The phytochemical components were analyzed by Liquid Chromatography Mass Spectrophotometry (LCMS). We investigated the biological activity of compounds as anti-diabetic agents by the PASS server. The 3D structure of the active compound that was showing anti-diabetic activity was prepared using PyRx software. Docking simulations were analyzed using Hex 8.0 software, while for visualization Discovery Studio Client 4.1 software was used. Our study found the presence of 67 phytochemical constituents in porang tuber extract. The five largest compounds with anti-diabetic activity are glucomannan, mannan, and flavonoids (quercetin, orientin, and hyperoside). PASS analysis showed that glucomannan and mannan had the most potential as anti-diabetic. It was determined by the Pa value of 0.77. The active sites of alpha-amylase (ASP197, GLU233, and ASP300 residues) were bound by glucomannan, mannan, orientin, and hyperoside, which plays an essential role in catalytic activity. Glucomannan showed the strongest interaction with the pancreatic alpha-amylase domain (lowest binding energy -394.1 kcal/mol). This result indicates that porang tubers are very potential as an alpha-amylase inhibitor. Further research is needed to validate this finding. [ABSTRACT FROM AUTHOR]

Plants possess endless structural and chemical diversity, which is peerless with any synthetic library of small biomolecules, inspiring novel drug discovery. Plants are widely applied to encounter global health challenges such as antimicrobial resistance and diabetes. The objective of this work was to evaluate the antibacterial and antidiabetic potency of native plants grown in Cyprus. All plants were sequentially extracted with solvents of increasing polarity, namely hexane, acetone, methanol, and water. First, the phenolic and flavonoid contents of the extracts were assessed. Afterwards, the bacteriostatic and bactericidal potency of plant extracts were tested against a panel of six bacteria using the broth microdilution method, whereas the inhibitory effects on alpha-glucosidase and alpha-amylase enzymes were also determined with the employment of microplate assays. The results highlighted the superiority of Sarcopoterium spinosum as a potential enzyme inhibitor, while a knowledge base was also acquired for the inhibitory potential of all plants. Daucus carota, Ferula communis, and Tordylium.aegyptiacum displayed additionally outstanding bacteriostatic and bactericidal effects on Gram-positive bacteria at concentrations of 250 µg mL−1 and 500 µg mL−1. Overall, the present study describes the antibacterial and inhibitory activity against carbohydrate digestive enzymes of native plants grown in Cyprus delivering the first reports for many plant species. [ABSTRACT FROM AUTHOR]

The present study screened various fungal species for inhibitors of alpha-glucosidase, alpha-amylase, and DPP-4, enzymes that are crucial in carbohydrate metabolism. Ethanolic extracts exhibited superior inhibitory activity compared to water extracts, suggesting their potential as sources of anti-diabetic agents. Further fractionation revealed fomentariol from Fomes fomentarius as a potent inhibitor of alpha-glucosidase and DPP-4, with higher activity against alpha-glucosidase than acarbose. Fomentariol presents a novel avenue for diabetes management, demonstrating the simultaneous inhibition of key enzymes in glucose metabolism. However, comprehensive clinical studies are needed to evaluate its safety and efficacy in humans. [ABSTRACT FROM AUTHOR]

The coordination of assimilation pathways for all the elements that make up cellular components is a vital task for every organism. Integrating the assimilation and use of carbon (C) and nitrogen (N) is of particular importance because of the high cellular abundance of these elements. Starch is one of the most important storage polymers of photosynthetic organisms, and a complex regulatory network ensures that biosynthesis and degradation of starch are coordinated with photosynthetic activity and growth. Here, we analyzed three starch metabolism enzymes of Chlamydomonas reinhardtii that we captured by a cyclic guanosine monophosphate (cGMP) affinity chromatography approach, namely, soluble starch synthase STA3, starch‐branching enzyme SBE1, and α‐amylase AMA2. While none of the recombinant enzymes was directly affected by the presence of cGMP or other nucleotides, suggesting an indirect binding to cGMP, AMA2 activity was stimulated in the presence of L‐glutamine (Gln). This activating effect required the enzyme's N‐terminal aspartate kinase–chorismate mutase–tyrA domain. Gln is the first N assimilation product and not only a central compound for the biosynthesis of N‐containing molecules but also a recognized signaling molecule for the N status. Our observation suggests that AMA2 might be a means to coordinate N and C metabolism at the enzymatic level, increasing the liberation of C skeletons from starch when high Gln levels signal an abundance of assimilated N. [ABSTRACT FROM AUTHOR]

Horse welfare is the product of multiple factors, including behavioral and physiological adjustments to cope with stressful situation regarding environment and housing condition. Collectively, it is supposed that a horse kept in the wild has a lower level of stress than other housing system, and the aim of the present study was to investigate the level of stress in domestic horses reared in the wild and then moved to human controlled housing, through saliva analysis. Twelve clinically healthy Catria (Italian local breed) mares, usually reared in the wild, were moved into collective paddocks for a folkloric event. Saliva samples were obtained before and after the change of housing condition to evaluate stress biomarkers including salivary cortisol, salivary alpha-amylase, and butyrylcholinesterase (BChol). The mares were also scored using the Welfare Aggregation and Guidance (WAG) Tool to highlight the presence of abnormal behaviors. Despite the absence of differences in behavioral scores between wild and paddocks, salivary cortisol and BChol were found to be higher in the wild and lower when mares were moved to paddocks. The highest concentrations in stress biomarkers like salivary cortisol and BChol in the wild was unexpected, but the need for managing hierarchical relationships, and the exposure to feral animals, predators, and weather changes, might explain these findings. The overall results of the present study may provide further knowledge toward stress response in domesticated horses living in the wild moved to human controlled housing system. [ABSTRACT FROM AUTHOR]

[{TiCl3}2(C3N3S3H)] (where C3N3S3H is Trithiocyanuric acid) was designed and synthesized by the reaction of TiCl4 and trithiocyanuric acid in 2:1 M ratio under stirring and refluxing condition using THF solvent. The synthesized Titanium (IV) complex was characterized by various spectroscopic techniques like FT‐IR, NMR, MASS, XRD, UV–visible spectrophotometer and elemental analysis (CHNSO). Moreover, in‐silico docking studies of the ligand and its Ti (IV) complex were carried out by AutoDockTools‐1.5.6 to study interactions of the complex under study with the receptor protein. Afterwards, both ligand and synthesized Ti (IV)complex were screened for the antioxidant and antidiabetic potential by in‐vitro method. The antioxidant potential was investigated by using DPPH (2,2‐diphenyl‐1‐picrylhydrazyl) radical scavenging assay where ligand exhibited moderate activity while Ti (IV) complex presented significant antioxidant activity. Following, alpha‐amylase enzyme inhibition was investigated using iodine‐starch inhibition assay. It was found that the ligand was inactive (showed no α‐amylase inhibition activity) while the Ti (IV)complex was a potent α‐amylase inhibitor. Furthermore, the experimentally obtained results were also complimented by the in‐silico results. [ABSTRACT FROM AUTHOR]

Diabetes mellitus is a persistent metabolic condition marked by elevated blood glucose levels due to compromised insulin secretion or functionality. The search for natural antidiabetic agents has gained attention due to their potential effectiveness and safety profiles. Sessuvium portulacastrum, a coastal plant, has been traditionally used for various medicinal purposes. This study investigates the antidiabetic potential of Sessuvium portulacastrum aqueous extract by analyzing its inhibitory effects on key enzymes involved in carbohydrate metabolism and exploring its molecular interactions with critical target proteins. The aqueous extract of Sessuvium portulacastrum was prepared and used for in vitro analysis. The reduced activity of the extract against a-amylase and a-glucosidase enzymes, crucial in glucose absorption and postprandial hyperglycemia, was assessed. Molecular docking techniques were employed to explore the potential interactions between active compounds in the extract and diabetes-related proteins, including BAX, GSK3ß, and CADH. The study revealed significant inhibition of both alpha-amylase and alpha-glucosidase enzymes by Sessuvium portulacastrum aqueous extract, indicating its potential to reduce glucose absorption and postprandial hyperglycemia. Moreover, the molecular docking analysis demonstrated strong binding interactions between active compounds in the extract and key proteins involved in diabetes-related pathways, namely apoptotic pathways, glycogen synthesis, and cell adhesion. The findings of this study highlight the promising antidiabetic potential of Sessuvium portulacastrum aqueous extract. Upcoming research should get an attention on isolating and characterizing the active compounds responsible for these effects on antidiabetic therapies from natural sources. [ABSTRACT FROM AUTHOR]

In this study, an efficient protocol was developed to obtain multiplicated, healthy sweet basil plantlets with larger leaves and a greener color by culturing them on MS medium containing 1.0 mg/l 2iP. Chitosan and zinc oxide nanoparticles (ZnO NPs) were used as abiotic elicitors to maximize the main secondary metabolites (phenolic and flavonoid compounds and essential oil) in in vitro plantlets of sweet basil. Chitosan with a concentration of 4.0 mg/l recorded the maximum value of caffeic acid (1.189 mg/g DW), and 10 mg/l of ZnO NPs recorded the maximum content of rutin (0.936 mg/g DW). The plantlets cultured with 2.0 mg/l chitosan recorded the maximum percentage of essential oil (90.63%). 8,9-Dihydro-9-formyl cycloisolongifolene was the major compound (72.44%) and was detected in the plantlets treated with 2.0 mg/l chitosan. The inhibitory activity evaluation of α-amylase revealed that the maximum inhibitory activity was found in the plantlet control and those treated with 4.0 mg/l chitosan of the phenolic and flavonoid fractions. The maximum IC50 (7.36 ± 0.85) was found in the plantlet extract treated with 4.0 mg/l chitosan, followed by the plantlet control, which recorded 11.53 ± 0.86. Using AutoDuck 4.2 and PyMol software, it was revealed that rutin and caffeic acid compounds (the main active components) own hydrogen binding interactions and hydrophobic binding interactions with the active site of alpha-amylase from the porcine pancreas (PPA) enzyme with good binding energy (5.74 and 5.71 kcal/mol, respectively). [ABSTRACT FROM AUTHOR]

Salivary alpha‐amylase (sAA) is an enzyme found in saliva and is considered a noninvasive biomarker for sympathetic nervous system activity. While a wide range of sAA activity in response to stress has been reported in nonhuman primates, the effects of stress on sAA activity in common marmosets are still unknown. We tested the hypothesis that advanced age and cognitive function may have an impact on stress‐related sAA reactivity in marmosets. Thirteen marmosets (nine males and five females) had saliva samples collected during a stressful condition (manual restraint stress) at two different time points, with a 60‐min interval. On the next day, the animals underwent the object recognition test (ORT, a type of cognitive test), and then oral examinations. The animals were categorized into two age groups: old (10–13 years), and very old (15–22 years). Irrespective of age, sAA levels showed a significant difference between T1 (mean 2.07 ± 0.86 U/mL) and T2 samples (mean 1.03 ± 0.67 U/mL), with higher values observed at T1 (p < 0.001). The intra‐assay coefficients of variation (CV) for low and high sAA concentrations were 10.79% and 8.17%, respectively, while the interassay CVs for low and high sAA concentrations were 6.39% and 4.38%, respectively. Oral health issues were common but did not significantly impact sAA levels. The ORT indicated that the animals could recognize an object placed in the cage 6 h after familiarization. In conclusion, all marmosets showed a higher sAA concentration in the first saliva sample as compared to the second saliva sample collected 1 h later, indicating adaptation to stress. No significant differences in sAA levels were observed between sexes, ORT performance, or oral health. Our results indicate that autonomic responsivity and cognitive (memory) functions were preserved even in very old marmosets. Highlights: This study examines how advanced age and cognitive function impact salivary alpha‐amylase (sAA) reactivity.sAA, a stress biomarker, varies in response to stress in common marmosets, largely unexplored till now.Results show higher sAA levels during initial stress, with preserved autonomic and cognitive function in older marmosets. [ABSTRACT FROM AUTHOR]

Mao wine is produced from mao (Antidesma thwaitesianum) and is a local beverage promoting Sakon Nakhon province, Thailand. Understanding the fermentation process is necessary in order to enhance the quality of mao wine products. This study aims to examine the kinetics of mao wine fermentation using 3 commercial types of Saccharomyces cerevisiae yeasts, namely Lalvin 71B, Lalvin RC212 and Lalvin K1- V1116, with an initial pH of 3.4, a sweetness of 22 % Brix and a yeast concentration of 0.4 g L-1 in controlled conditions at 25 °C for 10 days. The results revealed that the kinetic models, including logistic, Luedeking-Piret and modified Luedeking-Piret equations, are appropriate for analyzing the kinetics of yeast growth, alcohol production and reducing sugar, respectively. Lalvin K1-V1116 yeast yielded the highest capacity for alcohol production (~113 g L-1), sugar-to-alcohol conversion (45 % conversion), and product efficiency (88 %), indicating its suitability for mao wine production. Furthermore, the investigation of the alpha-amylase inhibitory activity revealed that the IC50 values of mao wine samples were 2.88 - 4.78 mg mL-1, which were roughly 410 - 680 times greater than those of acarbose. These findings revealed the low efficacy of mao wine in inhibiting alpha-amylase activity. Nevertheless, a decrease in the level of inhibition may result in a decreased likelihood of side effects or undesirable reactions. This treatment is especially beneficial for people who have medication sensitivities. Additional research is required in order to substantiate its applicability in the management of type 2 diabetes. [ABSTRACT FROM AUTHOR]

T2DM is characterized by high blood glucose levels and can result in significant consequences such as nephropathy, neuropathy, retinopathy, and cardiovascular disease. Inhibiting the digestion of dietary carbohydrates is one of the treatment options for managing postprandial hyperglycemia in T2DM. 95% patient have type II diabetes mellitus. Although many synthetic drugs have been identified against pancreatic aamylase. These medications have side effects, and new medications are being developed to address these issues. Managing diabetes without side effects is a significant challenge. This study tested 83 active constituent derived from the plant Abrus precatorius against the human pancreatic aamylase using in-silico computational approaches such as molecular docking and molecular dynamics simulation approaches. Schr€odinger, a drug discovery package with modules applicable for molecular docking, protein-ligand interaction analysis, molecular dynamics studies. Six active constituent, namely, the Vitexin (-10.6kcal/mol), delphinidin-3- 5,diglucoside(-10.5kcal/mol,), abrusin(-9.18kcal/mol), taxifolin-3- glucoside (-8.83 kcal/mol), pelargonidin3-glucoside (-8.72 kcal/mol), and Quercetin (-8.22 kcal/mol ). All the docking score more negative than the control ligand, i.e. Myricetin (–7.3kcal/mol). The molecular dynamics analysis suggested that top two docked compounds, it showed considerable stability within the protein’s active site. As a result, Both compounds found in this work are proposed as promising antidiabetic possibilities and should be evaluated further in vitro and in vivo. [ABSTRACT FROM AUTHOR]

Pistacia atlantica has been widely used as a herbal remedy in the Mediterranean and the Middle East since antiquity. P. atlantica has been used as an antidiabetic drug plant in Arabic folk medicine. This study aims to determine the hypoglycemic and antioxidant activities of the gall part induced by Forda riccobonii in P. atlantica leaves from Algeria. Hypoglycemic activity was determined by measuring alpha-amylase inhibition activity using spectrophotometric analysis of extracts. Antioxidant activity was determined by DPPH, ABTS scavenging assay and FRAP reducing assays. A molecular docking study was carried out on compounds previously isolated from leaves using AutoDock vina software. The highest total phenolic content (655.49±18.36 mg GAE/g DM) and flavonoid content (381.30±15.47 mg QE/g DM) were detected in the butanol fraction. Based on the DPPH assays the ethyl acetate and butanol fractions had high antioxidant activities with EC50 2.46 and 9.94 µg/mL respectively. According to the ABTS assay, the best antioxidant activities were for the 35% MeOH-H2O (4.22 µg/mL) and ethyl acetate fractions (7.23 µg/mL). The a-amylase inhibitory activity was highest in the ethyl acetate fraction (IC50 64.53±2.50 µg/mL) followed by the positive control acarbose (78.98±6.23 µg/mL) and butanol fraction, (151.85±3.01 µg/mL). The in silico study of compounds 5,6,7,4'-tetra-hydroxy-flavonol-3-O-rutinoside and 3',5,7-tri-hydroxy-4'-methoxy-flavanone gave the best docking score (-8,3491 Kcal/mol and -8,1583 Kcal/mol, respectively). Finally, the in silico studies have confirmed significant a-amylase inhibitory activity with these molecules. As well as, the ADMET properties of these molecules appeared to be non-carcinogenic and non-hepatotoxic. Therefore, the gall's extract of P. atlantica could be used in the treatment of diabetes. [ABSTRACT FROM AUTHOR]

BackgroundDysfunction in major stress response systems during the acute aftermath of trauma may contribute to risk for developing posttraumatic stress disorder (PTSD). The current study investigated how PTSD diagnosis and symptom severity, depressive symptoms, and childhood trauma uniquely relate to diurnal neuroendocrine secretion (cortisol and alpha-amylase rhythms) in women who recently experienced interpersonal trauma compared to non-traumatized controls (NTCs).MethodUsing a longitudinal design, we examined diurnal cortisol and alpha-amylase rhythms in 98 young women (n = 57 exposed to recent interpersonal trauma, n = 41 NTCs). Participants provided saliva samples and completed symptom measures at baseline and 1-, 3-, and 6-month follow-up.ResultsMultilevel models (MLMs) revealed lower waking cortisol predicted the development of PTSD in trauma survivors and distinguished at-risk women from NTCs. Women with greater childhood trauma exposure exhibited flatter diurnal cortisol slopes. Among trauma-exposed individuals, lower waking cortisol levels were associated with higher concurrent PTSD symptom severity. Regarding alpha-amylase, MLMs revealed women with greater childhood trauma exposure exhibited higher waking alpha-amylase and slower diurnal alpha-amylase increase.ConclusionsResults suggest lower waking cortisol in the acute aftermath of trauma may be implicated in PTSD onset and maintenance. Findings also suggest childhood trauma may predict a different pattern of dysfunction in stress response systems following subsequent trauma exposure than the stress system dynamics associated with PTSD risk; childhood trauma appears to be associated with flattened diurnal cortisol and alpha-amylase slopes, as well as higher waking alpha-amylase.

General peri- and postnatal characteristics may serve as markers linking pre- or early postnatal events to later health outcomes, which in turn are associated with altered stress- and immune system activity. Our exploratory study investigated whether A) the common perinatal measures “birth weight” and “birth mode” and B) the postnatal characteristics “breastfeeding” and “vaccination status” are associated with markers of stress systems – the hypothalamic-pituitary-adrenal (HPA) axis and autonomous nervous system (ANS) – and inflammation in healthy young adults (n = 68, females: 70.6 %, mean age: 24.21 years, SD = 4.38) exposed to psychosocial challenge, the ‘Trier Social Stress Test’ (TSST). Salivary cortisol, alpha-amylase (sAA) and plasma interleukin-6 (IL-6) were assessed before, during and after the TSST. Participants provided information on peri- and postnatal characteristics. Linear regressions were performed to determine whether peri-/postnatal variables predict basal and stress-response-related biomarker levels. Controlling for sex and sex hormone use as relevant confounders, we found a significant association between birth weight and cortisol recovery (p = 0.032), with higher birth weight predicting higher cortisol recovery values. There were no other significant associations between predictor and outcome variables. Our results show that, in healthy young adults of mixed gender, normal-ranged birth weight is related to the cortisol response to psychosocial stress, indicating a long-term association of this perinatal marker with HPA axis function. In contrast, birth weight was not associated with markers of the ANS stress response or inflammation in adulthood. Our results further suggest that the measures birth mode, duration of breastfeeding, and vaccination status at 4 months of age do not relate to markers of the inflammatory and stress systems in adulthood.

Abstract Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of the alpha‐amylase (Ronozyme® HiStarch CT/L) produced with a genetically modified strain of Bacillus licheniformis (DSM 34315) as a zootechnical feed additive for chickens for fattening, turkeys for fattening and minor growing poultry species. The additive is available in two forms, a coated thermotolerant granulate formulation and a liquid formulation. The production strain and its DNA were not detected in an intermediate concentrated product representative of the final formulations. The Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) concluded that the final product did not trigger safety concerns with regard to the genetic modification. The Panel concluded that Ronozyme® HiStarch CT/L is safe for chickens for fattening, turkeys for fattening and minor growing poultry species at the recommended inclusion level of 80 KNU/kg complete feed. The use of Ronozyme® HiStarch CT/L in animal nutrition under the proposed conditions of use is safe for the consumers and the environment. The additive in any form was shown to be non‐irritant to the skin and the solid form was shown to be non‐irritant to the eyes. No conclusions could be drawn on the potential of the liquid form to be irritant to the eyes or on the potential of both forms of the additive to be dermal sensitisers due to lack of data. Owing to the proteinaceous nature of the active substance, the additive in either form was considered to be a respiratory sensitiser and any exposure by inhalation is considered a risk. In the absence of data, the Panel was not in the position to conclude on the efficacy of Ronozyme® HiStarch CT/L for chickens for fattening, turkeys for fattening and minor growing poultry species.

Eight compounds, including one anthraquinone, two bibenzyls, one phenanthrene, three dihydrophenanthrenes, and one flavonoid, were isolated from the roots of Dendrobium polyanthum Wall. ex Lindl. Among these, six compounds were investigated for inhibitory activities against alpha-glucosidase, alpha-amylase, and advanced glycation end products (AGEs) production. Additionally, molecular docking was conducted to analyze the interactions of the test compounds with alpha-glucosidase. Moscatin, the only isolated phenanthrene, displayed the strongest anti–alpha-glucosidase activity with an IC50 of 32.45 ± 1.04 μM, approximately 10-fold smaller than that of acarbose. Furthermore, moscatilin most strongly inhibited alpha-amylase and AGEs production with IC50 values of 256.94 ± 9.87 and 67.89 ± 9.42 μM, respectively. Molecular docking analysis revealed the effective binding of all substances to alpha-glucosidase with smaller lowest binding energy values than acarbose. Moscatin was selected for kinetics studies, and it was identified as a non-competitive inhibitor with approximately 9-fold greater inhibitory capability than acarbose. This study represents the first report on the phytochemical constituents and antidiabetic potential of compounds derived from the roots of D. polyanthum Wall. ex Lindl.

Using high-throughput metagenomics on commercial microbial fermentation products, DNA from a new unauthorized genetically modified microorganism (GMM), namely the GM B. licheniformis strain producing alpha-amylase (GMM alpha-amylase2), was recently discovered and characterized. On this basis, a new qPCR method targeting an unnatural association of sequences specific to the GMM alpha-amylase2 strain was designed and developed in this study, allowing to strengthen the current GMM detection strategy. The performance of the newly developed qPCR method was assessed for its specificity and sensitivity to comply with the minimum performance requirements established by the European Network of GMO Laboratories for GMO analysis. Moreover, the transferability of the in house validated qPCR method was demonstrated. Finally, its applicability was confirmed by a pilot market surveillance of GMM contaminations conducted for the first time on 40 alpha-amylase food enzyme products labelled as containing alpha-amylase. This pilot market surveillance allowed also to highlight numerous contaminations with GMM alpha-amylase2, including frequent cross-contaminations with other GMM strains previously characterized. In addition, the presence of full-length AMR genes, raising health concerns, was also reported.

Objective(s): This study focused on the evaluation of antioxidant and antidiabetic activities of polyherbal extract (PHE), containing Cassia absus (L.), Gymnema sylvestre (R. Br.), Nigella sativa (L.), and Piper nigrum (L.), in alloxan-induced diabetes model.Materials and Methods: In vitro, HPLC characterization, DPPH scavenging assay, and α-amylase inhibition test were conducted. In vivo, acute oral toxicity of PHE was assessed. Alloxan-induced diabetic Wistar rats (n=6) were orally treated with PHE (200, 400, and 600 mg/kg/day) and glibenclamide (GLB; 10 mg/kg/day) for six consecutive weeks. Then, biochemical biomarkers, oxidative stress parameters, histopathological examination, and mRNA expression levels (RT-qPCR) were determined.Results: The presence of polyphenols in PHE was confirmed in correlation to marked DPPH scavenging (IC50: 1.60 mg/ml) and α-amylase inhibition (IC50: 0.82 mg/ml). PHE demonstrated no toxicity in rats up to a dose of 2000 mg/kg. In diabetic rats, PHE dose-dependently ameliorated the serum levels of glucose, insulin, glycated hemoglobin A1c (HbA1c), leptin, and glucokinase (GCK). Also, PHE substantially alleviated serum inflammatory markers (TNF-α and CRP) and oxidative stress indicators (MDA, SOD, and CAT) in pancreatic tissues. PHE, particularly at 600 mg/kg, attenuated cellular oxidative stress via modulating the mRNA expression levels of genes regulating MAPK/JNK (Mapk-8, Traf-4, and Traf-6) and Nrf-2/Keap-1 pathways and promoted insulin signaling through up-regulating insulin signaling cascade (Pdx-1, Ins-1, and Ins-2), as compared to GLB. Furthermore, histopathological findings supported the aforementioned results.Conclusion: Our study suggests that polyherbal extract has promising antioxidant and antidiabetic activities by modulating the MAPK/JNK, Nrf-2/Keap-1, and insulin signaling pathways.

Subject of study Coffee infusions are extremely popular worldwide due to their pleasant taste, ability to reduce fatigue, increase concentration and improve productivity. Infusion extracted from coffee beans are mixtures of many chemical compounds, both volatile and non-volatile. Among the non-volatile components, caffeine, trigonelline and chlorogenic acid appear to be the dominant ones. These substances are responsible for both the flavour sensations and influence on biological activities, such as mental stimulation effects or reducing the risk of type 2 diabetes. Aim of study The aim of the present study was: (I) to develop and validate a high-performance thin-layer chromatography (HPTLC) method for the simultaneous quantification of caffeine, trigonelline and chlorogenic acid in coffee bean infusions; (II) to use this method to determine caffeine, trigonelline and chlorogenic acid in green, light and dark roasted coffee bean infusions prepared both by immersion (brewing) and pressure methods; (III) to evaluate the effects of coffee infusions on α-amylase activity. Materials and methods Simultaneous determination of caffeine, trigonelline and chlorogenic acid was carried out on glass HPTLC plates coated with silica gel 60 F254 with a concentration zone and the mobile phase consisted of a mixture of: chloroform, ethyl acetate, methanol and formic acid in a volume ratio of 10:6:3:1 (v/v). After the plates were developed and dried, densitometric scanning was performed at wavelengths of 270, 275 and 330 nm for trigonelline, caffeine and chlorogenic acid, respectively. The analysis of the effect of coffee infusions on the α-amylase activity was carried out using a porcine pancreas α-amylase. Results Green coffee infusions prepared using the immersion method was proved to have the highest chlorogenic acid content, while the highest trigonelline and caffeine contents were observed in infusions also obtained using the immersion method, but from dark-roasted coffee. Infusions of green coffee and light and dark roasted coffee inhibited α-amylase activity, with green coffee infusions showed the strongest effect. Conclusions The developed method is selective, precise, accurate and sensitive, so it can be used for the simultaneous determination of caffeine, trigonelline and chlorogenic acid in heat-treated and non-heat-treated coffee bean infusions. The results of the analysis of the effect of coffee infusions on the α-amylase activity create the opportunity to use these infusions, particulary unroasted coffee infusions, in the prevention of type 2 diabetes.

Background and purpose: Diabetes mellitus is a persistent hyperglycemic condition. Thai cuisine and medicine incorporate spices: nutmeg, mace, clove buds, cardamom, cinnamon, and coriander. The in vitro impacts of these spices on anti-diabetic, antioxidant, anti-inflammatory, and total phenolic and flavonoid content were assessed. Experimental approach: Alpha-amylase and alpha-glucosidase inhibition assays were conducted. Antioxidant potential was measured through DPPH and ABTS assays. Anti-inflammatory activity was determined by inhibiting nitric oxide generation in RAW 264.7 cells. Total phenolic content was quantified using the Folin Ciocalteu method, while total flavonoid content was estimated via the aluminum chloride colorimetric method. Findings/Results: Ethanolic and aqueous extracts of a blend of spices (Siam cardamom, nutmeg, mace, and clove buds), denoted as 4-GlurE and 4-GlurA, displayed concentration-dependent inhibition of alpha- glucosidase, with IC50 values of 0.373 and 0.435 mg/mL, respectively. 4-GlurE and 4-GlurA exhibited antioxidant activity, by ABTS·+ radical and DPPH scavenging capabilities. 4-GlurE demonstrated anti-inflammatory potential by reducing nitric oxide generation (IC50: 43.95 ± 2.47 μg/mL). 4-GlurE and 4-GlurA possessed total phenolic content (TPC) of 122.47 ± 1.12 and 148.72 ± 0.14 mg GAE/g, respectively. 4-GlurE exhibited a higher total flavonoid content (TFC) compared to the aqueous extract (340.33 ± 4.77 and 94.17 ± 3.36 mg QE/g). Cinnamon and clove aqueous extracts were more potent than acarbose in alpha-glucosidase inhibition with the highest antioxidant activity. Polyphenol levels (TPC and TFC) exhibited strong correlations with antioxidant capacity. Conclusions and implications: Findings are consistent with the traditional use of 4-Glur, with cinnamon, for diabetes prevention and treatment.

Abstract Background Type 2 Diabetes mellitus (DM) is an affliction impacting the quality of life of millions of people worldwide. An approach used in the management of Type 2 DM involves the use of the carbohydrate-hydrolyzing enzyme inhibitor, acarbose. Although acarbose has long been the go-to drug in this key approach, it has become apparent that its side effects negatively impact patient adherence and subsequently, therapeutic outcomes. Similar to acarbose in its mechanism of action, bee propolis, a unique natural adhesive biomass consisting of biologically active metabolites, has been found to have antidiabetic potential through its inhibition of α-amylase. To minimize the need for ultimately novel agents while simultaneously aiming to decrease the side effects of acarbose and enhance its efficacy, combination drug therapy has become a promising pharmacotherapeutic strategy and a focal point of this study. Methods Computer-aided molecular docking and molecular dynamics (MD) simulations accompanied by in vitro testing were used to mine novel, pharmacologically active chemical entities from Egyptian propolis to combat Type 2 DM. Glide docking was utilized for a structure-based virtual screening of the largest in-house library of Egyptian propolis metabolites gathered from literature, in addition to GC–MS analysis of the propolis sample under investigation. Thereafter, combination analysis by means of fixed-ratio combinations of acarbose with propolis and the top chosen propolis-derived phytoligand was implemented. Results Aucubin, identified for the first time in propolis worldwide and kaempferol were the most promising virtual hits. Subsequent in vitro α-amylase inhibitory assay demonstrated the ability of these hits to significantly inhibit the enzyme in a dose-dependent manner with an IC50 of 2.37 ± 0.02 mM and 4.84 ± 0.14 mM, respectively. The binary combination of acarbose with each of propolis and kaempferol displayed maximal synergy at lower effect levels. Molecular docking and MD simulations revealed a cooperative binding mode between kaempferol and acarbose within the active site. Conclusion The suggested strategy seems imperative to ensure a steady supply of new therapeutic entities sourced from Egyptian propolis to regress the development of DM. Further pharmacological in vivo investigations are required to confirm the potent antidiabetic potential of the studied combination.