Α-amylase Inhibitory Activity Research Articles

In Vitro Hypoglycemic and Antioxidant Activities of Dichloromethane Extract of Xerophyta spekei.

Diabetes mellitus is a chronic metabolic disorder which has greatly led to an increase in morbidity and mortality globally. Although Xerophyta spekei is widely used for the management of diabetes among the Embu and Mbeere communities in Kenya, it has never been empirically evaluated for its hypoglycemic activity. This study was carried out to verify the hypoglycemic activity of dichloromethane (DCM) extract of Xerophyta spekei as well as its antioxidant activity using various in vitro techniques. Phytochemicals associated with its antioxidant activity were identified through GC-MS. Data were subjected to descriptive statistics and expressed as mean ± standard error of the mean (X̄ ± SEM). Comparison between various variables was performed by using unpaired Student's t-test and one-way analysis of variance (ANOVA), followed by Tukey's post-hoc test. The confidence interval was set at 95%. The obtained results were presented in tables and graphs. Results showed that there was no difference in α-amylase inhibition activity between the plant extract and the standard (IC50 525.9 ± 12.34 and 475.1 ± 9.115, respectively; p > 0.05). Besides, the glucose adsorption activity of the extract increased with an increase in glucose concentration (from 5.89 to 32.64 mg/dl at 5 mmol and 30 mmol of glucose, respectively; p < 0.05). The extract also limited the diffusion of glucose more than the negative control (7.49 and 17.63 mg/dl, respectively; p < 0.05). It also enhanced glucose uptake by yeast cells. In addition, the studied plant extract showed notable antioxidant activities. The therapeutic effects exhibited by this plant in managing diabetes mellitus and other ailments could be due to its antioxidant as well as its hypoglycemic activity. The study recommends the evaluation of X. spekei for in vivo hypoglycemic and antioxidant activities. Besides, the isolation of bioactive phytochemicals from the plant may lead to the development of new hypoglycaemic agents.

Mixed fermentation of navel orange peel by Trichoderma viride and Aspergillus niger: Effects on the structural and functional properties of soluble dietary fiber

Navel orange peel is a potential source of soluble dietary fiber. In order to make full use of this resource and improve its physiological activities, the effects of single and mixed strains fermentation modifications were compared in this study. The structural, physicochemical, and functional properties of soluble dietary fiber fermented by Trichoderma viride and/or Aspergillus niger were investigated. The results showed that mixed fermentation resulted in significantly higher activity of complex cellulase than single fermentation. On the second day, when T. viride and A. niger were inoculated at a ratio of 3:1, the activities of carboxymethyl cellulase, β-glucosidase, and filter paper lyase were 1.54, 1.61, and 1.30 times those of single fermentation. Mixed fermentation increased the yield of soluble dietary fiber from 6.63% to 15.73%. Scanning electron microscopy and Fourier transform infrared spectroscopic analyses suggested that soluble dietary fiber produced by mixed fermentation had smaller particle size, lower crystallinity, and more disrupted structure. Moreover, the functional properties of soluble dietary fiber were remarkably improved. After mixed fermentation, the glucose and cholesterol adsorption amounts, and α-amylase inhibition activity increased by 1.51, 1.63, and 1.76 times, respectively. In summary, mixed fermentation was an efficient and environmentally friendly modification method. Modified SDF can be utilized in the food processing industry, expanding the range of by-product applications.

α-glucosidase and α-amylase inhibitory activity of flavonols from Stenochlaena palustris (Burm.f.) Bedd

Stenochlaena palustris (Burm.f.) Bedd., a fern species native to India, Southeast Asia, Polynesia, and Australia, has a long history of medical including as a diabetic therapy. This study aimed to isolate bioactive compounds from S. palustris ethyl acetate extract and assess their in vitro and in silico inhibitory activities against α-glucosidase and α-amylase. The successful separation of five flavonols, namely stenopalustroside A (1), tiliroside (2), kaempferol (3), quercetin (4), and rutin (5), was achieved through phytochemical analysis. The compounds exhibited a range of inhibitory activities against α-glucosidase and α-amylase, with IC50 values ranging from 40 to 250 µg/mL. Notably, the biological activities of compound 1 have been reported for the first time. Compound 4 was the most effective inhibitor of both enzymes among the isolated compounds. Studies performed in silico reveal that the interactions between amino acids in compounds 4 and 5 are remarkably comparable to those observed in the positive control. These compounds share this commonality, and as a result, they both have the potential to be active agents.

Phytochemical Analysis and Biological Activities of Ficus racemosa Linn., Myrica esculenta Buch-Ham. Ex D. Don and Urtica dioica Linn.

Medicinal plants like Ficus racemosa, Myrica esculenta, and Urtica dioica were traditionally used to treat diabetes, cough, fever etc. These plants were collected from Kathmandu and Gorkha district of Nepal and subjected to various biological activities. Methanol extract of all three selected plants when subjected to phytochemical analysis showed the presence of various chemical constituents such polyphenols, flavonoids, tannins, saponins, and glycosides. The methanol extract of F. racemosa bark was found with strongest DPPH radical neutralization activity having IC50 value of 41.48 µg/mL, which was comparable with standard ascorbic acid (31.07 µg/mL). The bark of F. racemosa contained the highest level of total phenolic and total flavonoid contents with the values of 168.89 ± 1.08 mg GAE/g and 41.56 ± 1.83 mg QE/g respectively. The methanol extract of bark of F. racemosa showed high α-amylase inhibitory activity with IC50 value of 164.99 μg/mL. Brine shrimp bioassay showed that the methanol extracts of bark of F. racemosa, bark of M. esculenta, leaves of M. esculenta and U. dioica were lethal to brine shrimp nauplii with LC50 values of 70.79, 177.83, 190.54 and 269.15 µg/mL respectively. Methanol extract of leaves of U. dioica was least active in these biological activities.

Antioxidant activity of Phellinus igniarius fermentation mycelia contributions of different solvent extractions and their inhibitory effect on α-amylase

Phellinus spp. have historically been used as traditional medicines to treat various diseases owing to their antioxidant, antitumor, and antidiabetic activities. Polysaccharides exhibit antidiabetic activity. In the present study, the polysaccharide contents of four Phellinus strains were compared. Phellinus igniarius QB72 possessed higher polysaccharide production, stronger 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, and α-amylase inhibitory activity. The three polysaccharides were sequentially extracted and partially purified from the fermentation mycelia using hot water, 1 % (NH4)2C2O4, and 1.25 M NaOH. Hot water extract polysaccharides exhibited higher DPPH radical scavenging and strong inhibitory activity against α-amylase with an IC50 value of 6.84 ± 0.37 mg/mL. The carbohydrate content of A1 (approximately 17457 Da) was approximately 88.28 %. The α-amylase inhibitory activity IC50 was decreased (3.178 ± 0.187 mg/mL) after DEAE water elution. P. igniarius QB72 hot-water extracts of partially purified polysaccharides have great potential as α-amylase inhibitors in food and medication-assisted additives.

Chemical Profiling, in-vitro biological evaluation and molecular docking studies of Ruellia tweediana: An unexplored plant

Many Ruellia species have been utilized in traditional medicine and despite the prevalent use of Ruellia tweediana in folk medicine, its antioxidant potential and polyphenol content have not been investigated. Therefore, the present study aimed to explore the medicinal value of R. tweediana by evaluating its total phenolic (TPC) and flavonoid contents (TFC), GC–MS analysis, antioxidant, antibacterial, and enzyme inhibition activities. The TPC and TFC of the extract/fractions were assessed using the Folin-Ciocalteu and aluminum trichloride methods, respectively. To determine the antioxidant capacity, five different assays were used: DPPH, ABTS, CUPRAC, FRAP, and metal chelating assays. The inhibition activity against α-glucosidase, α-amylase, cholinesterases, and lipoxygenase enzymes was also analyzed. Furthermore, GC–MS was performed for chemical screening of non-polar fraction. The methanol extract showed the maximum TPC (167.34 ± 2.23 mg GAE/g) and TFC (120.43 ± 1.71 mg RE/g) values among all the tested samples. GC–MS screening of the n-hexane fraction showed the presence of 40 different phytoconstituents. The results demonstrated the highest scavenging potential of the methanol extract against DPPH (167.79 ± 2.75 mg TE/g) and ABTS (255.32 ± 2.91 mg TE/g) radicals, as well as the metal-reducing capacity measured by CUPRAC (321.34 ± 3.09 mg TE/g), FRAP (311.32 ± 2.91 mg TE/g), and metal chelating assay (246.78 ± 10.34 mg EDTAE/g). Notably, the n-hexane fraction revealed the highest α-glucosidase and α-amylase inhibition activity (186.8 ± 2.84 and 179.7 ± 4.32 mg ACAE/g, respectively) while methanol extract showed highest acetylcholinesterase and butyrylcholinesterase inhibition activity (198.6 ± 3.31 and 184.3 ± 2.92 mg GALE/g, respectively). The GC–MS identified Lupeol showed best binding affinity with all docked enzymes as compared to standard compounds. The presence of bioactive phytoconstituents showed by GC–MS underscores the medicinal importance of R. tweediana, making it a promising candidate for natural medicine.

Chemical Analysis of the Antihyperglycemic, and Pancreatic α-Amylase, Lipase, and Intestinal α-Glucosidase Inhibitory Activities of Cannabis sativa L. Seed Extracts.

Cannabis is considered (Cannabis sativa L.) a sacred herb in many countries and is vastly employed in traditional medicine to remedy numerous diseases, such as diabetes. This research investigates the chemical composition of the aqueous extracts from Cannabis sativa L. seeds. Furthermore, the impact of these extracts on pancreatic α-amylase and lipase, and intestinal α-glucosidase enzymes is evaluated, as well as their antihyperglycemic effect. Analysis of the chemical composition of the aqueous extract was conducted using high-performance liquid chromatography with a photodiode array detector (HPLC-DAD). In contrast, the ethanol, hexanic, dichloromethane, and aqueous extract compositions have been established. Additionally, the inhibitory effects of ethanolic, dichloromethane, and aqueous extracts on pancreatic α-amylase and lipase, and intestinal α-glucosidase activities were evaluated in vitro and in vivo. The results of HPLC analysis indicate that the most abundant phenolic compound in the aqueous cannabis seed extract is 3-hydroxycinnamic acid, followed by 4-hydroxybenzoic acid and rutin acid. Moreover, administration of ethanolic and aqueous extracts at a dose of 150 mg/Kg significantly suppressed postprandial hyperglycemia compared to the control group; the ethanolic, dichloromethane, and aqueous extracts significantly inhibit pancreatic α-amylase and lipase, and intestinal α-glucosidase in vitro. The pancreatic α-amylase test exhibited an inhibition with IC50 values of 16.36 ± 1.24 µg/mL, 19.33 ± 1.40 µg/mL, 23.53 ± 1.70 µg/mL, and 17.06 ± 9.91 µg/mL for EAq, EDm, EET, and EHx, respectively. EET has the highest inhibitory capacity for intestinal α-glucosidase activity, with an IC50 of 32.23 ± 3.26 µg/mL. The extracts inhibit porcine pancreatic lipase activity, demonstrating their potential as lipase inhibitors. Specifically, at a concentration of 1 mg/mL, the highest inhibition rate (77%) was observed for EDm. To confirm these results, the inhibitory effect of these extracts on enzymes was tested in vivo. The oral intake of aqueous extract markedly reduced starch- and sucrose-induced hyperglycemia in healthy rats. Administration of the ethanolic extract at a specific dose of 150 mg/kg significantly reduced postprandial glycemia compared with the control group. It is, therefore, undeniable that cannabis extracts represent a promising option as a potentially effective treatment for type 2 diabetes.

Fluorinated thiazole–thiosemicarbazones hybrids as potential PPAR-γ agonist and α-amylase, α-glucosidase antagonists: Design, synthesis, in silico ADMET and docking studies and hypoglycemic evaluation

In addition to Peroxisome Proliferator-Activated Receptor-gamma (PPAR-γ), which is essential for controlling type-2 diabetes mellitus, α-amylase and α-glucosidase inhibition is a key strategy in managing postprandial hyperglycemia. In this work a series of 2-(1-(2-((4-fluorophenyl)amino)-4-methylthiazol-5-yl)ethylidene)hydrazine-1-carbothioamide derivatives (3–6) was designed and synthesized through theiosemicarbazide derivatives, the structural elucidation of the new synthesized compounds was established using elemental and spectroscopic analyses. The antidiabetic activities of the new thiazole-thiosemicarbazone hybrids were screened in vivo for blood glucose lowering activity in normal rats and compared with pioglitazone as antidiabetic standard. Male Sprague-Dawley rats had reduced blood sugar levels after exposure to compounds 5 and 6 compared to pioglitazone. Additionally, the PPAR-γ agonist activity of compounds 5 and 6 as well as their ability to inhibit α-amylase and α-glucosidase were assessed. When compared to standard acarbose, which had an IC50 value of 0.74 ± 0.15 mM/mL, compound 5 showed the highest α-glucosidase inhibitory activity with IC50 value of 0.446 ± 0.01 mM/mL, while compound 6 showed good activity with an IC50 value of 1. 914 ± 0.04 mM/mL. However, compound 5 and compound 6 had weak α-amylase inhibitory activity, with IC50 values of 11.78 ± 0.58 and 24.18 ± 1.2 µg/mL respectively. Furthermore, Peroxisome proliferator activated receptor γ [PPAR-γ] agonist activity for compound 5 and compound 6 was assessed and gave promising result as [PPAR-γ] agonist with IC50 value of 0.938 ± 0.023 and 0.947 ± 0.024 ng/mL respectively superior to that of pioglitazone with IC50 value of 1.912 ± 0.11 ng/mL. All the results supported compound 5 as promising candidates for construction of new antidiabetic agents. Also, in silico ADMET and docking studies for the most promising derivatives 5, 6, Acarbose, and Pioglitazone were done.

Antifungal, Antimycobacterial, Protease and α‒Amylase Inhibitory Activities of a Novel Serine Bifunctional Protease Inhibitor from Adenanthera pavonina L. Seeds.

Antifungal resistance poses a significant challenge to disease management, necessitating the development of novel drugs. Antimicrobial peptides offer potential solutions. This study focused on extraction and characterization of peptides from Adenanthera pavonina seeds with activity against Candida species, Mycobacterium tuberculosis, proteases, and α-amylases. Peptides were extracted in phosphate buffer and heated at 90°C for 10 min to create a peptide rich heated fraction (PRHF). After confirming antimicrobial activity and the presence of peptides, the PRHF underwent ion exchange chromatography, yielding retained and non-retained fractions. These fractions were evaluated for antimicrobial activity and cytotoxicity against murine macrophages. The least toxic and most active fraction underwent reversed-phase chromatography, resulting in ten fractions. These fractions were tested for peptides and antimicrobial activity. The most active fraction was rechromatographed on a reversed-phase column, resulting in two fractions that were assessed for antimicrobial activity. The most active fraction revealed a single band of approximately 6 kDa and was tested for inhibitory effects on proteases and α-amylases. Thermal stability experiments were conducted on the 6 kDa peptide at different temperatures followed by reassessment of antifungal activity and circular dichroism. The 6 kDa peptide inhibited yeasts, M. tuberculosis, human salivary and Tenebrio molitor larvae intestine α-amylases, and proteolytic activity from fungal extracts, and thus named ApPI. Remarkably, ApPI retained antifungal activity and conformation after heating and is primarily composed of α-helices. ApPI is a thermally stable serine protease/α-amylase inhibitor from A. pavonina seeds, offering promise as a foundational molecule for innovative therapeutic agents against fungal infections and tuberculosis.

Exploring the potential of Lindernia ciliata: isolation, characterization, and pharmacological evaluation of its anti-diabetic properties

We isolated two phytoconstituents using bioassay guided isolation for anti-diabetic property from the hydroalcoholic extract of Lindernia ciliata (Colsm.) Pennell (Family: Linderniaceae). We assessed the anti-diabetic potential using various assays, including the glucose absorption assay, glucose uptake in isolated rat abdominal muscle assay, insulin secretion by RIN-5F cells assay, α-amylase inhibition activity, and DPP-4 inhibition assay. The results from our study indicated that the ethanol and water fractions obtained from the hydroalcoholic extract significantly improved glucose uptake, demonstrating promising anti-diabetic properties. Further investigation led to the isolation of two distinct compounds, LCF-1 (IUPAC name: 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-4H-chromen-4-one) (LCF-1) and LCF-2 (IUPAC name: 3S,10R,13R,17R)-17-[(2R,5R)-5-ethyl-6-methylheptan-2-yl]-10,13-dimethyl-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-ol). Consequently, our findings suggest that LCF-1 and LCF-2 are two phytoconstituent derived from the hydroalcoholic extract of L. ciliata, could serve as a potential source of anti-diabetic agents. This highlights its suitability for further development in the creation of herbal pharmaceuticals for diabetes treatment.

Decoding the alpha-amylase inhibitory activity of Garcinia indica Choisy by computational and experimental studies

Garcinia indica Choisy is traditionally used to manage diabetes mellitus, also used in various herbal formulations, and is indicated as an insulin sensitizer. The present study investigates the hydroalcoholic extract of G. indica Choisy rind as an α-amylase inhibitor, followed by the identification of the probable lead hits via a computational approach. LC-MS analyse were carried out. In vitro assays for alpha amylase were performed at concentrations of G. indica ranging from (1000–125 µg/mL) and acarbose (25–1.56 µg/mL) with inhibition times ranging from 5th to 30th min. In silico study was performed based upon the bioactives traced from LC-MS data retrieved from the PubChem database and predicted for probable targets of diabetes mellitus, followed by a drug-likeness score, probable side effects, and an ADMET profile. In addition, molecular docking and the dynamics of the representative complex were explored using AutoDock Vina and Gromac. In vitro study revealed IC50 values of 400.5, 526.07, 728.05, 792.7, 848.3, and 897.5 µg/mL and acarbose displayed IC50 values of <1.56, 6.6, 10.46, 12.6, 14.14, and 15.24 µg/mL respectively. LC-MS analysis identified 17 bioactives, of which 13 showed positive druglikness; among them, 5 bioactives were predicted to modulate AMY2A. The gene set enrichment analysis revealed that AMY2A and gambogic acid possessed high edge counts, with the lowest binding energy of gambogic acid- AMY2A (-11.7 kcal/mol), followed by amentoflavone (-10.6 kcal/mol), volkensiflavone (-9.3 kcal/mol), fukugcide (-8.8 kcal/mol), and garcinol (-7.7 kcal/mol) when compared with acarbose- AMY2A (-7.7 kcal/mol), which also indicates high stability up to 100 ns in molecular simulation. Hydroalcoholic extract of G. indica rind α-amylase inhibitory activity, may be due to the presence of gambogic acid and other bioactives. Thus, it can be speculated that G. indica may possess anti-diabetic activity by reducing the post prandial glucose level. This needs to be further confirmed via enriched fractions or pure substances with inhibitory activity to develop Traditional medicine based new drugs

In‐Vitro Investigation of the α‐Amylase Inhibition Activity of Bare Bis‐Benzylidene‐Cyclohexanone Synthesized by a Highly Selective Solvent‐Free Route

AbstractCurrent work reports the highly selective, solvent‐free synthesis of an endorsed bioactive compound, Bis benzylidine cyclohexanone (BBC) through solid acid catalysed cross aldol condensation route and checks its in‐vitro bio activity. The catalytic support (Multiwalled carbon nanotube) employed was synthesized through the highly sophisticated catalytic chemical vapor deposition (CVD) method and simple mechanical grinding strategy was adopted to decorate sulfonic acid on the support. The C1s X‐ray photoelectron peak of at 290.3 eV confirms the effective interaction of sulfonic acid with MWCNT. The sharp and intense desorption peaks observed at approximately 528.7 °C and 655.15 °C in the TPD analysis unmistakably substantiate the strong acidity of the synthesized system. The alpha amylase inhibition activity of the synthesized compound was calculated to be around 88.5 %, which is in par with the commercial drug as it could inhibit only 96 %. Further, the in‐silico (Docking and Molecular Dynamic Simulation) investigations unveiled a new target site for the compound and this can further be studied in detail to advance the applications in drug design. Detailed scrutiny of various parameters was conducted to validate the bioactivity and pharmaceutical potential of the synthesized compound.

Comparative metabolic study of the chloroform fraction of three Cystoseira species based on UPLC/ESI/MS analysis and biological activities

This study aims to investigate the phytoconstituents of the chloroform fraction of three Cystoseira spp. namely C. myrica, C. trinodis, and C. tamariscifolia using UPLC/ESI/MS technique. The results revealed the identification of 19, 20 and 11 metabolites in C. myrica, C. trinodis, and C. tamariscifolia, respectively mainly terpenoids, flavonoids, phenolic acids and fatty acids. Also, an in vitro antioxidant study using FRAP and DPPH assays was conducted where the chloroform fraction of C. trinodis displayed the highest antioxidant activity in both assays, which would be attributed to its highest total phenolics and total flavonoids. Besides, the investigation of COX-1, α-glucosidase and α-amylase inhibitory activities were performed. Regarding C. trinodis, it showed the strongest inhibitory activity towards COX-1. Moreover, it showed potent inhibitory activity towards α-glucosidase and α-amylase enzymes. According to the molecular docking studies, the major compounds characterised showed efficient binding to the active sites of the target enzymes.

Argan: Phytochemical profiling and evaluation of the antioxidant, hypoglycemic, and antibacterial properties of its fruit pulp extracts

Herein, we isolated three triterpenoid saponins from the methanol extract of the fruit pulp of argan. The structures of the identified compounds were determined using comprehensive NMR spectroscopy analyses (1H, 13C NMR, COSY, TOCSY, ROESY, and HSQC), combined with mass spectroscopy. Gas chromatography (GC) was utilized to determine the monosaccharide contents after the samples underwent methanolysis and their glycoside configuration was proved via their trimethylsilyl derivatives. Furthermore, the methanol extract of the fruit pulp and its n-butanol fraction were evaluated for their antioxidant properties via DPPH, ABTS, and FRAP assays, antidiabetic activity using α-amylase and α-glucosidase inhibition activities, and antibacterial properties utilizing microdilution and antibiofilm assays. Compared to the crude methanol extract, our results showed that the n-butanol fraction exhibited more potent antioxidant activity and antibacterial potential against Staphylococcus aureus, Escherichia coli, Salmonella typhi, Enterococcus faecalis, and Pseudomonas aeruginosa (MIC = 12.5–50 mg/mL); while no effect on the bacterial biofilm was observed. The methanol extract was more effective in inhibiting α-glucosidase (EC50 = 0.15 mg/mL), however, the n-butanol fraction elicited strong α-amylase inhibition (EC50 = 0.49 mg/mL). These findings suggest that the fruit pulp of argan could serve as a potential source of phytochemicals suitable for the treatment of diabetes and its related complications.

Evaluation of new candidate probiotic lactobacillus strains isolated from a traditional fermented food- multigrain-millet dosa batter

The food industry is constantly developing new products to meet the increasing global demand of fermented foods, for its health benefits. In this study, the potential probiotics in traditional fermented food - “multigrain-millet” based Dosa batter, and their significant probiotic characteristics was investigated. In addition, the ability of probiotic Lactobacillus to prevent foodborne pathogens was explored. A total collection of 70 isolates were examined from the batter. The six isolates with the best in vitro probiotic characteristics were studied. The six isolates MYSDS57, MYSDS68, MYSDV3, MYSDV2, MYSDV5 and MYSDV6 were identified as Lactiplantibacillus plantarum, Lacticaseibacillus paracasei, Lactobacillus pentosus, Lactiplantibacillus plantarum, Lactiplantibacillus plantarum and Limosilactobacillus fermentum respectively based on 16 S rRNA gene sequencing analysis. All six Lactobacillus were able to tolerate acidic pH (45 %–80 % survivability) and 0.3% bile salt (50 %–80.4 % survivability) up to 5 h of incubation. All tested strains showed higher growth at 37 °C, resistance to different concentrations of phenols (53.3 %–83.6 % of viability) and NaCl (38.2%–49.5% of viability). However, the strains exhibited resistance to antibiotic vancomycin, but no hemolytic and DNase activity. Moreover, six strains showed significantly high values of autoaggregation (54.6%-78.8 %) and cell surface hydrophobicity (70 %-81.7%). The α-glucosidase and α-amylase inhibitory activity ranged from 15.08% to 67.22 % and 50.15%–92.13% respectively. Additionally, the intracellular cell-free supernatant of all six Lactobacillus demonstrated antibacterial activity with the inhibition rate of 64.8 %–86 % (Salmonella typhi), 51.8 %–95.2 % (Escherichia coli), 59.9 %–76.6 % (Listeria monocytogenes), 39.6% −74.3 % (Salmonella paratyphi), 57.4 %–81.3 % (Klebsiella pneumonia), 44 %-67.1% (Staphylococcus aureus) and 7.2 %–54.7 % (Pseudomonas aeruginosa). The strains exhibited higher viability (5.8–7.5 log CFU/mL) to simulated gastric juice, high adhesion to chicken crop epithelial cells and buccal cells. These isolates also reduced the cholesterol level in vitro. Overall, the selected Lactobacillus can be considered promising probiotic candidates for future applications.

Gymnemagenin a Promising Drug Candidate for Management of Hyperglycemia: In-Vitro and In-Vivo Study

Background: Diabetes mellitus is a chronic metabolic disorder resulting from absolute or relative impairments in β-cell functions, insulin resistance, or both and has been exceptionally increasing worldwide. Gymnema sylvestre is an important traditional plant used for the treatment and management of diabetes. Gymnemagenin is an aglycone part of gymnemic acids which is a bioactive component of G. sylvestre. The objective of the present study was to undertake in-vitro and in-vivo studies to generate a stronger biochemical rationale for the management of diabetes mellitus. Results: The result of the preliminary in-vitro study of gymnemagenin suggested further evaluation of the inhibition of key enzymes related to carbohydrate metabolism. Gymnemagenin showed strong α-amylase (IC50 1.17±0.02 mg/mL) and strong α-glucosidase (IC50 2.04±0.17mg/mL) inhibitory activity. The positive in-vitro enzyme inhibition tests paved the way for confirmatory in-vivo studies. The in-vivo studies demonstrated that gymnemagenin (20 mg/kg, b.w.) given orally significantly (P&lt;0.01) reduced area under curve (AUC) in mice when challenged with oral administration of starch and sucrose separately. The reduction in AUC by gymnemagenin was comparable to that of acarbose (10 mg/kg, b.w., p.o.). Conclusion: The research findings clearly supported the traditional use of G. sylvestre in the treatment of diabetes mellitus.

Transcription factor OsMYB30 increases trehalose content to inhibit α-amylase and seed germination at low temperature.

Low-temperature germination (LTG) is an important agronomic trait for direct-seeding cultivation of rice (Oryza sativa). Both OsMYB30 and OsTPP1 regulate the cold stress response in rice, but the function of OsMYB30 and OsTPP1 in regulating LTG and the underlying molecular mechanism remains unknown. Employing transcriptomics and functional studies revealed a sugar signaling pathway that regulates seed germination in response to low temperature (LT). Expression of OsMYB30 and OsTPP1 was induced by LT during seed germination, and overexpressing either OsMYB30 or OsTPP1 delayed seed germination and increased sensitivity to LT during seed germination. Transcriptomics and qPCR revealed that expression of OsTPP1 was upregulated in OsMYB30-overexpressing lines but downregulated in OsMYB30-knockout lines. In vitro and in vivo experiments revealed that OsMYB30 bound to the promoter of OsTPP1 and regulated the abundance of OsTPP1 transcripts. Overaccumulation of trehalose (Tre) was found in both OsMYB30- and OsTPP1-overexpressing lines, resulting in inhibition of α-amylase 1a (OsAMY1a) gene during seed germination. Both LT and exogenous Tre treatments suppressed the expression of OsAMY1a, and the osamy1a mutant was not sensitive to exogenous Tre during seed germination. Overall, we concluded that OsMYB30 expression was induced by LT to activate the expression of OsTPP1 and increase Tre content, which thus inhibited α-amylase activity and seed germination. This study identified a phytohormone-independent pathway that integrates environmental cues with internal factors to control seed germination.

Machine learning combined with molecular simulations to screen α-amylase inhibitors as compounds that regulate blood sugar

Diabetes, a metabolic disease characterized by hyperglycemia, seriously endangers the health and the lives of people. α-Amylase inhibitors have become effective substances to control blood glucose, and attracted extensive attention. In this study, a database of α-amylase inhibitors derived from naturally active small molecules in food was created and a quantitative structure-activity relationship model was developed by combining three machine learning methods (SVM, RF, and LDA) with four descriptors (MOE, ChemoPy, Mordred, and Rdkit). Hydrogen bond and hydrophobic interaction in the inhibition of α-amylase activity was confirmed by molecular docking. Enzyme inhibition experiments showed that the predicted compound had α-amylase inhibitory activity. Nevadensin was identified as a promising candidate of α-amylase inhibitors. The stability of α-amylase binding reaction was verified by molecular dynamics simulation. Optimal process conditions for the extraction of nevadensin from L. pauciflorus maxim were derived from single-factor experiments and response surface modeling. A promising method for digging natural α-amylase inhibitors was developed and the mode between inhibitors and α-amylase was explained in this research.

Effect of Yogurt Ice Cream on the Viability and Antidiabetic Potential of the Probiotics Lactobacillus acidophilus, Lacticaseibacillus rhamnosus, and Bifidobacterium animalis subsp. lactis after In Vitro Digestion.

Probiotics can ameliorate type 2 diabetes mellitus (T2DM) via several mechanisms such as by decreasing inflammatory cytokines and increasing pancreatic β-cell functions. Another targeted mechanism for managing T2DM involves inhibiting α-amylase and α-glucosidase, which exhibit antioxidant activity and affect carbohydrate metabolism by delaying carbohydrate digestion, thus mitigating glucose in the circulation. Dairy products are effective matrices for delivering probiotics through the gastrointestinal tract. We compared the viability and antioxidant activity of the probiotics Lactobacillus acidophilus LA-5, Lacticaseibacillus rhamnosus GG, and Bifidobacterium animalis subsp. lactis in yogurt ice cream after in vitro digestion and compared α-amylase and α-glucosidase inhibition activities. Lacticaseibacillus rhamnosus GG had the highest viability after in vitro digestion (oral, gastric, and intestinal). Lactobacillus acidophilus LA-5 and Lacticaseibacillus rhamnosus GG exhibited the highest percentages of α-glucosidase (16.37% ± 0.32%) and α-amylase (41.37% ± 0.61%) inhibition. Bifidobacterium animalis subsp. lactis BB-12 and Lactobacillus acidophilus LA-5 showed the highest antioxidant activities via the α,α-diphenyl-β-picrylhydrazyl free radical-scavenging method and ferric-reducing antioxidant power assay, respectively. These findings suggest that yogurt ice cream can provide a suitable matrix for the delivery of probiotics from dairy culture to promote intestinal homeostasis with probiotic benefits in the host as well as a potential functional food to help reduce postprandial hyperglycaemia.

Antioxidant, antimicrobial, α-amylase and α-glucosidase inhibitory activities of methanolic leaf extract of Breynia vitis-idaea using in vitro and in silico techniques

Breynia vitis-idaea (Phyllanthaceae) is used in the Chinese traditional medicinal system, to treat chronic lung disease and wound healing. This study was aimed to identify the chemical constituents of B. vitis-idaea having biological properties by in-vitro and in-silico methods. Breynia vitis-idaea leaf extract was screened for the presence of phenol, flavonoid contents, antibacterial, antioxidant, and antidiabetic activities. Molecular docking of secondary metabolites with target proteins and ADMET analysis was done. Preliminary phytochemical screening of B. vitis-idaea contains a variety of phytoconstituents such as alkaloids, flavonoids, carbohydrates, tannin, saponin, terpenoids, phenol, protein, steroids and glycosides. The antibacterial activity of methanol leaf extract of B. vitis-idaea evaluated against Escherichia coli (17 mm), Klebsiella pneumoniae (16 mm), Bacillus subtilis (22 mm), and Staphylococcus aureus (18 mm) showed high activity. Antioxidant activities were assessed by ABTS and FRAP assays and their IC50 values were 41.06 µg/mL and 46.40 µL/mL, respectively. α-amylase and α-glucosidase activity showed IC50 values of 20.42 µg/mL and 14.60 µg/mL, respectively. Molecular docking study showed strong binding activity of γ-sitosterol to α-amylase and α-glucosidase, followed by guanosine and arbutin. ADMET analysis shows that γ-sitosterol, guanosine, and arbutin are good drug candidates. This work demonstrates the anti-diabetic potential of B. vitis-idaea.