Alpha-glucosidase Research Articles

Antioxidant and antidiabetic flavonoids from the leaves of Dypsis pembana (H.E.Moore) Beentje & J.Dransf., Arecaceae: in vitro and molecular docking studies

BackgroundOxidative stress and diabetes are medical conditions that have a growing prevalence worldwide, significantly impacting our bodies. Thus, it is essential to develop new natural antioxidant and antidiabetic agents. Dypsis pembana (H.E.Moore) Beentje & J.Dransf (DP) is an ornamental palm of the family Arecaceae. This study aimed to broaden the understanding of this plant’s biological properties by evaluating its in vitro antioxidant and antidiabetic activities.MethodsThe in vitro antioxidant activities of the crude extract, fractions, and selected isolates were evaluated by DPPH method. While the in vitro antidiabetic activities of these samples were evaluated by assessing the degree of inhibition of α-glucosidase. Additionally, molecular docking analysis was performed to investigate the interactions of tested compounds with two potential targets, the cytochrome c peroxidase and alpha glucosidase.ResultsThe crude extract displayed the highest antioxidant activity (IC50 of 11.56 µg/ml), whereas among the fractions, the EtOAc fraction was the most potent (IC50 of 14.20 µg/ml). Among tested compounds, isoquercetrin (10) demonstrated the highest potency, with an IC50 value of 3.30 µg/ml, followed by rutin (8) (IC50 of 3.61 µg/ml). Regarding antidiabetic activity, the EtOAc (IC50 of 60.4 µg/ml) and CH2Cl2 fractions (IC50 of 214.9 µg/ml) showed activity, while the other fractions did not demonstrate significant antidiabetic effects. Among tested compounds, kaempferol-3-O-neohesperidoside (9) showed the highest antidiabetic activity, with an IC50 value of 18.38 µg/ml, followed by kaempferol (4) (IC50 of 37.19 µg/ml). These experimental findings were further supported by molecular docking analysis, which revealed that isoquercetrin and kaempferol-3-O-neohesperidoside exhibited strong enzyme-binding affinities to the studied enzyme targets. This analysis provided insights into the structure-activity relationships among the investigated flavonol-O-glycosides.ConclusionThe biological and computational findings revealed that isoquercetrin and kaempferol-3-O-neohesperidoside have potential as lead compounds for inhibiting cytochrome c peroxidase and alpha glucosidase enzymes, respectively.

Exploring the interaction of phytochemicals from Hibiscus rosa-sinensis flowers with glucosidase and acetylcholinesterase: An integrated in vitro and in silico approach

Targeting multiple factors such as oxidative stress, alpha glucosidase and acetylcholinesterase (AChE) are considered advantageous for the treatment of diabetes and diabetes associated-cognitive dysfunction. In the present study, Hibiscus rosa-sinensis flowers anthocyanin-rich extract (HRA) was prepared. Phytochemical analysis of HRA using LC-ESI/MS/MS revealed the presence of various phenolic acids, flavonoids and anthocyanins. HRA showed in vitro antioxidant activity at low concentrations. HRA inhibited all the activities of mammalian glucosidases and AChE activity. The IC50 value of HRA for the inhibition of maltase, sucrase, isomaltase, glucoamylase and AChE was found to be 308.02 ± 34.25 µg/ml, 287.8 ± 19.49 µg/ml, 424.58 ± 34.75 µg/ml, 408.94 ± 64.82 µg/ml and 264.13 ± 30.84 µg/ml, respectively. Kinetic analysis revealed mixed-type inhibition against all the activities except for glucoamylase (competitive) activity. In silico analysis confirmed the interaction of two active constituents cyanidin 3-sophoroside (CS) and quercetin 3-O-sophoroside (QS) with four subunits, n-terminal and c-terminal subunits of human maltase-glucoamylase and sucrase-isomaltase as well as with AChE. Molecular dynamics simulation, binding free energy calculation, DCCM, PCA, PCA-based free energy surface analysis ascertained the stable binding of CS and QS with target proteins studied. HRA could be used as complementary therapy for diabetes and cognitive improvement.

Identification of antidiabetic inhibitors from Allophylus villosus and Mycetia sinensis by targeting α-glucosidase and PPAR-γ: In-vitro, in-vivo, and computational evidence

Diabetes mellitus (DM) is a metabolic disorder arising from insulin deficiency and defectiveness of the insulin receptor functioning on transcription factor where the body loses control to regulate glucose metabolism in β-cells, pancreatic and liver tissues to homeostat glucose level. Mainstream medicines used for DM are incapable of restoring normal glucose homeostasis and have side effects where medicinal plant-derived medicine administrations have been claimed to cure diabetes or at least alleviate the significant symptoms and progression of the disease by the traditional practitioners. This study focused on screening phytocompounds and their pharmacological effects on anti-hyperglycemia on Swiss Albino mice of n-hexane, ethyl acetate, and ethanol extract of both plants Mycetia sinensis and Allophylus villosus as well as the in-silico investigations. Qualitative screening of phytochemicals and total phenolic and flavonoid content estimation were performed significantly in vitro analysis. FTIR and GC–MS analysis précised the functional groups and phytochemical investigations where FTIR scanned 14, 23 & 17 peaks in n-hexane, ethyl acetate, and ethanol extracts of Mycetia sinensis whereas the n-hexane, ethyl acetate, and ethanol extracts of Allophylus villosus scanned 11 peaks, 18 peaks, and 29 peaks, respectively. In GC–MS, 24 chemicals were identified in Mycetia sinensis extracts, whereas 19 were identified in Allophylus villosus extracts. Moreover, both plants' ethyl acetate and ethanol fractioned extracts were reported significantly (p < 0.05) with concentrations of 250 mg and 500 mg on mice for oral glucose tolerance test, serum creatinine test and serum alkaline phosphatase test. In In silico study, a molecular docking study was done on these 43 phytocompounds identified from Mycetia sinensis and Allophylus villosus to identify their binding affinity to the target Alpha Glucosidase (AG) and Peroxisome proliferator-activated receptor gamma protein (PPARG). Therefore, ADMET (absorption, distribution, metabolism, excretion, and toxicity) analysis, quantum mechanics-based DFT (density-functional theory), and molecular dynamics simulation were done to assess the effectiveness of the selected phytocompounds. According to the results, phytocompounds such as 2,4-Dit-butyl phenyl 5-hydroxypentanoate and Diazo acetic acid (1S,2S,5R)-2-isopropyl-5-methylcyclohexyl obtained from Mycetia sinensis and Allophylus villosus extract possess excellent antidiabetic activities.

Phytochemical profiling and antidiabetic evaluation of Peperomia pellucida as a potential alpha glucosidase inhibitor

Abstract. Hidayati S, Agustin AT, Sari EK, Sari SM, Destiawan RA, Silvana WA. 2023. Phytochemical profiling and antidiabetic evaluation of Peperomia pellucida as a potential alpha glucosidase inhibitor. Biodiversitas 24: 5972-5978. One strategy in post-prandial hyperglycemic control can be done by inhibiting the digestion of dietary carbohydrates through the inhibition of alpha glucosidase enzymes. This study was conducted to determine the antidiabetic activity of Peperomia pellucida in inhibiting the alpha glucosidase. An inhibitory activity analysis test was carried out on alpha glucosidase in vitro and in silico using alpha glucosidase. Molecular docking between receptors and ligands was performed using Hex 8.0 software. The setting column is set in Shape+Electro+DARS mode. The results showed ethanol extract and ethyl acetate fraction in vitro showed the ability to inhibit the activity of alpha glucosidase enzyme with C50 values of 13.43 mg/mL and 9.73 mg/mL respectively with IC50 positive control acarbose values of 8.11 mg/mL. The results of in silico analysis showed that the Patuloside A component was able to bind to the binding site of alpha glucosidase and gave the smallest binding energy value with a value of -321.4 kcal/mol compared to isovitexin, isoswertisin, pellucidatin, and caryatin-7-O-?-rhamnoside compounds. P. pellucida has the potential to be developed as an antidiabetic agent that has activity in inhibiting the work of the enzyme alpha glucosidase so that it can reduce blood glucose levels.

Investigating the viral ecology and contribution to the microbial ecology in full-scale mesophilic anaerobic digesters

In an attempt to assess the diversity of viruses and their potential to modulate the metabolism of functional microorganisms in anaerobic digesters, we collected digestate from three mesophilic anaerobic digesters in full-scale wastewater treatment plants treating real municipal wastewater. The reads were analyzed using bioinformatics algorithms to elucidate viral diversity, identify their potential role in modulating the metabolism of functional microorganisms, and provide essential genomic information for the potential use of virus-mediated treatment in controlling the anaerobic digester microbiome. We found that Siphoviridae was the dominant family in mesophilic anaerobic digesters, followed by Myoviridae and Podoviridae. Lysogeny was prevalent in mesophilic anaerobic digesters as the majority of metagenome-assembled genomes contained at least one viral genome within them. One virus within the genome of an acetoclastic methanogen (Methanothrix soehngenii) was observed with a gene (fwdE) acquired via lateral transfer from hydrogenotrophic methanogens. The virus-mediated acquisition of fwdE gene enables possibility of mixotrophic methanogenesis in Methanothrix soehngenii. This evidence highlighted that lysogeny provides fitness advantage to methanogens in anaerobic digesters by adding flexibility to changing substrates. Similarly, we found auxiliary metabolic genes, such as cellulase and alpha glucosidase, of bacterial origin responsible for sludge hydrolysis in viruses. Additionally, we discovered novel viral genomes and provided genomic information on viruses infecting acidogenic, acetogenic, and pathogenic bacteria that can potentially be used for virus-mediated treatment to deal with the souring problem in anaerobic digesters and remove pathogens from biosolids before land application. Collectively, our study provides a genome-level understanding of virome in conjunction with the microbiome in anaerobic digesters that can be used to optimize the anaerobic digestion process for efficient biogas generation.

Anthocyanins of Hierbamora (Solanum nigrescens): Revealing their Nutraceutical Potential for Controlling Hypertriglyceridemia and Helicobacter pylori Viability.

We present the inhibitory properties of the Solanum nigrescens anthocyanin fraction (SNAF) and its major constituents on alpha-glucosidase (AG), pancreatic lipase (PL), HMG-CoA reductase, and ornithine decarboxylase (ODC). The effect of SNAF was simultaneously evaluated in ICR male mice exposed to triglyceride charge test (TCT). HPLC-MS profiling revealed the presence cyanidin-3-O-rutinoside-5-glucoside (CRG), delphinidin-3-(p-coumaroyl)-rutinoside-5-glucoside (DCRG), and petunidin-3-(cis-p-coumaroyl)-rutinoside-5-glucoside (PCRG) as major constituents of the fraction. SNAF, CRG, and specially PCRG, induced strong non-competitive inhibition on PL (IC50 , 33-86 μg mL-1 ). The results of TCT confirmed their capacity to ameliorate (p <0.001) hypertriglyceridemia during postprandial and interdigestive stages. SNAF, CRG, DCRG, and PCRG caused negligible growth inhibition (MIC>600 μg mL-1 ) on beneficial bacteria whereas SNAF and DCRG exerted inhibitory activity on Helicobacter pylori ATCC 53504 (MIC,187-64 μg mL-1 ). Additional exploration revealed that SNAF and DCRG produced non-competitive activity on H. pylori urease, which facilitates bacterial growth under acidic conditions.

Wound healing and antidiabetic properties of green synthesized silver nanoparticles in 3T3-L1 mouse embryo fibroblast cells through 2-NBDG expression

The antibacterial and antifungal properties of silver and silver-based compounds have been recognized for quite some time. This study employed an extracellular method to produce silver nanoparticles (AgNPs) using the leaves of Barringtonia racemosa (B. racemosa). B. racemosa's aqueous leaf extract underwent a color change from grey to dark brown, which revealed the formation of AgNPs. UV spectrophotometry analysis confirmed the presence of silver with the maximum absorbance of 427 nm. FTIR analysis supported the existence of alcohols with the OH stretch and alkenes with the CC stretch. The existence of AgNPs with an average hydrodynamic diameter of 36.58 nm was confirmed using dynamic light scattering (DLS). Energy dispersive X-ray spectroscopy (EDX) confirmed the presence of silver. Additionally, the AgNPs showed spherical structure particles with the size distribution ranging from 10 to 17 nm with polydispersity, under transmission electron microscopy (TEM). Thermogravimetric analysis (TGA) findings revealed that AgNPs maintain good thermal stability even at high temperatures. A variety of bacterial and fungal pathogens were successfully inhibited by the AgNPs, with S. epidermidis (27 mm) showing the strongest antibacterial activity and a 20 mm inhibitory zone was against Candida albicans for its antifungal activity. Furthermore, AgNPs also showed a positive synergistic impact with the tested antibiotic and antifungal agents. In addition, the AgNPs demonstrated strong antioxidant activity. The considerable inhibition of alpha amylase and alpha glucosidase by the AgNPs were noted by their respective IC50 values of 16.65 μg/ml and 18.97 μg/ml. Flow cytometry analysis revealed a significant increase in glucose uptake by 3T3-L1 cells after exposure to AgNPs, as indicated by the upregulation of 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino)-2-deoxyglucose (2-NBDG) expression. In 3T3/L cells, the AgNPs also demonstrated remarkable potential in accelerating wound healing achieving 92% closure of wounds after 48 h of incubation. As a result, it can be concluded that the AgNPs exhibit positive antidiabetic properties, suggesting their potential applications in the pharmaceutical industry including in the management of diabetic foot ulcers.

Chalcones: The flavonoid derivatives synthesis, characterization, their antioxidant and in vitro/in vivo antidiabetic potentials

Chalcones (designated JA1, JA2 and JA3) were prepared from aromatic aldehyde and acetophenone which were then characterized using various spectroscopic techniques. The antioxidant potential of synthesized compounds was evaluated against DPPH free radical whereas the antidiabetic potential was determined against alpha glucosidase. Further the antidiabetic potential of the synthesized compounds was evaluated in rat model which were given orally experimental animals in doses 10 and 20 mg/kg body weight. The blood biochemical parameters like total cholesterol, triglycerides, alanine phosphatase, serum glutamic pyruvic transaminase, serum glutamic oxaloacetic transaminase, serum creatinine, HDL, and LDL levels were determined using commercially available kits. The antioxidant potential was found high for JA3 followed by JA2 with IC50 value of 64.02 ± 1.47 μg/ml whereas against alpha glucosidase again the same compound with IC50 of 63.04 μg/ml exhibited highest inhibitory potential. The blood glucose level was brought to almost normal level (126.88 and 119.13 mg/dl at 10 and 20 mg/kg body weight) in diabetic rats (induced by STZ) by compound JA3 at the tested doses in comparison to acarbose at day 28th. The blood biochemical parameters were normalized in diabetic rats by compound JA3 compared with diabetic control group. Based on the results JA3 should be considered as effective antioxidant and antidiabetic drug candidate.

Effect of Phenolics from Aeonium arboreum on Alpha Glucosidase, Pancreatic Lipase, and Oxidative Stress; a Bio-Guided Approach.

Metabolic syndrome (MetS) is a global issue affecting over a billion people, raising the risk of diabetes, cardiovascular disorders, and other ailments. It is often characterized by hypertension, dyslipidemia and/or obesity, and hyperglycemia. Chemical investigation of Aeonium arboreum (L.) Webb & Berthel led to the isolation of six compounds, viz. β-sitosterol, β-sitosterol glucoside, myricetin galactoside, quercetin rhamnoside, kaempferol rhamnoside, and myricetin glucoside. Interestingly, A. arboreum's dichloromethane (DCM), 100 and 50% MeOH Diaion fractions and the isolated compound (quercetin-3-rhamnoside) revealed potent α-glucosidase inhibitory activity, especially 50% Diaion fraction. In addition, they also showed very potent antioxidant potential, especially the polar fractions, using DPPH, ABTS, FRAP, ORAC, and metal chelation assays. Notably, the 50% Diaion fraction had the highest antioxidant potential using DPPH and ORAC assays, while the 100% Diaion fraction and quercetin-3-rhamnoside showed the highest activity using ABTS, FRAP, and metal chelation assays. Also, quercetin-3-rhamnoside showed a good docking score of -5.82 kcal/mol in comparison to acarbose. In addition, molecular dynamic stimulation studies illustrated high stability of compound binding to pocket of protein. Such potent activities present A. arboreum as a complementary safe approach for the management of diabetes mellitus as well as MetS.

ISOLATION, CHARACTERIZATION OF Β-SITOSTEROL FROM THE HERBAL EXTRACTS AND EVALUATION OF IN VITRO ANTI-OXIDANT AND ANTI-DIABETIC ACTIVITIES

Plants naturally produce phytochemicals, physiologically active chemical compounds that serve as a natural defence mechanism and have considerable potential for treating a wide range of ailments. Cyperus rotundus and Tinospora cordifolia are the potent traditional herbs used in Ayurveda medicine, especially for diabetes. The current study aimed to isolate and characterize the bioactive compound β-sitosterol from the combined ethanolic extracts of the rhizome of Cyperus rotundus (C. rotundus) and the stem of Tinospora cordifolia (T. cordifolia). Furthermore, the in vitro antioxidant and anti-diabetic activities of isolated compounds are tested by DPPH assay, alpha amylase inhibition assay, alpha glucosidase inhibition assay and glucose uptake activity in 3T3L1 cells. The combined ethanolic extract of both medicinal plants was initially subjected to column and preparative thin layer chromatography for the isolation of bioactive compounds, followed by their characterization by spectral methods like UV, FT-IR, 1H and 13C NMR. The structure of the isolated compound is identified as β-sitosterol and further confirmed by structural elucidation. Similarly, the isolated compound β-sitosterol exhibited higher antioxidant scavenging activity of 64.54±1.38 µg/ml against DPPH at a concentration 100µg/ml. The study also confirmed that β-sitosterol showed good inhibitory activities towards alpha-amylase (62.21±1.25 µg/ml) and alpha-glucosidase (65.48±1.47 µg/ml). β-sitosterol showed significantly increased glucose uptake (28% at 100μg/ml). Based on the conclusion that the organic compound β-sitosterol exhibited good antioxidant and anti-diabetic efficacy, it could have been used in the treatment of free radical disease and can be utilized in the management of diabetes mellitus.

In vitro study of antioxidant, antigylycation, sugar hydrolysis enzyme inhibitory effect and molecular in silico docking study of angularly condensed diquinothiazines

BackgroundAngularly condensed diquinothiazines (MY1, MY2, and MY3) a phenothiazine derivative was evaluated for antioxidant, antiglycation, sugar hydrolysis enzyme inhibitory effect and molecular in silico docking. MethodAntioxidants were evaluated using DPPH, chelating ion, FRAP and lipid peroxidation. Alpha glucosidase and alpha amylase inhibition was used for sugar hydrolysis enzyme inhibitory activity and antiglycation with BSA-Glucose and BSA-MGO. In silco docking was done with AutoDock Vina and ADMET properties with pkCSM software. ResultAngularly condensed diquinothiazines (MY1, MY2, and MY3) showed free radical scavenging, antiglycation and alpha-glucosidase and alpha-amylase inhibition. Insilco docking and pkCSM ADME shows interaction with aldose reductase, glyoxalase 1, receptor AGE, alpha-glucosidase, and alpha-amylase. ConclusionAntiglycation, alpha-glucosidase, and alpha-amylase inhibition activity of Angularly condensed diquinothiazines maybe through deactivation of aldose reductase enzyme activity, activation of glyoxalase 1 pathway, reduction of dicarbonyl stress and oxidative stress.

Two fluorimetric determinations of acid α-glucosidase activity in dried blood spot: Pompe disease in Iranian population

IntroductionPompe disease is a lysosomal storage disorder. This study aimed to validate and compare 2 fluorimetric methods for measuring α-glucosidase acid activity in dried blood spot sample (DBS), with potential applications in neonatal screening, and disease follow-up of Pompe patients among the Iranian population for the first time. Materials and methodsThe evaluation involved 3 enzyme levels and 7 parameters. The analysis included 141 Healthy individuals, 8 Pompe patients, and 10 obligate heterozygotes using reference and modified methods. ResultsBoth methods exhibited highly linear calibration curves. The limit of detection (LOD) and limit of quantification (LOQ) were obtained in the micromolar concentration range in 2 methods. Inter-day and intra-day precision, expressed as relative standard deviations (RSD%) were calculated. The normal ranges were determined in healthy individuals. Receiver operating characteristic (ROC) curves were analyzed, and 2 parameters, total neutral α-glucosidase (NAG)/acid α-glucosidase (GAA) and pH ratio, were identified as cut-off values with excellent accuracy, sensitivity, and specificity for evaluating Pompe disease in both methods. ConclusionsEstablishing and implementing these 2 methods for the Iranian population effectively differentiated between healthy and patient individuals. Method II, with its shorter incubation time, demonstrated practicality in the clinical setting.

Synthesis of camphor thiazole derivates from Dryobalanops aromatica and its bioactivity as antioxidants and antidiabetes against alpha glucosidase enzymes

Camphor is synthesized from the Sumatran camphor plant (Dryobalanops aromatica) in previous experiments. It can be synthesized with thiosemicarbazide, ethy-2‑chloro acetoacetate, and sodium acetate (catalyze) to form camphor derivate with thiazole ring structure. Hydrazine and phenylhydrazine were both used to make the thiazole ring variations. All the compounds were purified by recrystallization method and characterized by TLC, FTIR, UV–vis, and LC-MS. Camphor thiazole (Product 1), camphor thiazole hydrazine (Product 2), and camphor thiazole phenylhydrazine (Product 3) were successfully synthesized with%yields of 73.24 %; 77.36 %; and 72.91 % respectively. Furthermore, their antioxidant activity was measured using the DPPH free radical method. Product 2 had the strongest antioxidant activity with IC50 value of 6.93 ppm. The antidiabetic activity was measured using the α-glucosidase enzyme. This indicated that product 1 had the best inhibitory activity against the α-glucosidase enzyme with IC50 values of 869.06 ppm.•We developed an alternative method to utilize camphor extracted from the D. aromatica plant to be used as an alternative medicinal ingredient related to antioxidants and antidiabetes.•All products were successfully synthesized and have the potential to be used as antioxidants with an IC50 value of 6.93 ppm for Product 2 and as antidiabetics by means of an α-glucosidase inhibitor with an IC50 value of 869.06 ppm for Product 1.

Phytochemical profiling, human insulin stability and alpha glucosidase inhibition of Gymnema latifolium leaves aqueous extract: Exploring through experimental and in silico approach

Diabetes mellitus Type 2 (DM2T) is a rapidly expanding metabolic endocrine disorder worldwide. It is caused due to inadequate insulin secretion by pancreatic beta cells as well as development of insulin resistance. This study aimed to investigate the anti-α-glucosidase, insulin stabilization effect, and non-cytotoxic nature of Gymnema latifolium leaf aqueous extract (GLAE). FTIR analysis revealed the functional groups of compounds present in GLAE. Through LC/ESI-MS/MS analysis, about 12 compounds which belongs to different classes, triterpene glycosides, flavonoids, phenolics, stilbene glycosides and chlorophenolic glycosides were identified. GLAE showed in vitro antioxidant activity. GLAE stabilized insulin by increasing its α-helical content. GLAE inhibited the mammalian α-glucosidase (IC50 = 144 μg/mL) activity through competitive mode (Ki = 61.30 µg/mL). GLAE did not affect the viability of normal cell line (Vero cell line) which shows its non-toxic nature. Molecular docking of phytocompounds identified in GLAE was done with human α-glucosidase and insulin. The top 2 compounds [Gymnema saponin V (GSV) and quercetin 3-(2-galloylglucoside) (QGG) with α-glucosidase; GSV and Z)-resveratrol 3,4′-diglucoside (RDG) with human insulin] with low binding free energy were subjected to 100 ns molecular dynamics simulation to ascertain the stable binding of ligand with protein. The MM/GBSA analysis revealed binding free energy of GSV/α-glucosidase and QGG /α-glucosidase to be - 20.9935 and, − 30.9461 kcal/mol, respectively. Altogether GLAE is valuable source of anti-α-glucosidase inhibitors and insulin stabilizing compounds, suggesting potential lead for further exploration as complementary medicine against DM2T.

Validating the Nutraceutical Significance of Minor Millets by Employing Nutritional-Antinutritional Profiling.

Millets are group of underutilized cereal crops with higher nutritional values. The present investigation used different classes of minor millets, including barnyard (sava), little (kutki), finger (ragi), kodo and foxtail millets, for evaluation of their nutritional parameters, i.e., the content of proteins, total amino acids, total sugars, insoluble fibers, soluble fibers, total dietary fibers, iron (Fe) and zinc (Zn), along with antinutritional and antioxidant parameters, viz., tannic acid, phytic acid, phenol, flavonoid, proline and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity. Alpha amylase and alpha glucosidase activity were also thought to elevate millets as a viable staple meal. Foxtail millet showed the maximum inhibition, with an IC50 value of 20.46 ± 1.80 µg mL-1 with respect to α-amylase. The coefficient of correlation between nutritional and antinutritional compositions showed that the starch content was significantly and positively correlated with insoluble fiber (r = 0.465) and dietary fiber (r = 0.487). Moreover, sugar was positively correlated with the phytic acid (r = 0.707), Fe and Zn (r = 0.681) contents. To determine the peptides responsible for anticancer activity, the foxtail protein was subjected to ultrafiltration; it was found that the 3 kDa fraction retained the greatest anticancer activity. Selected millet germplasm line(s) that have the best nutraceutical properties could be used in millet improvement programs.

Pharmacokinetics of Alglucosidase Alfa Manufactured at the 4000-L Scale in Participants with Pompe Disease: A Phase 3/4 Open-Label Study.

Pompe disease is a rare, autosomal recessive, degenerative neuromuscular disease caused by deficiency of acid α-glucosidase, a lysosomal enzyme that degrades α-1,4 and α-1,6 linkages in glycogen. The objectives of this study (PAPAYA; NCT01410890) were to (1) characterize the pharmacokinetics of 20mg/kg body weight alglucosidase alfa manufactured at the 4000-L scale following a single intravenous dose in participants aged less than 18 and 18 years or older with Pompe disease and (2) evaluate the relationship between anti-alglucosidase alfa antibody titers and the pharmacokinetics of alglucosidase alfa. Mean maximum plasma concentration and area under the concentration-time curve from time zero and extrapolated to infinite time were 204μg/mL and 1110μg•h/mL for participants aged less than 18 years (n = 10), respectively, and 307μg/mL and 1890μg•h/mL for participants aged 18 years or older (n = 10), respectively. Mean terminal half-life was 5.43 hours in participants aged less than 18 years with a high variability (70%) and 3.84 hours in participants aged 18 years or older with a low variability (21%). Mean maximum plasma concentration and area under the concentration-time curve from time zero and extrapolated to infinite time were 256μg/mL and 1452μg •h/mL, respectively, in anti-alglucosidase alfa-negative participants (n = 12) and 262μg/mL and 1703μg•h/mL, respectively, in anti-alglucosidase alfa-positive participants (n = 7). The study findings enrich available data from existing information on alglucosidase alfa without changing its known risks and benefits.

Nutraceutical Activity of Anthocyanins from the Edible Berries of Rhamnus pompana.

We present the inhibitory properties of the R. pompana anthocyanin fraction (RPAF) and its major constituents on alpha-glucosidase (AG), pancreatic lipase (PL), HMG-CoA reductase, and ornithine decarboxylase (ODC). The effect of RPAF was also evaluated in ICR male mice subjected to oral glucose tolerance test (OGTT) and hypercaloric/atherogenic diet for 30 days. RP-HPLC/MS profiling revealed that RPAF contained five major anthocyanins and induced slight inhibition on PL and HMG-CoA reductase (IC50 , 245-338 μg mL-1 ) whereas strong activity on AG and ODC (IC50 , 130-133 μg mL-1 ) was observed. Kinetic studies and molecular docking with pelargonidin-3-O-rutinoside (P3R) on ODC, revealed changes in Km (0.9514-0.9746 mM) and Vmax (1.96-2.32 μmol mg-1 min-1 ) suggesting mixed inhibition and molecular interaction with two active sites of ODC. P3R showed antiproliferative activity (IC50, 46.5 μM) and decreased polyamine accumulation in DLD-1 cells. The results of OGTT confirmed that RPAF regulates postprandial glucose levels in diabetic animals which experienced a significant glucose depletion (30 %; p<0.001) from 30 to 120 min post-treatment. Prolonged supplementation of RPAF caused significant decrease (p<0.001) in plasma glucose, total cholesterol, LDL-c and triglycerides as well as significant increase (p<0.001) of HDL-c compared with normoglycemic untreated animals.

Pharmacognostical and Pharmacological Evaluation of leaves of Citrullus lanatus L.

Citrullus lanatus (Watermelon) is commonly available throughout India and traditionally used in treatment of various ailments. This plant is well-explored for its fruit but the potential of its leaves is still unexplored. Thus, the present study focuses on pharmacognostical and pharmacological evaluation of leaves of Citrullus lanatus. In pharmacognostical evaluation, all macroscopic, microscopic, qualitative, quantitative and microbiological analysis were performed. In pharmacological evaluation, the study mainly focuses on the potential of leaves for antioxidant and antidiabetic activities. The antioxidant activity has been performed by various methods such as DPPH and phospho-molybdenum method. The antidiabetic activity has been performed by non-enzymatic glycosylation of hemoglobin assay method, glucose uptake in yeast cells method, alpha amylase inhibition assay method and alpha glucosidase inhibition assay method. The results of antioxidant activity shows that the plant extract possess good free radical scavenging properties due to the presence of vitamin C, polyphenol and flavonoid content. The results of antidiabetic activity by non-enzymatic glycosylation method shows that the plant extract of C. lanatus exhibit significant inhibition of glycosylation as compared with standard drug alpha tocopherol. In glucose uptake by yeast cells method, the extract showed greater efficiency in increasing the glucose uptake by yeast cells as compared to standard drug acarbose. In alpha amylase inhibition method, Alpha amylase inhibitors bind to alpha bond of polysaccharide and prevent breakdown of polysaccharide in mono and disaccharide which shows significant activity of plant extract as compared to an acarbose standard drug. Thus from the above findings, it can be concluded that C. lanatus leaves possess a great potential for antioxidant and antidiabetic activity and this plant should be more researched and formulation should be marketed in the recent future.&#x0D; Keywords: Citrullus lanatus (Cucurbitaceous), Watermelon, Anti-oxidant Activity, Anti-diabetic Activity

Study of Optimal Conditions to Grow Thai Ganoderma, Fruiting Test, Proximate and Their Alpha Glucosidase Inhibitory Activity.

Ganoderma (Ganodermataceae) has a worldwide distribution and has been widely used in traditional medicines. In this study, we report wild strains of Ganoderma that include two G. sichuanense and one G. orbiforme from northern Thailand. Optimal conditions for mycelium growth were ensured. The most favourable medium was potato sucrose agar for G. sichuanense and oatmeal agar for G. orbiforme and at 25 °C and 30 °C and pH 4-8. All types of cereal grains can be used to promote the growth of the mycelia of Ganoderma species. Fruiting tests were performed. All strains of Ganoderma produce fruiting bodies successfully in bag culture at 28 ± 1 °C with 75-85% relative humidity. Only G. orbiforme produced fruiting bodies in field cultivation at the laboratory scale. In the first flush yields, the G. sichuanense strain MFLUCC 22-0064 gave better production (the B.E was 152.35 ± 6.98 g). This study is the first to document the bag and field cultivation of wild Thai G. orbiforme. Ganoderma species are revealed to contain high amounts of fiber (47.90-52.45% d.b.), protein (12.80-14.67% d.b.), fat (4.90-5.70% d.b.), and carbohydrates (3.16-4.02% d.b.). Additionally, G. sichuanense and G. orbiforme were preliminarily screened for biological activity for inhibition of alpha-glucosidase enzyme activity. The IC50 values of G. orbiforme (MFLUCC 22-0066) was 105.97 ± 1.36 µg/mL and G. sichuanense (MFLUCC 22-0064) was 126.94 ± 0.87 µg/mL. Both strains had better inhibition than acarbose (168.18 ± 0.89 µM). These results on wild strains of Ganoderma will be useful for further studies on the applications of Ganoderma. Later the species can be introduced to domestic markets for cultivation and medicinal use.

Increasing Enzyme Mannose-6-Phosphate Levels but Not Miglustat Coadministration Enhances the Efficacy of Enzyme Replacement Therapy in Pompe Mice.

Pompe disease is a rare glycogen storage disorder caused by a deficiency in the lysosomal enzyme acid α-glucosidase, which leads to muscle weakness, cardiac and respiratory failure, and early mortality. Alglucosidase alfa, a recombinant human acid α-glucosidase, was the first approved treatment of Pompe disease, but its uptake into skeletal muscle via the cation-independent mannose-6-phosphate (M6P) receptor (CIMPR) is limited. Avalglucosidase alfa has received marketing authorization in several countries for infantile-onset and/or late-onset Pompe disease. This recently approved enzyme replacement therapy (ERT) was glycoengineered to maximize CIMPR binding through high-affinity interactions with ∼7 bis-M6P moieties. Recently, small molecules like the glucosylceramide synthase inhibitor miglustat were reported to increase the stability of recombinant human acid α-glucosidase, and it was suggested that an increased serum half-life would result in better glycogen clearance. Here, the effects of miglustat on alglucosidase alfa and avalglucosidase alfa stability, activity, and efficacy in Pompe mice were evaluated. Although miglustat increased the stability of both enzymes in fluorescent protein thermal shift assays and when incubated in neutral pH buffer over time, it reduced their enzymatic activity by ∼50%. Improvement in tissue glycogen clearance and transcriptional dysregulation in Pompe mice correlated with M6P levels but not with miglustat coadministration. These results further substantiate the crucial role of CIMPR binding in lysosomal targeting of ERTs. SIGNIFICANCE STATEMENT: This work describes important new insights into the treatment of Pompe disease using currently approved enzyme replacement therapies (ERTs) coadministered with miglustat. Although miglustat increased the stability of ERTs in vitro, there was no positive impact to glycogen clearance and transcriptional correction in Pompe mice. However, increasing mannose-6-phosphate levels resulted in increased cell uptake in vitro and increased glycogen clearance and transcriptional correction in Pompe mice, further underscoring the crucial role of cation-independent mannose-6-phosphate receptor-mediated lysosomal targeting for ERTs.