Identification of phytocompounds as potent inhibitors of sodium/glucose cotransporter-2 leading to diabetes treatment.

Type-II diabetes, a major metabolic disorder has threatened the very existence of a healthy life since long ago. Commercially available antidiabetic drugs are known for several adverse effects. The present study attempted to identify potential phytocompounds as inhibitors of sodium/glucose cotransporter-2 (SGLT2), a major protein that helps in glucose re-absorption from renal tubules. A total of 28 phytocompounds were collected based on the literature survey. 3D co-ordinates of phytocompounds were collected from PubChem database. Molecular docking was carried out with SGLT2 protein and the best 3 docking complexes were subjected to molecular dynamics simulation for 100 ns. Free energy changes were also analyzed using MM/PBSA analysis. Phytocompounds were also analyzed for their drug-likeness and ADMET properties. Docking study observed a strong binding affinity of phytocompounds (> -7.0 kcal/mol). More than 10 phytocompounds showed better binding affinity compared to reference drugs. Further analysis of three best docking complexes when analyzed by MD simulation showed better stability and compactness of the complexes compared to reference drug, empagliflozin. MM/PBSA analysis also revealed that van der Waals force and electrostatic energy are the major binding energy involved in the complex formation. Like docking energy, free energy analysis also observed stronger binding energies (ΔGGAS) in SGLT2-phytocompound complexes compared to empagliflozin complex. All the phytocompounds showed drug-likeness and considerable ADMET properties. The study, therefore, suggests that Trifolirhizin-6'-monoacetate, Aspalathin, and Quercetin-3-glucoside could be a possible inhibitor of SGLT2 protein. However, further studies need to be carried out to reveal the exact mode of activity.Communicated by Ramaswamy H. Sarma.

Impact assessment of Intensified Malaria Control Project in transitioning a high malaria-endemic district to a low-endemic district: an epidemiological aspect.

Intensified Malaria Control Project (IMCP) was implemented in 2005 to control malaria in all North-Eastern and Odisha states of India. The present study aimed to investigate the impact of IMCP in reducing the malaria burden in Udalguri district, Assam state of North-East India. Malaria epidemiological data were obtained for IMCP intervention (Udalguri) and nonintervention district (West Singhbhumi, Jharkhand state). IMCP activities include introducing bi-valent rapid diagnostic kits (RDTs), Artemether-Lumefantrine drug in North-East India, long-lasting insecticidal nets (LLINs) distribution, and creating awareness programs about malaria in an intensified mode. The data revealed a significant decline in annual parasite incidence (API) from 14.94 (2005) to 2.61 (2018), -37% (95%CI: -57%, -19%, p = 001) after using LLINs in 2009 and -64% (95%CI: -116%, -14%, p = 013) after the introduction of RDTs in district Udalguri. Whereas control district showed a -28% (95%CI: -63%, 6.3%, p = 0.051) decrease in API using LLINs and a 10% (95%CI: -7.6%, 28%, p = 0.122) increase after the introduction of RDTs. Plasmodium falciparum (Pf) and P. vivax (Pv) were the major malarial parasites in Udalguri. Pv-malaria was much higher (71%) than Pf-malaria (29%) during the study period. An increasing trend of Pf cases was observed in Udalguri. Udalguri and Khoirabari BPHCs showed an overall reduction of 94% (95%CI: -143%, -45%, p = 0.001) and 84% (95%CI: -126%, -39%, p = 0.003), respectively; however, only a 10% (95%CI: -65%, -41%, p = 0.360) reduction in API was observed in Orang BPHC. An overall decrease in malaria indicates the effective implementation of vector and disease control strategies in the Udalguri district.

Phenolic compounds as α-glucosidase inhibitors: a docking and molecular dynamics simulation study.

Phenolic compounds possess significant biological activity. Several pieces of research emphasize the medicinal importance of phenolic compounds, including diabetes. The present study investigated the α-glucosidase inhibitory activity of phenolic compounds reported from several plants. The phenolic compounds reported in different literature were collected. Molecular docking was carried out using AutoDock Vina. Various physicochemical properties such as size, LogP, molecular complexity, hydrogen bonding properties of phenolic compounds were correlated with the binding affinities. Furthermore, MD simulation was carried out to study the structural stability of the docking complexes. A total of 155 phenolic compounds were reported from different plants. Amentoflavone showed the strongest binding affinity with α-glucosidase, much more potent than reference acarbose. The binding energy showed a good correlation with the molecular complexity, hydrogen bond donor and acceptor property and heavy atom counts of the compounds. The polarity of the surface area also showed a positive correlation with the binding affinity of the compounds. The best docking phenolic compound, amentoflavone, showed stable binding affinity and conformation during the simulation period compared to apoprotein and acarbose-docking complex. The top ten phenolic compounds, including amentoflavone, showed considerable drug-likeness properties with fewer toxicity effects. The study suggests that the amentoflavone could be a potential therapeutic drug as an α-glucosidase inhibitor and help control postprandial hyperglycemia.Communicated by Ramaswamy H. Sarma.

Moderate Rainfall and High Humidity During the Monsoon Season, Negligence in Using Malaria Protection Methods and High Proportion of Mild Symptomatic Patients Were the Driving Forces for Upsurge of Malaria Cases in 2018 Among Tea Tribe Populations in Endemic Dolonibasti Health Sub-center, Udalguri District, Assam State, North-East India.

Malaria elimination is a global priority, which India has also adopted as a target. Despite the malaria control efforts like long-lasting insecticidal nets distribution, rounds of indoor residual spray, the introduction of bi-valent rapid diagnostic tests and artemisinin combination therapy, malaria remained consistent in Dolonibasti sub-center of Orang block primary health center (BPHC) under the district Udalguri, Assam state followed by abrupt rise in cases in 2018. Therefore, we aimed to investigate the factors driving the malaria transmission in the outbreak area of Dolonibasti sub-center. Malaria epidemiological data (2008-2018) of Udalguri district and Orang BPHC was collected. The annual (2011-2018) and monthly (2013-2018) malaria and meteorological data of Dolonibasti sub-center was collected. An entomological survey, Knowledge, Attitude and Practices study among malaria cases (n = 120) from Dolonibasti was conducted. In 2018, 26.1 % (2136/ 8188) of the population of Dolonibasti were found to be malaria positive, of which 55% were adults (n = 1176). Majority of cases were from tea tribe populations (90%), either asymptomatic or with fever only, 67.5 % (81/120) had experienced malaria infection during past years. The outbreak was characterized by a strong increase in cases in June 2018, high proportion of slide falciparum rate of 26.1% (other years average, 15.8%) and high proportion of P. falciparum of 81.2 % (other years average, 84.3%). Anopheles minimus s.l. was the major vector with 28.6% positivity and high larval density in paddy fields/ drainage area. Annual relative humidity was associated with rise in malaria cases, annual parasite incidence (rs = 0.69, 90%CI; p = 0.06) and slide positivity rate (rs = 0.83, 95%CI; p = 0.01). Older people were less educated (rs = -0.66; p < 0.001), had lesser knowledge about malaria cause (rs = -0.42; χ2=21.80; p < 0.001) and prevention (rs = -0.18; p = 0.04). Malaria control practices were followed by those having knowledge about cause of malaria (rs = 0.36; χ2 = 13.50; p < 0.001) and prevention (rs = 0.40; χ2 = 17.71; p < 0.001). Altogether, 84.6% (44/52) of the respondents did not use protective measures. We described a sudden increase in malaria incidence in a rural, predominantly tea tribe population group with high illiteracy rate and ignorance on protective measures against malaria. More efforts that are concerted needed to educate the community about malaria control practices.

Identification of Major Compounds and α-Amylase and α-Glucosidase Inhibitory Activity of Rhizome of Musa balbisiana Colla: An in-vitro and in-silico Study.

BACKGROUND: α-Amylase and α-glucosidase inhibitors are widely used to suppress postprandial glycemia in the treatment of type-2 diabetes. OBJECTIVES: To evaluate the metallic content, phytocompounds, and α-amylase and α-glucosidase inhibitory activity of Musa balbisiana rhizome using in-vitro and in-silico methods. MATERIALS AND METHODS: Heavy metal content was detected by AAS following standard protocol. Major phytochemicals of the plant were analyzed by GC-MS technique. Enzyme inhibition study was carried out by UV/VIS spectrophotometric methods. The drug-likeness and bio-availability properties of major compounds were carried out using computer-aided tools - SwissADME and ADMElab. Docking and visualization were performed in AutoDock vina and Discovery studio tools. RESULTS: The study found that the fruits of M. balbisiana contain a negligible amount of toxic elements. GC-MS analysis showed five major compounds from the rhizome of M. balbisiana. Invitro enzyme assays revealed strong α-amylase and α-glucosidase inhibitory properties of the plant. All five compounds were predicted to have a drug-likeness property with high cell membrane permeability and bio-availability. The compounds were also predicted to have low to moderate toxicity properties. The Docking study showed strong binding affinities of plant compounds with α-amylase and α-glucosidase. Out of five compounds, C5 showed the best binding affinity with active pockets of α-amylase and α-glucosidase. CONCLUSION: The in-vitro and in-silico study suggests the antihyperglycemic property of the rhizome of Musa balbisiana and a possible candidate for the therapeutic antidiabetic agent(s).

Screening of phytochemicals as potent inhibitor of 3-chymotrypsin and papain-like proteases of SARS-CoV2: an in silico approach to combat COVID-19.

COVID-19 and its causative organism SARS-CoV2 that emerged from Wuhan city, China have paralyzed the world. With no clinically approved drugs, the global health system is struggling to find an effective treatment measure. At this crucial juncture, screening of plant-derived compounds may be an effective strategy to combat COVID-19. The present study investigated the binding affinity of phytocompounds with 3-Chymotrypsin-like (3CLpro) and Papain-like proteases (PLpro) of SARS-CoV2 using in-silico techniques. A total of 32 anti-protease phytocompounds were investigated for the binding affinity to the proteins. Docking was performed in Autodock Vina. Pharmacophore descriptors of best ligands were studied using LigandScout. Molecular dynamics (MD) simulation of apo-protein and ligand-bound complexes was carried out in YASARA software. The druglikeness properties of phytocompounds were studied using ADMETlab. Out of 32 phytochemicals, amentoflavone and gallocatechin gallate showed the best binding affinity to 3CLpro (-9.4 kcal/mol) and PLpro (-8.8 kcal/mol). Phytochemicals such as savinin, theaflavin-3,3-digallate, and kazinol-A also showed strong affinity. MD simulation revealed ligand-induced conformational changes in the protein with decreased surface area and higher stability. The RMSD/F of proteins and ligands showed stability of the protein suggesting the effective binding of the ligand in both the proteins. Both amentoflavone and gallocatechin gallate possess promising druglikeness property. The present study thus suggests that Amentoflavone and Gallocatechin gallate may be potential inhibitors of 3CLpro and PLpro proteins and effective drug candidates for SARS-CoV2. However, the findings of in silico study need to be supported by in vivo studies to establish the exact mode of action.Communicated by Ramaswamy H. Sarma.

In vitro study of the antioxidant, antiproliferative, and anthelmintic properties of some medicinal plants of Kokrajhar district, India.

Alstonia scholaris, Cardiospermum halicacabum, Hydrocotyle sibthorpioides, and Hypericum japonicum are important folk medicinal plants used by tribal communities of Bodoland region of Assam to treat helminth infections. Because of their ethnomedicinal values, the present study was designed to investigate the antioxidant, antiproliferative, and anthelmintic activities of the plants. The antioxidant activity was measured by total antioxidant capacity, total phenolics (TPC), total flavonoid (TFC), FRAP, DPPH, ABTS, and TBARS assay. Antiproliferative and apoptosis-inducing activities of plants were conducted in Dalton's lymphoma (DL) cells. Cells were treated for 24 h with different doses (25-200 mg/mL) of plant extracts. Anthelmintic study was conducted by treating the Paramphistomum sp. at different doses of plant extracts. Phytochemical and antioxidant studies showed rich TPC, TFC, and free radical scavenging activity in H. japonicum and H. sibthorpioides. Both the antiproliferative and anthelmintic bioassays showed a dose-dependent efficacy in all plants. H. japonicum showed the strongest anthelmintic activity (LC50 0.21 mg/mL) followed by H. sibthorpioides (5.36 mg/mL), C. halicacabum (13.40 mg/mL), and A. scholaris (18.40 mg/mL). Evidently, H. sibthorpioides showed the strongest antiproliferative and apoptosis-inducing activities among all the plants. The study observed a positive correlation between the antioxidant properties and antiproliferative and anthelmintic activities of the plants. We, therefore, conclude that the phytocompounds present in the crude extracts along with antioxidant molecules may have combined effects contributing to the antiproliferative and anthelmintic activities of the plants.

Persicaria strigosa (R.Br.) Nakai: a natural anthelmintic?

Persicaria strigosa (R.Br.) Nakai. (Family Polygonaceae) is an important plant of Assam, having several ethnobotanical uses. Tribal communities consume leaf extracts to cure helminth infection. The present study investigated the antioxidant, phytochemicals, and anthelmintic activity of P. strigosa. Total phenolic and flavonoid contents were estimated following standard methods. Antioxidant properties were analyzed by TAC, FRAP, DPPH, ABTS, and TBARS assays. Anthelmintic activity of plant was investigated in Paramphistomum sp. treated with 5 mg/mL, and paralysis and death times were recorded. The most potent solvent extract was performed GC-MS analysis to identify the phytocompounds. Plant extract-treated parasites were further analyzed for biochemical changes. Additionally, molecular docking was performed to study the binding affinities between phytocompounds and enzymes. P. strigosa showed rich phenolics, flavonoids, and antioxidant properties. Ethyl acetate and methanolic extracts showed more powerful antioxidant properties than other extracts. In vitro anthelmintic study found ethyl acetate and diethyl ether the most active extracts. Treated parasites showed a significant decrease in enzyme activity. The highest inhibition was observed in AchE, followed by MDH, LDH, ALP, and ACP. GC-MS study identified 12 probable compounds from the ethyl acetate extract of P. strigosa. Molecular docking showed the strongest binding affinity between the phytocompounds and AchE enzyme (- 7.6 kcal/mol). Overall, compounds C6, C7, and C12 showed better binding affinity compared to other compounds. The in vitro helminth bioassays and biochemical analysis suggest Persicaria strigosa a possible anthelmintic agent. However, isolation and characterization of bioactive compound(s) may promise new drug candidates for helminth infections.

Antioxidant and Antiproliferative Activity of Selected Medicinal Plants of Lower Assam, India: An In Vitro and In Silico Study.

BACKGROUND: The use of medicinal plants for general wellbeing and disease treatment is a common practice among tribal communities of Kokrajhar districts of Assam. However, little works have been done to study the pharmacological aspect of the plants. OBJECTIVES: The present study intends to study the antioxidant and antiproliferative properties of selected medicinal plants used by the tribal communities of the Kokrajhar district of Assam since ancient times. METHODS: Five traditionally important medicinal plants, namely, Cassia fistula, Citrus grandis, Lindernia crustacea, Sacciolepis myosuroides, and Zingiber zerumbet were investigated for antioxidant, antiproliferative (cytotoxic) and apoptosis-inducing potential in the malignant cancer cell line. Phytochemical content, such as phenolic and flavonoid content, were estimated following standard protocol. The methanolic extract of plants was investigated following the phosphomolybdate method (TAC), FRAP, DPPH, ABTS, and TBARS assays. Antiproliferative activities of the plants were carried out by MTT assay in DL and PBMC cells. The apoptotic study was carried out following the acridine orange and ethidium bromide staining method and fluorescent microscopic imaging. Based on the significant (P≤0.05) high apoptotic inducing potential of the plant and to further dissect the molecular mode of action, including downstream biological action, major phytochemicals derived from L. crustacea were investigated for its prospective binding affinity with anti-apoptotic cancer target proteins. RESULTS: Antioxidant studies by FRAP, DPPH, ABTS, and TBARS assay revealed that all five plants contain considerable free radical scavenging activity. C. fistula showed the strongest free radical scavenging activity while the fruit peel extract of C. grandis showed poor activity. The overall antioxidant activities of plants such as TAC, FRAP, DPPH, ABTS, and TBARS may be arranged in decreasing activity as C. fistula > Z. zerumbet > L. crustacea > S. myosuroides > C. grandis. MTT based cell proliferation study showed that all the plants extract significantly (P≤0.05) inhibited cell viability with negligible cytotoxicity (~5-12%) in normal cells. Moreover, L. crustacea showed promising antiproliferative and apoptosis-inducing ability against Dalton's lymphoma. It is worth mentioning that the major bioactive compounds of the most potent plant extract, L. crustacea interacted with anti-apoptotic proteins (cancer target) with higher affinity and the results are compared with reference inhibitors. CONCLUSION: It is worth noting that these plants have the potential to consider for further scientific studies in different cell lines and animal models. Furthermore, isolation and characterization of bioactive compound(s) may promise the discovery of new and valuable drugs candidate to tackle various human diseases.

Phytochemicals, antioxidant, and anthelmintic activity of selected traditional wild edible plants of lower Assam.

OBJECTIVE: Clerodendrum viscosum, Eryngium foetidum, Lippia javanica, and Murraya koenigii are one among the common wild edible plants in Northeast India which are also used as antidiabetic, stomach-ache relieving drugs, etc., The present study was aimed to reveal the phytochemical, antioxidant, and anthelmintic activity of the plants. MATERIALS AND METHODS: The antioxidant capacity of methanolic extract of plants was studied by 1,1-diphenyl-2-picrylhydrazyl (DPPH), ferric reducing antioxidant power, TBARS, and total antioxidant activity (TAA). Total phenolics, flavonoids, Vitamin C, carbohydrate, and protein are also estimated following standard protocols. Anthelmintic activity of the extracts has also been studied in vitro against trematode parasites. RESULTS: The result showed that the methanolic extracts of plants possess a substantial quantity of alkaloids, phenolics, flavonoids, proteins, carbohydrates, and Vitamin C. Phenolics, flavonoids, and Vitamin C contents were found higher in C. viscosum followed by M. koenigii, L. javanica, and E. foetidum. The in vitro antioxidant assays revealed substantial free radical scavenging property in all the plants. TAA increased in the order C. viscosum > M. koenigii > L. javanica > E. foetidum. Similarly, C. viscosum displayed a better antioxidant capacity with IC50 values 29.74 ± 3.63 µg and 148.77 ± 18.38 µg for DPPH and thiobarbituric acid reactive species, respectively. In addition, the plant extracts also showed good anthelmintic activity against Paramphistomum sp. Time taken for paralysis and death were 0:56 ± 0:09 h and 1:35 ± 0:07 h for L. javanica at 50 mg/mL concentration. CONCLUSION: The study therefore suggests the importance of tested plants as a natural source of free radical scavenger and plausible veterinary uses.

Ultrastructural and biochemical alterations in rats exposed to crude extract of Carex baccans and Potentilla fulgens.

The use of plants as a source of medicine is an important component of the health care system in rural India. Carex baccans (Cyperaceae) and Potentilla fulgens (Rosaceae) have been known since ancient times in northeast India for their antitumor, antidiabetic, and antihelmintic properties. The present study was designed to determine the subacute toxicity profile of the root tuber extract of C. baccans and root-peel extract of P. fulgens in Wistar rats. The subacute oral toxicity was conducted using sublethal doses of 40, 50, 100, 150, 200, and 400 mgkg-1 body weights. Surface topographical and ultrastructural observations of liver and intestinal microvilli showed remarkable deformation and disruption, accompanied by quantitative changes in the liver enzymes, i.e., aspartate aminotransferase and alanine aminotransferase in comparison to those of the control group. Apoptotic cell death was observed in the liver cells of rats exposed to both of the plant extracts. A significant increase in splenic lymphocyte count was also observed in rats exposed to the highest concentration of both extracts. The results showed that consumption of the plant extracts at higher doses may cause toxicological effect if treatment continues for a long time.

Potentilla fulgens (Family Rosaceae), a medicinal plant of north-east India: a natural anthelmintic?

The cestode parasite, Raillietina echinobothrida and the trematode, Gastrothylax crumenifer were exposed to the ethanolic root peel extract of Potentilla fulgens, an antiparasitic local medicinal plant of Meghalaya, India, to evaluate the anthelmintic efficacy of the plant. The parasites were incubated in 1, 5, 10, 20, 50 and 100 mg crude alcoholic extract per ml of phosphate buffered saline (PBS) at a temperature of 37 ± 1°C. Paralysis and death were observed at 2.00 ± 0.05 and 2.80 ± 0.06 h for the cestode and 1.21 ± 0.06 and 2.18 ± 0.04 h for the trematode parasites at the highest test concentration of the plant extract. The commercial anthelmintic, Praziquantel (PZQ) showed higher activity at the tested concentration (0.02 mg/ml). To further investigate the efficacy of the plant extract, vital tegumental enzymes of the parasite viz. Acid phosphatase (AcPase), Alkaline phosphatase (AlkPase) and Adenosine triphosphatase (ATPase) were studied. Quantitatively, the total enzyme activity of AcPase, AlkPase and ATPase was found to be reduced significantly by 69.20, 66.43 and 29.63% for R. echinobothrida and 47.96, 51.79 and 42.63% for G. crumenifer, respectively compared to the respective controls; histochemical study also showed reduction in the visible staining of the enzymes. The reference drug, PZQ also showed more or less similar effect like that of the plant extract. The result suggests that phytochemicals of P. fulgens have anthelmintic potential.

Anthelmintic efficacy of ethanolic shoot extract of Alpinia nigra on tegumental enzymes of Fasciolopsis buski, a giant intestinal parasite.

Ethanolic shoot extract of Alpinia nigra, a traditionally used anthelmintic medicinal plant among the Tripuri tribes of north-east India, was tested in vitro to determine its anthelmintic efficacy in gastrointestinal trematode Fasciolopsis buski, using alterations in the activity of vital tegumental enzymes viz. acid phosphatase (AcPase), alkaline phosphatase (AlkPase) and adenosine triphosphatase (ATPase). Live adult F. buski treated in vitro with different concentrations of the plant extract showed significant decline in the visible stain histochemically and enzyme activities. Quantitatively, the total enzyme activity of AcPase, AlkPase and ATPase was found to be reduced by 45, 41 and 43%, respectively compared to the control. The reference drug, praziquantel also showed more or less similar effect like that of the plant extract. The results suggest that the tegumental enzymes of the parasite may be an important target of action for active component(s) of A. nigra, which appears to act transtegumentally.