Wound healing mechanisms of Couroupita guianensis fruit pulp: An ethnomedicine used by traditional healers in India.

In connection to search for safe and alternative plant-based drugs, the wound healing mechanisms of an Indian ethnomedicine Couroupita guianensis fruit pulp was analyzed in this project work. Gas chromatography coupled with mass spectrometer (GC-MS) analysis revealed the existence of phytochemicals such as 2-furoic acid, 2,4-heptadienal, pyrazole and 8-hydroxyquinoline in the methanol extract. Methanol extract of C. guianensis exhibited remarkable radical scavenging activity against 2, 2-diphenyl-1-picrylhydrazyl (DPPH) (89.88%), superoxide (91.51%), hydrogen peroxide (24.25%) and hydroxyl radicals (73.62%). Further, it showed remarkable anti-inflammatory (24.09-62.16%) and anti-bacterial activity (zone of inhibition, ZOI: 13.00 mm, minimum inhibitory concentration, MIC: 6.25 mg/mL and minimum bactericidal concentration, MBC: 12.51 mg/mL) and also controlled the growth rate of methicillin resistant Staphylococcus aureus (MRSA) within 30 min of treatment. The angiogenic potential of C. guianensis was proved in chick chorioallantoic membrane (CAM) model and it does not exhibit any toxicity in peripheral blood monocyte cells (PBMC) model.

Vitexin isolated from Prosopis cineraria leaves induce apoptosis in K-562 leukemia cells via inhibition of the BCR-ABL-Ras-Raf pathway.

OBJECTIVES: Leukemia is one of the severe cancer types all around the globe. Even though some chemotherapeutic drugs are available for treating leukemia, they have various side effects. As an alternative approach, herbal drugs are focused on current research to overcome leukemia. The present work was conducted to investigate the antileukemic mechanism of active phytochemical vitexin, which was isolated from ethno-medicine (Prosopis cineraria leaf) used by traditional healers of West Bengal, India. METHODS: Antiproliferative mechanisms of selected phyto-compound against K-562 cells were evaluated using cellular uptake, morphological changes, DNA fragmentation, mitochondrial membrane potential and signaling pathways analysis. KEY FINDINGS: Vitexin exhibited cytotoxicity by reducing mitochondrial membrane potential (32.40%) and causing DNA fragmentation (84.15%). The western blotting study indicated inhibition of cell survival proteins (BCR, ABL, H-RAS, N-RAS, K-RAS and RAF) and expression of apoptotic proteins (p38, BAX and caspase-9) in leukemia cells upon treatment with vitexin. CONCLUSIONS: Based on the results, presently investigated phyto-compound vitexin could be considered for developing safe and natural drugs to treat leukemia after conducting suitable preclinical and clinical trials.

Antioxidant and cytoprotective properties of loganic acid isolated from seeds of Strychnos potatorum L. against heavy metal induced toxicity in PBMC model.

Present work carried out with the objectives to isolate active component of S. potatorum and also to evaluate its free radical scavenging activity and preventing capacity against heavy metal toxicity. Solvents of different polarity were used to prepare crude extracts of S. potatorum seeds and screened for antioxidant activity. Among the crude extracts, methanolic extract was found to exhibit higher antioxidant activity (81.22%) which was fractionated by liquid-liquid partitioning method. Among the different fractions (LF1-LF4), LF-2 showed higher antioxidant activity (98.24%) as compared to other three liquid fractions and hence LF-2 was further purified by column chromatography. Among nine column fractions (CF1-CF9), fraction CF-7 was found to have higher antioxidant activity (92.14%), which was further analyzed using LC-MS and NMR and identified as loganic acid. In vitro radical scavenging assays showed remarkable antioxidant activity of loganic acid in terms of DPPH scavenging (IC50 149 µg/ml), superoxide radical scavenging (IC50 632.43 µg/ml) and hydroxyl radical scavenging (IC50 29.78 µg/ml). Loganic acid exhibited 81% prevention of heavy metal toxicity through the mechanism of inhibiting ROS generation (2046 AU vs. 5264 AU in control) and lipid peroxidation (95.01%). Thus, the active compound (loganic acid) isolated from S. potatorum has strong free radical scavenging activity and remarkable cyto-protective effect against heavy metal mediated toxicity.

Apoptotic mechanisms of myricitrin isolated from Madhuca longifolia leaves in HL-60 leukemia cells.

Myricitrin, a naturally occurring flavonoid in Madhuca longifolia, possesses several medicinal properties. Even though our earlier work revealed its role against the proliferation of acute myelogenous leukemia cells (HL-60), its molecular mechanisms have not yet been revealed. The current study aims to explore the molecular mechanisms of myricitrin (isolated from an ethnomedicinal drug Madhuca longifolia) to induce apoptosis in HL-60 cells. Treatment with IC-50 dose of myricitrin (353 µM) caused cellular shrinkage and cell wall damage in HL-60 cells compared to untreated control cells. Myricitrin treatment reduced the mitochondrial membrane potential (22.95%), increased DNA fragmentation (90.4%), inhibited the cell survival proteins (RAS, B-RAF, & BCL-2) and also induced pro-apoptotic proteins (p38, pro-caspase-3, pro-caspase-9 and caspase-3) in the HL-60 cells. The present study provides scientific evidence for the apoptosis caused by myricitrin in HL-60 leukemia cells. Hence, the phytochemical myricitrin could be considered as a potential candidate to develop an anticancer drug after checking its efficacy through suitable pre-clinical and clinical studies.

Oxidative stress mediated cytotoxicity in leukemia cells induced by active phyto-constituents isolated from traditional herbal drugs of West Bengal.

ETHNOPHARMACOLOGICAL RELEVANCE: In search of safe and effective therapeutic agents as alternative to synthetic chemotherapeutics for the treatment of leukemia, the herbal drugs (Leaf of Madhuca longifolia, leaf of Prosopis cineraria and bark of Flacourtia indica) with long traditional use in West Bengal have received our attention. AIM OF THE STUDY: Present work was conducted to isolate and identify the active compounds of the selected herbal drugs using bio-assay guided fractionation and also to investigate their anticancer mechanism in leukemia cell lines. MATERIALS AND METHODS: Bio-assay guided fractionation was used for the isolation of active constituents such as myricitrin, vitexin and vanillin from the aqueous extracts of M. longifolia, P. cineraria and F. indica, respectively using liquid partitioning and column chromatography and the compounds were characterized by HPLC, MS and NMR. Dose and time-dependent cytotoxicity of isolated compounds were studied against leukemia cells and their anticancer mechanism such as cell wall damage, nuclear damage, ROS and NO generation, SOD level, LDH release and lipid peroxidation were investigated. RESULTS: Aqueous extract of M. longifolia, P. cineraria and F. indica exhibited maximum anti-proliferative activity against HL-60 (Acute myeloid leukemia, AML, 72.06%), K-562 (Chronic myeloid leukemia, CML, 42.14%) and Jurkat (Acute lymphoblastic leukemia, ALL, 51.71%) cells. Myricitrin, vitexin and vanillin exhibited dose-dependent (IC-50 values 164.4, 147 & 29.22 µg/ml) and time-dependent activity with maximum cytotoxicity at 48 h. All these three compounds caused apoptosis in leukemia cells by inducing free radicals such as ROS (1.33-2.65 Arbitrary units) and NO (11.17-18.53 µM), cell membrane damage and nuclear condensation which were evidenced by increased release of LDH (1326-1439 U/L), improved lipid peroxidation (10.19-14.41 nM/mg protein) and reduced SOD level (6.2-9.21 U/mg protein) in leukemia cells. CONCLUSIONS: Based on anti-proliferative activity, the isolated phyto-compounds myrcitrin, vitexin and vanillin from M. longifolia, P. cineraria and F. indica could be developed as natural drugs for treating AML, CML and ALL leukemia types, respectively.

Investigation of phytochemical constituents of anti-leukemic herbal drugs used by the traditional healers of Purulia, Birbhum and Bankura districts of West Bengal.

In the present work, 16 different plant drugs used by traditional healers from West Bengal were screened through in vitro cell line model. Herbal drugs used by traditional tribal healers in Purulia, Birbhum and Bankura districts of West Bengal were collected and screening against acute myeloid leukemia (AML) cell line (HL-60). Among 16 plant extracts, bark of Flacourtia indica (66.67%), leaf of Madhuca longifolia (69.17%), and leaf of Prosopis cineraria (68.08%) showed better cytotoxicity results than other herbals. Further, time-dependent study showed maximum cytotoxicity of the selected herbal extracts between 36 and 48 hours of treatment in both acute and chronic myeloid leukemia (CML) cell lines (HL-60 and K562). The LC-MS/MS analysis of the selected drugs revealed the presence of picrotoxinin and 10-deacetylbaccatin from F. indica, isoorientin and hirsutrin from M. longifolia, vitexin and rhoifolin in P. cineraria.

Agro food by-products and essential oil constituents curtail virulence and biofilm of Vibrio harveyi.

Vibrio harveyi causes severe loss to the aquaculture industry due to its virulence, which is mediated by Quorum sensing (QS) and biofilm formation. In the current study, we have explored the anti-virulent properties and biofilm disruption ability of luteolin (extracted from coconut shell) and linalool against this important aquaculture pathogen. HPLC analysis of the methanolic extract of coconut shells revealed a single major peak which matched to the standard luteolin which was further elucidated by NMR studies. Further, luteolin and linalool were screened for their ability to inhibit biofilms and various quorum sensing mediated virulence factors of V. harveyi. The Minimum Inhibitory Concentration (MIC) of the two compounds was determined and the sub-inhibitory concentrations of the compounds were able to inhibit biofilm formation. Both the compounds disrupted about 60-70% mature biofilms, which was also visually observed by light microscopy. Both linalool and luteolin exhibited a significant reduction in the production of EPS and alginate in the biofilms matrix of V. harveyi which was confirmed by Scanning Electron Microscopy (SEM). Both compounds inhibited the swarming and swimming motility, the crucial quorum sensing (QS) mediated virulence of V. harveyi. The present study shows the presence of valuable polyphenolic compound like luteolin in coconut shells that are discarded as a waste. From the present study we envisage that luteolin and linalool can serve as potent anti-virulent agents to combat QS mediated infections against aquaculture pathogens.

Green synthesis of silver nanoparticles using Nardostachys jatamansi and evaluation of its anti-biofilm effect against classical colonizers.

In this communication, we present the green synthesis of silver nanoparticles (AgNPs) using medicinally important Nardostachys jatamansi rhizome extract in the presence of sunlight. UV-vis spectroscopy, Fourier Transform Infrared spectroscopy (FTIR), Transmission electron microscope (TEM) and Energy dispersive X-ray analysis (EDX) were employed to characterize the synthesized AgNPs. UV-visible spectroscopic studies confirmed the presence of biosynthesized AgNPs. Transmission Electron Microscopic studies revealed the structure of spherical AgNPs in the diameter range of 10-15 nm. Energy dispersive X-ray analysis and elemental mapping clearly confirmed the presence of silver in AgNPs samples. Interestingly, biomolecules functionalised AgNPs exhibited a remarkable antioxidant, anti-inflammatory, and anti-biofilm activities and hence biosynthesized AgNPs from N. jatamansi can be used as a promising biomaterial for biomedical applications.

Potential anti-proliferative activity of AgNPs synthesized using M. longifolia in 4T1 cell line through ROS generation and cell membrane damage.

To overcome the problem of breast cancer, silver nanoparticles (AgNPs) synthesized using Indian medicinal plant Madhuca longifolia could be explored as an alternative anticancer medicine. Synthesized AgNPs were studied their characteristics and their anti-proliferative property was investigated in breast cancer cell line (4T1). Based on zeta sizer analysis, the size of the AgNPs was 103.5 nm and potential -9.57 eV. Fe-SEM results showed particle size of 69.4-99.4 nm while TEM images indicated the particle size of 18-24 nm. In dose-dependent study, AgNPs showed 93% of anti-proliferative activity at 50 µg/ml whereas the methanolic extract of M. longifolia showed 80% activity only at 10-fold increased concentration (500 µg/ml). AgNPs exhibited higher level of cytotoxicity in breast cancer cell line than extract through cell wall degradation and ROS generation. Such effective AgNPs could be investigated further through in vivo models with a view to develop anticancer drug.

Studies on physicochemical and nutritional properties of aerial parts of Cassia occidentalis L.

In the present, work chemical composition and nutritional value of aerial parts of Cassia occidentalis L. was studied. The aerial parts of C. occidentalis possess favorable physicochemical properties with good nutritional value, such as high energy value, crude fibers, and vitamin levels. The X-ray fluorescence spectrophotometry data revealed that the sample is rich in minerals, especially in Fe, Ca, K, and Mn. Further, minerals such as Mg, Zn, Cu, Na, P, and S are present in good amount and depicted the nutritional value of the selected material. The plant sample is rich in phytochemicals such as flavonoids, alkaloids, lignin, tannins, and phenols. The presence of phytochemical constituents was confirmed by gas chromatography-mass spectrometry profile and high-performance thin layer chromatography fingerprinting techniques. The findings stimulate the on-farm cultivation of C. occidentalis on a large scale to relieve the iron deficiency in local community, and it can be used as a dietary supplement to treat anemia.

Development, acceptability, and nutritional characteristics of a low-cost, shelf-stable supplementary food product for vulnerable groups in Kenya.

BACKGROUND: Food-based approaches have been advocated as the best strategies to curb hunger and malnutrition in developing countries. The use of low-cost, locally available, nutritious foods in the development of supplementary foods has been recommended. OBJECTIVE: To develop low-cost food supplements using different traditionally processed local foods, consisting of cereals, legumes, nuts, fish, and vegetables, to meet the nutrient requirements for vulnerable groups in Kenya. METHODS: Four food supplements were developed and evaluated by taste panel procedures. The product containing amaranth grain, pigeon pea, sweet potato, groundnuts, and brown sugar was found to be the most acceptable supplement. Evaluation of nutritional composition, shelf-life, and cost analysis of the acceptable supplement was carried out to assess if it could satisfactorily provide more than 50% of the Recommended Dietary Allowances (RDAs) of the basic nutrients for vulnerable groups. RESULTS: The acceptable supplement contained 453.2 kcal energy, 12.7 g crude protein, 54.3 g soluble carbohydrates, 20.8 g crude fat, and 10.1 g crude fiber per 110 g. The micronutrient contents were 93.0 mg calcium, 172.4 mg magnesium, 2.7 mg zinc, 5.7 mg iron, 0.8 mg vitamin B1, 0.2 mg vitamin B2, 7.9 mg niacin, 100 microg folic acid, and 140 microg retinol equivalent per 110 g. The supplement also contained 21% total essential amino acid in addition to appreciable levels of palmitic, stearic, oleic, linoleic, and alpha-linolenic fatty acids. The shelf-life study showed that it could be stored in different packaging materials (polythene bags, gunny bags, and kraft paper) at 26°C without deleterious effects on its chemical composition for up to 4 months. Cost analysis of the supplement indicated that the product could be competitively sold at US$0.812/kg (KES 65.50/kg). CONCLUSIONS: Locally available indigenous foods can be used in the formulation of acceptable, low-cost, shelf-stable, nutritious supplementary foods for vulnerable groups.

Bioactive compounds extracted from Indian wild legume seeds: antioxidant and type II diabetes-related enzyme inhibition properties.

Seven different wild legume seeds (Acacia leucophloea, Bauhinia variegata, Canavalia gladiata, Entada scandens, Mucuna pruriens, Sesbania bispinosa and Tamarindus indica) from various parts of India were analyzed for total free phenolics, l-Dopa (l-3,4 dihydroxyphenylalanine), phytic acid and their antioxidant capacity (ferric-reducing antioxidant power [FRAP] and 2,2-diphenyl-1-picrylhydrazyl [DPPH] assay) and type II diabetes-related enzyme inhibition activitiy (α-amylase). S. bispinosa had the highest content in both total free phenolics and l-Dopa, and relatively low phytic acid when compared with other seeds. Phytic acid content, being highest in E. scandens, M. pruriens and T. indica, was highly predictive for FRAP (r = 0.47, p < 0.05) and DPPH (r = 0.66, p < 0.001) assays. The phenolic extract from T. indica and l-Dopa extract from E. scandens showed significantly higher FRAP values among others. All seed extracts demonstrated a remarkable reducing power (7-145 mM FeSO4 per mg extract), DPPH radical scavenging activity (16-95%) and α-amylase enzyme inhibition activity (28-40%).

Antioxidant and type 2 diabetes related functional properties of phytic acid extract from Kenyan local food ingredients: effects of traditional processing methods.

Emerging scientific evidences reveal that phytic acid has several positive effects on human health. The antioxidant and type 2 diabetes related enzyme inhibition properties of phytic acid extract prepared from raw and traditionally processed local grains and vegetables collected from Kenya were evaluated. Phytic acid content of raw grains and vegetables ranged between 2.81-3.01 and 0.29-3.23 g/100 g DM, respectively. The phytic acid extract from raw samples revealed 59%-89% of DPPH radical scavenging capacity, 27-3,526 mmol Fe(II)/g extract of reducing power, 20%-72% of α-amylase inhibition activity and 8%-91% of α-glucosidase inhibition activity. Cooking and roasting improved the antioxidant and health relevant functionality of phytic acid extracts obtained from Kenyan local vegetables and grains, respectively.

Antioxidant potential and health relevant functionality of traditionally processed Cassia hirsuta L. seeds: an Indian underutilized food legume.

The methanolic extract of Cassia hirsuta L. seed materials, an underutilized food legume collected from India, was analyzed for antioxidant activity and health relevant functionality. The methanolic extract of raw seeds contained a total free phenolic content of 15.82 ± 1.69 g catechin equivalent/100 g extract DM. Encouraging levels of ferric reducing/antioxidant power (FRAP, 1,446 mmol Fe[II]/mg extract), inhibition of ß-carotene degradation (48.81%) and scavenging activity against DPPH (64.40%) and superoxide (43.78%) radicals were exhibited by the raw samples. Further, 83.11% of α-amylase and 62.79% of α-glucosidase enzyme inhibition characteristics under in vitro starch digestion bioassay were also recorded. Sprouting + oil-frying caused an apparent increase on the total free phenolic content and a significant improvement in the antioxidant and free radical scavenging capacity of methanolic extract of C. hirsuta seeds, while soaking + cooking as well as open-pan roasting treatments showed diminishing effects. The analysis of the phenolic profile revealed the presence of gallic acid, p-coumaric acid and (+)-catechin in the methanolic extract of these seeds.

Flavonoid content in ethanolic extracts of selected raw and traditionally processed indigenous foods consumed by vulnerable groups of Kenya: antioxidant and type II diabetes-related functional properties.

The present study evaluated the flavonoid content, antioxidant as well as type II diabetes-related enzyme inhibition activities of ethanolic extract of certain raw and traditionally processed indigenous food ingredients including cereals, legumes, oil seeds, tubers, vegetables and leafy vegetables, which are commonly consumed by vulnerable groups in Kenya. The vegetables exhibited higher flavonoid content (50-703 mg/100 g) when compared with the grains (47-343 mg/100 g). The ethanolic extract of presently studied food ingredients revealed 33-93% DPPH radical scavenging capacity, 486-6,389 mmol Fe(II)/g reducing power, 19-43% α-amylase inhibition activity and 14-68% α-glucosidase inhibition activity. Among the different food-stuffs, the drumstick and amaranth leaves exhibited significantly higher flavonoid content with excellent functional properties. Roasting of grains and cooking of vegetables were found to be suitable processing methods in preserving the functional properties. Hence, such viable processing techniques for respective food samples will be considered in the formulation of functional supplementary foods for vulnerable groups in Kenya.

Antioxidant and antidiabetic properties of condensed tannins in acetonic extract of selected raw and processed indigenous food ingredients from Kenya.

Recently, tannins have received considerable attention as health-promoting component in various plant foods and several studies have reported on its nutraceutical properties. However, no study has established the role of condensed tannins in indigenous foods of Kenya. Therefore, this study was designed to evaluate the antioxidant activity (DPPH and FRAP) and antidiabetic effects (α-amylase and α-glucosidase inhibition activities) of condensed tannins in some selected raw and traditionally processed indigenous cereals, legumes, oil seeds, and vegetables. The condensed tannin content of the grains and vegetables ranged between 2.55 and 4.35 g/100 g DM and 1.53 and 5.73 g/100 g DM, respectively. The scavenging effect of acetonic extract on DPPH radical ranged from 77% to 90% while the reducing power was found to be 31 to 574 mmol Fe(II)/g DM in all the investigated food ingredients. The condensed tannin extracts of the analyzed samples showed promising antidiabetic effects with potential α-amylase and α-glucosidase inhibition activities of 23% to 44% and 58% to 88%, respectively. Condensed tannins extracted from the amaranth grain, finger millet, field bean, sunflower seeds, drumstick, and amaranth leaves exerted significantly higher antioxidant and antidiabetic activities than other food ingredients. Among the traditional processing methods, roasting of grains and cooking of vegetables were found to be more suitable mild treatments for preserving the tannin compound and its functional properties as opposed to soaking + cooking and blanching treatments. The identified elite sources of optimally processed indigenous food ingredients with promising results could be used as health-promoting ingredients through formulation of therapeutic diets.