Bioassay guided fractionation and identification of active anti-inflammatory constituent from Delonix elata flowers using RAW 264.7 cells.

CONTEXT: Delonix elata (L.) Gamble (Fabaceae) has been used in the Indian traditional medicine system to treat rheumatism and inflammation. AIM: To assess the anti-inflammatory effect of Delonix elata flowers and to isolate the active principle. MATERIALS AND METHODS: The prompt anti-inflammatory constituent was isolated from Delonix elata flower extracts using bioassay guided fractionation in liposaccharide (LPS) stimulated RAW 264.7 macrophage cell line. The anti-inflammatory activity of extracts/fractions/sub-fractions/compounds (10, 25, and 50 µg/ml) was evaluated by estimating the levels of nitric oxide (NO), TNF-α, and IL-1ß after 24 h of LPS induction (1 µg/ml). The isolated active compound was subjected to NMR, IR, and UV analyses for structure determination. RESULTS: In an attempt to search for anti-inflammatory constituents, the active pure principle was isolated and crystallized as a white compound from Delonix elata flowers methanol extract. This active compound (50 µg/ml) decreased the release of inflammatory mediators levels such as NO (0.263 ± 0.03 µM), TNFα (160.20 ± 17.57 pg/ml), and IL-1ß (285.79 ± 15.16 pg/ml) significantly (p < 0.05); when compared to the levels of NO (0.774 ± 0.08 µM), TNFα (501.71 ± 25.14 pg/ml), and IL-1ß (712.68 ± 52.25 pg/ml) from LPS-stimulated macrophage cells. The active compound was confirmed as hesperidin with NMR, IR, and UV spectroscopy data. This is the first report of this compound from Delonix elata flowers. CONCLUSION: The findings of the study support the traditional use of Delonix elata flowers to treat inflammation.

Myroides pelagicus from the gut of Drosophila melanogaster attenuates inflammation on dextran sodium sulfate-induced colitis.

BACKGROUND AND AIM: Probiotics are live microorganisms that confer a health benefit on the host when administered in adequate amounts. In the present study, the putative probiotic strain was identified from the gut of Drosophila melanogaster and assessed for its protective effect in inflammatory bowel disease. METHODS: Active probiotics were screened from the Drosophila melanogaster gut by the selection criteria of gastric juice tolerance, hydrophobic property, antimicrobial potential, adhesion, and invasion properties. The active probiotics were identified by 16s rDNA sequencing and the effect of these active probiotics was evaluated in a Dextran sulphate sodium (DSS)-induced mice model by estimating inflammatory markers and histopathological changes. RESULTS: Nine Gram-positive and bile salt tolerant bacterial isolates were obtained from the gut samples. The isolates PTH 2, PTH 4, and PTH 7 clearly showed significant activity in antimicrobial potential, hydrophobic (>74%) property, and intestinal juice tolerance. Among these, PTH 7 was selected for further studies due to its significant low-invasion ability and it proved capable of reducing the secretion of proinflammatory cytokines. The 16s rDNA studies revealed that PTH 7 was Myroides pelagicus. Administration of M. pelagicus to the DSS-induced colitic animals significantly suppressed myeloperoxidase, ALP, and malondialdehyde levels, and also lowered levels of proinflammatory cytokine expression. Further, the recovery from the disease by the probiotic treatment was supported by histopathological and macroscopic observation. The treated animals did not show any adverse signs in their physiology or behavior. CONCLUSION: Myroides pelagicus successfully prohibited inflammatory markers and acted as a potent probiotic. Future studies with this stain might prove its efficacy as a drug for the management of colitis.

A study on food-medicine continuum among the non-institutionally trained siddha practitioners of Tiruvallur district, Tamil Nadu, India.

BACKGROUND: Medicinal properties of the food species are one of the poorly documented and important areas of ethnopharmacology. The present survey quantitatively documented the medicinal foods prescribed by the non-institutionally trained siddha practitioners of Tiruvallur district of Tamil Nadu. METHODS: Field work was carried out between December 2014 and April 2017 using a questionnaire. The illnesses mentioned by the informants were grouped as illness categories on the basis of emic perceptions. Sufficiency of sampling of this survey was assessed by plotting the cumulative number of UR and Shannon-Wiener's index. The indices such as informant consensus factor (FIC), Index of Agreement on Remedies (IAR), and Cultural Food Significance Index (CFSI) were calculated. RESULTS: This study documented 165 medicinal foods used by 82 non-institutionally trained siddha practitioners of Tiruvallur district, and 73.93% of these preparations were plant based. Among the animal taxa, 82.05% were represented by fish taxa. The illness category gastrointestinal ailments is the majorly cited illness category treated with plant-based formulations. The illness categories viz., gastrointestinal ailments, hemorrhoids, and neural ailments had high consensus under the group of plant-based medicinal foods. In animal-based medicinal foods, kapha ailments had gained 23.07% of UR. The illness categories such as bone fractures, male reproductive ailments, blood ailments, and anabolic had high FIC values. CONCLUSIONS: Deeper studies on different dietary cultures of India may help to derive better interpretations on food-medicine continuum. This study identified some important claims such as the use of citron, pomegranate and Solanum americanum (gastrointestinal ailments), Abutilon indicum, onions and elephant foot yam (hemorrhoids), Boerhavia diffusa (urinary ailments), Moringa oleifera (anemia), Aloe vera (gynecological ailments), Eclipta prostrata (liver ailments), ivy gourd (diabetes), citron (hypertension), Centella asiatica (psychological ailments), spade nose shark (lactogogue), reticulate whipray (wheezing and bronchitis), Katelysia opima (impotence), Indian squid (anemia), and Indian oil sardine (anabolic). More studies on these claims will help identify novel functional foods to add to the field of medical nutrition therapy, with traditional brand identity. Robust studies on the documentation of the traditional knowledge on marine resources will yield a good database for various stakeholders and policy makers.

Swertiamarin, a natural steroid, prevent bone erosion by modulating RANKL/RANK/OPG signaling.

Bone erosion is a central feature of rheumatoid arthritis (RA) that is characterized by the infiltration of the synovial lining by osteoclasts and lymphocytes. In the present study, swertiamarin a major secoiridoid glycoside was evaluated for anti-osteoclastogenic property to prevent bone erosion in Freund's complete adjuvant (FCA) induced in-vivo model, in-vitro osteoblast and osteoclasts as well as in co-culture system and in-silico molecular docking analysis. The swertiamarin treatment decreased the expression of TRAP, RANKL, and RANK levels and increased the levels of OPG levels significantly in both in vitro and in vivo models. In in vitro, the compound treatment significantly increased the cell proliferation and ALP levels in osteoblast cells; the high proliferation (153.8600±5.23%) and ALP release (165.6033±4.13%) were observed at 50µg/ml concentration of swertiamarin treatment. At the same time the treatment decreased the TRAP positive cells in osteoclast cells; the high reductions of TRAP positive cells (39.32±3.19%) were observed at 50µg/ml of swertiamarin treatment. The treatment modulated the levels of pro-inflammatory cytokines, MMPs and NF-κB levels in osteoblast and osteoclast co-culture system. In in silico analysis swertiamarin had affinity towards the proteins RANK, RANKL and OPG residues with low binding energy -4.5, -3.92 and -5.77kcal/mol respectively. Thus, the results of this study revealed the anti-osteoclastogenic activity of swertiamarin on the prevention of bone destruction.

Facultative alkalophilic bacteria from mangrove soil with varying buffering capacity and H+ conductance.

Facultative alkalophilic bacteria Planococcus sp. (EMGA-26), Bacillus sp. (EMGA-29) and Corynebacterium spp. (EMGA-33 and 130) were isolated from mangrove soil samples. Neutrophiles were predominant than alkalophiles. Buffering capacity and membrane H+ conductance were investigated for the strains grown in PPYG medium at pH 10.5 using acid pulse technique. Bacillus sp. showed higher buffering capacity than Planococcus sp. and Corynebacterium spp. Buffering capacity was two-fold higher in Corynebacterium sp. EMGA-33 than in EMGA-130. The membrane H+ conductance was high in Bacillus sp. and was directly proportional to the buffering capacity values. The Bacillus sp. (EMGA-29) had higher cell membrane adaptability in high pH environment than the Planococcus sp. and Corynebacterium spp.

In vivo and in vitro immunomodulatory potential of swertiamarin isolated from Enicostema axillare (Lam.) A. Raynal that acts as an anti-inflammatory agent.

Swertiamarin is a secoiridoid glycoside found in Enicostema axillare (Lam) A. Raynal, a medicinal plant used as a depurative in the Indian system of traditional medicine. The present study evaluated the immunomodulatory activity of isolated swertiamarin. In vivo immunomodulatory activity of swertiamarin (2, 5, and 10 mg/kg b.w.) was evaluated in a model of sheep red blood cells (SRBC) by assessing its effect on organ weight, hemagglutinating antibody titer (HA), plaque-forming cells (PFC), quantitative hemolysis of SRBC, and delayed type hypersensitivity (DTH). In vitro immunomodulatory potential was studied on isolated splenocytes, neutrophils, and peritoneal macrophages. In silico immunomodulatory effects were evaluated by docking of swertiamarin on proinflammatory cytokines to confirm its potential. In in vivo studies, the animals treated with swertiamarin showed a significant (P ≤ 0.05) increase in antibody titer, plaque-forming cells, and also in weight of the thymus and spleen. A decreased response to DTH reaction was recorded with the treatment of swertiamarin. In in vitro studies, treatment with swertiamarin modulated the messenger RNA (mRNA) and protein expression of IFN-γ, IL-10, and IL-4 significantly (P ≤ 0.05) and also favored Th2-mediated response on concanavalin A (Con A)-induced splenocytes. The compound inhibited the release of free radicals significantly (P ≤ 0.05) in phytohemagglutinin (PHA)-induced neutrophils and also ameliorated the mRNA and protein expression of proinflammatory cytokines (TNF-α, IL-1ß, and IL-6) in lipopolysaccharide (LPS)-induced macrophages. In in silico, the best docked pose of swertiamarin with the target proteins (TNF-α, IL-1ß, and IL-6) was confirmed that swertiamarin acted as an anti-inflammatory mediator.

Documentation and quantitative analysis of local ethnozoological knowledge among traditional healers of Theni district, Tamil Nadu, India.

ETHNOPHARMACOLOGICAL RELEVANCE: This study investigated the use of animals among the traditional healers in Theni district of Tamil Nadu, India. The data regarding the medicinal animals/animal products were documented and their usages were analyzed quantitatively. MATERIALS AND METHODS: Based on free list interviews with the traditional healers, we documented the medicinal usage of animals/animal products and calculated the indices such as informant consensus factor (Fic) to determine the consensus over the species for an illness category, as well as the Index Agreement on Remedies (IAR) to determine the extent of potential utilization of each species. RESULTS: In this study, 69 medicinal animals/animal products were documented with the help of standardized questionnaires among the local healers. The results were tabulated and Fic value for each illness category was calculated. Three illness categories viz., jaundice (milk of Capra aegagrus hircus), orthopedics (egg white and meat of Gallus gallus domesticus) and pediatrics (milk of Equus africanus asinus) had got high Fic values. Fifteen illness categories had moderate Fic values. Highly cited animals in these illness categories were: Rusa unicolor (antiemetic), Reticulitermes spp. (diabetes), flesh of Varanus benghalensis (oral ailments), milk (eye ailments, fever) and urine (antidote) of Homo sepians, meat of Trachypithecus johnii (respiratory ailments), various parts of C. aegagrus hircus (blood ailments, coolants, diarrhea, pulmonary and urinary ailments), flesh of Chamaeleon zeyalnica (neural ailments), meat of Passer domesticus (aphrodisiac), curd and dung of Bos primigenius taurus (dermatological ailments), meat of G. domesticus (musculo-skeletal disorders, analgesic), meat of Lissemys punctata (hemorrhoids), and Pherthima posthuma (psychological ailments). Six illness categories had low Fic values. CONCLUSION: This study indicated that the animals are still being used by the local healers of Theni district, to treat various illnesses. Cross-disciplinary approaches to explore the full potential of animal-derived medicines will help to improve the health of local people.

Ameliorative effect of Drynaria quercifolia (L.) J. Sm., an ethnomedicinal plant, in arthritic animals.

Drynaria quercifolia (L.) J. Sm., is an ethnomedicinal plant used widely in Tamil Nadu to treat arthritis. The present study was aimed to evaluate the traditional claim of D. quercifolia rhizome water extract in adjuvant induced arthritic animals. Anti-arthritic effect was studied by assessing the levels of lysosomal enzymes, protein bound carbohydrates, urinary degradative collagen and serum cytokines on control and adjuvant induced arthritis. The paw swelling and body weight were also analyzed. The levels of ROS and lysosomal enzymes in neutrophils of control and adjuvant induced animals were also estimated. D. quercifolia rhizome water extract at doses of 100 and 200mg/kg reduced the paw thickness and elevated the mean body weight of arthritic rats. The treatment with extract showed a significant reduction in the levels of plasma and liver lysosomal enzymes as well as protein bound carbohydrates and urinary degradative collagen levels. The treatment reduced the levels of ROS and lysosomal enzymes in neutrophils significantly. The significant reduction in the levels of serum pro-inflammatory cytokines (TNF-α and IL-1ß) and the increment in the levels of anti-inflammatory cytokine (IL-10) were also observed by the treatment. The present study supports the traditional claim of using D. quercifolia to treat rheumatism.

Immunomodulatory potential of Enicostema axillare (Lam.) A. Raynal, a traditional medicinal plant.

ETHNOPHARMACOLOGICAL RELEVANCE: Enicostema axillare (Lam.) A. Raynal., (Gentianaceae) has been used in traditional Indian system of medicine as depurative and for the treatment of psoriasis, intermittent fever and cancer. Ethnobotanical survey conducted in Theni District indicated a high consensus for this plant as blood purifier and to treat dermatopathy and venereal infections. The present study evaluated the immunomodulatory activity of the methanol extract of Enicostema axillare. MATERIALS AND METHODS: In vivo immunomodulatory activity of Enicostema axillare methanol extract (100 and 200mg/kg b.w) was evaluated by assessing its effect on the total and differential leukocyte count, organ weight, hemaggultinating antibody titer, plaque forming cells, quantitative hemolysis of SRBC and delayed type hypersensitivity. Sheep red blood cells (SRBC, 5×10(8)cells/0.1mL) were used to immunize the animals. In vitro immunomodulatory potential of the extract was studied using peritoneal macrophages by evaluating its effects on NBT reduction, NO production and cytokine release. RESULTS: The animals treated with Enicostema axillare methanol extract showed a significant (P≤0.05) increase in weight of the thymus and spleen. The total leukocyte and lymphocyte count was increased significantly (P<0.005) by the treatment. There was no significant alteration in neutrophil count. A dose dependent increase in antibody titer value was observed. A decreased response to DTH reaction induced by SRBC was recorded. A potential phagocytic response was seen on treatment with the extracts at 10 and 25µg/mL. The extract inhibited the release of pro-inflammatory cytokines and production of NO significantly in a dose dependent manner. CONCLUSION: These findings suggested that the methanol extract of Enicostema axillare acted on both humoral and cell mediated immune functions and decreased the release of pro-inflammatory cytokines in the peritoneal macrophages.

Antifeedant and larvicidal activities of Rhein isolated from the flowers of Cassia fistula L.

Antifeedant and larvicidal activities of rhein (1,8-dihydroxyanthraquinone-3-carboxylic acid) isolated from the ethyl acetate extract of Cassia fistula flower were studied against lepidopteron pests Spodoptera litura and Helicoverpa armigera. Significant antifeedant activity was observed against H. armigera (76.13%) at 1000 ppm concentration. Rhein exhibited larvicidal activity against H. armigera (67.5), S. litura (36.25%) and the LC50 values was 606.50 ppm for H. armigera and 1192.55 ppm for S. litura. The survived larvae produced malformed adults.