Cytotoxicity and Anti-Inflammatory Effects of Polyherbal Formulations, Joint Pain Spl and Rumalaya Forte on Lipopolysaccharide Induced Inflammation in IC-21 Macrophages.

AIM: To test the effectiveness of marketed polyherbal formulations on lipopolysaccharide-induced inflammatory conditions in macrophages. BACKGROUND: Usage of herbal compounds among patients suffering from arthritis and cancer is increasing every year. Many anti-inflammatory herbal products available in the market should be screened thoroughly for their possible mechanism of action. OBJECTIVE: Joint Pain Spl (JPS) is a polyherbal dietary food supplement composed of 13 herbal plants, and Rumalaya Forte (RF) is a polyherbal formulation comprising of 6 herbal plants. These were tested for their cytotoxicity, as well as antioxidant and anti-inflammatory activities in LPS treated IC-21 peritoneal macrophages. METHODS: Commercially available JPS and RF powder was used to prepare the extract. The aqueous and methanol extracts were quantified for the presence of phenolic and flavonoid compounds and confirmed with HPLC. In vitro DPPH free scavenging activity was performed. Cytotoxicity was tested by MTT assay. Anti-inflammatory activity was tested using lipopolysaccharide-stimulated IC-21 peritoneal macrophage cells. RESULTS: The phytochemical screening showed the presence of phenolic and flavonoid compounds in JPS and RF. The aqueous and methanol extracts of JPS and RF possesses significant DPPH free radical scavenging activity. MTT assay revealed that 90.64% (aqueous extract) and 92.21% (methanol extract) of exposed macrophages are viable even after 24h exposure of maximal tested concentrations of herbal formulations. Pre-treatment of JPS and RF on LPS induced IC-21 macrophages showed a reduction in nitric oxide production (maximal 79.95%) and a high level of superoxide anion scavenging activity (maximal 82.5%) over control. CONCLUSION: The two tested polyherbal formulations, such as JPS and RF possess anti-inflammatory activity by modulating free radical generation in IC-21 macrophages. Thus the presence of the phenolic and flavonoid compounds may contribute to the antioxidant activity.

Annona muricata: An alternate mosquito control agent with special reference to inhibition of detoxifying enzymes in Aedes aegypti.

This study was aimed to investigate an effectual level of Annona muricata (soursop) extracts on mosquito vectors namely, Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus. The toxicity study on non-target organism and other important biochemical marker enzymes to find and illustrate the exact mechanism of specific enzymes responsible for detoxifying allelochemicals. Among the various soursop seed kernel extracts tested for larvicidal activity, the 0.9% saline extract exhibited maximum mortality (100%) against three vectors at the lowest concentration for 24 h exposure. Based on these findings, the saline extract was opted for further studies including toxicity on non-target organism and systemic effects on important biochemical constituents in the larvae A. aegypti at the lethal threshold time (18 h) with LC50 concentration (0.009 mg/mL). The tested extract against non-target aquatic fourth instar larvae Chironomus costatus was safe up to 0.0028 mg/mL for 24 h exposure and the mortality was observed only above the concentration 0.0028 mg/mL used in the study. The systemic effects on main neuron transmitter Acetylcholinesterase (p ≤ 0.01), xenobiotics detoxifying enzyme of α-and ß-carboxylesterase (p ≤ 0.05; p ≤ 0.01) and antioxidant enzyme glutathione S-transferase (p ≤ 0.05) were reduced significantly in quantitative analysis. Analysis of such biochemical constituents of proteins and enzymes α-and ß-carboxylesterase were considerably down regulated in the resolving native-PAGE. In contrast, acid and alkaline phosphatase were upregulated in both quantitative and qualitative analysis. This investigation clearly demonstrates the soursop extract has potent larvicidal agent with alterations in biochemical constituents of exposed larvae of A. aegypti.

Green synthesis of silver-nanoparticles from Annona reticulata leaves aqueous extract and its mosquito larvicidal and anti-microbial activity on human pathogens.

Silver nanoparticles play a important role in controlling mosquito population as well as multi drug resistant pathogens without causing much harm to humans. In the present study was focused on green synthesis of silver nanoparticles against dengue causing vector (Aedes aegypti) and pathogens affecting humans. The synthesized silver nanoparticle was confirmed using UV- absorption spectrum range obtained at 416 nm, XRD, FTIR and HR-TEM analysis were used to determine the silver nanoparticle morphology and size with ∼6.48 ± 1.2-8.13 ± 0.18 nm and face centered cubic structure. The synthesized silver nanoparticles were exposed to fourth instar larvae of A. aegypti with different concentration (3-20 µg/mL) for 24 h and its elicit maximum mortality (100%) at their final concentration of 20 µg/mL and it's LC50 value was 4.43 µg/mL and LC90 value was 13.96 µg/mL, respectively. The minimum inhibitory activities of the tested pathogens were 125, 31.25, 62.5, 62.6 and 62.5 µg/mL for the Bacillus cereus, Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Candida albicans respectively. Further, the synthesized silver nanoparticle shows a potent antimicrobial activity against all tested pathogens. Moreover the effect of silver nanoparticle against Red Blood Cells belonging to 'O' positive blood group were tested and does not cause higher hemolysis to the cells even at the highest concentration. Based on these finding, we strongly suggested that face centered cubic structured A. reticulata AgNPs is an eco-friendly and potent bio-medical agent and can be apply in wide range of application an alternative chemically synthesized metal nanoparticle.