Plant-based hydrocarbon esters from Tragia involucrata possess antimicrobial and anti-inflammatory activities.

Antimicrobial and anti-inflammatory activities of hydrocarbon esters obtained from Tragia involucrata were evaluated by disk-diffusion (250 µg/ml), and broth-dilution (500-7.8 µg/ml), methods against bacteria. Among the compounds, shellsol showed the most potent activity against Burkholderia pseudomallei (KHW), Aeromonas hydrophila, Staphylococcus aureus, Bacillus subtilis, Streptococcus pyogenes, Klebsiella pneumoniae, Proteus mirabilis, and Streptococcus pneumoniae. Interestingly, vinyl hexylether was active against food-spoilage bacteria (Bacillus cereus and Proteus vulgaris), 2, 4-methyl hexane also exerted antimicrobial activity against K. pneumoniae, S. pyogenes, B. pseudomallei, Alcaligens viscolactis, and Pseudomonas aeruginosa. 2-methylnonane and 2, 6-dimethyl heptane showed only weak activity. For example, shellsol showed bacteriostatic effect (MIC of 7.8 µg/ml) against A. hydrophila, vinyl hexylether (MIC of 15.6 µg/ml) against P. mirabilis, and 2, 4-methyl hexane (MIC of 31.25 µg/ml) on B. pseudomallei. Cytotoxic effects of compounds were assayed in human skin and monkey kidney cells (62.5-2000 µg/ml) by an XTT assay. The vinyl hexylether, 2, 4-dimethyl hexane and shellsol did not show any toxicity up to 1000 µg/ml concentrations. The 2-methylnonane and 2, 6-dimethyl heptane induced morphological changes (e.g. cell disintegration and lysis) of both cell types at a 2000 µg/ml. The vinyl hexylether, 2, 4-dimethyl hexane and shellsol were devoid of toxic effects; however, 2-methylnonane induced weight loss and severe necrosis as evidenced by histopathological and serum biochemical analysis in rats. Interestingly, shellsol showed the maximum inhibition of carrageenan-induced, paw oedema in rats. In conclusion, findings of this study clearly indicate that biologically active hydrocarbon esters, such as shellsol, vinyl hexylether, and 2, 4-dimethyl hexane isolated from T. involucrata, may effectively control the growth of certain food-borne and food-spoilage pathogens.

Pilot Study with regard to the Wound Healing Activity of Protein from Calotropis procera (Ait.) R. Br.

We provide the scientific basis for the use of Calotropis procera for treating skin and wound infections in traditional medicine. The aqueous extract of stem-bark of C. procera exhibited more pronounced potent antimicrobial activity. Calo-protein was purified and identified from the most-active aqueous extracts of C. procera and showed broad-spectrum activity. Calo-protein inhibited the growth of S. aureus and E. aerogenes effectively at 25 µg/ml concentration. Mice topically treated with Calo-protein revealed significant wound healing after 14 days comparable to fusidic acid (FA) as positive control. This protein was devoid of cytolytic effect even at higher concentrations on skin cells after 24 h. Further investigation of this Calo-protein of C. procera on bacterial inhibition may provide a better understanding of the scientific basis and justification for its use in traditional medicine.

Electrochemical impedance spectroscopy characterization of nanoporous alumina dengue virus biosensor.

The Faradaic electrochemical impedance technique is employed to characterize the impedance change of a nanoporous alumina biosensor in response towards the specific binding of dengue serotype 2 (Denv2) viral particles to its serotype 2-specific immunoglobulin G antibody within the thin alumina layer. The optimal equivalent circuit model that matches the impedimetric responses of the sensor describes three distinct regions: the electrolyte solution (R(s)), the porous alumina channels (including biomaterials) (Q(1), R(1)) and the conductive electrode substrate layer (Q(2), R(2)). Both channel resistance R(1) and capacitance Q(1) change in response to the increase of the Denv2 virus concentration. A linear relationship between R(1) and Denv2 concentration from 1 to 900 plaque forming unit per mL (pfu mL(-1)) can be derived using Langmuir-Freundlich isotherm model. At 1pfu mL(-1) Denv2 concentration, R(1) can be distinguished from that of the cell culture control sample. Moreover, Q(1) doubles when Denv2 is added but remains unchanged in the presence of two other non-specific viruses - West Nile virus and Chikungunya virus indicates biosensor specificity can be quantitatively measured using channel capacitance.

Evaluation of antibacterial activity of proteins and peptides using a specific animal model for wound healing.

Wound healing is a complex process involving the integrated actions of numerous cell types, soluble mediators, and extracellular matrix (ECM). In this study, phospholipase A(2) (PLA(2)) purified from crotalid snake venom was found to express in vitro bactericidal activity against a group of clinical human pathogens. Based on the sequence homology of PLA(2), a series of peptides were derived from the C-terminal region of crotalid PLA(2). These short synthetic peptides were found to reproduce the bactericidal activity of its parent molecule. In vitro assays for bactericidal and cytolytic activities of these peptides showed very high microbicidal potency against Gram-negative and Gram-positive (Staphylococcus aureus) bacteria. Variants of the peptides showed reduced toxicity toward normal human cells, while retaining high bactericidal potency. Here we describe the protocol for evaluating the wound healing process by antibacterial peptides. We evaluated the biological roles of the candidate peptides in skin wound healing, using a specific BALB/c mice model. Peptide-treated animals showed accelerated healing of full-thickness skin wounds, with increased reepithelialization, collagen synthesis, and angiogenesis observed during the healing process. Healing wounds in protein/peptide-treated mice had higher densities of neutrophils, macrophages, and fibrocytes. Along with increased leukocyte infiltration, levels of macrophage-derived chemokine expression were also upregulated. These results demonstrate that the protein/peptide derived from snake venoms promotes healing of skin wounds. The primary mechanism seems to be an increase in leukocyte infiltration, leading to locally elevated synthesis and release of collagen and growth factors.

Insights into antifolate activity of phytochemicals against Pseudomonas aeruginosa.

Pseudomonas aeruginosa is an opportunistic drug resistant pathogen. Drug interaction studies for phytochemicals (protocatechuic acid (PA), gallic acid (GA), quercetin (QUER), and myricetin (MYR)) in combination with antifolates (sulfamethoxazole (SMX) and trimethoprim (TMP)) are presented. Our results show that the combinations of SMX and phytochemicals are synergistic, whereas TMP in combination with phytochemicals results in additive mode of interaction. Molecular docking of phytochemicals in the active site of modeled P. aeruginosa dihydrofolate reductase (DHFR), an important enzyme in the folic acid biosynthesis pathway, shows that the phytochemicals QUER and MYR dock in the active site of P. aeruginosa DHFR with promoted binding at the NADP site, PA, and GA dock in the active site of P. aeruginosa DHFR with promoted binding at the folate binding site. Possible mode of action of these phytochemicals as anti-DHFR compounds in this bacterium is suggested. Taken together, the above findings provide novel insights to mode of interactions of these phytochemicals with antibiotics and may have significance as prospective leads in the development of antipseudomonal drug developments.

In vitro combinations of antibiotics and phytochemicals against Pseudomonas aeruginosa.

BACKGROUND AND PURPOSE: Antibiotic combinations are used to enhance antibacterial efficacy and to prevent the development of resistance. In this study, the in vitro activities of antibiotic and phytochemical combinations against Pseudomonas aeruginosa were tested by the fractional inhibitory concentration method, derived from the minimal inhibitory concentrations (MICs) of the agents in combination. METHODS: The antimicrobial activity of phytochemicals, alone and in combination with antibiotics, was evaluated using the checkerboard assay and time-kill curve methods. RESULTS: There was synergism between gentamicin and caffeic acid, and sulfadiazine and the 3 phytochemicals under investigation (protocatechuic acid, quercetin, caffeic acid). The MIC of sulfadiazine was 256 microg/mL, and of gentamicin was 2 microg/mL. When gentamicin was combined with one-quarter the MIC of caffeic acid, the MIC of gentamicin was reduced 4-fold. When sulfadiazine was tested with one-quarter the MIC of protocatechuic acid, quercetin, and caffeic acid, the MIC was reduced 4-fold in combination with each of the drugs. CONCLUSIONS: These results indicate the potential efficacy of phytochemicals in combination with antibiotics for enhancing total biological activity.