Control Efficacy of Entomopathogenic Fungus Purpureocillium lilacinum against Chili Thrips (Scirtothrips dorsalis) on Chili Plant.

In a laboratory assay, it was shown that B. bassiana BCC48145, BCC2660, and P. lilacinum TBRC10638 were the three strains that exhibited the highest insecticidal activity against chili thrips, causing 92.5% and 91.86% and 92.3% corrected mortality, respectively. The fungi B. bassiana BCC48145 and P. lilacinum TBRC10638 were selected for greenhouse spraying. Cytotoxicity test of the extracts from both fungi evaluated against 4 animal cell lines: KB; human oral cavity carcinoma, MCF7; human breast adenocarcinoma, NCI-H187; human small cell lung carcinoma and GFP-expressing Vero cells, showed none-cytotoxic to all cell lines. An efficacy validation in the greenhouse showed that P. lilacinum TBRC 10638 was more effective than B. bassiana BCC48145 and could control the thrips up to 80% when using the fungus at 108 spores/mL. The LC50 values of P. lilacinum TBRC 10638 against chili thrips based on total thrips count from two experiments were 1.42 × 108 and 1.12 × 107 spores/mL when the fungal spores were sprayed once a week. The optimal concentration of P. lilacinum TBRC 10638 spores for effective control of chili thrips was determined at 1.41 × 109 spores/mL. The average efficacy of P. lilacinum TBRC 10638 for thrips control from 3 field trials was 30.08%, 14.39%, and 29.92%. This result was not significantly different from that of the chemical insecticide treatment group, which showed efficacy at 19.27%, 14.92%, and 19.97%. Furthermore, there was no difference in productivity among the different treatment groups. Our results demonstrated that P. lilacinum TBRC 10638 is a promising biocontrol agent that could be used as an alternative to chemical insecticide for controlling chili thrips.

Application of film-forming solution as a transdermal delivery system of piperine-rich herbal mixture extract for anti-inflammation.

Piperine-rich herbal mixture (PHM) used in this study is a traditional Thai medicine that contains 21 oriental herbs. It is called "Sahastara remedy" and is officially included in the Thai National List of Essential Medicine since A.D. 2011. PHM has been used orally to relieve muscle and bone pains. It contains Piper nigrum fruits as a major constituent and also Piper retrofractum fruits, PHM thus has anti-inflammatory activities that mostly come from the bioactivities of piperine consisting of these pepper fruits. Unfortunately, PHM usually causes gastrointestinal side effects. Consequently, a topical product containing an alcoholic extract of PHM (PHM-E), i.e., film-forming solution (FFS) was developed to overcome this drawback. The aims of this study were to investigate the anti-inflammatory activity of PHM-E, to evaluate physicochemical properties and the anti-inflammatory activity of FFS containing PHM-E (PHM-E FFS). Anti-inflammatory activities of PHM-E were investigated in the RAW 264.7 cells. Physicochemical properties, in vitro toxicities and anti-inflammatory activities of PHM-E FFS including its dry film (PHM-E film) were determined. PHM-E showed anti-inflammatory activities with dose dependent manners via inhibition of nitric oxide and prostaglandin E2 production by the RAW 264.7 cells and promotion of the cell phenotype polarization from M1 to M2. PHM-E FFS had low viscosity and exhibited the Newtonian behavior. It provided elastic PHM-E film with low tensile strength. The release profile of piperine from PHM-E film followed a zero-kinetic model. PHM-E FFS demonstrated compatibility with the skin cells, minimal ocular irritant when accidentally splashing into the eye and moderate-to-high potency for inhibition of inflammatory symptoms in the rats. PHM-E FFS thus had potential for use in the further clinical study to investigate its efficacy and safety in patients.

Semisynthesis and antibacterial activities of nidulin derivatives.

Derivatives of the fungal depsidone, nidulin, have been synthesized in order to evaluate the potential of the chemical skeleton as antibacterial agents. Alkylation, acylation, and arylation reactions of nornidulin underwent in a regioselective manner to predominantly produce 8-O-substituted derivatives. Many of the semisynthetic derivatives showed more potent antibacterial activities than nidulin, In particular, 8-O-aryl ether derivatives displayed significant activities against Gram-positive bacteria, including Methicillin-resistant Staphylococcus aureus.

Gamma sterilization of diclofenac sodium loaded- N-trimethyl chitosan nanoparticles for ophthalmic use.

This study was conducted to investigate the effect of gamma irradiation on physicochemical properties of N-trimethyl chitosan (TMC), diclofenac sodium (DC) and diclofenac sodium loaded N-trimethylchitosan nanoparticles (DC-TMCNs), and to determine suitable doses of gamma rays for sterilization of DC-TMCNs. Physicochemical properties of TMC, DC and DC-TMCNs before and after exposure to gamma rays at various doses were investigated. It was found that gamma irradiation at doses of 5-25kGy did not cause any significant changes in physical and chemical properties of TMC, DC and DC-TMCNs. The bioburden of DC-TMCNs was 1.5×106 CFU/vial. The initial contaminating bacteria were radiosensitive bacteria. A number of microorganisms was reduced to 10-6 after exposure to 9.9kGy of gamma rays. Therefore, DC-TMCNs could be sterilized by gamma irradiation at a dose of 10kGy, which did not alter their physicochemical properties and did not produce any substances toxic to the eye.

Cadinane sesquiterpenoids from the basidiomycete Stereum cf. sanguinolentum BCC 22926.

Stereumins Q-U, together with known stereumins A, B, K, L, and N, as well as ent-strobilols E and G were isolated from the culture of Stereum cf. sanguinolentum BCC 22926. Their structures were elucidated by extensive spectroscopic analyses. The absolute configurations of stereumins A and Q, as well as ent-strobilol E were established by application of the modified Mosher's method. Stereumin T displayed antibacterial activity against Bacilluscereus with a MIC value of 3.97µM.

Physicochemical properties and anti-Propionibacterium acnes activity of film-forming solutions containing alpha-mangostin-rich extract.

The objective of this study was to study the effect of formulation compositions on physicochemical properties and anti-Propionibacterium acnes activity of film-forming solutions containing alpha-mangostin-rich extract (AM). Film-forming solution bases and film-forming solutions containing AM were prepared by using Eudragit RL PO or Klucel LF or combinations of them as film-forming polymers. Rheological properties, pH values of the solutions, and mechanical properties of the dry films were investigated. An optimized formulation was selected and evaluated for the film surface, in vitro AM release, an anti-P. acnes activity, and potential for being a skin irritant. It was found that mechanical properties of the dry films were affected by total polymer contents, ratios of Klucel LF/Eudragit RL PO, AM, and contents of triethyl citrate. The film-forming solutions containing AM had pH values around 7.0. Their flow curves exhibited Newtonian flow behaviors. The optimized formulation provided films possessing smooth and nonporous surfaces. These films showed greater anti-P. acnes activity than their base films without toxicity to skin fibroblasts. Furthermore, AM released from the film matrix obeyed Higuchi's equation. In conclusion, the film-forming solutions containing AM had potential for treatment of acne vulgaris caused by P. acnes. However, further in vivo study is necessary to determine their efficacy and safety for using in patients suffering from acne vulgaris.

Optimization and evaluation of thermoresponsive diclofenac sodium ophthalmic in situ gels.

This work was conducted to optimize and evaluate Pluronic F127-based thermoresponsive diclofenac sodium ophthalmic in situ gels (DS in situ gel). They were prepared by cold method and investigated their physicochemical properties i.e., pH, flow ability, sol-gel transition temperature, gelling capacity and rheological properties. An optimized formulation was selected and investigated its physicochemical properties before and after autoclaving, eye irritation potency in SIRC cells and rabbits. In vivo ophthalmic absorption was performed in rabbits. It was found that physicochemical properties of DS in situ gels were affected by formulation compositions. Increment of Pluronic F127 content decreased sol-gel transition temperature of the products while increase in Pluronic F68 concentration tended to increase sol-gel transition temperature. In this study, Carbopol 940 did not affect sol-gel transition temperature but it affected transparency, pH, and gelling capacity of the products. The optimized formulation exhibited sol-gel transition at 32.6 ± 1.1 °C with pseudoplastic flow behavior. It was lost diclofenac sodium content during autoclaving. However, it was accepted as safe for ophthalmic use and could increase diclofenac sodium bioavailability in aqueous humor significantly. In conclusion, the optimized DS in situ gel had potential for using as an alternative to the conventional diclofenac sodium eye drop. However, autoclaving was not a suitable sterilization method for this product.

Sesquiterpene lactone 12,8-eudesmanolides from the fungus Xylaria ianthinovelutina.

The investigation of Xylaria ianthinovelutina (Mont.) led to the isolation of four new 12,8-eudesmanolides (1-4). Their structures were determined by spectroscopy and the relative stereochemistry was assigned based on NOESY and CD spectral data. Compounds 1-4 exhibited biological activity against Plasmodium falciparum K-1 strain, NCI-H187, KB, MCF-7 with IC(50) values varying in the range 0.78-19.15 microg/mL.

Cytotoxicity against insect cells of entomopathogenic fungi of the genera Hypocrella (anamorph Aschersonia): possible agents for biological control.

Extracts from entomopathogenic fungi of the genus Hypocrella (7 species) and its anamorph Aschersonia (11 species) were screened for cytotoxicity to Sf9 and C6/36 insect cells and L929, BHK(21)C13 and HepG2 mammalian cells. Cytotoxic extracts to insect cells (ID50's < or = 10 microg ml(-1)) but not to mammalian cells (ID50's > or = 10 microg ml(-1)) conformed to the criteria of the project and were considered 'lead' extracts for further investigation. 'Leads' were found in two of the Hypocrella species: H. discoidea, and H. tamurai and in three of the Aschersonia species: A. samoensis, A. badia, and A. tamurai. Bioassay-guided fractionation of the cell extract of the fungus A. samoensis BCC 1393 led to the identification of two known anthraquinone dimers, (+)rugulosin (1) and skyrin (2) which showed selective toxicity towards insect cells. (+)Rugulosin (1) and skyrin (2) exhibited strong cytotoxic activity against the insect cell line Sf9 with respective ID50 values of 1.2 and 9.6 microg ml(-1), but showed weak activity toward mammalian cells. This first report of (+)rugulosin (1) and skyrin (2) in A. samoensis is confirmed and demonstrated in another four strains of A. samoensis isolated in Thailand. The preferential cytotoxicity against Sf9 insect cells gives evidence that these insect-pathogenic fungi of the Hypocrella/Aschersonia group might be useful as an agent for pest control.