Laccase mediated diclofenac transformation and cytotoxicity assessment on mouse fibroblast 3T3-L1 preadipocytes

Diclofenac is recently considered as one of the most devastating environmental pollutants, because of its biomagnification in the food chain which leads to potential harmful effects on non-targeted organisms. This study describes the optimized laccase mediated diclofenac transformation using response surface methodology (RSM) and cytotoxicity testing on mouse fibroblast 3T3-L1 preadipocytes. Three factors (laccase, syringaldehyde, reaction time) were used to optimize the diclofenac transformation. The optimum level of laccase, syringaldehyde and reaction time was found to be 1.91 U mL -1, 187 μg and 51 min for diclofenac transformation (20 mg L -1). The cytotoxicity assessment on mouse fibroblast 3T3-L1 preadipocytes showed that a maximum of 67.9% cell death occurred at 72 h treatment with diclofenac (200 μg mL-1), while the cells treated with laccase treated diclofenac (LTD) showed less toxicity on the cells. These findings can be addressed for the removal of diclofenac toxicit

Molecular identification, phylogeny and antifungal susceptibilities of dematiaceous fungi isolated from human keratomycosis

Purpose: To investigate the dematiaceous fungal profile of patients with ocular mycoses attending a tertiary eye care hospital in Coimbatore, India Methods: The identification of dematiaceous fungus based on their morphology, their genotypes, and the measurement of the minimum inhibitory concentrations (MICs) using microdilution method of routinely used antifungal drugs were all compared. Results: A total of 148 dematiaceous fungi were isolated during a study period of 27 months. Isolates were confirmed as Curvularia spp. (n = 98), Exserohilum spp. (n = 32), Alternaria spp. (n = 14), Exophiala spp. (n = 2), Cladosporium sp. (n = 1) and Aureobasidium sp. (n = 1). Out of 50 well grown isolates characterized genotypically based on the amplification and sequencing of the ITS region of the ribosomal RNA gene cluster and subsequent BLAST analysis, Curvularia lunata (n = 24), C. aeria (n = 1), C. spicifera (n = 8), C. hawaiiensis (n = 1), C. maydis (n = 2), C. papendorfii (n = 2), C. geniculata (n = 3), C. tetramera (n = 2) and Exs. rostratum (n = 7) were identified. In vitro antifungal susceptibilities of the most tested dematiaceous isolates showed that voriconazole had a MIC50 of 0.25 μg ml−1, while amphotericin B had a MIC50 of 0.25 μg ml−1 for Curvularia spp. and Alternaria spp. Conclusion: Voriconazole proved to be the most effective drug against the pigmented filamentous fungi, followed by amphotericin B, itraconazole and econazole