Comprehensive Comparative Analysis of Cholesterol Catabolic Genes/Proteins in Mycobacterial Species.

In dealing with Mycobacterium tuberculosis, the causative agent of the deadliest human disease-tuberculosis (TB)-utilization of cholesterol as a carbon source indicates the possibility of using cholesterol catabolic genes/proteins as novel drug targets. However, studies on cholesterol catabolism in mycobacterial species are scarce, and the number of mycobacterial species utilizing cholesterol as a carbon source is unknown. The availability of a large number of mycobacterial species' genomic data affords an opportunity to explore and predict mycobacterial species' ability to utilize cholesterol employing in silico methods. In this study, comprehensive comparative analysis of cholesterol catabolic genes/proteins in 93 mycobacterial species was achieved by deducing a comprehensive cholesterol catabolic pathway, developing a software tool for extracting homologous protein data and using protein structure and functional data. Based on the presence of cholesterol catabolic homologous proteins proven or predicted to be either essential or specifically required for the growth of M. tuberculosis H37Rv on cholesterol, we predict that among 93 mycobacterial species, 51 species will be able to utilize cholesterol as a carbon source. This study's predictions need further experimental validation and the results should be taken as a source of information on cholesterol catabolism and genes/proteins involved in this process among mycobacterial species.

Encapsulation and stability of clofazimine liposomes.

The entrapment of clofazimine (CLO) in a liposomal delivery system for topical application can protect it from absorption into the blood circulation and increase its residence time within the skin. This may reduce the very long mean period of leprosy treatment, as well as the side effects due to the long term administration of large doses of the drug. This investigation deals with critical parameters controlling the formulation and stabilization of liposomes with encapsulated CLO. The entrapment efficiency of CLO in liposomes was increased by altering the proportion of phosphatidyl choline (PC) and cholesterol (CHOL) in liposomes. The stability of liposomal suspensions and the liposomal gels (HPMC K4M) in terms of retention of CLO was measured at refrigeration temperature (2-8 degrees C), room temperature (25 +/- 2 degrees C) and body temperature (37 degrees C) for a period of 3 months. The results show that entrapment of CLO in liposomes can be increased by increasing the proportion of PC. However, the optimum encapsulation and retention of CLO was achieved only with a specific PC:CHOL molar ratio (5.13:1.00). An almost identical value of the entrapment efficiency was obtained when gel filtration and ultracentrifugation methods were used to separate the CLO-carrying liposomes from free drug. The effect of vortexing and sonication on the entrapment efficiency gave similar results, although the mean particle size was different. CLO liposomal gels were found to be stable at room temperature for up to 3 months.