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1.
Benef Microbes ; 6(1): 129-39, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25213027

ABSTRACT

Increasing levels of antibiotic resistance by Staphylococcus aureus have posed a need to search for non-antibiotic alternatives. This study aimed to assess the inhibitory effects of crude and fractionated cell-free supernatants (CFS) of locally isolated lactic acid bacteria (LAB) against a clinical strain of S. aureus. A total of 42 LAB strains were isolated and identified from fresh vegetables, fresh fruits and fermented products prior to evaluation of inhibitory activities. CFS of LAB strains exhibiting a stronger inhibitive effect against S. aureus were fractionated into crude protein, polysaccharide and lipid fractions. Crude protein fractions showed greater inhibition against S. aureus compared to polysaccharide and lipid fractions, with a more prevalent effect from Lactobacillus plantarum 8513 and L. plantarum BT8513. Crude protein, polysaccharide and lipid fractions were also characterised with glycine, mannose and oleic acid being detected as the major component of each fraction, respectively. Scanning electron microscopy revealed roughed and wrinkled membrane morphology of S. aureus upon treatment with crude protein fractions of LAB, suggesting an inhibitory effect via the destruction of cellular membrane. This research illustrated the potential application of fractionated extracts from LAB to inhibit S. aureus for use in the food and health industry.


Subject(s)
Anti-Bacterial Agents/pharmacology , Complex Mixtures/pharmacology , Lactobacillus/chemistry , Staphylococcus aureus/drug effects , Staphylococcus aureus/growth & development , Anti-Bacterial Agents/isolation & purification , Cell Membrane/drug effects , Cell Membrane/ultrastructure , Chemical Fractionation , Complex Mixtures/isolation & purification , Lactobacillus/isolation & purification , Lipids/isolation & purification , Lipids/pharmacology , Microscopy, Electron, Scanning , Polysaccharides/isolation & purification , Polysaccharides/pharmacology , Proteins/isolation & purification , Proteins/pharmacology , Vegetables/microbiology
2.
Gene Ther ; 16(8): 963-72, 2009 Aug.
Article in English | MEDLINE | ID: mdl-19474811

ABSTRACT

Small interfering RNAs (siRNAs) can be designed to specifically and potently target and silence a mutant allele, with little or no effect on the corresponding wild-type allele expression, presenting an opportunity for therapeutic intervention. Although several siRNAs have entered clinical trials, the development of siRNA therapeutics as a new drug class will require the development of improved delivery technologies. In this study, a reporter mouse model (transgenic click beetle luciferase/humanized monster green fluorescent protein) was developed to enable the study of siRNA delivery to skin; in this transgenic mouse, green fluorescent protein reporter gene expression is confined to the epidermis. Intradermal injection of siRNAs targeting the reporter gene resulted in marked reduction of green fluorescent protein expression in the localized treatment areas as measured by histology, real-time quantitative polymerase chain reaction and intravital imaging using a dual-axes confocal fluorescence microscope. These results indicate that this transgenic mouse skin model, coupled with in vivo imaging, will be useful for development of efficient and 'patient-friendly' siRNA delivery techniques and should facilitate the translation of siRNA-based therapeutics to the clinic for treatment of skin disorders.


Subject(s)
Green Fluorescent Proteins/genetics , Keratinocytes/metabolism , Mice, Transgenic , RNA Interference , RNA, Small Interfering/administration & dosage , Skin/metabolism , Animals , Genes, Reporter , Humans , Luciferases/genetics , Mice , Models, Animal
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