ABSTRACT
BACKGROUND: Tuberculosis (TB) caused 1.8 million deaths worldwide with increased multiple drug resistance (MDR) cases estimated 4.8 lakhs in the year 2015. ß-Lactam antibiotics could be a hope for TB treatment. Therefore, in this study, uniformity in the biochemical and molecular nature of ß-lactamases was analyzed to evaluate the potential of ß-lactam antibiotics as a treatment regimen against Mycobacterium tuberculosis (MTB). MATERIALS AND METHODS: ß-Lactamase enzymes in 233 MTB clinical isolates along with control H37Rv strain were characterized by enzyme kinetic using nitrocefin and cefotaxime as a substrate, isoelectric points by isoelectric focusing electrophoresis (IEF) and by PCR and Southern blotting. RESULTS: Enzyme kinetics showed Km and Vmax for nitrocefin in the range of 56-69µM and 7.00-11IU/lit respectively, for cefotaxime in the range of 0.35-0.59µM and 18-25IU/lit respectively. ß-Lactamase showed high affinity for clavulanic acid an inhibitor of Extended-Spectrum ß-lactamase enzymes (ESBLs). The pIs of 4.9 and 5.1 were observed for all the MTB clinical isolates and control H37Rv. Southern blotting confirmed the presence of blaC sequence in MTB chromosomal DNA. CONCLUSION: This confirmed that MTB ß-lactamase enzymes belong to the Class A, group 2be Extended Spectrum ß-Lactamases with no biochemical or molecular polymorphism. ESBLs are mainly responsible for resistance against ß-lactam antibiotics in MTB. Thus ESBLs could be the potential therapeutic target for TB treatment using ß-lactam antibiotics in combination with ß-lactamase inhibitors like sulbactam and sodium clavulanate.
Subject(s)
Anti-Bacterial Agents , Mycobacterium tuberculosis , beta-Lactamases , beta-Lactams , Humans , Anti-Bacterial Agents/metabolism , beta-Lactam Resistance/genetics , beta-Lactamases/genetics , beta-Lactamases/metabolism , beta-Lactams/metabolism , Blotting, Southern , Cefotaxime/metabolism , Cephalosporins/metabolism , Clavulanic Acid/metabolism , Isoelectric Focusing , Kinetics , Mycobacterium tuberculosis/genetics , Mycobacterium tuberculosis/metabolism , Polymerase Chain Reaction , Tuberculosis/microbiologyABSTRACT
Diabetes mellitus (DM) is one of the most common lifestyle diseases, caused due to endocrine disorder. DM is commonly associated with hyperglycemia, a condition which is generally followed by an overproduction of free radicals leading to tissue oxidative stress. Currently, the focus of medical fraternity lies in developing therapeutic drugs based on natural origin in order to reduce the hyperglycemia associated toxicity. Ferulic acid (FA) is a ubiquitous hydroxycinnamic acid displaying an array of therapeutic properties, including anti-diabetic effect which could be attributed to its potent antioxidant capacity. However, due to low bioavailability and clinical efficacy of FA, its biomedical applications remained limited. In the present study, FA encapsulated chitosan nanoparticles (FANPs) were synthesized through ionotropic gelation process with an aim to enhance FA bioavailability. The plasma release and urinary excretion profiles of FANPs were compared with that of free FA using healthy Wistar albino rats as a model system. The encapsulated FA displayed extended plasma retention time and maximum plasma concentration was recorded at 60â¯min which implied four times enhancement of Tmax compared to free FA. The elimination of compound from the animal body also displayed a similar pattern where the peak urinary excretion of FA from nanoformulations. FANPs were also tested for their anti-hyperglycemic effects in streptozotocin (STZ) induced diabetes in Wistar albino rats and were found to attenuate the diabetes-associated symptoms. FANPs caused an enhancement in body weight, decrease in blood glucose level along with a regulatory effect on blood lipid profile of diabetic rats. Positive impact of FANPs in improving the hyperglycemic condition prevalent in diabetic rats might provide new avenues for the treatment of DM and help avoid secondary complications associated with the synthetic drugs.
Subject(s)
Coumaric Acids/therapeutic use , Diabetes Mellitus, Experimental/drug therapy , Nanoparticles/chemistry , Administration, Oral , Animals , Blood Glucose/metabolism , Calorimetry, Differential Scanning , Coumaric Acids/blood , Coumaric Acids/pharmacokinetics , Delayed-Action Preparations , Diabetes Mellitus, Experimental/blood , Hypoglycemic Agents/pharmacokinetics , Hypoglycemic Agents/therapeutic use , Lipids/blood , Nanoparticles/ultrastructure , Pancreas/pathology , Particle Size , Proton Magnetic Resonance Spectroscopy , Rats, Wistar , Spectroscopy, Fourier Transform Infrared , Static Electricity , Toxicity Tests, AcuteABSTRACT
Candida albicans, an opportunistic fungal pathogen is a major causative agent of superficial to systemic life-threating biofilm infections on indwelling medical devices. These biofilms acts as double edge swords owing to their resistance towards antibiotics and immunological barriers. To overcome this threat ferulic acid encapsulated chitosan nanoparticles (FA-CSNPs) were formulated to assess its efficacy as an antibiofilm agent against C. albicans. These FA-CSNPs were synthesized using ionotropic gelation method and observed through field emission scanning electron microscopy (FESEM) and fluorescent microscopy. Assessment of successful encapsulation and stability of ferulic acid into chitosan nanoparticles was made using Fourier transform infrared spectrum (FTIR), (1)H NMR and thermal analyses. Synthesized FA-CSNPs, were found to be cytocompatible, when tested using Human Embryonic Kidney (HEK-293) cell lines. XTT assay revealed that FA-CSNPs reduced the cell metabolic activity of C. albicans upto 22.5% as compared to native ferulic acid (63%) and unloaded CSNPs (88%) after 24 h incubation. Disruption of C. albicans biofilm architecture was visualized by FESEM. Results highlighted the potential of FA-CSNPs to be used as an effective alternative to the conventional antifungal therapeutics.
