Development of fluorescence-based methods for non-invasive measurement of integrin microclustering

In order to better understand the cell signaling process, non-invasive methods to measure the integrin microclustering and examine the factors affecting the dynamics of these clusters are needed. This dissertation is focused on the development of non-invasive analytical tools to investigate the role of cytoplasmic proteins and cholesterol on the nanoscale dynamic localization of integrins using a technique called FRET, fluorescence resonance energy transfer. FRET is used to detect the proximity of molecules at nanometer distances (1-10 nm) and can be applied in diverse ways with various microscope configurations. FRET images obtained with such setups can be used to quantify the interactions between biomolecules within cells.</p

Development of fluorescence-based methods for non-invasive measurement of integrin microclustering

In order to better understand the cell signaling process, non-invasive methods to measure the integrin microclustering and examine the factors affecting the dynamics of these clusters are needed. This dissertation is focused on the development of non-invasive analytical tools to investigate the role of cytoplasmic proteins and cholesterol on the nanoscale dynamic localization of integrins using a technique called FRET, fluorescence resonance energy transfer. FRET is used to detect the proximity of molecules at nanometer distances (1-10 nm) and can be applied in diverse ways with various microscope configurations. FRET images obtained with such setups can be used to quantify the interactions between biomolecules within cells

MJaya-ELM: A Jaya algorithm with mutation and extreme learning machine based approach for sensorineural hearing loss detection

Sensorineural hearing loss (SNHL) is a common hearing disorder or deafness which accounts for about 90% of the reported hearing loss. Magnetic resonance imaging (MRI) has been found to be an effective neuroimaging technique for detecting SNHL. However, manual detection methods, mainly based on the visual inspection of MRI, are cumbersome, time-consuming and need skilled supervision. Hence, there is a great need to design a computer-aided detection system for fast, accurate and automated detection of SNHL. This paper presents a new method for automated diagnosis of SNHL through brain MR images. Fast discrete curvelet transform is employed for image decomposition. The features are extracted from various decomposed subbands at different scales and orientations. A set of discriminant features is then derived using PCA+LDA algorithm. A hybrid classifier is suggested by integrating extreme learning machine and Jaya optimization with mutation (MJaya-ELM) to distinguish hearing loss images from healthy MR images. The proposed hybrid method overcomes the drawbacks of traditional ELM and other learning algorithms for single layer feedforward neural network. The concept of mutation is introduced to conventional Jaya optimization (MJaya) for improving the global search ability of the solutions by providing additional diversity. The proposed system is evaluated on a well-studied database. The comparison results demonstrate that the proposed scheme outperforms the existing schemes in terms of overall accuracy and sensitivity over different classes. The effectiveness of the proposed MJaya-ELM algorithm is also compared with its counterparts such as PSO-ELM, DE-ELM, and Jaya-ELM, and the results indicate the superiority of MJaya-ELM

A comparative therapeutic evaluation of topical Ivermectin Vs topical Permethrin for the management of scabies

Objective: To compare the therapeutic efficacy of topical ivermectin 0.5% vs. topical permethrin 5% cream in the treatment of scabies. Methods: This was open labelled, parallel group, prospective and comparative clinical study. Depending on the treatment to be received, there were two study groups comprising 50 patients each. Group 1 patients received topical 5 % Permethrin cream on day 1 and repeat application on 1st week follow up and if required then on second week; Group 2 patients received topical 0.5% Ivermectin cream on day 1 and repeat application on 1st week follow up and if required then on second week. The patient were also given antipruritic drug Tablet levocetirizine in a dose of 5 mg once daily given for 2 to 3 weeks. A total of 50 patients in each group were included in the study. Results: More than half of the patients in both Group 1 (56%) and Group 2 (54%) were below 30 years. More than half of the patients in both Group 1 (54%) and Group 2 (52%) were males. Majority of site of lesions were absent at 2 weeks in both the groups. Moderate and severe itching of lesion became nil at 2 weeks in both the groups. Grade 2 of itching of lesion was in 2% at 2 weeks in both the groups. KOH was positive in 94% patients in Group 1 and in 96% in Group 2 at baseline which decreased to 14% in Group 1 and 18% in Group 2 at 1 week. KOH positivity became nil at 2 weeks in both the groups. Conclusion: Topical Permethrin 5 % and Ivermectin 0.5 % were equally effective in the treatment of scabies up to 2 weeks

Noninvasive Measurements of Integrin Microclustering under Altered Membrane Cholesterol Levels

Reported herein is a method that can be used to study the role of cholesterol in the microclustering of a ubiquitous class of membrane receptors, termed integrins. Integrin microclustering was measured using a fluorescence resonance energy transfer assay that does not require direct attachment of fluorescent donors or acceptors onto the integrins, and thus minimizes unwanted perturbations to integrin clustering. Membrane cholesterol levels were reduced using methyl-β-cyclodextrin (mβCD), as confirmed by Amplex Red assays of total cellular lipid or plasma membrane lipid extract. Subsequent changes in integrin microclustering were measured in cells expressing wild-type (WT) or mutant integrins. Although less integrin microclustering was measured after 27% membrane cholesterol depletion in a cell line expressing WT integrins, there was no statistically significant change for cells expressing α-cytoplasmic integrin mutants after a 45% reduction in plasma membrane cholesterol, and a significant increase in clustering for cells expressing ligand-binding domain integrin mutants after a 57% decrease in membrane cholesterol. These results are explained by differences in WT and mutant integrin partitioning into lipid nanodomains. Restoration of original cholesterol levels was used to confirm that the measured changes in membrane properties were cholesterol-dependent. No correlations between lipid diffusion and integrin microclustering were measured by means of fluorescence recovery after photobleaching using a fluorescent lipid mimetic. Similar lipid diffusion coefficients were measured after cholesterol depletion, irrespective of the integrins being expressed