24052464
Chemically induced unfolding of bovine serum albumin by urea and sodium dodecyl sulfate: a spectral study with the polarity-sensitive charge-transfer fluorescent probe (E)-3-(4-methylaminophenyl)acrylic acid methyl ester.
Sensitivity of the charge-transfer (CT) band of the fluorescence probe (E)-3-(4-methylaminophenyl)acrylic acid methyl ester (MAPAME) towards the polarity of its immediate environment is employed to investigate the binding interaction of the probe with bovine serum albumin (BSA) and uncoiling of BSA by the denaturants urea and sodium dodecyl sulfate micelles. Binding of the probe with BSA produces a blue shift and enhanced intensity of the CT emission band which clearly point toward a decrease in polarity of the immediate environment of MAPAME. This is expected, since binding with BSA moves the probe from a polar water environment to a much less polar, hydrophobic protein interior, where the CT band is expected to be blue-shifted. Higher intensity arises due to fewer non-radiative decay paths available to the probe in the hydrophobic protein environment. Chemically induced unfolding of BSA by urea and sodium dodecyl sulfate is tracked by monitoring the induced spectral changes of the protein-bound probe MAPAME. Red-edge excitation shift or REES, fluorescence resonance energy transfer (FRET) and anisotropy measurements are used to investigate and monitor these binding and unfolding processes.
26613213
Fluorescent Probing of Protein Bovine Serum Albumin Stability and Denaturation Using Polarity Sensitive Spectral Response of a Charge Transfer Probe
The polarity sensitive photo-induced intra-molecular charge transfer (ICT) fluorescence probe (E)-3-(4-methylamino-phenyl)-acrylic acid ethyl ester (MAPAEE) has been used to study the model protein Bovine Serum Albumin (BSA) in its native and thermal and urea induced denatured states. The interaction between BSA and the regular surfactant Sodium Dodecyl Sulphate (SDS) as well as the biologically relevant steroid-based amphiphile Sodium Deoxycholate (NaDC) has also been very keenly followed using this ICT probe. The variation of micellar properties of both SDS and NaDC with increasing ionic strengths and in presence of the chaotrope urea has also been well documemted by the same probe. Steady-state spectroscopy, FRET, and fluorescence anisotropy measurements have been used to gain better insight into these processes and the molecule MAPAEE to be a full-bodied fluorescent probe for studying such intricate biological systems, their properties and interactions.
40815524
Constrained photophysics of partially and fully encapsulated charge transfer probe (E)-3-(4-Methylaminophenyl) acrylic acid methyl ester inside cyclodextrin nano-cavities: evidence of cyclodextrins cavity dependent complex stoichiometry.
The polarity sensitive intra-molecular charge transfer (ICT) emission from (E)-3-(4-Methylaminophenyl) acrylic acid methyl ester (MAPAME) is found to show distinct changes once introduced into the nano-cavities of cyclodextrins in aqueous environment. Movement of the molecule from the more polar aqueous environment to the less polar, hydrophobic cyclodextrin interior is marked by the blue shift of the CT emission band with simultaneous fluorescence intensity enhancement. The emission spectral changes on complexation with the α- and β-CD show different stoichiometries as observed from the Benesi-Hildebrand plots. Fluorescence anisotropy and lifetime measurements were performed to probe the different behaviors of MAPAME in aqueous α- and β-CD solutions.
21780213
Interfacial and self-aggregation of binary mixtures of anionic and nonionic amphiphiles in aqueous medium.
Interfacial and bulk properties of sodium dodecyl sulfate (SDS), polyoxyethylene (20) sorbitan monooleate (Tween-80), and their binary mixtures in aqueous medium have been investigated using surface tension, conductance, and fluorescence measurements at different temperatures. The critical micelle concentration (CMC), counterion binding, polarity index, aggregation number, thermodynamics of interfacial adsorption, and micellization are evaluated. The entropic contribution is found to be the predominant factor in micelle formation. The aggregation number and the polarity index of the micelle interior estimated by the pyrene fluorescence technique show interesting temperature dependence. The size and composition of the mixed micelles vary with the total concentration. The micellar compositions, activities of the individual components in the mixed species, and intersurfactant interactions are also estimated following regular solution theory, excess thermodynamic functions, and a molecular thermodynamic model. The surfactant mixtures are found to be nonideal with a lower degree of counterion binding compared to the individual members. Mixed micelles are nonspherical in nature at all temperatures when the mole fraction (α) of the ionic part in mixtures is high but transform to spherical when α(SDS) is ≤0.50.
24502512
Conformational study of papain in the presence of sodium dodecyl sulfate in aqueous medium.
The interactions between a globular protein, papain and the anionic surfactant, sodium dodecyl sulfate (SDS) have been investigated in aqueous medium using fluorimetric, circular dichroism, Fourier transform infra-red, UV-vis spectrophotometric, dynamic light scattering, and nuclear magnetic resonance techniques. The conformational change of papain in aqueous solution has been studied in the presence of SDS. The results show the high alpha-helical content and unfolded structure of papain in the presence of SDS due to strong electrostatic repulsion leading to a "necklace and bead model" in protein-surfactant complexes.
46163931
Diagnostic accuracy in retinoblastoma.
This study was done to ascertain the accuracy of diagnosis and possible risk of misdiagnosis in suspected retinoblastoma cases by different pre-operative diagnostic procedures like fundoscopy, ultrasonography, computerised tomography, and aqueous cytology study. Twenty-four eyes excised with diagnosis of suspected retinoblastoma were included in the study. Final diagnosis was reached by histopathological examination of excised eyes and compared with pre-operative diagnosis by different diagnostic procedures and accuracy of each procedure was calculated. Twenty-two cases were finally diagnosed as retinoblastoma and 2 cases (8.33%) as other than retinoblastoma. Computerised tomography and aqueous humour cytology were two most specific (100%) tests. Computerised tomography was the most sensitive (95.4%) test. Aqueous cytology was poorly sensitive (9.09%). Both ultrasonography and fundoscopy demonstrated sensitivity over 90% but with low specificity (50%). When test results of fundoscopy and ultrasonography were combined, specificity of this combination was 100% and sensitivity was 95.4%, which match exactly the values of computerised tomography. Combination of fundoscopy and ultrasonography has accuracy as high as that of computerised tomography. In patients with clear media this combination can be used as an effective alternative to computerised tomography to reach a diagnosis with comparable accuracy.
45407832
How Do Homozygous Parents Affect TDT as a Test for Association?
The traditional transmission disequilibrium test (TDT) (Spielman et al., 1993) is a powerful test for association only in the presence of linkage. Since allele transmissions from homozygous parents do not carry any information on linkage, the TDT statistic uses data only on heterozygous parents. However, homozygous parents carry information on association between alleles at a marker locus and a disease locus. In this article, we explore whether inclusion of homozygous parents increases the power to detect association. The resultant test statistic follows a χ2 distribution with 2 degrees of freedom. Monte-Carlo simulations are included to compare the performance of this test with the traditional TDT under different disease models.
33844160
Detection of a large number of subgroup 1 human rotaviruses with a “long” RNA electropherotype
SummaryThe long or short electrophoretic migration patterns of group A human rotaviruses are linked to their subgroup antigenic specificities. Long pattern isolates usually belong to subgroup 2 (SG2) and short pattern to subgroup 1 (SG1). To date detection of only 4 isolates which do not follow this linkage, have been reported. In the present communication we report the detection of unusually large number (39 isolates) of long pattern human isolates with SG1 specificities.