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.

title: 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.
abstract: 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.

title: The role of sodium dodecyl sulfate (SDS) micelles in inducing skin barrier perturbation in the presence of glycerol.
abstract: The stratum corneum (SC) serves as the skin barrier between the body and the environment. When the skin is contacted with an aqueous solution of the surfactant sodium dodecyl sulfate (SDS), a well-known model skin irritant, SDS penetrates into the skin and disrupts this barrier. It is well established, both in vitro and in vivo, that the SDS skin penetration is dose-dependent, and that it increases with an increase in the total SDS concentration above the critical micelle concentration (CMC) of SDS. However, when we added the humectant glycerol at a concentration of 10 wt% to the aqueous SDS contacting solution, we observed, through in vitro quantitative skin radioactivity assays using (14)C-radiolabeled SDS, that the dose dependence in SDS skin penetration is almost completely eliminated. To rationalize this important observation, which may also be related to the well-known beneficial effects of glycerol on skin barrier perturbation in vivo, we hypothesize that the addition of 10 wt% glycerol may hinder the ability of the SDS micelles to penetrate into the skin barrier through aqueous pores that exist in the SC. To test this hypothesis, we conducted mannitol skin permeability as well as average skin electrical resistivity measurements in vitro upon exposure of the skin to an aqueous SDS contacting solution and to an aqueous SDS + 10 wt% glycerol contacting solution in the context of a hindered-transport aqueous porous pathway model of the SC. Our in vitro studies demonstrated that the addition of 10 wt% glycerol: (i) reduces the average aqueous pore radius resulting from exposure of the skin to the aqueous SDS contacting solution from 33 +/- 5 Angstrom to 20 +/- 5 Angstrom, such that a SDS micelle of radius 18.5 +/- 1 Angstrom (as determined using dynamic light-scattering measurements) experiences significant steric hindrance and cannot penetrate into the SC, and (ii) reduces the number density of aqueous pores in the SC by more than 50%, thereby further reducing the ability of the SDS micelles to penetrate into the SC and perturb the skin barrier.

title: Evidence for interstate transmission and increase in prevalence of bovine group B rotavirus strains with a novel VP7 genotype among diarrhoeic calves in Eastern and Northern states of India.
abstract: During a surveillance study (2003-2005) in a cattle market in Kolkata city, state of West Bengal, Eastern India, 34 (13.0%) of 260 calves with diarrhoea were positive for group B rotaviruses (GBR) by RNA electrophoresis in polyacrylamide gels. Analysis of the partial VP7 gene sequence of 28 of the 34 GBR strains revealed maximum identities (97.7-99.5% at nucleotide level and 97.8-100% at amino-acid level) with the novel bovine GBR 'Kolkata strains' reported in an earlier surveillance study (1.5%, n=192, 2001-2002) from the same cattle market, and shared low identities of 73.7-78.9% and 80.8-89.6%; 62.6-66.2% and 59.8-65.4%; 58.9-62.2% and 48.6-54.9% at nucleotide and amino-acid level with other bovine, human, and murine GBR. The GBR-infected calves were traced to districts in neighbouring states of West Bengal. Therefore, the present study reports a rapid increase in prevalence (13.0% in 2003-2005 against 1.5% in 2001-2002) of novel GBR strains among calves with diarrhoea, and provides evidence for interstate transmission of GBR.

title: Induction maintenance concept for HAART as initial treatment in HIV infected infants
abstract: BackgroundEarly initiated antiretroviral therapy (ART) in HIV infected infants leads to improved long-term viral suppression and survival. Guidelines recommend initiating therapy with a triple ART consisting of two nucleoside reverse transcriptase inhibitors (NRTIs) and either one additional non-nucleoside reverse transcriptase inhibitor (NNRTI) or a protease inhibitor (PI). Compared to older children and adults, viral relapse is seen more frequently in infants receiving triple ART. We now address the possibility of a more potent ART with a quadruple induction and triple maintenance therapy.MethodsWe examine the longitudinal course in four HIV infected infants, who were referred from other centers and could not be recruited to multicentre trials. We introduced ART initially consisting of two NRTIs, one NNRTI and one PI and later discontinued the PI at the age of 12 months maintaining a triple regime consisting of two NRTIs and one NNRTI.ResultsProvided that therapy adherence was maintained we observed an effective sustained decline of viral load and significant CD4 cell reconstitution even after switching to a triple regime. No drug associated toxicity was seen.ConclusionWe suggest that a four drug therapy might be a possible initial therapy option in HIV infected infants, at least in those with a high viral load, followed by a maintenance triple regime after 12 months of therapy.

title: Why is sodium cocoyl isethionate (SCI) mild to the skin barrier? - An in vitro investigation based on the relative sizes of the SCI micelles and the skin aqueous pores.
abstract: Sodium cocoyl isethionate (SCI) is an important surfactant ingredient in mild, syndet (synthetic detergent) cleansing bars. In vitro and in vivo studies have demonstrated that SCI is mild and less damaging to the skin barrier than soaps and surfactants such as sodium dodecyl sulfate (SDS). We have recently shown that SDS forms small micelles in aqueous solutions contacting the skin relative to the aqueous pores in the stratum corneum (SC), and as a result, the SDS micelles can contribute to SDS skin penetration and induce skin barrier perturbation. In this paper, we attempt to explain the well-documented skin mildness of SCI by examining the size of the SCI micelles relative to that of the aqueous pores in the SC. For this purpose, we have conducted in vitro mannitol skin permeability and average skin electrical resistivity measurements upon exposure of the skin to an aqueous SCI contacting solution in the context of a hindered-transport aqueous porous pathway model of the SC. These in vitro studies demonstrate that an SCI micelle of radius 33.5 +/- 1 Angstrom (as determined using dynamic light-scattering measurements) experiences significant steric hindrance and cannot penetrate into the SC through aqueous pores that have an average radius of 29 +/- 5 Angstrom. We believe that this inability of the SCI micelles to contribute to SCI skin penetration and associated skin barrier perturbation is responsible for the observed skin mildness of SCI. Through in vitro quantitative skin radioactivity assays using (14)C-radiolabeled SCI and pig full-thickness skin (p-FTS), we also show conclusively that SCI skin penetration is dose-independent, an important finding that provides additional evidence that the larger SCI micelles cannot penetrate into the SC through the smaller aqueous pores that exist in the SC, and therefore, cannot induce skin barrier perturbation.

title: The Papworth BiVent tube: a feasibility study of a novel double-lumen endotracheal tube and bronchial blocker in human cadavers.
abstract: A novel double-lumen endotracheal tube, the Papworth BiVent tube, has been designed to allow the rapid passage of a blocker into either main bronchus, without fibreoptic endoscopic guidance.



The feasibility of lung isolation and one-lung ventilation (OLV) in human cadavers is examined, along with displacement of the bronchus blocker during head and neck movement.



Cadaveric endotracheal intubation with the Papworth BiVent tube was straightforward and comparable with intubation with a conventional single-lumen tube (SLT). Reliable lung isolation was achieved considerably faster using the Papworth BiVent tube than with a bronchoscopically guided bronchial blocker through an SLT (mean 7.75 s BiVent tube vs 128.2 s SLT). The Papworth BiVent tube also prevented displacement of the blocker from its position in the bronchus on head movement.



This study in human cadavers has shown that it is feasible to use the Papworth BiVent tube to attain rapid and secure lung isolation for OLV. Further work is required in clinical settings.