A novel method for mapping assembled epitopes in batches: Identification of three epitopes at the receptor binding region of human chorionic gonadotropin

Identification of conformation-specific epitopes of hCG beta has been done using a simple batch method, Chemically or enzymatically-modified hCG beta has been prepared in a batch and the effect of modifications on the integrity of different epitope regions has been investigated in a quantitative manner using monoclonal antibodies (MAbs) immobilized on plastic tubes from culture supernatants. Based on the extent of damage done to different regions by different modifications, three conformation-specific epitopes of hCG beta have been identified. The method has been shown to have important advantages over the existing methods on many considerations, Using this approach, these epitopes have been shown to be at/near the receptor-binding region.

Electron transfer reaction in the marcus inverted region: role of high frequency vibrational modes

A theoretical study of the dynamics of photo-electron transfer reactions in the Marcus inverted regime is presented. This study is motivated partly by the recent proposal of Barbara et al. (J. Phys. Chem. 96, 3728, 1991) that a minimal model of an electron transfer reaction should consist of a polar solvent mode (X), a low frequency vibrational mode (Q) and one high frequency mode (q). Interplay between these modes may be responsible for the crossover observed in the dynamics from a solvent controlled to a vibrational controlled electron transfer. The following results have been obtained. (i) In the case of slowly relaxing solvents, the proximity of the point of excitation to an effective sink on the excited surface is critical in determining the decay of the reactant population. This is because the Franck-Condon overlap between the reactant ground and the product excited states decreases rapidly with increase in the quantum number of the product vibrational state. (ii) Non-exponential solvation dynamics has an important effect in determining the rates of electron transfer. Especially, a biphasic solvation and a large coupling between the reactant and the product states both may be needed to explain the experimental results. ©1996 American Institute of Physics

Quantum and non-Markovian effects in the electron transfer reaction dynamics in the Marcus inverted region

Electron transfer reactions in large molecules may often be coupled to both the polar solvent modes and the intramolecular vibrational modes of the molecule. This can give rise to a complex dynamics which may in some systems, like betaine, be controlled more by vibrational rather than by solvent effects. Additionally, a significant contribution from an ultrafast relaxation component in the solvation dynamics may enhance the complexity. To explain the wide range of behavior that has been observed experimentally, Barbara et al. recently proposed that a model of an electron transfer reaction should minimally consist of a low-frequency classical solvent mode (X), a low-frequency vibrational mode (Q), and a high-frequency quantum mode (q) (J. Phys. Chem. 1991, 96, 3728). In the present work, a theoretical study of this model is described. This study generalizes earlier work by including the biphasic solvent response and the dynamics of the low-frequency vibrational mode in the presence of a delocalized, extended reaction zone. A novel Green's function technique has been developed which allowed us to study the non-Markovian dynamics on a multidimensional surface. The contributions from the high-frequency vibrational mode and the ultrafast component in the non-Markovian solvent dynamics are found to be primarily responsible for the dramatic increase in charge transfer rate over the prediction of the classical theories that neglect both these factors. These, along with a large coupling between the reactant and the product states, may combine to render the electron transfer rate both very large and constant over a wide range of solvent relaxation rates. A study on the free energy gap dependence of the electron transfer rate reveals that the rates are sensitive to changes in the quantum frequency particularly when the free energy gap is very large.

Status Survey of the Malabar Pied Hornbill in the Dandeli Region, Northern Western Ghats, India

The Malabar Pied Hornbill, Anthracoceros coronatus, is a near threatened species, endemic to the tropical deciduous forests of central and southern India and Sri Lanka. The Dandeli region in Karnataka (India) is believed to be the last stronghold of this species in the Western Ghats biodiversity hotspot. Being a rapidly developing area with a growing human population, the threats to this species and their habitat are mounting, especially due to a large number of hydroelectric projects and habitat fragmentation caused by paper and plywood industries. This study evaluated the change in population status of the Malabar Pied Hornbill over a 23 year period and defined priorities for the long term conservation and monitoring of hornbills in Dandeli. Encounter rates of hornbills were also analysed in relation to the density and species richness of trees and fruiting trees, basal area, canopy cover and distance from river. Hornbill encounters were not significantly different compared to the earlier study carried out by Reddy in 1988, but were significantly different across the five sites in the current study. Higher numbers of hornbills were encountered closer to the river, but these results were only marginally significant. The mean numbers of hornbills recorded at the two roost sites identified in Dandeli were 26 +/- 4.47 (n=16 counts) and 31.78 +/- 3.53 (n=14 counts) respectively. The study also helped build local awareness about the species, train local Forest Department staff in monitoring hornbills and develop a management plan for its conservation.

Implications of the recent high statistics determination of the pion electromagnetic form factor in the timelike region

The recently evaluated two-pion contribution to the muon g - 2 and the phase of the pion electromagnetic form factor in the elastic region, known from pi pi scattering by Fermi-Watson theorem, are exploited by analytic techniques for finding correlations between the coefficients of the Taylor expansion at t = 0 and the values of the form factor at several points in the spacelike region. We do not use specific parametrizations, and the results are fully independent of the unknown phase in the inelastic region. Using for instance, from recent determinations, < r(pi)(2)> = (0.435 +/- 0.005) fm(2) and F(-1.6 GeV2) = 0.243(-0.014)(+0.022), we obtain the allowed ranges 3.75 GeV-4 less than or similar to c less than or similar to 3.98 GeV-4 and 9.91 GeV-6 less than or similar to d less than or similar to 10.46 GeV-6 for the curvature and the next Taylor coefficient, with a strong correlation between them. We also predict a large region in the complex plane where the form factor cannot have zeros.

Thiol/disulphide exchange reaction: A key regulatory process in biological systems

Cysteine residues in proteins serve many important functions such as stabilizing and maintaining the three-dimensional conformation of many proteins(1), in enzyme catalysis, as a residue undergoing post-translational 2 and in the formation of DNA-binding modification domain of a class of transcriptional activators(3), It is also involved in biological redox coupling(4) and xenobiotic metabolism(5). Disulphide bonds formed by xenobiotic metabolism oxidation of cysteine residues have been used as a probe to study the structure/function relationships of proteins, Introducing novel disulphide bonds in proteins to increase their thermal stability and, therefore, the shelf life is an important goal of protein engineering(6,7), In addition, the thiol group of cysteine residue participates in a reaction termed as thiol/disulphide exchange reaction, the biological significance of this reaction being the theme of this review.

Turbulent heat flux variation over the Monsoon-Trough region during MONTBLEX-90

Tower data collected during the Monsoon-Trough Boundary Layer Experiment (MONTBLEX-90) have been analysed to understand the observed structure of the surface layer over an arid region (Jodhpur) and a moist region (Kharagpur) during active and weak phases of the 1990 southwest monsoon. Turbulent heat and momentum fluxes are estimated by the eddy correlation method using sonic data. The turbulent momentum flux at both Jodhpur and Kharagpur was larger when the winds were stronger, reaching a maximum of the order of 0.5 N m(-2) on 5 and 6 August when a low pressure system was located over the region. The heat flux at Jodhpur is high during weak monsoon days, the maximum being 450 W m(-2), whereas during active days the flux never exceeds 200 W m(-2). At Kharagpur, the flux does not vary significantly between active and weak monsoon days, the maximum in either phase being 160 W m(-2) At Jodhpur, there is significant contrast in the near-surface air temperature, being higher during weak monsoon days as compared to active days. Cloud cover did not vary significantly in both the regions. The turbulent heat flux variation at both the sites appears to be correlated mainly with soil mixture, and less sensitive to cloud cover.

Isolation of an 800 kb contiguous DNA fragment encompassing a 3.5-cM region of chromosome 1 in Arabidopsis using YAC clones

The apetalal mutation of Arabidopsis affects floral meristem identity and the development of sepal and petal primordia of the flower. We mapped the available RFLP markers on chromosome 1 that are in the general vicinity of apetalal on a fine structure map and then chose the closest RFLP as a starting point for contiguous DNA (contig) generation. We report here a contig of about 800 kilobases (kb) that spans a 3.5 cM region of chromosome 1. We used genomic libraries of Arabidopsis prepared in yeast artificial chromosome (YAC) vectors and the detailed characterization of 19 YACs is reported. RFLPs displayed by the end fragments from the walk were mapped to align and correlate the genetic and physical maps for this region of chromosome 1. In this segment of the genome, 1 cM corresponds to a little over 200 kb of physical distance.

Unsteady compressible boundary layer flow in the stagnation region of a sphere with a magnetic field

Abstract: An analysis is performed to study the unsteady compressible laminar boundary layer flow in the forward stagnation-point region of a sphere with a magnetic field applied normal, to the surface. We have considered the case where there is an initial steady state that is perturbed by the step change in the total enthalpy at the wall. The nonlinear coupled parabolic partial differential equations governing the flow and heat transfer have been solved numerically using a finite-difference scheme. The numerical results are presented, which show the temporal development of the boundary layer. The magnetic field in the presence of variable electrical conductivity causes an overshoot in the velocity profile. Also, when the total enthalpy at the wall is suddenly increased, there is a change in the direction of transfer of heat in a small interval of time.

Localized data gathering paradigms for small and marginal farm lands in Semi-Arid region – Issues and Concerns

In this work we explore the application of wireless sensor technologies for the benefit of small and marginal farmers in semi-arid regions. The focus in this paper is to discuss the merits and demerits of data gathering & relay paradigms that collect localized data over a wide area. The data gathered includes soil moisture, temperature, pressure, rain data and humidity. The challenge to technology intervention comes mainly due to two reasons: (a) Farmers in general are interested in crop yield specific to their piece of land. This is because soil texture can vary rapidly over small regions. (b) Due to a high run-off, the soil moisture retention can vary from region to region depending on the topology of the farm. Both these reasons alter the needs drastically. Additionally, small and marginal farms can be sandwiched between rich farm lands. The village has very little access to grid power. Power cuts can extend up to 12 hours in a day and upto 3 or 4 days during some months in the year. In this paper, we discuss 3 technology paradigms for data relaying. These include Wi-Fi (Wireless Fidelity), GPRS (General Packet Radio Service) and DTN (Delay and Disruption Tolerant Network) technologies. We detail the merits and demerits of each of these solutions and provide our final recommendations. The project site is a village called Chennakesavapura in the state of Karnataka, India.

A fixed-grid finite element based enthalpy formulation for generalized phase change problems: role of superficial mushy region

The enthalpy method is primarily developed for studying phase change in a multicomponent material, characterized by a continuous liquid volume fraction (phi(1)) vs temperature (T) relationship. Using the Galerkin finite element method we obtain solutions to the enthalpy formulation for phase change in 1D slabs of pure material, by assuming a superficial phase change region (linear (phi(1) vs T) around the discontinuity at the melting point. Errors between the computed and analytical solutions are evaluated for the fluxes at, and positions of, the freezing front, for different widths of the superficial phase change region and spatial discretizations with linear and quadratic basis functions. For Stefan number (St) varying between 0.1 and 10 the method is relatively insensitive to spatial discretization and widths of the superficial phase change region. Greater sensitivity is observed at St = 0.01, where the variation in the enthalpy is large. In general the width of the superficial phase change region should span at least 2-3 Gauss quadrature points for the enthalpy to be computed accurately. The method is applied to study conventional melting of slabs of frozen brine and ice. Regardless of the forms for the phi(1) vs T relationships, the thawing times were found to scale as the square of the slab thickness. The ability of the method to efficiently capture multiple thawing fronts which may originate at any spatial location within the sample, is illustrated with the microwave thawing of slabs and 2D cylinders. (C) 2002 Elsevier Science Ltd. All rights reserved.

Insights into the Regulatory Characteristics of the Mycobacterial Dephosphocoenzyme A Kinase: Implications for the Universal CoA Biosynthesis Pathway

Being vastly different from the human counterpart, we suggest that the last enzyme of the Mycobacterium tuberculosis Coenzyme A biosynthetic pathway, dephosphocoenzyme A kinase (CoaE) could be a good anti-tubercular target. Here we describe detailed investigations into the regulatory features of the enzyme, affected via two mechanisms. Enzymatic activity is regulated by CTP which strongly binds the enzyme at a site overlapping that of the leading substrate, dephosphocoenzyme A (DCoA), thereby obscuring the binding site and limiting catalysis. The organism has evolved a second layer of regulation by employing a dynamic equilibrium between the trimeric and monomeric forms of CoaE as a means of regulating the effective concentration of active enzyme. We show that the monomer is the active form of the enzyme and the interplay between the regulator, CTP and the substrate, DCoA, affects enzymatic activity. Detailed kinetic data have been corroborated by size exclusion chromatography, dynamic light scattering, glutaraldehyde crosslinking, limited proteolysis and fluorescence investigations on the enzyme all of which corroborate the effects of the ligands on the enzyme oligomeric status and activity. Cysteine mutagenesis and the effects of reducing agents on mycobacterial CoaE oligomerization further validate that the latter is not cysteine-mediated or reduction-sensitive. These studies thus shed light on the novel regulatory features employed to regulate metabolite flow through the last step of a critical biosynthetic pathway by keeping the latter catalytically dormant till the need arises, the transition to the active form affected by a delicate crosstalk between an essential cellular metabolite (CTP) and the precursor to the pathway end-product (DCoA).

Phenobarbitone-mediated translocation of the cytosolic proteins interacting with the 5 '-proximal region of rat liver CYP2B1/B2 gene into the nucleus

The positive element (PE) (-69 to -98 bp) within the 5'-proximal region of the CYP2B1B2 gene (+1 to -179 bp) of rat liver is essential for phenobarbitone (PB) response and gives a single major complex with the rat liver cytosol in gel shift analysis. This complex corresponds to complex I (top) of the three complexes given by the nuclear extracts. PB treatment of rats leads to a decrease in complex I formation with the cytosol and PE and an increase in the same with the nuclear extract in gel shift analysis. Both the changes are counteracted by simultaneous okadaic acid administration. The nuclear protein giving rise to complex I has been isolated and has an M-r of 26 kDa. The cytosolic counterpart consists of two species, 26 and 28 kDa, as revealed by Southwestern blot analysis using labeled PE. It is concluded that PB treatment leads to the translocation accompanied by processing of the cytosolic protein species into the nucleus that requires protein dephosphorylation. It is suggested that PB may exert a global regulation on the transcription of many genes by modulating the phosphorylation status of different protein factors involved in transcriptional regulation. (C) 2002 Elsevier Science (USA).

Unsymmetrical aryl disulfides with excellent transparency in the visible region for second order nonlinear optics

Several unsymmetrically substituted aromatic donor acceptor disulfides have been synthesized and analysed for their second order nonlinear optical properties. These molecules exhibit moderately high first hyperpolarizability (beta) with excellent transparency in the visible region. Most of the unsymmetrical disulfides have a cut-off wavelength below 420 nm. Calculations show that the molecules have an asymmetric charge distribution around the disulfide bond which is responsible for their high beta values. These results provide motivation for the design and synthesis of nonlinear optical chromophores with multiple disulfide bonds for large second order nonlinearity and excellent visible transparency.