Thermodynamics of monosaccharide binding to concanavalin A, pea (Pisum sativum) lectin, and lentil (Lens culinaris) lectin

Titration calorimetry measurements of the binding of methyl alpha-D-mannopyranoside (Me alpha Man), D-mannopyranoside (Man), methyl alpha-D-glucopyranoside (Me alpha Glu), and D-glucopyranoside (Glu) to concanavalin A (Con A), pea lectin, and lentil lectin were performed at 281 and 292 K in 0.01 M dimethylglutaric acid-NaOH buffer (pH 6.9) containing 0.15 M NaCl and Mn+2 and Ca+2 ions. The site binding enthalpies, delta H, are the same at both temperatures and range from -28.4 +/- 0.9 (Me alpha Man) to -16.6 +/- 0.5 kJ mol-1 (Glu) for Con A, from -26.2 +/- 1.1 (Me alpha Man) to -12.8 +/- 0.4 kJ mol-1 (Me alpha Glu) for pea lectin, and from -16.6 +/- 0.7 (Me alpha Man) to -8.0 +/- 0.2 kJ mol-1 (Me alpha Glu) for lentil lectin. The site binding constants range from 17 +/- 1 x 10(3) M-1 (Me alpha Man to Con A at 281.2 K) to 230 +/- 20 M-1 (Glu to lentil lectin at 292.6 K) and exhibit high specificity for Con A where they are in the Me alpha Man:Man:Me alpha Glu:Glu ratio of 21:4:5:1, while the corresponding ratio is 5:2:1.5:1 for pea lectin and 4:2:2:1 for lentil lectin. The higher specificity for Con A indicates more interactions between the amino acid residues at the binding site and the carbohydrate ligand than for the pea and lentil lectin-carbohydrate complexes. The carbohydrate-lectin binding results exhibit enthalpy-entropy compensation in that delta Hb (kJ mol-1) = -1.67 +/- 0.06 x 10(4) + (1.30 +/- 0.12)T(K) delta Sb (J mol-1K-1). Differential scanning calorimetry measurements on the thermal denaturation of the lectins and their carbohydrate complexes show that the Con A tetramer dissociates into monomers, while the pea and lentil lectin dimers dissociate into two submonomer fragments. At the denaturation temperature, one carbohydrate binds to each monomer of Con A and the pea and lentil lectins. Complexation with the carbohydrate increases the denaturation temperature of the lectin and the magnitude of the increases yield binding constants in agreement with the determinations from titration calorimetry.

Topography of the combining region of a Thomsen-Friedenreich-antigen-specific lectin jacalin (Artocarpus integrifolia agglutinin). A thermodynamic and circular-dichroism spectroscopic study

Thermodynamic analysis of carbohydrate binding by Artocarpus integrifolia (jackfruit) agglutinin (jacalin) shows that, among monosaccharides, Me alpha GalNAc (methyl-alpha-N-acetylgalactosamine) is the strongest binding ligand. Despite its strong affinity for Me alpha GalNAc and Me alpha Gal, the lectin binds very poorly when Gal and GalNAc are in alpha-linkage with other sugars such as in A- and B-blood-group trisaccharides, Gal alpha 1-3Gal and Gal alpha 1-4Gal. These binding properties are explained by considering the thermodynamic parameters in conjunction with the minimum energy conformations of these sugars. It binds to Gal beta 1-3GalNAc alpha Me with 2800-fold stronger affinity over Gal beta 1-3GalNAc beta Me. It does not bind to asialo-GM1 (monosialoganglioside) oligosaccharide. Moreover, it binds to Gal beta 1-3GalNAc alpha Ser, the authentic T (Thomsen-Friedenreich)-antigen, with about 2.5-fold greater affinity as compared with Gal beta 1-3GalNAc. Asialoglycophorin A was found to be about 169,333 times stronger an inhibitor than Gal beta 1-3GalNAc. The present study thus reveals the exquisite specificity of A. integrifolia lectin for the T-antigen. Appreciable binding of disaccharides Glc beta 1-3GalNAc and GlcNAc beta 1-3Gal and the very poor binding of beta-linked disaccharides, which instead of Gal and GalNAc contain other sugars at the reducing end, underscore the important contribution made by Gal and GalNAc at the reducing end for recognition by the lectin. The ligand-structure-dependent alterations of the c.d. spectrum in the tertiary structural region of the protein allows the placement of various sugar units in the combining region of the lectin. These studies suggest that the primary subsite (subsite A) can accommodate only Gal or GalNAc or alpha-linked Gal or GalNAc, whereas the secondary subsite (subsite B) can associate either with GalNAc beta Me or Gal beta Me. Considering these factors a likely arrangement for various disaccharides in the binding site of the lectin is proposed. Its exquisite specificity for the authentic T-antigen, Gal beta 1-3GalNAc alpha Ser, together with its virtual non-binding to A- and B-blood-group antigens, Gal beta 1-3GalNAc beta Me and asialo-GM1 should make A. integrifolia lectin a valuable probe for monitoring the expression of T-antigen on cell surfaces.

Charge distribution in and electric moments of some aliphatic acids

Using the treatment of Smith et al., charge distribution in and consequently the dipole moments of several aliphatic acids have been evaluated. The electric moments of chloro (2·86 D), bromo (2·90 D), iodo (2·06 D) and trichloro (3·00 D) acetic acids have been measured in dioxan solution at 35°. The experimental values are compared with those calculated theoretically and discussed in terms of the various possible structures.

The 5-Methyl Group in Thymine Dynamically Influences the Structure of A-Tracts in DNA at the Local and Global Level

DNA sequences containing a stretch of several A:T basepairs without a 5'-TA-3' step are known as A-tracts and have been the subject of extensive investigation because of their unique structural features such as a narrow minor groove and their crucial role in several biological processes. One of the aspects under investigation has been the influence of the 5-methyl group of thymine on the properties of A-tracts. Detailed molecular dynamics simulation studies of the sequences d(CGCAAAUUUGCG) and d(CGCAAATTTGCG) indicate that the presence of the 5-methyl group in thymine increases the frequency of a narrow minor groove conformation, which could facilitate its specific recognition by proteins, and reduce its susceptibility to cleavage by DNase I. The bias toward a wider minor groove in the absence of the thymine 5-methyl group is a static structural feature. Our results also indicate that the presence of the thymine 5-methyl group is necessary for calibrating the backbone conformation and the basepair and dinucleotide step geometry of the core A-tract as well as the flanking CA/TG and the neighboring GC/GC steps, as observed in free and protein-bound DNA. As a consequence, it also fine-tunes the curvature of the longer DNA fragment in which the A-tract is embedded.

The ratio of diffusion coefficient to mobility for slow electrons in dry air

Using Huxley's solution of the diffusion equation for electron-attaching gases, the ratio of diffusion coefficient D to mobility μ for electrons in dry air was measured over the range 3·06 × 10-17 d strength (V cm-1) and N the gas density (cm -3). The results agreed with those of earlier workers who used the diffusion equation for non-attaching gases.

Studies on removal of metal ions and sulphate reduction using rice husk and Desulfotomaculum nigrificans with reference to remediation of acid mine drainage

The utility of rice husk as an adsorbent for metal ions such as iron, zinc and copper from acid mine water was assessed. The adsorption isotherms exhibited Langmuirian behavior and were endothermic in nature. The free energy values for adsorption of the chosen metal ions onto rice husk were found to be highly negative attesting to favorable interaction. Over 99% Fe3+, 98% of Fe2+ and Zn2+ and 95% Cu2+ uptake was achieved from acid mine water, with a concomitant increase in the pH value by two units using rice husk. The remediation studies carried out on acid mine water and simulated acid mine water pretreated with rice husk indicated successful growth of Desulfotomaculum nigrificans (D. nigrificans). The amount of sulphate bioreduction in acid mine water at an initial pH of 5.3 was enhanced by D. nigrificans from 21% to 40% in the presence of rice husk filtrate supplemented with carbon and nitrogen. In simulated acid mine water with fortified husk filtrate, the sulphate reduction was even more extensive, with an enhancement to 73%. Concurrently, almost 90% Fe2+, 89% Zn2+ and 75% Cu2+ bioremoval was attained from simulated acid mine water. Metal adsorption by rice husk was confirmed in desorption experiments in which almost complete removal of metal ions from the rice husk was achieved after two elutions using 1 M HCl. The possible mechanisms of metal ion adsorption onto rice husk and sulphate reduction using D. nigrificans are discussed.

Effects of Parasitics and Interface Traps on Ballistic Nanowire FET in the Ultimate Quantum Capacitance Limit

In this paper, we focus on the performance of a nanowire field-effect transistor in the ultimate quantum capacitance limit (UQCL) (where only one subband is occupied) in the presence of interface traps (D-it), parasitic capacitance (C-L), and source/drain series resistance (R-s,R-d), using a ballistic transport model and compare the performance with its classical capacitance limit (CCL) counterpart. We discuss four different aspects relevant to the present scenario, namely: 1) gate capacitance; 2) drain-current saturation; 3) subthreshold slope; and 4) scaling performance. To gain physical insights into these effects, we also develop a set of semianalytical equations. The key observations are as follows: 1) A strongly energy-quantized nanowire shows nonmonotonic multiple-peak C-V characteristics due to discrete contributions from individual subbands; 2) the ballistic drain current saturates better in the UQCL than in the CCL, both in the presence and absence of D-it and R-s,R-d; 3) the subthreshold slope does not suffer any relative degradation in the UQCL compared to the CCL, even with Dit and R-s,R-d; 4) the UQCL scaling outperforms the CCL in the ideal condition; and 5) the UQCL scaling is more immune to R-s,R-d, but the presence of D-it and C-L significantly degrades the scaling advantages in the UQCL.

Control of morphogenesis in tobacco protoplast cultures: organogenesis vs embryogenesis

The morphogenetic pathway leading to plant differentiation in tobacco mesophyll protoplasts could be regulated. The course of development via organogenesis or embryogenesis was controlled by manipulating nutrient media, culture conditions and hormone requirements. A lowering of molarity of medium after 5 weeks of protoplast culture, inclusion of GA3 (0.5 mg/l) in the medium for first 8 weeks of culture and exclusion of reduced nitrogen in the medium resulted in shoot organogenesis, while maintenance of higher molarity of the medium till 8 weeks, reduced nitrogen in the medium and removal of 2, 4-D after 5 weeks of culture induced embryogenesis. Regenerability of viable plants was obtained by both developmental pathways. The implications of tobacco embryogenesis system in plant molecular genetics were highlighted.

Probing the role of the C-H center dot center dot center dot O hydrogen bond stabilized polypeptide chain reversal at the C-terminus of designed peptide helices. Structural characterization of three decapeptides

The structural characterization in crystals of three designed decapeptides containing a double D-segment at the C-terminus is described. The crystal structures of the peptides Boc-Leu-Aib-Val-Xxx-Leu-Aib-Val- (D)Ala-(D)Leu-Aib-OMe, (Xxx = Gly 2, (D)Ala 3, Aib 4) have been determined and compared with those reported earlier for peptide 1 (Xxx = Ala) and the all L analogue Boc-Leu-Aib-Val-Ala-Leu-Aib-Val-Ala-Leu-Aib-OMe, which yielded a perfect right-handed a-helical structure. Peptides 1 and 2 reveal a right-handed helical segment spanning residues 1 to 7, ending in a Schellman motif with Ala(8) functioning as the terminating residue. Polypeptide chain reversal occurs at residue 9, a novel feature that appears to be the consequence of a C-(HO)-O-... hydrogen bond between residue 4 (CH)-H-alpha and residue 9 CO groups. The structures of peptides 3 and 4, which lack the pro R hydrogen at the C-alpha atom of residue 4, are dramatically different. Peptide 3 adopts a right-handed helical conformation over the 1 to 7 segment. Residues 8 and 9 adopt at conformations forming a C-terminus type I' beta-turn, corresponding to an incipient left-handed twist of the polypeptide chain. In peptide 4, helix termination occurs at Aib(6), with residues 6 to 9 forming a left-handed helix, resulting in a structure that accommodates direct fusion of two helical segments of opposite twist. Peptides 3 and 4 provide examples of chiral residues occurring in the less favored sense of helical twist; (D)Ala(4) in peptide 3 adopts an alpha(R) conformation, while (L)Val(7) in 4 adopts an alpha(L) conformation. The structural comparison of the decapeptides reported here provides evidence for the role of specific C-(HO)-O-... hydrogen bonds in stabilizing chain reversals at helix termini, which may be relevant in aligning contiguous helical and strand segments in polypeptide structures.

Interactions of methoxyamine with pyridoxal-5'-phosphate-Schiff's base at the active site of sheep liver serine hydroxymethyltransferase

The mechanism of interaction of methoxyamine with sheep liver serine hydroxymethyltransferase (EC 2.1.2.1) (SHMT) was established by measuring changes in enzyme activity, visible absorption spectra, circular dichroism and fluorescence, and by evaluating the rate constant by stopped-flow spectrophotometry. Methoxyamine can be considered as the smallest substituted aminooxy derivative of hydroxylamine. It was a reversible noncompetitive inhibitor (Ki = 25 microM) of SHMT similar to O-amino-D-serine. Like in the interaction of O-amino-D-serine and aminooxyacetic acid, the first step in the reaction was very fast. This was evident by the rapid disappearance of the enzyme-Schiff base absorbance at 425 nm with a rate constant of 1.3 x 10(3) M-1 sec-1 and CD intensity at 430 nm. Concomitantly, there was an increase in absorbance at 388 nm (intermediate I). The next step in the reaction was the unimolecular conversion (1.1 x 10(-3) sec-1) of this intermediate to the final oxime absorbing at 325 nm. The identity of the oxime was established by its characteristic fluorescence emission at 460 nm when excited at 360 nm and by high performance liquid chromatography. These results highlight the specificity in interactions of aminooxy compounds with sheep liver serine hydroxymethyltransferase and that the carboxyl group of the inhibitors enhances the rate of the initial interaction with the enzyme.

Detection of edges from projections

In a number of applications of computerized tomography, the ultimate goal is to detect and characterize objects within a cross section. Detection of edges of different contrast regions yields the required information. The problem of detecting edges from projection data is addressed. It is shown that the class of linear edge detection operators used on images can be used for detection of edges directly from projection data. This not only reduces the computational burden but also avoids the difficulties of postprocessing a reconstructed image. This is accomplished by a convolution backprojection operation. For example, with the Marr-Hildreth edge detection operator, the filtering function that is to be used on the projection data is the Radon transform of the Laplacian of the 2-D Gaussian function which is combined with the reconstruction filter. Simulation results showing the efficacy of the proposed method and a comparison with edges detected from the reconstructed image are presented

Correlation of resistivity and microstructure in dilute titaniulll–nitrogen alloy

The precipitation kinetics of the quenched dilute Ti-1.6 at.-%N alloy has been followed by resistivity measurements at 77 K using the four probe method. Resistivity behaviour has been studied for various durations for the alloys aged in the temperature range 273-373 K. The resistivity behaviour has been explained on the basis of the growth and decay of interfacial strain fields associated with the precipitation process. In addition, the resistivity changes have been correlated with transmission electron microscopy observations. (C) 1995 The Institute of Materials.

Sensing of delaminations in composite laminates using embedded magnetostrictive particle layers

This is an exploratory study to illustrate the feasibility of detecting delamination type of damage in polymeric laminates with one layer of magnetostrictive particles. One such beam encircled with excitation and sensing coils is used for this study. The change in stress gradient of the magnetostrictive layer in the vicinity of delamination shows up as a change in induced voltage in the sensing coil, and therefore provides a means to sense the presence of delamination. Recognizing the constitutive behavior of the Terfenol-D material is highly nonlinear, analytical expressions for the constitutive relations are developed by using curve fitting techniques to the experimental data. Analytical expressions that relate the applied excitation field with the stress and magnetic flux densities induced in the magnetostrictive layer are developed. Numerical methods are used to find the relative change in the induced voltage in the sensing coil due to the presence of delamination. A typical example of unidirectional laminate, with embedded delaminations, is used for the simulation purposes. This exploratory study illustrates that the open-circuit voltage induced in the sensing coil changes significantly (as large of 68 millivolts) with the occurrence of delamination. This feature can be exploited for device off-line inspection techniques and/or linking monitoring procedures for practical applications.

Effect of two-level systems on the spectral density of universal conductance fluctuations in doped Si

We have studied the power spectral density [S(f) = gamma/f(alpha)] of universal conductance fluctuations (UCF's) in heavily doped single crystals of Si, when the scatterers themselves act as the primary source of dephasing. We observed that the scatterers, with internal dynamics like two-level-systems, produce a significant, temperature-dependent reduction in the spectral slope alpha when T less than or similar to 10 K, as compared to the bare 1/f (alphaapproximate to1) spectrum at higher temperatures. It is further shown that an upper cutoff frequency (f(m)) in the UCF spectrum is necessary in order to restrict the magnitude of conductance fluctuations, [(deltaG(phi))(2)], per phase coherent region (L-phi(3)) to [(deltaGphi)(2)](1/2) less than or similar to e(2)/h. We find that f(m) approximate to tau(D)(-1), where tau(D) = L-2/D, is the time scale of the diffusive motion of the electron along the active length (L) of the sample (D is the electron diffusivity).

Longwave radiative forcing of Indian Ocean tropospheric aerosol

A spectrally resolved discrete-ordinates radiative transfer model is used to calculate the change in downwelling surface and top-of-the-atmosphere (TOA) outgoing longwave (3.9-500 mum) radiative fluxes induced by tropospheric aerosols of the type observed over the Indian Ocean during the Indian Ocean Experiment (INDOEX). Both external and internal aerosol mixtures were considered. Throughout the longwave, the aerosol volume extinction depends more strongly on relative humidity than in most of the shortwave (0.28-3.9 mum), implying that particle growth factors and realistic relative humidity profiles must be taken into account when modeling the longwave radiative effects of aerosols. A typical boundary layer aerosol loading, with a 500-nm optical depth of 0.3, will increase the downwelling longwave flux at the surface by 7.7 W m(-2) over the clean air case while decreasing the outgoing longwave radiation by 1.3 W m(-2). A more vertically extended aerosol loading, exhibiting a high opacity plume between 2 and 3 km above the surface and having a typical 500-nm optical depth of 0.7, will increase the downwelling longwave flux at the surface by 11.2 W m(-2) over the clean air case while decreasing the outgoing longwave radiation by 2.7 W m(-2). For a vertically extended aerosol profile, approximately 30% of the TOA radiative forcing comes from sea salt and approximately 60% of the forcing comes from the combination of sea salt and dust. The remaining forcing is from anthropogenic constituents. These results are for the external mixture. For an internal mixture, TOA longwave forcings can be up to a factor of two larger. Therefore, to complete our understanding of this region's longwave aerosol radiative properties, more detailed information is needed about aerosol mixing states. These longwave radiative effects partially offset the large shortwave aerosol radiative forcing and should be included in regional and global climate modeling simulations.