Studies on an enzyme reacting with isoniazid fromMycobacterium tuberculosis H37Rv

Mycobacterium tuberculosis H37Rv possesses an enzyme (referred to as ‘Y enzyme’) which catalyses in the presence of INH and NAD, the formation of a product, which turns yellow on acidification. The requirements for the reaction, such as enzyme concentration, INH concentration, etc., have been standardized. The substrate specificity of the enzyme with respect to INH and NAD has been determined. The reaction is specific for the INH-sensitive strain and is totally absent in INH-resistant strains. Furthermore, the ‘Y enzyme’ shows some characteristic features of a peroxidase in its requirement for oxygen and sensitivity to inhibition by various reagents. The requirements of this enzyme which is involved in the action of isoniazid inM. tuberculosis H37Rv is described for the first time.

The dynamic stability of degenerate systems under parametric excitation

The parametric resonance in a system having two modes of the same frequency is studied. The simultaneous occurence of the instabilities of the first and second kind is examined, by using a generalized perturbation procedure. The region of instability in the first approximation is obtained by using the Sturm's theorem for the roots of a polynomial equation.

A Need for FSH in Maintaining Fertility of Adult Male Subhuman Primates

Adult fertile male bonnet monkeys (Macaca radiata) were continuously deprived of endogenous follicle stimulating hormone (FSH) support for 240 days by injecting them with 1 ml of characterized monkey antiserum to oFSH every 48 hr; control monkeys received during the same period normal monkey serum instead. Testicular function was assessed at periodic intervals by (a) carrying out differential counting of sperm in the ejaculate obtained and (b) determining the hyaluronidase activity as well as in vitro 3H thymidine incorporation into DNA of testicular tissue removed at biopsy. Both the quality (viability and motility) of the sperms voided and the total sperm counts showed marked decreases as a function of time of immunization, the first significant reduction being noted by 100 days. FSH deprivation affected both the biochemical parameters used to test testicular functionality they being reduced at ∼200 days by 50%-60%. The fertility of these monkeys was evaluated at periodic times after 90 days of treatment by means of mating studies. FSH deprivation had rendered the monkeys incapable of impregnating any of the females used. Testosterone and luteinizing hormone (LH) levels remained unchanged following FSH antiserum injection. With cessation of antiserum treatment testicular function and fertility were completely restored to normalcy, indicating that the observed effect was specifically due to FSH deprivation. This study thus provides conclusive evidence for the involvement of FSH in maintenance of testicular function and fertility in the adult male primate.

Ratio of Diffusion Coefficient to Mobility for Electrons in SF6-Air and Freon-Nitrogen Mixtures

The ratio of diffusion coefficient to mobility (D/¿) for electrons has been measured in SF6-air and freon-nitrogen mixtures for various concentrations of SF6 and freon in the mixtures over the range 140¿ E/p¿ 220 V.cm-1 - torr-1. In SF6-air mixtures, the values of D/¿ were always observed to lie intermediate between the values for the pure gases. However, in freon-nitrogen mixtures, with a small concentration (10 percent) of freon in the mixture, the values of D/¿ are found to lie above the boundaries determined by the pure gases. In this mixture, over the lower E/p range (140 to 190) the electrons appear to lose a large fraction of their energy by the excitation of the complex freon molecules, while at higher E/p values (200 to 240), the excitation and consequent deexcitation of nitrogen molecules and its metastables seem to cause an increased rate of ionization of freon molecules.

Ionization and breakdown in sf6-air and freon-nitrogen mixtures

The sparking potentials and swarm coefficients (ionization and attachment coefficients) have been measured in sulphurhexafluoride- air and freon-nitrogen mixtures over the range of 110 ¿ E/p ¿ 240 V cm-1 torr-l and gas pressures varying between 1 and 20 torr, at 20°C. Addition of strongly attaching salphur-hexafluoride and freon gases increased the sparking potentials and the rate of increase of the attachment coefficient with increasing percentage of the strongly attaching gases in the mixtures was much larger than the rate of change of the first ionization coefficient.

Comparison of Prediction Based LSF Quantization Methods using Split VQ

Further improvement in performance, to achieve near transparent quality LSF quantization, is shown to be possible by using a higher order two dimensional (2-D) prediction in the coefficient domain. The prediction is performed in a closed-loop manner so that the LSF reconstruction error is the same as the quantization error of the prediction residual. We show that an optimum 2-D predictor, exploiting both inter-frame and intra-frame correlations, performs better than existing predictive methods. Computationally efficient split vector quantization technique is used to implement the proposed 2-D prediction based method. We show further improvement in performance by using weighted Euclidean distance.

Kinetics of formalization of poly(vinyl acetate)

Kinetic information on the formation of poly(vinyl formal) by the reaction of poly(vinyl acetate) and formaldehyde in presence of aqueous acid has been derived from the spectroscopic analysis of polymer samples after different periods of reaction. The hydroxyl content of poly(vinyl formal) is found to be nearly independent of reaction time and only slightly affected by temperature while the fall of acetate content and the increase in formal content are most rapid in the initial period and are largely influenced by temperature. The rate expression formulated on the assumption that the formalization reaction is of first order with respect to both poly(vinyl acetate) and formaldehyde explains the observed variation of polymer composition with reaction time. The activation energy for the reaction is found to be 17.3 kcal/mol.

Synthesis, X-ray structure and properties of [Ru2O(MeCN)4(O2CMe)2(PPh3)2](ClO4)2

The diruthenium(III) complex [{(PPh3)(MeCN)2Ru}2(μ-O)(μ-O2CMe)2](ClO4)2 (1) has been prepared from Ru2O(O2CMe)4(PPh3)2, which is obtained from a reaction of Ru2Cl(O2CMe)4 and PPh3 in MeCN. The crystal structure of 1 was determined by X-ray studies and the complex has an {Ru2(μ-O)(μ-O2CMe)22+} core and the facial sites on each metal centre are occupied by two MeCN and one PPh3 ligands. The Ru—b. Ru and Ru—Ooxo distances and Ru—O—Ru angle are 3.240(1), 1.866(4) Å and 120.6(2)°, respectively. The cis and trans Ru—N distances in 1 are 2.040(6) and 2.116(5) Å, respectively. The visible spectral band in 1 is observed at 574 nm (var epsilon, 10,800 M−1 cm−1). The 1H NMR spectrum of the diamagnetic complex 1 in CD3CN is in agreement with the X-ray structure.

The alpha method for solving differential algebraic inequality (DAI) systems

This paper describes an algorithm for ``direct numerical integration'' of the initial value Differential-Algebraic Inequalities (DAI) in a time stepping fashion using a sequential quadratic programming (SQP) method solver for detecting and satisfying active path constraints at each time step. The activation of a path constraint generally increases the condition number of the active discretized differential algebraic equation's (DAE) Jacobian and this difficulty is addressed by a regularization property of the alpha method. The algorithm is locally stable when index 1 and index 2 active path constraints and bounds are active. Subject to available regularization it is seen to be stable for active index 3 active path constraints in the numerical examples. For the high index active path constraints, the algorithm uses a user-selectable parameter to perturb the smaller singular values of the Jacobian with a view to reducing the condition number so that the simulation can proceed. The algorithm can be used as a relatively cheaper estimation tool for trajectory and control planning and in the context of model predictive control solutions. It can also be used to generate initial guess values of optimization variables used as input to inequality path constrained dynamic optimization problems. The method is illustrated with examples from space vehicle trajectory and robot path planning.

Development of Very High Frequency Damped Sine Wave Generator

In order to protect the critical electronic equipment/system against damped sine transient currents induced into its cables due to transient electromagnetic fields, switching phenomena, platform resonances, etc. it is necessary to provide proper hardening. The hardness assurance provided can be evaluated as per the test CS 116 of MIL STD 461E/F in laboratory by generating & inducing the necessary damped sine currents into the cables of the Equipment Under Test (EUT). The need and the stringent requirements for building a damped sine wave current generator for generation of damped sine current transients of very high frequencies (30 MHz & 100 MHz) have been presented. A method using LC discharge for the generation has been considered in the development. This involves building of extremely low & nearly loss less inductors (about 5 nH & 14 nH) as well as a capacitor & a switch with much lower inductances. A technique for achieving this has been described. Two units (I No for 30 MHz. & 100 MHz each) have been built. Experiments to verify the output are being conducted.

Formation of linear plasmid multimers promoted by the phage lambda Red-system in lon mutants of Escherichia coli

We report here the formation of plasmid linear multimers promoted by the Red-system of phage lambda using a multicopy plasmid comprised of lambda red alpha and red beta genes, under the control of the lambda cI857 repressor. Our observations have revealed that the multimerization of plasmid DNA is dependent on the red beta and recA genes, suggesting a concerted role for these functions in the formation of plasmid multimers. The formation of multimers occurred in a recBCD+ sbcB+ xthA+ lon genetic background at a higher frequency than in the isogenic lon+ host cells. The multimers comprised tandem repeats of monomer plasmid DNA. Treatment of purified plasmid DNA with exonuclease III revealed the presence of free double-chain ends in the molecules. Determination of the size of multimeric DNA, by pulse field gel electrophoresis, revealed that the bulk of the DNA was in the range 50-240 kb, representing approximately 5-24 unit lengths of monomeric plasmid DNA. We provide a conceptual framework for Red-system-promoted formation and enhanced accumulation of plasmid linear multimers in lon mutants of E. coli.

Synthesis of liquid crystalline benzothiazole base derivatives: A study of their optical and electrical properties

Two new donor-acceptor type liquid crystalline semiconductors based on benzothiazole have been synthesized. Their structural, photophysical and electronic properties were investigated using X-ray diffraction, atomic force microscopy, cyclic voltammetry, UV-Vis, photoluminescence, and Raman spectroscopy. The liquid crystalline behaviour of the molecules was thoroughly examined by differential scanning calorimetry (DSC) and optical polarizing microscope. The DSC and thermogravimetric analysis (TGA) show that these materials posses excellent thermal stability and have decomposition temperatures in excess of 300 degrees C. Beyond 160 degrees C both molecules show a smectic A liquid crystalline phase that exists till about 240 degrees C. Field-effect transistors were fabricated by vacuum evaporating the semiconductor layer using standard bottom gate/top contact geometry. The devices exhibit p-channel behaviour with hole mobilities of 10(-2) cm(2)/Vs. (C) 2009 Elsevier B.V. All rights reserved.

Branch-branch connections in trees analogous to hyphal fusions in fungal colonies

the leopard tree Caesalpinia ferrea (Leguminosae) a native of eastern Brazil-some of the leader branches connect to and fuse with neighbouring branches of the same tree. The bridge initials project out as pegs or protuberances and apparently extend in a coordinated manner, connecting branches up to 4 ft apart. The fusion of two branches of the same tree implies intra-plant communication involving signaling factor(s). The bridges resemble fusions between hyphae in a fungal colony. Whereas hyphal fusions are common and the process is apparently completed in <1 h, branch fusions in C. ferrea tree are limited and a slow process, apparently requiring several months to years to complete. Branch fusions in C. ferrea are in accord with Claus Mattheck's analysis that tree branches actually seek contact rather than avoid contacts.

Role of histidine residues in EcoP15I DNA methyltransferase activity as probed by chemical modification and site-directed mutagenesis

Towards understanding the catalytic mechanism of M.EcoP15I [EcoP15I MTase (DNA methyltransferase); an adenine methyltransferase], we investigated the role of histidine residues in catalysis. M.EcoP15I, when incubated with DEPC (diethyl pyrocarbonate), a histidine-specific reagent, shows a time- and concentration-dependent inactivation of methylation of DNA containing its recognition sequence of 5'-CAGCAG-3'. The loss of enzyme activity was accompanied by an increase in absorbance at 240 nm. A difference spectrum of modified versus native enzyme shows the formation of N-carbethoxyhistidine that is diminished by hydroxylamine. This, along with other experiments, strongly suggests that the inactivation of the enzyme by DEPC was specific for histidine residues. Substrate protection experiments show that pre-incubating the methylase with DNA was able to protect the enzyme from DEPC inactivation. Site-directed mutagenesis experiments in which the 15 histidine residues in the enzyme were replaced individually with alanine corroborated the chemical modification studies and established the importance of His-335 in the methylase activity. No gross structural differences were detected between the native and H335A mutant MTases, as evident from CD spectra, native PAGE pattern or on gel filtration chromatography. Replacement of histidine with alanine residue at position 335 results in a mutant enzyme that is catalytically inactive and binds to DNA more tightly than the wild-type enzyme. Thus we have shown in the present study, through a combination of chemical modification and site-directed mutagenesis experiments, that His-335 plays an essential role in DNA methylation catalysed by M.EcoP15I.

Transcription of Human Resistin Gene Involves an Interaction of Sp1 with Peroxisome Proliferator-Activating Receptor Gamma (PPAR gamma)

Background: Resistin is a cysteine rich protein, mainly expressed and secreted by circulating human mononuclear cells. While several factors responsible for transcription of mouse resistin gene have been identified, not much is known about the factors responsible for the differential expression of human resistin.Methodology/Principal Finding: We show that the minimal promoter of human resistin lies within similar to 80 bp sequence upstream of the transcriptional start site (-240) whereas binding sites for cRel, CCAAT enhancer binding protein alpha (C/EBP-alpha), activating transcription factor 2 (ATF-2) and activator protein 1 (AP-1) transcription factors, important for induced expression, are present within sequences up to -619. Specificity Protein 1(Sp1) binding site (-276 to -295) is also present and an interaction of Sp1 with peroxisome proliferator activating receptor gamma (PPAR gamma) is necessary for constitutive expression in U937 cells. Indeed co-immunoprecipitation assay demonstrated a direct physical interaction of Sp1 with PPAR gamma in whole cell extracts of U937 cells. Phorbol myristate acetate (PMA) upregulated the expression of resistin mRNA in U937 cells by increasing the recruitment of Sp1, ATF-2 and PPAR gamma on the resistin gene promoter. Furthermore, PMA stimulation of U937 cells resulted in the disruption of Sp1 and PPAR gamma interaction. Chromatin immunoprecipitation (ChIP) assay confirmed the recruitment of transcription factors phospho ATF-2, Sp1, Sp3, PPAR gamma, chromatin modifier histone deacetylase 1 (HDAC1) and the acetylated form of histone H3 but not cRel, C/EBP-alpha and phospho c-Jun during resistingene transcription.Conclusion: Our findings suggest a complex interplay of Sp1 and PPAR gamma along with other transcription factors that drives the expression of resistin in human monocytic U937 cells.