Structural and functional studies of Bacillus stearothermophilus serine hydroxymethyltransferase: the role of Asn(341), Tyr(60) and Phe(351) in tetrahydrofolate binding

SHMT (serine hydoxymethyltransferase), a type I pyridoxal 5'-phosphate-dependent enzyme, catalyses the conversion of L-serine and THF (tetrahydrofolate) into glycine and 5,10-methylene THE SHMT also catalyses several THF-independent side reactions such as cleavage of P-hydroxy amino acids, trans-amination, racemization and decarboxylation. In the present study, the residues Asn(341), Tyr(60) and Phe(351), which are likely to influence THF binding, were mutated to alanine, alanine and glycine respectively, to elucidate the role of these residues in THF-dependent and -independent reactions catalysed by SHMT. The N341A and Y60A bsSHMT (Bacillus stearothermophilus SHMT) mutants were inactive for the THF-dependent activity, while the mutations had no effect on THF-independent activity. However, mutation of Phe(351) to glycine did not have any effect oil either of the activities. The crystal structures of the glycine binary complexes of the mutants showed that N341A bsSHMT forms an external aldimine as in bsSHMT, whereas Y60A and F351G bsSHMTs exist as a Mixture of internal/external aldimine and gem-diamine forms. Crystal structures of all of the three Mutants obtained in the presence of L-allo-threonine were similar to the respective glycine binary complexes. The structure of the ternary complex of F351G bsSHMT with glycine and FTHF (5-formyl THF) showed that the monoglutamate side chain of FTHF is ordered in both the subunits of the asymmetric unit, unlike in the wild-type bsSHMT. The present studies demonstrate that the residues Asn(341) and Tyr(60) are pivotal for the binding of THF/FTHF, whereas Phe(351) is responsible for the asymmetric binding of FTHF in the two subunits of the dimer.

Studies on glass transition and starch re-crystallization in wheat bread during staling using electrical impedance spectroscopy

Bread staling is a very complex phenomenon that is not yet completely understood. The present work explains how the electrical impedance spectroscopy technique can be utilized to investigate the effect of staling on the physicochemical properties of wheat bread during storage. An instrument based on electrical impedance spectroscopy technique is developed to study the electrical properties of wheat bread both at its crumb and crust with the help of designed multi-channel ring electrodes. Electrical impedance behavior, mainly capacitance and resistance, of wheat bread at crust and crumb during storage (up to 120 h) is investigated. The variation in capacitance showed the glass transition phenomenon at room temperature in bread crust after 96 h of storage with 18% of moisture in it. The resistance changes at bread crumb showed the starch recrystallization during staling.

NMR (1H, 13C) and MO (ab initio, CNDO/2, EHT) studies on basic and N-protonated hydrazinecarbothioamide

The conformational preferences of hydrazinecarbothioamide (HCTA, H2NNHCSNH2) in its basic and N-protonated (PHCTA, H3NNNHCSNH2) forms have been studied by 1H and 13C NMR spectroscopy and by theoretical LCAO-MO methods (ab initio, CNDO/2 and EHT). The hindered rotation around the C---N bond has been investigated by a total line shape analysis for the thioamide protons and by the three MO methods. Changes in the molecular conformation and electronic structure on protonation are briefly discussed.

x-ray studies on crystalline complexes involving amino-acids and peptides .10. head-to-tail sequences in the crystal-structure of l-lysine acetate

l-Lysine acetate crystallises in the monoclinic space group P21 with a = 5.411 (1), b = 7.562(1), c= l2.635(2) Å and β = 91.7(1). The crystal structure was solved by direct methods and refined to an R value of 0.049 using the full matrix least squares method. The conformation and the aggregation of lysine molecules in the structure are similar to those found in the crystal structure of l-lysine l-aspartate. A conspicuous similarity between the crystal structures of l-arginine acetate and l-lysine acetate is that in both cases the strongly basic side chain, although having the largest pK value, interacts with the weakly acidic acetate group leaving the α-amino and the α-carboxylate groups to take part in head-to-tail sequences. These structures thus indicate that electrostatic effects are strongly modulated by other factors so as to give rise to head-to-tail sequences which have earlier been shown to be an almost universal feature of amino acid aggregation in the solid state.

Fluorescence-polarization studies on binding of 4-methylumbelliferyl beta-D-galactopyranoside to Ricinus communis (castor-bean) agglutinin

The binding of Ricinus communis (castor-bean) agglutinin 1 to saccharides was studied by equilibrium dialysis and fluorescence polarization by using the fluorescently labelled sugar 4-methylumbelliferyl beta-D-galactopyranoside. No appreciable change in ligand fluorescence of 4-methylumbelliferyl beta-D-galactopyranoside was considerably polarized on its binding to the lectin. The association constants obtained by Scatchard analysis of equilibrium-dialysis and fluorescence-polarization data do not differ much from each other, and at 25 degrees C, Ka = 2.4 (+/- 0.2) X 10(4)M-1. These values agree reasonably well with that reported in the literature for Ricinus agglutinin 1. The number of binding sites obtained by the different experimental procedures is 1.94 +/- 0.1 per molecule of 120 000 daltons and is equal to the reported value of 2. The consistency in the values of Ka and number of binding sites indicate the absence of additional subsites on Ricinus agglutinin 1 for its specific sugars. In addition, the excellent agreement between the binding parameters obtained by equilibrium dialysis and fluorescence polarization indicate the potential of ligand-fluorescence-polarization measurements in the investigation of lectin-sugar interactions.

The paribhasas arthavadgrahane nanarthakasya, laksanapratipadoktayoh pratipadoktasyaiva grahanam and ekadesavikrtam ananyavat : Studies on some Metarules in Paninian system

In his grammatical treatise, the Astadhyayi, Panini includes sutras that state guiding rules for the right interpretation and application of his other directly grammatical sutras. These sutras are called paribhasas. In addition to these paribhasas, the various commentaries on Panini frequently invoke supplementary paribhasas which are not stated explicitly in his Astadhyayi. These paribhasas have been a subject of study since early times after Panini and have also occupied modern scholars on Panini s grammar. In regard to most of them, it remains unsettled even today whether they are used in the Astadhyayi, where they apply, what is their role, and whether they are necessary in arriving at the desired grammatical form. Some scholars go even further and argue that none of such paribhasas were intended by Panini. This study aims to settle this question by dealing with three such of these paribhasas individually considering all the information available in the commentaries in their regard and examining the cases in which, according to commentaries, the paribhasas apply. I select the paribhasas arthavadgrahane nanarthakasya, laksanapratipadoktayoh pratipadoktasyaiva grahanam and ekadesavikrtam ananyavat, which are all considered nyayasiddha or lokanyayasiddha; they express logical and obvious principles which are found in daily life. On this basis, Paniniyas explain why Panini did not mention them in the Astadhyayi. I discuss each paribhasa separately and all the issues it involves. I present and explain the cases where the specified paribhasas are invoked in the major commentaries, the Mahabhasya, the Kasika and the Siddhantakaumudi and the arguments found in the commentaries concerning these cases. If available, I supply other solutions to the difficulties for which these paribhasas are invoked. The study aims to make the issue of these paribhasas clearer, which will help us to reach a solution to the key question, that is, whether Panini has presupposed them in his Astadhyayi. My study shows that Panini has presupposed the paribhasa ekadesavikrtam ananyavat (or a similar principle). He also may have used the paribhasa arthavadgrahane nanarthakasya (or a similar principle) as this paribhasa does not lead to undesired results. As for the paribhasa laksanapratipadoktayoh pratipadoktasyaiva grahanam (or a similar principle), the original scope of this paribhasa was clearly extended by later Paniniyas. Moreover, their interpretation of this paribhasa conflicts with Panini s procedure. If Panini has used this paribhasa, he has used it in a very limited way.

Use of S1 nuclease and formamide in combination for the reassociation studies on GC-rich DNA

The optimal parameters in the use of nuclease S1 in DNA reassociation kinetics in the presence of formamide have been determined. The conditions are especially suitable for the study of DNA rich in mole percent GC. A 10-fold dilution of the reassociation samples leading to a decrease in both NaCl and formamide concentrations, consequently resulting in a lowering of Tm by only 1.5°C, and the S1 digestion at temperatures identical to the reassociation assay in order to retain the stability of the duplex, are two important aspects of this system. Under these conditions, the kinetics of reassociation followed the theoretically predicted pattern, while the earlier reported methods have shown lower values.

Studies on the Tracking and Erosion Resistance of RTV Silicone Rubber Nanocomposite

One of the problems associated with outdoor polymeric insulators is tracking and erosion of the weathershed which can directly influence the reliability of the power system. Flame retardants are added to the base material to enhance its tracking and erosion resistance. Hydroxide fillers are regarded as the best flame retardants. This paper deals with studies related to nano - sized magnesium dihydroxide (MDH) and micron-sized Alumina Trihydrate (ATH) fillers as flame retardants in RTV silicone rubber. Tracking and erosion resistance studies were carried out on MDH and ATH silicone rubber composites using an inclined plane tracking and erosion (IPT) resistance tester. The MDH filled (5% by wt) composites performed much better than ATH composites in terms of eroded mass, depth of erosion, width and length of erosion. The eroded mass of MDH composite is 49.8 % that of ATH composite which can be attributed to high surface area and higher thermal stability of MDH nanofillers.

X-ray Studies on Crystalline Complexes Involving Amino Acids and Peptides. XIII. Effect of Chirality on Molecular Aggregation: The Crystal Structures of L-Arginine D-Aspartate and L-Arginine D-Glutamate Trihydrate

The crystal structures of (1) L-arginine D-asparate, C6HIsN40~.C4H6NO4 [triclinic, P1, a=5.239(1), b=9.544(1), c=14.064(2)A, a=85"58(1), /3=88.73 (1), ~/=84.35 (1) °, Z=2] and (2) L-arginine D-glutamate trihydrate, C6H15N40~-.CsHsNO4.3H20 [monoclinic, P2~, a=9.968(2), b=4.652(1), c=19.930 (2) A, fl = 101.20 (1) °, Z = 2] have been determined using direct methods. They have been refined to R =0.042 and 0.048 for 2829 and 2035 unique reflections respectively [I>2cr(I)]. The conformations of the two arginine molecules in the aspartate complex are different from those observed so far in the crystal structures of arginine, its salts and complexes. In both complexes, the molecules are organized into double layers stacked along the longest axis. The core of each double layer consists of two parallel sheets made up of main-chain atoms, each involving both types of molecules. The hydrogen bonds within each sheet and those that interconnect the two sheets give rise to EL-, DD- and DE-type head-to-tail sequences. Adjacent double layers in (1) are held together by side-chain-side-chain interactions whereas those in (2) are interconnected through an extensive network of water molecules which interact with sidechain guanidyl and carboxylate groups. The aggregation pattern observed in the two LD complexes is fundamentally different from that found in the corresponding EL complexes.

Electron and x-ray diffraction studies on Al86Fe14, Al82Fe18 and Al75Fe25 quasicrystals

Electron and x-ray diffraction experiments on the metlt-spun Al100−x Fe x (x=14, 18, 25) alloys are carried out. It is observed that all the melt-spun alloys possessing the quasi-crystalline phases have icosahedral point-group symmetry.

Nonverbal predication in Erzya : Studies on morphosyntactic variation and part of speech distinctions

This dissertation consists of four articles and an introduction. The five parts address the same topic, nonverbal predication in Erzya, from different perspectives. The work is at the same time linguistic typology and Uralic studies. The findings based on a large corpus of empirical Erzya data, which was collected using several different methods and included recordings of the spoken language, made it possible for the present study to apply, then test and finally discuss the previous theories based on cross-linguistic data. Erzya makes use of multiple predication patterns which vary from totally analytic to the morphologically very complex. Nonverbal predicate clause types are classified on the basis of propositional acts in clauses denoting class-membership, identity, property and location. The predicates of these clauses are nouns, adjectives and locational expressions, respectively. The following three predication strategies in Erzya nonverbal predication can be identified: i. the zero-copula construction, ii. the predicative suffix construction and iii. the copula construction. It has been suggested that verbs and nouns cannot be clearly distinguished on morphological grounds when functioning as predicates in Erzya. This study shows that even though predicativity must not be considered a sufficient tool for defining parts of speech in any language, the Erzya lexical classes of adjective, noun and verb can be distinguished from each other also in predicate position. The relative frequency and degree of obligation for using the predicative suffix construction decreases when moving left to right on the scale verb adjective/locative noun ( identificational statement). The predicative suffix is the main pattern in the present tense over the whole domain of nonverbal predication in Standard Erzya, but if it is replaced it is most likely to be with a zero-copula construction in a nominal predication. This study exploits the theory of (a)symmetry for the first time in order to describe verbal vs. nonverbal predication. It is shown that the asymmetry of paradigms and constructions differentiates the lexical classes. Asymmetrical structures are motivated by functional level asymmetry. Variation in predication as such adds to the complexity of the grammar. When symmetric structures are employed, the functional complexity of grammar decreases, even though morphological complexity increases. The genre affects the employment of predication strategies in Erzya. There are differences in the relative frequency of the patterns, and some patterns are totally lacking from some of the data. The clearest difference is that the past tense predicative suffix construction occurs relatively frequently in Standard Erzya, while it occurs infrequently in the other data. Also, the predicative suffixes of the present tense are used more regularly in written Standard Erzya than in any other genre. The genre also affects the incidence of the translative in uľ(ń)ems copula constructions. In translations from Russian to Erzya the translative case is employed relatively frequently in comparison to other data. This study reveals differences between the two Mordvinic languages Erzya and Moksha. The predicative suffixes (bound person markers) of the present tense are used more regularly in Moksha in all kinds of nonverbal predicate clauses compared to Erzya. It should further be observed that identificational statements are encoded with a predicative suffix in Moksha, but seldom in Erzya. Erzya clauses are more frequently encoded using zero-constructions, displaying agreement in number only.

Personality and social psychological studies on computer-mediated communication in school settings

The present study examined how personality and social psychological factors affect third and fourth graders' computer-mediated communication. Personality was analysed in terms of the following strategies: optimism, pessimism and defensive pessimism. Students worked either individually or in dyads which were paired homogeneously or heterogeneously according to the strategies. Moreover, the present study compared horizontal and vertical interaction. The study also examined the role that popularity plays, and students were divided into groups based on their popularity level. The results show that an optimistic strategy is useful. Optimism was found to be related to the active production and processing of ideas. Although previous research has identified drawbacks to pessimism in achievement settings, this study shows that the pessimistic strategy is not as debilitating a strategy as is usually assumed. Pessimistic students were able to process their ideas. However, defensive pessimists were somewhat cautious in introducing or changing ideas. Heterogeneous dyads were not beneficial configurations with respect to producing, introducing, or changing ideas. Moreover, many differences were found to exist between the horizontal and vertical interaction; specifically, the students expressed more opinions and feelings when teachers took no part in the discussions. Strong emotions were observed especially in the horizontal interaction. Further, group working skills were found to be more important for boys than for girls, while rejected students were not at a disadvantage compared to popular ones. Schools can encourage emotional and social learning. The present study shows that students can use computers to express their feelings. In addition, students who are unpopular in non-computer contexts or students who use pessimism can benefit from computers. Participation in computer discussions can give unpopular children a chance to develop confidence when relating to peers.

Studies on vacuum pyrolysis of ammonium perchlorate and ammonium perchlorate/polystyrene propellant in the presence of transition metal oxides

Vacuum pyrolysis of ammonium perchlorate (AP) and ammonium perchlorate/polystyrene (PS) propellant has been studied by differential thermal analysis (DTA) in order to observe the effect of transition metal oxides on sublimation. Sublimation and decomposition being competitive processes, their proportions depend on the pressure of the pyrolysis chamber. The enthalpies for complete decomposition and complete sublimation are available from the literature and by using these data together with DTA area measurements, the extents of sublimation and decomposition have been calculated for AP and the propellant system. The effect of the metal ions on the extent and rate of sublimation depends on their nature. For AP the extent of sublimation increases with a decrease in particle size. For the propellants the powder sublimes more readily than the bulk material, but in the presence of metal ions the bulk material sublimes more readily than the powder. To substantiate this finding, the effect of MnO2 on AP sublimation as a function of particle size was examined, and it was observed that the extent of sublimation decreases as the particle size decreases.

Mechanistic studies on the pyrolysis and combustion of polystyrene/ammonium perchlorate propellants in the presence of transition metal oxides

The effect of transition metal oxides (Fe2O3, MnO2, Ni2O3 and Co2O3) on polystyrene/ammonium perchlorate propellant systems has been examined. The mechanism of action of the oxides in increasing the burning rate was examined by studying the effect of the oxides on the thermal decomposition and combustion of the oxidizer and the propellant. It has been concluded that one of the mechanisms by which the oxides act is by promoting the charge-transfer process, which is indicated by the enhancement of the electron-transfer process in ammonium perchlorate and by the correlation between the redox potential of the metal ions and the corresponding burning rates of the propellant.

NOx Reduction and Desorption Studies Under Electric Discharge Plasma Using a Simulated Gas Mixture: A Case Study on the Effect of Corona Electrodes

In this study, reduction and desorption of oxides of nitrogen (NOx) were conducted using an electrical discharge plasma technique. The study was carried out using a simulated gas mixture to explore the possibility of re-generation of used adsorbents by a nonthermal plasma desorption technique. Three different types of corona electrodes, namely, pipe, helical wire, and straight wire, were used for analyzing their effectiveness in NOx reduction/desorption. The pipe-type corona electrode exhibited a nitric oxide (NO) conversion of 50%, which is 1.5 times that of the straight-wire-type electrode at an energy density of 175J/L. The helical-wire-type corona electrode exhibited a NOx desorption efficiency almost 4 times that of the pipe-type electrode,indicating the possibility that corona-generated species play a crucial role in desorption.