Studies on nucleotidases in plants: Fluorescence and kinetic properties of nucleotide pyrophosphatase from mung bean (Phaseolus aureus) seedlings

Nucleotide pyrophosphatase of mung bean seedlings has earlier been isolated in our laboratory in a dimeric form (Mr 65,000) and has been shown to be converted to a tetramer by AMP and to a monomer by p-hydroxymercuribenzoate. All the molecular forms were enzymatically active with different kinetic properties. By a modified procedure using blue-Sepharose affinity chromatography, we have now obtained a dimeric form of the enzyme which is desensitized to AMP interaction. The molecular weight of the desensitized form of the enzyme was found to be the same as that of the native dimeric enzyme. However, the desensitized enzyme functioned with a linear time course, contrary to the biphasic time course exhibited by the native enzyme. In addition, it was not converted to a tetramer on the addition of AMP, had only one binding site for adenine nucleotides, and p-hydroxy-mercuribenzoate had no effect on the time course of the reaction or on the molecular weight of the enzyme. The temperature optimum of the desensitized enzyme was found to be 67 °C in contrast to the optimum of 49 °C for the native dimer. Fifty percent of the tryptophan residues of the desensitized enzyme were not accessible for quenching by iodide. Fluorescence studies gave Kd values of 0.34, 2.2, and 0.8 mImage for AMP, ADP, and ATP, which were close to the Ki values of 0.12, 2.2, and 0.9 mImage , respectively, for these nucleotides. The binding and inhibition studies with AMP and its analogs showed that the 6-amino group and the 5′-phosphate group were essential for the inhibition of the enzyme activity.

Studies in biogas technology. Part III. Thermal analysis of biogas plants

A thermal model for a conventional biogas plant has been developed in order to understand the heat transfer from the slurry and the gas holder to the surrounding earth and air respectively. The computations have been performed for two conditions : (i) when the slurry is at an ambient temperature of 20°C, and (ii) when it is at 35°C, the optimum temperature for anaerobic fermentation. Under both these conditions, the gas holder is the major “culprit” with regard to heat losses from the biogas plant. The calculations provide an estimate for the heat which has to be supplied by external means to compensate for the net heat losses which occur if the slurry is to be maintained at 35°C. Even if this external supply of heat is realised through (the calorific value of) biogas, there is a net increase in the biogas output, and therefore a net benefit, by operating the plant at 35°C. At this elevated temperature, the cooling effect of adding the influent at ambient temperature is not insignificant. In conclusion, the results of the thermal analysis are used to define a strategy for operating biogas plants at optimum temperatures, or at higher temperatures than the ambient.

Standardization of pollen allergens of Parthenium hysterophoms and selection of an in-house reference extract

A standardized in-house reference extract from the pollen of Parthenium hysterophorus, which is responsible for the high incidence of allergic rhinitis in India, was generated and examined by skin test, radio-allergosorbent test inhibition and isoelectric focusing. Parthenium reference allergen discs and positive reference serum were also generated. These reference reagents could not only be used for the quantitation of Parthenium-specific IgE in the sera of rhinitis patients but also for the evaluation of allergenic activity (relative potency and lot-to-lot variation) of different batches of Parthenium pollen.

Studies on molecular evolution and structural features of double-headed inhibitors of ?-amylase and trypsin in plants

Plant seeds usually have high concentrations of proteinase and amylase inhibitors. These inhibitors exhibit a wide range of specificity, stability and oligomeric structure. In this communication, we report analysis of sequences that show statistically significant similarity to the double-headed alpha-amylase/trypsin inhibitor of ragi (Eleusine coracana). Our aim is to understand their evolutionary and structural features. The 14 sequences of this family that are available in the SWISSPROT database form three evolutionarily distinct branches. The branches relate to enzyme specificities and also probably to the oligomeric state of the proteins and not to the botanical class of the plant from which the enzymes are derived. This suggests that the enzyme specificities of the inhibitors evolved before the divergence of commercially cultivated cereals. The inhibitor sequences have three regions that display periodicity in hydrophobicity. It is likely that this feature reflects extended secondary structure in these segments. One of the most variable regions of the polypeptide corresponds to a loop, which is most probably exposed in the native structure of the inhibitors and is responsible for the inhibitory property.

Immune responses to hemagglutinin-neuraminidase protein of peste des petits ruminants virus expressed in transgenic peanut plants in sheep

Peste des petits ruminants (PPR) is an acute, highly contagious disease of small ruminants caused by a morbillivirus, Peste des petits ruminants virus (PPRV). The disease is prevalent in equatorial Africa, the Middle East, and the Indian subcontinent. A live attenuated vaccine is in use in some of the countries and has been shown to provide protection for at least three years against PPR. However, the live attenuated vaccine is not robust in terms of thermotolerance. As a step towards development of a heat stable subunit vaccine, we have expressed a hemagglutinin-neuraminidase (HN) protein of PPRV in peanut plants (Arachis hypogea) in a biologically active form, possessing neuraminidase activity. Importantly. HN protein expressed in peanut plants retained its immunodominant epitopes in their natural conformation. The immunogenicity of the plant derived HN protein was analyzed in sheep upon oral immunization. Virus neutralizing antibody responses were elicited upon oral immunization of sheep in the absence of any mucosal adjuvant. In addition, anti-PPRV-HN specific cell-mediated immune responses were also detected in mucosally immunized sheep. (C) 2010 Elsevier B.V. All rights reserved.

An ethnobotanical survey of medicinal plants in the Eastern Himalayan zone of Arunachal Pradesh, India

Aim of the study: The medicinal plants are integral source of easily available remedy used in rural healthcare system. This study was conducted among three major ethnic groups namely the Nocte, the Nyishi and the Adi in the Eastern Himalayan region of Arunachal Pradesh to evaluate their comparative knowledge on medicinal plants. Materials and methods: The three remote districts of Arunachal Pradesh namely the Tirap, the Dibang Valley and the Papum Pare were surveyed through interviewing of randomly selected 237 participants using semi-structured questionnaire and regular field visits to selected districts. Results: We recorded the traditional use of 74 medicinal plants species belonging to 41 taxonomic plant families used for treating a total of 25 different diseases/ailments. The informant consensus factor (ICF) values demonstrated that local people tend to agree more with each other in terms of the plants used to treat malaria (0.71), jaundice (0.62), urological problems (0.56), dermatological disorders (0.45), pain (0.30), and respiratory disorder (0.33), and while the general health (0.15) and gastro-intestinal disorders category (0.28) were found low ICF values. Conclusion: Of the total 74 species recorded, the highest number of medicinal plants (36 species) was reported from the Adi of Lower Dibang Valley followed by the Nocte of the Tirap (25 species) and the Nyishi ethnic groups of Papum Pare districts (13 species). In the present study, we found that the men, elder people and illiterate ones had better knowledge on medicinal plants as compared to women, younger and literate people. Findings of this documentation study can be used as an ethnopharmacological basis for selecting plants for future phytochemical and pharmaceutical studies. (C) 2010 Elsevier Ireland Ltd. All rights reserved.

Locating and sizing plants for bottling propane in south India

Shri Shakti LPG Ltd. (SSLPG) imports and markets propane (referred to as liquefied petroleum gas (LPG) in India) in south India. It sells LPG in packed (cylinder) form to domestic customers and commercial establishments through a network of dealers. Dealers replenish their stocks of filled cylinders from bottling plants, which in turn receive LPG in bulk from the cheaper of SSLPG's two import-and-storage facilities that are located on the Indian coast. We implemented integer programming to help SSLPG decide on the locations and long-run sizes of its bottling plants. We estimate that our recommended configuration of bottling plants is about $1 million cheaper annually than the one that SSLPG had initially planned.

An adaptive cascade compensator for a wide class of stable minimum or nonminimum phase plants through time moment estimation

Model Reference Adaptive Control (MRAC) of a wide repertoire of stable Linear Time Invariant (LTI) systems is addressed here. Even an upper bound on the order of the finite-dimensional system is unavailable. Further, the unknown plant is permitted to have both minimum phase and nonminimum phase zeros. Model following with reference to a completely specified reference model excited by a class of piecewise continuous bounded signals is the goal. The problem is approached by taking recourse to the time moments representation of an LTI system. The treatment here is confined to Single-Input Single-Output (SISO) systems. The adaptive controller is built upon an on-line scheme for time moment estimation of a system given no more than its input and output. As a first step, a cascade compensator is devised. The primary contribution lies in developing a unified framework to eventually address with more finesse the problem of adaptive control of a large family of plants allowed to be minimum or nonminimum phase. Thus, the scheme presented in this paper is confined to lay the basis for more refined compensators-cascade, feedback and both-initially for SISO systems and progressively for Multi-Input Multi-Output (MIMO) systems. Simulations are presented.

The selection of a sugar for transport and storage of carbon in plants

Sugars perform two vital functions in plants: as compatible solutes protecting the cell against osmotic stress and as mobile source of immediate and long-term energy requirement for growth and development. The two sugars that occur commonly in nature are sucrose and trehalose. Sucrose comprises one glucose and one fructose molecule; trehalose comprises two glucose molecules. Trehalose occurs in significant amounts in insects and fungi which greatly outnumber the plants. Surprisingly, in plants trehalose has been found in barely detectable amounts, if at all, raising the question `why did nature select sucrose instead of trehalose as the mobile energy source and as storage sugar for the plants'? Modelling revealed that when attached to the ribbon-shaped beta-1,4 glucan a trehalose molecule is shaped like a hook. This suggests that the beta-1,4 glucan chains with attached trehalose will fail to align to form inter-chain hydrogen bonds and coalesce into a cellulose microfibril, as a result of which in trehalose-accumulating plant cells, the cell wall will tend to become leaky. Thus in plants an evolutionary selection was made in favour of sucrose as the mobile energy source. Genetic engineering of plant cells for combating abiotic stresses through microbial trehalose-producing genes is fraught with risk of damage to plant cell walls.

Time dependent deformations in concrete: A multi-scale approach

Estimation of creep and shrinkage are critical in order to compute loss of prestress with time in order to compute leak tightness and assess safety margins available in containment structures of nuclear power plants. Short-term creep and shrinkage experiments have been conducted using in-house test facilities developed specifically for the present research program on 35 and 45 MPa normal concrete and 25 MPa heavy density concrete. The extensive experimental program for creep, has cylinders subject to sustained levels of load typically for several days duration (till negligible strain increase with time is observed in the creep specimen), to provide the total creep strain versus time curves for the two normal density concrete grades and one heavy density concrete grade at different load levels, different ages at loading, and at different relative humidity’s. Shrinkage studies on prism specimen for concrete of the same mix grades are also being studied. In the first instance, creep and shrinkage prediction models reported in the literature has been used to predict the creep and shrinkage levels in subsequent experimental data with acceptable accuracy. While macro-scale short experiments and analytical model development to estimate time dependent deformation under sustained loads over long term, accounting for the composite rheology through the influence of parameters such as the characteristic strength, age of concrete at loading, relative humidity, temperature, mix proportion (cement: fine aggregate: coarse aggregate: water) and volume to surface ratio and the associated uncertainties in these variables form one part of the study, it is widely believed that strength, early age rheology, creep and shrinkage are affected by the material properties at the nano-scale that are not well established. In order to understand and improve cement and concrete properties, investigation of the nanostructure of the composite and how it relates to the local mechanical properties is being undertaken. While results of creep and shrinkage obtained at macro-scale and their predictions through rheological modeling are satisfactory, the nano and micro indenting experimental and analytical studies are presently underway. Computational mechanics based models for creep and shrinkage in concrete must necessarily account for numerous parameters that impact their short and long term response. A Kelvin type model with several elements representing the influence of various factors that impact the behaviour is under development. The immediate short term deformation (elastic response), effects of relative humidity and temperature, volume to surface ratio, water cement ratio and aggregate cement ratio, load levels and age of concrete at loading are parameters accounted for in this model. Inputs to this model, such as the pore structure and mechanical properties at micro/nano scale have been taken from scanning electron microscopy and micro/nano-indenting of the sample specimen.