Biochemistry of grape berries: post-genomics approaches to uncover the effects of water deficits on ripening

Dissertation presented to obtain the Ph.D degree in Biochemistry, Plant Physiology

MicroRNAs as modulators of water deficit responses in Medicago truncatula

Dissertação para a obtenção do grau de doutor em Biologia pelo Instituto de Tecnologia Química e Biológica. Universidade Nova de Lisboa

Biotransformations of water insoluble substrates in aqueous, two-phase and encapsulated systems /

The biotransformation of water insoluble substrates by mammalian and bacterial cells has been problematic, since these whole cell reactions are primarily performed in an aqueous environment The implementation of a twophase or encapsulated system has the advantages of providing a low water system along with the physiological environment the cells require to sustain themselves. Encapsulation of mammalian cells by formation of polyamide capsules via interfacial polymerization illustrated that the cells could not survive this type of encapsulation process. Biotransformation of the steroid spironolactone [3] by human kidney carcinoma cells was performed in a substrate-encapsulated system, yielding canrenone [4] in 70% yield. Encapsulation of nitrile-metabolizing Rhodococcus rhodochrous cells using a polyamide membrane yielded leaky capsules, but biotransformation of 2-(4- chlorophenyl)-3-methylbutyronitrile (CPIN) [6] in a free cell system yielded CPIN amide [7] in 40% yield and 94% ee. A two-phase biotransformation of CPIN consisting of a 5:1 ratio of tris buffer, pH 7.2 to octane respectively, gave CPIN acid [8] in 30% yield and 97% ee. It was concluded that Rhodococcus rhodochrous ATCC 17895 contained a nonselective nitrile hydratase and a highly selective amidase enzyme.

Rotation-electronic interaction in the 3p-complex Rydberg state of water

An energy theory is formulated for the rotational energy levels in a p-complex Rydberg state of an asymmetric top molecule of symmetry C2v. The effective Hamiltonian used consists of the usual rigid rotor Hamiltonian augmented with terms representing electronic spin and orbital angular momentum effects. Criteria for assigning symmetry species to the rotational energy levels, following Houganfs scheme that uses the full molecular group,are established and given in the form of a table. This is particularly suitable when eigenvectors are calculated on a digital computer. Also, an intensity theory for transitions to the Rydberg p-complex singlet states is presented and selection rules in terms of symmetry species of energy states are established. Finally, applications to HpO and DpO are given.

The influence of water on the glucose affinity of hexokinase

In the present thesis, the role of hydration during the glucose induced conformational change of hexokinase is investigated. This is accomplished by applying the osmotic stress technique. The osmotic stress technique is founded on varying of the activity of water in a system in order to determine ifs effects. This is accomplished by adding inert solute molecules that are excluded from the system under study. The solute molecules used within the present investigation are Polyethylene glycols (PEGs). PEGs aid in the removal of water from hexokinase by exerting osmotic pressure. The osmotic pressures of the PEG solutions are also measured with both vapour pressure osmometry and secondary osmometry with phospholipids. An interesting discovery is made in that the osmotic pressures of PEG and co-solute solutions are non-additive. This indicates that PEG concentrates co-solutes in solution by making a certain proportion of the water inaccessible. Glucose binding was measured fluorometrically and the glucose equilibrium dissociation constant (GEDC) of hexokinase is measured in solutions containing the different MW PEGs. Changes in the sensitivity of the glucose affinity with osmotic pressure allows the calculation of the change in the numbers of polymer-inaccessible water molecules upon the binding of glucose to hexokinase ~Nw. It was determined the ~Nw decreases with increases in osmotic pressure in the presence of all MW PEGs. ~Nw decreases from values between 45-290 water molecules at low pressure to approximately 15 at high pressure. There is also a molecular weight dependence observed. There are large decreases in ~Nw with osmotic pressure in the presence of PEGs above MW 1000. However, below MW 1500 changes in ~Nw with osmotic pressure are relatively small. These findings are interpreted with respect to two possible mechanisms involving changes in the conformation of hexokinase u~der osmotic pressure and the access of the PEG molecules to water surrounding hexokinase.

The role of water in determining structure and function of macromolecules and macromolecular assemblies /

The interaction of biological molecules with water is an important determinant of structural properties both in molecular assemblies, and in conformation of individual macromolecules. By observing the effects of manipulating the activity of water (which can be accomplished by limiting its concentration or by adding additional solutes, "osmotic stress"), one can learn something about intrinsic physical properties of biological molecules as well as measure an energetic contribution of closely associated water molecules to overall equilibria in biological reactions. Here two such studies are reported. The first of these examines several species of lysolipid which, while present in relatively low concentrations in biomembranes, have been shown to affect many cellular processes involving membrane-protein or membrane-membrane interactions. Monolayer elastic constants were determined by combining X-ray diffraction and the osmotic stress technique. Spontaneous radii of curvature of lysophosphatidylcholines were determined to be positive and in the range +30A to +70A, while lysophosphatidylethanolamines proved to be essentially flat. Neither lysolipid significantly affected the bending modulus of the monolayer in which it was incorporated. The second study examines the role of water in theprocess of polymerization of actin into filaments. Water activity was manipulated by adding osmolytes and the effect on the equilibrium dissociation constant (measured as the criticalmonomer concentration) was determined. As water activity was decreased, the critical concentration was reduced for Ca-actin but not for Mg-actin, suggesting that 10-12 fewer water molecules are associated with Ca-actin in the polymerized state. Thisunexpectedly small amount of water is discussed in the context of the common structural motif of a nucleotide binding cleft.

List tracing Details of Water Race to go through the Lands of Phelps, Clendinnan, Sanderson, Butler, and Chase

List (4 pages of printed material) tracing the details of the water race to go through the lands of Phelps, Clendinnan, Sanderson, Butler and Chase. This includes a copy of the following: the authority dated Dec. 22, 1829; a letter of encouragement dated Oct. 24, 1833; a transfer from Dittrick, Adams and others to the Welland Canal Company dated Dec. 6, 1834; a lease from the Welland Canal Co. to Adams and others dated Nov. 28, 1834 and the terms of the indenture dated May 24, 1847.

Flyer, Rules of the Board of Water Commissioners

Flyer containing the standing Rules of the Board of Water Commissioners for the Town of St. Catharines (2 copies), Nov. 27, 1875.

Power of water discharged over breach of weir

Power of water discharged over breach of weir (1 page, handwritten), n.d.

Investigations on Structural response of Water Backed Perforated plates

The study envisaged herein contains the numerical investigations on Perforated Plate (PP) as well as numerical and experimental investigations on Perforated Plate with Lining (PPL) which has a variety of applications in underwater engineering especially related to defence applications. Finite element method has been adopted as the tool for analysis of PP and PPL. The commercial software ANSYS has been used for static and free vibration response evaluation, whereas ANSYS LS-DYNA has been used for shock analysis. SHELL63, SHELL93, SOLID45, SOLSH190, BEAM188 and FLUID30 finite elements available in the ANSYS library as well as SHELL193 and SOLID194 available in the ANSYS LS-DYNA library have been made use of. Unit cell of the PP and PPL which is a miniature of the original plate with 16 perforations have been used. Based upon the convergence characteristics, the utility of SHELL63 element for the analysis of PP and PPL, and the required mesh density are brought out. The effect of perforation, geometry and orientation of perforation, boundary conditions and lining plate are investigated for various configurations. Stress concentration and deflection factor are also studied. Based on these investigations, stadium geometry perforation with horizontal orientation is recommended for further analysis.Linear and nonlinear static analysis of PP and PPL subjected to unit normal pressure has been carried out besides the free vibration analysis. Shock analysis has also been carried out on these structural components. The analytical model measures 0.9m x 0.9m with stiffener of 0.3m interval. The influence of finite element, boundary conditions, and lining plate on linear static response has been estimated and presented. Comparison of behavior of PP and PPL in the nonlinear strain regime has been made using geometric nonlinear analysis. Free vibration analysis of the PP and PPL has been carried out ‘in vacuum’ condition and in water backed condition, and the influence of water backed condition and effect of perforation on natural frequency have been investigated.Based upon the studies on the vibration characteristics of NPP, PP and PPL in water backed condition and ‘in vacuum’ condition, the reduction in the natural frequency of the plate in immersed condition has been rightly brought out. The necessity to introduce the effect of water medium in the analysis of water backed underwater structure has been highlighted.Shock analysis of PP and PPL for three explosives viz., PEK, TNT and C4 has been carried out and deflection and stresses on plate as well as free field pressure have been estimated using ANSYS LS-DYNA. The effect of perforations and the effect of lining plate have been predicted. Experimental investigations of the measurement of free field pressure using PPL have been conducted in a shock tank. Free field pressure has been measured and has been validated with finite element analysis results. Besides, an experiment has been carried out on PPL, for the comparison of the static deflection predicted by finite element analysis.The distribution of the free field pressure and the estimation of differential pressure from experimentation and the provision for treating the differential pressure as the resistance, as a part of the design load for PPL, has been brought out.

Assessment of Water Quality along the Coastal Areas of Kerala: A Study of the Tsunami Impact on Groundwater

Everywhere, on the coastal belt it is proved without doubt that the pristine ground water quality was severely deteriorated after the 26 December 2004 Indian Ocean Tsunami. But how far is more relevant, as it is decided by the so-called pre-tsunamic situation of the region. In water quality studies it is this reference finger print which earmarks regional ground water chemistry based on which the monthly variability could rationally be interpreted. This Ph D thesis comprises the testing and evaluation of the facts: whether there is any significant difference in the water quality parameters under study between stations and between months in Tsunami Affected Dug Wells (TADW). Whether the selected water quality parameters vary significantly from BIS and WHO standards. Whether the water quality index (WQI) differ significantly between Tsunami Affected Dug Wells (TADW) and Bore Wells (BW). Whether there is any significant difference in the water quality parameters during December 2005 and December 2008. Is there any significant change in the Water Quality Parameters before 2001 and after tsunami (2005) in TADW.

Retting Of Coconut Husk - A Unique Case Of Water Pollution On The South West Coast Of India

The extensive backwaters of Kerala are the sites for a flourishing cottage industry - the coir industry. This enterprise almost exclusively located along the 590 km coastal belt of Kerala, provides direct employment to over half a million people in the state and produces nearly 90% of the total coir goods in the world. The shallow bays and lagoons of the 30 backwater systems of the state are traditional areas for the retting of coconut husk for the production of the coir fibre. The paper examines the environmental status of the retting grounds in Kerala, in relation to the biotic communities. The study revealed that retting activity has caused large scale organic pollution along with the mass destruction of the flora and fauna, converting sizeable sections of the backwaters into virtual cesspools of foul smelling stagnant waters. High values of hydrogen sulphide, ammonia, BOD5 associated with anoxic conditions and low community diversity of plankton, benthic fauna, fish, shell fish, wood boring and fouling organisms were the outstanding feature of the retting zones.

Varying the dietary supply of C and N to manipulate the manure composition of water buffalo heifers in Oman

Optimizing the composition of manure has the potential to reduce nutrient losses to the environment and to increase crop yields. In this study the effect of dietary ratios of carbon (C) to nitrogen (N) and neutral detergent fibre (NDF) to soluble carbohydrates (SC) on faeces composition of water buffalo heifers was assessed. Two digestibility trials were conducted with 12 animals each, fed one control and four test diets composed to achieve (1) high C/N and high NDF/SC ratios (HH), (2) low C/N and low NDF/SC ratios (LL), (3) high C/N and low NDF/SC ratios (HL) and (4) low C/N and high NDF/SC (LH) ratios. Faecal C/N ratios were generally lower than dietary C/N ratios, but the reduction was especially large for high C/N ratio diets (HH=55 %, HL=51 %). Faecal N concentration was positively correlated (r^2 = 0.6; P < 0.001) with N intake, but the increase in faecal N was more pronounced for diets that supplied low amounts of N. Faecal NDF concentration was positively related to NDF intake (r^2 = 0.42; P < 0.001), as well as the faecal C/N ratio (r^2 = 0.3; P < 0.001). Results demonstrate that C/N ratio and NDF concentration of buffalo manure were affected by diet composition. Diets with high C/N ratio and low NDF/SC ratio are preferable with regard to manure quality, but may not satisfy the nutritional requirements of producing animals, since N concentration in these diets was low and fibre concentration simultaneously high.