Protein Hydration and Water Structure: X-ray Analysis of a Closely Packed Protein Crystal with Very Low Solvent Content

Low-humidity monoclinic lysozyme, resulting from a water-mediated transformation, has one of the lowest solvent contents (22% by volume) observed in a protein crystal. Its structure has been solved by the molecular replacement method and refined to an R value of 0.175 for 7684 observed reflections in the 10–1.75 Å resolution shell. 90% of the solvent in the well ordered crystals could be located. Favourable sites of hydration on the protein surface include side chains with multiple hydrogen-bonding centres, and regions between short hydrophilic side chains and the main-chain CO or NH groups of the same or nearby residues. Major secondary structural features are not disrupted by hydration. However, the free CO groups at the C terminii and, to a lesser extent, the NH groups at the N terminii of helices provide favourable sites for water interactions, as do reverse turns and regions which connect β-structure and helices. The hydration shell consists of discontinuous networks of water molecules, the maximum number of molecules in a network being ten. The substrate-binding cleft is heavily hydrated, as is the main loop region which is stabilized by water interactions. The protein molecules are close packed in the crystals with a molecular coordination number of 14. Arginyl residues are extensively involved in intermolecular hydrogen bonds and water bridges. The water molecules in the crystal are organized into discrete clusters. A distinctive feature of the clusters is the frequent occurrence of three-membered rings. The protein molecules undergo substantial rearrangement during the transformation from the native to the low-humidity form. The main-chain conformations in the two forms are nearly the same, but differences exist in the side-chain conformation. The differences are particularly pronounced in relation to Trp 62 and Trp 63. The shift in Trp 62 is especially interesting as it is also known to move during inhibitor binding.

Effect of zinc content on the processing map for hot working of α brass

The hot deformation behavior of α brass with varying zinc contents in the range 3%–30% was characterized using hot compression testing in the temperature range 600–900 °C and strain rate range 0.001–100 s−1. On the basis of the flow stress data, processing maps showing the variation of the efficiency of power dissipation (given by Image where m is the strain rate sensitivity) with temperature and strain rate were obtained. α brass exhibits a domain of dynamic recrystallization (DRX) at temperatures greater than 0.85Tm and at strain rates lower than 1 s−1. The maximum efficiency of power dissipation increases with increasing zinc content and is in the range 33%–53%. The DRX domain shifts to lower strain rates for higher zinc contents and the strain rate for peak efficiency is in the range 0.0001–0.05 s−1. The results indicate that the DRX in α brass is controlled by the rate of interface formation (nucleation) which depends on the diffusion-controlled process of thermal recovery by climb.

A Geostatistical Study pf Alumina-Content in Ores of An Indian Iron-Ore Deposit

Mining and blending operations in the high grade iron ore deposit under study are performed to optimize recovery with minimal alumina content while maintaining required levels of other chemical component and a proper mix of ore types. In the present work the regionalisation of alumina in the ores has been studied independently and its effects on global and local recoverable tonnage as well as on alternatives of mining operations have been evaluated. The global tonnage recovery curves for blocks (20m x 20m x 12m) obtained by simulation closely approximated the curves obtained theoretically using a change of support under the discretised gaussian model. Variations in block size up to 80m x 20m x 12m did not affect the recovery as the horizontal dimensions of the blocks are small in relation to the range of the variogram. A comparison of the local tonnage recovery curves obtained through multiple conditional simulations made with that obtained by the method of uniform conditioning of block grades on an estimate of panel 100m x 100m x 12m panel grade reveals comparable results only in panels which have been well conditioned and possesing an ensemble simulation mean close to the ordinary kriged value for the panel. Study of simple alternative sequence of mining on the conditionally simulated deposit shows that concentration of mining operations simultaneously on a single bench enhances the fluctuation in alumina values of ore mined.

Metal-Insulator Transitions in Anion-Excess LaMnO3+? Controlled by the Mn4+ Content

LaMnO3+? samples with Mn4+ content up to 50% have been prepared by different methods. The structure of LaMnO3+? changes from orthorhombic to cubic (via rhombohedral) with increase in the Mn4+ content. LaMnO3+? samples containing greater than 20% Mn4+ are ferromagnetic and show resistivity maxima at a temperature Tt which is close to the ferromagnetic Curie temperature. The resistivity maximum is due to the occurrence of a metal-insulator transition. In samples heated to the same temperature, the value of Tt increases with % Mn4+. For a given sample, Tt increases with the temperature of heat treatment due to the increase in particle size. The onset of ferromagnetism in LaMnO3+? accompanied by an insulator-metal transition is similar to that found in La1-xCaxMnO3 and La1-xSrxCoO3.

Characterization of lysozyme crystals with unusually low solvent content

Studies on the low-humidity (88%) forms of tetragonal and monoclinic lysozyme, resulting from water-mediated transformations, have provided a wealth of information on the variability in protein hydration, its structural consequences and the water structure associated with proteins, in addition to facilitating the delineation of the rigid and the flexible regions in the protein molecule and the invariant features in its hydration shell. Surprisingly, monoclinic lysozyme continues to diffract even when the environmental humidity is drastically reduced, thus permitting the structural study of the enzyme at different levels of hydration. As part of a study in this direction, three very low humidity forms, two of them occuring at a nominal relative humidity of 38% and the other at 5% relative humidity, have been characterized. These have unprecedented low solvent contents of 16.9, 17.6 and 9.4%, respectively, as determined by the Matthews method.

Optimization of Lime Content for Fly Ash

Abundant quantities of fly ash have been produced by thermal power plants situated ail over the world. Many applications of fly ash depend upon its pozzolanic reactivity. This reactivity depends upon many factors, including lime content. Many fly ashes show marked improvement with the addition of lime. However, for every fly ash, there is an optimum lime content for its maximum reactivity. There is no well-established simple test to determine the optimum lime content. In this paper an attempt is made to use a simple physical and physico chemical test to determine the optimum lime content. The principle behind the use of a pH test, liquid limit test, and free swell index test to determine the optimum lime content has been explained. All the methods predict nearly the same optimum lime content and correlate well with that determined by the strength test.

Giant magnetoresistance in bulk samples of LaMnO3 with varying Mn4+ content

Magnetoresistance (MR) in bulk samples of LaMnO3 has been investigated by varying the Mn4+ content from 10 to 33 per cent by chemical means, without aliovalent doping. With the increase in Mn4+ content, the structure of LaMnO3 changes first from orthorhombic to rhombohedral and then to cubic and the material becomes increasingly ferromagnetic, exhibiting a resistivity maximum akin to an insulator-metal transition at T-Peak, just below the ferromagnetic T-c. The magnitude of MR is highest in the cubic sample (with 33% Mn4+) around the T-Peak, and negligible in the non-magnetic orthorhombic sample (12% Mn4+).

Variations in the Levels of Aminoacylation and Modified Nucleotide Content Between Total Transfer-RNAS From Chloroplasts and Etioplasts in Cucumber Cotyledons

Total tRNAs isolated from chloroplasts and etioplasts of cucumber cotyledons were compared with respect to amino acid acceptance, isoacceptor distribution and extent of modification. Aminoacylation of the tRNAs with nine different amino acids studied indicated that the relative acceptor activities of chloroplast total tRNAs for four amino acids are significantly higher than etioplast total tRNAs. Two dimensional polyacrylamide gel electrophoresis (2D-PAGE) of chloroplast total tRNAs separated at least 32 spots, while approximately 41 spots were resolved from etioplast total tRNAs. Comparison of the reversed-phase chromatography (RPC-5) profiles of chloroplast and etioplast leucyl-, lysyl-, phenylalanyl-, and valyl-tRNA species showed no qualitative differences in the elution profiles. However, leucyl-, lysyl- and valyl-tRNA species showed quantitative differences in the relative amounts of the isoaccepting species present in chloroplasts and etioplasts. The analysis of modified nucleotides of total tRNAs from the two plastid types indicated that total tRNA from etioplasts was undermodified with respect to ribothymidine, isopentenyladenosine/hydroxy-isopentenyladenosine, 1-methylguanosine and 2-o-methylguanosine. This indicates that illumination may cause de novo synthesis of chloroplast tRNA-modifying enzymes encoded for by nuclear genes leading to the formation of highly modified tRNAs in chloroplasts. Based on these results, we speculate that the observed decrease in levels of aminoacylation, variations in the relative amounts of certain isoacceptors, and differences in the electrophoretic mobilities of some extra tRNA spots in the etioplast total tRNAs as compared to chloroplast total tRNAs could be due to some partially undermodified etioplast tRNAs. Taken together, the data suggested that the light-induced transformation of etioplasts into chloroplasts is accompanied by increases in the relative levels of some functional chloroplast tRNAs by post transcriptional nucleotide modifications.

Structure and activity cyclase activated of OK-GC: A kidney receptor-guanylate cyclase activated by guanylin peptides

Uroguanylin, guanylin, and lymphoguanylin are small peptides that activate renal and intestinal receptor guanylate cyclases (GC). They are structurally similar to bacterial heat-stable enterotoxins (ST) that cause secretory diarrhea. Uroguanylin, guanylin, and ST elicit natriuresis, kaliuresis, and diuresis by direct actions on kidney GC receptors. A 3,762-bp cDNA characterizing a uroguanylin/guanylin/ST receptor was isolated from opossum kidney (OK) cell RNA/cDNA. This kidney cDNA (OK-GC) encodes a mature protein containing 1,049 residues sharing 72.4�75.8% identity with rat, human, and porcine forms of intestinal GC-C receptors. COS or HEK-293 cells expressing OK-GC receptor protein were activated by uroguanylin, guanylin, or ST13 peptides. The 3.8-kb OK-GC mRNA transcript is most abundant in the kidney cortex and intestinal mucosa, with lower mRNA levels observed in urinary bladder, adrenal gland, and myocardium and with no detectable transcripts in skin or stomach mucosa. We propose that OK-GC receptor GC participates in a renal mechanism of action for uroguanylin and/or guanylin in the physiological regulation of urinary sodium, potassium, and water excretion. This renal tubular receptor GC may be a target for circulating uroguanylin in an endocrine link between the intestine and kidney and/or participate in an intrarenal paracrine mechanism for regulation of kidney function via the intracellular second messenger, cGMP.

Electrical transport and magnetic properties of La0.5Ca0.5MnO3-y with varying oxygen content

We report the tuning of oxygen content of La0.5Ca0.5MnO3-y and its effect on electrical transport and magnetic properties. A small reduction of oxygen content leads to a decrease in sample resistivity, which is more dramatic at low temperatures. No significant change is seen to occur in the magnetic properties for this case. Further reduction in the oxygen content increases the resistivity remarkably, as compared to the as-prepared sample. The amplitude of the ferromagnetic (FM) transition at 225 K decreases, and the antiferromagnetic (AFM) transition at 130 K disappears. For samples with y=0.17, insulator-metal transition and paramagnetic-ferromagnetic transition occur around 167 K. The results are explained in terms of the effect of oxygen vacancies on the coexistence of the metallic FM phase and the insulating charge ordered AFM phase.

Expression of the receptor guanylyl cyclase C and its ligands in reproductive tissues of the rat: A potential role for a novel signaling pathway in the epididymis

Guanylyl cyclase C (GC-C) is a membrane-associated form of guanylyl cyclase and serves as the receptor for the heat-stable enterotoxin (ST) peptide and endogenous ligands guanylin, uroguanylin, and lymphoguanylin. The major site of expression of GC-C is the intestinal epithelial cell, although GC-C is also expressed in extraintestinal tissue such as the kidney, airway epithelium, perinatal liver, stomach, brain, and adrenal glands. Binding of ligands to GC-C leads to accumulation of intracellular cGMP, the activation of protein kinases G and A, and phosphorylation of the cystic fibrosis transmembrane conductance regulator (CFTR), a chloride channel that regulates salt and water secretion. We examined the expression of GC-C and its ligands in various tissues of the reproductive tract of the rat. Using reverse transcriptase and the polymerase chain reaction, we demonstrated the presence of GC-C, uroguanylin, and guanylin mRNA in both male and female reproductive organs. Western blot analysis using a monoclonal antibody to GC-C revealed the presence of differentially glycosylated forms of GC-C in the caput and cauda epididymis. Exogenous addition of uroguanylin to minced epididymal tissue resulted in cGMP accumulation, suggesting an autocrine or endocrine activation of GC-C in this tissue. Immunohistochemical analyses demonstrated expression of GC-C in the tubular epithelial cells of both the caput epididymis and cauda epididymis. Our results suggest that the GC-C signaling pathway could converge on CFTR in the epididymis and perhaps control fluid and ion balance for optimal sperm maturation and storage in this tissue.

Insulator-metal transition and magnetoresistance of La0.5Ca0.5MnOy induced by tuning the oxygen content

The oxygen content of La0.5Ca0.5MnOy was tuned by annealing the samples at high temperatures in flowing nitrogen with graphite powder nearby. The reduction of oxygen content has dramatic effect on the electrical transport and magnetic properties. The samples with y=2.983, 2.83, and 2.803 show an insulator-metal transition, and an unusual temperature and magnetic-field dependence of the magnetoresistance. The paramagnetic-ferromagnetic transition also shifts to lower temperatures and the antiferromagnetic transition at lower temperature is suppressed. The results are discussed in terms of the effect of oxygen vacancies on the various properties of La0.5Ca0.5MnOy. (C) 2002 American Institute of Physics.

Influence of martensite content and morphology on the toughness and fatigue behavior of high-martensite dual-phase steels

A series of high-martensite dual-phase (HMDP) steels exhibiting a 0.3 to 0.8 volume fraction of martensite (V m ), produced by intermediate quenching (IQ) of a vanadium and boron-containing microalloyed steel, have been studied for toughness and fatigue behavior to supplement the contents of a recent report by the present authors on the unusual tensile behavior of these steels. The studies included assessment of the quasi-static and dynamic fracture toughness and fatigue-crack growth (FCG) behavior of the developed steels. The experimental results show that the quasi-static fracturetoughness (K ICV ) increases with increasing V m in the range between V m =0.3 and 0.6 and then decreases, whereas the dynamic fracture-toughness parameters (K ID , K D , and J ID ) exhibit a significant increase in their magnitudes for steels containing 0.45 to 0.60 V m before achieving a saturation plateau. Both the quasi-static and dynamic fracture-toughness values exhibit the best range of toughnesses for specimens containing approximately equal amounts of precipitate-free ferrite and martensite in a refined microstructural state. The magnitudes of the fatigue threshold in HMDP steels, for V m between 0.55 and 0.60, appear to be superior to those of structural steels of a similar strength level. The Paris-law exponents (m) for the developed HMDP steels increase with increasing V m , with an attendant decrease in the pre-exponential factor (C).

Water content - void ratio swell-shrink paths of compacted expansive soils

This paper addresses the behaviour of compacted expansive soils under swell-shrink cycles. Laboratory cyclic swell-shrink tests were conducted on compacted specimens of two expansive soils at surcharge pressures of 6.25, 50.00, and 100.00 kPa. The void ratio and water content of the specimens at several intermediate stages during swelling until the end of swelling and during shrinkage until the end of shrinkage were determined to trace the water content versus void ratio paths with an increasing number of swell-shrink cycles. The test results showed that the swell-shrink path was reversible once the soil reached an equilibrium stage where the vertical deformations during swelling and shrinkage were the same. This usually occurred after about four swell-shrink cycles. The swelling and shrinkage path of each specimen subjected to full swelling - full shrinkage cycles showed an S-shaped curve (two curvilinear portions and a linear portion). However, the swelling and shrinkage path occurred as a part of the S-shaped curve, when the specimen was subjected to full swelling - partial shrinkage cycles. More than 80% of the total volumetric change and more than 50% of the total vertical deformation occurred in the central linear portion of the S-shaped curve. The volumetric change was essentially parallel to the saturation line within a degree of saturation range of 50-80% for the equilibrium cycle. The primary value of the swell-shrink path is to provide information regarding the void ratio change that would occur for a given change in water content for any possible swell-shrink pattern. It is suggested that these swell-shrink paths can be established with a limited number of tests in the laboratory.

Role of clay content and moisture on characteristics of cement stabilised rammed earth

Rammed earth is an energy efficient and low carbon emission alternative for load bearing walls. This paper attempts to examine the influence of clay content and moisture content on the compressive strength of cement stabilised rammed earth (CSRE) through experimental investigations. Compressive strength of CSRE prisms was monitored both in dry and wet (saturated) conditions. Major conclusions of the study are:(a) Optimum clay content for maximum compressive strength is about 16%, (b) the strength of CSRE is sensitive to the moisture content at the time of testing, (c) Strength in saturated condition is less than half of the dry strength and (d) Water absorption (saturated water content) increases as the clay content of the soil mix increases and it is in the range of 12 to 16% for the CRSE prisms with 8% cement.