The Hinge Region of Human Thyroid-Stimulating Hormone (TSH) Receptor Operates as a Tunable Switch between Hormone Binding and Receptor Activation

The mechanism by which the hinge regions of glycoprotein hormone receptors couple hormone binding to activation of downstream effecters is not clearly understood. In the present study, agonistic (311.62) and antagonistic (311.87) monoclonal antibodies (MAbs) directed against the TSH receptor extracellular domain were used to elucidate role of the hinge region in receptor activation. MAb 311.62 which identifies the LRR/Cb-2 junction (aa 265-275), increased the affinity of TSHR for the hormone while concomitantly decreasing its efficacy, whereas MAb 311.87 recognizing LRR 7-9 (aa 201-259) acted as a non-competitive inhibitor of Thyroid stimulating hormone (TSH) binding. Binding of MAbs was sensitive to the conformational changes caused by the activating and inactivating mutations and exhibited differential effects on hormone binding and response of these mutants. By studying the effects of these MAbs on truncation and chimeric mutants of thyroid stimulating hormone receptor (TSHR), this study confirms the tethered inverse agonistic role played by the hinge region and maps the interactions between TSHR hinge region and exoloops responsible for maintenance of the receptor in its basal state. Mechanistic studies on the antibody-receptor interactions suggest that MAb 311.87 is an allosteric insurmountable antagonist and inhibits initiation of the hormone induced conformational changes in the hinge region, whereas MAb 311.62 acts as a partial agonist that recognizes a conformational epitope critical for coupling of hormone binding to receptor activation. The hinge region, probably in close proximity with the alpha-subunit in the hormone-receptor complex, acts as a tunable switch between hormone binding and receptor activation.

The antibodies against the computationally designed mimic of the glycoprotein hormone receptor transmembrane domain provide insights into receptor activation and suppress the constitutively activated receptor Mutants

The exoloops of glycoprotein hormone receptors (GpHRs) transduce the signal generated by the ligand-ectodomain interactions to the transmembrane helices either through direct hormonal contact and/or by modulating the interdomain interactions between the hinge region (HinR) and the transmembrane domain (TMD). The ligand-induced conformational alterations in the HinRs and the interhelical loops of luteinizing hormone receptor/follicle stimulating hormone receptor/thyroid stimulating hormone receptor were mapped using exoloop-specific antibodies generated against a mini-TMD protein designed to mimic the native exoloop conformations that were created by joining the thyroid stimulating hormone receptor exoloops constrained through helical tethers and library-derived linkers. The antibody against the mini-TMD specifically recognized all three GpHRs and inhibited the basal and hormone-stimulated cAMP production without affecting hormone binding. Interestingly, binding of the antibody to all three receptors was abolished by prior incubation of the receptors with the respective hormones, suggesting that the exoloops are buried in the hormone-receptor complexes. The antibody also suppressed the high basal activities of gain-of-function mutations in the HinRs, exoloops, and TMDs such as those involved in precocious puberty and thyroid toxic adenomas. Using the antibody and point/deletion/chimeric receptor mutants, we demonstrate that changes in the HinR-exoloop interactions play an important role in receptor activation. Computational analysis suggests that the mini-TMD antibodies act by conformationally locking the transmembrane helices by means of restraining the exoloops and the juxta-membrane regions. Using GpHRs as a model, we describe a novel computational approach of generating soluble TMD mimics that can be used to explain the role of exoloops during receptor activation and their interplay with TMDs.

Oxidative DNA cleavage, cytotoxicity and antimicrobial studies of L-ornithine copper (II) complexes

New ternary copper (II) complexes, Cu(L-orn)(B)(Cl)](Cl center dot 2H(2)O) (1-2) where L-orn is L-ornithine, B is an N,N-donor heterocyclic base, viz. 2,2'-bipyridine (bpy, 1) and 1,10-phenanthroline (phen, 2), were synthesized and characterized by various spectroscopic techniques. Complex 2 is characterized by the X-ray single crystallographic method. The complex shows a distorted square-pyramidal (4 + 1) CuN3OCl coordination sphere. Binding interactions of the complexes with calf thymus DNA (CT-DNA) were investigated by UV-Vis absorption titration, ethidium bromide displacement assay, viscometric titration experiment and DNA melting studies. Complex 2 shows appreciable chemical nuclease activity in the presence of 3-mercaptopropionic acid (MPA). The complexes were subjected to in vitro cytotoxicity studies against carcinomic human alveolar basal epithelial cells (A-549) and human epithelial (HEp-2) cells. The IC50 values of 1 and 2 are less than that of cisplatin against HEp-2 cell lines. MIC values for 1 against the bacterial strains Streptococcus mutans and Pseudomonas aeruginosa are 0.5 mM. (C) 2012 Elsevier Ltd. All rights reserved.

Evolution of Texture and Microstructure in Commercially Pure Titanium with Change in Strain Path During Rolling

The evolution of microstructure and texture in commercially pure titanium has been studied as a function of strain path during rolling using experimental techniques and viscoplastic self-consistent simulations. Four different strain paths, namely unidirectional rolling, two-step cross rolling, multistep cross rolling, and reverse rolling, have been employed to decipher the effect of strain path change on the evolution of deformation texture and microstructure. The cross-rolled samples show higher hardness with lower microstrain and intragranular misorientation compared to the unidirectional rolled sample as determined from X-ray diffraction and electron backscatter diffraction, respectively. The higher hardness of the cross-rolled samples is attributed to orientation hardening due to the near basal texture. Viscoplastic self-consistent simulations are able to successfully predict the texture evolution of the differently rolled samples. Simulation results indicate the higher contribution of basal slip in the formation of near basal texture and as well as lower intragranular misorientation in the cross-rolled samples.

Performance of established native seedlings in relation to invasive Lantana camara, rainfall and species' habitat preferences in a seasonally dry tropical forest

Native species' response to the presence of invasive species is context specific. This response cannot be studied in isolation from the prevailing environmental stresses in invaded habitats such as seasonal drought. We investigated the combined effects of an invasive shrub Lantana camara L. (lantana), seasonal rainfall and species' microsite preferences on the growth and survival of 1,105 naturally established seedlings of native trees and shrubs in a seasonally dry tropical forest. Individuals were followed from April 2008 to February 2010, and growth and survival measured in relation to lantana density, seasonality of rainfall and species characteristics in a 50-ha permanent forest plot located in Mudumalai, southern India. We used a mixed effects modelling approach to examine seedling growth and generalized linear models to examine seedling survival. The overall relative height growth rate of established seedlings was found to be very low irrespective of the presence or absence of dense lantana. 22-month growth rate of dry forest species was lower under dense lantana while moist forest species were not affected by the presence of lantana thickets. 4-month growth rates of all species increased with increasing inter-census rainfall. Community results may be influenced by responses of the most abundant species, Catunaregam spinosa, whose growth rates were always lower under dense lantana. Overall seedling survival was high, increased with increasing rainfall and was higher for species with dry forest preference than for species with moist forest preference. The high survival rates of naturally established seedlings combined with their basal sprouting ability in this forest could enable the persistence of woody species in the face of invasive species.

Predicting the lithospheric stress field and plate motions by joint modeling of lithosphere and mantle dynamics

The way in which basal tractions, associated with mantle convection, couples with the lithosphere is a fundamental problem in geodynamics. A successful lithosphere-mantle coupling model for the Earth will satisfy observations of plate motions, intraplate stresses, and the plate boundary zone deformation. We solve the depth integrated three-dimensional force balance equations in a global finite element model that takes into account effects of both topography and shallow lithosphere structure as well as tractions originating from deeper mantle convection. The contribution from topography and lithosphere structure is estimated by calculating gravitational potential energy differences. The basal tractions are derived from a fully dynamic flow model with both radial and lateral viscosity variations. We simultaneously fit stresses and plate motions in order to delineate a best-fit lithosphere-mantle coupling model. We use both the World Stress Map and the Global Strain Rate Model to constrain the models. We find that a strongly coupled model with a stiff lithosphere and 3-4 orders of lateral viscosity variations in the lithosphere are best able to match the observational constraints. Our predicted deviatoric stresses, which are dominated by contribution from mantle tractions, range between 20-70 MPa. The best-fitting coupled models predict strain rates that are consistent with observations. That is, the intraplate areas are nearly rigid whereas plate boundaries and some other continental deformation zones display high strain rates. Comparison of mantle tractions and surface velocities indicate that in most areas tractions are driving, although in a few regions, including western North America, tractions are resistive. Citation: Ghosh, A., W. E. Holt, and L. M. Wen (2013), Predicting the lithospheric stress field and plate motions by joint modeling of lithosphere and mantle dynamics.

Microstructure and texture evolution during sub-transus thermomechanical processing of Ti-6Al-4V-0.1B alloy: part I. hot rolling in (alpha plus beta) phase field

In the current study, the evolution of microstructure and texture has been studied for Ti-6Al-4V-0.1B alloy during sub-transus thermomechanical processing. This part of the work deals with the deformation response of the alloy by rolling in the (alpha + beta) phase field. The (alpha + beta) annealing behavior of the rolled specimen is communicated in part II. Rolled microstructures of the alloys exhibit either kinked or straight alpha colonies depending on their orientations with respect to the principal rolling directions. The Ti-6Al-4V-0.1B alloy shows an improved rolling response compared with the alloy Ti-6Al-4V because of smaller alpha lamellae size, coherency of alpha/beta interfaces, and multiple slip due to orientation factors. Accelerated dynamic globularization for this alloy is similarly caused by the intralamellar transverse boundary formation via multiple slip and strain accumulation at TiB particles. The (0002)(alpha) pole figures of rolled Ti-6Al-4V alloy shows ``TD splitting'' at lower rolling temperatures because of strong initial texture. Substantial beta phase mitigates the effect of starting texture at higher temperature so that ``RD splitting'' characterizes the basal pole figure. Weak starting texture and easy slip transfer for Ti-6Al-4V-0.1B alloy produce simultaneous TD and RD splittings in basal pole figures at all rolling temperatures.

Microstructure and texture evolution during sub-transus thermo-mechanical processing of Ti-6Al-4V-0.1B alloy: part II. static annealing in (alpha plus beta) regime

The first part of this study describes the evolution of microstructure and texture in Ti-6Al-4V-0.1B alloy during sub-transus rolling vis-A -vis the control alloy Ti-6Al-4V. In the second part, the static annealing response of the two alloys at self-same conditions is compared and the principal micromechanisms are analyzed. Faster globularization kinetics has been observed in the Ti-6Al-4V-0.1B alloy for equivalent annealing conditions. This is primarily attributed to the alpha colonies, which leads to easy boundary splitting via multiple slip activation in this alloy. The other mechanisms facilitating lamellar to equiaxed morphological transformations, e.g., termination migration and cylinderization, also start early in the boron-modified alloy due to small alpha colony size, small aspect ratio of the alpha lamellae, and the presence of TiB particles in the microstructure. Both the alloys exhibit weakening of basal fiber (ND||aOE (c) 0001 >) and strengthening of prism fiber (RD||aOE (c) aOE(a)) upon annealing. A close proximity between the orientations of fully globularized primary alpha and secondary alpha phases during alpha -> beta -> alpha transformation has accounted for such a texture modification.

Effect of temperature and strain rate on the deformation behavior and microstructure of a homogenized AZ31 magnesium alloy

The plastic deformation behavior and dynamic recrystallization (DRX) in homogenized AZ31 Mg alloy was investigated in uniaxial compression in the temperature range between 150 and 400 degrees C with strain rates ranging from 10(-3) to 10(2) s(-1). Twinning was found to contribute significantly during the early stages of deformation. The onset of twinning was examined in detail by recourse to the examination of the appearance of first local maxima before peak strain in the stress-strain responses and the second derivative of stress with strain. High strain hardening rate was observed immediately after the onset of twinning and was found to increase with the Zener-Hollomon parameter. DRX was observed at temperatures above 250 degrees C whereas deformation at lower temperatures (< 250 degrees C) leads to extensive twinning at all the strain rates. At intermediate temperatures of 250-300 degrees C, plastic strains tend to localize near grain/twin boundaries, confining DRX only to these regions. Increase in the temperature promotes non-basal slip, which, in turn, leads to uniform deformation; DRX too becomes uniform. Deformation behavior in three different regimes of temperature is discussed. The dependence of critical stress for the onset of DRX and peak flow stress on temperature and strain rate are also described. (C) 2013 Elsevier B.V. All rights reserved.

Dynamics of the North American continent

The forces that cause deformation of western North America have been debated for decades. Recent studies, primarily based on analysis of crustal stresses in the western United States, have suggested that the deformation of the region is mainly controlled by gravitational potential energy (GPE) variations and boundary loads, with basal tractions due to mantle flow playing a relatively minor role. We address these issues by modelling the deviatoric stress field over western North America from a 3-D finite element mantle circulation model with lateral viscosity variations. Our approach takes into account the contribution from both topography and shallow lithosphere structure (GPE) as well as that from deeper mantle flow in one single model, as opposed to separate lithosphere and circulation models, as has been done so far. In addition to predicting the deviatoric stresses we also jointly fit the constraints of geoid, dynamic topography and plate motion both globally and over North America, in order to ensure that the forces that arise in our models are dynamically consistent. We examine the sensitivity of the dynamic models to different lateral viscosity variations. We find that circulation models that include upper mantle slabs yield a better fit to observed plate velocities. Our results indicate that a model of GPE variations coupled with mantle convection gives the best fit to the observational constraints. We argue that although GPE variations control a large part of the deformation of the western United States, deeper mantle tractions also play a significant role. The average deviatoric stress magnitudes in the western United States range 30-40 MPa. The cratonic region exhibits higher coupling to mantle flow than the rest of the continent. We find that a relatively strong San Andreas fault gives a better fit to the observational constraints, especially that of plate velocity in western North America.

Numerical simulations of CO2 migration during charnockite genesis

Charnockite is considered to be generated either through the dehydration of granitic magma by CO2 purging or by solid-state dehydration through CO2 metasomatism during granulite facies metamorphism. To understand the extent of dehydration, CO2 migration is quantitatively modeled in silicate melt and metasomatic fluid as a function of temperature, H2O wt%, pressure, basal CO2 flux and dynamic viscosity. Numerical simulations show that CO2 advection through porous and permeable high-grade metamorphic rocks can generate dehydrated patches close to the CO2 flow path, as illustrated by the occurrences of ``incipient charnockites.'' CO2 reaction-front velocity constrained by field observations is 0.69 km/m.y., a reasonable value, which matches well with other studies. On the other hand, temperature, rate of cooling, and basal CO2 flux are the critical parameters affecting CO2 diffusion through a silicate melt. CO2 diffusion through silicate melt can only occur at temperature greater than 840 degrees C and during slow cooling (<= 3.7 x 10(-5) degrees C/yr), features that are typical of magma emplacement in the lower crust. Stalling of CO2 fluxing at similar to 840 degrees C explains why some deep-level plutons contain both hydrous and anhydrous (charnockitic) mineral assemblages. CO2 diffusion through silicate melt is virtually insensitive to pressure. Addition of CO2 basal flux facilitates episodic dehydrated melt migration by generating fracture pathways.

Contrasting fire regimes in a seasonally dry tropical forest and a savanna ecosystem in the western Ghats, India

Tropical dry forests and savannas constitute more than half of all tropical forests and grasslands, but little is known about forest fire regimes within these two extensive types of ecosystems. Forest fire regimes in a predominantly dry forest in India, the Nilgiri landscape, and a predominantly savanna ecosystem in the Sathyamangalam landscape, were examined. Remote sensing data were applied to delineate burned areas, determine fire size characteristics, and to estimate fire-rotation intervals. Belt transects (0.5 ha) were used to estimate forest structure, diversity, and fuel loads. Mean area burned, mean number of fires, and mean fire size per year were substantially higher in the Nilgiri landscape compared to the Sathyamangalam landscape. Mean fire-rotational interval was 7.1 yr in the Nilgiri landscape and 44.1 yr in the Sathyamangalam landscape. Tree (>= 10 cm diameter at breast height) species diversity, tree density, and basal area were significantly higher in the Nilgiri landscape compared to the Sathyamangalam landscape. Total fuel loads were significantly higher in tropical dry and moist deciduous forests in the Nilgiri landscape, but total fuel loads were higher in the tropical dry thorn forests of the Sathyamangalam landscape. Thus, the two landscapes revealed contrasting fire regimes and forest characteristics, with more and four-fold larger fires in the Nilgiri landscape. The dry forests and savannas could be maintained by a combination of factors, such as fire, grazing pressures, and herbivore populations. Understanding the factors maintaining these two ecosystems will be critical for their conservation.

Effect of Texture and Grain Size on Bio-Corrosion Response of Ultrafine-Grained Titanium

The bio-corrosion response of ultrafine-grained commercially pure titanium processed by different routes of equal-channel angular pressing has been studied in simulated body fluid. The results indicate that the samples processed through route B-c that involved rotation of the workpiece by 90 deg in the same sense between each pass exhibited higher corrosion resistance compared to the ones processed by other routes of equal-channel angular pressing, as well as the coarse-grained sample. For a similar grain size, the higher corrosion resistance of the samples exhibiting off-basal texture compared to shear texture indicates the major role of texture in corrosion behavior. It is postulated that an optimum combination of microstructure and crystallographic texture can lead to high strength and excellent corrosion resistance.

Long-Term Environmental Correlates of Invasion by Lantana camara (Verbenaceae) in a Seasonally Dry Tropical Forest

Invasive species, local plant communities and invaded ecosystems change over space and time. Quantifying this change may lead to a better understanding of the ecology and the effective management of invasive species. We used data on density of the highly invasive shrub Lantana camara (lantana) for the period 1990-2008 from a 50 ha permanent plot in a seasonally dry tropical forest of Mudumalai in southern India. We used a cumulative link mixed-effects regression approach to model the transition of lantana from one qualitative density state to another as a function of biotic factors such as indicators of competition from local species (lantana itself, perennial grasses, invasive Chromolaena odorata, the native shrub Helicteres isora and basal area of native trees) and abiotic factors such as fire frequency, inter-annual variability of rainfall and relative soil moisture. The density of lantana increased substantially during the study period. Lantana density was negatively associated with the density of H. isora, positively associated with basal area of native trees, but not affected by the presence of grasses or other invasive species. In the absence of fire, lantana density increased with increasing rainfall. When fires occurred, transitions to higher densities occurred at low rainfall values. In drier regions, lantana changed from low to high density as rainfall increased while in wetter regions of the plot, lantana persisted in the dense category irrespective of rainfall. Lantana seems to effectively utilize resources distributed in space and time to its advantage, thus outcompeting local species and maintaining a population that is not yet self-limiting. High-risk areas and years could potentially be identified based on inferences from this study for facilitating management of lantana in tropical dry forests.

Experimental study of fracture behavior of magnesium single crystals

In this work, the fracture behavior of magnesium single crystals is studied by conducting experiments with notched three point bend specimens of three crystallographic orientations. In the first and second orientations, the c-axis is along the normal to the flat surface of the notch, while in the third it is aligned with the notch front. For all the orientations, in situ electron back scattered diffraction observations made around the notch root show profuse tensile twinning of {10 (1) over bar2} type. Further, in the first two orientations basal and prismatic slip traces are identified from optical metallography. The width of the most prominent twin saturates at around 120-150 mu m, while twins continue to nucleate farther away to accommodate plastic deformation. In all the orientations, crack initiation occurs before the attainment of peak load and the crack grows stably along twin-matrix interface before deflecting at twin-twin intersections. Results show that profuse tensile twinning is an important energy dissipating mechanism that enhances the fracture toughness. (C) 2013 Elsevier B.V. All rights reserved.