Src Homology 3-interacting Domain of Rv1917c of Mycobacterium tuberculosis Induces Selective Maturation of Human Dendritic Cells by Regulating PI3K-MAPK-NF-kappa B Signaling and Drives Th2 Immune Responses

Mycobacterium tuberculosis, an etiological agent of pulmonary tuberculosis, causes significant morbidity and mortality worldwide. Pathogenic mycobacteria survive in the host by subverting host innate immunity. Dendritic cells (DCs) are professional antigen-presenting cells that are vital for eliciting immune responses to infectious agents, including pathogenic mycobacteria. DCs orchestrate distinct Th responses based on the signals they receive. In this perspective, deciphering the interactions of the proline-glutamic acid/proline-proline-glutamic acid (PE/PPE) family of proteins of M. tuberculosis with DCs assumes significant pathophysiological attributes. In this study, we demonstrate that Rv1917c (PPE34), a representative member of the proline-proline-glutamic-major polymorphic tandem repeat family, interacts with TLR2 and triggers functional maturation of human DCs. Signaling perturbations implicated a critical role for integrated cross-talk among PI3K-MAPK and NF-kappa B signaling cascades in Rv1917c-induced maturation of DCs. However, this maturation of DCs was associated with a secretion of high amounts of anti-inflammatory cytokine IL-10, whereas Th1-polarizing cytokine IL-12 was not induced. Consistent with these results, Rv1917c-matured DCs favored secretion of IL-4, IL-5, and IL-10 from CD4(+) T cells and contributed to Th2-skewed cytokine balance ex vivo in healthy individuals and in patients with pulmonary tuberculosis. Interestingly, the Rv1917c-skewed Th2 immune response involved induced expression of cyclooxygenase-2 (COX-2) in DCs. Taken together, these results indicate that Rv1917c facilitates a shift in the ensuing immunity toward the Th2 phenotype and could aid in immune evasion by mycobacteria.

Influence of Osmotic Suction on the Soil-Water Characteristic Curves of Compacted Expansive Clay

Unsaturated clays are subject to osmotic suction gradients in geoenvironmental engineering applications and it therefore becomes important to understand the effect of these chemical concentration gradients on soil-water characteristic curves (SWCCs). This paper brings out the influence of induced osmotic suction gradient on the wetting SWCCs of compacted clay specimens inundated with sodium chloride solutions/distilled water at vertical stress of 6.25 kPa in oedometer cells. The experimental results illustrate that variations in initial osmotic suction difference induce different magnitudes of osmotic induced consolidation and osmotic consolidation strains thereby impacting the wetting SWCCs and equilibrium water contents of identically compacted clay specimens. Osmotic suction induced by chemical concentration gradients between reservoir salt solution and soil-water can be treated as an equivalent net stress component, (p(pi)) that decreases the swelling strains of unsaturated specimens from reduction in microstructural and macrostructural swelling components. The direction of osmotic flow affects the matric SWCCs. Unsaturated specimens experiencing osmotic induced consolidation and osmotic consolidation develop lower equilibrium water content than specimens experiencing osmotic swelling during the wetting path. The findings of the study illustrate the need to incorporate the influence of osmotic suction in determination of the matric SWCCs.

Heat Shock Protein 90 as a Drug Target against Protozoan Infections Biochemical characterization of HSP90 From plasmodium falciparum and trypanosoma evansi and evaluation of its inhibitor as a candidate drug

Using a pharmacological inhibitor of Hsp90 in cultured malarial parasite, we have previously implicated Plasmodium falciparum Hsp90 (PfHsp90) as a drug target against malaria. In this study, we have biochemically characterized PfHsp90 in terms of its ATPase activity and interaction with its inhibitor geldanamycin (GA) and evaluated its potential as a drug target in a preclinical mouse model of malaria. In addition, we have explored the potential of Hsp90 inhibitors as drugs for the treatment of Trypanosoma infection in animals. Our studies with full-length PfHsp90 showed it to have the highest ATPase activity of all known Hsp90s; its ATPase activity was 6 times higher than that of human Hsp90. Also, GA brought about more robust inhibition of PfHsp90 ATPase activity as compared with human Hsp90. Mass spectrometric analysis of PfHsp90 expressed in P. falciparum identified a site of acetylation that overlapped with Aha1 and p23 binding domain, suggesting its role in modulating Hsp90 multichaperone complex assembly. Indeed, treatment of P. falciparum cultures with a histone deacetylase inhibitor resulted in a partial dissociation of PfHsp90 complex. Furthermore, we found a well known, semisynthetic Hsp90 inhibitor, namely 17-(allylamino)-17-demethoxygeldanamycin, to be effective in attenuating parasite growth and prolonging survival in a mouse model of malaria. We also characterized GA binding to Hsp90 from another protozoan parasite, namely Trypanosoma evansi. We found 17-(allylamino)-17-demethoxygeldanamycin to potently inhibit T. evansi growth in a mouse model of trypanosomiasis. In all, our biochemical characterization, drug interaction, and animal studies supported Hsp90 as a drug target and its inhibitor as a potential drug against protozoan diseases.

Defective in Cuticular Ridges (DCR) of Arabidopsis thaliana, a Gene Associated with Surface Cutin Formation, Encodes a Soluble Diacylglycerol Acyltransferase

A key step in the triacylglycerol (TAG) biosynthetic pathway is the final acylation of diacylglycerol (DAG) by DAG acyltransferase. In silico analysis has revealed that the DCR (defective in cuticular ridges) (At5g23940) gene has a typical HX4D acyltransferase motif at the N-terminal end and a lipid binding motif VX(2)GF at the middle of the sequence. To understand the biochemical function, the gene was overexpressed in Escherichia coli, and the purified recombinant protein was found to acylate DAG specifically in an acyl-CoA-dependent manner. Overexpression of At5g23940 in a Saccharomyces cerevisiae quadruple mutant deficient in DAG acyltransferases resulted in TAG accumulation. At5g23940 rescued the growth of this quadruple mutant in the oleate-containing medium, whereas empty vector control did not. Lipid particles were localized in the cytosol of At5g23940-transformed quadruple mutant cells, as observed by oil red O staining. There was an incorporation of 16-hydroxyhexadecanoic acid into TAG in At5g23940-transformed cells of quadruple mutant. Here we report a soluble acyl-CoA-dependent DAG acyltransferase from Arabidopsis thaliana. Taken together, these data suggest that a broad specific DAG acyltransferase may be involved in the cutin as well as in the TAG biosynthesis by supplying hydroxy fatty acid.

Detrimental Effects of Hypoxia-Specific Expression of Uracil DNA Glycosylase (Ung) in Mycobacterium smegmatis

Mycobacterium tuberculosis is known to reside latently in a significant fraction of the human population. Although the bacterium possesses an aerobic mode of metabolism, it adapts to persistence under hypoxic conditions such as those encountered in granulomas. While in mammalian systems hypoxia is a recognized DNA-damaging stress, aspects of DNA repair in mycobacteria under such conditions have not been studied. We subjected Mycobacterium smegmatis, a model organism, to the Wayne's protocol of hypoxia. Analysis of the mRNA of a key DNA repair enzyme, uracil DNA glycosylase (Ung), by real-time reverse transcriptase PCR (RT-PCR) revealed its downregulation during hypoxia. However, within an hour of recovery of the culture under normal oxygen levels, the Ung mRNA was restored. Analysis of Ung by immunoblotting and enzyme assays supported the RNA analysis results. To understand its physiological significance, we misexpressed Ung in M. smegmatis by using a hypoxia-responsive promoter of narK2 from M. tuberculosis. Although the misexpression of Ung during hypoxia decreased C-to-T mutations, it compromised bacterial survival upon recovery at normal oxygen levels. RT-PCR analysis of other base excision repair gene transcripts (UdgB and Fpg) suggested that these DNA repair functions also share with Ung the phenomenon of downregulation during hypoxia and recovery with return to normal oxygen conditions. We discuss the potential utility of this phenomenon in developing attenuated strains of mycobacteria.

A PEFC With Pt-TiO2/C as Oxygen-Reduction Catalyst

Carbon-supported Pt-TiO2 (Pt-TiO2/C) catalyst with varying atomic ratio of Pt to Ti, namely, 1: 1, 2: 1, and 3: 1, is prepared by sol-gel method and its electrocatalytic activity toward oxygen-reduction reaction (ORR) is evaluated for the application in polymer electrolyte fuel cells (PEFCs). The optimum atomic ratio of Pt to Ti in Pt-TiO2/C and annealing temperature are established by cyclic voltammetry and fuel-cell-polarization studies. Pt-TiO2/C annealed at 750 degrees C with Pt and Ti in atomic ratio of 2: 1, namely, 750 Pt-TiO2/C (2: 1), shows enhanced electrocatalytic activity toward ORR. It is found that the incorporation of TiO2 with Pt ameliorates both electrocatalytic activity and stability of cathode in relation to pristine Pt cathode, currently being used in PEFCs. A power density of 0.75 W/cm(2) is achieved at 0.6 V for the PEFC with 750 Pt-TiO2/C (2: 1) as compared with 0.62 W/cm(2) at 0.6 V achieved with the PEFC comprising Pt/C as cathode catalyst while operating under identical conditions. Interestingly, carbon-supported Pt-TiO2 cathode exhibits only 6% loss in electrochemical surface area after 5000 potential cycles while it is as high as 25% for Pt/C. DOI: 10.1115/1.4002466]

Unified Galerkin- and DAE-Based Approximation of Fractional Order Systems

We consider numerical solutions of nonlinear multiterm fractional integrodifferential equations, where the order of the highest derivative is fractional and positive but is otherwise arbitrary. Here, we extend and unify our previous work, where a Galerkin method was developed for efficiently approximating fractional order operators and where elements of the present differential algebraic equation (DAE) formulation were introduced. The DAE system developed here for arbitrary orders of the fractional derivative includes an added block of equations for each fractional order operator, as well as forcing terms arising from nonzero initial conditions. We motivate and explain the structure of the DAE in detail. We explain how nonzero initial conditions should be incorporated within the approximation. We point out that our approach approximates the system and not a specific solution. Consequently, some questions not easily accessible to solvers of initial value problems, such as stability analyses, can be tackled using our approach. Numerical examples show excellent accuracy. DOI: 10.1115/1.4002516]

Mechanical Properties of Al-Mica Particulate Composite Material

Cast aluminium alloy mica particle composites of varying mica content were tested in tension, compression, and impact. With 2.2 percent mica (size range 40µm – 120µm) the tensile and compression strengths of aluminium alloy decreased by 56 and 22 percent, respectively. The corresponding decreases in percent elongation and percent reduction are 49 and 39 percent. Previous work [2] shows that despite this decrease in strength the composite with 2.5 percent mica and having an UTS of 15 kg/mm2 and compression strength of 28 kg/mm2 performs well as a bearing material under severe running conditions. The differences in strength characteristics of cast aluminium-mica particle composites between tension and compression suggests that, as in cast iron, expansion of voids at the matrix particle interface may be the guiding mechanism of the deformation. SEM studies show that on the tensile fractured specimen surface, there are large voids at the particle matrix interface.

A Bilinear Constitutive Model for Isotropic Bimodulus Materials

Proper formulation of stress-strain relations, particularly in tension-compression situations for isotropic biomodulus materials, is an unresolved problem. Ambartsumyan's model [8] and Jones' weighted compliance matrix model [9] do not satisfy the principle of coordinate invariance. Shapiro's first stress invariant model [10] is too simple a model to describe the behavior of real materials. In fact, Rigbi [13] has raised a question about the compatibility of bimodularity with isotropy in a solid. Medri [2] has opined that linear principal strain-principal stress relations are fictitious, and warned that the bilinear approximation of uniaxial stress-strain behavior leads to ill-working bimodulus material model under combined loading. In the present work, a general bilinear constitutive model has been presented and described in biaxial principal stress plane with zonewise linear principal strain-principal stress relations. Elastic coefficients in the model are characterized based on the signs of (i) principal stresses, (ii) principal strains, and (iii) on the value of strain energy component ratio ER greater than or less than unity. The last criterion is used in tension-compression and compression-tension situations to account for different shear moduli in pure shear stress and pure shear strain states as well as unequal cross compliances.

Analysis of Compressibility of Sensitive Soils

Sensitive soils, in general, are prone to mechanical disturbances while sampling, handling, and testing. This necessitates the prediction of true field behavior. The compressibility response of such soils is typical of having three zones, mechanistically explained as nonparticulate, transitional, and particulate. Such zoning has enabled the development of a simple method to predict the field compressibility response of the sample. The field compression curve with sigmact act as the most probable yield stress is considered to reflect 0% disturbance. By a comparison of experimentally determined sigmac and sigmact, it is possible to estimate the degree of sample disturbance. When the value of sigmac is closer to sigmact, the sampling disturbance approaches zero. As the value of sigmac reduces, the degree of sampling disturbance increases. The possibility of using this degree of sample disturbance from compressibility data to obtain other true properties from laboratory results of the sampled specimens has been examined.

Ultimate Strength of RC Wall Panels with Openings

Test results of 12 reinforced concrete (RC) wall panels with openings are presented. The panels have been subjected to in-plane vertical loads applied at an eccentricity to represent possible accidental eccentricity that occurs in practice due to constructional imperfections. The 12 specimens consist of two identical groups of six panels each. One group of panels is tested in one-way in-plane action (i.e., supported at top and bottom edges against lateral displacement). The second group of panels is tested in two-way in-plane action (i.e., supported on all the four edges against lateral displacement). Openings in the panels represent typical door and window openings. Cracking loads, ultimate loads, crack patterns, and lateral deflections of the panels are studied. Empirical methods have been developed for the prediction of ultimate load. Also, lateral deflections, cracking loads, and ultimate loads of identical loads tested under one-way and two-way action are compared.

Ultimate Strength of R.C. Wall Panels in Two-Way In-Plane Action

Test results of 24 reinforced concrete wall panels in two-way action (i.e., supported on all the four sides) and subjected to in-plane vertical load are presented. The load is applied at an eccentricity to represent possible accidental eccentricity that occurs in practice due to constructional imperfections. Influences of aspect ratio, thinness ratio, slendemess ratio, vertical steel, and horizontal steel on the ultimate load are studied. Two equations are proposed to predict the ultimate load carried by the panels. The first equation is empirical and is arrived at from trial and error fitting with test data. The second equation is semi-empirical and is developed from a modification of the buckling strength of thin rectangular plates. Both the equations are formulated so as to give a safe prediction of a large portion of ultimate strength test results. Also, ultimate load cracking load and lateral deflections of identical panels in two-way action (all four sides supported) and oneway action (top and bottom sides only supported) are compared.