Design and development of a self-supported polymer electrolyte fuel cell system with anodic dead-end operation

Design and operational details for a self-supported polymer electrolyte fuel cell (PEFC) system with anodic dead-end fuel supply and internally humidified cathodic oxidant flow are described. During the PEFC operation, nitrogen and water back diffuse across the Nafion membrane from the cathode to the anode and accumulate in the anode flow channels affecting stack performance. The accumulated inert species are flushed from the stack by purging the fuel cell stack with a timer-activated purge valve to address the aforesaid problem. To minimize the system complexity, stack is designed in such a way that all the inert species accumulate in only one cell called the purge cell. A pulsed purge sequence comprises opening the valve for purge duration followed by purge-valve closing for the hold period and repeating the sequence in cycles. Since self-humidification is inadequate to keep the membrane wet, the anodic dead-end-operated PEFC stack with composite membrane comprising perflourosulphonic acid (Nafion) and silica is employed for keeping the membrane humidified even while operating the stack with dry hydrogen and internally humidified air.

Structural proteins of mycobacteriophage I3: cloning, expression and sequence analysis of a gene encoding a 70-kDa structural protein

The structural proteins of mycobacteriophage I3 have been analysed by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis (SDS-PAGE), radioiodination and immunoblotting. Based on their abundance the 34- and 70-kDa bands appeared to represent the major structural proteins. Successful cloning and expression of the 70-kDa protein-encoding gene of phage I3 in Escherichia coli and its complete nucleotide sequence determination have been accomplished, A second (partial) open reading frame following the stop codon for the 70-kDa protein was also identified within the cloned fragment. The deduced amino-acid sequence of the 70-kDa protein and the codon usage patterns indicated the preponderance of codons, as predicted from the high G+C content of the genomic DNA of phage I3.

Design and structural analysis of hairpin-TFO for transcriptional activation of genes in S-cerevisiae

Triplex forming oligonucleotides (TFOs) have the potential to modulate gene expression. While most of the experiments are directed towards triplex mediated inhibition of gene expression the strategy potentially could be used for gene specific activation. In an attempt to design a strategy for gene specific activation in vivo applicable to a large number of genes we have designed a TFO based activator-target system which may be utilized in Saccharomyces cerevisiae or any other system where Gal4 protein is ectopically expressed. The total genome sequence of Saccharomyces cerevisiae and expression profiles were used to select the target genes with upstream poly (pu/py) sequences. We have utilized the paradigm of Gal4 protein and its binding site. We describe here the selection of target genes and design of hairpin-TFO including the targeting sequences containing polypurine stretch found in the upstream promoter regions of weakly expressed genes. We demonstrate, the formation of hairpin-TFO, its binding to Gal4 protein, its ability to form triplex with the target duplex in vitro, the effect of polyethylenimine on complex formation and discuss the implication on in vivo transcription activation.

Densification and microstructure development in spark plasma sintered WC–6 wt% ZrO2 nanocomposites

In this paper, we report the results of a transmission electron microscopy investigation on WC–6 wt% ZrO2nanocomposite, spark plasma sintered at 1300 °C, for varying times of up to 20 min. The primary aim of this work was to understand the evolution of microstructure during such a sintering process. The investigation revealed the presence of nanocrystalline ZrO2particles (30–50 nm) entrapped within submicron WC grains. In addition, relatively coarser ZrO2(60–100 nm) particles were observed to be either attached to WC grain boundaries or located at WC triple grain junctions. The evidence of the presence of a small amount of W2C, supposed to have been formed due to sintering reaction between WC and ZrO2, is presented here. Detailed structural investigation indicated that ZrO2in the spark plasma sintered nanocomposite adopted an orthorhombic crystal structure, and the possible reasons for o-ZrO2formation are explained. The increase in kinetics of densification due to the addition of ZrO2is believed to be caused by the enhanced diffusion kinetics in the presence of nonstoichiometric nanocrystalline ZrO2.

Identification of a core promoter and a novel isoform of the human TSCI gene transcript and structural comparison with mouse homolog

Tuberous sclerosis complex (TSC) is an autosomal dominant disorder with loci on chromosome 9q34.12 (TSC1) and chromosome 16p13.3 (TSC2). Genes for both loci have been isolated and characterized. The promoters of both genes have not been characterized so far and little is known about the regulation of these genes. This study reports the characterization of the human TSC1 promoter region for the first time. We have identified a novel alternative isoform in the 5' untranslated region (UTR) of the TSC1 gene transcript involving exon 1. Alternative isoforms in the 5' UTR of the mouse Tsc1 gene transcript involving exon I and exon 2 have also been identified. We have identified three upstream open reading frames (uORFs) in the 5' UTR of the TSC1/Tsc1 gene. A comparative study of the 5' UTR of TSC1/Tsc1 gene has revealed that there is a high degree of similarity not only in the sequence but also in the splicing pattern of both human and mouse TSC1 genes. We have used PCR methodology to isolate approximately 1.6 kb genomic DNA 5' to the TSC1 cDNA. This sequence has directed a high level of expression of luciferase activity in both HeLa and HepG2 cells. Successive 5' and 3' deletion analysis has suggested that a -587 bp region, from position +77 to -510 from the transcription start site (TSS), contains the promoter activity. Interestingly, this region contains no consensus TATA box or CAAT box. However, a 521-bp fragment surrounding the TSS exhibits the characteristics of a CpG island which overlaps with the promoter region. The identification of the TSC1 promoter region will help in designing a suitable strategy to identify mutations in this region in patients who do not show any mutations in the coding regions. It will also help to study the regulation of the TSC1 gene and its role in tumorigenesis. (C) 2003 Elsevier B.V. All rights reserved.

The biological and structural characterization of Mycobacterium tuberculosis UvrA provides novel insights into its mechanism of action

Mycobacterium tuberculosis is an extremely well adapted intracellular human pathogen that is exposed to multiple DNA damaging chemical assaults originating from the host defence mechanisms. As a consequence, this bacterium is thought to possess highly efficient DNA repair machineries, the nucleotide excision repair (NER) system amongst these. Although NER is of central importance to DNA repair in M. tuberculosis, our understanding of the processes in this species is limited. The conserved UvrABC endonuclease represents the multi-enzymatic core in bacterial NER, where the UvrA ATPase provides the DNA lesion-sensing function. The herein reported genetic analysis demonstrates that M. tuberculosis UvrA is important for the repair of nitrosative and oxidative DNA damage. Moreover, our biochemical and structural characterization of recombinant M. tuberculosis UvrA contributes new insights into its mechanism of action. In particular, the structural investigation reveals an unprecedented conformation of the UvrB-binding domain that we propose to be of functional relevance. Taken together, our data suggest UvrA as a potential target for the development of novel anti-tubercular agents and provide a biochemical framework for the identification of small-molecule inhibitors interfering with the NER activity in M. tuberculosis.

Recent Research in Structural Health Monitoring

Roctest Group believes in the importance of maintaining a close contact with the scientific community active in fields close to our activities domains, in particular smart structures, structural engineering, sensing and fiber optic sensors. These contacts allow Roctest SMARTEC Telemac to remain at the front of scientific progress and to contribute to the diffusion of the monitoring culture worldwide. Our research and development team actively contributes in the research community, attending conferences and regularly publishing in the scientific literature. we support academic research by participating in joint research projects and by regularly welcoming graduate and undergraduate students for stages and exchange programs.

Structural, electrical, and optical properties of as-grown and heat treated ultra-thin SnS films

The composition, structural, electrical, and optical properties of as-grown and heat treated tin-mono-sulfide (SnS) ultra-thin films have been studied. The ultra-thin SnS films were prepared on glass substrates by thermal resistive evaporation technique. All the SnS films contained nanocrystallites and exhibited p-type conductivity with a low Hall-mobility, <50 cm(2)/Vs. All these films are highly tin rich in nature and exhibited orthorhombic crystal structure. As compared to other films, the SnS films annealed at 300 degrees C showed a low electrical resistivity of similar to 36 Omega cm with an optical band gap of similar to 1.98 eV. The observed electrical and optical properties of all the films are discussed based on their composition and structural parameters. These nanocrystalline ultra-thin SnS films could be expected as a buffer layer for the development of tandem solar cell devices due to their low-resistivity and high absorbability with an optimum band gap. (C) 2011 Elsevier B.V. All rights reserved.

Transcription factor erect wing (EWG) is involved in indirect flight muscle patterning, development and maintenance in Drosophila

Muscle development is a multistep process which includes myoblast diversification, proliferation, migration, fusion, differentiation and growth. A hierarchical exhibition of myogenic factors is important for dexterous execution of progressive events in muscle formation. EWG (erect wing) is a transcription factor known to have a role in indirect flight muscle development (IFM) in Drosophila. We marked out the precise spatio-temporal expression profile of EWG in the myoblasts, and in the developing muscles. Mutant adult flies null for EWG in myoblasts show variable number of IFM, suggesting that EWG is required for patterning of the IFM. The remnant muscle found in the EWG null flies show proper assembly of the structural proteins, which implies that some myoblasts manage to fuse, develop and differentiate normally indicating that EWG is not required for differentiation program per se. However, when EWG expression is extended beyond its expression window in a wild type background, muscle thinning is observed implying EWG function in protein synthesis inhibition. Mis-expression studies in wing disc myoblasts hinted at its role in myoblast proliferation. We thus conclude that EWG is important for regulating fusion events which in turn decides the IFM pattern. Also IFM in EWG null mutants show clumps containing broken fibres and an altered mitochondrial morphology. The vertebrate homolog of EWG is nuclear respiratory factor1 (NRF1) which is known to have a function in mitochondrial biogenesis and protection against oxidative stress. Gene expression for inner mitochondrial membrane protein, Opa1-like was found to be absent in these mutants. Also, these flies were more sensitive to oxidative stress, indicating a compromised mitochondrial functioning. Our results therefore demonstrate that EWG functions in maintaining muscles’ structural integrity by ensuing proper mitochondrial activity.

Development of uniformly grown ZnO NPs films using single precursor solution by pulsed spray pyrolysis technique

In this article, we have reported the controlled synthesis of uniformly grown zinc oxide nanoparticles (ZnO NPs) films by a simple, low-cost, and scalable pulsed spray pyrolysis technique. From the surface analysis it is noticed that the as-deposited films have uniformly dispersed NPs-like morphology. The structural studies reveal that these NPs films have highly crystalline hexagonal crystal structure, which are preferentially orientated along the (001) planes. The size of the NPs varied between 5 and 100 nm, and exhibited good stoichiometric chemical composition. Raman spectroscopic analysis reveals that these ZnO NPs films have pure single phase and hexagonal crystal structure. These unique nanostructured films exhibited a low electrical resistivity (5 Omega cm) and high light transmittance (90 %) in visible region.

Structural Integrity of the Greek Key Motif in beta gamma-Crystallins Is Vital for Central Eye Lens Transparency

Background: We highlight an unrecognized physiological role for the Greek key motif, an evolutionarily conserved super-secondary structural topology of the beta gamma-crystallins. These proteins constitute the bulk of the human eye lens, packed at very high concentrations in a compact, globular, short-range order, generating transparency. Congenital cataract (affecting 400,000 newborns yearly worldwide), associated with 54 mutations in beta gamma-crystallins, occurs in two major phenotypes nuclear cataract, which blocks the central visual axis, hampering the development of the growing eye and demanding earliest intervention, and the milder peripheral progressive cataract where surgery can wait. In order to understand this phenotypic dichotomy at the molecular level, we have studied the structural and aggregation features of representative mutations. Methods: Wild type and several representative mutant proteins were cloned, expressed and purified and their secondary and tertiary structural details, as well as structural stability, were compared in solution, using spectroscopy. Their tendencies to aggregate in vitro and in cellulo were also compared. In addition, we analyzed their structural differences by molecular modeling in silico. Results: Based on their properties, mutants are seen to fall into two classes. Mutants A36P, L45PL54P, R140X, and G165fs display lowered solubility and structural stability, expose several buried residues to the surface, aggregate in vitro and in cellulo, and disturb/distort the Greek key motif. And they are associated with nuclear cataract. In contrast, mutants P24T and R77S, associated with peripheral cataract, behave quite similar to the wild type molecule, and do not affect the Greek key topology. Conclusion: When a mutation distorts even one of the four Greek key motifs, the protein readily self-aggregates and precipitates, consistent with the phenotype of nuclear cataract, while mutations not affecting the motif display `native state aggregation', leading to peripheral cataract, thus offering a protein structural rationale for the cataract phenotypic dichotomy ``distort motif, lose central vision''.

Eu3+-activated SrMoO4 phosphors for white LEDs applications: Synthesis and structural characterization

We report the synthesis of Eu3+-activated SrMoO4 phosphors by the facile nitrate-citrate gel combustion method. Powder XRD and Rietveld refinement data confirmed that these phosphors have a monophasic scheelite-type tetragonal structure with space group I4(1)/a (No. 88). FESEM micrographs indicate the agglomerated spherical particles. FTIR spectra showed four stretching and bending vibrational modes (2A(u) and 2E(u)). UV-Visible absorption spectroscopy illustrated that the optical band gap energy (E-g) values increase with increase in Eu3+ concentration. The host SrMoO4 phosphor exhibited an intense blue emission under UV excitation (368 nm). The Eu3+-activated SrMoO4 phosphors revealed characteristic luminescence due to Eu3+ ion corresponding to D-5(1) -> F-7(J) (J = 1,2) and D-5(0) -> F-7(J) (J = 1,2,3,4) transitions upon 465 nm excitation. The electric dipole transition located at 615 nm (D-5(0) -> F-7(2)) was stronger than the magnetic dipole transition located at 592 nm (D-5(0) -> F-7(1)). Intensity parameters (Omega(2), Omega(4)) and radiative properties such as transition probabilities (A(T)), radiative lifetime (tau(rad)) and branching ratio (beta) of Eu3+-activated SrMoO4 phosphors were calculated using the Judd-Ofelt theory. Based on the CIE chromaticity diagram, these phosphors can be promising materials for the development of blue and orange-red component in white LEDs. (C) 2015 Elsevier B.V. All rights reserved.

Metallophosphoesterases: structural fidelity with functional promiscuity

Calcineurin-like metallophosphoesterases (MPEs) form a large superfamily of binuclear metal-ion-centre-containing enzymes that hydrolyse phosphomono-, phosphodi-or phosphotri-esters in a metal-dependent manner. The MPE domain is found in Mre11/SbcD DNA-repair enzymes, mammalian phosphoprotein phosphatases, acid sphingomyelinases, purple acid phosphatases, nucleotidases and bacterial cyclic nucleotide phosphodiesterases. Despite this functional diversity, MPEs show a remarkably similar structural fold and active-site architecture. In the present review, we summarize the available structural, biochemical and functional information on these proteins. We also describe how diversification and specialization of the core MPE fold in various MPEs is achieved by amino acid substitution in their active sites, metal ions and regulatory effects of accessory domains. Finally, we discuss emerging roles of these proteins as non-catalytic protein-interaction scaffolds. Thus we view the MPE superfamily as a set of proteins with a highly conserved structural core that allows embellishment to result in dramatic and niche-specific diversification of function.

Cyclic nucleotide binding and structural changes in the isolated GAF domain of Anabaena adenylyl cyclase, CyaB2

GAF domains are a large family of regulatory domains, and a subset are found associated with enzymes involved in cyclic nucleotide (cNMP) metabolism such as adenylyl cyclases and phosphodiesterases. CyaB2, an adenylyl cyclase from Anabaena, contains two GAF domains in tandem at the N-terminus and an adenylyl cyclase domain at the C-terminus. Cyclic AMP, but not cGMP, binding to the GAF domains of CyaB2 increases the activity of the cyclase domain leading to enhanced synthesis of cAMP. Here we show that the isolated GAFb domain of CyaB2 can bind both cAMP and cGMP, and enhanced specificity for cAMP is observed only when both the GAFa and the GAFb domains are present in tandem(GAFab domain). In silico docking and mutational analysis identified distinct residues important for interaction with either cAMP or cGMP in the GAFb domain. Structural changes associated with ligand binding to the GAF domains could not be detected by bioluminescence resonance energy transfer (BRET) experiments. However, amide hydrogen-deuterium exchange mass spectrometry (HDXMS) experiments provided insights into the structural basis for cAMP-induced allosteric regulation of the GAF domains, and differences in the changes induced by cAMP and cGMP binding to the GAF domain. Thus, our findings could allow the development of molecules that modulate the allosteric regulation by GAF domains present in pharmacologically relevant proteins.