288 resultados para Agrobacterium-mediated transformation
Resumo:
A hitherto unseen rotation of the isopropyl group in the solid state, predicted to be forbidden based on theoretical investigations, is reported. This C-C rotation observed during the temperature dependent single-crystal-to-single-crystal transformation is attributed to the concomitant changes in molecular structure and intermolecular packing.
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Protein lysine acetylation is known to regulate multiple aspects of bacterial metabolism. However, its presence in mycobacterial signal transduction and virulence-associated proteins has not been studied. In this study, analysis of mycobacterial proteins from different cellular fractions indicated dynamic and widespread occurrence of lysine acetylation. Mycobacterium tuberculosis proteins regulating diverse physiological processes were then selected and expressed in the surrogate host Mycobacterium smegmatis. The purified proteins were analyzed for the presence of lysine acetylation, leading to the identification of 24 acetylated proteins. In addition, novel lysine succinylation and propionylation events were found to co-occur with acetylation on several proteins. Protein-tyrosine phosphatase B (PtpB), a secretory phosphatase that regulates phosphorylation of host proteins and plays a critical role in Mycobacterium infection, is modified by acetylation and succinylation at Lys-224. This residue is situated in a lid region that covers the enzyme's active site. Consequently, acetylation and succinylation negatively regulate the activity of PtpB.
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Staphylococcus aureus necrotizing pneumonia is recognized as a toxin-mediated disease, yet the tissue-destructive events remain elusive, partly as a result of lack of mechanistic studies in human lung tissue. In this study, a three-dimensional (3D) tissue model composed of human lung epithelial cells and fibroblasts was used to delineate the role of specific staphylococcal exotoxins in tissue pathology associated with severe pneumonia. To this end, the models were exposed to the mixture of exotoxins produced by S. aureus strains isolated from patients with varying severity of lung infection, namely necrotizing pneumonia or lung empyema, or to purified toxins. The necrotizing pneumonia strains secreted high levels of alpha-toxin and Panton-Valentine leukocidin (PVL), and triggered high cytotoxicity, inflammation, necrosis and loss of E-cadherin from the lung epithelium. In contrast, the lung empyema strain produced moderate levels of PVL, but negligible amounts of alpha-toxin, and triggered limited tissue damage. alpha-toxin had a direct damaging effect on the epithelium, as verified using toxin-deficient mutants and pure alpha-toxin. Moreover, PVL contributed to pathology through the lysis of neutrophils. A combination of alpha-toxin and PVL resulted in the most severe epithelial injury. In addition, toxin-induced release of pro-inflammatory mediators from lung tissue models resulted in enhanced neutrophil migration. Using a collection of 31 strains from patients with staphylococcal pneumonia revealed that strains producing high levels of alpha-toxin and PVL were cytotoxic and associated with fatal outcome. Also, the strains that produced the highest toxin levels induced significantly greater epithelial disruption. Of importance, toxin-mediated lung epithelium destruction could be inhibited by polyspecific intravenous immunoglobulin containing antibodies against alpha-toxin and PVL. This study introduces a novel model system for study of staphylococcal pneumonia in a human setting. The results reveal that the combination and levels of alpha-toxin and PVL correlate with tissue pathology and clinical outcome associated with pneumonia.
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The screen printed electrochemical glucose sensor is developed suitable for revere iontophoresis (RI) application. Glucose oxidase is immobilized on screen printed sensor using crosslinking method. Electrochemical and material characterization studies are conducted on the developed sensor and the obtained results confirm the suitability of the developed sensor for RI application. The developed sensor is validated by conducting clinical investigations on 10 human subjects through RI. A correlation is established between the blood glucose and extracted glucose, and correlation is found to be 0.73. (C) 2015 The Electrochemical Society. All rights reserved.
Resumo:
We present experimental and theoretical results on monolayer colloidal cadmium selenide quantum dot films embedded with tiny gold nanoparticles. By varying the density of the embedded gold nanoparticles, we were able to engineer a plasmon-mediated crossover from emission quenching to enhancement regime at interparticle distances for which only quenching of emission is expected. This crossover and a nonmonotonic variation of photoluminescence intensity and decay rate, in experiments, is explained in terms of a model for plasmon-mediated collective emission of quantum emitters which points to the emergence of a new regime in plasmon-exciton interactions. The presented methodology to achieve enhancement in optical quantum efficiency for optimal doping of gold nanoparticles in such ultrathin high-density quantum dot films can be beneficial for new-generation displays and photodetectors.
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Mechanical behavior of three-dimensional cellular assembly of graphene foam (GF) presented temperature dependent characteristics evaluated at both low temperature and room temperature conditions. Cellular structure of GF comprised of polydimethyl siloxane polymer as a flexible supporting material demonstrated 94% enhancement in the storage modulus as compared to polymer foam alone. Evaluation of frequency dependence revealed an increase in both storage modulus and tan delta with the increase in frequency. Moreover, strain rate independent highly reversible behavior is measured up to several compression cycles at larger strains. It is elucidated that the interaction between graphene and polymer plays a crucial role in thermo-mechanical stability of the cellular structure. (C) 2015 Elsevier Ltd. All rights reserved.
Resumo:
There is great interest in lead-free (Ba0.85Ca0.15)(Ti0.90Zr0.10)O-3 (15/10BCTZ) because of its exceptionally large piezoelectric response Liu and Ren, Phys. Rev. Lett. 103, 257602 (2009)]. In this paper, we have analyzed the nature of: (i) crystallographic phase coexistence at room temperature, (ii) temperature-and field-induced phase transformation to throw light on the atomistic mechanisms associated with the large piezoelectric response of this system. A detailed temperature-dependent dielectric and lattice thermal expansion study proved that the system exhibits a weak dielectric relaxation, characteristic of a relaxor ferroelectric material on the verge of exhibiting a normal ferroelectric-paraelectric transformation. Careful structural analysis revealed that a ferroelectric state at room temperature is composed of three phase coexistences, tetragonal (P4mm)+ orthorhombic (Amm2) + rhombohedral (R3m). We also demonstrate that the giant piezoresponse is associated with a significant fraction of the tetragonal phase transforming to rhombohedral. It is argued that the polar nanoregions associated with relaxor ferroelectricity amplify the piezoresponse by providing an additional degree of intrinsic structural inhomogeneity to the system.
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Mitochondrial DNA (mtDNA) deletions are associated with various mitochondrial disorders. The deletions identified in humans are flanked by short, directly repeated mitochondrial DNA sequences; however, the mechanism of such DNA rearrangements has yet to be elucidated. In contrast to nuclear DNA (nDNA), mtDNA is more exposed to oxidative damage, which may result in double-strand breaks (DSBs). Although DSB repair in nDNA is well studied, repair mechanisms in mitochondria are not characterized. In the present study, we investigate the mechanisms of DSB repair in mitochondria using in vitro and ex vivo assays. Whereas classical NHEJ (C-NHEJ) is undetectable, microhomology-mediated alternative NHEJ efficiently repairs DSBs in mitochondria. Of interest, robust microhomology-mediated end joining (MMEJ) was observed with DNA substrates bearing 5-, 8-, 10-, 13-, 16-, 19-, and 22-nt microhomology. Furthermore, MMEJ efficiency was enhanced with an increase in the length of homology. Western blotting, immunoprecipitation, and protein inhibition assays suggest the involvement of CtIP, FEN1, MRE11, and PARP1 in mitochondrial MMEJ. Knock-down studies, in conjunction with other experiments, demonstrated that DNA ligase III, but not ligase IV or ligase I, is primarily responsible for the final sealing of DSBs during mitochondrial MMEJ. These observations highlight the central role of MMEJ in maintenance of mammalian mitochondrial genome integrity and is likely relevant for deletions observed in many human mitochondrial disorders.
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Gadolinium oxide, cerium oxide, and 10 mol% gadolinia doped ceria ceramic powders have been synthesized using combustion technique. Though the cubic gadolinia phase is stable at room temperature, single phase monoclinic gadolinia was obtained as a result of combustion synthesis using fuel lean and stoichiometric precursor compositions. This powder was subjected to calcination treatment and ceria doping to study the stability of phases and the rate of phase transformation from monoclinic to cubic gadolinia. It was found that monoclinic gadolinia transforms to cubic gadolinia upon calcination at temperatures less than 1200 degrees C. It was also found that rate of phase transformation is more for powder produced using fuel lean compositions; and the rate is enhanced upon ceria doping. (C) 2015 Elsevier Ltd. All rights reserved.
Resumo:
Cross domain and cross-modal matching has many applications in the field of computer vision and pattern recognition. A few examples are heterogeneous face recognition, cross view action recognition, etc. This is a very challenging task since the data in two domains can differ significantly. In this work, we propose a coupled dictionary and transformation learning approach that models the relationship between the data in both domains. The approach learns a pair of transformation matrices that map the data in the two domains in such a manner that they share common sparse representations with respect to their own dictionaries in the transformed space. The dictionaries for the two domains are learnt in a coupled manner with an additional discriminative term to ensure improved recognition performance. The dictionaries and the transformation matrices are jointly updated in an iterative manner. The applicability of the proposed approach is illustrated by evaluating its performance on different challenging tasks: face recognition across pose, illumination and resolution, heterogeneous face recognition and cross view action recognition. Extensive experiments on five datasets namely, CMU-PIE, Multi-PIE, ChokePoint, HFB and IXMAS datasets and comparisons with several state-of-the-art approaches show the effectiveness of the proposed approach. (C) 2015 Elsevier B.V. All rights reserved.
Resumo:
The biodegradation of polycaprolactone (PCL), polylactic acid (PLA), polyglycolide (PGA) and their copolymers, poly (lactide-co-glycolide) and poly (D, L-lactide-co-caprolactone) (PLCL) was investigated. The influence of different solvents on the degradation of these polymers at 37 degrees C in the presence of two different lipases namely Novozym 435 and the free lipase of porcine pancreas was investigated. The rate coefficients for the polymer degradation and enzyme deactivation were determined using continuous distribution kinetics. Among the homopolymers, the degradation of PGA was nearly an order of magnitude lower than that for PCL and PLA. The overall rate coefficients of the copolymers were higher than their respective homopolymers. Thus, PLCL degraded faster than either PCL or PLA. The degradation was highly dependent on the viscosity of the solvent used with the highest degradation observed in acetone. The degradation of the polymers in acetone was nearly twice that observed in dimethyl sulfoxide indicating that the degradation decreases with increase in the solvent viscosity. The degradation of the polymers in water-solvent mixtures indicated an optimal water content of 2.5 wt% of water.
Resumo:
Cross domain and cross-modal matching has many applications in the field of computer vision and pattern recognition. A few examples are heterogeneous face recognition, cross view action recognition, etc. This is a very challenging task since the data in two domains can differ significantly. In this work, we propose a coupled dictionary and transformation learning approach that models the relationship between the data in both domains. The approach learns a pair of transformation matrices that map the data in the two domains in such a manner that they share common sparse representations with respect to their own dictionaries in the transformed space. The dictionaries for the two domains are learnt in a coupled manner with an additional discriminative term to ensure improved recognition performance. The dictionaries and the transformation matrices are jointly updated in an iterative manner. The applicability of the proposed approach is illustrated by evaluating its performance on different challenging tasks: face recognition across pose, illumination and resolution, heterogeneous face recognition and cross view action recognition. Extensive experiments on five datasets namely, CMU-PIE, Multi-PIE, ChokePoint, HFB and IXMAS datasets and comparisons with several state-of-the-art approaches show the effectiveness of the proposed approach. (C) 2015 Elsevier B.V. All rights reserved.
Resumo:
Peptide metabolism forms an important part of the metabolic network of Salmonella and to acquire these peptides the pathogen possesses a number of peptide transporters. Whilst various peptide transporters known in Salmonella are well studied, very little is known about the carbon starvation (cst) genes cstA and yjiY, which are also predicted to be involved in peptide metabolism. We investigated the role of these genes in the metabolism and pathogenesis of Salmonella, and demonstrated for the first time, to the best of our knowledge, that cst genes actually participate in transport of specific peptides in Salmonella. Furthermore, we established that the carbon starvation gene yjiY affects the expression of flagella, leading to poor adhesion of the bacterium to host cells. In contrast to the previously reported role of cstA in virulence of Salmonella in Caenorhabditis elegans, we showed that yjiY is required for successful colonization of Salmonella in the mouse gut. Thus, cst genes not only contribute to the metabolism of Salmonella, but also influence its virulence.
Resumo:
The biodegradation of polycaprolactone (PCL), polylactic acid (PLA), polyglycolide (PGA) and their copolymers, poly (lactide-co-glycolide) and poly (D, L-lactide-co-caprolactone) (PLCL) was investigated. The influence of different solvents on the degradation of these polymers at 37 degrees C in the presence of two different lipases namely Novozym 435 and the free lipase of porcine pancreas was investigated. The rate coefficients for the polymer degradation and enzyme deactivation were determined using continuous distribution kinetics. Among the homopolymers, the degradation of PGA was nearly an order of magnitude lower than that for PCL and PLA. The overall rate coefficients of the copolymers were higher than their respective homopolymers. Thus, PLCL degraded faster than either PCL or PLA. The degradation was highly dependent on the viscosity of the solvent used with the highest degradation observed in acetone. The degradation of the polymers in acetone was nearly twice that observed in dimethyl sulfoxide indicating that the degradation decreases with increase in the solvent viscosity. The degradation of the polymers in water-solvent mixtures indicated an optimal water content of 2.5 wt% of water.
Resumo:
Ready-to-use screen printed glucose sensors are fabricated using Prussian Blue (PB) and Cobalt Phthalocyanine (CoPC) mediated carbon inks as working electrodes. The reference and counter electrodes are screen printed using silver/silver chloride and graphitic carbon paste respectively. The screen printed reference electrodes (internal reference electrode (IRE)) are found to be stable for more than 60 minutes when examined with saturated calomel electrode. Optimal operating voltage for PB and CoPC screen printed sensors are determined by hydrodynamic voltammetric technique. Glucose oxidase is immobilized on the working electrodes by cross-linking method. PB mediated glucose sensor exhibits a sensitivity of 5.60 mA cm(-2)/mM for the range, 10 to 1000 mu M. Sensitivity of CoPC mediated glucose sensor is found to be 5.224 mu A cm(-2)/mM and amperometeric response is linear for the range, 100 to 1500 mu M. Interference studies on the fabricated glucose sensors are conducted with species like uric acid and ascorbic acid. PB mediated sensors showed a completely interference-free behavior. The sensing characteristics of PB mediated glucose sensors are also studied in diluted human serum samples and the results are compared with the values obtained through standard clinical method. The co-efficient of variation is found to be less than 5%. (C) 2015 The Electrochemical Society. All rights reserved.