Site-selected mutagenesis of a conserved nucleotide binding HXGH motif located in the ATP sulfurylase domain of human bifunctional 3'-phosphoadenosine 5'-phosphosulfate synthase

3'-Phosphoadenosine-5'-phosphosulfate (PAPS) synthase is a bifunctional protein consisting of an NH2-terminal APS kinase and a COOH-terminal ATP sulfurylase. Both catalytic activities require ATP; the APS kinase domain involves cleavage of the beta-gamma phosphodiester bond of ATP, whereas the ATP sulfurylase domain involves cleavage of the alpha-beta phosphodiester bond of ATP. Previous mutational studies have suggested that beta-gamma phosphodiesterase activity involves a highly conserved NTP-binding P-loop motif located in the adenosine-5'-phosphosulfate kinase domain of PAPS synthases. Sequence alignment analysis of PAPS synthases and the superfamily of TagD-related nucleotidylyltransferases revealed the presence of a highly conserved HXGH motif in the ATP sulfurylase domain of PAPS synthases, a motif implicated in the alpha-beta phosphodiesterase activity of cytidylyltransferases. Thus, site-selected mutagenesis of the HXGH motif in the ATP sulfurylase domain of human PAPS synthase (amino acids 425-428) was performed to examine this possibility. Either H425A or H428A mutation produced an inactive enzyme. In contrast, a N426K mutation resulted in increased enzymatic activity. A G427A single mutant resulted in only a modest 30% reduction in catalytic activity, whereas a G427A/H428A double mutant produced an inactive enzyme. These results suggest an important role for the HXGH histidines in the ATP sulfurylase activity of bifunctional PAPS synthase and support the hypothesis that the highly conserved HXGH motif found in the ATP sulfurylase domain of PAPS synthases is involved in ATP binding and alpha-beta phosphodiesterase activity.     

Synthesis of novel optically active pyrrolidine‐containing polyaniline: A new heterogeneous organo polymeric‐base catalyst for direct Aldol reaction

An optically active pyrrolidine containing polyaniline, poly‐(2‐anilinomethylpyrrolidine) (PANiMp) is synthesized for the first time. PANiMp material is synthesized by simple one‐pot synthetic route and is characterized by circular dichroism, NMR, FTIR, and electronic absoroption spectral techniques. This novel polyaniline base is evaluated as new heterogeneous organo polymeric‐base catalyst for direct Aldol reaction in water medium. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010     

Resource settings have a major influence on the outcome of maintenance hemodialysis patients in South India

Chronic kidney disease is reaching epidemic proportions and the number of patients on renal replacement therapy (RRT) is increasing worldwide and also in developing countries. To meet the challenge of providing RRT, a few charity organizations provide hemodialysis units for underprivileged patients, as the private hospitals are unaffordable for the majority. There is a paucity of information on the outcome of dialysis in these patients. Here, we describe the outcome of hemodialysis patients comparing the middle‐ and upper‐class income group with the lower class income group. A retrospective analysis was carried out in 558 CKD patients initiated on maintenance hemodialysis in two different dialysis facilities. Group A (n=247) included those who belonged to the lowermost socioeconomic status and were undergoing dialysis in two nonprofit, charity (TANKER)‐run dialysis units, and Group B (n=311) was undergoing dialysis in a nonprofit hospital setting where no subsidy was given. Those patients of a low socioeconomic status, especially those who are diabetics, have a higher death rate (Group A‐38.1%, Group B‐4.2%) and loss to follow‐up (Group A‐25.9%, Group B‐0.3%) compared with those who are in the middle‐ and high‐income group. Higher EPO use and hence higher hemoglobin levels (Group A‐6.4±1.2, Group B‐8.9±1.5 P<0.001) were observed in those who were in the middle and the higher income group. Lower serum phosphorus level was observed in the low‐socioeconomic group (Group A‐4.7±1.5, Group B‐5.5±1.9, P<0.001). Patients belonging to the middle and higher socioeconomic group undergo more transplantations compared with the lower socioeconomic group (Group A‐2.4%, Group B‐65.6%).     

Effect of the solvent on growth of titania nanotubes prepared by anodization of Ti in HCl

The effect of the solvent on the anodic growth of titania nanotubes in HCl dissolved in water, ethylene glycol and 2-propanol was studied. These nanotubes grow with locally rapid breakdown of the passive TiO₂ film forming a forest of nanotubes-bearing microtowers with the background of passive TiO₂ Film. These bundles of assembled-groups of titania nanotubes look like Pillars corals. The low relative permittivity of the 2-propanol led to lowering of dissociation of HCl and hence lowering the activity of H⁺ and Cl⁻ ions which in turn led to suppress of dissolution of titania and increasing the growth rate of the titania nanotubes. The X-ray diffraction pattern showed that the titania nanotubes after annealing change to the crystalline anatase phase. The anodic films showed characteristic coloration with intensity and color that changes (qualitatively) with time of anodization.     

In vitro and in vivo investigations of upconversion and NIR emitting Gd2O3:Er3+,Yb3+ nanostructures for biomedical applications

The use of an “over 1000-nm near-infrared (NIR) in vivo fluorescence bioimaging” system based on lanthanide containing inorganic nanostructures emitting in the visible and NIR range under 980-nm excitation is proposed. It may overcome problems of currently used biomarkers including color fading, phototoxicity and scattering. Gd₂O₃:Er³⁺,Yb³⁺ nanoparticles and nanorods showing upconversion and NIR emission are synthesized and their cytotoxic behavior is investigated by incubation with B-cell hybridomas and macrophages. Surface modification with PEG-b-PAAc provides the necessary chemical durability reducing the release of toxic Gd³⁺ ions. NIR fluorescence microscopy is used to investigate the suitability of the nanostructures as NIR–NIR biomarkers. The in vitro uptake of bare and modified nanostructures by macrophages is investigated by confocal laser scanning microscopy. In vivo investigations revealed nanostructures in liver, lung, kidneys and spleen a few hours after injection into mice, while most of the nanostructures have been removed from the body after 24?h.     

Isopropylation of ethylbenzene over MCM-22 molecular sieve

MCM-22 materials (Si/Al ratios 24, 50 and 75) were synthesized and characterized. The catalytic activity was examined in the vapour phase isopropylation of ethylbenzene with isopropyl alcohol. Based on ethylbenzene conversion, the order of activity of the catalysts is found to be MCM-22(50) > MCM-22(24) = MCM-22(75). The selective formation of p-isopropyl ethylbenzene (p-IPEB) suggests that the reaction occurs mainly inside the 10-membered ring channel. The time on stream study over MCM-22(50) showed steady conversion for 6 h with nearly the same selectivity to p-isopropyl ethylbenzene (p-IPEB) and o-isopropyl ethylbenzene (p-IPEB).     

Effect of baffle–cavity ratios on buoyancy convection in a cavity with mutually orthogonal heated baffles

Buoyancy driven convection in a square cavity induced by two mutually orthogonal and arbitrarily located baffles is studied numerically. The baffles are of different sizes and the flow is two-dimensional. The coupled governing equations were solved by finite difference method using Alternating Direction Implicit technique and Successive Over-Relaxation method. The steady state results are presented in the form of streamline and isotherm plots. It is found that buoyancy force plays a key role and overall heat transfer in the cavity is enhanced for higher values of both baffle–cavity ratios. Flow inhibition emerges as a deciding factor and diminishes heat transfer when the horizontal baffle is located above the center of the cavity.     

Biopolishing of cotton fabrics with total cellulases of Trichoderma reesei and optimization using Taguchi methods

Biopolishing of cotton fabrics enhances appearance and handle of the fabrics without compromising on essential properties. Process of biopolishing is influenced by concentration of cellulases, temperature, pH, and duration of treatment, besides the activity levels of enzymes, method of mechanical agitations and construction features of fabrics. Optimization of process parameters, including mechanical agitations and fabric construction features, has been carried out using Taguchi methods followed by analysis of variance and confirmation tests. All the design parameters, used in the study, have predominant influence on weight loss, fabric strength after biopolishing while thickness, bursting strength, abrasion losses, and flexural rigidity of the fabrics were significantly influenced by the concentration of cellulases together with duration of treatment. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009