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Alternate assemblies of platinum nanoparticles and metalloporphyrins as tunable electrocatalysts for dioxygen reduction.

Through electrostatic layer-by-layer (LBL) assembly, negatively charged citrate-stabilized platinum nanoparticles (PtNPs) and positively charged [tetrakis(N-methylpyridyl)porphyrinato] cobalt were alternately deposited on a 4-aminobenzoic acid-modified glassy carbon electrode and also on indium tin oxide substrates, directly forming the three-dimensional nanostructured materials. Thus-prepared multilayer films were characterized by UV--visible spectroscopy, surface plasmon resonance (SPR) spectroscopy, atomic force microscopy (AFM), and cyclic voltammetry. Regular growth of the multilayer films is monitored by UV--visible spectroscopy and SPR spectroscopy. AFM provides the morphology of the multilayer films. The PtNPs containing multilayer films exhibit high electrocatalytic activity for the reduction of dioxygen with high stability. Rotating disk electrode voltammetry and rotating ring-disk electrode voltammetry demonstrate that the PtNP-containing multilayer films can catalyze an almost four-electron reduction of O(2) to H(2)O in an air-saturated 0.5 M H(2)SO(4) solution. Furthermore, the electrocatalytic activity of the films could be further tailored by simply choosing different cycles in the LBL process or more specifically the amount of the assembly components in the films. The high electrocatalytic activity and good stability for dioxygen reduction make the PtNP-containing multilayer films potential candidates for the efficient cathode material in fuel cells.

Metalloporphyrins↗

Three-dimensional stabilization provided by the external spinal fixator compared to two internal fixation devices: a biomechanical in vitro flexibility study.

We performed an in vitro study to investigate the stabilization (i.e. motion reduction) provided by the external spinal fixator (ESF), and to compare the three configurations of the ESF with two internal fixation techniques. Six human cadaveric lumbar spine specimens (L3-S1) were subjected to multidirectional flexibility testing in six configurations: (1) intact, (2) ESF in neutral, (3) ESF in distraction, (4) ESF in compression, (5) translaminar facet screw fixation, and (6) internal transpedicular fixation. Both the ESF and the internal fixation systems stabilized the specimens from L4 to S1. In each testing configuration, pure bending moments of flexion-extension, bilateral axial rotation, and bilateral lateral bending were applied to the uppermost vertebra stepwise to a maximum of 10 Nm. The rigid body motion between the vertebrae was measured using an optoelectronic camera system, and custom software was used to calculate the intervertebral rotations. For each applied motion in all testing configurations, the total range of motion (ROM) of L4-S1 is reported. All three ESF configurations stabilized the spine significantly when compared to the intact specimen. The ESF in compression provided significantly more stabilization in flexion-extension than the two other ESF configurations, but no other significant differences were found between the three ESF modes. In flexion-extension the ESF stabilized the spine significantly when compared with the two internal fixation devices. Only in bilateral lateral bending was the ESF inferior to internal transpedicular fixation in providing stabilization. The results of the present study suggest that the ESF provides a high degree of stabilization for preoperative assessment of selected low back pain patients. Whether other non-mechanical factors affect the pain relief experienced by the patients remains unknown.

Adult↗

Non-linear reduction for kinetic models of metabolic reaction networks.

Kinetic models of metabolic networks are essential for predicting and optimizing the transient behavior of cells in culture. However, such models are inherently high dimensional and stiff due to the large number of species and reactions involved and to kinetic rate constants of widely different orders of magnitude. In this paper we address the problem of deriving non-stiff, reduced-order non-linear models of the dominant dynamics of metabolic networks with fast and slow reactions. We present a method, based on singular perturbation analysis, which allows the systematic identification of quasi-steady-state conditions for the fast reactions, and the derivation of explicit non-linear models of the slow dynamics independent of the fast reaction rate expressions. The method is successfully applied to detailed models of metabolism in human erythrocytes and Saccharomyces cerevisiae.

Carbon↗

A cis-proline to alanine mutant of E. coli aspartate transcarbamoylase: kinetic studies and three-dimensional crystal structures.

The only cis-proline residue in Escherichia coli aspartate transcarbamoylase has been replaced by alanine using site-specific mutagenesis. The Pro268-->Ala enzyme exhibits a 40-fold reduction in enzyme activity and decreased substrate affinity toward carbamoyl phosphate and aspartate compared to the corresponding values for the wild-type enzyme. The concentration of the bisubstrate analogue N-phosphonacetyl-L-aspartate (PALA) required to activate the mutant enzyme to the same extent as the wild-type enzyme is significantly increased. The heterotropic effects of ATP and CTP upon the Pro268-->Ala enzyme are also altered. Crystal structures of the Pro268-->Ala enzyme in both T- and R-states show that the cis-peptidyl linkage between Leu267 and Ala268 is maintained. However, the tertiary structure of both the catalytic and regulatory chains has been altered by the amino acid substitution, and the mobility of the active-site residues is increased for the R-state structure of Pro268-->Ala enzyme as comparison with the wild-type R-state structure. These structural changes are responsible for the loss of enzyme activity. Thus, Pro268 is required for the proper positioning of catalytically critical residues in the active site and is important for the formation of the high-activity high-affinity R-state of E. coli aspartate transcarbamoylase.

Alanine↗

Caring Behaviors Inventory: a reduction of the 42-item instrument.

BACKGROUND: Caring traditionally has been at the center of nursing. Effectively measuring the process of nurse caring is vital in nursing research. A short, less burdensome dimensional instrument for patients' use is needed for this purpose. OBJECTIVES: To derive and validate a shorter Caring Behaviors Inventory (CBI) within the context of the 42-item CBI. METHODS: The responses to the 42-item CBI from 362 hospitalized patients were used to develop a short form using factor analysis. A test-retest reliability study was conducted by administering the shortened CBI to new samples of patients (n = 64) and nurses (n = 42). RESULTS: Factor analysis yielded a 24-item short form (CBI-24) that (a) covers the four major dimensions assessed by the 42-item CBI, (b) has internal consistency (alpha =.96) and convergent validity (r =.62) similar to the 42-item CBI, (c) reproduces at least 97% of the variance of the 42 items in patients and nurses, (d) provides statistical conclusions similar to the 42-item CBI on scoring for caring behaviors by patients and nurses, (e) has similar sensitivity in detecting between-patient difference in perceptions, (f) obtains good test-retest reliability (r = .88 for patients and r=.82 for nurses), and (g) confirms high internal consistency (alpha >.95) as a stand-alone instrument administered to the new samples. CONCLUSION: CBI-24 appears to be equivalent to the 42-item CBI in psychometric properties, validity, reliability, and scoring for caring behaviors among patients and nurses. These results recommend the use of CBI-24 to reduce response burden and research costs.

Academic Medical Centers↗

Stability and structural analysis of alpha-amylase from the antarctic psychrophile Alteromonas haloplanctis A23.

The alpha-amylase secreted by the antarctic bacterium Alteromonas haloplanctis displays 66% amino acid sequence similarity with porcine pancreatic alpha-amylase. The psychrophilic alpha-amylase is however characterized by a sevenfold higher kcat and kcat/Km values at 4 degrees C and a lower conformational stability estimated as 10 kJ.mol-1 with respect to the porcine enzyme. It is proposed that both properties arise from an increase in molecular flexibility required to compensate for the reduction of reaction rates at low temperatures. This is supported by the fast denaturation rates induced by temperature, urea or guanidinium chloride and by the shift towards low temperatures of the apparent optimal temperature of activity. When compared with the known three-dimensional structure of porcine pancreatic alpha-amylase, homology modelling of the psychrophilic alpha-amylase reveals several features which may be assumed to be responsible for a more flexible, heat-labile conformation: the lack of several surface salt bridges in the (beta/alpha)8 domain, the reduction of the number of weakly polar interactions involving an aromatic side chain, a lower hydrophobicity associated with the increased flexibility index of amino acids forming the hydrophobic clusters and by substitutions of proline for alanine residues in loops connecting secondary structures. The weaker affinity of the enzyme for Ca2+ (Kd = 44 nM) and for Cl- (Kd = 1.2 mM at 4 degrees C) can result from single amino acid substitutions in the Ca(2+)-binding and Cl(-)-binding sites and can also affect the compactness of alpha-amylase.

Amino Acid Sequence↗

The predictive value of mitral leaflet motion and thickness index scores on early restenosis after mitral balloon valvuloplasty.

The purpose of this study was to investigate whether there is any association between mitral leaflet motion (LMI) and leaflet thickness index (LTI) scores and the rate of restenosis 3 months after successful mitral balloon valvuloplasty. The study population consisted of 46 patients with symptomatic rheumatic mitral stenosis who underwent balloon valvuloplasty (37 women, 9 men; mean age, 36 +/- 9 years). Two-dimensional and Doppler echocardiography were performed in all patients on the day before, immediately after, and 3 months after valvuloplasty. The severity of restriction of leaflet motion and the severity of leaflet thickening were classified into grades of mild (a score of 0), moderate (a score of 1), and severe (a score of 2). Subvalvular disease and commissural involvement were homogeneous in all patients. Before and immediately after mitral balloon valvuloplasty, there were no significant differences in mitral valve area among the groups with different LMI and LTI scores. However, at 3 months after valvuloplasty, reduction in mitral valve area was more significant in patients who had higher pre-procedural LMI and LTI scores (P < 0.05). The rates of early restenosis were 0 with a total score of 0, 14.2% with a total score of 1-2, and 32% with a total score of 3. In conclusion, quantitative assessment of LMI and LTI scores by 2-dimensional echocardiography may be helpful in predicting early restenosis after mitral balloon valvuloplasty. Early reduction in mitral valve area is significant in patients who have higher total LMI and LTI scores.

Adolescent↗

Matrix embedding alters the immune response against endothelial cells in vitro and in vivo.

BACKGROUND: Endothelial cell (EC) dysfunction represents the first manifestation of atherosclerotic disease. Restoration of endothelium via seeding or transfection is hampered by local alterations in flow, inflammation, and metabolic activation. Perivascular EC matrix implants are shielded from these forces and still control vascular repair. The host immune response to such implants, however, remains largely unknown. We investigated the effect of embedding of ECs within 3-dimensional matrices on host immune responses in vitro and in vivo. METHODS AND RESULTS: We compared expression of major histocompatibility complex (MHC), costimulatory, and adhesion molecules by free aortic ECs or ECs embedded in Gelfoam matrices by flow-cytometry. T-cell proliferation was assessed by [3H] thymidine incorporation. Humoral immune response (ELISA and FACS analysis) and cellular (histopathology) infiltration were investigated after subcutaneous injection of free porcine aortic ECs (PAEs) or of a Gelfoam/EC block, or after concomitant injection of PAEs adjacent to Gelfoam in rats. Aortic ECs embedded in Gelfoam expressed lower levels of MHC class II, costimulatory, and adhesion molecules compared with free ECs (P<0.001), and induced 3-fold less proliferation of human CD4+ T-cells (P<0.0005). Implantation of a Gelfoam/EC block in rats nearly abrogated the immune response with 1.75- to 9.0-fold downregulation in tumor necrosis factor-alpha, interleukin-6, monocyte chemotactic protein-1, and PAE-specific immunoglobulin G (P<0.005) and 3.3- to 4.5-fold reduction in leukocytic tissue infiltration. Injecting PAEs adjacent to Gelfoam induced a significant response comparable to that of free implanted PAEs. CONCLUSIONS: Embedding ECs within 3-dimensional matrices alters the host immune response by inhibiting expression of MHC class II, costimulatory, and adhesion molecules, offering the rationale to develop novel therapies for vascular diseases.

Animals↗

Hepatic protein pattern alterations following perfluorodecanoic acid exposure in rats.

The effect after 8 days of 20 and 50 mg/kg in vivo exposure to perfluoro-n-decanoic acid (PFDA) on the protein pattern of rat liver whole homogenates was studied using high-resolution, large-scale two-dimensional polyacrylamide gel electrophoresis. PFDA exposure altered 13 proteins reproducibly in each of 5 samples tested. While most of the altered proteins remained unidentified, several known mitochondrial proteins were induced by PFDA as were others in a dose-related manner. Conversely, PFDA caused the reduction of albumin as well as several unknown proteins, also in a dose-related manner. Cytoskeletal proteins were unaffected by PFDA. The present results suggest that PFDA's toxic mechanism may involve more than membrane integration and lipid accumulation. Changes in the two-dimensional protein pattern indicate that selective protein synthetic or catabolic processes may undergo qualitative alterations after PFDA treatment as well.

Animals↗

Experiments with the nonlinear and chaotic behaviour of the multiplicative algebraic reconstruction technique (MART) algorithm for computed tomography.

Among the iterative reconstruction algorithms for tomography, the multiplicative algebraic reconstruction technique (MART) has two advantages that make it stand out from other algorithms: it confines the image (and therefore the projection data) to the convex hull of the patient, and it maximizes entropy. In this paper, we have undertaken a series of experiments to determine the importance of MART nonlinearity to image quality. Variants of MART were implemented aiming to exploit and exaggerate the nonlinear properties of the algorithm. We introduce the Power MART, Boxcar Averaging MART and Bouncing MART algorithms. Power MART is linked to the relaxation concept. Its behaviour is similar to that of the chaos of a logistic equation. There appears to be an antagonism between increasing nonlinearity and noise in the projection data. The experiments confirm our general observation that regularization as a means of solving simultaneous linear equations that are underdetermined is suboptimal: it does not necessarily select the correct image from the hyperplane of solutions, and so does not maximize the image quality:x-ray dose ratio. Our investigations prove that there is scope to optimize CT algorithms and thereby achieve greater dose reduction.

Algorithms↗

3-Dimensional rotational X-ray imaging, 3D-RX: image quality and patient dose simulation for optimisation studies.

We have developed a full-scale simulation model as a tool for X-ray system design, image quality optimisation and patient dose reduction. The model supports the (de-)composition of system level requirements and takes inherent care of the prerequisite mutual coherence between component requirements. Furthermore, the model is equipped with several types of automatic X-ray control loop simulations. All relevant patient dose and image quality items are calculated, such as contrast, sharpness, lag and noise and finally combined in 'IQ figures of merit'. The short calculation times realised enable comprehensive multi-parameter optimisation studies. Initially, the IQ model dealt with the performance of 'flat' projection X-ray systems referred to as two-dimensional (2D). Nowadays, 2D X-ray systems are increasingly used for three-dimensional rotational X-ray imaging (3D-RX). An additional 3D-RX module, dealing with patient dose as well as image quality, will simplify and accelerate specific 3D-RX optimisations concerning, e.g. exposure factors, dose control concepts and image processing algorithms.

Algorithms↗

Rapid modulation of GABA in sensorimotor cortex induced by acute deafferentation.

Recovery of function after acute injury to the central nervous system may be controlled by the availability of gamma-aminobutyric acid (GABA), the main inhibitory neurotransmitter in the cerebral cortex. Acute lesions as well as manipulation of sensory inputs can lead to rapid reorganization of the cerebral cortex, occurring within minutes to hours. Reduction of cortical inhibitory tone through a decrease in the availability of GABA has been suggested as a possible mechanism; however, the degree and temporal course of the changes in brain GABA are not known. A novel method using two-dimensional J-resolved magnetic resonance spectroscopy showed that GABA levels in the human sensorimotor cortex are quickly reduced within minutes of deafferentation. This finding strongly supports the view that the release of latent corticocortical projections from tonic inhibition through decreased GABA availability is a mechanism of rapid cortical plasticity. Reduction of brain GABA can play a pivotal role in regulating the extent of rapid cortical reorganization after lesions or changes in sensory input.

Adult↗

A structural transition in d(AT)n.d(AT)n inserts within superhelical DNA.

We have constructed plasmids carrying d(AT)n.d(AT)n inserts of different lengths. Two-dimensional gel electrophoresis patterns show that an increase in the negative superhelicity of these DNAs brings about a structural transition within the inserts, resulting in a reduction of the superhelical stress. However, this reduction corresponds to the expected values neither for cruciform nor the Z form. Those DNA topoisomers in which the structural transition had occurred proved to be specifically recognizable by single-strand-specific endonuclease S1, with the cleavage site situated at the centre of the insert. These data, as well as kinetic studies, suggest that the cloned d(AT)n.d(AT)n sequences adopt a cruciform rather than the Z-form structure. We discuss plausible reasons of the discrepancy between the observed superhelical stress release and that expected for the transition of the insert to the cruciform state.

Base Sequence↗

In-situ oxidation of trichloroethene by permanganate: effects on porous medium hydraulic properties.

In-situ oxidation of dense nonaqueous-phase liquids (DNAPLs) by strong oxidants such as potassium permanganate (KMnO4) has been proposed as a possible DNAPL remediation strategy. In this study, we investigated the effects of in-situ trichloroethene (TCE) oxidation by KMnO4 on porous medium hydraulic properties. In particular, we wanted to determine the overall effects of concurrent solid phase (MnO2) precipitation, gas (CO2) evolution and TCE dissolution resulting from the oxidation reaction on the porous medium's aqueous-phase relative permeability, krw. Three TCE removal experiments were conducted in a 95-cm long, 5.1-cm i.d. glass column, which was homogeneously packed with well-characterized 30/40-mesh silica sand. TCE was emplaced in the sand-pack in residual, entrapped form through a sequence of water/TCE imbibition and drainage steps. The column was then flushed under constant aqueous flux conditions for up to 104 h with either deionized water (reference experiment), deionized water containing 5 mM KMnO4 or deionized water containing 5 mM KMnO4 and 300 mM Na2HPO4. Aqueous-phase relative permeabilities were computed from measured flow rates and measurements of aqueous-phase pressure head, h obtained using pressure transducers connected to tensiometers distributed along the column length. A dual-energy gamma radiation system was used to monitor changes in fluid saturation that occurred during each experiment. In addition, column effluent samples were collected for chemical analyses. Dissolution of TCE during deionized water flushing led to an increase in krw by approximately 22% and a local reduction in h. On the other hand, vigorous CO2 gas production and precipitation of MnO2 was visually observed during flushing with deionized water that contained 5 mM KMnO4. As a consequence, krw declined by approximately 96% and h increased locally by more than 1000 cm H2O during the first 24 h of the experiment, causing sand-pack ruptures and pump failure. Conversely, less CO2 gas production and MnO2 precipitation was visually observed during flushing with deionized water that contained 5 mM KMnO4 and 300 mM Na2HPO4. Consequently, only small increases in h (< 15 cm H2O) were observed in this experiment due to a reduction in krw of approximately 53%. While we must attribute changes in h due to variations in krw to our specific experimental design (constant aqueous flux, one-dimensional flow experiments), these experiments nevertheless confirm that successful application of in situ chemical oxidation of TCE requires consideration of detrimental processes such as MnO2 precipitation and CO2 gas formation. In addition, our results indicate that utilization of a buffered oxidant solution may improve the effectiveness of in-situ oxidation of TCE by KMnO4 in otherwise weakly buffered porous media.

Chemical Precipitation↗

[Personal experience with treatment of acetabular fractures].

PURPOSE OF THE STUDY: The article presents a retrospective evaluation of a group of 119 patients treated for acetabular fractures between 1996 and 2002. MATERIAL: In the monitored period 119 patients, (89 men, 30 women) average age 35 years, were hospitalized with an acetabular fracture. Surgically treated were 91 patients (70 men, 21 women), average age 29 years. Prevailing in this group were Type A fractures which accounted for 45%, Type B was represented by 35% and Type C included 19%. Twenty-eight patients were treated conservatively. METHOD: Indication for surgical treatment was instability, hip incongruence or combination of both. Conservative treatment was chosen in the fractures without displacement, in fractures with secondary congruency and in case of contraindication of surgical treatment. The indication was always based on CT examination with a three-dimensional reconstruction. The surgery was performed within 24 hours in 42%, within 48 hours in 64%, within 72 hours in 87%. Posterior Kocher-Langenbeck approach was used in 54%, anterior ilioinguinal approach in 22% and the combination of both approaches in 24%. Posterior approach was indicated in A1, A2, B1 fractures, ilioinguinal approach in A3, B3, C1 Types, the combination of both approaches in B2, C2, C3 fractures. Osteosynthetic material was chosen according to the fracture type, bone quality and fragment size. RESULTS: The evaluation was based on subjective complaints, radiograph and clinical examination using Harris Hip Score. In the group of 91 patients treated surgically we achieved excellent result in 41 cases (45%), very good result in 28 cases (31%), fair result in 7 cases (9%) and poor result in 13 cases (15%). We recorded 17 peroperative complications--11 times inadequate reduction, 5 times nerve injury, once vascular injury. Early complications included infect--twice and loosening of implant--once. Late complications occurred in 26 cases--paraarticular ossification in 17 cases, aseptic necrosis of the femoral head in 8 cases. DISCUSSION: The treatment of fractures of the acetabulum as any intraarticular fracture is based on a precise anatomical reduction, stable fixation and early mobilization. Difficulties of the treatment of the acetabulum arise from the anatomical relations. Decisive for the indication, the method of treatment and prognosis is the type of the injury of the weight-bearing area of the acetabulum. In order to understand the type of the fracture it is necessary to make a careful preoperative examination based particularly on CT examination with a three-dimensional reconstruction. The indication and treatment strategy--surgical approach, method of reduction, type of fixation--is selected accordingly. Each surgical approach has its limits and also the possibilities of fixation are limited by the type of the fracture. The outcomes depend directly on the quality of the reconstruction of the articular surface and the incidence of complications, caused either by the injury or preoperatively. CONCLUSION: The treatment of displaced fractures of the acetabulum is on principle surgical. Satisfactory results are produced only by anatomical reconstruction of the weight-bearing area of the acetabulum. Surgical treatment is highly demanding and the long-term results not always correspond to the efforts made.

Acetabulum↗

Depression of progenitor cell function by advanced glycation endproducts (AGEs): potential relevance for impaired angiogenesis in advanced age and diabetes.

Diabetes and ageing induce reduction and dysfunction of vascular progenitor cells. Advanced glycation endproducts (AGEs) accumulate in diabetes and ageing. We investigated the influence of AGEs on function of CD34 progenitor cells. CD34 cells were co-cultured with HUVECs in a three-dimensional spheroid assay. Sprout length growth and incorporation of CD34 cells into the sprouts were analyzed under 2, 20 or 200 microg/ml AGEs. AGE-receptor expression, MAP-kinase signal transduction and apoptosis were analyzed using PCR, Western blotting and flow cytometry. In the spheroid assay, AGEs concentration-dependently cause a reduction of sprout length growth by 6+/-6 to 32+/-6% and an attenuation of progenitor cells incorporation into the sprouting endothelium by up to 43+/-6%. This functional impairment is accompanied by activation of CD34 cell proliferation at lower concentrations (2 or 20 microg/ml) and by apoptosis activation under 200 microg/ml AGEs. The mRNA expression of the receptors for AGEs and the AGEs-induced activation of p38 and p44/42 MAP-kinases are demonstrable in CD34 cells. This AGEs-mediated impairment of progenitor cell function identifies a new pathophysiological mechanism of disturbed vascular adaptation in diabetes or ageing and suggests that lowering AGEs in recipients of progenitor cell therapy might be beneficial for the success of this therapy.

Aging↗

Dissimilar effect of perming and bleaching treatments on cuticles: advanced hair damage model based on elution and oxidation of S100A3 protein.

Hair treatment chemicals induce sudden and severe hair damage. In this study, we examined cuticles from untreated, permed, and bleached hair that were mechanically discriminated by shaking in water. Both perming and bleaching treatments are prone to easily delaminate cuticles. Confocal microscopy revealed that the cuticles of permed hair were delaminated with larger pieces than untreated ones. On the other hand, the cuticles of bleached hair tend to fragment into small peptides. At the minimum concentration of thioglycolate required to elute S100A3 protein from the endocuticle into the reductive permanent waving lotion, enlarged delaminated cuticle fragments were observed. Although S100A3 is retained in bleached hair, S100A3 is irreversibly oxidized upon bleaching treatment. It is likely that the oxidative cleavage of disulfide bonds between cuticle-constituting proteins, including S100A3, results in the fragile property of cuticles. Here we present a more comprehensive model of hair damage based on a diverse mechanism of cuticle delamination.

Electrophoresis, Gel, Two-Dimensional↗