Glutathione immunosensing platform based on total internal reflection ellipsometry enhanced by functionalized gold nanoparticles.

An immunosensor to detect small molecules, such as glutathione (GSH), has been developed by combination of ellipsometry and Kretschmann surface plasmon resonance (SPR). The Au thin film used for surface plasmon polariton (SPP) excitation is functionalized with anti-GSH to specifically bind GSH. At low concentrations, the small refractive index changes caused by the low molecular weight of GSH induced only negligible shifts in the plasmon resonant energy during GSH binding. To improve sensitivity, gold nanoparticles (AuNPs) are functionalized with glutathione acting as amplifiers of the antigen-antibody interaction. Changes induced by the AuNP adsorption are monitored using Ψ and Δ ellipsometric functions. After performing competitive assays using solutions containing different concentrations of free GSH and a constant amount of functionalized AuNPs, it was concluded that the resonant energy linearly shifts as the relative concentration of free GSH increases. A detection limit for free GSH in the nanomolar range is found, demonstrating the effectiveness of AuNPs to enhance the sensitivity to immunoreactions in total internal reflection ellipsometry.

Electrochemically generated nanoparticles of halogen-bridged mixed-valence binuclear metal complex chains.

Spherical nanoparticles composed of MMX chains can be made by a polymerization strategy driven by electrochemical processes. In particular, the [Pt2(MeCS2)4I2] (MMI2) dimetal subunit is employed as a monomer for the formation of [Pt2(MeCS2)4I]n spherical nanostructures on surfaces. We have paid particular attention to elucidating the general mechanism of the deposition process on the basis of in situ electrochemical measurements. The reduction of MMI2 to give the electrodeposition of nanostructures agrees well with formation of the reduced [MMI2](-) species followed by a disproportionation mechanism mediated by iodide anions. The chemical composition of the particles was determined by energy-dispersive X-ray spectroscopy (EDX) and X-ray photoelectron spectroscopy (XPS) to reveal the MMI2 polymer.

Renal replacement therapy for autosomal dominant polycystic kidney disease (ADPKD) in Europe: prevalence and survival--an analysis of data from the ERA-EDTA Registry.

BACKGROUND: Autosomal dominant polycystic kidney disease (ADPKD) is the fourth most common renal disease requiring renal replacement therapy (RRT). Still, there are few epidemiological data on the prevalence of, and survival on RRT for ADPKD. METHODS: This study used data from the ERA-EDTA Registry on RRT prevalence and survival on RRT in 12 European countries with 208 million inhabitants. We studied four 5-year periods (1991-2010). Survival analysis was performed by the Kaplan-Meier method and by Cox proportional hazards regression. RESULTS: From the first to the last study period, the prevalence of RRT for ADPKD increased from 56.8 to 91.1 per million population (pmp). The percentage of prevalent RRT patients with ADPKD remained fairly stable at 9.8%. Two-year survival of ADPKD patients on RRT (adjusted for age, sex and country) increased significantly from 89.0 to 92.8%, and was higher than for non-ADPKD subjects. Improved survival was noted for all RRT modalities: haemodialysis [adjusted hazard ratio for mortality during the last versus first time period 0.75 (95% confidence interval 0.61-0.91), peritoneal dialysis 0.55 (0.38-0.80) and transplantation 0.52 (0.32-0.74)]. Cardiovascular mortality as a proportion of total mortality on RRT decreased more in ADPKD patients (from 53 to 29%), than in non-ADPKD patients (from 44 to 35%). Of note, the incidence rate of RRT for ADPKD remained relatively stable at 7.6 versus 8.3 pmp from the first to the last study period, which will be discussed in detail in a separate study. CONCLUSIONS: In ADPKD patients on RRT, survival has improved markedly, especially due to a decrease in cardiovascular mortality. This has led to a considerable increase in the number of ADPKD patients being treated with RRT.

Single-Electron Charging of Thioctic Acid Monolayer-Protected Gold Clusters.

There is great interest in the use of Monolayer-Protected Gold Clusters (AuMPCs) as nanoscale capacitors in aqueous media for nanobiotechnological applications, such as bioelectrocatalysts, biofuel cells, and biosensors. However, AuMPCs exhibiting subattofarad double-layer capacitance at room temperature, and the resolution of single-electron charging, has been mainly obtained in an organic medium with nonfunctional capping ligands. We report here the synthesis of Thioctic Acid Monolayer-Protected Au Clusters (TA-AuMPCs) showing electrochemical single electron quantized capacitance charging in organic and aqueous solutions and when immobilized onto different self-assembled monolayer-modified gold electrodes. The presence of functional carboxylic groups opens a simple strategy for interfacing a nanoparticle assembly to biomolecules for their use as electron donors or acceptors in biological electron transfer reactions.

Thermal energy for the arthroscopic treatment of tears of the triangular fibrocartilage of the wrist.

We evaluated the clinical and occupational outcomes of arthroscopic treatment with electrothermal shrinkage for triangular fibrocartilage complex (TFCC) tears. We retrospectively reviewed 162 patients. All patients had ulnar-sided wrist pain that limited their occupational and sporting activities. The surgical technique consisted of electrothermal collagen shrinkage of the TFCC. Pain relief, range of motion, complications, reoperation rate, time to return to work and workers' compensation claims were evaluated. Exclusion criteria were distal radioulnar joint instability and association of other wrist lesions. Complete pain relief was noted in 80.3% of the patients, incomplete pain relief in 14.8%, and only 4.9% required reoperation because of pain-persistence. The average range of motion was over 90% compared to the opposite hand. Worker's compensation claims were introduced by 20 patients, of which 6 did not return to their previous occupation. Electrodiathermy may be a useful option for arthroscopic treatment of TFCC tears in cases without distal radioulnar joint instability.

Design and Development of Antibody Functionalized Gold Nanoparticles for Biomedical Applications.

Antibody-functionalized gold nanoparticle constitutes a powerful interface biosystem for biomedical applications where the properties of gold nanoparticles and the specificity of antibody-antigen interactions are combined. This study provides insight into the key factors for the development of antibody functionalized gold nanoparticles focusing on the immobilization of the antibody. Here, we address an oriented antibody immobilization procedure on gold nanoparticles. It comprises chelatemodified gold nanoparticles that are designed for oriented immobilization of IgG antibodies (end on spatial orientation) through the metal-chelation to histidine-rich metal binding site in the heavy chain (Fc) of the antibody.

Engineering conductive protein films through nanoscale self-assembly and gold nanoparticles doping.

Protein-based materials are usually considered as insulators, although conductivity has been recently shown in proteins. This fact opens the door to develop new biocompatible conductive materials. While there are emerging efforts in this area, there is an open challenge related to the limited conductivity of protein-based systems. This work shows a novel approach to tune the charge transport properties of protein-based materials by using electron-dense AuNPs. Two strategies are combined in a unique way to generate the conductive solid films: (1) the controlled self-assembly of a protein building block; (2) the templating of AuNPs by the engineered building block. This bottom-up approach allows controlling the structure of the films and the distribution of the AuNPs within, leading to enhanced conductivity. This work illustrates a promising strategy for the development of effective hybrid protein-based bioelectrical materials.

Immobilization of Proteins on Gold Surfaces.

Gold has been a widely used support for protein immobilization in a nonspecific way through electrostatic and hydrophobic interactions. As no tools are available to predict the binding of proteins of biological interest to gold supports-for either nano, micro, or macroscopic sizes-smart, reliable, and reproducible protein immobilization protocols on gold are sought. This chapter will focus on a synthetic strategy which allows for the development of a multiplicity of architectures on gold that may be used for protein immobilization. Because of its simplicity, both from a conceptual and a practical point of view, the strategy demonstrated by this step-by-step synthesis of a functionally self-assembled monolayer (SAM) of thiols on gold is accessible to most laboratories working on enzyme technology, even those with limited organic synthesis facilities.

Correlation and regression analysis between residual gradation and uncorrected visual acuity one year after refractive surgery with LASIK, FS-LASIK, PRK, PRK Xtra techniques and the implantation of ICL® posterior chamber phakic lens in myopic correction.

PURPOSE: To analyze the influence of the final spherical equivalent (SE) in LogMAR uncorrected distance visual acuity (UDVA) one year after refractive surgery. We analysed refractive results, their predictability and efficacy, and the safety results of the different methods as secondary outcomes. SETTING: Refractive Surgery Unit of the Institut Català de Retina (ICR) in Barcelona, Spain. DESIGN: Retrospective, analytical observational study. METHODS: Retrospective and observational study of 654 eyes of 327 patients who underwent refractive surgery to treat their myopia or myopic astigmatism using LASIK, FS-LASIK, PRK, PRK Xtra or ICL-type lens implantation surgery were included. RESULTS: The correlation between the SE in absolute value was statistically significant in all techniques utilized, reaching higher values in the FS-LASIK and LASIK techniques, 0.774 and 0.706 respectively, and lesser values in PRK (0.480) and PRK Xtra (0.482). A significant adjustment via a univariate linear regression model could be implemented in all techniques, albeit the R2 coefficient of determination values were higher than those for the FS-LASIK (0.599) and LASIK (0.494) techniques. CONCLUSIONS: There is a positive correlation between post-surgical SE value and post-operative LogMAR UDVA. These regression models can be adjusted to predict the final UDVA according to the final SE. The techniques that are most influenced by the final SE in terms of their visual results are FS-LASIK and LASIK.

Direct electron transfer to a metalloenzyme redox center coordinated to a monolayer-protected cluster.

A strategy for establishing electrical contact to the metal center of a redox metalloenzyme, galactose oxidase (GOase), by coordination of a linker attached to a monolayer-protected gold cluster is presented. The cluster-enzyme hybrid system was first prepared in solution and characterized by high-angle annular dark-field scanning transmission electron microscopy. Electrochemical communication between a gold electrode and GOase was achieved by first modifying the electrode surface with a biphenyl dithiol self-assembled monolayer followed by reaction with gold clusters capped with thioctic acid. GOase was then immobilized by replacement of the H(2)O molecule at the Cu(II) exogenous site by coordination of a carboxylate-terminated gold cluster. This chemical attachment ensured electrical contact between the redox center and the electrode, leading to direct mediatorless electron transfer to the protein. Hybrid systems can find applications in biosensors and biofuel cells and for studying electrochemically the catalytic mechanism of reactions for which free radicals and electron-transfer reactions are involved. The present results can be extended to other metalloenzymes.

Higher Fecal Short-Chain Fatty Acid Levels Are Associated with Gut Microbiome Dysbiosis, Obesity, Hypertension and Cardiometabolic Disease Risk Factors.

Fiber fermentation by gut microbiota yields short-chain fatty acids (SCFAs) that are either absorbed by the gut or excreted in feces. Studies are conflicting as to whether SCFAs are beneficial or detrimental to cardiometabolic health, and how gut microbiota associated with SCFAs is unclear. In this study of 441 community-dwelling adults, we examined associations of fecal SCFAs, gut microbiota diversity and composition, gut permeability, and cardiometabolic outcomes, including obesity and hypertension. We assessed fecal microbiota by 16S rRNA gene sequencing, and SCFA concentrations by gas chromatography/mass spectrometry. Fecal SCFA concentrations were inversely associated with microbiota diversity, and 70 unique microbial taxa were differentially associated with at least one SCFA (acetate, butyrate or propionate). Higher SCFA concentrations were associated with a measure of gut permeability, markers of metabolic dysregulation, obesity and hypertension. Microbial diversity showed association with these outcomes in the opposite direction. Associations were significant after adjusting for measured confounders. In conclusion, higher SCFA excretion was associated with evidence of gut dysbiosis, gut permeability, excess adiposity, and cardiometabolic risk factors. Studies assessing both fecal and circulating SCFAs are needed to test the hypothesis that the association of higher fecal SCFAs with obesity and cardiometabolic dysregulation is due to less efficient SCFA absorption.

Gut microbiota is associated with obesity and cardiometabolic disease in a population in the midst of Westernization.

Westernization and its accompanying epidemiological transitions are associated with changes in gut microbiota. While the extremes of this lifestyle spectrum have been compared (hunter-gatherers, industrialized countries), populations undergoing such shifts have received little attention. To fill the gap of knowledge about the microbiome evolution following broad lifestyle changes and the emergence of disease-associated dysbiosis, we performed a cross-sectional study in which we characterized the microbiota of 441 Colombian adults through 16S rRNA gene sequencing and determined its relationship with demographic, health-related and dietary parameters. We showed that in the gut microbiota of this cohort thrive taxa proper of both hunter-gatherers (Prevotella, Treponema) and citizens of industrialized countries (Bacteroides, Bifidobacterium, Barnesiella); the relative abundances of these taxa differed from those in Western and non-Western populations. We also showed that the Colombian gut microbiota is composed of five consortia of co-abundant microorganisms that are differentially associated with lifestyle, obesity and cardiometabolic disease, and highlighted metabolic pathways that might explain associations between microbiota and host health. Our results give insights into the evolution of the gut microbiota, and underscore the importance of this community to human health. Promoting the growth of specific microbial consortia could help ameliorating physiological conditions associated with Western lifestyles.

Metformin Is Associated With Higher Relative Abundance of Mucin-Degrading Akkermansia muciniphila and Several Short-Chain Fatty Acid-Producing Microbiota in the Gut.

OBJECTIVE: Recent studies suggest the beneficial effects of metformin on glucose metabolism may be microbially mediated. We examined the association of type 2 diabetes, metformin, and gut microbiota in community-dwelling Colombian adults. On the basis of previous research, we hypothesized that metformin is associated with higher levels of short-chain fatty acid (SCFA)-producing and mucin-degrading microbiota. RESEARCH DESIGN AND METHODS: Participants were selected from a larger cohort of 459 participants. The present analyses focus on the 28 participants diagnosed with diabetes-14 taking metformin- and the 84 participants without diabetes who were matched (3-to-1) to participants with diabetes by sex, age, and BMI. We measured demographic information, anthropometry, and blood biochemical parameters and collected fecal samples from which we performed 16S rRNA gene sequencing to analyze the composition and structure of the gut microbiota. RESULTS: We found an association between diabetes and gut microbiota that was modified by metformin use. Compared with participants without diabetes, participants with diabetes taking metformin had higher relative abundance of Akkermansia muciniphila, a microbiota known for mucin degradation, and several gut microbiota known for production of SCFAs, including Butyrivibrio, Bifidobacterium bifidum, Megasphaera, and an operational taxonomic unit of Prevotella. In contrast, compared with participants without diabetes, participants with diabetes not taking metformin had higher relative abundance of Clostridiaceae 02d06 and a distinct operational taxonomic unit of Prevotella and a lower abundance of Enterococcus casseliflavus. CONCLUSIONS: Our results support the hypothesis that metformin shifts gut microbiota composition through the enrichment of mucin-degrading A. muciniphila as well as several SCFA-producing microbiota. Future studies are needed to determine if these shifts mediate metformin's glycemic and anti-inflammatory properties.

Assembly of designed protein scaffolds into monolayers for nanoparticle patterning.

The controlled assembly of building blocks to achieve new nanostructured materials with defined properties at different length scales through rational design is the basis and future of bottom-up nanofabrication. This work describes the assembly of the idealized protein building block, the consensus tetratricopeptide repeat (CTPR), into monolayers by oriented immobilization of the blocks. The selectivity of thiol-gold interaction for an oriented immobilization has been verified by comparing a non-thiolated protein building block. The physical properties of the CTPR protein thin biomolecular films including topography, thickness, and viscoelasticity, are characterized. Finally, the ability of these scaffolds to act as templates for inorganic nanostructures has been demonstrated by the formation of well-packed gold nanoparticles (GNPs) monolayer patterned by the CTPR monolayer.

Calculating the Rate of Senescence From Mortality Data: An Analysis of Data From the ERA-EDTA Registry.

The rate of senescence can be inferred from the acceleration by which mortality rates increase over age. Such a senescence rate is generally estimated from parameters of a mathematical model fitted to these mortality rates. However, such models have limitations and underlying assumptions. Notably, they do not fit mortality rates at young and old ages. Therefore, we developed a method to calculate senescence rates from the acceleration of mortality directly without modeling the mortality rates. We applied the different methods to age group-specific mortality data from the European Renal Association-European Dialysis and Transplant Association Registry, including patients with end-stage renal disease on dialysis, who are known to suffer from increased senescence rates (n = 302,455), and patients with a functioning kidney transplant (n = 74,490). From age 20 to 70, senescence rates were comparable when calculated with or without a model. However, when using non-modeled mortality rates, senescence rates were yielded at young and old ages that remained concealed when using modeled mortality rates. At young ages senescence rates were negative, while senescence rates declined at old ages. In conclusion, the rate of senescence can be calculated directly from non-modeled mortality rates, overcoming the disadvantages of an indirect estimation based on modeled mortality rates.

Scaffold electrodes based on thioctic acid-capped gold nanoparticles coordinated Alcohol Dehydrogenase and Azure A films for high performance biosensor.

Nanometric size gold nanoparticles capped with thiotic acid are used to coordinate with the Zn (II) present in the catalytic center of Alcohol Dehydrogenase (ADH). In combination with the NADH oxidation molecular catalyst Azure A, electrografted onto carbon screen-printed electrodes, they are used as scaffold electrodes for the construction of a very efficient ethanol biosensor. The final biosensing device exhibits a highly efficient ethanol oxidation with low overpotential of -0.25 V besides a very good analytical performance with a detection limit of 0.14±0.01 µM and a stable response for more than one month.

A loop suture for dynamic tendon transfer in the repair of chronic or acute extensor pollicis longus ruptures.

Ruptures of extensor pollicis longus (EPL) tendon may need a tendon transfer for repair, especially in chronic ruptures. We describe a novel transfer technique using a loop-tendon suture for dynamic transfer of extensor pollicis brevis to EPL. We performed an end-to-side transfixing-loop self-locking suture preserving the extensor pollicis brevis, which acts as a motor unit for the distal end of the repaired tendon. Contraindications would mainly be simultaneous rupture of EPL and brevis tendons. No complications were observed, except for mild metacarpal-phalangeal stiffness in chronic ruptures. An early rehabilitation program may be started when no other associated injuries exist.

Relationship between physician and industry in Aragon (Spain).

OBJECTIVE: To describe the relationship between industry and physicians and to analyze the physician characteristics associated with the probability of receiving benefits from industry in Aragon (Spain). METHODS: We carried out an observational, cross-sectional study in which Aragonese physicians (north-east region in Spain) from public and private settings completed an anonymous questionnaire on a web page between June and November 2008. Visits/month with industry, samples, gifts, reimbursements and payments were used as dependant variables in the regression analyses. Year of medical license, specialty, work setting, time spent on direct care, articles read/month and being a resident's tutor were used as independent variables. RESULTS: A total of 659 questionnaires were considered valid for the analysis. Overall, 87% (n=573) of the respondents reported they had received some benefit in the previous year and 90.1% (n=593) reported having held meetings with industry representatives monthly. Non-clinical specialists received fewer gifts (odds ratio [OR]=0.38; 95% confidence interval [95%CI]: 0.18-0.77), reimbursements (OR=0.14; 95%CI: 0.06-0.35) and payments (OR=0.30; 95%CI: 0.13-0.74) than their clinical colleagues. The probability of receiving reimbursements (OR=0.37; 95%CI: 0.15-0.89) and payments (OR=0.39; 95%CI: 0.20-0.77) was lower in primary care physicians. CONCLUSIONS: This study, performed in a sample of physicians from a southern European region, demonstrates differences in the intensity of the physician-industry relationship depending on physician specialty and work setting. These results provide important information for improving transparency and for future research on the appropriateness and efficiency of prescription in Spain and other countries with similar health systems.

Oriented immobilization of Desulfovibrio gigas hydrogenase onto carbon electrodes by covalent bonds for nonmediated oxidation of H2.

The orientation of hydrogenase bound covalently to a pyrolytic graphite edge electrode modified with a 4-aminophenyl monolayer can be modulated via electrostatic interactions during the immobilization step. At low ionic strength and when the amino groups of the electrode surface are mostly protonated, the hydrogenase is immobilized with the negatively charged region that surrounds its 4Fe4S cluster nearer to the protein surface facing the electrode. This allows direct electron transfer between the immobilized hydrogenase and the electrode, which is observed by the strong catalytic currents measured in the presence of the H2 substrate. Therefore, a very stable enzymatic electrode is produced that catalyzes nonmediated H2 oxidation.

Functionalization of thioctic acid-capped gold nanoparticles for specific immobilization of histidine-tagged proteins.

This paper presents an efficient strategy for the specific immobilization of fully functional proteins onto the surface of nanoparticles. Thioctic acid-derivatized gold clusters are used as a scaffold for further stepwise modification, leading to a cobalt(II)-terminated ligand shell. A histidine tag introduced by genetic engineering into a protein is coordinated to this transition metal ion. The specific immobilization has been demonstrated for the cases of a genetically engineered horseradish peroxidase and ferredoxin-NADP(+) reductase, confirming the attachment of the fully functional proteins to the Co(II)-terminated nanointerface. The absence of nonspecific protein adsorption and the specificity of the binding site have been verified using several analogues of the enzymes without the histidine tag.