Circulating microRNAs in sera correlate with soluble biomarkers of immune activation but do not predict mortality in ART treated individuals with HIV-1 infection: A case control study

Introduction: The use of anti-retroviral therapy (ART) has dramatically reduced HIV-1 associated morbidity and mortality. However, HIV-1 infected individuals have increased rates of morbidity and mortality compared to the non-HIV-1 infected population and this appears to be related to end-organ diseases collectively referred to as Serious Non-AIDS Events (SNAEs). Circulating miRNAs are reported as promising biomarkers for a number of human disease conditions including those that constitute SNAEs. Our study sought to investigate the potential of selected miRNAs in predicting mortality in HIV-1 infected ART treated individuals. Materials and Methods: A set of miRNAs was chosen based on published associations with human disease conditions that constitute SNAEs. This case: control study compared 126 cases (individuals who died whilst on therapy), and 247 matched controls (individuals who remained alive). Cases and controls were ART treated participants of two pivotal HIV-1 trials. The relative abundance of each miRNA in serum was measured, by RTqPCR. Associations with mortality (all-cause, cardiovascular and malignancy) were assessed by logistic regression analysis. Correlations between miRNAs and CD4+ T cell count, hs-CRP, IL-6 and D-dimer were also assessed. Results: None of the selected miRNAs was associated with all-cause, cardiovascular or malignancy mortality. The levels of three miRNAs (miRs -21, -122 and -200a) correlated with IL-6 while miR-21 also correlated with D-dimer. Additionally, the abundance of miRs -31, -150 and -223, correlated with baseline CD4+ T cell count while the same three miRNAs plus miR- 145 correlated with nadir CD4+ T cell count. Discussion: No associations with mortality were found with any circulating miRNA studied. These results cast doubt onto the effectiveness of circulating miRNA as early predictors of mortality or the major underlying diseases that contribute to mortality in participants treated for HIV-1 infection

Development and Validation of a Risk Score for Chronic Kidney Disease in HIV Infection Using Prospective Cohort Data from the D:A:D Study

Ristola M. on työryhmien DAD Study Grp ; Royal Free Hosp Clin Cohort ; INSIGHT Study Grp ; SMART Study Grp ; ESPRIT Study Grp jäsen.Background Chronic kidney disease (CKD) is a major health issue for HIV-positive individuals, associated with increased morbidity and mortality. Development and implementation of a risk score model for CKD would allow comparison of the risks and benefits of adding potentially nephrotoxic antiretrovirals to a treatment regimen and would identify those at greatest risk of CKD. The aims of this study were to develop a simple, externally validated, and widely applicable long-term risk score model for CKD in HIV-positive individuals that can guide decision making in clinical practice. Methods and Findings A total of 17,954 HIV-positive individuals from the Data Collection on Adverse Events of Anti-HIV Drugs (D:A:D) study with >= 3 estimated glomerular filtration rate (eGFR) values after 1 January 2004 were included. Baseline was defined as the first eGFR > 60 ml/min/1.73 m2 after 1 January 2004; individuals with exposure to tenofovir, atazanavir, atazanavir/ritonavir, lopinavir/ritonavir, other boosted protease inhibitors before baseline were excluded. CKD was defined as confirmed (>3 mo apart) eGFR In the D:A:D study, 641 individuals developed CKD during 103,185 person-years of follow-up (PYFU; incidence 6.2/1,000 PYFU, 95% CI 5.7-6.7; median follow-up 6.1 y, range 0.3-9.1 y). Older age, intravenous drug use, hepatitis C coinfection, lower baseline eGFR, female gender, lower CD4 count nadir, hypertension, diabetes, and cardiovascular disease (CVD) predicted CKD. The adjusted incidence rate ratios of these nine categorical variables were scaled and summed to create the risk score. The median risk score at baseline was -2 (interquartile range -4 to 2). There was a 1: 393 chance of developing CKD in the next 5 y in the low risk group (risk score = 5, 505 events), respectively. Number needed to harm (NNTH) at 5 y when starting unboosted atazanavir or lopinavir/ritonavir among those with a low risk score was 1,702 (95% CI 1,166-3,367); NNTH was 202 (95% CI 159-278) and 21 (95% CI 19-23), respectively, for those with a medium and high risk score. NNTH was 739 (95% CI 506-1462), 88 (95% CI 69-121), and 9 (95% CI 8-10) for those with a low, medium, and high risk score, respectively, starting tenofovir, atazanavir/ritonavir, or another boosted protease inhibitor. The Royal Free Hospital Clinic Cohort included 2,548 individuals, of whom 94 individuals developed CKD (3.7%) during 18,376 PYFU (median follow-up 7.4 y, range 0.3-12.7 y). Of 2,013 individuals included from the SMART/ESPRIT control arms, 32 individuals developed CKD (1.6%) during 8,452 PYFU (median follow-up 4.1 y, range 0.6-8.1 y). External validation showed that the risk score predicted well in these cohorts. Limitations of this study included limited data on race and no information on proteinuria. Conclusions Both traditional and HIV-related risk factors were predictive of CKD. These factors were used to develop a risk score for CKD in HIV infection, externally validated, that has direct clinical relevance for patients and clinicians to weigh the benefits of certain antiretrovirals against the risk of CKD and to identify those at greatest risk of CKD.Peer reviewe

Generic Assay of Sulfur-Containing Compounds Based on Kinetics Inhibition of Gold Nanoparticle Photochemical Growth

This work describes a new, equipment-free, generic method for the determination of sulfur-containing compounds that is based on their ability to slow down the photoreduction kinetics of gold ions to gold nanoparticles. The method involves tracking the time required for a red coloration to appear in the tested sample, indicative of the formation of gold nanoparticles, and compare the measured time relative to a control sample in the absence of the target analyte. The method is applicable with minimal and simple steps requiring only two solutions (i.e., a buffer and a gold solution), a source of light (UV or visible), and a timer. The method responds to a large variety of sulfur-containing compounds including thiols, thioesters, disulfides, thiophosphates, metal–sulfur bonds, and inorganic sulfur and was therefore applied to the determination of a variety of compounds such as dithiocarbamate and organophosphorous pesticides, biothiols, pharmaceutically active compounds, and sulfides in different samples such as natural waters and wastewater, biological fluids, and prescription drugs. The analytical figures of merit of the method include satisfactory sensitivity (quantitation limits at the low μM levels), good recoveries (from 93 to 109%), and satisfactory reproducibility (from 4.8 to 9.8%). The method is easily adoptable to both laboratory settings and nonlaboratory conditions for quantitative and semiquantitative analysis, respectively, is user-friendly even for the minimally trained user, and can be performed with limited resources at low cost

A Low-Cost Colorimetric Assay for the Analytical Determination of Copper Ions with Consumer Electronic Imaging Devices in Natural Water Samples

This study reports a new approach for the determination of copper ions in water samples that exploits the complexation reaction with diethyldithiocarbamate (DDTC) and uses widely available imaging devices (i.e., flatbed scanners or smartphones) as detectors. Specifically, the proposed approach is based on the ability of DDTC to bind to copper ions and form a stable Cu-DDTC complex with a distinctive yellow color detected with the camera of a smartphone in a 96-well plate. The color intensity of the formed complex is linearly proportional to the concentration of copper ions, resulting in its accurate colorimetric determination. The proposed analytical procedure for the determination of Cu2+ was easy to perform, rapid, and applicable with inexpensive and commercially available materials and reagents. Many parameters related to such an analytical determination were optimized, and a study of interfering ions present in the water samples was also carried out. Additionally, even low copper levels could be noticed by the naked eye. The assay performed was successfully applied to the determination of Cu2+ in river, tap, and bottled water samples with detection limits as low as 1.4 µM, good recoveries (89.0–109.6%), adequate reproducibility (0.6–6.1%), and high selectivity over other ions present in the water samples

Ultratrace Determination of Silver, Gold, and Iron Oxide Nanoparticles by Micelle Mediated Preconcentration/Selective Back-Extraction Coupled with Flow Injection Chemiluminescence Detection

A new method has been developed for the ultrasensitive determination of silver, gold, and iron oxide nanoparticles in environmental samples. Cloud point extraction was optimized and used as a means to extract and preconcentrate all nanoparticle species simultaneously from the same sample. The extracted nanoparticles were sequentially isolated from the surfactant-rich phase by a new selective back-extraction procedure and dissociated into their precursor metal ions. Each ion solution was injected in a flow injection analysis (FIA) manifold, accommodating the chemiluminogenic oxidation of luminol, in order to amplify chemiluminescence (CL) emission in a manner proportional to its concentration. Under the optimum experimental conditions, the detection limits were brought down to the picomolar and femtomolar concentration levels with satisfactory analytical features in terms of precision (2.0–13.0%), selectivity against dissolved ions, and recoveries (74–114%). The method was successfully applied to the determination of iron oxide, silver, and gold nanoparticles in environmental samples of different complexity, ranging from unpolluted river water to raw sewage. The developed method could also serve as a basis for future deployment of molecular spectrometry detectors for the selective determination and speciation analysis of nanoparticles in environmental applications

Programming Fluid Transport in Paper-Based Microfluidic Devices Using Razor-Crafted Open Channels

Manipulating fluid transport in microfluidic, paper-based analytical devices (μPADs) is an essential prerequisite to enable multiple timed analytical steps on the same device. Current methods to control fluid distribution mainly rely on controlling how slowly the fluid moves within a device or by activating an on/off switch to flow. In this Article, we present an easy approach for programming fluid transport within paper-based devices that enables both acceleration as well as delay of fluid transport without active pumping. Both operations are programmed by carving open channels either longitudinally or perpendicularly to the flow path using a craft-cutting tool equipped with a knife blade. Channels are crafted after μPADs fabrication enabling the end user to generate patterns of open-channels on demand by carving the porous material of the paper without cutting or removing the paper substrate altogether. Parameters to control the acceleration or delay of flow include the orientation, length, and number of open channels. Using this method, accelerated as well as reduced fluid transport rates were achieved on the same device. This methodology was applied to μPADs for multiple and time-programmable assays for metal ion determination

Solid ink-printed filter paper as a green adsorbent material for the solid-phase extraction of UV filters from water samples

<p>This work describes the development of a new green solid-phase extraction approach, which is based on the use of low-cost extraction discs composed of plain filter papers that are covered with a synthetic wax-like coating. The filter papers are printed in a commercial solid ink printer, which dispenses a synthetic wax-like ink on the surface of the paper, to cover the hydrophilic cellulose fibre matrix with an interface of lipophilic domains where non-polar analytes can partition through hydrophobic interactions. The modified paper filters were used to extract hydrophobic organic compounds from water samples following the customary procedure of solid-phase extraction without sorbent preconditioning and needless of high-vacuum sources. As a proof-of-concept application, a series of non-polar organic UV filters were used as model analytes to optimise the extraction parameters and evaluate the performance of the method in spiked water samples. Based on this principle, a new sample preparation platform with low environmental footprint has been developed that enables extraction to be carried out using low-cost, environmental benign and non-toxic conventional materials. The advantages and disadvantages of the method, alongside with its future prospects towards the development of custom-made ‘printed extraction kits’, are envisioned and discussed.</p