Low-dose adenosine stress echocardiography: Detection of myocardial viability

OBJECTIVE: The aim of this study was to evaluate the diagnostic potential of low-dose adenosine stress echocardiography in detection of myocardial viability. BACKGROUND: Vasodilation through low dose dipyridamole infusion may recruit contractile reserve by increasing coronary flow or by increasing levels of endogenous adenosine. METHODS: Forty-three patients with resting dyssynergy, due to previous myocardial infarction, underwent low-dose adenosine (80, 100, 110 mcg/kg/min in 3 minutes intervals) echocardiography test. Gold standard for myocardial viability was improvement in systolic thickening of dyssinergic segments of ≥ 1 grade at follow-up. Coronary angiography was done in 41 pts. Twenty-seven patients were revascularized and 16 were medically treated. Echocardiographic follow up data (12 ± 2 months) were available in 24 revascularized patients. RESULTS: Wall motion score index improved from rest 1.55 ± 0.30 to 1.33 ± 0.26 at low-dose adenosine (p < 0.001). Of the 257 segments with baseline dyssynergy, adenosine echocardiography identified 122 segments as positive for viability, and 135 as necrotic since no improvement of systolic thickening was observed. Follow-up wall motion score index was 1.31 ± 0.30 (p < 0.001 vs. rest). The sensitivity of adenosine echo test for identification of viable segments was 87%, while specificity was 95%, and diagnostic accuracy 90%. Positive and negative predictive values were 97% and 80%, respectively. CONCLUSION: Low-dose adenosine stress echocardiography test has high diagnostic potential for detection of myocardial viability in the group of patients with left ventricle dysfunction due to previous myocardial infarction. Low dose adenosine stress echocardiography may be adequate alternative to low-dose dobutamine test for evaluation of myocardial viability

Solve-RD: systematic pan-European data sharing and collaborative analysis to solve rare diseases.

For the first time in Europe hundreds of rare disease (RD) experts team up to actively share and jointly analyse existing patient's data. Solve-RD is a Horizon 2020-supported EU flagship project bringing together >300 clinicians, scientists, and patient representatives of 51 sites from 15 countries. Solve-RD is built upon a core group of four European Reference Networks (ERNs; ERN-ITHACA, ERN-RND, ERN-Euro NMD, ERN-GENTURIS) which annually see more than 270,000 RD patients with respective pathologies. The main ambition is to solve unsolved rare diseases for which a molecular cause is not yet known. This is achieved through an innovative clinical research environment that introduces novel ways to organise expertise and data. Two major approaches are being pursued (i) massive data re-analysis of >19,000 unsolved rare disease patients and (ii) novel combined -omics approaches. The minimum requirement to be eligible for the analysis activities is an inconclusive exome that can be shared with controlled access. The first preliminary data re-analysis has already diagnosed 255 cases form 8393 exomes/genome datasets. This unprecedented degree of collaboration focused on sharing of data and expertise shall identify many new disease genes and enable diagnosis of many so far undiagnosed patients from all over Europe

Solving patients with rare diseases through programmatic reanalysis of genome-phenome data.

Funder: EC | EC Seventh Framework Programm | FP7 Health (FP7-HEALTH - Specific Programme "Cooperation": Health); doi: https://doi.org/10.13039/100011272; Grant(s): 305444, 305444Funder: Ministerio de Economía y Competitividad (Ministry of Economy and Competitiveness); doi: https://doi.org/10.13039/501100003329Funder: Generalitat de Catalunya (Government of Catalonia); doi: https://doi.org/10.13039/501100002809Funder: EC | European Regional Development Fund (Europski Fond za Regionalni Razvoj); doi: https://doi.org/10.13039/501100008530Funder: Instituto Nacional de Bioinformática ELIXIR Implementation Studies Centro de Excelencia Severo OchoaFunder: EC | EC Seventh Framework Programm | FP7 Health (FP7-HEALTH - Specific Programme "Cooperation": Health)Reanalysis of inconclusive exome/genome sequencing data increases the diagnosis yield of patients with rare diseases. However, the cost and efforts required for reanalysis prevent its routine implementation in research and clinical environments. The Solve-RD project aims to reveal the molecular causes underlying undiagnosed rare diseases. One of the goals is to implement innovative approaches to reanalyse the exomes and genomes from thousands of well-studied undiagnosed cases. The raw genomic data is submitted to Solve-RD through the RD-Connect Genome-Phenome Analysis Platform (GPAP) together with standardised phenotypic and pedigree data. We have developed a programmatic workflow to reanalyse genome-phenome data. It uses the RD-Connect GPAP's Application Programming Interface (API) and relies on the big-data technologies upon which the system is built. We have applied the workflow to prioritise rare known pathogenic variants from 4411 undiagnosed cases. The queries returned an average of 1.45 variants per case, which first were evaluated in bulk by a panel of disease experts and afterwards specifically by the submitter of each case. A total of 120 index cases (21.2% of prioritised cases, 2.7% of all exome/genome-negative samples) have already been solved, with others being under investigation. The implementation of solutions as the one described here provide the technical framework to enable periodic case-level data re-evaluation in clinical settings, as recommended by the American College of Medical Genetics

Solving unsolved rare neurological diseases-a Solve-RD viewpoint.

Funder: Durch Princess Beatrix Muscle Fund Durch Speeren voor Spieren Muscle FundFunder: University of Tübingen Medical Faculty PATE programFunder: European Reference Network for Rare Neurological Diseases | 739510Funder: European Joint Program on Rare Diseases (EJP-RD COFUND-EJP) | 44140962

A Solve-RD ClinVar-based reanalysis of 1522 index cases from ERN-ITHACA reveals common pitfalls and misinterpretations in exome sequencing

Purpose Within the Solve-RD project (https://solve-rd.eu/), the European Reference Network for Intellectual disability, TeleHealth, Autism and Congenital Anomalies aimed to investigate whether a reanalysis of exomes from unsolved cases based on ClinVar annotations could establish additional diagnoses. We present the results of the “ClinVar low-hanging fruit” reanalysis, reasons for the failure of previous analyses, and lessons learned. Methods Data from the first 3576 exomes (1522 probands and 2054 relatives) collected from European Reference Network for Intellectual disability, TeleHealth, Autism and Congenital Anomalies was reanalyzed by the Solve-RD consortium by evaluating for the presence of single-nucleotide variant, and small insertions and deletions already reported as (likely) pathogenic in ClinVar. Variants were filtered according to frequency, genotype, and mode of inheritance and reinterpreted. Results We identified causal variants in 59 cases (3.9%), 50 of them also raised by other approaches and 9 leading to new diagnoses, highlighting interpretation challenges: variants in genes not known to be involved in human disease at the time of the first analysis, misleading genotypes, or variants undetected by local pipelines (variants in off-target regions, low quality filters, low allelic balance, or high frequency). Conclusion The “ClinVar low-hanging fruit” analysis represents an effective, fast, and easy approach to recover causal variants from exome sequencing data, herewith contributing to the reduction of the diagnostic deadlock

Twist exome capture allows for lower average sequence coverage in clinical exome sequencing

Background Exome and genome sequencing are the predominant techniques in the diagnosis and research of genetic disorders. Sufficient, uniform and reproducible/consistent sequence coverage is a main determinant for the sensitivity to detect single-nucleotide (SNVs) and copy number variants (CNVs). Here we compared the ability to obtain comprehensive exome coverage for recent exome capture kits and genome sequencing techniques. Results We compared three different widely used enrichment kits (Agilent SureSelect Human All Exon V5, Agilent SureSelect Human All Exon V7 and Twist Bioscience) as well as short-read and long-read WGS. We show that the Twist exome capture significantly improves complete coverage and coverage uniformity across coding regions compared to other exome capture kits. Twist performance is comparable to that of both short- and long-read whole genome sequencing. Additionally, we show that even at a reduced average coverage of 70× there is only minimal loss in sensitivity for SNV and CNV detection. Conclusion We conclude that exome sequencing with Twist represents a significant improvement and could be performed at lower sequence coverage compared to other exome capture techniques

Development and Application of Ligand-Exchange Reaction Method for the Determination of Clonazepam

Purpose: This paper presents an improved kinetic-spectrophotometric procedure for determining clonazepam (CZP) in pharmaceutical formulations and human serum. Methods: The method is based on ligand-exchange reaction. The reaction was followed spectrophotometrically by measuring the rate of change of absorbance at 425 nm in ethanolic sodium hydroxide solution.3 Results: The optimum operating conditions for reagent concentrations and temperature were established. Linear calibration curve was obtained in the range of 0.32 - 4.10 μg mL-1. The optimized conditions yielded a theoretical detection limit of 0.24 μg mL-1 based on the 3.3So criterion, where S0 is standard deviation of the calibration line. The interference of certain drugs, foreign ions and amino acids on the reaction rate were studied in order to assess the selectivity of the method. Conclusion: The developed method is sensitive, accurate and reproducible and could be used for routine anlysis of clonazepam in pharmaceutical preparations and serum samples

Evaluation of individual phenolic compounds and antioxidant properties of black, green, herbal and fruit tea infusions consumed in Serbia: spectrophotometrical and electrochemical approaches

The aim of this study was evaluation of individual phenolic compounds and antioxidant activity of commercially consumed black, green, fruit and herbal tea infusions in Serbia in order to characterize the quantity and quality of teas. The most abundant compound was gallic acid, followed by caffeic acid, rutin, (+)-catechin and (-)-epicatechin. The main procyanidin was procyanidin B1. The antioxidant activity was measured using five in vitro methods: determination of 1,1-dipheny1-2-picrylhydrazyl radical-scavenging activity (DPPH), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) radical cation-scavenging activity (ABTS), ferric reducing antioxidant power (FRAP), reduction power (RP) Fe(III) to Fe(II) and cyclic voltammetry (CV). Obtained results of FRAP and of the Fe(III)/Fe(III) method correlated strongly with the total phenolics content (R-2 = 0.92246, R-2 = 0.88084, p lt 0.0001). Antioxidant power of green tea and bearberry tea was considerably higher than that of black tea. Raspberry and cherry showed the highest antioxidant power among fruit tea infusions. Contribution of phenolic compounds to tea antioxidant activity was also quantified in this study. Stepwise linear regression demonstrated that quantification of different phenolic compounds responsible for tea antioxidant activity was dependent on the method used. Gallic acid, caffeic acid (+)-catechin, (-)-epicatechin, (-)-epigallocatechin, procyanidin B1, procyanidin B2 together made up 43.6-99.9% of the antioxidant activity of tea

Evaluation of individual phenolic compounds and antioxidant properties of black, green, herbal and fruit tea infusions consumed in Serbia: spectrophotometrical and electrochemical approaches

The aim of this study was evaluation of individual phenolic compounds and antioxidant activity of commercially consumed black, green, fruit and herbal tea infusions in Serbia in order to characterize the quantity and quality of teas. The most abundant compound was gallic acid, followed by caffeic acid, rutin, (+)-catechin and (-)-epicatechin. The main procyanidin was procyanidin B1. The antioxidant activity was measured using five in vitro methods: determination of 1,1-dipheny1-2-picrylhydrazyl radical-scavenging activity (DPPH), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) radical cation-scavenging activity (ABTS), ferric reducing antioxidant power (FRAP), reduction power (RP) Fe(III) to Fe(II) and cyclic voltammetry (CV). Obtained results of FRAP and of the Fe(III)/Fe(III) method correlated strongly with the total phenolics content (R-2 = 0.92246, R-2 = 0.88084, p lt 0.0001). Antioxidant power of green tea and bearberry tea was considerably higher than that of black tea. Raspberry and cherry showed the highest antioxidant power among fruit tea infusions. Contribution of phenolic compounds to tea antioxidant activity was also quantified in this study. Stepwise linear regression demonstrated that quantification of different phenolic compounds responsible for tea antioxidant activity was dependent on the method used. Gallic acid, caffeic acid (+)-catechin, (-)-epicatechin, (-)-epigallocatechin, procyanidin B1, procyanidin B2 together made up 43.6-99.9% of the antioxidant activity of tea