Analytical validation of an automated assay for the measurement of adenosine deaminase (ADA) and its isoenzymes in saliva and a pilot evaluation of their changes in patients with SARS-CoV-2 infection

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Lorena Franco-Martínez ◽  
Fernando Tecles ◽  
Alberto Torres-Cantero ◽  
Enrique Bernal ◽  
Indra San Lázaro ◽  
...  
Keyword(s):  
Adenosine Deaminase ◽  
Analytical Validation ◽  
Mean Values ◽  
Chemical Inactivation ◽  
Non Invasive ◽  
Automated Assay ◽  
Coefficients Of Variation ◽  
Recovery Percentage ◽  
Triton X ◽  
Potential Use

Abstract Objectives The aim of the present study was to validate a commercially available automated assay for the measurement of total adenosine deaminase (tADA) and its isoenzymes (ADA1 and ADA2) in saliva in a fast and accurate way, and evaluate the possible changes of these analytes in individuals with SARS-CoV-2 infection. Methods The validation, in addition to the evaluation of precision and accuracy, included the analysis of the effects of the main procedures that are currently being used for SARS-CoV-2 inactivation in saliva and a pilot study to evaluate the possible changes in salivary tADA and isoenzymes in individuals infected with SARS-CoV-2. Results The automated assay proved to be accurate and precise, with intra- and inter-assay coefficients of variation below 8.2%, linearity under dilution linear regression with R2 close to 1, and recovery percentage between 80 and 120% in all cases. This assay was affected when the sample is treated with heat or SDS for virus inactivation but tolerated Triton X-100 and NP-40. Individuals with SARS-CoV-2 infection (n=71) and who recovered from infection (n=11) had higher mean values of activity of tADA and its isoenzymes than healthy individuals (n=35). Conclusions tADA and its isoenzymes ADA1 and ADA2 can be measured accurately and precisely in saliva samples in a rapid, economical, and reproducible way and can be analyzed after chemical inactivation with Triton X-100 and NP-40. Besides, the changes observed in tADA and isoenzymes in individuals with COVID-19 open the possibility of their potential use as non-invasive biomarkers in this disease.

scholarly journals Adenosine deaminase activity in pig saliva: analytical validation of two spectrophotometric assays

2017 ◽  
Vol 30 (1) ◽  
pp. 175-179 ◽  
Author(s):  
Fernando Tecles ◽  
Camila P. Rubio ◽  
María D. Contreras-Aguilar ◽  
Marina López-Arjona ◽  
Silvia Martínez-Miró ◽  
...  
Keyword(s):  
Linear Regression ◽  
Confidence Interval ◽  
Adenosine Deaminase ◽  
Analytical Validation ◽  
Altman Plot ◽  
Bland Altman Plot ◽  
Manual Method ◽  
Automated Assay ◽  
Coefficients Of Variation ◽  
Adenosine Deaminase Activity

We validated 2 assays for the measurement of adenosine deaminase (ADA) activity in the saliva of pigs: the Giusti–Galanti manual method (ADA-GG) and a commercial automated assay (Diazyme Laboratories; ADA-D). Intra-assay coefficients of variation (CVs) were <7 and 9%, and interassay CVs were <12 and 5%, for ADA-GG and ADA-D, respectively. Accuracy was measured by 2 methods: recovery and linearity-under-dilution. Recovery was 82.4–106.8% for ADA-GG, and 92.8–107.9% for ADA-D. Serial dilutions showed R2 > 0.95 and 0.99 for ADA-GG and ADA-D, respectively. Linear regression between the methods gave R2 = 0.997 ( p < 0.0001), and a Bland–Altman plot showed a proportional bias of 112 IU/L (95% confidence interval of −99 to 322 IU/L) for ADA-D. No significant differences were observed between the results obtained by either method in saliva or serum. ADA activity was much higher in porcine saliva than in serum. Salivary ADA activity was significantly higher in lame pigs compared to healthy animals. However, serum ADA activity was significantly lower in lame pigs.

scholarly journals Analytical validation of an automated assay for ferric-reducing ability of plasma in dog serum

2017 ◽  
Vol 29 (4) ◽  
pp. 574-578 ◽  
Author(s):  
Camila Peres Rubio ◽  
Silvia Martinez-Subiela ◽  
Josefa Hernández-Ruiz ◽  
Asta Tvarijonaviciute ◽  
José Joaquín Ceron
Keyword(s):  
Detection Limit ◽  
Analytical Validation ◽  
Frap Assay ◽  
Serial Sample ◽  
Automated Assay ◽  
Reducing Ability ◽  
Coefficients Of Variation ◽  
Healthy Dogs ◽  
Interassay Precision

We performed analytical validation of an automated ferric-reducing ability of plasma (FRAP) assay in the serum of dogs. Intra- and interassay precision, accuracy, detection limit, and effects of hemolysis and lipemia were evaluated. Intra- and interassay coefficients of variation were <1% and <13%, respectively. The assay showed a high correlation with a FRAP assay described previously, and results were linear when serial sample dilutions were tested. The detection limit was lower than the values observed in sera from healthy dogs; decreased serum FRAP was found in dogs with leishmaniosis. Lipemia and hemolysis caused a significant increase in the results of the assay.

Role of Cardiac MRI in Detecting Familial Hypertrophic Cardiomyopathy: Review

2016 ◽  
Vol 1 (1) ◽  
pp. 4
Author(s):  
Marymol Koshy ◽  
Bushra Johari ◽  
Mohd Farhan Hamdan ◽  
Mohammad Hanafiah
Keyword(s):  
Hypertrophic Cardiomyopathy ◽  
Magnetic Resonance ◽  
Genetic Disorder ◽  
Familial Hypertrophic Cardiomyopathy ◽  
Non Invasive ◽  
Wide Range ◽  
Proper Diagnosis ◽  
Magnetic Resonance Imaging Mri ◽  
Potential Use

Hypertrophic cardiomyopathy (HCM) is a global disease affecting people of various ethnic origins and both genders. HCM is a genetic disorder with a wide range of symptoms, including the catastrophic presentation of sudden cardiac death. Proper diagnosis and treatment of this disorder can relieve symptoms and prolong life. Non-invasive imaging is essential in diagnosing HCM. We present a review to deliberate the potential use of cardiac magnetic resonance (CMR) imaging in HCM assessment and also identify the risk factors entailed with risk stratification of HCM based on Magnetic Resonance Imaging (MRI).

Potential use of transrenal DNA for non-invasive monitoring and prognosis of colorectal cancer

Biomarkers ◽  
2019 ◽  
Vol 24 (6) ◽  
pp. 524-529
Author(s):  
Wei Chen ◽  
Yingying Liao ◽  
Chunxia Yang ◽  
Zhicheng Fang ◽  
Boyi Liu ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Invasive Monitoring ◽  
Non Invasive ◽  
Potential Use

Potential clinical applications of circulating cell-free DNA in ovarian cancer patients

2018 ◽  
Vol 20 ◽  
Author(s):  
Ana Barbosa ◽  
Ana Peixoto ◽  
Pedro Pinto ◽  
Manuela Pinheiro ◽  
Manuel R. Teixeira
Keyword(s):  
Ovarian Cancer ◽  
Cancer Patients ◽  
Liquid Biopsy ◽  
Clinical Applications ◽  
Epigenetic Alterations ◽  
Cell Free Dna ◽  
Non Invasive ◽  
Free Dna ◽  
Potential Use ◽  
Circulating Cell Free Dna

AbstractCirculating cell-free DNA (cfDNA) consists of small fragments of DNA that circulate freely in the bloodstream. In cancer patients, a fraction of cfDNA is derived from tumour cells, therefore containing the same genetic and epigenetic alterations, and is termed circulating cell-free tumour DNA. The potential use of cfDNA, the so-called ‘liquid biopsy’, as a non-invasive cancer biomarker has recently received a lot of attention. The present review will focus on studies concerning the potential clinical applications of cfDNA in ovarian cancer patients.

Reducing shear strength uncertainties in clays by multivariate correlations

2010 ◽  
Vol 47 (1) ◽  
pp. 16-33 ◽  
Author(s):  
Jianye Ching ◽  
Kok-Kwang Phoon ◽  
Yi-Chu Chen
Keyword(s):  
Shear Strength ◽  
Site Investigation ◽  
Mean Values ◽  
Reliability Based Design ◽  
Economical Design ◽  
Coefficients Of Variation ◽  
Final Design ◽  
Available Information ◽  
Undrained Shear ◽  
Multivariate Correlations

Quantifications of uncertainties in soil shear strengths, including undrained shear strength of clay, are essential for geotechnical reliability-based design. In particular, how to reduce the uncertainties in undrained shear strengths based on all available information by correlation is a practical research subject, given the considerable cost of a typical site investigation. Although it is simple to reduce the uncertainties by correlation when the information is one dimensional (or univariate), it is quite challenging to reduce the uncertainties by using multivariate information through multiple correlations. This study proposes a systematic way of achieving multivariate correlations on undrained shear strengths. A set of simplified equations are obtained through Bayesian analysis for the purpose of reducing uncertainties: the inputs to the equations are the results of in situ or laboratory tests and the outputs are the updated mean values and coefficients of variation (c.o.v.s) of the undrained shear strengths. Two case studies are used to demonstrate the consistency of the proposed simplified equations. Results show that uncertainties in undrained shear strengths can be effectively reduced by incorporating multivariate information. Given that reliability-based design can justify more economical design with reduced uncertainties, the proposed equations essentially link the value of more and better tests directly to final design savings.

scholarly journals Increased Expression of Plasma miRNA-320a and let-7b-5p in Heroin-Dependent Patients and Its Clinical Significance

2021 ◽  
Vol 12 ◽  
Author(s):  
Haixiong Liu ◽  
Wenjin Xu ◽  
Jiying Feng ◽  
Hong Ma ◽  
Jianbin Zhang ◽  
...  
Keyword(s):  
Roc Curves ◽  
Brain Diseases ◽  
Circulating Mirnas ◽  
Heroin Use ◽  
Non Invasive ◽  
Heroin Use Disorder ◽  
Subjective Reports ◽  
The Brain ◽  
Potential Use ◽  
Plasma Mirnas

Heroin use disorder is a chronic and relapsing disease that induces persistent changes in the brain. The diagnoses of heroin use disorders are mainly based on subjective reports and no valid biomarkers available. Recent researches have revealed that circulating miRNAs are useful non-invasive biomarkers for diagnosing brain diseases such as Alzheimer's disease, multiple sclerosis, schizophrenia, and bipolar disorder. However, studies on circulating miRNAs for the diagnosis of heroin use disorders are rarely reported. In this study, we investigated the differential expression of plasma miRNAs in 57 heroin-dependent patients. Based on literature research and microarray analysis, two candidate miRNAs, miR-320a and let-7b-5p, were selected and analyzed by quantitative real-time RT-PCR. The results showed miR-320a and let-7b were significantly upregulated in plasma of the heroin-dependent patients compared to that in healthy controls. The area under curves (AUCs) of receiver operating characteristic (ROC) curves of miR-320a and let-7b-5p were 0.748 and 0.758, respectively. The sensitivities of miR-320a and let-7b-5p were 71.9 and 70.2%, while the specificities of miR-320a and let-7b-5p were 76.1 and 78.3%, respectively. The combination of these two miRNAs predicted heron dependence with an AUC of 0.782 (95% CI 0.687–0.876), with 73.7% sensitivity and 82.6% specificity. Our findings suggest a potential use for circulating miRNAs as biomarkers for the diagnosis of heroin abuse.

scholarly journals Screening of Amazonian bacillus strains for lipase production using glycerol as substrate

ScientiaTec ◽  
2020 ◽  
Vol 7 (03) ◽  
Author(s):  
Giandra Volpato ◽  
Victória Furtado Migliavacca ◽  
Bruna Coelho de Andrade ◽  
Júlio Xandro Heck ◽  
Marco Antônio Záchia Ayub
Keyword(s):  
Organic Synthesis ◽  
Culture Medium ◽  
Lipolytic Activity ◽  
Gum Arabic ◽  
Lipase Production ◽  
Culture Conditions ◽  
Search And Selection ◽  
Triton X ◽  
Potential Use ◽  
Selection Of

The industrial application of lipolytic enzymes has been studied mainly due to the ability of these enzymes in catalyze reactions of synthesis and their stability in various organic solvents. One possibility is the use of lipase the organic synthesis, taking advantage as the generation of waste and difficult recovery of sub bioproducts. In this work, we carried out a selection of eighty-four isolates of Bacillus amazonian for lipase production, of which 30 strains showed lipolytic activity. The study of the culture conditions was performed through a Plackett-Burman experimental design using the strain that presented the highest lipolytic activity in a culture medium using glycerol as substrate.  The studied conditions were: concentration of soybean oil, olive oil, triton X-100, gum arabic, glycerol, and (NH4)2SO4, pH, temperature and concentration of inoculums. The best result obtained were 27 U/L in 48 h of cultivation by Bacillus circulans BL53. This work shows that the search and selection of microorganism with lipolytic activities can facilitate the discovery of new lipases, with potential use as by-product surplus.

scholarly journals Methodological and physiological variability within the ventilatory response to hypoxia in humans

2000 ◽  
Vol 88 (5) ◽  
pp. 1924-1932 ◽  
Author(s):  
Shu Zhang ◽  
Peter A. Robbins
Keyword(s):  
Coefficient Of Variation ◽  
Ventilatory Response ◽  
Systematic Variation ◽  
Hypoxic Ventilatory Response ◽  
Mean Values ◽  
Careful Choice ◽  
Physiological Variability ◽  
Coefficients Of Variation ◽  
Ventilatory Response To Hypoxia ◽  
Over Time

Measurement of the acute hypoxic ventilatory response (AHVR) requires careful choice of the hypoxic stimulus. If the stimulus is too brief, the response may be incomplete; if the stimulus is too long, hypoxic ventilatory depression may ensue. The purpose of this study was to compare three different techniques for assessing AHVR, using different hypoxic stimuli, and also to examine the between-day variability in AHVR. Ten subjects were studied, each on six different occasions, which were ≥1 wk apart. On each occasion, AHVR was assessed using three different protocols: 1) protocol SW, which uses square waves of hypoxia; 2) protocol IS, which uses incremental steps of hypoxia; and 3) protocol RB, which simulates an isocapnic rebreathing test. Mean values for hypoxic sensitivity were 1.02 ± 0.48, 1.15 ± 0.55, and 0.93 ± 0.60 (SD) l ⋅ min− 1 ⋅ %− 1for protocols SW, IS, and RB, respectively. These differed significantly ( P < 0.01). The coefficients of variation for measurement of AHVR were 20, 23, and 36% for the three protocols, respectively. These were not significantly different. There was a significant physiological variation in AHVR ( F 50,100 = 3.9, P < 0.001), with a coefficient of variation of 26%. We conclude that there was relatively little systematic variation between the three protocols but that AHVR varies physiologically over time.

scholarly journals Electrochemical Methodology for Evaluating Skin Oxidative Stress Status (SOSS)

Diseases ◽  
2019 ◽  
Vol 7 (2) ◽  
pp. 40 ◽  
Author(s):  
Pincemail Joël ◽  
Kaci Mouna-Messaouda ◽  
Cheramy-Bien Jean-Paul ◽  
Defraigne Jean-Olivier ◽  
Meziane Smail
Keyword(s):  
Oxidative Stress ◽  
Orange Juice ◽  
Lipid Peroxides ◽  
Oxidative Stress Biomarkers ◽  
Skin Score ◽  
Non Invasive ◽  
Stress Biomarkers ◽  
Coefficients Of Variation ◽  
Oxidative Stress Status ◽  
Copper Zinc

For the purpose of human disease prevention, several methods have been developed, and are still developing, to assess the oxidative stress status (OSS) of individuals. In the present paper, we describe an approach based on electrochemical detection able to evaluate skin oxidative stress status (SOSS) as a PAOT (Pouvoir AntiOxydant Total)-Skin Score®. Normal reference values for the PAOT-Skin Score® were: 0–62.94 (n = 263). Intra- and inter-assay coefficients of variation were, respectively, 12.47 ± 4.29% and 7.0 ± 2.5%. Our technology showed increased skin antioxidant activity following topical application of reduced coeznyme Q10 cream or vitamin C intake as orange juice or supplements. Moreover, we found significant correlations between some blood oxidative stress biomarkers and the PAOT-Skin Score® (γ-tocopherol/α-tocopherol ratio (r = 0.43, p = 0.020); copper (r = −0.42, p = 0.022); copper/zinc ratio (r = −0.49, p = 0.006), and lipid peroxides (r = −0.43, p = 0.002)). In addition to being non–invasive, the present electrochemical methodology is also not expensive, fast, and easy to use.

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