Diagnostic value of urine organic acids analysis among patients with primary mitochondrial disorders

2021 ◽  
pp. 13-24
Author(s):  
Т.Д. Крылова ◽  
М.В. Куркина ◽  
П.В. Баранова ◽  
Е.Ю. Пыркова ◽  
П.Г. Цыганкова ◽  
...  
Keyword(s):  
Organic Acids ◽  
Molecular Genetic ◽  
High Specificity ◽  
Diagnostic Value ◽  
Mitochondrial Disorders ◽  
Test Methods ◽  
Molecular Genetic Analysis ◽  
Defective Structure ◽  
Specificity And Sensitivity ◽  
Urine Organic Acids

Первичные митохондриальные заболевания (ПМЗ) - генетически и клинически гетерогенные заболевания, характеризующиеся нарушением структуры или функций системы окислительного фосфорилирования (OXPHOS), включая электрон-транспортную цепь. Несмотря на успешное применение методов секвенирования нового поколения в диагностике наследственных заболеваний в последнее десятилетие, существует ряд объективных трудностей в интерпретации результатов, особенно при обнаружении новых генов или новых вариантов нуклеотидной последовательности. Анализ биомаркеров, которые являются индикаторами нарушения функций митохондрий, является важным этапом в диагностике многих ПМЗ. Целью данной работы было проведение анализа спектра и концентраций 72 органических кислот в моче методом газовой хроматографии с масс-спектрометнией (ГХ-МС,7890А/5975С, Agilent Technologies, США) в выборке из 84 пациентов с подтвержденным молекулярно-генетическими методами диагнозом ПМЗ и оценка их диагностической значимости. Среди 84 пациентов с ПМЗ, отклонения в спектре органических кислот были выявлены в 78% (66/84) случаев. Уникальный спектр органических кислот наблюдался при митохондриальных гепатопатиях, связанных с мутациями в гене DGUOK: наравне с повышением уровня лактата, пирувата, 3-гидроксибутирата было выявлено повышение концентрации 4-гидроксифениллактата, 4-гидроксифенилпирувата. При анализе ROC-кривых было показано, что диагностическая значимость маркеров убывает в ряду: 3-гидроксибутират, лактат, пируват. При проведении оценки достоверности теста показано, что повышение концентраций пирувата и 4-гидроксифениллактата может быть принято во внимание при предположении ПМЗ у пациента. Introduction. Primary mitochondrial disorders (PMD) are a group of clinically and genetically heterogeneous group of diseases characterized by a defective structure and functions of the Oxidative Phosphorylation System (OXPHOS). Despite the advantages of the next generation sequencing, diagnosis of PMD is still challenging. There is no currently available biomarker with high specificity and sensitivity. But the level of metabolites reflecting the defective OXPHOS is needed for making of a diagnosis of PMD. Aim: to reveal the level and spectrum of urine organic acids among patients with confirmed diagnosis (by molecular-genetic analysis) of PMD and to estimate the diagnostic value of the test. Methods. We measured 72 different metabolites in 84 urine samples from patients with PMD by GC-MS (7890А/5975С, Agilent Technologies, USA). Results. In 66/84 cases among the patients, we detected the abnormal level of urine organic acids. We observed a unique spectrum of metabolites in the patients with DGUOK-associated hepatopathy (abnormal levels of lactate, pyruvate, 3-hydroxybutyrate, and at the same time 4-hydroxyphenyllactate and 4-hydroxyphenylpyruvate). Using ROC-analysis one of the most informative biomarkers was 3-hydroxybutyrate. But due to the lack of specificity, it could not be classified as a valuable biomarker for PMD. The high level of pyruvate and 4-hydroxyphenyllactate could be taken into account to make a diagnosis of PMD

scholarly journals Novel clinical biomarkers in blood and pleural effusion for diagnosing patients with tuberculosis distinguishing from malignant tumor

2021 ◽  
Author(s):  
Jian Wang ◽  
Zhe-Xiang Feng ◽  
Tao Ren ◽  
Wei-Yu Meng ◽  
Imran Khan ◽  
...  
Keyword(s):  
Pleural Effusion ◽  
Malignant Tumor ◽  
Malignant Tumors ◽  
High Specificity ◽  
Diagnostic Value ◽  
Blood Parameters ◽  
Clinical Indicators ◽  
Clinical Biomarkers ◽  
Specificity And Sensitivity ◽  
Single Marker

Abstract Background Pleural effusion (PE) is a common manifestation of tuberculosis and malignant tumors, but it is difficult to distinguish tuberculous pleural effusion (TPE) and malignant pleural effusion (MPE), especially by non-invasive detection indicators. We aimed to find effective detection indexes in blood and PE for differentiating patients with tuberculosis from a malignant tumor. Methods 815 patients were collected who diagnosed with tuberculosis or cancer at Taihe hospital from 2014 to 2017. 717 patients were found to have PE by thoracoscopy. The clinical characteristics, patients’ blood parameters, and PE indicators information were summarized for analysis. Results The patients with MPE had higher percentages to be bloody and negative of Rivalta test in PE than patients with TPE. Then for clinical indicators, comparing specific parameters in blood, we observed 18 indicators were higher in the TPE group than in the MPE group. On contrast, 12 indicators were higher in the MPE group than in the TPE group (p < 0.01). In addition, in PE tests, we found there were 3 parameters higher in TPE and other 4 parameters higher in MPE patients group (p < 0.01). Then for clinical diagnosing practice, ROC and PCA analysis were applied. Top six relevant indicators with AUC value over 0.70 were screened out: pADA (0.90), pHsCRP (0.79), sMONp (0.75), sHsCRP (0.73), sESR (0.71), and sD-dimer (0.70). Moreover, with the Logistic regression model, a specific combination of 3 biomarkers pADA, sMONp, and sHsCRP could enhance distinguishing tuberculosis from malignant tumor patients with PE (AUC=0.944, 95% CI=0.925-0.964). For the top single marker pADA, we further analyzed its diagnostic function in patients with different group and observed it kept the high specificity and sensitivity. Conclusions The six indicators of pADA, pHsCRP, sMONp, sHsCRP, sESR and sD-dimer showed significant diagnostic value for clinicians. Further, the combination of pADA, sMONp, and sHsCRP has high accuracy for differential diagnosis for the first time. Mostly interestingly, pADA single marker maintained high specificity and sensitivity in patients with different status, which has great value for the rapid and accurate diagnosis of suspected cases.

scholarly journals MicroRNA-183-5p acts as a potential diagnostic biomarker for atherosclerosis and regulates the growth of vascular smooth muscle cell

2019 ◽  
Author(s):  
Bin Sun ◽  
Zhengkun Shan ◽  
Guoyu Sun ◽  
Xiaolong Wang
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Roc Analysis ◽  
High Specificity ◽  
Diagnostic Value ◽  
Clinical Value ◽  
Specificity And Sensitivity ◽  
And Migration ◽  
Vsmcs Proliferation ◽  
Proliferation And Migration

Abstract Background Atherosclerosis (AS) is a multifactorial chronic disease, and vascular smooth muscle cells (VSMCs) plays an important role in the pathology of AS. MicroRNAs regulate multiple cellular biological processes. This study aimed to investigate the clinical value of miR-183-5p in AS patients, and further explored the effects of miR-183-5p on the proliferation and migration of VSMCs. Methods qRT-PCR was used to test the level of miR-183-5p. The diagnostic value of miR-183-5p for AS patients was assessed by a receiver operating characteristic (ROC) analysis. Cell proliferation and migration were determined via CCK-8 and Transwell assay. Results MiR-183-5p was highly expressed in AS patients compared with the healthy group. Serum miR-183-5p expression was positively associated with CIMT and CRP in AS patients. The ROC analysis suggested that miR-183-5p had quality to be used as a biomarker with high specificity and sensitivity for AS detection. Overexpression of miR-183-5p promoted the proliferation and migration of VSMCs. Downregulation of miR-183-5p attenuated ox-LDL stimulated VSMCs proliferation and migration. Conclusion MiR-183-5p is highly expressed in AS patients, and downregulation of miR-183-5p attenuated ox-LDL stimulated VSMCs proliferation and migration. MiR-183-5p may be a key molecular for the diagnosis and treatment of AS in the future.

scholarly journals Diagnostic and Prognostic Value of Three microRNAs in Environmental Asbestiform Fibers-Associated Malignant Mesothelioma

2021 ◽  
Vol 11 (11) ◽  
pp. 1205
Author(s):  
Veronica Filetti ◽  
Carla Loreto ◽  
Luca Falzone ◽  
Claudia Lombardo ◽  
Emanuele Cannizzaro ◽  
...  
Keyword(s):  
Malignant Mesothelioma ◽  
Asbestos Exposure ◽  
High Specificity ◽  
Digital Pcr ◽  
Diagnostic Value ◽  
Diagnostic Significance ◽  
Diagnostic Potential ◽  
Specificity And Sensitivity ◽  
Diagnostic And Prognostic Value

Fluoro-edenite (FE) is an asbestiform fiber identified in Biancavilla (Sicily, Italy). Environmental exposure to FE has been associated with a higher incidence of malignant mesothelioma (MM). The present study aimed to validate the predicted diagnostic significance of hsa-miR-323a-3p, hsa-miR-101-3p, and hsa-miR-20b-5p on a subset of MM patients exposed to FE and matched with healthy controls. For this purpose, MM tissues vs. nonmalignant pleura tissues were analyzed through droplet digital PCR (ddPCR) to evaluate differences in the expression levels of the selected miRNAs and their MM diagnostic potential. In addition, further computational analysis has been performed to establish the correlation of these miRNAs with the available online asbestos exposure data and clinic-pathological parameters to verify the potential role of these miRNAs as prognostic tools. ddPCR results showed that the three analyzed miRNAs were significantly down-regulated in MM cases vs. controls. Receiver operating characteristic (ROC) analysis revealed high specificity and sensitivity rates for both hsa-miR-323a-3p and hsa-miR-20b-5p, which thus acquire a diagnostic value for MM. In silico results showed a potential prognostic role of hsa-miR-101-3p due to a significant association of its higher expression and increased overall survival (OS) of MM patients.

scholarly journals MicroRNA-183-5p acts as a potential diagnostic biomarker for atherosclerosis and regulates the growth of vascular smooth muscle cell

2020 ◽  
Author(s):  
Bin Sun ◽  
Zhengkun Shan ◽  
Guoyu Sun ◽  
Xiaolong Wang
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Roc Analysis ◽  
High Specificity ◽  
Diagnostic Value ◽  
Clinical Value ◽  
Specificity And Sensitivity ◽  
And Migration ◽  
Vsmcs Proliferation ◽  
Proliferation And Migration

Abstract Background Atherosclerosis (AS) is a multifactorial chronic disease, and vascular smooth muscle cells (VSMCs) plays an important role in the pathology of AS. MicroRNAs regulate multiple cellular biological processes. This study aimed to investigate the clinical value of miR-183-5p in AS patients, and further explored the effects of miR-183-5p on the proliferation and migration of VSMCs. Methods qRT-PCR was used to test the level of miR-183-5p. The diagnostic value of miR-183-5p for AS patients was assessed by a receiver operating characteristic (ROC) analysis. Cell proliferation and migration were determined via CCK-8 and Transwell assay. Results MiR-183-5p was highly expressed in AS patients compared with the healthy group. Serum miR-183-5p expression was positively associated with CIMT and CRP in AS patients. The ROC analysis suggested that miR-183-5p had quality to be used as a biomarker with high specificity and sensitivity for AS detection. Overexpression of miR-183-5p promoted the proliferation and migration of VSMCs. Downregulation of miR-183-5p attenuated ox-LDL stimulated VSMCs proliferation and migration. Conclusion MiR-183-5p is highly expressed in AS patients, and downregulation of miR-183-5p attenuated ox-LDL stimulated VSMCs proliferation and migration. MiR-183-5p may be a key molecular for the diagnosis and treatment of AS in the future.

scholarly journals The Potential of Steroid Profiling by Mass Spectrometry in the Management of Adrenocortical Carcinoma

Biomedicines ◽  
2020 ◽  
Vol 8 (9) ◽  
pp. 314 ◽  
Author(s):  
Claudia Rossi ◽  
Ilaria Cicalini ◽  
Sara Verrocchio ◽  
Giulia Di Dalmazi ◽  
Luca Federici ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Adrenocortical Carcinoma ◽  
Molecular Genetic ◽  
High Specificity ◽  
Adrenocortical Adenomas ◽  
Specificity And Sensitivity ◽  
Steroid Profiling ◽  
Clinical Dilemma ◽  
Work Up

Radiological and endocrinological work up of adrenal neoplasms is aimed at distinguishing between frequent non-functioning adenomas and rare but very aggressive adrenocortical carcinoma (ACC). Relevant research has addressed the identification of molecular, genetic and hormonal markers that could have clinical significance for malignancy, as well as a prognostic value. Regarding endocrine aspects, attention has been paid to the pattern of steroid secretion that can be affected by altered steroidogenic pathway in ACC. The advent of mass spectrometry techniques has overcome many limitations usually associated with immunoassays, allowing the determination of both common and rarely measured steroids in a single analysis with high specificity and sensitivity. Indeed, mass spectrometry strategies may be able to identify an individualized steroid profile of ACC, allowing a rapid diagnosis and a specific follow-up. In this review, insights, strengths and limitations of mass spectrometry-based approaches in steroid profiling, as well as of immunoassay in steroid measurements, will be specifically discussed. Moreover, the latest findings on steroid profiling by mass spectrometry-based techniques, the most promising analytical tool, will be summarized to evaluate if steroid profiling might be the clue for solving the clinical dilemma in differentiating ACC from non-functioning adrenocortical adenomas (ACA).

scholarly journals Preliminary Evaluation of the Diagnostic Usefulness of Uroplakin 2 with an Assessment of the Antioxidant Potential of Patients with Bladder Cancer

2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
M. Matuszewski ◽  
B. Szymańska ◽  
A. Długosz ◽  
B. Małkiewicz ◽  
J. Dembowski ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
High Specificity ◽  
Diagnostic Value ◽  
Antioxidant Potential ◽  
Antioxidant Systems ◽  
Preliminary Evaluation ◽  
Specificity And Sensitivity ◽  
Total Antioxidant ◽  
Positive Correlations ◽  
And Function

Background. Urothelial carcinoma is the most common type of bladder cancer (BC). It makes up more than 90% of all bladder cancers. Uroplakins are tissue-specific, glycoproteins, playing a role in the construction and function of urothelium. The emergence of uroplakins in the urine and/or plasma may be of potential importance in the early detection of BC. In our study, the diagnostic value of plasma and urine uroplakin 2 (UP2) concentration in bladder cancer was investigated, with an assessment of the antioxidant potential of BC patients. The correlation between UP2, total antioxidant capacity (TAC), and concentration of glutathione (GSH) was also examined. Materials and Methods. This study included 61 BC patients and 33 healthy controls. UP2 concentration was estimated by the immunoenzymatic method (ELISA). TAC and GSH were determined in spectrophotometrically methods. Results. UP2 concentration in BC patients was significantly higher (p≤0.001) both in plasma and in urine compared to the control groups (C). TAC concentration in urine (p≤0.001) and GSH concentration in plasma (p=0.047) were significantly lower in BC group compared to the C group. The high specificity and sensitivity for UPK2 in plasma (76%, 80%, respectively) and urine (88%, 84%, respectively) were observed. Positive correlations were observed between concentration of UP2 in plasma and TAC concentration in urine and between UP2 concentration in plasma and GSH concentration in the same material. Conclusion. The study showed the early diagnostic value of urine and plasma UP2 in BC. There was a decrease in UP2 concentration in the urine of patients with the development of BC. The decrease of antioxidant systems (TAC, GSH) indicates their relationship with the BC process. Based on the obtained results, it is justified to continue the study in a larger group of patients with BC.

scholarly journals Molecular Genetic Analysis in Thyroid Cancer Risk Stratification

2020 ◽  
Vol 16 (26) ◽  
pp. 32-36
Author(s):  
M.O. Rogova ◽  
◽  
N.S. Martirosian ◽  
L.V. Trukhina ◽  
N.B. Paramonova ◽  
...  
Keyword(s):  
Differential Diagnosis ◽  
Thyroid Cancer ◽  
Mirna Expression ◽  
Expression Profiles ◽  
Molecular Genetic ◽  
Diagnostic Value ◽  
Molecular Genetic Analysis ◽  
Cancer Prognosis ◽  
Brafv600e Mutation

Summary: to conduct a comparative analysis of histological samples in the group of benign and malignant thyroid formations using molecular genetic testing: determination of the BRAFV600E mutation, expression of the HMGA2 gene, microRNA (miR, miRNA). To identify possible correlation of these markers with an unfavorable cancer prognosis. Material and methods. The study included 112 patients who underwent surgery for nodular pathology of the thyroid gland. In all patients, the level of 12 miRNA expression was determined by reverse transcription reaction followed by real-time polymerase chain reaction, as well as the presence of BRAFV600E mutation and HMGA2 gene expression. Results. The BRAFV600E mutation can be considered a specific diagnostic marker for papillary thyroid cancer. According to the results of our study, the BRAFV600E mutation was not found in patients with follicular and medullary thyroid cancer, as well as in individuals with benign thyroid formations. The BRAF mutation is not a reliable prognostic marker, since the relationship with aggressive tumor characteristics has not been proved. The model obtained in our study based on the determination of miR7m2 and 451am2 expression levels showed moderate prognostic value. The diagnostic value of our model based on determining the expression level of miR146bm3, 221m2, 375m4, 7m2 was high. Therefore, determining the level of expression of oncogenic miR31, -146b, -155, -357, -551b can be recommended as a differential diagnosis of benign and malignant thyroid formations. Conclusion. In clinical practice, miRNA expression profiles can be used for differential diagnosis of benign and malignant thyroid formations

scholarly journals COMPARISON OF DIFFERENT METHODS OF MOLECULAR-GENETIC ANALYSIS OF SOMATIC MUTATIONS IN K-RAS GENE IN PATIENTS WITH COLORECTAL CANCER

2012 ◽  
Vol 67 (2) ◽  
pp. 35-41 ◽  
Author(s):  
F. A. Amosenko ◽  
I. V. Karpov ◽  
A. V. Polyakov ◽  
S. P. Kovalenko ◽  
V. A. Shamanin ◽  
...  
Keyword(s):  
Molecular Genetic ◽  
False Negative ◽  
Point Mutations ◽  
Real Life ◽  
Molecular Genetic Analysis ◽  
Ras Gene ◽  
Specific Pcr ◽  
Specificity And Sensitivity ◽  
Allele Specific ◽  
Allele Specific Pcr

Two approaches to somatic point mutations in 12 and 13 codones of K-ras gene were analyzed: PCR/SSCP/AСRS/sequencing and allele-specific PCR in the real-life regimen (Russian set «KRAS-7M»). The comparison was carried out on 62 examples of genomic DNA extracted from frozen colon carcinomas, which underwent manual dissection. The results obtained in two attempts were consistent in 95,2% (N=59). Specificity and sensitivity of K-ras mutations detection using «KRAS-7M» set were 100 and 96,4% respectively, and 94,1 and 100% respectievly using PCR/SSCP/AСRS/ automatic sequencing. False positive results were absent when detecting with «KRAS-7M» and accounted for 2 cases (5,9%) when using PCR/SSCP/ AСRS/automatic sequencing. The only false negative response (3,6%) was obtained analyzing mutations using «KRAS-7M».

Phenotyping and Genotyping of Coagulation Factor V Leiden

1996 ◽  
Vol 75 (02) ◽  
pp. 267-269 ◽  
Author(s):  
H Engel ◽  
L Zwang ◽  
H H D M van Vliet ◽  
J J Michiles ◽  
J Stibbe ◽  
...  
Keyword(s):  
Factor V Leiden ◽  
Coagulation Factor ◽  
Diagnostic Value ◽  
Resistance Test ◽  
Amplification Refractory Mutation System ◽  
Factor V ◽  
Chain Reactions ◽  
Apc Resistance ◽  
Specificity And Sensitivity ◽  
Coagulation Factor V

SummaryThe currently used activated Protein C resistance test demonstrated to be of limited diagnostic value for the detection of the mutant Factor V Leiden. Moreover, this assay is not useful for patients under anticoagulant therapy. A modification of the APC resistance test, applying Factor V deficient plasma is described which demonstrates a specificity and sensitivity of 1.0. The superiority of the modified APC resistance test over the existing APC resistance test was verified by genotyping.For that purpose, the Amplification Refractory Mutation System (ARMS) was applied to the detection of the G to A mutation at position 1691 in the gene encoding coagulation Factor V. The mutation at that position could be easily detected by using each of two allele-specific oligonucleotide primers concomitantly with one common primer in two separate polymerase chain reactions, thereby amplifying a fragment of 186 base-pairs of the Factor V gene.

Molecular Characterization and Genetic Diversity Study in F3 Population of Rice

2013 ◽  
Vol 20 (1-2) ◽  
pp. 1-8
Author(s):  
MM Rahman ◽  
L Rahman ◽  
SN Begum ◽  
F Nur
Keyword(s):  
Genetic Distance ◽  
Gene Diversity ◽  
Random Amplified Polymorphic Dna ◽  
Molecular Genetic ◽  
Molecular Genetic Analysis ◽  
Polymorphic Loci ◽  
Rice Genotypes ◽  
High Level ◽  
Genetic Diversity Study ◽  
Diversity Study

Random Amplified Polymorphic DNA (RAPD) assay was initiated for molecular genetic analysis among 13 F3 rice lines and their parents. Four out of 15 decamer random primers were used to amplify genomic DNA and the primers yielded a total of 41 RAPD markers of which 37 were considered as polymorphic with a mean of 9.25 bands per primer. The percentage of polymorphic loci was 90.24. The highest percentage of polymorphic loci (14.63) and gene diversity (0.0714) was observed in 05-6 F3 line and the lowest polymorphic loci (0.00) and gene diversity (0.00) was found in 05-12 and 05-15 F3 lines. So, relatively high level of genetic variation was found in 05-6 F3 line and it was genetically more diverse compared to others. The average co-efficient of gene differentiation (GST) and gene flow (Nm) values across all the loci were 0.8689 and 0.0755, respectively. The UPGMA dendrogram based on the Nei’s genetic distance differentiated the rice genotypes into two main clusters: PNR-519, 05-19, 05-14, 05-12 and 05-17 grouped in cluster 1. On the other hand, Baradhan, 05-9, 05-13, 05-11, 05-5, 05-6, 05-1, 05-4, 05-15 and 05-25 were grouped in cluster 2. The highest genetic distance (0.586) was found between 05-4 and 05-17 F3 lines and they remain in different cluster.DOI: http://dx.doi.org/10.3329/pa.v20i1-2.16839 Progress. Agric. 20(1 & 2): 1 – 8, 2009

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