Epithelial-mesenchymal transition-associated microRNA/mRNA signature is linked to metastasis and prognosis in clear-cell renal cell carcinoma

Clear-cell renal cell carcinomas (ccRCCs) are genetically heterogeneous tumors presenting diverse clinical courses. Epithelial-mesenchymal transition (EMT) is a crucial process involved in initiation of metastatic cascade. The aim of our study was to identify an integrated miRNA/mRNA signature associated with metastasis and prognosis in ccRCC through targeted approach based on analysis of miRNAs/mRNAs associated with EMT. A cohort of 230 ccRCC was included in our study and further divided into discovery, training and validation cohorts. EMT markers were evaluated in ccRCC tumor samples, which were grouped accordingly to EMT status. By use of large-scale miRNA/mRNA expression profiling, we identified miRNA/mRNA with significantly different expression in EMT-positive tumors and selected 41 miRNAs/mRNAs for training phase of the study to evaluate their diagnostic and prognostic potential. Fifteen miRNAs/mRNAs were analyzed in the validation phase, where all evaluated miRNA/mRNA candidates were confirmed to be significantly deregulated in tumor tissue. Some of them significantly differed in metastatic tumors, correlated with clinical stage, with Fuhrman grade and with overall survival. Further, we established an EMT- based stage-independent prognostic scoring system enabling identification of ccRCC patients at high-risk of cancer-related death. Finally, we confirmed involvement of miR-429 in EMT regulation in RCC cells in vitro

The measurement of reactive oxygen species in human neat semen and in suspended spermatozoa: a comparison

BACKGROUND: It is generally accepted that oxidative stress is an important factor in male infertility because it may impair the physiological function of spermatozoa at the molecular level. Nevertheless, although several approaches have been reported, the imbalance between production of reactive oxygen species (ROS) and activity of the antioxidant defense system in semen is difficult to investigate and remains poorly understood. METHODS: This study compares measurement of ROS production in neat semen and in washed spermatozoa obtained from the same ejaculate, and suspended in phosphate buffered saline using exactly the same luminol-mediated chemiluminescence method. Ninety one samples were obtained from males of infertile couples and 34 from volunteers with proven fertility. RESULTS: As expected, ROS levels were markedly lower in neat semen than in washed spermatozoa suspensions where seminal plasma with its potent antioxidant capacity was removed. In the cases of both neat semen and washed spermatozoa, ROS production was lowest in samples from normozoospermic males and highest in samples containing more than half million peroxidase-positive leukocytes per milliliter. For all samples, there was a significant positive correlation between ROS production by neat semen and that by washed spermatozoa suspension. CONCLUSION: Measurement of ROS production in neat semen better reflects actual oxidative status because it detects only the overproduction of ROS which are not effectively scavenged by antioxidant capacity of seminal fluid. The results of our study show a good commutability of both measurements for identification of semen samples with high ROS production. The measurement in neat semen is even less time consuming and therefore easier to implement into laboratory routine

Circulating miR-378 and miR-451 in serum are potential biomarkers for renal cell carcinoma

<p>Abstract</p> <p>Background</p> <p>There is no standard serum biomarker used for diagnosis or early detection of recurrence for renal cell carcinoma (RCC) patients. MicroRNAs (miRNAs) are abundant and highly stable in blood serum, and have been recently described as powerful circulating biomarkers in a wide range of solid cancers. Our aim was to identify miRNA signature that can distinguish the blood serum of RCC patients and matched healthy controls and validate identified miRNAs as potential biomarkers for RCC.</p> <p>Methods</p> <p>In the screening phase of the study, blood serum of 15 RCC patients and 12 matched healthy controls were analyzed by use of the TaqMan Low-Density Arrays enabling parallel identification of expression levels of 667 miRNAs through qRT-PCR-based approach. In the validation phase, identified miRNAs were further evaluated on the independent group of 90 RCC patients and 35 matched healthy controls by use of individual qRT-PCR assays and statistically evaluated.</p> <p>Results</p> <p>We identified 30 miRNAs differentially expressed between serum of RCC patients and healthy controls: 19 miRNAs were up-regulated and 11 miRNAs were down-regulated in RCC patients. MiR-378, miR-451 and miR-150 were further evaluated in the independent group of patients, and two of them were successfully validated: levels of miR-378 were increased (p = 0.0003, AUC = 0.71), miR-451 levels were decreased (p < 0.0001, AUC = 0.77) in serum of RCC patients. Combination of miR-378 and miR-451 enable identification of RCC serum with the sensitivity of 81%, specificity 83% and AUC = 0.86.</p> <p>Conclusions</p> <p>Circulating miRNAs in serum are promising biomarkers in RCC.</p

STAT3 and TP53 mutations associate with poor prognosis in anaplastic large cell lymphoma

Funder: European Union’s Horizon 2020 Marie Skłodowska – Curie Innovative Training Networks (ITN - ETN) under grant agreement No.: 675712European Union’s Horizon 2020 Marie Skłodowska – Curie Innovative Training Networks (ITN - ETN) under grant agreement No.: 675712 Czech Science Foundation (GA CR), junior project no. 19-23424Y MEYS CZ project CEITEC 2020 (LQ1601)Funder: MEYS CZ project CEITEC 2020 (LQ1601) NCMG research infrastructure (LM22018132 funded by MEYS CR)Funder: European Union’s Horizon 2020 Marie Skłodowska – Curie Innovative Training Networks (ITN - ETN) under grant agreement No.: 675712.Funder: MH CZ-RVO 6526970

Which health professionals are most at risk for cardiovascular disease? Or do not be a manager

Objectives: Health care workers constitute a high-risk occupational category owing to the character of their work that includes high-risk environment, shift work and mental as well as physical stress. In occupational medicine, caring for their health condition should be a priority and include measures aimed at preventing cardiovascular diseases. The study aimed at determining the prevalence of cardiovascular disease (CVD) risk factors in employees of a large hospital and assessing their effect on the incidence of cardiovascular events. Materials and Methods: The group comprised 3124 employees with a mean age of 36.1 years (SD = 11.4), out of whom 562 were males (mean age of 37.1 years, range: 18-72; SD = 12.26) and 2562 were females (mean age of 35.9 years, range: 18-68; SD = 11.24). At their initial examination, the employees filled in a questionnaire on basic CVD risk factors (according to valid recommendations). This was supplemented with objective data to determine the risk of CVD using valid charts. From this group, a subset of persons at a high or intermediate risk was selected, comprising 247 individuals with a mean age of 54.1 years (SD = 5.73). After 5-9 years (mean 7.24±1.38 years), they either underwent another examination or their health status was ascertained by phone or in a computer database. The end point was the incidence of cardiovascular events (sudden death, acute myocardial infarction, unstable angina pectoris, percutaneous coronary intervention, cardiac failure, stroke or transient ischemic attack). Results: The end point was noted in a total of 15 males (6.07%) and 6 females (2.42%), being statistically significantly present in managers (males p < 0.00007, females p < 0.00001), male physicians/surgeons (p < 0.025), tertiary-educated males (p < 0.0095), female smokers (p < 0.015), male ex-smokers (p < 0.007), overweight or obese males (p < 0.02) and those with the waist-to-hip ratio above 1.0 (p < 0.005). Conclusions: Cardiovascular events are most likely to occur in obese male physicians/surgeons holding managerial positions and in female managers

The measurement of reactive oxygen species in human neat semen and in suspended spermatozoa: a comparison

Abstract Background It is generally accepted that oxidative stress is an important factor in male infertility because it may impair the physiological function of spermatozoa at the molecular level. Nevertheless, although several approaches have been reported, the imbalance between production of reactive oxygen species (ROS) and activity of the antioxidant defense system in semen is difficult to investigate and remains poorly understood. Methods This study compares measurement of ROS production in neat semen and in washed spermatozoa obtained from the same ejaculate, and suspended in phosphate buffered saline using exactly the same luminol-mediated chemiluminescence method. Ninety one samples were obtained from males of infertile couples and 34 from volunteers with proven fertility. Results As expected, ROS levels were markedly lower in neat semen than in washed spermatozoa suspensions where seminal plasma with its potent antioxidant capacity was removed. In the cases of both neat semen and washed spermatozoa, ROS production was lowest in samples from normozoospermic males and highest in samples containing more than half million peroxidase-positive leukocytes per milliliter. For all samples, there was a significant positive correlation between ROS production by neat semen and that by washed spermatozoa suspension. Conclusion Measurement of ROS production in neat semen better reflects actual oxidative status because it detects only the overproduction of ROS which are not effectively scavenged by antioxidant capacity of seminal fluid. The results of our study show a good commutability of both measurements for identification of semen samples with high ROS production. The measurement in neat semen is even less time consuming and therefore easier to implement into laboratory routine.</p

Which health professionals are most at risk for cardiovascular disease? Or do not be a manager

Objectives: Health care workers constitute a high-risk occupational category owing to the character of their work that includes high-risk environment, shift work and mental as well as physical stress. In occupational medicine, caring for their health condition should be a priority and include measures aimed at preventing cardiovascular diseases. The study aimed at determining the prevalence of cardiovascular disease (CVD) risk factors in employees of a large hospital and assessing their effect on the incidence of cardiovascular events. Materials and Methods: The group comprised 3124 employees with a mean age of 36.1 years (SD = 11.4), out of whom 562 were males (mean age of 37.1 years, range: 18-72; SD = 12.26) and 2562 were females (mean age of 35.9 years, range: 18-68; SD = 11.24). At their initial examination, the employees filled in a questionnaire on basic CVD risk factors (according to valid recommendations). This was supplemented with objective data to determine the risk of CVD using valid charts. From this group, a subset of persons at a high or intermediate risk was selected, comprising 247 individuals with a mean age of 54.1 years (SD = 5.73). After 5-9 years (mean 7.24±1.38 years), they either underwent another examination or their health status was ascertained by phone or in a computer database. The end point was the incidence of cardiovascular events (sudden death, acute myocardial infarction, unstable angina pectoris, percutaneous coronary intervention, cardiac failure, stroke or transient ischemic attack). Results: The end point was noted in a total of 15 males (6.07%) and 6 females (2.42%), being statistically significantly present in managers (males p < 0.00007, females p < 0.00001), male physicians/surgeons (p < 0.025), tertiary-educated males (p < 0.0095), female smokers (p < 0.015), male ex-smokers (p < 0.007), overweight or obese males (p < 0.02) and those with the waist-to-hip ratio above 1.0 (p < 0.005). Conclusions: Cardiovascular events are most likely to occur in obese male physicians/surgeons holding managerial positions and in female managers

Bioinformatic pipelines for whole transcriptome sequencing data exploitation in leukemia patients with complex structural variants

Background Extensive genome rearrangements, known as chromothripsis, have been recently identified in several cancer types. Chromothripsis leads to complex structural variants (cSVs) causing aberrant gene expression and the formation of de novo fusion genes, which can trigger cancer development, or worsen its clinical course. The functional impact of cSVs can be studied at the RNA level using whole transcriptome sequencing (total RNA-Seq). It represents a powerful tool for discovering, profiling, and quantifying changes of gene expression in the overall genomic context. However, bioinformatic analysis of transcriptomic data, especially in cases with cSVs, is a complex and challenging task, and the development of proper bioinformatic tools for transcriptome studies is necessary. Methods We designed a bioinformatic workflow for the analysis of total RNA-Seq data consisting of two separate parts (pipelines): The first pipeline incorporates a statistical solution for differential gene expression analysis in a biologically heterogeneous sample set. We utilized results from transcriptomic arrays which were carried out in parallel to increase the precision of the analysis. The second pipeline is used for the identification of de novo fusion genes. Special attention was given to the filtering of false positives (FPs), which was achieved through consensus fusion calling with several fusion gene callers. We applied the workflow to the data obtained from ten patients with chronic lymphocytic leukemia (CLL) to describe the consequences of their cSVs in detail. The fusion genes identified by our pipeline were correlated with genomic break-points detected by genomic arrays. Results We set up a novel solution for differential gene expression analysis of individual samples and de novo fusion gene detection from total RNA-Seq data. The results of the differential gene expression analysis were concordant with results obtained by transcriptomic arrays, which demonstrates the analytical capabilities of our method. We also showed that the consensus fusion gene detection approach was able to identify true positives (TPs) efficiently. Detected coordinates of fusion gene junctions were in concordance with genomic breakpoints assessed using genomic arrays. Discussion Byapplying our methods to real clinical samples, we proved that our approach for total RNA-Seq data analysis generates results consistent with other genomic analytical techniques. The data obtained by our analyses provided clues for the study of the biological consequences of cSVs with far-reaching implications for clinical outcome and management of cancer patients. The bioinformatic workflow is also widely applicable for addressing other research questions in different contexts, for which transcriptomic data are generated