Marcin Dziedzic

The diagnostic role of plasma circulating precursors of miRNA-944 and miRNA-3662 for non-small cell lung cancer detection

INTRODUCTION microRNA (miRNA) seem to be most attractive cancer markers due their crucial role in tumor development and possibility of their analysis using liquid biopsy. To date there is little known about role of miRNA precursors (pri-miRNA) in carcinogenesis and their utility as tumor markers. MATERIAL AND METHODS miRNA-944 and miRNA-3662 precursors as potential non-small cell lung cancer (NSCLC) markers were analyzed in plasma samples of 56 patients in an early stage of NSCLC and 100 healthy individuals. RESULTS Diagnostic test based on two studied markers for stage I-IIIA of the disease allowed to distinguish NSCLC from healthy individuals with 75.7% sensitivity and 82.3% specificity (AUC=0.898). pri-miRNA-944 distinguished SCC from AC with sensitivity of 78.6% and specificity of 91.7% (AUC=0.771), and pri-miRNA-3662 distinguished AC from SCC with 57.1% sensitivity and 90% specificity (AUC=0.845). CONCLUSION Circulating pri-miRNA-944 and 3662 can improve non-invasive NSCLC detection of operable stages of SCC and AC. miRNA precursors could be considered as novel potential lung cancer biomarkers.

Cholinergic activation affects the acute and chronic antinociceptive effects of morphine

Current studies indicate that the cholinergic and opioid systems interact to modulate pain. In the present work, we investigated the influence of the cholinesterase inhibitors, donepezil (0.5; 1 or 3mg/kg, i.p.) and rivastigmine (0.03; 0.5 or 1mg/kg, i.p.), on the acute antinociceptive effects of morphine (5mg/kg, i.p.) in the hot plate test in mice. Herein, both inhibitors were found to enhance and prolong the analgesic effects of morphine without affecting latencies themselves. In an extension of this work, we determined which cholinergic receptors subtype mediates the enhancement of analgesic effects of morphine, following inhibition of cholinesterases. In this part of the study, scopolamine (0.5mg/kg, i.p.), a muscarinic cholinergic receptors antagonist, but not mecamylamine (3mg/kg, i.p.), a nicotinic cholinergic receptors antagonist, reversed the enhancing effects of donepezil (3mg/kg, i.p.) and rivastigmine (1mg/kg, i.p.) on the morphine antinociception. Moreover, both cholinesterase inhibitors attenuated the development of tolerance to the antinociceptive effects of morphine. In contrast, acute administration of donepezil (3mg/kg, i.p.) or rivastigmine (1mg/kg, i.p.) on the day of expression of tolerance, had no effect on the already developed morphine tolerance. What is more, in both set of experiments, rivastigmine was slightly more potent than donepezil due to the broader inhibitory spectrum of this drug on acetylcholine degradation. Thus, our results suggest that the cholinesterase inhibitors, donepezil and rivastigmine, may be administered with morphine in order to enhance the latters analgesic effects for the treatment of acute and chronic pain.

Impact of I/D polymorphism of ACE gene on risk of development and course of chronic obstructive pulmonary disease

Introduction Chronic obstructive pulmonary disease (COPD) affects more than 10% of the worlds population over 40 years of age. The main exogenous risk factor is cigarette smoking; however, only 20% of smokers develop COPD, indicating that some other factors, e.g. genetic, may play an important role in the disease pathogenesis. Recent research indicates that ACE (angiotensin-converting enzyme) may be a susceptibility gene for asthma or COPD. The aim of our study was to determine the influence of I/D (insertion/deletion) polymorphism of the ACE gene (AluYa5, rs4646994) on the risk and course of COPD. Material and methods We investigated ACE I/D polymorphism in 206 COPD and 165 healthy Caucasian subjects. Results In the generalized linear model (GLZ) analysis of the influence of selected factors on presence of COPD we found a significant independent effect for male sex (repeatedly increases the risk of COPD, OR = 7.7, p = 0.049), as well as smoking or lower body mass index, but only in combination with older age (OR = 0.96, p = 0.003 and OR = 1.005, p = 0.04 respectively). Interestingly, analysis of factors which may influence the risk of a higher number of exacerbations demonstrated that occurrence of DD genotype, but only in men, is associated with a lower risk (OR = 0.7, p = 0.03) of this complication. Conclusions We suggest that ACE may not be a susceptibility gene for the origin of COPD but a disease-modifying gene. Since the impact of I/D polymorphism of the ACE gene on COPD risk is moderate or negligible, other molecular changes, that will help predict the development of this disease, should still be sought.

Omentin - a new adipokine with many roles to play

Abstract Adipose tissue is at a point of high interest in medical research, not only as an energy depot, but also because it secretes nearly more than 600 cytokines. These are termed‚ adipokines’. Human adipokines are involved in numerous metabolic processes, including the regulation of appetite, energy expenditure, insulin sensitivity, inflammation and cardiovascular activity. Thus, these could be clinically important as a markers of adipose tissue function and increased metabolic risk. The search for novel adipokines linking obesity to related co-morbidities has become a major topic in obesity research. In such work, there is an increasing need to define their function, their molecular targets and their potential clinical relevance as biomarkers or in the treatment of obesity and other metabolic diseases. Omentin (34 kDa) is a recently identified fat deposition-specific adipokine with multiple interactions. Concentrations of omentin have been shown to be decreased in patients with obesity and impaired glucose regulation, in patients afflicted with diabetes type 1 and 2, and in patients with polycystic ovary syndrome. These are all diseases commonly associated with insulin resistance and obesity. The aim of this study was to show and compare the latest information about omentin and its relationships with obesity, diabetes mellitus (DM), metabolic syndrome (MetS), inflammation, cardiac problems, sex hormone imbalances and cancer. The association of omentin with particular metabolic indexes may suggest that an elevation in omentin level may be seen as being a marker for leanness, while a decreased level will underline possible situations of overweight and obesity along with their comorbidities (diabetes, cardiovascular disease, metabolic syndrome, inflammation and even cancer). However, a challenge for the future is to fully understand the multiple role played by omentin. Thus, more studies in these matter are required.

Relationship between microRNA-146a expression and plasma renalase levels in hemodialyzed patients

Background microRNA (miRNA) belongs to the non-coding RNAs family responsible for the regulation of gene expression. Renalase is a protein composed of 342 amino acids, secreted by the kidneys and possibly plays an important role in the regulation of sympathetic tone and blood pressure. The aim of the present study was to investigate plasma renalase concentration, and explore the relationship between miRNA-146a-5p expression and plasma renalase levels in hemodialyzed patients. Methods The study population comprised 55 subjects who succumbed to various cardiac events, 27 women and 28 men, aged 65–70 years. The total RNA including miRNA fraction was isolated using QiagenmiRNEasy Serum/Plasma kit according to the manufacturer’s protocol. The isolated miRNAs were analyzed using a quantitative polymerase chain reaction (qRT-PCR) technique. The plasma renalase levels were measured using a commercial ELISA kit. Results In the group of patients with high levels of renalase, higher miRNA-146a expression was found, compared with those with low concentration of renalase. Patients with simultaneous low miRNA-146a expression and high level of renalase were confirmed to deliver a significantly longer survival time compared with other patients. Conclusions miRNA-146a and plasma renalase levels were estimated as independent prognostic factors of hemodialyzed patients’ survival time. Patients with low miRNA-146a expression demonstrated a significantly longer survival time in contrast to the patients with a high expression level of miRNA-146a. Moreover, a significantly longer survival time was found in patients with high renalase activity compared with patients with low activity of the enzyme.

Plasma and erythrocyte relationship of catecholamines in hemodialysis patients.

The function of the autonomic nervous system is based on reciprocal interaction between the sympathetic and parasympathetic parts, most frequently in the form of antagonistic action on target organs. The main mediators of the sympathetic nervous system in the effectors part are catecholamines (CA), which are involved in various physiological processes. Moreover, CA also has a profound effect on the kidneys, being factors that impact on renal haemodynamics, and have been reported to be altered in pathological disorders, e.g. extracellular volume expression, hypertension and cardiovascular complications. The increased sympathetic nerve activity, at least in part, can explain the raised in plasma CA observed in chronic kidney diseases. Furthermore, plasma CA levels in ureamic patients cannot be considered a reliable index of sympathetic activity, due to existence of many factors which may affect their values. In addition, CA released into the circulation, as one of many substances, may penetrate across the cellular membranes of erytrocytes (RBC). Taking these observations together, the aim of the presented study was to investigate for the first time the plasma and erythrocyte relationship of catecholamines in haemodialysis. The studies were performed among 37 haemodialysed patients who were inhabitants of the Lublin commune. Plasma and intracellular concentration of CA were measured prior to and following haemodialysis by high performance liquid chromatography with electrochemical detection. The results suggest that RBC are able to accumulate CA at the stage of terminal renal failure; in addition, the levels of adrenaline and dopamine in RBC depend on the accumulation of urea in plasma. It was also found that the dynamic changes in concentration of RBC adrenaline are an independent predictor of mortality in haemodialysis patients.

Levels of renalase and advanced oxidation protein products with regard to catecholamines in haemodialysed patients

INTRODUCTION The main mediators of the sympathetic nervous system in the effectors part are catecholamines (CA). An increased sympathetic nerve activity observed in chronic kidney disease (CKD), is due to a raised level of CA in plasma. Renalase is a protein secreted by the kidneys, composed of 342 amino acids, which is able to metabolize the circulating CA and possibly play an important role in the regulation of sympathetic tone and blood pressure. Also, oxidative stress, defined as a disruption of the equilibrium between the generation of oxidants, is a crucial factor in the development of the inflammatory syndrome associated with CKD. The advanced oxidation protein products (AOPP) represent exquisite markers of phagocyte-derived oxidative stress. OBJECTIVE The aim of the study was to investigate the concentration of renalase and explore the associations between AOPP with regards to CA in haemodialysis (HD) patients. MATERIAL AND METHODS The study was conducted among 50 residents of the municipality and neighbouring villages in the province of Lublin, central-eastern Poland. RESULTS In the studied patients, it was found that an average concentration of renalase was 44.8 ± 6.5 μg/mL, whereas of AOPP plasma levels - 57.5 ± 21.5 μmol/L. The results demonstrated the correlation between levels of renalase and AOPP in the HD patients. Indeed, elevated levels of renalase and AOPP in HD may be due to the presence of uremic toxins in blood. The concentration of urea affects the plasma concentrations of AOPP and renalase causing a direct relationship between renalase and AOPP. However, there is no clear relationship between renalase and circulating catecholamines in HD patients.

Plasma levels of catecholamines and asymmetric dimethylarginine levels as predictive values of mortality among hemodialysis patients

Abstract The aim of the study was to assess plasma concentration of catecholamines and asymmetric dimethyl arginine levels and a possible relationship to predict the mortality rates among hemodialysis patients. The study population comprised 27 subjects, aged 65-70 years. Each patient underwent dialysis thrice a week. Furthermore, the median duration of hemodialysis was 3.5 years. Based on the conducted research, it can be concluded that the concentrations of adrenaline and the level of asymmetric dimethylarginine have predictive value of mortality among hemodialysis patients. Of note, lowering plasma asymmetric dimethylarginine concentration may represent therapeutic target for prevention of progressive renal damage.

Investigation of relationship between precursor of miRNA-944 and its mature form in lung squamous-cell carcinoma - the diagnostic value

INTRODUCTION MicroRNA (miRNA) are attractive markers of lung cancer, due to their regulatory role in cell cycle. However, we know more about function of miRNA in cancer development, there is still little known about role of their precursors (primary miRNA; pri-miRNA) in tumorgenesis. In present study we investigated potential role of miRNA-944 and its precursor pri-miRNA-944 in development of squamous-cell lung cancer (SCC) and explored interdependence between miRNA precursor and its mature form. This is a first available literature report analyzing pri-miRNA as a cancer diagnostic marker. MATERIAL AND METHODS Expression of miRNA-944 and its precursor was analyzed in 58 fresh-frozen tissues of non-small cell lung cancer and corresponding adjacent non-cancerous tissues using qRT-PCR. Expression of pri-miRNA-944 was correlated with TP63 and miRNA-944. Using ROC analysis diagnostic accuracy of studied markers was evaluated. RESULTS miRNA-944 and its precursor were significantly overexspressed in SCC compared to adenocarcinoma (AC) and non-cancerous tissue. pri-miRNA-944 strongly and positively correlated with TP63 (r = 0.739, p < 0.001) and with mature miRNA-944 expression (r = 0.691, p < 0.001). Also, TP63 expression significantly correlated with mature miRNA (r = 0.785, p < 0.001). Combined analysis of pri-miRNA-944 and mature miRNA-944 allowed to distinguish SCC tissue form AC with sensitivity of 93.3% and specificity of 100% (AUC = 0.978), and SCC from non-cancerous tissue with 92.9% sensitivity and 100% specificity (AUC = 0.992). CONCLUSION We assumed that pri-miRNA-944 and miRNA-944 may be involved in early squamous-type differentiation of lung tumors. Moreover, analysis of both markers provided high diagnostic accuracy for SCC detection.

Can chronic heart failure induce kidney function damage

Abstract Accelerated atherosclerosis and increased cardiovascular events have been extensively documented in patients with end stage chronic kidney disease. The aim of our work was to find evidence supporting the theory that chronic heart failure (CHF) induces renal function damage. In our work, lipids, apolipoprotein (apo)AI, NTproBNP, hsCRP, lipid hydroperoxide (LPO) and creatinine levels were determined in patients with CHF. A total of 37 patients who were diagnosed with CHF, as well as 15 healthy persons, were recruited for the study. The patients were placed into 2 groups: patients with NYHA class 2 and NYHA class 3. Using routine laboratory methods, NT-proBNP level, and lipids were measured by way of employing a Cobas Integra analyser, while the concentration of hs-CRP was measured by immunonephelometric methods. Moreover, serum LPO concentration was measured using Cayman’s Assay Kit (LPO). The statistical analysis of the obtained results was performed using the nonparametric Kruskal-Wallis test and Spearman’s correlation analysis. Our work demonstrated that the CHF patients had significantly decrease concentration of HDL cholesterol and apoAI, but increased NT-pro-BNP, hsCRP and LPO levels. In all CHF patients, a significant positive correlation between NT-proBNP concentration and creatinine levels, and a significant negative correlation between NT-proBNP concentration and apoAI levels, as well as between concentration of creatinine and apoAI levels, was shown. The study results suggest that variation in the concentration of NT-proBNP, LPO, hsCRP, apoAI, creatinine, in addition to chronic heart failure progression, gradually accompany the progress of chronic renal failure. What is more, the disorders may lead to the occurrence of cardiovascular events, consequently, to patient death.