Inflammation and oxidative stress caused by nitric oxide synthase uncoupling might lead to left ventricular diastolic and systolic dysfunction in patients with hypertension

Objective: To investigate the role of oxidative stress, inflammation, hypercoagulability and neuroendocrine activation in the transition of hypertensive heart disease to heart failure with preserved ejection fraction (HFPEF). Methods: We performed echocardiography for 112 patients (≥ 60 years old) with normal EF (18 controls and 94 with hypertension), and determined protein carbonylation (PC), and tetrahydrobiopterin (BH4), C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), fibrinogen, plasminogen activator inhibitor type-I (PAI-I), von Willebrand factor, chromogranin A (cGA) and B-type natriuretic peptide (BNP) levels from their blood samples. Results: We found that 40% (38/94) of the patients with hypertension (HT) had no diastolic dysfunction (HTDD-), and 60% (56/94) had diastolic dysfunction (HTDD+). Compared to the controls, both patient groups had increased PC and BH4, TNF-α, PAI-I and BNP levels, while the HTDD+ group had elevated cGA and CRP levels. Decreased atrial and longitudinal left ventricular (LV) systolic and diastolic myocardial deformation (strain and strain rate) was demonstrated in both patient groups versus the control. Patients whose LV diastolic function deteriorated during the follow-up had elevated PC and IL-6 level compared to their own baseline values, and to the respective values of patients whose LV diastolic function remained unchanged. Oxidative stress, inflammation, BNP and PAI-I levels inversely correlated with LV systolic, diastolic and atrial function. Conclusions: In patients with HT and normal EF, the most common HFPEF precursor condition, oxidative stress and inflammation may be responsible for LV systolic, diastolic and atrial dysfunction, which are important determinants of the transition of HT to HFPEF

(Int J Mol Sciences)

Liquid biopsy has recently become a very popular, noninvasive sampling procedure. The mainadvantage of this method is the possibility of easy cell-free nucleic acids isolation from differentfluids, primarily blood, and their use in diagnostic or screening processes. The other advantage isthe possibility to follow up on patients’ treatments. Liquid biopsies are applicable in the diagnosis ofoncological, cardiovascular, and infectious diseases and in the use of noninvasive prenatal testing(NIPT). Gene expression studies on gDNA, mtDNA, miRNA, and lncRNA can provide valuableinformation to researchers and clinicians. Currently, it is very popular to obtain exosomes from liquidbiopsies and use them in similar studies. Understanding their role in different pathological conditionscould provide us with valuable information on cell–cell-tissue communication. The identification ofnew extra- and intra-cellular signaling of nucleic acid molecules and pathways could help in thediagnosis and treatment of patients. Cell-free DNAs are widely used in the prenatal detection ofgenetic diseases. Recently oncological diseases have been a focus of research and the newest area tobe researched is cardiovascular disease.L

Noninvasive prenatal testing for congenital heart disease – cell-free nucleic acid and protein biomarkers in maternal blood

Context: Congenital heart disease (CHD) is the most common fetal malformation. Prenatalultrasonography is routinely applied for the screening of CHD but many factors influence itsdiagnostic accuracy. The introduction of new biomarkers could facilitate the identification ofhigh-risk pregnancies.Objective: In our review, our aim was to collect expression studies of cell-free nucleic acidsand proteins in maternal circulation. Syndromic CHDs which can be detected by non-invasiveprenatal testing (NIPT) techniques were also discussed.Methods:PubMed and Web of Science databases were screened for studies where the levelsof potential CHD biomarkers were measured in maternal blood samples. Available NIPT testswere collected from the providers’ resources.Results:There are nine CHD-associated chromosomal abnormalities, five aneuploidies, andfour microdeletions, which are included in NIPT panels.We found eight articles from which fiveincluded the analysis of specific cell-free RNA expression and three measurements of proteinlevels.Conclusion: Most of the common heart-related chromosomal aberrations can be diagnosed byNIPT. Specific cell-free RNAs and circulating proteins seem to be potential biomarkers forfetal CHDs. The application of these new biomarkers could improve the detection rate at earlypregnancy, making it possible to provide optimal perinatal and perioperative management

Overexpression of CD24, c-myc and Phospholipase 2A in Prostate Cancer Tissue Samples Obtained by Needle Biopsy

Altered CD24, c-myc and phospholipase 2a expression was reported in different cancers. Our aim was to measure the expression of these genes in prostate cancer tissues, and compare it to non-cancerous samples. Prostate tissue samples were collected by needle biopsy from 20 prostate cancer (PCA) and 11 benign prostate hyperplasic (BPH) patients. RNA was isolated; cDNA synthetized, CD24, c-myc and phospholipase 2A (PL2A) expressions were determined by quantitative real-time PCR method. The expression of beta-globin gene was measured for normalization of the gene expression results. Serum prostate specific antigen (PSA) levels were determined by microparticle enzyme immunoassay (MEIA) method. PSA levels were significantly different between the PCA and BPH groups, 252.37 +/- 308.33 ng/ml vs. 3.5 +/- 2.14 ng/ml (p = 0.001), respectively. CD24 expression was 988.86 +/- 3041 ng/microl in prostate tumor and 4.00 +/- 4.25 ng/microl in the BPH group (p = 0.035). The c-myc expression was 88.32 +/- 11.93 ng/microl in the prostate tumor and 17.08 +/- 21.75 ng/microl in the BPH group (p = 0.02), and the PL2A 31.36 +/- 67.02 ng/microl was in PCA and 5.56 +/- 14.08 ng/microl in BPH (p = 0.025). Gleason's scores showed correlation with c-myc (p = 0.019) and PSA (p = 0.033) levels. Overexpression of PL2A, CD24 and c-myc was observed in prostate cancer samples using quantitative real-time PCR method

Increased B-type natriuretic peptide levels in early-onset versus late-onset preeclampsia

Background: We compared B-type natriuretic peptide (BNP) levels, clinical and laboratory findings in early-onset preeclampsia (EOP), late-onset preeclampsia (LOP) and healthy pregnant groups.Methods: We studied 40 healthy pregnant and 40 preeclamptic patients. Preeclamptics were divided in two groups, the EOP group (n=20) and LOP group (n=20), according to gestational age at the onset of disease. The distinction criterion for early- vs. late-onset was set as week 34 of gestation. The concentration of the BNP levels was measured by a sandwich fluorescence immunoassay. For statistical analysis of the clinical and laboratory findings non-parametric methods were applied.Results: BNP levels were higher in EOP [61.35 (36.95–93.25) pg/mL] and LOP patients [32.4 (19.15–39.2) pg/mL] than in healthy pregnant women [10.05 (6.08–16.03) pg/mL] (both p<0.001). Furthermore, EOPs had significantly higher BNP levels as compared to LOP patients (p<0.001). A BNP cut-off <24.5 pg/mL had a negative-predictive value of 85.1% excluding preeclampsia. There was a significant inverse correlation between plasma BNP levels of EOP patients and sodium (p<0.05) and total protein concentrations (p<0.05). In the EOP group, a significant positive correlation was observed between plasma levels of BNP and hematocrit (p<0.05), serum potassium (p<0.05), urea (p<0.05) and 24-h proteinuria (p<0.05).Conclusions: BNP levels were significantly higher in EOP than in LOP patients. The cut-off value <24.5 pg/mL seems to be a powerful discriminative indicator excluding preeclampsia. The amount of proteinuria and total protein levels correlate with the elevation of the BNP levels. In EOP the extent of proteinuria is higher than in the LOP

Cell-free, long non-coding RNA in the diagnosis of ovarian cancerr

Background: Ovarian cancer is the one of leading causes of cancer-associated mortality among women. An optimalised and established non-invasive diagnostic method may contribute to the early diagnosis of this malignancy. The significant role of the long non-coding RNAs (lncRNAs) was described in the oncogenesis, metastasis and chemoresistance. Upregulation of the one of lncRNA, namely, Hox transcript antisense intergenic RNA (HOTAIR) was determined in the development of ovarian cancer cells.Materials and methods: Twenty previously untreated ovarian cancer patients (60.60±10.84y; FIGO stages III or IV) and 20 healthy controls (56.70±14.51y) were studied. EDTA-anticoagulated blood was drawn; RNA was isolated; cDNA was synthesised and the HOTAIR lncRNA expression were determined by using ExiLERATE LNA™ qPCR,for mRNA and long non-coding RNA (Exiqon, USA). Beta-actin was used as reference gene. Student t-test was used for the statistical calculations.Results: Higher expression of HOTAIR was detected in the cell-free plasma samples of patients with ovarian cancer (1.89±2.39) compared to healthy controls (1.39±1.48), while the statistical analysis did not show significant difference.Conclusion: This is the first HOTAIR expression determination from the cell-free plasma of ovarian cancer patients and healthy controls, according to our knowledge. No significant difference was determined in the expression of the analysed lncRNA between Hungarian ovarian cancer patient and healthy control groups

Relationship between cardiovascular diseases and circulating cell-free nucleic acids in human plasma

Cardiovascular diseases (CVDs) are the main cause of human morbidity and mortality worldwide. Early diagnosis could improve the efficiency of treatments. New biomarkers are needed for the identification of high-risk populations in order to make accurate diagnosis and therapy monitoring. Circulating cell-free nucleic acids (cf-NAs) offer a promising new noninvasive tool. These have a role in the regulation of normal physiological functions and in the development of pathological alterations. There is extended research on the clinical application and utilization of cell-free genomic DNA, mtDNA, mRNA, miRNA and long noncoding RNA in CVDs. These molecules could serve as components of new generation therapeutics. Our review focuses on the role of cf-NAs in the pathogenesis of CVDs and we are discussing also possible diagnostic applications and therapeutic implications

Circulating exosomal and Argonaute-bound microRNAs in preeclampsia

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Circulating exosomal and Argonaute-bound microRNAs in preeclampsia

Introduction: microRNAs (miRNAs) play important role in the regulation of placental development, and abnormal miRNA expression is associated with preeclampsia (PE). miRNAs are released from trophoblast cells to maternal blood flow, where they are highly stable, being encapsulated inside extracellular vesicles, like exosomes or bound to Argonaute proteins. In PE, placental dysfunction leads to aberrant extracellular miRNA secretion. hsamiR- 210 is a hypoxia-sensitive miRNA found to be upregulated in PE, however, it is unknown whether it is the cause or the consequence of the disease. Objective: Our aim was to analyze the expression of several miRNAs, including hsa-miR- 210 in placenta, exosome and Ago-bound fractions comparing normal (N) and PE pregnancies. We performed in vitro analyses of extracellular hsa-miR-210 secretion of trophoblast cell cultures (of villous and extravillous origin) under hypoxic condition. Methods: PE and N placenta samples were collected from C-sections, and blood samples were drawn from each pregnant woman in the third trimester. Htr-8 and Jar cell lines were cultured in exosome-free media and treated with hypoxia-mimetic agents. Exosome and Agobound fractions were isolated by membrane affinity spin column method from plasma and cell media. Short RNAs were extracted from exosomes and vesicle-free fractions, and total-RNA was isolated from the placenta samples. The RNA purity and concentration were measured by spectrophotometry. Expression analysis was carried out by qPCR with specific primers to target and reference miRNAs. Results: The level of hsa-miR-210 was significantly higher in PE placentas, which could cause a minor increase of exosomal and a high elevation of Ago-bound miR-210 in circulation. Hypoxia leads to intracellular hsa-miR-210 upregulation in trophoblast cell lines. In extravillous cell (HTR8) media, only the level of exosomal hsa-miR-210 was increased but no change in Ago-bound hsa-miR-210 level was observed. In contrast, in villous cell (JAR) media, the level of exosomal hsa-miR-210 was increased and enhanced release of Ago-bound hsa-miR-210 was also observed. Conclusion: Based on our data, we postulate that in PE, exosomal hsa-miR-210 are secreted actively from the trophoblast, and by intercellular communication, it may have a role in disease etiology. In addition, there is a passive release of Ago-bound hsa-miR-210 into the circulation, which may represent by-products of cell-death and is thereby a possible consequence of the disease

Characteristic Laboratory Changes in Pregnancies Complicated by HELLP Syndrome

INTRODUCTION:HELLP syndrome (hemolysis, elevated liver enzymes, and low platelet count) is a severe, life-threatening form of preeclampsia. Its development is accompanied by significant increase in maternal, as well as fetal, morbidity, and mortality rates. It is essential, therefore, for obstetricians to be familiar with the disease.MATERIALS AND METHODS:In the past 10 years, 107 patients were treated for HELLP syndrome in the intensive care unit (ICU) of the First Department of Obstetrics and Gynaecology, Semmelweis University. During this time, we studied the characteristic laboratory findings of the disease from the day of the diagnosis until the first few postpartum days.RESULTS:HELLP syndrome was present in 0.37% of all women having live births. In our study, the liver enzymes AST, and LDH, and the level of total bilirubin (indicating the degree of hemolysis), and repeated thrombocyte counts were suitable for following the cases. The AST, LDH and bilirubin levels returned to normal between the third and seventh days postpartum. The platelet count passed the critical level of 100,000/microL on the third to fourth day.CONCLUSIONS:We have found that the platelet count, LDH, AST, and total bilirubin levels proved to be useful indicators of the progression of HELLP syndrome