Genomic and Transcriptomic Research in the Discovery and Application of Colorectal Cancer Circulating Markers.

Colorectal cancer (CRC) is the most frequently occurring malignancy in the world. However, the mortality from CRC can be reduced through early diagnostics, selection of the most effective treatment, observation of the therapy success, and the earliest possible diagnosis of recurrences. A comprehensive analysis of genetic and epigenetic factors contributing to the CRC development is needed to refine diagnostic, therapeutic, and preventive strategies and to ensure appropriate decision making in managing specific CRC cases. The liquid biopsy approach utilizing circulating markers has demonstrated its good performance as a tool to detect the changes in the molecular pathways associated with various cancers. In this review, we attempted to brief the main tendencies in the development of circulating DNA and RNA-based markers in CRC such as cancer-associated DNA mutations, DNA methylation changes, and non-coding RNA expression shifts. Attention is devoted to the existing circulating nucleic acid-based CRC markers, the possibility of their application in clinical practice today, and their future improvement. Approaches to the discovery and verification of new markers are described, and the existing problems and potential solutions for them are highlighted.

Ventricular tachycardia incidence and erythrocyte membranes ß-adrenoreactivity in patients with implanted cardioverter-defibrillator.

BACKGROUND: In patients with high risk of ventricular tachycardia (VT) the proven beneficial therapy is the implantable cardioverter defibrillator (ICD). It has been shown that the coronary artery disease (CAD) and VT development are accompanied by a persistent change of the sympathoadrenal system activity. This leads to a decrease in the total density of the erythrocyte membrane ß-adrenergic receptors. The purpose of this study was to identify the relationship of the erythrocyte membranes ß-adrenoreactivity (EMA) with VT development in patients with CAD and ICD. METHOD: Sixty-three patients (male - 53, age - 66.6 ± 9.2 years) with CAD and ICD were included to the study. EMA was studied using a method for assessing erythrocyte osmoresistance increase as a result of ß-adrenergic receptors blockade by a selective ß-adrenergic blocker. VT and ventricular fibrillation (VF) events recorded by ICD were evaluated. RESULTS: The 1st group consist of 23 patients with VT/VF events recorded by ICD during 27.0 [14.0; 53.0] months follow-up period. EMA indicator in this group was 41.54% [27.15; 51.26]. The 2nd group consist of 40 patients without VT/VF events and the same indicator was significantly higher - 55.42% [35.67; 62.33] (p = .04). The ROC-analysis (AUC = 0.657; Sen = 78.26; Spe = 55.00; p = .031) and binary logistic regression (OR = 0.9679; 95% CI: 0.9384-0.9983; p = .038) showed that EMA indicator 51.26% or lower was the independent predictor of VT events. CONCLUSIONS: In patients with CAD and ICD erythrocyte membranes ß-adrenoreactivity indicator 51.26% or lower is the predictor of VT episodes.

Nonpharmacological Correction of Hypersympatheticotonia in Patients with Chronic Coronary Insufficiency and Severe Left Ventricular Dysfunction.

BACKGROUND: Control of sympathetic hyperactivity is pivotal for treatment of heart failure (HF) in patients with coronary artery disease (CAD). Our earlier studies demonstrated that the auricular pulsed electrical stimulation of the vagus nerve (VNS) beneficially affected condition of CAD patients with HF. The aim of our study was to evaluate changes in heart rate (HR) and the levels of heat shock proteins in peripheral blood lymphocytes in patients with CAD in the course of VNS. METHODS: The study comprised 70 individuals aged 50-68 years with chronic coronary insufficiency, severe left ventricular dysfunction, and NYHA functional class (FC) III-IV HF. Main group included 63 patients who received VNS course (group 1). Control patients (n = 7) received sham therapy (group 2). RESULTS: According to the results of 6-minute walk test and 24-hour ECG monitoring, administration of VNS improved clinical condition of 58 of 63 patients, decreased HF FC, and attenuated HR. Clinical condition in sham therapy group did not change. Immunoenzyme method demonstrated that hsp70 and hsp60 contents in peripheral blood lymphocyte lysate increased by 58% and 48% (P < 0.05), respectively, in patients who initially had HR < 80 bpm. The hsp70 level significantly increased and hsp60 level remained unchanged in patients with initial HR > 80 bpm. CONCLUSIONS: Correction of autonomous nervous status by VNS attenuated HR and improved functional state of the heart in CAD patients. Cardiotropic effect of VNS was the most pronounced in patients with preserved endogenous stress-limiting systems associated with hsp60 and/or hsp70.

Dynamic Changes in Circulating Methylated Markers in Response to Antitumor Therapy of Rectal Cancer.

BACKGROUND: Rectal cancer (RC) occupies a leading position in the structure of oncological morbidity and mortality. Aberrant methylation of tumor-suppressor genes and hypomethylation of retrotransposons were shown to be detectable in cell-free DNA, circulating in the blood (cfDNA) of cancer patients, indicating the possibility to use them as diagnostic and prognosis markers. PURPOSE: Evaluation of the changes in the methylation level of LINE-1 elements and SEPTIN9 and IKZF1 genes in the cell-surface-bound cfDNA (csb-cfDNA) from the blood of RC patients after antitumor therapy at a long-term follow-up. METHODS: Blood samples were obtained from RC patients (n = 25) before treatment, after preoperative chemotherapy (3 courses according to the XELOX scheme), 10-15 days after surgery, and every 3 months during 12 months of dynamic observation. The methylation level of LINE-1, SEPTIN9, and IKZF1 in the csb-cfDNA was evaluated by quantitative methyl-specific PCR. RESULTS: The LINE-1 methylation level in the csb-cfDNA increased 1.6 times in RC patients after chemotherapy and 3 times after tumor resection versus methylation level before therapy. The SEPTIN9 gene methylation level in the csb-cfDNA decreased by 1.7 times in RC patients after chemotherapy and by 2.3 times after tumor resection compared with the values before the treatment. The IKZF1 gene methylation level decreased by 2 times in RC patients after combined therapy. Notably, all patients with relapses (n = 5) showed an increase in methylation level for the SEPTIN9 and IKZF1 genes and a decrease of methylation level for the LINE-1 elements by 2 times or more in comparison with the level 10-15 days after surgery. There were no changes in the circulating SEPTIN9, IKZF1, and LINE-1 methylation levels during the 12-month follow-up period after the combined therapy of RC patients (n = 20) without relapses. CONCLUSION: The results indicate that SEPTIN9, IKZF1, and LINE-1 methylation levels in the csb-cfDNA are potential markers of the effectiveness of antitumor therapy and early detection of relapse in RC patients.

Predictors of appropriate therapies delivered by the implantable cardioverter-defibrillator in patients with coronary artery disease during long-term period.

This prospective study aimed to investigate the ability of cardiac autonomic nervous system (CANS) activity assessment to predict appropriate implantable cardioverter-defibrillator (ICD) therapy in patients with coronary artery disease (CAD) during long-term follow-up period. We enrolled patients with CAD and ICD implantation indications that included both secondary and primary prevention of sudden cardiac death. Before ICD implantation CANS was assessed by using heart rate variability (HRV), myocardium scintigraphy with 123I-meta-iodobenzylguanidine (123I-MIBG) and erythrocyte membranes ß-adrenoreactivity (EMA). The study's primary endpoint was the documentation of appropriate ICD therapy. Of 45 (100.0%) patients, 15 (33.3%) had appropriate ICD therapy during 36 months follow-up period. Patients with appropriate ICD therapy were likely to have a higher summed 123I-MIBG score delayed (p < 0.001) and lower 123I-MIBG washout rate (p = 0.008) indicators. These parameters were independently associated with endpoint in univariable and multivariable logistic regression. We created a logistic equation and calculated a cut-off value. The resulting ROC curve revealed a discriminative ability with AUC of 0.933 (95% confidence interval 0.817-0.986; sensitivity 100.00%; specificity 93.33%). Combined CANS activity assessment is useful in prediction of appropriate ICD therapy in patients with CAD during long-term follow-up period after device implantation.

Comparative analysis of hypertensive nephrosclerosis in animal models of hypertension and its relevance to human pathology. Glomerulopathy.

Current research on hypertension utilizes more than fifty animal models that rely mainly on stable increases in systolic blood pressure. In experimental hypertension, grading or scoring of glomerulopathy in the majority of studies is based on a wide range of opinion-based histological changes that do not necessarily comply with lesional descriptors for glomerular injury that are well-established in clinical pathology. Here, we provide a critical appraisal of experimental hypertensive glomerulopathy with the same approach used to assess hypertensive glomerulopathy in humans. Four hypertensive models with varying pathogenesis were analyzed-chronic angiotensin II infused mice, mice expressing active human renin in the liver (TTRhRen), spontaneously hypertensive rats (SHR), and Goldblatt two-kidney one-clip rats (2K1C). Analysis of glomerulopathy utilized the same criteria applied in humans-hyalinosis, focal segmental glomerulosclerosis (FSGS), ischemic, hypertrophic and solidified glomeruli, or global glomerulosclerosis (GGS). Data from animal models were compared to human reference values. Kidneys in TTRhRen mice, SHR and the nonclipped kidneys in 2K1C rats had no sign of hyalinosis, FSGS or GGS. Glomerulopathy in these groups was limited to variations in mesangial and capillary compartment volumes, with mild increases in collagen deposition. Histopathology in angiotensin II infused mice corresponded to mesangioproliferative glomerulonephritis, but not hypertensive glomerulosclerosis. The number of nephrons was significantly reduced in TTRhRen mice and SHR, but did not correlate with severity of glomerulopathy. The most substantial human-like glomerulosclerotic lesions, including FSGS, ischemic obsolescent glomeruli and GGS, were found in the clipped kidneys of 2K1C rats. The comparison of affected kidneys to healthy control in animals produces lesion values that are numerically impressive but correspond to mild damage if compared to humans. Animal studies should be standardized by employing the criteria and classifications established in human pathology to make experimental and human data fully comparable for comprehensive analysis and model improvements.

A novel method for comparison of arterial remodeling in hypertension: Quantification of arterial trees and recognition of remodeling patterns on histological sections.

Remodeling of spatially heterogeneous arterial trees is routinely quantified on tissue sections by averaging linear dimensions, with lack of comparison between different organs and models. The impact of experimental models or hypertension treatment modalities on organ-specific vascular remodeling remains undefined. A wide variety of arterial remodeling types has been demonstrated for hypertensive models, which include differences across organs. The purpose of this study was to reassess methods for measurement of arterial remodeling and to establish a morphometric algorithm for standard and comparable quantification of vascular remodeling in hypertension in different vascular beds. We performed a novel and comprehensive morphometric analysis of terminal arteries in the brain, heart, lung, liver, kidney, spleen, stomach, intestine, skin, skeletal muscle, and adrenal glands of control and Goldblatt hypertensive rats on routinely processed tissue sections. Mean dimensions were highly variable but grouping them into sequential 5 µm intervals permitted creation of reliable linear regression equations and complex profiles. Averaged arterial dimensions demonstrated seven remodeling patterns that were distinct from conventional inward-outward and hypertrophic-eutrophic definitions. Numerical modeling predicted at least nineteen variants of arterial spatial conformations. Recognition of remodeling variants was not possible using averaged dimensions, their ratios, or the remodeling and growth indices. To distinguish remodeling patterns, a three-dimensional modeling was established and tested. The proposed algorithm permits quantitative analysis of arterial remodeling in different organs and may be applicable for comparative studies between animal hypertensive models and human hypertension. Arterial wall tapering is the most important factor to consider in arterial morphometry, while perfusion fixation with vessel relaxation is not necessary. Terminal arteries in organs undergo the same remodeling pattern in Goldblatt rats, except for organs with hemodynamics affected by the arterial clip. The existing remodeling nomenclature should be replaced by a numerical classification applicable to any type of arterial remodeling.

Influence of Amiodarone and Dronedarone on the Force-Interval Dependence of Rat Myocardium.

The antiarrhythmic effect of amiodarone and its analogue dronedarone is caused by their direct actions on several cardiomyocyte sarcolemmal ion currents. However, whether their effects are related to intracellular calcium levels is not exactly known. Ca2+ cycling refers to the release and reuptake of intracellular Ca2+, which induces muscle contraction and relaxation and determines the force-interval dependence. This study aimed to evaluate the influence of amiodarone and dronedarone on the force-interval relationship. Materials and Results. The work was performed on the papillary muscles of the left ventricle of male Wistar rats. Muscle perfusion was performed at 36.5°C with oxygenated Krebs-Henseleit solution with baseline stimulation 0.5 Hz. The postrest test (4-60 s) and the extrasystolic exposure (0.2-1.5 s) were evaluated. Inotropic reaction to the test exposure was evaluated before and after muscle perfusion with solution containing amiodarone (10-6 M) or dronedarone (10-6 M) during 10 min. Amiodarone or dronedarone led to decrease of the amplitude of extrasystolic contractions of the papillary muscles. The amplitude of postextrasystolic contractions after short extrasystolic intervals on the background of the drugs was increased. Amiodarone and dronedarone led to increase of the amplitude of postrest contractions. Conclusions. Dronedarone reduces the excitability of cardiomyocyte sarcolemma to a greater extent than amiodarone. Amiodarone and dronedarone are able to increase postextrasystolic and postrest potentiation. The effect of amiodarone on postextrasystolic and postrest potentiation is more pronounced in comparison with dronedarone.

Comparative analysis of changes of myocardial angiogenesis and energy metabolism in postinfarction and diabetic damage of rat heart.

Comparative study of changes in myocardial activity of lactate dehydrogenase (LDH), succinate dehydrogenase (SDH), and capillary density distribution in the experimental models of diabetic and postinfarction damage of rat heart was performed. Data showed that decrease in LDH and SDH activities was observed in both pathologies which can suggest abnormal processes of glycolysis and oxidative phosphorylation in cardiac mitochondria. Activity of LDH and SDH in combined pathologies was comparative with the corresponding values of these parameters in control group. The authors hypothesize that these differences can be caused by specifics of myocardial vascularization. The results of the study showed that an increase in capillary density was found in all groups of rats with pathologies compared with control group. However, no significant differences in the intensity of angiogenesis processes were found between groups with pathologies.