[Transthyretin amyloidosis in a cohort of old and very old patients with chronic heart failure].

BACKGROUND: Life-time diagnostics of wild type transthyretin amyloidosis (ATTR(wt)-amyloidosis) is virtually absent, even though ATTR(wt)-amyloidosis is an underestimated cause for morbidity and mortality, particularly in the older age group. AIM: To study incidence, demographic characteristics, and morpho-functional features of ATTR(wt)-amyloidosis in patients with FC IV CHF and LV hypertrophy > 15 mm according to autopsy data. MATERIALS AND METHODS: Postmortem reports were retrospectively analyzed for patients (n=141; 19 % males, 81 % females) of cardiology departments aged ≥69 with the underlying CHF syndrome. From all formalin-fixed fragments of the myocardium embedded in paraffin were prepared 5-7 mkm cuts, which were stained with Congo red (Sigma, USA) and viewed under normal and polarized light. Immunohistochemical analysis was also performed using antibodies to AA-amyloid, transthyretin, kappa and lambda-light chains of immunoglobulins. RESULTS: deposits were found in both old and very old persons aged 91.25±9.67, mostly in women due to shorter life span of men. In different FCs associated with LV hypertrophy, according to autopsy data amyloid deposits were observed in virtually every fifth deceased (21 % of cases). The amount of myocardial amyloid deposits was generally small (56 % of cases had (+) and 27 % had (++) amyloid deposits); 17 % of cases had considerable amyloid deposits (7 % had (+++) and 10 % had (++++)). The presence of amyloid deposits did not influence indexes of myocardial hypertrophy, such as ventricular septum thickness, LV posterior wall thickness, and heart mass. In the presented cases we observed focal amyloid deposition in the myocardium typical for old age-related amyloidosis; in 97 % cases, amyloid was located in the interstitium, around cardiomyocytes and in 3 % of cases - exclusively around blood vessels. CONCLUSION: ATTR (wt)-amyloidosis was detected in every fifth patient in the old and very old cohort, primarily in women (83 %), and was not diagnosed during the life time. Characteristic morphological manifestations of ATTR(wt)-amyloidosis were focal amyloid deposits mostly in the myocardial interstitium.

CHARACTERISTICS OF AMYLOID DEPOSITS DETECTED IN THE INTERNAL ORGANS OF mdx MICE.

The dystrophin-deficient mdx mouse is the most commonly used experimental model of Duchenne muscular dystrophy (DMD). Although the amyloid has been shown in the muscle biopsies of patients with different types of muscular dystrophies, there are no data on the amyloid accumulations in the biopsy of DMD patients or mdx mouse. Therefore, the aim of the present study was to testify the hypothesis of probable accumulation of amyloid in the visceral organs of mdx mouse. Specimens of myocardium, kidneys, and liver of male and female mdx mice aged from 2 months to 1.5 years (n = 9) were used in the study. The histochemical staining with Congo red demonstrated amyloid accumulations in the studied organs of the mdx mice. Morphology and localization of the found accumulations were described in details and analyzed. The mass-spectrometric study determined the vitronectin and apolipoprotein A-II as the most probable components of the amyloid accumulations in the mdx mouse.

[STEM CELLS PLAY NO CONSIDERABLE ROLE IN CARDIOMYOCYTE REPOPULATION OF ADULT HUMAN HEART].

There are two viewpoints concerning cardiac regeneration. One assumes that the myocardium of an adult human heart has a weak regenerative capacity. According to another, myocardium can renew at a high rate due to the presence of resident stem cells. This study was aimed to test the role of stem cells in myocardium repopulation in adult humans of different age by examining the distribution of cardiomyocytes as to their size and ploidy. Cytofluorimetry and interferometry were used to determine the dry weight, volume and ploidy of myocytes isolated from the left ventricle of the normal heart of 12 men aged 20-30 years (n = 7) and 40-50 years (n = 5). Dry weight of cardiomyocytes made up 6906 ± 182 pg (10(-12) g) aged 20-30 years and 9126 ± 263 pg in men aged 40-50 years. There were no cells with an intermediate volume between amplifying and mature myocytes. The number of candiomyocytes in the left ventricle made up (3.18 ± 0.05) x 10(9) cells in the age group 20-30 years and (2.06 ± 0.6) x 10(9) cells in the age group 40-50 years. Most of the myocyte population was represented by mononucleate cells with tetraploid nuclei (41.3%). Proportion of myocytes of different ploidy classes did not change in the interval from 20 to 50 years. Our results strongly suggest that stem cells of the heart are not involved in the regeneration of human myocardium during aging. The function of the aging heart is mostly compensated by the hypertrophy of the remaining myocytes.

[The informative value of oral mucosal biopsy for the diagnosis of systemic amyloidosis].

One hundred and thirteen oral mucosal biopsy specimens were analyzed in patients suspected of having systemic amyloidosis. Histological, immunohistochemical, and genetic studies and polarized light microscopy revealed oral amyloid deposits in 72.6% of cases, including 63.5% with metabolic syndrome and 36.5% with another etiology of chronic heart failure (coronary heart disease, cardiomyopathy). Systemic amyloidosis was found in 13.4% of cases (hereditary transthyretin, AL, and senile forms in 1.8, 10.5, and 1.1% cases, respectively). An anterior abdominal wall skin flap was a valid location to reveal the systemic forms of amyloidosis. Patients with metabolic syndrome and periodontal diseases may have local oral amyloidosis.

[Oral status and oral mucosa blood circulation changes in patients with chronic heart failure].

The objective of this study was to characterize dental status and oral mucosa blood flow in patients with chronic heart failure and amyloid deposits in oral mucosa. Histological and immunohistochemical analysis of 80 oral mucosa biopsies taken from patients aged 32-72 years with chronic heart failure I-IV NYHA functional class was carried out. It detected a systemic amyloidosis in 15.7% of cases; a local amyloid deposition in oral mucosa was found in 58.5% of cases. Amyloid deposition in oral mucosa was associated with severe chronic generalized periodontitis in more than a half of cases. Amyloid deposits in oral mucosa were revealed more often in patients with metabolic syndrome (63.5%). The article describes dental status and oral mucosa blood flow in patients with heart failure.

[Transthyretin gene V30M, H90N, and del9 mutations in cardiomyopathy patients from St. Petersburg].

A search of transthyretin (TTP) gene mutations was conducted in patients with cardiomyopathies from St. Petersburg. Mutations H90N, V30M, G47A, and deletion (del9) of nucleotides GACTTCTCC in position 6776 from the start codon of the TTP gene (in position 98782 according to reference sequence AC079096 (NCBI) was found. The H90N mutation in the third exon of TTP gene was detected in a son of a cardiomyopathy patient and in his mother, which lacked any clinical manifestations. Mutations V30M and G47A in exon 2 of TTP gene were found in heterozygous and homozygous state, respectively, in one of the probands. Deletion (del9) was revealed in a patient with cardiomyopathy and in his two daughters from different marriages, who had no clinical manifestations of the disease. All the mutations revealed in this study were previously identified in other populations.

[Desmin-related cardiomyopathy].

The observation of 26 years old patient with desminopathy declared itself by hypertrophied cardiomyopathy with its transformation into restrictive phenotype is presented. The features of pathologic course at the patient were a dominance and diversity of cardiac manifestations. Endomyocardiac biopsy allowed suspecting the desminopathy confirmed by genetic analysis. Morphological features of desmin-related cardiomyopathy were irregular desmin conglomerates mainly located under sarcolemma and an indirect histological signs of idiopathic cardiomyopathy as well nuclear polymorphism, perinuclear "nimbus", chaotic located myofibrils.

[Cardiac manifestations of amyloidosis].

Amyloidosis is shown by a group of diseases with a variability of clinical and morphological manifestations determining difficulties in its diagnosis. The cardiovascular system is commonly the site of involvement in different forms of amyloidosis. The variants of cardiac involvement in the pathological process in amyloidosis are outlined depending on various precursor proteins. The specific features of different forms of cardiopathic amyloidosis are described in relation to its type.

[Cellular aspects of hypertrophic cardiomyopathy pathogenesis: the role of cardiomyocyte polyploidy and activation of the proliferating cell nuclear antigen in the myocardium].

Mechanisms of hypertrophy development in hypertrophic obstructive cardiomyopathy (HOCM) have not been enough investigated. In our study, there have been examined patients with severe HOCM at different ages, including children, and patients with essential arterial hypertension (EAH). There was found, that HOCM in children compared to adults was characterized by considerable interventricular septum (IVS) hypertrophy and it was accompanied by the acceleration of cardiomyocyte polyploidy. The average ploidy level of cardiomyocytes in children with HOCM was higher than analogous indices in adults. The average ploidy level of nuclei, the part of PCNA-positive nuclei and polyploidic nuclei of cardiomyocytes in aduls with HOCM were authentically higher than in patients with EAH. Activation of the nuclear antigen in stromal cells was detected only in patients with HOCM. Our findings provide evidence of an important role of cardiomyocyte polyploidy and activation of the proliferating cell nuclear antigen in development of the myocardial hypertrophy in patients with HOCM.

[Elevated expression of argentophilic proteins from the nucleolar organizer regions in myocardium of patients with obstructive hypertrophic cardiomyopathy and mutations in p53 tumor suppressor gene]. / Povyshennaia ékspressiia argentofil'nykh belkov oblastei iadryshkovogo organizatora v miokarde bol'nykh obstruktivnoi gipertroficheskoi kardiomiopatiei i mutatsii v gene opukholevogo supressora p53.

Silver staining for nucleolar proteins was used to evaluate the ribosomal genes activity in cardiomyocytes (Cm) and fibroblast-like cells (FBS) of intraventricular septum, and other regions of the left ventricle. Specimens to be analysed were taken from 7 patients with idiopathic obstructive hypertrophic cardiomyopathy (OHCM). In this group the quantity of nucleoli and AgNORs, reflecting the transcriptional and processing level of pre-ribosomal RNA in all type of cells, was at least 2 and 3 times higher than in patients with essential hypertension and in healthy control, respectively. We suggest that nucleolar hypertrophy in patients with hypertrophic cardiomyopathies, inappropriate to hypertrophy in other pathological conditions, may be most probably of compensatory character, and this may be in part explained by nuclear hyperploidy and chromosomal endoreduplication. We have noted a marked heterogeneity in shape, size and quantity of nucleoli, and in AgNORs of FBC. Nuclei with modulated phenotype containing nucleoli of high activity were revealed. This article presents the first data on p53 mutation identification in patients with advanced OHCM.

[Mesenchymal stem cell transplantation for myocardial reparation of rat experimental heart failure]. / Terapiia éksterimental'nogo infarkta miokarda u krys s pomoshch'iu transplantatsii singennykh mezenkhimnykh stvolovykh kletok.

Mesenchymal stem cells (MSC) are resident pluripotent cells of bone marrow stroma. MSC have the ability to differentiate into osteoblasts, chondroblasts and adipocytes, neurons, glia and also into cardiomyocytes. The problem of MSC use in cell therapy of various diseases and in myocardial infarction therapy is widely discussed at present. The experiments were carried out on the inbred line Wistar--Kyoto rats. Myocardial experimental infarction (EI) was induced by left descending coronary artery ligation. MSC were isolated from bone marrow, cultivated in vitro and injected into the tail vein on the day of experimental infarction operation. It was shown that the structure of injured myocardium in experimental group significantly differed from that in control group. MSC transplantation led to inflammatory process acceleration and to increased angiogenesis in the damaged myocardium; also, live cardiomyocyte layers were detected in the scar. As a result, ventricular dilatation and overload of the border zone of infarct region decreased, no features of infarction relapse were shown in the border zone.